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Hyalomma anatolicum is a tick of significant one-health importance due to its role as a vector for various pathogens affecting humans, animals and the environment, such as Theileria annulata, which causes tropical theileriosis in cattle, leading to severe economic losses. When infected with pathogens like T. annulata, the salivary glands of H. anatolicum undergo gene expression changes, secrete modified proteins and activate immune responses, all of which facilitate pathogen survival and transmission by modulating the host immune response and optimizing conditions for pathogen development. Understanding these responses is crucial for developing control strategies for tick-borne diseases. To understand the interaction between H. anatolicum and T. annulata, we performed a differential gene expression analysis of H. anatolicum salivary glands. An average of approximately 25 million raw sequencing reads were generated in each replicate using Illumina Sequencing. The sequenced reads were de novo assembled and the assembled transcriptome yielded approximately 50,231 non-redundant transcripts after clustering with CD-HIT using a sequence identity of 95% and alignment coverage of 90%. The assembly quality was evaluated with BUSCO analysis and found to be 86% complete using the Arachnida dataset and then blasted against non-redundant protein sequence database from NCBI followed by counting of reads and differential expression analysis. Overall, around 2400 and 400 genes were found differentially expressed with logFC > 1 and logFC > 2 respectively at FDR < 0.05. Top up-regulated genes included Calpain, Papilin, Neprilysin, and Ankyrin repeat-containing protein. Top down-regulated genes included Scoloptoxin, and Selenoprotein S and other uncharacterized proteins. Many other up-regulated proteins with high significance were uncharacterized suggesting room for further H. anatolicum functional and structural characterization studies. To our best knowledge, this is the first study of H. anatolicum sialotranscriptome which greatly contributes to sialotranscriptome information not only as sequence database but also indicates the potential targets for development of vaccine against ticks and transmission-blocking vaccines against T. annulata., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The use of wild food ingredients has been inextricably linked to each human culture; therefore, any cultural shift or transformation also mutates the knowledge. Particularly cross-cultural interactions have been playing a significant role in reshaping the knowledge within a given community. The study aimed to cross-culturally research the use of wild food plants among four different ethnolinguistic groups, i.e. Muhajir, Pathans, Punjabi, and Saraiki residing in the peri-urban area of Mianwali district, Punjab Pakistan. Data were taken through semi-structured interviews, and the results of cross-cultural comparison were visualized through Venn diagrams and statistically tested through Jaccard index. A total of 59 plants were recorded, which were used mostly as cooked vegetables (29 species) and snacks (21 species). About one-fourth of the quoted plants were commonly used by all the studied groups. High similarity on the use of the quoted plants was found among Muhajir, Punjabi, and Saraiki. Punjabi reported higher numbers of plants, and Pathans quoted comparatively lesser number. Pathans reported a comparatively high number of idiosyncratic foods uses followed by Muhajir. Punjabi, Muhajir, and Saraiki have a very close affinity and have comparatively rich knowledge after comparing those food uses which were reported by more 50% of participants. Punjabi frequently reported some plant uses which were rarely reported by the other four groups. Additionally a large number of plants were also quoted along with their medicinal uses which were prepared in the same way for both food and medicine. Despite the fact that most of the food ingredients were prepared in a simialr way but still the distinct names of several plants were retained across the studied groups. In the current context, the ethnobotanies of the studied communities are a blend as they might have learned the knowledge on the use of these plants from one another. It is concluded that since the knowledge is still in the memory of the people and has no serious threat to its extinction at hand, however it is necessary to frame policy programs in order to use this knowledge for the sustainability of future food and medical system otherwise it may be lost., Competing Interests: Declarations Ethics approval and consent to participate The research study proposal was approved by the Dean faculty of Sciences, Kohat University of Science and Technology, Pakistan, in 112 meeting of Advanced Studies Research Board (ASRB) held on 14 October 2023, under registration # B030211001. While conducting the field survey, the ethical guidelines as recommended by the International Society of Ethnobiology were precisely followed. All the participants provided prior oral consent before the interviews for data collection, photography, and sharing their knowledge and pictures with the public. Consent for publication All coauthors have read and agreed to submit the manuscript. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Women are the "guardians of the kitchen" and central to household food security, yet their role has never been studied across the Hindukush region or Pamiri Knot. This study explores the women's knowledge (specifically from the Khowar (Kho) and Wakhi linguistic groups) of local food systems and determines their role in ensuring household food security and sustainability in the mountain regions of northern Pakistan. Based on in-depth semi-structured interviews with female informants in the Rech and Broghil valleys of upper Chitral, 91 different types of food products were recorded, including wild and cultivated species. Eremurus stenophyllus and Allium barsczewskii were the commonly utilized plant species, though distinct preferences between the Kho and Wakhi groups were noted. Prominent differences were perceived in using certain cultivated plants among the two ethnic groups. For instance, Kho preferred plants like Beta vulgaris , Zea mays , and Brassica napus as indicated by the use reports, while Wakhi concentrated on Thymus serpyllum , Zygophyllum obliquum and Papaver involucratum. Both groups had shared dairy practices, but cottage cheese and curd were highly cited among Kho, while double-fermented curd and Qurut for Wakhi. The study recorded some new food uses for specific plants, such as Atriplex hortensis , Carthamus tinctorius , Hylotelephium spp., and Saxifraga sibirica . Cross-culture analyses revealed a mosaic pattern of homogenous and heterogenous trends based on reported food species of plants/animals and their use reports. Our findings emphasize the significant role of women in sustaining local food diversity, food sustainability, and the preservation and security of the local food systems, cultural legacy, and household food management. Therefore, inclusive research addressing their social, economic, and environmental issues must be conducted. Furthermore, policies must incorporate women's traditional knowledge to build resilient food systems.

Corosolic acid (CA), a pentacyclic triterpenoid, exhibits remarkably low hydrophilicity, restricting its application in aqueous systems. To enhance its hydrophilicity, we optimised nanoemulsion preparation conditions, resulting in a stable corosolic acid nanoemulsion system. By screening the oil phase, surfactant, and cosurfactant, along with investigating the mass ratio of surfactant and cosurfactant and the preparation temperature, we achieved an optimal corosolic acid nanoemulsion. We measured the particle size, polydispersity coefficient, and Zeta potential of the optimised formulation. The nanoemulsion's sustained-release effect, stability, and antibacterial activity were subsequently examined. The optimised formulation comprised ethyl oleate, cremophor EL, and Tween 80 (1.5:1), combined with ethanol in a ratio of 1:9:2.25 (w/w/w), and was prepared at 30 °C. This optimised corosolic acid nanoemulsion exhibited uniform particle size distribution, favourable dispersion, and notable slow-release capabilities. Importantly, the nanoemulsion demonstrated exceptional stability. In comparison to the positive control's bacteriostatic zone diameter, it was evident that the CA nanoemulsion (1.06 ± 0.11 mm) and blank nanoemulsion (1.03 ± 0.05 mm) both displayed notable inhibitory activity against S. aureus . Our findings established a solid foundation for the potential application of CA nanoemulsion in the food, cosmetics, and pharmaceutical industries. However, the application of CA nanoemulsion in real food or drug systems has not been explored yet., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Haimei Li reports financial support was provided by China Postdoctoral Science Foundation. Jingqi reports financial support was provided by 10.13039/100012547Guangxi Natural Science Foundation for Youth. Haimei Li reports financial support was provided by Guangzhou Postdoctoral Project Foundation. Lei zhang reports financial support was provided by Xinjiang research and development program project. Feng Hu reports financial support was provided by Yangxi Country Science and Technology Project. All authors declared that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

Background: This study was exclusively focused on the documentation and cross-cultural evaluation of ethnomedicinal knowledge (EMK) within the diverse linguistic groups of Kohistan situated between the Himalayan and Hindukush Mountain ranges in the north Pakistan., Methods: Data were gathered during the field survey (May 2022 to July 2023) through group conversations, semi-structured interviews, and on-site observation. Venn diagrams were employed to illustrate the comparative assessment of EMK, and different ethnobotanical indices were utilized to examine the data., Results: A total of 96 wild medicinal plant species (MPs) belonging to 74 genera and 52 botanical families were documented. The most reported MPs belong to the family Polygonaceae (11 species), followed by Asteraceae (9 species) and Lamiaceae (8 species). The ethnomedicinal uses of Leontopodium himalayanum, Pedicularis oederi, Plocama brevifolia, Polypodium sibiricum, Pteridium esculentum, Sambucus wightiana, Solanum cinereum, Teucrium royleanum, Rhodiola integrifolia, Aconitum chasmanthum were reported for the first time in this region. Among the reported taxa herbaceous species were dominated (72%), followed by trees and shrubs (17% and 10%, respectively). Digestive problems (40 taxa and 114 use reports) and skin disorders (19 taxa and 549 use reports) were the most cited disease categories, whereas M. communis, M. longifolia, Ajuga integrifolia, Ziziphus jujuba, and Clematis grata exhibited the highest percentage fidelity levels. Out of 109 documented medicinal uses, a mere 12 were shared across all linguistic groups, and Bateri emerges as a notable outlier with the highest number of medicinal uses. In addition, a significant homogeneity was noted in the reported botanical taxa (61 species) among different linguistic groups. However, since the last decade biocultural heritage of Kohistan is facing multifaceted risks that need urgent attention., Conclusion: Our findings could be valuable addition to the existing stock of ethnomedicinal knowledge and may provide ethnopharmacological basis to novel drug discovery for preexisting and emerging diseases prioritizing detailed phytochemical profiling and the evaluation of bioactive potential., (© 2024. The Author(s).)

Background: The use of medicinal plants to treat various veterinary illnesses has been practiced for millennia in many civilizations. Punjab is home to a diverse ethnic community, the majority of whom work in dairy farming, agriculture, and allied professions and have indigenous practices of treating animal illnesses using native flora. This study was designed to (1) document and preserve information about the applications of medicinal plant species in ethnoveterinary remedies among inhabitants of Punjab, Pakistan, and (2) identify popular plants for disease treatment by quantitative analysis of the obtained data and to assess the pharmacological relevance of these species., Methods: To collect data from informants (N = 279), questionnaires and semi-structured interviews were used. The ethnoveterinary data were analyzed using principal component analysis, relative frequency citation, fidelity level, relative popularity level, and rank order priority., Results: A total of 114 plant species utilized in the ethnoveterinary medicinal system were found, which were divided into 56 families and used to treat 16 different illnesses. The Poaceae family, with 16 species, was the most common in the region. The most commonly employed growth form in herbal preparation was herb (49%). The most used part in ethnoveterinary remedies was leaves (35%), while powder was the most commonly used way for preparing ethnoveterinary remedies (51 applications). According to principal component analysis, the most typically used species in the research region were grasses. Five grasses (Arundo donax, Desmostachya bipinnata, Eleusine indica, Hordeum vulgare, and Pennisetum glaucum) showed a 100% FL value when used to treat diuretics, helminthiasis, digestive problems, fever, cough, worm infestation, indigestion, galactagogue, oral infections, and genital prolapse. The maximum value of disease cured level (DCL%) was recorded at 87.6% for endo- and ecto-parasitic ailments in the study area., Conclusion: This study demonstrates that medicinal plants play an important part in satisfying farmers' animal healthcare demands, making it a feasible practice. The study also provides a wealth of knowledge regarding ethnoveterinary methods for further planning and application, providing an option for farmers who cannot afford allopathic therapy., (© 2024. The Author(s).)

Introduction: The utilization of fauna and fauna-based byproducts in ethnomedicinal usages has been a longstanding human activity, practiced across various cultures worldwide. This study focuses on investigating the utilization of animal-based traditional medicine by the people of Pakistan, specifically in the Gujranwala area., Methods: Data collection took place from January to September 2019 through interviews with local communities. Ethnomedicinal applications of animal products were analyzed using several indices, including Relative Frequency of Citation (RFC), Relative Popularity Level (RPL), Folk Use Value (FL), and Relative Occurrence Percentage (ROP)., Results: The study identified the use of different body parts of 54 species of animals in treating various diseases and health issues. These include but are not limited to skin infections, sexual problems, pain management (e.g., in the backbone and joints), eyesight issues, immunity enhancement, cold, weakness, burns, smallpox, wounds, poisoning, muscular pain, arthritis, diabetes, fever, epilepsy, allergies, asthma, herpes, ear pain, paralysis, cough, swelling, cancer, bronchitis, girls' maturity, and stomach-related problems. Certain species of fauna were noted by informers with high "frequency of citation" (FC), ranging from 1 to 77. For instance, the black cobra was the most frequently cited animal for eyesight issues (FC = 77), followed by the domestic rabbit for burn treatment (FC = 67), and the Indus Valley spiny-tailed ground lizard for sexual problems (FC = 66). Passer domesticus and Gallus gallus were noted to have the highest ROP value of 99., Discussion: The findings of this study provide valuable preliminary insights for the conservation of fauna in the Gujranwala region of Punjab, Pakistan. Additionally, screening these animals for medicinally active compounds could potentially lead to the development of novel animal-based medications, contributing to both traditional medicine preservation and modern pharmaceutical advancements., 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 Khan, Altaf, Hussain, Hamed, Safdar, Ayub, Memon, Hafiz, Ashraf, Amjad, Majeed, Hassan, Bussmann, Abbasi, Al-Yafrsi, Elansary and Mahmoud.)

Objective: To determine the genotypic frequency of Rh Cw antigen in blood donors of Northern Pakistan., Study Design: Descriptive cross-sectional study. Place and Duration of the Study: Department of Molecular Haematology, Armed Forces Institute of Transfusion (AFIT), Rawalpindi, Pakistan, from August 2022 to January 2023., Methodology: Blood donors were randomly selected. Venous blood samples were taken in K3-EDTA anticoagulant tubes. ABO and Rh D grouping were performed conventionally. DNA for Rh Cw genotyping was extracted via Chelex TM, followed by PCR amplification using an ABI 2700 thermal cycler. Human growth hormone (HGH) acts as an internal control. Amplified products underwent Polyacrylamide gel Electrophoresis (PAGE)., Results: There were 400 randomly chosen donors whose ages ranged from 26-35 years, with a predominantly male population (94.8%) of Punjabi origin (67.8%). The majority (87.3%) was RhD positive. Blood group B was the most prevalent (35%) in the studied population, followed by O (34.75%). Only 1.5% had Rh Cw antigen. Rh Cw was more prevalent in ABO-positive participants (87.25%) compared to ABO-negative (12.75%)., Conclusion: There was a 1.5% prevalence of Rh Cw antigen genotype in randomly selected Northern Pakistani blood donors. Rh Cw prevalence was higher in ABO-positive participants. Significant correlation (<0.05) existed between RhD and Cw antigens. Given the implications of anti-Cw antibody, including Cw antigen-positive cells in antibody screening is recommended., Key Words: Alloimmunisation, Blood donors, HDFN, Phenotype, Rh antigens, Transfusion.

Cedrus deodara is the central conifer plant affected by ozone and nitrogen pollutants among forest species worldwide. The growth of C. deodara depends upon the ectomycorrhizal (ECM) association, which is usually disturbed by these factors. This study aims to understand how these factors affect plants at physiological and biochemical levels. Three fungal strain consortiums were inoculated with two-year-old C. deodara seedlings. The stresses of 100 kg N h -1 and 100 ppb O 3 were applied for six months to study their impact on chlorophyll and antioxidant enzymes (SOD, CAT, and APX). The results showed that C2 (Consortium of Cedrus deodara ) positively impacted the growth of selected plant species. The high photosynthesis rate was determined by enhanced chlorophyll content, and C2-treated plants showed high chlorophyll content. Relatively, chlorophyll a and b contents increased significantly in the seedlings treated with Ethylenediurea (EDU) alone and with ozone stress. In addition, a significant difference was observed between EDU and O 3 -treated plants (14% EDU400-O 3 and 23% EDU600-O 3 ) and the control. Overall, antioxidant activities were higher in the treated samples than in the control. The order of SOD activity was C2 (448 U/gFW) and lowest (354.7 U/gFW) in control. APX also showed higher activity in treated plants in C1 ≥ C2 ≥ C3+O 3 , whereas CAT activity was the highest in C2 treatments. Ozone and nitrogen-stressed plants showed higher activities than EDU-treated plants compared to non-treated ones. Our findings highlight the importance of understanding the signaling effects of numerous precursors. Moreover, an extended investigation of seedlings developing into trees must be conducted to verify the potential of ectomycorrhizal strains associated with C. deodara and comprehend EDU's role as a direct molecular scavenger of reactive toxicants., Competing Interests: All authors declare that they have no conflict of interest., (© 2024 The Authors.)

This opinion piece, written by ethnobiologists from different parts of the world, emphasizes the importance of ethnobiology research in advancing contemporary biology, natural resource management, biodiversity conservation, sustainable development, and, especially, contributing to the ecological transition and more just and inclusive world. To achieve these goals, it is essential to develop research and collaborate with social groups that live in close relationship with nature in research activities, such as Indigenous Peoples and Local Communities (IPLC), as well as Afro-descendants and other Marginalized, Minority or Minoritized Communities (AMMC). Ethnobiology can identify and provide locally appropriate solutions to local problems, enabling sustainable resource management at the landscape level. The text explores important aspects that need to be considered to guide the future of ethnobiology in the next 20 years, aiming to integrate and amplify previous discussions held in the discipline and identify points that demand ongoing attention. This paper highlights reflections from diverse researchers, emphasizing how ethnobiology can embrace different perspectives and employ rigorous analysis of complex phenomena toward effective policies and practices. This approach holds the potential to address the challenges the planet is currently facing in the coming decades., (© 2024. The Author(s).)

Natural polysaccharides exhibit numerous beneficial properties, such as antioxidant, antitumor, hypoglycemic, and hypolipidemic activities. Moringa oleifera seeds are of high dietary and therapeutic value which drew a lot of attention. However, the regulation effect on anti-inflammatory activity of polysaccharides remains to be studied. Herein, novel bioactive polysaccharides (MOSP-1) were extracted from Moringa oleifera seeds, and the anti-inflammatory properties of MOSP-1 were uncovered. Ultrasound-assisted extraction (UAE) was used to prepare the polysaccharides with optimized conditions (70 °C, 43 min, and liquid-solid-ratio 15 mL/g). Then, DEAE-Sepharose Fast Flow columns were applied to isolate and purify MOSP-1. Rhamnose, arabinose, galactose, and glucose were identified as the monosaccharide constituents of MOSP-1, with a molecular weight of 5.697 kDa. Their proportion in molarity was 1:0.183:0.108:0.860 and 8 types of glycosidic linkages were discovered. Bioactive assays showed that MOSP-1 possessed scavenging activities against DPPH and ABTS radicals, confirming its potential antioxidation efficacy. In vitro experiments revealed that MOSP-1 could reduce the expression of inflammation-related cytokines, inhibit the activation of ERK, JNK, and p38 (the MAPK signaling pathway), and enhance phagocytic functions. This study indicates that polysaccharides (MOSP-1) from Moringa oleifera seeds with anti-inflammatory properties may be used for functional food and pharmaceutical product development., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Despite its millennial existence and empirical documentation, the ethnological knowledge of herbs is a more recent phenomenon. The knowledge of their historical uses as food, medicine, source of income and small-scale businesses, and the sociological impacts are threatened due to the slow ethnobotanical research drive. Species of the genus Solanum have long been extensively used in folk medicine to treat various illnesses of humans since the dawn of civilization. All data were systematically obtained from papers, monographs, and books written in Uzbek, Russian, and English through various scientific online databases, including Google, Google Scholar, PubMed, Scopus, Semantic Scholar, Science Direct, and Web of Science using specific keywords focused on eight Solanum species. Eight native and non-native Solanum species as S. dulcamara L., S. lycopersicum L., S. melongena L., S. nigrum L., S. rostratum Dunal., S. sisymbriifolium Lam., S. tuberosum L., and S. villosum Mill. have been recorded in Uzbekistan of Central Asia. In this article we presented recently obtained data on the diversity, morphological characteristics, global distribution, habitat, population status, phenology, reproduction, pharmacology and phytochemistry of these Solanum species in Uzbekistan. Furthermore, relying on a combination of literature reviews and analyses from various scientific papers, we focus on food consumption coupled with global ethnobotanical and ethnopharmacological uses in human diseases of the Solanum species growing in Uzbekistan. Since the dawn of civilization, these eight cultivated and non-cultivated species of Solanum have provided sustainable resources of medicinal plants in Uzbekistan to prevent and treat various human diseases. Based on the collected data, it was shown that Solanum species have not been studied ethnobotanically and ethnomedicinally in Uzbekistan and it is necessary to conduct phytochemical and biotechnological research on them in the future. Traditional uses and scientific evaluation of Solanum indicate that S. nigrum , S. sisymbriifolium and S. tuberosum are one of the most widely used species in some parts of the world. Although considerable progress has been made to comprehend the chemical and biological properties of S. nigrum and S. tuberosum species, more research on the pharmacology and toxicology of these species is needed to ensure the safety, efficacy, and quality of their biologically active extracts and isolated bioactive compounds. Additionally, conducting additional research on the structure-activity relationship of certain isolated phytochemicals has the potential to enhance their biological efficacy and advance the scientific utilization of traditional applications of Solanum taxa., 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 Gafforov, Rašeta, Zafar, Makhkamov, Yarasheva, Chen, Zhumagul, Wang, Ghosh, Abbasi, Yuldashev, Mamarakhimov, Alosaimi, Berdieva and Rapior.)

Salinity is known to have a greater impact on shoot growth than root growth. Na + buildup in plant tissue under salt stress has been proposed as one of the main issues that causes growth inhibition in crops via ionic imbalances, osmotic stress and pH disturbances. However, the evidence for apoplastic Na + buildup and the role of silicon in Na + accumulation at the subcellular level is still enigmatic. The current study focuses on the accumulation of Na + in the apoplast and symplast of younger and older leaves of two maize varieties (Iqbal as salt-tolerant and Jalal as salt-sensitive) using hydroponic culture along with silicon supplementation under short-term salinity stress. Subcellular ion analysis indicated that silicon nutrition decreased Na + concentration in both apoplastic washing fluid and symplastic fluid of maize under salt stress. The addition of silicon under NaCl treatment resulted in considerable improvement in fresh biomass, relative water content, chlorophyll content, and concentration of important subcellular ions ( i.e. , Ca 2+ , Mg 2+ , and K + ). Knowledge of subcellular ion analysis is essential for solving the mechanisms underlying vital cellular functions e.g. in the current study, the soluble Na + concentration in the apoplast of older leaves was found to be significantly greater (36.1 mM) in the salt-sensitive variety under NaCl treatment, which was 42.4% higher when compared to the Na + concentration in the salt-tolerant variety under the same treatment which can influence permeability of cell membrane, signal transduction pathways and provides insights into how ion compartmentalization can contributes to salt tolerance. Calcium silicate enrichment can contribute to increased growth and improved ionic homeostasis by minimizing leaf electrolyte leakage, improving mechanical functions of cell wall and reducing water loss, and improved photosynthetic function. In current investigation, increased water content and intracellular ionic homeostasis along with reduced concentration of Na + in the maize leaf apoplast suggest that calcium silicate can be used to ameliorate the adverse effects of salt stress and obtain yield using marginal saline lands., 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 Mahmood, Odeibat, Ahmad, Gatasheh, Shahzad and Abbasi.)

NAC transcription factors (TFs) are crucial to growth and defense responses in plants. Though NACs have been characterized for their role in several plants, comprehensive information regarding their role in Catharanthus roseus , a perennial ornamental plant, is lacking. Homology modelling was employed to identify and characterize NACs in C. roseus . In-vitro propagation of C. roseus plants was carried out using cell suspension and nodal culture and were elicited with two auxin-antagonists, 5-fluoro Indole Acetic Acid (5-F-IAA) and α-(phenyl ethyl-2-oxo)-Indole-Acetic-Acid (PEO-IAA) for the enhanced production of monoterpenoid indole alkaloids (MIAs) namely catharanthine, vindoline, and vinblastine. Analyses revealed the presence of 47 putative CrNAC genes in the C. roseus genome, primarily localized in the nucleus. Phylogenetic analysis categorized these CrNACs into eight clusters, demonstrating the highest synteny with corresponding genes in Camptotheca acuminata . Additionally, at least one defense or hormone-responsive cis-acting element was identified in the promoter region of all the putative CrNAC s. Of the two elicitors, 5-F-IAA was effective at 200 µM to elicit a 3.07-fold increase in catharanthine, 2.76-fold in vindoline, and 2.4-fold in vinblastine production in nodal culture. While a relatively lower increase in MIAs was recorded in suspension culture. Validation of RNA-Seq by qRT-PCR showed upregulated expression of stress-related genes ( CrNAC-07 and CrNAC-24 ), and downregulated expression of growth-related gene ( CrNAC-25 ) in elicited nodal culture of C. roseus . Additionally, the expression of genes involved in the biosynthesis of MIAs was significantly upregulated upon elicitation. The current study provides the first report on the role of CrNACs in regulating the biosynthesis of MIAs., 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 © 2023 Ahmed, Sajjad, Gatasheh, Ibrahim, Huzafa, Khan, Situ, Abbasi and Hassan.)

Industrial wastewater irrigation of agricultural crops can cause a lot of environmental and health problems in developing countries due to heavy metals deposition in agricultural soils as well as edible plant consumption by human beings. Therefore, this study was conducted to find out the heavy metals concentration in industrial wastewater and soil irrigated with that wastewater. In addition, the aim was to determine the impact of industrial wastewater irrigation on Parthenium hysterophorus and Zea mays genes involved in growth improvement and inhibition. For this purpose, plant samples from agriculture fields irrigated with wastewater from Hattar Industrial Estate (HIE) of Haripur, Pakistan, and control plants from non-contaminated soil irrigated with tape water were collected after 15 and 45 days of germination. Heavy metals concentration in the collected plant samples, wastewater, and soil was determined. The results revealed that the soil of the sample collection site was predominantly contaminated with Cr, Pb, Ni, Cu, Co, Zn, and Cd up to the concentrations of 38.98, 21.14, 46.01, 155.73, 12.50, 68.50, and 7.01 mg/kg, respectively. The concentrations of these heavy metals were found to surpass the permissible limit in normal agricultural soil. Expansins, cystatins (plant growth enhancers), and metacaspases (plant growth inhibitor) gene expression were studied through reverse transcription polymerase chain reaction. The results showed that the expression of these genes was higher in samples collected from wastewater-irrigated soils as compared to control. The expression of these genes was observed in 45 days old samples, 15 days old samples, and control. Taken together, this study suggests the use of Parthenium and maize for phytoremediation and that they should not be used for eating purposes if irrigated with industrial wastewater., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Hydatid cysts have been widely recognized for decades as a common medical problem that affects millions of people. A revolution in medical treatment may be on the prospect of nanotechnology enhancing chemotherapy against hydatid cysts. An overview of nanotechnology's impact on chemotherapeutics is presented in the current review. It discusses some of the challenges as well as some of the opportunities. The application of nanotechnology to enhance chemotherapy against hydatid cysts is what this review will explore. Nanotechnology is a critical component of delivering therapeutic agents with greater precision and efficiency and targeting hydatid cysts with better efficacy, and minimizing interference with surrounding tissue. However, there are biodistribution challenges, toxicity, and resistance problems associated with nanotherapeutics. Additionally, nanobiosensors are being investigated to enable the early diagnosis of hydatid cysts. A nanobiosensor can detect hydatid cysts by catching them early, non-invasively, rapidly, and accurately. The sensitivity and specificity of diagnostic tests can be enhanced with nanobiosensors because they take advantage of the unique properties of nanomaterials. By providing more precise and customized treatment options for hydatid cysts, nanotechnology may improve therapeutic options and strategies for diagnosing the disease. In conclusion, treatment with nanotechnology to treat hydatid cysts is potentially effective but presents many obstacles. Furthermore, nanobiosensors are being integrated into diagnostic techniques, as well as helping to diagnose patients earlier and more accurately.

This study aimed to address the pressing issue of plastic pollution in aquatic ecosystems by assessing the prevalence and distribution of microplastics (MPs) in water and riverbank sediments of the Pekalongan River, a vital water source in Indonesia. From the present findings, MP concentrations in water ranged from 45.2 to 99.1 particles/L, while sediment concentrations ranged from 0.77 to 1.01 particles/g. This study revealed that fragment and film MPs constituted 30.1 % and 25.4 % of the total, respectively, with MPs measuring <1 mm and constituting 51.4 % of the total. Colored MPs, particularly blue and black MPs, accounted for 34 % of the total. The primary polymer components, as determined via Fourier transform infrared spectroscopy, were identified as polystyrene, polyester, and polyamide. In response to the escalating plastic waste crisis caused by single-use plastics, Pekalongan's local government implemented refuse segregation and recycling programs as part of its efforts to transition toward zero-waste practices., 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 © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The main aim of this study was to assess the presence of microplastics in the water and sediments of the Surakarta city river basin in Indonesia. In order to accurately reflect the river basin, a deliberate selection process was employed to choose three separate sampling locations and twelve sampling points. The results of the study revealed that fragments and fibers were the primary types of microplastics seen in both water and sediment samples. Furthermore, a considerable percentage of microplastics, comprising 53.8 % of the total, had dimensions below 1 mm. Moreover, the prevailing hues identified in the water samples were blue and black, comprising 45.1 % of the overall composition. In contrast, same color categories accounted for 23.3 % of the microplastics found in the soil samples. The analysis of microplastic polymers was carried out utilizing ATR-FTIR spectroscopy, which yielded the identification of various types including polystyrene, silicone polymer, polyester, and polyamide., 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 © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Hemophagocytic lymphohistiocytosis (HLH) is a progressive and potentially life-threatening disorder. It is classified into primary and secondary HLH. The objective of our study was to determine the outcome of primary and secondary HLH in pediatric and adult patients based on HScore and treatment modality. We conducted a retrospective analysis done from July 2010 to June 2020. Variables analyzed included age, gender and history of death in siblings. HScore was used for disease classification while clinical and laboratory findings which were required to fulfill the HScore diagnostic criteria were also recorded. Continuous variables were summarized as median and categorical variables as frequencies and percentages. Categorical variables were compared using chi-square test and Fisher Exact test. Significance of different variables between primary and secondary HLH was calculated using independent-samples t test. A P value of < .05 was taken as significant. A total of 51 patients were included in the analysis (41 in primary and 10 in secondary HLH group). In primary HLH, 36 patients were in the pediatric age group and 12.2% had a history of death in sibling. All 41 patients had increased ferritin and decreased fibrinogen levels. The overall survival in primary HLH was 44%. In the secondary HLH group, viral infections were the most common etiology and ferritin was increased as well. The overall survival in secondary HLH was 60%. The median survival was 15 ± 4.8 months. The overall survival of both groups combined was 53%. Primary HLH should be considered in pediatric patients who present with pancytopenia and hepatosplenomegaly. In centers where genetic testing is not available, HScore along with serum ferritin and fibrinogen is a good substitute for disease classification., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2023 the Author(s). Published by Wolters Kluwer Health, Inc.)

Mushrooms play a crucial role in human life as well as in nature, providing food, medicine, and carrying out vital processes of decomposition, nutrient recycling, and developing mycorrhizal association with plants. The traditional system of knowledge about identification, collection, and usage of mushrooms has been accumulated through the shared experiences of many generations. Unfortunately, there have been continuous threats to the folk knowledge of mushrooms mainly due to habitat degradation, urbanization, and contemporary medication. The current research was, therefore, aimed to document an ethnomycological knowledge possessed by the ethnic communities of Swat, Pakistan. The purposive randomized sampling was carried out using chain referral method. Ethno-mycological information was collected from 62 informants using free listing, preference ranking, and use totaled methods. In total, 34 species of mushrooms belonging to 31 genera and 21 families were reported. About 85% of the reported species belong to Basidiomycetes, and 12.5% to Ascomycetes are used as food and for medicinal purposes. Morchella angusticeps , M. esculenta , Pleurotus sp., Auricularia sp., Flammulina velutipes , Agaricus bisporus , Ganoderma lucidum , and Sanghuangporus sanghuang were among the most cited edible and medicinal mushrooms. The current study revealed that district Swat is rich in wild edible and medicinal mushrooms (WEMs), and the local communities possess rich traditional knowledge about their collection, storage, and utilization. The diversity of WEMs of this region could contribute substantially to the socio-economic uplifting of the local communities through appropriate domestication and commercialization. Anthropogenic factors, coupled with depletion of traditional knowledge, threaten the diversity of WEMs in the region; therefore, in situ and ex situ conservation strategies are highly recommended.

Numerous investigations on plant ethnomedicinal applications have been conducted; however, knowledge about the medicinal use of wild animals is still limited. This present study is the second on the medicinal and cultural meaning of avian and mammalian species used by the population in the surrounding area of the Ayubia National Park, KPK, Pakistan. Interviews and meetings were compiled from the participants ( N = 182) of the study area. The relative frequency of citation, fidelity level, relative popularity level, and rank order priority indices were applied to analyze the information. Overall, 137 species of wild avian and mammalian species were documented. Of these, 18 avian and 14 mammalian species were utilized to treat different diseases. The present research showed noteworthy ethno-ornithological and ethno-mammalogical knowledge of local people and their connection with fauna, which might be useful in the sustainable utilization of the biological diversity of the Ayubia National Park, Khyber Pakhtunkhwa. Furthermore, in vivo and/or in vitro examination of the pharmacological activities of species with the highest fidelity level (FL%) as well as frequency of mention (FM) might be important for investigations on faunal-based new drugs.

Background: The mountainous territory of Kohistan shelters diverse food plant species and is considered one of the important hotspots of local plant knowledge. In the era of globalization and food commodification, wild food plants (WFPs) play an important role in supporting local food systems and related local knowledge is one of the important pillars of food sustainability across the region. Since the area is populated by different cultural groups and each culture has retained particular knowledge on the local plant species, therefore, to make a cross-culturally comparison, the study was planned to record and compare the local plants knowledge among three linguistic groups viz Gujjar, Kohistani and Shina in order to not only protect the local knowledge but to determine the food cultural adaptations among these groups looking through the lens of their food ethnobotanies., Methods: Field ethnobotanical survey was carried out in 2020-2021 to gather the data on wild food plants. We used semi-structured interviews. Use reports were counted, and the results were visualized through Venn diagrams., Results: In total, 64 plant species belonging to 45 botanical families were documented. Among these Ajuga integrifolia, Barbarea verna, Clematis grata, Impatiens edgeworthii, Ranunculus laetus (vegetables), Parrotiopsis jacquemontiana (fruit), Indigofera tinctoria (flower), Juniperus excelsa, Primula elliptica, P. macrophylla (flavoring agent), Leontopodium himalayanum (Chewing gum), and Juniperus excelsa (snuff) were reported for the first time. The highest use reports (≥ 90) were recorded for Mentha longifolia, Amaranthus hybridus, Quercus semecarpifolia, Solanum miniatum, Oxalis corniculata, Ficus palmata, and Urtica dioica. Maximum number of wild food plant species (WFPs) were reported by Kohistani, followed by Shinaki and Gujjari linguistic groups. The percentage overlap of traditional knowledge on WFPs was highest among Kohistani and Shinaki (56.0%), followed by Shinaki and Gujjars (17.0%), and Kohistani and Gujjars (15.0%). Kohistani and Shinaki groups exhibited maximum homogeneity in traditional knowledge. However, Gujjars had more knowledge on WFPs compared to Kohistani and Shinaki. In addition, some dairy products viz. Bhorus, Bagora, Bak, Cholam, Kacha, Gurloo and Poyeen were reported also reported that are consumed orally and used in traditional cuisines., Conclusions: The study indicates that Kohistan is one of the important spots of biocultural diversity and could be recognized as biocultural refugia. WFPs have been an integral part of the traditional food systems among the studied groups, particularly the Gujjars have reported more distinct plant uses which could be referred to their distinctive ecological experiences among others. However, social change is one of the challenges that might lead to the erosion of local plant knowledge. Moreover, intercultural negotiations among the studied groups are also a matter of concern which could homogenize the local knowledge among them. Therefore, we suggest solid policy measures to protect the local knowledge and celebrate diversity across this mountain territory., (© 2023. The Author(s).)

Land Surface Temperature (LST) affects exchange of energy between earth surface and atmosphere which is important for studying environmental changes. However, research on the relationship between LST, Land Use Land Cover (LULC), and Normalized Difference Vegetation Index (NDVI) with topographic elements in the lower Himalayan region has not been done. Therefore, the present study explored the relationship between LST and NDVI, and LULC types with topographic elements in the lower Himalayan region of Pakistan. The study area was divided into North-South, West-East, North-West to South-East and North-East to South-East directions using ArcMap 3D analysis. The current study used Landsat 8 (OLI/TIRS) data from May 2021 for LULC and LST analysis in the study area. The LST data was obtained from the thermal band of Landsat 8 (TIRS), while the LULC of the study areas was classified using the Maximum Likelihood Classification (MLC) method utilizing Landsat 8 (OLI) data. TIRS collects data for two narrow spectral bands (B10 and B11) with spectral wavelength of 10.6 μm-12.51 μm in the thermal region formerly covered by one wide spectral band (B6) on Landsat 4-7. With 12-bit data products, TIRS data is available in radiometric, geometric, and terrain-corrected file format. The effect of elevation on LST was assessed using LST and elevation data obtained from the USGS website. The LST across LULC types with sunny and shady slopes was analyzed to assess the influence of slope directions. The relationship of LST with elevation and NDVI was examined using correlation analysis. The results indicated that LST decreased from North-South and South-East, while increasing from North-East and South-West directions. The correlation coefficient between LST and elevation was negative, with an R-value of -0.51. The NDVI findings with elevation showed that NDVI increases with an increase in elevation. Zonal analysis of LST for different LULC types showed that built-up and bare soil had the highest mean LST, which was 35.76 °C and 28.08 °C, respectively, followed by agriculture, vegetation, and water bodies. The mean LST difference between sunny and shady slopes was 1.02 °C. The correlation between NDVI and LST was negative for all LULC types except the water body. This study findings can be used to ensure sustainable urban development and minimize urban heat island effects by providing effective guidelines for urban planners, policymakers, and respective authorities in the Lower Himalayan region. The current thermal remote sensing findings can be used to model energy fluxes and surface processes in the study area., Competing Interests: The authors declare no conflict of interest., (© 2023 The Author(s).)

Background: Fisheries have tremendous cultural and educational importance in human societies. The world is undergoing fast environmental and cultural changes, and local knowledge is being lost. Understanding how people interpret environmental change and develop practices in response to such change is essential to comprehend human resource use. This study was planned with the intent to document and conserve the knowledge about the uses of the freshwater fish fauna among the residents in South Punjab, Pakistan., Methods: Semi-structured interviews and questionnaires were conducted to collect data from informers (N = 88). Principal component analysis, relative frequency citation, fidelity level, relative popularity level, rank-order priority, and similarity index were used to analyze the fish data., Results: Overall, a total of 43 species of fishes were utilized in the study region, but only 26 species were utilized ethnomedicinally to treat a variety of illnesses such as asthma, body weakness, burn, chicken pox, cold, cough, eyesight, hepatitis, impotence, joint pain, night blindness, skin burn, spleen treatment, stomach infection, and weakness. The uses of fishes were analyzed employing various indices. The highest use value (UV) of 0.86 was calculated for spotted snakehead (Channa punctata), whereas the lowest UV of 0.05 was attained by karail fish (Securicula gora). Moreover, Channa punctata, Cyprinus carpio, Labeo rohita, Oreochromis niloticus, Wallago attu, Hypophthalmichthys molitrix, Rita rita, Sperata seenghala, Notopterus notopterus, Labeo dyocheilus, Systomus sarana, Puntius punjabensis, Securicula gora, Ompok bimaculatus, and Ompok pabda were the most popular species with RPL = 1.0. Out of the total, 20 species had a "zero" similarity index, while the ethnomedicinal use of 12 species (i.e., Labeo dyocheilus, Labeo boggut, Systomus sarana, Puntius punjabensis, Aspidoparia morar, Securicula gora, Crossocheilus diplochilus, Mastacembelus armatus, Ompok bimaculatus, Ompok pabda, Labeo gonius, and Sperata seenghala) was documented for the first time for a variety of diseases (i.e., body weakness, stomach infection, skin burn, joint pain, impotence, asthma, spleen treatment, and chicken pox)., Conclusion: Our findings showed that the local people of the study area hold noteworthy traditional knowledge about the medicinal and cultural uses of fish species. Furthermore, a comprehensive analysis of active chemicals and in vivo and/or in vitro activities of chemicals derived from ichthyofauna with the highest FC as well as UVs could be interesting for research on new drugs., (© 2023. The Author(s).)

In this paper, we propose a partial randomized response technique to collect reliable sensitive data for estimation of population proportion in ranked set sampling (RSS) scheme using auxiliary information. The idea is to increase confidence and (or) co-operation of the respondents by providing them the option of both 'direct' and 'randomized' response for the inquired sensitive question. This option is quite logical because perception of sensitive (insensitive) inquiry can vary among respondents. The properties of the proposed method are discussed and compared with existing randomized response techniques. Cost analysis is also carried out to prove supremacy of the suggested method. Finally, an application to clinical trial on AIDS is included., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Abbasi 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.)

The contamination of toxic trace metals in the food chain is one of the major threats to human health. Milk is part of a balanced diet, which is essential for proper growth, but the ingestion of contaminated milk may cause chronic health disorders. The present study is focused on the assessment of contamination of toxic trace metals in buffalo milk and the associated health risks to the consumers of Abbottabad, Pakistan. Standard analytical methods were employed to quantify the metal contents in the milk samples collected from various shops and homes in the months from June 2021 to October 2021. Health risk assessment was accomplished by computing estimated daily intake (EDI), health risk index (HRI), target hazard quotient (THQ), hazard index (HI), and target cancer risk (TCR). On a comparative basis, the mean concentration of Cr was found to be highest in both shop and home milk samples (101.3 ± 45.33 and 54.11 ± 24.20 mg/L, respectively), followed by Pb, Zn, Ni, and Cd levels. In buffalo milk collected from homes, the highest concentration of the metals was found in October, followed by July, September, June, and August. In shop milk, the increasing trend of metal contents was July > October > September > June > August. Significantly strong positive relationships were noted between the metal concentrations in the milk samples. Multivariate cluster analysis and principal component analysis exhibited significant anthropogenic contributions of the metals in buffalo milk. Mostly, the EDI and HRI values were exceeding the recommended limits; however, THQ, HI, and TCR showed that the intake of these metals through milk consumption was within the safe limit and thus revealed no significant carcinogenic or non-carcinogenic risks to the consumers. It is high time to ensure the continuous monitoring of organic/inorganic toxins in the milk and concerned authorities should take strict measures to control the contamination of milk and other food products.

The Moroccan flora abounds and is an important reserve of medicinal plants. Nigella sativa and Lepidium sativum are plants that are widely used in traditional medicine for their multiple therapeutic properties. The current study aims to highlight the biological activities that can justify and valorize the use of these plants. Flavonoids, total phenols, condensed tannins, and sugars were determined. The biological activities tested were antioxidant by determining the IC 50 (defined as the concentration of an antioxidant required to decrease the initial concentration by 50%; inversely related to the antioxidant capacity), hemagglutination, and hemolytic activities. Phytochemical quantification of the seed extracts indicated that the total phenol content was largely similar for both plants and in the order of 10 mg GAE (Gallic acid equivalent)/g. On the other hand, L. sativum seeds registered a higher content of flavonoids (3.09 ± 0.04 mg QE (quercetin equivalent)/g) as compared to Nigella saliva (0.258 ± 0.058). Concerning condensed tannins, N. saliva seeds present a higher amount with a value of 7.2 ± 0.025 mg/g as compared to L. sativum (1.4 ± 0.22 mg/g). Concerning the total sugar content, L. sativum shows a higher content (67.86 ± 0.87 mg/g) as compared to N. sativa (58.17 ± 0.42 mg/g); it is also richer in mucilage with a content of 240 mg as compared to 8.2 mg for N. saliva . Examination of the antioxidant activity using a DPPH (2.2-diphenyl 1-pycrilhydrazyl) test revealed that the EButOH (n-butanol extract) and EAE (ethyl acetate extract) extracts were the most active, with IC 50 values of 48.7 and 50.65 μg/mL for the N. sativa extracts and 15.7 and 52.64 μg/mL for the L. sativum extracts, respectively. The results of the hemagglutination activity of the different extracts of the two plants prepared in the PBS (phosphate-buffered saline) medium showed significant agglutination for the L. sativum extract (1/50) compared to the N. sativa extract (1/20). An evaluation of the hemolytic effect of the crude extract of the studied seeds on erythrocytes isolated from rat blood incubated in PBS buffer compared to the total hemolysis induced by distilled water showed a hemolysis rate of 54% for Nigella sativa and 34% for L. sativum . In conclusion, the two plants studied in the current work exhibited high antioxidant potential, which could explain their beneficial properties.

Background: Eastern Khyber Pakhtunkhwa is home to a vast range of medicinal and edible waterbird species due to its diverse geographical environment. Waterbird species have been used for various ailments and cultural practices since ancient times, while ethno-pharmacological applications and cultural uses of waterbird species in this area have seldom been documented. This study is the first ethnomedicinal and cultural assessment of waterbird species, and the first compilation and listing of all known data on these species in Eastern Khyber Pakhtunkhwa, Pakistan., Methods: Interviews and questionnaires were used to collect data from native respondents (N = 100). To analyze the data, principal component analysis (PCA), relative frequency of citation (RFC), fidelity level (FL%), relative popularity level (RPL), rank order priority, and similarity index were used., Results: In total, 64 waterbird species were utilized in cultural practices, of which 40 species are used to cure different infectious and chronic diseases such as cold, cough, flu, fever, respiratory disorders, asthma, TB, gastric ulcers, kidney stones, male impotency, obesity, paralysis, piles, cancer, arthritis, body pain, and weakness. PCA showed significant differences in the use of waterbird species among the local inhabitants of the study area, separated along the axis-2 (p < 0.05). The FL% of waterbird species varied from 12 to 100%. 100% FL was analyzed for four waterbird species, i.e., Charadrius mongolus (cold), Gallicrex cinerea (asthma), Anas platyrhynchos (cancer), and Esacus recurvirostris (body weakness). In this study, Mallard (Anas platyrhynchos) was the most popular species used in the healthcare system of Eastern Khyber Pakhtunkhwa, with high RFC (4.06), FL% (100), and RPL (1.0) values., Conclusion: We concluded that waterbird species are more used for medicine and food purposes in the study area. However, in vitro/in vivo assessment of biochemical activities of waterbird species with a maximum FL% might be significant to produce novel drugs. Recent research shows important ethno-ornithological information about native people and their links with waterbird species, which might be helpful for the sustainable use of waterbird diversity in the research area., (© 2022. The Author(s).)

Amphibians and reptiles have interacted with humans for millennia. However, humans interact with amphibian and reptile species in different manners, which depend on their culture and traditions. This study was designed to better understand the interactions between amphibian and reptile species and their usage among the native peoples in the vicinity of the Jhelum and Chenab rivers, Pakistan. Information was collected through semi-structured interviews and questionnaires, and was analyzed by using different indices, including the frequency of citation, corrected fidelity level, fidelity level, relative importance level, and informant major ailment. Two amphibians and twenty-six reptile species were used in therapeutic medicine in the study area. Based on the cultural analysis, we found that Naja naja (black cobra) was highly cited across all cultural groups. A 100% Fidelity Level was calculated for the following species: Naja naja (eye infection), Varanus bengalensis (joint pain), Eurylepis taeniolatus (cataract), and Acanthodactylus cantoris (cancer). We found five endangered species in the study area, i.e., Aspideretes gangeticus, A. hurum, Chitra indica, Varanus flavescens , and Geoclemys hamiltonii , that were used to cure joint pain, muscle stretching and pain, backbone pain, paralysis, and psoriasis, respectively. Likewise, Lissemys punctata andersoni , a vulnerable species as labelled by the International Union for Conservation of Nature, was extensively used for the treatment of joint pain, body pain, paralysis, and arthritis in the study area. In terms of conservation, it is critical to protect the highly vulnerable and endangered species that are being used in therapeutic medicines. Our findings may be helpful for the conservation of amphibian and reptile species by helping to make an effective plan to prevent their extinction. The main threats to the diversity of amphibian and reptile species in the area are hunting, trading, and cultural use. These threats could potentially lead to the extinction of these species. Therefore, with the involvement of concerned authorities, e.g., local stakeholders, the Ministry of Climate Change, provincial wildlife departments, academia, and conservation managers, immediate conservation measures should be taken for the protection and sustainable utilization of medicinal species.

Background: The use of animals and animal-derived products in ethnopharmacological applications is an ancient human practice that continues in many regions today. The local people of the Himalayan region harbor rich traditional knowledge used to treat a variety of human ailments. The present study was intended with the aim of examining animal-based traditional medicine utilized by the population of the Himalayan region of Azad Jammu and Kashmir. Methods: Data were collected from 2017 to 2019 through individual and group interviews. Data on traditional uses of animal products were analyzed, utilizing following indices such as the frequency of citation, use value, relative importance, similarity index, principal component analysis, and cluster analysis to find the highly preferred species in the area. Results: Ethnomedicinal uses of 62 species of vertebrates and invertebrates were documented. Flesh, fat, bone, whole body, milk, skin, egg, head, feathers, bile, blood, and honey were all used in these applications. The uses of 25 animals are reported here for the first time from the study area (mainly insects and birds, including iconic species like the kalij pheasant, Lophura leucomelanos ; Himalayan monal, L. impejanus ; and western tragopon, Tragopan melanocephalus ). The diversity and range of animal-based medicines utilized in these communities are indications of their strong connections with local ecosystems. Conclusion: Our results provide baseline data valuable for the conservation of vertebrate and invertebrate diversity in the region of Himalayan of Azad Jammu and Kashmir. It is possible that screening this fauna for medicinally active chemicals could contribute to the development of new animal-based drugs., 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 Faiz, Altaf, Umair, Almarry, Elbadawi and Abbasi.)

Inorganic pollutant, specifically heavy metals' contamination, is a significant matter of concern and is one of the key contributors in various health disorders including cancer. However, the interaction of heavy metals (HMs) with lung cancer has rarely been explored yet. Therefore, the present study was intended with the aim to identify the interactions of HMs with the target protein "epidermal growth factor receptor (EGFR)" of lung cancer and explore potential drug candidates, which could inhibit the active site of EGFR against HM exposure. The molecular operating environment (MOE) tool was used to study the interactions of HMs with EGFR protein. The drug-drug interaction (DDI) network approach was used to identify the potential drug candidates, which were further confirmed and compared with the commercial medicines/control group. Various compounds of twenty-three HMs were docked with EGFR protein. Out of which tinidazole, thallium bromodimethyl, and silver acetate (Sn, Ti, and Ag compounds) showed strong interactions with EGFR based on lowest-scoring values (-20.42, -7.86, and -7.74 kcal/mol, respectively). Among 1280 collected drug candidates, three synthetic compounds viz., ZINC00602803, ZINC00602685, and ZINC06718468 and three natural compounds (berberine chloride, transresveratrol, and ellagic acid) depicted strong binding capacity with EGFR. Specifically, the scoring value of ZINC00602803 (-30.99 kcal/mol) was even lowest than standard lung cancer drugs (afatinib, erlotinib, and gefitinib). Our findings revealed that both natural and synthetic compounds having strong associations with EGFR protein could be potential candidates to inhibit the interaction between HMs and lung cancer protein and can also be used as an alternative for the prevention and treatment of lung cancer. However, in vitro and in vivo studies should be conducted to validate the aforementioned natural and synthetic compounds., Competing Interests: All authors declared that they have no conflict of interest., (Copyright © 2022 Zainab Ayaz et al.)

Flubendiamide is a new class of chemical pesticide with broad spectrum activity against lepidopteran pests. Due to limited approach and high specificity towards various non targeted organisms, the unrestricted application of this pesticide as a prominent alternate for organochlorine and organophosphate pesticides, causing serious environmental pollution. In this study, wastewater was used for the determination of microbial strains and pesticide degrading fungi. Microbial population and flubendiamide resistant fungal strains were characterized using enriched medium. Aerobic bacteria (6.38 ± 0.23 log CFU/mL), nitrifying bacteria (2.73 ± 0.31 CFU/mL), Lactobaillus (0.72 ± 0.03 log CFU/mL), actinomycetes (5.36 ± 0.27 log CFU/mL) and fungi (4.79 ± 0.22 log CFU/mL) were detected. The prominent fungi genera were, Fusarium, Trichoderma, Cladophialophora, Paecilomyces, Talaromyces, Penicillium, Aspergillus, Candida, Phyllosticta, Mycosphaerella, Ochroconis, and Mucor. Minimum inhibitory concentration of the rapidly growing organism (FR04) revealed its ability to tolerate up to 1250 mg/L flubendiamide concentration. Morphological, biochemical and molecular analysis revealed that the strain was Aspergillus terreus FR04. The residual pesticide was detected using a High Performance Liquid Chromatography (HPLC). High performance liquid chromatography analysis revealed that 89 ± 1.9% pesticide removal efficiency was observed in strain FR04 at optimized culture conditions (96 h, pH 6.5, 30 °C and 300 mg/L pesticide concentration). The strain FR04 degraded pollutants from the wastewater and improved water quality. A. terreu sFR04 is an indigenous fungus and has the ability to degrade trizole pesticides from the wastewater significantly., (Copyright © 2021. Published by Elsevier Inc.)

Oxytetracycline is widely used in veterinary and human medicine. It has been detected in wastewater from pharmaceuticals, hospitals and domestic wastewater. In recent years, much more attention has been directed towards glutathione transferases (GSTs) because of their bio-transforming ability of antibiotics. In this study, 19 Lactobacillus strains were initially screened for the production of GSTs and five strains were selected for biotransformation of oxytetracycline. Among the strains, L. fermentum LA6 improved oxytetracyline degradation than other strains. It was subjected to optimize GST production and optimum growth was achieved after 24 h incubation at 32 ± 2 °C and 200 mg/L initial oxytetracycline concentration. The biocatalyst was immobilized and antibiotic degradation efficiency was analyzed. The immobilized culture of L. fermentum LA6 improved biodegradation of oxytetracycline in the wastewater. At 50 mg/L initial antibiotic concentration, 53.2 ± 2.8% oxytetracycline degradation was achieved, however, it improved at 200 mg/L antibiotic concentration in the culture medium (89.1 ± 4.3%) after 24 h. The chemical oxygen demand (COD) of the wastewater decreased significantly after treatment. At 200 mg/L oxytetracycline concentration, COD removal was considerably high (93.6 ± 5.3 mg/L) than 150 mg/L oxytetracycline concentration in the medium. Antibiotic removal efficiency in immobilized form revealed that this method is highly suitable for the removal of antibiotics from the wastewater., (Copyright © 2021. Published by Elsevier Inc.)

Genetic diversity and Agro-climatic conditions contribute significantly to the agronomic and morphological features of the food plant species, and their nutraceutical potential. The present study was intended to evaluate the impact of growing conditions on total phenolic and total flavonoid contents, and in vitro antioxidant potential in the bulbs and leaves of onion varieties planted under diverse environmental conditions. Standard analytical methods were used to quantify total phenolic content (TPC), total flavonoid content (TFC), and free radicals’ scavenging/antioxidant capacity. The impact of climatic and soil conditions was assessed using statistical tools. In general, onion varieties cultivated at three different locations viz. Kalar Kahar, Lahore and Swabi exhibited significant variations in TPC and TFC, and antioxidant activities. The bulbs and leaves of Mustang (V1) variety planted at Lahore and Swabi had significantly (p < 0.05), high levels of TPC (659.5 ± 6.59, and 631.1 ± 8.58 mg GAE/100 g, respectively). However, leaves of Red Orb (V2) and bulbs of Mustang (V1), and Golden Orb (V6), harvested from Kalar Kahar depicted the highest concentration of TFC (432.5 ± 10.3, 303.0 ± 6.67, and 303.0 ± 2.52 mg QE/100 g DW, respectively). Likewise, bulbs of V1 planted at Kalar Kahar, Lahore and Swabi exhibited maximum inhibition of DPPH, ABTS, and H2O2 radicals (79.01 ± 1.49, 65.38 ± 0.99, and 59.76 ± 0.90%, respectively). Golden Orb (V6) harvested from Lahore had the highest scavenging of OH radical (67.40 ± 0.09%). Likewise, bulbs of V1 variety planted at KalarKahar and Swabi had significant capacity to scavenge ferric ions (415.1 ± 10.6 mg GAE/100 g DW), and molybdate ions (213.7 ± 0.00 mg AAE/100 g DW). Conversely, leaves of Amazon (V8), planted at Lahore and Swabi depicted significant levels of DPPH, ABTS, H2O2 radical scavenging (90.69 ± 0.26, 63.55 ± 1.06, 51.86 ± 0.43%, respectively), and reduction of ferric ions (184.2 ± 6.75 mg GAE/100 g DW). V6 leaves harvested from Lahore and that of Super Sarhad (V3) from Swabi showed the highest inhibition of OH radical (61.21 ± 0.79%), and molybdate ions (623.6 ± 0.12 mg AAE/100 g DW), respectively. Pearson correlation and principal component analysis revealed strong relationships of climatic conditions, soil properties and elevation with TPC, TFC and free radicals’ scavenging potential in the bulbs and leaves of onion varieties. The variations in the total phenolic and flavonoid contents, and antioxidant potential of different varieties, and their associations with climatic and soil factors revealed the complexity of the growing conditions and genetic makeup that imposed significant impacts on the synthesis of secondary metabolites and nutraceutical potential of food and medicinal plant species.

Traditional diets exist in all cultures and geographic regions, and they often represent healthy eating options. Traditional culinary preparations have, however, often undergone profound change, even in the isolated Himalayan region. Therefore, we adapted methods to identify traditional plant foraging activities to better understand their significance in food system sustainability, as well as to promote innovative local gastronomies. Information on wild food and foraging practices was gathered from varied ethnic groups such as Kashmiri, Gujjars, Pahari, Dogra, Bakarwal, Balti, Beda and Brokpa through interviews ( n = 716) and group discussions ( n = 67) in four bio-geographic regions of the Jammu and Kashmir Himalayas (J&KH). The data were subjected to ordination techniques (Principal Component Analysis) via R software Ver. 4.0.0. We documented 209 food species, of which 73% were plants and 27% animals, used by the inhabitants of four bio-geographic regions of J&KH. The highest number of food plant species was recorded in Indian Kashmir, followed by Jammu, Azad Kashmir and Ladakh (81, 65, 60 and 27 species, respectively), and the maximum number of animal species was reported in Indian Kashmir, followed by Azad Kashmir, Ladakh and Jammu (33, 21, 19 and 17 species, respectively). The Azad Kashmir and Indian Kashmir regions showed greater similarity, whereas the least overlap was observed between Kashmir and Ladakh. The PCA showed considerable variation between different regions, and specific groups of species were more related to one specific region than others. The reported uses of Abies pindrow, Acacia modesta, Bergenia ciliata, Bergenia stracheyi and Juglans regia among plants, and Jynx torquilla, Streptopelia orientalis and Tadorna ferruginea among animals, are novel for the gastronomy of this part of the Himalayan region. We also recorded for the first time from this region seven unique food preparations of wild animals. This study documented extensive traditional knowledge on the usage of wild species, and is the first scientific description of wild food species and their vernacular names in the Western Himalayas, Jammu and Kashmir. Our findings can contribute significantly to combating food insecurity by revitalizing and reconsidering the rich bio-cultural food heritage around which local traditional communities have developed their food systems.

Melatonin is effective in modulating metabolism and regulating growth and development in many plants under biotic and abiotic stress. However, there is no systematic quantification of melatonin effects on maize growth, gas exchange, chlorophyll content, and the antioxidant defense system. A meta-analysis was conducted on thirty-two currently available published articles to evaluate the effect of stress types, study types, and maize varieties on response ratio (ln RR ++ ) of "melatonin" to "control (no melatonin)" on plant growth, enzyme activities, gas exchange parameters, and photosynthetic pigments. Our findings revealed that melatonin application overall increased plant height, leaf area, root length, fresh and dry root weight and shoot weight, superoxide dismutase (SOD), peroxide (POD), catalase (CAT), ascorbate peroxidase (APX), soluble sugar and protein, photosynthetic rate, stomatal conductance, transpiration rate, chlorophyll, and carotenoid in maize leaf under stress conditions. In contrast, melatonin application decreased the levels of hydrogen peroxide (H 2 O 2 ), superoxide anion (O 2 - ), malondialdehyde (MDA), and electrolyte leakage. The categorical meta-analysis demonstrated that melatonin application to chilling stress resulted in higher SOD activity followed by salt stress. Melatonin application to all stress types resulted in higher POD, CAT and APX activities, except Cd stress, which had no effect on POD and decreased CAT by 38% compared to control. Compared to control, melatonin resulted in lower reactive oxygen species (ROS) and electrolyte leakage under no stress, Cd, drought, salt, lead, heat, and chilling stress in all study types (pot, growth chamber, hydroponic, and field), except O 2 content which was not affected in pot and growth chamber studies. It was concluded that melatonin alleviates oxidative damage by improving stress tolerance, regulating the antioxidant defense system, and increasing leaf chlorophyll content compared to control.

Present study was conducted to investigate the adsorption and ultrasound-assisted adsorption potential of silver nanoparticles (AgNPs) and silver nanoparticles loaded on chitosan (AgCS composite) as nano-adsorbents for methylene blue (MB) removal. AgNPs were synthesized using leaf extract of Ligustrum lucidum, which were incorporated on the chitosan’s surface for modification. UV−Vis Spectroscopy, FTIR, XRD, SEM, and EDX techniques were used to confirm the synthesis and characterization of nanomaterials. Batch adsorption and sono-adsorption experiments for the removal of MB were executed under optimal conditions; for fitting the experimental equilibrium data, Langmuir and Freundlich’s isotherm models were adopted. In addition, the antimicrobial potential of the AgNPs and AgCS were examined against selected bacterial and fungal strains. UV−Vis spectroscopy confirmed AgNPs synthesis from the leaf extract of L. lucidum used as a reducer, which was spherical as exposed in the SEM analysis. The FTIR spectrum illustrated phytochemicals in the leaf extract of L. lucidum functioning as stabilizing agents around AgNPs and AgCS. Whereas, corresponding crystalline peaks of nanomaterial, including a signal peak at 3 keV indicating the presence of silver, were confirmed by XRD and EDX. The Langmuir model was chosen as an efficient model for adsorption and sono-adsorption, which exposed that under optimum conditions (pH = 6, dye initial concentration = 5 mg L−1, adsorbents dosage = 0.005 g, time = 120 min, US power 80 W), MB removal efficiency of AgNPs was >70%, using ultrasound-assisted adsorption compared to the non-sonicated adsorption. Furthermore, AgNPs exhibited promising antibacterial potential against Staphylococcus aureus with the maximum zone of inhibition (14.67 ± 0.47 mm). It was concluded that the green synthesis approach for the large-scale production of metallic nanoparticles is quite effective and can be recommended for efficient and cost-effective way to eradicate dyes, particularly from textile wastewater.

Eltrombopag has been used in ITP and found its use in AA armamentarium recently. We retrospectively analyzed 61 patients at a tertiary care center in Pakistan from January 2015 to January 2021. They included patients with severe AA who were refractory to at least one course of immunosuppressive therapy and persistent/chronic ITP who have received at least one previous treatment for ITP. Responses to Eltrombopag in our population were comparable to real-world experiences while tolerable hepatotoxicity and GI issues were notable. We found Eltrombopag to be a safe and efficacious agent for treating patients with ITP and AA., Competing Interests: The authors do not declare any relevant conflicts of interest., (© 2022 Published by Elsevier Ltd.)

Colorectal cancer (CRC) is currently one of the most frequent malignant neoplasms, ranking 3rd in incidence and 2nd in mortality both in the USA and across the world. The pathogenesis of CRC is a complex interaction between genetic susceptibility and environmental factors such as exposure to metals. Therefore, the present study was intended to assess the imbalances in the concentrations of selected essential/toxic elements (Pb, Cr, Fe, Zn, As, Cd, Cu, Se, Ni, and Hg) in the serum of newly diagnosed colorectal carcinoma patients (n = 165) in comparison with counterpart controls (n = 151) by atomic absorption spectrometry after wet-acid digestion method. Serum carcinoembryonic antigen (CEA) of the CRC patients was determined using immunoradiometric method. Body mass index (BMI) which is an established risk factor for CRC was also calculated for patients and healthy controls. Conversely, average Ni (2.721 μg/g), Cd (0.563 μg/g), As (0.539 μg/g), and Pb (1.273 μg/g) levels were significantly elevated in the serum of CRC patients compared to the healthy donors, while the average Se (7.052 μg/g), Fe (15.67 μg/g), Cu (2.033 μg/g), and Zn (8.059 μg/g) concentrations were elevated in controls. The correlation coefficients between the elements in the cancerous patients demonstrated significantly dissimilar communal relationships compared with the healthy subjects. Significant differences in the elemental levels were also showed for CRC types (primary colorectal lymphoma, gastrointestinal stromal tumor, and adenocarcinoma) and CRC stages (stage-I, stage-II, stage-III, and stage-IV) among the patients. Majority of the elements demonstrated perceptible disparities in their levels based on dietary, habitat, gender, and smoking habits of the malignant patients and healthy subjects. Multivariate methods revealed noticeably divergent apportionment among the toxic/essential elements in the cancerous patients than the healthy counterparts. Overall, the study showed significantly divergent distribution and associations of the essential and toxic elemental levels in the serum of the CRC patients in comparison with the healthy donors., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature.)