Amelioration of ionizing radiation induced lipid peroxidation in mouse liver by Moringa oleifera Lam. leaf extract

Protective effect of Moringa oleifera leaf extract (MoLE) against radiation-induced lipid peroxidation has been investigated. Swiss albino mice, selected from an inbred colony, were administered with MoLE (300 mg/kg body wt) for 15 days before exposing to a single dose of 5 Gy 60Co-gamma radiation. After treatments, animals were necropsied at different post irradiation intervals (days 1, 7 and 15) and hepatic lipid peroxidation and reduced glutathione (GSH) contents were estimated to observe the relative changes due to irradiation and its possible amelioration by MoLE. It was observed that, MoLE treatment restored GSH in liver and prevented radiation induced augmentation in hepatic lipid peroxidation. Phytochemical analysis showed that MoLE possess various phytochemicals such as ascorbic acid, phenolics (catechin, epicatechin, ferulic acid, ellagic acid, myricetin) etc., which may play the key role in prevention of hepatic lipid peroxidation by scavenging radiation induced free radicals.     

LIBS Based Screening of Glycemic Elements of Ficus religiosa

The effectiveness of various trace element concentrations in medicinal plants in the cure of various diseases can be determined by their quantitative estimation. Elemental concentrations of aqueous extract of F. religiosa leaves were measured by Laser-induced breakdown spectroscopy (LIBS). LIBS is a very powerful and efficient analytical tool for determining elemental constitution. The present study deals with the LIBS-based validation of elements responsible for the glycemic potential of aqueous extract of F. religiosa leaves in streptozotocin-induced diabetic models. The significant decrease in blood glucose level and marked improvement in glucose tolerance test of diabetic models is correlated to the concentration of elements present in the extract as revealed by LIBS spectra. Elements such as Mg and Ca have been observed in the LIBS spectra of F. religiosa.     

Biochemical analysis of four species ofCassia L. pollen

The present paper describes the comparative biochemical studies in terms of quantitative analyses of carbohydrates, lipids, proteins, free amino acids, nucleic acids, minerals, ash and moisture as well as the identification of free amino acids of pollen of four species ofCassia L. (C. alata L,C. fistula L,C. occidentalis L andC. siamea Lam.). A significant variation in the chemical constituents was observed among the four species.C. occidentalis showed the highest levels of carbohydrate (15.15%) and protein (22.45%), andC. siamea had the lowest levels of carbohydrate (7.15%), lipid (6.2%) and protein (13.85%).C. alata andC. fistula showed intermediate results. However,C. alata showed the highest amount of free amino acids (3.8%) and the least of 1.42% was found inC. fistula. Thin layer chromatography (TLC) of free amino acids of the four species showed some homology in their amino acid content, of which proline, glutamic acid, methionine and phenyl-alanine were the most dominant. The level of nucleic acids and minerals was found to be comparatively low.C. siamea andC. alata showed an exceptionally high level of ash content (8.6 and 8.8%, respectively) while moisture content varied from 8 to 11%.     

Draft genome sequence of Arthrospira platensis C1 (PCC9438)

Arthrospira platensis is a cyanobacterium that is extensively cultivated outdoors on a large commercial scale for consumption as a food for humans and animals. It can be grown in monoculture under highly alkaline conditions, making it attractive for industrial production. Here we describe the complete genome sequence of A. platensis C1 strain and its annotation. The A. platensis C1 genome contains 6,089,210 bp including 6,108 protein-coding genes and 45 RNA genes, and no plasmids. The genome information has been used for further comparative analysis, particularly of metabolic pathways, photosynthetic efficiency and barriers to gene transfer.     

Expression, purification, and characterization of a bacterial GTP-dependent PEP carboxykinase

The Corynebacterium glutamicum (C. glutamicum) phosphoenolpyruvate carboxykinase (PCK) gene (pckA) was cloned into an Escherichia coli expression vector with a glutathione S-transferase (GST) tag. This recombinant DNA can produce highly overexpressed tagged protein in soluble form. This is the first report of the production of C. glutamicum PCK overexpressed in E. coli. The GST-fused PCK was purified using the glutathione-Sepharose 4B affinity column and the GST tag was removed in one-step. This one-step, easy purification method would be very useful for future mutational and structural studies. The molecular mass of the purified protein is approximately 68 kDa as confirmed by mass spectrometry and it is a monomeric enzyme. Also, the enzyme assays revealed that C. glutamicum PCK has a GTP-specific activity and that its activity is maximal in the presence of both Mn2+ and Mg2+.     

Effect of radish (Raphanus sativus Linn.) on thyroid status under conditions of varying iodine intake in rats

Cruciferous plants viz. cabbage, cauliflower, turnip, radish, mustard etc. that contain goitrogenic/antithyroid substances, constitute a portion of regular human diet. The effect of chronic feeding of fresh and cooked radish, R. sativus under varying state of iodine intake on morphological and functional status of thyroid in albino rats was evaluated by thyroid gland morphology and histology, thyroid peroxidase activity, serum triiodothyronine, thyroxine and thyrotropin levels. The consumption pattern of iodine and goitrogens of cyanogenic origin was evaluated by measuring urinary iodine and thiocyanate levels respectively. After chronic radish feeding, increased weight of thyroid gland, decreased thyroid peroxidase activity, reduced thyroid hormone profiles and elevated level of thyrotropin were observed resembling a relative state of hypoactive thyroid gland in comparison to control even after supplementation of adequate iodine.     

From natural products to polymeric derivatives of "eugenol": a new approach for preparation of dental composites and orthopedic bone cements

Polymers with eugenol moieties covalently bonded to the macromolecular chains were synthesized for potential application in orthopedic and dental cements. First, eugenol was functionalized with polymerizable groups. The synthetic methods employed afforded two different methacrylic derivatives, where the acrylic and eugenol moieties were either directly bonded, eugenyl methacrylate (EgMA), or separated through an oxyethylene group, ethoxyeugenyl methacrylate (EEgMA). A typical Fisher esterification reaction was used for the synthesis of EgMA and EEgMA, affording the desired monomers in 80% yields. Polymerization of each of the novel monomers, at low conversion, provided soluble polymers consisting of hydrocarbon macromolecules with pendant eugenol moieties. At high conversions only cross-linked polymers were obtained, attributed to participation of the allylic double bonds in the polymerization reaction. In addition, copolymers of each eugenol derivative with ethyl methacrylate (EMA) were prepared at low conversion, with the copolymerization reaction studied by assuming the terminal model and the reactivity ratios determined according to linear and nonlinear methods. The values obtained were r(EgMA) = 1.48, r(EMA) = 0.55 and r(EEgMA) = 1.22, r(EMA) = 0.42. High molecular weight polymers and copolymers were obtained at low conversion. Analysis of thermal properties revealed a T(g) of 95 degrees C for PEgMA and of 20 degrees C for PEEgMA and an increase in the thermal stability for the eugenol derivatives polymers and copolymers with respect to that of PEMA. Water sorption of the copolymers was found to decrease with the eugenol derivative content. Both monomers EgMA and EEgMA showed antibacterial activity against Streptococcus mutans, producing inhibition halos of 7 and 21 mm, respectively. Finally, cell culture studies revealed that the copolymers did not leach any toxic eluants and showed good cellular proliferation with respect to PEMA. This study thus indicates that the eugenyl methacrylate derivatives are potentially good candidates for dental and orthopedic cements.     

Generation of ceramide in murine macrophages infected with Leishmania donovani alters macrophage signaling events and aids intracellular parasitic survival

In the present study, we examined the involvement of intracellular ceramide in host pathogen interaction of BALB/c mouse peritoneal macrophages infected with the obligate intracellular protozoan, Leishmania donovani. Our findings indicate that the level of intracellular ceramide was enhanced as a result of the in vitro infection. While the elevated ceramide was largely due to de novo synthesis, activation of the sphingomyelinases was also observed. The enhanced ceramide was responsible for the downregulation of classical PKC activity, upregulation of calcium independent atypical PKC- expression and activity of calcium independent PKC. Ceramide also impaired the phosphorylation of MAPK. Evidently, ceramide suppressed the generation of nitric oxide during leishmanial infection and also facilitated the survival of leishmanial parasites in the intramacrophageal milieu. These data present newer insight to the signaling events in leishmania-infected murine macrophages, which might offer ceramide as a new therapeutic target in the future.     

In silico characterization of thermoactive, alkaline and detergent-stable lipase from a Staphylococcus aureus strain

Bacterial true lipases having thermo and alkaline stability are highly attractive for their industrial production of pharmaceuticals, agrochemicals, cosmetics, and flavour. Staphylococcus aureus lipase (SAL3) remains active at temperatures 40-60°C, with an optimum temperature of 55°C and an optimum pH of 9.5 stable over a range of 5-12. Detailed understanding of the structure and insight into the activity of such lipase would aid in engineering lipases that would function in the desired extreme industrial environments. In the present study, we carried out in silico characterization and structural modeling of SAL3 which is thermoactive, alkaline and detergent-stable. Comparison of SAL3 with other staphylococcal lipases indicates that SAL3 is a true lipase having the catalytic triad (residues Ser119, Asp310 & His352) and the calcium binding site (residues Asp351, Asp354, Asp359, Asp362 and Gly286). Conservation in sequence implies that interfacial activation mechanism is possible in SAL3 with the lid formed by helix (residues 180-196) and loop (residues 197-206). Three dimensional (3D) structure model of SAL3 has been predicted for the first time and aims at understanding its function and biochemical characteristics of possessing relatively high thermal and pH stability.     

Mycoplasma gallisepticum Lipid Associated Membrane Proteins Up-regulate Inflammatory Genes in Chicken Tracheal Epithelial Cells via TLR-2 Ligation through an NF-κB Dependent Pathway

Mycoplasma gallisepticum-mediated respiratory inflammation in chickens is associated with accumulation of leukocytes in the tracheal submucosa. However the molecular mechanisms underpinning these changes have not been well described. We hypothesized that the initial inflammatory events are initiated upon ligation of mycoplasma lipid associated membrane proteins (LAMP) to TLRs expressed on chicken tracheal epithelial cells (TEC). To test this hypothesis, live bacteria or LAMPs isolated from a virulent (R_low) or a non-virulent (R_high) strain were incubated with primary TECs or chicken tracheae ex vivo. Microarray analysis identified up-regulation of several inflammatory and chemokine genes in TECs as early as 1.5 hours post-exposure. Kinetic analysis using RT-qPCR identified the peak of expression for most genes to be at either 1.5 or 6 hours. Ex-vivo exposure also showed up-regulation of inflammatory genes in epithelial cells by 1.5 hours. Among the commonly up-regulated genes were IL-1β, IL-6, IL-8, IL-12p40, CCL-20, and NOS-2, all of which are important immune-modulators and/or chemo-attractants of leukocytes. While these inflammatory genes were up-regulated in all four treatment groups, R_low exposed epithelial cells both in vitro and ex vivo showed the most dramatic up-regulation, inducing over 100 unique genes by 5-fold or more in TECs. Upon addition of a TLR-2 inhibitor, LAMP-mediated gene expression of IL-1β and CCL-20 was reduced by almost 5-fold while expression of IL-12p40, IL-6, IL-8 and NOS-2 mRNA was reduced by about 2–3 fold. Conversely, an NF-κB inhibitor abrogated the response entirely for all six genes. miRNA-146a, a negative regulator of TLR-2 signaling, was up-regulated in TECs in response to either R_low or R_high exposure. Taken together we conclude that LAMPs isolated from both R_high and R_low induced rapid, TLR-2 dependent but transient up-regulation of inflammatory genes in primary TECs through an NF-κB dependent pathway.