The A allele of the rs759853 single nucleotide polymorphism in the AKR1B1 gene confers risk for diabetic kidney disease in patients with type 2 diabetes from a Brazilian population

ABSTRACT Objective: The AKR1B1 gene encodes an enzyme that catalyzes the reduction of glucose into sorbitol. Chronic hyperglycemia in patients with diabetes mellitus (DM) leads to increased AKR1B1 affinity for glucose and, consequently, sorbitol accumulation. Elevated sorbitol increases oxidative stress, which is one of the main pathways related to chronic complications of diabetes, including diabetic kidney disease (DKD). Accordingly, some studies have suggested the rs759853 polymorphism in the AKR1B1 gene is associated with DKD; however, findings are still contradictory. The aim was to investigate the association of the rs759853 polymorphism in the AKR1B1 gene and DKD. Materials and methods: The sample comprised 695 patients with type 2 DM (T2DM) and DKD (cases) and 310 patients with T2DM of more than 10 years' duration, but no DKD (controls). The polymorphism was genotyped by real-time PCR. Results: Allelic and genotype frequencies of this polymorphism did not differ significantly between groups. However, the A/A genotype was associated with risk for DKD after adjustment for gender, triglycerides, BMI, presence of hypertension and diabetic retinopathy, and duration of DM, under both recessive (P = 0.048) and additive (P = 0.037) inheritance models. Conclusion: Our data suggest an association between the AKR1B1 rs759853A/A genotype and risk for DKD in Brazilians T2DM patients.

The A allele of the rs759853 single nucleotide polymorphism in the AKR1B1 gene confers risk for diabetic kidney disease in patients with type 2 diabetes from a Brazilian population.

OBJECTIVE: The AKR1B1 gene encodes an enzyme that catalyzes the reduction of glucose into sorbitol. Chronic hyperglycemia in patients with diabetes mellitus (DM) leads to increased AKR1B1 affinity for glucose and, consequently, sorbitol accumulation. Elevated sorbitol increases oxidative stress, which is one of the main pathways related to chronic complications of diabetes, including diabetic kidney disease (DKD). Accordingly, some studies have suggested the rs759853 polymorphism in the AKR1B1 gene is associated with DKD; however, findings are still contradictory. The aim was to investigate the association of the rs759853 polymorphism in the AKR1B1 gene and DKD. METHODS: The sample comprised 695 patients with type 2 DM (T2DM) and DKD (cases) and 310 patients with T2DM of more than 10 years' duration, but no DKD (controls). The polymorphism was genotyped by real-time PCR. RESULTS: Allelic and genotype frequencies of this polymorphism did not differ significantly between groups. However, the A/A genotype was associated with risk for DKD after adjustment for gender, triglycerides, BMI, presence of hypertension and diabetic retinopathy, and duration of DM, under both recessive (P = 0.048) and additive (P = 0.037) inheritance models. CONCLUSION: Our data suggest an association between the AKR1B1 rs759853A/A genotype and risk for DKD in Brazilians T2DM patients.

The G Allele of the rs12050217 Polymorphism in the BDKRB1 Gene Is Associated with Protection for Diabetic Retinopathy.

Purpose: The plasma kallikrein-kinin system is activated during vascular injury caused by diabetic retinopathy (DR), being involved in hyperpermeability and inflammation. Bradykinin B1 receptor (B1R) is expressed in human retina, and its levels are increased in murine models of diabetes. Experimental studies reveal that B1R antagonists ameliorate retinal injury caused by diabetes in rodents. Thus, the aim of this study was to investigate the association between the rs12050217A/G polymorphism in the BDKRB1 gene, the gene that codifies B1R, and DR in type 2 diabetes mellitus (T2DM) patients. Methods: We analyzed 636 T2DM patients and 443 non-diabetic subjects. T2DM patients were categorized by the presence of non-proliferative DR (NPDR, n = 267), proliferative DR (PDR, n = 197), and absence of DR (n = 172). The BDKRB1 rs12050217A/G polymorphism was genotyped by real-time PCR using TaqMan MGB probes. Results: The genotype frequencies of the BDKRB1 rs12050217A/G polymorphism are in Hardy-Weinberg equilibrium and did not differ between T2DM patients and non-diabetic subjects (P > 0.05). The presence of the genotypes containing the rs12050217 G allele was less frequent in patients with PDR when compared to patients with NPDR and without DR (32.0%, 41.9%, and 43.0%, P = 0.045, respectively). Interestingly, the presence of G allele was associated with ~40% protection for PDR, which was confirmed after correction for the presence of hypertension, ethnicity, age, HDL, and gender (odds ratio = 0.616, 95% confidence interval 0.385-0.986, P = 0.043). Conclusion: For the first time, we showed that BDKRB1 rs12050217 G allele is associated with protection for the advanced stage of DR in T2DM patients; however, further studies are needed to confirm this finding.

Effect of the g.98535683A>G SNP in the CAST gene on meat traits of Nellore beef cattle (Bos indicus) and their crosses with Bos taurus.

The objective of this study was to estimate allele frequencies of the g.98535683A>G:BTAU7 SNP in the CAST gene in different genetic groups of beef cattle produced in Brazil (Nellore and their crosses with Bos taurus), and to evaluate associations between this polymorphism and meat traits. Five hundred animals from six different genetic groups were genotyped and phenotyped for shear force (SF), myofibrillar fragmentation index (MFI), rib eye area, backfat thickness, and total lipids. Alleles A and G of the SNP were detected in all genetic groups and the frequency of A was higher than G. Significant association (P<0.05) was observed between the polymorphism and meat tenderness (SF and MFI), in which genotype AA exhibited the best values. These results demonstrate for the first time the occurrence of the studied SNP in a Zebu breed and its potential application to the genetic improvement of meat tenderness in the Nellore breed (Bos indicus) and its crosses with Bos taurus.

Marcadores moleculares aplicados no melhoramento genético de bovinos / Marcadores moleculares aplicados al mejoramiento genético de bovinos / Molecular markers applied to cattle breeding

O recente desenvolvimento da biologia molecular tem aberto a possibilidade de identificar genes importantes e utilizar a variação genômica no melhoramento genético. Os marcadores moleculares baseados em DNA atuam como ferramentas versáteis e podem ser utilizados em diferentes estudos. Desde o seu desenvolvimento, os marcadores moleculares estão sendo modificados constantemente para melhorar sua empregabilidade e trazer a possibilidade de automação do processo em análises genômicas. A descoberta da reação em cadeia da polimerase (PCR) foi um marco nesse esforço e diversos marcadores surgiram a partir do desenvolvimento da PCR. Nessa revisão, diferentes marcadores moleculares baseados em DNA desenvolvidos durante as últimas décadas foram abordados, no qual auxiliará na compreenssão das diferentes classes de marcadores genéticos existentes e suas aplicações na seleção de genótipos superiores nos programas de melhoramento genético de bovinos.

The recent development in molecular biology has opened the possibility of identifying major genes and using genomic variation in breeding programs. DNA-based molecular markers have acted as versatile tools and can be used in different studies. Since their development, molecular markers are constantly being modified to improve their employment and bring the possibility of process automation in genomic analysis. The discovery of polymerase chain reaction (PCR) was a milestone in this effort and several markers have emerged from this technique. In this review, different DNA-based molecular markers developed during the last decades have been approached, which shall aid in the understanding of the different classes of genetic markers available and their application in the selection of superior genotypes in breeding cattle.

El reciente desarrollo de la biología molecular ha abierto la posibilidad de identificar genes importantes y utilizar la variación genómica en el mejoramiento genético. Los marcadores moleculares basados en ADN actúan como herramientas versátiles y pueden ser utilizados en diferentes estudios. Desde su desarrollo, los marcadores moleculares están siendo modificados constantemente para mejorar su empleabilidad y traer la posibilidad de automatización del proceso en análisis genómicas. El descubrimiento de la reacción en cadena de polimerasa (PCR) fue un hito en ese esfuerzo y diversos marcadores han surgido a partir del desarrollo de la PCR. En esa revisión, diferentes marcadores moleculares basados en ADN desarrollados durante las últimas décadas han sido abordados, lo que ayudará en la comprensión de las distintas clases de marcadores genéticos existentes y sus aplicaciones en la selección de genotipos superiores en los programas de mejoramiento genético de bovinos.

Marcadores moleculares aplicados no melhoramento genético de bovinos / Molecular markers applied to cattle breeding / Marcadores moleculares aplicados al mejoramento genético de bovinos

O recente desenvolvimento da biologia molecular tem aberto a possibilidade de identificar genes importantes e utilizar a variação genômica no melhoramento genético. Os marcadores moleculares baseados em DNA atuam como fer ramentas versáteis e podem ser utilizados em diferentes estudos. Desde o seu desenvolvimento, os marcadores moleculares estão sendo modificados constantemente para melhorar sua empregabilidade e trazer a possibilidade de automação do processo em análises genômicas. A descoberta da reação em cadeia da polimerase (PCR) foi um marco nesse esforço e diversos marcadores surgiram a partir do desenvolvimento da PCR. Nessa revisão, diferentes marcadores moleculares baseados em DNA desenvolvidos durante as últimas décadas foram abordados, no qual auxiliará na compreenssão das diferentes classes de marcadores genéticos existentes e suas aplicações na seleção de genótipos superiores nos programas de melhoramento genético de bovinos.(AU)

The recent development in molecular biology has opened the possibility of identifying major genes and using genomic variation in breeding programs. DNA-based molecular markers have acted as versatile tools and can be used in different studies. Since their development, molecular markers are constantly being modified to improve their employment and bring the possibility of process automation in genomic analysis. The discovery of polymerase chain reaction (PCR) was a milestone in this effort and several markers have emerged from this technique. In this review, different DNA-based molecular markers developed during the last decades have been approached, which shall aid in the understanding of the different classes of genetic markers available and their application in the selection of superior genotypes in breeding cattle.(AU)

l reciente desarrollo de la biología molecular ha abierto la posibilidad de identificar genes importantes y utilizar la variación genómica en el mejoramiento genético. Los marcadores moleculares basados en ADN actúan como herramientas versátiles y pueden ser utilizados en diferentes estudios. Desde su desarrollo, los marcadores moleculares están siendo modificados constantemente para mejorar su empleabilidad y traer la posibilidad de automatización del proceso en análisis genómicas. El descubrimiento de la reacción en cadena de polimerasa (PCR) fue un hito en ese esfuerzo y diversos marcadores han surgido a partir del desarrollo de la PCR. En esa revisión, diferentes marcadores moleculares basados en ADN desarrollados durante las últimas décadas han sido abordados, lo que ayudará en la comprensión de las distintas clases de marcadores genéticos existentes y sus aplicaciones en la selección de genotipos superiores en los programas de mejora - miento genético de bovinos.(AU)

Biochemical and genetic analysis of butyrylcholinesterase (BChE) in a family, due to prolonged neuromuscular blockade after the use of succinylcholine.

Butyrylcholinesterase (BChE) is a plasma enzyme that catalyzes the hydrolysis of choline esters, including the muscle-relaxant succinylcholine and mivacurium. Patients who present sustained neuromuscular blockade after using succinylcholine usually carry BChE variants with reduced enzyme activity or an acquired BChE deficiency. We report here the molecular basis of the BCHE gene underlying the slow catabolism of succinylcholine in a patient who underwent endoscopic nasal surgery. We measured the enzyme activity of BChE and extracted genomic DNA in order to study the promoter region and all exons of the BCHE gene of the patient, her parents and siblings. PCR products were sequenced and compared with reference sequences from GenBank. We detected that the patient and one of her brothers have two homozygous mutations: nt1615 GCA > ACA (Ala539Thr), responsible for the K variant, and nt209 GAT > GGT (Asp70Gly), which produces the atypical variant A. Her parents and two of her brothers were found to be heterozygous for the AK allele, and another brother is homozygous for the normal allele. Sequence analysis of exon 1 including 5'UTR showed that the proband and her brother are homozygous for -116GG. The AK/AK genotype is considered the most frequent in hereditary hypocholinesterasemia (44%). This work demonstrates the importance of defining the phenotype and genotype of the BCHE gene in patients who are subjected to neuromuscular block by succinylcholine, because of the risk of prolonged neuromuscular paralysis.

The association of two single nucleotide polymorphisms (SNPs) in growth hormone (GH) gene with litter size and superovulation response in goat-breeds.

Two active mutations (A 781 G and A 1575 G) in growth hormone (GH) gene, and their associations with litter size (LS), were investigated in both a high prolificacy (Matou, n = 182) and a low prolificacy breed (Boer, n = 352) by using the PCR-RFLP method. Superovulation experiments were designed in 57 dams, in order to evaluate the effect of different genotypes of the GH gene on superovulation response. Two genotypes (AA and AB, CC and CD) in each mutation were detected in these two goat breeds. Neither BB nor DD homozygous genotypes were observed. The genotypic frequencies of AB and CC were significantly higher than those of AA and CD. In the third parity, Matou dams with AB or CC genotypes had significantly larger litter sizes than those with AA and CD (p < 0.05). On combining the two loci, both Matou and Boer dams with ABCD genotype had the largest litter sizes when compared to the other genotypes (p < 0.05). When undergoing like superovulation treatments, a significantly higher number of corpora lutea and ova, with a lower incidence of ovarian cysts, were harvested in the AB and CC genotypes than in AA and CD. These results show that the two loci of GH gene are highly associated with abundant prolificacy and superovulation response in goat breeds.

Exploitation of mitochondrial nad6 as a complementary marker for studying population variability in Lepidoptera.

The applicability of mitochondrial nad6 sequences to studies of DNA and population variability in Lepidoptera was tested in four species of economically important moths and one of wild butterflies. The genetic information so obtained was compared to that of cox1 sequences for two species of Lepidoptera. nad6 primers appropriately amplified all the tested DNA targets, the generated data proving to be as informative and suitable in recovering population structures as that of cox1. The proposal is that, to obtain more robust results, this mitochondrial region can be complementarily used with other molecular sequences in studies of low level phylogeny and population genetics in Lepidoptera.

Exploitation of mitochondrial nad6 as a complementary marker for studying population variability in Lepidoptera

The applicability of mitochondrial nad6 sequences to studies of DNA and population variability in Lepidoptera was tested in four species of economically important moths and one of wild butterflies. The genetic information so obtained was compared to that of cox1 sequences for two species of Lepidoptera. nad6 primers appropriately amplified all the tested DNA targets, the generated data proving to be as informative and suitable in recovering population structures as that of cox1. The proposal is that, to obtain more robust results, this mitochondrial region can be complementarily used with other molecular sequences in studies of low level phylogeny and population genetics in Lepidoptera.

Genetic variability of the fungus Beauveria spp. isolates and virulence to the lesser mealworm Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae) / Variabilidade genética de isolados de Beauveria spp. e virulência ao cascudinho Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae)

The genetic variability of 13 Beauveria spp. isolates was assessed by RAPD technique and verified the relationship between the molecular profiles generated and geographical location of isolates, the original host and pathogenicity to the lesser-mealworm Alphitobius diaperinus. Pathogenicity bioassays were carried with larvae and adult insects at fungal concentration of 109 conidia / mL, during a ten-day period. With RAPD technique, seven of 50 tested primers showed consistent banding patterns. Two groups with a similarity level above 60% were observed in the analysis profiles and one of them formed only by isolates from the lesser mealworm, with 72% of similarity. Was found relationship between the origin host of the isolates and the molecular profile, generated in the analysis. There was no relationship between the molecular profiles and the geographical location or pathogenicity to the insect. The results showed that even the isolates collected in near areas may have highly variability.

A variabilidade genética de 13 isolados de Beauveria spp. foi avaliada por meio da técnica de RAPD, para verificar se há relação entre o perfil molecular e a origem geográfica dos isolados, hospedeiro inicial e virulência ao cascudinho dos aviários (Alphitobius diaperinus). Os bioensaios de virulência foram realizados com larvas e adultos do inseto, utilizaram-se os fungos na concentração de 109 conídios ml-1 e a mortalidade dos insetos foi avaliada durante dez dias. Com a técnica de RAPD foram testados 50 primers, dos quais sete apresentaram padrões de bandas consistentes. Pela análise dos perfis gerados observou-se a separação em dois grupos com nível de similaridade superior a 60%, sendo que em um deles agruparam-se somente os isolados de cascudinho, com similaridade de 72%. Verificou-se relação entre o hospedeiro original dos isolados avaliados e o perfil molecular gerado nas análises. Não foi constatada qualquer relação entre os perfis moleculares e a região geográfica de procedência ou virulência ao inseto. Os resultados demonstraram que os isolados coletados em regiões próximas podem apresentar grande variabilidade genética.

Genetic variability of the fungus Beauveria spp. isolates and virulence to the lesser mealworm Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae) / Variabilidade genética de isolados de Beauveria spp. e virulência ao cascudinho Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae)

The genetic variability of 13 Beauveria spp. isolates was assessed by RAPD technique and verified the relationship between the molecular profiles generated and geographical location of isolates, the original host and pathogenicity to the lesser-mealworm Alphitobius diaperinus. Pathogenicity bioassays were carried with larvae and adult insects at fungal concentration of 109 conidia / mL, during a ten-day period. With RAPD technique, seven of 50 tested primers showed consistent banding patterns. Two groups with a similarity level above 60% were observed in the analysis profiles and one of them formed only by isolates from the lesser mealworm, with 72% of similarity. Was found relationship between the origin host of the isolates and the molecular profile, generated in the analysis. There was no relationship between the molecular profiles and the geographical location or pathogenicity to the insect. The results showed that even the isolates collected in near areas may have highly variability.

A variabilidade genética de 13 isolados de Beauveria spp. foi avaliada por meio da técnica de RAPD, para verificar se há relação entre o perfil molecular e a origem geográfica dos isolados, hospedeiro inicial e virulência ao cascudinho dos aviários (Alphitobius diaperinus). Os bioensaios de virulência foram realizados com larvas e adultos do inseto, utilizaram-se os fungos na concentração de 109 conídios ml-1 e a mortalidade dos insetos foi avaliada durante dez dias. Com a técnica de RAPD foram testados 50 primers, dos quais sete apresentaram padrões de bandas consistentes. Pela análise dos perfis gerados observou-se a separação em dois grupos com nível de similaridade superior a 60%, sendo que em um deles agruparam-se somente os isolados de cascudinho, com similaridade de 72%. Verificou-se relação entre o hospedeiro original dos isolados avaliados e o perfil molecular gerado nas análises. Não foi constatada qualquer relação entre os perfis moleculares e a região geográfica de procedência ou virulência ao inseto. Os resultados demonstraram que os isolados coletados em regiões próximas podem apresentar grande variabilidade genética.

Biochemical and genetic analysis of butyrylcholinesterase (BChE) in a family, due to prolonged neuromuscular blockade after the use of succinylcholine

Butyrylcholinesterase (BChE) is a plasma enzyme that catalyzes the hydrolysis of choline esters, including the muscle-relaxant succinylcholine and mivacurium. Patients who present sustained neuromuscular blockade after using succinylcholine usually carry BChE variants with reduced enzyme activity or an acquired BChE deficiency. We report here the molecular basis of the BCHE gene underlying the slow catabolism of succinylcholine in a patient who underwent endoscopic nasal surgery. We measured the enzyme activity of BChE and extracted genomic DNA in order to study the promoter region and all exons of the BCHE gene of the patient, her parents and siblings. PCR products were sequenced and compared with reference sequences from GenBank. We detected that the patient and one of her brothers have two homozygous mutations: nt1615 GCA > ACA (Ala539Thr), responsible for the K variant, and nt209 GAT > GGT (Asp70Gly), which produces the atypical variant A. Her parents and two of her brothers were found to be heterozygous for the AK allele, and another brother is homozygous for the normal allele. Sequence analysis of exon 1 including 5'UTR showed that the proband and her brother are homozygous for -116GG. The AK/AK genotype is considered the most frequent in hereditary hypocholinesterasemia (44 percent). This work demonstrates the importance of defining the phenotype and genotype of the BCHE gene in patients who are subjected to neuromuscular block by succinylcholine, because of the risk of prolonged neuromuscular paralysis.

The association of two single nucleotide polymorphisms (SNPs) in growth hormone (GH) gene with litter size and superovulation response in goat-breeds

Two active mutations (A 781 G and A 1575 G) in growth hormone (GH) gene, and their associations with litter size (LS), were investigated in both a high prolificacy (Matou, n = 182) and a low prolificacy breed (Boer, n = 352) by using the PCR-RFLP method. Superovulation experiments were designed in 57 dams, in order to evaluate the effect of different genotypes of the GH gene on superovulation response. Two genotypes (AA and AB, CC and CD) in each mutation were detected in these two goat breeds. Neither BB nor DD homozygous genotypes were observed. The genotypic frequencies of AB and CC were significantly higher than those of AA and CD. In the third parity, Matou dams with AB or CC genotypes had significantly larger litter sizes than those with AA and CD (p < 0.05). On combining the two loci, both Matou and Boer dams with ABCD genotype had the largest litter sizes when compared to the other genotypes (p < 0.05). When undergoing like superovulation treatments, a significantly higher number of corpora lutea and ova, with a lower incidence of ovarian cysts, were harvested in the AB and CC genotypes than in AA and CD. These results show that the two loci of GH gene are highly associated with abundant prolificacy and superovulation response in goat breeds.

Genetic diversity among flue-cured tobacco cultivars based on RAPD and AFLP markers

The aim of this work was to study the genetic diversity among flue-cured tobacco cultivars. RAPD and AFLP analyses were used to assess the genetic similarity among selected accessions of flue-cured tobacco. Seventy eight RAPD and 154 AFLP polymorphic bands were obtained and used to assess the genetic diversity among 28 tobacco accessions. The cultivar relationships were estimated through the cluster analysis (UPGMA) based on RAPD data and AFLP data. The accessions were grouped into three major clusters and these shared common ancestry clustered together.

Association of the heart fatty acid-binding protein gene with quality of carcass and meat traits in pigs / Associação entre o gene da proteína de ligação de ácidos graxos - coração com características de carcaça e qualidade da carne em suínos

The heart fatty acid-binding protein (HFABP) gene was sequenced in parental animals of a F2 crossing of boars of the Brazilian native Piau breed with commercial sows (Landrace x Large White Pietrain). Primers used for PCR were designed to amplify four exons of the gene. The PCR products were sequenced and compared with the GenBank sequences. Differences between the generated sequences and the GenBank sequences were observed for both genetic groups. A total of 246 F2 animals were genotyped using the Hinf I restriction enzyme. Two genotypes were identified, 198 being animals HH and 48 Hh. The Hinf I SNP was significantly associated with weights of loin (bone-in) (P<0.05), jowl (P<0.05), sirloin (P<0.10), and kidneys (P<0.01). These results showed the potential of the H-FABP gene in marker-assisted selection programs for carcass traits in pigs.(AU)

O gene da proteína de ligação de ácidos graxos - coração foi seqüenciado em animais parentais de um cruzamento F2 entre varrões da raça nativa brasileira Piau e fêmeas comerciais (Landrace x Large White x Pietrain). Os primers utilizados na reação em cadeia da polimerase foram desenhados para amplificarem os quatro éxons do gene. Os fragmentos amplificados foram seqüenciados e comparados com a seqüência do gene depositada no GenBank. Foram observadas divergências entre as seqüências geradas e as do GenBank para ambos os grupos genéticos. Foram genotipados 246 animais F2 utilizando-se a enzima Hinf I. Dois genótipos foram identificados, sendo 198 animais HH e 48 animais Hh. O polimorfismo apresentou efeito sobre o peso total do carré (P<0,05), o peso da papada (P<0,05), o peso do filezinho (P<0,10) e o peso dos rins (P<0,01). Os resultados indicam que o gene da H-FABP apresenta potencial para aplicação em programas de seleção assistida por marcadores moleculares em suínos.(AU)

Association of the heart fatty acid-binding protein gene with quality of carcass and meat traits in pigs / Associação entre o gene da proteína de ligação de ácidos graxos - coração com características de carcaça e qualidade da carne em suínos

The heart fatty acid-binding protein (HFABP) gene was sequenced in parental animals of a F2 crossing of boars of the Brazilian native Piau breed with commercial sows (Landrace x Large White Pietrain). Primers used for PCR were designed to amplify four exons of the gene. The PCR products were sequenced and compared with the GenBank sequences. Differences between the generated sequences and the GenBank sequences were observed for both genetic groups. A total of 246 F2 animals were genotyped using the Hinf I restriction enzyme. Two genotypes were identified, 198 being animals HH and 48 Hh. The Hinf I SNP was significantly associated with weights of loin (bone-in) (P<0.05), jowl (P<0.05), sirloin (P<0.10), and kidneys (P<0.01). These results showed the potential of the H-FABP gene in marker-assisted selection programs for carcass traits in pigs.

O gene da proteína de ligação de ácidos graxos - coração foi seqüenciado em animais parentais de um cruzamento F2 entre varrões da raça nativa brasileira Piau e fêmeas comerciais (Landrace x Large White x Pietrain). Os primers utilizados na reação em cadeia da polimerase foram desenhados para amplificarem os quatro éxons do gene. Os fragmentos amplificados foram seqüenciados e comparados com a seqüência do gene depositada no GenBank. Foram observadas divergências entre as seqüências geradas e as do GenBank para ambos os grupos genéticos. Foram genotipados 246 animais F2 utilizando-se a enzima Hinf I. Dois genótipos foram identificados, sendo 198 animais HH e 48 animais Hh. O polimorfismo apresentou efeito sobre o peso total do carré (P<0,05), o peso da papada (P<0,05), o peso do filezinho (P<0,10) e o peso dos rins (P<0,01). Os resultados indicam que o gene da H-FABP apresenta potencial para aplicação em programas de seleção assistida por marcadores moleculares em suínos.

ABO genotyping in leukemia patients reveals new ABO variant alleles

The ABO blood group is the most important blood group system in transfusion medicine and organ transplantation. To date, more than 160 ABO alleles have been identified by molecular investigation. Almost all ABO genotyping studies have been performed in blood donors and families and for investigation of ABO subgroups detected serologically. The aim of the present study was to perform ABO genotyping in patients with leukemia. Blood samples were collected from 108 Brazilian patients with chronic myeloid leukemia (N = 69), chronic lymphoid leukemia (N = 13), acute myeloid leukemia (N = 15), and acute lymphoid leukemia (N = 11). ABO genotyping was carried out using allele specific primer polymerase chain reaction followed by DNA sequencing. ABO*O01 was the most common allele found, followed by ABO*O22 and by ABO*A103. We identified 22 new ABO* variants in the coding region of the ABO gene in 25 individuals with leukemia (23.2%). The majority of ABO variants was detected in O alleles (15/60.0%). In 5 of 51 samples typed as blood group O (9.8%), we found non-deletional ABO*O alleles. Elucidation of the diversity of this gene in leukemia and in other diseases is important for the determination of the effect of changes in an amino acid residue on the specificity and activity of ABO glycosyltransferases and their function. In conclusion, this is the first report of a large number of patients with leukemia genotyped for ABO. The findings of this study indicate that there is a high level of recombinant activity in the ABO gene in leukemia patients, revealing new ABO variants.

The mutation G298A®Ala100Thr on th coding sequence of the Duffy antigen/chemokine receptor gene in non-caucasian Brazilians

Ala100Thr has been suggested to be a Caucasian genetic marker on the FY*B allele. As the Brazilian population has arisen from miscegenation among Portuguese, Africans, and Indians, this mutation could possibly be found in Euro- and Afro-Brazilians, or in Brazilian Indians. Fifty-three related individuals and a random sample of 100 subjects from the Brazilian population were investigated using the polymerase chain reaction and four restriction fragment length polymorphisms. Confirming the working hypothesis, among the related individuals three Afro-Brazilians (two of them a mother and daughter) and a woman of Amerindian descent had the Ala100Thr mutation on the FY*B allele. Five non-related Euro-Brazilians also carried the mutation. All nine individuals presented the Fy(a-b+) phenotype. We conclude that the Ala100Thr mutation can occur in populations other than Caucasians and that this mutation does not affect Duffy expression on red blood cells. Gene frequencies for this allele in the non-related individuals were in agreement with those of other populations. The Duffy frequencies of two Amerindian tribes were also investigated.