Taxol Determination from Pestalotiopsis pauciseta, a Fungal Endophyte of a Medicinal Plant

Taxol is the most effective antitumor agent developed in the past three decades. It has been used for effective treatment of a variety of cancers. A taxol-producing endophytic fungus Pestalotiopsis pauciseta (strain CHP-11) was isolated from the leaves of Cardiospermum helicacabum and screened for taxol production. The fungus was identified based on the morphology of the fungal culture and the characteristics of the spores and screened for taxol production. The amount of taxol produced by this endophytic fungus was quantified by HPLC and it produced 113.3 mg/L, thus the fungus can serve as a potential material for fungus engineering to improve taxol production. This fungal taxol also had strong anticancer activity against some cancer cells viz., BT 220, H116, Int 407, HL 251 and HLK 210 tested by Apoptotic assay and it is indicated that with the increase of taxol concentration from 0.005–0.05 mmol/L, taxol induced increased cell death through apoptosis.     

First record of the family Thamnocephalidae (Crustacea: Anostraca) from Southeast Asia and description of a new species of Branchinella

During a two-year investigation of freshwater microcrustaceans in Thailand, an undescribed species of fairy shrimp of the genus Branchinella Sayce, 1903 in the family Thamnocephalidae, was discovered. This is the first record of the genus and the family from Southeast Asia and the second anostracan reported from Thailand. The new species, Branchinella thailandensis n. sp. appears to be morphologically intermediate between B. kugenumaensis (Ishikawa, 1895) from Japan and east China, and B. madurai Raj, 1951 from India and Pakistan. B. thailandensis n. sp. was collected from several temporary ponds in 11 provinces of the northeast and the central Thailand, often co-occurring with Streptocephalus sirindhornae Sanoamuang, Murugan, Weekers & Dumont, 2000.     

Evidence for a novel phosphopantetheinyl transferase domain in the polyketide synthase for enediyne biosynthesis

The polyketide synthase associated with the biosynthesis of enediyne-containing calicheamicin contains a putative phosphopantetheinyl transferase (PPTase) domain. By cloning and expressing the C-terminal region of the polyketide synthase and in vitro phosphopantetheinylation assay, we found that the PPTase domain exhibits preferred substrate specificity towards acyl and peptidyl carrier proteins in fatty acid and non-ribosomal peptide synthesis over its cognate partner. We also found evidence suggesting that the PPTase domain adopts a pseudo-trimeric structure, distinct from the pseudo-dimeric structure of type II PPTases. The results revealed a novel type of PPTase with unique structure and substrate specificity, and suggested that the polyketide synthase probably acquired the PPTase domain from a primary metabolic pathway in evolution.     

Effect of tetrahydrocurcumin on insulin receptor status in type 2 diabetic rats: studies on insulin binding to erythrocytes

Curcumin is the most active component of turmeric. It is believed that curcumin is a potent antioxidant and anti-inflammatory agent. Tetrahydrocurcumin (THC) is one of the major metabolites of curcumin, and exhibits many of the same physiological and pharmacological activities as curcumin and, in some systems, may exert greater antioxidant activity than curcumin. Using circulating erythrocytes as the cellular mode, the insulin-binding effect of THC and curcumin was investigated. Streptozotocin (STZ)-nicotinamide-induced male Wistar rats were used as the experimental models. THC (80 mg/kg body weight) was administered orally for 45 days. The effect of THC on blood glucose, plasma insulin and insulin binding to its receptor on the cell membrane of erythrocytes were studied. Mean specific binding of insulin was significantly lowered in diabetic rats with a decrease in plasma insulin. This was due to a significant decrease in mean insulin receptors. Erythrocytes from diabetic rats showed a decreased ability for insulin-receptor binding when compared with THC-treated diabetic rats. Scatchard analysis demonstrated that the decrease in insulin binding was accounted for by a decrease in insulin receptor sites per cell, with erythrocytes of diabetic rats having less insulin receptor sites per cell than THC-treated rats. High affinity (K d1), low affinity (K d2) and kinetic analyses revealed an increase in the average receptor affinity of erythrocytes from THC-treated rats compared with those of diabetic rats. These results suggest that acute alteration of the insulin receptor on the membranes of erythrocytes occurred in diabetic rats. Treatment with THC significantly improved specific insulin binding to the receptors, with receptor numbers and affinity binding reaching near-normal levels. Our study suggests the mechanism by which THC increases the number of total cellular insulin binding sites resulting in a significant increase in plasma insulin. The effect of THC is more prominent than that of curcumin.     

Bioreduction of hexavalent chromium by <Emphasis Type="Italic">Pseudomonas stutzeri</Emphasis> L1 and <Emphasis Type="Italic">Acinetobacter baumannii</Emphasis> L2

Bioreduction of the very toxic hexavalent chromium ion [Cr(VI)] to the non-toxic trivalent chromium ion [Cr(III)] is a key remediation process in chromium-contaminated sites. In this study, we investigated the bioreduction of Cr(VI) by Pseudomonas stutzeri L1 and Acinetobacter baumannii L2. The optimum pH (5–10), temperature (27, 37 and 60 °C) and initial chromium Cr(VI) concentration (100–1000 mg L^?1) for Cr(VI) reduction by strains L1 and L2 were determined using the diphenylcarbazide method. In the presence of L1 and L2, the bioreduction rate of Cr(VI) was 40–97 and 84–99%, respectively. The bioreduction of Cr(VI) by L2 was higher, reaching up to 84%—than that by L1. The results showed that strain L2 was able to survive even if exposed to 1000 mg L^?1 of Cr(VI) and that this tolerance to the effects of Cr(VI) was linked to the activity of soluble enzyme fractions. Overall, A. baumannii L2 would appear to be a potent Cr(VI)-tolerant candidate for the bioremediation of chromium (VI)-contaminated wastewater effluent.     

A simple formalism on dynamics of proteins on potential energy landscapes

We present a simple formalism for the dynamics of proteins on a potential energy landscape, using connectedness of configurational domains as an order parameter. This formalism clearly shows that the energy bias required to form a unit correct contact toward the native configuration of a two-state folder, to overcome Levinthal's paradox, is E(bias) congruent with RT ln 2. This result agrees well with earlier studies and indicates that the bias is mainly due to hydrophobic interaction. Further investigations have shown that the landscape funnel could be experimentally mapped onto a two-dimensional space formed by denaturant concentration and the connectedness of configurational domains. The theoretical value of the depth-of-folding funnel in terms of denaturant concentration has been calculated for a model protein (P450cam), which agrees well with the experimental value. Using our model, it is also possible to explain the turnover nature of heat-capacity change upon unfolding of proteins and the existence of enthalpy and entropy convergence temperatures during unfolding without any strict assumptions as proposed in earlier studies.     

Design of an Inflammation-Sensitive Polyelectrolyte-Based Topical Drug Delivery System for Arthritis

The most successful treatment strategy for arthritis is intra-articular injections that are costly and have reduced patient compliance. The purpose of the current study was to develop an inflammation-sensitive system for topical drug administration. Multi-macromolecular alginate-hyaluronic acid-chitosan (A-H-C) polyelectrolyte complex nanoparticles, loaded with indomethacin were developed employing pre-gel and post-gel techniques in the presence of dodecyl-l-pyroglutamate (DLP). In addition to in vitro studies, in silico simulations were performed to affirm and associate the molecular interactions inherent to the formulation of core all-natural multi-component biopolymeric architectures composed of an anionic (alginate), a cationic (chitosan), and an amphi-ionic polyelectrolytic (hyaluronic acid) macromolecule. The results demonstrated that DLP significantly influenced the size of the synthesized nanoparticles. Drug-content analysis revealed higher encapsulation efficiency (77.3%) in the presence of DLP, irrespective of the techniques used. Moreover, in vitro drug release studies showed that indomethacin release from the nanosystem was significantly improved (98%) in Fenton’s reagent. Drug permeation across a cellulose membrane using a Franz diffusion cell system showed an initial surge flux (0.125 mg/cm^?2/h), followed by sustained release of indomethacin for the post-gel nanoparticles revealing its effective skin permeation efficiency. In conclusion, the study presents novel nanoparticles which could effectively encapsulate and deliver hydrophobic drugs to the target site, particularly for arthritis.     

Urinary catheter indwelling clinical pathogen biofilm formation,exopolysaccharide characterization and their growth influencing parameters

Self-reproducing microbial biofilm community mainly involved in the contamination of indwelling medical devices including catheters play a vital role in nosocomial infections. The catheter-associated urinary tract infection (CA-UTI) causative Staphylococcus aureus, Enterobacter faecalis, and Pseudomonas aeruginosa were selectively isolated, their phenotypic as well as genotypic biofilm formation, production and monomeric sugar composition of EPS as well as sugar, salt, pH and temperature influence on their in vitro biofilm formation were determined. From 50 culture positive urinary catheters S. aureus (24%), P. aeruginosa (18%), E. faecalis (14%) and others (44%) were isolated. The performed assays revealed their varying biofilm forming ability. The isolated S. aureus ica, E. faecalis esp, and P. aeruginosa cup A gene sequencing and phylogenetic analysis showed their close branching and genetic relationship. The analyzed sugar, salt, pH, and temperature showed that the degree of CA-UTI isolates biofilm formation is an environmentally sensitive process. EPS monosaccharide HPLC analysis showed the presence of neutral sugars (ng/μl) as follows: glucose (P. aeruginosa: 44.275; E. faecalis: 4.23), lactose (P. aeruginosa: 7.29), mannitol (P. aeruginosa: 2.53; S. aureus: 2.62; E. faecalis: 2.054) and maltose (E. faecalis: 7.0042) revealing species-specific presence and variation. This study may have potential clinical relevance for the easy diagnosis and management of CA-UTI.     

Phylogenetic Diversity and In Vitro Susceptibility Profiles of Human Pathogenic Members of the <Emphasis Type="Italic">Fusarium fujikuroi</Emphasis> Species Complex Isolated from South India

Availability of molecular methods, gene sequencing, and phylogenetic species recognition have led to rare fungi being recognized as opportunistic pathogens. Fungal keratitis and onychomycosis are fairly common mycoses in the tropics, especially among outdoor workers and enthusiasts. The frequently isolated etiological agents belong to genera Candida, Aspergillus, and Fusarium. Within the genus Fusarium, known to be recalcitrant to prolonged antifungal treatment and associated with poor outcome, members of the Fusarium solani species complex are reported to be most common, followed by members of the Fusarium oxysporum SC and the Fusarium fujikuroi SC (FFSC). Morphological differentiation among the various members is ineffective most times. In the present study, we describe different species of the FFSC isolated from clinical specimen in south India. All twelve isolates were characterized up to species level by nucleic acid sequencing and phylogenetic analysis. The molecular targets chosen were partial regions of the internal transcribed spacer rDNA region, the panfungal marker and translation elongation factor-1α gene, the marker of choice for Fusarium speciation. Phylogenetic analysis was executed using the Molecular Evolutionary Genetics Analysis software (MEGA7). In vitro susceptibility testing against amphotericin B, voriconazole, posaconazole, natamycin, and caspofungin diacetate was performed following the CLSI M38-A2 guidelines for broth microdilution method. The twelve isolates of the FFSC were F. verticillioides (n = 4), F. sacchari (n = 3), F. proliferatum (n = 2), F. thapsinum (n = 1), F. andiyazi (n = 1), and F. pseudocircinatum (n = 1). To the best of our knowledge, this is the first report of F. andiyazi from India and of F. pseudocircinatum as a human pathogen worldwide. Natamycin and voriconazole were found to be most active agents followed by amphotericin B. Elderly outdoor workers figured more among the patients and must be recommended protective eye wear.