Unraveling the role of intra-cellular metabolites in the lactic acid production by novel Bacillus amyloliquefaciens using sugarcane molasses as a substratum.

Lactic acid is a versatile, multi-functional organic monomer in various industries, creating worldwide demand. High titer lactic acid production was achieved by novel Bacillus amyloliquefaciens J2V2AA through sugarcane molasses fermentation up to 178 mg mL-1. A metabolomics approach such as combined GC-MS and LC-MS was applied to elucidate the involvement of key metabolites in lactic acid production. The results revealed the participation of 58 known intra-cellular metabolites at various pathways in lactic acid production. Twenty-eight highly up-regulated and down-regulated metabolites were analyzed, and a schematic diagram of a possible lactic acid production pathway was proposed. The produced lactic acid was analyzed through FTIR, UV-Spectrum, and HPLC analysis.

Progressive Spreading of DNA Methylation in the GSTP1 Promoter CpG Island across Transitions from Precursors to Invasive Prostate Cancer.

Glutathione S-transferase pi 1 (GSTP1) is lowly expressed in normal prostate luminal cells and becomes induced in most proliferative inflammatory atrophy (PIA) lesions. GSTP1 becomes silenced in prostatic intraepithelial neoplasia (PIN) and prostate adenocarcinoma (CaP) via cytosine-phospho-guanine (CpG) island promoter hypermethylation. However, GSTP1 methylation patterns in PIA and PIN, and their relationship to patterns in CaP are poorly understood. We used bisulfite genomic sequencing to examine patterns of GSTP1 promoter CpG island methylation in laser capture microdissected benign, PIA, PIN, and CaP regions from 32 subjects that underwent radical prostatectomy. We analyzed 908 sequence clones across 24 normal epithelium, 37 PIA, 18 PIN, and 23 CaP regions, allowing assessment of 34,863 CpG sites with allelic phasing. Normal and PIA lesions were mostly unmethylated with 0.52 and 1.3% of total CpG sites methylated, respectively. PIN and CaP lesions had greater methylation with 24% and 51% of total CpG sites methylated, respectively. The degree of GSTP1 methylation showed progression from PIA << PIN < CaP. PIN lesions showed more partial methylation compared with CaP lesions. Partially methylated lesions were enriched for methylation changes at AP1 and SP1 transcription factor binding sites. These results demonstrate that methylation density in the GSTP1 CpG island in PIN was intermediate relative to that in normal prostate epithelium/PIA and CaP lesions. These results are consistent with gradual spreading of DNA methylation centered at the SP1/AP1 transcription factor binding sites in precursor lesions, with subsequent spreading of methylation across the entire CpG island in transition to CaP. PREVENTION RELEVANCE: DNA hypermethylation at the GSTP1 promoter progressively spreads from being unmethylated in normal prostate to intermediate levels in precursor lesions to extensive methylation in cancer. This molecular progression of GSTP1 promoter methylation patterns in early prostate carcinogenesis could be useful for identification and interception of prostate cancer precursors.

Optimization of sodium alginate-galactoxyloglucan blended hydrogel beads through ionotropic gelation method.

Hydrogels are 3D crosslinking networks of hydrophilic biopolymers which can able to absorb and retain large amount of water. In this present study, the Sodium alginate (SA)- Galactoxyloglucan (GXG) blended hydrogel beads were prepared and optimized through two level optimization steps. Alginate and xyloglucan are the cell wall polysaccharides biopolymers obtained from the plant sources, Sargassum sp. and Tamarindus indica L. respectively. The extracted biopolymers were confirmed and characterized by UV-Spectroscopy, FT-IR, NMR and TGA analysis. Based on the hydrophilicity, non-toxicity and biocompatibility, SA-GXG hydrogel were prepared and optimized through two-level optimization steps. The optimized hydrogel bead formulation was characterized through FT-IR, TGA and SEM analysis. From the obtained result, it is found that the polymeric formulation GXG (2 % w/v)-SA (1.5 % w/v), cross-linker (CaCl2) concentration at 0.1 M and the cross-linking time at 15 Min showed significant swelling index. The optimized hydrogel beads are porous and show good swelling capacity and thermal stability. The optimized protocol of hydrogel beads may be useful in designing hydrogel beads for specific applications in agricultural, Biomedical and remediation sectors.

Pervasive promoter hypermethylation of silenced TERT alleles in human cancers.

BACKGROUND: In cancers, maintenance of telomeres often occurs through activation of the catalytic subunit of telomerase, encoded by TERT. Yet, most cancers show only modest levels of TERT gene expression, even in the context of activating hotspot promoter mutations (C228T and C250T). The role of epigenetic mechanisms, including DNA methylation, in regulating TERT gene expression in cancer cells is as yet not fully understood. METHODS: Here, we have carried out the most comprehensive characterization to date of TERT promoter methylation using ultra-deep bisulfite sequencing spanning the CpG island surrounding the core TERT promoter in 96 different human cell lines, including primary, immortalized and cancer cell types, as well as in control and reference samples. RESULTS: In general, we observed that immortalized and cancer cell lines were hypermethylated in a region upstream of the recurrent C228T and C250T TERT promoter mutations, while non-malignant primary cells were comparatively hypomethylated in this region. However, at the allele-level, we generally found that hypermethylation of promoter sequences in cancer cells is associated with repressed expression, and the remaining unmethylated alleles marked with open chromatin are largely responsible for the observed TERT expression in cancer cells. CONCLUSIONS: Our findings suggest that hypermethylation of the TERT promoter alleles signals transcriptional repression of those alleles, leading to attenuation of TERT activation in cancer cells. This type of fine tuning of TERT expression may account for the modest activation of TERT expression in most cancers.

Ameliorative potential of Solanum trilobatum leaf extract and fractions on lipid profile and oxidative stress in experimental diabetes.

Solanum trilobatum Linn is a medicinal plants used in India from many centuries to cure various diseases. The purpose of the study is to evaluate the ameliorative potential of the ethanolic leaf extract and fractions of Solanum trilobatum (St) against alloxan induced diabetic rats. Solanum trilobatum leaf extract and fractions were administered orally at two different concentration (100-200mg/kg body weight) to alloxan induced diabetic rats. The antidiabetic efficacy was validated through various biochemical parameters, enzyme assays, renal markers and antioxidant properties were also determined. The phytochemical analyses of St leaves were done by estimating their Chlorophyll, carotenoids, total sugar, protein, amino acid and minerals contents. The results revealed that the leaf extracts and fractions of St were efficient anti hyperglycemic agents and posses potent nephro-protective activity. However, the extracts of St leaves at a concentration of 200mg/kg bw exhibit higher efficacy in all tested concentrations. From the result it can be concluded that the leaf extracts of St can be a potential candidate in treating the hyperglycemic conditions and justifies its use in ethano medicine and can be exploits in the management of diabetes.

The effects of over expressing aquaporins on the cryopreservation of hepatocytes.

During cryopreservation, aquaporins are critical in regulating water transport across cellular membranes and preventing osmotic damages. Hepatocytes express aquaporin (AQP) 0, 8, 9, 11, and 12; this study investigates whether increasing the localization of AQP8 on the cellular membrane would improve cell viability by increasing water transport during cryopreservation. Primary rat hepatocytes were cultured and treated with dibutyryl cAMP (Bt(2)cAMP) or glucagon to increase the expression of AQP8 at the cellular membrane via translocation. This phenomenon is verified through two experiments - confocal immunofluorescence microscopy and cell shrinkage analysis. The immunofluorescence results showed increase in AQP8 on the cellular membrane of treated cells, and cell shrinkage analysis showed an increase in water transport of treated cells compared to controls. Primary rat hepatocytes were treated with Bt(2)cAMP or glucagon and cryopreserved using standard protocols in a controlled rate freezer. This resulted in a significant increase in the cell viability on warming. These results indicate that Bt(2)cAMP or glucagon treated hepatocytes had increased expression of aquaporin in the cellular membrane, increased water transport during cryopreservation, and increased post-thaw viability.

Near infra-red laser mediated photothermal and antitumor efficacy of doxorubicin conjugated gold nanorods with reduced cardiotoxicity in swiss albino mice.

Development of a multifunctional drug delivering system without side effects and compromising its therapeutic efficacy is a major concern in anticancer research. Recently, we have developed and demonstrated doxorubicin conjugated gold nanorod (DOX@PSS-GNR) as a sustained drug delivery vehicle. Here, we investigate the biodistribution, antitumor and photothermal efficacy of DOX@PSS-GNR along with its potential impact on cardiotoxicity in in vivo. The studies revealed that the accumulation of Free DOX in myocardium was 4-fold reduced in DOX@PSS-GNR animals, which further minimizes its cardiotoxicity by decreasing cardiac injury via preservation of cardiac markers. Further, DOX@PSS-GNR exhibits effective antitumor efficacy against Dalton lymphoma ascites (DLA) as evidenced by cell cycle analysis, apoptotic signals and reduced tumor volume and weight. In addition, DOX@PSS-GNR exhibits higher photothermal response and dominates DLA growth upon 0.1 W/cm(2) laser irradiation. In conclusion, multifunctional DOX@PSS-GNR with improved therapeutic index and reduced cardiotoxicity represents a promising candidate for cancer treatment. FROM THE CLINICAL EDITOR: Doxorubicin is a widely used agent for cancer therapy. However, the side effects are still significant, despite the development of liposomal formulation. In this study, the authors investigated the use of doxorubicin conjugated gold nanorods (DOX@PSS-GNR) in terms of biodistribution, antitumor activity and systemic side effects. The much reduced cardiotoxicity of the new delivery system should provide an improved agent for future clinical use.

An update on prodrugs from natural products.

A natural prodrug is a chemical compound or substance obtained from plants, microorganism, animal and marine sources. Natural products are small molecule source for Food and Drug Administration approved drugs and major sources for drug discovery. Most of the drugs for different ailment diseases undergo first pass metabolism, resulting in drug inactivation and the generation of toxic metabolites in body. Enormous numbers of prodrugs naturally present in plants, microorganism, animal and marine sources and those prodrugs undergoes chemical reaction to form non-toxic compounds. This review summarizes the list of prodrugs naturally present in the natural product.

Chemopreventive effect of montelukast in n-nitroso n-methyl urea induced mammary carcinogenesis in female Sprague-Dawley rats.

OBJECTIVE: The objective of the study was to evaluate the chemopreventive effect of montelukast sodium; selective reversible cysteinyl leukotriene D4-receptor antagonist in N-nitroso N-methyl urea (NMU) induced mammary carcinogenesis in virgin female Sprague-Dawley rats. MATERIALS AND METHODS: Thirty rats were divided into five groups (normal control, disease control, montelukast1 mg/kg, montelukast10 mg/kg, tamoxifen10 mg/kg) of six animals each. The drug was administered in two doses,1 mg/kg,and 10 mg/kg orally and compared with the standard drug tamoxifen (10 mg/kg)p.o. RESULTS: Montelukast sodium 1 mg/kg,10 mg/kg, and tamoxifen10 mg/kg decreased the tumor incidences by 50%,66.67%, and 83.33% and the total number of tumors in group by 41.67%, 58.33% and 91.67% respectively, when compared to the disease control. Montelukast sodium 1 mg/kg,10 mg/kg,and tamoxifen10 mg/kg decreased the average tumor burden by 86.41%,94.8% and 95.97%and average tumor volume by 89.52%, 95.84%, and 95.4%respectively, when compared to disease control group. CONCLUSION: The results revealed that montelukast sodium prevent the mammary carcinogenesis and confirms the role of cysteinyl leukotriene D4-receptor in mammary gland neoplasia.

Effect of silymarin on streptozotocin-nicotinamide-induced type 2 diabetic nephropathy in rats.

INTRODUCTION: Diabetic nephropathy is the major cause of end-stage renal disease worldwide. Silymarin is a flavonoid mixture obtained from Silybum marianum. Various preclinical and clinical studies have revealed that it has antidiabetic activity. The objective of this study was to evaluate the effect of silymarin on type 2 diabetic nephropathy in rats. MATERIALS AND METHODS: Non-insulin-dependent diabetes mellitus was induced in overnight-fasted male albino Wistar rats by an intramuscular injection of streptozotocin and nicotinamide. Eighteen rats with diabetic nephropathy and 6 rats without induced nephropathy were divided into 4 groups, each containing 6 animals. Group 1 was the normal control and group 2 was the DM control. Groups 3 and 4 were rats with diabetic nephropathy that received 60 mg/kg and 120 mg/kg of silymarin for 60 days. RESULTS: At the end of the study period, the diabetic control group had significantly higher blood glucose, glycosylated hemoglobin, urine volume, serum uric acid, and urine albumin levels when compared to the normal control group. Silymarin-treated groups showed significantly lower levels of blood glucose, glycosylated hemoglobin, urine volume, serum creatinine, serum uric acid, and urine albumin, when compared to the diabetic control group. Histopathological studies reports strongly supported the protective effect of silymarin. CONCLUSIONS: These findings suggest that silymarin has protective effects for kidneys affected by type 2 diabetes mellitus. If the safety and efficacy is confirmed in humans, silymarin will be a good medication to prevent nephropathy-induced premature death in diabetic patients.

Molecular alterations in colitis-associated colorectal neoplasia: study from a low prevalence area using magnifying chromo colonoscopy.

BACKGROUND AND AIM: Longstanding ulcerative colitis (UC) predisposes to colorectal cancer (CRC). To understand the molecular pathogenesis of colitis-associated colorectal neoplasia (UC-CRN), we studied the frequency of microsatellite instability (MSI) and mutations in p53, BRAF and KRAS genes in the tissues of patients with long standing UC with or without neoplasia and compared them with colitis patients without risk of neoplasia, and those with sporadic colorectal neoplasia (S-CRN) in an area with lower prevalence for either disease. METHODS: Biopsies were obtained during magnifying chromo colonoscopy or routine colonoscopy in consecutive UC patients with high risk (UC-HR) and low risk (UC-LR) of neoplasia, and those with S-CRN. MSI (NCI-Bethesda panel) and mutations in p53, KRAS and BRAF genes were analysed. RESULTS: Twenty-eight patients with UC-HR, 30 with UC-LR and 30 with S-CRN were included. Six (21.4%) of UC-HR had neoplasia (Progressors). MSI was not detected in the UC-CRN group as compared to 5 (16.7%) in the S-CRN group. p53 mutations occurred in 1 (3.3%) of UC-LR, increasing to 6 (27.3%, P<0.05) and 3 (50%, P<0.05) in the UC-HR subgroups without and with neoplasia respectively, as against 10 (33.3%) in sporadic neoplasia group. KRAS mutations were found only in the presence of neoplasia. None showed the BRAF mutation. CONCLUSIONS: In a population with a lower prevalence for UC and CRC, the molecular pathogenesis of colitis-associated colorectal neoplasia is comparable to that reported from areas with a higher prevalence of these diseases, MSI being an exception.

The role of salivary gland scintigraphy in detection of salivary gland dysfunction in type 2 diabetic patients.

OBJECTIVE: The aim of the study was to evaluate the salivary gland dysfunction in a patient with uncontrolled type 2 diabetes using salivary gland scintigraphy. MATERIALS AND METHODS: patients included in the study were 32 uncontrolled type 2 diabetic and 30 normal healthy individuals. Patients having any other systemic(or) nervous illness(or) taking medications that could affect the normal functioning of the salivary gland were excluded from the study. The salivary gland scintigraphy was performed, with radioactivity measured at 1(st), 20(th), and 40(th) minutes. Twenty minutes after the injection, vitamin C chewable tablet was given to stimulate the secretion and continued until the end of the study period (40min). The data were replayed and regions of interest were chosen over four salivary glands to obtain the uptake ratio (UR) and excretory ratio(ER) of the salivary glands. RESULT: The scintigraphic total URand ER in diabetic and control groups was compared. The values in these two categories showed decrease in both UR and ER in diabetic patients, when compared to control patients. CONCLUSION AND SIGNIFICANCE: The result of this study suggests that salivary gland scintigraphy plays a significant role in the evaluation of salivary gland dysfunction in type 2 diabetic patients.

Evaluation of phytoconstituents and anti-nephrotoxic and antioxidant activities of Monochoria vaginalis.

Monochoria vaginalis is an herbaceous medicinal plant used to treat, liver problems India. Acetaminophen is a commonly used analgesic and antipyretic agent which, at high doses, causes liver and kidney necrosis in man and animals. The aim of the present study is to evaluate phytoconstituents and investigate the nephroprotective & antioxidant activities of the ethanol extract of Monochoria vaginalis on acetaminophen induced toxicity in rats. Phytoconstituents like n-hexadecanoic acid, 3-methyl- acetate-1-butanol, 1,1,3-triethoxy- propane, Z,Z,Z-1,4,6,9 - nonadecatetraene, undecanoic acid, 3-trifluoroacetoxy penta decane and 4-ethyl-5-octyl-2,2-bis (trifluoromethyl) - cis-1,3-dioxalone were identified from ethanol extract of Monochoria vaginalis by using a gas chromatograph-mass spectrograph (GC MS). Biochemical studies show that there is an increase in the levels of serum urea and creatinine along with an increase in the body weight and reduction in the levels of uric acid in acetaminophen induced groups. These values are retrieved significantly by treatment with Monochoria vaginalis extracts at two different doses. The antioxidant studies reveal that the levels of renal SOD, CAT, GSH and GPx in the APAP treated animals are increased significantly along with a reduced MDA content in ethanol extract of Monochoria vaginalis treated groups. Apart from these, histopathological changes also reveal the protective nature of the Monochoria vaginalis extract against acetaminophen induced necrotic damage of renal tissues. In conclusion, these data suggest that the ethanol extract of Monochoria vaginalis can prevent renal damage from APAP induced nephrotoxicity in rats and it is likely to be mediated through active phytoconstituents and its antioxidant activities.

Evaluation of genetic alterations in inhabitants of a naturally high level background radiation and Kudankulam nuclear power project site in India.

Evaluation of genetic alterations in inhabitants of an area of Tamil Nadu, India, chronically exposed to high background radiation (HBRA), was the major purpose of the present study. A total of 216 samples (exposed inhabitants, 108; control subjects, 108) were selected based on the confirmation of radiation dose level using thermoluminescence dosimetry (TLD). After signing a consent form, volunteers provided blood samples (5 ml each) to establish cell cultures at 52 h. One hundred complete metaphase cells from each subject were evaluated for karyotyping. The frequencies of chromosomal alterations (CA) were found to be higher in the exposed groups and the aberrations predominately observed were of chromatid-type. Smoking was found to have considerable effect on the frequency of CA in exposed subjects. With the comet assay for DNA damage, a significant increase in comet tail frequency was also observed in exposed subjects compared to controls. At present there are no radioepidemiological data regarding the cytogenetic studies in these areas. Furthermore, the Kudankulam nuclear power plant nuclear power plant is being constructed in the same area. The study gives potentially important information on the general health effects due to radiation exposure and increases people's understanding of the hazardous nature of chronic low level natural radiation exposure. However, we may conclude that the HBRA by itself does not pose any significant risk of genetic damage as measured by conventional cytogenetic analysis.