High Prevalence Of α-thalassemia Among Individuals With Microcytosis And Hypochromia Without Anemia

In order to determine the contribution of α-thalassemia to microcytosis and hypochromia, 339 adult outpatients seen at Unicamp University Hospital (with the exception of the Clinical Hematology outpatient clinics), who showed normal hemoglobin (Hb) levels and reduced mean corpuscular volume and mean corpuscular hemoglobin, were analyzed. Ninety-eight were Blacks (28.9%) and 241 were Caucasians (71.1%). In all cases, Hb A2 and F levels were either normal or low. The most common deletional and nondeletional forms of α-thalassemia [-α3.7, -α4.2, -MED, -(α)20.5, αHphIα, αNcolα, ααNcoI and αTSAUDI] were investigated by PCR and restriction enzyme analyses. A total of 169 individuals (49.9%) presented α-thalassemia: 145 (42.8%) were heterozygous for the -α3.7 deletion (-α3.7/αα) and 18 (5.3%) homozygous (-α3.7/-α3.7), 5 (1.5%) were heterozygous for the nondeletional form αHPhlα/αα, and 1 (0.3%) was a -MED carrier (-MED/αα). Among the Blacks, 56 (57.1%) showed the -α3.7/ αα genotype, whereas 12 (12.2%) were -α3.7/-α3.7 and I (1.0%) was an αHPhlα carrier; among the Caucasians, 89 (36.9%) were -α3.7/αα, 6 (2.5%) had the -α3.7/-α3.7 genotype, 4 (1.7%) presented the nondeletional form (αHPhlα/αα), and 1 (0.4%) was a -MED carrier. These results demonstrate that α-thalassemia, mainly through the -α3.7 deletion, is an important cause of microcytosis and hypochromia in individuals without anemia. These data are of clinical relevance since these hematological alterations are often interpreted as indicators of iron deficiency.346759762Weatherall, D.J., Clegg, J.G., (1981) The Thalassaemia Syndromes. 3rd Edn., , Blackwell Scientific Publications, OxfordBunn, H.F., Forget, B.G., (1986) Hemoglobin: Molecular, Genetics and Clinical Aspects, , W.B. Saunders, PhiladelphiaHiggs, D.R., Vickers, M.A., Wilkie, A.O.M., Pretorius, I.M., Jarman, A.P., Weatherall, D.J., A review of the molecular genetics of the human α-globin gene cluster (1989) Blood, 73, pp. 1081-1104Kazazian H., Jr., The thalassemia syndromes: Molecular basis and prenatal diagnosis in 1990 (1990) Seminars in Hematology, 27, pp. 209-228Harteveld, K.L., Losekoot, M., Ajgam, H., Van Der Wielen, M., Giordano, P.C., Bernini, L.F., α-Thalassaemia in the Netherlands: A heterogeneous spectrum of both deletions and point mutations (1997) Human Genetics, 100, pp. 465-471Higgs, D.R., α-Thalassaemia (1993) Baillieres Clinical Haematology, 6, pp. 117-150Kattamis, A.C., Camaschella, C., Sivera, P., Surrey, S., Fortina, P., Human α-thalassemia syndromes: Detection of molecular defects (1996) American Journal of Hematology, 53, pp. 81-91Bianco, I., Cappabianca, M.P., Foglietta, E., Lerone, M., Deidda, G., Morlupi, L., Grisanti, P., Graziani, B., Silent thalassemias: Genotypes and phenotypes (1997) Haematologica, 82, pp. 269-280Galanello, R., Sollaino, C., Paglietti, E., Barella, S., Perra, C., Doneddu, I., Pirroni, M.G., Cao, A., α-Thalassemia carrier identification by DNA analysis in the screening for thalassemia (1998) American Journal of Hematology, 59, pp. 273-278Sonati, M.F., Costa, F.F., Hemoglobin Bart's in a Brazilian black population (1990) Brazilian Journal of Medical and Biological Research, 23, pp. 395-396Sonati, M.F., Farah, S.B., Ramalho, A.S., Costa, F.F., High prevalence of α-thalassemia in a Black population of Brazil (1991) Hemoglobin, 15, pp. 309-311Zago, M.A., Costa, F.F., Bottura, C., Hemoglobin H disease in three Brazilian families (1984) Revista Brasileira de Genética, 7, pp. 137-147Wenning, M.R.S.C., Kimura, E.M., Costa, F.F., Saad, S.T.O., Gervásio, S.A., De Jorge, S.B., Borges, E., Sonati, M.F., α-Globin genes: Thalassemic and structural alterations in a Brazilian population (2000) Brazilian Journal of Medical and Biological Research, 33, pp. 1041-1045Dodé, C., Krishnamoorthy, R., Lamb, J., Rochette, J., Rapid analysis of -α3.7 thalassaemia and αααanti 3.7 triplication by enzymatic amplification analysis (1993) British Journal of Haematology, 82, pp. 105-111Bowden, D.K., Vickers, M.A., Higgs, D.R., A PCR-based strategy to detect the common severe determinants of a thalassaemia (1992) British Journal of Haematology, 81, pp. 104-108Oron-Karni, V., Filon, D., Oppenheim, A., Rund, D., Rapid detection of the common Mediterranean α-globin deletions/rearrangements using PCR (1998) American Journal of Hematology, 58, pp. 306-310Hall, G.W., Thein, S.L., Newland, C.A., Chisholm, J.T.S., Kanavakis, E., Kattamis, C., Higgs, D.R., A base substitution (T→C) in codon 29 of the α2-globin gene causes α thalassemia (1993) British Journal of Haematology, 85, pp. 546-552Pearson, H.A., Ehrenkranz, R.A., Rinder, H.M., Hemosiderosis in a normal child secondary to oral iron medication (2000) Pediatrics, 105, pp. 429-43

Molecular Characterization Of Hemoglobins Kurosaki [α7 Lys→glu], G-pest [α74 Asp→asn], Stanleyville-ii [α78 Asn→lys] And J-rovigo [α53 Ala→asp]

[No abstract available]1024203205Bunn, H.F., Forget, B.G., Hemoglobin: Molecular, Genetic and Clinical Aspects, p. 1986. , Philadelphia, SaundersCotton, R.G.H., (1997) Mutation Detection, , Oxford, Oxford University PressDaeie, J.V., Lewis, S.M., (1995) Practical Haematology, Ed 8, , London, Churchill LivingstoneAlter, B.P., Goff, S.C., Efremov, G.D., Gravely, M.E., Huisman, T.H.J., Globin chain electrophoresis: A new approach to determination of γ G/γ A ratio of globin synthesis (1980) Br J Haematol, 44, pp. 527-534Dodé, C., Rochette, J., Krishnamoorthy, R., Locus assignment of human α-mutations by selective amplification and direct sequencing (1990) Br J Haematol, 76, pp. 275-281Thein, S.L., Hinton, J., A single and rapid method of direct sequencing using dynabeads (1991) Br J Haematol, 79, pp. 113-115Baysal, E., Huisman, T.H.J., Detection of common deletion α-thalassemia-2 determinants by PCR (1994) Am J Haematol, 46, pp. 208-213Dodé, C., Krishnamoorthy, R., Lamb, J., Rochette, J., Rapid analysis of -α 3,7 thalassaemia and ααα anti 3,7 triplication by enzymatic amplification analysis (1993) Br J Haematol, 83, pp. 105-111Harano, T., Harano, K., Imai, K., Murakami, T., Matsuhara, H., Hb Kurosaki [α7(A5) Lys → Glu]: A new α chain variant found in a Japanese woman (1995) Hemoglobin, 19, pp. 197-201Brimhall, B., Duerst, M., Hollan, S.R., Stenzel, P., Szelényl, J., Jones, R.T., Structural characterization of hemoglobin J-Buda [α61 (E10) Lys → Asn] and G-Pest [α74(EF3) Asp → Asn] (1974) Biochim Biophys Acta, 336, pp. 344-360Costa, F.F., Sonati, M.F., Zago, M.A., Hb stanleyville-II (α 278 Asn → Lys) is associated with a -α 3,7kb α-globin gene deletion (1991) Hum Genet, 86, pp. 319-320Alberti, R., Mariuzzi, G.M., Artibani, L., Bruni, E., Tentori, L., A new haemoglobin variant: J-Rovigo alpha 53 (E-2) alanine → aspartic acid (1974) Biochim Biophys Acta, 342, pp. 1-

High prevalence of alpha-thalassemia among individuals with microcytosis and hypochromia without anemia

In order to determine the contribution of alpha-thalassemia to microcytosis and hypochromia, 339 adult outpatients seen at Unicamp University Hospital (with the exception of the Clinical Hematology outpatient clinics), who showed normal hemoglobin (Hb) levels and reduced mean corpuscular volume and mean corpuscular hemoglobin, were analyzed. Ninety-eight were Blacks (28.9%) and 241 were Caucasians (71.1%). In all cases, Hb A2 and F levels were either normal or low. The most common deletional and nondeletional forms of alpha-thalassemia [-alpha3.7, -alpha4.2, --MED, -(alpha)20.5, alphaHphIalpha, alphaNcoIalpha, aaNcoI and alphaTSAUDI] were investigated by PCR and restriction enzyme analyses. A total of 169 individuals (49.9%) presented alpha-thalassemia: 145 (42.8%) were heterozygous for the -alpha3.7 deletion (-alpha3.7/aa) and 18 (5.3%) homozygous (-alpha3.7/-alpha3.7), 5 (1.5%) were heterozygous for the nondeletional form alphaHphIalpha (alphaHphIalpha/aa), and 1 (0.3%) was a --MED carrier (--MED/aa). Among the Blacks, 56 (57.1%) showed the -alpha3.7/aa genotype, whereas 12 (12.2%) were -alpha3.7/-alpha3.7 and 1 (1.0%) was an alphaHphIalpha carrier; among the Caucasians, 89 (36.9%) were -alpha3.7/aa, 6 (2.5%) had the -alpha3.7/-alpha3.7 genotype, 4 (1.7%) presented the nondeletional form (alphaHphIalpha/aa), and 1 (0.4%) was a --MED carrier. These results demonstrate that alpha-thalassemia, mainly through the -alpha3.7 deletion, is an important cause of microcytosis and hypochromia in individuals without anemia. These data are of clinical relevance since these hematological alterations are often interpreted as indicators of iron deficiency.75976

Hb Rio Claro [β34(b16)val→met]: A Novel Electrophoretically Silent Variant Found In Association With Hb Hasharon [α47(ce5)asp→his] And α- Thalassemia-2 (-α3.7)

[No abstract available]232177182Moo-Penn, W.F., Jue, D.L., Johnson, M.H., Bechtel, K.C., Patchen, L.C., Hemoglobin variants and methods used for their characterization during 7 years of screening at the Center for Disease Control (1980) Hemoglobin, 4, pp. 347-361Weatherall, D.J., Clegg, J.B., (1981) The Thalassaemia Syndromes, 3rd Edition, , Blackwell Scientific Publications, OxfordPembrey, M.E., Macwade, P., Weatherall, D.J., Reliable routine estimation of small amounts of foetal haemoglobin by alkali denaturation (1972) J. Clin. Pathol., 25, pp. 738-740Dacie, J.V., Lewis, S.M., (1995) Practical Haematology, 8th Edition, , Churchill Livingstone, LondonAlter, B.P., Goff, S.C., Efremov, G.D., Gravely, M.E., Huisman, T.H.J., Globin chain electrophoresis: A new approach to the determination of the Gγ/Aγ ratio of globin synthesis (1980) Br. J. Haematol., 44, pp. 527-534Miranda, S.R.P., Kimura, E.M., Saad, S.T.O., Costa, F.F., Identification of Hb Zürich α2β263(E7)His→Apγ by DNA analysis in a Brazilian family (1994) Hemoglobin, 18, pp. 337-341Baysal, E., Huisman, T.H.J., Detection of common deletional α-thalassemia-2 determinants by PCR (1994) Am. J. Hematol., 46, pp. 208-213Dodé, C., Rochette, J., Krishnamoorthy, R., Locus assignment of human α-globin mutations by selective amplification and direct sequencing (1990) Br. J. Haematol., 76, pp. 275-281Thein, S.L., Hinton, J., A single and rapid method of direct sequencing using Dynabeads (1991) Br. J. Haematol., 79, pp. 113-115Blouquit, Y., Braconnier, F., Cohen-Solal, M., Földi, J., Arous, N., Ankri, A., Binet, J.L., Rosa, J., Hemoglobin Pitie-Salpetriere [β34(B16)Val→Phe]. A new high oxygen affinity variant associated with familial erythrocytosis (1980) Biochim. Biophys. Acta, 624, pp. 473-47

Hemoglobin H Disease Resulting From The Association Of The - α3.7 Rightward Deletion And The (αα)mm Deletion In A Brazilian Patient

A patient with Hb H disease resulting from the association of the - α3.7 rightward deletion with the rare (αα)MM deletion, which removes the entire α-major regulatory element (MRE), is reported. This is the first description of an α-thalassemic mutation resulting from deletion of the locus-controlling sequences in the South-American population.693179181Higgs, D.R., Vickers, M.A., Wikie, A.O.M., Petrorius, I.-M., Jarman, A.P., Weatherall, D.J., A review of the molecular genetics of the human α-globin gene cluster (1989) Blood, 73, pp. 1081-1104Higgs, D.R., The Haemoglobinopathies (1993) Baillière's Clin Haematol, 6, pp. 117-150Dacie, J.V., Lewis, S.M., (1995) Practical Haematology 8th Edn., , Edinburgh: Churchill LivingstoneBowden, D.K., Vickers, M.A., Higgs, D.R., A PCR-based strategy to detect the common severe determinants of α-thalassaemia (1992) Br J Haematol, 81, pp. 104-108Dodé, C., Krishnamoorthy, R., Lamb, J., Rochette, J., Rapid analysis of α3.7 thalassaemia and ααα triplication by enzymatic amplification analysis (1993) Br J Haematol, 82, pp. 105-111Traeger-Synodinos, J., Kanavakis, E., Tzetis, M., Kattamis, A., Kattamis, C., Characterization of nondeletional α thalassemia mutations in the Greek population (1993) Am J Hematol, 44, pp. 162-167Makonkawkeyoon, L., Sanguansermsri, T., Asato, T., Nakashima, Y., Takei, H., Rapid detection of chain termination mutation in the α2 globin gene (1993) Blood, 82, pp. 3503-3504Hall, G.E., Thein, S.L., Newland, C.A., A base substitution (T → C) in codon 29 of α2-globin gene causes α-thalassaemia (1983) Br J Haematol, 85, pp. 546-552Dodé, C., Rochette, J., Krishanamoorthy, R., Locus assignment of human α-mutation by selective amplification and direct sequencing (1990) Br J Haematol, 76, pp. 275-281Romao, L., Osório-Almeida, L., Higgs, D.R., Lavinha, J., Liebhaber, S.A., α-Thalassemia resulting from deletion of regulatory sequences far upstream of the α-globin structural genes (1991) Blood, 78, pp. 1589-1595Bernet, A., Sabatier, S., Picketts, D.J., Targeted inactivation of the Major Positive Regulatory Element (HS-40) of the human α-globin locus (1995) Blood, 86, pp. 1202-1211Higgs, D.R., (2001) Molecular Mechanisms of Alpha Thalassemia. Disorders of Hemoglobin - Genetics, Pathophysiology and Clinical Management, Chap. 17, pp. 405-430. , (Steinberg MH, Forget BG, Higgs DR, Nagel RL, eds). New York: Cambridge University Pres

Pip4kiia And β-globin: Transcripts Differentially Expressed In Reticulocytes And Associated With High Levels Of Hb H In Two Siblings With Hb H Disease

We are reporting here the results of differential gene expression experiments comparing two siblings, a 21-yr-old male and a 19-yr-old female, with the same alpha-thalassemia genotype (-α 3.7/ -SEA) and quite different levels of Hb H in the peripheral blood (18.7 and 5%, respectively). By using mRNA differential-display reverse-transcription-PCR and suppression subtractive hybridization, two main transcripts were selected in both procedures and validated by qRT-PCR, one corresponding to the phosphatidylinositol phosphate 4-kinase type II-alpha (PIP4KIIA) gene and the other to the β-globin gene, both over expressed in the patient with the higher percentage of Hb H. Type II PIP kinases produce phosphatidylinositol 4,5 biphosphate, a critical and pleiotropic regulatory molecule involved in diverse cellular activities, including gene expression. Our results suggest that PIP4KIIA may be one of the factors related to the regulation of the β-globin gene expression and the different levels of Hb H in α-thalassemic patients. © 2009 John Wiley & Sons A/S.835490493Weatherall, D.J., Clegg, J.B., (2001) The Thalassemia Syndromes, , Oxford. Blackwell ScienceChui, D.H.K., Furcharoen, S., Chan, V., Hemoglobin H disease: Not necessarily a benign disorder (2003) Blood, 101, pp. 791-800Goossens, M., Kan, Y.M., DNA analysis of hemoglobin disorders (1981) Methods Enzymol, 76, p. 805Andrade, T.G., Peterson, K.R., Cunha, A.F., Identification of novel candidate genes for globin regulation in erythroid cells containing large deletions of the human beta-globin cluster (2006) Blood Cells Mol Dis, 37, pp. 82-90Liang, P., Pardee, A.B., Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction (1992) Science, 257, pp. 967-971Altschul, S.F., Madden, T.L., Schaffer, A.A., Gapped BLAST and PSI-BLAST: A new generation of protein database search programs (1997) Nucleic Acids Res, 25, pp. 3389-3402Diatchenko, L., Lau, Y.F., Campbell, A.P., Supression subtractive hybridization: A method for generating differentially regulated or tissue-specific cDNA probes and libraries (1996) Proc Natl Acad Sci USA, 93, pp. 6025-6030Vandesompele, J., De Preter, K., Pattyn, F., Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes (2002) Genome Biol, 3, pp. 1-12Divecha, N., Irvine, R.F., Phospholipid signalling (1995) Cell, 80, pp. 269-278Anderson, R.A., Boronenkov, I.V., Doughman, S.D., Phosphatidylinositol Phosphate Kinases, a Multifaceted Family of Signaling Enzymes (1999) J Biol Chem, 274, pp. 9907-9910Doughman, R.L., Firestone, A.J., Anderson, R.A., Phosphatidilinositol phosphate kinases put P4,5(2) in its place (2003) J Membr Biol, 194, pp. 77-89Heck, J.N., Mellman, D.L., Ling, K., A conspicuous connection: Structure defines function for the phosphatidylinositol-phosphate kinase family (2007) Crit Rev Biochem Mol Biol, 42, pp. 15-39Hinchliffe, K.A., Irvine, R.F., Regulation of type II PIP kinase by PKD phosphorylation (2006) Cell Signal, 18, pp. 1906-1913Wilcox, A., Hinchliffe, K.A., Regulation of extranuclear PtIns5P production by phosphatidylinositol phosphate 4-kinase 2α (2008) FEBS Lett, 582, pp. 1391-139

Hb Campinas [α26(b7)ala → Val]: A Novel, Electrophoretically Silent, Variant

[No abstract available]242143148Bunn, H.F., Forget, B.G., (1986) Hemoglobin: Molecular, Genetic and Clinical Aspects, , W.B. Saunders Company, Philadelphia, PA, USAMoo-Penn, W.F., Jue, D.L., Johnson, M.H., Bechtel, K.C., Patchen, L.C., Hemoglobin variants and methods used for their characterization during 7 years of screening at the Center for Disease Control (1980) Hemoglobin, 4, pp. 347-361Dacie, J.V., Lewis, S.M., (1995) Practical Haematology, 8 th Edition, , Churchill Livingstone, London, EnglandAlter, B.P., Goff, S.C., Efremov, G.D., Gravely, M.E., Huisman, T.H.J., Globin chain electrophoresis: A new approach to the determination of the Gγ/ Aγ ratio of globin synthesis (1980) Br. J. Haematol., 44, pp. 527-534Pembrey, M.E., MacWade, P., Weatherall, D.J., Reliable routine estimation of small amounts of foetal haemoglobin by alkali denaturation (1972) J. Clin. Pathol., 25, pp. 738-740Blouquit, Y., Braconnier, F., Cohen-Solal, M., Foldi, J., Arous, N., Ankri, A., Binet, J.L., Rosa, J., Hemoglobin Pitie-Salpetriere [β34 (B16) Val → Phe] a new high oxygen affinity variant associated with familial erythrocytosis (1980) Biochim. Biophys. Acta, 624, pp. 473-478Dodé, C., Rochette, J., Krishnamoorthy, R., Locus assignment of human α globin mutations by selective amplification and direct sequencing (1990) Br. J. Haematol., 76, pp. 275-281Sanger, F., Nickelen, S., Coulson, A.R., DNA sequencing with chain terminating inhibitors (1977) Proc. Natl. Acad Sci. USA, 74, pp. 5463-5467Thein, S.L., Hinton, J., A simple and rapid method of direct sequencing using Dynabeads (1991) Br. J. Haematol., 79, pp. 113-115Baysal, E., Huisman, T.H.J., Detection of common deletional α-thalassemia-2 determinants by PCR (1994) Am. J. Haematol., 46, pp. 208-213Dodé, C., Krishnamoorthy, R., Lamb, J., Rochette, J., Rapid analysis of -α 3.7 thalassaemia and ααα anti 3.7 triplication by enzymatic amplification analysis (1992) Br. J. Haematol., 82, pp. 105-111Zeng, Y.-T., Huang, S.-Z., Zhou, X.-D., Qui, X.-K., Dong, Q.-Y., Li, M.-Y., Bai, J.-H., Hb Shenyang [α26 (B7) Ala → Glu]: A new unstable variant found in China (1982) Hemoglobin, 6, pp. 625-628Lacerra, G., De Angioletti, M., Di Girolamo, R., Sciorio, A., Testa, R., Schilirò, G., Carestia, C., Hb Caserta and Hb Bronte: Two novel hemoglobin variants caused by alpha-2 globin gene mutation (1997) The 6th International Conference on Thalassaemia and the Haemoglobinopathies, , Abstract 151, Malta, Apri