Analysis of antidiabetic, antiulcer and analgesic potential of traditional ethnomedicinal plant Emex spinosa (L.) Campd. from Azad Jammu and Kashmir.

In this research antidiabetic, analgesic and antiulcer potential of traditional ethnomedicinal plant: Emex spinosa (L.) Campd. (Family Polygonaceae) was evaluated by extracting its phytoconstituents using methanol (MeOH) solvent through maceration protocol. The quantitative phytochemical analysis of the extract revealed flavonoids were highest in leaf extract (15.63±0.93 mg/mL) and with (11.5±0.57 mg/mL) in stem. Alkaloids and tanins were also present in the samples in various conc. while saponins were absent. To confirm pharmaceutical potential of plant against ulcer, diabetes and analgesic infirmities, a model experimental animal wistar albino rats (Rattus norvegicus) were used. In antiulcer study, using hot plate method the maximum results were observed with 250 mg/kg in the 2.5 hours of study. The leaf extract showed a 40.41±2.73 latency time and the fruit with a 36.77±2.41, and the stem with a 27.85±3.09, which was comparable to the standard drug Aspirin, i.e., 47.5±0.57. For analysis of antiulcer potential of the plants parts doses of 250 and 500 mg/kg was applied to check the reclamation of ethanol-induced acute ulcer and of Aspirin-induced chronic ulcer of stomach. In order to confirm efficacy of the drug potential of plant following parameters like microscopic evaluation, gastric volume, total acidity, mucosa weight, ulcer index, pH and histopathology of stomach were analyzed. In antidiabetic analysis, in an acute study after a single dose of 500 mg/kg extract after 2hrs the blood glucose levels were 367±51.09958NS, 416±59.79548NS, 437.5±61.96437NS mg/dL for leaf, stem and fruit, respectively. After4hrs 351.75±88.27644NS mg/dl, 448.25±25.64948NS mg/dl, 445.25±27.07205NS mg/dl and after 6hrs 354.5±92.70428NS, 442±24.60691NS, a440±33.16625NS mg/dl, respectively. The analgesic activity was explored by applying hot plate, tail flick and formalin paw licking method. In hot plate method the maximum results were observed with 250mg/kg in the 2.5 hours of study. The leaf extract showed a 40.41±2.73 latency time and the fruit with a 36.77±2.41 and the stem with a 27.85±3.09, which was comparable to the standard drug Aspirin, i.e., 47.5±0.57. The respective plant extracts at 250mg/kg showed a gradual rise in latency time when compared to the control. It was concluded that all three components of E. spinosa perform proved to be significant as potential source of herbal medicines to cure different prevalently occurring diseases. Furthermore, it was confirmed through results analysis that plant t can be used to discover novel drug using dedicated high throughput techniques and ethnopharmacological approaches.

Study of impacts of brickkiln emanations on soil quality of agriculture lands in selected areas of District Bhimber, Azad Jammu and Kashmir, Pakistan.

The pollution is hot issue of current era in world and the current study was carried to explore impacts of brickkilns' emanations on physiochemical properties of agricultural lands from District Bhimber of Azad Jammu and Kashmir (AJK) Pakistan. In this research, various edaphic characteristics: pH, soil organic matter, organic carbon, water holding capacity, cation exchange capacity and heavy metal contamination in soils nearby of brickkilns were determined. The pH of soil ranged from 5.55 to 7.50, soil organic matter was 0.35-0.90% and organic carbon content was 0.65-1.40%. The water holding capacity ranged from 2.10 to 3.20 mgL-1 and carbon exchange capacity was 1250 to 4202 meq/100g. The contamination profile of heavy metal depicted that Pb showed highest conc. 0.065 mg/g followed by Co (0.053 mg/g) and Ni with 0.52 mg/g in the soil. Pb and Cr had high conc. in soil samples around brickkilns due to burning of coal and rubber tyres as fuel. The conc. of sulphate and nitrate ranged from 0.90±0.50 mol L-1 to 4.25±0.65 mol L-1 and 2.30±0.50 mol L-1 to 6.55±0.25 mol L-1, respectively. The fertility of agriculture lands was depicted that edaphic properties were directly related while nutritive features were inversely commensurate to distance from brickkilns. The research proved that emanations of brickkilns causes severe impact on quality of agriculture land, plant growth and its yield. Hence, reclamation measures should be taken to mitigate and/or eradicate nuisance of brickkilns emanations by using environmental friendly strategies.

Ethnobotanical and conservation studies of tree flora of Shiwalik mountainous range of District Bhimber Azad Jammu and Kashmir, Pakistan.

The present study was carried out to explore and document traditional ethnobotanical knowledge of indigenous rural communities of Shiwalik mountainous range regarding tree flora of the area, District Bhimber of Azad Jammu and Kashmir, Pakistan. The local people of the area primarily depend on wild flora for life sustenance and cure of different infirmities. In this research, data was collected through visual appraisal and participatory rural approach using questionnaire method by applying semi-structured and structured-interview protocols (S4 Table). To validate and explore novelty of research work, various quantitative ethnobotanical indices like informant consensus factor, use value index, fidelity level, relative frequency of citation, relative importance of plants, rank order of priority, Jaccard index and priority ranking were used. The highest ICF was found for jaundice with 0.91 value followed by ICF values of 0.89, 0.86 and 0.85 for wounds, skin diseases and stomach pains, respectively. Fidelity level predicted that Azadirachta indica (Indian lilac tree) ranked first (100%) for its prevalent use in ethnomedicines. Highest use value index (0.61) was found for Azadirachta indica while the highest relative frequency of citation (55) was measured for Melia azedarach. The relative order of priority index was the highest for Azadirachta indica and Acacia arabica L. (ROP = 100) depicting their prevalent use in ethnomedicines. For EB study, the highest relative importance (50.8) was found for Azadirachta indica with good number of agro-uses and its seeds and bark are sold or marketed in local markets to prepare herbal therapeutics by indigenous practioners. The Jaccard index depicted that many traditional ethnomedicinal uses of prescribed trees were novel in recipe form or first time reported such as Senegalia modesta (Phulai) resin was used to prepare 'Panjoori' a local tonic prepared from cow or buffalo butter mixing with seeds of Papaver somnifera (Kashkhash) and Prunus amygdalus (Badaam). The wood of many trees like Pinus roxburghii (pine), Dalbergia sissoo (rosewood), Senegalia modesta (Phulai), Acacia. Arabica (kikar), Bombax cieba (cotton tree) is commercially sold as timber in markets (S3 Table). The research proved that population explosion and climate changes have triggered severe biotic and abiotic pressure on tree flora of the study area of Bhimber, AJK. The research describes that plants like Terminalia belerica (belerica), Terminalia arjuna (arjun tree), Cassia fistula (Indian laburnum), Butea monosperma (bastard teak), Phyllanthus emblica (Indian gooseberry), Morus laevigata (Shah toot), Bauhinia variegata (orchid tree) and Flacourtia indica (Indian plum/ kakoo) are threatened species and their population is highly reduced and if reclamation measures for their conservation are not taken, it may lead towards their complete loss from the area. This research recommends sustainable ethnobotanical use of tree flora, their growth and conservation for green and ecofriendly environment for safe and secure future of human generation.

Efficacy of Zn-Aspartate in comparison with ZnSO4 and L-Aspartate in amelioration of drought stress in maize by modulating antioxidant defence; osmolyte accumulation and photosynthetic attributes.

Human population is exceeding beyond the carrying capacity of earth resources and stresses like water shortage faced by the plants is jeopardizing the food security. Current research study was aimed to investigate the potentials of Zn-Aspartate (Zn-Asp), Zn-Sulphate (ZnSO4) and L-Aspartate (L-Asp) to be used as osmolytes and role of various levels of these chemicals in combating drought stress in maize plants in Punjab, Pakistan. Study was performed on two plots corresponding to drought and controlled environments. The lamina of maize plants was sprinkled row wise with various treatments including No spray (NS), water sprinkle (WS), sprinkle with ZnSO4 0.25% and 0.50%, sprinkle with Zn-Asp 0.25% and 0.50% and Foliar sprinkle of L-Asp 0.5% and 1%, respectively. Role of major osmoprotectants and secondary metabolites was analyzed and positive changes were found in total soluble sugars (41.16), flavonoids (5387.74), tocopherol content (9089.18), ascorbic acid (645.27) and anthocyanin (14.84) conc. which assists in mitigating drought menace on maize. Shoot mineral ions (Ca, K, Zn, P, Mg and N) status of water stressed maize plants was also analyzed and it was found that application experimental dose enhanced their availability to crop. Physio-biochemical studies were performed on antioxidants enzymes like superoxide dismutase (SOD), peroxidase (POD), carotenoid content (CC), malondialdehyde, hydrogen peroxide, aspartate and free amino acid contents. The activity of SOD was increased by 28.5% and activity of POD was increased by 33.33% due to foliar applied 0.5% Zn-Asp under drought stress. Photosynthetic pigments (chlorophyll A, B and total chlorophyll content) analysis was also carried out in this study. It was found that conc. of different chlorophylls pigments increased (chl-A: 2.24, chl-B: 25.12, total chl: 24.30) which enhanced photosynthetic activity culminating into better growth and yield). The level of malondialdehyde and hydrogen peroxide decreased by 43.9% and 32.8% respectively on treatment with 0.5% Zn-Asp proving the efficacy of the treatment in drought amelioration. Study reveals that Zn-Asp induced modulations are far better than conventional sulphate salts in mitigating water scarce environment. Current study recommends the use of the Zn-Asp to meet the global food and agricultural challenges as compared to ZnSO4 and L-Asp due to its better drought amelioration properties. This research provides valuable informations which can used for future research and practical use in agriculture fields by indigenous and other people to enhance yield of maize to meet the food necessities of country.

Inventorization of traditional ethnobotanical uses of wild plants of Dawarian and Ratti Gali areas of District Neelum, Azad Jammu and Kashmir Pakistan.

BACKGROUND: An ethnobotanical expedition was conducted to document the traditional ethnobotanical (TEB) uses of wild flora of Dawarian and Ratti Gali villages of District Neelam, Azad Jammu and Kashmir (AJK) Pakistan. District Neelam has rich plant diversity and is hub of many endemic plant species while the study areas are not yet explored. The research area: Dawarian and Rati Gali (DRG) area is mountaineous terrain and villages are located on far and farther distances. DRG area has rich biocultural and plant diversity comprising of different ethnic groups of Kashmir state. The current research was aimed to explore and document traditional medicines (TEMs) and other domestic and commercial uses of wild plants. This study will assist to evaluate conservation and commercial worth of wild flora which can be potential candidate for drug discovery through ethnopharmacological analysis. METHODS: The current quantitative ethnobotanical research was carried out in 2018 by interviewing 150 indigenous informants (90 male and 60 female) of DRG area using questionnaire applying structured and semi structured interview methodology. Data analysis was analyzed by using quantitative ethnobotanical statistical tools such as fidelity level (FL), informant consensus factor (ICF), Spearman's rank correlation (SRC) and data matrix ranking (DMR). RESULTS: The indigenous people of DRG area use wild plants in their daily life to cope life necessities i.e. food, vegetables, fodder, fuel, shelter, timber and herbal medicines. TEMs are primarily used to cure different infirmities like diabetics, asthma, dysentery, constipation, cold, fever, joint pain, wound healing, cancer, cardiovascular disorders, epilepsy, kidney infections and many types of skin diseases. Current study revealed the data of 103 wild plants species belonging to 46 plant families from selected areas of District Neelum, AJK. Results depicted that Asteraceae ranked 1st (12 plants spp). Among plant parts used leaf ranked 1st (18%), followed by seed (17%) and root (13%). While prevalent form recipe mode was decoction (20%), followed by powder (17%) and extract (14%) and fodder was highest (37%) EB use-form fodder, followed by food (32%) and fuel (17%). Quantitative ethnobotanical analysis (QEA) was carried to find the reliability and novelty of the study. Five plant species including Berberis lyceum (FL = 97.78%), Isodon rugosus (FL = 95.71%), Saussurea lappa (FL = 94.74%), Aconitum heterophyllum (FL = 92.71%) and Taxus baccata (91.58%) had shown high fidelity level which confirmed that these plants have high medicinal worth in study area. The highest value (0.94) of ICF was for diseases group "tuberculosis and leucorrhea", followed by stomachache and flatulence (0.93), diabetics and blood pressure (0.92) and asthma and chest infections (0.88). For other uses fuel with ICF (0.83) ranked first and second was hedging and thatching (ICF = 0.82) where people use plants or their parts for construction. Spearman's rank correlation (SRC) test indicated that number of TEB uses increases if number of species is increased. Jaccard index (JI) analysis depicted that 56.31% plants are being used as TEMs which are first time explored from the study area. While 26.21% plants are being used in different TEB uses which are different from past cited literature. These novel findings of research indicate that wild flora of the study area has great potential for novel drug discovery and provision of materialist services for the indigenous communities. CONCLUSION: The present research revealed that TEMs uses of 58 plants are novel being first time reported from the study area (DRG) of District Neelam of AJK. The results showed that plants like Acer cappadocicum, Ajuga bracteosa and Swertia paniculata are used to cure diabetes, Viscum album, Viola canescens, Taxus baccata are used for cure of cancer, Isodon rugosus, Polygala chinensis are used in TEMs for treating cardiovascular disorders and Anaphalis triplinervis is used for epilepsy. Berberis lyceum, Ajuga bracteosa, Aconitum heterophyllum, Bistorta amplexicaule, Saussurea lapa and Jurinea dolomiaea are severely threatened and there is urgent need to do conservation measures for available of valuable MPs to the indigenous communities for life necessities and for future research. The current study will also be useful addition in ethnobotanical database, preservation of traditional culture and drug discovery and drug development through future ethnopharmacological research.

Gathered Wild Food Plants among Diverse Religious Groups in Jhelum District, Punjab, Pakistan.

Recent ethnobotanical studies have raised the hypothesis that religious affiliation can, in certain circumstances, influence the evolution of the use of wild food plants, given that it shapes kinship relations and vertical transmission of traditional/local environmental knowledge. The local population living in Jhelum District, Punjab, Pakistan comprises very diverse religious and linguistic groups. A field study about the uses of wild food plants was conducted in the district. This field survey included 120 semi-structured interviews in 27 villages, focusing on six religious groups (Sunni and Shia Muslims, Christians, Hindus, Sikhs, and Ahmadis). We documented a total of 77 wild food plants and one mushroom species which were used by the local population mainly as cooked vegetables and raw snacks. The cross-religious comparison among six groups showed a high homogeneity of use among two Muslim groups (Shias and Sunnis), while the other four religious groups showed less extensive, yet diverse uses, staying within the variety of taxa used by Islamic groups. No specific plant cultural markers (i.e., plants gathered only by one community) could be identified, although there were a limited number of group-specific uses of the shared plants. Moreover, the field study showed erosion of the knowledge among the non-Muslim groups, which were more engaged in urban occupations and possibly underwent stronger cultural adaption to a modern lifestyle. The recorded traditional knowledge could be used to guide future development programs aimed at fostering food security and the valorization of the local bio-cultural heritage.

Ethno-veterinary uses of Poaceae in Punjab, Pakistan.

Plant species of the Poaceae family are not only used as fodder and forage but also contribute substantially to the treatment of various health disorders, particularly in livestock. Consequently, the present study was aimed to document the therapeutic uses of Poaceae practiced by the inhabitants of the Punjab Province for the treatment of various veterinary health disorders. Semi structured interviews, group discussion and field walks were conducted to collect the data. Quantitative indices including cultural significance index (CSI), relative frequency of citations (RFC), fidelity level (FL), relative popularity level (RPL), and Jaccard Index (JI) were used for the data analysis. Traditional uses of 149 species belonging to 60 genera and 16 tribes of 5 sub families of Poaceae were recorded. Whole plants and leaves were the most consistently used parts with 40.94 and 29.53%. The plants were mainly given orally as fodder (59 reports) without processing followed by decoction (35 reports). Most of the species were employed to treat infectious diseases (25.93%), and digestive disorders (14.10%). Triticum aestivum had the highest CSI, RFC and RPL levels at 8.00, 0.96, 1.00, respectively, followed by Oryza sativa and Poa annua. Likewise, T. aestivum and Saccharum spontaneum had 100% FL and ROP. Jaccard index ranged from 12.25 to 0.37. Twelve plant species namely Chrysopogon zizanioides (anti-inflammatory), Pennisetum lanatum (improve bull fertility), Cymbopogon citratus (glandular secretion), Sorghum saccharatum and Themeda triandra (malaria), Aristida funiculate (anticancer), Koeleria argentia (skin allergies), Tetrapogon villosus (antibacterial), Cynodon radiatus (eyes infection), Sporobolus nervosa (Jaundice), Enneapogon persicus (antifungal), and Panicum repens (dysfunctional cattle organs) were reported for the first time, with novel ethnoveterinary uses. The inhabitants of the study area had a strong association with their surrounding plant diversity and possessed significant knowledge on therapeutic uses of Poaceae to treat various health disorders in animals. Plant species with maximum cultural and medicinal values could be a potential source of novel drugs to cure health disorders in animals and human as well.

Chitosan-immobilized pectinolytics with novel catalytic features and fruit juice clarification potentialities.

Biological macromolecules are primarily composed of complex polysaccharides that strengthen microbial growth for the production of industrially relevant enzymes. The presence of polysaccharides in the form of the disrupted cell wall and cell materials are among major challenges in the fruit juice industry. The breakdown of such biological macromolecules including cellulose and pectin is vital for the juices processing. In this background, pectinolytic enzymes including polygalacturonase (PG), pectin lyase (PL), and pectin methylesterase (PME) were isolated from Aspergillus ornatus, statistically optimized and purified via ammonium sulfate fractionation (ASF), dialysis, and Sephadex G-100 gel permeation chromatography. After passing through Sephadex G-100 column, PG, PL, and PME were 2.60-fold, 3.30-fold, and 4.52-fold purified with specific activities of 475.2U/mg, 557.1U/mg, and 205.7U/mg. The active PG, PL, and PME, each separately, were surface immobilized using various concentrations of chitosan and dextran polyaldehyde as a macromolecular crosslinking agent. Prior to exploit for juice clarification purposes, various parameters including pH, thermal and Michaelis-Menten kinetic constants of purified and chitosan-immobilized fractions were investigated. A considerable improvement in the pH and thermal profiles was recorded after immobilization. However, the negligible difference between the Km and Vmax values of purified free and chitosan-immobilized fractions revealed that the conformational flexibility of pectinolytics was retained as such. A significant color and turbidity reductions were recorded after 60min treatment with CTS-PG, followed by CTS-PME, and CTS-PL. It can be concluded that the clarification of apples, mango, peach, and apricot juices was greatly affected by CTS-PG, CTS-PME, and CTS-PL treatments rendering them as potential candidatures for food industry applications.

Overexpression of a cotton cyclophilin gene (GhCyp1) in transgenic tobacco plants confers dual tolerance to salt stress and Pseudomonas syringae pv. tabaci infection.

The full-length cDNA of a cyclophilin-like gene was cloned from Gossypium hirsutum using rapid amplification of cDNA ends and was designated as GhCyp1, a member of the immunophilin protein family. GhCyp1 expression level was higher in roots and stems than in other tissues of cotton, as determined by real-time reverse transcription polymerase chain reaction (RT-PCR). To characterize the GhCyp1 gene, tobacco (Nicotiana tabacum) was transformed via Agrobacterium tumefaciens with a vector to express the gene under the control of a strong constitutive promoter, CaMV35S (Cauliflower Mosaic Virus). Based on analyses of tolerance to salinity stress and Pseudomonas syringae pv. tabaci (Pst) infection, the overexpression of GhCyp1 in transgenic plants conferred higher tolerance to salt stress and Pst infection compared with control plants. Therefore, we suggest that GhCyp1 may be a suitable candidate gene to produce transgenic plants with tolerance to abiotic and biotic stresses.