Reactive centre loop mutants of α-1-antitrypsin reveal position-specific effects on intermediate formation along the polymerization pathway

The common severe Z mutation (E342K) of α₁-antitrypsin forms intracellular polymers that are associated with liver cirrhosis. The native fold of this protein is well-established and models have been proposed from crystallographic and biophysical data for the stable inter-molecular configuration that terminates the polymerization pathway. Despite these molecular ‘snapshots’, the details of the transition between monomer and polymer remain only partially understood. We surveyed the RCL (reactive centre loop) of α₁-antitrypsin to identify sites important for progression, through intermediate states, to polymer. Mutations at P₁₄P₁₂ and P₄, but not P₁₀P₈ or P₂P_1′, resulted in a decrease in detectable polymer in a cell model that recapitulates the intracellular polymerization of the Z variant, consistent with polymerization from a near-native conformation. We have developed a FRET (Förster resonance energy transfer)-based assay to monitor polymerization in small sample volumes. An in vitro assessment revealed the position-specific effects on the unimolecular and multimolecular phases of polymerization: the P₁₄P₁₂ region self-inserts early during activation, while the interaction between P₆P₄ and β-sheet A presents a kinetic barrier late in the polymerization pathway. Correspondingly, mutations at P₆P₄, but not P₁₄P₁₂, yield an increase in the overall apparent activation energy of association from ~360 to 550 kJ mol⁻¹.     

Chromium-tolerant bacteria isolated from industrial effluents and their use in detoxication of hexavalent chromium

Three bacterial strains were isolated from effluents of leather (CMBL Cr13, CMBL Cr14) and steel (CMBL Cr15) industries for their possible use in chromium(VI) detoxication of industrial waste. CMBL Cr13 was found to tolerate chromium(VI) up to a concentration of 45 g/L in the medium, while CMBL Cr14 and CMBL Cr15 could tolerate up to 40 g/L. These bacteria were also checked for resistance to other metals. They resisted a lead concentration of 1 g/L and cadmium concentration of 550 mg/L in the medium. They showed optimum growth at pH 7.3–7.5 at a temperature of 35–37°C. Cr^VI-reducing ability of the three strains ranged from 70 to 80% after 3 d of incubation. The possible use of these bacteria in environmental cleanup is discussed.     

Methoprene modulates the effect of diet on male melon fly,Bactrocera cucurbitae,performance at mating aggregations

The effect of access to dietary protein (P) (hydrolyzed yeast) and/or treatment with a juvenile hormone analogue, methoprene (M), (in addition to sugar and water) on male aggregation (lekking) behaviour and mating success was studied in a laboratory strain of the melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae). Six‐day‐old males were treated with (1) protein and methoprene (M+P+), (2) only protein (M?P+), or (3) only methoprene (M+P?), and compared with 14‐day‐old sexually mature untreated males (M?P?). The lekking behaviour of the four groups of males when competing for virgin sexually mature females (14 –16 days old) was observed in field cages. The following parameters were measured at male aggregations: lek initiation, lek participation, males calling, male–male interaction, female acceptance index, and mating success. For all these parameters, the M+P+ males significantly outperformed the other males. Moreover, for all parameters, there was a similar trend with M+P+ > M?P+ > M?P? > M+P?. More M+P+ males called and initiated and participated in lek activities than all other types of male, which resulted in higher mating success. They had also fewer unsuccessful copulation attempts than their counterparts. Whereas treatment with methoprene alone had a negative effect in young males with only access to sugar, access to dietary protein alone significantly improved young male sexual performance; moreover, the provision of methoprene together with protein had a synergistic effect, improving further male performance at leks. The results are of great relevance for enhancing the application of the sterile insect technique (SIT) against this pest species. The fact that access to dietary protein and treatment of sterile males with methoprene improves mating success means that SIT cost‐effectiveness is increased, as more released males survive to sexual maturity.     

Prospect of phytoaccumulation of arsenic by Brassica juncea (L.) in Bangladesh

The phytoaccummulation of arsenic by Brassica juncea (L.) was investigated for varying concentrations selected within the range that is evident in Bangladeshi soil. B. juncea (Rai and BARI-11) was grown in the hydroponic media under greenhouse condition with different concentrations (0.5, 1.0, 15, 30, 50 and 100 ppm) of sodium arsenite. Inductively coupled plasma atomic emission spectroscopy (ICP-AES) was used to analyze the data. Mapping of potential area of phytoaccumulation of arsenic by B. juncea was done using Geographic information system (GIS). Arsenic was detected at lower concentrations (0.5 and 1.0 ppm) only at root system of the plant. For higher concentrations (15, 30, and 50 ppm) arsenic was detected both in the root and shoot systems. The results suggested that at 15 and 50 ppm uptake was higher compared to 30 ppm. For 100 ppm of arsenic no plant growth was observed. In Bangladesh, where concentration of arsenic is at lower level and present only at rooting zone, B. juncea may be used for phytoaccumulation of arsenic keeping usual agronomic practices. However, for higher concentrations, B. juncea can be regarded as a good accumulator of arsenic where uptake of arsenic was up to 1% of total biomass of the plant.     

Biochemical and molecular identification of Xanthomonas translucens pv. undulosa causing bacterial leaf streak of wheat in Punjab,Pakistan

Xanthomonas translucens pv. undulosa, (Xtu.), causal agent of Bacterial leaf streak (BLS) of wheat, was characterised through pathogencity, hypersensitivity, biochemical and molecular assays. Fifty symptomatic leaves of wheat were collected from eight agro-ecological zones of Punjab out of which 25 were isolated and purified. Maximum incidence and severity in Faisalabad were followed by Multan and Rahim Yar Khan. The pathogen isolated from diseased leaves was identified on the basis of colonies pattern, colour, biochemical and pathogencity test as X. translucens pv. undulosa and confirmed its pathogencity through pathogencity test. For molecular characterization, the bacterial 16S–23S rDNA spacer fragments were amplified by PCR with conserved primers (C1 and C2) and then in combination with specific primers (T1 & T2). 300?bp product amplified by C1 and C2 primer pair confirmed the presence of Xanthomonas, while specific primers T1 and T2 amplified a product of 200?bp, confirmed the presence of X. translucens pv. undulosa. This work will be quite helpful for wheat pathologist and breeders for future management strategy for this disease.     

Correlation between Li Plating Behavior and Surface Characteristics of Carbon Matrix toward Stable Li Metal Anodes

Carbonaceous materials are widely employed to host Li for stable and safe Li metal batteries while relatively little effort is devoted to tailoring the surface properties of carbon to facilitate uniform Li plating. Herein, the correlation between Li plating behavior and the surface characteristics of electrospun porous carbon nanofibers (PCNFs) is systemically elucidated through experiments and theoretical calculations. It is revealed that the neat carbon surface suffers from severe lattice mismatch with Li metal, hindering uniform Li plating. In contrast, open pores created on the PCNF surface serve as active sites for controlled initial nucleation of Li. The introduction of oxygenated functional groups further facilitates the nucleation of Li on PCNFs through the largely reduced nucleation energy barrier. The Li film uniformly deposited on PCNFs enables efficient use of the whole carbon surface, giving rise to enhanced cyclic stability of the electrode. When used as an anode in lithium–sulfur batteries, the modified electrode delivers an excellent energy density of 385 Wh kg^?1 after 100 cycles. The fundamental correlation established in this study is universal to all types of carbonaceous materials and sheds new light on the rational design of high‐performance Li metal anodes by controlling the initial Li nucleation.     

Phytoremediation of landfill leachate waste contaminants through floating bed technique using water hyacinth and water lettuce

Abstract

In the present study, the effectiveness of water hyacinth and water lettuce was tested for the phytoremediation of landfill leachate for the period of 15?days. Fifteen plastic containers were used in experimental setup where aquatic plants were fitted as a floating bed with the help of thermo-pole sheet. It was observed that both plants significantly (p?<?0.05/p?<?0.01/p?<?0.001) reduce the physicochemical parameters pH, TDS, BOD, COD and heavy metals like Zn, Pb, Fe, Cu and Ni from landfill leachate. Maximum reduction in these parameters was obtained at 50% and 75% landfill leachate treatment and their removal rate gradually increased from day 3 to day 15 of the experiment. The maximum removal rate for heavy metals such as for Zn (80–90%), Fe (83–87%) and Pb (76–84%) was attained by Eichhornia crassipes and Pistia stratiotes. Value of bioconcentration and translocation factor was less than 1 which indicates the low transport of heavy metals from roots to the above-ground parts of the plants. Both these plants accumulate heavy metals inside their body without showing much reduction in growth and showing tolerance to all the present metals. Therefore, results obtained from the study suggest that these aquatic plants are suitable candidate for the removal of pollution load from landfill leachate.     

Physical and chemical mutagens improved Sporotrichum thermophile,strain ST20 for enhanced Phytase activity

ObjectivePhosphorous is an essential micronutrient of plants and involved in critical biological functions. In nature, phosphorous is mostly present in immobilized inorganic mineral and in the fixed organic form including phytic acid and phosphoesteric compounds. However, the bioavailability of bound phosphorous could be enhanced by the use of phosphate solubilizing microorganisms such as bacteria and fungi. The phytases are widespread in an environment and have been isolated from different sources comprising bacteria and fungi.MethodologyIn current studies, we show the successful use of gamma rays and EMS (Ethyl Methane Sulphonate) mutagenesis for enhanced activity of phytases in a fungal strain Sporotrichum thermophile.ResultsWe report an improved strain ST2 that could produce a clear halo zone around the colony, up to 24 mm. The maximum enzymatic activity was found of 382 U/mL on pH 5.5. However, the phytase activity was improved to 387 U/ml at 45 °C. We also report that the mutants produced through EMS showed the greater potential for phytase production.ConclusionThe current study highlights the potential of EMS mutagenesis for strain improvement over physical mutagens.     

An integrative approach combining ion mobility mass spectrometry,X-ray crystallography,and nuclear magnetic resonance spectroscopy to study the conformational dynamics of α1-antitrypsin upon ligand binding

Native mass spectrometry (MS) methods permit the study of multiple protein species within solution equilibria, whereas ion mobility (IM)-MS can report on conformational behavior of specific states. We used IM-MS to study a conformationally labile protein (α₁-antitrypsin) that undergoes pathological polymerization in the context of point mutations. The folded, native state of the Z-variant remains highly polymerogenic in physiological conditions despite only minor thermodynamic destabilization relative to the wild-type variant. Various data implicate kinetic instability (conformational lability within a native state ensemble) as the basis of Z α₁-antitrypsin polymerogenicity. We show the ability of IM-MS to track such disease-relevant conformational behavior in detail by studying the effects of peptide binding on α₁-antitrypsin conformation and dynamics. IM-MS is, therefore, an ideal platform for the screening of compounds that result in therapeutically beneficial kinetic stabilization of native α₁-antitrypsin. Our findings are confirmed with high-resolution X-ray crystallographic and nuclear magnetic resonance spectroscopic studies of the same event, which together dissect structural changes from dynamic effects caused by peptide binding at a residue-specific level. IM-MS methods, therefore, have great potential for further study of biologically relevant thermodynamic and kinetic instability of proteins and provide rapid and multidimensional characterization of ligand interactions of therapeutic interest.PDB Code(s): 4PYW