Pharmacological activities and gas chromatography-mass spectrometry analysis for the identification of bioactive compounds from Justicia adhatoda L.

The current study aimed to assess the pharmacological potential of Justicia adhatoda by evaluating the presence of biologically active compounds using the gas chromatography-mass spectrometry approach and to undertake biological activities for the effectiveness of the present compounds using standard tests. A total of 21 compounds were identified in the gas chromatography-mass spectrometry analysis of the ethyl acetate fraction in which 14 of the identified compounds are recognized for their pharmacological potential in the literature. In total, four fractions (ethyl acetate, chloroform, n-hexane, and aqueous) were evaluated for pharmacological activities. In carrageenan-induced inflammation, the chloroform fraction exhibited high anti-inflammatory activity (46.51%). Similarly, the analgesic potential of ethyl acetate fraction was the most effective (300 mg/kg) in the acetic acid-induced test. Similarly, in the formalin test, ethyl acetate fraction exhibited maximum inhibition in both early (74.35%) and late phases (88.38). Maximum inhibition of pyrexia (77.98%) was recorded for the ethyl acetate fraction (300 mg/kg). In DPPH assay, the ethyl acetate fraction revealed the highest scavenging potential among other fractions (50 µg/ml resulted in 50.40% and 100 µg/ml resulted in 66.74% scavenging).

Ethnogynaecological Knowledge of Traditional Medicinal Plants Used by the Indigenous Communities of North Waziristan, Pakistan.

Background: Since the beginning of civilization, medicinal plants have been used in human healthcare systems. Studies have been conducted worldwide to evaluate their efficacy, and some of the results have triggered the development of plant-based medications. Rural women in Pakistan frequently experience gynaecological disorders due to malnutrition and heavy physical work during pregnancy. Due to the low economic status, the remoteness of the area, and the lack of modern health services, herbal therapy for gynaecological disorders is common among the indigenous tribes of the study area. Methods: Field surveys were carried out from April 2018 to October 2020 to collect data regarding medicinal plants used for different gynaecological disorders. A semistructured questionnaire was used to collect ethnogynaecological data. Results: In total, 67 medicinal plant species belonging to 38 families are being used to treat 26 different gynaecological problems. The herbaceous growth form and the Lamiaceae family were recorded with the maximum number of plant species (42 species and 7 species, respectively). Leaves are the most highly utilized plant part, with 16 species. In the case preparation method, decoction was the dominant method (25 species, 36.76%). The informants reported the maximum number of species for the treatment of irregular menstrual flow as 11 species (15.28%). The highest relative frequency of citation (RFC) value was obtained for Acacia modesta (0.37), and the use value (UV) for Tecomella undulata (0.85). The highest informants' consensus factor (ICF) value (1.0) was obtained for emmenagogue and tonic each after delivery. The highest consensus index (CI%) value was calculated for Acacia modesta (36.92%). The Lamiaceae had the highest family importance value (FIV) (98.46%). Conclusion: This is the first ever quantitative study focusing mainly on ethnogynaecological study conducted in the tribal areas of North Waziristan which highlights the importance of traditional herbal remedies for their basic medical requirements. The results of this study would serve as a baseline for advanced phytochemical and pharmacological screening, as well as conservationists for further studies.

A Cross-Cultural Analysis of Plant Resources among Five Ethnic Groups in the Western Himalayan Region of Jammu and Kashmir.

Plant resources have always been valuable in human life, and many plant species are used in medicine, food, and ritual, and resource utilization is closely related to cultural diversity. Our study was conducted from June 2019 to April 2021, during which we aimed to document the local knowledge of plant resources of five ethnic groups, i.e., the Gujjar, Bakarwal, Kashmiri, Pahari, and Dogra communities of the Jammu and Kashmir (J&K) region, Western Himalayas. Through semi-structured interviews (N = 342) and group discussions (N = 38), we collected data on the ethnobotanical uses of plant resources. The data was subjected to hierarchical cluster analysis and ordination techniques (Principal Component Analysis) via, R software of version 4.0.0. Traditional uses were classified into three groups, i.e., single-, double-, and multi-use groups. The study recorded a total of 127 plant species, belonging to 113 genera and distributed among 64 botanical families. The dominant plant families were the Asteraceae, with 8% of all species, followed by Lamiaceae (6%), Polygonaceae (5%) and Ranunculaceae (4%). The recorded plant taxa were frequently used for medicine (51.4% responses), followed by food (14.9%), and fodder (9.5%). Principal component analysis (PCA) separated three groups of provisioning services depending on plant consumption preference levels. Comparative analysis showed remarkable similarities in plant uses (food, medicinal) among the Gujjar and Bakarwal ethnic groups, as both groups share a common culture. Some plants like Azadirachta indica, Brassica campestris, Ulmus wallichiana, Amaranthus blitum, and Celtis australis were also used for magico-religious purposes. We also recorded some medicinal uses that are new to the ethnobotanical literature of the J&K Himalayas, such as for Betula utilis, Sambucus wightiana, and Dolomiaea macrocephala, in our case for example local medicinal recipe, which is derived from Dolomiaea macrocephala, often known as Nashasta, used to treat weakness, back pain, and joint pain. Similarly, we also recorded new food uses for Eremurus himalaicus. Moreover, we also observed some plants for instance, Fragaria nubicola, Betula utilis and Juniperus communis have spiritual significance (i.e., amulets and scrolls) for this part of the Himalayan region. The present study provides a useful tool for resource management and can help in developing scientifically informed strategies for the conservation of plant resources.

Ethnoveterinary Practices of Medicinal Plants Among Tribes of Tribal District of North Waziristan, Khyber Pakhtunkhwa, Pakistan.

Domestic animals play a vital role in the development of human civilization. Plants are utilized as remedies for a variety of domestic animals, in addition to humans. The tribes of North Waziristan are extremely familiar with the therapeutic potential of medicinal plants as ethnoveterinary medicines. The present study was carried out during 2018-2019 to record ethnoveterinary knowledge of the local plants that are being used by the tribal communities of North Waziristan, Khyber Pakhtunkhwa, Pakistan. In all, 56 medicinal plant species belonging to 42 families were identified, which were reported to treat 45 different animal diseases. These included 32 herbs, 12 shrubs, and 12 trees. Among the plant families, Asteraceae contributed the most species (5 spp.), followed by Amaranthaceae (4 spp.), Solanaceae (4 species), and Alliaceae, Araceae, and Lamiaceae (2 spp. each). The most common ethnoveterinary applications were documented for the treatment of blood in urine, bone injury, colic, indigestion, postpartum retention, skin diseases, constipation, increased milk production, mastitis, foot, and mouth diseases.

Antidiarrheal and antispasmodic activities of Trillium govanianum rhizomes extract: involvement of calcium channel blockade.

The antidiarrheal effect of methanolic extract of Trillium govanianum Wall. ex D. Don (Melanthiaceae alt. Trilliaceae) was studied at doses of 12.5, 25, and 50 mg/kg in different animal models of diarrhea including castor oil (6 mL/kg), magnesium sulfate (2 gm/kg), sodium picosulfate (2 mL/kg) and lactitol (0.25 mL/kg). The antispasmodic effect of T. govanianum was studied on isolated rabbit's jejunum, using acetylcholine as tissue stabiliser and verapamil as calcium channel blocker. T. govanianum attenuated the diarrhea by producing a significant decrease in the number and weight of stool, and an increase in stool latency time. T. govanianum completely inhibited both spontaneous as well as high potassium induced contractions of isolated rabbit's jejunum, which was analogous to verapamil. Moreover, T. govanianum produced a right shift in calcium concentration response curve, confirming its calcium channel blocking activity. These findings provide scientific ground to its medicinal use in diarrhea and gut spasms.

In-vitro propagation and phytochemical profiling of a highly medicinal and endemic plant species of the Himalayan region (Saussurea costus).

Efficient protocols for callus induction and micro propagation of Saussurea costus (Falc.) Lipsch were developed and phytochemical diversity of wild and in-vitro propagated material was investigated. Brown and red compact callus was formed with frequency of 80-95%, 78-90%, 70-95% and 65-80% from seeds, leaf, petiole and root explants, respectively. MS media supplemented with BAP (2.0 mgL-1), NAA (1.0 mgL-1) and GA3 (0.25 mgL-1) best suited for multiple shoot buds initiation (82%), while maximum shoot length was formed on media with BAP (1.5 mgL-1), NAA (0.25 mgL-1) and Kinetin (0.5 mgL-1). Full strength media with IAA (0.5 mgL-1) along with IBA (0.5 mgL-1) resulted in early roots initiation. Similarly, maximum rooting (87.57%) and lateral roots formation (up to 6.76) was recorded on full strength media supplemented with BAP (0.5 mgL-1), IAA (0.5 mgL-1) and IBA (0.5 mgL-1). Survival rate of acclimatized plantlets in autoclaved garden soil, farmyard soil, and sand (2:1:1) was 87%. Phytochemical analysis revealed variations in biochemical contents i.e. maximum sugar (808.32 µM/ml), proline (48.14 mg/g), ascorbic acid (373.801 mM/g) and phenolic compounds (642.72 mgL-1) were recorded from callus cultured on different stress media. Nonetheless, highest flavenoids (59.892 mg/g) and anthocyanin contents (32.39 mg/kg) were observed in in-vitro propagated plants. GC-MS analysis of the callus ethyl acetate extracts revealed 24 different phytochemicals. The variability in secondary metabolites of both wild and propagated plants/callus is reported for the first time for this species. This study may provide a baseline for the conservation and sustainable utilization of S. costus with implications for isolation of unique and pharmacologically active compounds from callus or regenerated plantlets.

Evaluation of hepatoprotective effects of crude methanolic extract of Datura metel L. in mice.

Datura metel has been recommended in several human disorders including a remedy for liver toxicity. The current study was designed to evaluate the hepatoprotective effect of methanolic extract of D. metel in animal model. Acute toxicity of methanolic crude extract of Datura metel (MEDM) was studied in animals in various doses 500-2000 mg/kg. Mice of either sex were divided into groups (n=6). One group received normal saline intraperitonially as negative control, while other gentamicin 100mg/kg for 8 days as positive control. 3rd group received 50mg/kg silymarin as standard, 4th group received 100mg/kg of MEDM, 5th group received 200mg/kg MEDM while 6th group received 300mg/kg MEDM and gentamicin 100mg/kg for 8 days. The blood samples were collected on 9th day and the animals were then dissected and the liver of all the animals were isolated. MEDM was found safe in acute toxicity test at various doses up to 2000 mg/kg. The levels of serum glutamic pyruvic transaminase and alkaline phosphatase were elevated significantly with gentamicin treatment which significantly down-regulated by MEDM (100, 200 and 300 mg/kg) in a dose dependent manner.. The histological examination showed that the MEDM has markedly treated the inflammatory infiltrate, fatty changes and congested blood vessels which were induced by gentamicin.  The findings of our study thus proved the absolute of MEDM in acute toxicity test; followed by significant hepatoprotective effect in gentamicin induced hepatotoxic mice.

GC/MS analysis, anti-leishmanial and relaxant activity of essential oil of Chenopodium ambrosioides (L.) from Malakand region.

Chenopodium ambrosioides is abundantly available in Malakand region. As constituents and concentrations of essential oils vary based on its geographical location, we carried our current study to extract and evaluate its possible relaxant activity in rabbits' jejunum and anti-leishmanial activity against promastigotes of Leishmania tropica. The essential oil was obtained from aerial fresh parts through steam distillation followed by GC/MS analysis. Antispasmodic activity was performed on spontaneous and KCl induced contractions. Curves for calcium concentration response (CCRCs) were prepared with and without different concentrations of essential oils and verapamil - a standard calcium channel blocker as per our reported procedures. GC/MS analysis indicated that the essential oil contains 4-carene (56.59%) and o-cymene (41.46%), the two most abundant compounds previously reported from this species. The LD50 value for acute toxicity is 279.66±2.2mg/kg. The essential oil have significant antileishmanial activity with LC50 of Log10 (1.83±0.0026) ×10-6mg/ml, potent relaxant activity on rabbits' jejunal preparations with respective EC50 = 1.46±0.15mg/ml for spontaneous activity. For KCl (80mM) induced contractions, EC50=0.26±0.02mg/ml. In CCRCs, the oil produced a right shift as exhibited by verapamil. More, its relaxant activity, which is mediated through calcium channel blocking mechanism, proves a rationale for its traditional use in gut spasm.

Comparison of acute and chronic effects of Bacopa monnieri, Ginkgo biloba, and Lavandula angustifolia and their mixture on learning and memory in mice.

Forty-seven million people are living with memory-related disorders worldwide. Phytomedicines are gaining extensive interest in the treatment of these ailments. Memory-enhancing (acute and chronic) potentials of commercial grade extracts of Bacopa monnieri (200 mg/kg, po), Ginkgo biloba (150 mg/kg, po), and Lavandula angustifolia (200 mg/kg, po) and their mixture (B. monnieri 100 mg/kg, G. biloba 75 mg/kg, and L. angustifolia 100 mg/kg, po) were compared for their synergistic/additive effects on the Morris water maze (MWM) test and elevated plus maze (EPM) test in scopolamine-induced amnesia in mice. Escape latency and accumulative path length were significantly reduced both in acute (up to day 6) and chronic trials (days 8-14) in B. monnieri-, G. biloba-, and L. angustifolia-treated animals and their mixtures (n = 8, p < .05) in MWM. Furthermore, in probe trials (acute on day 7 and chronic on day 15), the number of crossing-overs at platform position and time spent in platform quadrant were significantly increased, while transfer latency in EPM was decreased in treated animals as compared to the saline group (n = 8, p < .05). The mixture showed synergistic effects on memory enhancement as compared to each extract individually in mice. Further studies may be carried out on the active compounds of B. monnieri at the cellular and molecular levels.

Evaluation of Cholinesterase Inhibitory Potential of Different Genotypes of Ziziphus nummularia, Their HPLC-UV, and Molecular Docking Analysis.

Ziziphus nummularia is an important source of valuable phytoconstituents, which are widely used in traditional medicine system of Indo-Pak sub-continent. In this study we investigated the distribution of phenolic compounds in the fruit pericarps of six different genotypes (ZNP01-06) of Z. nummularia growing in the unexplored hilly areas of Pakistan. The methanolic extracts of these genotypes were screened for total phenolic content (TPC), total flavonoid content (TFC), antioxidant, and cholinesterase inhibitory potentials. The observed biological potentials were explained in terms of the outcome of molecular docking and HPLC analyses. Among them, genotype ZNP02 displayed high TPC (88.50 ± 1.23 µg/mL) and showed potent scavenging activity against DPPH (67.03 ± 1.04 µg/mL) and ABTS (65.3 ± 1.74 µg/mL) in comparison to ascorbic acid (68.7 ± 0.47 µg/mL). Moreover, genotypes ZNP01, ZNP02, and ZNP04 displayed potent inhibition against acetyl and butyryl cholinesterases (AChE and BChE) with IC50 values of 21.2, 20.5, and 23.7 µg/mL (AChE) and 22.7, 24.4, and 33.1 µg/mL (BChE), respectively. Furthermore, the individual compounds in the most potent species ZNP01 responsible for potent enzyme inhibition (identified through HPLC-UV analysis), were computed via docking simulation software to the enzyme structures. Among these compounds rutin exhibited significant binding affinity with value of -9.20 kcal/mol. The differences amongst the phytochemical compositions of the selected genotypes highlighted the genotypic variations in them. Based on our results it was concluded that the selected plant can be used as remedy of oxidative stress and neurodegenerative diseases. However, further studies are needed to isolate responsible compounds and test the observed potential in vivo, along with toxicological evaluations in animal models.

Relaxant activity of different fractions of fruits of Rosa moschata J.

The current work is an attempt to know that in which fraction(s) the relaxant constituents of Rosa moschata concentrate. Crude methanolic extract of Rosa moschata was prepared as per our reported procedure. Sub fractions of methanol extract were extracted with different solvents in increasing order of polarity i.e. n-hexane > chloroform > ethyl acetate > n-butanol > residual aqueous fractions. Different concentrations (0.01, 0.03, 0.1, 0.3, 1, 3, 5 and 10 mg/ml) of the fractions were tested on spontaneous contractions and KCl induced contractions on rabbits' jejunal preparations. Calcium Concentration Response Curves (CCRCs) in the presence and absence of the test fractions using verapamil were constructed to understand its mechanisms. EtOA fraction was more relaxant with EC50 values 0.812±0.149 mg/ml on spontaneous and 2.01±0.08 mg/ml on KCl induced contractions. we also found right shift in its EC50 values expressed as log [Ca++]M values. In presence of 0.3 mg/ml EtOA fraction, its EC50 value was -2.22±0.035 vs control EC50 -2.71±0.21. For n-BuOH fraction, EC50 value was -1.82±0.00 vs control with EC50 -2.28±0.049 at concentration of 0.3 mg/ml. Ethyl acetate fraction of Rosa moschata was more potent and is therefore can be a target for activity guided isolation of calcium channel antagonists.

Nephroprotective effects of Datura metel extract in gentamicin induced mice model: biochemical and histological evidences.

Datura metel is traditionally used as a remedy for renal toxicity. However, the nephroprotection has not been scientifically validated yet. To evaluate the nephroprotective like effect of methanolic extract of D. metel in gentamicin induced mice model, mice of either sex were divided into groups. One group received normal saline as negative control. The 2nd group received gentamicin 100mg/kg for 8 days as positive control, 3rd group received 50mg/kg silymarin as standard, while the reaming groups received 100, 200 and 300 mg/kg of MEDM and gentamicin 100mg/kg, for 8 days. The blood and urine samples were collected on 9th day, animals were then dissected and whole kidneys were removed and preserved in formalin for later histological examinations. The level of serum creatinine, blood urea nitrogen, urine creatinine and urine urea were significantly (P<0.05) elevated and the renal MDA level was also elevated significantly (P<0.05) by gentamicin in mice. After the treatment of test animals with MEDM, the elevated level of serum and urine biomarkers by gentamicin were reversed by MEDM. The nephroprotective effect was found in dose dependent manner. As the MEDM significantly protected the nephrotoxicity via its antioxidant effect. The findings of our study thus proved the scientific background for the nephroprotective effect of MEDM.

Herbal Teas and Drinks: Folk Medicine of the Manoor Valley, Lesser Himalaya, Pakistan.

In spite of the remarkable achievements in the healthcare sector over recent decades, inequities in accessibility and affordability of these facilities coexist throughout Pakistan. Thus, we aimed to explore and document the cultural knowledge of herbal teas used medicinally by the local community members of Manoor Valley, Pakistan. Field investigations were undertaken during the summer season of 2015-2017, and cultural practices of medicinal plant usage for treating various ailments were gathered through interviews of the local inhabitants. Ethnomedicinal insights of the medicinal plants used in herbal teas were gained with different indexes. Our results revealed 27 plant species, comprising of herbs (70%), shrubs (26%), and trees (4%), which were used for treating 21 diseases. Plants belonged to 18 families: Asteraceae and Lamiaceae were the leading families used for treating diseases. Diarrhea and gas troubles were the most frequent diseases. Based on indexes values, Cannabis sativa was the dominant species used. The results revealed that 57% of medicinal uses are new to literature. This ethnomedicinal study is providing the first insights into the traditional medication system of Lesser Himalaya, Pakistan, through ethnomedicinal teas.

Exploring threatened traditional knowledge; ethnomedicinal studies of rare endemic flora from Lesser Himalayan region of Pakistan

ABSTRACT Himalayas are one among the world biodiversity hotspots harboring many endemic medicinal plants. Despite augmentation in the documentation of ethnopharmacological knowledge of medicinal plant species, information regarding endemic species is still underway. Current paper highlights the traditional medicinal uses of rare endemic and unexplored group of plants having potential for novel chemical constituents with effective pharmacological activities. In total, 142 informants (91 male and 51 female) including seventeen traditional healers were interviewed using semi-structured questionnaire, personal observations and group discussions. Interviews were taken in field or otherwise photographs were shown for identification. Females were interviewed indirectly through male family members. For data analysis, quantitative analytical approach was adopted using ethnopharmacological indices as Relative frequency of citations and Fidelity Level. In total, 38 endemic plant species belonging to nineteen families were utilized by the local inhabitants. Highest number of endemics was belonging to Ranunculaceae (7), followed by Gentianaceae and Rosaceae (4 each) with respect to number of species. Highest number of endemics was used in fever, wound healing, throat infection and tonic (4 species each). Root was the most widely used part (36.17%) in cure of diseases and the leading mode administered was decoction (25.49%). Highest use reports and RFC values were recorded for Pimpinella stewartii (58 citations, 0.41 RFC), Caltha alba var. alba (52 citations, 0.37 RFC). Endemic plant species considerably contribute toward ethnomedicinal knowledge and despite rarity, the communities prefer their utilization. Conservation of endemics is necessary for future availability to the local communities.

A possible mechanistic approach of synthetic flavonoids in the management of pain.

Flavonoids are phenolic compounds that have always attracted pharmaceutical researchers and food manufacturers. Nature has indirectly provided us flavones in our daily diet i.e. tea, fruits, juices and vegetables. Flavones have got special position in research field of natural and synthetic organic chemistry due to their biological capabilities. Three substituted flavone derivatives have been synthesized from substituted O-hydroxy acetophenones and 4-trifluoromethyl benzaldehyde in good yield. The structures have been established by different spectroscopic techniques like 1HNMR 13CNMR, IR spectroscopy. The compounds were then screened for their enzyme inhibition potential and antinociceptive response in mice models with writhings induced by acetic acid, tail immersion and formalin-induced nociception assay procedures and structure activity relationship was established. The effects following pretreatment with naloxone were also studied to reveal the involvement of opioid receptors in the antinociceptive action. The flavone derivatives showed moderate to weak inhibition against LOX. Moreover, significant to moderate decrease in the number of abdominal constrictions, increase in paw-licking response time in both phases and a significant raise in latency time in nociception models. Moreover, the antinociceptive response was significantly attenuated by pretreatment with opioid receptor antagonist suggesting the involvement of opioidergic system in the analgesic action. The flavone derivatives showed analgesic response in all models of nociception suggesting the possible involvement of opioidergic system in the antinociceptive action.

Historical perspectives of ethnobotany.

Ethnobotany is the study of the direct interaction between plants and man in his culture. From time immemorial, man has been depending on Mother Nature for all his basic needs and plant diversity that existed around him always attracted his curiosity. Man's preliminary interest in plants started from his need for food, shelter, protection and then his attention shifted to the remedies for injuries and diseases. In fact, medicine and botany always had strong and close connections, as most of the modern medicines come from plant sources. The current venture reviews the developments and perspectives of ethnobotany in global scenarios. We have summarized the available literature and documentations regarding the historic background of ethnobotany. A survey of the literature survey reveals that plant resources were cited around the globe for curing almost all disease categories i.e. skin disorders, respiratory disorders, digestive disorders, urinary disorders, cardiac disorders, ophthalmic disorders, ear nose throat (ENT) disorders, excretory disorders, nervous disorders, immunity disorders and so on. The orally exchanged customary information/knowledge is in the hands of elders, and most of it can vanish after their deaths and could lead to the depletion of such imperative information. The ethnic society resources, like the folk asset in association with a nature of investigated and surveyed areas, can be conserved through documentation. The natural chemical compounds from plants may be screened and isolated. Plants with therapeutic uses should be tested in studies by using those isolated natural compounds in laboratories to screen and evaluate of plants metabolites with potential therapeutic uses.

Ethnoveterinary Therapeutic Practices and Conservation Status of the Medicinal Flora of Chamla Valley, Khyber Pakhtunkhwa, Pakistan.

Domestic animals play a very important role in the human civilization. Besides human being, plants are used as medicines for many domestic animals. The therapeutic practices are very common among the tribes of Chamla, rich in ethnoveterinary medicinal plants. Due to poor availability of modern healthcare facilities and poverty of indigenous people, they depend on local medicinal plants for the healthcare of their domestic animals. This study is the first attempt to document the indigenous knowledge and evaluate the conservation status of medicinal plants and practices of herbal remedies by the local people of Chamla Valley in the treatment of their livestock. Semi-structured questionnaire was used and 120 local inhabitants were interviewed to note the traditional practices regarding plant species uses. Well-known statistical indices, Use Value formula and Relative Frequency Citations were used for quantification of the recorded data. It was observed that 50 medicinal plants belonging to 38 families were reported, where Poaceae was the most cited. The common livestock are goats, sheep, buffalos, cows, bulls, and donkeys. Most of the herbs, which are used in livestock treatment, are wild and few plants are cultivated. The common livestock diseases are red water, 3 days sickness, diarrhea, tympany, and indigestion among others. Most of the plants are used in fresh condition. According to the results, Brassica nigra was used for placenta retention, Butea monosperma for constipation, Calotropis procera for indigestion and 3 days sickness. Canabis sativa, Cedrella serrata, Allium sativum, and Origanum vulgare were used for fever. The traditional plant collection techniques have resulted in huge losses of these valuable plant resources. The ethnobotanical conservation assessment revealed that due to increased exploitation and un-sustainable harvesting, 49% of these economically valued medicinal plant species are decreasing in last 30 years. Some of the plants are only present on high altitudes while they had been finished in the foothills like Paeonia emodi and Berberis lycium. Lack of scientific knowledge, ignorance, poverty, and joblessness, as well as land development, construction and fires, add more pressure on flora and fauna of the area and various species are under the threat of extinction.

Antidiabetic potential of flavonoids from Artemisia macrocephalla Jaquem in streptozotocin-induced diabetic rats: Pharmacological and biochemical approach.

Plants belongs to Asteraceae family are reported to be rich in major phytochemical including flavonoids and are documented to acquire antidiabetic response. Antidiabetic effects of salvigenin, eupatilin and cirsilineol were screened on in-vitro enzyme inhibition and in-vivo streptozotocin animal models. Administration of salvigenin, eupatilin and cirsilineol (7.5 and 15mg/kg) produced antidiabteic responses in streptozotocin model for diabetes. All natural flavonoids reduces the blood glucose level to a significant level (*P<0.05, **P<0.01, ***P<0.001, n=8) but promising results were observed in eupatilin at dose of 7.5mk/kg (364.12±4.3 to 128.41±4.2mg/dL, n=8) and at dose of 7.5mk/kg 363.65±4.8 to 126.14±5.1mg/dL, n=8). Administration of salvigenin, eupatilin and cirsilineol (7.5 and 15mg/kg) for 28 days showed a substantial fall (*P<0.05, **P<0.01, ***P<0.001, n=8) in total cholesterol, LDL and triglcerides (TGs) in comparison to diabetic model. The isolated flavonoids reduced considerably the serum ALP, SGPT and SGOT in rats intoxicated with streptozotocin. The results indicate that the flavonoids may be useful in the development of new antidiabetic drugs.

Evaluation of safety profile and stress suppressant activity of Rosa moschata in mice.

Stress is a state that seriously disturbs psychological or physiological homeostasis of the body and subsequently affects the morphology and function of the hippocampus. Currently available anti-stress medications provide limited benefits with cost of severe adverse effects. In the present study, effect of Rosa moschata extract was evaluated using acute restraint model in mice. The stress suppressant activity of Rosa moschata was evaluated by using elevated plus maze test (EPM), dark light box test and open field test (OFT) following restraint stress protocol. Results showed that the Rosa moschata extract significantly enhanced the number of transitions and the time spent in the open arm in the EPM, increased the number of transitions and time spent in the light compartment of the dark light box, and also enhanced the locomotor activity in OFT, as compared to the stress group. In addition, LD50 of the plant extract is greater than 5000mg/Kg. Thus the findings of our studies show that Rosa moschata significantly alleviates stress following the acute restraint stress in mice. Further studies dealing with underlying mechanism and characterization of active fraction/compound may provide an alternative therapy for stress and related neurological conditions.

Isolation of flavonoides from Artemisia macrocephala anticholinesterase activity: Isolation, characterization and its in vitro anticholinesterse activity supported by molecular docking.

In this study the flavonoids isolated from Artemisia macrocephala were screened out for anticholinesterase activity. The isolated flvanoids were characterized by HNMR, NOESY, COSY, HMBC, HSQC and mass spectroscopy. The compounds (1-4) in appropriate quantities were isolated from chloroform fraction using gravity column chromatography by eluting ethyl acetate/n-hexane solvent system. The flavonoids were characterized and resulted in the form of mono substituted methoxy flavones to tri substituted flavones. Ellman's assay techniques were used to find out enzyme inhibition. Operating environment (MOE) software was used for molecular docking studies. Compounds (1), (2) and (3) showed 88.42±2.76, 84.50±1.60 and 90.16±2.98 percent inhibition of the acetyl cholinesterase (AChE) respectively at 1000µg/mL concentrations with IC50 value 165, 60, 65µg/mL respectively which were comparable to that of standard galanthamine. While for butyryl cholinesterase (BChE), (1), (2) and (3) showed 91.63±4.32, 81.03±3.53 and 87.69±2.84 percent inhibitions respectively at 1mg/mL as compared to the standard galanthamine which caused 96.50±2.41 percent inhibition at the same concentration. Whereas, compound (4) exhibited moderate activity for both the enzymes. Molecular docking studies confirmed the experimental AChE and BChE inhibitory activities of the test samples by their virtue of multiple teractions with target enzymes. The results confirm that the specie has biologically active constituents that are more useful for the management of several neurodegenerative ailments like ataxia, Parkinson's disease, Alzeimer's disease and some other types of dementia.