Local people in the Sudhnoti district of Pakistan share a rich practice of traditional medicine for the treatment of a variety of ailments. We selected nine plants from the Sudhnoti ethnopharmacological tradition used for the treatment of infectious and inflammatory disease. Our aim was to evaluate the in vitro anti-infective potential of extracts from these species against multidrug-resistant (MDR) ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Enterobacter species) pathogens. Plant specimens were collected in the Sudhnoti district of Pakistan and vouchers deposited in Pakistan and the USA. Dried bulk specimens were ground into a fine powder and extracted by aqueous decoction and maceration in ethanol. Extracts were assessed for growth inhibitory activity against ESKAPE pathogens and biofilm and quorum sensing activity was assessed in Staphylococcus aureus. Cytotoxicity to human cells was assessed via a lactate dehydrogenase assay of treated human keratinocytes (HaCaTs). Four ethanolic extracts (Zanthoxylum armatum, Adiantum capillus-venaris, Artemisia absinthium, and Martynia annua) inhibited the growth of MDR strains of ESKAPE pathogens (IC50: 256 μg mL−1). All extracts, with the exception of Pyrus pashia and M. annua, exhibited significant quorum quenching in a reporter strain for S. aureus agr I. The ethanolic extract of Z. armatum fruits (Extract 1290) inhibited quorum sensing (IC50 32–256 μg mL−1) in S. aureus reporter strains for agr I-III. The quorum quenching activity of extract 1290 was validated by detection of δ-toxin in the bacterial supernatant, with concentrations of 64–256 μg mL−1 sufficient to yield a significant drop in δ-toxin production. None of the extracts inhibited S. aureus biofilm formation at sub-inhibitory concentrations for growth. All extracts were well tolerated by human keratinocytes (LD50 ≥ 256 μg mL−1). Chemical analysis of extract 1290 by liquid chromatography-Fourier transform mass spectrometry (LC-FTMS) revealed the presence of 29 compounds, including eight with putative structural matches. In conclusion, five out of the nine selected anti-infective medicinal plants exhibited growth inhibitory activity against at least one MDR ESKAPE pathogen at concentrations not harmful to human keratinocytes. Furthermore, Z. armatum was identified as a source of quorum quenching natural products and further bioassay-guided fractionation of this species is merited.

Recently, an interest has surged in utilizing indigenous medicinal plants to treat infectious illnesses and extract bioactive substances, highlighting the need to analyze medicinal plants for phytochemicals and bioactivities. The present study was aimed to evaluate the impact of different solvent systems (aqueous, ethanol, and methanol) used for extraction on total phenolics, total flavonoids, antioxidant, and antibacterial activity of three medicinal plants of Azad Kashmir (Achillea millefolium, Bergenia ciliata, and Aloe vera). High phenolic content was found in methanol extracts of B. ciliata (27.48 ± 0.58 mg GAE/g dry weight), A. vera (25.61 ± 0.33 mg GAE/g dry weight), and A. millefolium (24.25 ± 0.67 mg GAE/g dry weight). High flavonoid content was obtained in the ethanol extract of A. millefolium (27.13 ± 0.64 mg QE/g dry weight), methanol extract of B. ciliata (17.44 ± 0.44 ± 0.44 mg QE/g dry weight), and the methanol extract of A. vera (14.68 ± 0.67 mg QE/g dry weight). Strong 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (DPPH) was obtained with a methanol extract of B. ciliata (IC50 = 60.27 ± 0.20 µg/mL). With a zone of inhibition and a minimum inhibitory concentration ranging from 10.00 ± 0.66 to 24.67 ± 1.21 mm and 78 to 625 µg/mL, respectively, all of the studied plants demonstrated notable antibacterial activity against Staphylococcus aureus and Escherichia coli. A. vera showed greater antibacterial activity as compared to other plants under study while methanolic extract showed greater antibacterial activity than ethanolic and aqueous extract. The findings of this research support the use of these medicinal plants to treat a variety of diseases.

Pseudomonas aeruginosa (P. aeruginosa), one of the dangerous multidrug resistance pathogens, orchestrates virulence factors production through quorum sensing (QS). Since the exploration of QS inhibitors, targeting virulence to circumvent bacterial pathogenesis without causing significant growth inhibition is a promising approach to treat P. aeruginosa infections. The present study has evaluated the anti-QS and anti-infective activity of epigallocatechin-3-gallate (EGCG), a bioactive ingredient of the traditional green tea, against P. aeruginosa. EGCG showed significant inhibitory effects on the development of biofilm, protease, elastase activity, swimming, and swarming motility, which was positively related to the production of C4-AHL. The expression of QS-related and QS-regulated virulence factors genes was also evaluated. Quantitative PCR analysis showed that EGCG significantly reduced the expression of las, rhl, and PQS genes and was highly correlated with the alterations of C4-AHL production. In-vivo experiments demonstrated that EGCG treatment reduced P. aeruginosa pathogenicity in Caenorhabditis elegans (C. elegans). EGCG increased the survival of C. elegans by 23.25%, 30.04%, and 36.35% in a dose-dependent manner. The findings of this study strongly suggest that EGCG could be a potential candidate for QS inhibition as an anti-virulence compound against bacterial infection.

Background This is the first comprehensive report on the traditional and novel uses of medicinal plants practiced by the indigenous communities of the Sudhnoti district of Azad Jammu and Kashmir (AJK), Pakistan. The area is rich in folklore and indigenous medicinal knowledge due to a unique tribal composition and socioeconomic conditions. This study aimed to document traditional knowledge of native plant use by the local communities, particularly those used for therapeutic purposes. Methods Field surveys were conducted from September 2015 to March 2017. Interviews with 125 local inhabitants of different tribes, age groups, genders, and occupations were conducted using structured and semi-structured questions along with group discussions. Data gathered on plant uses, local names, and modes of application of each plant species were organized in tables. Ethnobotanical indices such as use value (UV) and cultural significance index (CSI) were used to produce quantitative information on the plant use category, frequency, and cultural preference of species. Reports on therapeutic uses of medicinal plants were compared with previous studies. Results In all, 88 plant species from 45 families were reported, out of which 67 (77%) were used in ethnomedical applications. Asteraceae, Rosaceae, Fabaceae, and Lamiaceae were the dominant families. Berberis lycium was the most valued plant species, followed by Zanthoxylum armatum and Taraxacum officinale. Mentha arvensis had the highest cultural significance, followed by Mentha longifolia, Punica granatum, and Zanthoxylum armatum. Leaves were the most preferred plant parts in the preparation of medicine exclusively or mixed with other parts. The most frequently used process of crude preparation of medicinal plants was cooking. Oral intake was the predominant route of administration. Conclusions Our comparative analysis confirmed that most of the plants documented have uses that match those previously reported for the region and other parts of the world, with the exception of novel medicinal uses for 11 plant species, including Verbascum thapsus for earache, Elaeagnus umbellata for hepatitis, Achillea millefolium for oral care, Dicliptera roxburghiana to prevent sunstroke in cattle, Rumex hastatus for allergy antidote, Pyrus pashia for hepatitis, and Nerium oleander for diabetes.

In this study, we reported an easy, rapid, cost-effective and environmentally benign method for the fabrication of silver nanoparticles (Ag-NPs) using Olea ferruginea as reducing, capping and stabilizing agent. For this, an aqueous extract of leaf and bark of O. ferruginea was treated with 1 mM AgNO3, which reduces Ag ions to Ag-NPs by establishing reddish brown color. The synthesized Ag-NPs were spherical crystals, with a mean size of 23 and 17 nm for leaf- and bark-mediated Ag-NPs, respectively. Fourier transform infrared spectroscopy affirmed the role of leaf and bark extracts of O. ferruginea as reducing, capping and stabilizing agent. These biosynthesized Ag-NPs showed profound antibacterial activity against Gram-negative (Pseudomonas aeruginosa and Escherichia coli) and Gram-positive (Streptococcus pneumonia and Staphylococcus aureus) bacteria. The highest antibacterial activity was shown by bark Ag-NPs against S. aureus (14.00 mm), while leaf Ag-NPs showed higher activity against S. pneumonia (13.00 mm). Additionally, they produced effective antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) as compared to plant extracts and positive control. It was observed that the bark-mediated Ag-NPs had higher percentage (90%) of scavenging potential than the leaf-mediated Ag-NPs (78%). The significance of the current study is the synthesis of eco-friendly, easy and cost-effective Ag-NPs as biomedical products.

Objective To assess the state of knowledge, perception, preparedness and satisfaction level of health care personals (HCPs) in Azad Jammu & Kashmir (AJK) towards COVID-19. Study design It was cross sectional survey conducted by personal interviews. Methodology The study was conducted by interviewing 302 HCPs from 7 major district headquarter hospitals of AJK from May to September 2020. The questionnaire included 29 questions about demography, perception, training, preparedness, knowledge and awareness of HCPs about COVID-19. The data was encoded and analyzed statistically. Results Majority of the respondents (78.5%) believed that corona-virus is a serious health threat. However, 10.6% perceived it as a bio-weapon and 4.3% as not dangerous. About 10% of doctors were not considering it very dangerous compared to 1% of paramedics and 2.5% of the supporting staff ( P = 0.003 ). About 55% of the respondents showed confidence of the steps taken by government for prevention of COVID-19 while 45% had an opposite view. More than 70% of the participants had no formal training and the ratio of trained personals was significantly higher ( 0.018 ) in paramedical staff (33%) than doctors (15%). About 53% of the respondents were not satisfied with the provision of PPEs and 64% were not satisfied with their work and wages situation. Majority of the respondents (97%) were observed to follow the protocols for personal protection. Conclusion Majority of HCPs perceive covid-19 as a serious health concern. However, most of the HCPs are neither well trained for the current pandemic nor fully satisfied about personal protection, work load, and wages.

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Majority of the mountain dwelling communities living in the Himalayas rely on traditional herbal medicines for primary healthcare needs. Present study was conducted in fairy meadows and allied valleys in District Diamir, Gilgit Baltistan autonomous territory in northern Pakistan. Documentation of traditional medicinal knowledge (TMK) of local communities for the treatment of gastrointestinal disorders was carried out as a component of a wider medico-botanical expedition conducted in the entire base camp of the great Nanga Parbat peak during 2016–19. Various ethnobotanical parameters i.e. use value (UV), informant consensus factor (ICF), Fidelity level (FL), direct matrix ranking test (DMRT) and preference ranking (PR) were applied to evaluate the data collected during field surveys. The plants were also subjected to a comparative review for novelty assessment. A total of 61 medicinal plant species belonging to 55 genera and 35 families are reported here for the treatment of GIDs. Compositae was the leading family with 8 (13%) species. Fourteen gastrointestinal disorders were cured with 32% taxon were reported for stomachic followed by diarrhea (15%) and constipation (14%). Highest use reports (5) and use citations (207) were reported for Mentha longifolia L. while highest UV (1.79) was obtained for Artemisia maritima L. Hylotelephium telephioides (Ledeb.), A. maritima, M. longifolia, M. piperita L., Allium cepa L., and A. annua L. exhibited 100% FL. Highest ICF was calculated against dysentery and flatulence. DMRT ranked Prunus persica L. first for its multipurpose uses. Taking constipation as a reference gastrointestinal disease, PR for ten plant species was calculated where H. telephioides was ranked first followed by A. maritima. Present study concluded that 19 out of 61 plant species were documented for the first time with novel medicinal uses to cure GIDs. These plant species could act as potential reservoirs of novel lead compounds for the treatments of gastrointestinal disorders.