Signaling pathways and their potential therapeutic utility in esophageal squamous cell carcinoma

Esophageal cancer is a complex gastrointestinal malignancy with an extremely poor outcome. Approximately 80% of cases of this malignancy in Asian countries including India are of squamous cell origin, termed Esophageal Squamous Cell Carcinoma (ESCC).The five-year survival rate in ESCC patients is less than 20%. Neo-adjuvant chemo-radiotherapy (NACRT) followed by surgical resection remains the major therapeutic strategy for patients with operable ESCC. However, resistance to NACRT and local recurrence after initial treatment are the leading cause of dismal outcomes in these patients. Therefore, an alternative strategy to promote response to the therapy and reduce the post-operative disease recurrence is highly needed. At the molecular level, wide variations have been observed in tumor characteristics among different populations, nevertheless, several common molecular features have been identified which orchestrate disease progression and clinical outcome in the malignancy. Therefore, determination of candidate molecular pathways for targeted therapy remains the mainstream idea of focus in ESCC research. In this review, we have discussed the key signaling pathways associated with ESCC, i.e., Notch, Wnt, and Nrf2 pathways, and their crosstalk during disease progression. We further discuss the recent developments of novel agents to target these pathways in the context of targeted cancer therapy. In-depth research of the signaling pathways, gene signatures, and a combinatorial approach may help in discovering targeted therapy for ESCC.

Signaling pathways and their potential therapeutic utility in esophageal squamous cell carcinoma.

Esophageal cancer is a complex gastrointestinal malignancy with an extremely poor outcome. Approximately 80% of cases of this malignancy in Asian countries including India are of squamous cell origin, termed Esophageal Squamous Cell Carcinoma (ESCC).The five-year survival rate in ESCC patients is less than 20%. Neo-adjuvant chemo-radiotherapy (NACRT) followed by surgical resection remains the major therapeutic strategy for patients with operable ESCC. However, resistance to NACRT and local recurrence after initial treatment are the leading cause of dismal outcomes in these patients. Therefore, an alternative strategy to promote response to the therapy and reduce the post-operative disease recurrence is highly needed. At the molecular level, wide variations have been observed in tumor characteristics among different populations, nevertheless, several common molecular features have been identified which orchestrate disease progression and clinical outcome in the malignancy. Therefore, determination of candidate molecular pathways for targeted therapy remains the mainstream idea of focus in ESCC research. In this review, we have discussed the key signaling pathways associated with ESCC, i.e., Notch, Wnt, and Nrf2 pathways, and their crosstalk during disease progression. We further discuss the recent developments of novel agents to target these pathways in the context of targeted cancer therapy. In-depth research of the signaling pathways, gene signatures, and a combinatorial approach may help in discovering targeted therapy for ESCC.

Structural equation modelling analysis determining causal role among methyltransferases, methylation, and apoptosis during human pregnancy and abortion.

The human implantation failure during first trimester leads to spontaneous abortions. Spontaneous abortions are consecutive and occur twice or thrice (with or without prior live births) due to factors which are either maternal or fetal. However, it also constitutes of unknown etiology; known as unexplained recurrent spontaneous abortions (URSA). In this study, the medical terminated human normal early pregnancies (NEP) of the first trimester were taken as control samples, the normal decidual sample whose molecular and epigenetic changes were compared with that of decidua of human URSA subjects. Apoptosis-related genes reported in consecutive recurrent pregnancy loss became the basis for this study. So, in this study, we evaluated the hypothesis that "p53 methylation level through methyltransferases (G9aMT and DNMT1) implicates the fate of embryo towards sustenance or cessation of pregnancy". Further, the interaction between P53, BAX, BCL-2, CASPASE-6, G9aMT, DNMT-1, and methylated p53 expression level(s) during the first trimester of both URSA and NEP are included in this study. The degree of p53 methylation during the first trimester is found to be significant and positively correlated with that of G9aMT (p < 0.05), BCL-2 (p < 0.001), and DNMT1 (p < 0.001) at both transcript and protein level. A significant and negative correlation (with p-value < 0.001) between the degree of p53 methylation during the first trimester and that of the expression level of TUNEL assay (Apoptosis), P53, BAX, and CASPASE-6 are also observed in the present study. A positive correlation between apoptosis and a higher level of p53 expression (which is possibly due to low degree of p53 methylation) is observed both at the transcript and protein level in URSA which is in line with our findings. The analysis performed using structural equation modelling (SEM) further throws light on the causal relationship between sustenance of pregnancy or URSA during the first trimester of a human pregnancy and degree of methylation of p53 which is closely correlated with the interaction between G9aMT, DNMT1, BCL-2, BAX, P53, CASPASE-6, and apoptosis.

Comparison of a grain-based diet supplemented with synthetic vitamin E versus a lucerne (alfalfa) hay-based diet fed to lambs in terms of carcass traits, muscle vitamin E, fatty acid content, lipid oxidation, and retail colour of meat.

Dietary supplementation of vitamin E (VitE) in a synthetic or natural form was examined. Forty-eight lambs were assigned (n = 16) to either a grain-based diet at moderate (MOD, 42 mg∙kg-1 VitE as all-rac α-tocopheryl acetate) or supranutritional (SUP, 285 mg∙kg-1 of vitE) levels of synthetic VitE or a lucerne hay-based diet (LUC; 37 mg∙kg-1 VitE) for 8 weeks. Meat from the LUC group had lower muscle n-6 and PUFA levels compared to meat from the MOD and SUP groups. Despite a similar VitE intake, muscle VitE was higher for LUC compared to MOD, while SUP lambs showed the highest VitE. Lipid oxidation did not differ between groups. For fresh meat, redness tended to be higher in LUC fed lambs than the other two groups, but brownness formation was only lower than the SUP group. For aged meat colour stability, redness tended to be higher in lambs fed SUP and LUC, whereas highest browning occurred in the MOD group.

Effects of a short-term supranutritional selenium supplementation on redox balance, physiology and insulin-related metabolism in heat-stressed pigs.

Heat stress (HS) disrupts redox balance and insulin-related metabolism. Supplementation with supranutritional amounts of selenium (Se) may enhance glutathione peroxidase (GPX) activity and reduce oxidative stress, but may trigger insulin resistance. Therefore, the aim of this experiment was to investigate the effects of a short-term high Se supplementation on physiology, oxidative stress and insulin-related metabolism in heat-stressed pigs. Twenty-four gilts were fed either a control (0.20 ppm Se) or a high Se (1.0 ppm Se yeast, HiSe) diet for 2 weeks. Pigs were then housed in thermoneutral (20°C) or HS (35°C) conditions for 8 days. Blood samples were collected to study blood Se and oxidative stress markers. An oral glucose tolerance test (OGTT) was conducted on day 8 of thermal exposure. The HS conditions increased rectal temperature and respiration rate (both p < .001). The HiSe diet increased blood Se by 12% (p < .05) and ameliorated the increase in rectal temperature (p < .05). Heat stress increased oxidative stress as evidenced by a 48% increase in plasma advanced oxidized protein products (AOPPs; p < .05), which may be associated with the reductions in plasma biological antioxidant potential (BAP) and erythrocyte GPX activity (both p < .05). The HiSe diet did not alleviate the reduction in plasma BAP or increase in AOPPs observed during HS, although it tended to increase erythrocyte GPX activity by 13% (p = .068). Without affecting insulin, HS attenuated lipid mobilization, as evidenced by a lower fasting NEFA concentration (p < .05), which was not mitigated by the HiSe diet. The HiSe diet increased insulin AUC, suggesting it potentiated insulin resistance, although this only occurred under TN conditions (p = .066). In summary, HS induced oxidative stress and attenuated lipid mobilization in pigs. The short-term supranutritional Se supplementation alleviated hyperthermia, but did not protect against oxidative stress in heat-stressed pigs.

Is VEGF under-expressed in Indian children with Perthes disease?

BACKGROUND: The role of vascular endothelial growth factor (VEGF) after ischaemic necrosis of the femoral head in Legg-Calve-Perthes disease (LCPD) has not been adequately studied in humans, especially in Indian population. Therefore, we aimed to evaluate the serum levels of VEGF-A in Indian children with various stages of LCPD and compare them with those of an age- and sex-matched control group of healthy children. METHODS: In this case-control study, we enrolled 42 children (below 14 years age) suffering from LCPD and 21 age- and sex-matched healthy controls. Patients were classified radiographically according to Waldenstrom's classification. Serum VEGF-A was estimated by sandwich enzyme-linked immunosorbent assay technique. The serum values were compared between the patient group and the control group, as well as between the Waldenstrom subgroups. Results were expressed as means with ranges or median with interquartile range. RESULTS: The mean age in the patient as well as the control group was 9 years (range 4-13 years). The median value (interquartile range) of serum VEGF-A was 162.5 pg/ml (673.75 pg/ml) in the patient group and 652 pg/ml (190.5 pg/ml) in the control group (p = 0.013). When compared between lower Waldenstrom stages (initial stage + stage of fragmentation) and higher Waldenstrom stages (re-ossification stage + stage of healing), the mean values of serum VEGF-A were 464.7 pg/ml (range 0-2211 pg/ml) and 301.1 pg/ml (range 0-1910 pg/ml), respectively (p = 0.305). CONCLUSIONS: VEGF is under-expressed in Indian children suffering from LCPD. As VEGF acts as a key regulator of endochondral ossification, our finding may open new therapeutic approaches to the disease. Also, serum VEGF may act as a valuable marker for the follow-up of the disease. Our study also provides baseline data about serum VEGF-A levels in Indian cohort of LCPD patients. Future multi-centre studies are warranted with a larger sample size to fully appreciate the patho-physiological changes in VEGF occurring in LCPD.

Effects of chromium supplementation on physiology, feed intake, and insulin related metabolism in growing pigs subjected to heat stress.

Improving insulin sensitivity may reduce impacts of heat stress (HS) in pigs by facilitating heat dissipation. Chromium (Cr) has been reported to improve insulin sensitivity in pigs. Therefore, the aim of this experiment was to investigate whether Cr supplementation can mitigate HS in growing pigs. Thirty-six gilts were randomly assigned to 2 diets containing 0 (control) or 400 ppb Cr. After 14 d the supplemented pigs were allocated to either 8 d thermoneutral (20°C constant; TN) or cyclic HS (35°C, 0900 h to 1700 h) conditions and continued their respective diet (n = 9 per group). Growth performance was recorded during the 14-d supplementation period. The physiological responses to HS were monitored by measuring respiration rate, rectal temperature, blood gas chemistry, and feed intake during thermal exposure. Kinetics of plasma glucose, insulin and NEFA were studied by intravenous glucose tolerance test (IVGTT) on d 8 of thermal treatment. Results showed Cr alleviated the HS-increased rectal temperature (P < 0.05) and respiration rate (P < 0.01) at 1300 h and 1600 h during thermal exposure. However, Cr did not mitigate the reduction in average daily feed intake which was reduced by 35% during HS or the HS-induced respiratory alkalosis. Chromium tended to increase average daily gain (0.86 vs. 0.95 kg, P = 0.070) during the 14-d supplementation under TN conditions before thermal exposure, which might be associated with the potential of Cr in improving overall insulin sensitivity, as evidenced by a reduced insulin resistance index calculated by Homeostatic Model Assessment (HOMA-IR; 0.65 vs. 0.51, P = 0.013) and a tendency of reduced fasting plasma insulin concentration (1.97 vs. 1.67 µU/mL, P = 0.094). Heat stress decreased the acute insulin releasing rate (P = 0.012) and consequently slowed glucose clearance rate (P = 0.035) during IVGTT. Besides, HS enlarged the values of area under the curve of NEFA during IVGTT (P < 0.01), indicating a reduced lipid mobilization. In conclusion, HS reduced insulin response to IVGTT. Chromium supplementation exhibited a potential in improving insulin sensitivity and mitigating HS symptoms in growing pigs.

High dietary selenium and vitamin E supplementation ameliorates the impacts of heat load on oxidative status and acid-base balance in sheep.

The objective of this study was to determine the efficacy of supranutritional dietary selenium and vitamin E (Vit E) to ameliorate the effect of heat stress (HS) on oxidative status and acid-base balance in sheep. Thirty-two Merino × Poll Dorset ewes were acclimated to indoor individual pen feeding of a pelleted control diet (0.24 g Se and 10 IU of Vit E/kg DM) for 1 wk. Sheep were then moved to metabolism cages in climatic chambers and randomly allocated to a 2 × 2 × 2 factorial design with the respective factors being dietary Se (0.24 and 1.20 mg/kg DM as Sel-Plex; Alltech, Australia), Vit E (10 and 100 IU/kg DM), and temperature for 2 wk. After 1 wk of acclimation in metabolic cages, 1 climatic chamber continued on thermoneutral (TN) conditions (18°C to 21°C and 40% to 50% relative humidity [RH]), and the other one was set to HS conditions (28°C to 40°C and 30% to 40% RH) for 1 wk. The sheep were then returned to individual pens and fed the control diet for 1 wk before being returned to the same diet as in the first period but a reversed thermal treatment for a further 2 wk. Physiological parameters were recorded 3 times daily, and blood samples were collected on d 1 and 7 of thermal treatment. Average respiration rate and rectal temperature of sheep were increased (P < 0.001) during HS; however, combined supranutritional supplementation of Se and Vit E reversed the effects of HS. Sheep given the high Se and high Vit E diet had a lower respiration rate (191 vs. 232 breaths/min; P = 0.012) and rectal temperature (40.33°C vs. 40.58°C; P = 0.039) under peak HS (1700 h) compared with those fed the low Se and low Vit E diet. Plasma reactive oxygen metabolites concentrations were reduced (P = 0.048) by 20%, whereas biological antioxidant potential was increased (P = 0.17) by 10% in sheep fed the high Se and high Vit E diet compared with those fed the low Se and low Vit E diet. Blood pH was elevated (P = 0.007) and bicarbonate was reduced (P = 0.049) under HS, and again, these effects were ameliorated by the high Se and high Vit E diet. Both white blood cell glutathione peroxidase gene expression and red blood cell lysate glutathione peroxidase activity were increased in sheep fed the high Se and high Vit E diet. These data suggest that supranutritional dietary Se or Vit E can reduce some of the negative effects of HS. However, the synergism between the 2 antioxidants improves their potential to ameliorate the impacts of HS in sheep.

Prediction of recurrence-free survival using a protein expression-based risk classifier for head and neck cancer.

Loco-regional recurrence in 50% of oral squamous cell carcinoma (OSCC) patients poses major challenge for oncologists. Lack of biomarkers that can predict disease aggressiveness and recurrence risk makes the scenario more dismal. On the basis of our earlier global proteomic analyses we identified five differentially expressed proteins in OSCC. This study aimed to develop protein biomarkers-based prognostic risk prediction model for OSCC. Sub-cellular expression of five proteins, S100A7, heterogeneous nuclear ribonucleoproteinK (hnRNPK), prothymosin α (PTMA), 14-3-3ζ and 14-3-3σ was analyzed by immunohistochemistry in test set (282 Indian OSCCs and 209 normal tissues), correlated with clinic-pathological parameters and clinical outcome over 12 years to develop a risk model for prediction of recurrence-free survival. This risk classifier was externally validated in 135 Canadian OSCC and 96 normal tissues. Biomarker signature score based on PTMA, S100A7 and hnRNPK was associated with recurrence free survival of OSCC patients (hazard ratio=1.11; 95% confidence interval 1.08, 1.13, P<0.001, optimism-corrected c-statistic=0.69) independent of clinical parameters. Biomarker signature score stratified OSCC patients into high- and low-risk groups with significant difference for disease recurrence. The high-risk group had median survival 14 months, and 3-year survival rate of 30%, whereas low-risk group survival probability did not reach 50%, and had 3-year survival rate of 71%. As a powerful predictor of 3-year recurrence-free survival in OSCC patients, the newly developed biomarkers panel risk classifier will facilitate patient counseling for personalized treatment.

Dietary antioxidants at supranutritional doses modulate skeletal muscle heat shock protein and inflammatory gene expression in sheep exposed to heat stress.

The objective of this study was to investigate the effects of chronic heat (thermal) stress and dietary antioxidant supplementation on the expression of heat shock proteins and inflammatory genes in the skeletal muscle of sheep. Twenty-four Merino × Poll Dorset crossbred ewes were allocated to either a control (10 IU vitamin E and 0.24 mg Se/kg DM) or high-antioxidant (VitE+Se; 100 IU vitamin E and 1.20 mg Se/kg DM) diet and were exposed to 2 thermal (temperature) treatments (thermoneutral [TN]: 18°C-21°C and 26%-30% relative humidity; heat stress [HS]: 28°C-40°C and 40%-50% relative humidity) for 1 wk. Physiological parameters were recorded daily, and muscle biopsies were conducted at the end of thermal treatments. Total RNA was extracted from muscle samples and reverse transcribed to cDNA for real-time PCR analysis. Respiration rates and rectal temperature were increased in response to HS (84.2 vs. 161 breaths per minute and 39.52°C vs. 40.06°C for TN and HS conditions, respectively; P < 0.001). There were interactions between dietary and thermal treatments, indicating that dietary antioxidant supplementation reduced respiration rate (P = 0.097) and rectal temperature (P = 0.086) of sheep during HS but not TN conditions. Skeletal muscle heat shock transcription factor 1 (HSF1) mRNA abundance was increased by HS (1.3-fold; P < 0.050) but was not changed (P = 0.77) by dietary antioxidant supplementation. The expression of skeletal muscle heat shock protein 70 (HSP70) mRNA was increased (P < 0.001) 3.5-fold by HS and tended (P = 0.08) to be increased by dietary antioxidant supplementation. Although there were no main effects of diet (P = 0.42) or HS (P = 0.47) on skeletal muscle HSP90 mRNA expression, there was an interaction (P = 0.040) such that HSP90 mRNA expression was increased (P = 0.010) in antioxidant-supplemented sheep under HS compared to TN conditions. Skeletal muscle nuclear factor kappa B (NF-κB) and tissue necrosis factor α (TNF-α) mRNA abundances were increased by exposure to heat (5.2-fold, P = 0.005 for NF-κB; 5.7-fold, P = 0.013 for TNF-α) ,but there was no main effect (P > 0.05) of dietary antioxidant supplementation. There was, however, an interaction between thermal and dietary treatments such that dietary antioxidant supplementation ameliorated the effect of HS on NF-κB and TNF-α mRNA expression. Taken together, these results indicate that high dietary antioxidants modulate skeletal muscle expression of heat shock proteins, proinflammatory cytokine, and NF-κB transcription, which may protect against HS in sheep.

Dietary antioxidants at supranutritional doses improve oxidative status and reduce the negative effects of heat stress in sheep.

The present study was undertaken to investigate the impact of heat (thermal) stress and dietary antioxidant supplementation on the oxidative and physiological status of sheep. Twenty-four Merino × Poll Dorset crossbred ewes were housed in 1 of 2 climatic chambers (thermoneutral or heat stress) and offered either a control (10 IU vitamin E/kg DM and 0.24 mg Se/kg DM) or high antioxidant (100 IU vitamin E/kg DM and 1.20 mg Se/kg DM) diet. The sheep were exposed to 2 thermal (temperature) treatments (thermoneutral [TN]: 18-21°C and 26-30% relative humidity; and heat stress [HS]: 28-40°C and 40-50% relative humidity) for 2 wk in a single reversal design. After 1 wk of dietary treatment, animals in 1 chamber were subjected to HS for 1 wk, with the temperature being increased to 40°C between 0900 and 1700 h and then maintained at 28°C overnight. Those sheep in the TN group were maintained at 18 to 21°C. Physiological parameters were recorded 4 times a day (0900, 1300, 1700, and 2100 h) and blood samples were collected on d 1 and 7 of heat treatment. Plasma samples and red blood cell lysates were assayed for oxidative stress biomarkers. The thermal treatments were then reversed and the above measures repeated. All measured physiological parameters were elevated (P < 0.001) by thermal treatment. Respiration rate was lower during HS in sheep supplemented with antioxidants as indicated by a diet × temperature × time interaction (P = 0.010). There was 13% decline (P = 0.014) in feed intake of the unsupplemented animals during HS whereas the same was maintained in sheep supplemented with high doses of antioxidants. Plasma reactive oxygen metabolites concentrations were reduced (114 vs. 85 units/dL; P < 0.005) while biological antioxidant potential tended to be increased (3,688 vs. 3,985 µmol/L; P = 0.070) in heat stressed sheep supplemented with antioxidants. The oxidative stress index was 30% lower (P < 0.001) in supplemented sheep (2.16 ± 0.06 arbitrary units) during HS than in unsupplemented sheep (3.12 ± 0.08 arbitrary units). Plasma advanced oxidation protein products tended (P = 0.070) to decrease in antioxidant supplemented heat stressed sheep as compared to their unsupplemented counterparts. It was concluded that heat stress negatively affects the oxidative status of sheep along with the physiological responses and some of these affects can be ameliorated through dietary antioxidants supplementation at supranutritional concentrations.

Regulation of heat shock proteins 27 and 70, p-Akt/p-eNOS and MAPKs by Naringin Dampens myocardial injury and dysfunction in vivo after ischemia/reperfusion.

Naringin has antioxidant properties that could improve redox-sensitive myocardial ischemia reperfusion (IR) injury. This study was designed to investigate whether naringin restores the myocardial damage and dysfunction in vivo after IR and the mechanisms underlying its cardioprotective effects. Naringin (20-80 mg/kg/day, p.o.) or saline were administered to rats for 14 days and the myocardial IR injury was induced on 15(th) day by occluding the left anterior descending coronary artery for 45 min and subsequent reperfusion for 60 min. Post-IR rats exhibited pronounced cardiac dysfunction as evidenced by significantly decreased mean arterial pressure, heart rate, +LVdP/dt max (inotropic state), -LVdP/dt max (lusitropic state) and increased left ventricular end diastolic pressure as compared to sham group, which was improved by naringin. Further, on histopathological and ultrastructural assessments myocardium and myocytes appeared more normal in structure and the infarct size was reduced significantly in naringin 40 and 80 mg/kg/day group. This amelioration of post-IR-associated cardiac injury by naringin was accompanied by increased nitric oxide (NO) bioavailability, decreased NO inactivation to nitrotyrosine, amplified protein expressions of Hsp27, Hsp70, ß-catenin and increased p-eNOS/eNOS, p-Akt/Akt, and p-ERK/ERK ratio. In addition, IR-induced TNF-α/IKK-ß/NF-κB upregulation and JNK phosphorylation were significantly attenuated by naringin. Moreover, western blotting and immunohistochemistry analysis of apoptotic signaling pathway further established naringin cardioprotective potential as it upregulated Bcl-2 expression and downregulated Bax and Caspase-3 expression with reduced TUNEL positivity. Naringin also normalized the cardiac injury markers (lactate dehydrogenase and creatine kinase-MB), endogenous antioxidant activities (superoxide dismutase, reduced glutathione and glutathione peroxidase) and lipid peroxidation levels. Thus, naringin restored IR injury by preserving myocardial structural integrity and regulating Hsp27, Hsp70, p-eNOS/p-Akt/p-ERK signaling and inflammatory response.

Phosphorylation of Akt/GSK-3ß/eNOS amplifies 5-HT2B receptor blockade mediated anti-hypertrophic effect in rats.

Herein, we studied the cross talk between 5-HT(2B) receptor blocker (SB-204741) and GSK-3ß inhibitor (SB-216763) in isoproterenol-induced cardiac hypertrophy for 28 days. SB-204741 treatment significantly ameliorated (P<0.05) myocardial dysfunction, myocyte area, fibrosis and myocardial architecture in isoproterenol insulted myocardium. Moreover, this improvement in functional and morphological changes was associated with suppression of hypertrophic (BNP and CK-MB), inflammatory (IKK-ß/NF-κB/TNF-α and CRP), and apoptotic markers (TUNEL positivity and Bax expression) along with phosphorylation of Akt/GSK-3ß/ß-catenin/eNOS. Intriguingly, co-treatment with GSK-3ß inhibitor (P<0.01) further amplified the anti-hypertrophic effect of SB-204741 (P<0.05) such that the effect was indistinguishable from that of vehicle treated rats. Thus, 5-HT(2B) receptor blockade mediated anti-hypertrophic effect is atleast in part is governed through phosphorylation of Akt/GSK-3ß/ß-catenin/eNOS via attenuating inflammatory and apoptotic pathways.

Guggulsterone (GS) inhibits smokeless tobacco and nicotine-induced NF-κB and STAT3 pathways in head and neck cancer cells.

Understanding the molecular pathways perturbed in smokeless tobacco- (ST) associated head and neck squamous cell carcinoma (HNSCC) is critical for identifying novel complementary agents for effective disease management. Activation of nuclear factor-kappaB (NF-κB) and cyclooxygenase-2 (COX-2) was reported in ST-associated HNSCC by us [Sawhney,M. et al. (2007) Expression of NF-kappaB parallels COX-2 expression in oral precancer and cancer: association with smokeless tobacco. Int. J. Cancer, 120, 2545-2556]. In search of novel agents for treatment of HNSCC, we investigated the potential of guggulsterone (GS), (4,17(20)-pregnadiene-3,16-dione), a biosafe nutraceutical, in inhibiting ST- and nicotine-induced activation of NF-κB and signal transducer and activator of transcription (STAT) 3 pathways in HNSCC cells. GS inhibited the activation of NF-κB and STAT3 proteins in head and neck cancer cells. This inhibition of NF-κB by GS resulted from decreased phosphorylation and degradation of nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha the inhibitory subunit of NF-κB. Importantly, treatment of HNSCC cells with GS abrogated both ST- and nicotine-induced nuclear activation of NF-κB and pSTAT3 proteins and their downstream targets COX-2 and vascular endothelial growth factor. Furthermore, GS treatment decreased the levels of ST- and nicotine-induced secreted interleukin-6 in culture media of HNSCC cells. In conclusion, our findings demonstrated that GS treatment abrogates the effects of ST and nicotine on activation of NF-κB and STAT3 pathways in HNSCC cells that contribute to inflammatory and angiogenic responses as well as its progression and metastasis. These findings provide a biologic rationale for further clinical investigation of GS as an effective complementary agent for inhibiting ST-induced head and neck cancer.

Prognostic significance of nuclear pSTAT3 in oral cancer.

BACKGROUND: Aberrant nuclear accumulation of proteins influences tumor development and may predict biologic aggressiveness and disease prognosis. This study determined the prognostic significance of pSTAT3 (phosphorylayed signal transducer and activator of transcription 3) in oral squamous cell carcinomas (OSCCs). METHODS AND RESULTS: Using immunohistochemistry, a significant increase in nuclear accumulation of pSTAT3 was observed in 49 of 90 leukoplakias (54.4%) and 63/94 OSCCs (67%) (p(trend) < .001). Increased pSTAT3 was associated with tumor stage (p = .01), nodal metastasis (p = .0018), and tobacco consumption (p = .004). Kaplan-Meier analysis demonstrated that OSCC with increased nuclear pSTAT3 showed significantly reduced disease-free survival (13 months), compared with the patients with no nuclear pSTAT3 expression (64 months, p = .019). Cox regression analysis revealed nuclear pSTAT3 as the most significant predictor of poor prognosis (p = .024, hazard ratio [HR] = 2.7). CONCLUSIONS: Increased nuclear accumulation of pSTAT3 occurs in early premalignant stages and is a marker for poor prognosis of OSCC.

Studies on production performance and toxin residues in tissues and eggs of layer chickens fed on diets with various concentrations of aflatoxin AFB1.

1. An experiment on 1-week-old, White Leghorn female chicks was conducted to study the effect of aflatoxin AFB1 on weight gain, feed intake, feed gain ratio, age at sexual maturity, production and quality of eggs, retention of nutrients, pathoanatomical and histopathological parameters, and also on AFB1 residues in eggs and muscles of hens. The chicks were assigned to 4 dietary treatments: D1 (without AFB1), D2 (2.50 mg/kg AFB1), D3 (3.13 mg/kg AFB1), D4 (3.91 mg/kg AFB1) up to the age of 40 weeks. 2. At the end of the experiment, the mean body weight gain and feed intake were significantly lower in all aflatoxin-fed groups compared to control. The feed gain ratios were noted as 13.41, 13.59, 13.82 and 14.71, with the group fed the highest concentration of AFB1 showing a significantly poorer ratio than other groups. 3. Age at sexual maturity was also affected adversely by dietary AFB1: 193 d for D4 as compared to as early as 148 d for D1. Hen-d egg production was recorded as 96.92, 74.67, 65.98 and 50.75 in D1, D2, D3 and D4, respectively. 4. Average egg weights at the end of the experiment were 57.77, 57.49, 57.54 and 54.66 for D1, D2, D3 and D4, respectively. Shape index was significantly lower in D4 as compared to control. Contrary to this, albumen index was significantly higher in D4 as compared to D1. The values of yolk indices and eggshell thickness did not differ significantly among treatment groups. However, colour of yolk was reduced in all aflatoxin-fed groups compared to control. 5. Retentions of dry matter, crude protein, ether extract, calcium and metabolisable energy were adversely affected at various levels of AFB1 compared to control. 6. Pathoanatomical and histopathological studies showed various adverse changes in liver, kidney, heart, ovaries and bursa of Fabricius in AFB1-fed groups. 7. Different amounts of aflatoxin residues were detected in eggs and breast muscles of hen in all AFB1-fed groups.

Transcriptional upregulation of human cathepsin L by VEGF in glioblastoma cells.

The role of vascular endothelial growth factor (VEGF) on cathepsin L expression was investigated in human glioblastoma cells (U87MG). Our results demonstrate the transcriptional upregulation of cathepsin L expression by VEGF. Transient transfection of U87MG cells with VEGF expression vector significantly increased cathepsin L activity. These results were further corroborated by a parallel increase in the mRNA levels and promoter activity of cathepsin L by VEGF. By deletion analysis, we identified a 47 base pair VEGF response element (VRE) in human cathepsin L promoter. Site directed mutagenesis studies demonstrated that both SP-1 and AP-4 motifs present in this region contribute to VEGF responsiveness. These results prove for the first time that over-expression of VEGF in human glioblastoma cells induces cathepsin L expression at the transcriptional level. This mechanism could be involved in the enhanced tumorogenic potential of these cells.

Tropical pyomyositis (myositis tropicans): current perspective.

Tropical pyomyositis, a disease often seen in tropical countries, is characterised by suppuration within skeletal muscles, manifesting as single or multiple abscesses. The most common organism implicated is Staphylococcus aureus. In 20%-50% of cases there is a history of trauma to the affected muscles. Commonly involved muscles are quadriceps, glutei, pectoralis major, serratus anterior, biceps, iliopsoas, gastrocnemius, abdominal and spinal muscles. Early diagnosis is often missed because of lack of specific signs, unfamiliarity with the disease, atypical manifestations, and a wide range of differential diagnosis. Diagnostic techniques like ultrasound and computed tomography/magnetic resonance imaging are very useful in diagnosis. The diagnosis is confirmed either by biopsy or aspiration of pus from the affected muscles. The initial antibiotic of choice is cloxacillin. Incision and drainage are important components of management. Treatment for Gram negative or anaerobic organisms should be instituted, whenever indicated. Physicians should become more familiar with this potentially life threatening but curable infective disease entity.