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The cultivation of forage crops on wastewater-irrigated soils, while common in many developing countries, poses significant risks due to heavy metal pollution, particularly Lead (Pb) and Nickel (Ni). This practice, aimed at addressing water scarcity challenges and providing affordable irrigation, was investigated for its ecological and human health implications across three diverse sites (site A, site B, and site C). Our study unveiled increases in Pb concentrations in contaminated soil, cultivated with Sesbania bispinosa showing the highest Pb accumulation. The Ni concentrations ranged from 5.34 to 10.43 across all forage crop samples, with S. fruticosa from site C displaying the highest Ni concentration and S. bicolor from site A exhibiting the lowest. Trace element concentrations in the specimens were determined using an atomic absorption spectrophotometer. The Pb levels in the blood, hair, and feces of farm ruminants (cows, buffaloes, and sheep) varied across the sites, with buffaloes consistently displaying the highest Pb levels. Insights into daily Pb intake by ruminant's highlighted variations influenced by plant species, animal types, and sites, with site C, the cows exhibiting the highest Health Risk Index (HRI) associated with lead exposure from consuming forage crops. Soil and forage samples showed Pb concentrations ranging from 8.003 to 12.29 mg/kg and 6.69-10.52 mg/kg, respectively, emphasizing the severe health risks associated with continuous sewage usage. Variations in Ni concentrations across animal blood, hair, and feces samples underscored the importance of monitoring Ni exposure in livestock, with sheep at site B consistently showing the highest Ni levels. These findings highlight the necessity of vigilance in monitoring trace element (Pb and Ni) exposure in forage crops and livestock, to mitigate potential health risks associated with their consumption, with variations dependent on species, site, and trace element concentrations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Purpose: Fascioliasis is a zoonotic parasitic disease spread by genus Fasciola that causes considerable health and economic losses to both humans and livestock. Pakistan is an endemic region for fascioliasis and only a few molecular studies have been conducted to describe the etiology of disease in the country., Methods: Current study involved the molecular characterization of adult fluke specimens (n = 31) through ITS-1 region of rDNA collected from four host species (sheep, goats, cattle and buffaloes) of district Sargodha. Moreover, coprology based epidemiology (n = 28620) and association of different risk factors with disease prevalence was also determined. Additionally, an overview of prevalence of fascioliasis in Pakistan, including molecular studies in the four provinces and livestock species, was also carried out., Results: The phylogenetic analysis revealed that Fasciola gigantica was the main etiological agent infecting the livestock species of this region. The coprological survey revealed an overall prevalence of 30.18% among the livestock species (sheep = 32.31%, goats = 30.67%, cattle = 31.01%, buffaloes = 25.23%). It was inferred that females were slightly more susceptible to infection (32.59%) and that the older age group had higher infection rate (34.01%) in all the host species., Conclusion: The current study suggests that fascioliasis is a disease of primary concern indicating its status as one of the prevalent diseases affecting livestock in this region. It was also concluded that both Fasciola gigantica and Fasciola hepatica existed among the infected animals in addition to presence of hybrid/intermediate form in Pakistan with F. gigantica as dominant species in Punjab province., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

One of the most pressing environmental issues is how to properly dispose of municipal solid waste (MSW), which represents both a substantial source of concern and a challenge. The current study evaluated cobalt (Co) accumulation in MSW, their uptake by different vegetables grown for two years, and related human health risks. Vegetables were grown in four different groups, such as one control (ground soil), and the remaining treatment groups (T1, T2, and T3) received varying concentrations of MSW. The analysis of Co was done through an atomic absorption spectrophotometer (AAS). Results revealed that the concentration of Co was higher in all the vegetables (n = 15) grown in soil supplemented with 75% MSW during 2nd growing year. Among all vegetables, the highest concentration of Co was observed in Solanum tuberosum at T3 during 2nd growing year. The pollution load index (PLI) value for vegetables during both growing years was more than 1 except in control soil. The findings indicated that the highest enrichment factor (EF) and hazard resilience index (HRI) value of 0.09 was present in S. tuberosum. Health index values for cobalt in the study were below 1. The HRI < 1 indicated that consumers do not face any immediate health risks. The investigation of Co concentrations in blood samples obtained from individuals residing in different areas contributes a human health perspective to the research. The findings indicate that the concentration of Co rises with an increasing proportion of MSW. While the metal levels in MSW-treated soil were not high enough to classify the soil as polluted, the results recommend that recycling MSW can substitute mineral fertilizers. Nevertheless, the presence of cobalt in MSW may directly affect soil fertility and could impact crop production and human health., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In this study, the concentrations of cadmium (Cd), iron (Fe), manganese (Mn), nickel (Ni), lead (Pb) and zinc (Zn) in plants sampled from Mt. Madra were investigated. Furthermore, the distribution characteristics and source identification of potentially toxic metals were investigated with the application of Positive Matrix Factorization (PMF) modelling. Samples of 26 different plant species were taken from Mt. Madra at elevations ranging from 177 to 1347 m using the multi-point sampling approach. The metal quantities measured by ICP-OES are the following sequences (mean ± SD) (mg/kg): Fe (974.96 ± 29.6) > Mn (111.81 ± 2.6) > Zn (27.28 ± 0.2) > Ni (2.17 ± 0.03) > Pb (0.77 ± 0.01) > Cd (0.12 ± 0.01). According to the plant samples in which the highest values were determined, the metals are as follows: Cd (Lathyrus laxiflorus, 0.401 mg/kg), Fe (Ajuga orientalis, 7621.207 mg/kg), Mn (Castanea sativa, 724.927 mg/kg), Ni (Prunella laciniata, 6.947 mg/kg), Pb (Crataegus stevenii, 3.955 mg/kg) and Zn (Prunella laciniata, 50.802 mg/kg). The results of the PMF model showed that Cd had an atmospheric transport factor originated and transported from industrial activites, Ni had a substrate factor, Fe, Mn, Pb and Zn were influenced by different anthropogenic factors., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Timely and unbiased evaluation of Autism Spectrum Disorder (ASD) is essential for providing lasting benefits to affected individuals. However, conventional ASD assessment heavily relies on subjective criteria, lacking objectivity. Recent advancements propose the integration of modern processes, including artificial intelligence-based eye-tracking technology, for early ASD assessment. Nonetheless, the current diagnostic procedures for ASD often involve specialized investigations that are both time-consuming and costly, heavily reliant on the proficiency of specialists and employed techniques. To address the pressing need for prompt, efficient, and precise ASD diagnosis, an exploration of sophisticated intelligent techniques capable of automating disease categorization was presented. This study has utilized a freely accessible dataset comprising 547 eye-tracking systems that can be used to scan pathways obtained from 328 characteristically emerging children and 219 children with autism. To counter overfitting, state-of-the-art image resampling approaches to expand the training dataset were employed. Leveraging deep learning algorithms, specifically MobileNet, VGG19, DenseNet169, and a hybrid of MobileNet-VGG19, automated classifiers, that hold promise for enhancing diagnostic precision and effectiveness, was developed. The MobileNet model demonstrated superior performance compared to existing systems, achieving an impressive accuracy of 100%, while the VGG19 model achieved 92% accuracy. These findings demonstrate the potential of eye-tracking data to aid physicians in efficiently and accurately screening for autism. Moreover, the reported results suggest that deep learning approaches outperform existing event detection algorithms, achieving a similar level of accuracy as manual coding. Users and healthcare professionals can utilize these classifiers to enhance the accuracy rate of ASD diagnosis. The development of these automated classifiers based on deep learning algorithms holds promise for enhancing the diagnostic precision and effectiveness of ASD assessment, addressing the pressing need for prompt, efficient, and precise ASD diagnosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Alsharif, Al-Adhaileh, Al-Yaari, Farhah and Khan.)

The aim of this study was to evaluate the impact of different irrigation sources on the levels of potentially toxic metals (Cd, Cr, Fe and Mn) in the edibles of citrus fruits (Citrus sinensis and Citrus limetta). The samples of fruit, soil and water were collected from two locations (fresh water irrigated-FW I and sewage water irrigated-SW II) within the city of Sargodha. The samples utilized in the study for metal analysis were prepared utilizing the wet acid digestion method. Metal determination was performed using Atomic Absorption Spectrometry (AAS). The potentially toxic metal values in the citrus samples ranged from 0.010 to 0.063, 0.015 to 0.293, 6.691 to 11.342 and 0.366 to 0.667 mg/kg for Cd, Cr, Fe and Mn, respectively. Analysis of Citrus limetta and Citrus sinensis indicated that the highest concentration of Cr, Fe and Mn is observed at the sewage water irrigation site (SW-II), whilst the minimum levels of Cr, Fe and Mn were observed at the fresh water irrigation site (FW-I). The results show that the levels of these metals in soil and fruit samples meet the acceptable guidelines outlined by USEPA and WHO. It was found that the metal pollution constitutes a potential threat to human health due to the HRI values for Cd, Cr, and Fe being above 1, despite the DIM values being below 1. Regular monitoring of vegetables irrigated with wastewater is highly recommended in order to minimise health risks to individuals., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Composting is a process of microbial degradation of organic waste and is commonly applied for waste management. This is a slow process and requires a lot of land and human resources. The present study investigated mechanical augmentation with required microbial culture for composting municipal solid waste (MSW). Thirty isolates were subjected to 16S rDNA PCR amplification and gene sequencing. The isolates' sequencing from the compost samples was processed on BLASTn. Fourteen strains were identified for further experiments. The results divulge that Empedobacter (04), Bacillus (02), Proteus (02), Lactiplantibacillus (01), Klebsiella (01), Citrobacter (01), Brevibacillus (01), E. coli (01) and one unidentified strain were growing during composting. Eleven combinations of bacterial consortium and respective additives were applied for the organic waste decomposition in the next stage, resulting in varied completion periods ranging from 3 to 14 days. Two combinations were completed within 3 days, which are considered ideal combinations for composting. The microbial consortium was significantly diverse, which is a reason for rapid biodegradation. The present study reveals that the technology will be highly feasible for municipal solid waste management in tropical/subtropical countries., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

In many countries with wastewater irrigation and intensive use of fertilizers (minerals and organics), heavy metal deposition by crops is regarded as a major environmental concern. A study was conducted to determine the impact of mineral fertilizers, cow manure, poultry manure, leaf litter, and sugarcane bagasse on soil's trace Pb content and edible parts of vegetables. It also evaluated the risk of lead (Pb) contamination in water, soil, and food crops. Six vegetables (Daucus carota, Brassica oleracea, Pisum sativum, Solanum tuberosum, Raphanus sativus, and Spinacia oleracea) were grown in the field under twelve treatments with different nutrient and water inputs. The lead concentrations in soil, vegetables for all treatments and water samples ranged from 1.038-10.478, 0.09346-9.0639 mg/kg and 0.036-0.26448 mg/L, The concentration of lead in soil treated with wastewater in treatment (T6) and vegetable samples was significantly higher, exceeding the WHO's permitted limit. Mineral and organic fertilizers combined with wastewater treatment reduced lead (Pb) concentrations in vegetables compared to wastewater application without organic fertilizers. Health risk indexes for all treatments except wastewater treatment (T 6 ) were less than one. Pb concentrations in mineral fertilizers, cow manure, poultry manure, leaf litter, and sugarcane bagasse treated were determined to pose no possible risk to consumers., (© 2024. The Author(s).)

Key Message: Thchit42 constitutive expression for fungal resistance showed synchronisation with leaf augmentation and transcriptome analysis revealed the Longifolia and Zinc finger RICESLEEPER gene is responsible for plant growth and development. Pelargonium graveolens essential oil possesses significant attributes, known for perfumery and aromatherapy. However, optimal yield and propagation are predominantly hindered by biotic stress. All biotechnological approaches have yet to prove effective in addressing fungal resistance. The current study developed transgenic geranium bridging molecular mechanism of fungal resistance and plant growth by introducing cassette 35S::Thchit42. Furthermore, 120 independently putative transformed explants were regenerated on kanamycin fortified medium. Primarily transgenic lines were demonstrated peak pathogenicity and antifungal activity against formidable Colletotrichum gloeosporioides and Fusarium oxysporum. Additionally, phenotypic analysis revealed ~ 2fold increase in leaf size and ~ 2.1fold enhanced oil content. To elucidate the molecular mechanisms for genotypic cause, de novo transcriptional profiles were analyzed to indicate that the auxin-regulated longifolia gene is accountable for augmentation in leaf size, and zinc finger (ZF) RICESLEEPER attributes growth upregulation. Collectively, data provides valuable insights into unravelling the mechanism of Thchit42-mediated crosstalk between morphological and chemical alteration in transgenic plants. This knowledge might create novel opportunities to cultivate fungal-resistant geranium throughout all seasons to fulfil demand., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Automobile exhaust releases different types of pollutants that are at great risk to the air quality of the environment and incidental distress to the nature of roadside plants. Mimusops elengi L. is an evergreen medicinal tree cultivated along the roadside of Lahore City. This research aimed to investigate physiological, morphological and genomorphic characteristics of M. elengi under the influence of air pollution from vehicles. Healthy and mature leaves were collected from trees on Canal Bank and Mall roads of Lahore as the experimental sites and control sites were 20 km away from the experimental site. Different physiochemical, morphological, air pollution tolerance index (APTI) and molecular analysis for the detection of DNA damage were performed through comet assay. The results demonstrated the mean accumulated Cd, Pb, Cu and Ni heavy metal contents on the leaves were higher than the control plants (1.27, 3.22, 1.32 and 1.46 μg mg -1 ). APTI of trees was 9.04. Trees in these roads significantly (p < 0.01) had a lower leaf area, petiole length and leaf dry matter content in comparison to control site. Increased comet tail showed that DNA damage was higher for roadside trees than trees in the control area. For tolerance of air pollution, it necessary to check the APTI value for the M. elengi at the polluted road side of Lahore city. For long-term screening, the source and type of pollutants and consistent monitoring of various responses given by the trees should be known., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Monolepta is one of the diverse genera in the subfamily Galerucinae, including 708 species and 6 sub-species worldwide. To explore the information on the mitogenome characteristics and phylogeny of the section "Monoleptites", especially the genus Monolepta , we obtained the newly completed mitochondrial genomes (mitogenomes) of four Monolepta species using high-throughput sequencing technology. The lengths of these four new mitochondrial genomes are 16,672 bp, 16,965 bp, 16,012 bp, and 15,866 bp in size, respectively. All four mitochondrial genomes include 22 transfer RNA genes ( tRNAs ), 13 protein-coding genes ( PCGs ), 2 ribosomal RNA genes ( rRNAs ), and one control region, which is consistent with other Coleoptera. The results of the nonsynonymous with synonymous substitution rates showed that ND6 had the highest evolution rate, while COI displayed the lowest evolution rate. The substitution saturation of three datasets (13 PCGs &#95;codon1, 13 PCGs &#95;codon2, 13 PCGs &#95;codon3) showed that there was no saturation across all datasets. Phylogenetic analyses based on three datasets ( ND1 , 15 genes of mitogenomes, and 13 PCGs &#95;AA) were carried out using maximum likelihood (ML) and Bayesian inference (BI) methods. The results showed that mitogenomes had a greater capacity to resolve the main clades than the ND1 gene at the suprageneric and species levels. The section "Monoleptites" was proven to be a monophyletic group, while Monolepta was a non-monophyletic group. Based on ND1 data, the newly sequenced species whose antennal segment 2 was shorter than 3 were split into several clades, while, based on the mitogenomic dataset, the four newly sequenced species had close relationships with Paleosepharia . The species whose antennal segment 2 was as long as 3 were split into two clades, which indicated that the characteristic of "antennal segment 2 as long as 3" of the true " Monolepta " evolved multiple times in several subgroups. Therefore, to explore the relationships among the true Monolepta , the most important thing is to perform a thorough revision of Monolepta and related genera in the future.

The present study aimed to determine the accumulation levels of arsenic in pasture soil, forage and animals. An atomic absorption spectrophotometer was used to determine the concentration of metals in the samples of soil, forage and blood plasma of horses. The level of arsenic ranged between 4.26 mg/kg (summer) and 4.66 mg/kg (winter) in soil samples and 2.67 mg/kg (summer) and 2.94 mg/kg (winter) in forage samples. In blood plasma samples, the mean arsenic (As) values varied between 1.38 and 1.52 mg/L. In the blood plasma samples, the mean As values varied between 1.38 and 1.52 mg/L. No statistically significant changes were observed for arsenic concentrations in plasma samples of horses for sampling seasons (p > 0.05). A positive correlation was observed for forage and blood plasma to a certain degree for arsenic but for other media, arsenic correlations were negative and insignificant. It is therefore suggested that regular monitoring of heavy metals in soils/plants/animals is essential to prevent excessive build-up of arsenic., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The primary objective of this study was to determine the heavy metal contents in the water-soil-coriander samples in an industrial wastewater irrigated area and to assess the health risks of these metals to consumers. Sampling was done from areas adjoining the Chistian sugar mill district Sargodha and two separate sites irrigated with groundwater (Site 1), and sugar mill effluents (Site 2) were checked for possible metal contamination. The water-soil-coriander continuum was tested for the presence of cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), manganese (Ni), lead (Pb), and zinc (Zn). The mean concentrations of all metals were higher than the permissible limits for all studied metals except for Mn in the sugar mill wastewater, with Fe (8.861 mg/L) and Zn (9.761 mg/L) exhibiting the highest values. The mean levels of Fe (4.023 mg/kg), Cd (2.101 mg/kg), Cr (2.135 mg/kg), Cu (2.180 mg/kg), and Ni (1.523 mg/kg) were high in the soil at Site 2 in comparison to the groundwater irrigated site where Fe (3.232 mg/kg) and Cd (1.845 mg/kg) manifested high elemental levels. For coriander specimens, only Cd had a higher mean level in both the groundwater (1.245 mg/kg) and the sugar mill wastewater (1.245 mg/kg) irrigated sites. An estimation of the pollution indices yielded a high risk from Cd (health risk index (HRI): 173.2), Zn (HRI: 7.012), Mn (HRI: 6.276), Fe (HRI: 1.709), Cu (HRI: 1.282), and Ni (HRI: 1.009), as all values are above 1.0 indicating a hazard to human health from consuming coriander irrigated with wastewater. Regular monitoring of vegetables irrigated with wastewater is strongly advised to reduce health hazards to people.

This study aimed to synthesize chitosan/polyvinyl alcohol (CS/PVA)-based zinc oxide (ZnO) and titanium dioxide (TiO 2 ) hybrid bionanocomposites (BNCs) and observe their comparative accomplishment against the skin cancer cell line, A431, and antioxidant potential. CS was blended with PVA to form polymeric films reinforced with the immobilization of ZnO and TiO 2 nanoparticles (NPs), separately. The optimization of the BNCs was done via physicochemical studies, viz. moisture content, swelling ratio, and contact angle measurements. The free radical scavenging activity was observed for 1,1-diphenyl-2-picryl-hydrazyl, and the antibacterial assay against the Escherichia coli strain showed a higher zone of inhibition. Furthermore, the anticancer activity of the synthesized BNCs was revealed against the skin cancer cell line A431 under varying concentrations of 50, 100, 150, 200, and 300 μg/mL. The anticancer study revealed a high percent of cancerous cell inhibition (70%) in ZnO BNCs as compared to (61%) TiO 2 BNCs in a dose-dependent manner.

Poultry feed comprises cereals and their by-products and is vulnerable to aflatoxins contamination. This study utilised reduced graphene oxide-titanium dioxide (rGO-TiO 2 ) nanomaterial as a dispersive solid phase extraction (d-SPE) adsorbent to extract, enrich and purify aflatoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2). The synthesis of rGO-TiO 2 nanomaterials through hydrothermal process and characterisation by transmission electron microscopy, scanning electron microscopy, Brunauer-Emmett-Teller (BET) and X-ray diffraction reveals that the nanomaterials have a single-layer structure embedded with TiO 2 nanoparticles. The matrix-spiked technique was employed for the extraction process, optimisation of d-SPE, and analytical method validation. The most appropriate extraction solvent was acetonitrile/water/formic acid (79/20/1, v/v/v), with 30 min of extraction time assisted by ultra-sonication. The optimised d-SPE parameters were: 50 mg of rGO-TiO 2 as sorbent amount, 2% methanol as the sample loading solvent, 30 min as adsorption time, and absolute ethanol as the washing reagent. The d-SPE method exhibited good desorption efficiency with 3 mL of acetonitrile/formic acid (99/1, v/v) and 20 min desorption time. After validation, the UHPLC-MS/MS analytical method has an acceptable range of specificity, linearity ( R 2  ≥ 0.999), sensitivity (LOQ 0.04-0.1 µg kg -1 ), recoveries (74-105% at three matrix-spiked levels) and precision (RSD 1.5-9.6%). Poultry feed samples ( n  = 12) were pretreated by this method to extract, enrich and analyse aflatoxins, which were detected in all poultry feed samples. The contamination levels were within the permissible limits.

Faisalabad is a major industrial area in Pakistan's Punjab province that discharges wastewater into the Chenab River. Industrial effluents in Faisalabad are predicted to pose a significant threat to the riparian vegetation of the Chenab River and nearby vegetation. Heavy metal pollution of plants, water, and soils is one of the biggest problems worldwide that needs to be addressed because heavy metals above normal levels are extremely dangerous to both riparian vegetation and wildlife. The results indicated high levels of pollution in the industrial effluents as well as in the river in terms of salinity, metal toxicity, TSS, TDS, SAR, the acidic and alkaline nature of the industrial effluents, and the spread of industrial effluents up to 15 square kilometres in the Chenab River. Despite the higher pollution, four plants were found at all sites: Calotropis procera , Phyla nodiflora , Eclipta alba and Ranunculus sceleratus . It was found that most of the selected plants were phytoaccumulators, making them best suited to survive in harsh environments such as those with industrial pollution. The Fe concentration in the plant constituents was the highest, along with Zn, Pb, Cd, and Cu, all of which were above the permissible limits of the WHO. The metal transfer factor (MTF) was higher in most of the plants studied, and even exceeded 10 at some severely affected sites. Calotropis procera proved to be the most suitable plant for growth on drainage systems and also at river sites, as it had the highest importance value across all sites and seasons., Competing Interests: All authors declare that they do not have any competing interests., (©2023 Abbas et al.)

Mucuna pruriens Linn. ( M. pruriens ), a leguminous plant, was used extensively in Ayurveda, to treat male-related infertility. Previous studies have demonstrated antioxidant, androgenic, aphrodisiac, and spermatogenic properties of M. pruriens seed extract. Surprisingly, the biological activities of M. pruriens on aging-induced pathological changes in the testis microenvironment have never been explored and the present study was focused on the testing therapeutic efficacy of M. pruriens on aged rat testis. Male Wistar albino rats were grouped as; adult (3 months), aged (24 months), aged +  M. pruriens and adult +  M. pruriens ( N  = 6/group). The extract was administrated at a dose of 200 mg/kg body weight (dosage determined in our previous study) daily by gavage for 60 days. The total and free testosterone, FSH and LH levels were considerably increased in aged +  M. pruriens . The diameter & volume of the seminiferous tubules, the height & volume of the epithelium, and the number of Leydig cells number were significantly decreased in aged rat testis, concomitantly connective tissue proportion was increased compared to adult rats. The seminiferous epithelium indicates significant rejuvenation or restoration of spermatogenic cells in aged +  M. pruriens rat testis. The highlighting observations in aged +  M. pruriens was increased in the following parameters i.e., tubular diameter (25%), number of tubules (35%), epithelial height (25%) & volume (20%), and number of Leydig cells (35%) when compared to untreated aged rat testis. The TNFα, NF-κB, cytochrome c, Caspase-9, Caspase-3, Bcl-2, Bax, PARP iNOS, and inflammatory and apoptotic factors were downregulated in aged +  M. pruriens . M. pruriens was able to restore spermatogenesis and enhance the activity of Sertoli cells and Leydig cells and improve the pituitary-gonadal axis in aged rat testis and observations indicate the therapeutic activity of M. pruriens in aged rat testis., Competing Interests: Conflict of interestNone of the authors has any financial/commercial conflicts of interest to declare., (© King Abdulaziz City for Science and Technology 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Cobalt (Co) bioaccumulation, contamination, and toxicity in the soil environment, plant growth, and cattles' health are becoming a severe matter that can cause unembellished consequences in environmental safety and human health. The present research was conducted for the assurance of cobalt (Co) amassing in three forage plant species (Zea mays, Sorghum bicolor, Trifolium alaxandrium), from four ecological sites, and sewage water and in buffaloes blood was investigated. The analysis of variance showed significant differences for Co concentration in the soil and sewage water collected from all ecological sites. Meanwhile, summer and winter seasons and forage ecotypes significantly influenced the quantity of Co. The forage pastures also vary significantly in the concentration of Co in the above-ground parts. The highest Co level was present in Trifolium alaxandrium at ecological site-5. Cobalt taken from wastewater had a higher concentration in Trifolium alaxandrium during the winter. The samples which are collected from site-V and site-IV have the maximum concentration of Co because these areas receive highly contaminated water for irrigation. Cobalt tends to be bioaccumulated in the food chain and can cause serious problems in humans and animals. Bioaccumulation of cobalt in collected samples could be accredited to anthropogenic activities. Pollution load index values for all samples fell in the range below 1. The health risk index indicated the probability of health damage caused by the ingestion of contaminated fodder. An increase of Co concentration in soil, fodder, and blood owing to wastewater irrigation to crops was indicated as an outcome of this investigation. The results indicate that the Co toxicity in forage crops is attributed to Co bioaccumulation, transfer, and pollution load in the soil-water-cattle triangle. Efforts should be extended to avoid contamination of the food chain via Co-rich sewage water. Other nonconventional water resources should be used for forage irrigation., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

With the emergence of delay- and energy-critical vehicular applications, forwarding sense-actuate data from vehicles to the cloud became practically infeasible. Therefore, a new computational model called Vehicular Fog Computing (VFC) was proposed. It offloads the computation workload from passenger devices (PDs) to transportation infrastructures such as roadside units (RSUs) and base stations (BSs), called static fog nodes. It can also exploit the underutilized computation resources of nearby vehicles that can act as vehicular fog nodes (VFNs) and provide delay- and energy-aware computing services. However, the capacity planning and dimensioning of VFC, which come under a class of facility location problems (FLPs), is a challenging issue. The complexity arises from the spatio-temporal dynamics of vehicular traffic, varying resource demand from PD applications, and the mobility of VFNs. This paper proposes a multi-objective optimization model to investigate the facility location in VFC networks. The solutions to this model generate optimal VFC topologies pertaining to an optimized trade-off (Pareto front) between the service delay and energy consumption. Thus, to solve this model, we propose a hybrid Evolutionary Multi-Objective (EMO) algorithm called S warm O ptimized N on-dominated sorting G enetic algorithm (SONG). It combines the convergence and search efficiency of two popular EMO algorithms: the Non-dominated Sorting Genetic Algorithm (NSGA-II) and Speed-constrained Particle Swarm Optimization (SMPSO). First, we solve an example problem using the SONG algorithm to illustrate the delay-energy solution frontiers and plotted the corresponding layout topology. Subsequently, we evaluate the evolutionary performance of the SONG algorithm on real-world vehicular traces against three quality indicators: Hyper-Volume (HV), Inverted Generational Distance (IGD) and CPU delay gap. The empirical results show that SONG exhibits improved solution quality over the NSGA-II and SMPSO algorithms and hence can be utilized as a potential tool by the service providers for the planning and design of VFC networks.

Irrigation using sewage water can be beneficial, as it can increase the productivity of crops but has negative consequences on crops, soil contamination, and human health. It contains a variety of toxins, such as chemicals and heavy metals, which damage the soil and crops. In this regard, the aim of the research was to assess the potential health hazards of iron (Fe) metal in food crops (leafy and root crops) treated with wastewater (T&#95;1), canal water (T&#95;2), and tube well water (T&#95;3). Water, soil, and edible components of food crops were collected at random from three distinct locations. Fe concentration in samples was estimated using atomic absorption spectrophotometer, following wet digestion method. The Fe concentrations, ranged from 0.408 to 1.03 mg/l in water, 31.55 to 187.47 mgkg-1 in soil and 4.09 to 32.583 mgkg-1 in crop samples; which were within permissible limits of the World Health Organization (WHO). There was a positive correlation between soils and crops. The bioconcentration factor, enrichment factor (EF), daily intake of metals (DIM), health risk index (HRI), and target hazard quotient (THQ) all values were &lt;1, except for a pollution load index &gt;1, which indicated soil contamination, but there was no Fe toxicity in crops, no health risk, and no-carcinogenic risk for these food crops in humans. To prevent the excessive accumulation of Fe metal in the food chain, regular monitoring is needed., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Akhtar et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Birth preparedness and complication readiness (BPCR) is a broad system to increase the practice of trained health provision at the time of childbirth and the key interventions to decrease mothers' and newborns' death. However, its status and influencing factors have not been well studied at different levels in the study area. The current study aimed to assess the BPCR status and explore its associated factors influencing BPCR among childbearing age women in Thatta, District of Sindh., Methods: This community-based cross-sectional study was conducted among 770 recently delivered mothers from October 2016 -September 2017, recruited using a multistage cluster sampling technique. A structured validated close-ended questionnaire measuring BPCR knowledge and practices was used for the interviews. The results were analyzed by means of the Chi-square test, and a binary logistic regression model was used to determine the factors influencing BPCR., Results: The overall response rate was 94.6%, with a low BPCR status. Out of 770 participants, only 163 (21.2%) were well prepared, while 607 (78.8%) were not prepared for safe childbirth and its complications. A small proportion of women knew about the serious warning signs of pregnancy, labour, childbirth and the postpartum period (16.2%), (15.3%) and (22.7%) respectively. Antenatal care (ANC) checkup (P < 0.001), cost of ANC checkup (p = 0.016), place of birth (p = 0.014), awareness of serious warning signs during pregnancy (p = 0.001) and awareness of serious warning signs during the postpartum period (p < 0.001) were found to be significant predictors of BPCR., Conclusion: The proportion of women who were well prepared for birth and its complications was low. It is recommended to organize community-based education campaigns and improve the quality of MNCH services at every level to increase BPCR among women in Sindh., Competing Interests: The authors have declared that no competing interests exist.

Purpose: PARK2 is a potential tumour suppressor gene and its genetic alterations (regionic loss) are common across many human cancers. The association of PARK2 germline variations (SNPs) with Parkinson's has been shown, but their association in development and progression of cancer remains elusive. The aim of this study was to identify association of PARK2 polymorphisms (rs1801474, rs1801334) with colorectal cancer in a case control study design., Methods: This case control study included a total of 650 genetically unrelated subjects comprising 300 colorectal cancer cases and 350 healthy controls belonging to North Indian. Both SNPs were analyzed using the PCR-RFLP assay. Statistical analysis for describing risk and association was performed using SPSS-17 software. Structural deviations due to non- synonymous substitutions (S167N and D394N) were analyzed using MD simulations., Results: The genotype distributions of both the SNPs were in Hardy-Weinberg equilibrium. For both the polymorphisms, the allelic model showed statistically significant risk with OR ~ 1.3. Many of the associations remained significant even after Bonferroni correction (P < 0.00125). The result suggested that both S167N and D394N were deviated from wild type and structures and were stable after 5 ns. The average value of RMSD for backbone atoms was calculated from 5 to 10 ns molecular dynamics simulation data., Conclusion: In conclusion, our study revealed a significant association of PARK2 SNPs with colorectal cancer as well as their relations with other clinical parameters highlighting their contribution towards colorectal cancer susceptibility in North Indian population., (© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)

Environmental extremes such as hypersaline conditions are significant threats to agricultural productivity. The sustainable use of halophilic microbial strains was evaluated in plant in a salt stress environment. Oxygen-evolving complex (OEC), energy compartmentalization, harvesting efficiencies (LHE), specific energy fluxes (SEF), and nitrogen assimilation of oilseed crops (Sunflower cultivars) in a suboptimal environment was examined. Plants were grown in a plastic pot (15 ×18 cm 2 ) containing sterilized (autoclaved at 120°C for 1 h) soil. Twenty-five ml suspension (10 7 CFU/ml) each of Bacillus cereus strain KUB-15 and KUB-27 (accession number NR 074540.1) and Bacillus licheniformis strain AAB9 (accession number MW362506), were applied via drenching method. Month-old plants were subjected to salt stress via gradual increment method. The energy compartmentalization of microbial inoculated plants exposed to salt stress revealed higher photosystem II (PSII) activity at the donor side, lesser photo-inhibition, and increased performance of oxygen-evolving complex compared to control. High potassium (K + ) and low sodium (Na + ) ions in treated leaves with the activated barricade of the antioxidant system stimulated by Bacillus strains favored enhanced photochemical efficiency, smooth electron transport, and lesser energy dissipation in the stressed plants. Moreover, the results reveal the increased activity of nitrite reductase (NiR) and nitrate reductase (NR) by microbial inoculation that elevated the nitrogen availability in the salt-stressed plant. The current research concludes that the application of bio-inoculants that reside in the hyper-saline environment offers substantial potential to enhance salt tolerance in sunflowers by modulating their water uptake, chlorophyll, nitrogen metabolism, and better photochemical yield., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ali, Wei, Siddiqui, Chen, Ansari, Wajid, Shams, Abbasi and Zafar.)

Salinity is a major threat to agricultural productivity worldwide. The selection and evaluation of crop varieties that can tolerate salt stress are the main components for the rehabilitation of salt-degraded marginal soils. A field experiment was conducted to evaluate salinity tolerance potential, growth performance, carbon (δ 13 C) and nitrogen isotope composition (δ 15 N), intrinsic water use efficiency (iWUE), harvest index, and yield stability attributes in six barley genotypes (113/1B, 59/3A, N1-10, N1-29, Barjouj, Alanda01) at three salinity levels (0, 7, and 14 dS m -1 ). The number of spikes m -2 was highest in Alanda01 (620.8) while the lowest (556.2) was exhibited by Barjouj. Alanda01 produced the highest grain yield (3.96 t ha -1 ), while the lowest yield was obtained in 59/3A (2.31 t ha -1 ). Genotypes 113/1B, Barjouj, and Alanda01 demonstrate the highest negative δ 13 C values (-27.10‱, -26.49‱, -26.45‱), while the lowest values were obtained in N1-29 (-21.63‱) under salt stress. The δ 15 N was increased (4.93‱ and 4.59‱) after 7 and 14 dS m -1 as compared to control (3.12‱). The iWUE was higher in N1-29 (144.5) and N1-10 (131.8), while lowest in Barjouj (81.4). Grain protein contents were higher in 113/1B and Barjouj than other genotypes. We concluded that salt tolerant barley genotypes can be cultivated in saline marginal soils for food and nutrition security and can help in the rehabilitation of marginal lands.

Background: The flux of isoprenoids and the total accumulation of triterpenoid saponins known as centellosides in C. asiatica are controlled by the key genes of the Mevalonate pathway (MVA). These genes were reported to have positive regulation of the pathway in providing isoprenoid moieties. Though, some information is available on the pathway and secondary metabolites. However, most of the pathway steps are not characterized functionally., Methodology and Results: For the study, full-length pathway gene Hydroxymethyl glutaryl-CoA-synthase (CaHMGS; GenBank accession number: MZ997833), was isolated from previously annotated transcriptome data of Centella asiatica leaves. HMGS has been successfully cloned and heterologously expressed in bacteria E. coli strain DH5α. The cloned gene has been sequenced and further characterized through in silico studies by different bioinformatics tools. Also, the gene sequences have been submitted in NCBI. In silico studies of isolated gene sequence revealed the nature, characteristics of genes. The ORF of HMGS is 1449 bp encoding 482 amino acids. Predicted molecular weight (MW) of HMGS was 48.09 kDa and theoretical pI was 5.97. Blast results and Multiple sequence alignments of the gene showing the similarity with HMGS of other plants of their respective families. The Molecular Evolutionary Genetic Analysis (MEGA) version 10.1.6 was used to construct a phylogenetic tree. Differential tissue-specific expression of different plant parts was also checked. Tissue expression patterns unveiled that the highest expression level of the CaHMGS had been seen in the roots and lowest in the node of the plant. Functional complementation experiment of the CaHMGS in Saccharomyces cerevisiae wild strain YSC1021 and haploid strain YSC1021 which lack HMGS protein confirmed that the CaHMGS gene encodes functional CaHMGS that catalyzed the biosynthesis of mevalonate in yeast., Conclusions: The gene was reported, cloned and characterized first time in Centella asiatica. Understanding this biosynthetic pathway gene will further help in the improvement of plants for enhanced secondary metabolites production., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Pollution caused by municipal solid waste (MSW) is becoming a serious threat to the environment. Composting may be an effective way to speed up the decomposition of biodegradable components in MSW, resulting in compost that can be utilized as an organic fertilizer. The pot experiments were carried out with different soil-MSW mixtures (100:0, 75:25, 50:50, and 25:75; w/w) to determine the impact of MSW on the bioconcentration of Cd in commonly consumed plants of Sargodha. The possible health risks were evaluated by applying pollution indices, such as the pollution load index, bioconcentration factor, enrichment factor, and health risk index. The pollution load index was higher than 1 in 75% MSW-amended soil. However, the concentration of Cd was found to be below the permissible limits in all studied vegetables, with a range of 0.019-0.106 mg/kg. In the study, serum samples from different volunteers living in four sites in Sargodha were also collected and analyzed. For vegetable crops, the health risk index (HRI) was less than one. It is concluded that the concentration of Cd was increased by increasing the fraction of MSW. Although the metal contents in the soil treated with MSW were not high enough to categorize the soil as polluted, these findings show that the reuse of MSW can serve as an alternative to mineral fertilizers. However, the presence of Cd in MSW can have a direct impact on soil fertility and, if biomagnified, on crop production and human health., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

In South Asia, the high costs of synthetic fertilizers have imposed research on alternative nutrient inputs. We aimed to identify potential trace elements (PTE) present in some organic manure that might be a source of environmental pollutions and risk to public health following consumption. The study aims to evaluate how different organic waste (poultry waste, PW; press mud, PM; and farmyard manure, FYM) influences the heavy metal migration in soil, the accumulation in garlic, and their potential health risks. Organic waste caused a higher accumulation of certain metals (Zn, Cu, Fe, and Co), whereas Mn, Cd, Cr, and Pb were in lower concentrations in soil. Amendments of soil with PM revealed a higher accumulation of Cd, Cr, Fe, and Pb, whereas PW resulted in Cu and Zn accumulation in garlic. Treatment of soil with FYM exhibited higher metal concentration of Co and Mn. An environmental hazard indices study revealed that pollution load index (PLI) was highest for Cu following treatment with PM. The health risk index (HRI) was greater for Cd following amendment with PM. Maximum daily intake of metals (DIM) was observed for Zn after treatment with PW. The Pb exhibited maximum bioconcentration factor (BCF) in PM-amended plants. Based on these findings, we concluded that garlic grown on contaminated soil with organic waste may pose serious health hazards following consumption.

Present study was conducted to check the heavy metal content in wheat treated with municipal solid waste, Although municipal solid waste was enriched with organic nutrient, micronutrients, and macro-nutrients, considerable amount of cobalt was also witnessed in municipal solid waste that s why pot experiment was executed. The concentration of cobalt in different parts of wheat (root, shoot, and grain) was analyzed by atomic absorption spectrophotometer (AA-6300 Shimadzu Japan). Highest concentration of cobalt in root, shoot, and grain of wheat was observed in the range of 0.91-1.02 mg/kg, 0.92-1.04 mg/kg, and 0.93-1.00 mg/kg, respectively, under the influence of different fertilizer's used, while in field experiment, level of Co was greater in roots followed by shoots and grain. The metal concentration in wheat grown in field was ranged from 0.67 to 0.72 mg/kg for roots, 0.64 to 0.71 mg/kg for shoots, and 0.66 to 1.71 mg/kg for grains. Concentrations of cobalt were found below the permissible limits suggested by FAO/WHO. Various indices (PLI, BCF, DIM, HRI) were calculated and results showed that PLI was above 1 indicating that metal was causing pollution in treatments while value of BCF, DIM, and HRI was within the permissible range. Higher Co content in wheat may cause damage to the pancreatic cells of animals, cause respiratory problems, and effects their kidney, liver, and lungs, if exposure is for long period through feed., (© 2022. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Zinc (Zn) is a vital nutrient element required for plants normal growth and development. It performs imperative functions in numerous metabolic pathways in the plants. However, potentially noxious levels of Zn in terrestrial environment can lead to inhibited photosynthesis, growth, respiratory rate and imbalanced mineral nutrition. In micronutrient malnutrition, Zn deficiency is a global human health problem owing to the human dependence on cereals grains especially wheat-based diet. Therefore, this study investigated the Zn uptake efficacy in Triticum aestivum that is grown under two different doses (100 g/kg or 200 g/kg) of various soil amendments in both pot and field experimentation. Results of this study revealed that mean Zn concentration in different wheat varieties and treatments were varied from 1.53 to 6.03 mg/kg, 11.27 to 40.65 mg/kg, 11.28 to 39.93 mg/kg, and 11.32 to 37.70 mg/kg in amended soil, root, shoot, and grains, respectively. All observed Zn values in soil and wheat parts were lower than the FAO/WHO standards. Zinc values observed for pollution load index (0.034-0.134 mg/kg), daily intake (0.00492-0.01533 mg/kg), and health risk (0.0164-0.0570 mg/kg) index were lower than 1 except bio-concentration factor. Bio-concentration factor (5.076-10.165 mg/kg) revealed that DHARABI-11 variety showed maximum Zn uptake efficacy in farmyard manure treatment. The daily intake and health risk index values also showed that Zn level in grains is safe for inhabitants consumption. Overall, study recommended that these organic amendments are a good source of fertilizers, essentially required for the sustainable management of soil and increases the Zn accumulation in wheat grains which can ultimately reduce the Zn malnutrition in human food chain., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

In District Jhang, farmers use municipal wastewater to irrigate fodder crops as an alternative source to the deficient availability of fresh water. Therefore, the present study selected the three irrigation sources in District Jhang (canal water, ground water and municipal wastewater) to study the iron (Fe) concentration in the soil, fodder crops and ultimately their transfer into the animal body. Analysed Fe concentration varied as 16.40-27.53 mg/kg in soil samples, 19.72-30.34 mg/kg in fodder crops and 2.49-5.11 mg/kg in animals. Analysed Fe concentration in soil was higher on the wastewater irrigation site while canal water-irrigated fodder crop Zea mays exhibit the higher Fe concentration. In animal samples, higher Fe concentration was observed in the cow blood (4.09 mg/l), cow hairs (3.39 mg/kg) and cow faeces (5.11 mg/kg). Results of pollution load index (0.288-0.484 mg/kg) and enrichment factor (0.112-0.197 mg/kg) indicated that Fe concentration was minimally dispersed and enriched in these sites. Health risk and daily intake values were observed between the 0.029-0.059 and 0.042-0.084 mg/kg/day. Bio-concentration factor (0.834-1.47 mg/kg) for Fe which was greater than 1 explains that Fe contamination was transferred from the soil to fodder tissues and may raise health issues in the grazing animals if they are continuously exposed to these contaminated forages. Wastewater irrigation in study area has increased the Fe content in soil-plant environment that is a risking factor for animal and human health. Hence, this study recommended that wastewater should be treated prior to their irrigation on agricultural lands., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Many studies have described the physiological, biochemical, and molecular responses to heavy metal toxicity and deficiencies individually in plants. The present study assess nickel (Ni) concentration in amended soil, plant vegetative parts, and wheat grains, grown under diverse types of fertilizers in Sargodha, Punjab, Pakistan. Different varieties of wheat were grown in pot and fields. Different treatments (municipal solid waste, poultry waste, press mud, farm yard manure) of fertilizers were applied in order to study the metal level increased in different parts (root, shoot, grain) of wheat due to fertilization. Results indicated that metal level was found highest in roots followed by shoot and grain. The highest level of nickel in root was present in V1 (2.35 mg/kg) due to T2 (2.60 mg/kg) treatment. Higher nickel levels in wheat shoot and grains were observed in V5 (2.36 mg/kg) and V8 (2.29 mg/kg), respectively, due to applied treatment T2 (2.57 mg/kg). This study concluded that treatment T9 was proven safe in view of the observed Ni concentration, while treatment T2 (municipal solid waste) resulted in higher accumulation of nickel in wheat grains which showed that municipal solid waste should be treated before their application in agriculture fields to secure the public health. This study recommended that although application of fertilizers increased the plant growth and nutritional value, it also enhanced metal accumulation in the wheat grains which could be harmful for consumers especially human being. Government should take actions to prevent metal toxicity in human food chain., (© 2021. Crown.)

Objectives: This study aimed to assess the time to sputum culture conversion (SCC) and its determinants among multidrug-resistant tuberculosis (MDR-TB) patients., Methods: This cross-sectional study was conducted from January 2019 to January 2020. A total of 252 MDR-TB patients presenting at a tertiary level teaching hospital in Peshawar, Khyber Pakhtunkhwa (KP), were included. The patient's demographic and clinical data were collected using a structured questionnaire. Time to SCC was calculated from the initiation of treatment till the patient had two consecutive negative cultures. The Cox proportional-hazards analysis was performed to check strength and association between the determinants and time for SCC., Results: Out of 252 MDR-TB patients enrolled, sputum culture conversion was observed in 76.6% of the patients by the end of six months. While, 19.0% of the patients failed to achieve negative culture and remained positive after interim report of their treatment. Age > 45 years (HR = 15.22; 95% CI: 7.27-31.83; p<0.001), female gender (6.22; 2.90-13.36; p<0.001), BMI < 18.5 kg/m 2 (10.28; 5.25-20.11; p<0.001), weight loss (0.03; 0.01-0.06; p<0.001), smoking (0.10; 0.05-0.21; p<0.001), diabetes mellitus (0.02; 0.00-0.04 p<0.001) and disease severity on chest X-ray (CXR) (0.03; 0.01-0.09; p<0.001) were the significant determinants of delayed sputum culture conversion., Conclusion: MDR-TB patients with older age, low BMI, weight loss, diabetes, smokers and those with disease severity on CXR are less likely to respond to treatment as they displayed delayed SCC. Therefore, such patients should be meticulously followed up for successful management., Competing Interests: Conflicts of Interest: The author(s) declare no conflicts of interest., (Copyright: © Pakistan Journal of Medical Sciences.)

Heavy metal pollution in soil, forage, and animals is serious concern nowadays. Current research was conducted in Sargodha to find out the relationship of animals related to the forages and soil pollution. Three sites were selected with three different treatments; site I irrigated with ground water, site II irrigated with the canal water, and site III irrigated with the wastewater. Samples of soil, forage, and animals (blood, hair, feces) were collected from selected sites and were analyzed for metal analysis using atomic absorption spectroscopy. Results indicated that Zn in soil ranged from 24.12 to 37.39 mg/kg; forage, 31.98-44.47 mg/kg; blood of animals, 1.49-2.72 mg/L; hair of animals, 1.37-2.41 mg/kg; and feces of animals, 1.06-2.97 mg/kg. The concentration of zinc in soil and forage was less than permissible limit, but concentration in blood of animals was greater than critical limit suggesting the presence of metal. Bio-concentration factor indicated that metal was accumulated in forages growing at irrigated site. HRI concentration (2.024 mg/kg/day) suggests the accumulation of zinc in animal tissues. Pollution load index and enrichment factor were within the range., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

This study's aim was to investigate iron (Fe) and zinc (Zn) concentration in the soil, forage crops, and sheep blood with respect to the seasonal availability of these metals. Soil, forage, and sheep blood samples were sampled from five different locations in Chakwal (Pidh, Tobar, Ratoccha, Choa Saiden Shah-Kalar Kahar road, and Choa Saiden Shan-Chakwal Road) during two seasons, i.e., winter and summer. All the samples were processed through wet acid digestion for evaluation of metal contents. Because of proximity of site-1 and site-2 to coal mines, higher Fe concentration was observed than Zn. Overall, varied Fe concentrations obtained in soil were 12.95-24.31 mg/kg, 1.29-9.61 mg/kg in forage and 1.17-24 mg/l in blood, whereas Zn values were 1.04-31.9 mg/kg, 1.96-7.02 mg/kg, and 0.16-6.52 mg/l for soil, forages, and blood respectively. The pollution load index value for both Fe (0.01-0.14 mg/kg) and Zn (0.02-0.72 mg/kg) was lesser than 1. Bio-concentration (0.09-2.64mg/kg) and enrichment factor (0.08-7.51 mg/kg) were showing efficient transfer of metals through the food chain. Daily intake and health risk index values of iron were ranged from 0.01 to 1.1 mg/kg/day and 0.02 to 1.05 mg/kg/day. There was a probable chance of upsurge in metal values in coming years due to continued mining activities. Anthropogenic input, mainly mining activities in the study area, have increased the Fe and Zn content in the environment which can ultimately find their way up the food chain, thereby risking the health of grazing livestock., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Heavy metals pollution has thorough worldwide apprehensions due to the instantaneous growth of industries. Farming regions are irrigated mainly with wastewater which contains both municipal and industrial emancipations. Keeping in view the above scenario, a study was designed in which three sites irrigated with ground, canal, and municipal wastewater in the District Jhang were selected to determine the zinc accumulation and its transfer in the soil, plant, and animal food chain. Zinc concentration was ranged as 18.85-35.59mg/kg in the soil, 26.42-42.67 mg/kg in the forage, and 0.982-2.85mg/kg in the animal samples. Investigated zinc concentration in soil and forages was found to be within the recommended WHO/FAO limits, but blood samples exceed the standards of NRC (2007). The maximum level of pollution load index (0.427-0.805mg/kg) and enrichment factor (0.373-0.894 mg/kg) for zinc was noticed upon wastewater irrigation. Daily intake (0.039 to 0.082 mg/kg/day) and health risk index (0.130 to 0.275 mg/kg/day) of zinc metal was higher in the buffaloes that feed on wastewater-irrigated forages. Bio-concentration factor (0.840 to 2.01mg/kg) for soil-forage was >1 which represents that these plants accumulated the zinc concentration into their tissues and raised health issues in grazing animals on consumption of wastewater-contaminated forages. As animal-derived products are part of human food, then zinc toxicity prevailed in livestock tissues ultimately affects the human food chain. Overall, findings of this study concluded that animal herds should be monitored periodically to devise preventive measures regarding the toxic level of heavy metals availability to livestock., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Grazing animals act as a bioindicator to study the heavy metal status in the pasture lands because excessive amount of toxic metals in the animal diet either disturb their normal activity or deposit the contaminants into their tissues. The aim of this study was to appraise the chromium status in soil and pasture crops with respect to the nutritional requirement of grazing animals. Three different sites were selected to collect soil, forages, and animal samples from District Jhang. All the samples were processed through atomic absorption spectrophotometer to analyze the chromium concentration in them. Chromium concentration was varied as 0.703-4.20 mg/kg in soil, 0.45-2.85 mg/kg in forages, and 0.588-2.37 mg/kg in all collected animal samples. Both the soil and forage samples displayed the maximum chromium concentration in the Capparis decidua, whereas animal samples revealed maximum concentration in animal blood. Results of pollution load index (0.078 to 0.463 mg/kg) exhibited that all the sample values are less than unity while enrichment factor (1.57-8.25mg/kg) showed that significant level of chromium is enriched in these sites. The maximum value of daily intake (0.0007-0.0055mg/kg/day) and health risk index (0.0004-0.00370055mg/kg/day) was observed in the buffalo that feed on the Capparis decidua. Bio-concentration factor (0.398-2.09mg/kg) value was the maximum in the Medicago sativa. It is concluded that all the animal samples showed chromium concentration beyond their standards. Thus, proper measures should be taken to reduce the metal contamination in these areas that ultimately lessen the availability of toxic metals to grazing animals., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

The United Nation's Sustainable Development Goals include the target of ensuring access to water and sanitation and hygiene (WASH) for all; however, very few studies have assessed comprehensive school WASH service in Pakistan. The purpose of this study was to identify WASH services in primary schools of Pakistan, and to assess how recent WASH interventions and policies are associated with the school's academic performance. A representative cross-sectional study was conducted in primary schools in the Sindh province of Pakistan. Structured observations and interviews were done to ascertain the schools' WASH conditions. The primary exposures of interest were the implementation of previous WASH interventions and National WASH policy in the school and the WASH coverage. Outcomes of interest included WASH conditions and school performance. The structural equation modeling (SEM) using a bootstrap resampling procedure was employed to characterize how WASH exposures were associated with WASH conditions and school performance. Data were collected from 425 schools. The Basic WASH facilities coverage in the primary schools of Sindh remains overall low according to WHO WASH service ladder criteria. Also, inconsistency in all three inclusive domains of WASH (availability, accessibility, and functionality) facilities were found. The school performance was significantly associated (P<0.001) with the presence of WASH interventions and/or WASH policy, while WASH policy and/or recent WASH intervention at the school were not associated with overall water quality. Our assessment unveiled several WASH gaps that exist, including high heavy metal and fecal contamination. Adoption of national WASH policy and financing of evidence-based WASH interventions are recommended in primary schools to improve educational outcomes., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Present work evaluated the zinc (Zn) concentration in soil, forage, blood plasma, hair, and feces samples of cows, buffaloes, and sheep taken from Mianwali, Punjab, Pakistan. The concentration of Zn was found in the ranged of 21.82-35.09mg/kg, 32.59-42.17mg/kg, 0.927-2.48mg/l, 1.03-2.84mg/kg, and 0.923-1.98mg/kg in soil, forage, blood plasma, hair, and feces samples, respectively. The Zn concentration in soil, forage, blood, hair, and feces was safer compared to standard limits. Statistical analysis described that values for BCF, PLI, EF, DIM, and HRI ranged 1.03-1.57mg/kg, 0.486-0.782mg/kg, 0.457-0.696mg/kg, 0.048-0.08mg/kg, and 0.160-0.272mg/kg, respectively. It can be concluded from the present work that Zn concentration was safe in soil, forages, and animal samples. BCF was noticed as greater than 1 while PLI, EF, DIM, and HRI were found less than 1, so regular heavy metal analysis was required to appraise the contamination level in environment., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Scabies, a human skin infestation caused by the ectoparasitic mite Sarcoptes scabiei var. hominis , affects more than 200 million people globally. The prevailing knowledge of the disease process and host immune response mechanisms is limited. A better understanding of the host-parasite relationship is essential for the identification of novel vaccine and drug targets. Here we aimed to interrogate the transcriptomic profiles of mite-infested human skin biopsies with clinical manifestations of ordinary scabies subjects ("OS"; n = 05) and subjects naive to scabies ("control"; n = 03) using RNASeq data analysis. A combined clustering, network, and pathway mapping approach enabled us to identify key signaling events in the host immune and pro-inflammatory responses to S. scabiei infestation. The clustering patterns showed various differentially expressed genes including inflammatory responses and innate immunity genes (DEFB4A, IL-19, CXCL8, CSF3, SERPINB4, S100A7A, HRNR) and notably upregulation of the JAK-STAT pathway in scabies-infested samples. Mite-infested human skin biopsies (GSE178563) were compared with an ex-vivo porcine infested model (E-MTAB-6433) and human skin equivalents (GSE48459). Marked enrichment of immune response pathways (JAK-STAT signaling, IL-4 and IL-13 pathway, and Toll receptor cascade), chemokine ligands and receptors (CCL17, CCL18, CCL3L1, CCL3L3, CCR7), and cytokines (IL-13 and IL-20) were observed. Additionally, genes known for their role in psoriasis and atopic dermatitis were upregulated, e.g., IL-19. The detailed transcriptomic profile has provided an insight into molecular functions, biological processes, and immunological responses and increased our understanding about transcriptomic regulation of scabies in human., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Shehwana, Ijaz, Fatima, Walton, Sheikh, Haider and Naz.)

Objective: To determine the reliability of knee joint sonography in the evaluation of gouty arthritis. Methodology: A search of Google Scholar, PubMed, NCBI, MEDLINE, and Medscape databases, from 1988 up to 2020. The key search terms used were knee joint; knee joint ultrasound; gout; gouty arthritis, knee joint pain; sensitivity; specificity. The reviewer independently screened the titles and abstracts of the relevant articles and full-text downloads to determine whether the inclusion or exclusion criteria were met. Results: In total, 103 articles were identified through the database search. In addition, 11 articles were identified through other sources. Then, screening was performed, and 9 articles were removed due to duplication. Further screening was done for 105 articles, and 27 articles were excluded due to insufficient information. Seventy-eight full-text articles were assessed for eligibility. A total of 13 full-text articles were excluded due to research performed on animals, as the study had been designed as a review of only human studies. Sixty-three studies were included that had a qualitative synthesis. Conclusion: The knee is a weight-bearing joint and may be affected by a myriad of different pathological conditions, therefore a proper diagnosis is of prime importance for a proper management plan. Ultrasound is a non-invasive, radiation-free, and readily available modality that has high sensitivity and specificity in the evaluation of gouty arthritis., Competing Interests: Conflict of interest The authors have no conflict of interest., (© Polish Ultrasound Society. Published by Medical Communications Sp. z o.o.)

The current study probed methicillin resistant S. aureus from milk of different dairy farms along with its response to multiple antibiotics, assessment of risk factors, and response to antibiotic coupled nanoparticle. XRD of Np was confirmed as miller indices (hkl) values i.e. (101), (100), (002), (110), (012) and (013) while STEM finally revealed 40-60 nm nanorods in aggregated form. Total of 6 preparations viz a viz gentamicin (G), chloramphenicol (C), zinc oxide nanoparticle (Np), gentamicin coupled Np (GNp), chloramphenicol coupled Np (CNp), and simultaneously coupling of gentamicin and chloramphenicol on Np (GCNp) were formulated for their potential to bring resistance modulation. Data analysis of this study revealed 24.59% MRSA from dairy milk appearing potentially associated (OR> 1, p < 0.05) with most of assumed risk factors. MRSA in response to various antibiotics showed highest resistance against amoxicillin (100%), penicillin (100%), vancomycin (100%), and linezolid (90%). Zone of inhibitions were increased by 249.76% (GNp), 184.86% (CNp), and 279.76% (GCNp) in case of coupled preparations. Significant reduced minimum inhibitory concentration was observed in case of GCNp (7.8125 ± 0.00 μg/mL) followed by GNp (15.00 ± 0.00 μg/mL) and CNp (41.67 ± 18.042 μg/mL) as compared to Np alone (125.00 ± 0.00 μg/mL). Minimum bactericidal concentrations for GCNp, GNp, and CNp, and Np were 31.125, 62.5, 125, and 500 μg/mL, respectively. The study thus concluded increased prevalence of MRSA while coupling of ZnO nanoparticles with antibiotics significantly brought resistance modulation to MRSA., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The aim of this study was to access the Mn contamination in soil, forages, and animals. Heavy metal pollution is a matter of prime significance in natural environment. Through food chain, toxicity of heavy metals and their bioaccumulation potential are transferred into humans. Higher concentrations of metallic compounds are toxic to living organisms but these are essential to maintain body metabolism. Intake of food crops polluted with heavy metals is chief food chain channel for human exposure. Animals are exposed to heavy metal stress by the intake of richly contaminated food crops; those are chief part of food chain. Samples of soil, plant, animal blood, hair, and feces were collected to find contamination through wet digestion process in lab and metal analysis. Different forages were collected to study Mn content that was our major concern in this study. The present findings also emphasized on the assessment of bio-concentration factor (BCF). Other significant indices of mobility and pollution of metal were also calculated, i.e. pollution load index (PLI), daily intake of metal (DIM), health risk index (HRI), and enrichment factor (EF). The experimentation result showed different concentrations of metal in different seasons. The Mn concentration in forages was 20.01-28.29 mg/kg and in soil was 5.27-8.90 mg/kg. Soil samples showed higher level of (PLI) Pollution load index. Bio-concentration of MN was 2.59-4.21 mg/kg. It can be concluded that regular monitoring of the metal is essential to evaluate the contamination status. Mn contents were in the safe limits in soil and plants; however, its toxic level was observed in animals., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

This investigation was done for the assurance of potassium amassing in four assortments of maize (grains, shoot and root), soil, and water and in seven tissues of chickens (kidney, liver, heart, bone, gizzard, breast meat). The analysis of variance showed significant differences for potassium concentration in water in all sources of water; however, the season and variety significantly influenced the quantity of potassium in cereals. The corn varieties MMRI, Sadaf, and Pearl behaved differently when treated with water from various sources. Water taken from sewage had a higher concentration of potassium compared to canal and groundwater that is why the maize plants irrigated with this water had a higher grouping. Data regarding potassium concentration in different body parts of chicken showed that season and treatment have a significant effect on the potassium concentration in chicken organs. The variety was non-significant for the potassium concentration only in the bone. Season × Variety interaction was only significant in blood, meat heart, and gizzard. Season × Treatment and Variety × Treatment interactions were significant in the heart, kidney, and gizzard. The potassium contents were higher in the chicken body parts that were reared on grains irrigated with sewage water as compared to other groups. The potassium contents were higher in the chicken meat (96.23 ± 0.00) reared on grains of the Pearl variety raised with the sewage water. In a nutshell, the irrigation of grains with sewage water led to accumulation of nutrients greater than those irrigated with ground or canal water., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

Minerals are essential for ruminants affecting significantly the production of grazing livestock. Iron level in forages, soil, and blood plasma of the small ruminants (goat and sheep) was investigated in three districts of Punjab. Atomic absorption spectrophotometer was used to determine the concentration of iron in collected samples. The results revealed that the mean Fe concentrations in soil of districts Sargodha, Mianwali, and Bhakhar were significantly varied and ranged from 21.85 to 23.78, 28.45 to 31.2, and 18.079 to 24.33 mg/kg, respectively. The Fe level in soil of Mianwali significantly varied and was higher than Sargodha and Bhakkar. The mean Fe concentration in forages which were used for feeding purpose were significantly varied and found between 10.95-14.49, 23.63-25.65, and 6.616-9.45 mg/kg for Sargodha, Mianwali, and Bhakhar, respectively. The mean Fe concentrations in blood plasma of goat which consumed the contaminated forages were 8.5026-11.763 mg/L in district Sargodha, 19.77-20.19 mg/L in Mianwali, and 5.508-5.858mg/L in Bhakkar. In blood plasma of sheep, the residual levels of Fe in districts Sargodha, Mianwali, and Bhakhar were ranged from 9.987 to 12.455, 15.8 to 19.785, and 3.425 to 6.383 mg/L, respectively. This study provides the data of metals effected by different sites and also their mobility from low to higher trophic level which enables us to study the iron toxicity in different trophic levels, and we recommend different safe limits and treatment in case of low and high metal profile., (© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)

The instant endeavor was undertaken for determination of lead (Pb) in water, soil, forage, and cow's blood domesticated in contaminated area of heavy automobiles' exhaust in Sahiwal town of District Sargodha, Pakistan. Water samples showed that the concentration of Pb ranged from 1.14 to 0.44 mg kg -1 at all sites. It was maximum at site 5 and minimum at site 2. Soil samples showed the concentration of Pb at all sites ranged from 1.58 to 0.279 mg kg -1 . It was maximum in soil where Avena sativa was grown at site 5 and was found minimum in soil where Zea mays was grown at site 2. While among samples of forage, the concentration of Pb ranges from 0.048 to 2.002 mg kg -1 . The highest Pb amount was found in Brassica campestris at site 1 and the minimum was recorded in Trifolium alexandrinum at site 2. Finally, the blood samples of cow depicted that concentration of Pb ranged from 4.468 to 0.217 mg kg -1 . It was the maximum at site 1 and the minimum at site 3. It is recommended that such study should be conducted in other districts for public awareness., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)