Culture medium optimization for improved puerarin production by cell suspension cultures of Pueraria tuberosa (Roxb. ex Willd.) DC.

The effects of carbon, nitrogen, phosphate, and copper on cell growth and production of the isoflavone puerarin by suspension cultures of Pueraria tuberosa (Roxb. ex. Willd.) DC were investigated. Among the various sugars evaluated (glucose, galactose, fructose, maltose, and sucrose), use of sucrose in the medium led to the maximum accumulation of puerarin. A sucrose-feeding strategy in which additional sucrose was added to the flasks 15?d into the culture cycle stimulated both cell biomass and puerarin production. The maximum production of puerarin was obtained when a concentration balance of 20:60?mM NH ₄ ⁺ /NO ₃ ^? was used as the nitrogen source. Alteration in the concentration balance of nitrogen components (NH ₄ ⁺ /NO ₃ ^? 60:20?mM) or the use of either NH ₄ ⁺ or NO ₃ ^? alone decreased biomass production and puerarin accumulation compared with the control culture (NH ₄ ⁺ /NO ₃ ^? 20:20?mM). High amounts of phosphate (2.5 and 5?mM) in the medium inhibited puerarin production whereas 0.625?mM phosphate promoted puerarin production (68.3???g/g DW on day?25). An increase in Cu²⁺ concentration from 0.025 to 0.05?mg/l in the P. tuberosa cell culture medium resulted in a 2.2-fold increase in puerarin production (up to 141???g/g DW on day?25) but reduced cell culture biomass.     

A 12-deoxywithastramonolide-rich somaclonal variant in Withania somnifera (L.) Dunal—molecular cytogenetic analysis and significance as a chemotypic resource

Withania somnifera, commonly known as ashwagandha or Indian ginseng, is a valuable medicinal plant, synthesizing a wide array of pharmacologically active secondary metabolites known as withanolides. In this study, we investigated variation among 54 regenerated plants attained through indirect organogenesis from leaf explants. Organogenic calli were induced on Murashige and Skoog medium containing 2?mg?l^?1 kinetin and 1?mg?l^?1 indole-3-butyric acid. High-performance liquid chromatography was used for quantitative determination of the major withanolides in the somaclones. One somaclone (WS-R-1) showed significantly higher accumulation of 12-deoxywithastramonolide (WS-12D; 0.516%) compared to the explant donor mother plant (0.002%). The incidence of somaclonal variation at the cytological level was investigated by studying mitosis and meiosis in relation to chromosome number and structural organization. There were no alterations in chromosome phenotypes, somatic chromosome count, or meiotic behavior. Fidelity at genomic level was evaluated by random amplification of polymorphic DNA (RAPD) analyses, which revealed multiple genetic polymorphisms between the WS-12D over-producing somaclone and the explant donor mother plant. This study demonstrates the capability of inducing chemotypic variability for the development of high-yielding clones due to molecular instability in W. somnifera using an in vitro approach.     

Identification, purification and characterization of laterosporulin, a novel bacteriocin produced by Brevibacillus sp. strain GI-9

Background

Bacteriocins are antimicrobial peptides that are produced by bacteria as a defense mechanism in complex environments. Identification and characterization of novel bacteriocins in novel strains of bacteria is one of the important fields in bacteriology.

Methodology/Findings

The strain GI-9 was identified as Brevibacillus sp. by 16 S rRNA gene sequence analysis. The bacteriocin produced by strain GI-9, namely, laterosporulin was purified from supernatant of the culture grown under optimal conditions using hydrophobic interaction chromatography and reverse-phase HPLC. The bacteriocin was active against a wide range of Gram-positive and Gram-negative bacteria. MALDI-TOF experiments determined the precise molecular mass of the peptide to be of 5.6 kDa and N-terminal sequencing of the thermo-stable peptide revealed low similarity with existing antimicrobial peptides. The putative open reading frame (ORF) encoding laterosporulin and its surrounding genomic region was fished out from the draft genome sequence of GI-9. Sequence analysis of the putative bacteriocin gene did not show significant similarity to any reported bacteriocin producing genes in database.

Conclusions

We have identified a bacteriocin producing strain GI-9, belonging to the genus Brevibacillus sp. Biochemical and genomic characterization of laterosporulin suggests it as a novel bacteriocin with broad spectrum antibacterial activity.     

Cannabinoid Receptor 1 Signaling in Embryo Neurodevelopment

In utero exposure to tetrahydrocannabinol, the psychoactive component of marijuana, is associated with an increased risk for neurodevelopmental defects in the offspring by interfering with the functioning of the endocannabinoid (eCB) system. At the present time, it is not clearly known whether the eCB system is present before neurogenesis. Using an array of biochemical techniques, we analyzed the levels of CB1 receptors, eCBs (AEA and 2‐AG), and the enzymes (NAPE‐PLD, DAGLα, DAGLβ, MAGL, and FAAH) involved in the metabolism of the eCBs in chick and mouse models during development. The findings demonstrate the presence of eCB system in early embryo before neurogenesis. The eCB system might play a critical role in early embryogenesis and there might be adverse developmental consequences of in utero exposure to marijuana and other drugs of abuse during this period.     

NeMedPlant: a database of therapeutic applications and chemical constituents of medicinal plants from north-east region of India

The North-East region of India is one of the twelve mega biodiversity region, containing many rare and endangered species. A curated database of medicinal and aromatic plants from the regions called NeMedPlant is developed. The database contains traditional, scientific and medicinal information about plants and their active constituents, obtained from scholarly literature and local sources. The database is cross-linked with major biochemical databases and analytical tools. The integrated database provides resource for investigations into hitherto unexplored medicinal plants and serves to speed up the discovery of natural productsbased drugs. AVAILABILITY: The database is available for free at http://bif.uohyd.ac.in/nemedplant/orhttp://202.41.85.11/nemedplant/     

System modeling and identification in indicator dilution method for assessment of Ejection Fraction and Pulmonary Blood Volume

Clinically relevant cardiovascular parameters, such as pulmonary blood volume (PBV) and ejection fraction (EF), can be assessed through indicator dilution techniques. Among these techniques, which are typically invasive due to the need for central catheterization, contrast ultrasonography provides a new emerging minimally invasive option. PBV and EF are then measured by a dilution system identification algorithm after detection of multiple dilution curves by an ultrasound scanner. In this paper, dilution systems are represented by parametric models. Since the measured indicator dilution curves (IDCs) are corrupted by measurement artifacts and outliers, the use of conventional least square error (LSE) estimator for estimating system parameters is not optimal. Different estimators are therefore proposed for estimating the system parameters. Comparison of these estimators with the LSE estimator in assessing EF and PBV is performed on simulated, in vitro and patient data. The results show that the proposed total least absolute deviation estimator (TLAD) outperforms other estimators. The measured IDCs are highly corrupted by noise, which affect the estimation of EF and PBV. Therefore, a two stage denoising method capable of removing outliers is also proposed for removing noise in IDCs.     

Post-Operative Hypertension after Total Knee Arthroplasty and the Effects on Transfusion Rates

Transfusions are a cause of significant patient morbidity as well as expense. Anesthesia literature has examined controlled intraoperative hypotension as a means for reducing blood loss and transfusions. Our hypothesis is that inversely increased blood pressure post-operatively would then lead to increased blood loss and transfusions.We examined 105 consecutive patients who underwent TKA. We found a significant odds ratio of 1.123 for pre-operative hematocrit. For post-operative blood pressure, we calculated an insignificant odds ratio of 1.007, proving no relationship between post-operative blood pressure and transfusions.This is the first study to examine increased post-operative blood pressure''s contribution to transfusion rates. Although we confirmed that low pre-operative hematocrit contributes to increased transfusions, we did not find a relationship between post-operative blood pressure and transfusions.     

Effect of different levels of selenium supplementation on growth rate, nutrient utilization, blood metabolic profile, and immune response in lambs

Eighteen male lambs (8-9 months of age, 25.00 +/- 0.90 kg body weight) were divided into three groups of six animals in each and fed a total mixed ration (TMR) containing concentrate mixture (30% maize grain, 27% soybean meal, 40% wheat bran, 2% mineral mixture, and 1% common salt) and wheat straw in 65:35 ratio and supplemented with selenium (Se) as sodium selenite at 0 (T1, control), 0.15 (T2), and 0.30 ppm (T3) levels. Experimental feeding was done for a period of 90 days including a 6-day metabolism trial. To assess the growth performance, lambs were weighed every 15 days throughout the experimental period. All the lambs were intramuscularly inoculated with a single dose (2 ml) of haemorrhagic septicaemia oil adjuvant vaccine on 0 day to evaluate the humoral immune response. Blood samples were collected on 0 day and thereafter at 30 days interval. Results revealed that supplementation of Se both at 0.15 and 0.30 ppm levels had no significant (P > 0.05) effect on intake and digestibility of dry matter, organic matter, crude protein (CP), ether extract, neutral detergent fiber, acid detergent fiber, and hemicellulose; balances of calcium and phosphorus; and level and intake of digestible CP and total digestible nutrients. Se supplementation also had no significant (P > 0.05) effect on the levels of serum total cholesterol, total protein, albumin, globulin, albumin/globulin ratio, tri-iodothyronine (T(3)), thyroxine (T(4)), and T(4)/T(3) ratio; and serum glutamate oxaloacetate transaminase and serum glutamate pyruvate transaminase enzyme activity in the lambs. However, there was a significant (P < 0.05) increase in the plasma Se levels, red blood cell glutathione peroxidase enzyme activity, and humoral immune response in both the Se-supplemented groups. Feed (TMR) required per kilogram gain was less by 11.1% and 16.5% in groups T2 and T3, respectively, as compared to control (T1) group. Average daily gain was highest (108.5 g) in group T3, followed by group T2 (98.2 g), and lowest (89.06 g) in the control group (T1). These results indicated that supplementation of 0.15 and 0.3 ppm Se in the diet (having 0.19 ppm Se) of lambs significantly improves their immune response and antioxidant status.