Environmental & lifestyle factors in deterioration of male reproductive health.

BACKGROUND & OBJECTIVES: Male reproductive function in the general population has been receiving attention in recent years due to reports of various reproductive and developmental defects, which might be associated with various lifestyle and environmental factors. This study was carried out to determine the role of various lifestyle and environmental factors in male reproduction and their possible association with declining semen quality, increased oxidative stress as well as sperm DNA damage. METHODS: Semen samples were obtained from 240 male partners of the couples consulting for infertility problem. Semen analysis was carried out using WHO criteria and subjects were categorized on the basis of self reported history of lifestyle as well as environmental exposure. The oxidative and antioxidant markers; lipid peroxidation (LPO), superoxide dismutase (SOD) and catalase (CAT) as well as DNA damage by acridine orange test (AO) were determined. RESULTS: The presence of abnormal semen parameters was significantly higher among the lifestyle and/or environmental exposed subjects as compared to the non-exposed population. Further, the levels of antioxidants were reduced and sperm DNA damage was more among the lifestyle and/or environmental exposed subjects, though the changes were not significant. INTERPRETATION & CONCLUSIONS: These findings indicated that various lifestyle factors such as tobacco smoking, chewing and alcohol use as well as exposure to toxic agents might be attributed to the risk of declining semen quality and increase in oxidative stress and sperm DNA damage.

Successful generation of donor specific hematopoietic stem cell lines from co-cultured bone marrow with human embryonic stem cell line: a new methodology.

We report the generation of 30 healthy human embryonic stem cell (h-ESC) lines from 33 voluntary oocyte donors using a donor somatic cell nuclear transfer (SCNT) technique on 190 oocytes. Our aim was to coculture them with their own bone marrow (BM) to generate hematopoietic progenitor cells for therapeutic purposes. Pluripotency and undifferentiated stage were confirmed using molecular cell surface markers. Normal karyotype of these cell lines was confirmed. Here we demonstrate that SCNT-h-ESCs differentiate to hematopoietic precursors when cocultured with unmodified, nonirradiated donor BM. We did not use any xenogeneic material for this hematopoietic differentiation. Hematopoietic precursors derived from them expressed cell surface antigens CD45/34. When further cultured with hematopoietic growth factors these hematopoietic precursors formed characteristic myeloid, erythroid, and megakaryocyte lineages. Phenotypic CD34+ cells derived from NT-h-ESCs were functionally similar to their counterparts in primary hematopoietic tissues like BM, umbilical cord, and blood. More terminally differentiated hematopoietic cells derived from h-ESCs under these culture conditions also expressed normal surface antigens like glycophorin A on erythroid cells, CD15 on myeloid cells, and CD41 on megakaryocytes. We report generation of hematopoietic progenitor cells from h-ESC lines by a SCNT technique, with differentiation into further lineages with structural and functional similarities to their adult counterparts in vivo. This novel alternative source of CD34+ stem cells from h-ESC lines generated without any xenogeneic material might be used to create transplantation tolerance, to implement regenerative medicine, and to treat autoimmune disorders.

Embryonic stem cell derived and adult hematopoietic stem cell transplantation for tolerance induction in a renal allograft recipient:--a case report.

UNLABELLED: We generated an human embryonic stem cell (hESC) line to augment chimerism-associated tolerance. A 40-year-old African with chronic glomerulonephritis-chronic renal failure with 100% G6PD enzyme deficiency presented for renal transplantation with a 27-year-old, 6/6 HLA-matched sister as a willing donor. METHOD: We generated an hESC line from the donor's oocytes using long ovarian stimulation protocol simultaneously with tolerance induction protocol. A nuclear transfer (NT)-hESC line was derived by transferring a donor cumulus cell into an enucleated oocyte, subjected to electrical fusion, and cultured for 5 days. ESCs hatched from the blastocyst on day 6 were cocultured with her unmodified bone marrow for 2 days and suspended in Ringer's lactate. Five milliliters of suspension were collected for cell counting, viability, pluripotency, flow cytometry, and karyotyping. The remaining suspension was infused into the periphery of the recipient. Transplantation was performed 1 week later following a negative lymphocytotoxicity cross-match test using no immunosuppression. Peripheral blood chimerism (PBC) was studied using fluorescent in situ hybridization technique. Allograft biopsy was performed on day 7. RESULTS: NT-hESC CD34+ count was 7.6%, viability 100%, karyotyping normal, pluripotency markers: SSEA-1, SSEA-4, OCT-3/4, TRA-1/60:positive; 12% PBC was noted at 1 week after transplantation. Serum creatinine was 1.2 mg%, graft biopsy was unremarkable, and G6PD enzyme deficiency was corrected to 0% at 100 days posttransplant. Liver function tests and hematology profile were unremarkable for graft-versus-host disease. CONCLUSION: This is the first report of tolerance induction using NT-hESC-induced hematopoietic chimerism with synergistic use of adult bone marrow. It was safe and effective.

Iodinated contrast media induced oxidative stress status in patients undergoing urography.

BACKGROUND: The present investigation is an extension of our earlier work on oxidative stress in a number of clinical situations and awareness of potential toxicity of contrast media. It was of interest to understand the relative safety (in terms of balance between oxidants/antioxidants) of the usage of iodinated contrast media in urography. METHODS: Alterations in serum levels of oxidant lipid peroxidation and product malondialdehyde (MDA) and components of anti-oxidant have been studied in 125 patients (with renal calculus disease) to undergo intravenous urographic procedure before and 3 hours following intravenous administration of Trivideo-400 (Iothalmate Sodium 40 ml: 18 gm Iodine) and results of serum levels of MDA (nmol/ml), SOD (mu/ml), Vit-C and Vit-E (mg/dl) were also compared with 135 appropriately age matched controls. RESULTS: The salient features of present investigation indicated definite pattern viz. Significant rise in serum levels of oxidant MDA (4.35 +/- 2.05) and fall in SOD (3.04 +/- 1.04), Vit-C (0.55 +/- 0.17), Vit-E (0.67 +/- 0.20) in patients at pre-IVU stage in comparison with those of controls viz MDA (4.35 +/- 2.05), SOD (3.77 +/- 1.55), Vit-C (0.71 +/- 0.25) and Vit-E (0.80 +/- 0.30). There occurred further elevation in serum levels of MDA (5.28 +/- 2.48) and fall in SOD (3.07 +/- 0.97), Vit-C (0.52 +/- 0.16), Vit-E (0.66 +/- 0.19) showing no significant alterations 3 hours following intravenous IVU procedure. CONCLUSIONS: It is concluded that contrast media induced changes did remain within tolerable physiological limits and no significant changes in anti-oxidant components Vit-E and SOD were suggestive of relative safety of iodinated contrast media usage.

Oxidative stress status in children with nephrotic syndrome.

In continuation of our work on human stress situation and present day awareness of the role of free radical toxicity in a variety of clinical conditions, oxidative stress status (in terms of serum levels of MDA, scavenging enzyme SOD, vitamins: C and E) has been studied in 45 pediatric patients with nephrotic syndrome (further classified as steroid: responders, frequent/ infrequent relapsers, dependents). The results have been compared with 42 appropriately age healthy children as controls. The salient features of the present study centre around typical observations viz significantly increased levels of MDA (7.92 +/- 2.24 nmol/ml), decreased levels of SOD (1.36 +/- 1.01 U/ml), vitamin C (0.49 +/- 0.17 mg/dl) and vitamin E (0.52 +/- 0.19 mg/dl) in children with nephrotic syndrome as a whole when compared with healthy controls [MDA (4.40 +/- 1.31 nmol/ml), SOD (3.04 +/- 1.83 U/ml), vitamin C (0.60 +/- 0.26 mg/dl) and vitamin E (0.68 +/- 0.25 mg/dl) respectively]. An almost similar trend was encountered in different groups as classified. However, maximum fluctuations were observed in steroid dependents. The present observations appear to be suggestive of alternative guidelines to clinicians in the absence of conventional renal biopsy as the procedure. It is felt that children with nephrotic syndrome should regularly take vitamins C and E from the health point of view.

Serum lipoperoxide levels in pregnancy induced hypertension.

With present day awareness of role and toxicity of lipid peroxides, serum levels of lipoperoxides have been investigated in forty pregnancy induced hypertension (PIH) subjects, before and after antihypertensive drug therapy, using full term normotensive patients as control. PIH subjects demonstrated highly significant elevated serum levels of lipoperoxides (mean 3.60 nmol/ml) in comparison (mean 2.15 nmol/ml) with full term normotensive subjects. At post-partum PIH subjects following antihypertensive drug therapy and becoming normotensive showed highly significant fall in serum lipoperoxide (mean level 2.27 nmol/ml). Full term normotensive subjects at post-partum also demonstrated the same pattern. However, observed levels of serum lipoperoxide in PIH subjects at post-partum even after becoming normotensive following treatment were found to be significantly elevated when compared with those of full term normotensives at post-partum (mean level 1.94 nmol/ml). It is felt that monitoring of serum lipoperoxide level can serve as one of the useful parameters in PIH subjects.

Serum lipoperoxide levels in surgical stress of abdominal hysterectomy.

In continuation of our studies on stress situations in female reproductive life span, serum lipid peroxide levels in terms of malondialdehyde (nmol/ml) have been determined in females [(n = 40) (aged between 35-40 years)] to undergo total abdominal hysterectomy (due to dysfunctional uterine bleeding). There occurred a statistically significant though within physiological limits, progressive rise in serum lipid peroxide levels right from preoperative stage 3.51, per operatively 3.56 to a maximum of 4.10 postoperative stage I with a tendency of fall at postoperative stage II 3.94 in comparison with self control on admission 2.97. It is suggestive of the balancing and protective role of scavenging enzyme system like superoxide dismutase also backed by improvement in health status of abdominal hysterectomy patients in terms of ambulation, free intake of food and behavioural positive attitude within 24 hours following operation presumably due to relief from stress of DUB. It is felt from the results of the present investigation that patients undergoing abdominal hysterectomy were free from free radical toxicity.

Effect of renal transplantation (surgical stress) on serum levels of oxidants and reducing system.

BACKGROUND: This study attempts to understand the correlation (if any) between surgical stress of renal transplantation and serum levels of pro-oxidant/antioxidants. METHODS: Twenty-four ESRD patients having undergone transplant surgery followed from day-1 to day-10 postsurgery were investigated for serum levels of MDA, SOD, Vit. C and E. The drug regimen received was immunosuppressant, H2 blockers and antihypertensives as per the situation. RESULTS: The typical observations indicated elevated serum levels of MDA from preoperative stage reaching peak value 24 hrs after surgery followed by a steady fall and achieving minima on the 10th day. As regards antioxidants enzyme SOD, Vit. C, Vit. E were low from pretransplant day reaching minima 24 hours postoperatively and returning to normal from 7th day. CONCLUSIONS: The present investigation has amply shown a typical imbalance between pro-oxidant/antioxidants from pretransplant day up to 24 hrs and there after returning to normal level from 7th day suggestive to desired recovery and surgical stress not a limiting factor in way of health progress renal transplant. Dietary intake of Vit. C and E in mega doses can be a good therapeutic measure.

Oxidative stress status: possible guideline for clinical management of critically ill patients.

BACKGROUND: Critical care medicine has developed in the last few years into a separate scientific discipline and studies related to the outcome after intensive care usually suggest a long hospital stay that becomes cost prohibitive. The majority of problems (death) amongst critically ill patients requiring critical care involve sepsis, inflammation, tissue damage-oxidative stress, oxygen tension PO2, lipid peroxidation. The present investigation involves monitoring of serum levels of MDA, SOD as a possible guideline for severity of clinical situations in critically ill patients. METHODS: Fifty critically ill heterogeneous patients requiring intensive care in the ICU of IKDRC were selected as subjects with ages varying from 17 to 75 years. Serum levels of MDA (ng/ml), SOD (U/ml) were determined right from admission to discharge due to improvement / DAMA / death. MDA and SOD were estimated according to the methods of Buege and Aust et al. (1978), Nandi and Chatterji (1988). RESULTS: Critically ill patients irrespective of the disease process indicated significantly very high serum levels of MDA and low levels of SOD at the time of admission (13.28+/-4.26 ng/ml, 3.80+/-2.60 U/ml, respectively) according to the severity of the prevalent clinical situation. The pattern of serum levels of MDA and SOD according to subsequent clinical performance did indicate a decreasing trend of MDA (oxidant) and fluctuating trend of SOD (antioxidant enzyme except in those who inevitably succumbed to death in spite of adequate clinical management. CONCLUSIONS: The results of the present study have amply revealed the utility and relevance of monitoring oxidative stress in critically ill patients as biochemical markers, cost-effectiveness and role in decision making (withdrawal/continuation of different support modalities) as deemed fit.

Stress and women.

Stress-tension-pain occur more often in health disorders of women than anywhere in the entire spectrum of human illness. No where is the effect of rapid change more apparent than in the changing role of today's women who are subjected to pressures from all sides. Now a days women virtually take part occupy positions, in day to day activities similar to their male counterparts. An outline of conceptual consideration, biochemical overview of stress and intricacies of coping mechanisms are delineated from the point of view of stress situations in female (during reproductive life span) peculiar to them only viz menarche, menstrual cycle, pregnancy-eventualities of pregnancy-labour-delivery, purperium, postpartum psychosis, abortions-induced, spontaneous, menopause. Lipid profile and oxidative stress status as biochemical responses in these stresses were investigated. As expected observed maximal/minimal biochemical responses well correlated with peak time and at termination of stress. Present day emphasis is on menstrual regulation by intake of oral contraceptive to improve health if women prefer not to bleed every month and hormone replacement therapy for elderly women to improve post menopausal quality of life and avoiding menstrual problems at the same time.

Serum lipoperoxides in induced and spontaneous abortions.

Abortion, primarily as a measure of population control, certainly continues to be an emotional, frustrating and stressful event. In continuation of our work on stressful situations in the female life span and biochemical parameters, serum lipid peroxide levels in terms of malondialdehyde (nmol/ml) have been determined in females undergoing abortion [suction curettage (n = 30), Emcredil-induced abortion (n = 30) and spontaneous abortion (n = 40)] and were compared with appropriate gestational controls. Irrespective of the type of abortion, the serum lipid peroxide levels before abortion [mean malondialdehyde concentrations (nmol/ml): suction curettage 2.67, Emcredil-induced abortion 3.22, and spontaneous abortion 3.49] were found to be significantly elevated in comparison with those after abortion (suction curettage 1.91, Emcredil 1.97 and spontaneous abortion 1.95), indicating a maximum at peak time of stress and a minimum at the end of stress. The levels of serum lipid peroxide encountered before abortion were found to be significantly elevated in case of Emcredil-induced abortion and spontaneous abortion when compared with controls (second trimester mean levels 1.82 and first trimester 2.4) whereas the levels before suction curettage were found to be nonsignificant in comparison with controls, indicating a lesser degree of stress. It is felt that monitoring of serum lipid peroxide levels in serum and tissues (placenta), backed by scavenging enzyme superoxide dismutase, can be more helpful for corroborating safety and the risk of free radical toxicity in pregnancy and abortion.

PIP: Abortion, primarily as a measure of population control, continues to be an emotional, frustrating, and stressful event. As an extension of the work the authors have done on stressful situations in the lifespan of the female and its biochemical parameters, serum lipid peroxide levels in terms of malondialdehyde (nmol/mI) have been determined in women undergoing abortion (n=30 for suction curettage), Emcredil-induced abortion (n-30), and spontaneous abortion (n-40) and were compared with appropriate gestational controls. Regardless of the type of abortion, the serum lipid peroxide levels before abortion (mean malondialdehyde concentrations [nmol/ml]: suction curettage 2.67, Emcredil-induced abortion 3.22, and spontaneous abortion 3.49) were significantly elevated in comparison with those after abortion (suction curettage 1.91, Emcredil 1.97, and spontaneous abortion 1.95), indicating a maximum at peak time of stress and a minimum at the end of stress. Levels of serum lipid peroxide encountered prior to abortion were significantly elevated in cases of Emcredil-induced abortion and spontaneous abortion when compared with controls (2nd trimester mean levels 1.82 and 1st trimester 2.4), whereas the levels prior to suction curettage were nonsignificant in comparison with controls. This indicates a lesser degree of stress. It was decided that monitoring of serum lipid peroxide levels in serum and tissues (placenta), backed up by scavenging enzyme superoxide dismutase, can be more helpful in corroborating safety and the risk of free radical toxicity in pregnancy and abortion.