ABSTRACT Objective: Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease and a growing global epidemic. In NAFLD, liver fat surpasses 5% of hepatocytes without the secondary causes of lipid accumulation or excessive alcohol consumption. Given the link between NAFLD and insulin resistance, the possible association between the rs2854744 (−202 G>T) promoter polymorphism of insulin-like growth factor binding protein 3 (IGFBP3) gene and NAFLD was investigated in this study. Materials and methods: In this genetic case-control association study, the IGFBP3 rs2854744 genotypes of 315 unrelated individuals, including 156 patients with biopsy-proven NAFLD and 159 controls, were determined using polymerase chain reaction/restriction fragment length polymorphism analyses. Results: The “GT+TT” genotype of the IGFBP3 rs2854744 polymorphism, compared with the “GG” genotype, was associated with a 2.7-fold increased risk of NAFLD after adjustment for confounding factors (P = 0.009; odds ratio [OR] = 2.71; 95% confidence interval [CI] = 1.19-3.18). Additionally, the IGFBP3 rs2854744 “T” allele, in comparison with the “G” allele, was significantly overrepresented in NAFLD patients than the controls (P = 0.008; OR = 1.85; 95%CI = 1.23-2.94). Conclusion: Our findings first indicated that the IGFBP3 rs2854744 “GT+TT” genotype is a marker of increased NAFLD susceptibility; however, it needs to be supported by further investigations in other populations. Objective (NAFLD epidemic 5 consumption resistance rs rs285474 −202 202 (−20 G>T GT G T insulinlike like IGFBP (IGFBP3 study methods casecontrol case control 31 individuals 15 biopsyproven biopsy proven reactionrestriction reaction restriction analyses Results GT+TT GTTT TT “GT+TT GG “GG 2.7fold 27fold fold 2.7 2 7 P 0.009 0009 0 009 [OR 2.71 271 71 95 CI [CI 1.193.18. 119318 1.19 3.18 . 1 19 18 1.19-3.18) Additionally “T allele “G 0.008 0008 008 1.85 185 85 95CI 1.232.94. 123294 1.23 2.94 23 94 1.23-2.94) Conclusion susceptibility however populations rs28547 −20 20 (−2 (IGFBP 7fold 27 2. 0.00 000 00 9 193 1.193.18 11931 119 1.1 318 3.1 1.19-3.18 1.8 8 232 1.232.94 12329 123 1.2 294 2.9 1.23-2.94 rs2854 −2 (− 0.0 1.193.1 1193 11 1. 3. 1.19-3.1 1.232.9 1232 12 29 1.23-2.9 rs285 − ( 0. 1.193. 1.19-3. 1.232. 1.23-2. rs28 1.193 1.19-3 1.232 1.23-2 rs2 1.19- 1.23-

Abstract The work aims to analyze the associations of polymorphic variants of the PRL and BLG genes with resistance and susceptibility to mastitis in Holstein cows. The experimental study consisted of the selection of biomaterial samples from 250 heads of Holstein cows aged 3 years divided into two groups (healthy and with a confirmed diagnosis of mastitis). The determination of animal genotypes was carried out using polymerase chain reaction and restriction fragment length polymorphism. The study of the nature of the association of polymorphic variants of the PRL and BLG gene with resistance/increased risk of mastitis established a significant deviation from the theoretically expected distribution of bBLG-HaeIII genotypes in the group of animals suffering from mastitis (the value of χ2 was 0.24). The bBLG-HaeIIIBB genotype can act as a marker of an increased risk of developing mastitis in Holstein cows; its frequency in the group of sick animals exceeds the frequency in the control group by more than 2 times (44.0 compared to 17.0%, respectively). The bBLG-HaeIIIAB genotype is significantly associated with mastitis resistance in Holstein cows; its frequency is 2 times lower than in the control group (28.0 compared to 54.0%). 25 healthy mastitis. . mastitis) polymorphism resistanceincreased bBLGHaeIII bBLG HaeIII χ 0.24. 024 0.24 0 24 0.24) bBLGHaeIIIBB HaeIIIBB 44.0 440 44 (44. 170 17 17.0% respectively. respectively respectively) bBLGHaeIIIAB HaeIIIAB 28.0 280 28 (28. 54.0%. 540 54.0% 54 54.0%) 02 0.2 44. 4 (44 1 17.0 28. (28 54.0 5 0. (4 17. (2 54. (

Resumo O trabalho tem como objetivo analisar as associações de variantes polimórficas dos genes PRL e BLG com resistência e suscetibilidade à mastite em vacas holandesas. O estudo experimental consistiu na seleção de amostras de biomateriais de 250 cabeças de vacas holandesas com 3 anos de idade divididas em dois grupos (saudáveis e com diagnóstico confirmado de mastite). A determinação dos genótipos dos animais foi realizada utilizando reação em cadeia da polimerase e polimorfismo de comprimento de fragmentos de restrição. O estudo da natureza da associação de variantes polimórficas do gene PRL e BLG com resistência/risco aumentado de mastite estabeleceu um desvio significativo da distribuição teoricamente esperada dos genótipos bBLG-HaeIII no grupo de animais que sofrem de mastite (o valor de χ2 foi 0,24). O genótipo bBLG-HaeIIIBB pode atuar como marcador de risco aumentado de desenvolvimento de mastite em vacas holandesas; sua frequência no grupo de animais doentes excede a frequência no grupo controle em mais de duas vezes (44,0% contra 17,0%, respectivamente). O genótipo bBLG-HaeIIIAB está significativamente associado à resistência à mastite em vacas holandesas; sua frequência é duas vezes menor que no grupo controle (28,0% contra 54,0%). 25 saudáveis mastite. . mastite) restrição resistênciarisco bBLGHaeIII bBLG HaeIII o χ 0,24. 024 0,24 0 24 0,24) bBLGHaeIIIBB HaeIIIBB 44,0% 440 44 (44,0 170 17 17,0% respectivamente. respectivamente respectivamente) bBLGHaeIIIAB HaeIIIAB 28,0% 280 28 (28,0 54,0%. 540 54,0% 54 54,0%) 2 02 0,2 44,0 4 (44, 1 17,0 28,0 (28, 54,0 5 0, 44, (44 17, 28, (28 54, (4 (2 (

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

Resumo Objetivo: O objetivo deste estudo foi examinar a relação entre os polimorfismos do comprimento do fragmento de restrição (RFLP) BstUI C/T (rs 12722) e RFLP DpnII B1/B2 (rs 13946) COL5A1 e a ruptura do ligamento cruzado anterior (LCA) em atletas de esportes coletivos. Métodos Sessenta e oito atletas de esportes coletivos (n = 36 mulheres e n = 32 homens) com ruptura do LCA (RLCA) sem contato ocorreram durante práticas esportivas (Grupo RLCA) e 42 jogadores saudáveis (n = 20 mulheres e n = 22 homens) (Grupo Controle) participaram do estudo. O DNA genômico foi extraído do swab bucal com o método salting out. Todas as amostras foram genotipadas para os polimorfismos rs12722 e rs13946 por reação em cadeia da polimerase (PCR) e análise de enzimas de restrição. Resultados Nenhuma diferença significativa foi encontrada entre os grupos RLCA e Controle em idade, altura, peso corporal, índice de massa, prática esportiva (horas/semana) e distribuição de gênero entre os diferentes esportes coletivos. O grupo controle teve carreiras esportivas mais longas (p< 0,005). As distribuições de frequência dos polimorfismos de nucleotídeos COL5A1 DpnII estavam em equilíbrio de Hardy-Weinberg (EHW) em ambos os grupos (p do teste de Hardy-Weinberg (HW) > 0,005). As frequências genotípicas de COL5A1 BstUI RFLP C/C foram menores no grupo RLCA em comparação com o grupo Controle (p do teste HW = 0,001). Os genótipos combinados CC, B1B1 mostraram um efeito protetor contra a ruptura do LCA (OR = 83,3 / 16,7 = 5). Conclusões O gene COL5A1 pode ser um dos fatores genéticos associados à RLCA em esportes coletivos. Objetivo (RFLP CT C T rs 12722 B1B2 BB B1 B2 B B1/B 13946 COLA COL A COL5A (LCA 3 homens (RLCA Grupo 4 2 out rs1272 rs1394 PCR (PCR idade altura corporal massa horas/semana horassemana horas semana (horas/semana p< p 0,005. 0005 0,005 . 0 005 0,005) HardyWeinberg Hardy Weinberg EHW (EHW (HW CC 0,001. 0001 0,001 001 0,001) B1B OR 833 83 83, 167 16 7 16, 5. 5 5) 1272 1394 rs127 rs139 000 0,00 00 8 1 127 139 rs12 rs13 0,0 12 13 rs1 0,

Abstract Objective: The aim of this study was to examine the relationship between BstUI restriction fragment length polymorphisms (RFLP) C/T (rs 12722) and DpnII RFLP B1/B2 (rs 13946) COL5A1 polymorphisms and the anterior cruciate ligament (ACL) rupture in competitive team-sport athletes. Methods Sixty-eight team-sport players (n = 36 women and n = 32 men) with non-contact ACL rupture (ACLR) occurred during sport practices (ACLR Group) and 42 healthy players (n = 20 women and n = 22 men) (Control Group) participated in the study. Genomic DNA was extracted from buccal swab with salting out method. All samples were genotyped for the polymorphisms rs12722 and rs13946 by polymerase chain reaction (PCR) and restriction enzymes analysis. Results No significant difference has been found between ACRL and Control groups in age, height, weight body, mass index, sport practice (hours/week) and gender distribution among the different team sports. Control group had longer sport careers (p< 0.005). The frequency distributions of COL5A1 DpnII nucleotide polymorphisms were in Hardy-Weinberg equilibrium (HWE) in both groups (p of the Hardy-Weinberg (HW) -test > 0.005). Genotype frequencies of COL5A1 BstUI RFLP C/C was lower in the ACLR group compared to the Control group (p of the HW-test = 0.001). Combined CC, B1B1 genotypes showed a protective effect against ACL rupture (OR = 83.3 / 16.7 = 5). Conclusions The COL5A1 gene may be one of the genetic factors associated with ACLR in team sport. Objective (RFLP CT C T rs 12722 B1B2 BB B1 B2 B B1/B 13946 COLA COL A COL5A (ACL teamsport athletes Sixtyeight Sixty eight 3 men noncontact non contact Group 4 2 method rs1272 rs1394 PCR (PCR analysis age height body index hours/week hoursweek hours week (hours/week sports p< p 0.005. 0005 0.005 . 0 005 0.005) HardyWeinberg Hardy Weinberg HWE (HWE HW (HW test CC HWtest 0.001. 0001 0.001 001 0.001) B1B OR 833 83 83. 167 16 7 16. 5. 5 5) 1272 1394 rs127 rs139 000 0.00 00 8 1 127 139 rs12 rs13 0.0 12 13 rs1 0.

Abstract Interleukins 6 and 17 act in bone resorption in the presence of infections of endodontic origin for host defense. Genetic polymorphisms may be associated with increased bone loss, represented by areas of large periapical lesions. This study aimed to verify the frequency of interleukin 6 and 17 gene polymorphism in patients with asymptomatic apical periodontitis or chronic apical abscess and to verify the existence of correlations between periapical lesion area with age, gender, and presence of the polymorphism, in the studied population, in the state of Pernambuco. A population consisting of thirty diagnosed individuals was included. The area of the lesions was measured in mm². Genomic DNA was extracted and genotyping was performed by Polymerase Chain Reaction Restriction Fragment Length Polymorphism for interleukin 6 (rs 1800795) and interleukin 17 (rs 2275913). Fisher's exact, chi-square, and odds ratio tests were used. A logistic regression analysis was also performed using sex, age, and the presence of polymorphism as covariates, in addition to linear regression to test the relationship between age and lesion area. All tests used a significance level of 0.05% (p ≤0.05%). There was no statistical significance in the occurrence of large areas of periapical lesions correlated with age, sex, and diagnosis, nor in the distribution of alleles in the polymorphism of interleukins 6 and 17 in the studied groups. The frequency of homozygous and heterozygous polymorphism was high. The polymorphism of these interleukins is not correlated with the increase in the areas of asymptomatic periapical inflammatory lesions. 1 defense loss gender Pernambuco included mm² mm rs 1800795 2275913. 2275913 . 2275913) Fishers Fisher s exact chisquare, chisquare chi square, square chi-square sex covariates 005 0 05 0.05 p ≤0.05%. ≤0.05% ≤0.05%) diagnosis groups high 180079 227591 00 0.0 ≤0.05 18007 22759 0. ≤0.0 1800 2275 ≤0. 180 227 ≤0 18 22 ≤ 2

Resumo As interleucinas 6 e 17 atuam na reabsorção óssea na presença de infecções de oriegem endodôntica para defesa do hospedeiro. Polimorfismos genéticos podem estar associados ao aumento da perda óssea, representada por áreas de lesões periapicais grandes. O objetivo deste estudo foi verificar a frequência do polimorfismo dos genes interleucina 6 e 17 em pacientes com periodontite apical assintomática ou abscesso apical crônico e verificar a existência de correlações entre área de lesão periapical com idade, sexo e presença do polimorfismo, na população estudada, no estado de Pernambuco. Foi incluída uma população constituída por trinta indivíduos diagnosticados. A áreas da lesões foram medidas em mm². O DNA genômico foi extraído e a genotipagem realizada por Polimorfismo de Comprimento de Fragmento de Restrição de Reação em Cadeia da Polimerase para interleucina 6 (rs 1800795) e interleucina 17 (rs 2275913). Os testes exato de Fisher, qui-quadrado e odds ratio foram utilizados. Uma análise de regressão logística também foi realizada usando sexo, idade e presença de polimorfismo como covariável, além de regressão linear para testar a relação da idade e área da lesão. Todos os testes utilizaram um nível de significância de 0,05% (p ≤0.05%). Não houve significância estatística na ocorrência das áreas grandes de lesões periapicais correlacionadas com idade, sexo e diagnóstico nem nas distribuições de alelos no polimorfismo das interleucinas 6 e 17 nos grupos estudados. A frequência de polimorfismo homozigoto e heterozigoto foi alta. O polimorfismo dessas interleucinas não está correlacionado ao aumento das áreas das lesões inflamatórias periapicais assintomáticas. 1 hospedeiro estudada Pernambuco diagnosticados mm² mm rs 1800795 2275913. 2275913 . 2275913) Fisher quiquadrado qui quadrado utilizados covariável 005 0 05 0,05 p ≤0.05%. ≤0.05% ≤0.05%) estudados alta assintomáticas 180079 227591 00 0,0 ≤0.05 18007 22759 0, ≤0.0 1800 2275 ≤0. 180 227 ≤0 18 22 ≤ 2

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.

SUMMARY OBJECTIVE: The aim of this study was to determine frequency and associations between APOA5 c.56C>G, −1131T>C, c.553G>T, and APOC3 −482C>T and SstI gene polymorphisms with hypertriglyceridemia. METHODS: Under a case-control study model, 135 hypertriglyceridemic and 178 normotriglyceridemic control participants were recruited. Polymerase chain reaction and restriction fragment length polymorphism methods were utilized for genotyping. Statistical calculations were performed by comparing allele and genotype frequencies between groups. Clinical characteristics were compared between groups and intra-group genotypes. RESULTS: APOC3 gene −482C>T and SstI polymorphic genotypes and allele frequencies were significantly higher in hypertriglyceridemic group (genotype frequencies, p=0.035, p=0.028, respectively). Regression analysis under unadjusted model confirmed that APOC3 −482C>T and SstI polymorphisms were significantly contributing to have hypertriglyceridemia (p=0.02, odds ratio [OR]=1.831 (95% confidence interval [CI] 1.095–3.060); p=0.04, OR=1.812 (1.031–3.183), respectively). APOA5 c.56C>G was in complete linkage disequilibrium with APOA5 c.553G>T polymorphism (D’=1). CONCLUSION: For the first time in a population sample from Turkey, among the five polymorphisms of APOA5 and APOC3 genes investigated, APOC3 −482C>T and SstI polymorphisms were associated with elevated serum TG levels, while APOA5 c.56C>G, −1131T>C, and c.553G>T polymorphisms were not. OBJECTIVE APOA c56CG cCG c 56C G C 1131TC TC 1131T T −1131T>C c553GT cGT 553G APOC 482CT CT 482C METHODS casecontrol case 13 17 recruited genotyping intragroup intra RESULTS p0035 p 0 035 p=0.035 p0028 028 p=0.028 respectively. respectively . respectively) p=0.02, p002 02 (p=0.02 OR=1.831 OR1831 OR =1.831 1 831 [OR]=1.83 95% 95 (95 CI [CI 1.095–3.060 10953060 095 3 060 1.095–3.060) p004 04 p=0.04 OR1812 812 OR=1.81 1.031–3.183, 10313183 1.031–3.183 , 031 183 (1.031–3.183) D=1. D1 D =1 (D’=1) CONCLUSION Turkey investigated levels not CG GT p003 03 p=0.03 p=0.02 p00 (p=0.0 OR=1.83 OR183 1831 =1.83 83 [OR]=1.8 9 (9 1.095–3.06 1095306 09 06 p=0.0 OR181 81 OR=1.8 1031318 1.031–3.18 18 (1.031–3.183 D=1 = (D’=1 p0 (p=0. OR18 =1.8 8 [OR]=1. ( 1.095–3.0 109530 p=0. OR=1. 103131 1.031–3.1 (1.031–3.18 D= (D’= (p=0 OR1 =1. [OR]=1 1.095–3. 10953 p=0 OR=1 10313 1.031–3. (1.031–3.1 (D’ (p= [OR]= 1.095–3 1095 p= OR= 1031 1.031–3 (1.031–3. (D (p [OR] 1.095– 109 103 1.031– (1.031–3 [OR 1.095 10 1.031 (1.031– 1.09 1.03 (1.031 1.0 (1.03 1. (1.0 (1. (1

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 Background The p53 and p21 proteins are important regulators of cell cycle and apoptosis and may contribute to autoimmune diseases, such as systemic lupus erythematosus (SLE). As genetic polymorphisms may cause changes in protein levels and functions, we investigated associations of TP53 and p21 (CDKN1A) polymorphisms (p53 72 G > C—rs1042522; p53 PIN3—rs17878362; p21 31 C > A—rs1801270; p21 70 C > T—rs1059234) with the development of systemic lupus erythematosus (SLE) in a Southeastern Brazilian population. Methods Genotyping of 353 female volunteers (cases, n = 145; controls, n = 208) was performed by polymerase chain reaction, restriction fragment length polymorphism and/or DNA sequencing. Associations between TP53 and p21 polymorphisms and SLE susceptibility and clinical manifestations of SLE patients were assessed by logistic regression analysis. Results Protective effect was observed for the genotype combinations p53 PIN3 A1/A1 -p21 31 C/A, in the total study population (OR 0.45), and p53 PIN3 A1/A2-p21 31 C/C, in non-white women (OR 0.28). In Whites, p53 72 C-containing (OR 3.06) and p53 PIN3 A2-containing (OR 6.93) genotypes were associated with SLE risk, and higher OR value was observed for the combined genotype p53 72 G/C-p53 PIN3 A1/A2 (OR 9.00). Further, p53 PIN3 A1/A2 genotype was associated with serositis (OR 2.82), while p53 PIN3 A2/A2 and p53 72 C/C genotypes were associated with neurological disorders (OR 4.69 and OR 3.34, respectively). Conclusions Our findings showed that the TP53 and p21 polymorphisms included in this study may have potential to emerge as SLE susceptibility markers for specific groups of patients. Significant interactions of the TP53 polymorphisms with serositis and neurological disorders were also observed in SLE patients. Highlights The polymorphisms TP53 rs1042522 (G > C) and TP53 rs17878362 (16 bp Del/Ins) were associated with SLE risk in whites. In whites, the combined genotype TP53 rs1042522 GC- TP53 rs17878362 A1A2 and the haplotype TP53 rs1042522 C-rs17878362 A2 represented higher SLE risk. Combination of TP53 rs17878362 (16 bp Del/Ins) and p21 rs1801270 (C > A) protected against SLE in non-white women. TP53 and p21 (CDKN1A) polymorphisms may be SLE susceptibility markers for specific groups. p p5 p2 diseases SLE. . functions TP TP5 CDKN1A CDKNA CDKN A (CDKN1A (p5 7 Crs1042522 Crs rs C—rs1042522 PIN3rs17878362 PINrs PIN PIN3—rs17878362 3 Ars1801270 Ars A—rs1801270 Trs1059234 Trs T rs1059234 T—rs1059234 (SLE 35 cases, cases (cases 145 controls 208 reaction andor or sequencing analysis A1A1 AA A1 A1/A -p2 CA C/A 0.45, 045 0.45 , 0 45 0.45) A1/A2p21 A1A2p21 AAp A1/A2-p2 CC nonwhite non white 0.28. 028 0.28 28 0.28) Whites Ccontaining containing 3.06 306 06 A2containing Acontaining 6.93 693 6 93 G/Cp53 GCp53 GCp G/C G/C-p5 9.00. 900 9.00 9 00 9.00) Further 2.82, 282 2.82 2 82 2.82) A2A2 A2/A 469 4 69 4.6 334 34 3.34 respectively. respectively respectively) rs104252 rs1787836 16 (1 Del/Ins DelIns Del Ins whites GC A1A Crs17878362 C-rs1787836 rs180127 (p Crs104252 C—rs104252 PIN3rs1787836 PIN3—rs1787836 Ars180127 A—rs180127 Trs105923 rs105923 T—rs105923 14 20 -p 04 0.4 A2p21 A1/A2p2 A1A2p2 A1/A2-p 02 0.2 3.0 30 6.9 Cp53 Cp G/Cp5 GCp5 G/C-p 90 9.0 2.8 8 A2A 46 4. 33 3.3 rs10425 rs178783 1 ( Crs1787836 C-rs178783 rs18012 Crs10425 C—rs10425 PIN3rs178783 PIN3—rs178783 Ars18012 A—rs18012 Trs10592 rs10592 T—rs10592 0. Ap A2p2 A1/A2p A1A2p 3. 6. Cp5 G/Cp 9. 2. rs1042 rs17878 Crs178783 C-rs17878 rs1801 Crs1042 C—rs1042 PIN3rs17878 PIN3—rs17878 Ars1801 A—rs1801 Trs1059 rs1059 T—rs1059 A2p rs104 rs1787 Crs17878 C-rs1787 rs180 Crs104 C—rs104 PIN3rs1787 PIN3—rs1787 Ars180 A—rs180 Trs105 rs105 T—rs105 rs10 rs178 Crs1787 C-rs178 rs18 Crs10 C—rs10 PIN3rs178 PIN3—rs178 Ars18 A—rs18 Trs10 T—rs10 rs1 rs17 Crs178 C-rs17 Crs1 C—rs1 PIN3rs17 PIN3—rs17 Ars1 A—rs1 Trs1 T—rs1 Crs17 C-rs1 C—rs PIN3rs1 PIN3—rs1 A—rs T—rs C-rs PIN3rs PIN3—rs

Abstract Milk’s qualitative and technological properties are greatly affected by genetic polymorphisms in the kappa-casein gene, and their polymorphisms may serve as informative markers of yield and composition. Thus, the objective of this study was to detect kappa-casein (kappa-CN) gene polymorphisms and their association with milk production traits in crossbred dairy cows. One hundred healthy crossbred (Friesian x Jenoubi) dairy animals between three and five years old were sampled for blood and milk during their first lactation. The genomic DNA was extracted from whole blood, and restriction fragment length polymorphism (RFLP-PCR) was used to determine the genotype of the kappa-CN gene. As a consequence of the restriction digestion of this fragment with Hind III, it showed three different restriction patterns: BB (453 base pairs uncut), AB (453, 206, and 225 base pairs), and AA (206 and 225 base pairs). Based on genetic diversity, the AB genotype was the most predominant (n = 67), with a frequency of 0.67. A variant genotype of the kappa-CN gene was associated with milk production traits in crossbred dairy cows. Animals with the AA variant produced a higher milk yield and a higher percentage of fat, casein, protein, and solids not fat (SNF) (P≤0.05) (1.397kg, 0.75%, 0.31%, 0.27%, and 0.68%, respectively) than those with the BB variant. A logistic regression analysis confirmed that the kappa-CN genotypes increase milk yield and casein content. Therefore, genetic variants of the kappa-CN gene could be used as genetic markers for improving milk production traits in dairy cattle. Milks Milk s kappacasein kappa composition Thus kappaCN CN (kappa-CN cows Friesian Jenoubi lactation RFLPPCR RFLP PCR (RFLP-PCR III patterns 453 (45 uncut, uncut , uncut) 453, 206 22 pairs, pairs) (20 pairs. . diversity n 67, 67 67) 067 0 0.67 protein SNF (SNF P≤0.05 P005 P 05 (P≤0.05 1.397kg, 1397kg kg 1 397kg (1.397kg 075 75 0.75% 031 31 0.31% 027 27 0.27% 068 68 0.68% respectively content Therefore cattle 45 (4 20 2 (2 6 06 0.6 P≤0.0 P00 (P≤0.0 1.397kg 07 7 0.75 03 3 0.31 02 0.27 0.68 4 ( 0. P≤0. P0 (P≤0. 0.7 0.3 0.2 P≤0 (P≤0 P≤ (P≤ (P

Resumo As propriedades qualitativas e tecnológicas do leite são muito afetadas por polimorfismos genéticos no gene kappa-caseína e esses polimorfismos podem servir como marcadores informativos de rendimento e composição. Assim, o objetivo deste estudo foi detectar polimorfismos do gene kappa-caseína (kappa-CN) e sua associação com características de produção de leite em vacas leiteiras mestiças. Cem animais mestiços (Freisian x Jenoubi) saudáveis, entre três e cinco anos de idade, foram amostrados durante a primeira lactação para sangue e leite. O DNA genômico foi extraído do sangue total e o polimorfismo dos fragmentos de restrição do DNA genômico (RFLP-PCR) foi usado para determinar o genótipo do gene kappa-CN. Em consequência da digestão de restrição deste fragmento com Hind III, ele apresentou três padrões de restrição diferentes: BB (453 pares de bases não cortadas), AB (453,206 e 225 pares de bases) eAA (206 e 225 pares de bases). Com base na diversidade genética, o genótipo AB foi o mais predominante (n = 67), com frequência de 0,67. Genótipo variante do gene kappa-CN foi associado com características de produção de leite em vacas leiteiras mestiças. Animais com a variante AA tiveram maior produção de leite e maior percentual de gordura, caseína, proteína e sólidos não gordurosos (SNG) (P≤0,05) (l,397kg, 0,75%, 0,31%, 0,27% e 0,68%, respectivamente) do que aqueles com variante BB. Uma análise de regressão logística confirmou que os genótipos kappa-CN aumentam a produção de leite e o teor de caseína. Portanto, variantes genéticas do gene kappa-CN podem ser usadas como marcadores genéticos para melhorar as características de produção de leite em bovinos leiteiros. kappacaseína kappa caseína composição Assim kappaCN CN (kappa-CN mestiças Freisian Jenoubi saudáveis idade RFLPPCR RFLP PCR (RFLP-PCR kappaCN. CN. III diferentes 453 (45 cortadas, cortadas , cortadas) 453,206 453206 206 (453,20 22 (20 bases. . genética n 67, 67 67) 067 0 0,67 gordura SNG (SNG P≤0,05 P005 P 05 (P≤0,05 l,397kg, l397kg lkg l 397kg kg (l,397kg 075 75 0,75% 031 31 0,31% 027 27 0,27 068 68 0,68% respectivamente Portanto leiteiros 45 (4 453,20 45320 20 (453,2 2 (2 6 06 0,6 P≤0,0 P00 (P≤0,0 l,397kg 07 7 0,75 03 3 0,31 02 0,2 0,68 4 ( 453,2 4532 (453, 0, P≤0, P0 (P≤0, 0,7 0,3 453, P≤0 (P≤0 P≤ (P≤ (P

SUMMARY OBJECTIVE: Recurrent pregnancy loss is considerably a reproductive health problem for couples. Genetic, epigenetic, and environmental factors play an important role in the development of recurrent pregnancy loss. While there are many causes, genetic and epigenetic factors are common. In this study, we aimed to examine the association between miR604 (rs2368393) A>G gene polymorphism and the risk of recurrent miscarriage in the Turkish population. METHODS: The study included 250 participants (i.e., 150 patients and 100 controls). DNA samples were isolated from peripheral blood, and polymerase chain reactions and restriction fragment length polymorphism methodologies were applied. RESULTS: The genotype distribution and allele frequencies of miR604A>G gene showed statistically significant differences between patients and control groups (p=0.002 and p<0.002, respectively). CONCLUSION: As a result of the study, we found that the AA genotype and A allele of the miR604A>G gene were statistically significant for the risk of recurrent pregnancy loss in Turkish women. OBJECTIVE couples Genetic causes common miR miR60 rs2368393 rs (rs2368393 AG G population METHODS 25 i.e., ie i e (i.e. 15 10 controls. controls . controls) blood applied RESULTS miR604AG miRAG miR604A p=0.002 p0002 p 0 002 (p=0.00 p<0.002 respectively. respectively respectively) CONCLUSION women miR6 rs236839 (rs236839 2 i.e. (i.e 1 miRA p=0.00 p000 00 (p=0.0 p<0.00 rs23683 (rs23683 i.e p=0.0 p00 (p=0. p<0.0 rs2368 (rs2368 p=0. p0 (p=0 p<0. rs236 (rs236 p=0 (p= p<0 rs23 (rs23 p= (p p< rs2 (rs2 (rs

Abstract The aim of this study was to identify Cryptosporidium species found in cattle and sheep in Paraná, southern region of Brazil. Individual fecal samples from 458 bovines and 101 sheep were submitted for molecular analysis by PCR and nested PCR using specific primers for sequences of the 18S ribosomal unit (rRNA). Positive samples were analyzed using restriction fragment length polymorphism (RFLP), followed by genetic sequencing for species confirmation. The occurrence of Cryptosporidium was 11.27% (63/559). The highest occurrence was detected in lambs (12/59, 20.33%). From the 63 positive samples, it was possible to identify the species in 58 of them by RFLP and genetic sequencing. Five species of Cryptosporidium were identified: Cryptosporidium andersoni, Cryptosporidium bovis, Cryptosporidium ryanae, Cryptosporidium xiaoi, and Cryptosporidium parvum. The most prevalent species was C. andersoni (41.38%) and the least predominant was C. parvum (10.34%). The most abundant species of Cryptosporidium in dairy calves were C. andersoni (11/25) and C. ryanae (6/25). Of the 17 positive sheep, nine (52.94%) were infected with C. andersoni. This finding is the first report on the occurrence of C. andersoni in naturally infected sheep in Brazil and the first observation of a high absolute occurrence of this Cryptosporidium species in sheep. Paraná 45 10 S rRNA. rRNA . (rRNA) RFLP, , (RFLP) confirmation 1127 11 27 11.27 63/559. 63559 63/559 559 (63/559) 12/59, 1259 12 59 (12/59 20.33%. 2033 20.33% 20 33 20.33%) 6 5 identified bovis xiaoi C 41.38% 4138 41 38 (41.38% 10.34%. 1034 10.34% 34 (10.34%) 11/25 1125 25 (11/25 6/25. 625 6/25 (6/25) 1 52.94% 5294 52 94 (52.94% 4 (rRNA (RFLP 112 2 11.2 6355 63/55 55 (63/559 12/59 125 (12/5 203 20.33 3 41.38 413 (41.38 103 10.34 (10.34% 11/2 (11/2 62 6/2 (6/25 52.94 529 9 (52.94 11. 635 63/5 (63/55 12/5 (12/ 20.3 41.3 (41.3 10.3 (10.34 11/ (11/ 6/ (6/2 52.9 (52.9 63/ (63/5 12/ (12 20. 41. (41. 10. (10.3 (11 (6/ 52. (52. (63/ (1 (41 (10. (6 (52 (63 ( (4 (10 (5

Resumo O objetivo deste estudo foi identificar espécies de Cryptosporidium em bovinos e ovinos do Paraná, região sul do Brasil. Amostras de fezes de 458 bovinos e 101 ovinos foram individualmente submetidas à análise molecular por PCR e nested PCR, utilizando-se iniciadores específicos para sequências da unidade ribossomal 18S (rRNA). As amostras positivas foram analisadas pelo polimorfismo de comprimento de fragmento de restrição (RFLP), seguido de sequenciamento genético para confirmação da espécie. A ocorrência de Cryptosporidium foi de 11,27% (63/559). Observou-se maior ocorrência em cordeiros (20,33%). Das 63 amostras positivas, foi possível identificar as espécies em 58 amostras por RFLP e sequenciamento genético. Foram identificadas cinco espécies de Cryptosporidium: Cryptosporidium andersoni, Cryptosporidium bovis, Cryptosporidium ryanae, Cryptosporidium xiaoi e Cryptosporidium parvum. A espécie mais predominantemente encontrada foi C. andersoni (41,38%) e a menos foi C. parvum (10,34%). As espécies mais abundantes de Cryptosporidium, em bezerros leiteiros, foram C. andersoni (11/25) e C. ryanae (6/25). Dos 17 ovinos positivos, nove (52,94%) estavam infectados com C. andersoni. Este achado é o primeiro relato sobre a ocorrência de C. andersoni em ovinos naturalmente infectados no Brasil e a primeira observação de alta ocorrência absoluta desta espécie de Cryptosporidium em ovinos. Paraná 45 10 utilizandose utilizando se S rRNA. rRNA . (rRNA) RFLP, , (RFLP) 1127 11 27 11,27 63/559. 63559 63/559 559 (63/559) Observouse Observou 20,33%. 2033 20,33% 20 33 (20,33%) 6 5 bovis C 41,38% 4138 41 38 (41,38% 10,34%. 1034 10,34% 34 (10,34%) leiteiros 11/25 1125 25 (11/25 6/25. 625 6/25 (6/25) 1 positivos 52,94% 5294 52 94 (52,94% 4 (rRNA (RFLP 112 2 11,2 6355 63/55 55 (63/559 203 20,33 3 (20,33% 41,38 413 (41,38 103 10,34 (10,34% 11/2 (11/2 62 6/2 (6/25 52,94 529 9 (52,94 11, 635 63/5 (63/55 20,3 (20,33 41,3 (41,3 10,3 (10,34 11/ (11/ 6/ (6/2 52,9 (52,9 63/ (63/5 20, (20,3 41, (41, 10, (10,3 (11 (6/ 52, (52, (63/ (20, (41 (10, (1 (6 (52 (63 (20 (4 (10 ( (5 (2

Abstract The quality and quantity of the extracted DNA are two key aspects for a successful PCR (Polymerase Chain Reaction) amplification. Moreover, reduction in time and cost required for DNA extraction are also two considerable factors, in cases when large number of samples are to be analyzed within a limited time-frame and budget. Accordingly, the aim of this study was to compare and optimize performance of five different DNA extraction methods by boiling meat tissues from cattle, buffalo, sheep, goat, chicken, camel, horse and dog in PBS (Phosphate Buffer Saline), distilled water, alkaline lysis buffers 1, 2 or 3. The results indicated that the boiling of meat and its products in alkaline lysis buffers was a good method to extract crude DNA. The optimized crude DNA extraction protocol was coupled with PCR-RFLP (Restriction Fragment Length Polymorphism) analysis for meat species differentiation. This developed workflow was tested on fifty-three commercial beef and mutton samples, out of which three samples were found to be adulterated. In conclusion, the rapid crude DNA extraction protocol has led to the development of a direct PCR-RFLP workflow that is simple, time-saving and cost-effective for PCR-based identification of different meat species. Polymerase Reaction amplification Moreover factors timeframe frame budget Accordingly cattle buffalo sheep goat chicken camel Phosphate Saline, Saline , Saline) water 1 3 PCRRFLP RFLP Restriction Polymorphism differentiation fiftythree fifty adulterated conclusion simple timesaving saving costeffective effective PCRbased based