ABSTRACT Objective: To investigate the association of the rs741301 polymorphism in the ELMO1 gene with diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM). Materials and methods: This study analyzed 350 patients with T2DM and DR (cases) and 234 patients with T2DM without this complication but with more than 10 years of diabetes mellitus (DM) (controls). DR was diagnosed by indirect fundoscopy. Genotyping was performed by allelic discrimination real-time PCR. Results: The frequency of the C/C genotype of the rs741301 polymorphism in the ELMO1 gene was 26.9% in cases and 17.9% in controls (P = 0.011). After adjustment for covariables, the C/C genotype was associated with an increased risk of DR [odds ratio (OR) = 1.805, 95%CI 1.101–2.961; P = 0.019]. This association remained significant in dominant and additive inheritance models after adjustment for the same variables [OR = 1.597, 95%CI 1.089-2.343; P = 0.017; and OR = 1.818, 95%CI 1.099-3.007; P = 0.020]. Conclusion: This study demonstrated an association between the presence of the C allele of the ELMO1 rs741301 polymorphism and an increased risk of DR in patients with T2DM from Southern Brazil. Objective rs rs74130 ELMO (DR T2DM. TDM . T DM (T2DM) methods 35 (cases 23 1 (DM controls. (controls) fundoscopy realtime real time PCR Results CC 269 26 9 26.9 179 17 17.9 0.011. 0011 0.011 0 011 0.011) covariables odds (OR 1805 805 1.805 95CI CI 95 1.101–2.961 11012961 101 961 0.019. 0019 0.019 019 0.019] 1597 597 1.597 1.0892.343 10892343 1.089 2.343 089 343 1.089-2.343 0.017 0017 017 1818 818 1.818 1.0993.007 10993007 1.099 3.007 099 3 007 1.099-3.007 0.020. 0020 0.020 020 0.020] Conclusion Brazil rs7413 (T2DM (controls 26. 17. 001 0.01 01 180 80 1.80 1.101–2.96 1101296 96 159 59 1.59 0892 1.0892.34 1089234 1089 1.08 2343 2.34 08 34 1.089-2.34 181 81 1.81 0993 1.0993.00 1099300 1099 1.09 3007 3.00 09 00 1.099-3.00 002 0.02 02 rs741 0.0 18 8 1.8 1.101–2.9 110129 15 5 1.5 1.0892.3 108923 108 1.0 2.3 1.089-2.3 1.0993.0 109930 109 300 3.0 1.099-3.0 rs74 0. 1. 1.101–2. 11012 1.0892. 10892 2. 1.089-2. 1.0993. 10993 30 3. 1.099-3. rs7 1.101–2 1101 1.0892 1.089-2 1.0993 1.099-3 1.101– 110 1.089- 1.099- 1.101 11 1.10 1.1

ABSTRACT Objective: To investigate the association between the long noncoding RNAs (lncRNAs) maternally expressed gene 3 (MEG3) rs7158663 polymorphism and diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM). Subjects and methods: The study included 628 patients with T2DM and DR ("case group," including 283 with proliferative DR [PDR] and 345 with nonproliferative DR [NPDR]), and 381 patients with T2DM but no DR ("control group"). The diagnosis of DR was established using indirect ophthalmoscopy. The rs7158663 A/G polymorphism was genotyped using real-time polymerase chain reaction (PCR) with TaqMan probes. Results: Patients with DR, compared with those without DR, had lower frequencies of both the G/G genotype (17.5% and 23.6%, respectively, p = 0.044) and the G allele (p = 0.017). When only patients with PDR were compared with controls, the G/G genotype was associated with increased protection against PDR after adjustment (odds ratio 0.551, 95% confidence interval 0.314–0.966, p = 0.038). This association also remained in the dominant (p = 0.036) and additive (p = 0.031) genetic models. Conclusion: This study reveals, for the first time, that the G/G genotype of the lncRNA MEG3 rs7158663 single-nucleotide polymorphism is associated with a protective effect against advanced-stage DR in patients with T2DM. Additional studies are warranted to validate this finding. Objective lncRNAs (lncRNAs MEG (MEG3 rs rs715866 (DR TDM . T DM (T2DM) methods 62 case group, group 28 [PDR 34 NPDR, NPDR , [NPDR]) 38 control group. group") ophthalmoscopy AG A realtime real time PCR (PCR probes Results GG 17.5% 175 17 5 (17.5 236 23 6 23.6% respectively 0.044 0044 0 044 0.017. 0017 0.017 017 0.017) controls odds 0551 551 0.551 95 03140966 314 966 0.314–0.966 0.038. 0038 0.038 038 0.038) 0.036 0036 036 0.031 0031 031 models Conclusion reveals singlenucleotide single nucleotide advancedstage advanced stage finding (MEG rs71586 (T2DM [NPDR] group" 17.5 1 (17. 23.6 0.04 004 04 001 0.01 01 055 55 0.55 9 0314096 31 96 0.314–0.96 003 0.03 03 rs7158 [NPDR 17. (17 23. 0.0 00 05 0.5 031409 0.314–0.9 rs715 (1 0. 03140 0.314–0. rs71 ( 0314 0.314–0 rs7 0.314– 0.314 0.31 0.3

Abstract Long non-coding RNAs (lncRNAs) are RNAs with >200 nucleotides that are unable to encode proteins and are involved in gene expression regulation. LncRNAs have a key role in many physiological and pathological processes and, consequently, they have been associated with several human diseases, including diabetes chronic complications, such as diabetes kidney disease (DKD). In this context, some studies have identified the dysregulation of the lncRNAs MALAT1 and TUG1 in patients with DKD; nevertheless, available data are still contradictory. Thus, the objective of this study was to compare MALAT1 and TUG1 expressions in urine of patients with type 1 diabetes mellitus (T1DM) categorized according to DKD presence. This study comprised 18 T1DM patients with DKD (cases) and 9 long-duration T1DM patients without DKD (controls). MALAT1 and TUG1 were analyzed using qPCR. Bioinformatics analyses were done to identify both lncRNA target genes and the signaling pathways under their regulation. The lncRNA MALAT1 was upregulated in urine of T1DM patients with DKD vs. T1DM controls (P = 0.007). The expression of lncRNA TUG1 did not differ between groups (P = 0.815). Bioinformatics analysis showed these two lncRNAs take part in metabolism-related pathways. The present study shows that the lncRNA MALAT1 is upregulated in T1DM patients presenting DKD. noncoding non coding (lncRNAs 200 >20 regulation consequently diseases complications . (DKD) context MALAT TUG nevertheless contradictory Thus TDM T DM (T1DM presence cases (cases longduration long duration controls. (controls) qPCR vs P 0.007. 0007 0.007 0 007 0.007) 0.815. 0815 0.815 815 0.815) metabolismrelated metabolism related 20 >2 (DKD (controls 000 0.00 00 081 0.81 81 2 > 0.0 08 0.8 8 0.

ABSTRACT Objective: The objective of this study was to investigate the association between SNPs in the TIE2 and ANGPT-1 genes and diabetic retinopathy (DR). Subjects and methods: This study comprised 603 patients with type 2 diabetes mellitus (T2DM) and DR (cases) and 388 patients with T2DM for more than 10 years and without DR (controls). The TIE2 rs639225 (A/G) and rs638203 (A/G) SNPs and the ANGPT-1 rs4324901 (G/T) and rs2507800 (T/A) SNPs were genotyped by real-time PCR using TaqMan MGB probes. Results: The G/G genotype of the rs639225/TIE2, the G/G genotype of the rs638203/TIE2 and the T allele of the rs4324901/ANGPT-1 SNPs were associated with protection against DR after adjustment for age, glycated hemoglobin, gender, and presence of hypertension (P = 0.042, P = 0.003, and P = 0.028, respectively). No association was found between the rs2507800/ANGPT-1 SNP and DR. Conclusion: We demonstrated, for the first time, the association of TIE2 rs638203 and rsrs939225 SNPs and ANGPT-1 rs4324901 SNP with protection against DR in a Brazilian population. Objective TIE ANGPT1 ANGPT 1 ANGPT- . (DR) methods 60 TDM DM (T2DM cases (cases 38 controls. controls (controls) rs rs63922 A/G AG A G (A/G rs63820 rs432490 G/T GT (G/T rs250780 T/A TA (T/A realtime real time probes Results GG rs639225TIE2 rsTIE rs639225/TIE2 rs638203TIE2 rs638203/TIE rs4324901/ANGPT1 rs4324901ANGPT1 rsANGPT rs4324901/ANGPT rs4324901/ANGPT- age hemoglobin gender 0042 0 042 0.042 0003 003 0.003 0028 028 0.028 respectively. respectively respectively) rs2507800/ANGPT1 rs2507800ANGPT1 rs2507800/ANGPT rs2507800/ANGPT- Conclusion demonstrated rsrs rsrs93922 population (DR 6 3 (controls rs6392 rs6382 rs43249 rs25078 rs639225TIE rs639225/TIE rs638203TIE rs4324901ANGPT 004 04 0.04 000 00 0.00 002 02 0.02 rs2507800ANGPT rsrs9392 rs639 rs638 rs4324 rs2507 0.0 rsrs939 rs63 rs432 rs250 0. rsrs93 rs6 rs43 rs25 rsrs9 rs4 rs2

Abstract The transforming growth factor beta 1 (TGFB1) is a pro-inflammatory cytokine that plays a key role in the mechanisms of angiogenesis and breakdown of the blood-retina barrier, which are implicated in the pathogenesis of diabetic retinopathy (DR). Polymorphisms in the TGFB1 gene have been associated with DR; however, results are still contradictory. Therefore, the aim of this study was to investigate the potential association between two TGFB1 polymorphisms and DR. This study included 992 patients with diabetes mellitus (DM): 546 patients with DR (cases) and 446 patients without DR and with ≥10 years of DM (controls). The TGFB1 rs1800469 and rs1800470 polymorphisms were genotyped by real-time PCR. Frequency of rs1800469 T/T genotype was higher in controls compared to DR cases (18.3% vs. 12.7%, P= 0.022). This genotype remained associated with protection for DR, adjusting for covariables (OR= 0.604; 95% CI 0.395 - 0.923; P= 0.020, recessive model). The rs1800470 C/C genotype was observed in 25.4% of the controls and 18.0% of the cases (P= 0.015); thus, being associated with protection against DR under the recessive model (OR= 0.589; 95% CI 0.405 - 0.857; P= 0.006), adjusting for covariables. In conclusion, the TGFB1 rs1800469 and rs1800470 polymorphisms are associated with protection against DR in DM patients from Southern Brazil. TGFB (TGFB1 proinflammatory pro inflammatory bloodretina blood retina barrier . (DR) however contradictory Therefore 99 (DM) 54 (cases 44 10 ≥1 controls. (controls) rs rs180046 rs180047 realtime real time PCR TT T 18.3% 183 18 3 (18.3 vs 127 12 7 12.7% P 0.022. 0022 0.022 0 022 0.022) OR= OR (OR 0.604 0604 604 95 0395 395 0.39 0.923 0923 923 0020 020 0.020 model. model) CC C 254 25 4 25.4 180 18.0 (P 0.015 0015 015 0.015) thus 0.589 0589 589 0405 405 0.40 0.857 0857 857 0.006, 0006 0.006 , 006 0.006) conclusion Brazil (TGFB (DR 9 (DM 5 ≥ (controls rs18004 18.3 (18. 12.7 002 0.02 02 0.60 060 60 039 39 0.3 0.92 092 92 2 25. 18. 0.01 001 01 0.58 058 58 040 40 0.4 0.85 085 85 000 0.00 00 rs1800 (18 12. 0.0 0.6 06 6 03 0. 0.9 09 0.5 05 04 0.8 08 8 rs180 (1 rs18 ( rs1

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.

ABSTRACT Objective: As studies have reported the involvement of angiopoietin-2 (ANGPT-2) in the pathogenesis of diabetic retinopathy (DR), the aim of this study was to investigate the association between the ANGPT-2 rs2442598 polymorphism and DR. Materials and methods: This case-control study comprised 107 patients with type 1 diabetes mellitus (T1DM) and DR (cases) and 129 patients with T1DM without DR (controls) and with ≥ 10 years of DM. The ANGPT-2 rs2442598 (G/A) polymorphism was genotyped by real-time PCR using TaqMan MGB probes. Results: Genotype distributions of this polymorphism were consistent with the Hardy-Weinberg equilibrium. The frequency of the rs2442598 A allele was higher in cases compared to controls (p = 0.011). Moreover, the A/A genotype was more frequent in cases than in controls (p = 0.017) and was associated with risk for DR after adjustments for duration of DM, HbA1c, triglycerides, estimated glomerular filtration rate, and hypertension (odds ratio [OR] = 5.19, 95% confidence interval [CI] 1.21-22.27). This association was maintained under recessive (OR = 4.78, 95% CI 1.14-19.99) and additive (OR = 6.861, 95% CI 1.45-32.38) inheritance models. Conclusion: Our data demonstrated, for the first time, an association between the ANGPT-2 rs2442598 A allele and risk for DR in T1DM patients from southern Brazil. Additional studies are necessary to replicate this association in other populations.

Abstract Autoimmune diseases are characterized by the loss of self-tolerance, leading to immune-mediated tissue destruction and chronic inflammation. Tyrosine kinase 2 (TYK2) protein plays a key role in immunity and apoptosis pathways. Studies have reported associations between single nucleotide polymorphisms (SNPs) in the TYK2 gene and autoimmune diseases; however, results are still inconclusive. Thus, we conducted a systematic review followed by meta-analysis. A literature search was performed to find studies that investigated associations between TYK2 SNPs and autoimmune diseases (multiple sclerosis, systemic lupus erythematosus, Crohn’s disease, ulcerative colitis, psoriasis, rheumatoid arthritis, type 1 diabetes, and inflammatory bowel disease). Pooled odds ratios (OR) with 95 % CI were calculated using random (REM) or fixed (FEM) effects models in the Stata 11.0 Software. Thirty-four articles were eligible for inclusion in the meta-analyses, comprising 9 different SNPs: rs280496, rs280500, rs280523, rs280519, rs2304256, rs12720270, rs12720356, rs34536443, and rs35018800. Meta-analysis results showed the minor alleles of rs2304256, rs12720270, rs12720356, rs34536443, and rs35018800 SNPs were associated with protection against autoimmune diseases. Moreover, the A allele of the rs280519 SNP was associated with risk for systemic lupus erythematosus. Our meta-analyses demonstrated that the rs2304256, rs12720270, rs12720356, rs34536443, rs35018800, and rs280519 SNPs in the TYK2 gene are associated with different autoimmune diseases.

ABSTRACT Objective Type 1 diabetes mellitus (T1DM) is an autoimmune disorder caused by a complex interaction between environmental and genetic risk factors. BTB domain and CNC homolog 2 (BACH2) gene encodes a transcription factor that acts on the differentiation and formation of B and T lymphocytes. BACH2 is also involved in the suppression of apoptosis and inflammation in pancreatic beta-cells, indicating a role for it in the development of T1DM. Therefore, the aim of this study was to evaluate the association of the BACH2 rs11755527 single nucleotide polymorphism (SNP) with T1DM. Subjects and methods This case-control study comprised 475 patients with T1DM and 598 nondiabetic individuals. The BACH2 rs11755527 (C/G) SNP was genotyped using real-time PCR with TaqMan MGB probes. Results Genotype distributions of rs11755527 SNP were in accordance with frequencies predicted by the Hardy–Weinberg equilibrium in case and control groups and were similar between groups (P = 0.729). The minor allele frequency was 43.6% in cases and 42.5% in controls (P = 0.604). Moreover, the G allele frequency did not differ between groups when considering different inheritance models and adjusting for age, gender, body mass index, and HLA DR/DQ genotypes of high-risk for T1DM. Although, well-known high-risk T1DM HLA DR/DQ genotypes were associated with T1DM in our population [OR= 7.42 (95% CI 3.34 – 17.0)], this association was not influenced by the rs11755527 SNP. Conclusion The BACH2 rs11755527 SNP seems not to be associated with T1DM in a Brazilian population.

Abstract Uncoupling protein 2 (UCP2) decreases reactive oxygen species (ROS). ROS overproduction is a key contributor to the pathogenesis of diabetic kidney disease (DKD). Thus, UCP2 polymorphisms are candidate risk factors for DKD; however, their associations with this complication are still inconclusive. Here, we describe a case-control study and a meta-analysis conducted to investigate the association between UCP2 -866G/A and Ins/Del polymorphisms and DKD. The case-control study comprised 385 patients with type 1 diabetes mellitus (T1DM): 223 patients without DKD and 162 with DKD. UCP2 -866G/A (rs659366) and Ins/Del polymorphisms were genotyped by real-time PCR and conventional PCR, respectively. For the meta-analysis, a literature search was conducted to identify all studies that investigated associations between UCP2 polymorphisms and DKD in patients with T1DM or type 2 diabetes mellitus. Pooled odds ratios were calculated for different inheritance models. Allele and genotype frequencies of -866G/A and Ins/Del polymorphisms did not differ between T1DM case and control groups. Haplotype frequencies were also similar between groups. Four studies plus the present one were eligible for inclusion in the meta-analysis. In agreement with case-control data, the meta-analysis results showed that the -866G/A and Ins/Del polymorphisms were not associated with DKD. In conclusion, our case-control and meta-analysis studies did not indicate an association between the analyzed UCP2 polymorphisms and DKD.