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Sammar Fathy Elhabal,1 Saeed AS Al-Zuhairy,2 Mohamed El-Nabarawi,3 Mohamed Fathi Mohamed Elrefai,4,5 Mai S Shoela,6 Sandra Hababeh,7 Jakline Nelson,8 Mohamed A Abdel Khalek,9 Marwa Fady,10,11 Nahla A Elzohairy,11,12 Mariam E Amin,13 Gehad M Khamis,6 Amira Rizk,14 Sara Mohamed Ahmed,15 Ahmed A El-Rashedy,16 Mohamed Mohany,17 Abdulaziz S Al-Roujayee,18 Ahmed Mohsen Faheem,19 Amr Amin20 1Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Modern University for Technology and Information (MTI), Mokattam, Cairo, Egypt; 2Department of Pharmacy, Kut University College, Kut, Wasit, Iraq; 3Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 4Department of Anatomy, physiology and Biochemistry, Faculty of Medicine, the Hashemite University, Zarqa, Jordan; 5Department of Anatomy and Embryology, Faculty of Medicine, Ain Shams University, Cairo, Egypt; 6Department of Clinical Pharmacology, Faculty of Medicine, Alexandria University, Alexandria, Egypt; 7Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia; 8Department of Microbiology and Immunology, Faculty of Pharmacy, Nahda University, Beni-Suef, Egypt; 9Institute of Nanoscience and Nanotechnology, Kafrelsheikh University, Kafrelsheikh, 33516, Egypt; 10Zagazig University Hospitals, Infection Control Unit, Zagazig, 44519, Egypt; 11Modern University for Technology & Information, Department of Microbiology and Immunology, Pharmacy College., Cairo Governorat, Egypt; 12Air Force Specialized Hospital, Cairo, Egypt; 13Microbiology and Immunology Department, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt; 14Food Science and technology, Department Faculty of Agricultural, Tanta University, Tanta, Egypt; 15Department of Pharmaceutics, College of Pharmaceutical Sciences and Drug Manufacturing, Misr University for Science and Technology, Giza, 12585, Egypt; 16Chemistry of Natural and Microbial Products Department, National Research Center (NRC), Giza, Egypt; 17Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia; 18Department of Dermatology and Venereology, College of Medicine, Al Imam Mohammad Ibn Saud Islamic University, Riyadh, Saudi Arabia; 19Department of Medical Biochemistry and Molecular Biology, Faculty of Medicine, Mansoura University, Mansoura, Egypt; 20College of Medicine, Sharjah University, Sharjah, United Arab EmiratesCorrespondence: Sammar Fathy Elhabal; Amr Amin, Email sammar_fathy2007@yahoo.com, sammar.fathy@pharm.mti.edu.eg; a.amin@sharjah.ac.aeBackground: Diabetic foot ulcers present a formidable challenge due to colonization by biofilm-forming microorganisms, heightened oxidative stress, and continuous wound maceration caused by excessive exudation.Methods: To address these issues, we developed a robust, stretchable, electro-conductive, self-healing, antioxidant, and antibiofilm hydrogel. This hydrogel was synthesized through the crosslinking of polyvinyl alcohol (PVA) and chitosan (CH) with boric acid. To enhance its antimicrobial efficacy, graphene oxide (GO), produced via electrochemical exfoliation in a zinc ion-based electrolyte medium, was incorporated. For optimal antibiofilm performance, GO was functionalized with cranberry (CR) phenolic extracts, forming a graphene oxide-cranberry nanohybrid (GO-CR).Results: The incorporation of GO-CR into the hydrogel significantly improved its stretchability (280% for PVA/CH/GO-CR compared to 200% for PVA/CH). Additionally, the hydrogel demonstrated efficient photothermal conversion under near-infrared (NIR) light, enabling dynamic exudate removal, which is expected to minimize retained exudate between the wound and the dressing, reducing the risk of wound maceration. The hydrogel effectively reduced levels of lipopolysaccharide (LPS)-induced skin inflammation markers, significantly lowering the expression of NLRP3, TNF-α, IL-6, and IL-1β by 39.2%, 31.9%, 41%, and 52.3%, respectively. Histopathological and immunohistochemical analyses further confirmed reduced inflammation and enhanced wound healing.Conclusion: The PVA/CH/GO-CR hydrogel exhibits multifunctional properties that enhance wound healing ulcers. Its superior mechanical, antibacterial, and anti-inflammatory properties and ability to promote angiogenesis make it a promising candidate for effective wound management in diabetic patients. Keywords: antibiofilm, cranberry extract, electro conductive hydrogel, graphene oxide, nanocomposite, photothermal conversion, self-healing, wound healing

The current study aimed to assess the neuroprotective effect of Zn and Fe oxide nanoparticles biofabricated by Taverniera glabra in alloxan-induced diabetic rats. The experimental animals (160–200 g) were divided into nine groups (n = 9). The blood glucose, body weight, glucose-6-phosphate dehydrogenase (G6PD), superoxide dismutase (SOD), catalase test (CAT), lipid peroxidation (TBARS), glutathione (GSH), and acetylcholinesterase (AChE) activities were determined. Oral administration of nanoparticles and T. glabra methanol extract (TGME; 10 and 15 mg/kg b.w) significantly decreased the glucose level, increased the body weight, controlled the quantitative level of G6PD, and significantly decreased the levels of ALT, ALP, cholesterol, and creatinine. Moreover, TGME and their Zn and Fe oxide nanoparticles significantly restored the antioxidant enzymes (SOD, CAT, GPx, and GSH) that decreased during induced diabetes. In the diabetic group, a significant increase in TBARS was noted and recovered in diabetic animals (p < 0.05) as compared to glibenclamide. The AChE activity was significantly recovered by nanoparticles and TGME both in the blood and brain of the diabetic group (p < 0.05). Taken together, it can be suggested that TGME and Zn and Fe oxide nanoparticles significantly improved memory and could be considered as an effective biogenic nanomaterial for diabetes, Alzheimer’s disease, and oxidative stress.

Cancer is the second largest cause of mortality worldwide. Many natural bioactive chemicals generated from plants have favorable impacts on health, including cancer chemoprevention, compared to their manufactured counterparts. Persicaline, a novel sulfur-containing imidazoline alkaloid derived from Salvadora persica, has been shown to display promising antioxidant activity. In this study, the antiproliferative activity of persicaline was tested against different cancer cells using (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay. The cell death mode and cell-cycle arrest were examined using flow cytometry analysis. In addition, the proapoptotic and molecular mechanism effects of persicaline against mammary MCF-7 cell line were explored. Furthermore, the impact of persicaline on apoptotic genes markers, generation of reactive oxygen species (ROS), and mitochondrial membrane potential were monitored. It was found that persicaline inhibits cell proliferation in a dose-dependent manner. Persicaline-treated MCF-7 cells also showed initiation of apoptotic events and G1 cell-cycle arrest. In addition, persicaline treatment led to an increase in ROS generation, Bax and caspase upregulation while the Bcl-2 was downregulated. Hence, for the first time, this study showed that persicaline causes G1 phase arrest and apoptosis induction in MCF-7 cells. Increased proapoptotic genes and ROS levels were required for the antiproliferative and apoptotic effects of persicaline.

Abstract Background Continuous diabetes-associated complications are a major source of immune system exhaustion and an increased incidence of infection. Diabetes can cause poor circulation in the feet, increasing the likelihood of ulcers forming when the skin is damaged and slowing the healing of the ulcers. Whey proteins (WPs) enhance immunity during childhood and have a protective effect on some immune disorders. Therefore, in this study, we investigated the effects of camel WP on the healing and closure of diabetic wounds in a streptozotocin (STZ)-induced type I diabetic mouse model. Results Diabetic mice exhibited delayed wound closure characterized by a significant decrease in an anti-inflammatory cytokine (namely, IL-10) and a prolonged elevation of the levels of inflammatory cytokines (TNF-α, IL-1β and IL-6) in wound tissue. Moreover, aberrant expression of chemokines that regulate wound healing (MIP-1α, MIP-2, KC and CX3CL1) and growth factors (TGF-β) were observed in the wound tissue of diabetic mice compared with control nondiabetic mice. Interestingly, compared with untreated diabetic mice, supplementation with WP significantly accelerated the closure of diabetic wounds by limiting inflammatory stimuli via the restoration of normal IL-10, TNF-α, IL-1β and IL-6 levels. Most importantly, the supplementation of diabetic mice with WP significantly modulated the expression of MIP-1α, MIP-2, KC, CX3CL1 and TGF-β in wound tissue compared with untreated diabetic mice. Conclusion Our data demonstrate the benefits of WP supplementation for improving the healing and closure of diabetic wounds and restoring the immune response in diabetic mice.

Abstract Background Thymoquinone (TQ), the major active component of the medicinal herb Nigella sativa Linn., has been described as a chemopreventive and chemotherapeutic compound. Methods In this study, we investigated the effect of TQ on survival, actin cytoskeletal reorganization, proliferation and signal transduction in multiple myeloma (MM) cells. Results We found that TQ induces growth arrest in both MDN and XG2 cells in a dose- and time-dependent manner. TQ also inhibited CXC ligand-12 (CXCL-12)-mediated actin polymerization and cellular proliferation, as shown by flow cytometry. The signal transducer and activator of transcription (STAT) and B-cell lymphoma-2 (Bcl-2) signaling pathways may play important roles in the malignant transformation of a number of human malignancies. The constitutive activation of the STAT3 and Bcl-2 pathways is frequently observed in several cancer cell lines, including MM cells. Using flow cytometry, we found that TQ markedly decreased STAT3 phosphorylation and Bcl-2 and Bcl-XL expression without modulating STAT5 phosphorylation in MM cells. Using western blotting, we confirmed the inhibitory effect of TQ on STAT3 phosphorylation and Bcl-2 and Bcl-XL expression. Conclusions Taken together, our data suggests that TQ could potentially be applied toward the treatment of MM and other malignancies.

Abstract Background Long and persistent uncontrolled diabetes tends to degenerate the immune system and leads to an increased incidence of infection. Whey proteins (WPs) enhance immunity during early life and have a protective role in some immune disorders. In this study, the effects of camel WP on the chemotaxis of B and T cells to CXCL12 and CCL21 in diabetic mice were investigated. Results Flow cytometric analysis of the surface expressions of CXCR4 (CXCL12 receptor) and CCR7 (CCL21 receptor) on B and T cells revealed that the surface expressions of CXCR4 and CCR7 were not significantly altered in diabetic and WP-supplemented diabetic mice compared with control mice. Nevertheless, B and T lymphocytes from diabetic mice were found to be in a stunned state, with a marked and significant (P < 0.05) decrease in CXCL12- and CCL21-mediated actin polymerization and subsequently, a marked decrease in their chemotaxis. WP supplementation in the diabetes model was found to significantly increase CXCL12- and CCL21-mediated actin polymerization and chemotaxis in both B and T cells. Conclusion Our data revealed the benefits of WP supplementation in enhancing cytoskeletal rearrangement and chemotaxis in B and T cells, and subsequently improving the immune response in diabetic mice.

Heavy metal bioaccumulation in organisms is primarily a result of dietary uptake. The current study examines the concentrations of heavy metals (Pb, Cd, Cr, and Cu) in fish feed, water, sediment, and three fish species (Catla catla, Labeo rohita, and Cyprinus carpio) from different feeding zones in a polyculture pond system. Furthermore, associated human health risks were also evaluated. The fish samples (n = 25 for each species) were collected from 10 different fish ponds in the Kohat district, Pakistan. Heavy metals were determined using an atomic absorption spectrometer. Results revealed higher concentrations of heavy metals in sediment, followed by water. However, the concentration of heavy metals in fish feed was lower than the standard limits. In the case of fish, the bottom feeder (C. carpio) notably exhibited higher (P < 0.05) levels of heavy metals than the column feeder (L. rohita) and surface feeder (C. catla) fish. Moreover, in the liver of all fish species, the bioaccumulation of heavy metals was higher, followed by the gills. Principal component analysis (PCA) demonstrated a strong correlation of heavy metals in C. carpio gills, flesh, feed, and pond water, while the heavy metals in the liver correlated with the detected metals in sediment. The human health risk analysis shows that bottom feeder fish had higher estimated daily intake (EDI), target hazard quotient (THQ), and hazard index (HI) values (> 1). Consequently, the exposed population may experience adverse health effects. The findings of this study suggest that the bottom feeder (C. carpio) bioaccumulates a higher concentration of heavy metals than column (L. rohita) and surface feeder (C. catla) in the polyculture system., Competing Interests: Declarations. Ethics Approval: The authors followed all the valid national rules for the use and care of animals, and the study was conducted after the ethical committee approval of the Department of Zoology, University of Lahore, Pakistan. Consent for Publication: Not applicable. Competing Interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background : in the twenty-first century, the emergence of COVID-19 as a highly transmissible pandemic disease caused by SARS-CoV-2 posed a significant threat to humanity. Aims & Objectives : the disease spreads through small respiratory droplets, necessitating the use of various compounds for treatment, with alkaloids being recognized as particularly crucial owing to their diverse pharmaceutical properties. Methodology : in this study, a dataset comprising 100 natural alkaloids obtained from the literature was transformed into 2D chemical structures using Chem Draw 19.1. Subsequently, 3DQSAR studies were conducted on the dataset, resulting in the automatic screening of 50 compounds from the initial pool of 100 compounds. The values of q 2 and r 2 of the validated field-based 3DQSAR model were 0.7186 and 0.971, respectively. The validated atom-based 3DQSAR model has q 2 and r 2 scores of 0.6025 and 0.9845, respectively. Based on the obtained results, 10 compounds with exceptionally active predictive IC 50 values were selected for further analysis. Docking experiments were then performed on the selected compounds, and the top three compounds with the highest docking scores were identified as diazepinomicin, (+)- N -methylisococlaurine, and hymenocardine-H. After docking, MM-GBSA was performed on the complexes of diazepinomicin, (+)- N -methylisococlaurine and hymenocardine-H with their corresponding proteins, which resulted in the authentication of the molecular docking scores. MD simulations were also performed to check the flexibility, stability and compactness of these complexes for revalidation of docking scores. Results : finally, ADMET experiments revealed that (+)- N -methylisococlaurine exhibited the most favourable properties among these three compounds., Competing Interests: Authors have no conflict of interest., (This journal is © The Royal Society of Chemistry.)

Nanomedical applications have increased significantly. This work aimed to fabricate and characterize cobalt oxide nanoparticles (CoOnps) synthesized biologically via aqueous Alhagi maurorum extract and evaluate their cytotoxic and antimicrobial impacts. Green-synthesized CoOnps were prepared and analyzed using UV-Vis spectrophotometer UV-vis, Scanning electron microscopy (SEM), Transmission electron microscopy TEM, Energy dispersive X-ray analysis EDAX, Fourier transform infrared, FTIR, and X-ray diffraction (XRD). In vitro traits of green-synthesized CoOnps were studied on ovarian cancer cells (SKOV3) using a Sulforhodamine B (SRB) method. The cytotoxic effect and IC50 were estimated. Moreover, concentrations of 10, 30, 40, 70, 100, 200, 300, 400 and 500 μg/mL CoOnps were applied to investigate their antimicrobial effect against Listeria, Staphylococcus aureus and Streptococcus as gram +ve pathogenic bacteria, Bifidobacterium bifidum 2203, Bifidobacterium bifidum LMG 10,645, Bifidobacterium breve LMC 017, Bifidobacterium angulatum 2238 and Bifidobacterium longum ATCC 15,707 as probiotics, E. coli as gram -ve bacterial model and yeast strain Candida albicans. CoOnps showed anti-ovarian cancer effects at 24.02 μg/mL. Furthermore, it exerted antimicrobial activity versus Listeria, Streptococcus, S. aureus, and E. coli were 31.66 ± 0.88, 24.33 ± 2.08, 25.66 ± 0.33, and 33.00 ± 6.08; however, they did not suppress the growth of Candida albicans and all tested Bifidobacterial strains up to concentrations of 500 μg/mL with significant difference compared to all concentrations p < 0.05. Green synthesis of CoOnps is a low-cost, eco-friendly and easily prepared method. Its impressive features as cytotoxic SKOV3, a cell line ovarian cancer and antibacterial effect for some gram +ve and -ve bacteria, besides maintaining probiotics, could candidate them as competitive agents for medical, pharmacological, agricultural and food applications., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Compliance with ethics guidelines: Ethical approval no MU/FA 0060223., (© 2024. The Author(s).)

Extended spectrum β -lactamase (ESBL)-producing Enterobacteriaceae, including Klebsiella pneumoniae and Escherichia coli , pose a serious risk to human health because of antibiotic resistance. Wastewater serves as a reservoir for these bacteria, contributing to the evolution and transmission of antibiotic-resistant strains. The research aims to identify ESBL bacterium in wastewater samples from District Kohat. K. pneumoniae and E. coli were confirmed as ESBL-producing bacteria through a comprehensive array of diagnostic procedures, including Gram staining, biochemical analyses, and antibiotic susceptibility testing. Fecal coliform count (FCC) analyses revealed varying microorganism levels. Both E. coli and K. pneumoniae isolates showed ESBL enzyme expression, indicating antibiotic resistance. Resistance patterns included ciprofloxacin, ampicillin, cefotaxime, cefoxitin, and amoxicillin-clavulanic acid for both species. E. coli displayed higher sensitivity for chloramphenicol, trimethoprim- sulfamethoxazole, and gentamicin. Ceftazidime minimum inhibitory concentration results showed E. coli 's higher resistance. The study accentuates the presence of antibiotic-resistant strains, emphasizing the value of effective wastewater treatment. The study provides crucial insights into microbial characteristics, fecal contamination, ESBL production, and antibiotic resistance in E. coli and K. pneumoniae isolates, advocating for monitoring and mitigation strategies., Competing Interests: The authors declare there is no conflict., (© 2025 The Authors This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC BY 4.0), which permits copying, adaptation and redistribution, provided the original work is properly cited (http://creativecommons.org/licenses/by/4.0/).)

The exposure of fish to heavy metals can significantly impact physiological processes and potentially lead to adverse health effects. This study assesses the effects of exposure to Cd and Pb sublethal concentrations in water on Wallagu attu. A total of 48 fish with an average body weight of 145.5 ± 26 g were distributed among three groups (control, Cd-treated, and Pb-treated) within 60 L fiberglass tanks. They were exposed to 30% sublethal concentrations of Cd and Pb for durations of 1, 15, and 30 days. Following this exposure, an assessment was conducted on metal bioaccumulation and hemato-biochemical responses. Results revealed a significantly (P < 0.05) higher concentration of heavy metals in the fish tissues of metals exposed groups than in the control. The concentration of Cd and Pb increases in fish tissues (kidney > gills > intestine) with exposure time. In most cases, the Pb-exposed group exhibited significantly (P < 0.05) higher concentrations of Pb in different tissues than the Cd-treated group. With extended exposure time, the activities of CAT and SOD show a significant decrease in both Cd and Pb-treated groups. However, the reduction in activities was more pronounced in the Cd-exposed group. On 15 and 30 days, the levels of red blood cells (RBC), hemoglobin (HB), hematocrit (HCT), and total protein (TP) decrease in groups exposed to Cd and Pb. The cortisol and glucose levels exhibit a more noticeable (P < 0.05) increase with prolonged exposure to Cd and Pb than the control group. On day 30, the survival rate decreased more in the Pb-exposed group. The findings of this study indicate that exposure to sublethal doses of Cd and Pb induces stress in Wallagu attu, resulting in rapid changes in specific hemato-biochemical parameters., Competing Interests: Declarations. Ethical Approval: The authors adhered to all relevant national regulations governing the ethical treatment and use of animals, and the study was carried out following approval from the Zoology Department, University of Sargodha ethical committee. Consent for Publication: Not applicable. Competing Interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Human activities have far-reaching impact on natural ecosystems, causing increasing disturbances and disruptions to the delicate balance of the environment. Poor land use planning, urbanization, infrastructure development, and unplanned tourism exacerbate contamination and degradation in tourist destinations, yet the pollution of potentially toxic elements (PTEs) in these environments remains inadequately explored. To address this issue, we investigated the concentrations of acid-digested PTEs in road dust in Abbottabad city (Pakistan) with heavy traffic. The current study also evaluated PTE associated pollution, ecological and health risks, and potential sources of these elements. The average concentrations (in mg kg -1 ) of PTEs in road dust were highest for Fe (15540), followed by Mn (304), Zn (139), Cu (50.0), Pb (21.5), Cr (13.0), Ni (10.3), Co (6.66), and Cd (0.236). The levels of Co, Cr, Mn, Ni, and Fe were below the upper continental crust (UCC) averages, while Cd, Cu, Pb, and Zn exceeded them. Spatially, Cd, Cu, Pb, and Zn were found at higher levels near traffic hotspots, bus stands and automobile workshops. Road dust in Abbottabad exhibited unpolluted to moderate pollution levels (geo-accumulation index), with Cd, Cu, Zn, and Pb at 23 % of the sites. The enrichment factor results indicated a significant anthropogenic influence, with Cd being significantly enriched and Zn, Cu, and Pb moderately enriched. The contamination factor results revealed moderate contamination by (Cd: 2.62, Zn: 2.08, Cu: 1.79, Pb: 1.27). Single metal risk index showed that 61 % of the sites posed considerable to very high ecological risks due to Cd, which was highlighted as a significant concern. The absolute principal component scores-multiple linear regression model identified three factors contributing to PTE pollution: lithogenic (Co, Fe, Mn, Ni), traffic-related (Cd, Cu, Pb, Zn), and mixed sources (Cr), with contributions of 52.8 %, 35.8 %, and 11.4 %, respectively. The PTE hazard quotient and total hazard index values for children and adults were below the safe risk level of 1, indicating no non-carcinogenic health risks. The cumulative carcinogenic risk values to the residents were also within acceptable limits. However, children's susceptibility to non-carcinogenic risks is higher due to their behavior and lower body weight. This study highlights the accumulation of PTEs in Abbottabad's environment, which poses challenges to long-term sustainability, particularly amid unplanned tourism. Therefore, urgent measures are necessary to mitigate PTE pollution and preserve tourist spots and public health., 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 Elsevier B.V. All rights reserved.)

Background: Heavy metal pollution, particularly lead (Pb), poses a significant threat to aquatic ecosystems and their inhabitants, threatening their delicate balance and long-term viability. This study highlights the urgent need to mitigate heavy metal pollution in aquatic ecosystems., Objective: This study investigates Pb(NO 3 ) 2 exposure effects on tissue accumulation, behavioral abnormalities, and hemato-biochemical parameters in common carp (Cyprinus carpio), a widely distributed freshwater fish species., Methodology: Fish (115 ± 5.23 g) were exposed to various Pb(NO 3 ) 2 concentrations for 10 and 20 days, representing control (0 %), 25 %, 50 %, and 75 % of the LC 50 equivalent to 19.33, 38.66, and 58.0 mg/l, respectively. The standard manual procedure was used for blood sampling. The lead concentration in fish tissue was determined using an atomic absorption spectrophotometer., Results: Results revealed that fish gills showed significant (P < 0.05) increase in Pb(NO 3 ) 2 after 10 days, further rising after 20 days. Liver concentrations also rose significantly (P < 0.05) with prolonged exposure and increasing Pb levels. Muscle had lower concentrations. Hematological parameters (RBC, WBC, HB, HCT) decreased with higher Pb(NO 3 ) 2 levels. Behavioral and morphological changes were significantly more pronounced in the exposure groups when compared to the control group. Hepatic enzyme activities (AST, ALT), glucose, and lipid levels increased, while total protein decreased., Conclusions: The study highlights Pb(NO 3 ) 2 harmful effects on common carp, impacting tissue accumulation, hematological parameters, and biochemical disruptions. It emphasizes the need to monitor and mitigate heavy metal pollution in aquatic environments to safeguard freshwater organisms and ecosystems, and to further increase our understanding of Pb toxicity in freshwater ecosystems., 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. Competing Interests The authors declare that they have no competing interests., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

The significance of plant-derived products in aquaculture lies in their potential to offer sustainable alternatives, promoting eco-friendly practices. This study investigated the impact of nettle (Urtica dioica) leaves powder on the growth efficiency, hemato-biochemical variables and non-specific immune system of rohu, Labeo rohita fingerlings. To achieve this objective, sample average weight (5.23 ± 0.34 g) were categorized into four groups, namely control, T1, T2 and T3 in triplicate and administered diets fortified with nettle in amounts of 0, 1, 3 and 5% respectively for a duration of 60 days. After 60 days of fortified diet, the fish underwent intraperitoneal injection with bacteria (Aeromonas hydrophila), and subsequent relative percentage survival (RPS) was observed. The growth performance, including "weight gain (WG), specific growth rate (SGR), feed conversion ratio (FCR) and feed efficiency ratio (FER)", were notably higher in the T3 group (5%) than in others. The hematological values of White blood cell, hematocrit, and hemoglobin revealed higher levels with a fortified diet. The dietary supplementation of nettle reduced serum cholesterol and glucose concentration, whereas it increased albumin, globulin, and total protein in the fish blood. Enhancements in lysozyme and myeloperoxidase activity were observed in the intervention groups with feed containing nettle supplementation. The nettle diet at a 5% concentration demonstrated a higher RPS than the others following injection with A. hydrophila. The findings indicate the potential of nettle as a valuable nutritional supplement for increasing fish immunological reaction and bolstering pathogen resistance., Competing Interests: Declarations Informed consent Not applicable. Institutional Review Board The research was reviewed and approved by the Advanced Research Board of the Department of Zoology, Khushal Khan Khattak University. All procedures conducted in the study were also approved by the Ethics Committee of Khushal Khan Khattak University, Karak, Pakistan (registration number: Zoo/KKKU/13). Competing interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Murideoxycholic acid (MDCA), as a significant secondary bile acid derived from the metabolism of α/β-muricholic acid in rodents, is an important component in maintaining the bile acid homeostasis. However, the biosynthesis of MDCA remains a challenging task. Here, we present the development of cytochrome P450 monooxygenase CYP102A1 (P450 BM3) from Bacillus megaterium, employing semi-rational protein engineering technique. Following three rounds of mutagenesis, a triple variant (T260G/G328A/L82V) has been discovered that proficiently catalyzes the 6β-hydroxylation of lithocholic acid (LCA), thereby generating MDCA with an impressive 8.5-fold increase in yield compared to the template P450 BM3 mutant. The MDCA selectivity has been also promoted from 62.0% to 96.3%. This biocatalyst introduces a novel approach for the biosynthesis of MDCA from LCA. Furthermore, molecular docking and dynamics simulations have been employed to unravel the molecular mechanisms underlying the enhanced LCA conversion and MDCA selectivity., (© 2024 Wiley‐VCH GmbH.)

Background: In China, decline disease with unknown etiology appeared as an epidemic among bayberry trees in the southern area of the Yangtze River. Furthermore, the use of beneficial microbes has been reported to be able to reduce the incidence of this disease, emphasizing the association of this disease with microorganisms. Therefore, it has become critical to uncover the microbiome's function and related metabolites in remodeling the immunity of bayberry trees under biotic or abiotic stresses., Results: The amplicon sequencing data revealed that decline disease significantly altered bacterial and fungal communities, and their metabolites in the four distinct niches, especially in the rhizosphere soils and roots. Furthermore, the microbial communities in the four niches correlated with the metabolites of the corresponding niches of bayberry plants, and the fungal and bacterial networks of healthy trees were shown to be more complex than those of diseased trees. In addition, the role of microbiome in the resistance of bayberry trees to the occurrence of decline disease was justified by the isolation, identification, and characterization of important microorganisms such as significantly enriched Bacillus ASV804, Pseudomonas ASV815 in healthy plants, and significantly enriched Stenotrophomonas ASV719 in diseased plants., Conclusion: Overall, our study revealed that the occurrence of decline disease altered the microbiome and its metabolites in four ecological niches in particular rhizosphere soils and roots of bayberry, which provides new insight into the control of bayberry decline disease., (© 2024. The Author(s).)

Backgroud: Pregnancy is a state of increased metabolic demand, and poor maternal nutrition can exacerbate these changes, potentially increasing the risk of cardiometabolic disorders.This study is significant as it investigates the effect of high energy nutritional supplements on cardio metabolic risk markers in underweight primigravidas., Methodology: A single blinded randomized controlled trial was conducted from April 2018- August 2019 among the Pakistani tertiary care facilities of Khyber Pakhtunkhwa. A total of thirty six participants were randomized into two groups receiving either high energy nutritional supplement 'MAAMTA' or Placebo. They were instructed to take Placebo/Supplement alongside to their regular prenatal care and food from their first antenatal visit till a week postnatally. Anthropometric measurements and blood samples for biochemical parameters insulinlevels, fasting blood glucose & lipid profile were taken at baseline visit, between 16 and 20 weeks gestation and a post natal visit., Results: The two-way repeated measures ANOVA showed that there was a highly significant time effect on participant's Insulin level (F (1.8, 53.6) = 10.64, P < 0 0.000, ηp 2  = 0.269). A highly significant time effect on participant's insulin resistance (F (2, 68) = 8.116,P < 0 0.001, ηp 2  = 0.193) was also observed. There was a highly significant time effect on participant's LDL level as well (F (2, 68) = 11.82, P < 0 0.000, ηp 2  = 0.258)., Conclusion: Supplementation with high energy nutritional supplements may improve insulin levels and insulin sensitivity in underweight primigravidas., Trial Registration: https://www., Clinicaltrials: gov ClinicalTrials.gov Identifier: ISRCTN 10088578. Registered on 27 March 2018. https://www.isrctn.com/ISRCTN10088578 ., (© 2024. The Author(s).)

Over-fertilizing fish ponds can cause pollution, introducing heavy metals into the food chain and posing health risks. The present study investigated the incidence of heavy metals (Pb, Cu, Cd, and Cr) in commonly applied fertilizers, including nitrogen, phosphorus and potassium (NPK), triple superphosphate (TSP), and di-ammonium phosphate (DAP), and their association with heavy metals in water, sediment, and cultured fish species (Catla catla, Labeo rohita, and Cyprinus carpio) in polyculture fish ponds. The study was conducted over 4 months, with four groups in triplicates: control (no fertilizer), group 1 (NPK), group 2 (TSP), and group 3 (DAP). Heavy metal analysis was carried out using atomic absorption spectrophotometry before and after fertilizer application. Significantly (p < 0.05) higher levels of heavy metals were observed in water and sediment after applying fertilizers, with the most pronounced results in group 3 (DAP) followed by group 2 (TSP). The concentration of heavy metals was significantly (p < 0.05) higher in group 3 (DAP) fertilizers compared to other groups. Compared to the control, the concentration and bioaccumulation of heavy metals were significantly (p < 0.05) higher in the fertilizer-applied groups, with notably higher levels in group 3 (DAP). Cluster analysis and the correlation matrix did not show any significant association between the heavy metals and the fertilizers, indicating a complex interplay between the biotic and abiotic factors of the system. The health index (HI) value was < 1 in fish muscles of all studied groups, indicating the fish are safe for consumption. The study recommends monitoring and regulating fertilizer use, especially DAP, to prevent heavy metal contamination, and exploring sustainable alternatives to minimize environmental and health risks., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

There has been increasing interest in biologically active plant extracts. Studies continue to discover novel components, especially those with anti-anxiety activities. The present study investigates the anxiolytic activity of Salvia moorcroftiana Wall. ex Benth. aerial parts through both in vivo and in silico studies. Aerial parts of the experimental plant were extracted using a hydroalcoholic solvent and fractionated with various organic solvents of differing polarities, including hexane, dichloromethane, ethyl acetate, and n-butanol. The chemical compositions were determined using gas chromatography-mass spectrometry (GC/MS). In vivo anti-anxiety activity was tested on various Swiss albino mice models. Results indicate that all fractions of S. moorcroftiana exhibited significant anxiolytic effects, with the butanol fraction displaying the highest efficacy. Molecular docking analysis suggested that some of the compounds could target anxiety disorder proteins. ADME/T calculations were performed to examine the effects of S. moorcroftiana extracts on human metabolism. Therefore, the present study establishes the significant anti-anxiety activity of S. moorcroftiana aerial parts, suggesting its potential as a therapeutic agent for various anxiety disorders.

This study focused on analyzing the pharmacological activities of AgNPs synthesized from an aqueous plant extract of Anchusa arvensis . The effectiveness of AgNPs was evaluated for protein kinase inhibition, antioxidant, antibacterial, and antifungal activities. The AgNPs and plant were used to regulate the protein kinase activity using the liquid TSB and ISP4 medium protein kinase inhibition study demonstrated that nanoparticles exhibited a larger zone of inhibition (9.1 ± 0.8) compared to the plant extract (8.1 ± 0.6). The antioxidant activity was assessed using DPPH reagent, and the results indicated that AgNPs displayed potent free radical scavenging properties. In terms of antibacterial activity, AgNPs showed higher efficacy against Enterobacter aerogens (20.1 ± 0.9), Bordetella bronchiseptaca (19.1 ± 0.9), and Salmonella typhimurium (17.2 ± 0.8) at 4 mg/mL. The antifungal activity of AgNPs was prominent against Aspergillus fumagatus (14.1 ± 0.9), Mucor species (19.2 ± 0.8), and Fusarium solani (11.2 ± 0.8) at 20 mg/mL. These findings suggest that AgNPs possess multiple beneficial properties, including bactericidal/fungicidal effects, protein kinase inhibition, and potential free radical scavenging abilities. Therefore, AgNPs have potential applications in various fields, such as biomedicine and industry, due to their ability to counteract the harmful effects of free radicals., (© 2024 Walter de Gruyter GmbH, Berlin/Boston.)

Salvia officinalis L., commonly known as sage and belonging to the Lamiaceae family, is a medicinal herb indigenous to the Mediterranean region. It is celebrated for its diverse pharmacological properties and traditional uses in folk medicine, particularly in addressing hepatotoxicity. Cisplatin (Cis), a potent chemotherapeutic agent widely employed in cancer treatment, is recognized for its efficacy but often accompanied by adverse effects, including hepatotoxicity. The aim of this study was to assess whether an ethanolic S. officinalis extract (ESOE) could provide protection against Cis-induced hepatotoxicity in an experimental rat model. The ESOE was prepared using standard extraction techniques, and its chemical constituents were elucidated through UPLC-ESI-MS/MS analysis, revealing the presence of bioactive compounds such as alkaloids, phenolic compounds, and flavonoids, which are associated with various therapeutic effects, including hepatoprotection. Adult male albino rats were allocated into four groups: control, ESOE (250 mg/kg), Cis (7.5 mg/kg), and ESOE (250 mg/kg) + Cis (7.5 mg/kg). The treatment duration lasted 21 days, with Cis administration on the 22nd day. Twenty-four hours post-Cis administration, blood and liver samples were collected for analysis. Cis-induced hepatotoxicity was evidenced by alterations in hematological parameters, including erythrocyte, thrombocyte, leukocyte, and lymphocyte counts, alongside elevated serum levels of liver enzymes (ALT, LDH, AST, ALP, and GGT), indicative of liver damage. Furthermore, Cis exposure resulted in increased hepatic malondialdehyde (MDA) and Nitric oxide (NO) levels, oxidative stress markers, coupled with decreased levels of reduced glutathione (GSH), a non-enzymatic antioxidant, and histopathological changes in liver tissue, characterized by necrosis and inflammation. Additionally, Cis treatment led to elevated levels of 8-hydroxy-2'-deoxyguanosine (8-OH-dG), TNF-α, and IL-6, indicating oxidative stress and inflammation. Remarkably, pretreatment with ESOE ameliorated these Cis-induced hepatotoxic effects, as evidenced by improved hematological parameters, reduced liver enzyme activities, alleviated oxidative stress, and ameliorated histopathological alterations. The observed hepatoprotective effects of ESOE against Cis-induced liver injury may be attributed to its antioxidant and anti-inflammatory properties, highlighting its potential as a natural therapeutic agent in mitigating chemotherapy-associated hepatotoxicity., 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 Elsevier Inc. All rights reserved.)

Objectives: To enhance the utilization of reclaimed land, Sanyeqing (SYQ) has been extensively cultivated in Zhejiang province, China. However, the prevalence of anthracnose has significantly hindered SYQ growth, emerging as a primary obstacle to its production. This study aimed to elucidate SYQ's responses to anthracnose in reclaimed land environments by comprehensively analyzing root-zone bacterial community structure, metabolites, and soil properties., Methods: The experiment was conducted on reclaimed land in Chun'an, China. In order to evaluate the responses of SYQ to anthracnose, the fresh and dry weight of SYQ tubes, the soil properties, the high-throughput sequencing, and metabolomics assay were carried out., Results: Significant differences were observed between an anthracnose-resistant variety (A201714) and an anthracnose-susceptibile variety (B201301). Fresh and dry weight increased 131.53 % and 144.82 % for A201714 compared to B201301. Lacibacterium (39.85 %), Gp6 (21.83 %), Gp5 (21.49 %), and Sphingomonas (18.84 %) were more prevalent, whereas Gp3 (22.71 %), WPS-1 (18.88 %), Gp4 (15.60 %), Subdivision3 (14.70 %), Chryseolinea (14.37 %), and Nitrospira (0.76 %) were less prevalent in A201714 than B201301. A total of 24 bacterial biomarkers were detected in all soil samples, while the network suggests a more stable soil bacterial community in A201714 than in B201301. Eight differentially expressed metabolites (DEMs) that belonged to lipids and lipid-like molecules, organic acids and derivatives, benzenoids, nucleosides, nucleotides, and analogues were found between two soil samples, and all these eight DEMs were downregulated in A201714 and had a strong correlation with 12 genera of bacteria. Moreover, the data from the redundancy analysis indicated that the main variables affecting changes in the bacterial communities were pH, available phosphorus (AP), available potassium (AK), microbial biomass carbon (MBC), and microbial biomass nitrogen (MBN)., Conclusion: This research offers new insights into the SYQ response to anthracnose in reclaimed land and provides valuable recommendations for the high-quality SYQ cultivation and production., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Author [Bin Li] is an AE of this journal. Other 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.)

Fusarium root rot caused by the Fusarium species complex significantly affects the yield and quality of Angelica sinensis , a valuable medicinal herb. Traditional management primarily relies on chemical fungicides, which have led to pathogen resistance, environmental hazards, and concerns regarding public health and the active components in A. sinensis . This study explores the efficacy of a novel plant-derived biopesticide Shi Chuang Zhi Feng Ning (T1; SCZFN), alongside Bacillus subtilis wettable powder (T2) and a chemical fungicide (T3), in controlling root rot and understanding their impacts on the rhizosphere microbial community and root metabolome. Results of the field experiment demonstrated that treatments T1 and T3 achieved control efficiencies of 73.17% and 75.45%, respectively, significantly outperforming T2 (39.99%) and the control. High-throughput sequencing revealed that all treatments altered the diversity and structure of microbial communities, with T1 and T2 reducing the abundance of taxa linked to root rot, such as Muribaculaceae spp., Humicola spp., Fusarium spp., and Mycochlamys spp. Treatment T1 notably enhanced beneficial bacterial taxa, including Acidobacteria spp., Nitrospira spp., and Pedosphaeraceae spp., involved in carbon cycling and plant growth promotion. Metabolomic analysis identified 39, 105, and 45 differentially expressed metabolites (DEMs) across the treatments, demonstrating T1's potential to modulate the root metabolome effectively. Further, a correlation analysis demonstrated a stronger correlation between distinct microorganisms with significant influence and DEMs of T1 treatment compared to other treatments. These findings underscore biopesticide SCZFN's role in enhancing plant health and disease suppression in A. sinensis , providing insights into its biocontrol mechanisms and supporting the development of sustainable disease management strategies in its cultivation.

The use of medicinal plants is integral to addressing liver, heart, lung, and other metabolic issues. These plants are rich in vitamins, minerals, flavonoids, and alkaloids, which collectively help in lowering intestinal glucose absorption and increasing insulin secretion by pancreatic tissues. Elemental analysis, encompassing major, minor, and trace elements, was performed on various parts (leaves, roots, and seeds) of 16 anti-diabetic medicinal plants collected from different regions of Swat and Peshawar in Khyber Pakhtunkhwa (KPK), Pakistan. This analysis utilized proton-induced X-ray emission (PIXE) and inductively coupled plasma optical emission spectroscopy (ICP-OES) techniques. Our PIXE and ICP-OES analysis revealed the presence of major (Ca, K, S, P), minor (Si, Cl), and trace (Al, Mn, Fe, Ni, Cu, Zn, Se, Cr, and Sc) elements in various parts (leaves, roots, and seeds) of the 16 anti-diabetic medicinal plants studied. Specifically, elements such as Ca, K, Cr, Cu, Mn, Zn, and Se were detected, all of which are known to contribute in maintaining normal glucose metabolism. Notably, Zn and Se are crucial trace elements for the synthesis, secretion, and action of insulin. Significant Zn concentrations were observed in ten anti-diabetic medicinal plants: Albizia lebbeck (AL), Atropa acuminata (AA), Avena fatua (AF), Citrus medica (CM), Commiphora wightii (CW), Cymbopogon citratus (CC), Daucus carota (DC), Ziziphus mauritiana (FM), Hyoscyamus niger (HN), and Martynia annua (MA), and significant Se concentrations were observed in twelve medicinal plants, i.e., Albizia lebbeck (AL), Allium sativum (AS), Atropa acuminata (AA), Avena fatua (AF), Cannabis sativa (CS), Capparis spinosa (CaS), Commiphora wightii (CW), Cymbopogon citratus (CC), Datura alba (DA), Daucus carota (DC), Ziziphus mauritiana (FM), and Hyoscyamus niger (HN). Our study's elemental analysis using PIXE and ICP-OES on various parts of 16 medicinal plants identified a significant number of useful elements. Elements such as Ca, K, S, P, Al, Si, Cl, Mn, Fe, Ni, Cu, Zn, Se, and Cr were identified and quantified. These findings support the potential use of these plants in managing diabetes and highlight the importance of elemental profiling in understanding their therapeutic properties., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

This study examines the levels of heavy metals in polyculture fish (Labeo rohita, Cyprinus carpio, and Catla catla), water, and sediment in Tanda Dam, Kohat, Pakistan, aiming to understand environmental and health risks. Samples of fish, water, and sediment were collected from 3 fish farms, and heavy metal concentrations were measured using a Flame Atomic Absorption Spectrophotometer (AAS). Results reveal that C. catla exhibited significantly higher (p < 0.05) levels of Zn than other fish species. Conversely, C. carpio showed significantly higher (p < 0.05) concentrations of Pb, Cd, Cr, Mn, Cu, As, and Ni than other species. The heavy metal hierarchy in C. carpio was found to be Zn > Cu > Pb > Cr > Cd > Mn > As > Ni. While heavy metal levels in L. rohita and C. catla generally fell within reference ranges, exceptions were noted for Zn, Pb, and Cd. Conversely, in C. carpio, all metals exceeded reference ranges except for Cu and Ni. Principal Component Analysis (PCA) indicated a close relationship between water and sediment. Additionally, cluster analysis suggested that C. catla formed a distinct cluster from L. rohita and C. carpio, implying different responses to the environment. Despite concerns raised by the Geoaccumulation Index (Igeo) and Contamination Factor (CF), particularly for Cd, which exhibited a high CF. Furthermore, Hazard Index (HI) values for all three fish species were below 1, suggesting low health risks. However, elevated Igeo and CF values for Cd suggest significant pollution originating from anthropogenic sources. This study underscores the importance of monitoring heavy metals in water for both environmental preservation and human health protection. Future research efforts should prioritize pollution control measures to ensure ecosystem and public health safety., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The increasing levels of heavy metals in aquatic environments, driven by human activities, pose a critical threat to ecosystems' overall health and sustainability. This study investigates the bioaccumulation of heavy metals (Pb, Cu, Cr, and Cd) in water, sediment, and three fish species (Catla catla, Labeo rohita, Cirrhinus mrigala) of different feeding zones within Chashma Barrage, located in the Mianwali district of Punjab, Pakistan, on the Indus River. A comprehensive analysis, including an assessment of associated human health risks, was conducted. Thirty samples from all three sites for each fish species, with an average body weight of 160 ± 32 g, were collected from Chashma Barrage. Water quality parameters indicated suitability for fish growth and health. Heavy metal concentrations were determined using an atomic absorption spectrometer. Results indicated elevated levels of Cd, Cr, and Cu in sediment and Pb and Cd in water, surpassing WHO standard limits. Among the fish species, bottom feeder (C. mrigala) exhibited significantly (P < 0.05) higher heavy metal levels in its tissues (gills, liver, and muscle) compared to column feeder (L. rohita) and surface feeder (C. catla). Liver tissues across all species showed higher heavy metal bioaccumulation, followed by gills. Principal component analysis (PCA) revealed strong correlations among heavy metals in sediment, gills, muscle, and water in every fish species. However, the vector direction suggests that Cr was not correlated with other heavy metals in the system, indicating a different source. The human health risk analysis revealed lower EDI, THQ, and HI values (< 1) for the fish species, indicating no adverse health effects for the exposed population. The study emphasizes the bioaccumulation differences among fish species, underscoring the higher heavy metal concentrations in bottom feeder fish within Chashma Barrage., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: T helper interplay and cytokines monitoring in auto-immune skin disorders such as Pemphigus Foliaceus [PF] may play a central role in predicting the clinical stratification of the pathology., Objectives: In order to assess the CD4+ T cell imbalance, [i] this study aims to assess the related immune cells [Th1, Th2, Th17, and Treg cells] as well as the related cytokines [IL-1β, IFNγ, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-17A, IL-17F, IL- 22, TNF-β, and TNFα] in peripheral blood, and [ii] their respective transcription factors in the lesioned skin of PF endemic patients during the clinical course., Methods: Peripheral blood of 22 PF patients was analyzed by flow cytometry to assess the functional associations of Th cell subpopulations and their characteristic cytokines by multiplex bead assay of 14-plex cytokines. Skin mRNA expression of their associated transcription factors was analyzed using the TaqMan detection system., Results: Our findings revealed that the CD4+ T cell subtypes in PF patients compared to Healthy Controls [HC] were characterized by [i] a similar Th1/Th2 ratio and increased Th17/Treg ratio and [ii] significantly higher plasma levels of Th-17 specific cytokines; IL- 6, IL-8, IL-17A. Higher percentages in Th17 and Treg subtypes and a significant increase in plasma IL-17F levels were maintained in relapsing PF patients, arguing the pivotal role of Th17 cells in PF pathogenesis. Furthermore, our findings pointed out the major contribution of the pro-inflammatory cytokine IL-6. Indeed, in addition to being involved in the initial stages of disease development, IL-6 seems to also be involved in the maintenance of the pathophysiological process, probably through its effect on Th17 differentiation. The skin-relative mRNA expression levels of FOXP3 and TBET were significantly higher in relapsing PF patients compared to de novo PF patients., Conclusion: Our results highlight the central role played by Th17 lymphocytes and their related pro-inflammatory cytokines during the clinical course of the disease, reversing the Th1/Th2 dichotomy in PF., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Physicochemical properties of water influence planktonic diversity and distribution, which is essential in obtaining basic knowledge of aquatic biodiversity. Thus current study aims to investigate the spatiotemporal diversity, abundance ratio, and distribution of phytoplankton species and their association with water quality parameters of Chashma Lake, Pakistan. During the study period from 2018 to 2019, we measured 13 physicochemical parameters across three selected sampling sites (S1, S2, and S3) in Chashma Lake, revealing both spatial and temporal variability. Dissolved oxygen (DO) was higher in S3, while S1 exhibited higher alkalinity levels, carbon dioxide, phosphorus, and chloride levels. The study identified 77 phytoplankton species grouped into five taxonomic categories, with Cyanobacteria dominating (39.90%), followed by Chlorophyta (33.4%) and Bacillariophyta (24.88%). Euglenozoa and Ochrophyta were less abundant (1.3% and 0.41%, respectively). Spatial variations in phytoplankton distribution were noted, with Chlorophyta being more abundant at S2, Bacillariophyta and Cyanobacteria at S1, and Euglenozoa dominating at S3. Canonical Correspondence Analysis (CCA) revealed the influence of various physicochemical parameters on phytoplankton distribution. This comprehensive study provides valuable insights for the ecological assessment and monitoring of water bodies. It is recommended that continuous monitoring is required to capture long-term trends, further explore the specific environmental drivers impacting phytoplankton dynamics, and consider management strategies for maintaining water quality and biodiversity in Chashma Lake., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

This study investigates the influence of Withania somnifera root powder (WSRP) on different aspects of common carp ( Cyprinus carpio ), including growth, hematobiochemical parameters, antioxidant status, nonspecific immune response, and resilience to bacterial infections. Over a 60-day period, 180 common carp fingerlings (11.73 ± 0.52 g) were subjected to diets supplemented with increasing WSRP levels (0 (control), 1% (WSRP1), 2.5% (WSRP2.5), and 4% (WSRP4)). Nonspecific immune parameters were evaluated using serum samples collected at intervals of 0, 20, 40, and 60 days. After the feeding trial, the fish underwent experimental challenge with Aeromonas hydrophila , and relative percentage survival (RPS) was monitored for 14 days. The findings revealed a considerable ( p   < 0.05) enhancement in growth performance and a decreased feed conversion ratio (FCR) with increasing WSRP supplementation. Additionally, hematological and biochemical profiles exhibited improvements in groups receiving WSRP-enriched diets. Fish serum antioxidant status showed a significant ( p   < 0.05) increase, as indicated by increased activities of total antioxidant capacity (TAC), superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) in WSRP4. WSRP-supplemented diets led to remarkable enhancements in lysozyme activity ( p = 0.001), phagocytic activity ( p = 0.002), and RPS ( p = 0.003), peaking in WSRP4 at day 60. Furthermore, WSRP2.5 and WSRP4 demonstrated a substantial decrease ( p   > 0.05) in serum glucose and cortisol levels compared to other groups. In conclusion, WSRP proves valuable for enhancing growth, blood parameters, antioxidant balance, immune response, and infection resistance in common carp, especially at 2.5%-4% dietary supplementation levels. In the future, it will be crucial to study the long-term effects of WSRP supplementation on fish health, as well as its potential for large-scale aquaculture and economic viability in the aquafeed industry., Competing Interests: The authors declare there is no conflict of interest about this article., (Copyright © 2024 Syed Sikandar Habib et al.)

Introduction: This study investigates the potential effects of cannabis seed oil (CSO) on the wound healing process. The aim was to assess the efficacy of CSO in treating skin wounds using an animal model and to explore its anti-inflammatory properties through in silico analysis., Methods: Eighteen male albino Wistar rats, weighing between 200 and 250 g, were divided into three groups: an untreated negative control group, a group treated with the reference drug silver sulfadiazine (SSD) (0.01 g/mL), and a group treated topically with CSO (0.962 g/mL). The initial wound diameter for all groups was 1 cm. In silico studies were conducted using Maestro 11.5 to evaluate the anti-inflammatory effects of phytoconstituents against cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2)., Results: CSO and SSD treatments led to a significant reduction (p <0.05) in the size of burned skin wounds by day 5, with contraction rates of 53.95% and 45.94%, respectively, compared to the untreated negative control group. By day 15, wounds treated with CSO and SSD had nearly healed, showing contraction rates of 98.8% and 98.15%, respectively. By day 20, the wounds treated with CSO had fully healed (100%), while those treated with SSD had almost completely healed, with a contraction rate of 98.97%. Histological examination revealed granulated tissue, neo-blood vessels, fibroblasts, and collagen fibers in wounds treated with CSO. In silico studies identified arachidic acid, γ-linolenic acid, and linolenic acid as potent inhibitors of COX-1 and COX-2. Serum biochemical parameters indicated no significant changes (p > 0.05) in liver and kidney function in rats treated with CSO, whereas a significant increase (p < 0.01) in ALAT level was observed in rats treated with SSD., Discussion: The findings demonstrate that CSO has a promising effect on wound healing. The CSO treatment resulted in significant wound contraction and histological improvements, with no adverse effects on liver and kidney function.However, the study's limitations, including the small sample size and the need for detailed elucidation of CSO's mechanism of action, suggest that further research is necessary. Future studies should focus on exploring the molecular pathways and signaling processes involved in CSO's pharmacological effects., 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 Bouarfa, Chebaibi, Ez-Zahra Amrati, Souirti, Saghrouchni, El atki, Bekkouche, Mourabiti, Bari, Giesy, Mohany, Al-Rejaie, Aboul-Soud and Bousta.)

Controlling the hazard of sclerotia produced by the Sclerotinia sclerotiorum is very complex, and it is urgent to adopt an effective method that is harmonious environmentally to control the disease. Among the six isolates isolated from the rhizosphere of lettuce, the isolate HZA84 demonstrated a high activity in its antagonism towards Sclerotinia sclerotiorum in vitro, and produces siderophore. By amplification of internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF1-α), and RNA polymerase II subunit (RPB2) genes, the isolate HZA84 was identified as Trichoderma asperellum, which was confirmed by analysis of phylogenetic tree. The Scanning electron microscope monitoring detected that the isolate HZA84 spread over the sclerotial surface, thus, damaging, decomposing, and distorting the globular cells of the outer cortex of the sclerotia. The Real-time polymerase chain reaction (RT-qPCR) analysis disclosed the overexpression of two genes (chit33 and chit37) encoding the endochitinase in addition to one gene (prb1) encoding the proteinase during 4 and 8 days of the parasitism behavior of isolate HZA84 on the sclerotia surface. These enzymes aligned together in the sclerotia destruction by hyperparasitism. On the other hand, the pots trial revealed that spraying of isolate HZA84 reduced the drop disease symptoms of lettuce. The disease severity was decreased by 19.33 and the biocontrol efficiency was increased by 80.67% within the fourth week of inoculation. These findings magnify the unique role of Trichoderma in disrupting the development of plant diseases in sustainable ways., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Toxoplasma gondii is an apicomplexan protozoan parasite that infects one-third of the population of the world, including humans, animals, birds, and other vertebrates. The present investigation is the first molecular attempt in the Malakand Division of Pakistan to determine the epidemiology and phylogenetic study of Toxoplasma gondii infecting small ruminants., Methodology: A total of (N = 450) blood samples of sheep were randomly collected during the study period (December 2020 to November 2021), and DNA detection was done using PCR by amplifying ITS-1 genes. SPSS.20 and MEGA-11 software were used for statistical significance and phylogenetic analysis., Results: The overall prevalence of T. gondii infection among sheep was 14.44 % (65/450). A high infection rate was found in more than five-year-olds at 18.33 % (11/60). Sequencing and BLAST analysis of PCR-positive samples confirmed the presence of T. gondii. Randomly, three isolates were sequenced and submitted to GenBank under accession numbers (PP028089-PP028091), respectively. The BLAST analysis of the obtained sequences based on the ITS-1 gene showed 99 % similarities with reported genotypes found in goats of Malakand, Pakistan (PP028089) and dogs of Brazil (MF766454). The study concludes that T. gondii is notably prevalent among the sheep population in the region, emphasizing the significant role of risk factors in disease transmission across animals and potentially to humans. Further research, zoonotic potential analysis, and targeted control measures are warranted to address and manage this parasitic infection effectively., Competing Interests: Declaration of Competing Interest Authors confirm that they have no conflict of interest of any sort with any one., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Pesticides are frequently used to protect crop yields and manage malaria vectors; however, their inadvertent transport into aquatic habitats poses a significant concern. Various anthropogenic activities influence the Indus River in Pakistan. This study aimed to assess the presence of eight pesticide residues at three different sites (Kalabagh, Kundian, and Chashma) in water, sediment, and the fish species (Labeo rohita) during both dry and wet seasons to measure the intensity of this pressure. Pesticide analysis was carried out using gas chromatography equipped with an electron capture detector. The results revealed the highest concentrations of pesticides during both dry and wet seasons at all sites, measuring 0.83 and 0.62 μg/l (water), 12.37 and 9.20 μg/g/dw (sediment), and 14.27 and 11.29 μg/g/ww (L. rohita), respectively. Overall, pesticide concentrations were higher in the dry season than in the wet season across all study sites. Based on detection frequency and concentration in both seasons at all sites, dominant pesticides included cypermethrin and carbofuran (in water), as well as endosulfan and cypermethrin (in sediment and fish tissue). Levels of endosulfan and cypermethrin exceeded standard limits. Moreover, principal component analysis (PCA) indicated no correlation among pesticides in fish tissue, sediment, and water. However, pesticides exhibited different behavior in different seasons. Furthermore, endosulfan and triazophos impose great human health risk, as indicated by the THQ value (> 1). The overall HI value was greater for site 1 in the dry season (8.378). The study concluded that the presence of agricultural pesticides in the Indus River poses a risk to aquatic life and has the potential to disrupt the entire food chain. This highlights the importance of sustainable practices for the study area and Pakistan overall agricultural and environmental sustainability. It is further recommended to strengthen regulations for reduced pesticide use and promote eco-friendly pest management., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The ergosterol from mushrooms has gained significant ethnopharmacological importance in various cultures, including China, Japan, and Europe. This compound has been found to possess immune-boosting and anti-inflammatory properties, making it useful in the treatment of immune disorders. In this study, we focused on investigating the potential anticancer properties of ergosterol isolated from the edible mushroom Leucocalocybe mongolica in breast cancer cell lines. The ergosterol was purified and identified using advanced analytical techniques such as ESI-MS and NMR. We conducted cell proliferation assays on 4 T1 breast cancer cells to assess the cytotoxic effects of ergosterol. Furthermore, we analyzed the transcription levels of BAX, caspase-7, BCL-2, STAT-3, and PARP proteins using real-time PCR and Western blot analysis. Additionally, we employed non-targeted ultra-high-performance liquid chromatography and high-resolution mass spectrometry (UPLC-MS/MS) to study the potential mechanisms underlying the anticancer effects of ergosterol at the metabolomics level. The results demonstrated a significant reduction in cell viability and the induction of apoptosis upon treatment with ergosterol, especially at higher concentrations (P < 0.05). Moreover, ergosterol affected the expression of cancer-related genes, upregulating pro-apoptotic proteins such as BAX, caspase-7, and PARP, while downregulating the anti-apoptotic proteins BCL-2 and STAT-3 (P < 0.05). Western blot analysis confirmed these findings and provided further evidence of ergosterol's role in inducing apoptosis. Metabolomics analysis revealed substantial changes in pathways related to amino acid, antioxidant, and carbohydrate metabolism. In conclusion, our study demonstrates that ergosterol exhibits anticancer effects by inducing apoptosis and modulating metabolic pathways in breast cancer cells., 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 Author(s).)

The biosynthesis of silver nanoparticles (AgNPs) using plant extracts has become a safe replacement for conventional chemical synthesis methods to fight plant pathogens. In this study, the antifungal activity of biosynthesized AgNPs was evaluated both in vitro and under greenhouse conditions against root rot fungi of common beans ( Phaseolus vulgaris L.), including Macrophomina phaseolina , Pythium graminicola , Rhizoctonia solani , and Sclerotium rolfsii . Among the eleven biosynthesized AgNPs, those synthesized using Alhagi graecorum plant extract displayed the highest efficacy in suppressing those fungi. The findings showed that using AgNPs made with A. graecorum at a concentration of 100 μg/mL greatly slowed down the growth of mycelium for R. solani, P. graminicola , S. rolfsii , and M. phaseolina by 92.60%, 94.44%, 75.93%, and 79.63%, respectively. Additionally, the minimum inhibitory concentration (75 μg/mL) of AgNPs synthesized by A. graecorum was very effective against all of these fungi, lowering the pre-emergence damping-off, post-emergence damping-off, and disease percent and severity in vitro and greenhouse conditions. Additionally, the treatment with AgNPs led to increased root length, shoot length, fresh weight, dry weight, and vigor index of bean seedlings compared to the control group. The synthesis of nanoparticles using A. graecorum was confirmed using various physicochemical techniques, including UV spectroscopy, Fourier-transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) analysis. Collectively, the findings of this study highlight the potential of AgNPs as an effective and environmentally sustainable approach for controlling root rot fungi in beans.

The bioactive extracts of traditional medicinal plants are rich in polyphenols and help to rejuvenate skin. The study was designed to assess the skin rejuvenating effects of a stable cream enriched with 4% I. argentea (IaMe) extract. The quantity of polyphenols by spectrophotometric methods was TPC, 101.55 ± 0.03 mg GAE/g and total flavonoid content; 77.14 ± 0.13 mg QE/g, while HPLC-PDA revealed gallic acid; 4.91, chlorogenic acid 48.12, p-coumaric acid 0.43, and rutin 14.23 μg/g. The significant results of biological activities were observed as DPPH; 81.81% ± 0.05%, tyrosinase; 72% ± 0.23% compared to ascorbic acid (92.43% ± 0.03%), and kojic acid (78.80% ± 0.19%) respectively. Moreover, the promising sun protection effects Sun protection factor of extract (20.53) and formulation (10.59) were observed. The active cream formulation (w/o emulsion) was developed with liquid paraffin, beeswax, IaMe extract, and ABIL EM 90, which was stable for 90 days as shown by various stability parameters. The rheological results demonstrated the active formulation's non-Newtonian and pseudo-plastic characteristics and nearly spherical globules by SEM. The IaMe loaded cream was further investigated on human trial subjects for skin rejuvenating effects and visualized in 3D skin images. Herein, the results were significant compared to placebo. IaMe formulation causes a substantial drop in skin melanin from -1.70% (2 weeks) to -10.8% (12 weeks). Furthermore, it showed a significant increase in skin moisture and elasticity index from 7.7% to 39.15% and 2%-30%, respectively. According to the findings, Indigofera argentea extract has promising bioactivities and skin rejuvenating properties, rationalizing the traditional use and encouraging its exploitation for effective and economical cosmeceuticals., 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 Arshad, Shoaib Khan, Khan, Al-Roujayee, Mohany, Ahmad, Maryam, Akram, Shaukat, Khursheed and Aboul-Soud.)

Ralstonia solanacearum is a devastating soilborne pathogen which poses significant yield and economic losses to tobacco production globally. The impact of R. solanacearum on rhizosphere bacteriome and soil physicochemical characteristics in resistant and susceptible tobacco cultivars is poorly understood. This study aims to determine the effect of R. solanacearum on soil physicochemical parameters and rhizosphere bacteriome of resistant (K326) and susceptible (Hongda) tobacco cultivars at various growth stages. Results demonstrated that the contents of available potassium and phosphorus, as well as soil pH were significantly increased in K326 soils (CK and T2) compared with Hongda (T1) after 21, 42, and 63 days post-inoculation (dpi) of R. solanacearum except for available nitrogen which showed an opposite trend. The qPCR results showed a significant decrease in R. solanacearum population in rhizosphere of K326 (T2) compared to the Hongda (T1) at 21 and 63 dpi than that after 42 dpi. The rhizosphere bacteriome analysis through 16S rRNA amplicon sequencing revealed that rhizosphere bacterial community composition was significantly different between two tobacco cultivars (Hongda and K326) and this effect was more prominent after 63 dpi (93 days after post-transplantation), suggesting that each cultivar recruits a unique set of bacterial communities. There was no obvious difference observed in the rhizosphere bacteriome of CK (K326) and T2 (K326), which might be attributed to the same genetic makeup and inherent resistance of K326 to bacterial wilt infection. Analysis of co-occurrence networks revealed that the microbial network in T1 (Hongda) was more complex than those in T2 (K326) and CK (K326), while the networks in CK and T2 were almost identical. The present research highlights the time-course relationship between environmental factors and rhizosphere bacteriome of tobacco cultivars showing different levels of resistance against R. solanacearum. Conclusively, studying the plant-soil-microbe interaction system in susceptible and resistant tobacco cultivars may enable us to develop effective integrated disease control plans for the healthy production of tobacco crops., Competing Interests: Declaration of Competing Interest There is no financial and non-financial conflict of interest among authors., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Aims: Cognitive impairment poses a considerable health challenge in the context of type 2 diabetes mellitus (T2DM), emphasizing the need for effective interventions. This study delves into the therapeutic efficacy of quercetin, a natural flavonoid, in mitigating cognitive impairment induced by T2DM in murine models., Materials and Methods: Serum exosome samples were obtained from both T2DM-related and healthy mice for transcriptome sequencing, enabling the identification of differentially expressed mRNAs and long noncoding RNAs (lncRNAs). Subsequent experiments were conducted to ascertain the binding affinity between mmu-miR-129-5p, NEAT1 and BDNF. The structural characteristics and dimensions of isolated exosomes were scrutinized, and the expression levels of exosome-associated proteins were quantified. Primary mouse hippocampal neurons were cultured for in vitro validation, assessing the expression of pertinent genes as well as neuronal vitality, proliferation, and apoptosis capabilities. For in vivo validation, a T2DM mouse model was established, and quercetin treatment was administered. Changes in various parameters, cognitive ability, and the expression of insulin-related proteins, along with pivotal signaling pathways, were monitored., Key Findings: Analysis of serum exosomes from T2DM mice revealed dysregulation of NEAT1, mmu-miR-129-5p, and BDNF. In vitro investigations demonstrated that NEAT1 upregulated BDNF expression by inhibiting mmu-miR-129-5p. Overexpression of mmu-miR-129-5p or silencing NEAT1 resulted in the downregulation of insulin-related protein expression, enhanced apoptosis, and suppressed neuronal proliferation. In vivo studies validated that quercetin treatment significantly ameliorated T2DM-related cognitive impairment in mice., Significance: These findings suggest that quercetin holds promise in inhibiting hippocampal neuron apoptosis and improving T2DM-related cognitive impairment by modulating the NEAT1/miR-129-5p/BDNF pathway within serum exosomes., 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 Elsevier Inc. All rights reserved.)