Resumo Várias razões podem estar subjacentes ao aumento dramático da diabetes mellitus tipo 2. Um desses motivos é a base genética e variações. Os polimorfismos do receptor da vitamina D estão associados a diferentes doenças, como artrite reumatoide e diabetes. O objetivo deste estudo é investigar a possível associação de duas mutações identificadas ApaI (rs7975232) e TaqI (rs731236). Oitenta e nove indivíduos saudáveis e 56 pacientes com diabetes tipo 2 (T2D) foram investigados usando a técnica RFLP para genotipagem e haplotipagem também. A distribuição dos genótipos Apal não foi estatisticamente significativa entre o controle (P = 0,65), bem como para os pacientes diabéticos (P = 0,58). Para as frequências do alelo Taql, o alelo T foi de 0,61, onde o alelo G foi de 0,39. A distribuição de frequência dos genótipos Taql não foi estatisticamente significativa entre o controle (P = 0,26), bem como os pacientes diabéticos (P = 0,17). O risco relativo do alelo T do gene Apa1 é 1,28 e a razão de chances do mesmo alelo é 1,53, enquanto ambas as estimativas foram < 1,0 do alelo G. Da mesma forma, com o gene Taq1, os valores de risco relativo e razão de chances para o alelo T são 1,09 e 1,27, respectivamente, e ambas as estimativas do alelo C foram de 0,86 para o risco relativo e 0,79 para o odds ratio. O desequilíbrio de ligação par a par entre os dois SNPs Taq1 / apa1 foi estatisticamente significativo no grupo de controle (D = 0,218, D' = 0,925 e valor P < 0,001) e dados semelhantes em grupos diabéticos (D = 0,2, D' = 0,875 e valor P < 0,001). Esses dados sugerem que o alelo T de ambos os genes Apa1 e Taq1 está associado ao aumento do risco de diabetes tipo 2. Achamos que precisamos de um número maior de voluntários para chegar a uma conclusão mais precisa.

Abstract Several reasons may underlie the dramatic increase in type2 diabetes mellitus. One of these reasons is the genetic basis and variations. Vitamin D receptor polymorphisms are associated with different diseases such as rheumatoid arthritis and diabetes. The aim of this study is to investigate the possible association of two identified mutations ApaI (rs7975232) and TaqI (rs731236). Eighty-nine healthy individuals and Fifty-six Type 2 Diabetic (T2D) patients were investigated using RFLP technique for genotyping and haplotyping as well. The distribution of Apal genotypes was not statistically significant among the control (P=0.65) as well as for diabetic patients (P=0.58). For Taql allele frequencies of T allele was 0.61 where of G allele was 0.39. The frequency distribution of Taql genotypes was not statistically significant among the control (P=0.26) as well as diabetic patients (P=0.17). Relative risk of the allele T of Apa1 gene is 1.28 and the odds ratio of the same allele is 1.53, while both estimates were < 1.0 of the allele G. Similarly, with the Taq1 gene the relative risk and the odds ratio values for the allele T are 1.09 and 1.27 respectively and both estimates of the allele C were 0.86 for the relative risk and 0.79 for the odds ratio. The pairwise linkage disequilibrium between the two SNPs Taq1/apa1 was statistically significant in control group (D = 0.218, D' = 0.925 and P value < 0.001) and similar data in diabetic groups (D = 0.2, D' = 0.875 and P value < 0.001). These data suggest that the T allele of both genes Apa1 and Taq1 is associated with the increased risk of type 2 diabetes. We think that we need a larger number of volunteers to reach a more accurate conclusion.

Resumo O amarelecimento letal (LY) é uma doença que afecta o coqueiro e outras espécies de palmeiras. Está associada a fitoplasmas do grupo 16SrIV e o único vector de insectos notificado até agora para este patogéneo é o Haplaxius crudus. O H. crudus está presente no México e foi associado aos fitoplasmas do grupo 16SrIV, no entanto, não foi detectável durante um surto de LY na costa de Yucatan, México, sugerindo a existência de outras espécies de vectores. Para testar esta hipótese foi realizado um levantamento de insectos e um total de 3074 insectos foram capturados durante um ano de amostragem mensal. Dez ordens taxonómicas foram identificadas nesta amostra, sendo Hemiptera a mais abundante (N=2094), e estas foram classificadas em nove famílias. A colpoptera sp. da família Nogodinidae era a mais abundante, representando 56% do número total de insectos amostrados e 23% destas amostras resultaram positivos para o fitoplasma LY por detecção por PCR. A comparação BLAST, RFLP virtual e análises filogenéticas dos amplicons sequenciados relacionam o fitoplasma detectado com o subgrupo 16SrIV-A. Os resultados aqui apresentados sugerem que Colpoptera sp. poderia ser considerado como um novo vector putativo dos fitoplasmas causadores de LY no México e um candidato a investigação adicional.

Abstract Lethal yellowing (LY) is a disease that affects coconut and other palm species. It is associated to phytoplasmas of the group 16SrIV and the only reported insect vector for this pathogen so far is Haplaxius crudus. H. crudus is present in Mexico and has been associated to 16SrIV phytoplasmas, however, it was not detectable during a LY outbreak in the coast of Yucatan, Mexico, suggesting the existence of other vector species. To test this hypothesis a survey of insects was carried out and a total of 3074 insects were captured during a year of monthly sampling. Ten taxonomic orders were identified in this sample, Hemiptera being the most abundant (N=2094), and these were classified into nine families. The leafhopper Colpoptera sp. from to the Nogodinidae family was de most abundant representing 56% of the total number of insects sampled and 23% of these samples resulted positive for LY phytoplasma by PCR detection. The BLAST comparison, virtual RFLP and phylogenetic analyses of the sequenced amplicons relate the detected phytoplasma to the subgroup 16SrIV-A. The findings presented herein suggest that Colpoptera sp. could be considered as a new putative vector of the LY-causing phytoplasmas in Mexico and a candidate for further research.

ABSTRACT This study aimed to optimize a reliable and rapid genotyping assay to detect the carriers of bovine citrullinemia (BC), bovine leukocyte adhesion deficiency (BLAD), complex vertebral malformation (CVM), and deficiency of uridine monophosphate synthase (DUMPS) in cattle populations. We developed real-time polymerase chain reaction (RT-PCR)-based assays to distinguish wild-type and defective alleles of BLAD, CVM, BC, and DUMPS. Twenty-four bulls in the International Center for Livestock Research and Training were genotyped. At the same time, genotyping was performed for DUMPS and BC using PCR-RFLP, and sequencing was performed for all diseases and compared with the RT-PCR kit we developed. None of the bulls carried mutant alleles of these hereditary autosomal recessive lethal defects. It takes only 2 h for the assay to be completed, including DNA extraction from the sample. These consequences indicate that RT-PCR is an easy, reliable, and rapid method for detecting BLAD, CVM, BC, and DUMPS carriers. Studies show a high frequency of mutant alleles of these genetic defects that cause genetic diseases, and this requires routine test systems to eradicate genetic diseases of economic importance. , (BC) BLAD (BLAD) CVM (CVM) (DUMPS populations realtime real time RTPCRbased RT PCR based wildtype wild type Twentyfour Twenty four genotyped PCRRFLP, PCRRFLP RFLP, RFLP PCR-RFLP RTPCR completed sample easy importance (BC (BLAD (CVM

RESUMO A leishmaniose é um dos mais importantes dilemas de saúde enfrentados pela Organização Mundial da Saúde (OMS) devido à sua ampla disseminação e à grande diversidade de flebotomíneos que a transmitem. Este estudo teve como objetivo detectar a presença de parasitas de Leishmania nos flebotomíneos espalhados em campos de refugiados por meio da técnica PCR-RLFP. Um total de 437 flebotomíneos foi coletado e classificado em duas espécies: Phlebotomus papatasi e Phlebotomus sergenti. O DNA foi extraído das espécies de flebotomíneos das fêmeas e, em seguida, a reação de PCR foi amplificada por dois primers (LITSR, L5.8S) que transcreveram um gene parcial do espaçador transcrito interno (ITS)-1 para o parasita Leishmania com um comprimento de 320 pb. A PCR mostrou a presença de DNA de Leishmania em fêmeas de P. papatasi (10%) e P. sergenti (20%). Para determinar as espécies de Leishmania transmitidas pelas duas espécies de moscas anteriores, a técnica de RFLP-PCR foi realizada pela enzima HaeIII para o DNA de Leishmania extraído delas. A RFLP-PCR mostrou que as fêmeas de P. papatasi transmitiam Leishmania major e as fêmeas de P. sergenti transmitiam Leishmania tropica nos campos de refugiados. Pode-se concluir que a leishmaniose está amplamente distribuída nos campos de refugiados devido à presença de seu vetor. OMS (OMS transmitem PCRRLFP. PCRRLFP RLFP. RLFP PCR-RLFP 43 seguida LITSR, LITSR (LITSR L5.8S L58S LS L5 8S L S ITS1 ITS 1 (ITS)- 32 pb P 10% 10 (10% 20%. 20 20% . (20%) anteriores RFLPPCR RFLP delas Podese Pode se vetor 4 (ITS) 3 (10 2 (20% (ITS (1 (20 ( (2

ABSTRACT Leishmaniasis is one of the most important health dilemmas facing the World Health Organization (WHO), due to it being widespread and the great diversity of sand flies that transmit it. This study aimed to detect the presence of Leishmania parasites in the sand flies spread in Refugee camps by PCR- RLFP technique. A total of 437 sandflies were collected and classified into two species Phlebotomus papatasi and Phlebotomus sergenti. DNA was extracted from the female fly species, then the PCR reaction was amplified by two primers (LITSR, L5.8S) that transcribed a partial internal transcribed spacer (ITS)-1 gene for Leishmania parasite with a length of 320 bp. PCR showed the presence of Leishmania DNA in females of both P. papatasi (10%) and P. sergenti (20%). To determine Leishmania species transmitted by the two previous fly species, the RFLP-PCR technique was performed by the HaeIII enzyme for Leishmania DNA extracted from them. RFLP-PCR showed that P. papatasi females transmitted Leishmania major and P. sergenti females transmitted Leishmania tropica in Refugee camps. It could be concluded that leishmaniasis is widely distributed in Refugee camps due to the presence of its vector. WHO, WHO , (WHO) 43 LITSR, LITSR (LITSR L5.8S L58S LS L5 8S L S ITS1 ITS 1 (ITS)- 32 bp P 10% 10 (10% 20%. 20 20% . (20%) RFLPPCR RFLP them vector (WHO 4 (ITS) 3 (10 2 (20% (ITS (1 (20 ( (2

ABSTRACT Introduction: Hemoglobinopathy Sβ-thalassemia (HbSβ-thal) has a wide range of clinical and laboratory severity. There is limited information on the natural history of HbSβ-thal and its modulating factors. We described the molecular, hematological, and clinical characteristics of a cohort of children with HbSβ-thal and estimated its incidence in Minas Gerais, Brazil. Methods: Laboratory and clinical data were retrieved from medical records. Molecular analysis was performed by HBB gene sequencing, PCR-RFLP, gap-PCR, and MLPA. Results: Eighty-nine children were included in the study. Fourteen alleles of β-thal mutations were identified. The incidence of HbSβ-thal in the state was 1 per 22,250 newborns. The most common βS-haplotypes were CAR and Benin. The most frequent βthal-haplotypes were V, II, and I. Coexistence of 3.7 kb HBA1/HBA2 deletion was present in 21.3 % of children. β-thalassemia mutations were associated with several clinical and laboratory features. In general, the incidence of clinical events per 100 patient-years was similar for children with HbSβ0-thal, IVS-I-5 G&gt;A, and IVS-I-110 G&gt;A. Children with HbSβ+-intermediate phenotypes had a more severe laboratory and clinical profile when compared with those with HbSβ+-mild ones. βS-haplotypes and α-thalassemia did not meaningfully influence the phenotype of children with HbSβ-thal. Conclusion: The early identification of b-thalassemia alleles may help the clinical management of these children. © 2023 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier España, S.L.U. This is an open access article under the CC BY-NC-ND license Introduction Sβthalassemia Sβ thalassemia HbSβthal HbSβ thal (HbSβ-thal severity factors molecular hematological Gerais Brazil Methods records sequencing PCRRFLP, PCRRFLP PCR RFLP, RFLP PCR-RFLP gapPCR, gapPCR gap PCR, gap-PCR MLPA Results Eightynine Eighty nine study βthal β identified 22250 22 250 22,25 newborns βShaplotypes βS haplotypes Benin βthalhaplotypes V II I 37 3 7 3. HBA1HBA2 HBAHBA HBA1 HBA2 HBA HBA1/HBA 213 21 21. βthalassemia features general 10 patientyears patient years HbSβ0thal, HbSβ0thal HbSβ0 thal, HbSβ0-thal IVSI5 IVSI IVS 5 IVS-I- G&gtA, GgtA G&gt A, G gt A G&gt;A IVSI110 110 IVS-I-11 G&gtA. A. HbSβ+intermediate HbSβintermediate HbSβ+ intermediate HbSβ+mild HbSβmild mild ones αthalassemia α HbSβthal. thal. Conclusion bthalassemia b 202 Hematologia Celular España SLU S L U S.L.U BYNCND BY NC ND 2225 2 25 22,2 HBA1HBA IVS-I gtA G&gtA Ggt IVSI11 11 IVS-I-1 20 222 22, IVSI1

SUMMARY OBJECTIVE: Nonalcoholic fatty liver disease is a chronic liver disease and a growing global epidemic. The aim of this study was to investigate the association between a visfatin gene (NAMPT) variant and nonalcoholic fatty liver disease, owing to the connection between this disease and insulin resistance, obesity, inflammation, and oxidative stress, and the role of visfatin in these metabolic disorders. METHODS: In the present case-control study, we enrolled 312 genetically unrelated individuals, including 154 patients with biopsy-proven nonalcoholic fatty liver disease and 158 controls. The rs2058539 polymorphism of NAMPT gene was genotyped using the PCR-RFLP method. RESULTS: Genotype and allele distributions of NAMPT gene rs2058539 polymorphism conformed to the Hardy-Weinberg equilibrium both in the case and control groups (p>0.05). The distribution of NAMPT rs2058539 genotypes and alleles differed significantly between the cases with nonalcoholic fatty liver disease and controls. The "CC" genotype of the NAMPT rs2058539 compared with "AA" genotype was associated with a 2.5-fold increased risk of nonalcoholic fatty liver disease after adjustment for confounding factors [p=0.034; odds ratio (OR)=2.52, 95% confidence interval (CI)=1.36–4.37]. Moreover, the NAMPT rs2058539 "C" allele was significantly overrepresented in the nonalcoholic fatty liver disease patients than controls (p=0.022; OR=1.77, 95%CI=1.14–2.31). CONCLUSION: Our findings indicated for the first time that the NAMPT rs2058539 "CC" genotype is a marker of increased nonalcoholic fatty liver disease susceptibility; however, it needs to be supported by further investigations in other populations. OBJECTIVE epidemic (NAMPT resistance obesity inflammation stress disorders METHODS casecontrol 31 individuals 15 biopsyproven biopsy proven rs rs205853 PCRRFLP PCR RFLP method RESULTS HardyWeinberg Hardy Weinberg p>0.05. p005 p p>0.05 . 0 05 (p>0.05) CC "CC AA "AA 2.5fold 25fold fold 2.5 2 5 p=0.034 p0034 034 [p=0.034 OR=2.52, OR252 OR =2.52, 52 (OR)=2.52 95 CI=1.36–4.37. CI136437 CI =1.36–4.37 1 36 4 37 (CI)=1.36–4.37] Moreover C "C p=0.022 p0022 022 (p=0.022 OR177 77 OR=1.77 95%CI=1.14–2.31. 95CI114231 95%CI=1.14–2.31 14 95%CI=1.14–2.31) CONCLUSION susceptibility however populations 3 rs20585 p00 p>0.0 (p>0.05 5fold 25 2. p=0.03 p003 03 [p=0.03 OR=2.52 OR25 252 =2.52 (OR)=2.5 9 CI=1.36–4.37 CI13643 136437 =1.36–4.3 (CI)=1.36–4.37 p=0.02 p002 02 (p=0.02 OR17 7 OR=1.7 95CI11423 95%CI=1.14–2.3 rs2058 p0 p>0. (p>0.0 p=0.0 [p=0.0 OR=2.5 OR2 =2.5 (OR)=2. CI=1.36–4.3 CI1364 13643 =1.36–4. (CI)=1.36–4.3 (p=0.0 OR1 OR=1. 95CI1142 95%CI=1.14–2. rs205 p>0 (p>0. p=0. [p=0. OR=2. =2. (OR)=2 CI=1.36–4. CI136 1364 =1.36–4 (CI)=1.36–4. (p=0. OR=1 95CI114 95%CI=1.14–2 rs20 p> (p>0 p=0 [p=0 OR=2 =2 (OR)= CI=1.36–4 CI13 136 =1.36– (CI)=1.36–4 (p=0 OR= 95CI11 95%CI=1.14– rs2 (p> p= [p= = (OR) CI=1.36– CI1 13 =1.36 (CI)=1.36– (p= 95CI1 95%CI=1.14 (p [p (OR CI=1.36 =1.3 (CI)=1.36 95CI 95%CI=1.1 CI=1.3 =1. (CI)=1.3 95%CI=1. CI=1. =1 (CI)=1. 95%CI=1 CI=1 (CI)=1 95%CI= CI= (CI)= 95%CI (CI) (CI

SUMMARY OBJECTIVE: The aim of this study was to evaluate the vitamin D receptor (VDR) BsmI variant in morbidly obese patients compared with healthy normal controls. METHODS: The study included 103 patients with morbid obesity and 120 healthy individuals serving as normal controls. The DNA samples obtained from blood were genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The gender, age, smoking status, triglycerides, total cholesterol, insulin, mean body mass index, and frequency of allele and genotype of the BsmI variant in the VDR gene in morbidly obese patients were evaluated. RESULTS: The body mass index of the patients was 47.14 ± 7.19. The VDR B/B, B/b, and b/b genotype frequencies were 27.2% versus 28.3%; 54.4% versus 50%; and 18.4% versus 21.7% in the morbidly obese patients and the control group, respectively. There was no statistically significant difference between patients and control subjects in the genotype and allele distribution of the VDR BsmI variant (p>0.05). Both patients and control genotype frequencies are consistent with Hardy-Weinberg equilibrium. CONCLUSION: The BsmI variant in the VDR gene may not seem to predispose to morbid obesity in our study population. Further studies with a larger number of subjects are needed to make a more precise evaluation of this relationship. OBJECTIVE (VDR controls METHODS 10 12 reactionrestriction reaction restriction PCRRFLP PCR RFLP (PCR-RFLP technique gender age status triglycerides cholesterol insulin evaluated RESULTS 4714 47 14 47.1 719 7 19 7.19 BB B B/B Bb b B/b bb 272 27 2 27.2 28.3% 283 28 3 544 54 4 54.4 50% 50 184 18 18.4 217 21 21.7 group respectively p>0.05. p005 p p>0.05 . 0 05 (p>0.05) HardyWeinberg Hardy Weinberg equilibrium CONCLUSION population relationship 1 471 47. 71 7.1 27. 28.3 5 54. 18. 21. p00 p>0.0 (p>0.05 7. 28. p0 p>0. (p>0.0 p>0 (p>0. p> (p>0 (p> (p

SUMMARY OBJECTIVES: Nonalcoholic fatty liver disease is the term used for a range of conditions in which fat builds up in the liver and exceeds 5% of hepatocytes without inordinate alcohol intake or other causes of lipid accumulation. Regarding the fact that insulin resistance and obesity play key roles in the pathogenesis of nonalcoholic fatty liver disease, as well as the connection between resistin and these metabolic diseases, the association between nonalcoholic fatty liver disease and a resistin gene (RETN) polymorphism was examined. METHODS: In this genetic case–control association study, 150 biopsy-proven nonalcoholic fatty liver disease patients and 154 controls were enrolled and genotyped for the RETN rs1862513 (-420C>G) gene polymorphism using PCR–RFLP method. RESULTS: The −420C>G genotype frequency distributions in both groups were consistent with Hardy-Weinberg equilibrium (HWE; p>0.05). The carriers of the RETN −420C>G "CC" genotype compared with the "GG" genotype occurred less frequently in the cases with nonalcoholic fatty liver disease than in the controls, and the difference remained significant even after adjustment for confounding factors (p=0.030; OR=0.47, 95%CI=0.36–0.93). Interestingly, the RETN −420C>G "C" allele was also associated with a decreased risk for nonalcoholic fatty liver disease too (p=0.042; OR=0.72, 95%CI=0.53–0.95). CONCLUSION: We found for the first time an association between biopsy-proven nonalcoholic fatty liver disease and RETN −420C>G promoter polymorphism. The carriers of the RETN −420C>G "CC" genotype had a 53% decreased risk for nonalcoholic fatty liver disease. Our findings, however, need to be corroborated by further studies. OBJECTIVES 5 accumulation diseases (RETN examined METHODS casecontrol case control study 15 biopsyproven biopsy proven rs rs186251 420C>G 420CG CG 420C G C (-420C>G PCRRFLP PCR RFLP method RESULTS HardyWeinberg Hardy Weinberg HWE (HWE p>0.05. p005 p p>0.05 . 0 05 p>0.05) CC "CC GG "GG p=0.030 p0030 030 (p=0.030 OR047 OR 47 OR=0.47 95%CI=0.36–0.93. 95CI036093 CI 95%CI=0.36–0.93 95 36 93 95%CI=0.36–0.93) Interestingly "C p=0.042 p0042 042 (p=0.042 OR072 72 OR=0.72 95%CI=0.53–0.95. 95CI053095 95%CI=0.53–0.95 53 95%CI=0.53–0.95) CONCLUSION findings however studies 1 rs18625 p00 p>0.0 p=0.03 p003 03 (p=0.03 OR04 4 OR=0.4 95CI03609 95%CI=0.36–0.9 9 3 p=0.04 p004 04 (p=0.04 OR07 7 OR=0.7 95CI05309 95%CI=0.53–0.9 rs1862 p0 p>0. p=0.0 (p=0.0 OR0 OR=0. 95CI0360 95%CI=0.36–0. 95CI0530 95%CI=0.53–0. rs186 p>0 p=0. (p=0. OR=0 95CI036 95%CI=0.36–0 95CI053 95%CI=0.53–0 rs18 p> p=0 (p=0 OR= 95CI03 95%CI=0.36– 95CI05 95%CI=0.53– rs1 p= (p= 95CI0 95%CI=0.36 95%CI=0.53 (p 95CI 95%CI=0.3 95%CI=0.5 95%CI=0. 95%CI=0 95%CI= 95%CI

Abstract The aim of this study was to investigate the DGAT1 gene polymorphism and its effects on lamb weight in kazakh and tajik sheep breeds. A total of 97 blood samples were collected from purebred (еdilbay х еdilbay) and crossbred lambs (еdilbay x gissar) breеd by the Baiserke Agro Scientific and Production Center in the Talgar District of the Almaty Region of Kazakhstan. Animals were genotyped for DGAT1-AluI polymorphism using the polymerase chain reaction-restriction length polymorphism (PCR-RFLP) method. The result of PCR-RFLP showed that purebred (еdilbay х еdilbay) sheep had three genotypes (CC, CT and TT) and crossbred sheep had two genotypes (CC and CT). The predominant genotype was CC with a frequency of 0.70 and 0.58 in purebred sheep and crossbred sheep breeds, respectively. The DGAT1 gene showed no significant association with live weight of lambs at different times in both breeds studied. However, the study showed that the CC genotype produced higher live weight at day 60 in purebred sheep (CC: 33,668 kg and CT: 32,444) and at day 120 (CC: 41,487 and CT: 40,929) in crossbred lambs. The present study was the first to investigate the polymorphism and relationships between genotypes and lamb live weights for DGAT1 gene in sheep breeds, purebred and crossbred. We conclude that further comprehensive investigations should be done for the exact evidence of the effects of DGAT1/Aluı polymorphism on lamb live weights. DGAT 9 еdilbay gissar Kazakhstan DGAT1AluI DGATAluI AluI reactionrestriction reaction restriction PCRRFLP PCR RFLP (PCR-RFLP method CC, TT CT. . CT) 070 0 70 0.7 058 58 0.5 respectively studied However 6 33668 33 668 33,66 32,444 32444 32 444 12 41487 41 487 41,48 40,929 40929 40 929 DGAT1Aluı DGATAluı Aluı 07 7 0. 05 5 3366 3 66 33,6 32,44 3244 44 1 4148 4 48 41,4 40,92 4092 92 336 33, 32,4 324 414 41, 40,9 409 32, 40,

Resumo O objetivo deste estudo foi investigar o polimorfismo do gene DGAT1 e seus efeitos no peso do cordeiro em duas raças de ovelhas cazaques e tadjiques. Um total de 97 amostras de sangue foram coletadas de cordeiros de raça pura (еdilbay × еdilbay) e mestiços (еdilbay × gissar) criados pelo Baiserke Agro Scientific and Production Center no distrito de Talgar da região de Almaty do Cazaquistão. Os animais foram genotipados para o polimorfismo DGAT1-AluI usando o método de reação em cadeia da polimerase-polimorfismo de comprimento de restrição (PCR-RFLP). O resultado da PCR-RFLP mostrou que ovelhas de raça pura (еdilbay × еdilbay) tinham três genótipos (CC, CT e TT) e ovelhas mestiças tinham dois genótipos (CC e CT). O genótipo predominante foi CC com uma frequência de 0,70 e 0,58 em ovelhas de raça pura e mestiças, respectivamente. O gene DGAT1 não mostrou associação significativa com o peso vivo dos cordeiros, em diferentes momentos, em ambas as raças estudadas. No entanto, o estudo mostrou que o genótipo CC produziu maior peso vivo no dia 60 em ovelhas de raça pura (CC: 33,668 kg e CT: 32,444 kg) e, no dia 120, (CC: 41,487 kg e CT: 40,929 kg) em cordeiros mestiços. O presente estudo foi o primeiro a investigar o polimorfismo e as relações entre genótipos e pesos vivos de cordeiros para o gene DGAT1 em raças de ovelhas, puras e mestiças. Concluímos que investigações mais abrangentes devem ser feitas para a evidência exata dos efeitos do polimorfismo DGAT1/AluI nos pesos vivos dos cordeiros. DGAT tadjiques 9 еdilbay gissar Cazaquistão DGAT1AluI DGATAluI AluI polimerasepolimorfismo polimerase PCRRFLP. PCRRFLP PCR RFLP . (PCR-RFLP) CC, TT CT. CT) 070 0 70 0,7 058 58 0,5 respectivamente momentos estudadas entanto 6 33668 33 668 33,66 32444 32 444 32,44 120 41487 41 487 41,48 40929 40 929 40,92 (PCR-RFLP 07 7 0, 05 5 3366 3 66 33,6 3244 44 32,4 12 4148 4 48 41,4 4092 92 40,9 336 33, 324 32, 1 414 41, 409 40,

ABSTRACT Introduction: The Band 3 is a red blood cell protein that carries the Dia and Dib antigens from the Diego blood system. The SLC4A1 gene encodes Band 3; Band 3 Memphis is a polymorphism of normal Band 3 and has two variants, but only the variant II carries the Dia antigen. Objectives: Describe the frequencies of the DI*A and DI*B alleles and the Band 3 Memphis among blood donors, sickle cell disease (SCD) patients and Amazonian Indians. Methods: A total of 427 blood samples were collected and separated into three groups: 206 unrelated blood donors, 90 patients with SCD and 131 Amazonian Indians. We performed DI*A/B, normal Band 3 and Band 3 Memphis genotyping, using the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR–RFLP). Results: The frequency of the DI*A/DI*A genotype was 0.5% in blood donors and it was not found in other groups. The frequency of the DI*A/DI*B was higher in Amazonian Indians (33.6%) and the frequency of the DI*B/DI*B was highest in blood donors (92.2%). All 105 individuals tested were positive for the presence of normal Band 3 and of these individuals, only 5/105 (4.8%) presented the Band 3 Memphis mutation. Conclusion: We observed a higher frequency of the DI*B allele in blood donors and a low frequency of the DI*A/DI*A genotype in all groups studied. The Band 3 Memphis was found in a higher frequency in the blood donor group. Our findings highlight the importance of analyzing different population groups to gain a better understanding of the genetic association of blood group antigens. Introduction system SLCA SLC SLC4A variants antigen Objectives DIA DI DIB B (SCD Methods 42 20 9 13 DIAB DI*A/B genotyping PCR–RFLP. PCRRFLP PCR–RFLP . PCR RFLP (PCR–RFLP) Results DIADIA 05 0 5 0.5 DIADIB 33.6% 336 33 6 (33.6% DIBDIB 92.2%. 922 92.2% 92 2 (92.2%) 10 5105 5/10 4.8% 48 4 8 (4.8% mutation Conclusion studied 1 (PCR–RFLP 0. 33.6 (33.6 92.2 (92.2% 510 5/1 4.8 (4.8 33. (33. 92. (92.2 51 5/ 4. (4. (33 (92. (4 (3 (92 ( (9

Abstract Background: Vitiligo is an acquired depigmented skin disorder. It has a genetic and autoimmune background. Human beta defensin-1(HBD-1) plus its gene polymorphism were linked to some autoimmune disorders. Objectives: To elucidate the possible role of HBD-1 in the pathogenesis of non-segmental vitiligo (NSV) through evaluation of HBD-1 serum levels and its single nucleotide polymorphism (SNP) in patients having NSV, in addition, to correlating the results with the extent of vitiligo in those patients. Methods: A current case-control study included 50 patients having NSV and 50 controls. The authors used Vitiligo Area Scoring Index (VASI) score to assess vitiligo severity and laboratory investigations to assess serum HBD-1 level using ELISA and defensin-beta1 (DEFB1) SNP using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: There were significantly lower HBD-1 serum levels in NSV cases than in controls (p < 0.001). There was a significant predominance of GG DEFB1 genotype and G allele in NSV patients in comparison to controls (p < 0.001). The levels of serum HBD-1 and DEFB1 genotypes were not associated or correlated significantly with any of the personal and clinical parameters of vitiligo patients. Study limitations: The small sample size. Conclusions: DEFB1 gene polymorphism (GG genotype and G allele) may modulate vitiligo risk and contribute to vitiligo development in Egyptian populations. Decreased circulating HBD-1 levels might have an active role in vitiligo etiopathogenesis that could be mediated through its possible anti-inflammatory effects. Background disorder background defensin1HBD1 defensinHBD defensin 1 HBD defensin-1(HBD-1 disorders Objectives HBD1 HBD- nonsegmental non segmental (NSV (SNP addition Methods casecontrol case control 5 VASI (VASI defensinbeta1 defensinbeta beta1 defensin-beta DEFB (DEFB1 reactionrestriction reaction restriction PCRRFLP. PCRRFLP PCR RFLP . (PCR-RFLP) Results p 0.001. 0001 0.001 0 001 0.001) limitations size Conclusions populations antiinflammatory anti inflammatory effects defensin1HBD defensin-1(HBD- (DEFB (PCR-RFLP 000 0.00 00 defensin-1(HBD 0.0 0.

Abstract Introduction Phagocytosis of autoantibody-sensitized coated platelets through Fc gamma receptors on phagocytic cells is an important mechanism of thrombocytopenia in primary immune thrombocytopenia (ITP). Objective We aimed to investigate the contribution of the FcγRIIa and FcγRIIIa genes polymorphism to the risk of ITP and their association with disease characteristics in Egyptian children. Methods A case control study was conducted on eighty children with primary ITP and eighty age and sex healthy matched subjects as a control group. The FcγRIIa and FcγRIIIa genes polymorphism was detected using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results We found that the FcγRIIa‐131H and ‐131R allele frequencies were 51.3 % and 48.7%, respectively, in children with ITP, versus 75% and 25%, respectively, in controls (p= 0.002). The compound heterozygous HR genotype was significantly higher in ITP patients (p < 0.05). The FcγRIIIa-158F and ‐158V allele frequencies were 46.3% and 53.7%, respectively, in children with ITP, versus 70% and 30%, respectively, in controls (p= 0.002). The compound heterozygous VF genotype was significantly higher in ITP patients (p < 0.05). The combined HR/FV genotype was 47.5% in ITP patients, versus 10% in controls (p < 0.001). No significant difference was found between children with newly diagnosed ITP and those who developed chronic ITP, regarding the frequency distribution of the FcγRIIa and FcγRIIIa alleles and genotypes (p > 0.05). Conclusion There is a possible association of the FcγRIIa and FcγRIIIa genes polymorphism with the risk for, and genetic susceptibility to ITP in Egyptian children, but large-scale studies are still needed to support our findings. autoantibodysensitized autoantibody sensitized ITP. . (ITP) group reactionrestriction reaction restriction PCRRFLP. PCRRFLP PCR RFLP (PCR-RFLP) FcγRIIa131H FcγRIIaH 131H H 131R R 513 51 3 51. 487 48 7 48.7% respectively 75 25 25% p= p 0.002. 0002 0.002 0 002 0.002) 0.05. 005 0.05 05 0.05) FcγRIIIa158F FcγRIIIaF 158F F 158V V 463 46 46.3 537 53 53.7% 70 30 30% HRFV FV 475 47 5 47.5 10 0.001. 0001 0.001 001 0.001) for largescale large scale findings (ITP (PCR-RFLP 4 48.7 2 000 0.00 00 0.0 46. 53.7 47. 1 48. 0. 53.

Resumen Clostridioides difficile es un anaerobio esporulado que se asocia con episodios de diarreas hospitalarias. Su virulencia se encuentra vinculada, principalmente, a las toxinas TcdA y TcdB, codificadas por sus respectivos genes, tcdA y tcdB, que son parte de un locus de patogenicidad (PaLoc). Nuestro objetivo fue caracterizar los toxinotipos de C. difficile circulantes en un hospital público. Los genes tcdA y tcdB fueron amplificados y digeridos con diferentes enzimas de restricción: EcoRI para tcdA; HincII y AccI para tcdB. Además, se evaluó la presencia de cdtB (gen de la toxina binaria B) y de las toxinas A y B (por dot blot), así como el efecto citotóxico de sobrenadantes de cultivo sobre células Vero. En conjunto, estos estudios revelaron tres toxinotipos circulantes según la clasificación de Rupnik: 0, I y VIII; el más prevalente fue el último. Aunque son necesarios más estudios (ej., secuenciación), es interesante notar que la presencia del toxinotipo I podría estar relacionada con la introducción de bacterias de diferente origen geográfico. En los pacientes infectados con el toxinotipo VIII, el análisis multivariante de los resultados de laboratorio mostró que los aislamientos asociados a decesos (GCD13, GCD14 y GCD22) estaban situados en regiones de los biplots relacionados con valores de laboratorio alterados al momento de la internación. En los otros pacientes, aunque no se observó correlación entre los valores de laboratorio al momento de la internación y la evolución clínica, los niveles de urea, creatinina y recuento de glóbulos blancos estuvieron correlacionados positivamente entre sí una vez diagnosticada la infección. Nuestro estudio revela la circulación de diferentes toxinotipos de C. difficile en un mismo hospital público. La variedad de toxinotipos puede originarse a partir de microorganismos preexistentes en la región, así como también por la introducción de bacterias provenientes de otras regiones geográficas. La existencia de microorganismos con diferente potencial patogénico es relevante para el control, el seguimiento y el tratamiento de las infecciones.

Abstract Clostridioides difficile is a spore-forming anaerobe microorganism associated to nosocomial diarrhea. Its virulence is mainly associated with TcdA and TcdB toxins, encoded by their respective tcdA and tcdB genes. These genes are part of the pathogenicity locus (PaLoc). Our aim was to characterize relevant C. difficile toxinotypes circulating in the hospital setting. The tcdA and tcdB genes were amplified and digested with different restriction enzymes: EcoRI for tcdA; HincII and AccI for tcdB. In addition, the presence of the cdtB (binary toxin) gene, TcdA and TcdB toxins by dot blot and the cytotoxic effect of culture supernatants on Vero cells, were evaluated. Altogether, these studies revealed three different circulating toxinotypes according to Rupnik's classification: 0, I and VIII, being the latter the most prevalent one. Even though more studies are certainly necessary (e.g. sequencing analysis), it is worth noting that the occurrence of toxinotype I could be related to the introduction of bacteria from different geographical origins. The multivariate analysis conducted on the laboratory values of individuals infected with the most prevalent toxinotype (VIII) showed that the isolates associated with fatal outcomes (GCD13, GCD14 and GCD22) are located in regions of the biplots related to altered laboratory values at admission. In other patients, although laboratory values at admission were not correlated, levels of urea, creatinine and white blood cells were positively correlated after the infection was diagnosed. Our study reveals the circulation of different toxinotypes of C. difficile strains in this public hospital. The variety of toxinotypes can arise from pre-existing microorganisms as well as through the introduction of bacteria from other geographical regions. The existence of microorganisms with different pathogenic potential is relevant for the control, follow-up, and treatment of the infections.

Abstract The aim of this study was to determine the presence of deoxyribonucleic acid (DNA) from Toxoplasma gondii, Sarcocystis spp. and Neospora caninum, in tissues of wild boars slaughtered in southern Brazil. A total of 156 samples were collected from different organs of 25 wild boars, and DNA from at least one of the protozoa investigated was detected in 79 samples. To differentiate between infectious agents, restriction fragment length polymorphism was performed using the restriction enzymes DdeI and HpaII. For N. caninum, conventional PCR was performed with specific primers. The DNA of at least one of the studied pathogens was detected in each animal: 26.58% for T. gondii, 68.36% for Sarcocystis spp. and 5.06% for N. caninum. Coinfection between T. gondii and Sarcocystis spp. occurred in 14 animals, between T. gondii and N. caninum in only one male animal, between Sarcocystis spp. and N. caninum in a female, while co-infection with the three agents was equally observed in only one male animal. Considering the high frequency of detection and its zoonotic risk, especially T. gondii, it appears that wild boars can be potential sources of transmission of infectious agents and the adoption of monitoring measures in these populations should be prioritized. (DNA spp Brazil 15 2 7 HpaII N primers animal 2658 26 58 26.58 T 6836 68 36 68.36 506 5 06 5.06 1 animals female coinfection co infection risk prioritized 265 26.5 683 6 3 68.3 50 0 5.0 26. 68. 5.

Resumo O objetivo deste estudo foi determinar a presença de ácido desoxirribonucléico (DNA) de Toxoplasma gondii, Sarcocystis spp. e Neospora caninum, em tecidos de javalis abatidos no sul do Brasil. Foram coletadas 156 amostras de diferentes órgãos de 25 javalis, sendo detectado o DNA de pelo menos um dos protozoários pesquisados em 79 amostras. Para diferenciar entre os agentes infecciosos, o polimorfismo do comprimento do fragmento de restrição, foi realizado usando-se as enzimas de restrição DdeI e HpaII. Para N. caninum, a PCR convencional foi realizada com "primers" específicos. O DNA de pelo menos um dos patógenos estudados foi detectado em cada animal: 26,58% para T. gondii, 68,36% para Sarcocystis spp. e 5,06% para N. caninum. Coinfecção entre T. gondii e Sarcocystis spp. ocorreu em 14 animais; entre T. gondii e N. caninum em apenas um animal macho; entre Sarcocystis spp. e N. caninum em uma fêmea, enquanto a coinfecção com os três agentes foi observada igualmente em apenas um animal macho. Considerando-se a alta frequência de detecção e seu risco zoonótico, especialmente T. gondii, constata-se que os javalis podem ser potenciais fontes de transmissão de agentes infecciosos, e a adoção de medidas de monitoramento nessas populações devem ser priorizadas. (DNA spp Brasil 15 2 7 infecciosos usandose usando se HpaII N primers "primers específicos 2658 26 58 26,58 T 6836 68 36 68,36 506 5 06 5,06 1 animais macho fêmea Considerandose Considerando zoonótico constatase constata priorizadas 265 26,5 683 6 3 68,3 50 0 5,0 26, 68, 5,

ABSTRACT Amniotic fluid DNA samples were genotyped by multilocus-nested-PCR-RFLP, but only three of 11 markers amplified 113 of 122 (92.6%) samples, resulting in 12 untyped and 101 partial non-archetypal genotypes. The 101 typed samples were subdivided into four groups: G1 with 73 samples (5’and 3’ SAG2 allele I + SAG3 allele III + GRA6 allele III), 53 had parasite load ≤ 102 parasites/mL (43 asymptomatic, 10 mild infections), 17 had load > 102 and ≤ 103 (one mild, 13 moderate and three severe), and three had load > 103 parasites/mL (three severe); G2 with 22 samples (5’and 3’ SAG2 allele I + SAG3 allele III), all parasite load levels ≤ 102 parasites/mL (18 asymptomatic and four mild); G3 with five samples (5’ and 3’ SAG2 allele I + SAG3 allele II), parasite load ≤ 102 parasites/mL (three asymptomatic and two mild); G4 with one sample (5’ and 3’ SAG2 allele II + SAG3 allele II + GRA6 allele I), a parasite load < 102 parasites/mL in an asymptomatic infant. After DNA sequencing, restriction sites confirmed SAG2, SAG3 and GRA6 alleles in 98.7%, 100% and 100% of the cases, respectively, while single nucleotide polymorphisms confirmed 90% of 5’-SAG2 allele I; 98.7% of 3’-SAG2 allele I; 98% of SAG-3 allele III, but only 40% of GRA6 allele III results. For the moment, partial non-archetypal genotypes of parasites did not show any relationship with either parasite load in amniotic fluid samples or clinical outcome of infants at the age of 12 months. multilocusnestedPCRRFLP, multilocusnestedPCRRFLP multilocus nested PCR RFLP, RFLP multilocus-nested-PCR-RFLP 1 92.6% 926 92 6 (92.6% nonarchetypal non archetypal groups G 7 5and 5 3 SAG GRA , III) parasitesmL mL 43 (4 infections, infections infections) severe, severe severe) 2 18 (1 mild) (5 II, II) I, I) infant sequencing 987 98 100 cases respectively 90 5SAG2 5’-SAG 98.7 3SAG2 3’-SAG SAG- 40 results moment months 92.6 9 (92.6 4 ( 5SAG 98. 3SAG 92. (92. (92 (9