Development of Pectinase Based Nanocatalyst by Immobilization of Pectinase on Magnetic Iron Oxide Nanoparticles Using Glutaraldehyde as Crosslinking Agent.

To increase its operational stability and ongoing reusability, B. subtilis pectinase was immobilized on iron oxide nanocarrier. Through co-precipitation, magnetic iron oxide nanoparticles were synthesized. Scanning electron microscopy (SEM) and energy dispersive electron microscopy (EDEX) were used to analyze the nanoparticles. Pectinase was immobilized using glutaraldehyde as a crosslinking agent on iron oxide nanocarrier. In comparison to free pectinase, immobilized pectinase demonstrated higher enzymatic activity at a variety of temperatures and pH levels. Immobilization also boosted pectinase's catalytic stability. After 120 h of pre-incubation at 50 °C, immobilized pectinase maintained more than 90% of its initial activity due to the iron oxide nanocarrier, which improved the thermal stability of pectinase at various temperatures. Following 15 repetitions of enzymatic reactions, immobilized pectinase still exhibited 90% of its initial activity. According to the results, pectinase's catalytic capabilities were enhanced by its immobilization on iron oxide nanocarrier, making it economically suitable for industrial use.

Characterization of proteome wide antigenic epitopes to design proteins specific and proteome-wide ensemble vaccines against heartland virus using structural vaccinology and immune simulation approaches.

Heartland virus is a single-stranded negative-sense RNA virus that infects humans and causes lethargy, myalgia, headaches, nausea, diarrhea, weight loss, arthralgia, loss of appetite, leukopenia, and easy bruising due thrombocytopenia. The unavailability of antiviral drugs for HRTV infection is a major obstacle to treat this infection, therefore supportive care management is adopted in the case of a severe ailment. In this scientific study, proteins specific and proteome-wide Helper T-cell (HTL), linear B cell, and cytotoxic T-cell (CTL) epitopes mapping joined together with suitable linkers to design multi-epitopes subunit vaccine (MEVC). The constructed four vaccines from nucleocapsid protein, replicase, glycoprotein and finally whole proteome-wide constructs demonstrated stronger antigenic and non-allergenic behavior. Physiochemical properties evaluation also reported easy and efficient expression and downstream analysis of the constructs. Molecular docking of these constructs with toll-like receptor 7 (TLR7) revealed good binding and further validation based on MM/GBSA also demonstrated stronger interaction between the vaccine constructs and TLR7. Moreover, in silico cloning reported CAI value of 0.96 for each construct and excellent GC contents percentage required for experimental analysis. Furthermore, immune simulation-based immune response surveillance revealed that upon the injection of antigen the primary and secondary antibodies were produced between 5 and 15 days, and a more robust neutralization of the antigen by the proteome-wide vaccine construct was observed. This research could pave the way for the development of dynamic and efficient vaccines that contain a unique mix of numerous HRTV derived antigenic peptides to control HRTV infection.

Assessment of Pb and Ni and potential health risks associated with the consumption of vegetables grown on the roadside soils in District Swat, Khyber Pakhtunkhwa, Pakistan.

Vegetables cultivated near roads absorb toxic metals from polluted soil, which enter the human body through the food chain and cause serious health problems to humans. The present study investigated the concentration of lead (Pb) and nickel (Ni) in soils and vegetables grown along the roadside of District Swat, Pakistan, and the health risks associated with the consumption of the tested vegetables. In results, Pb concentration was higher in plants located at the distance between 0-10 m away from the roadside than the WHO permissible limit. In such plants, Pb concentration was higher than Ni. Rumex dentatus contained the highest concentration of Pb (75.63 mg kg-1 DW) among the tested vegetables while Ni concentration (27.57 mg kg-1 DW) was highest in Trachyspermum ammi as compared to other plants. Concentration and accumulation of both the metals decreased in soil and plants with increasing distance from the road. Similarly, target hazard quotient values noted for Pb (up to 3.37) were greater than unity, which shows that there is a potential risk associated with the consumption of tested vegetables near the road. Moreover, the values of target cancer risk (up to 0.8413) were greater than 0.0001, which shows that there is a risk of cancer with the consumption of tested vegetables. In conclusion, the consumption of tested vegetables was very dangerous as it may lead to higher risks of cancer. Strict regulatory control is recommended on the cultivation of these vegetables along the roadside to avoid any contamination due to roadside exhaust.

Assessment of health and ecological risks of heavy metal contamination: a case study of agricultural soils in Thall, Dir-Kohistan.

Heavy metal (HM) contamination in agricultural soils has been a significant health concern worldwide due to their persistent and non-biodegradable nature and biomagnification to higher trophic levels. The present study was conducted to assess Cd and Pb concentrations in soil samples collected from potato-growing areas in Thall, Dir-Kohistan, and to determine their associated health and ecological risks. The contamination factor calculated for Cd (0 to 1.74) and Pb (0 to 0.91) showed their moderate to low contamination in the study area. Geo-accumulation indices of Cd and Pb were less than 1, indicating moderate soil pollution of these HM in the study area. However, the ecological risk factor (Eri) of Cd was greater than 40 for only three soil samples, indicating the moderate potential of ecological risks of respective soils. The principal component analysis (PCA) and Pearson correlation suggested that the contamination in different soils was lithogenic followed by anthropogenic activities. The hazard quotient (HQ) in children and adults was found in the following order: dermal > ingestion > inhalation. Moreover, the values of HQ through various exposure routes were higher in children compared to adults, which showed that adults were at a lower level of risk associated with HM contamination. The results of the present study can serve as baseline data for government agencies related to environmental protection, which could devise policies to minimize Cd and Pb contamination in the agricultural soils.

Sources, impacts, factors affecting Cr uptake in plants, and mechanisms behind phytoremediation of Cr-contaminated soils.

Chromium (Cr) is released into the environment through anthropogenic activities and has gained significant attention in the recent decade as environmental pollution. Its contamination has adverse effects on human health and the environment e.g. decreases soil fertility, alters microbial activity, and reduces plant growth. It can occur in different oxidation states, with Cr(VI) being the most toxic form. Cr contamination is a significant environmental and health issue, and phytoremediation offers a promising technology for remediating Cr-contaminated soils. Globally, over 400 hyperaccumulator plant species from 45 families have been identified which have the potential to remediate Cr-contaminated soils through phytoremediation. Phytoremediation can be achieved through various mechanisms, such as phytoextraction, phytovolatilization, phytodegradation, phytostabilization, phytostimulation, and rhizofiltration. Understanding the sources and impacts of Cr contamination, as well as the factors affecting Cr uptake in plants and remediation techniques such as phytoremediation and mechanisms behind it, is crucial for the development of effective phytoremediation strategies. Overall, phytoremediation offers a cost-effective and sustainable solution to the problem of Cr pollution. Further research is needed to identify plant species that are more efficient at accumulating Cr and to optimize phytoremediation methods for specific environmental conditions. With continued research and development, phytoremediation has the potential to become a widely adopted technique for the remediation of heavy metal-contaminated soils.

Plant Growth Regulators with a Balanced Supply of Nutrients Enhance the Phytoextraction Efficiency of Parthenium hysterophorus for Cadmium in Contaminated Soil.

Heavy metal contamination in soil, such as cadmium (Cd), poses a serious threat to global food security and human health. It must be managed using environmentally friendly and cost-effective technologies. Plants with high resistance to Cd stress and high biomass production could be potential candidates for the phytoremediation of Cd-contaminated soils to improve Cd phytoextraction. In this regard, the present study was carried out to determine the effect of gibberellic acid (GA3), indole acetic acid (IAA), and fertilizers (N, P, and K) on Parthenium hysterophorus growth and biomass production as well as Cd phytoextraction capabilities. A pot experiment was conducted with various combinations of PGRs and fertilizers, with treatments arranged in five replicates using a completely randomized design. After harvesting, each plant was divided into various parts such as stems, roots, and leaves, and different growth, physiological, and biochemical parameters were recorded. Results showed that under Cd stress, growth, physiological, and biochemical parameters were all significantly decreased. With the combined application of plant growth regulators (GA3 and IAA) and nutrients, Cd stress was alleviated and all parameters significantly improved. In comparison to the control treatment, the combined application of N + P + K + GA3 + IAA resulted in the highest fresh and dry biomass production of the root (12.31 and 5.11 g pot-1), shoot (19. 69 and 6.99 g pot-1), leaves (16.56 and 7.09 g pot-1), and entire plant (48.56 and 19.19 g pot-1). Similarly, the same treatment resulted in higher chlorophyll a and b and total chlorophyll contents under Cd stress, which were 2.19, 2.03, and 3.21 times higher than the control, which was Cd stress without any treatment. The combination of N + P + K + GA3 + IAA also resulted in the highest proline and phenolic contents. In the case of different enzyme activities, the combined application of N + P + K + GA3 + IAA under Cd stress led to a high increase in catalase (2.5 times), superoxide (3.5 times), and peroxidase (3.7 times) compared to the control. With the combined application of N+ P+ K + GA3 + IAA, the maximum values of BCF (8.25), BAC (2.6), and RF (5.14%) were measured for phytoextraction potential. On the basis of these findings, it is concluded that P. hysterophorus has a high potential to grow, produce the most biomass, and act as a Cd hyperaccumulator in Cd-contaminated soil.

Plant growth regulators and EDTA improve phytoremediation potential and antioxidant response of Dysphania ambrosioides (L.) Mosyakin & Clemants in a Cd-spiked soil.

Soil pollution due to potentially toxic elements is a worldwide challenge for health and food security. Chelate-assisted phytoextraction along with the application of plant growth regulators (PGRs) could increase the phytoremediation efficiency of metal-contaminated soils. The present study was conducted to investigate the effect of different PGRs [Gibberellic acid (GA3) and indole acetic acid (IAA)] and synthetic chelator (EDTA) on growth parameters and Cd phytoextraction potential of Dysphania ambrosioides (L.) Mosyakin & Clemants grown under Cd-spiked soil. GA3 (10-7 M) and IAA (10-5 M) were applied four times with an interval of 10 days through a foliar spray, while EDTA (40 mg kg-1 soil) was once added to the soil. The results showed that Cd stress significantly decreased fresh biomass, dry biomass, total water contents, and photosynthetic pigments as compared to control. Application of PGRs significantly enhanced plant growth and Cd phytoextraction. The combined application of GA3 and IAA with EDTA significantly increased Cd accumulation (6.72 mg pot-1 dry biomass) and bioconcentration factor (15.21) as compared to C1 (Cd only). The same treatment significantly increased chlorophyll, proline, phenolic contents, and antioxidant activities (CAT, SOD, and POD) while MDA contents were reduced. In roots, Cd accumulation showed a statistically significant and positive correlation with proline, phenolics, fresh biomass, and dry biomass. Similarly, Cd accumulation showed a positive correlation with antioxidant enzyme activities in leaves. D. ambrosioides showed hyperaccumulation potential for Cd, based on bioconcentration factor (BCF) > 1. In conclusion, exogenous application of GA3 and IAA reduces Cd stress while EDTA application enhances Cd phytoextraction and ultimately the phytoremediation potential of D. ambrosioides.

Green synthesis of Zno@GO nanocomposite and its' efficient antibacterial activity.

Nanotechnologyapplications in the field of biomedicine like drug delivery, cell labeling, and bacterial inhibition are growing . New nano-materials having less toxicity and excellent antibacterial activity attract research interest. In the current study, while taking advantage of green synthesis we have decorated zinc oxide on the surface of grephene oxide forming Zno@GO nanocomposite. The Transmission electron microscopy (TEM) study showed successfully synthesized trigonal small sizes ZnO on the surface of GO nanosheets. The as-synthesized ZnO@GO was used against MDR gram-negative pathogen E-coli (BL21 DE3) and showed excellent antibacterial activity killing about 95 % toxic bacteria within 5 h due to electrostatic interaction between cell membrane of E. coli (BL21 DE3) and ZnO@GO complex. Hence the nano composite subsequently penetrated into the cytoplasm by damaging the cell membrane of bacteria, as a result production of ROS into the cytoplasm led to imbalance of metabolic system in the cell. Moreover, the cell membrane damage of gram-negative bacteria verified through zeta potential and propidium iodide (PI) study. Thus, our study develops a way to solve the challenge of efficient design of a drug delivery system for dissolution enhancement according to the need for required drug release.

Genetic characterization of structural and open reading Fram-8 proteins of SARS-CoV-2 isolates from different countries.

Since December 2019, a severe pandemic of pneumonia, COVID-19 associated with a novel coronavirus (SARS-CoV-2), have emerged in Wuhan, China and spreading throughout the world. As RNA viruses have a high mutation rate therefore we wanted to identify whether this virus is also prone to mutations. For this reason we selected four major structural (Spike protein (S), Envelope protein (E), Membrane glycoprotein (M), Nucleocapsid phosphoprotein (N)) and ORF8 protein of 100 different SARS-CoV-2 isolates of fifteen countries from NCBI database and compared these to the reference sequence, Wuhan NC_045512.2, which was the first isolate of SARS-CoV-2 that was sequenced. By multiple sequence alignment of amino acids, we observed substitutions and deletion in S protein at 13 different sites in the isolates of five countries (China, USA, Finland, India and Australia) as compared to the reference sequence. Similarly, alignment of N protein revealed substitutions at three different sites in isolates of China, Spain and Japan. M protein exhibits substitution only in one isolates from USA, however, no mutation was observed in E protein of any isolate. Interestingly, in ORF8 substitution of Leucine, a nonpolar to Serine a polar amino acid at same position (aa84 L to S) in 23 isolates of five countries i.e. China, USA, Spain, Taiwan and India were observed, which may affect the conformation of peptides. Thus, we observed several mutations in the isolates thereafter the first sequencing of SARS-CoV-2 isolate, NC_045512.2, which suggested that this virus might be a threat to the whole world and therefore further studies are needed to characterize how these mutations in different proteins affect the functionality and pathogenesis of SARS-CoV-2.

Potassium and zinc increase tolerance to salt stress in wheat (Triticum aestivum L.).

Potassium and zinc are essential elements in plant growth and metabolism and plays a vital role in salt stress tolerance. To investigate the physiological mechanism of salt stress tolerance, a pot experiment was conducted. Potassium and zinc significantly minimize the oxidative stress and increase root, shoot and spike length in wheat varieties. Fresh and dry biomass were significantly increased by potassium followed by zinc as compared to control C. The photosynthetic pigment and osmolyte regulator (proline, total phenolic, and total carbohydrate) were significantly enhanced by potassium and zinc. Salt stress increases MDA content in wheat varieties while potassium and zinc counteract the adverse effect of salinity and significantly increased membrane stability index. Salt stress decreases the activities of antioxidant enzymes (superoxide dismutase, catalase and ascorbate peroxidase) while the exogenous application of potassium and zinc significantly enhanced the activities of these enzymes. A significant positive correlation was found of spike length with proline (R2 = 0.966 ∗∗∗), phenolic (R2 = 0.741∗) and chlorophyll (R2 = 0.853∗∗). The MDA content showed significant negative correlation (R2 = 0.983∗∗∗) with MSI. It is concluded that potassium and zinc reduced toxic effect of salinity while its combine application showed synergetic effect and significantly enhanced salt tolerance.

Bio-fabrication of catalytic platinum nanoparticles and their in vitro efficacy against lungs cancer cells line (A549).

Platinum based drugs are considered as effective agents against various types of carcinoma; however, the severe toxicity associated with the chemically prepared platinum complexes limit their practical applications. Similarly, water pollution caused by various organic moieties is another serious health problem worldwide. Hence, an intense need exists to develop new, effective and biocompatible materials with catalytic and biomedical applications. In the present contribution, we prepared platinum nanoparticles (PtNPs) by a green route using phytochemicals as a source of reducing and stabilizing agents. Well dispersed and crystalline PtNPs of spherical shapes were prepared and characterized. The bio-fabricated PtNPs were used as catalyst and anticancer agents. Catalytic performance of the PtNPs showed that 84% of the methylene blue can be reduced in 32min under visible light irradiation (K=0.078min-1). Similarly the catalytic conversion of 4-nitrophenol to 4-aminophenol was achieved in <20min (K=0.124min-1). The in vitro anticancer study revealed that biogenic PtNPs are the efficient nano-agents possessing strong anticancer activity against the lungs cancer cells line (A549). Interestingly, the as prepared PtNPs were well tolerated by normal human cells, and therefore, could be effective and biocompatible agents in the treatment of different cancer cells.

Enhanced phytoremediation of cadmium polluted water through two aquatic plants Veronica anagallis-aquatica and Epilobium laxum.

Toxic metal-contaminated water is a major threat to sustainable agriculture and environment. Plants have the natural ability to absorb and concentrate essential elements in its tissues from water solution, and this ability of plants can be exploited to remove heavy/toxic metals from the contaminated water. For this purpose, two plants Veronica anagallis-aquatica and Epilobium laxum were hydroponically studied. The effect of different fertilizers (NPK) and plant growth regulators (GA3 and IAA) were evaluated on growth, biomass, free proline, phenolics, and chlorophyll contents, and their role in Cd phytoaccumulation was investigated. Results showed that in both plants, fertilizer addition to media (treatment T4) produced the highest significant increase in growth, biomass (fresh and dry), cadmium concentration, proline, phenolics, and chlorophyll concentrations. The significant effect of GA3 in combination with NPK foliar spray (treatment T12) was observed on most of the growth parameters, Cd concentration, and proline and phenolic contents of the plants. The free proline and total phenolics showed positive correlation with cadmium concentration within plant tissues. Proline showed significantly positive correlation with phenolic contents of root and shoot. Veronica plant demonstrated the hyperaccumulator potential for cadmium as bioconcentration factor (BCF >1) which was much higher than 1, while Epilobium plant showed non-hyperaccumulator potential. It is recommended for further study to investigate the role of Veronica plant for other metals and to study the role of phenolics and proline contents in heavy metal phytoextraction by various plant species.

Determination of optimal toxic concentration and accumulation of cadmium in broiler chicks.

Cadmium is considered one of the most toxic, non biodegradable heavy metal for the human and animals. The purpose of the present study was to investigate the changes in biochemical parameters of blood and accumulation of cadmium in various tissue caused by various levels of dietary cadmium chloride (CdCl2) in broiler chicks. CdCl2 was administered through drinking water to broiler chicks. In spectral analysis, CdCl2 treatment caused a significant increase in Glutamate pyruvate transaminase (GPT), creatinine and uric acid levels in all treated groups. Intriguingly, the GPT, creatinine, and uric acid levels were significantly higher at 75 mg/kg as compared to the groups treated with high doses (100, 125 and 150 mg/kg) of CdCl2. Atomic Absorption Spectrophotometer (AAS) was used for the determination of Cd accumulation in kidney, liver and Breast muscles. AAS analysis revealed that Cd accumulation is increased in breast muscles as compared to liver and kidney at higher doses of Cd than 75 mg/kg.