Assuntos
Fator VIII/genética , Hemofilia A/genética , Mosaicismo , Adulto , Saúde da Família , Feminino , Deleção de Genes , Genótipo , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Mutação , Linhagem , Fenótipo , Regiões Promotoras Genéticas , Risco , Deleção de SequênciaRESUMO
Angus steers were grazed on unsupplemented pasture (CNTRL), pasture supplemented with 0.7% BW cracked corn (FLAX-0), FLAX-0 with 0.125% and 0.250% BW of whole flaxseed (FLAX-1 and FLAX-2). Six steers were grazed per treatment for 70 days, with start and finish weights of 458 and 508 kg. At 24 h post slaughter, longissimus thoracis were harvested, and steaks assigned to treatments of postmortem aging time under vacuum (PM; 3, 14 and 56 days) with or without five days of aerobic exposure (AE). Meat antioxidant status was higher (P<0.05) when feeding CNTRL and FLAX-1 than FLAX-0 and FLAX-2. Under AE, lipid oxidation was highest for FLAX-2 (P<0.05), and lowest for FLAX-1. Greatest TBARs and lowest antioxidant capacity and redness values were obtained with AE and the longer PM (P<0.05). Beef oxidative stability through AE improved by adding a low flaxseed level to supplemented corn grain, but deteriorated by adding a high flaxseed level or by extending PM.
Assuntos
Antioxidantes/metabolismo , Dieta/veterinária , Linho/química , Peroxidação de Lipídeos , Músculo Esquelético/metabolismo , Sementes/química , Zea mays/química , Animais , Animais Endogâmicos , Antioxidantes/análise , Argentina , Bovinos , Dieta/efeitos adversos , Gorduras na Dieta/análise , Linho/efeitos adversos , Embalagem de Alimentos , Qualidade dos Alimentos , Armazenamento de Alimentos , Herbivoria , Humanos , Masculino , Carne/análise , Músculo Esquelético/crescimento & desenvolvimento , Oxirredução , Pigmentos Biológicos/análise , Pigmentos Biológicos/biossíntese , Sementes/efeitos adversos , Aumento de PesoAssuntos
Fator IX/genética , Fator VIII/genética , Hemofilia A/diagnóstico , Hemofilia A/genética , Hemofilia B/diagnóstico , Hemofilia B/genética , Reação em Cadeia da Polimerase em Tempo Real , Deleção de Sequência , Humanos , Reação em Cadeia da Polimerase em Tempo Real/economia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reprodutibilidade dos TestesRESUMO
BACKGROUND: The recessive X-linked disorder hemophilia A (HA) is rarely expressed in female carriers, most of whom express about half of normal factor VIII activity ( FVIII: C). OBJECTIVE: To propose an integrative assessment model for the binary role of the phase between the mutated F8 and the active X-chromosome (Xa) in FVIII: C in HA carriers. METHODS: We studied 67 females at risk of severe HA, comprising five symptomatic females ( FVIII: C < 1.5 IU dL(-1) ) and 14 controls. A correlation study between FVIII: C (observed vs. expected) and X-chromosome inactivation (XCI) patterns (XIPs; androgen receptor gene [AR] system) in blood leukocyte DNA was performed in carriers, by comparison of a model correlating FVIII: C and XIP with arbitrary models devoid of biological significance, and with FVIII: C levels in non-carriers (mean model) as a proxy from background data dispersion not influenced by XIP. RESULTS: We provide proof-of-concept example from a family presenting with extremely skewed XIPs in which the severe HA phenotype appeared in a heterozygous carrier of a crossover between AR and F8 loci that phased the mutated F8 with the maternally inherited Xa. Furthermore, four cases of severe HA affected women who had a combination of a heterozygous F8 mutation and extremely skewed XIPs in leukocytes or oral mucosa are presented. Correlation analyses between FVIII: C levels and XIPs in carriers (n = 38) but not in non-carriers (n = 20) showed highly significant differences between the proposed correlation model and models without biological significance. The data support a binary influence of XCI, either increasing or decreasing the FVIII: C, subject to the underlying phase set between the F8 mutation and XCI. CONCLUSIONS: Our evidence suggests that the phase between XCI and mutated F8 acts as a molecular switch conditioning FVIII: C levels and HA expression in carriers.
Assuntos
Cromossomos Humanos X , Fator VIII/genética , Hemofilia A/genética , Mutação , Inativação do Cromossomo X , Adolescente , Adulto , Estudos de Casos e Controles , Criança , Pré-Escolar , Fator VIII/análise , Feminino , Estudos de Associação Genética , Marcadores Genéticos , Predisposição Genética para Doença , Hemofilia A/sangue , Hemofilia A/diagnóstico , Hereditariedade , Heterozigoto , Humanos , Lactente , Pessoa de Meia-Idade , Linhagem , Fenótipo , Receptores Androgênicos/genética , Fatores de Risco , Índice de Gravidade de Doença , Adulto JovemRESUMO
Mutations in more than 60 different genes have been associated with non-syndromic and syndromic retinitis pigmentosa (RP), a heterogeneous group of inherited retinal dystrophies. To increase the understanding of the molecular epidemiology of the disease in Italy, we analyzed 56 patients with syndromic and non-syndromic forms of RP attending the Retinitis Pigmentosa Center of San Paolo Hospital (Milan, Italy). Patients underwent detailed clinical examination. Genomic DNA isolated from peripheral blood samples was screened for mutations in different genes according to RP form by direct sequencing analysis. The impact of novel missense mutations on protein functions was predicted by in silico analysis and protein sequence alignment. Cosegregation analysis was performed between available family members. Forty-one of the 56 probands analyzed had non-syndromic and 15 had syndromic RP forms. Putative disease-causing mutations were identified in 19 of 56 unrelated RP probands. Mutation screening identified a total of 22 different heterozygous variants. Notably, 12 of these putative pathogenic mutations have not been previously reported. New variants were found to be located on the USH2A, RPGR, EYS, and RHO genes. All 3 new variants detected in X-linked RP probands were confirmed in other affected family members. We found a positivity rate of 24.4% and 60% for probands with non-syndromic and syndromic RP, respectively. This is the first report of RPGR X-linked RP proband-ORF15 mutations in Italian patients with X-linked (XL)-RP. In addition, this is the first report of data regarding the association between EYS mutations and non-syndromic RP forms in the Italian population.
Assuntos
Predisposição Genética para Doença/genética , Mutação , Retinose Pigmentar/genética , Retinose Pigmentar/patologia , Adulto , Idoso , Sequência de Aminoácidos , Sequência de Bases , Análise Mutacional de DNA , Proteínas da Matriz Extracelular/genética , Proteínas do Olho/genética , Saúde da Família , Feminino , Humanos , Itália , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Linhagem , Rodopsina/genética , Homologia de Sequência de Aminoácidos , Síndrome , Adulto JovemRESUMO
Inhibitor development against exogenous factor VIII is a severe impairment of replacement therapy affecting 18% of Argentine patients with severe haemophilia A (HA). To study the molecular predisposition for inhibitor development, we genotyped 260 HA patients with and without inhibitors, countrywide. The inhibitor-positive population (19 transients, 15 low responders, LR and 70 high responders, HR) of 104 severe-HA patients showed 59 Inv22 (intron 22 inversions), 18 small ins/del-frameshifts, 12 gross deletions, 12 nonsense, one splicing defect and two missense, p.Arg531Pro and p.Leu575Pro, both LR and thought to impair FVIII A2 domain secondary structure. In addition, a patient with mild HA and HR showed the missense p.Glu1704Lys associated with two neutral intronic substitutions potentially affecting the A3 domain. A case/control study (84/143) permitted estimation of F8 genotype-specific inhibitor risks [OR; prevalence (CI)] in severe-HA patients classifying a high-risk group including multi-exon deletions [3.66; 55% (19-100)], Inv22 [1.8; 24% (19-100)] and nonsense in FVIII-LCh [1.2; 21% (7-59)]; an average risk group including single-exon deletions, indel frameshifts and nonsense-HCh; and a low-risk group represented by missense defects [0.14; 3% (0.6-11)]. Analysis of inhibitor concordance/discordance in related patients indicated additional genetic factors other than F8 genotype for inhibitor formation. No significant inhibitor-predisposing factors related to FVIII product exposure were found in age- and F8 genotype-stratified populations of severe-HA patients. In conclusion, the Argentine HA patient series presents similar global and mutation-specific inhibitor risks than the HA database and other published series. This case-specific information will help in designing fitted therapies and follow-up protocols in Argentina.
Assuntos
Fator VIII/antagonistas & inibidores , Fator VIII/genética , Predisposição Genética para Doença , Hemofilia A/genética , Argentina , Estudos de Casos e Controles , Humanos , Fatores de RiscoRESUMO
AIMS: To isolate and characterize bacterial strains derived from Lactobacillus casei and Lactobacillus paracasei strains and resistant to phage MLC-A. METHODS AND RESULTS: Two of nine assayed strains rendered resistant mutants with recovery efficiencies of 83% (Lact. paracasei ATCC 27092) and 100% (Lact. casei ATCC 27139). DNA similarity coefficients (RAPD-PCR) confirmed that no significant genetic changes occurred while obtaining resistant mutants. Neither parent nor mutant strains spontaneously released phages. Phage-resistant mutants were tested against phages PL-1, J-1, A2 and MLC-A8. Lactobacillus casei ATCC 27092 mutants showed, overall, lower phage resistance than Lact. paracasei ATCC 27092 ones, but still higher than that of the parent strain. Lactobacillus paracasei ATCC 27092 mutants moderately adsorbed phage MLC-A only in calcium presence, although their parent strain successfully did it with or without calcium. Adsorption rates for Lact. casei ATCC 27139 and its mutants were highly influenced by calcium. Again, phage adsorption was higher on the original strain. CONCLUSIONS: Several isolates derived from two Lact. casei and Lact. paracasei strains showed resistance to phage MLC-A but also to other Lact. casei and Lact. paracasei phages. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights isolation of spontaneous bacteriophage-resistant mutants from Lact. casei and Lact. paracasei as a good choice for use in industrial rotation schemes.
Assuntos
Bacteriófagos/fisiologia , Lacticaseibacillus casei/isolamento & purificação , Lactobacillus/isolamento & purificação , Tipagem de Bacteriófagos , Cálcio/metabolismo , DNA Bacteriano/genética , Lactobacillus/genética , Lactobacillus/virologia , Lacticaseibacillus casei/genética , Lacticaseibacillus casei/virologia , Mutação , Fenótipo , Técnica de Amplificação ao Acaso de DNA Polimórfico , Internalização do VírusRESUMO
Boar spermatozoa are sensitive to oxidative damage produced during cryopreservation. Our aim was to evaluate the participation of different antioxidants in the improvement of cryopreserved boar sperm functionality. Spermatozoa frozen with 200 µg ml(-1) α-tocopherol, 0.5 mm 17ß-oestradiol or seminal plasma were used to evaluate sperm parameters and capacitation-like changes. The 17ß-oestradiol and α-tocopherol concentrations were assessed by RIA and HPLC respectively. Motility was improved but lipid peroxidation and capacitation-like changes were diminished (P < 0.05) in antioxidant samples. A significant increase in 17ß-oestradiol concentration was detected in 17ß-oestradiol or seminal plasma samples. Alpha-tocopherol content increased in α-tocopherol, 17ß-oestradiol or seminal plasma samples, obtaining the lowest level in the α-tocopherol ones. The 17ß-oestradiol or seminal plasma components may be acting in the regeneration of the α-tocopherol antioxidant capacity. The α-tocopherol concentration may be conditioning the cryopreserved boar sperm functionality. The addition of antioxidants could be useful to reduce oxidative stress, thus improving the functionality of cryopreserved boar spermatozoa.
Assuntos
Criopreservação , Congelamento , Preservação do Sêmen , Espermatozoides/fisiologia , alfa-Tocoferol/metabolismo , Animais , Antioxidantes/metabolismo , Cromatografia Líquida de Alta Pressão , Estradiol/metabolismo , Peroxidação de Lipídeos , Masculino , Radioimunoensaio , Capacitação Espermática , Motilidade dos Espermatozoides , Espermatozoides/metabolismoRESUMO
The extraordinary heterogeneous nature of the deleterious mutations in the F8 gene that lead to functional deficiency of clotting factor VIII in haemophilia A makes routine direct mutation profiling difficult. When direct mutation analysis cannot be performed or a causative/candidate mutation is not found, a second-line approach to track the defective F8 gene within at-risk families is linkage genetic analysis with, tried-and-tested, F8-intragenic and/or extragenic non-recombining multiallelic short tandem repeats (STR). Although several typing STR loci within and around F8 have been described, there is need for improving assessment, because the combined informativeness of available assays rarely reaches 100%. Here, we characterized a newly identified 0.28 cM-resolution marker-set, consisting of a dinucleotide STR located on F8 intron 21 (F8Int21; [AC](n)) and three extragenic tetranucleotide STR located on GAB3 intron 1 (GAB3Int1; [TAAA](n)) and TMLHE intron 1 (TMLHEInt1.1; [GAAA](n) and TMLHEInt1.3; [ATTC](n)). Heterozygosity rates determined in 100 unrelated females ranged from 0.25 (GAB3Int1) to 0.63 (F8Int21). The set rendered a combined informativeness of 0.91 for at least one marker and 0.60 for a minimum of two loci, with at least one F8-intragenic. Multiallelic interlocus non-random association analysis revealed that GAB3Int1 is not in significant gametic disequilibrium (GD) with F8Int21, F8Int9.2, TMLHEInt1.3 or TMLHEInt1.1. Gametic disequilibrium breakdown attests historical recombination between GAB3Int1 and the F8 gene. Through computational analysis of reference assembly sequence data, we note in the GD breakdown region and in the F8 gene a higher than average density of the 13-mer CCNCCNTNNCCNC consensus motif, commonly associated with recombination hotspots.
Assuntos
Fator VIII/genética , Hemofilia A/genética , Íntrons/genética , Alelos , Brasil , Feminino , Frequência do Gene , Triagem de Portadores Genéticos/métodos , Marcadores Genéticos/genética , Genótipo , Humanos , Desequilíbrio de Ligação/genética , Repetições de Microssatélites , Polimorfismo GenéticoRESUMO
BACKGROUND: Inversions of F8-intron 22 (Inv22) and F8-intron 1 (Inv1) are responsible for 45-50% of severe hemophilia A cases. OBJECTIVE: In order to improve the molecular diagnosis of Inv22 and Inv1, and to enable rapid discrimination of Inv22-type 1 and Inv22-type 2 patterns, int22h-mediated deletions (Del22) and duplications (Dup22), we developed a genotyping system based on a novel inverse shifting-polymerase chain reaction (IS-PCR) approach. METHODS: IS-PCR involved BclI restriction, followed by self-ligation to create 'BclI circles', and finally PCR analysis. Three PCR analysis tests were developed: (i) Inv22-diagnostic for a pattern-sensitive detection of deleterious mutations (Inv22 and Del22) from non-deleterious variants (Dup22 and normal); (ii) Inv1-diagnostic; and (iii) Inv22-complementary for discrimination between Inv22 and Del22, and between Dup22 and normal. For rapid molecular analysis of F8, the Inv22 and Inv1 diagnostic tests can be performed simultaneously. The optional Inv22-complementary test need only be used for specific purposes. RESULTS AND CONCLUSIONS: Diagnostic tests were validated using previously studied samples. IS-PCR evaluated carrier mosaicisms and performed robustly over wide ranges of DNA qualities and procedural conditions. IS-PCR improved the molecular diagnosis of hemophilia A. This genotyping strategy may potentially be adapted to virtually all known rearrangements in the human genome.
Assuntos
Fator VIII/genética , Hemofilia A/diagnóstico , Hemofilia A/genética , Reação em Cadeia da Polimerase/métodos , Inversão Cromossômica , Análise Mutacional de DNA , Rearranjo Gênico , Genótipo , Humanos , Reação em Cadeia da Polimerase/normasRESUMO
Buffalo meat production is increasing in Argentina. Information on meat quality and nutritional value will be useful in marketing. This work describes the oxidative stability of the Longissimus dorsi (LD) in relation to consumption of antioxidant vitamins, fatty acid composition and color deterioration during ageing. Vitamins levels found in fresh beef were 4.22±0.93; 0.24±0.05 and 0.25±0.06µg/g for α- and γ-tocopherol, and ß-carotene, respectively. Vitamin loss was almost 90% throughout an ageing period of 25 days at 2°C. Concomitantly, TBARS levels increased from 0.076±0.018 to 0.14 6±0.032mg MDA/kg beef. Hexanal and pentanal levels were low and no correlations with oxidation were detected (P>0.05). The predominant color changes in aged beef were reduced redness and yellowness with an increase in lightness (P<0.05). Vitamin levels and TBARS were used to develop a prediction equation for post-mortem aging.
RESUMO
The main goal of the present work was to determine the overall antioxidant status in fresh meat from animals fed different diets and to differentiate them through their odour profiles. Attributes were evaluated in beef from pasture or grain-fed animals with (PE and GE) or without supplementation (P and G) with vitamin E (500UI/head/day). Fresh meat produced on pasture (P and PE) had higher total ferric reducing antioxidant power (FRAP) levels than meat from grain fed-animals (G and GE) (P<0.05). However, no differences were found on their ability to reduce ABTS(+) (2,2'-azino-bis-(3-ethylbenz-thiazoline-6-sulfonic acid)), indicating that total antioxidant activity was preferentially due to the reduction potential than to the quenching capacity of tissue homogenates. Two-fold glutathione (GSH and GSSG are the reduced and oxidised forms, respectively) levels were found in the P+PE group respect to G+GE meat (P<0.001). In addition, meat from pasture-fed animals presented a higher glutathione redox potential compared to grain-fed animals (-156.1±6.1 and -158.1±6.5 vs. -148.1±13 and -149.8±14.6 for P, PE G and GE, respectively), showing that the antioxidant status in fresh meat was affected by diet. Enzymatic activity of catalase and glutathione peroxidase were equivalent for all dietary groups. Only superoxide dismutase activity was slightly higher (P<0.05) in the P+PE group than in G+GE samples. Odour profile analysis was performed in relationship to antioxidant parameters. Significant linear correlation coefficients (P<0.05) were found for a set of sensors and the FRAP values. E-nose methodology successfully discriminated the odour characteristics of samples corresponding to pasture- or grain-based diet. Hence, it was possible to describe an analytical relationship between the odour profile and the antioxidant power of fresh meat.
RESUMO
PURPOSE: Was to establish the molecular genetics of thalassemias in Argentina. PATIENTS AND METHODS: Genomic DNA was amplified by PCR and six point mutations in the beta-globin gene were investigated by Dot Blot hybridization using oligonucleotide probes. The most frequent alpha-thalassemia deletions were studied by Southern Blotting. Patients were distributed in 4 groups: a) 109 beta-thalassemic carriers; b) 15 thalassemia major patients; c) 2 thalassemia intermedia patients and d) 14 probable alpha-thalassemic carriers. RESULTS: The distribution of mutated alleles in the group a) was: IVS-1 nt 1: 13.76%, IVS-1 nt 6: 7.34%, IVS-1 nt 110: 23.85%, codon 39: 39.45%, IVS-2 nt 1: 3.68% e IVS-2 nt 745: 1.83%, 10.01% could not be determined with the probes used; in the group b) the allelic distribution was similar and the compound genetic genotype were predominant related to homocygous ones; in the group c): we confirmed the presence of one beta-thalassemia mutation and a alpha gene triplication (alpha alpha alpha) in the 2 patients studied. The alpha-thalassemia character was confirmed in 8 patients of the group d) (6 had -alpha 3,7/alpha alpha genotype and 2,-alpha 3,7/-alpha 3,7 genotype). CONCLUSIONS: This study indicates that the analysis of 6 mutations in the beta-globin gene and the alpha-globin gene deletions are an effective strategy to identify thalassemias in Argentina.
Assuntos
Globinas/genética , Mutação Puntual , Talassemia/genética , Alelos , Argentina/epidemiologia , Códon/genética , Análise Mutacional de DNA , Frequência do Gene , Genótipo , Humanos , Immunoblotting , Reação em Cadeia da Polimerase , Deleção de Sequência , Talassemia/epidemiologiaRESUMO
Several investigations have presented evidence that amylin inhibits insulin secretion and induces insulin resistance both in vitro and in vivo. However, basal and postmeal amylin concentrations proved similar in non-insulin-dependent diabetes mellitus (NIDDM) patients and controls. Since hyperglycemia may alter both amylin and insulin secretion, we examined basal and glucose-stimulated amylin secretion in eight glucose-tolerant, insulin-resistant Mexican-American subjects with both parents affected with NIDDM (offspring) and correlated the findings with the insulin sensitivity data acquired by an insulin clamp. Eight offspring and eight Mexican-Americans without any family history of diabetes (controls) underwent measurement of fat free mass (3H2O dilution method), 180-minutes, 75-g oral glucose tolerance test (OGTT), and 40-mU/m2, 180-minute euglycemic insulin clamp associated with 3H-glucose infusion and indirect calorimetry. Fasting amylin was significantly increased in offspring versus controls (11.5 +/- 1.4 v 7.0 +/- 0.8 pmol/L, P < .05). After glucose ingestion, both total (3,073 +/- 257 v 1,870 +/- 202 pmol.L-1.min-1, P < .01) and incremental (1,075 +/- 170 v 518 +/- 124 pmol.L-1.min-1, P < .05) areas under the curve (AUCs) of amylin concentration were significantly greater in offspring. The amylin to insulin molar ratio was similar in offspring and controls at all time points. Basal and postglucose insulin and C-peptide concentrations were significantly increased in the offspring. No correlation was found between fasting amylin, postglucose amylin AUC or IAUC, and any measured parameter of glucose metabolism during a euglycemic-hyperinsulinemic clamp (total glucose disposal, 7.21 +/- 0.73 v 11.03 +/- 0.54, P < .001; nonoxidative glucose disposal, 3.17 +/- 0.59 v 6.33 +/- 0.56, P < .002; glucose oxidation, 4.05 +/- 0.46 v 4.71 +/- 0.21, P = NS; hepatic glucose production, 0.29 +/- 0.16 v 0.01 +/- 0.11, P = NS; all mg.min-1.kg-1 fat-free mass, offspring v controls). In conclusion, these data do not support a causal role for amylin in the genesis of insulin resistance in NIDDM.
Assuntos
Amiloide/sangue , Glicemia/metabolismo , Diabetes Mellitus Tipo 2/genética , Hiperinsulinismo/sangue , Resistência à Insulina , Insulina/sangue , Insulina/farmacologia , Adulto , Glicemia/efeitos dos fármacos , Peptídeo C/sangue , Jejum , Feminino , Glucose/metabolismo , Teste de Tolerância a Glucose , Humanos , Infusões Intravenosas , Insulina/administração & dosagem , Polipeptídeo Amiloide das Ilhotas Pancreáticas , Masculino , Americanos Mexicanos , Núcleo Familiar , Estados UnidosRESUMO
We analyzed thyroglobulin (Tg) reverse transcription polymerase chain reaction (RT-PCR) products from three congenital goiters and three normal thyroid tissues by Taq I digestion. Tg coding sequences were amplified from position 57 to 8448 in 12 amplification fragments. A Taq I restriction fragment length polymorphism was detected in the most 3' RT-PCR product (nt 7584 through 8448). Data from the sequence showed a G-->A transition (nt 7627) causing the disappearance of the Taq I site in position 7625. It produced the substitution of arginine for a glutamine at position 2510. Afterwards, we established that the glutamine allele is present in normal unrelated individuals, with an allelic frequency of 62%. This Tg variant is thus widely represented in the human population. The available sequence information from rat and bovine Tg showed the presence, in both, of glutamine at position 2510.
Assuntos
Adenina , Arginina/genética , Variação Genética , Glutamina/genética , Guanina , Tireoglobulina/genética , Alelos , Animais , Bovinos , Frequência do Gene , Bócio/genética , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , DNA Polimerase Dirigida por RNA , Ratos , Glândula Tireoide/químicaRESUMO
PURPOSE: Identification of the beta-thalassemic alleles of higher incidence in our populations. PATIENTS AND METHODS: A total of 10 families (40 subjects) were analyzed. All families consisted of one son affected by beta-thalassaemia major (10 patients) and their parents and brothers (30 subjects). Genomic DNA was extracted from peripheral blood; a segment of 536 base pairs of the human beta-globin gene was selectively amplified with oligonucleotide primers by polymerase chain reaction (PCR) and the mutations in nucleotides 1 (IVS 1-1), 6 (IVS 1-6) and 110 (IVS 1-110) of intron 1 and codon 39 of exon 2 were analyzed by hybridization with allele-specific oligonucleotide probes (ASO). RESULTS: The distribution of 20 mutated alleles of the patients was: IVS 1-1: 10%; IVS 1-6: 10%; IVS 1-110: 40%; codon 39: 30%; 2 unidentified alleles (10%) which could not be determined with the probes used in this study; 23 carriers and 1 subject with the two normal alleles, have been detected in the genetically related 30 subject population; the distribution of mutated alleles was similar to the patient with variations due to the number of children in each family. CONCLUSIONS: 1) in the analyzed population the most frequent mutations were IVS 1-110 and codon 39. -2) the diversity of beta-thalassemic alleles, together with their distribution (90% of 4 mutations), are similar to the results found in the Mediterranean area. -3) the compound genotype present in 70% of the patients is characteristic of a nonendogamic society. -4) the small number of brothers with 2 normal alleles, was noteworthy.