Chemometric Based Identification and Validation of Specific Chemical Markers for Geographical, Seasonal and Gender Variations in Tinospora cordifolia Stem using HPLC-ESI-QTOF-MS Analysis.

INTRODUCTION: Tinospora cordifolia is a widely distributed medicinal plant used in various traditional and commercial Ayurvedic formulations. Due to the wide use of this plant it is important to know the extent of variability in the metabolite profile resulting from geographical location, season and gender. OBJECTIVE: To develop a statistical approach based on phytochemical markers for confident prediction of variations in metabolic profile and cytotoxicity due to geographical, seasonal and gender difference in T. cordifolia stem. METHODS: A HPLC-ESI-QTOF-MS method was used for the metabolite profiling of T. cordifolia stem. The data were analysed using chemometric methods including Student's t-test, ANOVA, FA/PCA and ROC curve analysis and validated for the identification of chemical variations. The bioactivity of selected samples was also tested using a cell cytotoxicity assay to assess the functional aspect of the phytochemical variability. RESULTS: The chemometric approach applied here identified marker ions for geographical locations (m/z 294.1139 and 445.2136), seasons (m/z 344.1482, 359.1501, and 373.1305) and gender (m/z 257.1380) with 100% statistical sensitivity and specificity. An in vitro cytotoxicity evaluation revealed that male T. cordifolia stem was the most effective in inhibiting the growth of cancerous cell lines. CONCLUSIONS: The developed and validated chemometric approach identified the analytical markers for phytochemical variations in unknown T. cordifolia stem samples from male or female plants and samples collected from different geographical locations and seasons. The results are supported by comparative cytotoxic activity data. Copyright © 2017 John Wiley & Sons, Ltd.

Analysis of phytochemical variations in dioecious Tinospora cordifolia stems using HPLC/QTOF MS/MS and UPLC/QqQLIT -MS/MS.

INTRODUCTION: The stem of dioecious Tinospora cordifolia (Menispermaceae) is a commonly used traditional Ayurvedic medicine in India having several therapeutic properties. OBJECTIVE: To develop and validate LC-MS methods for the identification and simultaneous quantitation of various secondary metabolites and to study metabolomic variations in the stem of male and female plants. METHODS: Ethanolic extract of stems were analysed by HPLC/ESI-QTOF-MS/MS for rapid screening of bioactive phytochemicals. High resolution MS and MS/MS in positive ESI mode were used for structural investigation of secondary metabolites. An UPLC/ESI-QqQ(LIT) -MS/MS method in MRM mode was developed and validated for the simultaneous quantitation of five bioactive alkaloids. RESULTS: Identification and characterisation of 36 metabolites including alkaloids, sesquiterpenes and phytoecdysteroids were performed using LC-MS and MS/MS techniques. The bioactive alkaloids such as jatrorrhizine, magnoflorine, isocorydine, palmatine and tetrahydropalmatine were successfully quantified in male and female plants. The mean abundances of magnoflorine jatrorrhizine, and oblongine were significantly (P < 0.05) higher in male plants while mean abundances of tetrahydropalmatine, norcoclaurine, and reticuline were significantly (P < 0.05) higher in female plants. CONCLUSIONS: Phytochemicals in the stem of male and female Tinospora cordifolia showed significant qualitative and quantitative variations. LC-MS and MS/MS methods can be used to differentiate between male and female plants based on their chemical profiles and quantities of the marker bioactive alkaloids. This chemical composition difference was also evident during vegetative stage when there were no male and female flowers.

TNF-α gene expression in subcutaneous adipose tissue associated with HOMA in Asian Indian postmenopausal women.

The present study determines the fat depot-specific expression of leptin and TNF-α and its association with biochemical parameters in postmenopausal women. A total of 108 postmenopausal women were recruited prospectively; 54 were with metabolic syndrome (cases) and 54 were without metabolic syndrome (controls). Leptin and TNF-α mRNA expression in visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) were done by Real Time-RT PCR. In cases, the mean (±SD) serum estrogen was significantly lower (41.33±24.90 vs. 23.95±14.45, p<0.001) while leptin (12.85±4.51 vs. 10.34±3.89, p=0.002) and TNF-α (13.81±7.13 vs. 8.00±4.38, p<0.001) were significantly higher as compared to controls. Further, the mean relative VAT mRNA expression of both leptin (0.33±0.29 vs. 0.05±0.09, p<0.001) and TNF-α (0.32±0.31 vs. 0.13±0.09, p<0.001) and expression of SAT leptin (4.91±4.01 vs. 0.50±0.92, p<0.001) also lowered significantly in cases as compared to controls. Further, the relative VAT expression of both leptin (r=-0.32, p<0.001) and TNF-α (r=-0.23, p<0.01) showed significant and negative correlation with glucose; expression of SAT leptin showed significant and positive correlation with HDL (r=0.20, p<0.05) and serum estrogen (r=0.30, p<0.01) while negative correlation with glucose (r=-0.26, p<0.01) and serum TNF-α (r=-0.29, p<0.01); and expression of SAT TNF-α showed significant and positive correlation with insulin (r=0.21, p<0.05) and HOMA (r=0.20, p<0.05). In conclusion, the VAT and SAT leptin mRNA expressions may have a modulatory role in metabolic syndrome.

CD3+, CD4+ & CD8+ tumour infiltrating lymphocytes (TILs) are predictors of favourable survival outcome in infiltrating ductal carcinoma of breast.

BACKGROUND & OBJECTIVES: Tumour infiltrating lymphocytes (TILs) represent the host immune response against cancer cells associated with good or bad prognosis in different tumour types. This study was undertaken to evaluate the significance of CD3+, CD4+ and CD8+ TILs in breast cancer tissues in relation to clinico-pathological variables and survival outcome. METHODS: Immunohistochemistry (IHC) was performed with antibodies against CD3, CD4 and CD8 antigens on formalin-fixed paraffin-embedded tissue sections of 150 breast cancer patients. Intratumoural and stromal TIL counting was performed semiquantitatively. RESULTS: The higher CD3+, CD4+ and CD8+ intratumoural and stromal counts showed independent and direct association with good prognosis. The prognostic predictor value of intratumoural counts was higher than stromal counts. The independent associations of intratumoural and stromal counts became more prominent when adjusted with stage and grade, respectively. Among intratumoural counts, the high (++/+++) CD4+ count (OR=3.85, 95% CI=3.28-16.71, P<0.001) showed the highest survival followed by CD3+ (OR=2.70, 95% CI=1.76-8.30, P=0.001) and CD8+ (OR=2.58, 95% CI=1.55-5.86, p0 =0.001) the least when compared to respective low (+) counts. In contrast, among stromal counts, the high CD8+ count (OR=3.13, 95% CI=2.20-9.57, p0 <0.001) showed the highest survival followed by CD4+ (OR=3.02, 95% CI=2.07-8.89, p0 <0.001) and CD3+ (OR=2.45, 95% CI=1.53-6.73, p0 =0.002) the least. INTERPRETATION & CONCLUSIONS: Our results suggest that intratumoural CD4+ and stromal CD8+ counts by immunohistochemistry may serve as an independent prognosticator for favourable outcome in breast cancer.

Tracing the tracks of genotoxicity by trivalent and hexavalent chromium in Drosophila melanogaster.

Mutagen sensitive strains (mus) in Drosophila are known for their hypersensitivity to mutagens and environmental carcinogens. Accordingly, these mutants were grouped in pre- and post-replication repair pathways. However, studying mutants belonging to one particular repair pathway may not be adequate for examining chemical-induced genotoxicity when other repair pathways may neutralize its effect. To test whether both pre-and post-replication pathways are involved and effect of Cr(III)- and Cr(VI)-induced genotoxicity in absence or presence of others, we used double mutant approach in D. melanogaster. We observed DNA damage as evident by changes in Comet assay DNA migration in cells of larvae of Oregon R(+) and single mutants of pre- (mei-9, mus201 and mus210) and post- (mei-41, mus209 and mus309) replication repair pathways and also in double mutants of different combinations (pre-pre, pre-post and post-post replication repair) exposed to increasing concentrations of Cr(VI) (0.0, 5.0, 10.0 and 20.0 µg/ml) for 48 h. The damage was greater in pre-replication repair mutants after exposure to 5.0 µg/ml Cr(VI), while effects on Oregon R(+) and post replication repair mutants were insignificant. Post-replication repair mutants revealed significant DNA damage after exposure to 20.0 µg/ml Cr(VI). Further, double mutants generated in the above repair categories were examined for DNA damage following Cr(VI) exposure and a comparison of damage was studied between single and double mutants. Combinations of double mutants generated in the pre-pre replication repair pathways showed an indifferent interaction between the two mutants after Cr(VI) exposure while a synergistic interaction was evident in exposed post-post replication repair double mutants. Cr(III) (20.0 µg/ml) exposure to these strains did not induce any significant DNA damage in their cells. The study suggests that both pre- and post-replication pathways are affected in Drosophila by Cr(VI) leading to genotoxicity, which may have consequences for metal-induced carcinogenesis.

Assessment of urban air pollution and it's probable health impact.

The present study deals with the quantitative effect of vehicular emission on ambient air quality during May 2006 in urban area of Lucknow city. In this study SPM, RSPM, SO2, NOx and 7 trace metals associated with RSPM were estimated at 10 representative locations in urban area and one village area for control. Beside this, air quality index (AQI), health effects of different metals and mortality were assessed. The 24 hr average concentration of SPM, RSPM, SO2 and NOx was found to be 382.3, 171.5, 24.3 and 33.8 microg m(-3) respectively in urban area and these concentrations were found to be significantly (p < 0.01) higher by 94.8, 134.8, 107.4 and 129.6% than control site respectively The 24 hr mean of SPM and RSPM at each location of urban area were found to be higher than prescribed limit of National Ambient Air Quality Standard (NAAQS) except SPM for industrial area. The 24 hr mean concentration of metals associated with RSPM was found to be higher than the control site by 52.3, 271.8, 408.9, 75.81, 62.7, 487.54 and 189.5% for Fe, Cu, Pb, Zn, Ni, Mn and Cr respectively. The inter correlation of metals Pb with Mn, Fe and Cr; Zn with Ni and Cr; Ni with Cr; Mn with Fe and Cu with Cr showed significant positive relation either at p < 0.05 or p < 0.01 level. Metals Pb, Mn and Cr (p < 0.01) and Cu (p < 0.05) showed significant positive correlation with RSPM. These results indicate that ambient air quality in the urban area is affected adversely due to emission and accumulation of SPM, RSPM, SO2, NOx and trace metals. These pollutants may pose detrimental effect on human health, as exposure of these are associated with cardiovascular and respiratory diseases, neurological impairments, increased risk of preterm birth and even mortality and morbidity.

Fine particles (PM2.5) in ambient air of Lucknow city due to fireworks on Diwali festival.

People burn crackers world over on different occasions in different countries to express their happiness. Fireworks in large amounts aggravate the level of air pollutants and cause significant short-term air quality degradation with possible impact on human health. Fine particles (PM2.5 < or = 2.5 microm), which may pose detrimental effects on human health and ecosystems were monitored in a residential area of Lucknow city to assess the elevated level due to bursting of firecrackers during Diwali festival. The 24 hr mean PM2.5 of normal day, pre Diwali day, Diwali day and post Diwali day was found to be 124, 154, 352 and 174 microg m(-3) respectively and much above the US-EPA limit (65 microg m(-3)). The 12 hr mean concentration of PM2.5 on Diwali night (591 microg m(-3)) increased 3.9 fold than the respective night of normal day (159 microg m(-3)) and was significantly higher (p<0.01) than normal day and pre and post Diwali night. Mean comparison showed that Diwali day was significantly (p<0.01) different from others (except post Diwali day) and for this high accumulation during night time, after fireworks (suspension) was found to be more responsible than the period of lighting of crackers (formation). This study indicated that there is high accumulation of PM2.5 generated due to fireworks on Diwali festival which remains suspended in the air for up to 20 hr During this period, extra mass burden of 289 microg m(-3) equivalent to 1.9 normal day (of this study) was imposed in the environment. The short-term high accumulation of PM2.5 is a matter of serious concern for city dwellers as it can penetrate deep into the lungs and cause many respiratory and cardiovascular diseases.

Metals concentration associated with respirable particulate matter (PM10) in industrial area of Eastern U.P. India.

The present study deals with the assessment of ambient air quality with respect to respirable suspended particulate matter (RSPM or PM10 < or = 10 microm) and trace metals (Fe, Zn, Cu, Cr Ni, Cd, Mn and Pb) concentrations in RSPM at five locations of Renukoot, an industrial area of Eastern Uttar Pradesh. The 24 hr mean concentrations of PM10 ranged between 69.3 to 118.9 microg m(-3), which is well within the permissible limit (150 microg m(-3)) of national ambient air quality standards (NAAQS) but found higher than the prescribed annual daily limit of US EPA (50 microg m(-3)). The ambient air was mostly dominated by the Fe and least by the Cd among the metal analysed. Murdhawa, a commercial place influenced by vehicular population, is found to be the most polluted area of Renukoot and Dongia nalla (forest area) the least. The ambient air of Murdhawa is rich in Cu and Ni, indicating contribution of mobile sources. The Rammandir a residential place near the industry, is rich in Cd and Cr suggesting contribution of point sources. The Ni concentration is found to be alarmingly high in the air at all the locations except Dongia nallah, when compared with the EC (European Commission) limit (20 ng m(-3)). The Cd concentration is found to be higher only at Rammandir as compared with the EC limit (5 ng m(-3)). Mean concentrations of Zn, Pb and Mn are found to be almost equal in the ambient air of all the locations, suggesting the significance of sources contributing to presence of these metals. Zn, Cu, Pb and Ni having a significant correlation with PM10 indicate the same source contributing these metals as well as PM10. The present study has focused on the quantitative variation in different metals in the PM10, which is extremely harmful due to their toxic and carcinogenic nature.

Anthropometric study using craniofacial features to determine gender in Lucknow population.

BACKGROUND: Gender is one of the main characteristics analyzed for positive human identification in forensic medicine. The methods involving physical anthropology present high rate of accuracy for human identification and gender estimation. AIM: This study aimed to determine gender through different craniofacial variables using physical anthropometric methods. MATERIALS AND METHODS: A cross-sectional study was conducted among 100 individuals (50 males and 50 females) in Lucknow. Variables studied through physical anthropometry in both the genders were facial height, nasion-to-menton distance, interzygomatic arch width, and intercanthal width using a digital sliding caliper. All the measurements were taken twice. The final value was the average of the two obtained values. RESULTS: Comparing the mean craniofacial features between two genders, t-test revealed significantly higher facial height, pronasale-to-menton distance, and interzygomatic width in males as compared to females, but the mean intercanthal width was found to be the same. Pearson's correlation analysis revealed a positive correlation between facial height and pronasale-to-menton distance, facial height and interzygomatic width, pronasale-to-menton distance and interzygomatic width, and interzygomatic width and intercanthal width. CONCLUSION: The craniofacial features may serve as diagnostic markers for gender identification and can be used interchangeably.

Physical interaction of estrogen receptor with MnSOD: implication in mitochondrial O2.- upregulation and mTORC2 potentiation in estrogen-responsive breast cancer cells.

Augmented reactive oxygen species levels consequential to functional alteration of key mitochondrial attributes contribute to carcinogenesis, either directly via oxidative DNA damage infliction or indirectly via activation of oncogenic signaling cascades. We previously reported activation of a key oncogenic signaling cascade via mammalian target of rapamycin (mTOR) signaling complex-2 (mTORC2) owing to estrogen receptor (ER-α)-dependent augmentation of O2.- within the mitochondria of 17-ß-estradiol (E2)-stimulated breast cancer cells. Manganese superoxide dismutase (MnSOD) is the principal mitochondrial attribute governing mitochondrial O2.- homeostasis, raising the possibility that its functional alteration could be instrumental in augmenting mitochondrial O2.- levels in breast cancer cells. Here we show ER-dependent transient inhibition of MnSOD catalytic function in breast cancer cells. Catalytic function of MnSOD is tightly regulated at the post-translational level. Post-translational modifications such as phosphorylation, nitration and acetylation represent key regulatory means governing the catalytic function of MnSOD. Acetylation at lysine-68 (K68) inhibits MnSOD catalytic activity and thus represents an important post-translational regulatory mechanism in human cells. Using reciprocal immunoprecipitation and proximity ligation assay, we demonstrate the occurrence of direct physical interaction between ER-α and MnSOD in human breast cancer cells, which in turn was associated with potentiated acetylation of MnSOD at K68. In addition, we also observed diminished interaction of MnSOD with sirtuin-3, the key mitochondrial deacetylase that deacetylates MnSOD at critical K68 and thereby activates it for scavenging O2.-. Consequently, compromised deacetylation of MnSOD at K68 leading to its inhibition and a resultant buildup of O2.- within the mitochondria culminated in the activation of mTORC2. In agreement with this, human breast cancer tissue specimen exhibited a positive correlation between acetyl-MnSODK68 levels and phospho-Ser2481 mTOR levels. In addition to exposing the crosstalk of ER-α with MnSOD post-translational regulatory mechanisms, these data also unravel a regulatory role of ER/MnSOD interaction as an important control switch for redox regulation of ER-α-responsive oncogenic signaling cascades. Furthermore, our study provides a mechanistic link for ER-α-dependent O2.- potentiation and resultant mTORC2 activation in breast cancer cells.

Gefitinib, Methotrexate and Methotrexate plus 5-Fluorouracil as palliative treatment in recurrent head and neck squamous cell carcinoma.

This study compared the efficacy and toxicity of Gefitinib, Methotrexate and Methotrexate plus 5-Fluorouracil (5-FU) in patients of recurrent squamous cell carcinoma of head and neck (SCCHN) treated with palliative intent. Patients with recurrent SCCHN not amenable to curative treatment were randomly assigned to Gefitinib, Methotrexate or Methotrexate plus 5-FU arm. The primary end point was overall survival. Secondary end points of interest were objective response rate, toxicity and quality of life. Total 117 patients were analyzed. Median overall survival and objective response rates were 8.8 months, 7.8 months and 8.1 months and 7.7%, 5.0% and 7.9% in Gefitinib, Methotrexate and Methotrexate plus 5-FU arms respectively with no statistically significant difference between 3 arms. Gefitinib had different toxicity profile compared with other arms. Majority of toxicities were Grade 1 or Grade 2. Gefitinib had significant improvement in quality of life during initial months over Methotrexate. There was no suggestion that Gefitinib significantly prolonged overall survival compared with Methotrexate and Methotrexate plus 5-FU. However, improved Quality of Life with manageable toxicities was observed.

Prognostics of Cyclin-D1 expression with chemoradiation response in patients of locally advanced oral squamous cell carcinoma.

OBJECTIVE: Cyclin-D1 has been strongly implicated in cell cycle proliferation particularly in the G1/S checkpoint in the cell cycle, and prognosis in many human malignancies. The present study evaluates its prognostic significance with chemoradiation response in patients of locally advanced oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: A total of 97 OSCC patients (females = 19 and males = 78), aged 20-67 years and stage III/IV were recruited. Treatment response was assessed according to World Health Organization criteria. Cyclin-D1 expression in tumor tissue was estimated by immunohistochemical method and quantified as percentage positive nuclei. RESULTS: The Cyclin-D1 expression showed significant (P < 0.01 or P < 0.001) association with tumor size, lymph node status, and clinical stage. After chemoradiation, there were 53.6% complete response (CR) and 34.0% partial response (PR) in primary tumor, and 49.5% CR and 39.2% PR in lymph node; giving an overall response rate of 85.6%. Further, the mean Cyclin-D1 expression showed significant (P < 0.05 or P < 0.001) and inverse association with chemoradiation responses (tumor size, lymph node status and overall treatment response). The 2-year progression-free and overall survival (OS) was 95.89% and 83.31% respectively. Multivariate Cox regression analysis found site of primary tumor, clinical stage, and Cyclin-D1 expression the significant (P < 0.05 or P < 0.01) and independent prognostic markers of OS and among these Cyclin-D1 expression showed the worst prognosis. The high Cyclin-D1 expression (>50%) also showed significantly lower survival in OSCC patients when compared with those had low (<10%) and moderate expressions (10-50%) (Logrank test: χ(2) = 44.42, P < 0.001). CONCLUSION: The high Cyclin-D1 expression may serve as a poor prognostic marker in OSCC.

Resistin 420C/G gene polymorphism on circulating resistin, metabolic risk factors and insulin resistance in adult women.

OBJECTIVE: To investigate the frequency association between resistin gene polymorphism with its circulating levels, metabolic risk factor and insulin resistance in adult women. DESIGN: Totally 615 subjects were enrolled for the study, 305 women were with metabolic syndrome and 310 women were without metabolic syndrome according to NCEP-ATP III criteria. Fasting circulatory level of resistin, insulin, plasma glucose and lipid profiles were estimated along with calculation of insulin resistance. Resistin 420C/G promoter region polymorphism was done by RFLP method. RESULTS: Variant genotype (CC vs CG+GG) (p<0.001: OR=2.22: 95% CI=1.60-3.10) of 420C/G resistin gene polymorphism was less frequently observed in control population. Further dividing subjects into two groups according to absence (Resistin -1) or presence (Resistin-2) of the G allele, significantly high levels of triglyceride (p<0.001), plasma glucose (p=0.012), systolic blood pressure (p<0.001), diastolic blood pressure (p<0.001), waist hip ratio (p<0.001), body mass index (p<0.001) and resistin (p<0.001), were observed in resistin-2 group. CONCLUSION: Present study shows that 420C/G polymorphism of resistin gene directly correlated to its high circulating level and metabolic risk factors, specifically markers of obesity and atherosclerosis, so it may have an important role in the development of metabolic syndrome and cardio metabolic diseases.

Clinical benefits of concurrent capecitabine and cisplatin versus concurrent cisplatin and 5-flurouracil in locally advanced squamous cell head and neck cancer.

We aimed to assess the efficacy and safety of concurrent capecitabine and cisplatin over concurrent cisplatin and 5-flurouracil (5-FU) in locally advanced squamous cell carcinoma of the head and neck. One hundred and fifty-three patients (all of whom had stage III or IV unresectable disease with no distant metastases and who had received two cycles of taxol and cisplatin chemotherapy) were randomly assigned to receive either concurrent cisplatin (75 mg/m(2) in day 1 and 2) and 5-FU (750 mg/m(2) in day 1, 2, and 3) from the first day of radiotherapy at an interval of 3 weeks (Arm I) or cisplatin (75 mg/m(2) in day 1 and 2) and capecitabine (750 mg/m(2) in two divided doses from day 1-14) from the first day of radiotherapy at a 3-week interval (Arm II). Results showed that patients in Arm II had a significantly better rate of complete response, fewer nodes, and better overall response compared to those in Arm I. The two groups had a similar 3-year disease-free survival, progression free survival, and overall survival, i.e. they did not differ significantly. Variables indicating the quality of life of the two groups were compared. Patients in Arm II had a significantly higher quality of life compared to those in Arm I. The two groups had similar treatment-related acute and late toxicity, i.e. they did not differ significantly. These results have thoroughly substantiated the contention that concurrent chemoradiation with capecitabine and cisplatin may be regarded as an effective and well-tolerated regimen in the treatment of the patients with locally advanced head and neck cancer.

2,3-Diaryl-2H-1-benzopyran derivatives interfere with classical and non-classical estrogen receptor signaling pathways, inhibit Akt activation and induce apoptosis in human endometrial cancer cells.

OBJECTIVES: The present study was undertaken to explore the mechanism of anti-proliferative action of benzopyran compound D1 (2-[piperidinoethoxyphenyl]-3-phenyl-2H-benzopyran) and its hydroxy-(D2) and methoxy-(D3) derivatives in Ishikawa and human primary endometrial adenocarcinoma cells. METHODS: Transcriptional activation assays were performed using luciferase reporter system and cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The stage of cell cycle was determined by flow-cytometry and real time analysis of cyclinE1 and cdc2 genes. The apoptotic effects were measured by AnnexinV/PI staining and TUNEL. The expression of PCNA, cyclinD1, pAkt, XIAP, cleaved caspase-9, -3, PARP, Bax and Bcl2 were determined by immunoblotting. The caspase-3 activity and mitochondrial membrane potential were measured by colorimetric assay. RESULTS: All three compounds inhibited E(2)-induced ERE- and AP-1-mediated transactivation and proliferation in endometrial adenocarcinoma cells dose-dependently. Compound D1 caused the arrest of cells in the G(2) phase while D2 and D3 caused arrest in G(1) phase of the cell cycle. All compounds interfered with Akt activation, decreased XIAP expression leading to an increased cleavage of caspase-9, -3, PARP, increased Bax/Bcl2 ratio and caspase-3 activity. CONCLUSION: Findings suggest that benzopyran derivatives inhibit cellular proliferation via modulating ER-dependent classical and non-classical signaling mechanisms, interfere with Akt activation and induce apoptosis via intrinsic pathway in endometrial adenocarcinoma cells.

Codon optimization of the major antigen encoding genes of diverse strains of influenza a virus.

A large number of influenza A virus outbreaks and mortality occurred in the world recently, an urgent attention to develop effective and sufficient quantity of vaccines are needed. Vaccines are generally protein with immunogenic properties and are not expressed in sufficient quantity because of the codon bias, so it is necessary to optimize its codon in the expression host. Codon optimization was used to improve the protein expression in living organisms by increasing the translational efficiency of gene of interest. Two surface antigenic glycoproteins, hemagglutinin (HA) and neuraminidase (NA) are present in influenza A viruses. We have used HA and NA genes from 19 strains of influenza A viruses for codon optimization in E. coli. Both genes of the influenza virus show that the codon adaptation index (CAI) and GC content of the genes in optimized DNA were enhanced significantly (p <0.01) as compared to wild type. CAI and GC of HA in optimized DNA was enhanced by 3.2 (68.5%) and 1.2 (16.2%) fold respectively, while in NA it was increased by 3.3 (69.7%) and 1.2 (15.8%) fold respectively. Our finding demonstrates that the optimized genes could be useful for better expression in host without any truncated proteins and also helpful for protein folding and function. This work provides new insight in the synthetic biology research.

Interleukin-6 G-174C gene polymorphism and serum resistin levels in North Indian women: potential risk of metabolic syndrome.

The present investigations were aimed to identify the possible association between genetic polymorphism in interleukin-6 (IL-6) G-174C gene, which confers susceptibility to metabolic syndrome, and serum level of resistin in North Indian women. The study population comprised 370 unrelated Indian women (192 having abdominal obesity and 178 controls). Polymorphism in genotype (CC+GC) of IL-6 G-174C gene was determined using a combination of polymerase chain reaction (PCR) and sequence-specific primer with restriction fragment length polymorphism (RFLP) technology. Insulin resistance (IR) and serum resistin level were also analyzed along with metabolic risk factors. Of 192 abdominal obese women, 147 (76.56%) were found to have mutant CC+GC (p = 0.001) genotype and allele frequency (p = 0.001), which was significantly higher 45 (23.44%) than non-obese and their respective wild type. The mutant genotype (CC+GC) of IL-6 gene was found to be associated significantly with high triglyceride (p = 0.025) and resistin level (p < 0.001), when compared with respective wild genotype (GG) in obese women. Non-obese women with no signs of metabolic risk factors were found to have significantly low level of serum resistin and IR in comparison to obese women having genetic polymorphism for IL-6 G-174C gene. Study suggests that IL-6 G-174C gene is one among the susceptibility loci for metabolic syndrome in North Indian women. Genotype for this polymorphism may prove informative for prediction of genetic risk for metabolic syndrome. Further, high level of serum resistin molecules may be targeted to correlate with metabolic syndrome risk factors and could be used as early prediction marker.

Cisplatin combination chemotherapy induces oxidative stress in advance non small cell lung cancer patients.

BACKGROUND: This study determine oxidative stress and survival prospectively in advanced stage non small cell lung cancer (NSCLC) patients following cisplatin based combination chemotherapy. MATERIALS AND METHODS: The oxidative stress levels (LPO, NO, GSH and SOD) of 144 control subjects and 203 advanced stage (IIIA/IIIB/IV) newly diagnosed NSCLC patients were assessed at pre-treatment (day '0'), and after the 3rd and 6th cycles of chemotherapy. Groups were compared by repeated measures ANOVA while comparison of survival curves was conduced by Kaplan-Meier methods. RESULTS: The pre-treatment mean levels of LPO and NO in patients were significantly (P<0.01) higher while GSH and SOD were significantly (P<0.01) lower as compared to control. The oxidative stress was elevated more significantly (P<0.01) after the chemotherapy and was more evident in higher stage than lower stage patients. The two year overall survival (%) of stage IV patients was significantly lower (P<0.05) as compared to stage III A and III B. The proportional mortality was also maximal in stage IV patients (37.0%) followed by stage III B (31.7%) and III A (20.0%). CONCLUSION: Cisplatin based combination chemotherapy induces oxidative stress in NSCLC patients.

Expression profiling of G2/M phase regulatory proteins in normal, premalignant and malignant uterine cervix and their correlation with survival of patients.

BACKGROUND: Cell regulatory G2/M phase proteins are the key regulators of mitosis and have been reported with abnormal expressions in various malignancies. AIM: To determine the expressions of these proteins in neoplastic uterine cervix tissue. MATERIALS AND METHODS: This study evaluates the G2/M phase regulatory protein expression of Cyclin B1, Aurora-B, Pololike kinase 1 (PLK1) and LIM kinase1 (LIMK1) in tissues of 25 normal (control), 16 dysplastic (dysplasia) and 34 neoplastic (cancer) patients of uterine cervix. The expressions of different proteins were obtained by using Western Blot technique. STATISTICAL ANALYSIS: One way analysis of variance (ANOVA), Pearson correlation, Kaplan-Meier and other tests are used for analysis. RESULTS AND CONCLUSION: The level of expression of LIMK1 in cervical cancer patients was found to be significantly higher (P < 0.01) than both the controls and dysplasia. The expression of Aurora B and PLK1 in cervical cancer patients was also found to be significantly higher ( P < 0.05) than controls but it did not differ with dysplasia. However, the expression of Cyclin B1 was similar among cervical cancer patients, dysplasia and controls ( P> 0.05). The expression of all the above proteins showed significant ( P < 0.01) and inverse relation with the survival of cancer patients. Among the selected candidate proteins, it was LIMK1 that showed the most positive correlation with the aggressiveness of the disease and negative correlation (r= -0.64; P < 0.01) with the survival of patients.

A randomized double blind placebo controlled clinical evaluation of extract of Andrographis paniculata (KalmCold) in patients with uncomplicated upper respiratory tract infection.

A randomized, double blind placebo controlled clinical study was conducted to evaluate the efficacy of KalmCold, an extract of Andrographis paniculata, in patients with uncomplicated upper respiratory tract infection (URTI). The assessment involved quantification of symptom scores by Visual Analogue Scale. Nine self evaluated symptoms of cough, expectoration, nasal discharge, headache, fever, sore throat, earache, malaise/fatigue and sleep disturbance were scored. A total of 223 patients of both sexes were randomized in two groups which received either KalmCold (200 mg/day) or placebo in a double blind manner. In both the treatments, mean scores of all symptoms showed a decreasing trend from day 1 to day 3 but from day 3 to day 5 most of the symptoms in placebo treated group either remained unchanged (cough, headache and earache) or got aggravated (sore throat and sleep disturbance) whereas in KalmCold treated group all symptoms showed a decreasing trend. Within groups, mean scores of symptoms in both the groups decreased significantly (p < or = 0.05) from day 1 to day 3 and day 5 while from day 3 to day 5 all symptoms except expectoration in placebo group did not improve significantly whereas in KalmCold treated group all symptoms improved significantly (p < or = 0.05) except earache. Comparing mean between both groups, all symptoms at day 1 and day 3 were found to be the same while at day 5 all symptoms except earache in KalmCold treated group improved significantly (p < or = 0.05) than placebo group. Similarly, within groups, overall scores of all symptoms in both the groups decreased significantly (p < or = 0.05) from day 1 to day 3 and day 5 while from day 3 to day 5 placebo group did not improve significantly whereas KalmCold treated group showed significant improvement (p < or = 0.05). On between groups analysis, KalmCold group showed significant reduction (p < or = 0.05) in overall symptom scores as compared to placebo group. In both placebo and KalmCold treated groups, there were only a few minor adverse effects with no significant difference in occurrence (Z = 0.63; p > 0.05). The comparison of overall efficacy of KalmCold over placebo was found to be significant (p < or = 0.05) and it was 2.1 times (52.7%) higher than placebo. The findings of this study revealed that KalmCold was effective in reducing symptoms of upper respiratory tract infection.