INTRODUCTION: It is well known that introduction of intracytoplasmic sperm injection (ICSI) is a breakthrough in the treatment of male infertility. The use of epidydimal and testicular sperm has been beneficial in the treatment of azoospermic men. To identify the impact of surgically retrieved sperm (SRS) on reproductive and peri-natal outcomes in Assisted Reproductive Treatment (ART) cycles. This is a retrospective study design at our private IVF clinic duration is from 2009 to 2019. The sample size of this study is a total of 989 subjects underwent SRS under general anaesthesia. Of these only 552 couples were included in this study with female partner's age ≤ 37 and had self-gamete cycles. Among them only 342 subjects (till June 2019) had embryo transfer. Couples with ejaculate sperm (ES) and self-oocytes who had ICSI cycles acted as control group for this study (n=425). In study group male partners underwent SRS and female partner's underwent controlled ovarian stimulation and trans vaginal oocyte retrieval as per our clinic's standard operating protocols (SOP). Embryos were cultured till blastocysts and a freeze all policy was followed. Frozen Embryo transfer with two good grade blastocysts (grading done as per Istanbul Consensus by Alpha scientists) was done in subsequent cycles. Primary Outcomes: Live birth rates (LBR) and Peri-Natal outcomes- Obstetric complications (Gestational diabetes (GDM), Pregnancy Induced Hypertension (PIH), Intra-Uterine Growth Restriction (IUGR), and foetal anomalies. Secondary Outcomes: Reproductive out comes – Fertilization rate (FR); blastocyst rate (BR); Implantation rate (IR); Clinical pregnancy rate (CPR); Miscarriage rate (MR) and Multiple Pregnancy rates (MPR). Mean Reproductive and Perinatal outcomes of Study Vs Control group are as follows: FR – 92.6% Vs 92.3% (P=0.8760) BR – 37.18% Vs 48.4% (P=0.0018) CPR – 58.18 Vs 62.93% (P=0.1807) IR – 40.63% Vs 48.97% (P=0.0212) MR- 7.02% Vs 9.98% (P=0.1477) MPR – 10.81% Vs 17% (P=0.0148) LBR – 40.06% Vs 51.4% (P=0.0018) GDM – 3.8% Vs 1.8% (P=0.0891) PIH – 2.63% Vs 2.12% (P=0.6431) IUGR – 0% Vs 0.71% Lost to Follow up – 10% Vs 0% (P=0.8760) Congenital Anomalies – One baby with Angleman's Syndrome in study groups Vs one baby with hydrocephalus, one death due to Respiratory distress and 3 deaths in NICU after preterm birth in control group. Data from this study is suggesting that use of SRS seems to offer beneficial reproductive and perinatal outcomes. Though Blastocycst rate and LBR is significantly higher in control group, SRS groups still has acceptable outcomes. CONCLUSIONS:Use of SRS in ART cycles do not seem to alter reproductive and peri-natal outcomes and couples should be reassured and counselled appropriately. However, long term follows up of the health of such off springs is still warranted.

To evaluate reproductive outcomes of Advanced Paternal Age (APA) men using Surgically Retrieved Testicular Sperm (SRS) in Assisted Reproductive Technology (ART) cycles. Retrospective Observational Longitudinal Cohort study In Private Teaching ART centre. Men over the age of 40 were classified as APA and recruited in this study. Age range of male partners was in between 40-50 years. Couples where women were young (<37yrs age) and male partners underwent SRS in ART cycles were considered for this study during January 2014 to July 2019 (n=103). Couples undergoing self-gamete cycles were considered for the study and couples undergoing PGT-A were excluded from the study. APA men & Younger women in whom ejaculated sperm was used in ART cycles acted as control group for this study (n=71). All women underwent controlled ovarian stimulation and Transvaginal oocyte retrieval (OPU). Male partner's underwent either fresh SRS on OPU day or frozen SRS was used for ICSI as per our clinics standard operating procedures (SOP). Embryos were cultured till blastocyst stage and a freeze all policy was adopted. Two Blastocysts were transferred in a Frozen Embryo Replacement Cycle (FET). Reproductive outcomes were compared between the two groups. The reproductive outcomes for Study Vs Control group were as follows: Clinical Pregnancy Rate – 45.71% Vs 56.16% (p value 0.1767) Implantation Rates -33.6% Vs 42.15% (p value 0.2525) Miscarriage Rates – 15.71% vs 8.22% (p value 0.1457) Live Birth Rates – 27.14% vs 53.33% (p value 0.0005) Multiple Pregnancy rate – 21.42% Vs 5.48% (p value 0.0038) Data from this study is suggestive that, testicular sperms from APA men could alter reproductive outcomes in ART cycles. APA men undergoing ART cycles with SRS need to be counselled appropriately about the limitations. Role of PGT-A in such group of men to optimize reproductive outcomes needs further research. Testicular Sperm of advanced paternal age men seems to have detrimental effects on reproductive outcomes.

To investigate Reproductive & Perinatal Outcomes of Fresh or Frozen-thawed surgically retrieved sperms (SRS) in Non-Obstructive Azoospermia (NOA) individuals Retrospective data Analysis of (n=108) patients undergoing SRS during January 2013 to June 2019.The study group was divided into two groups, Fresh SRS (n=60) and Frozen SRS (n=48) in our private IVF clinic. Study Design and Duration-Retrospective data Analysis of (n=108) patients undergoing SRS during January 2013 to June 2019.The study group was divided into two groups, Fresh SRS (n=60) and Frozen SRS (n=48) in our private IVF clinic. Inclusion-Couples undergoing Assisted Reproductive Technique (ART) cycles where the women were < 37 years old and male partners had NOA and had a successful retrieval of sperm, were included. Only self-gamete cycles were considered. Exclusion –Advanced maternal age (>38yrs), People with no sperm retrieval, People who did not have viable sperms after freezing and thawing. Women underwent controlled ovarian stimulation and oocyte retrieval (OPU) as per our clinic's standard operating procedure (SOP). Male partner's underwent SRS and testicular sperms were either used for ICSI if done at OPU or frozen and thawed for use in subsequent cycles as per SOP. Embryos cultured till blastocyst stage and freeze all policy was adopted. Couples underwent Frozen embryos transfers with two blastocysts and reproductive outcomes were evaluated. Primary Outcome- Blastocyst Rate (BR), Implantation Rate (IR), Live Birth Rate (LBR). Secondary Outcome-Fertilization Rate (FR), Miscarriage Rate (MR), Clinical Pregnancy Rate (CPR), Perinatal Outcomes. The Reproductive outcome for Fresh and Frozen Surgically Retrieved Sperms, respectively were, Fertilization Rate was 92.72 % v/s 92.74 % (p value 0.9968) Blastocyst Rate was 46.74 % v/s 35.57 % (p value 0.2444) Implantation Rate was 43.97 % v/s 47.27 %(p value 0.7334) Clinical Pregnancy Rate was 59.67 % v/s 73.33 % (p value 0.1390) Live Birth Rate was 45.16 % v/s 63.33 % (p value 0.0612) Miscarriage Rate was 4.84 % v/s 3.33 % (p value 0.6978) Perinatal Outcomes Both study groups had comparable perinatal outcomes with no concerning trends. Congenital anomalies - Only One baby from fresh SRS had Anglemans syndrome. There isn't a marked difference in the outcomes between Fresh or Frozen SRS. Freezing testicular sperms doesn't seem to alter reproductive and perinatal outcomes. Frozen Testicular Sperm seem to offer comparable outcomes with Fresh Testicular sperms in ART cycles. All trial attempts for sperm retrieval should be backed up with cryopreservation.

For Individuals with Raised Sperm DNA Fragmentation (SDF), will intervention with Surgical Retrieval of Sperms (SRS) from testes optimize Reproductive and Peri-Natal outcomes in Assisted Reproductive Techniques (ART) cycles. This is a retrospective study at our private IVF clinic from 2010 to 2019. SAMPLE SIZE: Couples with history of at least one failed IVF cycle were offered testing for SDF by SCSA method. SDF >30% was considered high and such couples were offered SRS from testes and testicular sperms were used for ICSI (n= 342). Couples were counselled about the risks, benefits and experimental nature of this intervention. Written consent was obtained from the couple and taken through an ART cycle with SRS from tests. Women underwent controlled ovarian stimulation and oocyte retrieval as per our clinic's standard operating protocol (SOP). Fertilized oocytes were cultured till blastocyst stage and freeze all policy was adopted. Two blastocysts were transferred in a frozen embryo replacement cycle. INCLUSION CRITERIA: Individuals who had successful SRS with self-gamete cycles were included in this study. Exclusion CRITERIA: Donor gamete cycles and individuals who had failed SRS were excluded from this study. Primary Outcomes: Implantation rate (IR), Live birth rates (LBR) and Perinatal outcomes (Gestation Diabetes (GDM), Pregnancy Induced Hypertension (PIH), Intra Uterine Growth Restriction (IUGR), mean Pre-Term Birth rate (PTB), Birth weight (BW Normal - >2.5kgs), Low Birth weight (LBW – 1.5-2.5kgs), Very LBW (VLBW - <1.5kgs) Secondary Outcomes: Fertilization rate (FR); blastocyst rate (BR), Clinical pregnancy rate (CPR); Miscarriage rate (MR) and Multiple Pregnancy rate (MPR) Mean of the outcomes were as follows: FR - 93.35%, BR - 35%, IR - 40.34%, CPR - 56.90 %, MR was 12.03 %, MPR – 17.2% , LBR - 35.23, GDM – 6.06%, PIH – 6.82%, No IUGR reported in the study group. 17.4% had PTB. Mean BW was 2.5 kg. Out of this Normal BW was 55.6 %, LBW was 36.7 % and VLBW was 13.3 %. Use of SRS in couples with raised SDF seems to offer optimal reproductive and perinatal outcomes. Couples seeking ART with raised SDF, SRS can be offered as an active intervention. Testicular sperm for Raised SDF seem to offer beneficial reproductive and perinatal outcomes in ART cycles. TESA can be offered as an active intervention.

To look at the perinatal outcome of surgically retrieved spermatozoa (SRS) in individuals with Non obstructive Azoospermia (NOA). This is a Retrospective Study at our Private IVF clinic from 2010 to 2019. 989 patients underwent SRS under general anaesthesia. Of these 264 patients were indicated for NOA. 155 patients had successful Sperm retrieval and 93 patients finally had successful frozen embryo transfers with two good grade blastocysts and 425 ICSI cycles with Ejaculate sperms (EjS) were in control group. In this study couples who had successful SRS from testes and Intra-Cytoplasmic Sperm Injection (ICSI) with self-Oocytes and had blastocysts frozen for transfer were included. All NOA patients underwent SRS either before the oocyte retrieval (OPU) and frozen for subsequent use or SRS was done at OPU and fresh testicular sperms used for ICSI. SRS was done as per our clinics standard operating protocol (SOP). All women underwent controlled ovarian stimulation, OPU and ICSI with fresh or frozen SRS. Embryos cultured till blastocyst stage and a freeze all policy was followed. Two blastocysts were transferred in a frozen cycle. PRIMARY OUTCOMES – Live Birth rate (LBR) and Perinatal outcomes SECONDARY OUTCOMES – Reproductive Outcome Fertilization rate (FR), Blastocyst rate (BR), Implantation Rate (IR), clinical pregnancy rate (CPR), Miscarriage rate (MR), Multiple Pregnancy Rate(MPR) and Lost for follow up (LFU). Mean outcomes of Study Vs Control group were as follows: FR - 92.35% Vs 92.30% (P= 1.0000) BR - 42.59% Vs 48.8% (P=0.2779 ) CPR - 64.52% Vs 62.93% (P= 0.7735) IR - 45.35% Vs 48.97% (P= 0.5273) MR -7.53% Vs 9.98% (P=0.4667 ) LFU is 9.78% Vs 0% LBR - 46.47% Vs 51.4% (P=0.3895 ) MPR -27.27% Vs 17% (P=0.0219 ) Maternal complications Gestational Diabetic Mellitus - 3% Vs 1.8% (P= 0.4562) Pregnancy Induced Hypertension (PIH) – 0% Vs 2.1% (P=0.1590 ) Intra-Uterine Growth Restriction (IUGR) – 0% Vs 0.3% (P= 0.5973) Perinatal outcomes – Average BW - 2.52 Kg Vs 3.01 kg %Normal BW (>2.5kgs)– 59.52% Vs 63% (P= 0.5307) % Low BW (1.5-2.5kgs)- 30.95% Vs 31% (P=0.9925) %Very Low BW (<1.5kgs) - 9.52% Vs 6% (P= 0.2170) pre-term Birth (<37weeks gestation) - 15.5% Vs 12.8% (P= 0.4876) Congenital Anomalies in Study Group one baby was diagnosed with Angelman syndrome (AS), one infant diagnosed with Rabdomyoma at 5th month in study group Congenital Anomalies in Control Group one baby with hydrocephalus, one ARDS due to meconium aspiration and the baby died after 18 days, 3 babies died in NICU after pre term delivery. Data from this study doesn't seem to adversely affect Reproductive and Perinatal outcomes with use of Testicular sperms for ICSI in NOA men. In NOA use of SRS doesn't seem to alter reproductive and perinatal outcomes. However, long term follow up of these children is still advised.

PURPOSE:The rationale of the current study was to develop 6-mercaptopurine (6-MP)-mediated hematological toxicity prediction model for acute lymphoblastic leukemia (ALL) therapeutic management.METHODS:A total of 96 children with ALL undergoing therapy with MCP-841 protocol were screened for all the ten exons of TPMT, exon 2, exon 3 and intron 2 of ITPA using bidirectional sequencing. This dataset was used to construct prediction models of leucopenia grade by constructing classification and regression trees (CART) followed by smart pruning.RESULTS:The developed CART model indicated TPMT*12 and TPMT*3C as the key determinants of toxicity. TPMT int3, int4 and int7 polymorphisms exert toxicity when co-segregated with one mutated allele of TPMT*12 or TPMT*3C or ITPA exon 3. The developed CART model exhibited 93.6% accuracy in predicting the toxicity. The area under the receiver operating characteristic curve was 0.9649.CONCLUSIONS:TPMT *3C and TPMT*12 are the key determinants of 6-MP-mediated hematological toxicity while other variants of TPMT (int3, int4 and int7) and ITPA ex2 interact synergistically with TPMT*3C or TPMT*12 variant alleles to enhance the toxicity. TPMT and ITPA variants cumulatively are excellent predictors of 6-MP-mediated toxicity.

Effective immunosuppression through optimization of trough levels tacrolimus reduces post-transplant mortality rate in liver transplant cases. Meta-analysis was carried out to evaluate how donor/recipient CYP3A5 (n = 678) and recipient ABCB1 (n = 318) genotypes influence tacrolimus pharmacokinetics till one-month of transplantation. The donor CYP3A5*3/*3 genotype exhibited higher concentration/dose (C/D) ratio of tacrolimus in week 1 (mean difference: 65.04, 95% CI: 15.30–114.79 ng/ml/mg/kg), week 2 (mean difference: 21.7, 95% CI: 12.6–30.9 ng/ml/mg/kg) and week 4 (mean difference: 43.28, 95% CI: 17.09 — 69.49 ng/ml/mg/kg) compared to *1/*1 and *1/*3 genotypes. The recipient CYP3A5 *3/*3 genotype did not showed significant difference in tacrolimus C/D ratio in week 1 compared to other two genotypes. However, week 2 (mean difference: 44.16, 95% CI: 3.68–84.65 ng/ml/mg/kg) and week 4 (mean difference: 43.74, 95% CI: 12.50–75.00 ng/ml/mg/kg) availability was higher in *3/*3 mutant recipients. However, the recipient ABCB1 3435 C > T polymorphism has no significant influence on tacrolimus pharmacokinetics till one month of transplant. The donor and recipient CYP3A5*3 polymorphism influences tacrolimus pharmacokinetics in the first month post-transplantation, whereas the association with recipient ABCB1 3435 C > T is inconclusive.

INTRODUCTION:In view of our previous studies showing an independent association of genetic polymorphisms in folate, xenobiotic, and toll-like receptor (TLR) pathways with the risk for systemic lupus erythematosus (SLE), we have developed three statistical models to delineate complex gene-gene interactions between folate, xenobiotic, TLR, and signal transducer and activator of transcription 4 (STAT4) signaling pathways in association with the molecular pathophysiology of SLE.METHODS:We developed additive, multifactor dimensionality reduction (MDR), and artificial neural network (ANN) models.RESULTS:The additive model, although the simplest, suggested a moderate predictability of 30 polymorphisms of these four pathways (area under the curve [AUC] 0.66). MDR analysis revealed significant gene-gene interactions among glutathione-S-transferase (GST)T1 and STAT4 (rs3821236 and rs7574865) polymorphisms, which account for moderate predictability of SLE. The MDR model for specific auto-antibodies revealed the importance of gene-gene interactions among cytochrome P450, family1, subfamily A, polypeptide 1 (CYP1A1) m1, catechol-O-methyltransferase (COMT) H108L, solute carrier family 19 (folate transporter), member 1 (SLC19A1) G80A, estrogen receptor 1 (ESR1), TLR5, 5-methyltetrahydrofolate-homocysteine methyltransferase reductase (MTRR), thymidylate synthase (TYMS). and STAT4 polymorphisms. The ANN model for disease prediction showed reasonably good predictability of SLE risk with 30 polymorphisms (AUC 0.76). These polymorphisms contribute towards the production of SSB and anti-dsDNA antibodies to the extent of 48 and 40%, respectively, while their contribution for the production of antiRNP, SSA, and anti-cardiolipin antibodies varies between 20 and 30%.CONCLUSION:The current study highlighted the importance of genetic polymorphisms in folate, xenobiotic, TLR, and STAT4 signaling pathways as moderate predictors of SLE risk and delineates the molecular pathophysiology associated with these single nucleotide polymorphisms (SNPs) by demonstrating their association with specific auto-antibody production.

Cytotoxic T lymphocyte associated-antigen (CTLA4) is a potential negative regulatory molecule of T-cells and associated with several autoimmune diseases. Several reports from different ethnic groups showed that the polymorphisms of the CTLA4 gene have been associated with autoimmune diseases including SLE. Therefore, we aimed to investigate the +49 A/G polymorphism in South Indian SLE patients and its association with disease aetiology and serological markers. A total of 534 samples were genotyped for the +49 A/G polymorphism in exon 1 of the CTLA-4 gene through PCR-RFLP method. We found significant association of genotype and allele frequencies with +49 A/G polymorphism in SLE patients. The frequency of the +49 A/G polymorphism rs231775 'GG' genotype was significantly higher in patients with SLE (12.32%) than those in healthy control subjects (4.6%) (OR: 1.797; 95% CI 1.264-2.554; p=0.001). The frequency of mutant allele 'G' also found to be significantly higher in cases (36.01%) than controls (24.92%) (OR: 1.695, 95% CI: 1.298-2.214, p<0.001). We observed significant increase in serum TNF-α, interferon-α, IL-10 and IL-12 in SLE cases compared to controls. We also found a significant association of serum TNF-α, interferon-α, IL-10 and IL-12 with SLE phenotypes. In addition there was a significant increase in serum TNF-α level in "GG" genotype SLE subjects suggesting that it might play a major role in the advancement of SLE disease.

The present work was aimed to study the association of one carbon genetic variants, hyperhomocysteinemia and oxidative stress markers, i.e., serum nitrite, plasma malondialdehyde (MDA) and glutathione (GSH) on intimal medial thickening (IMT) in patients with type 2 diabetes mellitus (T2D). A total number of 76 subjects from ACS Medical College and Hospital, Chennai, India were included in the study, i.e., Group I (n = 42) of T2D and Group II (n = 34) of age- and sex matched healthy controls. The glycated haemoglobin was measured by ion-exchange resin method; plasma homocysteine by Enzyme Linked Immunosorbant Assay method; serum nitrite (nitric oxide, NO), plasma MDA and GSH by spectrophotometric methods; the IMT by high frequency ultrasound. The polymorphisms of one carbon genetic variants were genotyped using polymerase chain reaction-restriction fragment length polymorphism and amplified fragment length polymorphism methods. Results indicate that methyltetrahydrofolate homocysteine methyl transferase (MTR) A2756G allele was found to be protective in T2D and the other variants were not significantly associated with T2D. Glutamate carboxypeptidase II (GCP II) C1561T (r = 0.34; p = 0.05) and methylene tetrahydrofolate reductase (MTHFR) C677T (r = 0.35; 0.04) showed positive correlation with plasma homocysteine in T2D cases. In this study, MTR A2756G allele was found to be protective in T2D; GCP II C1561T and MTHFR C677T showed positive association with plasma homocysteine in T2D cases. Among all the genetic variants, MTR A2756G was found influence IMT. RFC 1 G80A and TYMS 5′-UTR 2R3R showed synergistically interact with MTR A2756G in influencing increase in IMT.

Putatively functional polymorphisms of one-carbon and xenobiotic metabolic pathways influence susceptibility for wide spectrum of diseases. The current study was aimed to explore gene-gene interactions among these two metabolic pathways in four diseases i.e. breast cancer, systemic lupus erythematosus (SLE), coronary artery disease (CAD) and Parkinson's disease (PD). Multifactor dimensionality reduction analysis was carried out on four case-control datasets. Cross-talk was observed between one-carbon and xenobiotic pathways in breast cancer (RFC 80 G>A, COMT H108L and TYMS 5'-UTR 28 bp tandem repeat) and SLE (CYP1A1 m1, MTRR 66 A>G and GSTT1). Gene-gene interactions within one-carbon metabolic pathway were observed in CAD (GCPII 1561 C>T, SHMT 1420 C>T and MTHFR 677 C>T) and PD (cSHMT 1420 C>T, MTRR 66 A>G and RFC1 80 G>A). These interaction models showed good predictability of risk for PD (The area under the receiver operating characteristic curve (C) = 0.83) and SLE (C = 0.73); and moderate predictability of risk for breast cancer (C = 0.64) and CAD (C = 0.63). Cross-talk between one-carbon and xenobiotic pathways was observed in diseases with female preponderance. Gene-gene interactions within one-carbon metabolic pathway were observed in diseases with male preponderance.

The rationale of this case-control study was to explore the association of Toll-like receptor 4 (TLR4) D299G, TLR4 T399I, TLR9 -1486 T>C, TIR-domain-containing adaptor protein (TIRAP) S180L and tumor necrosis- (TNF-) promoter polymorphisms with susceptibility and phenotypic heterogeneity of systemic lupus erythematosus (SLE). PCR-RFLP, real-time PCR was used for the genetic analysis and expression studies and ELISA was used for the determination of specific autoantibodies. TLR4 D299G was associated with the risk for SLE (OR: 1.57, 95% CI: 1.08-2.28), while the TNF- (-1031, -863, -857) CCC haplotype conferred protection. TLR4 and TIRAP polymorphisms were associated with reduced expression of HLA-DR. The presence of TLR4 and TLR9 polymorphisms increases the MHC2TA expression, while TIRAP polymorphism was associated with reduced expression. TLR4 D299G showed an inverse association with pulmonary hypertension. TLR 4 T399I and TLR9 -1486T>C showed a positive association with seizures and photosensitivity, respectively. TIRAP S180L showed a positive association with alopecia and malar rashes, while an inverse association with psychosis was observed. TLR4 T399I (r=0.14, p=0.05) and TIRAP S180L (r=0.15, p=0.03) showed a positive association with anti-Ro antibodies. On the other hand, TLR9 -1486T>C showed an inverse association with anti-La antibodies (r=-0.20, p=0.006). To conclude, TLR4 D299G increases the risk for SLE, while TNF- CCC haplotype reduces the risk for SLE. All these polymorphisms contribute toward phenotypic heterogeneity. TLR4 T399I, TLR9 -1486T>C and TIRAP S180L influence specific autoantibody production in SLE.

Systemic lupus erythematosus (SLE) is a complex autoimmune disorder involving genetic, epigenetic and environmental factors and has higher incidence in women. In this study, we explored the association of estrogen receptor 1 (ESR1) rs2234693 (PvuII) and rs9340799 (XbaI) polymorphisms with susceptibility to SLE. PCR-RFLP and ELISA were used for genetic analysis and determination of specific autoantibodies, respectively. The univariate analysis showed no independent association of rs2234693 (OR: 1.14, 95% CI: 0.87 - 1.49, p = 0.36) and rs9340799 (OR: 0.87, 95% CI: 0.66-1.14, p = 0.34). The haplotype analysis using SHEsis platform revealed strong linkage disequilibrium between these two polymorphisms (D': 0.81, r2: 0.55). Among the four haplotype groups, the C-A haplotype (rs2234693-rs9340799) was strongly associated with the risk for SLE (OR: 2.10, 95% CI: 1.32 - 3.34, p = 0.001). The homozygous variant genotype of rs2234693 exhibited elevated TNF-α and depleted IFN-α, while the effects of rs9340799 were contradictory. The wild genotype of rs2234693 exhibited lower levels of IL-12 with number of rs9340799 variant alleles pronouncing this effect. From this study, it is concluded that the ESR1 haplotypes may influence the Th2 cytokine profile and susceptibility to SLE among the South Indians.

The current study was conducted to elucidate the effect of genetic variations in one-carbon metabolism on the epigenetic regulation of major histocompatibility complex II transactivator (MHC2TA), reduced folate carrier 1 (RFC1/SLC19A1) and human leukocyte antigen (HLA)-DR in systemic lupus erythematosus (SLE). PCR-RFLP/AFLP, bisulfite-sequencing and real-time PCR approaches were used for genetic, epigenetic and expression analysis respectively. SLE cases exhibited elevated plasma homocysteine levels compared to healthy controls (24.93 ± 1.3 vs. 11.67 ± 0.48 μmol/l), while plasma folate levels showed no association (7.10 ± 2.49 vs. 7.64 ± 2.09 ng/ml). The RFC1 80G>A polymorphism showed 1.32-fold risk (95% CI: 1.02-1.72) for SLE, while glutamate carboxypeptidase II (GCPII) 1561C>T showed reduced risk (OR: 0.47, 95% CI: 0.24-0.90). The expression of RFC1 (0.37 ± 0.09 vs. 0.60 ± 0.17) and HLA-DR (0.68 ± 0.17 vs. 0.98 ± 0.02) was down regulated in the SLE cases. The hypermethylation of RFC1 as observed in the current study may contribute for its down regulation. Plasma folate and thymidylate synthase (TYMS) 5'-UTR 28 bp tandem repeat showed an inverse association with methylation of RFC1 and MHC2TA. SLE cases with hypocomplementemia showed hypermethylation of RFC1, hypomethylation/up regulation of MHC2TA and down regulation of HLA-DR. The hypermethylation of MHC2TA and down regulation of RFC1, MHC2TA and HLA-DR were observed in anti-cardiolipin antibody positive SLE cases. The up regulation of RFC1 and HLA-DR was observed in anti-dsDNA antibody positive SLE cases. The hypomethylation/upregulation of RFC1 and MHC2TA was observed in anti-RNP antibody positive cases. To conclude, one-carbon genetic variants influence epigenetic of MHC2TA and RFC1, thus contributing to phenotypic heterogeneity of SLE.

In view of documented evidence that catechol estrogen-DNA adducts serve as epitopes for binding of anti-nuclear antibodies, genetic polymorphisms of the xenobiotic metabolic pathway involved in estrogen metabolism might contribute towards pathophysiology of systemic lupus erythematosus (SLE). To test this hypothesis, a case-control study was conducted. Cytochrome P 450 1A1 (CYP1A1) m4 (OR: 4.93, 95% CI: 1.31-18.49), catecholamine-o-methyl transferase (COMT) H108L (OR: 1.39, 95% CI: 1.03-1.88) and glutathione-S-transferase (GST) T1 null (OR: 1.83, 95% CI: 1.11- 3.01) variants showed association with SLE risk. SHEsis web-based platform analysis showed mild to moderate linkage disequilibrium between the CYP1A1 ml, m2 and m4 variants (D': 0.19-0.37). Among the different haplotypes of CYP1A1, CAC-haplotype harboring CYP1A1 ml variant showed association with SLE risk (OR: 1.46, 95% CI: 1.11-1.92). Multifactor dimensionality reduction analysis (MDR) showed potential gene-gene interactions between the phase II variants i.e. COMT H108L x GSTT1 null x GSTM1 null (p < 0.0001) and also between the phase II and I variants i.e. COMT H108L x GSTTI null x CYP1A1 ml x CYP1A1 m2 in inflating the risk of SLE by 3.33-folds (95% CI: 2.30-4.82) and 4.00-folds (95% CI: 2.77-5.78), respectively. To conclude, hyperinducibility of CYP1A1 due to ml and m4 variants and defective phase-II detoxification due to COMT H108L and GSTT1 null variants increase the susceptibility to SLE.

The aim of this case–control study is to explore the role of aberrations in xenobiotic metabolism in inducing oxidative DNA damage and altering the susceptibility to breast cancer. Cytochrome P4501A1 (CYP1A1) m1 (OR: 1.41, 95% CI 1.08–1.84), CYP1A1 m4 (OR: 5.13, 95% CI 2.68–9.81), Catecholamine-O-methyl transferase (COMT) H108L (OR: 1.49, 95% CI 1.16–1.92), and glutathione S-transferase (GST) T1 null (OR: 1.68, 95% CI 1.09–2.59) variants showed association with breast cancer risk. Reduced folate carrier 1 (RFC1) 80A/CYP1A1 m1/CYP1A1 m4 and RFC1 80A/thymidylate synthase (TYMS) 5′-UTR 2R/methionine synthase (MTR) 2756G/COMT 108L genetic combinations were found to inflate breast cancer risk under the conditions of low dietary folate (345 ± 110 vs. 379 ± 139 μg/day) and low plasma folate (6.81 ± 1.25 vs. 7.09 ± 1.26 ng/ml) by increasing plasma 8-oxo-2′-deoxyguanosine (8-oxodG). This increase in 8-oxodG is attributed to low methionine (49.38 ± 23.74 vs. 53.90 ± 23.85 μmol/l); low glutathione (378 ± 242 vs. 501 ± 126 μmol/l) and GSTT1 null variant; and hypermethylation of CpG island of extracellular-superoxide dismutase (EC-SOD) (92.78 ± 11.49 vs. 80.45 ± 9.86%), which impair O-methylation of catechol estrogens to methoxy estrogens, conjugation of glutathione to semiquinones/quinones and free radical scavenging respectively. Our results suggest cross-talk between one-carbon metabolism and xenobiotic metabolism influencing oxidative DNA damage and susceptibility to breast cancer.

To optimize the warfarin dose, a population-specific pharmacogenomic algorithm was developed using multiple linear regression model with vitamin K intake and cytochrome P450 IIC polypeptide9 (CYP2C9*2 and *3), vitamin K epoxide reductase complex 1 (VKORC1*3, *4, D36Y and −1639 G>A) polymorphism profile of subjects who attained therapeutic international normalized ratio as predictors. New algorithm was validated by correlating with Wadelius, International Warfarin Pharmacogenetics Consortium and Gage algorithms; and with the therapeutic dose (r=0.64, P<0.0001). New algorithm was more accurate (Overall: 0.89 vs 0.51, warfarin resistant: 0.96 vs 0.77 and warfarin sensitive: 0.80 vs 0.24), more sensitive (0.87 vs 0.52) and specific (0.93 vs 0.50) compared with clinical data. It has significantly reduced the rate of overestimation (0.06 vs 0.50) and underestimation (0.13 vs 0.48). To conclude, this population-specific algorithm has greater clinical utility in optimizing the warfarin dose, thereby decreasing the adverse effects of suboptimal dose.

The authors investigated the role of dietary micronutrients and eight functional polymorphisms of one-carbon metabolism in modulating oxidative stress in sporadic breast cancer. PCR-restriction fragment length polymorphism (RFLP) and PCR-amplified fragment length polymorphism (AFLP) methods were used for genetic analysis in 222 sporadic breast cancer cases and 235 controls. Standardized food frequency questionnaire was used for dietary micronutrient assessment. 8-oxo-2′-deoxyguanosine (8-oxodG), folate, and estradiol were estimated using commercial ELISA kits. Reverse-phase HPLC coupled with fluorescence detector was used for plasma homocysteine analysis. Total glutathione was estimated using Ellman’s method. Reduced folate carrier 1 (RFC1) G80A and methylenetetrahydrofolate reductase (MTHFR) C677T were associated with risks of 1.34 (95% CI 1.01–1.79)- and 1.84 (95% CI 1.14–3.00)-folds, respectively, for sporadic breast cancer while cytosolic serine hydroxymethyl transferase (cSHMT) C1420T was associated with reduced risk (OR 0.71, 95% CI 0.53–0.94). Significant increase in plasma 8-oxo-2′-deoxyguanosine (P < 0.004) and homocysteine (P < 0.0001); and significant decrease in total glutathione (P < 0.01) and dietary folate (P = 0.006) was observed in cases than in controls. Oxidative DNA damage showed direct association with menopause (P = 0.02), RFC1 G80A (P < 0.05) and homocysteine (P < 0.0001); and inverse association with dietary folate (P < 0.0001), plasma folate (P < 0.0001), cSHMT C1420T (P < 0.05) and glutathione (P < 0.001). To conclude, the aberrations in one-carbon metabolism induce oxidative stress in sporadic breast cancer either by affecting the folate pool or by impairing remethylation.

AIM:The current study aimed to address the inconsistencies in association studies, specifically with reference to methylene tetrahydrofolate reductase (MTHFR) C677T polymorphism in the light of gene-gene and gene-nutrient interactions.METHODS:A case-control study was conducted to analyze four genetic polymorphisms i.e. thymidylate synthase (TYMS) 5'-UTR 28 bp tandem repeat, MTHFR C677T, methyltetrahydrofolate homocysteine methyltransferase (MTR) A2756G, methyltetrahydrofolate homocysteine methyltransferase reductase (MTRR) A66G using PCR-AFLP and PCR-RFLP methods; plasma folate and B12 using AxSYM kits; plasma homocysteine by reverse phase HPLC and nitric oxide using Griess reaction. Fisher's exact test, logistic regression analysis and multifactor dimensionality reduction analysis were used for statistical analysis of genetic parameters. Student's t-test was used for biochemical parameters.RESULTS:MTHFR C677T and MTRR A66G were found to increase the risk for CAD by 1.61-fold (95% CI: 1.04-2.50) and 1.92-fold (95% CI: 1.29-2.87) whereas TYMS 2R allele was found to reduce the risk for CAD (OR: 0.66, 95% CI: 0.49-0.88) by counteracting MTHFR and MTRR variant alleles. Significant gene-gene interactions were observed among TYMS/MTRR (P < 0.0001), MTR/TYMS/MTRR (P < 0.0001), and MTHFR/MTR/TYMS/MTRR (P < 0.0001). MTHFR was found to increase the risk (OR: 2.36, 95% CI: 1.28-4.37) only in the absence of the TYMS 2R allele, with marked impairment of the remethylation process (P = 0.007). This impairment was predominant when the dietary folate was in the lowest tertile. In subjects with dietary folate intake in the highest tertile, no such impairment was observed.CONCLUSION:Dietary folate status and TYMS 5'-UTR 28bp tandem repeat polymorphism are important effect modifiers of CAD risk associated with genetic variants in remethylating genes.