Chronic stress, sense of coherence and risk of type 2 diabetes mellitus.

AIMS: The study was conducted to ascertain whether chronic stress and sense of coherence are associated with risk of type 2 diabetes mellitus. METHODS: Stress questionnaires - Presumptive Stressful Life Events Scale (PSLES), Perceived Stress Scale (PSS) and Sense of Coherence (SOC) - were administered to 500 Newly Detected Diabetes Mellitus (NDDM) cases and 500 Normal Glucose Tolerance (NGT) controls recruited following 75 g OGTT. Assessment of stress was completed before the diagnosis of diabetes was revealed to them. RESULTS: PSLES and PSS scores were significantly higher and SOC score was significantly lower in NDDM subjects compared to those with NGT. PSLES and PSS correlated positively with anthropometric parameters (waist circumference, BMI), glycemic parameters (FPG, 2 hPG, A1C) and HOMA-IR and inversely with HOMA-ß whereas SOC correlated inversely with glycemic parameters (FPG, 2 hPG, A1C) and HOMA-IR and positively with HOMA-ß. In stepwise logistic regression analysis, SOC emerged as the strongest independent predictor of diabetes (OR: 0.774) after HOMA-IR (OR: 1.621) and BMI (OR: 1.288). Other significant predictors included PSS (OR:1.153), PSLES-LT (OR: 1.005) and HOMA-ß (OR: 0.894). CONCLUSION: Chronic stress and low sense of coherence are associated with a higher risk of type 2 diabetes mellitus.

Role of Antibiotic Cement Coated Nailing in Infected Nonunion of Tibia.

INTRODUCTION: Infected nonunion of long bones is a chronic and debilitating disorder. It is more difficult to deal with when the implant used for internal fixation itself becomes a potential media for infection because of bacterial adhesion and biofilm formation. Traditionally, it is managed by two-stage procedure for controlling the infection first and then treating the nonunion. This study has been undertaken to explore antibiotic cement coated nailing as single stage treatment modality for treating infection and achieving stability at the same time. MATERIALS AND METHODS: Twenty patients (above 18 years of age) with infected nonunion of tibia with bone gap less than 2 cm were managed using antibiotic cement coated K-nail. Antibiotic cement nail was prepared using endotracheal tube method. Antibiotics used were a combination of vancomycin and teicoplanin. RESULTS: Infection was controlled in 95% of the patients. Bony union was achieved in 12 of 20 (60%) patients with antibiotic cement nailing as the only procedure with average time of union of 32 weeks. Remaining 8 patients required additional procedures like bone grafting or exchange nailing and these were done in six patients, with union of fracture. Two patients refused to undergo further procedures. Complications encountered were difficult nail removal in three cases, broken nail in two cases, and bent nail in one case. Recurrence of infection was observed in two patients. Average period of follow-up was 13 months. CONCLUSION: Antibiotic cement impregnated nailing is a simple, economical and effective single stage procedure for the management of infected nonunion of tibia. It is advantageous over external fixators, as it eliminates the complications of external fixators and has good patient compliance. The method utilizes existing easily available instrumentation and materials and is technically less demanding, and therefore can be performed at any general orthopaedic center.

Primary Mucoepidermoid Carcinoma of the Lacrimal Sac -  a Case Report and Literature Review.

BACKGROUND: Lacrimal sac tumors are very rare and are often missed because patients present with features consistent with chronic dacryocystitis. Squamous cell carcinoma is the common-est lacrimal sac malignancy. Although primary mucoepidermoid carcinomas of the lacrimal sac are rare, they are locally aggressive. Furthermore, their proximity to vital structures and the skull base makes them potentially life-threatening. Multidisciplinary management is required, and wide excision followed by chemoradiation is the recommended treatment. CASE: Here, we report a 65-year-old male who presented with watering eyes and a mass in the region of the medial canthus. A dia-gnosis of primary mucoepidermoid carcinoma of the lacrimal sac was made, and the case was managed successfully with radical surgery and reconstruction. The tumor was resected using the extended Lynch-Howarth incision and the resulting defect was reconstructed using a forehead flap. Histopathological examination of the excised specimen revealed mucoepidermoid carcinoma. Immunohistochemical analysis revealed that the speci-men was positive for epithelial growth factor receptor and Ki-67 protein. The patient was referred for post-operative chemoradiation. The literature is reviewed and pathological features, including immunohistochemistry are discussed. CONCLUSION: Primary mucoepidermoid carcinoma of the lacrimal sac is a rare, locally aggressive tumor that is often mistaken for dacryocystitis. The treatment of choice is radical surgery followed by chemoradiation. KEY WORDS: lacrimal sac -  mucoepidermoid carcinoma -  epithelial growth factor receptor -  Ki-67 protein.

Wrist fusion through centralisation of the ulna for recurrent giant cell tumour of the distal radius.

PURPOSE: To review the outcome of wide resection and wrist fusion through centralisation of the ulna for recurrent giant cell tumour (GCT) of the distal radius in 10 patients. METHODS: Records of 7 women and 3 men aged 25 to 40 years who underwent wide resection and wrist fusion through centralisation of the ulna for grade I (n=6) and grade II (n=4) recurrent GCT of the distal radius were reviewed. Functional outcome was evaluated using the modified Musculoskeletal Tumor Society scoring system. Grip strength and range of motion of the metacarpophalangeal joint were also assessed. RESULTS: The mean follow-up period was 45 (range, 30-60) months. All tumours were benign and there was no evidence of metastasis. The mean grip strength was 45% (range, 30-70%) of the contralateral side. All patients achieved normal range of motion of the metacarpophalangeal and interphalangeal joints, but 4 patients had an extensor lag of the thumb. The mean time to wrist fusion was 6 (range, 4-8) months: 3 to 4 months for the 6 patients with bone grafting and 5 to 6 months for the 4 patients without bone grafting who developed painful subluxation necessitating total wrist arthrodesis using a plate and morcellised bone grafts. CONCLUSION: Wrist fusion through centralisation of the ulna for recurrent GCT of the distal radius is a viable option, despite a loss of wrist motion.

Modified approach to powered endoscopic dacryocystorhinostomy.

OBJECTIVE: This paper presents modifications to powered endoscopic dacryocystorhinostomy without stenting for the management of acquired nasolacrimal duct obstruction. STUDY DESIGN: A retrospective, non-randomised review. METHOD: A total of 105 patients with epiphora secondary to acquired nasolacrimal duct obstruction were operated on by our modified powered endoscopic dacryocystorhinostomy approach. RESULTS: Of 105 patients, 100 (95.2 per cent) have remained asymptomatic. One patient with a narrow ostium had improved symptoms, while four patients continued to experience persistent symptoms (treatment failures). CONCLUSION: The modified powered endoscopic dacryocystorhinostomy approach led to excellent results, without bone exposure and granulations, and with early mucosalisation.

Assessment of Oxidative Stress in Patients with Premature Canities.

CONTEXT: Premature canities is a common, yet unexplored disorder. Oxidative stress levels have been evaluated within the greying hair follicle but not in the sera of patients with premature canities. AIMS: To evaluate the oxidative stress parameters in the sera of patients with premature canities. SETTINGS AND DESIGN: A pilot case-controlled study, conducted in a tertiary care setup in Delhi during November 2011 to December 2012. MATERIALS AND METHODS: Fifty-two self-reporting cases of premature canities (age of onset <20 years) and 30 healthy controls were recruited from outpatient Department of Dermatology. Oxidative stress parameters (serum malonaldehyde (MDA), whole blood reduced glutathione (rGSH) and serum ferric reducing antioxidant potential [FRAP]) were assessed in cases and controls. Mann-Whitney test was used to compare the oxidative stress parameters between the two groups (SPSS version 17.0, SPSS Inc, Chicago, USA; P < 0.05 considered as significant). RESULTS: The age and sex distribution of cases and controls was comparable. The mean serum levels of MDA were higher in cases than controls (3.7 ± 1.6 nmol/ml vs. 2.8 ± 1.5 nmol/ml; P = 0.01). The GSH levels were lower in the cases than controls (31.5 ± 8.9 mg/dl vs. 36.6 ± 16.9 mg/dl; P = 0.064). Similarly, the mean FRAP levels were lower in the cases than controls (400 ± 70 nmol/ml vs. 430 ± 80 nmol/ml; P = 0.038). CONCLUSIONS: Patients with premature canities had a higher level of pro-oxidants and lower levels of antioxidants than controls. This is the first humble attempt to document the oxidative stress parameters in sera of patients with premature canities, further studies with larger sample size are required to reach a definite conclusion.

Thalamic superoxide and peroxide handling capacity (SPHC): An experimental study with aluminum, ethanol and tocopherol in rats.

Superoxide and peroxide handling capacity (SPHC) is an important determinant of oxidative stress. Neurotoxic impacts of aluminum are associated with oxidant imbalance. Here, we studied the influence of aluminum on oxidative stress parameters, antioxidative enzymes and SPHC of thalamic area on pro-oxidant (ethanol) and antioxidant (α-tocopherol) exposure. Two sets of male Wistar rats were divided into 8 groups (6 each) and exposed to aluminum (10 mg/Kg body wt.), ethanol (0.6 g/Kg body wt.) and α-tocopherol (5 IU/day) for 4 wk, each having respective control group. Levels of reduced glutathione (GSH), lipid peroxidation (TBARS) along with activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) of thalamic area were estimated for each group. Glutathione-independent superoxide peroxide handling capacity (GI-SPHC) and glutathione-dependent superoxide peroxide handling capacity (GD-SPHC) were calculated from the GPx, CAT and SOD values. Concomitant exposure to aluminum and ethanol demonstrated significant increase in SOD activity and significant decrease in GPx activity compared to the control group, while lone aluminum-exposed rats showed raised GR activity, without alterations in GPx and SOD activities. However, significant reduction of both GI- and GD- SPHC were found in ethanol-exposed groups. α-Tocopherol supplementation could resist most of the alterations. In addition, current antioxidant exposure reduced the inherent GD-SPHC, and thus, made thalamic area more vulnerable to oxidant threat. The present study corroborates the thalamic susceptibility to aluminum-augmented oxidant imbalance and suggests cautious use of antioxidant supplementation against neurodegenerative disorders.

Endocrine stress responses and risk of type 2 diabetes mellitus.

This study was carried to ascertain whether stress responses are associated with abnormalities in glucose tolerance, insulin sensitivity and pancreatic beta cell function and risk of type 2 Diabetes Mellitus. Salivary cortisol, a marker of hypothalamic-pituitary-adrenal (HPA) axis and salivary α-amylase, a marker of sympathetic nervous system (SNS) were compared in 125 subjects of newly detected diabetes mellitus (NDDM) and normal glucose tolerance (NGT) subjects who were diagnosed on the basis of oral glucose tolerance test (OGTT). Assessment of stress in them was done through stress scales - presumptive stressful life events scale (PSLES), perceived stress scale (PSS) and sense of coherence (SOC) and correlated with these and other stress response markers. Significantly higher 10 pm salivary cortisol and post dexamethasone salivary cortisol were found in NDDM subjects as compared to NGT. 10 pm salivary cortisol correlated significantly with fasting plasma glucose (FPG), 2 h plasma glucose (2h PG) and glycated hemoglobin (HbA1c) while post dex salivary cortisol correlated with 2h PG, HbA1c and salivary α-amylase with 2h PG. Stepwise logistic regression analysis showed that body mass index (OR: 1.840), SOC (OR: 0.688) and 10 pm salivary cortisol (OR: 1.427) were the strongest predictors of NDDM. The results of the present study indicate that NDDM subjects display significantly higher chronic stress and stress responses when compared to subjects with NGT. Chronic stress and endocrine stress responses are significantly associated with glucose intolerance, insulin resistance and diabetes mellitus.

Endocrine stress responses and risk of type 2 diabetes mellitus.

This study was carried to ascertain whether stress responses are associated with abnormalities in glucose tolerance, insulin sensitivity and pancreatic beta cell function and risk of type 2 Diabetes Mellitus. Salivary cortisol, a marker of hypothalamic-pituitary-adrenal (HPA) axis and salivary α-amylase, a marker of sympathetic nervous system (SNS) were compared in 125 subjects of newly detected diabetes mellitus (NDDM) and normal glucose tolerance (NGT) subjects who were diagnosed on the basis of oral glucose tolerance test (OGTT). Assessment of stress in them was done through stress scales - presumptive stressful life events scale (PSLES), perceived stress scale (PSS) and sense of coherence (SOC) and correlated with these and other stress response markers. Significantly higher 10 pm salivary cortisol and post dexamethasone salivary cortisol were found in NDDM subjects as compared to NGT. 10 pm salivary cortisol correlated significantly with fasting plasma glucose (FPG), 2 h plasma glucose (2h PG) and glycated hemoglobin (HbA1c) while post dex salivary cortisol correlated with 2h PG, HbA1c and salivary α-amylase with 2h PG. Stepwise logistic regression analysis showed that body mass index (OR: 1.840), SOC (OR: 0.688) and 10 pm salivary cortisol (OR: 1.427) were the strongest predictors of NDDM. The results of the present study indicate that NDDM subjects display significantly higher chronic stress and stress responses when compared to subjects with NGT. Chronic stress and endocrine stress responses are significantly associated with glucose intolerance, insulin resistance and diabetes mellitus.

Kruppel-like factor 4 signals through microRNA-206 to promote tumor initiation and cell survival.

Tumor cell heterogeneity poses a major hurdle in the treatment of cancer. Mammary cancer stem-like cells (MaCSCs), or tumor-initiating cells, are highly tumorigenic sub-populations that have the potential to self-renew and to differentiate. These cells are clinically important, as they display therapeutic resistance and may contribute to treatment failure and recurrence, but the signaling axes relevant to the tumorigenic phenotype are poorly defined. The zinc-finger transcription factor Kruppel-like factor 4 (KLF4) is a pluripotency mediator that is enriched in MaCSCs. KLF4 promotes RAS-extracellular signal-regulated kinase pathway activity and tumor cell survival in triple-negative breast cancer (TNBC) cells. In this study, we found that both KLF4 and a downstream effector, microRNA-206 (miR-206), are selectively enriched in the MaCSC fractions of cultured human TNBC cell lines, as well as in the aldehyde dehydrogenase-high MaCSC sub-population of cells derived from xenografted human mammary carcinomas. The suppression of endogenous KLF4 or miR-206 activities abrogated cell survival and in vivo tumor initiation, despite having only subtle effects on MaCSC abundance. Using a combinatorial approach that included in silico as well as loss- and gain-of-function in vitro assays, we identified miR-206-mediated repression of the pro-apoptotic molecules programmed cell death 4 (PDCD4) and connexin 43 (CX43/GJA1). Depletion of either of these two miR-206-regulated transcripts promoted resistance to anoikis, a prominent feature of CSCs, but did not consistently alter MaCSC abundance. Consistent with increased levels of miR-206 in MaCSCs, the expression of both PDCD4 and CX43 was suppressed in these cells relative to control cells. These results identify miR-206 as an effector of KLF4-mediated prosurvival signaling in MaCSCs through repression of PDCD4 and CX43. Consequently, our study suggests that a pluripotency factor exerts prosurvival signaling in MaCSCs, and that antagonism of KLF4-miR-206 signaling may selectively target the MaCSC niche in TNBC.

Drug development from natural resource: a systematic approach.

Modern research in drug discovery from medicinal plants involves a multidimensional approach combining botanical, phytochemical, biochemical combinatorial chemistry and bioassay-guided fractionation approaches. Natural sources continue to provide an alternative as pharmacological leads against various devastating diseases such as diabetes, CVD, cancer etc. Nowadays, there is enormous requirement of safe and effective drugs in the world. This has prompted scientists to revert back towards natural resources as a potential source of therapeutics for treatment and management of such chronic and fatal diseases. However, there are certain serious challenges and limitations in this field including scale up and commercialization of active compounds which allow only one in thousand lead molecules to be developed as drug. A systematic and scientific approach is an essential requirement for drug development from natural resource. This mini review provides an overview of the methods involved in natural product research starting from crude plant extract to bioactive pharmacological lead. Moreover, it also discusses the limitations of working concerning the bioactivity of medicinal plants.

Regulation of anti-apoptotic signaling by Kruppel-like factors 4 and 5 mediates lapatinib resistance in breast cancer.

The Kruppel-like transcription factors (KLFs) 4 and 5 (KLF4/5) are coexpressed in mouse embryonic stem cells, where they function redundantly to maintain pluripotency. In mammary carcinoma, KLF4/5 can each impact the malignant phenotype, but potential linkages to drug resistance remain unclear. In primary human breast cancers, we observed a positive correlation between KLF4/5 transcript abundance, particularly in the human epidermal growth factor receptor 2 (HER2)-enriched subtype. Furthermore, KLF4/5 protein was rapidly upregulated in human breast cancer cells following treatment with the HER2/epidermal growth factor receptor inhibitor, lapatinib. In addition, we observed a positive correlation between these factors in the primary tumors of genetically engineered mouse models (GEMMs). In particular, the levels of both factors were enriched in the basal-like tumors of the C3(1) TAg (SV40 large T antigen transgenic mice under control of the C3(1)/prostatein promoter) GEMM. Using tumor cells derived from this model as well as human breast cancer cells, suppression of KLF4 and/or KLF5 sensitized HER2-overexpressing cells to lapatinib. Indicating cooperativity, greater effects were observed when both genes were depleted. KLF4/5-deficient cells had reduced basal mRNA and protein levels of the anti-apoptotic factors myeloid cell leukemia 1 (MCL1) and B-cell lymphoma-extra large (BCL-XL). Moreover, MCL1 was upregulated by lapatinib in a KLF4/5-dependent manner, and enforced expression of MCL1 in KLF4/5-deficient cells restored drug resistance. In addition, combined suppression of KLF4/5 in cultured tumor cells additively inhibited anchorage-independent growth, resistance to anoikis and tumor formation in immunocompromised mice. Consistent with their cooperative role in drug resistance and other malignant properties, KLF4/5 levels selectively stratified human HER2-enriched breast cancer by distant metastasis-free survival. These results identify KLF4 and KLF5 as cooperating protumorigenic factors and critical participants in resistance to lapatinib, furthering the rationale for combining anti-MCL1/BCL-XL inhibitors with conventional HER2-targeted therapies.

Does type of feeding in infancy influence lipid profile in later life?

OBJECTIVE: To compare the lipid profile of exclusively breastfed and mixed-fed, term, appropriate for gestation age infants from 6 mo to 1 y of age. METHODS: This prospective comparative study included one hundred ninety nine consecutive term healthy infants; 105 on exclusive breastfeeding (EBF) and 94 receiving mixed feeding (MF). These children were recruited at 6 mo of age and followed till 1 y of age. Serum lipid levels of babies were determined at recruitment (6 mo), 9 mo and 1 y of age. Statistical analysis was carried out using SPSS software. RESULTS: The mean total cholesterol (TC) at 6 mo in exclusively breastfed group (156.38 ± 50.42 mg/dl) was significantly higher than mixed fed group (139.5 ± 37.59 mg/dl). At 9 mo, high density lipoprotein cholesterol (HDL-C) and triglycerides (TG) levels were significantly different in EBF group than MF group. The lipid profile of both group of babies was comparable at 1 y of age. The HDL-C: LDL-C ratio was significantly different between the two groups (higher in breastfed group) at 1 y. CONCLUSIONS: The present study highlights differential lipid profile of exclusively breastfed infants and mixed fed infants.

Protective effects of Eugenia jambolana extract versus N-acetyl cysteine against cisplatin-induced damage in rat testis.

To assess the protective effects of Eugenia jambolana extract (EJE) or N-acetyl cysteine (NAC) on testis, cisplatin (CIS, 5 mg kg(-1) bw, single dose) was administered either alone or along with EJE (25 mg kg(-1) bw, alternate day) or NAC (150 mg kg(-1) bw, Day 1 and 4) for 7 days. Significant alterations in serum LH, FSH and testosterone were observed in CIS group which were effectively modulated by EJE or NAC supplementation. Upregulation of 3ß-HSD gene indicated the rise in functional Leydig cells. This was further confirmed from the identical improvement in hCG-stimulated testosterone production in isolated Leydig cells. Reduction in oxidative stress was associated with restoration of total antioxidant capacity and glutathione levels, and activation of antioxidant enzymes, SOD, catalase, glutathione s-transferase (GST) and glutathione reductase (GR). CIS-induced apoptosis of germ and Leydig cells was contained by both NAC and EJE intervention by effective modulation of apoptotic markers in the extrinsic, intrinsic and other pathways of metazoan apoptosis. Taken together, the study findings establish the potential of EJE as a therapeutically better antioxidant than NAC for use in curtailing the adverse effects of anticancer drugs on testicular function.

Waardenburg syndrome presenting with constipation since birth.

BACKGROUND: Shah-Waardenburg syndrome is Waardenburg syndrome associated with Hirschsprung's disease. CASE CHARACTERISTICS: A 10-day-old full-term male neonate of Waardenburg syndrome presented with constipation since birth along with features of small bowel obstruction. OBSERVATION: Exploratory laparotomy revealed distended proximal jejunal and ileal loops along with microcolon; an ileostomy was performed. OUTCOME: Postoperatively patient developed sepsis and died. Histopathology confirmed total colonic aganglionosis. MESSAGE: Suspect familial Shah-Waardenburg syndrome in a neonate of Waardenburg syndrome presenting with constipation since birth or intestinal obstruction.

Aluminum and ethanol induce alterations in superoxide and peroxide handling capacity (SPHC) in frontal and temporal cortex.

Aluminum is an omnipresent neurotoxicant and has been associated with several neuropathological disorders. Cerebrum and cerebellum have been shown to face augmented oxidative stress when animals are exposed to aluminum and high doses of ethanol. To establish the link between oxidative stress and neurobehavioral alterations, the present study was conducted to determine the extent of oxidative stress in low levels of pro-oxidant (ethanol exposure) status of the functionally discrete regions of the cerebrum. Male Wistar rats were exposed to aluminum (10 mg/kg body wt) and ethanol (0.2-0.6 g/kg body wt) for 4 weeks. Spontaneous motor activity (SMA) and Rota-Rod performances (RRP) were recorded weekly during the period of exposure. At the end of 4th week, oxidative stress parameters were determined from the homogenized cerebral tissue. GSH-independent superoxide peroxide handling capacity (GI-SPHC) and GSH-dependent superoxide peroxide handling capacity (GD-SPHC) were determined for FC and TC upon exposure to ethanol in the absence and presence of aluminum exposure. Aluminum was found to augment the oxidative stress at higher doses (0.6 g Ethanol/kg body wt) of ethanol, particularly in FC. The SPHC of FC was also found to be compromised significantly in aluminum-ethanol co-exposed animals. It was concluded that even though the manifestation of oxidative stress was not observed as revealed by assaying the widely used oxidative stress biochemical markers (indices), aluminum and ethanol (low doses) exposure induced alterations in the handling capacity of oxidant imbalance that could be recognized by studying the SPHC of FC. Comparison of GD-SPHC and GI-SPHC offered a possible mechanism of compromised SPHC in FC. This observation is likely to offer insights into the mechanism of association between aluminium exposure and behavioral changes in neurodegenerative disorders towards therapeutic strategies for these disorders.

Biomarkers for early detection of risk in female patients with coronary artery disease: pilot study.

In women with coronary artery disease (CAD), clinical presentation is different enough from men which leads to missed or delayed diagnosis. Biomarkers can be used for assessment of CAD patients. In case control study, we analyzed blood samples of 30 controls, 30 cases of Unstable Angina (UA) and 30 cases of Myocardial Infarction (MI) for Pro-inflammatory markers (hs-CRP, IL-6, ICAM-1) and Pregnancy Associated Plasma Protein-A (PAPP-A). Based on discriminant analysis, hs-CRP is the potential marker to discriminate cases of UA from controls while PAPP-A is the reliable marker which can discriminate the cases of MI from UA and controls.

Cytoprotective effects of fruit pulp of Eugenia jambolana on H 2 O 2 -induced oxidative stress and apoptosis in rat Leydig cells in vitro.

This study was undertaken to investigate the cytoprotective effect of the fruit pulp of Eugenia jambolana (50-250 µg ml(-1) ) against the damage induced by H 2 O 2 (100 µm) exposure to Leydig cells in vitro. Cell survival with extract was found comparable to similar effects by N-acetyl-l-cysteine. H 2 O 2 -induced rise in thiobarbituric acid reactive substance formation and decline in the activity and expression of antioxidant enzymes like superoxide dismutase, catalase and glutathione-s-transferase were effectively checked. Cellular glutathione and total antioxidant capacity demonstrated significant improvement. The increase in expression of inducible nitric oxide (NO) synthase leading to NO production was successfully countered. Co-treatment of the extract helped in the down-regulation of caspase-3 and poly-ADP-ribose polymerase resulting in a significant reduction in Leydig cell apoptosis induced by H 2 O 2 . Upstream marker proteins of extrinsic (caspase-8, Fas, FasL) and intrinsic (caspase-9) pathway of metazoan apoptosis were identically down-regulated. The Bcl-2 family of proteins, though, remained unaffected. The extract also positively modulated the other marker proteins like c-Jun NH 2 -terminal kinase, p38, Akt, nuclear factor-κB, c-Fos, cellular FLICE-inhibitory protein, cyclooxygenase-2 and p53. Taken together, the above-mentioned findings establish the anti-oxidative and anti-apoptotic potency of the extract that ameliorates the H 2 O 2 -induced adverse effects on rat Leydig cells in vitro.

siRNA as a tool to delineate pathway channelization in H(2)O(2) induced apoptosis of primary Leydig cells in vitro.

Using siRNA as a tool, the channelization of pathway in H(2)O(2) induced apoptosis of primary Leydig cells was investigated in vitro. Exposure (4 h) to H(2)O(2) (250 µM) induced maximum apoptosis but affected Leydig cell viability significantly. Therefore, expression of apoptotic marker genes, caspase-8, -9, -3 and polyadenosine ribose polymerase was subsequently investigated using the same concentration post 1 h exposure. Incubation with siRNA (20 nM) either for caspase-8 or -9, inhibited their individual expressions by 55-60 % and activity, 50-55 %. The inhibition efficiency using siRNA was comparable with post- or pre-H(2)O(2) treatment of cells. Like siRNA, Eugenia jambolana (100 µg/ml) plant extract too, effectively countered over-expression of all apoptotic marker proteins. Silencing expressions of caspase 8 but not 9 through siRNA leads to a profound inhibition of caspase 3 implying that H(2)O(2) induced Leydig cell apoptosis is preferably channeled through extrinsic and later extending to other pathways.