Impact of THIONIL® on reducing water and soil micro-pollutant and enhancement of growth, survival, anti-vibrio, immunity and health indices of Litopenaeus vannamei in pond trial.

There is an urgent requirement for the improvement of the white leg shrimp, Litopenaeus vannamei; health-related indices; and immunity due to emerging diseases. Recently, probiotics have been playing an important role in L. vannamei health management. Therefore, the current pond trial was to evaluate the probiotic proficiency of commercial probiotic products of THIONIL (THIO) on the enhancement of the water, soil, growth, digestibility, survival, immune-related indices, and susceptibility of L. vannamei to infection. The study was carried out in the major shrimp culturing regions of Kavali, Nellore (Andhra Pradesh), and Ponneri (Tamil Nadu), India. Six groups (lacks/ha) of the experimental L. vannamei were allocated, including a control group (THIO 0%-untreated) and groups containing 2%, 4%, 6%, 8%, and 10% of THIO that were encapsulated with commercial feed (CP Aqua). Bioassays were performed on PLs/ shrimp at various days interval of 0, 5, 25, 50, 100, and 123th to assess productivity, anti-vibrio activity, and digestive enzyme for digestibility, histological and immunological indices, and cytotoxicity in Artemia nauplii. Significant differences were observed in the increased growth (35.71 ± 3.24 g/shrimp) and digestive parameters in 10% THIO-fed shrimp. Although in contrast to the control group, the other THIO-fed prawn groups also displayed appreciable development. The findings showed that, in comparison to the control, the gill, hepatopancreas, and stomach had reduced tissue damage with 10% THIO. Furthermore, Vibrio parahaemolyticus (0.008 × 104 cfu/g) and Vibrio harveyi (0.051 × 105 cfu/g) (vibriosis) were potentially resistant to the 10% THIO-fed group. In addition, THIO-fed prawns (10%) showed significant improvements in immune-related expresses (proPO, SOD, and SOA) in comparison to the control. In conclusion, the findings showed that the THIO treatment prawns significantly improved the quality of their water (pH, ammonia, nitrogen dioxide, hydrogen sulfide, and DO) and soil (Pb, Cr, Hg, Mg, Cu, Fe, and Ni), increased and demonstrated protection against vibrio infections.

Biochemical and molecular anticancer approaches for Boerhaavia diffusa root extracts in oral cancer.

Background: Boerhaavia diffusa is a medicinal herb with anti-inflammatory, antiproliferative, anticancer, and immunomodulatory properties, found across India. Aim and Objectives: The present study is designed to investigate the therapeutic potential for B. diffusa root extracts in oral cancer cell line. Materials and Methods: The aqueous and methanolic extracts of B. diffusa were prepared using Soxhlet apparatus. In order to determine the phytochemical constituents of B. diffusa, the extracts were subjected to gas chromatography-mass spectrometry analysis. The antioxidant potential of B. diffusa extracts was assessed by 2,2-Diphenyl-picrylhydrazyl, ferric ion-reducing antioxidant power, catalase and peroxidase assays. The effective concentration of B. diffusa root on cell viability was analyzed by [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The ability of B. diffusa root extracts to modify the cell-cycle phases was performed by FACS analysis. The apoptotic inducing potential of B. diffusa in oral cancer cells was confirmed by acridine orange-ethidium bromide and 4',6-diamidino-2-phenylindole staining. The protein profile of apoptotic processes was validated by the Western blot analysis; docking studies were also performed. Results: We observed that antioxidant activity was higher in B. diffusa methanolic extract compared with aqueous extract. The results showed that the methanolic and aqueous extracts of B. diffusa exhibited significant cytotoxic effect with IC50 value of 36 µg/ml and 30 µg/ml, respectively. The apoptotic DNA fragmentation and the apoptotic inducing potential in KB oral cancer cell line were higher for the methanolic extract compared with the aqueous extract. These results were also confirmed by in-silico analysis. Conclusion: The results indicate that extracts obtained from the roots of B. diffusa inhibit the progression of oral cancer. These compounds of pharmacological importance can be either used alone or in combination with other drugs to treat oral cancer.

Identification of promoter PcadR, in silico characterization of cadmium resistant gene cadR and molecular cloning of promoter PcadR from Pseudomonas aeruginosa BC15.

Cadmium (Cd) remediation in Pseudomonas aeruginosa is achieved through the function of two vital genes, cadA and cadR, that code for P-type ATPase (CadA) and transcription regulatory protein (CadR), respectively. Although numerous studies are available on these metal-sensing and regulatory proteins, the promoter of these genes, metal sensing and binding ability, are poorly understood. The present work is aimed at the characterization of the CadR protein, identification of the PcadR promoter and protein-promoter-metal binding affinity using bioinformatics and to validate the results by cloning the PcadR promoter in Escherichia coli DH5α. The promoter regions and its curvature were identified and analysed using PePPER software (University of Groningen, The Netherland) and the Bendit program (Version: v.1.0), respectively. Using Phyre, the three-dimensional structure of CadR was modelled, and the structure was validated by Ramachandran plots. The DNA-binding domain was present in the N-terminal region of CadR. A dimeric interface was observed in helix-turn-helix and metal ion-binding sites at the C-terminal. Docking studies showed higher affinity of Cd to both CadR (Atomic contact energy = -15.04 kcal/Mol) and PcadR (Atomic contact energy = -40.18 kcal/Mol) when compared to other metal ions. CadR with PcadR showed the highest binding affinity (Atomic contact energy= -250.40 kcal/Mol) when compared with PcadA. In vitro studies using green fluorescent protein tagged with PcadR (gfp-PcadR) cloned in E. coli-expressed gfp protein in a concentration-dependent manner upon Cd exposure. Based on our in silico studies and in vitro molecular cloning analysis, we conclude that PcadR and CadR are active only in the presence of Cd. The CadR protein has the highest binding affinity with PcadR. As it became apparent that the cadR gene regulates the PcadR activity in the presence of Cd with high specificity, and the cadR and PcadR can be used as a biological tool for development of a microbial biosensor.

Oral administration of oxalate-enriched spinach extract as an improved methodology for the induction of dietary hyperoxaluric nephrocalcinosis in experimental rats.

Experimental induction of hyperoxaluria by ethylene glycol (EG) administration is disapproved as it causes metabolic acidosis while the oral administration of chemically synthesized potassium oxalate (KOx) diet does not mimic our natural system. Since existing models comprise limitations, this study is aimed to develop an improved model for the induction of dietary hyperoxaluria, and nephrocalcinosis in experimental rats by administration of naturally available oxalate rich diet. Male albino Wistar rats were divided into five groups. Group I, control; group II rats received 0.75% EG, group III rats fed with 5% KOx diet and group IV and V rats were administered with spinach extract of 250 and 500 mg soluble oxalate/day respectively, for 28 d. Urine and serum biochemistry were analyzed. After the experimental period, rats were sacrificed, liver and kidney tissue homogenates were used for antioxidant and lipid peroxidation assay. Relative change in expression of kidney injury molecule-1 (KIM-1) and crystal modulators genes in kidney tissues were evaluated. Tissue damage was assessed by histology studies of liver and kidney. Experimental group rats developed hyperoxaluria and crystalluria. Urine parameters, serum biochemistry, antioxidant profile, lipid peroxidation levels and gene expression analysis of experimental group II and III rats reflected acute kidney damage compared to group V rats. Histopathology results showed moderate hyperplasia in liver and severe interstitial inflammation in kidneys of group II and III than group V rats. Ingestion of naturally available oxalate enriched spinach extract successfully induced dietary hyperoxaluria and nephrocalcinosis in rats with minimal kidney damage.