Fungal biodegradation of phthalate plasticizer in situ

This unique study describes how Aspergillus japonicus, Penicillium brocae and Purpureocillium lilacinum, three novel isolates of our laboratory from heavily plastics-contaminated soil completely utilized the plasticizer di(2-ethylhexyl)phthalate (DEHP) bound to PVC blood storage bags (BB) in simple basal salt medium (BSM) by static submerged growth (28 °C). Initial quantification as well as percentage utilization of DEHP blended to BB were estimated periodically by extracting it into n-hexane. A two-stage cultivation strategy was employed for the complete mycoremediation of DEHP from BB in situ. During the first growth stage, about two-third parts of total (33.5 % w/w) DEHP bound to BB were utilized in two weeks, accompanied by increased fungal biomass (~0.15–0.32 g per g BB) and sharp declining (to ~3) of initial pH (7.2). At this stagnant growth state (low pH), spent medium was replaced by fresh BSM (pH, 7.2), and thus in the second stage the remaining DEHP (one-third) in BB was utilized completely. The ditches and furrows seen from the topology of the BB as seen by the 3D AFM image further confirmed the bioremediation of DEHP physically bound to BB in situ. Of the three mycelial fungi employed, P. lilacinum independently showed highest efficiency for the complete utilization of DEHP bound to BB, whose activity was comparable to that of the consortium comprising all the three fungi described herein. To sum up, the two-stage cultivation strategy demonstrated in this study shows that a batch process would efficiently remediate the phthalic acid esters blended in plastics on a large scale, and thus it offers potentials for the management of plastics wastes.     

Species richness in agamid lizards: chance,body size,sexual selection or ecology?

Why does species richness vary so greatly across lineages? Traditionally, variation in species richness has been attributed to deterministic processes, although it is equally plausible that it may result from purely stochastic processes. We show that, based on the best available phylogenetic hypothesis, the pattern of cladogenesis among agamid lizards is not consistent with a random model, with some lineages having more species, and others fewer species, than expected by chance. We then use phylogenetic comparative methods to test six types of deterministic explanation for variation in species richness: body size, life history, sexual selection, ecological generalism, range size and latitude. Of eight variables we tested, only sexual size dimorphism and sexual dichromatism predicted species richness. Increases in species richness are associated with increases in sexual dichromatism but reductions in sexual size dimorphism. Consistent with recent comparative studies, we find no evidence that species richness is associated with small body size or high fecundity. Equally, we find no evidence that species richness covaries with ecological generalism, latitude or range size.     

Molecular phylogeny of asexual entomopathogenic fungi with special reference to Beauveria bassiana and Nomuraea rileyi

The phylogenetic lineage, taxonomic affiliation and interrelationships of important asexual entomopathogenic fungal genera were studied using the sequences of partial regions of β-tubulin and rRNA genes. The species structures of Beauveria bassiana and Nomuraea rileyi were also investigated. A total of 147 fungal entries covering 94 species were analysed. Phylogenetic analysis placed all the asexual entomopathogenic fungal species analysed, in the family Clavicipitaceae of the order Hypocreales of Ascomycota. Deep phylogenetic lineages were observed in B. bassiana iterating the complex nature of this species. Some of the isolates assigned to this species separated out more distinctly than morphologically distinguishable genera. Cryptic speciation was also evident in N. rileyi. It is concluded that the asexual fungi with entomopathogenic habit arose from a single lineage in sexual Clavicipitaceae.     

Water soluble vitamin E (TMG) as a radioprotector

Tocopherol monoglucoside (TMG), a water soluble derivative of vitamin E offers protection against deleterious effects of ionizing radiation, both under in vivo and in vitro conditions, to biological systems. TMG was found to be a potent antioxidant and an effective free radical scavenger. It forms a phenoxyl radical similar to trolox upon reaction with various one-electron oxidants. TMG protected DNA from radiation-induced strand breaks. It also protected thymine glycol formation induced by gamma-radiation. Gamma-radiation-induced loss of viability of EL-tumor cells and peroxidation of lipids in microsomal and mitochondrial membranes were prevented by TMG. TMG was nontoxic to mice when administered orally up to 7.0 g/kg body weight. The LD50 dose of TMG for ip administration in mice was 1.15 g/kg body wt. In rats, following oral and ip administration of TMG, the absorption (distribution) half lives were 5.8 and 3.0 min respectively and elimination half lives were 6.7 and 3.1 min respectively. Embryonic mortality resulting from exposure of pregnant mice to ionizing radiation (2 Gy) was reduced by 75% by ip administration of TMG (0.6 g/kg, body wt) prior to irradiation. TMG offered protection to mice against whole body gamma-radiation-induced lethality and weight loss. The LD50(30) of mice increased from 6 to 6.72 Gy upon post irradiation administration of a single dose of TMG (0.6 g/kg, body wt) by ip.     

Monitoring the Disulfide Bond Formation of a Cysteine-Rich Repeat Protein from Helicobacter pylori in the Periplasm of Escherichia coli

Helicobacter pylori infection increases the risk of cardiovascular diseases besides leading to duodenal and gastric peptic ulcerations. H. pylori cysteine-rich protein B (HcpB) is a disulfide-rich repeat protein that belongs to the family of Sel1-like repeat proteins. HcpB contains four pairs of anti-parallel alpha helices that fold into four repeats with disulfide bonds bridging the helices of each repeat. Recent in vitro oxidative refolding of HcpB identified that the formation and folding of the disulfide bond in the N-terminal repeat are the rate limiting step. Here we attempted to understand the disulfide formation of HcpB in the periplasm of Escherichia coli. The protein was expressed in wild type (possessed enzymes DsbA, B, C, and D) and knock out (Dsb enzymes deleted one at a time) E. coli strains. The soluble part of the periplasm when analyzed by SDS-PAGE and Western Blot showed that the wild type and DsbC/D knock out strains contained native oxidized HcpB while the protein was absent in the DsbA/B knock out strains. Hence the recombinant expression of HcpB in E. coli requires DsbA and DsbB for disulfide bond formation and it is independent of DsbC and DsbD. Prolonged cell growth resulted in the proteolytic degradation of the N-terminal repeat of HcpB. The delayed folding of the N-terminal repeat observed during in vitro oxidative refolding could be the reason for the enhanced susceptibility to proteolytic cleavage in the periplasm. In summary, a good correlation between in vivo and in vitro disulfide bond formation of HcpB is observed.     

Does dietary magnesium modulate blood lipids?

In a randomized, single-blind, controlled study (400 patients aged 25-63 yr; 374 males, 26 females), 206 subjects were administered a magnesium-rich diet, and 194 subjects their usual diet, for 6 wk. Age, sex, body weight, hypertension, hyperlipidemia, smoking, obesity, diuretic therapy, and diabetes were comparable between the two groups, as were laboratory data at entry to the study. Intervention-group A received a significantly higher amount of dietary magnesium and potassium compared to group B, which received its usual diet. After 6 wk, there was a significant fall in total serum cholesterol (228.5 +/- 46.2 mg/dL), LDL cholesterol 146.5 +/- 75.5 mg/dL), and triglyceride (143.8 +/- 40.5 mg/dL) in group A compared to serum cholesterol (242.5 +/- 58.2 mg/dL), LDL cholesterol (157.0 +/- 78.4 mg/dL), and triglyceride (156.5 +/- 60.0 mg/dL) at entry to study, but no such changes in group-B subjects. HDL cholesterol showed a marginal mean decrease of 0.8 mg/dL in group B and a 2.5 mg/dL increase in group A. The changes in blood lipids were consistent with an increased intake of magnesium and with a rise in serum levels. Although a general blood-lipid-reducing effect of such a diet cannot be excluded, it is possible that dietary magnesium may have contributed to the reduction of total serum cholesterol, LDL cholesterol, and triglyceride, and the marginal rise in HDL cholesterol. More studies with longer follow-up periods are needed to confirm this observation.     

Differential effects of cholesterol and 7-dehydrocholesterol on ligand binding of solubilized hippocampal serotonin1A receptors: implications in SLOS

The serotonin1A receptor is an important member of the G-protein coupled receptor family, and is involved in the generation and modulation of a variety of cognitive, behavioral, and developmental functions. Solubilization of the hippocampal serotonin1A receptor by 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) is accompanied by loss of membrane cholesterol which results in a reduction in specific agonist binding activity. Replenishment of cholesterol to solubilized membranes restores the cholesterol content of the membrane and significantly enhances specific agonist binding activity. In order to test the stringency of the requirement of cholesterol in this process, we solubilized native hippocampal membranes followed by replenishment with 7-dehydrocholesterol (7-DHC). 7-DHC is an immediate biosynthetic precursor of cholesterol differing only in a double bond at the 7th position in its sterol ring. Our results show, for the first time, that replenishment of solubilized hippocampal membranes with 7-DHC does not restore ligand binding activity of the serotonin1A receptor, in spite of recovery of the overall membrane order. This observation shows that the requirement for restoration of ligand binding activity is more stringent than the requirement for the recovery of overall membrane order. These novel results have potential implications in understanding the interaction of membrane sterols with this important neuronal receptor under pathogenic conditions such as the Smith-Lemli-Opitz syndrome.     

The endosomal adaptor protein APPL1 impairs the turnover of leading edge adhesions to regulate cell migration

Cell migration is a complex process that requires the integration of signaling events that occur in distinct locations within the cell. Adaptor proteins, which can localize to different subcellular compartments, where they bring together key signaling proteins, are emerging as attractive candidates for controlling spatially coordinated processes. However, their function in regulating cell migration is not well understood. In this study, we demonstrate a novel role for the adaptor protein containing a pleckstrin-homology (PH) domain, phosphotyrosine-binding (PTB) domain, and leucine zipper motif 1 (APPL1) in regulating cell migration. APPL1 impairs migration by hindering the turnover of adhesions at the leading edge of cells. The mechanism by which APPL1 regulates migration and adhesion dynamics is by inhibiting the activity of the serine/threonine kinase Akt at the cell edge and within adhesions. In addition, APPL1 significantly decreases the tyrosine phosphorylation of Akt by the nonreceptor tyrosine kinase Src, which is critical for Akt-mediated cell migration. Thus, our results demonstrate an important new function for APPL1 in regulating cell migration and adhesion turnover through a mechanism that depends on Src and Akt. Moreover, our data further underscore the importance of adaptor proteins in modulating the flow of information through signaling pathways.     

Ecological variations in adult life table attributes of Aedes aegypti (L.) from the desert and coastal regions of India

The ecological variation in biological and adult life-table attributes of two populations of Aedes aegypti (Diptera: Culicidae) from the desert (Jodhpur) and coastal (Kolkata) regions of India are assessed to understand the reproductive and survival strategies. The results showed that females lived longer than males in both strains. The desert strain was more r-strategist because of its higher intrinsic rate of increase (r_m = 0.23), finite rate of increase (λ = 1.25), lower life expectancy of males (7.9 days) and females (14.4 days), mean generation time (T = 19.2 days) and doubling time (DT = 3.0 days). However, there was no difference in net reproductive rate (R₀) between the desert and coastal strains. The coastal strain showed a longer female life expectancy (22.0 days) than the desert strain (14.4 days). However, the fecundity (eggs/female/day) was lower in the coastal strain (11.4) than in the desert strain (15.1). Conclusively, the desert (Jodhpur) strain is adapted to a better r-strategy than the coastal (Kolkata) strain of Ae. aegypti, which might be helpful to flourish in harsh environmental conditions. This study may provide accurate predictions of Ae. aegypti population dynamics for vector management.