\n

\n \n 2024\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n Special Issue \"Personalized Medicine in Blood Disease of Children\".\n \n \n \n\n\n \n Ceci, A., Kountouris, P., Didio, A., & Bonifazi, F.\n\n\n \n\n\n\n Journal of Personalized Medicine, 14(3): 285. March 2024.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

\n

@article{ceci_special_2024,\n\ttitle = {Special {Issue} "{Personalized} {Medicine} in {Blood} {Disease} of {Children}"},\n\tvolume = {14},\n\tcopyright = {All rights reserved},\n\tissn = {2075-4426},\n\tdoi = {10.3390/jpm14030285},\n\tabstract = {Personalized medicine is defined as a medical model using the characterization of individuals' phenotypes and genotypes (e [...].},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {Journal of Personalized Medicine},\n\tauthor = {Ceci, Adriana and Kountouris, Petros and Didio, Antonella and Bonifazi, Fedele},\n\tmonth = mar,\n\tyear = {2024},\n\tpmid = {38541027},\n\tpages = {285},\n}\n\n

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\n \n\n \n \n \n \n \n Unravelling the Complexity of the +33 C\\textgreaterG [HBB:c.-18C\\textgreaterG] Variant in Beta Thalassemia.\n \n \n \n\n\n \n Stephanou, C., Petrou, M., Kountouris, P., Makariou, C., Christou, S., Hadjigavriel, M., Kleanthous, M., & Papasavva, T.\n\n\n \n\n\n\n Biomedicines, 12(2): 296. January 2024.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

\n

@article{stephanou_unravelling_2024,\n\ttitle = {Unravelling the {Complexity} of the +33 {C}{\\textgreater}{G} [{HBB}:c.-{18C}{\\textgreater}{G}] {Variant} in {Beta} {Thalassemia}},\n\tvolume = {12},\n\tcopyright = {All rights reserved},\n\tissn = {2227-9059},\n\tshorttitle = {Unravelling the {Complexity} of the +33 {C}{\\textgreater}{G} [{HBB}},\n\tdoi = {10.3390/biomedicines12020296},\n\tabstract = {The +33 C{\\textgreater}G variant [NM\\_000518.5(HBB):c.-18C{\\textgreater}G] in the 5' untranslated region (UTR) of the β-globin gene is described in the literature as both mild and silent, while it causes a phenotype of thalassemia intermedia in the presence of a severe β-thalassemia allele. Despite its potential clinical significance, the determination of its pathogenicity according to established standards requires a greater number of published cases and co-segregation evidence than what is currently available. The present study provides an extensive phenotypic characterization of +33 C{\\textgreater}G using 26 heterozygous and 11 compound heterozygous novel cases detected in Cyprus and employs computational predictors (CADD, RegulomeDB) to better understand its impact on clinical severity. Genotype identification of globin gene variants, including α- and δ-thalassemia determinants, and rs7482144 (XmnI) was carried out using Sanger sequencing, gap-PCR, and restriction enzyme digestion methods. The heterozygous state of +33 C{\\textgreater}G had a silent phenotype without apparent microcytosis or hypochromia, while compound heterozygosity with a β+ or β0 allele had a spectrum of clinical phenotypes. Awareness of the +33 C{\\textgreater}G is required across Mediterranean populations where β-thalassemia is frequent, particularly in Cyprus, with significant relevance in population screening and fetal diagnostic applications.},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {Biomedicines},\n\tauthor = {Stephanou, Coralea and Petrou, Miranda and Kountouris, Petros and Makariou, Christiana and Christou, Soteroula and Hadjigavriel, Michael and Kleanthous, Marina and Papasavva, Thessalia},\n\tmonth = jan,\n\tyear = {2024},\n\tpmid = {38397898},\n\tpmcid = {PMC10886608},\n\tkeywords = {5′UTR, HBB, genotype/phenotype correlation, silent variant, β-thalassemia intermedia},\n\tpages = {296},\n}\n\n

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\n \n 2023\n \n \n (8)\n \n \n

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\n \n\n \n \n \n \n \n Evaluation of Mono- and Bi-Functional GLOBE-Based Vectors for Therapy of β-Thalassemia by HBBAS3 Gene Addition and Mutation-Specific RNA Interference.\n \n \n \n\n\n \n Koniali, L., Flouri, C., Kostopoulou, M. I., Papaioannou, N. Y., Papasavva, P. L., Naiisseh, B., Stephanou, C., Demetriadou, A., Sitarou, M., Christou, S., Antoniou, M. N., Kleanthous, M., Patsali, P., & Lederer, C. W.\n\n\n \n\n\n\n Cells, 12(24): 2848. December 2023.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{koniali_evaluation_2023,\n\ttitle = {Evaluation of {Mono}- and {Bi}-{Functional} {GLOBE}-{Based} {Vectors} for {Therapy} of β-{Thalassemia} by {HBBAS3} {Gene} {Addition} and {Mutation}-{Specific} {RNA} {Interference}},\n\tvolume = {12},\n\tissn = {2073-4409},\n\tdoi = {10.3390/cells12242848},\n\tabstract = {Therapy via the gene addition of the anti-sickling βAS3-globin transgene is potentially curative for all β-hemoglobinopathies and therefore of particular clinical and commercial interest. This study investigates GLOBE-based lentiviral vectors (LVs) for βAS3-globin addition and evaluates strategies for an increased β-like globin expression without vector dose escalation. First, we report the development of a GLOBE-derived LV, GLV2-βAS3, which, compared to its parental vector, adds anti-sickling action and a transcription-enhancing 848-bp transcription terminator element, retains high vector titers and allows for superior β-like globin expression in primary patient-derived hematopoietic stem and progenitor cells (HSPCs). Second, prompted by our previous correction of HBBIVSI-110(G{\\textgreater}A) thalassemia based on RNApol(III)-driven shRNAs in mono- and combination therapy, we analyzed a series of novel LVs for the RNApol(II)-driven constitutive or late-erythroid expression of HBBIVSI-110(G{\\textgreater}A)-specific miRNA30-embedded shRNAs (shRNAmiR). This included bifunctional LVs, allowing for concurrent βAS3-globin expression. LVs were initially compared for their ability to achieve high β-like globin expression in HBBIVSI-110(G{\\textgreater}A)-transgenic cells, before the evaluation of shortlisted candidate LVs in HBBIVSI-110(G{\\textgreater}A)-homozygous HSPCs. The latter revealed that β-globin promoter-driven designs for monotherapy with HBBIVSI-110(G{\\textgreater}A)-specific shRNAmiRs only marginally increased β-globin levels compared to untransduced cells, whereas bifunctional LVs combining miR30-shRNA with βAS3-globin expression showed disease correction similar to that achieved by the parental GLV2-βAS3 vector. Our results establish the feasibility of high titers for LVs containing the full HBB transcription terminator, emphasize the importance of the HBB terminator for the high-level expression of HBB-like transgenes, qualify the therapeutic utility of late-erythroid HBBIVSI-110(G{\\textgreater}A)-specific miR30-shRNA expression and highlight the exceptional potential of GLV2-βAS3 for the treatment of severe β-hemoglobinopathies.},\n\tlanguage = {eng},\n\tnumber = {24},\n\tjournal = {Cells},\n\tauthor = {Koniali, Lola and Flouri, Christina and Kostopoulou, Markela I. and Papaioannou, Nikoletta Y. and Papasavva, Panayiota L. and Naiisseh, Basma and Stephanou, Coralea and Demetriadou, Anthi and Sitarou, Maria and Christou, Soteroula and Antoniou, Michael N. and Kleanthous, Marina and Patsali, Petros and Lederer, Carsten W.},\n\tmonth = dec,\n\tyear = {2023},\n\tpmid = {38132168},\n\tpmcid = {PMC10741507},\n\tkeywords = {Genetic Therapy, Genetic Vectors, Hemoglobinopathies, Humans, Mutation, RNA Interference, RNA interference, RNA, Small Interfering, beta-Globins, beta-Thalassemia, gene therapy, hemoglobinopathy, lentiviral vector, shRNAmiR, sickle cell anemia, thalassemia},\n\tpages = {2848},\n}\n\n

\n

\n\n\n

\n Therapy via the gene addition of the anti-sickling βAS3-globin transgene is potentially curative for all β-hemoglobinopathies and therefore of particular clinical and commercial interest. This study investigates GLOBE-based lentiviral vectors (LVs) for βAS3-globin addition and evaluates strategies for an increased β-like globin expression without vector dose escalation. First, we report the development of a GLOBE-derived LV, GLV2-βAS3, which, compared to its parental vector, adds anti-sickling action and a transcription-enhancing 848-bp transcription terminator element, retains high vector titers and allows for superior β-like globin expression in primary patient-derived hematopoietic stem and progenitor cells (HSPCs). Second, prompted by our previous correction of HBBIVSI-110(G\\textgreaterA) thalassemia based on RNApol(III)-driven shRNAs in mono- and combination therapy, we analyzed a series of novel LVs for the RNApol(II)-driven constitutive or late-erythroid expression of HBBIVSI-110(G\\textgreaterA)-specific miRNA30-embedded shRNAs (shRNAmiR). This included bifunctional LVs, allowing for concurrent βAS3-globin expression. LVs were initially compared for their ability to achieve high β-like globin expression in HBBIVSI-110(G\\textgreaterA)-transgenic cells, before the evaluation of shortlisted candidate LVs in HBBIVSI-110(G\\textgreaterA)-homozygous HSPCs. The latter revealed that β-globin promoter-driven designs for monotherapy with HBBIVSI-110(G\\textgreaterA)-specific shRNAmiRs only marginally increased β-globin levels compared to untransduced cells, whereas bifunctional LVs combining miR30-shRNA with βAS3-globin expression showed disease correction similar to that achieved by the parental GLV2-βAS3 vector. Our results establish the feasibility of high titers for LVs containing the full HBB transcription terminator, emphasize the importance of the HBB terminator for the high-level expression of HBB-like transgenes, qualify the therapeutic utility of late-erythroid HBBIVSI-110(G\\textgreaterA)-specific miR30-shRNA expression and highlight the exceptional potential of GLV2-βAS3 for the treatment of severe β-hemoglobinopathies.\n

\n\n\n

\n \n\n \n \n \n \n \n Progress and harmonization of gene editing to treat human diseases: Proceeding of COST Action CA21113 GenE-HumDi.\n \n \n \n\n\n \n Cavazza, A., Hendel, A., Bak, R. O., Rio, P., Güell, M., Lainšček, D., Arechavala-Gomeza, V., Peng, L., Hapil, F. Z., Harvey, J., Ortega, F. G., Gonzalez-Martinez, C., Lederer, C. W., Mikkelsen, K., Gasiunas, G., Kalter, N., Gonçalves, M. A. F. V., Petersen, J., Garanto, A., Montoliu, L., Maresca, M., Seemann, S. E., Gorodkin, J., Mazini, L., Sanchez, R., Rodriguez-Madoz, J. R., Maldonado-Pérez, N., Laura, T., Schmueck-Henneresse, M., Maccalli, C., Grünewald, J., Carmona, G., Kachamakova-Trojanowska, N., Miccio, A., Martin, F., Turchiano, G., Cathomen, T., Luo, Y., Tsai, S. Q., Benabdellah, K., & COST Action CA21113\n\n\n \n\n\n\n Molecular Therapy. Nucleic Acids, 34: 102066. December 2023.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{cavazza_progress_2023,\n\ttitle = {Progress and harmonization of gene editing to treat human diseases: {Proceeding} of {COST} {Action} {CA21113} {GenE}-{HumDi}},\n\tvolume = {34},\n\tissn = {2162-2531},\n\tshorttitle = {Progress and harmonization of gene editing to treat human diseases},\n\tdoi = {10.1016/j.omtn.2023.102066},\n\tabstract = {The European Cooperation in Science and Technology (COST) is an intergovernmental organization dedicated to funding and coordinating scientific and technological research in Europe, fostering collaboration among researchers and institutions across countries. Recently, COST Action funded the "Genome Editing to treat Human Diseases" (GenE-HumDi) network, uniting various stakeholders such as pharmaceutical companies, academic institutions, regulatory agencies, biotech firms, and patient advocacy groups. GenE-HumDi's primary objective is to expedite the application of genome editing for therapeutic purposes in treating human diseases. To achieve this goal, GenE-HumDi is organized in several working groups, each focusing on specific aspects. These groups aim to enhance genome editing technologies, assess delivery systems, address safety concerns, promote clinical translation, and develop regulatory guidelines. The network seeks to establish standard procedures and guidelines for these areas to standardize scientific practices and facilitate knowledge sharing. Furthermore, GenE-HumDi aims to communicate its findings to the public in accessible yet rigorous language, emphasizing genome editing's potential to revolutionize the treatment of many human diseases. The inaugural GenE-HumDi meeting, held in Granada, Spain, in March 2023, featured presentations from experts in the field, discussing recent breakthroughs in delivery methods, safety measures, clinical translation, and regulatory aspects related to gene editing.},\n\tlanguage = {eng},\n\tjournal = {Molecular Therapy. Nucleic Acids},\n\tauthor = {Cavazza, Alessia and Hendel, Ayal and Bak, Rasmus O. and Rio, Paula and Güell, Marc and Lainšček, Duško and Arechavala-Gomeza, Virginia and Peng, Ling and Hapil, Fatma Zehra and Harvey, Joshua and Ortega, Francisco G. and Gonzalez-Martinez, Coral and Lederer, Carsten W. and Mikkelsen, Kasper and Gasiunas, Giedrius and Kalter, Nechama and Gonçalves, Manuel A. F. V. and Petersen, Julie and Garanto, Alejandro and Montoliu, Lluis and Maresca, Marcello and Seemann, Stefan E. and Gorodkin, Jan and Mazini, Loubna and Sanchez, Rosario and Rodriguez-Madoz, Juan R. and Maldonado-Pérez, Noelia and Laura, Torella and Schmueck-Henneresse, Michael and Maccalli, Cristina and Grünewald, Julian and Carmona, Gloria and Kachamakova-Trojanowska, Neli and Miccio, Annarita and Martin, Francisco and Turchiano, Giandomenico and Cathomen, Toni and Luo, Yonglun and Tsai, Shengdar Q. and Benabdellah, Karim and {COST Action CA21113}},\n\tmonth = dec,\n\tyear = {2023},\n\tpmid = {38034032},\n\tpmcid = {PMC10685310},\n\tkeywords = {COST, European Cooperation in Science and Technology, GenE-HumDi, MT: RNA/DNA Editing, base editors, delivery systems, genome editing, regulatory guidelines},\n\tpages = {102066},\n}\n\n

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\n \n\n \n \n \n \n \n Submitting Novel Globin Gene Variants to Hemoglobin.\n \n \n \n\n\n \n Harteveld, C. L., Patrinos, G. P., Traeger-Synodinos, J., Kountouris, P., Bento, C., & Adekile, A.\n\n\n \n\n\n\n Hemoglobin, 47(4): 135–136. November 2023.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{harteveld_submitting_2023,\n\ttitle = {Submitting {Novel} {Globin} {Gene} {Variants} to {Hemoglobin}},\n\tvolume = {47},\n\tcopyright = {All rights reserved},\n\tissn = {1532-432X},\n\tdoi = {10.1080/03630269.2023.2258618},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {Hemoglobin},\n\tauthor = {Harteveld, Cornelis L. and Patrinos, George P. and Traeger-Synodinos, Joanne and Kountouris, Petros and Bento, Celeste and Adekile, Adekunle},\n\tmonth = nov,\n\tyear = {2023},\n\tpmid = {37920883},\n\tpages = {135--136},\n}\n\n

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\n \n\n \n \n \n \n \n \n Editorial: Insights in thalassemia: from genomics to clinical practice.\n \n \n \n \n\n\n \n Silao, C. L. T., Latiff, Z. A., Kountouris, P., & Zilfalil, B. A.\n\n\n \n\n\n\n Frontiers in Pediatrics, 11: 1222946. July 2023.\n \n\n\n\n
\n\n\n\n \n \n $\"Editorial:$ Paper\n \n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{silao_editorial_2023,\n\ttitle = {Editorial: {Insights} in thalassemia: from genomics to clinical practice},\n\tvolume = {11},\n\tcopyright = {All rights reserved},\n\tissn = {2296-2360},\n\tshorttitle = {Editorial},\n\turl = {https://www.frontiersin.org/articles/10.3389/fped.2023.1222946/full},\n\tdoi = {10.3389/fped.2023.1222946},\n\turldate = {2023-07-26},\n\tjournal = {Frontiers in Pediatrics},\n\tauthor = {Silao, Catherine Lynn T. and Latiff, Zarina Abdul and Kountouris, Petros and Zilfalil, Bin Alwi},\n\tmonth = jul,\n\tyear = {2023},\n\tpages = {1222946},\n}\n\n

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\n \n\n \n \n \n \n \n Sickle cell disease landscape and challenges in the EU: the ERN-EuroBloodNet perspective.\n \n \n \n\n\n \n Mañú Pereira, M. D. M., Colombatti, R., Alvarez, F., Bartolucci, P., Bento, C., Brunetta, A. L., Cela, E., Christou, S., Collado, A., de Montalembert, M., Dedeken, L., Fenaux, P., Galacteros, F., Glenthøj, A., Gutiérrez Valle, V., Kattamis, A., Kunz, J., Lobitz, S., McMahon, C., Pellegrini, M., Reidel, S., Russo, G., Santos Freire, M., van Beers, E., Kountouris, P., & Gulbis, B.\n\n\n \n\n\n\n The Lancet. Haematology,S2352–3026(23)00182–5. July 2023.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{manu_pereira_sickle_2023,\n\ttitle = {Sickle cell disease landscape and challenges in the {EU}: the {ERN}-{EuroBloodNet} perspective},\n\tcopyright = {All rights reserved},\n\tissn = {2352-3026},\n\tshorttitle = {Sickle cell disease landscape and challenges in the {EU}},\n\tdoi = {10.1016/S2352-3026(23)00182-5},\n\tabstract = {Sickle cell disease is a hereditary multiorgan disease that is considered rare in the EU. In 2017, the Rare Diseases Plan was implemented within the EU and 24 European Reference Networks (ERNs) were created, including the ERN on Rare Haematological Diseases (ERN-EuroBloodNet), dedicated to rare haematological diseases. This EU initiative has made it possible to accentuate existing collaborations and create new ones. The project also made it possible to list all the needs of people with rare haematological diseases not yet covered health-care providers in the EU to allow optimised care of individuals with rare pathologies, including sickle cell disease. This Viewpoint is the result of joint work within 12 EU member states (ie, Belgium, Cyprus, Denmark, France, Germany, Greece, Ireland, Italy, Portugal, Spain, Sweden, and The Netherlands), all members of the ERN-EuroBloodNet. We describe the role of the ERN-EuroBloodNet to improve the overall approach to and the management of individuals with sickle cell disease in the EU through specific on the pooling of expertise, knowledge, and best practices; the development of training and education programmes; the strategy for systematic gathering and standardisation of clinical data; and its reuse in clinical research. Epidemiology and research strategies from ongoing implementation of the Rare Anaemia Disorders European Epidemiological Platform is depicted.},\n\tlanguage = {eng},\n\tjournal = {The Lancet. Haematology},\n\tauthor = {Mañú Pereira, María Del Mar and Colombatti, Raffaella and Alvarez, Federico and Bartolucci, Pablo and Bento, Celeste and Brunetta, Angelo Loris and Cela, Elena and Christou, Soteroula and Collado, Anna and de Montalembert, Mariane and Dedeken, Laurence and Fenaux, Pierre and Galacteros, Frédéric and Glenthøj, Andreas and Gutiérrez Valle, Victoria and Kattamis, Antonis and Kunz, Joachim and Lobitz, Stephan and McMahon, Corrina and Pellegrini, Mariangela and Reidel, Sara and Russo, Giovanna and Santos Freire, Miriam and van Beers, Eduard and Kountouris, Petros and Gulbis, Béatrice},\n\tmonth = jul,\n\tyear = {2023},\n\tpmid = {37451300},\n\tpages = {S2352--3026(23)00182--5},\n}\n\n

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\n \n\n \n \n \n \n \n IthaPhen: An Interactive Database of Genotype-Phenotype Data for Hemoglobinopathies.\n \n \n \n\n\n \n Xenophontos, M., Minaidou, A., Stephanou, C., Tamana, S., Kleanthous, M., & Kountouris, P.\n\n\n \n\n\n\n HemaSphere, 7(7): e922. July 2023.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{xenophontos_ithaphen_2023,\n\ttitle = {{IthaPhen}: {An} {Interactive} {Database} of {Genotype}-{Phenotype} {Data} for {Hemoglobinopathies}},\n\tvolume = {7},\n\tcopyright = {All rights reserved},\n\tissn = {2572-9241},\n\tshorttitle = {{IthaPhen}},\n\tdoi = {10.1097/HS9.0000000000000922},\n\tlanguage = {eng},\n\tnumber = {7},\n\tjournal = {HemaSphere},\n\tauthor = {Xenophontos, Maria and Minaidou, Anna and Stephanou, Coralea and Tamana, Stella and Kleanthous, Marina and Kountouris, Petros},\n\tmonth = jul,\n\tyear = {2023},\n\tpmid = {37359188},\n\tpmcid = {PMC10289560},\n\tpages = {e922},\n}\n\n

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\n \n\n \n \n \n \n \n High-efficiency editing in hematopoietic stem cells and the HUDEP-2 cell line based on in vitro mRNA synthesis.\n \n \n \n\n\n \n Papaioannou, N. Y., Patsali, P., Naiisseh, B., Papasavva, P. L., Koniali, L., Kurita, R., Nakamura, Y., Christou, S., Sitarou, M., Mussolino, C., Cathomen, T., Kleanthous, M., & Lederer, C. W.\n\n\n \n\n\n\n Frontiers in Genome Editing, 5: 1141618. 2023.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{papaioannou_high-efficiency_2023,\n\ttitle = {High-efficiency editing in hematopoietic stem cells and the {HUDEP}-2 cell line based on in vitro {mRNA} synthesis},\n\tvolume = {5},\n\tissn = {2673-3439},\n\tdoi = {10.3389/fgeed.2023.1141618},\n\tabstract = {Introduction: Genome editing tools, such as CRISPR/Cas, TALE nucleases and, more recently, double-strand-break-independent editors, have been successfully used for gene therapy and reverse genetics. Among various challenges in the field, tolerable and efficient delivery of editors to target cells and sites, as well as independence from commercially available tools for flexibility and fast adoption of new editing technology are the most pressing. For many hematopoietic research applications, primary CD34+ cells and the human umbilical cord-derived progenitor erythroid 2 (HUDEP-2) cell line are highly informative substrates and readily accessible for in vitro manipulation. Moreover, ex vivo editing of CD34+ cells has immediate therapeutic relevance. Both cell types are sensitive to standard transfection procedures and reagents, such as lipofection with plasmid DNA, calling for more suitable methodology in order to achieve high efficiency and tolerability of editing with editors of choice. These challenges can be addressed by RNA delivery, either as a mixture of guide RNA and mRNA for CRISRP/Cas-based systems or as a mixture of mRNAs for TALENs. Compared to ribonucleoproteins or proteins, RNA as vector creates flexibility by removing dependence on commercial availability or laborious in-house preparations of novel editor proteins. Compared to DNA, RNA is less toxic and by obviating nuclear transcription and export of mRNA offers faster kinetics and higher editing efficiencies. Methods: Here, we detail an in vitro transcription protocol based on plasmid DNA templates with the addition of Anti-Reverse Cap Analog (ARCA) using T7 RNA polymerase, and poly (A) tailing using poly (A) polymerase, combined with nucleofection of HUDEP-2 and patient-derived CD34+ cells. Our protocol for RNA-based delivery employs widely available reagents and equipment and can easily be adopted for universal in vitro delivery of genome editing tools. Results and Discussion: Drawing on a common use case, we employ the protocol to target a β-globin mutation and to reactivate γ-globin expression as two potential therapies for β-hemoglobinopathies, followed by erythroid differentiation and functional analyses. Our protocol allows high editing efficiencies and unimpaired cell viability and differentiation, with scalability, suitability for functional assessment of editing outcomes and high flexibility in the application to different editors.},\n\tlanguage = {eng},\n\tjournal = {Frontiers in Genome Editing},\n\tauthor = {Papaioannou, Nikoletta Y. and Patsali, Petros and Naiisseh, Basma and Papasavva, Panayiota L. and Koniali, Lola and Kurita, Ryo and Nakamura, Yukio and Christou, Soteroula and Sitarou, Maria and Mussolino, Claudio and Cathomen, Toni and Kleanthous, Marina and Lederer, Carsten W.},\n\tyear = {2023},\n\tpmid = {36969374},\n\tpmcid = {PMC10030607},\n\tkeywords = {CRISPR/Cas, base editor, genome editing, hematopoietic, in vitro transcription, mRNA},\n\tpages = {1141618},\n}\n\n

\n

\n\n\n

\n Introduction: Genome editing tools, such as CRISPR/Cas, TALE nucleases and, more recently, double-strand-break-independent editors, have been successfully used for gene therapy and reverse genetics. Among various challenges in the field, tolerable and efficient delivery of editors to target cells and sites, as well as independence from commercially available tools for flexibility and fast adoption of new editing technology are the most pressing. For many hematopoietic research applications, primary CD34+ cells and the human umbilical cord-derived progenitor erythroid 2 (HUDEP-2) cell line are highly informative substrates and readily accessible for in vitro manipulation. Moreover, ex vivo editing of CD34+ cells has immediate therapeutic relevance. Both cell types are sensitive to standard transfection procedures and reagents, such as lipofection with plasmid DNA, calling for more suitable methodology in order to achieve high efficiency and tolerability of editing with editors of choice. These challenges can be addressed by RNA delivery, either as a mixture of guide RNA and mRNA for CRISRP/Cas-based systems or as a mixture of mRNAs for TALENs. Compared to ribonucleoproteins or proteins, RNA as vector creates flexibility by removing dependence on commercial availability or laborious in-house preparations of novel editor proteins. Compared to DNA, RNA is less toxic and by obviating nuclear transcription and export of mRNA offers faster kinetics and higher editing efficiencies. Methods: Here, we detail an in vitro transcription protocol based on plasmid DNA templates with the addition of Anti-Reverse Cap Analog (ARCA) using T7 RNA polymerase, and poly (A) tailing using poly (A) polymerase, combined with nucleofection of HUDEP-2 and patient-derived CD34+ cells. Our protocol for RNA-based delivery employs widely available reagents and equipment and can easily be adopted for universal in vitro delivery of genome editing tools. Results and Discussion: Drawing on a common use case, we employ the protocol to target a β-globin mutation and to reactivate γ-globin expression as two potential therapies for β-hemoglobinopathies, followed by erythroid differentiation and functional analyses. Our protocol allows high editing efficiencies and unimpaired cell viability and differentiation, with scalability, suitability for functional assessment of editing outcomes and high flexibility in the application to different editors.\n

\n\n\n

\n \n\n \n \n \n \n \n \n Challenges and Opportunities of Precision Medicine in Sickle Cell Disease: Novel European Approach by GenoMed4All Consortium and ERN-EuroBloodNet.\n \n \n \n \n\n\n \n Collado, A., Boaro, M. P., van der Veen, S., Idrizovic, A., Biemond, B. J., Beneitez Pastor, D., Ortuño, A., Cela, E., Ruiz-Llobet, A., Bartolucci, P., de Montalembert, M., Castellani, G., Biondi, R., Manara, R., Sanavia, T., Fariselli, P., Kountouris, P., Kleanthous, M., Alvarez, F., Zazo, S., Colombatti, R., van Beers, E. J., & Mañú-Pereira, M. d. M.\n\n\n \n\n\n\n HemaSphere, 7(3): e844. March 2023.\n \n\n\n\n
\n\n\n\n \n \n $\"Challenges$ Paper\n \n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{collado_challenges_2023,\n\ttitle = {Challenges and {Opportunities} of {Precision} {Medicine} in {Sickle} {Cell} {Disease}: {Novel} {European} {Approach} by {GenoMed4All} {Consortium} and {ERN}-{EuroBloodNet}},\n\tvolume = {7},\n\tcopyright = {All rights reserved},\n\tissn = {2572-9241},\n\tshorttitle = {Challenges and {Opportunities} of {Precision} {Medicine} in {Sickle} {Cell} {Disease}},\n\turl = {https://journals.lww.com/10.1097/HS9.0000000000000844},\n\tdoi = {10.1097/HS9.0000000000000844},\n\tlanguage = {en},\n\tnumber = {3},\n\turldate = {2023-02-23},\n\tjournal = {HemaSphere},\n\tauthor = {Collado, Anna and Boaro, Maria Paola and van der Veen, Sigrid and Idrizovic, Amira and Biemond, Bart J. and Beneitez Pastor, David and Ortuño, Ana and Cela, Elena and Ruiz-Llobet, Anna and Bartolucci, Pablo and de Montalembert, Marianne and Castellani, Gastone and Biondi, Riccardo and Manara, Renzo and Sanavia, Tiziana and Fariselli, Piero and Kountouris, Petros and Kleanthous, Marina and Alvarez, Federico and Zazo, Santiago and Colombatti, Raffaella and van Beers, Eduard J. and Mañú-Pereira, María del Mar},\n\tmonth = mar,\n\tyear = {2023},\n\tpages = {e844},\n}\n\n

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\n \n 2022\n \n \n (5)\n \n \n

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\n \n\n \n \n \n \n \n Adapting the ACMG/AMP variant classification framework: A perspective from the ClinGen Hemoglobinopathy Variant Curation Expert Panel.\n \n \n \n\n\n \n Kountouris, P., Stephanou, C., Lederer, C. W., Traeger-Synodinos, J., Bento, C., Harteveld, C. L., Fylaktou, E., Koopmann, T. T., Halim-Fikri, H., Michailidou, K., Nfonsam, L. E., Waye, J. S., Zilfalil, B. A., Kleanthous, M., & ClinGen Hemoglobinopathy Variant Curation Expert Panel\n\n\n \n\n\n\n Human Mutation, 43(8): 1089–1096. August 2022.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kountouris_adapting_2022,\n\ttitle = {Adapting the {ACMG}/{AMP} variant classification framework: {A} perspective from the {ClinGen} {Hemoglobinopathy} {Variant} {Curation} {Expert} {Panel}},\n\tvolume = {43},\n\tissn = {1098-1004},\n\tshorttitle = {Adapting the {ACMG}/{AMP} variant classification framework},\n\tdoi = {10.1002/humu.24280},\n\tabstract = {Accurate and consistent interpretation of sequence variants is integral to the delivery of safe and reliable diagnostic genetic services. To standardize the interpretation process, in 2015, the American College of Medical Genetics and Genomics (ACMG) and the Association for Molecular Pathology (AMP) published a joint guideline based on a set of shared standards for the classification of variants in Mendelian diseases. The generality of these standards and their subjective interpretation between laboratories has prompted efforts to reduce discordance of variant classifications, with a focus on the expert specification of the ACMG/AMP guidelines for individual genes or diseases. Herein, we describe our experience as a ClinGen Variant Curation Expert Panel to adapt the ACMG/AMP criteria for the classification of variants in three globin genes (HBB, HBA2, and HBA1) related to recessively inherited hemoglobinopathies, including five evidence categories, as use cases demonstrating the process of specification and the underlying rationale.},\n\tlanguage = {eng},\n\tnumber = {8},\n\tjournal = {Human Mutation},\n\tauthor = {Kountouris, Petros and Stephanou, Coralea and Lederer, Carsten W. and Traeger-Synodinos, Joanne and Bento, Celeste and Harteveld, Cornelis L. and Fylaktou, Eirini and Koopmann, Tamara T. and Halim-Fikri, Hashim and Michailidou, Kyriaki and Nfonsam, Landry E. and Waye, John S. and Zilfalil, Bin A. and Kleanthous, Marina and {ClinGen Hemoglobinopathy Variant Curation Expert Panel}},\n\tmonth = aug,\n\tyear = {2022},\n\tpmid = {34510646},\n\tpmcid = {PMC9545675},\n\tkeywords = {ACMG/AMP criteria, ClinGen VCEP, Genetic Testing, Genetic Variation, Genome, Human, Hemoglobinopathies, Humans, Pathology, Molecular, United States, globin gene variants, hemoglobinopathy, variant classification},\n\tpages = {1089--1096},\n}\n\n

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\n \n\n \n \n \n \n \n Global Globin Network Consensus Paper: Classification and Stratified Roadmaps for Improved Thalassaemia Care and Prevention in 32 Countries.\n \n \n \n\n\n \n Halim-Fikri, B. H., Lederer, C. W., Baig, A. A., Mat-Ghani, S. N. A., Syed-Hassan, S. R., Yusof, W., Abdul Rashid, D., Azman, N. F., Fucharoen, S., Panigoro, R., Silao, C. L. T., Viprakasit, V., Jalil, N., Mohd Yasin, N., Bahar, R., Selvaratnam, V., Mohamad, N., Nik Hassan, N. N., Esa, E., Krause, A., Robinson, H., Hasler, J., Stephanou, C., Raja-Sabudin, R., Elion, J., El-Kamah, G., Coviello, D., Yusoff, N., Abdul Latiff, Z., Arnold, C., Burn, J., Kountouris, P., Kleanthous, M., Ramesar, R., Zilfalil, B. A., & on behalf of the Global Globin Network (GGN)\n\n\n \n\n\n\n Journal of Personalized Medicine, 12(4): 552. March 2022.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{halim-fikri_global_2022,\n\ttitle = {Global {Globin} {Network} {Consensus} {Paper}: {Classification} and {Stratified} {Roadmaps} for {Improved} {Thalassaemia} {Care} and {Prevention} in 32 {Countries}},\n\tvolume = {12},\n\tissn = {2075-4426},\n\tshorttitle = {Global {Globin} {Network} {Consensus} {Paper}},\n\tdoi = {10.3390/jpm12040552},\n\tabstract = {The Global Globin Network (GGN) is a project-wide initiative of the Human Variome/Global Variome Project (HVP) focusing on haemoglobinopathies to build the capacity for genomic diagnosis, clinical services, and research in low- and middle-income countries. At present, there is no framework to evaluate the improvement of care, treatment, and prevention of thalassaemia and other haemoglobinopathies globally, despite thalassaemia being one of the most common monogenic diseases worldwide. Here, we propose a universally applicable system for evaluating and grouping countries based on qualitative indicators according to the quality of care, treatment, and prevention of haemoglobinopathies. We also apply this system to GGN countries as proof of principle. To this end, qualitative indicators were extracted from the IthaMaps database of the ITHANET portal, which allowed four groups of countries (A, B, C, and D) to be defined based on major qualitative indicators, supported by minor qualitative indicators for countries with limited resource settings and by the overall haemoglobinopathy carrier frequency for the target countries of immigration. The proposed rubrics and accumulative scores will help analyse the performance and improvement of care, treatment, and prevention of haemoglobinopathies in the GGN and beyond. Our proposed criteria complement future data collection from GGN countries to help monitor the quality of services for haemoglobinopathies, provide ongoing estimates for services and epidemiology in GGN countries, and note the contribution of the GGN to a local and global reduction of disease burden.},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {Journal of Personalized Medicine},\n\tauthor = {Halim-Fikri, Bin Hashim and Lederer, Carsten W. and Baig, Atif Amin and Mat-Ghani, Siti Nor Assyuhada and Syed-Hassan, Sharifah-Nany Rahayu-Karmilla and Yusof, Wardah and Abdul Rashid, Diana and Azman, Nurul Fatihah and Fucharoen, Suthat and Panigoro, Ramdan and Silao, Catherine Lynn T. and Viprakasit, Vip and Jalil, Norunaluwar and Mohd Yasin, Norafiza and Bahar, Rosnah and Selvaratnam, Veena and Mohamad, Norsarwany and Nik Hassan, Nik Norliza and Esa, Ezalia and Krause, Amanda and Robinson, Helen and Hasler, Julia and Stephanou, Coralea and Raja-Sabudin, Raja-Zahratul-Azma and Elion, Jacques and El-Kamah, Ghada and Coviello, Domenico and Yusoff, Narazah and Abdul Latiff, Zarina and Arnold, Chris and Burn, John and Kountouris, Petros and Kleanthous, Marina and Ramesar, Raj and Zilfalil, Bin Alwi and {on behalf of the Global Globin Network (GGN)}},\n\tmonth = mar,\n\tyear = {2022},\n\tpmid = {35455667},\n\tpmcid = {PMC9032232},\n\tkeywords = {Global Globin Network, Hemoglobinopathy VCEP, Human Variome Project, disease burden, epidemiology, haemoglobinopathy, low- and middle-income countries, prevention program, thalassaemia},\n\tpages = {552},\n}\n\n

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\n \n\n \n \n \n \n \n \n Direct Chromosomal Phasing: An Easy and Fast Approach for Broadening Prenatal Diagnostic Applicability.\n \n \n \n \n\n\n \n Byrou, S., Christopoulos, G., Christofides, A., Makariou, C., Ioannou, C., Kleanthous, M., & Papasavva, T.\n\n\n \n\n\n\n Thalassemia Reports, 12(3): 55–72. July 2022.\n \n\n\n\n
\n\n\n\n \n \n $\"Direct$ Paper\n \n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{byrou_direct_2022,\n\ttitle = {Direct {Chromosomal} {Phasing}: {An} {Easy} and {Fast} {Approach} for {Broadening} {Prenatal} {Diagnostic} {Applicability}},\n\tvolume = {12},\n\tissn = {2039-4365},\n\tshorttitle = {Direct {Chromosomal} {Phasing}},\n\turl = {https://www.mdpi.com/2039-4365/12/3/11},\n\tdoi = {10.3390/thalassrep12030011},\n\tabstract = {The assignment of alleles to haplotypes in prenatal diagnostic assays has traditionally depended on family study analyses. However, this prevents the wide application of prenatal diagnosis based on haplotype analysis, especially in countries with dispersed populations. Here, we present an easy and fast approach using Droplet Digital PCR for the direct determination of haplotype blocks, overcoming the necessity for acquiring other family members’ genetic samples. We demonstrate this approach on nine families that were referred to our center for a prenatal diagnosis of β-thalassaemia using four highly polymorphic single nucleotide variations and the most common pathogenic β-thalassaemia variation in our population. Our approach resulted in the successful direct chromosomal phasing and haplotyping for all nine of the families analyzed, demonstrating a complete agreement with the haplotypes that are ascertained based on family trios. The clinical utility of this approach is envisaged to open the application of prenatal diagnosis for β-thalassaemia to all cases, while simultaneously providing a model for extending the prenatal diagnostic application of other monogenic diseases as well.},\n\tlanguage = {en},\n\tnumber = {3},\n\turldate = {2023-02-08},\n\tjournal = {Thalassemia Reports},\n\tauthor = {Byrou, Stefania and Christopoulos, George and Christofides, Agathoklis and Makariou, Christiana and Ioannou, Christiana and Kleanthous, Marina and Papasavva, Thessalia},\n\tmonth = jul,\n\tyear = {2022},\n\tpages = {55--72},\n}\n\n

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\n \n\n \n \n \n \n \n Evaluation of in silico predictors on short nucleotide variants in HBA1, HBA2, and HBB associated with haemoglobinopathies.\n \n \n \n\n\n \n Tamana, S., Xenophontos, M., Minaidou, A., Stephanou, C., Harteveld, C. L., Bento, C., Traeger-Synodinos, J., Fylaktou, I., Yasin, N. M., Abdul Hamid, F. S., Esa, E., Halim-Fikri, H., Zilfalil, B. A., Kakouri, A. C., Kleanthous, M., & Kountouris, P.\n\n\n \n\n\n\n eLife, 11. December 2022.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{tamana_evaluation_2022,\n\ttitle = {Evaluation of in silico predictors on short nucleotide variants in {HBA1}, {HBA2}, and {HBB} associated with haemoglobinopathies.},\n\tvolume = {11},\n\tcopyright = {© 2022, Tamana, Xenophontos et al.},\n\tissn = {2050-084X},\n\tdoi = {10.7554/eLife.79713},\n\tabstract = {Haemoglobinopathies are the commonest monogenic diseases worldwide and are caused by variants in the globin gene clusters. With over 2400 variants detected to  date, their interpretation using the American College of Medical Genetics and  Genomics (ACMG)/Association for Molecular Pathology (AMP) guidelines is  challenging and computational evidence can provide valuable input about their  functional annotation. While many in silico predictors have already been  developed, their performance varies for different genes and diseases. In this  study, we evaluate 31 in silico predictors using a dataset of 1627 variants in  HBA1, HBA2, and HBB. By varying the decision threshold for each tool, we analyse  their performance (a) as binary classifiers of pathogenicity and (b) by using  different non-overlapping pathogenic and benign thresholds for their optimal use  in the ACMG/AMP framework. Our results show that CADD, Eigen-PC, and REVEL are  the overall top performers, with the former reaching moderate strength level for  pathogenic prediction. Eigen-PC and REVEL achieve the highest accuracies for  missense variants, while CADD is also a reliable predictor of non-missense  variants. Moreover, SpliceAI is the top performing splicing predictor, reaching  strong level of evidence, while GERP++ and phyloP are the most accurate  conservation tools. This study provides evidence about the optimal use of  computational tools in globin gene clusters under the ACMG/AMP framework.},\n\tlanguage = {eng},\n\tjournal = {eLife},\n\tauthor = {Tamana, Stella and Xenophontos, Maria and Minaidou, Anna and Stephanou, Coralea and Harteveld, Cornelis L. and Bento, Celeste and Traeger-Synodinos, Joanne and Fylaktou, Irene and Yasin, Norafiza Mohd and Abdul Hamid, Faidatul Syazlin and Esa, Ezalia and Halim-Fikri, Hashim and Zilfalil, Bin Alwi and Kakouri, Andrea C. and Kleanthous, Marina and Kountouris, Petros},\n\tmonth = dec,\n\tyear = {2022},\n\tpmid = {36453528},\n\tpmcid = {PMC9731569},\n\tnote = {Place: England},\n\tkeywords = {*Genomics, *Nucleotides, Humans, Pathology, Molecular, Universities, computational biology, genetics, genomics, globin genes, haemoglobinopathies, human, in silico prediction, systems biology, thalassaemia, variant classification},\n}\n\n

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\n \n\n \n \n \n \n \n A Novel Tool for the Analysis and Detection of Copy Number Variants Associated with Haemoglobinopathies.\n \n \n \n\n\n \n Minaidou, A., Tamana, S., Stephanou, C., Xenophontos, M., Harteveld, C. L., Bento, C., Kleanthous, M., & Kountouris, P.\n\n\n \n\n\n\n International journal of molecular sciences, 23(24). December 2022.\n Place: Switzerland\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{minaidou_novel_2022,\n\ttitle = {A {Novel} {Tool} for the {Analysis} and {Detection} of {Copy} {Number} {Variants} {Associated} with {Haemoglobinopathies}.},\n\tvolume = {23},\n\tcopyright = {All rights reserved},\n\tissn = {1422-0067},\n\tdoi = {10.3390/ijms232415920},\n\tabstract = {Several types of haemoglobinopathies are caused by copy number variants (CNVs). While diagnosis is often based on haematological and biochemical parameters, a  definitive diagnosis requires molecular DNA analysis. In some cases, the  molecular characterisation of large deletions/duplications is challenging and  inconclusive and often requires the use of specific diagnostic procedures, such  as multiplex ligation-dependent probe amplification (MLPA). Herein, we collected  and comprehensively analysed all known CNVs associated with haemoglobinopathies.  The dataset of 291 CNVs was retrieved from the IthaGenes database and was further  manually annotated to specify genomic locations, breakpoints and MLPA probes  relevant for each CNV. We developed IthaCNVs, a publicly available and  easy-to-use online tool that can facilitate the diagnosis of rare and  diagnostically challenging haemoglobinopathy cases attributed to CNVs.  Importantly, it facilitates the filtering of available entries based on the type  of breakpoint information, on specific chromosomal and locus positions, on MLPA  probes, and on affected gene(s). IthaCNVs brings together manually curated  information about CNV genomic locations, functional effects, and information that  can facilitate CNV characterisation through MLPA. It can help laboratory staff  and clinicians confirm suspected diagnosis of CNVs based on molecular DNA  screening and analysis.},\n\tlanguage = {eng},\n\tnumber = {24},\n\tjournal = {International journal of molecular sciences},\n\tauthor = {Minaidou, Anna and Tamana, Stella and Stephanou, Coralea and Xenophontos, Maria and Harteveld, Cornelis L. and Bento, Celeste and Kleanthous, Marina and Kountouris, Petros},\n\tmonth = dec,\n\tyear = {2022},\n\tpmid = {36555557},\n\tpmcid = {PMC9782104},\n\tnote = {Place: Switzerland},\n\tkeywords = {*DNA Copy Number Variations/genetics, *Genome, DNA, Genomics, Humans, MLPA, Multiplex Polymerase Chain Reaction/methods, copy number variants (CNVs), haemoglobinopathies},\n}\n\n

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\n \n 2021\n \n \n (4)\n \n \n

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\n \n\n \n \n \n \n \n Editorial: Mutation-Specific Gene Editing for Blood Disorders.\n \n \n \n\n\n \n Lederer, C. W., Genovese, P., Miccio, A., & Philipsen, S.\n\n\n \n\n\n\n Frontiers in Genome Editing, 3: 761771. 2021.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{lederer_editorial_2021,\n\ttitle = {Editorial: {Mutation}-{Specific} {Gene} {Editing} for {Blood} {Disorders}},\n\tvolume = {3},\n\tissn = {2673-3439},\n\tshorttitle = {Editorial},\n\tdoi = {10.3389/fgeed.2021.761771},\n\tlanguage = {eng},\n\tjournal = {Frontiers in Genome Editing},\n\tauthor = {Lederer, Carsten Werner and Genovese, Pietro and Miccio, Annarita and Philipsen, Sjaak},\n\tyear = {2021},\n\tpmid = {34713270},\n\tpmcid = {PMC8525395},\n\tkeywords = {Base Editing, CRISPR/Cas, Gene Editing, Hematopathology, Hoogsteen, Nanoblades, Perspective, Prime Editing},\n\tpages = {761771},\n}\n\n

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\n \n\n \n \n \n \n \n Regulation of ER Composition and Extent, and Putative Action in Protein Networks by ER/NE Protein TMEM147.\n \n \n \n\n\n \n Maimaris, G., Christodoulou, A., Santama, N., & Lederer, C. W.\n\n\n \n\n\n\n International Journal of Molecular Sciences, 22(19): 10231. September 2021.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{maimaris_regulation_2021,\n\ttitle = {Regulation of {ER} {Composition} and {Extent}, and {Putative} {Action} in {Protein} {Networks} by {ER}/{NE} {Protein} {TMEM147}},\n\tvolume = {22},\n\tissn = {1422-0067},\n\tdoi = {10.3390/ijms221910231},\n\tabstract = {Nuclear envelope (NE) and endoplasmic reticulum (ER) collaborate to control a multitude of nuclear and cytoplasmic actions. In this context, the transmembrane protein TMEM147 localizes to both NE and ER, and through direct and indirect interactions regulates processes as varied as production and transport of multipass membrane proteins, neuronal signaling, nuclear-shape, lamina and chromatin dynamics and cholesterol synthesis. Aiming to delineate the emerging multifunctionality of TMEM147 more comprehensively, we set as objectives, first, to assess potentially more fundamental effects of TMEM147 on the ER and, second, to identify significantly TMEM147-associated cell-wide protein networks and pathways. Quantifying curved and flat ER markers RTN4 and CLIMP63/CKAP4, respectively, we found that TMEM147 silencing causes area and intensity increases for both RTN4 and CLIMP63, and the ER in general, with a profound shift toward flat areas, concurrent with reduction in DNA condensation. Protein network and pathway analyses based on comprehensive compilation of TMEM147 interactors, targets and co-factors then served to manifest novel and established roles for TMEM147. Thus, algorithmically simplified significant pathways reflect TMEM147 function in ribosome binding, oxidoreductase activity, G protein-coupled receptor activity and transmembrane transport, while analysis of protein factors and networks identifies hub proteins and corresponding pathways as potential targets of TMEM147 action and of future functional studies.},\n\tlanguage = {eng},\n\tnumber = {19},\n\tjournal = {International Journal of Molecular Sciences},\n\tauthor = {Maimaris, Giannis and Christodoulou, Andri and Santama, Niovi and Lederer, Carsten Werner},\n\tmonth = sep,\n\tyear = {2021},\n\tpmid = {34638576},\n\tpmcid = {PMC8508377},\n\tkeywords = {Endoplasmic Reticulum, Gene Silencing, HeLa Cells, Humans, Lamin B Receptor, Membrane Proteins, Nogo Proteins, Nuclear Envelope, Protein Interaction Maps, RNA interference, Receptors, Cytoplasmic and Nuclear, Signal Transduction, endoplasmic reticulum, gene ontology, network analysis, nuclear envelope},\n\tpages = {10231},\n}\n\n

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\n \n\n \n \n \n \n \n Therapy Development by Genome Editing of Hematopoietic Stem Cells.\n \n \n \n\n\n \n Lola Koniali, Koniali, L., Lederer, C. W., & Kleanthous, M.\n\n\n \n\n\n\n Cells, 10(6): 1492. June 2021.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{lola_koniali_therapy_2021,\n\ttitle = {Therapy {Development} by {Genome} {Editing} of {Hematopoietic} {Stem} {Cells}},\n\tvolume = {10},\n\tdoi = {10.3390/cells10061492},\n\tabstract = {Accessibility of hematopoietic stem cells (HSCs) for the manipulation and repopulation of the blood and immune systems has placed them at the forefront of cell and gene therapy development. Recent advances in genome-editing tools, in particular for clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) and CRISPR/Cas-derived editing systems, have transformed the gene therapy landscape. Their versatility and the ability to edit genomic sequences and facilitate gene disruption, correction or insertion, have broadened the spectrum of potential gene therapy targets and accelerated the development of potential curative therapies for many rare diseases treatable by transplantation or modification of HSCs. Ongoing developments seek to address efficiency and precision of HSC modification, tolerability of treatment and the distribution and affordability of corresponding therapies. Here, we give an overview of recent progress in the field of HSC genome editing as treatment for inherited disorders and summarize the most significant findings from corresponding preclinical and clinical studies. With emphasis on HSC-based therapies, we also discuss technical hurdles that need to be overcome en route to clinical translation of genome editing and indicate advances that may facilitate routine application beyond the most common disorders.},\n\tnumber = {6},\n\tjournal = {Cells},\n\tauthor = {{Lola Koniali} and Koniali, Lola and Lederer, Carsten W. and Kleanthous, Marina},\n\tmonth = jun,\n\tyear = {2021},\n\tdoi = {10.3390/cells10061492},\n\tpages = {1492},\n}\n\n

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\n \n\n \n \n \n \n \n The International Hemoglobinopathy Research Network (INHERENT): An international initiative to study the role of genetic modifiers in hemoglobinopathies.\n \n \n \n\n\n \n Kountouris, P., Stephanou, C., Archer, N., Bonifazi, F., Giannuzzi, V., Kuo, K. H. M., Maggio, A., Makani, J., Mañú-Pereira, M. D. M., Michailidou, K., Nkya, S., Nnodu, O. E., Trompeter, S., Tshilolo, L., Wonkam, A., Zilfalil, B. A., Inusa, B. P. D., Kleanthous, M., & on behalf of the International Hemoglobinopathy Research Network (INHERENT)\n\n\n \n\n\n\n American Journal of Hematology, 96(11): E416–E420. November 2021.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kountouris_international_2021,\n\ttitle = {The {International} {Hemoglobinopathy} {Research} {Network} ({INHERENT}): {An} international initiative to study the role of genetic modifiers in hemoglobinopathies},\n\tvolume = {96},\n\tissn = {1096-8652},\n\tshorttitle = {The {International} {Hemoglobinopathy} {Research} {Network} ({INHERENT})},\n\tdoi = {10.1002/ajh.26323},\n\tlanguage = {eng},\n\tnumber = {11},\n\tjournal = {American Journal of Hematology},\n\tauthor = {Kountouris, Petros and Stephanou, Coralea and Archer, Natasha and Bonifazi, Fedele and Giannuzzi, Viviana and Kuo, Kevin H. M. and Maggio, Aurelio and Makani, Julie and Mañú-Pereira, María Del Mar and Michailidou, Kyriaki and Nkya, Siana and Nnodu, Obiageli E. and Trompeter, Sara and Tshilolo, Léon and Wonkam, Ambroise and Zilfalil, Bin Alwi and Inusa, Baba P. D. and Kleanthous, Marina and {on behalf of the International Hemoglobinopathy Research Network (INHERENT)}},\n\tmonth = nov,\n\tyear = {2021},\n\tpmid = {34406671},\n\tkeywords = {Anemia, Sickle Cell, Databases, Factual, Genetic Predisposition to Disease, Genetic Variation, Hemoglobinopathies, Humans},\n\tpages = {E416--E420},\n}\n\n

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\n \n 2020\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n TMEM147 interacts with lamin B receptor, regulates its localization and levels, and affects cholesterol homeostasis.\n \n \n \n\n\n \n Christodoulou, A., Maimaris, G., Makrigiorgi, A., Charidemou, E., Lüchtenborg, C., Ververis, A., Georgiou, R., Lederer, C. W., Haffner, C., Brügger, B., & Santama, N.\n\n\n \n\n\n\n Journal of Cell Science, 133(16): jcs245357. August 2020.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{christodoulou_tmem147_2020,\n\ttitle = {{TMEM147} interacts with lamin {B} receptor, regulates its localization and levels, and affects cholesterol homeostasis},\n\tvolume = {133},\n\tissn = {1477-9137},\n\tdoi = {10.1242/jcs.245357},\n\tabstract = {The structurally and functionally complex endoplasmic reticulum (ER) hosts critical processes including lipid synthesis. Here, we focus on the functional characterization of transmembrane protein TMEM147, and report that it localizes at the ER and nuclear envelope in HeLa cells. Silencing of TMEM147 drastically reduces the level of lamin B receptor (LBR) at the inner nuclear membrane and results in mistargeting of LBR to the ER. LBR possesses a modular structure and corresponding bifunctionality, acting in heterochromatin organization via its N-terminus and in cholesterol biosynthesis via its sterol-reductase C-terminal domain. We show that TMEM147 physically interacts with LBR, and that the C-terminus of LBR is essential for their functional interaction. We find that TMEM147 also physically interacts with the key sterol reductase DHCR7, which is involved in cholesterol biosynthesis. Similar to what was seen for LBR, TMEM147 downregulation results in a sharp decline of DHCR protein levels and co-ordinate transcriptional decreases of LBR and DHCR7 expression. Consistent with this, lipidomic analysis upon TMEM147 silencing identified changes in cellular cholesterol levels, cholesteryl ester levels and profile, and in cellular cholesterol uptake, raising the possibility that TMEM147 is an important new regulator of cholesterol homeostasis in cells.This article has an associated First Person interview with the first author of the paper.},\n\tlanguage = {eng},\n\tnumber = {16},\n\tjournal = {Journal of Cell Science},\n\tauthor = {Christodoulou, Andri and Maimaris, Giannis and Makrigiorgi, Andri and Charidemou, Evelina and Lüchtenborg, Christian and Ververis, Antonis and Georgiou, Renos and Lederer, Carsten W. and Haffner, Christof and Brügger, Britta and Santama, Niovi},\n\tmonth = aug,\n\tyear = {2020},\n\tpmid = {32694168},\n\tkeywords = {Cholesterol, ER transmembrane proteins, HeLa Cells, Homeostasis, Humans, LBR, Lamin B Receptor, Membrane Proteins, Nerve Tissue Proteins, Nuclear Envelope, Receptors, Cytoplasmic and Nuclear, Sterol reductases},\n\tpages = {jcs245357},\n}\n\n

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\n \n\n \n \n \n \n \n Relative and Absolute Quantification of Aberrant and Normal Splice Variants in HBBIVSI-110 (G > A) β-Thalassemia.\n \n \n \n\n\n \n Patsali, P., Papasavva, P., Christou, S., Sitarou, M., Antoniou, M., Lederer, C. W., & Kleanthous, M.\n\n\n \n\n\n\n International Journal of Molecular Sciences, 21(18): 6671. 2020.\n MAG ID: 3086658224\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{patsali_relative_2020,\n\ttitle = {Relative and {Absolute} {Quantification} of {Aberrant} and {Normal} {Splice} {Variants} in {HBBIVSI}-110 ({G} \\&\\#62; {A}) β-{Thalassemia}.},\n\tvolume = {21},\n\tdoi = {10.3390/ijms21186671},\n\tabstract = {The β-thalassemias are an increasing challenge to health systems worldwide, caused by absent or reduced β-globin (HBB) production. Of particular frequency in many Western countries is HBBIVSI−110(G {\\textgreater} A) β-thalassemia (HGVS name: HBB:c.93-21G {\\textgreater} A). Its underlying mutation creates an abnormal splice acceptor site in the HBB gene, and while partially retaining normal splicing of HBB, it severely reduces HBB protein expression from the mutant locus and HBB loci in trans. For the assessment of the underlying mechanisms and of therapies targeting β-thalassemia, accurate quantification of aberrant and normal HBB mRNA is essential, but to date, has only been performed by approximate methods. To address this shortcoming, we have developed an accurate, duplex reverse-transcription quantitative PCR assay for the assessment of the ratio and absolute quantities of normal and aberrant mRNA species as a tool for basic and translational research of HBBIVSI−110(G {\\textgreater} A) β-thalassemia. The method was employed here to determine mRNA ratios and quantities in blood and primary cell culture samples and correlate them with HBB protein levels. Moreover, with its immediate utility for β-thalassemia and the mutation in hand, the approach can readily be adopted for analysis of alternative splicing or for quantitative assays of any disease-causing mutation that interferes with normal splicing.},\n\tnumber = {18},\n\tjournal = {International Journal of Molecular Sciences},\n\tauthor = {Patsali, Petros and Papasavva, Panayiota and Christou, Soteroulla and Sitarou, Maria and Antoniou, Michael and Lederer, Carsten W. and Kleanthous, Marina},\n\tyear = {2020},\n\tdoi = {10.3390/ijms21186671},\n\tpmid = {32933098},\n\tnote = {MAG ID: 3086658224},\n\tpages = {6671},\n}\n\n

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\n \n 2019\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n (57)Fe enrichment in mice for β-thalassaemia studies via Mössbauer spectroscopy of blood samples.\n \n \n \n\n\n \n Charitou, G., Tsertos, C., Parpottas, Y., Kleanthous, M., Lederer, C. W., & Phylactides, M.\n\n\n \n\n\n\n European biophysics journal : EBJ, 48(7): 635–643. October 2019.\n Place: Germany\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{charitou_57fe_2019,\n\ttitle = {(57){Fe} enrichment in mice for β-thalassaemia studies via {Mössbauer} spectroscopy of blood samples.},\n\tvolume = {48},\n\tissn = {1432-1017 0175-7571},\n\tdoi = {10.1007/s00249-019-01389-w},\n\tabstract = {In this work, wild-type and heterozygous β-thalassaemic mice were enriched with (57)Fe via gastrointestinal absorption to characterize in greater detail the iron  complexes then identifiable via Mössbauer spectroscopy. The (57)Fe enrichment  method was validated and Mössbauer spectra were obtained at 80 K from blood  samples from wild-type and β-thalassaemic mice at 1, 3, 6, and 9 months of age.  As expected, the haemoglobin levels of the thalassaemic mice were lower than from  normal mice, indicating anaemia. Furthermore, significant amounts of  ferritin-like iron were observed in the thalassaemic mice samples, which  decreased with mouse age, reflecting the pattern of reticulocyte count reduction  reported in the literature.},\n\tlanguage = {eng},\n\tnumber = {7},\n\tjournal = {European biophysics journal : EBJ},\n\tauthor = {Charitou, George and Tsertos, Charalambos and Parpottas, Yannis and Kleanthous, Marina and Lederer, Carsten W. and Phylactides, Marios},\n\tmonth = oct,\n\tyear = {2019},\n\tpmid = {31302726},\n\tnote = {Place: Germany},\n\tkeywords = {*Spectroscopy, Mossbauer, 57Fe enrichment, Animals, Blood, Ferritin, Intestinal Absorption, Iron Isotopes/*metabolism/*pharmacology, Mice, Mice, Inbred C57BL, Mössbauer spectroscopy, beta-Thalassemia/*blood/*metabolism, β-Thalassaemia},\n\tpages = {635--643},\n}\n\n

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\n \n\n \n \n \n \n \n 2'-O-methoxyethyl splice-switching oligos correct splicing from IVS2-745 β-thalassemia patient cells restoring HbA production and chain rebalance.\n \n \n \n\n\n \n Dong, A., Ghiaccio, V., Motta, I., Guo, S., Peralta, R., Freier, S. M., Watt, A., Damle, S., Ikawa, Y., Jarocha, D., Chappell, M., Stephanou, C., Delbini, P., Chen, C., Christou, S., Kleanthous, M., Smith-Whitley, K., Manwani, D., Casu, C., Abdulmalik, O., Cappellini, M. D., Rivella, S., & Breda, L.\n\n\n \n\n\n\n Haematologica, 106(5): 1433–1442. May 2019.\n Place: Italy\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{dong_2-o-methoxyethyl_2019,\n\ttitle = {2'-{O}-methoxyethyl splice-switching oligos correct splicing from {IVS2}-745 β-thalassemia patient cells restoring {HbA} production and chain rebalance.},\n\tvolume = {106},\n\tissn = {1592-8721 0390-6078},\n\tdoi = {10.3324/haematol.2019.226852},\n\tabstract = {β-thalassemia is a disorder caused by altered hemoglobin protein synthesis and affects individuals worldwide. Severe forms of the disease, left untreated, can  result in death before the age of 3 years (1). The standard of care consists of  chronic and costly palliative treatment by blood transfusion combined with iron  chelation. This dual approach suppresses anemia and reduces iron-related  toxicities in patients. Allogeneic bone marrow transplant is an option, but  limited by the availability of a highly compatible HSC donor. While gene therapy  is been explored in several trials, its use is highly limited to developed  regions with centers of excellence and well-established healthcare systems (2).  Hence, there remains a tremendous unmet medical need to develop alternative  treatment strategies for β-thalassemia (3). Occurrence of aberrant splicing is  one of the processes that affects β-globin synthesis in β-thalassemia. The (C{\\textgreater}G)  IVS-2-745 is a splicing mutation within intron 2 of the β-globin gene. It leads  to an aberrantly spliced mRNA that incorporates an intron fragment. This results  in an in-frame premature termination codon that inhibits β-globin production.  Here, we propose the use of uniform 2'-O-methoxyethyl (2'-MOE) splice switching  oligos (SSOs) to reverse this aberrant splicing in the pre-mRNA. With these lead  SSOs we show aberrant to wild type splice switching. This switching leads to an  increase of adult hemoglobin (HbA) up to 80\\% in erythroid cells from patients  with the IVS-2-745 mutation. Furthermore, we demonstrate a restoration of the  balance between β-like- and α-globin chains, and up to an 87\\% reduction in toxic  α-heme aggregates. While examining the potential benefit of 2'-MOE-SSOs in a  mixed sickle-thalassemic phenotypic setting, we found reduced HbS synthesis and  sickle cell formation due to HbA induction. In summary, 2'-MOE-SSOs are a  promising therapy for forms of β-thalassemia caused by mutations leading to  aberrant splicing.},\n\tlanguage = {eng},\n\tnumber = {5},\n\tjournal = {Haematologica},\n\tauthor = {Dong, Alisa and Ghiaccio, Valentina and Motta, Irene and Guo, Shuling and Peralta, Raechel and Freier, Susan M. and Watt, Andy and Damle, Sagar and Ikawa, Yasuhiro and Jarocha, Danuta and Chappell, Maxwell and Stephanou, Coralea and Delbini, Paola and Chen, Connie and Christou, Soteroula and Kleanthous, Marina and Smith-Whitley, Kim and Manwani, Deepa and Casu, Carla and Abdulmalik, Osheiza and Cappellini, Maria Domenica and Rivella, Stefano and Breda, Laura},\n\tmonth = may,\n\tyear = {2019},\n\tpmid = {32439726},\n\tpmcid = {PMC8094087},\n\tnote = {Place: Italy},\n\tpages = {1433--1442},\n}\n

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\n\n\n

\n β-thalassemia is a disorder caused by altered hemoglobin protein synthesis and affects individuals worldwide. Severe forms of the disease, left untreated, can result in death before the age of 3 years (1). The standard of care consists of chronic and costly palliative treatment by blood transfusion combined with iron chelation. This dual approach suppresses anemia and reduces iron-related toxicities in patients. Allogeneic bone marrow transplant is an option, but limited by the availability of a highly compatible HSC donor. While gene therapy is been explored in several trials, its use is highly limited to developed regions with centers of excellence and well-established healthcare systems (2). Hence, there remains a tremendous unmet medical need to develop alternative treatment strategies for β-thalassemia (3). Occurrence of aberrant splicing is one of the processes that affects β-globin synthesis in β-thalassemia. The (C\\textgreaterG) IVS-2-745 is a splicing mutation within intron 2 of the β-globin gene. It leads to an aberrantly spliced mRNA that incorporates an intron fragment. This results in an in-frame premature termination codon that inhibits β-globin production. Here, we propose the use of uniform 2'-O-methoxyethyl (2'-MOE) splice switching oligos (SSOs) to reverse this aberrant splicing in the pre-mRNA. With these lead SSOs we show aberrant to wild type splice switching. This switching leads to an increase of adult hemoglobin (HbA) up to 80% in erythroid cells from patients with the IVS-2-745 mutation. Furthermore, we demonstrate a restoration of the balance between β-like- and α-globin chains, and up to an 87% reduction in toxic α-heme aggregates. While examining the potential benefit of 2'-MOE-SSOs in a mixed sickle-thalassemic phenotypic setting, we found reduced HbS synthesis and sickle cell formation due to HbA induction. In summary, 2'-MOE-SSOs are a promising therapy for forms of β-thalassemia caused by mutations leading to aberrant splicing.\n

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\n \n 2018\n \n \n (7)\n \n \n

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\n \n\n \n \n \n \n \n First study on iron complexes in blood and organ samples from thalassaemic and normal laboratory mice using Mössbauer spectroscopy.\n \n \n \n\n\n \n Charitou, G., Petousis, V., Tsertos, C., Parpottas, Y., Kleanthous, M., Phylactides, M., & Christou, S.\n\n\n \n\n\n\n European biophysics journal : EBJ, 47(2): 131–138. March 2018.\n Place: Germany\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{charitou_first_2018,\n\ttitle = {First study on iron complexes in blood and organ samples from thalassaemic and normal laboratory mice using {Mössbauer} spectroscopy.},\n\tvolume = {47},\n\tissn = {1432-1017 0175-7571},\n\tdoi = {10.1007/s00249-017-1234-6},\n\tabstract = {Measurements of iron complexes and iron stores in the body are crucial for evaluation and management of chelation therapy targeted against iron accumulation  or overload in blood and organs. In this work, blood and tissue samples from one  normal and one thalassaemic laboratory mouse were studied using (57)Fe Mössbauer  spectroscopy at 78 K for the first time. In contrast to human patients, these  laboratory mice did not receive any medical treatment, thus the iron components  present in the samples are not altered from their natural state. The Mössbauer  spectra of blood, liver and spleen samples of the thalassaemic mouse were found  to differ in shape and iron content compared with corresponding spectra of the  normal mouse. These results demonstrate a basis for further exploitation of the  thalassaemic mouse model to study thalassaemia and its treatment in more detail  using Mössbauer spectroscopy.},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {European biophysics journal : EBJ},\n\tauthor = {Charitou, George and Petousis, Vlassis and Tsertos, Charalambos and Parpottas, Yannis and Kleanthous, Marina and Phylactides, Marios and Christou, Soteroula},\n\tmonth = mar,\n\tyear = {2018},\n\tpmid = {28695249},\n\tnote = {Place: Germany},\n\tkeywords = {*Spectroscopy, Mossbauer, Animals, Blood, Iron/blood/*chemistry/*metabolism, Liver, Liver/metabolism, Mice, Mice, Inbred C57BL, Mössbauer spectroscopy, Spleen, Spleen/metabolism, Thalassaemia, Thalassemia/*blood/*metabolism},\n\tpages = {131--138},\n}\n\n

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\n \n\n \n \n \n \n \n Fast Temperature-Gradient COLD PCR for the enrichment of the paternally inherited SNPs in cell free fetal DNA; an application to non-invasive prenatal diagnosis of β-thalassaemia.\n \n \n \n\n\n \n Byrou, S., Makrigiorgos, G. M., Christofides, A., Kallikas, I., Papasavva, T., & Kleanthous, M.\n\n\n \n\n\n\n PloS one, 13(7): e0200348. 2018.\n Place: United States\n\n\n\n
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@article{byrou_fast_2018,\n\ttitle = {Fast {Temperature}-{Gradient} {COLD} {PCR} for the enrichment of the paternally inherited {SNPs} in cell free fetal {DNA}; an application to non-invasive prenatal diagnosis of  β-thalassaemia.},\n\tvolume = {13},\n\tissn = {1932-6203},\n\tdoi = {10.1371/journal.pone.0200348},\n\tabstract = {OBJECTIVE: To develop a sensitive, specific, simple, cost-effective and reproducible platform for the non-invasive prenatal detection of paternally  inherited alleles for β-thalassaemia. The development of such an assay is of  major significance in order to replace currently-applied invasive methods  containing inherent fetal loss risks. METHODS: We present a fast  Temperature-Gradient Co-amplification at Lower Denaturation Temperature  Polymerase Chain Reaction (fast TG COLD PCR) methodology for the detection of the  paternally-inherited fetal alleles in maternal plasma. Two single-nucleotide  polymorphisms (SNPs), rs7480526 (G/T) and rs968857 (G/A) that are located on the  β-globin gene cluster and exhibit a high degree of heterozygosity in the Cypriot  population were selected for evaluation. Seventeen maternal plasma samples from  pregnancies at risk for β-thalassemia were analysed for the selected SNPs using  the novel fast TG COLD PCR assay. RESULTS: Using fast TG COLD PCR, the paternally  inherited allele in cell free fetal DNA was correctly determined for all the 17  maternal plasma samples tested, showing full agreement with the Chorionic Villus  Sampling (CVS) analysis. CONCLUSIONS: Our findings are encouraging and  demonstrate the efficiency and sensitivity of fast TG COLD PCR in detecting the  minor paternally-inherited fetal alleles in maternal plasma for the development  of a NIPD assay for β-thalassaemia.},\n\tlanguage = {eng},\n\tnumber = {7},\n\tjournal = {PloS one},\n\tauthor = {Byrou, Stefania and Makrigiorgos, G. Mike and Christofides, Agathoklis and Kallikas, Ioannis and Papasavva, Thessalia and Kleanthous, Marina},\n\tyear = {2018},\n\tpmid = {30044883},\n\tpmcid = {PMC6059429},\n\tnote = {Place: United States},\n\tkeywords = {*Paternal Inheritance, *Polymorphism, Single Nucleotide, Alleles, Cell-Free Nucleic Acids/*analysis/blood, Female, Gene Frequency, Genotyping Techniques, Humans, Polymerase Chain Reaction/*methods, Pregnancy, Prenatal Diagnosis/*methods, Sensitivity and Specificity, Temperature, beta-Thalassemia/*diagnosis/genetics},\n\tpages = {e0200348},\n}\n\n

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\n \n\n \n \n \n \n \n Corrigendum to \"European contribution to the study of ROS: A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS)\" [Redox Biol. 13 (2017) 94-162].\n \n \n \n\n\n \n Egea, J., Fabregat, I., Frapart, Y. M., Ghezzi, P., Görlach, A., Kietzmann, T., Kubaichuk, K., Knaus, U. G., Lopez, M. G., Olaso-Gonzalez, G., Petry, A., Schulz, R., Vina, J., Winyard, P., Abbas, K., Ademowo, O. S., Afonso, C. B., Andreadou, I., Antelmann, H., Antunes, F., Aslan, M., Bachschmid, M. M., Barbosa, R. M., Belousov, V., Berndt, C., Bernlohr, D., Bertrán, E., Bindoli, A., Bottari, S. P., Brito, P. M., Carrara, G., Casas, A. I., Chatzi, A., Chondrogianni, N., Conrad, M., Cooke, M. S., Costa, J. G., Cuadrado, A., My-Chan Dang, P., De Smet, B., Debelec-Butuner, B., Dias, I. H. K., Dunn, J. D., Edson, A. J., El Assar, M., El-Benna, J., Ferdinandy, P., Fernandes, A. S., Fladmark, K. E., Förstermann, U., Giniatullin, R., Giricz, Z., Görbe, A., Griffiths, H., Hampl, V., Hanf, A., Herget, J., Hernansanz-Agustín, P., Hillion, M., Huang, J., Ilikay, S., Jansen-Dürr, P., Jaquet, V., Joles, J. A., Kalyanaraman, B., Kaminskyy, D., Karbaschi, M., Kleanthous, M., Klotz, L. O., Korac, B., Korkmaz, K. S., Koziel, R., Kračun, D., Krause, K. H., Křen, V., Krieg, T., Laranjinha, J., Lazou, A., Li, H., Martínez-Ruiz, A., Matsui, R., McBean, G. J., Meredith, S. P., Messens, J., Miguel, V., Mikhed, Y., Milisav, I., Milković, L., Miranda-Vizuete, A., Mojović, M., Monsalve, M., Mouthuy, P. A., Mulvey, J., Münzel, T., Muzykantov, V., Nguyen, I. T. N., Oelze, M., Oliveira, N. G., Palmeira, C. M., Papaevgeniou, N., Pavićević, A., Pedre, B., Peyrot, F., Phylactides, M., Pircalabioru, G. G., Pitt, A. R., Poulsen, H. E., Prieto, I., Rigobello, M. P., Robledinos-Antón, N., Rodríguez-Mañas, L., Rolo, A. P., Rousset, F., Ruskovska, T., Saraiva, N., Sasson, S., Schröder, K., Semen, K., Seredenina, T., Shakirzyanova, A., Smith, G. L., Soldati, T., Sousa, B. C., Spickett, C. M., Stancic, A., Stasia, M. J., Steinbrenner, H., Stepanić, V., Steven, S., Tokatlidis, K., Tuncay, E., Turan, B., Ursini, F., Vacek, J., Vajnerova, O., Valentová, K., Van Breusegem, F., Varisli, L., Veal, E. A., Yalçın, A. S., Yelisyeyeva, O., Žarković, N., Zatloukalová, M., Zielonka, J., Touyz, R. M., Papapetropoulos, A., Grune, T., Lamas, S., Schmidt, H. H. H. W., Di Lisa, F., & Daiber, A.\n\n\n \n\n\n\n Redox biology, 14: 694–696. April 2018.\n Place: Netherlands\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{egea_corrigendum_2018,\n\ttitle = {Corrigendum to "{European} contribution to the study of {ROS}: {A} summary of the findings and prospects for the future from the {COST} action {BM1203} ({EU}-{ROS})"  [{Redox} {Biol}. 13 (2017) 94-162].},\n\tvolume = {14},\n\tissn = {2213-2317},\n\tdoi = {10.1016/j.redox.2017.10.001},\n\tlanguage = {eng},\n\tjournal = {Redox biology},\n\tauthor = {Egea, J. and Fabregat, I. and Frapart, Y. M. and Ghezzi, P. and Görlach, A. and Kietzmann, T. and Kubaichuk, K. and Knaus, U. G. and Lopez, M. G. and Olaso-Gonzalez, G. and Petry, A. and Schulz, R. and Vina, J. and Winyard, P. and Abbas, K. and Ademowo, O. S. and Afonso, C. B. and Andreadou, I. and Antelmann, H. and Antunes, F. and Aslan, M. and Bachschmid, M. M. and Barbosa, R. M. and Belousov, V. and Berndt, C. and Bernlohr, D. and Bertrán, E. and Bindoli, A. and Bottari, S. P. and Brito, P. M. and Carrara, G. and Casas, A. I. and Chatzi, A. and Chondrogianni, N. and Conrad, M. and Cooke, M. S. and Costa, J. G. and Cuadrado, A. and My-Chan Dang, P. and De Smet, B. and Debelec-Butuner, B. and Dias, I. H. K. and Dunn, J. D. and Edson, A. J. and El Assar, M. and El-Benna, J. and Ferdinandy, P. and Fernandes, A. S. and Fladmark, K. E. and Förstermann, U. and Giniatullin, R. and Giricz, Z. and Görbe, A. and Griffiths, H. and Hampl, V. and Hanf, A. and Herget, J. and Hernansanz-Agustín, P. and Hillion, M. and Huang, J. and Ilikay, S. and Jansen-Dürr, P. and Jaquet, V. and Joles, J. A. and Kalyanaraman, B. and Kaminskyy, D. and Karbaschi, M. and Kleanthous, M. and Klotz, L. O. and Korac, B. and Korkmaz, K. S. and Koziel, R. and Kračun, D. and Krause, K. H. and Křen, V. and Krieg, T. and Laranjinha, J. and Lazou, A. and Li, H. and Martínez-Ruiz, A. and Matsui, R. and McBean, G. J. and Meredith, S. P. and Messens, J. and Miguel, V. and Mikhed, Y. and Milisav, I. and Milković, L. and Miranda-Vizuete, A. and Mojović, M. and Monsalve, M. and Mouthuy, P. A. and Mulvey, J. and Münzel, T. and Muzykantov, V. and Nguyen, I. T. N. and Oelze, M. and Oliveira, N. G. and Palmeira, C. M. and Papaevgeniou, N. and Pavićević, A. and Pedre, B. and Peyrot, F. and Phylactides, M. and Pircalabioru, G. G. and Pitt, A. R. and Poulsen, H. E. and Prieto, I. and Rigobello, M. P. and Robledinos-Antón, N. and Rodríguez-Mañas, L. and Rolo, A. P. and Rousset, F. and Ruskovska, T. and Saraiva, N. and Sasson, S. and Schröder, K. and Semen, K. and Seredenina, T. and Shakirzyanova, A. and Smith, G. L. and Soldati, T. and Sousa, B. C. and Spickett, C. M. and Stancic, A. and Stasia, M. J. and Steinbrenner, H. and Stepanić, V. and Steven, S. and Tokatlidis, K. and Tuncay, E. and Turan, B. and Ursini, F. and Vacek, J. and Vajnerova, O. and Valentová, K. and Van Breusegem, F. and Varisli, L. and Veal, E. A. and Yalçın, A. S. and Yelisyeyeva, O. and Žarković, N. and Zatloukalová, M. and Zielonka, J. and Touyz, R. M. and Papapetropoulos, A. and Grune, T. and Lamas, S. and Schmidt, H. H. H. W. and Di Lisa, F. and Daiber, A.},\n\tmonth = apr,\n\tyear = {2018},\n\tpmid = {29107648},\n\tpmcid = {PMC5975209},\n\tnote = {Place: Netherlands},\n\tpages = {694--696},\n}\n\n

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\n \n\n \n \n \n \n \n Multiple endocrine neoplasia 2 in Cyprus: evidence for a founder effect.\n \n \n \n\n\n \n Fanis, P., Skordis, N., Frangos, S., Christopoulos, G., Spanou-Aristidou, E., Andreou, E., Manoli, P., Mavrommatis, M., Nicolaou, S., Kleanthous, M., Cariolou, M. A., Christophidou-Anastasiadou, V., Tanteles, G. A., Phylactou, L. A., & Neocleous, V.\n\n\n \n\n\n\n Journal of endocrinological investigation, 41(10): 1149–1157. October 2018.\n Place: Italy\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{fanis_multiple_2018,\n\ttitle = {Multiple endocrine neoplasia 2 in {Cyprus}: evidence for a founder effect.},\n\tvolume = {41},\n\tissn = {1720-8386 0391-4097},\n\tdoi = {10.1007/s40618-018-0841-0},\n\tabstract = {PURPOSE: Multiple endocrine neoplasia type 2 (MEN2) affects patients with RET proto-oncogene mutations. This cohort study refers to patients who were diagnosed  with familial medullary thyroid carcinoma (MTC) and underwent RET genetic testing  in Cyprus between years 2002 and 2017. METHODS AND PATIENTS: Forty patients  underwent RET testing by Sanger sequencing of exons 10-11 and 13-16. Genotyping  with STR genetic markers flanking the RET gene along with Y-chromosome genotyping  and haplogroup assignment was also performed. RESULTS: RET mutations were  identified in 40 patients from 11 apparently unrelated Cypriot families and two  non-familial sporadic cases. Nine probands (69.2\\%) were heterozygous for  p.Cys618Arg, one (7.7\\%) for p.Cys634Phe, one (7.7\\%) for the somatic delE632-L633  and two (15.4\\%) for p.Met918Thr mutations. The mean age at MTC diagnosis of  patients carrying p.Cys618Arg was 36.8 ± 14.2 years. The age of pheo diagnosis  ranged from 26 to 43 years and appeared simultaneously with MTC in 5/36 (13.9\\%)  cases. The high frequency of the p.Cys618Arg mutation suggested a possible  ancestral mutational event. Haplotype analysis was performed in families with and  without p.Cys618Arg. Six microsatellite markers covering the RET gene and  neighboring regions identified one core haplotype associated with all patients  carrying p.Cys618Arg mutation. CONCLUSIONS: The mutation p.Cys618Arg is by far  the most prevalent mutation in Cyprus followed by other reported mutations of  variable clinical significance. The provided molecular evidence speculates  p.Cys618Arg mutation as an ancestral mutation that has spread in Cyprus due to a  possible founder effect.},\n\tlanguage = {eng},\n\tnumber = {10},\n\tjournal = {Journal of endocrinological investigation},\n\tauthor = {Fanis, P. and Skordis, N. and Frangos, S. and Christopoulos, G. and Spanou-Aristidou, E. and Andreou, E. and Manoli, P. and Mavrommatis, M. and Nicolaou, S. and Kleanthous, M. and Cariolou, M. A. and Christophidou-Anastasiadou, V. and Tanteles, G. A. and Phylactou, L. A. and Neocleous, V.},\n\tmonth = oct,\n\tyear = {2018},\n\tpmid = {29396759},\n\tpmcid = {PMC6182349},\n\tnote = {Place: Italy},\n\tkeywords = {*Founder Effect, Adult, Arginine/genetics, Cancer, Carcinoma, Medullary/*congenital/diagnosis/epidemiology/genetics, Cohort Studies, Cyprus/epidemiology, Cysteine/genetics, Female, Humans, Male, Medullary thyroid carcinoma, Middle Aged, Multiple Endocrine Neoplasia Type 2a/diagnosis/*epidemiology/*genetics, Multiple endocrine neoplasia type 2, Pedigree, Pheochromocytoma, Proto-Oncogene Mas, Proto-Oncogene Proteins c-ret/*genetics, RET proto-oncogene, Thyroid Neoplasms/diagnosis/*epidemiology/*genetics},\n\tpages = {1149--1157},\n}\n\n

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\n \n\n \n \n \n \n \n Newborn screening for sickle cell disease in Europe: recommendations from a Pan-European Consensus Conference.\n \n \n \n\n\n \n Lobitz, S., Telfer, P., Cela, E., Allaf, B., Angastiniotis, M., Backman Johansson, C., Badens, C., Bento, C., Bouva, M. J., Canatan, D., Charlton, M., Coppinger, C., Daniel, Y., de Montalembert, M., Ducoroy, P., Dulin, E., Fingerhut, R., Frömmel, C., García-Morin, M., Gulbis, B., Holtkamp, U., Inusa, B., James, J., Kleanthous, M., Klein, J., Kunz, J. B., Langabeer, L., Lapouméroulie, C., Marcao, A., Marín Soria, J. L., McMahon, C., Ohene-Frempong, K., Périni, J., Piel, F. B., Russo, G., Sainati, L., Schmugge, M., Streetly, A., Tshilolo, L., Turner, C., Venturelli, D., Vilarinho, L., Yahyaoui, R., Elion, J., & Colombatti, R.\n\n\n \n\n\n\n British journal of haematology, 183(4): 648–660. November 2018.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{lobitz_newborn_2018,\n\ttitle = {Newborn screening for sickle cell disease in {Europe}: recommendations from a {Pan}-{European} {Consensus} {Conference}.},\n\tvolume = {183},\n\tcopyright = {© 2018 British Society for Haematology and John Wiley \\& Sons Ltd.},\n\tissn = {1365-2141 0007-1048},\n\tdoi = {10.1111/bjh.15600},\n\tabstract = {Sickle Cell Disease (SCD) is an increasing global health problem and presents significant challenges to European health care systems. Newborn screening (NBS)  for SCD enables early initiation of preventive measures and has contributed to a  reduction in childhood mortality from SCD. Policies and methodologies for NBS  vary in different countries, and this might have consequences for the quality of  care and clinical outcomes for SCD across Europe. A two-day Pan-European  consensus conference was held in Berlin in April 2017 in order to appraise the  current status of NBS for SCD and to develop consensus-based statements on  indications and methodology for NBS for SCD in Europe. More than 50 SCD experts  from 13 European countries participated in the conference. This paper aims to  summarise the discussions and present consensus recommendations which can be used  to support the development of NBS programmes in European countries where they do  not yet exist, and to review existing programmes.},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {British journal of haematology},\n\tauthor = {Lobitz, Stephan and Telfer, Paul and Cela, Elena and Allaf, Bichr and Angastiniotis, Michael and Backman Johansson, Carolina and Badens, Catherine and Bento, Celeste and Bouva, Marelle J. and Canatan, Duran and Charlton, Matthew and Coppinger, Cathy and Daniel, Yvonne and de Montalembert, Marianne and Ducoroy, Patrick and Dulin, Elena and Fingerhut, Ralph and Frömmel, Claudia and García-Morin, Marina and Gulbis, Béatrice and Holtkamp, Ute and Inusa, Baba and James, John and Kleanthous, Marina and Klein, Jeannette and Kunz, Joachim B. and Langabeer, Lisa and Lapouméroulie, Claudine and Marcao, Ana and Marín Soria, José L. and McMahon, Corrina and Ohene-Frempong, Kwaku and Périni, Jean-Marc and Piel, Frédéric B. and Russo, Giovanna and Sainati, Laura and Schmugge, Markus and Streetly, Allison and Tshilolo, Leon and Turner, Charles and Venturelli, Donatella and Vilarinho, Laura and Yahyaoui, Rachel and Elion, Jacques and Colombatti, Raffaella},\n\tmonth = nov,\n\tyear = {2018},\n\tpmid = {30334577},\n\tnote = {Place: England},\n\tkeywords = {Anemia, Sickle Cell/*diagnostic imaging/epidemiology/*genetics, Consensus Development Conferences as Topic, Europe/epidemiology, Female, Humans, Infant, Newborn, Male, Neonatal Screening, Practice Guidelines as Topic, haemoglobinopathies, newborn screening, prevention, sickle cell anaemia, sickle cell disease},\n\tpages = {648--660},\n}\n\n

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\n \n\n \n \n \n \n \n Rapid and Sensitive Assessment of Globin Chains for Gene and Cell Therapy of Hemoglobinopathies.\n \n \n \n\n\n \n Loucari, C. C., Patsali, P., van Dijk, T. B., Stephanou, C., Papasavva, P., Zanti, M., Kurita, R., Nakamura, Y., Christou, S., Sitarou, M., Philipsen, S., Lederer, C. W., & Kleanthous, M.\n\n\n \n\n\n\n Human gene therapy methods, 29(1): 60–74. February 2018.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{loucari_rapid_2018,\n\ttitle = {Rapid and {Sensitive} {Assessment} of {Globin} {Chains} for {Gene} and {Cell} {Therapy} of {Hemoglobinopathies}.},\n\tvolume = {29},\n\tissn = {1946-6544 1946-6536},\n\tdoi = {10.1089/hgtb.2017.190},\n\tabstract = {The β-hemoglobinopathies sickle cell anemia and β-thalassemia are the focus of many gene-therapy studies. A key disease parameter is the abundance of globin  chains because it indicates the level of anemia, likely toxicity of excess or  aberrant globins, and therapeutic potential of induced or exogenous β-like  globins. Reversed-phase high-performance liquid chromatography (HPLC) allows  versatile and inexpensive globin quantification, but commonly applied protocols  suffer from long run times, high sample requirements, or inability to separate  murine from human β-globin chains. The latter point is problematic for in vivo  studies with gene-addition vectors in murine disease models and mouse/human  chimeras. This study demonstrates HPLC-based measurements of globin expression  (1) after differentiation of the commonly applied human umbilical cord  blood-derived erythroid progenitor-2 cell line, (2) in erythroid progeny of  CD34(+) cells for the analysis of clustered regularly interspaced short  palindromic repeats/Cas9-mediated disruption of the globin regulator BCL11A, and  (3) of transgenic mice holding the human β-globin locus. At run times of 8 min  for separation of murine and human β-globin chains as well as of human γ-globin  chains, and with routine measurement of globin-chain ratios for 12 nL of blood  (tested for down to 0.75 nL) or of 300,000 in vitro differentiated cells, the  methods presented here and any variant-specific adaptations thereof will greatly  facilitate evaluation of novel therapy applications for β-hemoglobinopathies.},\n\tlanguage = {eng},\n\tnumber = {1},\n\tjournal = {Human gene therapy methods},\n\tauthor = {Loucari, Constantinos C. and Patsali, Petros and van Dijk, Thamar B. and Stephanou, Coralea and Papasavva, Panayiota and Zanti, Maria and Kurita, Ryo and Nakamura, Yukio and Christou, Soteroulla and Sitarou, Maria and Philipsen, Sjaak and Lederer, Carsten W. and Kleanthous, Marina},\n\tmonth = feb,\n\tyear = {2018},\n\tpmid = {29325430},\n\tpmcid = {PMC5806072},\n\tnote = {Place: United States},\n\tkeywords = {*Anemia, Sickle Cell/genetics/metabolism/pathology/therapy, *Genetic Vectors/genetics/metabolism, *beta-Globins/biosynthesis/genetics, *gamma-Globins/genetics, Animals, CRISPR/Cas9, Cell Line, Cell- and Tissue-Based Therapy/*methods, Disease Models, Animal, Genetic Therapy/*methods, Humans, Mice, Mice, Transgenic, fetal hemoglobin, gene addition, high-performance liquid chromatography, thalassemia, β-hemoglobinopathy},\n\tpages = {60--74},\n}\n\n

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\n \n\n \n \n \n \n \n Selection and Identification of Skeletal-Muscle-Targeted RNA Aptamers.\n \n \n \n\n\n \n Philippou, S., Mastroyiannopoulos, N. P., Makrides, N., Lederer, C. W., Kleanthous, M., & Phylactou, L. A.\n\n\n \n\n\n\n Molecular therapy. Nucleic acids, 10: 199–214. March 2018.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{philippou_selection_2018,\n\ttitle = {Selection and {Identification} of {Skeletal}-{Muscle}-{Targeted} {RNA} {Aptamers}.},\n\tvolume = {10},\n\tcopyright = {Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.},\n\tissn = {2162-2531},\n\tdoi = {10.1016/j.omtn.2017.12.004},\n\tabstract = {Oligonucleotide gene therapy has shown great promise for the treatment of muscular dystrophies. Nevertheless, the selective delivery to affected muscles  has shown to be challenging because of their high representation in the body and  the high complexity of their cell membranes. Current trials show loss of  therapeutic molecules to non-target tissues leading to lower target efficacy.  Therefore, strategies that increase uptake efficiency would be particularly  compelling. To address this need, we applied a cell-internalization SELEX  (Systematic Evolution of Ligands by Exponential Enrichment) approach and  identified a skeletal muscle-specific RNA aptamer. A01B RNA aptamer  preferentially internalizes in skeletal muscle cells and exhibits decreased  affinity for off-target cells. Moreover, this in vitro selected aptamer retained  its functionality in vivo, suggesting a potential new approach for targeting  skeletal muscles. Ultimately, this will aid in the development of targeted  oligonucleotide therapies against muscular dystrophies.},\n\tlanguage = {eng},\n\tjournal = {Molecular therapy. Nucleic acids},\n\tauthor = {Philippou, Styliana and Mastroyiannopoulos, Nikolaos P. and Makrides, Neoklis and Lederer, Carsten W. and Kleanthous, Marina and Phylactou, Leonidas A.},\n\tmonth = mar,\n\tyear = {2018},\n\tpmid = {29499933},\n\tpmcid = {PMC5862129},\n\tnote = {Place: United States},\n\tkeywords = {SELEX, antisense therapy, aptamer, delivery, gene therapy, muscular dystrophies, skeletal muscle},\n\tpages = {199--214},\n}\n\n

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\n \n\n \n \n \n \n \n \n ITHANET: Information and database community portal for haemoglobinopathies.\n \n \n \n \n\n\n \n Kountouris, P., Stephanou, C., Bento, C., Fanis, P., Elion, J., Ramesar, R. S., Zilfalil, B. A., Robinson, H. M., Traeger-Synodinos, J., Human Variome Project Global Globin 2020 Challenge, Lederer, C. W., & Kleanthous, M.\n\n\n \n\n\n\n bioRxiv,209361. October 2017.\n \n\n\n\n
\n\n\n\n \n \n $\"ITHANET:$ Paper\n \n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{kountouris_ithanet_2017,\n\ttitle = {{ITHANET}: {Information} and database community portal for haemoglobinopathies},\n\tcopyright = {© 2017, Posted by Cold Spring Harbor Laboratory. This pre-print is available under a Creative Commons License (Attribution-NonCommercial-NoDerivs 4.0 International), CC BY-NC-ND 4.0, as described at http://creativecommons.org/licenses/by-nc-nd/4.0/},\n\tshorttitle = {{ITHANET}},\n\turl = {https://www.biorxiv.org/content/10.1101/209361v1},\n\tdoi = {10.1101/209361},\n\tabstract = {{\\textless}p{\\textgreater}Haemoglobinopathies are the commonest monogenic diseases, with millions of carriers and patients worldwide. Online resources for haemoglobinopathies are largely divided into specialised sites catering for patients, researchers and clinicians separately. However, the severity, ubiquity and surprising genetic complexity of the haemoglobinopathies call for an integrated website to serve as a free and comprehensive repository and tool for patients, scientists and health professionals alike. This paper presents the ITHANET community portal, an expanding resource for clinicians and researchers dealing with haemoglobinopathies. It integrates information on news, events, publications, clinical trials and haemoglobinopathy-related organisations and experts and, most importantly, databases of variations, epidemiology and diagnostic and clinical data. Specifically, ITHANET provides annotation for 2690 haemoglobinopathy-related variations, epidemiological data for more than 180 countries and information for more than 600 HPLC diagnostic reports. The ITHANET portal accepts and incorporates contributions to its content by local experts from any country in the world and is freely available for the public at http://www.ithanet.eu.{\\textless}/p{\\textgreater}},\n\tlanguage = {en},\n\turldate = {2019-01-24},\n\tjournal = {bioRxiv},\n\tauthor = {Kountouris, Petros and Stephanou, Coralea and Bento, Celeste and Fanis, Pavlos and Elion, Jacques and Ramesar, Raj S. and Zilfalil, Bin Alwi and Robinson, Helen M. and Traeger-Synodinos, Joanne and {Human Variome Project Global Globin 2020 Challenge} and Lederer, Carsten Werner and Kleanthous, Marina},\n\tmonth = oct,\n\tyear = {2017},\n\tpages = {209361},\n}\n\n

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\n \n\n \n \n \n \n \n European contribution to the study of ROS: A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS).\n \n \n \n\n\n \n Egea, J., Fabregat, I., Frapart, Y. M., Ghezzi, P., Görlach, A., Kietzmann, T., Kubaichuk, K., Knaus, U. G., Lopez, M. G., Olaso-Gonzalez, G., Petry, A., Schulz, R., Vina, J., Winyard, P., Abbas, K., Ademowo, O. S., Afonso, C. B., Andreadou, I., Antelmann, H., Antunes, F., Aslan, M., Bachschmid, M. M., Barbosa, R. M., Belousov, V., Berndt, C., Bernlohr, D., Bertrán, E., Bindoli, A., Bottari, S. P., Brito, P. M., Carrara, G., Casas, A. I., Chatzi, A., Chondrogianni, N., Conrad, M., Cooke, M. S., Costa, J. G., Cuadrado, A., My-Chan Dang, P., De Smet, B., Debelec-Butuner, B., Dias, I. H. K., Dunn, J. D., Edson, A. J., El Assar, M., El-Benna, J., Ferdinandy, P., Fernandes, A. S., Fladmark, K. E., Förstermann, U., Giniatullin, R., Giricz, Z., Görbe, A., Griffiths, H., Hampl, V., Hanf, A., Herget, J., Hernansanz-Agustín, P., Hillion, M., Huang, J., Ilikay, S., Jansen-Dürr, P., Jaquet, V., Joles, J. A., Kalyanaraman, B., Kaminskyy, D., Karbaschi, M., Kleanthous, M., Klotz, L., Korac, B., Korkmaz, K. S., Koziel, R., Kračun, D., Krause, K., Křen, V., Krieg, T., Laranjinha, J., Lazou, A., Li, H., Martínez-Ruiz, A., Matsui, R., McBean, G. J., Meredith, S. P., Messens, J., Miguel, V., Mikhed, Y., Milisav, I., Milković, L., Miranda-Vizuete, A., Mojović, M., Monsalve, M., Mouthuy, P., Mulvey, J., Münzel, T., Muzykantov, V., Nguyen, I. T. N., Oelze, M., Oliveira, N. G., Palmeira, C. M., Papaevgeniou, N., Pavićević, A., Pedre, B., Peyrot, F., Phylactides, M., Pircalabioru, G. G., Pitt, A. R., Poulsen, H. E., Prieto, I., Rigobello, M. P., Robledinos-Antón, N., Rodríguez-Mañas, L., Rolo, A. P., Rousset, F., Ruskovska, T., Saraiva, N., Sasson, S., Schröder, K., Semen, K., Seredenina, T., Shakirzyanova, A., Smith, G. L., Soldati, T., Sousa, B. C., Spickett, C. M., Stancic, A., Stasia, M. J., Steinbrenner, H., Stepanić, V., Steven, S., Tokatlidis, K., Tuncay, E., Turan, B., Ursini, F., Vacek, J., Vajnerova, O., Valentová, K., Van Breusegem, F., Varisli, L., Veal, E. A., Yalçın, A. S., Yelisyeyeva, O., Žarković, N., Zatloukalová, M., Zielonka, J., Touyz, R. M., Papapetropoulos, A., Grune, T., Lamas, S., Schmidt, H. H. H. W., Di Lisa, F., & Daiber, A.\n\n\n \n\n\n\n Redox biology, 13: 94–162. October 2017.\n Place: Netherlands\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{egea_european_2017,\n\ttitle = {European contribution to the study of {ROS}: {A} summary of the findings and prospects for the future from the {COST} action {BM1203} ({EU}-{ROS}).},\n\tvolume = {13},\n\tcopyright = {Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.},\n\tissn = {2213-2317},\n\tdoi = {10.1016/j.redox.2017.05.007},\n\tabstract = {The European Cooperation in Science and Technology (COST) provides an ideal framework to establish multi-disciplinary research networks. COST Action BM1203 (EU-ROS) represents a consortium of researchers from different disciplines who are dedicated to providing new insights and tools for better understanding redox biology and medicine and, in the long run, to finding new therapeutic strategies to target dysregulated redox processes in various diseases. This report highlights the major achievements of EU-ROS as well as research updates and new perspectives arising from its members. The EU-ROS consortium comprised more than 140 active members who worked together for four years on the topics briefly described below. The formation of reactive oxygen and nitrogen species (RONS) is an established hallmark of our aerobic environment and metabolism but RONS also act as messengers via redox regulation of essential cellular processes. The fact that many diseases have been found to be associated with oxidative stress established the theory of oxidative stress as a trigger of diseases that can be corrected by antioxidant therapy. However, while experimental studies support this thesis, clinical studies still generate controversial results, due to complex pathophysiology of oxidative stress in humans. For future improvement of antioxidant therapy and better understanding of redox-associated disease progression detailed knowledge on the sources and targets of RONS formation and discrimination of their detrimental or beneficial roles is required. In order to advance this important area of biology and medicine, highly synergistic approaches combining a variety of diverse and contrasting disciplines are needed.},\n\tlanguage = {eng},\n\tjournal = {Redox biology},\n\tauthor = {Egea, Javier and Fabregat, Isabel and Frapart, Yves M. and Ghezzi, Pietro and Görlach, Agnes and Kietzmann, Thomas and Kubaichuk, Kateryna and Knaus, Ulla G. and Lopez, Manuela G. and Olaso-Gonzalez, Gloria and Petry, Andreas and Schulz, Rainer and Vina, Jose and Winyard, Paul and Abbas, Kahina and Ademowo, Opeyemi S. and Afonso, Catarina B. and Andreadou, Ioanna and Antelmann, Haike and Antunes, Fernando and Aslan, Mutay and Bachschmid, Markus M. and Barbosa, Rui M. and Belousov, Vsevolod and Berndt, Carsten and Bernlohr, David and Bertrán, Esther and Bindoli, Alberto and Bottari, Serge P. and Brito, Paula M. and Carrara, Guia and Casas, Ana I. and Chatzi, Afroditi and Chondrogianni, Niki and Conrad, Marcus and Cooke, Marcus S. and Costa, João G. and Cuadrado, Antonio and My-Chan Dang, Pham and De Smet, Barbara and Debelec-Butuner, Bilge and Dias, Irundika H. K. and Dunn, Joe Dan and Edson, Amanda J. and El Assar, Mariam and El-Benna, Jamel and Ferdinandy, Péter and Fernandes, Ana S. and Fladmark, Kari E. and Förstermann, Ulrich and Giniatullin, Rashid and Giricz, Zoltán and Görbe, Anikó and Griffiths, Helen and Hampl, Vaclav and Hanf, Alina and Herget, Jan and Hernansanz-Agustín, Pablo and Hillion, Melanie and Huang, Jingjing and Ilikay, Serap and Jansen-Dürr, Pidder and Jaquet, Vincent and Joles, Jaap A. and Kalyanaraman, Balaraman and Kaminskyy, Danylo and Karbaschi, Mahsa and Kleanthous, Marina and Klotz, Lars-Oliver and Korac, Bato and Korkmaz, Kemal Sami and Koziel, Rafal and Kračun, Damir and Krause, Karl-Heinz and Křen, Vladimír and Krieg, Thomas and Laranjinha, João and Lazou, Antigone and Li, Huige and Martínez-Ruiz, Antonio and Matsui, Reiko and McBean, Gethin J. and Meredith, Stuart P. and Messens, Joris and Miguel, Verónica and Mikhed, Yuliya and Milisav, Irina and Milković, Lidija and Miranda-Vizuete, Antonio and Mojović, Miloš and Monsalve, María and Mouthuy, Pierre-Alexis and Mulvey, John and Münzel, Thomas and Muzykantov, Vladimir and Nguyen, Isabel T. N. and Oelze, Matthias and Oliveira, Nuno G. and Palmeira, Carlos M. and Papaevgeniou, Nikoletta and Pavićević, Aleksandra and Pedre, Brandán and Peyrot, Fabienne and Phylactides, Marios and Pircalabioru, Gratiela G. and Pitt, Andrew R. and Poulsen, Henrik E. and Prieto, Ignacio and Rigobello, Maria Pia and Robledinos-Antón, Natalia and Rodríguez-Mañas, Leocadio and Rolo, Anabela P. and Rousset, Francis and Ruskovska, Tatjana and Saraiva, Nuno and Sasson, Shlomo and Schröder, Katrin and Semen, Khrystyna and Seredenina, Tamara and Shakirzyanova, Anastasia and Smith, Geoffrey L. and Soldati, Thierry and Sousa, Bebiana C. and Spickett, Corinne M. and Stancic, Ana and Stasia, Marie José and Steinbrenner, Holger and Stepanić, Višnja and Steven, Sebastian and Tokatlidis, Kostas and Tuncay, Erkan and Turan, Belma and Ursini, Fulvio and Vacek, Jan and Vajnerova, Olga and Valentová, Kateřina and Van Breusegem, Frank and Varisli, Lokman and Veal, Elizabeth A. and Yalçın, A. Suha and Yelisyeyeva, Olha and Žarković, Neven and Zatloukalová, Martina and Zielonka, Jacek and Touyz, Rhian M. and Papapetropoulos, Andreas and Grune, Tilman and Lamas, Santiago and Schmidt, Harald H. H. W. and Di Lisa, Fabio and Daiber, Andreas},\n\tmonth = oct,\n\tyear = {2017},\n\tpmid = {28577489},\n\tpmcid = {PMC5458069},\n\tnote = {Place: Netherlands},\n\tkeywords = {*International Cooperation, Animals, Antioxidants, European Union, Humans, Molecular Biology/organization \\& administration/trends, Oxidation-Reduction, Oxidative stress, Reactive Oxygen Species/chemistry/*metabolism, Reactive nitrogen species, Reactive oxygen species, Redox signaling, Redox therapeutics, Signal Transduction, Societies, Scientific},\n\tpages = {94--162},\n}\n\n

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\n \n\n \n \n \n \n \n Development of a Predictive Pharmacophore Model and a 3D-QSAR Study for an in silico Screening of New Potent Bcr-Abl Kinase Inhibitors.\n \n \n \n\n\n \n Vrontaki, E., Melagraki, G., Voskou, S., Phylactides, M. S., Mavromoustakos, T., Kleanthous, M., & Afantitis, A.\n\n\n \n\n\n\n Mini reviews in medicinal chemistry, 17(3): 188–204. 2017.\n Place: Netherlands\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{vrontaki_development_2017,\n\ttitle = {Development of a {Predictive} {Pharmacophore} {Model} and a {3D}-{QSAR} {Study} for an in silico {Screening} of {New} {Potent} {Bcr}-{Abl} {Kinase} {Inhibitors}.},\n\tvolume = {17},\n\tissn = {1875-5607 1389-5575},\n\tdoi = {10.2174/1389557516999160629101709},\n\tabstract = {Chronic myelogenous leukemia (CML) is a myeloproliferative disorder, characterized, in most cases, by the presence of the Bcr-Abl fusion oncogene.  Bcr-Abl is a constitutively active tyrosine kinase that is responsible for the  malignant transformation. Targeting the Bcr-Abl kinase is an attractive treatment  strategy for CML. First and second generation Bcr-Abl inhibitors have focused on  targeting the ATP-binding domain of the kinase. Mutations in that region are  relatively resistant to drug manipulation. Therefore, non-ATP-competitive agents  have been recently developed and tested. In the present study, in an attempt to  aid the design of new chemotypes with enhanced cytotoxicity against K562 cells,  3D pharmacophore models were generated and 3D-QSAR CoMFA and CoMSIA studies were  carried out on the 33 novel Abl kinase inhibitors (E)-\\&\\#945;-benzylthio chalcones  synthesized by Reddy et al. A five-point pharmacophore with a hydrogen bond  acceptor, two hydrophobic groups and two aromatic rings as pharmacophore  features, and a statistically significant 3D-QSAR model with excellent predictive  power were developed. The pharmacophore model was also used for alignment of the  33 compounds in a CoMFA/CoMSIA analysis. The contour maps of the fields of CoMFA  and CoMSIA models were utilized to provide structural insight into how these  molecules promote their toxicity. The possibility of using this model for the  design of drugs for the treatment of \\&\\#946;-thalassemia and sickle cell disease  (SCD), since several Bcr-Abl inhibitors are able to promote erythroid  differentiation and \\&\\#947;-globin expression in CML cell lines and primary  erythroid cells is discussed.},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {Mini reviews in medicinal chemistry},\n\tauthor = {Vrontaki, Eleni and Melagraki, Georgia and Voskou, Stella and Phylactides, Marios S. and Mavromoustakos, Thomas and Kleanthous, Marina and Afantitis, Antreas},\n\tyear = {2017},\n\tpmid = {28143387},\n\tnote = {Place: Netherlands},\n\tkeywords = {*Computer Simulation, *Drug Evaluation, Preclinical, *Models, Molecular, *Quantitative Structure-Activity Relationship, Antineoplastic Agents/chemistry/*pharmacology, Fusion Proteins, bcr-abl/*antagonists \\& inhibitors/metabolism, Humans, Leukemia, Myelogenous, Chronic, BCR-ABL Positive/*drug therapy/metabolism, Molecular Structure, Protein Kinase Inhibitors/chemistry/*pharmacology},\n\tpages = {188--204},\n}\n\n

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\n \n\n \n \n \n \n \n Suitability of small diagnostic peripheral-blood samples for cell-therapy studies.\n \n \n \n\n\n \n Stephanou, C., Papasavva, P., Zachariou, M., Patsali, P., Epitropou, M., Ladas, P., Al-Abdulla, R., Christou, S., Antoniou, M. N., Lederer, C. W., & Kleanthous, M.\n\n\n \n\n\n\n Cytotherapy, 19(2): 311–326. February 2017.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{stephanou_suitability_2017,\n\ttitle = {Suitability of small diagnostic peripheral-blood samples for cell-therapy studies.},\n\tvolume = {19},\n\tcopyright = {Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.},\n\tissn = {1477-2566 1465-3249},\n\tdoi = {10.1016/j.jcyt.2016.11.007},\n\tabstract = {BACKGROUND AIMS: Primary hematopoietic stem and progenitor cells (HSPCs) are key components of cell-based therapies for blood disorders and are thus the authentic  substrate for related research. We propose that ubiquitous small-volume  diagnostic samples represent a readily available and as yet untapped resource of  primary patient-derived cells for cell- and gene-therapy studies. METHODS: In the  present study we compare isolation and storage methods for HSPCs from normal and  thalassemic small-volume blood samples, considering genotype, density-gradient  versus lysis-based cell isolation and cryostorage media with different serum  contents. Downstream analyses include viability, recovery, differentiation in  semi-solid media and performance in liquid cultures and viral transductions.  RESULTS: We demonstrate that HSPCs isolated either by ammonium-chloride potassium  (ACK)-based lysis or by gradient isolation are suitable for functional analyses  in clonogenic assays, high-level HSPC expansion and efficient lentiviral  transduction. For cryostorage of cells, gradient isolation is superior to ACK  lysis, and cryostorage in freezing media containing 50\\% fetal bovine serum  demonstrated good results across all tested criteria. For assays on freshly  isolated cells, ACK lysis performed similar to, and for thalassemic samples  better than, gradient isolation, at a fraction of the cost and hands-on time. All  isolation and storage methods show considerable variation within sample groups,  but this is particularly acute for density gradient isolation of thalassemic  samples. DISCUSSION: This study demonstrates the suitability of small-volume  blood samples for storage and preclinical studies, opening up the research field  of HSPC and gene therapy to any blood diagnostic laboratory with corresponding  bioethics approval for experimental use of surplus material.},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {Cytotherapy},\n\tauthor = {Stephanou, Coralea and Papasavva, Panayiota and Zachariou, Myria and Patsali, Petros and Epitropou, Marilena and Ladas, Petros and Al-Abdulla, Ruba and Christou, Soteroulla and Antoniou, Michael N. and Lederer, Carsten W. and Kleanthous, Marina},\n\tmonth = feb,\n\tyear = {2017},\n\tpmid = {28088294},\n\tnote = {Place: England},\n\tkeywords = {Blood Preservation/methods/standards, Blood Specimen Collection/*methods/*standards, Cell Proliferation, Cell Separation/*methods/*standards, Cell Survival, Cell- and Tissue-Based Therapy/*methods, Cells, Cultured, Cryopreservation, Feasibility Studies, Freezing, Hematopoietic Stem Cells/pathology/physiology, Humans, Leukocyte Count, Leukocytes/*pathology/physiology, Serologic Tests, Thalassemia/*blood/pathology, colony-forming cell assay, cryopreservation, density-gradient separation, erythrocyte lysis by ACK buffer, hematopoietic stem and progenitor cell, in vitro expansion, lentiviral transduction, β-thalassemia},\n\tpages = {311--326},\n}\n\n

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\n \n 2016\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n A Tribute to George Stamatoyannopoulos.\n \n \n \n\n\n \n Srivastava, A., Kay, M. A., Athanasopoulos, T., Angastiniotis, M., Anagnostopoulos, A., Karponi, G., Yannaki, E., Zon, L. I., Lederer, C. W., Phylactides, M. S., & Kleanthous, M.\n\n\n \n\n\n\n Human gene therapy, 27(4): 280–286. April 2016.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{srivastava_tribute_2016,\n\ttitle = {A {Tribute} to {George} {Stamatoyannopoulos}.},\n\tvolume = {27},\n\tissn = {1557-7422 1043-0342},\n\tdoi = {10.1089/hum.2016.29025.gst},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {Human gene therapy},\n\tauthor = {Srivastava, Arun and Kay, Mark A. and Athanasopoulos, Takis and Angastiniotis, Michael and Anagnostopoulos, Achilles and Karponi, Garyfalia and Yannaki, Evangelia and Zon, Leonard I. and Lederer, Carsten W. and Phylactides, Marios S. and Kleanthous, Marina},\n\tmonth = apr,\n\tyear = {2016},\n\tpmid = {27074151},\n\tnote = {Place: United States},\n\tkeywords = {Animals, Dependovirus/genetics, Genetic Therapy/*history, History, 20th Century, History, 21st Century, Humans},\n\tpages = {280--286},\n}\n\n

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\n \n\n \n \n \n \n \n Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative PCR.\n \n \n \n\n\n \n Christodoulou, I., Patsali, P., Stephanou, C., Antoniou, M., Kleanthous, M., & Lederer, C. W.\n\n\n \n\n\n\n Gene therapy, 23(1): 113–118. January 2016.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{christodoulou_measurement_2016,\n\ttitle = {Measurement of lentiviral vector titre and copy number by cross-species duplex quantitative {PCR}.},\n\tvolume = {23},\n\tissn = {1476-5462 0969-7128},\n\tdoi = {10.1038/gt.2015.60},\n\tabstract = {Lentiviruses are the vectors of choice for many preclinical studies and clinical applications of gene therapy. Accurate measurement of biological vector titre  before treatment is a prerequisite for vector dosing, and the calculation of  vector integration sites per cell after treatment is as critical to the  characterisation of modified cell products as it is to long-term follow-up and  the assessment of risk and therapeutic efficiency in patients. These analyses are  typically based on quantitative real-time PCR (qPCR), but as yet compromise  accuracy and comparability between laboratories and experimental systems, the  former by using separate simplex reactions for the detection of endogene and  lentiviral sequences and the latter by designing different PCR assays for  analyses in human cells and animal disease models. In this study, we validate in  human and murine cells a qPCR system for the single-tube assessment of lentiviral  vector copy numbers that is suitable for analyses in at least 33 different  mammalian species, including human and other primates, mouse, pig, cat and  domestic ruminants. The established assay combines the accuracy of single-tube  quantitation by duplex qPCR with the convenience of one-off assay optimisation  for cross-species analyses and with the direct comparability of lentiviral  transduction efficiencies in different species.},\n\tlanguage = {eng},\n\tnumber = {1},\n\tjournal = {Gene therapy},\n\tauthor = {Christodoulou, I. and Patsali, P. and Stephanou, C. and Antoniou, M. and Kleanthous, M. and Lederer, C. W.},\n\tmonth = jan,\n\tyear = {2016},\n\tpmid = {26202078},\n\tpmcid = {PMC4705430},\n\tnote = {Place: England},\n\tkeywords = {*Genetic Vectors, *Real-Time Polymerase Chain Reaction, Animals, Base Sequence, Cell Line, Genetic Therapy, Humans, Lentivirus/*genetics, Mammals/genetics, Mice, Molecular Sequence Data, Sequence Alignment, Transduction, Genetic},\n\tpages = {113--118},\n}\n\n

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\n \n\n \n \n \n \n \n \n Beta testing: preclinical genome editing in β-globin disorders.\n \n \n \n \n\n\n \n Lederer, C. W., & Kleanthous, M.\n\n\n \n\n\n\n Cell and Gene Therapy Insights. December 2015.\n \n\n\n\n
\n\n\n\n \n \n $\"Beta$ Paper\n \n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{lederer_beta_2015,\n\ttitle = {Beta testing: preclinical genome editing in β-globin disorders},\n\tshorttitle = {Beta testing},\n\turl = {https://www.insights.bio/cell-and-gene-therapy-insights/journal/article/533/Beta-testing-preclinical-genome-editing-in-globin-disorders},\n\tdoi = {10.18609/cgti.2015.021},\n\tabstract = {The β-globin disorders β-thalassemia and sickle cell disease have been at the forefront of gene therapy development from its very inception. Owing to their frequency, severity and exceptionally well characterized molecular pathology, and to the availability of hematopoietic stem and progenitor cells as substrate for therapies, these disorders promise both fast insights into new methodologies and eventual return on investment. Accordingly, β-globinopathies are also a favorite subject of the nascent field of genome editing and its most pioneering approaches to achieving therapeutic functional correction of gene expression. Be it by mutation-specific correction, modulation of disease modifiers or site-specific gene addition, genome editing of β-globinopathies has already delivered significant insights into the design of synthetic nucleases and the suitability of different correction strategies. This Expert Insight reviews recent progress in the application of gene-editing tools and different model systems towards the establishment of new therapies for β-globin disorders.},\n\tlanguage = {en-US},\n\turldate = {2024-03-30},\n\tjournal = {Cell and Gene Therapy Insights},\n\tauthor = {Lederer, Carsten W. and Kleanthous, Marina},\n\tmonth = dec,\n\tyear = {2015},\n}\n\n

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\n \n\n \n \n \n \n \n Recent trends in the gene therapy of β-thalassemia.\n \n \n \n\n\n \n Finotti, A., Breda, L., Lederer, C. W., Bianchi, N., Zuccato, C., Kleanthous, M., Rivella, S., & Gambari, R.\n\n\n \n\n\n\n Journal of blood medicine, 6: 69–85. 2015.\n Place: New Zealand\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{finotti_recent_2015,\n\ttitle = {Recent trends in the gene therapy of β-thalassemia.},\n\tvolume = {6},\n\tissn = {1179-2736},\n\tdoi = {10.2147/JBM.S46256},\n\tabstract = {The β-thalassemias are a group of hereditary hematological diseases caused by over 300 mutations of the adult β-globin gene. Together with sickle cell anemia,  thalassemia syndromes are among the most impactful diseases in developing  countries, in which the lack of genetic counseling and prenatal diagnosis have  contributed to the maintenance of a very high frequency of these genetic diseases  in the population. Gene therapy for β-thalassemia has recently seen steadily  accelerating progress and has reached a crossroads in its development. Presently,  data from past and ongoing clinical trials guide the design of further clinical  and preclinical studies based on gene augmentation, while fundamental insights  into globin switching and new technology developments have inspired the  investigation of novel gene-therapy approaches. Moreover, human erythropoietic  stem cells from β-thalassemia patients have been the cellular targets of choice  to date whereas future gene-therapy studies might increasingly draw on induced  pluripotent stem cells. Herein, we summarize the most significant developments in  β-thalassemia gene therapy over the last decade, with a strong emphasis on the  most recent findings, for β-thalassemia model systems; for β-, γ-, and  anti-sickling β-globin gene addition and combinatorial approaches including the  latest results of clinical trials; and for novel approaches, such as  transgene-mediated activation of γ-globin and genome editing using designer  nucleases.},\n\tlanguage = {eng},\n\tjournal = {Journal of blood medicine},\n\tauthor = {Finotti, Alessia and Breda, Laura and Lederer, Carsten W. and Bianchi, Nicoletta and Zuccato, Cristina and Kleanthous, Marina and Rivella, Stefano and Gambari, Roberto},\n\tyear = {2015},\n\tpmid = {25737641},\n\tpmcid = {PMC4342371},\n\tnote = {Place: New Zealand},\n\tkeywords = {CRISPR, HbF induction, TALEN, Thalassemia, ZFN, gene therapy, genome editing, induced pluripotent stem cells, transcription factors},\n\tpages = {69--85},\n}\n\n

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\n \n\n \n \n \n \n \n Oxidative stress in β-thalassaemia and sickle cell disease.\n \n \n \n\n\n \n Voskou, S., Aslan, M., Fanis, P., Phylactides, M., & Kleanthous, M.\n\n\n \n\n\n\n Redox biology, 6: 226–239. December 2015.\n Place: Netherlands\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{voskou_oxidative_2015,\n\ttitle = {Oxidative stress in β-thalassaemia and sickle cell disease.},\n\tvolume = {6},\n\tcopyright = {Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.},\n\tissn = {2213-2317},\n\tdoi = {10.1016/j.redox.2015.07.018},\n\tabstract = {Sickle cell disease and β-thalassaemia are inherited haemoglobinopathies resulting in structural and quantitative changes in the β-globin chain. These  changes lead to instability of the generated haemoglobin or to globin chain  imbalance, which in turn impact the oxidative environment both intracellularly  and extracellularly. The ensuing oxidative stress and the inability of the body  to adequately overcome it are, to a large extent, responsible for the  pathophysiology of these diseases. This article provides an overview of the main  players and control mechanisms involved in the establishment of oxidative stress  in these haemoglobinopathies.},\n\tlanguage = {eng},\n\tjournal = {Redox biology},\n\tauthor = {Voskou, S. and Aslan, M. and Fanis, P. and Phylactides, M. and Kleanthous, M.},\n\tmonth = dec,\n\tyear = {2015},\n\tpmid = {26285072},\n\tpmcid = {PMC4543215},\n\tnote = {Place: Netherlands},\n\tkeywords = {Anemia, Sickle Cell/*blood/pathology, Anion Exchange Protein 1, Erythrocyte/chemistry/metabolism, Erythrocytes/metabolism/pathology, Heme/chemistry/metabolism, Hemeproteins/chemistry/metabolism, Humans, Iron Overload/*blood/pathology, Iron/metabolism, Oxidative Stress, Phosphatidylserines/chemistry/metabolism, Protein Stability, Reactive Oxygen Species/metabolism, Reperfusion Injury/*blood/pathology, Thrombophilia/*blood/pathology, beta-Globins/*metabolism, beta-Thalassemia/*blood/pathology},\n\tpages = {226--239},\n}\n\n

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\n \n\n \n \n \n \n \n The Molecular Basis of α-Thalassemia in the Qatari Pediatric Population.\n \n \n \n\n\n \n Kamal, M., Abu-Sirriya, S., Abu-Dayya, A., Al-Khatib, H., Abu-Ramadan, H., Petrou, M., Amer, A., Badii, R., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 39(5): 350–354. 2015.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kamal_molecular_2015,\n\ttitle = {The {Molecular} {Basis} of α-{Thalassemia} in the {Qatari} {Pediatric} {Population}.},\n\tvolume = {39},\n\tissn = {1532-432X 0363-0269},\n\tdoi = {10.3109/03630269.2015.1060606},\n\tabstract = {α-Thalassemia (α-thal) is widely reported in the Arabian Peninsula as one of the main causes of asymptomatic microcytic hypochromic red blood cells with or  without anemia in the pediatric population. This is the first study that provides  information about the molecular basis of α-thal in the Qatari population. Qatari  school children between the ages of 5 and 15, exhibiting laboratory findings  suggestive of microcytic anemia were pooled, and those with a mean corpuscular  volume (MCV) of {\\textless}80.0 fL and a hemoglobin (Hb) electropherogram that ruled out  β-thalassemia (β-thal), were narrowed down to a group of 127. This group was  screened for the -α(3.7) (rightward) deletion, and the α(-5 nt), α(polyA1)  (α(T-Saudi)), α(polyA2) mutations. A second group of randomly selected Qatari  individuals was also screened in order to determine the population's allele  frequency for the -α(3.7) deletion. Thirty-nine point four percent of the  individuals with microcytic hypochromic anemia were positive for the -α(3.7)  deletion (heterozygotes 30.0\\%, homozygotes 9.4\\%), 2.6\\% were positive for the  α(polyA1) deletion and 0.8\\% positive for the α(-5 nt) mutation. None of the  children exhibited changes in α(polyA2). Analysis of the random samples  determined that 26.4\\% were heterozygous and 4.5\\% homozygous for the -α(3.7)  deletion with a 17.7\\% allele frequency. Our results suggest that a significant  number of the Qatari pediatric population with microcytic hypochromic anemia are  carriers of α-thal mutations. However, 45.6\\% of the children failed to exhibit  any of the above four mutations tested. This suggests the possibility of other  mutations in the Qatari pediatric population that are yet to be elicited.},\n\tlanguage = {eng},\n\tnumber = {5},\n\tjournal = {Hemoglobin},\n\tauthor = {Kamal, Madeeha and Abu-Sirriya, Shaza and Abu-Dayya, Aseel and Al-Khatib, Hebah and Abu-Ramadan, Hadeel and Petrou, Miranda and Amer, Aliaa and Badii, Ramin and Kleanthous, Marina},\n\tyear = {2015},\n\tpmid = {26161810},\n\tnote = {Place: England},\n\tkeywords = {Erythrocyte Indices, Genotype, Humans, Mutation, Phenotype, Population Surveillance, Qatar, Qatar/epidemiology, alpha-Globins/*genetics, alpha-Thalassemia/diagnosis/*epidemiology/*genetics, pediatric population, α-Thalassemia (α-thal)},\n\tpages = {350--354},\n}\n\n

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\n \n\n \n \n \n \n \n Plerixafor+G-CSF-mobilized CD34+ cells represent an optimal graft source for thalassemia gene therapy.\n \n \n \n\n\n \n Karponi, G., Psatha, N., Lederer, C. W., Adair, J. E., Zervou, F., Zogas, N., Kleanthous, M., Tsatalas, C., Anagnostopoulos, A., Sadelain, M., Rivière, I., Stamatoyannopoulos, G., & Yannaki, E.\n\n\n \n\n\n\n Blood, 126(5): 616–619. July 2015.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{karponi_plerixaforg-csf-mobilized_2015,\n\ttitle = {Plerixafor+{G}-{CSF}-mobilized {CD34}+ cells represent an optimal graft source for thalassemia gene therapy.},\n\tvolume = {126},\n\tcopyright = {© 2015 by The American Society of Hematology.},\n\tissn = {1528-0020 0006-4971},\n\tdoi = {10.1182/blood-2015-03-629618},\n\tabstract = {Globin gene therapy requires abundant numbers of highly engraftable, autologous hematopoietic stem cells expressing curative levels of β-globin on  differentiation. In this study, CD34+ cells from 31 thalassemic patients  mobilized with hydroxyurea+granulocyte colony-stimulating factor (G-CSF), G-CSF,  Plerixafor, or Plerixafor+G-CSF were transduced with the TNS9.3.55 β-globin  lentivector and compared for transducibility and globin expression in vitro, as  well as engraftment potential in a xenogeneic model after partial myeloablation.  Transduction efficiency and vector copy number (VCN) averaged 48.4 ± 2.8\\% and  1.91 ± 0.04, respectively, whereas expression approximated the one-copy normal  β-globin output. Plerixafor+G-CSF cells produced the highest β-globin  expression/VCN. Long-term multilineage engraftment and persistent VCN and vector  expression was encountered in all xenografted groups, with  Plerixafor+G-CSF-mobilized cells achieving superior short-term engraftment rates,  with similar numbers of CD34+ cells transplanted. Overall, Plerixafor+G-CSF not  only allows high CD34+ cell yields but also provides increased β-globin  expression/VCN and enhanced early human chimerism under nonmyeloablative  conditions, thus representing an optimal graft for thalassemia gene therapy.},\n\tlanguage = {eng},\n\tnumber = {5},\n\tjournal = {Blood},\n\tauthor = {Karponi, Garyfalia and Psatha, Nikoletta and Lederer, Carsten Werner and Adair, Jennifer Eileen and Zervou, Fani and Zogas, Nikolaos and Kleanthous, Marina and Tsatalas, Constantinos and Anagnostopoulos, Achilles and Sadelain, Michel and Rivière, Isabelle and Stamatoyannopoulos, George and Yannaki, Evangelia},\n\tmonth = jul,\n\tyear = {2015},\n\tpmid = {26089395},\n\tpmcid = {PMC4520876},\n\tnote = {Place: United States},\n\tkeywords = {*Hematopoietic Stem Cell Transplantation, Animals, Antigens, CD34/metabolism, Benzylamines, Cyclams, Gene Dosage, Gene Expression, Genetic Therapy/*methods, Genetic Vectors, Granulocyte Colony-Stimulating Factor/*administration \\& dosage, Hematopoietic Stem Cell Mobilization/*methods, Hematopoietic Stem Cells/metabolism, Heterocyclic Compounds/administration \\& dosage, Heterografts, Humans, Mice, Mice, Knockout, Transplantation, Autologous, beta-Globins/genetics, beta-Thalassemia/genetics/*therapy},\n\tpages = {616--619},\n}\n\n

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\n \n\n \n \n \n \n \n The nucleotide-binding proteins Nubp1 and Nubp2 are negative regulators of ciliogenesis.\n \n \n \n\n\n \n Kypri, E., Christodoulou, A., Maimaris, G., Lethan, M., Markaki, M., Lysandrou, C., Lederer, C. W., Tavernarakis, N., Geimer, S., Pedersen, L. B., & Santama, N.\n\n\n \n\n\n\n Cellular and molecular life sciences: CMLS, 71(3): 517–538. February 2014.\n \n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kypri_nucleotide-binding_2014,\n\ttitle = {The nucleotide-binding proteins {Nubp1} and {Nubp2} are negative regulators of ciliogenesis},\n\tvolume = {71},\n\tissn = {1420-9071},\n\tdoi = {10.1007/s00018-013-1401-6},\n\tabstract = {Nucleotide-binding proteins Nubp1 and Nubp2 are MRP/MinD-type P-loop NTPases with sequence similarity to bacterial division site-determining proteins and are conserved, essential proteins throughout the Eukaryotes. They have been implicated, together with their interacting minus-end directed motor protein KIFC5A, in the regulation of centriole duplication in mammalian cells. Here we show that Nubp1 and Nubp2 are integral components of centrioles throughout the cell cycle, recruited independently of KIFC5A. We further demonstrate their localization at the basal body of the primary cilium in quiescent vertebrate cells or invertebrate sensory cilia, as well as in the motile cilia of mouse cells and in the flagella of Chlamydomonas. RNAi-mediated silencing of nubp-1 in C. elegans causes the formation of morphologically aberrant and additional cilia in sensory neurons. Correspondingly, downregulation of Nubp1 or Nubp2 in mouse quiescent NIH 3T3 cells markedly increases the number of ciliated cells, while knockdown of KIFC5A dramatically reduces ciliogenesis. Simultaneous double silencing of Nubp1 + KIFC5A restores the percentage of ciliated cells to control levels. We document the normal ciliary recruitment, during these silencing regimes, of basal body proteins critical for ciliogenesis, namely CP110, CEP290, cenexin, Chibby, AurA, Rab8, and BBS7. Interestingly, we uncover novel interactions of Nubp1 with several members of the CCT/TRiC molecular chaperone complex, which we find enriched at the basal body and recruited independently of the Nubps or KIFC5A. Our combined results for Nubp1, Nubp2, and KIFC5A and their striking effects on cilium formation suggest a central regulatory role for these proteins, likely involving CCT/TRiC chaperone activity, in ciliogenesis.},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {Cellular and molecular life sciences: CMLS},\n\tauthor = {Kypri, Elena and Christodoulou, Andri and Maimaris, Giannis and Lethan, Mette and Markaki, Maria and Lysandrou, Costas and Lederer, Carsten W. and Tavernarakis, Nektarios and Geimer, Stefan and Pedersen, Lotte B. and Santama, Niovi},\n\tmonth = feb,\n\tyear = {2014},\n\tpmid = {23807208},\n\tkeywords = {Animals, Blotting, Western, Cell Cycle, Centrioles, Chlamydomonas, Chromatography, Liquid, Cilia, Fluorescent Antibody Technique, GTP-Binding Proteins, Gene Knockdown Techniques, Immunohistochemistry, Immunoprecipitation, Intracellular Signaling Peptides and Proteins, Mice, Microscopy, Electron, Transmission, Microtubule-Associated Proteins, Molecular Chaperones, NIH 3T3 Cells, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Tandem Mass Spectrometry},\n\tpages = {517--538},\n}\n\n

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\n \n\n \n \n \n \n \n Genotyping of BCL11A and HBS1L-MYB SNPs associated with fetal haemoglobin levels: a SNaPshot minisequencing approach.\n \n \n \n\n\n \n Fanis, P., Kousiappa, I., Phylactides, M., & Kleanthous, M.\n\n\n \n\n\n\n BMC genomics, 15: 108. February 2014.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{fanis_genotyping_2014,\n\ttitle = {Genotyping of {BCL11A} and {HBS1L}-{MYB} {SNPs} associated with fetal haemoglobin levels: a {SNaPshot} minisequencing approach.},\n\tvolume = {15},\n\tissn = {1471-2164},\n\tdoi = {10.1186/1471-2164-15-108},\n\tabstract = {BACKGROUND: B-thalassaemia and sickle cell disease (SCD) are two of the most common monogenic diseases that are found in many populations worldwide. In both  disorders the clinical severity is highly variable, with the persistence of fetal  haemoglobin (HbF) being one of the major ameliorating factors. HbF levels are  affected by, amongst other factors, single nucleotide polymorphisms (SNPs) at the  BCL11A gene and the HBS1L-MYB intergenic region, which are located outside the  β-globin locus. For this reason, we developed two multiplex assays that allow the  genotyping of SNPs at these two genomic regions which have been shown to be  associated with variable HbF levels in different populations. RESULTS: Two  multiplex assays based on the SNaPshot minisequencing approach were developed.  The two assays can be used to simultaneous genotype twelve SNPs at the BCL11A  gene and sixteen SNPs at HBS1L-MYB intergenic region which were shown to modify  HbF levels. The different genotypes can be determined based on the position and  the fluorescent colour of the peaks in a single electropherogram. DNA sequencing  and restriction fragment length polymorphism (PCR-RFLP) assays were used to  verify genotyping results obtained by SNaPshot minisequencing. CONCLUSIONS: In  summary, we propose two multiplex assays based on the SNaPshot minisequencing  approach for the simultaneous identification of SNPs located at the BCL11A gene  and HBS1L-MYB intergenic region which have an effect on HbF levels. The assays  can be easily applied for accurate, time and cost efficient genotyping of the  selected SNPs in various populations.},\n\tlanguage = {eng},\n\tjournal = {BMC genomics},\n\tauthor = {Fanis, Pavlos and Kousiappa, Ioanna and Phylactides, Marios and Kleanthous, Marina},\n\tmonth = feb,\n\tyear = {2014},\n\tpmid = {24502199},\n\tpmcid = {PMC3922441},\n\tnote = {Place: England},\n\tkeywords = {*Polymorphism, Single Nucleotide, Carrier Proteins/*genetics/metabolism, DNA, Intergenic/*genetics/metabolism, Fetal Hemoglobin/analysis/*genetics, GTP-Binding Proteins/genetics, Genotype, HSP70 Heat-Shock Proteins/genetics, Humans, Multiplex Polymerase Chain Reaction, Nuclear Proteins/*genetics/metabolism, Peptide Elongation Factors/genetics, Proto-Oncogene Proteins c-myb/genetics, Repressor Proteins, Sequence Analysis, DNA/*methods},\n\tpages = {108},\n}\n\n

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\n \n\n \n \n \n \n \n The first family with Tay-Sachs disease in Cyprus: Genetic analysis reveals a nonsense (c.78G\\textgreaterA) and a silent (c.1305C\\textgreaterT) mutation and allows preimplantation genetic diagnosis.\n \n \n \n\n\n \n Georgiou, T., Christopoulos, G., Anastasiadou, V., Hadjiloizou, S., Cregeen, D., Jackson, M., Mavrikiou, G., Kleanthous, M., & Drousiotou, A.\n\n\n \n\n\n\n Meta gene, 2: 200–205. December 2014.\n Place: Netherlands\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{georgiou_first_2014,\n\ttitle = {The first family with {Tay}-{Sachs} disease in {Cyprus}: {Genetic} analysis reveals a nonsense (c.{78G}{\\textgreater}{A}) and a silent (c.{1305C}{\\textgreater}{T}) mutation and allows preimplantation  genetic diagnosis.},\n\tvolume = {2},\n\tissn = {2214-5400},\n\tdoi = {10.1016/j.mgene.2014.01.007},\n\tabstract = {Tay-Sachs disease (TSD) is a recessively inherited neurodegenerative disorder caused by mutations in the HEXA gene resulting in β-hexosaminidase A (HEX A)  deficiency and neuronal accumulation of GM2 ganglioside. We describe the first  patient with Tay-Sachs disease in the Cypriot population, a juvenile case which  presented with developmental regression at the age of five. The diagnosis was  confirmed by measurement of HEXA activity in plasma, peripheral leucocytes and  fibroblasts. Sequencing the HEXA gene resulted in the identification of two  previously described mutations: the nonsense mutation c.78G{\\textgreater}A (p.Trp26X) and the  silent mutation c.1305C{\\textgreater}T (p.=). The silent mutation was reported once before in  a juvenile TSD patient of West Indian origin with an unusually mild phenotype.  The presence of this mutation in another juvenile TSD patient provides further  evidence that it is a disease-causing mutation. Successful preimplantation  genetic diagnosis (PGD) and prenatal follow-up were provided to the couple.},\n\tlanguage = {eng},\n\tjournal = {Meta gene},\n\tauthor = {Georgiou, Theodoros and Christopoulos, George and Anastasiadou, Violetta and Hadjiloizou, Stavros and Cregeen, David and Jackson, Marie and Mavrikiou, Gavriella and Kleanthous, Marina and Drousiotou, Anthi},\n\tmonth = dec,\n\tyear = {2014},\n\tpmid = {25606403},\n\tpmcid = {PMC4287815},\n\tnote = {Place: Netherlands},\n\tkeywords = {ADO, Allele Dropout, CVS, Chorionic Villus Sampling, Cyprus, EEG, Electroencephalogram, GM2 gangliosidosis, Juvenile Tay-Sachs disease, MLPA, Multiplex Ligation Dependent Probe Amplification, MRI, Magnetic Resonance Imaging, PCR, Polymerase Chain Reaction, PGD, Preimplantation Genetic Diagnosis, Preimplantation genetic diagnosis, STR, Short Tandem Repeat, Silent mutation, TSD, Tay-Sachs disease, β-Hexosaminidase A},\n\tpages = {200--205},\n}\n\n

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\n \n\n \n \n \n \n \n Hb Famagusta–analysis of a novel δ-globin chain variant [HBD:c.60C\\textgreaterA] in four families with diverse globin genotypes.\n \n \n \n\n\n \n Lederer, C. W., Pavlou, E., Makariou, C., Hadjilambi, G., Andreou, N., Hadjigavriel, M., Kolnagou, A., Sitarou, M., Christou, S., & Kleanthous, M.\n\n\n \n\n\n\n Annals of hematology, 93(9): 1625–1627. September 2014.\n Place: Germany\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{lederer_hb_2014,\n\ttitle = {Hb {Famagusta}--analysis of a novel δ-globin chain variant [{HBD}:c.{60C}{\\textgreater}{A}] in four families with diverse globin genotypes.},\n\tvolume = {93},\n\tissn = {1432-0584 0939-5555},\n\tdoi = {10.1007/s00277-013-1996-6},\n\tlanguage = {eng},\n\tnumber = {9},\n\tjournal = {Annals of hematology},\n\tauthor = {Lederer, Carsten W. and Pavlou, Eleni and Makariou, Christiana and Hadjilambi, Georgia and Andreou, Nicoletta and Hadjigavriel, Michael and Kolnagou, Annita and Sitarou, Maria and Christou, Soteroulla and Kleanthous, Marina},\n\tmonth = sep,\n\tyear = {2014},\n\tpmid = {24452365},\n\tnote = {Place: Germany},\n\tkeywords = {*Polymorphism, Single Nucleotide, Base Sequence, Family, Female, Genetic Variation, Genotype, Greece, Hemoglobins, Abnormal/chemistry/*genetics, Humans, Male, Models, Molecular, delta-Globins/chemistry/*genetics},\n\tpages = {1625--1627},\n}\n\n

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\n \n 2013\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n Liver iron and serum ferritin levels are misleading for estimating cardiac, pancreatic, splenic and total body iron load in thalassemia patients: factors influencing the heterogenic distribution of excess storage iron in organs as identified by MRI T2*.\n \n \n \n\n\n \n Kolnagou, A., Natsiopoulos, K., Kleanthous, M., Ioannou, A., & Kontoghiorghes, G. J.\n\n\n \n\n\n\n Toxicology mechanisms and methods, 23(1): 48–56. January 2013.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kolnagou_liver_2013,\n\ttitle = {Liver iron and serum ferritin levels are misleading for estimating cardiac, pancreatic, splenic and total body iron load in thalassemia patients: factors  influencing the heterogenic distribution of excess storage iron in organs as  identified by {MRI} {T2}*.},\n\tvolume = {23},\n\tissn = {1537-6524 1537-6516},\n\tdoi = {10.3109/15376516.2012.727198},\n\tabstract = {A comparative assessment of excess storage iron distribution in the liver, heart, spleen and pancreas of β-thalassemia major (β-ΤΜ) patients has been carried out  using magnetic resonance imaging (MRI) relaxation times T2*. The β-ΤΜ patients  (8-40 years, 11 males, 9 females) had variable serum ferritin levels (394-5603  μg/L) and were treated with deferoxamine (n = 10), deferiprone (n = 5) and  deferoxamine/deferiprone combination (n = 5). MRI T2* assessment revealed that  excess iron is not proportionally distributed among the organs but is stored at  different concentrations in each organ and the distribution is different for each  β-ΤΜ patient. There is random variation in the distribution of excess storage  iron from normal to severe levels in each organ among the β-ΤΜ patients by  comparison to the same organs of ten normal volunteers. The correlation of serum  ferritin with T2* was for spleen (r = -0.81), liver (r = -0.63), pancreas (r =  -0.33) and none with heart. Similar trend was observed in the correlation of  liver T2* with the T2* of spleen (r = 0.62), pancreas (r = 0.61) and none with  heart. These studies contradict previous assumptions that serum ferritin and  liver iron concentration is proportional to the total body iron stores in β-ΤΜ  and especially cardiac iron load. The random variation in the concentration of  iron in the organs of β-ΤΜ patients appears to be related to the chelation  protocol, organ function, genetic, dietary, pharmacological and other factors.  Monitoring of the iron load for all the organs is recommended for each β-ΤΜ  patient.},\n\tlanguage = {eng},\n\tnumber = {1},\n\tjournal = {Toxicology mechanisms and methods},\n\tauthor = {Kolnagou, Annita and Natsiopoulos, Konstantinos and Kleanthous, Marios and Ioannou, Alexia and Kontoghiorghes, George J.},\n\tmonth = jan,\n\tyear = {2013},\n\tpmid = {22943064},\n\tnote = {Place: England},\n\tkeywords = {Adolescent, Adult, Body Burden, Child, Deferiprone, Deferoxamine/therapeutic use, Drug Therapy, Combination, Female, Ferritins/*blood, Humans, Iron Chelating Agents/therapeutic use, Iron Overload/drug therapy/etiology/metabolism, Iron/analysis/*metabolism, Liver/chemistry/*metabolism, Magnetic Resonance Imaging, Male, Myocardium/chemistry/*metabolism, Pancrelipase/chemistry/*metabolism, Pyridones/therapeutic use, Spleen/chemistry/*metabolism, Young Adult, beta-Thalassemia/complications/drug therapy/*metabolism},\n\tpages = {48--56},\n}\n\n

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\n \n\n \n \n \n \n \n A minimal set of SNPs for the noninvasive prenatal diagnosis of β-thalassaemia.\n \n \n \n\n\n \n Papasavva, T. E., Lederer, C. W., Traeger-Synodinos, J., Mavrou, A., Kanavakis, E., Ioannou, C., Makariou, C., & Kleanthous, M.\n\n\n \n\n\n\n Annals of human genetics, 77(2): 115–124. March 2013.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{papasavva_minimal_2013,\n\ttitle = {A minimal set of {SNPs} for the noninvasive prenatal diagnosis of β-thalassaemia.},\n\tvolume = {77},\n\tcopyright = {© 2013 Blackwell Publishing Ltd/University College London.},\n\tissn = {1469-1809 0003-4800},\n\tdoi = {10.1111/ahg.12004},\n\tabstract = {β-thalassaemia is one of the commonest autosomal recessive single-gene disorders worldwide. Prenatal tests use invasive methods, posing a risk for the pregnancy  itself. Development of a noninvasive prenatal diagnostic method is, therefore, of  paramount importance. The aim of the present study is to identify  high-heterozygote informative single-nucleotide polymorphisms (SNPs), suitable  for the development of noninvasive prenatal diagnosis (NIPD) of β-thalassaemia.  SNP genotyping analysis was performed on 75 random samples from the Cypriot  population for 140 SNPs across the β-globin cluster. Shortlisted, highly  heterozygous SNPs were then examined in 101 carrier families for their  applicability in the noninvasive detection of paternally inherited alleles.  Forty-nine SNPs displayed more than 6\\% heterozygosity and were selected for NIPD  analysis, revealing 72.28\\% of the carrier families eligible for qualitative  SNP-based NIPD, and 92\\% for quantitative detection. Moreover, inference of  haplotypes showed predominant haplotypes and many subhaplotypes with sufficient  prevalence for diagnostic exploitation. SNP-based analyses are sensitive and  specific for the detection of the paternally inherited allele in maternal plasma.  This study provides proof of concept for this approach, highlighting its  superiority to NIPD based on single markers and thus providing a blueprint for  the general development of noninvasive prenatal diagnostic assays for  β-thalassaemia.},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {Annals of human genetics},\n\tauthor = {Papasavva, Thessalia E. and Lederer, Carsten W. and Traeger-Synodinos, Jan and Mavrou, Ariadne and Kanavakis, Emmanuel and Ioannou, Christiana and Makariou, Christiana and Kleanthous, Marina},\n\tmonth = mar,\n\tyear = {2013},\n\tpmid = {23362932},\n\tnote = {Place: England},\n\tkeywords = {*Polymorphism, Single Nucleotide, Female, Haplotypes, Heterozygote, Humans, Pregnancy, Prenatal Diagnosis/*methods, beta-Globins/*genetics, beta-Thalassemia/*diagnosis/genetics},\n\tpages = {115--124},\n}\n\n

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\n \n 2012\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n KLF10 gene expression is associated with high fetal hemoglobin levels and with response to hydroxyurea treatment in β-hemoglobinopathy patients.\n \n \n \n\n\n \n Borg, J., Phylactides, M., Bartsakoulia, M., Tafrali, C., Lederer, C., Felice, A. E., Papachatzopoulou, A., Kourakli, A., Stavrou, E. F., Christou, S., Hou, J., Karkabouna, S., Lappa-Manakou, C., Ozgur, Z., van Ijcken, W., von Lindern, M., Grosveld, F. G., Georgitsi, M., Kleanthous, M., Philipsen, S., & Patrinos, G. P.\n\n\n \n\n\n\n Pharmacogenomics, 13(13): 1487–1500. October 2012.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{borg_klf10_2012,\n\ttitle = {{KLF10} gene expression is associated with high fetal hemoglobin levels and with response to hydroxyurea treatment in β-hemoglobinopathy patients.},\n\tvolume = {13},\n\tissn = {1744-8042 1462-2416},\n\tdoi = {10.2217/pgs.12.125},\n\tabstract = {AIM: In humans, fetal hemoglobin (HbF) production is controlled by many intricate mechanisms that, to date, remain only partly understood. PATIENTS \\& METHODS:  Pharmacogenomic analysis of the effects of hydroxyurea (HU) on HbF production was  undertaken in a collection of Hellenic β-thalassemia and sickle cell disease  (SCD) compound heterozygotes and a collection of healthy and  KLF1-haploinsufficient Maltese adults, to identify genomic signatures that follow  high HbF patterns. RESULTS: KLF10 emerged as a top candidate. Moreover, genotype  analysis of β-thalassemia major and intermedia patients and an independent cohort  of β-thalassemia/SCD compound heterozygous patients that do or do not respond to  HU treatment showed that the homozygous mutant state of a tagSNP in the KLF10  3'UTR is not present in β-thalassemia intermedia patients and is underrepresented  in β-thalassemia/SCD compound heterozygous patients that respond well to HU  treatment. CONCLUSION: These data suggest that KLF10 may constitute a  pharmacogenomic marker to discriminate between response and nonresponse to HU  treatment.},\n\tlanguage = {eng},\n\tnumber = {13},\n\tjournal = {Pharmacogenomics},\n\tauthor = {Borg, Joseph and Phylactides, Marios and Bartsakoulia, Marina and Tafrali, Christina and Lederer, Carsten and Felice, Alexander E. and Papachatzopoulou, Adamantia and Kourakli, Alexandra and Stavrou, Eleana F. and Christou, Soteroula and Hou, Jun and Karkabouna, Sophia and Lappa-Manakou, Christina and Ozgur, Zeliha and van Ijcken, Wilfred and von Lindern, Marieke and Grosveld, Frank G. and Georgitsi, Marianthi and Kleanthous, Marina and Philipsen, Sjaak and Patrinos, George P.},\n\tmonth = oct,\n\tyear = {2012},\n\tpmid = {23057549},\n\tnote = {Place: England},\n\tkeywords = {3' Untranslated Regions, Adult, Anemia, Sickle Cell/blood/drug therapy/genetics, Antisickling Agents/therapeutic use, Early Growth Response Transcription Factors/*genetics, Erythroid Precursor Cells/metabolism, Female, Fetal Hemoglobin/genetics/*metabolism, Gene Expression, Genetic Markers, Hemoglobinopathies/blood/*drug therapy/*genetics, Heterozygote, Humans, Hydroxyurea/*therapeutic use, Kruppel-Like Transcription Factors/*genetics, Male, Polymorphism, Single Nucleotide, Retrospective Studies, Transcriptome, beta-Thalassemia/blood/drug therapy/genetics},\n\tpages = {1487--1500},\n}\n\n

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\n \n\n \n \n \n \n \n Microsatellite markers within the α-globin gene cluster for robust preimplantation genetic diagnosis of severe α-thalassemia syndromes in Mediterranean populations.\n \n \n \n\n\n \n Destouni, A., Christopoulos, G., Vrettou, C., Kakourou, G., Kleanthous, M., Traeger-Synodinos, J., & Kanavakis, E.\n\n\n \n\n\n\n Hemoglobin, 36(3): 253–264. 2012.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{destouni_microsatellite_2012,\n\ttitle = {Microsatellite markers within the α-globin gene cluster for robust preimplantation genetic diagnosis of severe α-thalassemia syndromes in  {Mediterranean} populations.},\n\tvolume = {36},\n\tissn = {1532-432X 0363-0269},\n\tdoi = {10.3109/03630269.2012.666512},\n\tabstract = {In this study we report the development of a generic protocol for preimplantation genetic diagnosis (PGD) of severe α-thalassemia (α-thal) syndromes in α-thal  carrier couples of Mediterranean origin. The in silico identification and design  of primers for multiplex analysis of short tandem repeats (STRs), was followed by  the optimization of polymerase chain reaction (PCR) conditions for multiplexed  STR analysis within the α-globin gene cluster (16p3.3) and subsequent  optimization and validation of a single-cell multiplex reaction including the  selected STRs. Three simple dinucleotide repeats were selected based on their  rate of heterozygosity, multiplex PCR efficiency and product size, and location  within the α-globin gene cluster. The multiplex PCR was optimized in single  lymphocytes with PCR efficiency ranging from 92.5 to 98\\% and an allele drop-out  (ADO) rate of 0 to 9.0\\% for the three loci. The optimized method was applied in  two clinical PGD cycles and genotypes were achieved in 17 out of 18 blastomeres  (94\\%). Transfer of unaffected embryos led to a singleton pregnancy in one of the  two couples. The triplex PCR validated for Greek and Cypriot populations is a  robust generic method for α-thal PGD.},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {Hemoglobin},\n\tauthor = {Destouni, Aspasia and Christopoulos, George and Vrettou, Christina and Kakourou, Georgia and Kleanthous, Marina and Traeger-Synodinos, Jan and Kanavakis, Emmanuel},\n\tyear = {2012},\n\tpmid = {22452522},\n\tnote = {Place: England},\n\tkeywords = {*Multigene Family, Cyprus, Female, Greece, Humans, Male, Microsatellite Repeats/*genetics, Multiplex Polymerase Chain Reaction/methods, Pregnancy, Pregnancy Complications, Hematologic/diagnosis/genetics, Preimplantation Diagnosis/*methods, Reproducibility of Results, Sensitivity and Specificity, Syndrome, alpha-Globins/*genetics, alpha-Thalassemia/*diagnosis/*genetics},\n\tpages = {253--264},\n}\n\n

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\n \n 2011\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n A novel β-globin gene CAP site mutation in association with the 92+1G>A mutation was found in a thalassemia intermedia patient.\n \n \n \n\n\n \n Letitia, D., Rodica, T., Xenia, F., Daniel, C., Florentina, V., Marina, K., Ciprian, T., & Davila, C.\n\n\n \n\n\n\n . January 2011.\n MAG ID: 2185109674\n\n\n\n
\n\n\n\n \n\n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n\n\n\n

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@article{letitia_novel_2011,\n\ttitle = {A novel β-globin gene {CAP} site mutation in association with the 92+{1G}\\&\\#62;{A} mutation was found in a thalassemia intermedia patient},\n\tabstract = {This paper describes a novel β-thalassemia mutation 3 base pairs downstream of the CAP site of the β- globin gene, -48A{\\textgreater}T. The proband, an 11-year-old Romanian girl, is a compound heterozygote for this mutation and the common 92+1G{\\textgreater}A β(0) thalassemia mutation. She has a mild thalassemia intermedia phenotype and is transfusion independent. Her mother (N/92+1G{\\textgreater}A) has total hemoglobin levels of 11.1 g/dl, while the proband's father (N/-48A{\\textgreater}T) has normal hematological indices. These data indicate that this novel CAP site mutation may play a role in the phenotypic expression of the disease in this case.},\n\tauthor = {Letitia, Dan and Rodica, Talmaci and Xenia, Feleki and Daniel, Coriu and Florentina, Vladareanu and Marina, Kleanthous and Ciprian, Tecuceanu and Davila, Carol},\n\tmonth = jan,\n\tyear = {2011},\n\tnote = {MAG ID: 2185109674},\n}\n\n

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\n \n\n \n \n \n \n \n Efficacy, compliance and toxicity factors are affecting the rate of normalization of body iron stores in thalassemia patients using the deferiprone and deferoxamine combination therapy.\n \n \n \n\n\n \n Kolnagou, A., Kleanthous, M., & Kontoghiorghes, G. J.\n\n\n \n\n\n\n Hemoglobin, 35(3): 186–198. 2011.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kolnagou_efficacy_2011,\n\ttitle = {Efficacy, compliance and toxicity factors are affecting the rate of normalization of body iron stores in thalassemia patients using the deferiprone and  deferoxamine combination therapy.},\n\tvolume = {35},\n\tissn = {1532-432X 0363-0269},\n\tdoi = {10.3109/03630269.2011.576153},\n\tabstract = {The international committee on chelation (ICOC) of deferiprone (L1) and deferoxamine (DFO) combination therapy was the first protocol reported to have  achieved normal range body iron store levels (NRBISL) in β-thalassemia major  (β-TM) patients. A follow-up study in eight β-TM patients has been designed to  investigate the factors affecting the rate of iron removal leading to NRBISL. The  patients had variable serum ferritin [mean ± SE (standard error) =1692 ± 366,  range 539-3845 μg/L)] and magnetic resonance imaging (MRI) T2* relaxation times  cardiac (mean ± SE =11.1 ± 2.5, range 4.5-24.2 ms) and liver (mean ± SE = 4.3 ±  1.8, range 1.4-14 ms). Organ function, blood and other biochemical parameters  were regularly monitored for toxicity. The ICOC L1 (80-100 mg/kg/day) and DFO  (40-60 mg/kg, at least 3 days per week) combination therapy caused an increase in  cardiac (mean ± SE =30.2 ± 2.3, range 22-41 ms) and liver (mean ± SE =27.6 ± 2.8,  range 9.1-35 ms) T2* and reduction in serum ferritin (mean ± SE = 158 ± 49, range  40-421 μg/L) to within the NRBISL. The rate of normalization was variable and in  one case was achieved within 9 months, whereas the longest was about 3 years. The  initial iron load, the rate of transfusions, the combination dose protocol and  the level of compliance were the major factors affecting the rate of  normalization of the iron stores. No serious toxicity was observed during the  study period, which lasted a total of 24.7 patient years.},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {Hemoglobin},\n\tauthor = {Kolnagou, Annita and Kleanthous, Marios and Kontoghiorghes, George J.},\n\tyear = {2011},\n\tpmid = {21599431},\n\tnote = {Place: England},\n\tkeywords = {Deferiprone, Deferoxamine/*therapeutic use, Drug Therapy, Combination, Ferritins/blood/drug effects, Humans, Iron Chelating Agents/pharmacokinetics/therapeutic use/toxicity, Iron/*metabolism, Liver/metabolism, Longitudinal Studies, Magnetic Resonance Imaging, Myocardium/metabolism, Pyridones/*therapeutic use, beta-Thalassemia/*drug therapy/metabolism},\n\tpages = {186--198},\n}\n\n

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\n \n 2010\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n Reduction of body iron stores to normal range levels in thalassaemia by using a deferiprone/deferoxamine combination and their maintenance thereafter by deferiprone monotherapy.\n \n \n \n\n\n \n Kolnagou, A., Kleanthous, M., & Kontoghiorghes, G. J.\n\n\n \n\n\n\n European journal of haematology, 85(5): 430–438. November 2010.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kolnagou_reduction_2010,\n\ttitle = {Reduction of body iron stores to normal range levels in thalassaemia by using a deferiprone/deferoxamine combination and their maintenance thereafter by  deferiprone monotherapy.},\n\tvolume = {85},\n\tcopyright = {© 2010 John Wiley \\& Sons A/S.},\n\tissn = {1600-0609 0902-4441},\n\tdoi = {10.1111/j.1600-0609.2010.01499.x},\n\tabstract = {BACKGROUND: Iron overload and toxicity is the major cause of morbidity and mortality in thalassaemia patients. New chelating drug protocols are necessary to  treat completely transfusional iron overload and eliminate associated toxicity.  Appropriate deferiprone/deferoxamine combinations could achieve this goal.  METHODS: A single-centre, single-armed, proof-of-concept study of the combination  of deferiprone (75-100 mg/kg/d) and deferoxamine (40-60 mg/kg, at least 3 d per  week) was carried out in eight patients with thalassaemia major (four men and  four women) for 21-68 months. The patients were previously treated with  deferoxamine and had variable serum ferritin [geometric (G) mean ± SD = 1446 ±  1035 μg/L] and magnetic resonance imaging relaxation times T2* cardiac (Gmean ±  SD = 10.32 ± 6.72 ms) and liver (G mean ± SD = 3.77 ± 4.69 ms). The use of  deferiprone (80-100 mg/kg/d) continued for 7-26 months in seven of the eight  patients following the combination therapy. Organ function, blood and other  biochemical parameters were monitored for toxicity. RESULTS:   The  deferiprone/deferoxamine combination caused an absolute value increase in cardiac  (G mean ± SD = 29.6 ± 6.6 ms, P {\\textless} 0.00076) and liver (G mean ± SD = 25.9 ± 8.07  ms, P {\\textless} 0.00075) T2* and reduction in serum ferritin (G mean ± SD = 114.7 ± 139.8  μg/L, P {\\textless} 0.0052) to within the normal body iron store range levels. In two  cases, normalisation was achieved within a year. Deferiprone monotherapy was  sufficient thereafter in maintaining normal range cardiac (G mean ± SD = 31.4 ±  5.25 ms, P {\\textless} 0.79) and liver (G mean ± SD = 26.2 ± 12.4 ms, P {\\textless} 0.58) T2* and  normal serum ferritin (G mean ± SD = 150.7 ± 159.1, μg/L, P {\\textless} 0.17) in five of  the seven patients. No serious toxicity was observed. CONCLUSION:   Transfusional  iron overload in patients with thalassaemia could be reduced to normal body iron  range levels using effective deferiprone/deferoxamine combinations. These levels  could be maintained using deferiprone monotherapy.},\n\tlanguage = {eng},\n\tnumber = {5},\n\tjournal = {European journal of haematology},\n\tauthor = {Kolnagou, Annita and Kleanthous, Marios and Kontoghiorghes, George J.},\n\tmonth = nov,\n\tyear = {2010},\n\tpmid = {20662901},\n\tnote = {Place: England},\n\tkeywords = {Adult, Deferiprone, Deferoxamine/*administration \\& dosage, Drug Therapy, Combination, Female, Ferritins/blood, Humans, Iron Chelating Agents/therapeutic use, Iron Overload/*drug therapy, Iron/metabolism, Liver/metabolism, Magnetic Resonance Imaging, Male, Myocardium/metabolism, Pyridones/*administration \\& dosage, Siderophores/therapeutic use, Thalassemia/*drug therapy, Treatment Outcome},\n\tpages = {430--438},\n}\n\n

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\n BACKGROUND: Iron overload and toxicity is the major cause of morbidity and mortality in thalassaemia patients. New chelating drug protocols are necessary to treat completely transfusional iron overload and eliminate associated toxicity. Appropriate deferiprone/deferoxamine combinations could achieve this goal. METHODS: A single-centre, single-armed, proof-of-concept study of the combination of deferiprone (75-100 mg/kg/d) and deferoxamine (40-60 mg/kg, at least 3 d per week) was carried out in eight patients with thalassaemia major (four men and four women) for 21-68 months. The patients were previously treated with deferoxamine and had variable serum ferritin [geometric (G) mean ± SD = 1446 ± 1035 μg/L] and magnetic resonance imaging relaxation times T2* cardiac (Gmean ± SD = 10.32 ± 6.72 ms) and liver (G mean ± SD = 3.77 ± 4.69 ms). The use of deferiprone (80-100 mg/kg/d) continued for 7-26 months in seven of the eight patients following the combination therapy. Organ function, blood and other biochemical parameters were monitored for toxicity. RESULTS: The deferiprone/deferoxamine combination caused an absolute value increase in cardiac (G mean ± SD = 29.6 ± 6.6 ms, P \\textless 0.00076) and liver (G mean ± SD = 25.9 ± 8.07 ms, P \\textless 0.00075) T2* and reduction in serum ferritin (G mean ± SD = 114.7 ± 139.8 μg/L, P \\textless 0.0052) to within the normal body iron store range levels. In two cases, normalisation was achieved within a year. Deferiprone monotherapy was sufficient thereafter in maintaining normal range cardiac (G mean ± SD = 31.4 ± 5.25 ms, P \\textless 0.79) and liver (G mean ± SD = 26.2 ± 12.4 ms, P \\textless 0.58) T2* and normal serum ferritin (G mean ± SD = 150.7 ± 159.1, μg/L, P \\textless 0.17) in five of the seven patients. No serious toxicity was observed. CONCLUSION: Transfusional iron overload in patients with thalassaemia could be reduced to normal body iron range levels using effective deferiprone/deferoxamine combinations. These levels could be maintained using deferiprone monotherapy.\n

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\n \n\n \n \n \n \n \n Compounds of the anthracycline family of antibiotics elevate human gamma-globin expression both in erythroid cultures and in a transgenic mouse model.\n \n \n \n\n\n \n Spyrou, P., Phylactides, M., Lederer, C. W., Kithreotis, L., Kirri, A., Christou, S., Kkolou, E., Kanavakis, E., Anagnou, N. P., Stamatoyannopoulos, G., & Kleanthous, M.\n\n\n \n\n\n\n Blood cells, molecules & diseases, 44(2): 100–106. April 2010.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{spyrou_compounds_2010,\n\ttitle = {Compounds of the anthracycline family of antibiotics elevate human gamma-globin expression both in erythroid cultures and in a transgenic mouse model.},\n\tvolume = {44},\n\tcopyright = {Copyright (c) 2009 Elsevier Inc. All rights reserved.},\n\tissn = {1096-0961 1079-9796},\n\tdoi = {10.1016/j.bcmd.2009.10.008},\n\tabstract = {We examined the effect of the anthracyclines aclarubicin, bleomycin, daunorubicin, doxorubicin and idarubicin on human gamma- and beta-globin promoter  activity in an in vitro luciferase assay, ex vivo in erythroid cultures and in  vivo in transgenic mice carrying the human gamma-globin gene. Effects in  erythroid liquid cultures derived from healthy donors were assayed by evaluating  HbF production with high performance liquid chromatography and by measuring mRNA  levels of the globin genes and the proportion of erythroblasts containing HbF.  Compounds testing positive in the in vitro and ex vivo assays were applied to  erythroid cultures derived from thalassaemic patients. Doxorubicin, idarubicin  and daunorubicin increased HbF production in cultures of both, healthy and  thalassaemic donors. Daunorubicin induced HbF in thalassaemic cells ex vivo with  the highest statistical significance and, importantly and in contrast to the  clinical HbF inducer hydroxyurea, showed specific induction of gamma-globin  without associated induction of alpha-globin. Daunorubicin was screened in  transgenic mice carrying the human (A)gamma-globin gene, and it resulted in  increased (A)gamma-globin mRNA levels. Our results indicate that anthracyclines  are a promising group of compounds with the potential to provide lead substances  for the synthesis of new agents with clinical applications as gamma-globin gene  inducers. In parallel, future studies of the epigenetic effects of the five  anthracyclines on the beta-globin locus will generate possible mechanistic leads  on the regulation of the globin genes.},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {Blood cells, molecules \\& diseases},\n\tauthor = {Spyrou, Pandelis and Phylactides, Marios and Lederer, Carsten W. and Kithreotis, Lucas and Kirri, Andriani and Christou, Soteroulla and Kkolou, Elena and Kanavakis, Emanuel and Anagnou, Nicholas P. and Stamatoyannopoulos, George and Kleanthous, Marina},\n\tmonth = apr,\n\tyear = {2010},\n\tpmid = {19914848},\n\tnote = {Place: United States},\n\tkeywords = {Animals, Anthracyclines/administration \\& dosage/*pharmacology, Anti-Bacterial Agents/administration \\& dosage/*pharmacology, Cell Line, Cells, Cultured, Erythroid Cells/drug effects, Fetal Hemoglobin/genetics/metabolism, Gene Expression Regulation/*drug effects, Humans, Mice, Mice, Transgenic, Promoter Regions, Genetic/drug effects, RNA, Messenger/genetics, Thalassemia/drug therapy/genetics, beta-Globins/genetics, gamma-Globins/*genetics},\n\tpages = {100--106},\n}\n\n

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\n \n 2009\n \n \n (3)\n \n \n

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\n \n\n \n \n \n \n \n Prenatal diagnosis of hemoglobin disorders: present and future strategies.\n \n \n \n\n\n \n Harteveld, C. L., Kleanthous, M., & Traeger-Synodinos, J.\n\n\n \n\n\n\n Clinical biochemistry, 42(18): 1767–1779. December 2009.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{harteveld_prenatal_2009,\n\ttitle = {Prenatal diagnosis of hemoglobin disorders: present and future strategies.},\n\tvolume = {42},\n\tissn = {1873-2933 0009-9120},\n\tdoi = {10.1016/j.clinbiochem.2009.07.001},\n\tabstract = {Prenatal diagnosis for the hemoglobinopathies based on molecular analysis of trophoblast or amniocyte DNA has accumulated around 30 years of experience,  following the first applications in the 1970s. As the first monogenic diseases to  be characterized at the molecular level the disorders of hemoglobin synthesis  (thalassemias and hemoglobinopathies) have been used as a prototype for the  development of many techniques of mutation detection, and consequently there are  numerous PCR-based techniques described in the literature that can be used for  prenatal diagnosis of the globin gene mutations. This review describes the most  commonly used current methods, as well as the most promising newer methods. In  addition, it outlines the newer application of preimplantation genetic diagnosis  (PGD) and the state of progress in the developing field of non-invasive prenatal  diagnosis.},\n\tlanguage = {eng},\n\tnumber = {18},\n\tjournal = {Clinical biochemistry},\n\tauthor = {Harteveld, Cornelis L. and Kleanthous, Marina and Traeger-Synodinos, Joanne},\n\tmonth = dec,\n\tyear = {2009},\n\tpmid = {19607819},\n\tnote = {Place: United States},\n\tkeywords = {DNA/genetics, Female, Hemoglobinopathies/*diagnosis, Humans, Mutation/genetics, Pregnancy, Preimplantation Diagnosis/trends, Prenatal Diagnosis/*methods/*trends},\n\tpages = {1767--1779},\n}\n\n

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\n \n\n \n \n \n \n \n Risk/benefit assessment, advantages over other drugs and targeting methods in the use of deferiprone as a pharmaceutical antioxidant in iron loading and non iron loading conditions.\n \n \n \n\n\n \n Kontoghiorghes, G. J., Efstathiou, A., Kleanthous, M., Michaelides, Y., & Kolnagou, A.\n\n\n \n\n\n\n Hemoglobin, 33(5): 386–397. 2009.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kontoghiorghes_riskbenefit_2009,\n\ttitle = {Risk/benefit assessment, advantages over other drugs and targeting methods in the use of deferiprone as a pharmaceutical antioxidant in iron loading and non iron  loading conditions.},\n\tvolume = {33},\n\tissn = {1532-432X 0363-0269},\n\tdoi = {10.3109/03630260903217141},\n\tabstract = {Tissue damage caused by oxidative stress is a common characteristic of many conditions involving different major organs such as the brain, heart, liver and  kidneys. The treatment of such conditions using classical antioxidants is not in  most cases sufficient or effective because it lacks specificity and has a low  therapeutic index. Increased evidence from in vitro, in vivo and clinical studies  suggest that deferiprone (L1) can be used as a potent pharmaceutical antioxidant  by mobilizing labile iron and copper and/or inhibiting their catalytic activity  in the formation of free radicals and oxidative stress in tissue damage. The high  therapeutic index, tissue penetration, rapid iron binding and clearance of the  iron complex, and the low toxicity of L1, support its application as an  antioxidant pharmaceutical for adjuvant, alternative or main therapy, especially  in conditions where other treatments have failed. Substantial clinical  improvement and reversal in most cases of the tissue damage has been observed in  cardiomyopathy in thalassemia, diabetic nephropathy and glomerulonephritis in  kidney disease, Friedreich's Ataxia and Fanconi Anemia patients. In contrast to  L1, both deferoxamine (DFO) and deferasirox (DFRA) have major disadvantages in  their use in non iron loading conditions due to toxicity implications. Further  studies in the above and other conditions and optimization of the L1 therapy in  each individual will increase the prospects of the application and role of L1 as  a universal antioxidant pharmaceutical.},\n\tlanguage = {eng},\n\tnumber = {5},\n\tjournal = {Hemoglobin},\n\tauthor = {Kontoghiorghes, George J. and Efstathiou, Aglaia and Kleanthous, Marios and Michaelides, Yiannis and Kolnagou, Anita},\n\tyear = {2009},\n\tpmid = {19814684},\n\tnote = {Place: England},\n\tkeywords = {Animals, Antioxidants/*therapeutic use, Benzoates/therapeutic use, Chelation Therapy, Copper/metabolism, Deferasirox, Deferiprone, Deferoxamine/therapeutic use, Free Radicals/metabolism, Humans, Iron Chelating Agents/*therapeutic use, Iron Overload/complications/*drug therapy, Iron/metabolism, Oxidative Stress/*drug effects, Pyridones/*therapeutic use, Risk Assessment, Siderophores/therapeutic use, Thalassemia/*drug therapy, Triazoles/therapeutic use},\n\tpages = {386--397},\n}\n\n

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\n \n\n \n \n \n \n \n Hemoglobin variants in Cyprus.\n \n \n \n\n\n \n Kyrri, A. R., Felekis, X., Kalogerou, E., Wild, B. J., Kythreotis, L., Phylactides, M., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 33(2): 81–94. 2009.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kyrri_hemoglobin_2009,\n\ttitle = {Hemoglobin variants in {Cyprus}.},\n\tvolume = {33},\n\tissn = {1532-432X 0363-0269},\n\tdoi = {10.1080/03630260902813502},\n\tabstract = {Cyprus, located at the eastern end of the Mediterranean region, has been a place of eastern and western civilizations, and the presence of various hemoglobin (Hb)  variants can be considered a testimony to past colonizations of the island. In  this study, we report the structural Hb variants identified in the Cypriot  population (Greek Cypriots, Maronites, Armenians, and Latinos) during the  thalassemia screening of 248,000 subjects carried out at the Thalassaemia Centre,  Nicosia, Cyprus, over a period of 26 years. A sample population of 65,668 people  was used to determine the frequency and localization of several of the variants  identified in Cyprus. The localization of some of the variants in regions where  the presence of foreign people was most prevalent provides important clues to the  origin of the variants. Twelve structural variants have been identified by DNA  sequencing, nine concerning the beta-globin gene and three concerning the  alpha-globin gene. The most common beta-globin variants identified were Hb S  (0.2\\%), Hb D-Punjab (0.02\\%), and Hb Lepore-Washington-Boston (Hb Lepore-WB)  (0.03\\%); the most common alpha-globin variant was Hb Setif (0.1\\%). The presence  of some of these variants is likely to be directly linked to the history of  Cyprus, as archeological monuments have been found throughout the island which  signify the presence for many years of the Greeks, Syrians, Persians, Arabs,  Byzantines, Franks, Venetians, and Turks.},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {Hemoglobin},\n\tauthor = {Kyrri, Andreani R. and Felekis, Xenia and Kalogerou, Eleni and Wild, Barbara J. and Kythreotis, Loukas and Phylactides, Marios and Kleanthous, Marina},\n\tyear = {2009},\n\tpmid = {19373583},\n\tnote = {Place: England},\n\tkeywords = {Cyprus/epidemiology, Gene Frequency, Genetic Testing, Hemoglobins, Abnormal/*analysis/genetics, Heterozygote, Humans, Thalassemia/diagnosis/*ethnology/*genetics, alpha-Globins/analysis/*genetics, beta-Globins/analysis/*genetics},\n\tpages = {81--94},\n}\n\n

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\n \n 2008\n \n \n (4)\n \n \n

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\n \n\n \n \n \n \n \n Molecular basis of thalassemia intermedia in Iran.\n \n \n \n\n\n \n Akbari, M. T., Izadi, P., Izadyar, M., Kyriacou, K., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 32(5): 462–470. 2008.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{akbari_molecular_2008,\n\ttitle = {Molecular basis of thalassemia intermedia in {Iran}.},\n\tvolume = {32},\n\tissn = {1532-432X 0363-0269},\n\tdoi = {10.1080/03630260802341851},\n\tabstract = {Thalassemia intermedia shows considerable heterogenity in phenotype and molecular basis. The aim of this study was to evaluate the prevalence and effect of  different beta-globin mutations, alpha-globin defects and (G)gamma XmnI  polymorphisms in Iranian patients. Forty-five Iranian patients with clinical  criteria of thalassemia intermedia were studied. The molecular background of the  diseases was investigated. The mean age of onset varied from 1.5 to 30 years.  Only 22.2\\% of cases received occasional blood transfusions. The hemoglobin (Hb)  level in more than half of the cases was stable (10-12 g/dL) with no need for  blood transfusions. In most cases (88.9\\%) the Hb F level was more than 50\\%.  Sixty-eight point nine percent of patients were homozygous for  beta(0)-thalassemia (beta-thal) mutations. The positive XmnI polymorphism with  the capability of enhancing Hb F production was seen in 60\\% of the studied  chromosomes. Co-inheritance of alpha-globin gene defects was seen in 22.2\\% of  cases. Only 8.9\\% of patients had beta(+) or beta(++) mutations. We concluded that  the main molecular basis of the thalassemia intermedia phenotype in Iranian cases  is co-inheritance of a positive XmnI polymorphism with beta-globin mutations,  which can enhance the capability of Hb F production. Co-inheritance of  alpha-globin defects and mild beta-globin mutations are second and third causes  of thalassemia intermedia phenotypes respectively. These factors must be  considered in genetic counseling, prediction of disease prognosis and treatment  and prenatal diagnosis.},\n\tlanguage = {eng},\n\tnumber = {5},\n\tjournal = {Hemoglobin},\n\tauthor = {Akbari, Mohammad T. and Izadi, Pantea and Izadyar, Mina and Kyriacou, Kyriacou and Kleanthous, Marina},\n\tyear = {2008},\n\tpmid = {18932071},\n\tnote = {Place: England},\n\tkeywords = {Adolescent, Adult, Child, Child, Preschool, Gene Frequency/genetics, Genotype, Humans, Infant, Iran/epidemiology, Middle Aged, Polymorphism, Genetic, Young Adult, alpha-Globins/*genetics, beta-Globins/*genetics, beta-Thalassemia/*epidemiology/*genetics},\n\tpages = {462--470},\n}\n\n

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\n \n\n \n \n \n \n \n Thalassemia and its relevance to personalized medicine.\n \n \n \n\n\n \n Kleanthous, M., & Phylactides, M.\n\n\n \n\n\n\n Personalized medicine, 5(2): 141–153. March 2008.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kleanthous_thalassemia_2008,\n\ttitle = {Thalassemia and its relevance to personalized medicine.},\n\tvolume = {5},\n\tissn = {1744-828X 1741-0541},\n\tdoi = {10.2217/17410541.5.2.141},\n\tabstract = {Thalassemias are the most common monogenic gene disorders in the world. Patients present with a wide variability of clinical phenotypes ranging from severe  phenotype (β-thalassemia major) to a very mild, almost symptomless, condition.  This variability is owing to the presence of a large number of genetic modifiers  affecting the disease. Patients are treated with blood transfusions and iron  chelation therapy. Pharmacological therapies have varying degrees of success  depending on the genetic modifiers of the disease present in the patients.  Studies undertaken to identify all the modifiers that affect β-thalassemia will  lead to more appropriate genetic counseling during prenatal diagnosis and enable  targeted and personalized treatment regimens for patients in the future.},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {Personalized medicine},\n\tauthor = {Kleanthous, Marina and Phylactides, Marios},\n\tmonth = mar,\n\tyear = {2008},\n\tpmid = {29783353},\n\tnote = {Place: England},\n\tkeywords = {gene modifiers, personalized medicine, phenotype/genotype correlations, β-thalassemia},\n\tpages = {141--153},\n}\n\n

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\n \n\n \n \n \n \n \n Hb Agrinio [alpha29(B10)Le–\\textgreateruPro (alpha2)] in combination with –(MED I). Results in a severe form of Hb H disease.\n \n \n \n\n\n \n Felekis, X., Phylactides, M., Drousiotou, A., Christou, S., Kyrri, A., Kyriakou, K., Kalogerou, E., Christopoulos, G., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 32(3): 237–246. 2008.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{felekis_hb_2008,\n\ttitle = {Hb {Agrinio} [alpha29({B10}){Le}--{\\textgreater}{uPro} (alpha2)] in combination with --({MED} {I}). {Results} in a severe form of {Hb} {H} disease.},\n\tvolume = {32},\n\tissn = {1532-432X 0363-0269},\n\tdoi = {10.1080/03630260802004103},\n\tabstract = {We report two cases of compound heterozygote patients for the --(MED I) and Hb Agrinio [alpha29(B10)Le--{\\textgreater}uPro (alpha2)] anomalies in two unrelated Greek Cypriot  families. The first patient had a serious form of Hb H disease and died at the  age of 21 due to complications arising during an operation. The second patient  showed a severe hematological picture and has been regularly transfused since an  early age. This patient exhibits bone abnormalities as well as  hepatosplenomegaly. The severity of these two incidences emphasizes the need for  the inclusion of a screening test for the --(MED I)/alpha(Agrinio)alpha genotype  among those already offered during prenatal diagnosis. Two homozygotes, as well  as a number of simple, compound, and double heterozygotes for Hb Agrinio have  been identified in Cyprus and their hematological indices are presented.},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {Hemoglobin},\n\tauthor = {Felekis, Xenia and Phylactides, Marios and Drousiotou, Anthi and Christou, Sotiroulla and Kyrri, Andreani and Kyriakou, Kyriakos and Kalogerou, Eleni and Christopoulos, George and Kleanthous, Marina},\n\tyear = {2008},\n\tpmid = {18473239},\n\tnote = {Place: England},\n\tkeywords = {*Amino Acid Substitution, *Mutation, Missense, Adult, Cyprus, Genotype, Hemoglobins, Abnormal/*genetics, Heterozygote, Humans, Male, Severity of Illness Index, Thalassemia/*genetics},\n\tpages = {237--246},\n}\n\n

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\n \n\n \n \n \n \n \n Arrayed primer extension for the noninvasive prenatal diagnosis of beta-thalassemia based on detection of single nucleotide polymorphisms.\n \n \n \n\n\n \n Papasavva, T., Kalikas, I., Kyrri, A., & Kleanthous, M.\n\n\n \n\n\n\n Annals of the New York Academy of Sciences, 1137: 302–308. August 2008.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{papasavva_arrayed_2008,\n\ttitle = {Arrayed primer extension for the noninvasive prenatal diagnosis of beta-thalassemia based on detection of single nucleotide polymorphisms.},\n\tvolume = {1137},\n\tissn = {1749-6632 0077-8923},\n\tdoi = {10.1196/annals.1448.029},\n\tabstract = {beta-Thalassemia is one of the most common autosomal recessive single-gene disorders in Cyprus. Development of a noninvasive prenatal diagnostic (NIPD)  assay for beta-thalassemia is based mostly on the detection of paternally  inherited single nucleotide polymorphisms (SNPs) using the arrayed primer  extension (APEX) method. Eleven SNPs with high degree of heterozygosity in the  Cypriot population were selected and analyzed on 34 families and the informative  SNPs were determined. The APEX assay was used on maternal plasma of seven  families using the informative SNPs; paternal allele of the fetus was  noninvasively detected in five families.},\n\tlanguage = {eng},\n\tjournal = {Annals of the New York Academy of Sciences},\n\tauthor = {Papasavva, Thessalia and Kalikas, Ioannis and Kyrri, Andreanni and Kleanthous, Marina},\n\tmonth = aug,\n\tyear = {2008},\n\tpmid = {18837964},\n\tnote = {Place: United States},\n\tkeywords = {*DNA Primers/genetics, *Polymorphism, Single Nucleotide, *beta-Thalassemia/diagnosis/genetics, Cyprus, DNA Mutational Analysis/methods, DNA/blood/genetics, Female, Fetus/physiology, Globins/genetics, Humans, Male, Pedigree, Prenatal Diagnosis/*methods},\n\tpages = {302--308},\n}\n\n

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\n \n 2006\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n Noninvasive prenatal diagnostic assay for the detection of beta-thalassemia.\n \n \n \n\n\n \n Papasavva, T., Kalakoutis, G., Kalikas, I., Neokli, E., Papacharalambous, S., Kyrri, A., & Kleanthous, M.\n\n\n \n\n\n\n Annals of the New York Academy of Sciences, 1075: 148–153. September 2006.\n Place: United States\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{papasavva_noninvasive_2006,\n\ttitle = {Noninvasive prenatal diagnostic assay for the detection of beta-thalassemia.},\n\tvolume = {1075},\n\tissn = {0077-8923},\n\tdoi = {10.1196/annals.1368.020},\n\tabstract = {The development of a noninvasive method for detection of beta-thalassemia in the population of Cyprus is based on the detection of paternally inherited single  nucleotide polymorphisms (SNPs) as well as beta-thalassemia (beta-thal)  mutations. We selected 11 informative SNPs for the Cypriot population linked to  the beta-globin locus. Two different approaches were used: allele-specific  polymerase chain reaction (AS-PCR) and the arrayed primer extension (APEX)  method. The AS-PCR approach is being standardized, and the method was applied in  two families. The paternally inherited allele was noninvasively detected with the  AS-PCR approach on maternal plasma. Some preliminary tests were performed with  the APEX method on genomic DNA of parents carrying the beta-thal mutation.},\n\tlanguage = {eng},\n\tjournal = {Annals of the New York Academy of Sciences},\n\tauthor = {Papasavva, Thessalia and Kalakoutis, Gabriel and Kalikas, Ioannis and Neokli, Electra and Papacharalambous, Soteroula and Kyrri, Andreanni and Kleanthous, Marina},\n\tmonth = sep,\n\tyear = {2006},\n\tpmid = {17108205},\n\tnote = {Place: United States},\n\tkeywords = {*Polymorphism, Single Nucleotide, *beta-Thalassemia/diagnosis/genetics, Alleles, Cyprus, Female, Humans, Infant, Newborn, Male, Pedigree, Polymerase Chain Reaction/*methods, Pregnancy, Prenatal Diagnosis/*methods, Sensitivity and Specificity},\n\tpages = {148--153},\n}\n\n

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\n\n\n

\n \n\n \n \n \n \n \n Delta-thalassemia in Cyprus.\n \n \n \n\n\n \n Pavlou, E., Phylactides, M., Kyrri, A., Kalogerou, E., Makariou, C., Georgiou, I., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 30(4): 455–462. 2006.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

\n

@article{pavlou_delta-thalassemia_2006,\n\ttitle = {Delta-thalassemia in {Cyprus}.},\n\tvolume = {30},\n\tissn = {0363-0269},\n\tdoi = {10.1080/03630260600868006},\n\tabstract = {To help clarify the hematological picture of patients who may be positive for beta- and delta-globin gene mutations, the following study was carried out. Our  aim was to identify the delta-globin gene mutations found in the Greek Cypriot  population, their frequencies and the Hb A2 values associated with them.  Seventy-four samples were selected from a random sample of 5,030 individuals, and  the database of the Molecular Genetics Thalassaemia Department containing  diagnostic analyses data was also mined for relevant information. Four novel for  Cyprus delta-globin gene mutations: -30 (T--{\\textgreater}C), Hb A2-Wrens  [delta98(FG5)Val--{\\textgreater}Met, GTG--{\\textgreater}ATG], IVS-I-2 (T--{\\textgreater}C) and Hb A2-Yokoshima  [delta25(B7)Gly--{\\textgreater}Asp (GGT--{\\textgreater}GAT)] were identified. Hb A2-Yialousa  [delta27(B9)Ala--{\\textgreater}Ser, GCC--{\\textgreater}TCC], Hb A2-Yokoshima, Hb A2-Troodos  [delta116(G18)Arg--{\\textgreater}Cys, CGC--{\\textgreater}TGC], Hb A2-Pelendri [delta141(H19)Leu--{\\textgreater}Pro,  CTG--{\\textgreater}CCG], codon 4 [delta4(A1)Thr--{\\textgreater}Ile], codon 59 (-A), Hb A2-Wrens, IVS-II-897  (A--{\\textgreater}G), IVS-I-2, -55 (T--{\\textgreater}C) and -30 bring the total to 11 delta-globin alleles  found in the Greek Cypriot population. Hb A2-Yialousa is the most common mutation  followed by codon 4, with frequencies of 60.7 and 17.8\\%, respectively.Hb A2  levels above 1.9\\% have been found to indicate a significantly reduced possibility  for the presence of a delta-globin gene mutation in this population. For Hb A2  levels of 1.7 and 1.8\\% the possibility of a delta-globin gene mutation rises to  90.9\\% and reaches 100\\% for lower Hb A2 levels. The frequency of all the mutant  delta-globin chromosomes in the sample is 0.0067 and the carrier frequency is  1.26\\%.},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {Hemoglobin},\n\tauthor = {Pavlou, Eleni and Phylactides, Marios and Kyrri, Andriani and Kalogerou, Eleni and Makariou, Christiana and Georgiou, Ioannis and Kleanthous, Marina},\n\tyear = {2006},\n\tpmid = {16987800},\n\tnote = {Place: England},\n\tkeywords = {Chi-Square Distribution, Codon/genetics, Cyprus/epidemiology, DNA Mutational Analysis, Female, Genetic Carrier Screening, Genetic Testing, Genotype, Globins/*genetics, Hemoglobin A2/*analysis, Heterozygote, Humans, Male, Point Mutation/*genetics, Sequence Analysis, DNA, alpha-Thalassemia/epidemiology/*genetics, beta-Thalassemia/epidemiology/*genetics},\n\tpages = {455--462},\n}\n\n

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\n \n 2004\n \n \n (1)\n \n \n

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\n \n \n

\n \n\n \n \n \n \n \n The impact of iron overload and genotype on gonadal function in women with thalassaemia major.\n \n \n \n\n\n \n Skordis, N., Gourni, M., Kanaris, C., Toumba, M., Kleanthous, M., Karatzia, N., Pavlides, N., & Angastiniotis, M.\n\n\n \n\n\n\n Pediatric endocrinology reviews : PER, 2 Suppl 2: 292–295. December 2004.\n Place: Israel\n\n\n\n
\n\n\n\n \n\n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{skordis_impact_2004,\n\ttitle = {The impact of iron overload and genotype on gonadal function in women with thalassaemia major.},\n\tvolume = {2 Suppl 2},\n\tissn = {1565-4753},\n\tabstract = {OBJECTIVE: The purpose of this study is to evaluate the impact of chronic iron overload and genotype on gonadal function in women with thalassaemia major.  PATIENTS AND METHODS: The study population consists of 101 women aged 15-48 years  who were treated between 1981 and 1999. These women were divided into two groups  according to their genotype: [A=no modifying genetic factor and B=presence of  modifying factors], and into four groups according to their menstrual history: NM  (normal menstruation), OLM (oligomenorrhea), PA (primary amenorrhea), and SA  (secondary amenorrhea). RESULTS: Women with NM maintained eumenorrhoea for 14.62  years, whereas those with SA did so for 6.94 years. The serial values of both FSH  and LH after stimulation with GnRH were lower in women with SA and PA (p{\\textless}0.05)  compared to women with OLM and NM. The average value of the minimum, mean and  maximum ferritin levels over a period of 20 years displayed an increasing trend  from women with NM to those with SA and PA. The lower levels of ferritin in women  in Group A did not protect them from developing SA. In addition women with SA,  who belong to Group A, had a shorter duration of eumenorrhoea compared to the  ones with SA who belong to Group B. CONCLUSIONS: Although the pathogenesis of  gonadal dysfunction in thalassaemia is known to be the consequence of iron  overload, this study demonstrates that genotype acts as an independent variable,  contributing to the development of SA in thalassaemic women.},\n\tlanguage = {eng},\n\tjournal = {Pediatric endocrinology reviews : PER},\n\tauthor = {Skordis, Nicos and Gourni, Maritsa and Kanaris, Constantinos and Toumba, Meropi and Kleanthous, Marina and Karatzia, Ntina and Pavlides, Nicos and Angastiniotis, Michael},\n\tmonth = dec,\n\tyear = {2004},\n\tpmid = {16462714},\n\tnote = {Place: Israel},\n\tkeywords = {Adolescent, Adult, Amenorrhea/*etiology/genetics/metabolism, Chelation Therapy, Female, Ferritins/blood, Follicle Stimulating Hormone/blood, Genotype, Globins/genetics, Hemoglobins/metabolism, Humans, Hypothalamo-Hypophyseal System/physiopathology, Iron Overload/blood/*complications/drug therapy/genetics, Luteinizing Hormone/blood, Menstrual Cycle, Middle Aged, Mutation, Retrospective Studies, beta-Thalassemia/blood/*complications/drug therapy/genetics},\n\tpages = {292--295},\n}\n\n

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\n \n 2003\n \n \n (1)\n \n \n

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\n \n \n

\n \n\n \n \n \n \n \n Identification of alpha-thalassemia mutations in Iranian individuals with abnormal hematological indices and normal Hb A2.\n \n \n \n\n\n \n Gohari, L. H., Petrou, M., Felekis, X., Christopoulos, G., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 27(2): 129–132. May 2003.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

\n

@article{gohari_identification_2003,\n\ttitle = {Identification of alpha-thalassemia mutations in {Iranian} individuals with abnormal hematological indices and normal {Hb} {A2}.},\n\tvolume = {27},\n\tissn = {0363-0269},\n\tdoi = {10.1081/hem-120021548},\n\tlanguage = {eng},\n\tnumber = {2},\n\tjournal = {Hemoglobin},\n\tauthor = {Gohari, Ladan H. and Petrou, Miranda and Felekis, Xenia and Christopoulos, George and Kleanthous, Marina},\n\tmonth = may,\n\tyear = {2003},\n\tpmid = {12779276},\n\tnote = {Place: England},\n\tkeywords = {*Mutation, Base Sequence, DNA Primers, Female, Genotype, Hemoglobin A2/*genetics, Humans, Iran, Male, Polymerase Chain Reaction, Restriction Mapping, alpha-Thalassemia/*genetics},\n\tpages = {129--132},\n}\n\n

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\n \n 2001\n \n \n (3)\n \n \n

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\n \n\n \n \n \n \n \n Alpha-thalassaemia prenatal diagnosis by two PCR-based methods.\n \n \n \n\n\n \n Kleanthous, M., Kyriacou, K., Kyrri, A., Kalogerou, E., Vassiliades, P., Drousiotou, A., Kallikas, I., Ioannou, P., & Angastiniotis, M.\n\n\n \n\n\n\n Prenatal diagnosis, 21(5): 413–417. May 2001.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kleanthous_alpha-thalassaemia_2001,\n\ttitle = {Alpha-thalassaemia prenatal diagnosis by two {PCR}-based methods.},\n\tvolume = {21},\n\tcopyright = {Copyright 2001 John Wiley \\& Sons, Ltd.},\n\tissn = {0197-3851},\n\tdoi = {10.1002/pd.73},\n\tabstract = {In Cyprus all couples carrying alpha0-thalassaemia mutations are detected in the course of the thalassaemia carrier screening program and prenatal diagnosis is  offered to all of them. Prenatal diagnosis for alpha-thalassaemia is routinely  done by two independent molecular methods. With the first method, the mutations  of the parents are directly determined by gap-PCR and then the chorionic villus  sample (CVS) is examined for the presence of these mutations. With the other  method, a (CA)n repeat polymorphic site located between the psialpha1- and  alpha2-globin genes is used for determining the presence or absence of the normal  and mutant alleles. In the period from 1995 to 1999, molecular analysis of 46  couples in which haematological data were consistent with deletion of two  alpha-globin genes in both partners indicated that only 13 of them were actually  at risk for haemoglobin (Hb) Bart's hydrops fetalis and prenatal diagnosis was  provided in 16 pregnancies. The molecular diagnosis was possible in all cases  with the use of both gap-PCR and (CA)n repeat polymorphisms analysis. No  misdiagnosed cases for alpha-thalassaemia have been reported to date.},\n\tlanguage = {eng},\n\tnumber = {5},\n\tjournal = {Prenatal diagnosis},\n\tauthor = {Kleanthous, M. and Kyriacou, K. and Kyrri, A. and Kalogerou, E. and Vassiliades, P. and Drousiotou, A. and Kallikas, I. and Ioannou, P. and Angastiniotis, M.},\n\tmonth = may,\n\tyear = {2001},\n\tpmid = {11360286},\n\tnote = {Place: England},\n\tkeywords = {Adult, Chorionic Villi Sampling, Cyprus/epidemiology, DNA Mutational Analysis, DNA Primers/chemistry, DNA/analysis, Female, Gene Deletion, Genetic Testing/*methods, Globins/analysis/genetics, Hemoglobins, Abnormal/analysis/genetics, Humans, Hydrops Fetalis/*diagnosis/epidemiology/genetics, Male, Molecular Epidemiology, Polymerase Chain Reaction/*methods, Polymorphism, Genetic, Pregnancy, Repetitive Sequences, Nucleic Acid, Sensitivity and Specificity, alpha-Thalassemia/*diagnosis/epidemiology/genetics},\n\tpages = {413--417},\n}\n\n

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\n \n\n \n \n \n \n \n Hb Limassol [beta8(A5)Lys–\\textgreaterAsn]: a new hemoglobin variant.\n \n \n \n\n\n \n Kyrri, A., Felekis, X., Kalogerou, E., Christopoulos, G., Makariou, C., Loizidou, D., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 25(4): 421–424. November 2001.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kyrri_hb_2001,\n\ttitle = {Hb {Limassol} [beta8({A5}){Lys}--{\\textgreater}{Asn}]: a new hemoglobin variant.},\n\tvolume = {25},\n\tissn = {0363-0269},\n\tdoi = {10.1081/hem-100107880},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {Hemoglobin},\n\tauthor = {Kyrri, A. and Felekis, X. and Kalogerou, E. and Christopoulos, G. and Makariou, C. and Loizidou, D. and Kleanthous, M.},\n\tmonth = nov,\n\tyear = {2001},\n\tpmid = {11791876},\n\tnote = {Place: England},\n\tkeywords = {Amino Acid Substitution, Cyprus, DNA Mutational Analysis, Genetic Variation, Hemoglobins, Abnormal/*genetics, Humans, Male, Mass Screening, Phenotype, Point Mutation, beta-Thalassemia/*genetics},\n\tpages = {421--424},\n}\n\n

\n

\n\n\n\n

\n\n\n

\n \n\n \n \n \n \n \n A multi-center study in order to further define the molecular basis of beta-thalassemia in Thailand, Pakistan, Sri Lanka, Mauritius, Syria, and India, and to develop a simple molecular diagnostic strategy by amplification refractory mutation system-polymerase chain reaction.\n \n \n \n\n\n \n Old, J. M., Khan, S. N., Verma, I., Fucharoen, S., Kleanthous, M., Ioannou, P., Kotea, N., Fisher, C., Riazuddin, S., Saxena, R., Winichagoon, P., Kyriacou, K., Al-Quobaili, F., & Khan, B.\n\n\n \n\n\n\n Hemoglobin, 25(4): 397–407. November 2001.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{old_multi-center_2001,\n\ttitle = {A multi-center study in order to further define the molecular basis of beta-thalassemia in {Thailand}, {Pakistan}, {Sri} {Lanka}, {Mauritius}, {Syria}, and {India},  and to develop a simple molecular diagnostic strategy by amplification refractory  mutation system-polymerase chain reaction.},\n\tvolume = {25},\n\tissn = {0363-0269},\n\tdoi = {10.1081/hem-100107877},\n\tabstract = {The spectrum of the beta-thalassemia mutations of Thailand, Pakistan, India, Sri Lanka, Mauritius and Syria has been further characterized by a multi-center study  of 1,235 transfusion-dependent patients, and the mutations discovered used to  assess the fidelity of a simple diagnostic strategy. A total of 44  beta-thalassemia mutations were identified either by allele-specific  oligonucleotide hybridization, amplification with allele-specific primers, or DNA  sequencing of amplified product. The results confirm and extend earlier findings  for Thailand, Pakistan, India, Mauritius and Syria. This is the first detailed  report of the spectrum of mutations for Sri Lanka. Two novel mutations were  identified, codon 55 (-A) and IVS-I-129 (A--{\\textgreater}C), both found in Sri Lankan  patients. Two beta-thalassemia mutations were found to coexist in one beta-globin  gene: Sri Lankan patients homozygous for the beta0 codon 16 (-C) frameshift were  also homozygous for the beta+ codon 10 (C--{\\textgreater}A) mutation. Studies of Sri Lankan,  Pakistani, and Indian carriers suggest the codon 10 (C--{\\textgreater}A) mutation is just a  rare polymorphism on an ancestral allele, on which the beta0 codon 16 (-C)  mutation has arisen. Each country was found to have only a few common mutations  accounting for 70\\% or more of the beta-thalassemia alleles. A panel of primers to  diagnose the majority of the mutations by the amplification refractory mutation  system was developed, enabling a simple molecular diagnostic strategy to be  introduced for each country participating in the multi-center study.},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {Hemoglobin},\n\tauthor = {Old, J. M. and Khan, S. N. and Verma, I. and Fucharoen, S. and Kleanthous, M. and Ioannou, P. and Kotea, N. and Fisher, C. and Riazuddin, S. and Saxena, R. and Winichagoon, P. and Kyriacou, K. and Al-Quobaili, F. and Khan, B.},\n\tmonth = nov,\n\tyear = {2001},\n\tpmid = {11791873},\n\tnote = {Place: England},\n\tkeywords = {Asia/epidemiology, Base Sequence, DNA Mutational Analysis/methods, DNA Primers, Genetic Testing/*methods, Humans, International Cooperation, Mutation, Polymerase Chain Reaction/methods, beta-Thalassemia/epidemiology/*genetics},\n\tpages = {397--407},\n}\n\n

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\n \n 2000\n \n \n (2)\n \n \n

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\n \n\n \n \n \n \n \n Identification of a novel beta0-thalassemia mutation, codons 80/81 (-C), in an Iranian family.\n \n \n \n\n\n \n Feleki, X., Najmabadi, H., Karimi-Nejad, R., Christopoulos, G., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 24(4): 319–321. November 2000.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{feleki_identification_2000,\n\ttitle = {Identification of a novel beta0-thalassemia mutation, codons 80/81 (-{C}), in an {Iranian} family.},\n\tvolume = {24},\n\tissn = {0363-0269},\n\tdoi = {10.3109/03630260008993139},\n\tlanguage = {eng},\n\tnumber = {4},\n\tjournal = {Hemoglobin},\n\tauthor = {Feleki, X. and Najmabadi, H. and Karimi-Nejad, R. and Christopoulos, G. and Kleanthous, M.},\n\tmonth = nov,\n\tyear = {2000},\n\tpmid = {11186262},\n\tnote = {Place: England},\n\tkeywords = {*Mutation, Codon, Female, Globins/*genetics, Humans, Iran, Male, beta-Thalassemia/*genetics},\n\tpages = {319--321},\n}\n\n

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\n\n\n\n

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\n \n\n \n \n \n \n \n Molecular characterization of beta-thalassemia in Syria.\n \n \n \n\n\n \n Kyriacou, K., Al Quobaili, F., Pavlou, E., Christopoulos, G., Ioannou, P., & Kleanthous, M.\n\n\n \n\n\n\n Hemoglobin, 24(1): 1–13. February 2000.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{kyriacou_molecular_2000,\n\ttitle = {Molecular characterization of beta-thalassemia in {Syria}.},\n\tvolume = {24},\n\tissn = {0363-0269},\n\tdoi = {10.3109/03630260009002268},\n\tabstract = {This study concerns the determination of beta-thalassemia alleles and other hemoglobin variants in 82 patients from Syria. We have characterized 146  chromosomes and found 17 different beta-thalassemia mutations, and one  beta-globin chain variant that gives rise to the abnormal Hb S. The eight most  common beta-thalassemia mutations were the IVS-I-110 (G--{\\textgreater}A), IVS-I-1 (G--{\\textgreater}A),  codon 5 (-CT), -30 (T--{\\textgreater}A), codon 39 (C--{\\textgreater}T), IVS-I-6 (T--{\\textgreater}C), IVS-II-1 (G--{\\textgreater}A),  and codon 15 (TGG--{\\textgreater}TAG). These mutations accounted for almost 75\\% of the total  beta-thalassemia chromosomes. We identified 34 different genotypes with a high  level of homozygosity. The various beta-thalassemia mutations were characterized  using gene amplification with specific oligonucleotide primers, restriction  enzyme analysis, denaturing gradient gel electrophoresis and direct sequencing.  By combining these three approaches we were able to detect mutations in almost  90\\% of the chromosomes studied. Our findings provide a sound foundation on which  to base a preventive program for thalassemia and we believe that the data that we  present will facilitate the improvement of medical services such as carrier  screening, genetic counseling, and prenatal diagnosis. Furthermore a detailed  knowledge of the molecular pathology of beta-thalassemia will strongly improve  the prenatal diagnosis services in Syria.},\n\tlanguage = {eng},\n\tnumber = {1},\n\tjournal = {Hemoglobin},\n\tauthor = {Kyriacou, K. and Al Quobaili, F. and Pavlou, E. and Christopoulos, G. and Ioannou, P. and Kleanthous, M.},\n\tmonth = feb,\n\tyear = {2000},\n\tpmid = {10722110},\n\tnote = {Place: England},\n\tkeywords = {Adolescent, Alleles, Amino Acid Substitution, Blood Transfusion, Child, Child, Preschool, Cyprus/epidemiology/ethnology, DNA Mutational Analysis, Frameshift Mutation, Gene Frequency, Genotype, Hemoglobins, Abnormal/genetics, Heterozygote, Homozygote, Humans, Infant, Middle East/epidemiology/ethnology, Mutation, Missense, Point Mutation, Splenectomy, Syria/epidemiology/ethnology, beta-Thalassemia/blood/*epidemiology/*genetics},\n\tpages = {1--13},\n}\n\n

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\n \n 1999\n \n \n (1)\n \n \n

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\n \n\n \n \n \n \n \n Comparative in vivo expression of beta(+)-thalassemia alleles.\n \n \n \n\n\n \n Marwan, M. M., Scerri, C. A., Zarroag, S. O., Cao, A., Kyrri, A., Kalogirou, E., Kleanthous, M., Ioannou, P., Angastiniotis, M., & Felice, A. E.\n\n\n \n\n\n\n Hemoglobin, 23(3): 221–229. August 1999.\n Place: England\n\n\n\n
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@article{marwan_comparative_1999,\n\ttitle = {Comparative in vivo expression of beta(+)-thalassemia alleles.},\n\tvolume = {23},\n\tissn = {0363-0269},\n\tdoi = {10.3109/03630269909005702},\n\tabstract = {Double heterozygotes who inherit one abnormal though stable beta-globin variant in association with a molecularly identified beta(+)-thalassaemia allele provide  unique opportunities to quantify the in vivo expression of particular  beta(+)-thalassemia alleles. The globin products of the two alleles can be  separated, quantified and the output of the beta(+)-thalassaemia allele expressed  as the MCH-beta(A) in pg beta(A)-globin/beta(+)-thalassemia allele/RBC = 0.5 MCH  x Hb A\\%. In this communication we provide new quantitative data on the expression  of five mutations as follows: the beta(+)-87 (C--{\\textgreater}G) = 3.8 pg  beta(A)-globin/beta(+)-thalassemia allele/RBC (n = 1); the beta(+) IVS-I-1  (G--{\\textgreater}A) = 0.2 pg beta(A)-globin/beta(+)-thalassemia allele/RBC (n = 1); the  beta(+) IVS-I-6 (T--{\\textgreater}C) = 2.9 pg beta(A)-globin/beta(+)-thalassemia allele/RBC (n  = 7); the beta(+) IVS-I-110 (G--{\\textgreater}A) = 1.1 pg beta(A)-globin/beta(+)-thalassemia  allele/RBC (n = 13), and the beta(+) IVS-II-745 (C--{\\textgreater}G) = 1.74 pg  beta(A)-globin/beta(+)-thalassemia allele/RBC (n = 2). The values obtained are  compared with those of other beta(+)-thalassemia alleles from the literature. It  can be seen that the MCH-beta(A) value may be a correct index of thalassemia  severity useful for the correlation of genotype with phenotype, and for  understanding the effects of mutations in beta-globin genes on  pathophysiologically meaningful beta-globin gene expression.},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {Hemoglobin},\n\tauthor = {Marwan, M. M. and Scerri, C. A. and Zarroag, S. O. and Cao, A. and Kyrri, A. and Kalogirou, E. and Kleanthous, M. and Ioannou, P. and Angastiniotis, M. and Felice, A. E.},\n\tmonth = aug,\n\tyear = {1999},\n\tpmid = {10490134},\n\tnote = {Place: England},\n\tkeywords = {Adolescent, Adult, Alleles, Child, Child, Preschool, Female, Genetic Variation, Genotype, Globins/*analysis/*genetics, Hematologic Tests, Hemoglobins, Abnormal/analysis/genetics, Hemoglobins/analysis/chemistry/genetics, Heterozygote, Homozygote, Humans, Infant, Infant, Newborn, Italy/epidemiology, Libya/epidemiology, Male, Malta/epidemiology, Middle Aged, Mutation, beta-Thalassemia/*genetics},\n\tpages = {221--229},\n}\n\n

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\n \n 1995\n \n \n (1)\n \n \n

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\n \n\n \n \n \n \n \n alpha-Thalassaemia in the population of Cyprus.\n \n \n \n\n\n \n Baysal, E., Kleanthous, M., Bozkurt, G., Kyrri, A., Kalogirou, E., Angastiniotis, M., Ioannou, P., & Huisman, T. H.\n\n\n \n\n\n\n British journal of haematology, 89(3): 496–499. March 1995.\n Place: England\n\n\n\n
\n\n\n\n \n\n \n \n doi\n \n \n\n \n link\n \n \n\n bibtex\n \n\n \n \n \n abstract \n \n\n \n\n \n \n \n \n \n \n \n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n

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@article{baysal_alpha-thalassaemia_1995,\n\ttitle = {alpha-{Thalassaemia} in the population of {Cyprus}.},\n\tvolume = {89},\n\tissn = {0007-1048},\n\tdoi = {10.1111/j.1365-2141.1995.tb08354.x},\n\tabstract = {We have determined the alpha-thalassaemia (alpha-thal) determinants in 78 patients with Hb H disease from Cyprus; 25 were Turkish Cypriots and 53 were  Greek Cypriots. Four deletional and three non-deletional alpha-thal alleles were  present; the -alpha(3.7 kb) alpha-thal-2 and the --MED-I alpha-thal-1 were most  frequently seen; --MED-II and -(alpha)20.5 deletions occurred at considerably  lower frequencies. About 15\\% of all chromosomes carried a non-deletional  alpha-thal-2 allele; of these the 5 nucleotide (nt) deletion at the first  intervening sequence (IVS-I) donor splice site was present in approximately 8\\% of  all chromosomes. Two types of polyadenylation signal (poly A) mutations were  observed. No striking frequency differences were seen between Greek and Turkish  Cypriot patients. Combinations of the various types of alpha-thal resulted in  eight different forms of Hb H disease. The phenotypes were comparable except for  great variations in the level of Hb H which was highest (average approximately  22\\%) in the 12 patients with the alpha 5nt alpha/--MED-I combination. One patient  with the same form of Hb H disease but with an additional beta-thal  (IVS-I-110,G--{\\textgreater}A) heterozygosity had a most severe microcytosis and hypochromia  with {\\textless} 1\\% Hb H. Variations in the level of Hb H might correlate with the severity  of the disease, although this was not evident from the haematological data.},\n\tlanguage = {eng},\n\tnumber = {3},\n\tjournal = {British journal of haematology},\n\tauthor = {Baysal, E. and Kleanthous, M. and Bozkurt, G. and Kyrri, A. and Kalogirou, E. and Angastiniotis, M. and Ioannou, P. and Huisman, T. H.},\n\tmonth = mar,\n\tyear = {1995},\n\tpmid = {7734346},\n\tnote = {Place: England},\n\tkeywords = {*Gene Deletion, Alleles, Cyprus, Globins/*genetics, Hemoglobin H/*analysis, Humans, Mutation, Poly A/genetics, alpha-Thalassemia/blood/*genetics},\n\tpages = {496--499},\n}\n\n

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\n \n 1992\n \n \n (1)\n \n \n

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\n \n\n \n \n \n \n \n Detection of Duchenne and Becker muscular dystrophy carriers by quantitative multiplex polymerase chain reaction analysis.\n \n \n \n\n\n \n Ioannou, P., Christopoulos, G., Panayides, K., Kleanthous, M., & Middleton, L.\n\n\n \n\n\n\n Neurology, 42(9): 1783–1790. September 1992.\n Place: United States\n\n\n\n
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@article{ioannou_detection_1992,\n\ttitle = {Detection of {Duchenne} and {Becker} muscular dystrophy carriers by quantitative multiplex polymerase chain reaction analysis.},\n\tvolume = {42},\n\tissn = {0028-3878},\n\tdoi = {10.1212/wnl.42.9.1783},\n\tabstract = {We developed a method for the detection of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) carriers. The method is based on the quantitative  analysis of the products of standard multiplex polymerase chain reaction (PCR)  from 18 different exons of the dystrophin gene, and is designated "QM-PCR." We  detected deletions of one or more exons by standard multiplex PCR in DMD/BMD  patients in 14 of 18 families examined (77.7\\%). The same deletions were readily  demonstrated by QM-PCR in nine of 14 mothers (64.3\\%) and in another six of 22  possible carriers in these families. In five families where deletions were  detectable in DMD/BMD patients, the mothers did not exhibit any deletions in  their peripheral blood (35.7\\%). We obtained evidence for germinal mosaicism in at  least two of these families and confirmed carrier identification by haplotype  analysis using CA repeat polymorphisms at the 5' and 3' ends of the dystrophin  gene. Furthermore, analysis of 17 coded DNA samples from normal females and  obligatory carriers by QM-PCR showed that this technique could directly identify  carriers of deletions in any of 18 different exons of the dystrophin gene. Its  application in combination with existing techniques is expected to significantly  improve the accuracy of carrier diagnosis in many families, and it may also be  applicable to families in which pedigree and polymorphism information is  insufficient for carrier diagnosis.},\n\tlanguage = {eng},\n\tnumber = {9},\n\tjournal = {Neurology},\n\tauthor = {Ioannou, P. and Christopoulos, G. and Panayides, K. and Kleanthous, M. and Middleton, L.},\n\tmonth = sep,\n\tyear = {1992},\n\tpmid = {1513470},\n\tnote = {Place: United States},\n\tkeywords = {Exons/genetics, Female, Genetic Carrier Screening/*methods, Haplotypes, Humans, Male, Muscular Dystrophies/*genetics, Mutation, Pedigree, Polymerase Chain Reaction/*methods},\n\tpages = {1783--1790},\n}\n\n

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