Satya Narain Sankhwar

Withania somnifera improves semen quality by combating oxidative stress and cell death and improving essential metal concentrations

This study investigated the effect of a 3-month treatment with Withania somnifera on apoptosis and intracellular reactive oxygen species (ROS) concentration of spermatozoa and the metal ions copper, zinc, iron and gold in seminal plasma from infertile men (normozoospermic, n=25; oligozoospermic, n=25; and asthenozoospermic, n=25). The apoptotic and necrotic cell distribution were analysed by annexin-V binding and propidium iodide uptake using flow cytometry. ROS generation was measured by fluorescence intensity and metal ions were analysed by atomic absorption spectrophotometry. The results demonstrated that, prior to treatment, sperm apoptosis and intracellular ROS concentrations were significantly higher in all groups of infertile men compared with controls (P<0.01 to P<0.001). Similarly, the concentrations of the essential metal ions Cu(2+), Zn(2+), Fe(2+) and Au(2+) in seminal plasma were lower. Treatment with W. somnifera significantly reduced apoptosis in normozoospermic and oligozoospermic men and ROS concentrations in oligozoospermic and asthenozoospermic men (all P<0.05). Treatment also significantly improved metal ion concentrations in infertile men (P<0.01). It is concluded that W. somnifera improves semen quality by reducing oxidative stress and cell death, as well as improving essential metal ion concentrations. The aim of this study was to investigate the effect of 3-month treatment with Withania somnifera on apoptosis and intracellular reactive oxygen species (ROS) concentration in spermatozoa from infertile men. Before and following treatment, sperm apoptosis and concentrations of intracellular ROS and the metal ions copper, zinc, iron, and gold in seminal plasma were measured. The apoptotic and necrotic cell distribution were analysed by annexin-V binding and propidium iodide uptake using flow cytometry. ROS generation was measured by fluorescence intensity and metal ions were analysed by atomic absorption spectrophotometry. The results demonstrated that prior, to treatment, apoptosis and intracellular ROS concentrations were significantly higher in all groups of infertile men compared with controls. Similarly, the concentrations of the essential metal ions Cu(2+), Zn(2+), Fe(2+) and Au(2+) in seminal plasma were lower. Treatment with W. somnifera significantly reduced apoptosis and ROS concentrations and improved metal ion concentrations in infertile subjects. It is concluded that W. somnifera improves semen quality by reducing oxidative stress and cell death and improving essential metal ion concentrations.

Metabolomics-derived prostate cancer biomarkers: Fact or fiction?

Despite continuing research for precise probing and grading of prostate cancer (PC) biomarkers, the indexes lack sensitivity and specificity. To search for PC biomarkers, we used proton nuclear magnetic resonance ((1)H NMR)-derived serum metabolomics. The study comprises 102 serum samples obtained from low-grade (LG, n = 40) and high-grade (HG, n = 30) PC cases and healthy controls (HC, n = 32). (1)H NMR-derived serum data were examined using principal component analysis and orthogonal partial least-squares discriminant analysis. The strength of the model was verified by internal cross-validation using the same samples divided into 70% as training and 30% as test data sets. Receiver operating characteristic (ROC) curve examination was also achieved. Serum metabolomics reveals that four biomarkers (alanine, pyruvate, glycine, and sarcosine) were able to accurately (ROC 0.966) differentiate 90.2% of PC cases with 84.4% sensitivity and 92.9% specificity compared with HC. Similarly, three biomarkers, alanine, pyruvate, and glycine, were able to precisely (ROC 0.978) discriminate 92.9% of LG from HG PC with 92.5% sensitivity and 93.3% specificity. The robustness of these biomarkers was confirmed by prediction of the test data set with >99% diagnostic precision for PC determination. These findings demonstrate that (1)H NMR-based serum metabolomics is a promising approach for probing and grading PC.

Low- and High-Grade Bladder Cancer Determination via Human Serum-Based Metabolomics Approach

To address the shortcomings of urine cytology and cystoscopy for probing and grading urinary bladder cancer (BC), we applied (1)H nuclear magnetic resonance (NMR) spectroscopy as a surrogate method for the identification of BC. This study includes 99 serum samples comprising low-grade (LG; n = 36) and high-grade (HG; n = 31) BC as well as healthy controls (HC; n = 32). (1)H NMR-derived serum data were analyzed using orthogonal partial least-squares discriminant analysis (OPLS-DA). OPLS-DA-derived model validity was confirmed using an internal and external cross-validation. Internal validation was performed using the initial samples (n = 99) data set. External validation was performed on a new batch of suspected BC patients (n = 106) through a double-blind study. Receiver operating characteristic (ROC) curve analysis was also performed. OPLS-DA-derived serum metabolomics (six biomarkers, ROC; 0.99) were able to discriminate 95% of BC cases with 96% sensitivity and 94% specificity when compared to HC. Likewise (three biomarkers, ROC; 0.99), 98% of cases of LG were able to differentiate from HG with 97% sensitivity and 99% specificity. External validation reveals comparable results to the internal validation. (1)H NMR-based serum metabolic screening appears to be a promising and less invasive approach for probing and grading BC in contrast to the highly invasive and painful cystoscopic approach for BC detection.

Efficacy of Withania somnifera on seminal plasma metabolites of infertile males: a proton NMR study at 800 MHz.

ETHNOPHARMACOLOGICAL RELEVANCE Traditional Indian systems of medicine use roots of Withania somnifera for impotence, infertility treatment, stress, and the aging process. Although Withania somnifera improves semen quality by regulating reproductive hormone levels and oxidative stress, the molecular mechanism is not clear. AIM OF THE STUDY Our study uses high-resolution Nuclear Magnetic Resonance (NMR) spectroscopy to explore the scientific basis to reveal the pre- and post-treatment efficacy of Withania somnifera on seminal plasma of infertile men-which remains unexplored to date. MATERIALS AND METHODS A total of 180 infertile male patients were administered Withania somnifera root powder at the rate of 5 g/d for a 3-month period. The study included age-matched, healthy men as a control (n=50) group. Proton NMR spectroscopy was used to measure lactate, alanine, glutamate, glutamine, citrate, lysine, choline, glycerophosphocholine (GPC), glycine, tyrosine, histidine, phenylalanine, and uridine in all seminal plasma samples. To appraise infertility levels, we also measured sperm concentration, motility, lipid peroxide, and hormonal perturbation. RESULTS Withania somnifera therapy repairs the disturbed concentrations of lactate, alanine, citrate, GPC, histidine, and phenylalanine in seminal plasma and recovers the quality of semen of post-treated compared to pre-treated infertile men. Serum biochemistry was also improved over post-therapy in infertile men. Our findings reveal that Withania somnifera not only reboots enzymatic activity of metabolic pathways and energy metabolism but also invigorates the harmonic balance of seminal plasma metabolites and reproductive hormones in infertile men. CONCLUSION The results suggest that Withania somnifera may be used as an empirical therapy for clinical management and treatment of infertility.

Low- and high-grade bladder cancer appraisal via serum-based proteomics approach

OBJECTIVE To address the shortcomings of urine cytology and cystoscopy for screening and grading of urinary bladder cancer (BC) we applied a serum-based proteomics approach as a surrogate tactic for rapid BC probing. METHODS This study was performed on 90 sera samples comprising of low-grade (LG, n=33) and high-grade (HG, n=32) BC, and healthy controls (HC, n=25). Two-dimensional gel electrophoresis (2DE) tactic was executed to describe serum proteome. MALDI-TOF-MS (MS) was used to identify the characteristics of aberrantly expressed proteins in 2DE and validated using Western blot (WB) and ELISA approach. Receiver operating characteristics (ROC) curve analysis was also performed to determine the clinical usefulness of these proteins to discriminate among LG, HG and HC cohorts. RESULTS This comprehensive approach of 2DE, MS, WB and ELISA reveals five differentially expressed proteins. Among them two biomarkers (S100A8 and S100A9) were able to accurately (ROC, 0.946) distinguish 81% of BC (LG+HG) cases compared to HC with highest sensitivity and specificity. With a comparable tactic, two biomarkers (S100A8 and S100A4) were able to precisely (ROC, 0.941) discriminate 92% of LG cases from HG with utmost sensitivity and specificity. CONCLUSIONS Serum proteomics probing appears to be an encouraging and least-invasive tactic for screening and grading of BC.

Significant association of TNFα and IL-6 gene with male infertility--an explorative study in Indian populations of Uttar Pradesh.

In this study were aimed to identify the association of SNPs candidate genes of TNF-α and IL-6 with hormones levels and sperm cells death in infertile subjects of Uttar Pradesh population in North India. The study population comprised, fertile donor (control group) and infertile group patients i.e. normozoospermic (idiopathic unexplained), oligozoospermic and asthenozoospermic groups, with 260 subjects in each group. Subjects were selected from the Departments of Urology, K.Gs Medical University and Urology, SGPGIMS, Lucknow, India. The allele-specific polymerase chain reaction (PCR) and PCR-RFLP were used to investigate the substitution of the guanine (G)-to-adenosine (A) at position-308 and guanine (G)-to-cytosine (C) at position-174 in the promoter regions of the TNF-α and IL-6 genes, respectively. Further their relation to male fertility and sperm function were also investigated. It was found that the substitution levels from G to A and from G to C in the TNF-α and IL-6 genes, respectively, were significantly higher in the infertile subjects as compared to that of control group. The apoptosis and necrosis levels were also higher in oligozoospermic and asthenozoospermic infertile subjects. Further it was found to be associated with increased level of reactive oxygen species as observed in oligozoospermic and asthenozoospermic subjects. However, a significant decrease in testosterone and luteinizing hormone with increased prolactin and follicle stimulating hormones was observed in infertile subjects. The study populations indicating a strong association between TNF-α G-308A and IL-6 G-174C substitution with infertile men which is further supported by allele and genotype meta-analysis and thus established it as a risk factor.

NMR spectroscopy of filtered serum of prostate cancer: A new frontier in metabolomics

To address the shortcomings of digital rectal examinations (DRE), serum prostate‐specific antigen (PSA), and trans‐rectal ultrasound (TRUS) for precise determination of prostate cancer (PC) and differentiation from benign prostatic hyperplasia (BPH), we applied 1H‐nuclear magnetic resonance (NMR) spectroscopy as a surrogate tactic for probing and prediction of PC and BPH.

Serum-based protein biomarkers of bladder cancer: A pre- and post-operative evaluation.

Urinary bladder cancer (BC) is the fifth most common cancer worldwide with alarming mortality. Shortcomings of urine cytology and cystoscopy and sparse improvements in the survival rate prompt us to evolve surrogate serum based protein biomarkers to identify BC at an early stage. Previously, we showed that aberrant expression of S100A4, S100A8, S100A9, carbonic anhydrase I (CA I) and Annexin V proteins in pre-operative BC serum compared to healthy controls (HC) (Clin Chim Acta, 2014; 36: 97-103). Here we further evaluate and validate these findings with follow-up post-operative BC patients. This study was conducted on 160 sera samples comprising healthy controls (HC, n=52), pre-operative (n=55) and post-operative (n=53) BC patients. Enzyme-linked immunosorbent assay (ELISA) was used to appraise the aberrantly expressed proteins. ELISA results revealed that the expression levels of S100A8, S100A9, S100A4, and CA I were gradually and significantly reduced; concomitantly, Annexin V was progressively and significantly increased in post-operative compared to pre-operative BC sera samples. Serum protein biomarkers appear to be an encouraging and least-invasive approach for BC identification and prognosticating patient outcomes.

Evaluation of miR-711 as Novel Biomarker in Prostate Cancer Progression

Objective: MicroRNAs (miRs) are class of small non-coding regulatory RNA aberrantly expressed in various types of malignancies including prostate cancer and serves as potential targets to develop new diagnostic and therapeutic strategies. In this quiet we investigated miRNAs expression profile in benign prostatic hyperplasia (BPH) and prostate cancer (PCa) tissue samples and correlated their expression with clinicopathological parameters. Methodology: The miRNAs expression profile as well as their validation has been done by using Microarray and RT-PCR, respectively. Additionally, we also tried to speculate microRNA-mRNA regulatory module through computational target predictions by using Targetscan, Miranda and MirWalk and obtained results were analysed through DAVID software. Result: We observed that miR-711 is significantly deregulated in BPH and PCa, compared to controls. The lower expression of miR-711 was found to be significantly associated with high Gleason score and metastatic disease. Furthermore, the computational target prediction analysis explored miR-711 association to various cancer cells signalling cascade key molecules associated with cancer cell survival. Conclusion: From our observations we suggest that miR-711 may play a critical role in PCa progression, regulation of various cancer cell survival signalling cascades and that it may be a valuable biomarker for prediction of metastatic disease and poor prognosis in PCa.

Relevance of MIC-1 in the Era of PSA as a Serum Based Predictor of Prostate Cancer: A Critical Evaluation

To reduce the ambiguity of contradictory observations in different studies regarding the expression level of Macrophage Inhibitory Cytokine-1 (MIC-1) in serum in prostate cancer (PC), benign prostatic hyperplasia (BPH) and healthy controls (HC), we designed this double-blind study. The study comprises 240 sera from PC, BPH and HC subjects. The expression level of MIC-1 in PC, BPH and HC were appraised using Western blot (WB) and ELISA based approach. WB and ELISA appraisal reveals that the expression level of MIC-1 is significantly higher in PC than in HC or BPH subjects. Regression analysis revealed a significant correlation between MIC-1 vs. PSA (r = 0.09; p < 0.001) and MIC-1 vs. GS (r = 0.7; p < 0.001). ROC analysis using discriminant predicted probability revealed that the MIC-1 was better than PSA. Moreover, the combination of MIC-1 and PSA was allowing 99.1% AUC for the differentiation of BPH + PC from HC, 97.9% AUC for differentiation of BPH from HC, 98.6% AUC for differentiation of PC from HC, and 96.7% AUC for the differentiation of PC from BPH. The augmented expression of MIC-1 in PC compared to BPH and HC subjects is in concurrent of the over-expression of MIC-1 in PC reports and confiscates the contradictory findings of other studies.