Adrenergic and muscarinic receptor subtypes functional regulation during diabetogenesis: Effect of curcumin and vitamin D3 pre-treatment

In the present study, the initial phase was directed to confirm the effects of curcumin and vitamin D3 in preventing or delaying diabetes onset by studying the blood glucose and insulin levels in the pre-treated and diabetic groups. Behavioural studies were conducted to evaluate the cognitive and motor function in experimental rats. The major focus of the study was to understand the cellular and neuronal mechanisms that ensure the prophylactic capability of curcumin and vitamin D3. To elucidate the mechanisms involved in conferring the antidiabetogenesis effect, we examined the DNA and protein profiles using radioactive incorporation studies for DNA synthesis, DNA methylation and protein synthesis. Furthermore the gene expression studies of Akt-1, Pax, Pdx-1, Neuro D1, insulin like growth factor-1 and NF-κB were done to monitor pancreatic beta cell proliferation and differentiation. The antioxidant and antiapoptotic actions of curcumin and vitamin D3 were examined by studying the expression of antioxidant enzymes - SOD and GPx, and apoptotic mediators like Bax, caspase 3, caspase 8 and TNF-α. In order to understand the signalling pathways involved in curcumin and vitamin D3 action, the second messengers, cAMP, cGMP and IP3 were studied along with the expression of vitamin D receptor in the pancreas. The neuronal regulation of pancreatic beta cell maintenance, proliferation and insulin release was studied by assessing the adrenergic and muscarinic receptor functional regulation in the pancreas, brain stem, hippocampus and hypothalamus. The receptor number and binding affinity of total muscarinic, muscarinic M1, muscarinic M3, total adrenergic, α adrenergic and β adrenergic receptor subtypes were studied in pancreas, brain stem and hippocampus of experimental rats. The mRNA expression of muscarinic and adrenergic receptor subtypes were determined using Real Time PCR. Immunohistochemistry studies using confocal microscope were carried out to confirm receptor density and gene expression results. Cell signalling alterations in the pancreas and brain regions associated with diabetogenesis and antidiabetogenesis were assessed by examining the gene expression profiles of vitamin D receptor, CREB, phospholipase C, insulin receptor and GLUT. This study will establish the anti-diabetogenesis activity of curcumin and vitamin D3 pre-treatment and will attempt to understand the cellular, molecular and neuronal control mechanism in the onset of diabetes.Administration of MLD-STZ to curcumin and vitamin D3 pre-treated rats induced only an incidental prediabetic condition. Curcumin and vitamin D3 pretreated groups injected with MLD-STZ exhibited improved circulating insulin levels and behavioural responses when compared to MLD-STZ induced diabetic group. Activation of beta cell compensatory response induces an increase in pancreatic insulin output and beta cell mass expansion in the pre-treated group. Cell signalling proteins that regulate pancreatic beta cell survival, insulin release, proliferation and differentiation showed a significant increase in curcumin and vitamin D3 pre-treated rats. Marked decline in α2 adrenergic receptor function in pancreas helps to relent sympathetic inhibition of insulin release. Neuronal stimulation of hyperglycemia induced beta cell compensatory response is mediated by escalated signalling through β adrenergic, muscarinic M1 and M3 receptors. Pre-treatment mediated functional regulation of adrenergic and cholinergic receptors, key cell signalling proteins and second messengers improves pancreatic glucose sensing, insulin gene expression, insulin secretion, cell survival and beta cell mass expansion in pancreas. Curcumin and vitamin D3 pre-treatment induced modulation of adrenergic and cholinergic signalling in brain stem, hippocampus and hypothalamus promotes insulin secretion, beta cell compensatory response, insulin sensitivity and energy balance to resist diabetogenesis. Pre-treatment improved second messenger levels and the gene expression of intracellular signalling molecules in brain stem, hippocampus and hypothalamus, to retain a functional neuronal response to hyperglycemia. Curcumin and vitamin D3 protect pancreas and brain regions from oxidative stress by their indigenous antioxidant properties and by their ability to stimulate cellular free radical defence system. The present study demonstrates the role of adrenergic and muscarinic receptor subtypes functional regulation in curcumin and vitamin D3 mediated anti-diabetogenesis. This will have immense clinical significance in developing effective strategies to delay or prevent the onset of diabetes.

Process Optimization for Mass Production of Marine Yeast Candida sp. S 27 and its Nutritional Characterization

The main source of protein for human and animal consumption is from the agricultural sector, where the production is vulnerable to diseases, fluctuations in climatic conditions and deteriorating hydrological conditions due to water pollution. Therefore Single Cell Protein (SCP) production has evolved as an excellent alternative. Among all sources of microbial protein, yeast has attained global acceptability and has been preferred for SCP production. The screening and evaluation of nutritional and other culture variables of microorganisms are very important in the development of a bioprocess for SCP production. The application of statistical experimental design in bioprocess development can result in improved product yields, reduced process variability, closer confirmation of the output response to target requirements and reduced development time and overall cost.The present work was undertaken to develop a bioprocess technology for the mass production of a marine yeast, Candida sp.S27. Yeasts isolated from the offshore waters of the South west coast of India and maintained in the Microbiology Laboratory were subjected to various tests for the selection of a potent strain for biomass production. The selected marine yeast was identified based on ITS sequencing. Biochemical/nutritional characterization of Candida sp.S27 was carried out. Using Response Surface Methodology (RSM) the process parameters (pH, temperature and salinity) were optimized. For mass production of yeast biomass, a chemically defined medium (Barnett and Ingram, 1955) and a crude medium (Molasses-Yeast extract) were optimized using RSM. Scale up of biomass production was done in a Bench top Fermenter using these two optimized media. Comparative efficacy of the defined and crude media were estimated besides nutritional evaluation of the biomass developed using these two optimized media.

GABA and 5-HT chitosan nanoparticles enhanced liver cell proliferation and neuronal survival in partially hepatectomised rats: GABAB and 5-HT2A receptors functional

Nanoparticulate drug delivery systems provide wide opportunities for solving problems associated with drug stability or disease states and create great expectations in the area of drug delivery (Bosselmann & Williams, 2012). Nanotechnology, in a simple way, explains the technology that deals with one billionth of a meter scale (Ochekpe, et al., 2009). Fewer side effects, poor bioavailability, absorption at intestine, solubility, specific delivery to site of action with good pharmacological efficiency, slow release, degradation of drug and effective therapeutic outcome, are the major challenges faced by most of the drug delivery systems. To a great extent, biopolymer coated drug delivery systems coupled with nanotechnology alleviate the major drawbacks of the common delivery methods. Chitosan, deacetylated chitin, is a copolymer of β-(1, 4) linked glucosamine (deacetylated unit) and N- acetyl glucosamine (acetylated unit) (Radhakumary et al., 2005). Chitosan is biodegradable, non-toxic and bio compatible. Owing to the removal of acetyl moieties that are present in the amine functional groups of chitin, chitosan is readily soluble in aqueous acidic solution. The solubilisation occurs through the protonation of amino groups on the C-2 position of D-glucosamine residues whereby polysaccharide is converted into polycation in acidic media. Chitosan interacts with many active compounds due to the presence of amine group in it. The presence of this active amine group in chitosan was exploited for the interaction with the active molecules in the present study. Nanoparticles of chitosan coupled drugs are utilized for drug delivery in eye, brain, liver, cancer tissues, treatment of spinal cord injury and infections (Sharma et al., 2007; Li, et a., 2009; Paolicelli et al., 2009; Cho et al., 2010). To deliver drugs directly to the intended site of action and to improve pharmacological efficiency by minimizing undesired side effects elsewhere in the body and decrease the long-term use of many drugs, polymeric drug delivery systems can be used (Thatte et al., 2005).

Numerical Simulation of Two-Part Underwater Towing System

The motion instability is an important issue that occurs during the operation of towed underwater vehicles (TUV), which considerably affects the accuracy of high precision acoustic instrumentations housed inside the same. Out of the various parameters responsible for this, the disturbances from the tow-ship are the most significant one. The present study focus on the motion dynamics of an underwater towing system with ship induced disturbances as the input. The study focus on an innovative system called two-part towing. The methodology involves numerical modeling of the tow system, which consists of modeling of the tow-cables and vehicles formulation. Previous study in this direction used a segmental approach for the modeling of the cable. Even though, the model was successful in predicting the heave response of the tow-body, instabilities were observed in the numerical solution. The present study devises a simple approach called lumped mass spring model (LMSM) for the cable formulation. In this work, the traditional LMSM has been modified in two ways. First, by implementing advanced time integration procedures and secondly, use of a modified beam model which uses only translational degrees of freedoms for solving beam equation. A number of time integration procedures, such as Euler, Houbolt, Newmark and HHT-α were implemented in the traditional LMSM and the strength and weakness of each scheme were numerically estimated. In most of the previous studies, hydrodynamic forces acting on the tow-system such as drag and lift etc. are approximated as analytical expression of velocities. This approach restricts these models to use simple cylindrical shaped towed bodies and may not be applicable modern tow systems which are diversed in shape and complexity. Hence, this particular study, hydrodynamic parameters such as drag and lift of the tow-system are estimated using CFD techniques. To achieve this, a RANS based CFD code has been developed. Further, a new convection interpolation scheme for CFD simulation, called BNCUS, which is blend of cell based and node based formulation, was proposed in the study and numerically tested. To account for the fact that simulation takes considerable time in solving fluid dynamic equations, a dedicated parallel computing setup has been developed. Two types of computational parallelisms are explored in the current study, viz; the model for shared memory processors and distributed memory processors. In the present study, shared memory model was used for structural dynamic analysis of towing system, distributed memory one was devised in solving fluid dynamic equations.

Alkaline protease from a non-toxigenic mangrove isolate of Vibrio sp. V26 with potential application in animal cell culture

Vibrio sp. V26 isolated from mangrove sediment showed 98 % similarity to 16S rRNA gene of Vibrio cholerae, V. mimicus, V. albensis and uncultured clones of Vibrio. Phenotypically also it resembled both V. cholerae and V. mimicus.Serogrouping, virulence associated gene profiling, hydrophobicity, and adherence pattern clearly pointed towards the non—toxigenic nature of Vibrio sp. V26. Purification and characterization of the enzyme revealed that it was moderately thermoactive, nonhemagglutinating alkaline metalloprotease with a molecular mass of 32 kDa. The application of alkaline protease from Vibrio sp. V26 (APV26) in sub culturing cell lines (HEp-2, HeLa and RTG-2) and dissociation of animal tissue (chick embryo) for primary cell culture were investigated. The time required for dissociation of cells as well as the viable cell yield obtained by while administeringAPV26 and trypsin were compared. Investigations revealed that the alkaline protease of Vibrio sp. V26 has the potential to be used in animal cell culture for subculturing cell lines and dissociation of animal tissue for the development of primary cell cultures, which has not been reported earlier among metalloproteases of Vibrios.