Direct binding and characterization of laccase onto iron oxide nanoparticles.

Iron oxide nanoparticles (IONPs) exhibit unique magnetic properties and possess a high surface-to-volume ratio, making them ideal candidates for the conjugation of substances, including enzymes. Laccase (EC 1.10.3.2), an oxidative enzyme with diverse applications, presents an opportunity for enhancing stability and reusability through innovative immobilization techniques, thus reducing overall process costs. In this study, we employed a direct binding procedure via carbodiimide activation to conjugate laccase onto IONPs synthesized using thermal chemical coprecipitation. Stabilization of the nanoparticles was achieved using thioglycerol and polyvinyl alcohol (PVA) as capping agents. Characterization of the synthesized nanoparticles was conducted using UV-spectroscopy, Fourier transform infrared spectroscopy (FTIR), x-ray diffraction, scanning electron microscopy, and energy dispersive x-ray spectroscopy. FTIR spectroscopy analysis confirmed successful laccase binding to magnetic nanoparticles, with binding efficiencies of 90.65% and 73.02% observed for thioglycerol and PVA capped IONPs, respectively. Furthermore, the conjugated enzyme exhibited remarkable stability, retaining nearly 50% of its initial activity after 20 reuse cycles. This research demonstrates that immobilizing laccase onto IONPs enhances its activity, stability, and reusability, with the potential for significant cost savings and expanded applications in various fields.

The Effects of Yoga Practice on Lung Function and sIL-2R Biomarkers in Individuals Working and Living in the Lonavala Industrial Area: A Randomized Controlled Trial.

Context: Pollution-related health hazards are very common among people living and/or working in industrial areas, particularly near industries and metro cities. These air pollutants contribute to allergens, increase inflammation, and affect lung function. Aim: The present study aimed to examine the effect of yoga training on lung functions and inflammation in terms of soluble interleukin-2 receptor (sIL-2R) in people working and living in a polluted area. Settings and Design: This is a randomized controlled interventional pilot study. Methods and Materials: Forty-eight male volunteers from the industrial area, aged 20-50 years, were randomly assigned to the experimental group and the control group. Each group comprises 24 study participants. Lung function and sIL-2R were studied at the baseline and post-yoga intervention period of 16 weeks. Statistical Analysis Used: Study data were analyzed using descriptive methods, a one-tailed t-test, a paired t-test, and an independent t-test. Result: Study results showed directional and significant improvements in forced vital capacity, forced expiratory volume in the first second, and peak expiratory flow rate compared to the control group participants. The results pertaining to sIL-2R showed a directional and significant decrease in the experimental group compared to the control group. Conclusion: The present study showed that yoga helps to promote better health, improve lung function and reduce inflammation among people residing in polluted environments.

Abundance of Phasi-Charoen-like virus in Aedes aegypti mosquito populations in different states of India.

Mosquitoes are known to harbor a large number of insect specific viruses (ISV) in addition to viruses of public health importance. These ISVs are highly species specific and are non-pathogenic to humans or domestic animals. However, there is a potential threat of these ISVs evolving into human pathogens by genome alterations. Some ISVs are known to modulate replication of pathogenic viruses by altering the susceptibility of vector mosquitoes to pathogenic viruses, thereby either inhibiting or enhancing transmission of the latter. In the present study, we report predominance of Phasi Charoen-like virus (PCLV, Family: Phenuviridae) contributing to >60% of the total reads in Aedes aegypti mosquitoes collected from Pune district of Maharashtra state using next generation sequencing based metagenomic analysis of viromes. Similar results were also obtained with mosquitoes from Assam, Tamil Nadu and Karnataka states of India. Comparison of Pune mosquito sequences with PCLV Rio (Brazil) isolate showed 98.90%, 99.027% and 98.88% homologies in the S, M and L segments respectively indicating less genetic heterogeneity of PCLV. The study also demonstrated occurrence of transovarial transmission as seen by detection of PCLV in eggs, larvae, pupae and male mosquitoes. Ae. aegypti mosquitoes collected from Pune also showed a large number of reads for viruses belonging to Baculoviridae, Rhabdoviridae, Genomoviridae and Bunyaviridae families. The role of PCLV in the replication of dengue and chikungunya virus is yet not clear. It warrants further studies to know the significance of PCLV and other ISVs on the replication and transmission of Ae. aegypti borne pathogenic viruses, especially in the absence of prophylactics or therapeutics.

Phylogenetic classification of the whole-genome sequences of SARS-CoV-2 from India & evolutionary trends.

BACKGROUND & OBJECTIVES: Several phylogenetic classification systems have been devised to trace the viral lineages of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, inconsistency in the nomenclature limits uniformity in its epidemiological understanding. This study provides an integration of existing classifications and describes evolutionary trends of the SARS-CoV-2 strains circulating in India. METHODS: The whole genomes of 330 SARS-CoV-2 samples were sequenced using next-generation sequencing (NGS). Phylogenetic and sequence analysis of a total of 3014 Indian SARS-CoV-2 sequences from 20 different States/Union Territories (January to September 2020) from the Global Initiative on Sharing All Influenza Data (GISAID) database was performed to observe the clustering of Nextstrain and Phylogenetic Assignment of Named Global Outbreak LINeages (Pangolin) lineages with the GISAID clades. The identification of mutational sites under selection pressure was performed using Mixed Effects Model of Evolution and Single-Likelihood Ancestor Counting methods available in the Datamonkey server. RESULTS: Temporal data of the Indian SARS-CoV-2 genomes revealed that except for Uttarakhand, West Bengal and Haryana that showed the circulation of GISAID clade O even after July 2020, the rest of the States showed a complete switch to GR/GH clades. Pangolin lineages B.1.1.8 and B.1.113 identified within GR and GH clades, respectively, were noted to be indigenous evolutions. Sites identified to be under positive selection pressure within these clades were found to occur majorly in the non-structural proteins coded by ORF1a and ORF1b. INTERPRETATION & CONCLUSIONS: This study interpreted the geographical and temporal dominance of SARS-CoV-2 strains in India over a period of nine months based on the GISAID classification. An integration of the GISAID, Nextstrain and Pangolin classifications is also provided. The emergence of new lineages B.1.1.8 and B.1.113 was indicative of host-specific evolution of the SARS-CoV-2 strains in India. The hotspot mutations such as those driven by positive selection need to be further characterized.

Comparative Proteomics Unravels the Differences in Salt Stress Response of Own-Rooted and 110R-Grafted Thompson Seedless Grapevines.

Thompson Seedless, a commonly grown table grape variety, is sensitive to salinity when grown on its own roots, and therefore, it is frequently grafted onto salinity-tolerant wild grapevine rootstocks. Rising soil salinity is a growing concern in irrigated agricultural systems. The accumulation of salts near the root zone severely hampers plant growth, leading to a decrease in the productive lifespan of grapevine and causing heavy yield losses to the farmer. In the present study, we investigated the differences in response to salinity between own-rooted Thompson Seedless (TSOR) and 110R-grafted Thompson Seedless (TS110R) grapevines, wherein 110R is reported to be a salt-tolerant rootstock. The grapevines were subjected to salt stress by treating them with a 150 mM NaCl solution. The stress-induced changes in protein abundance were investigated using a label-free shotgun proteomics approach at three time-points viz. 6 h, 48 h, and 7 days of salt treatment. A total of 2793 proteins were identified, of which 246 were differentially abundant at various time-points in TSOR and TS110R vines. The abundance of proteins involved in several biological processes such as photosynthesis, amino acid metabolism, translation, chlorophyll biosynthesis, and generation of precursor metabolites was significantly affected by salt stress in both the vines but at different stages of stress. The results revealed that TSOR vines responded fervently to salt stress, while TS110R vines adopted a preventive approach. The findings of this study add to the knowledge of salinity response in woody and grafted plants and hence open the scope for further studies on salt stress-specific differences induced by grafting.

Prospective of Microbial Exopolysaccharide for Heavy Metal Exclusion.

Metals as a resource are depleting, and on another side, it fetches serious environmental pollution causing a threat to human health and ecosystem. The heavy metal accumulation due to anthropogenic activities results in toxicological manifestation. The traditional methods of remediation are not cost effective, efficient, and ecofriendly which necessitate and motivate towards the safe, effective, and ecofriendly biological methods. The increasing presence of heavy metals in the microbial habitat compels the microbes to develop the ability to tolerate or resist the presence of heavy metals. Exopolysaccharide (EPS) production is one of the strategies of microbes to fight against metal stress. EPS is a microbial biopolymer which is generally produced under stress from harsh environment and nutrition conditions. EPSs are cell-associated or secreted outside the cell and comprised organic macromolecules such as polysaccharides, proteins, and phospholipids in addition to some non-polymeric molecules. EPSs work as competent biosorbents with an anionic reactant group that effectively sequesters cationic heavy metals by electrostatic interactions. The present paper summarizes the EPSs with its types, role, and biosynthesis and an endeavor to elucidate the interaction mechanism of EPSs with heavy metal with supportive and distinctive applications for heavy metal exclusion. The review concluded with the current challenges and future prospects to make the EPS an efficient biosorbent.

Effect of different carbon sources on morphology and silver accumulation in Cochliobolus lunatus.

The morphology of filamentous fungi plays very important role in uptake of metabolites and enzyme production. A filamentous fungus may be fibrous, hyphae, pellets, clumps, etc. Cochliobolus lunatus is a fungus which has previously been reported for silver accumulation and nanoparticles formation. The present study investigated the role of various carbon sources on morphology, biochemical profile, silver accumulation, and biosynthesis of silver nanoparticles by fungal strain C. lunatus. In this investigation, effect of different carbon sources was studied on morphology of C. lunatus and its silver accumulating ability. As a result of different carbon sources like carboxymethyl cellulose (CMC), pectin, starch, agar, sucrose, and mannitol, the organism showed three kinds of morphologies like homogenous smooth branched clumps, tough short fibrous filaments, and tough pellets, as well as silver accumulating ability. Atomic absorption spectroscopy (AAS) studies showed maximum uptake of Ag(+): 87.44 ± 0.23 and 82.57 ± 0.19 % in pectin- and CMC-grown biomass, respectively. The crystalline nature of silver nanoparticles (AgNPs) was confirmed by X-ray diffraction studies. Transmission electron microscopy (TEM) micrographs of silver nanoparticles confirmed size ranging from 5 to 38 nm.

Biochemical, structural and functional diversity between two digestive α-amylases from Helicoverpa armigera.

BACKGROUND: Helicoverpa armigera (Lepidoptera) feeds on various plants using diverse digestive enzymes as one of the survival tool-kit. The aim of the present study was to understand biochemical properties of recombinant α-amylases of H. armigera viz., HaAmy1 and HaAmy2. METHODS: The open reading frames of HaAmy1 and HaAmy2 were cloned in Pichia pastoris and expressed heterologously. Purified recombinant enzymes were characterized for their biochemical and biophysical attributes using established methods. RESULTS: Sequence alignment and homology modeling showed that HaAmy1 and HaAmy2 were conserved in their amino acid sequences and structures. HaAmy1 and HaAmy2 showed optimum activity at 60°C; however, they differed in their optimum pH. Furthermore, HaAmy2 showed higher affinity for starch and amylopectin whereas HaAmy1 had higher catalytic efficiency. HaAmy1 and HaAmy2 were inhibited to the same magnitude by a synthetic amylase inhibitor (acarbose) while wheat amylase inhibitor showed about 2-fold higher inhibition of HaAmy1 than HaAmy2 at pH7 while 6-fold difference at pH11. Interactions of HaAmy1 and HaAmy2 with wheat amylase inhibitor revealed 2:1 stoichiometric ratio and much more complex interaction with HaAmy1. CONCLUSIONS: The diversity of amylases in perspective of their biochemical and biophysical properties, and their differential interactions with amylase inhibitors signify the potential role of these enzymes in adaptation of H. armigera on diverse plant diets. GENERAL SIGNIFICANCE: Characterization of digestive enzymes of H. armigera provides the molecular basis for the polyphagous nature and thus could assist in designing future strategies for the insect control.