Ectomycorrhiza and ethylenediurea reduced the impact of high nitrogen and ozone stresses and increased the growth of Cedrus deodara.

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-1and 100 ppb O3 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 O3-treated plants (14% EDU400-O3 and 23% EDU600-O3) 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+O3, 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.

Advancing ethnobiology for the ecological transition and a more inclusive and just world: a comprehensive framework for the next 20 years.

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.

Ultrasonic extraction of Moringa oleifera seeds polysaccharides: Optimization, purification, and anti-inflammatory activities.

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.

Industrial wastewater irrigation increased higher heavy metals uptake and expansins, metacaspases, and cystatin genes expression in Parthenium and maize.

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.

A novel report on the occurrence of microplastics in Pekalongan River Estuary, Java Island, Indonesia.

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.

First evidence of microplastics in the water and sediment of Surakarta city river basin, Indonesia.

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.

Vernacular Taxonomy, Cultural and Ethnopharmacological Applications of Avian and Mammalian Species in the Vicinity of Ayubia National Park, Himalayan Region.

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.

Traditional Uses of Wild Edible Mushrooms among the Local Communities of Swat, Pakistan.

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.

Analysis of the relationship among land surface temperature (LST), land use land cover (LULC), and normalized difference vegetation index (NDVI) with topographic elements in the lower Himalayan region.

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.

Edible wild plant species used by different linguistic groups of Kohistan Upper Khyber Pakhtunkhwa (KP), Pakistan.

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.

Low apoplastic Na+ and intracellular ionic homeostasis confer salinity tolerance upon Ca2SiO4 chemigation in Zea mays L. under salt stress.

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., Ca2+, Mg2+, 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.

Genome-wide identification of NAC transcription factors and regulation of monoterpenoid indole alkaloid biosynthesis in Catharanthus roseus.

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 CrNACs. 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.

Exploring biodiversity and ethnobotanical significance of Solanum species in Uzbekistan: unveiling the cultural wealth and ethnopharmacological uses.

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.

Evaluation of Potentially Toxic Trace Metals and Associated Health Risk Assessment in Buffalo Milk.

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.

In Vitro Study of the Phytochemical Composition and Antioxidant, Immunostimulant, and Hemolytic Activities of Nigella sativa (Ranunculaceae) and Lepidium sativum Seeds.

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 IC50 (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 IC50 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.

Cultural and Medicinal Use of Amphibians and Reptiles by Indigenous People in Punjab, Pakistan with Comments on Conservation Implications for Herpetofauna.

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.

Traditional Uses of Animals in the Himalayan Region of Azad Jammu and Kashmir.

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.

In Silico Screening of Synthetic and Natural Compounds to Inhibit the Binding Capacity of Heavy Metal Compounds against EGFR Protein of Lung Cancer.

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.

Variations in Total Phenolic, Total Flavonoid Contents, and Free Radicals' Scavenging Potential of Onion Varieties Planted under Diverse Environmental Conditions.

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.

Melatonin Application Alleviates Stress-Induced Photosynthetic Inhibition and Oxidative Damage by Regulating Antioxidant Defense System of Maize: A Meta-Analysis.

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 (lnRR++) 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 (H2O2), superoxide anion (O2-), 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 O2 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.