Neurotoxicity, antipsychotic and antidepressant screening of the fruit of Rosa moschata in mice.

Extract of Rosa moschata (RM) fruits was evaluated for the anti-schizophrenic and antidepressant activities. We first determined the neurotoxic effect of hydro-methanolic extract of RM using inverted-screen test. Further, the extract was tested in the ketamine-induced schizophrenia model and its antidepressant effect was assessed by tail suspension and forced swim test in mice. Different doses of extract were administered once/day to the animals for 14 consecutive days. Behavioral parameters were investigated 24h after last administration of drug/extract by performing Y-maze test, forced swim test and open field test. Results showed that TD50 of the extract was ~1000mg/Kg. Moreover, extract significantly increased % alternations in YMT, reduced immobility time in FST and enhanced locomotion in OFT compared to saline group. Similarly, RM extract decreased time of immobility in FST and TST significantly showed antidepressant effect. Thus, it was concluded that extract of RM has antipsychotic and antidepressant properties.

Dirty versus clean fuel for cooking in Pakistan: regional mapping and correlates.

Based on the Pakistan Demographic and Health Survey (PDHS) 2017-18 data, the study intends to present the relative status of a particular fuel (type) in terms of its level of use in different regions of Pakistan. At the same time, the study investigates the determinants of the household type of fuel used for cooking in Pakistan. Socioeconomic characteristics of the households and household heads are taken as determinants of the main type of fuel (clean or dirty) used for cooking by the households. Binary logistic regression is applied as an estimation technique. Over the country, wood stands first as the most widely used type of fuel followed by natural gas followed by LPG. Animal dung and charcoal are respectively ranked as the fourth and fifth most commonly used types of fuel for cooking by households in Pakistan. In comparison amongst the four provinces of Pakistan, the use of wood as well as charcoal as the fuel type for cooking (in relative terms) is the most common in Balochistan province. Likewise, Punjab province is the most frequent user of liquefied petroleum gas (LPG), crop residues, and animal dung, while Sindh province is the most frequent user of natural gas. Households possessing agricultural land and livestock and with large household sizes had more probability to use dirty sources of fuel for cooking. Households with better wealth status and residing in urban areas were more likely to use clean sources of fuel for cooking. Household head's higher educational status and male-gender had more likelihood to use clean sources of fuel for cooking.

Medicinal waterbirds in the traditional healthcare system: an assessment of biodiversity-cultural linkages in Eastern Khyber Pakhtunkhwa, Pakistan.

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

Black phosphorus nanosheets/poly(allylamine hydrochloride) based electrochemical immunosensor for the selective detection of human epididymis protein 4.

In this work, an electrochemical immunosensor based on black phosphorus nanosheets (BPNS)/poly(allylamine hydrochloride) (PAH) nanocomposite modified glassy carbon electrode was developed for the detection of ovarian cancer biomarker HE4. PAH has been applied to retain BPNS in its original honeycomb structure and to anchor biomolecules electrostatically on the transducer surface. The as synthesized nanocomposite was characterized by zeta potential analysis, scanning electron microscopy, x-ray photoelectron spectroscopy, transmission electron microscopy, high-resolution transmission electron microscopy. Subsequently, the performance of the electrochemical immunosensor was evaluated through cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy. Under the optimal condition, the developed electrochemical immunosensor permitted to detect HE4 with a linear range of 0.1-300 ng ml-1and a detection limit of 0.01 ng ml-1. The developed sensor exhibited good selectivity and specificity to HE4 with negligible interference effect from common biomolecules like bovine serum albumin, lysozyme, protamine, glucose, fructose, hemoglobin and fetal bovine serum. Further, practical application of developed electrochemical immunosensor was demonstrated in spiked human serum which showed satisfactory recovery percentages.

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.

Anticonvulsant activity of methanolic extract of Withania cogulans in mice.

Mental and neurological diseases including depression, Parkinson's disease, dementia, epilepsy, anxiety disorders and bipolar disorders account for a considerable amount of the world's disease burden. Unfortunately, drugs used in the treatment of neurological diseases are expensive, symptomatic and they produce undesirable side effects. People from different cultures prefer to use medicinal plants for the treatment of various ailments ranging from plain to perplex disorders because they are most affordable, cost effective and easily accessible source of treatment in the primary healthcare system throughout the world. Withania coagulans, an erect grayish under-shrub belongs to family Solanaceae. It is common in Pakistan, East India, Iran and Afghanistan. The objective of this study was to analyze the anti-seizure activity of crude methanolic extract of Withania coagulans fruits (MeWc). For screening of this activity, maximal electroshock seizures model (MES) and chemically-induced seizures models were used. In maximal electroshock seizures test MeWc showed significant dose dependent percent protection against hind-limb tonic extension; significant and dose-dependent increase in latency to myoclonic jerks and tonic clonic convulsions and decrease in seizures duration were observed in PTZ-induced seizures. In strychnine-induced convulsions MeWc significantly increased latency to hind-limb tonic extension and percent protection from death in a dose-dependent manner. Thus, it was inferred from the experiments that extract of Withania coagulans showed anticonvulsant activity.

An ethnobotanical study of wetland flora of Head Maralla Punjab Pakistan.

BACKGROUND: Wetlands are biologically diverse and highly productive ecosystems that support one-third of all threatened and endangered plants of the world. Wetland plants have been studied ethnobotanically much less than terrestrial plants, including in Pakistan, thus information about the uses of local wetland plants in traditional healthcare system is scare. Head Maralla is a non-recognized wetland with diversified flora that has been focused of the current study. METHODS: The ethnobotanical data were collected from four sites viz., River Tavi, Upstream Chenab, River Manawarwala Tavi, and Bhalolpur through questionnaire and interviews during field trips. Quantitative indices including informant consensus factor (ICF), cultural significant indext (CSI), relative frequency of citation (RFC), and use value (UV) were used to analyze the data. RESULTS: On the whole, 119 plant species were identified belonging to 54 families, of which 87 species were dicot, 12 monocots, 11 aquatic, 5 ferns, and 4 species were bryophytes. Of these, 50% of the plant species were utilized for therapeutic purposes, followed by leaves which had more than 20% usage of total consumption. Herbs were the primary source of medicine (73 spp) followed by trees (22 spp), weeds (11 spp), shrubs (9 spp), foliose (2 spp) and thaloids (2 spp) in the area. Fic ranged from 0.66 to 0.93 for constipation and respiratory disorders with an average Fic of 0.87 reflecting a high consensus among the informants about the use of plants to treat particular ailment. Major ailments viz., urination (14%), cough (8.40%), cold (6.70%), stomach (5.90%), asthma (5.90%), skin infection (5%), constipation (5%), and diarrhea (4%) etc., were treated with local plant recipes. The highest CSI value was found for A. vesica (7.55) widely used in respiratory disorders and in digestive problems. RFC ranged from 0.92 to 0.15 with the maximum value obtained for R. communis (0.95). The use values ranged from 0.03 to 0.90 with the maximum use value for R. communis (0.90). A positive correlation was found between CSI and RFC (r = 0.29), and CSI and UV (r = 0.29). The JI values ranged from 7.14 to 0.14 indicating strong affinity with Samahani valley, Azad Kashmir, Pakistan. Unique species Osmunda regalis was first time reported from Pakistan with novel uses for renal and blood purifier. Fifteen percent (15%) plants contribute as fodder species consumed by local community for livestock while almost 6.7% species were utilized for timber and fuel purposes. CONCLUSION: The ecosystem of Head Maralla provide a complex habitat for aqauatic, terrestrial, and agriculture wetland vegetation. It is suggested that conservation efforts should be made to conserve the ethnoecological knowledge of these areas and pharmacological studies should be conducted for noval drug synthesis in future.

Exploring the genetic diversity and population structure of scarlet eggplant germplasm from Rwanda through iPBS-retrotransposon markers.

BACKGROUND: Scarlet eggplant (Solanum aethiopicum gr. gilo) is a part of African indigenous vegetables and acknowledged as a source of variations in the breeding of Brinjal. Since its genetic diversity is still largely unexplored, therefore genetic diversity and population structure of this plant were investigated in this study. METHODS AND RESULTS: Scarlet eggplant germplasm made of fifty-two accessions originated from two districts of Rwanda was assessed by employing the iPBS-retrotransposon markers system. Twelve most polymorphic primers were employed for molecular characterization and they yielded 329 total bands whereupon 85.03% were polymorphic. The recorded mean polymorphism information content was 0.363 and other diversity indices such as; mean the effective number of alleles, mean Shannon's information index and gene diversity with the following values; 1.298, 0.300 and 0.187 respectively. A superior level of diversity was noticed among accessions from Musanze district. The model-based structure, neighbor-joining, and principal coordinate analysis (PCoA) gathered scarlet germplasm in a divergence manner to their collection district. Analysis of molecular variance (AMOVA) displayed that the utmost variations (81%) in scarlet eggplant germplasm are resulting in differences within populations. CONCLUSIONS: The extensive diversity of scarlet eggplant in Rwanda might be used to form the base and genetic resource of an exhaustive breeding program of this economically important African indigenous vegetable. For instance, accessions MZE53 and GKE11 might be proposed as parent candidates due to their high relative genetic distance (0.6781).

Systems pharmacology-based dissection of mechanisms of Tibetan medicinal compound Ruteng as an effective treatment for collagen-induced arthritis rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Compound Ruteng (CRT) is a prescribed formulation based on the theory of Tibetan medicine for the treatment of yellow-water-disease. It is consisted with 7 medicinal material include Boswellia carterii Birdw (named "Ruxiang" in Chinese); Tinospora sinensis (Lour.) Merr. (named "Kuan-Jin-Teng" in Chinese), Cassia obtusifolia L (named "Jue-Ming-Zi" in Chinese); Abelmoschus manihot (L.) Medic (named "Huang-Kui-Zi" in Chinese); Terminalia chebula Retz. (named "He-Zi" in Chinese); Lamiophlomis rotata (Benth.) Kudo (named "Du-Yi-Wei" in Chinese) and Pyrethrum tatsienense (Bur. et Franch.) Ling (named "Da-Jian-Ju" in Chinese). They are widely distributed in Tibet area of China and have been used to treat rheumatism, jaundice, and skin diseases for centuries. AIM OF THE STUDY: The present study was conducted to investigate the anti-arthritis effect of CRT and to disclose the systems pharmacology-based dissection of mechanisms. MATERIALS AND METHODS: The chemical constituents in CRT were identified using HPLC method, and CRT candidate targets against RA were screened by network pharmacology-based analysis and further experimentally validated based on collagen-induced arthritis (CIA) rat model. Furthermore, therapeutic mechanisms and pathways of CRT were investigated. RESULTS: 391 potential targets (protein) were predicted against 92 active ingredients of 7 medicinal materials in CRT. Enrichment analysis and molecular docking studies also enforced the practiced results. X-ray based physiological imaging showed the attenuated effect of CRT on paw swelling, synovial joints and cartilage with improved inflammation in CIA rats. Moreover, the expression of biomarkers associated with RA such as MMP1, MMP3 and MMP13 and TNF-a, COX2 and iNOS are down-regulated in ankle joints, serum, or liver. CONCLUSION: In conclusion, CRT compound could attenuate RA symptoms and active ingredients of this compound could be considered for drug designing to treat RA.

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.

Development and Standardization of Moringa oleifera Leaves as a Natural Dietary Supplement.

Moringa oleifera leaves were selected as a model due to their hundreds of health benefits. On the other hand, the powder of these leaves has exhibited poor flowability, low tensile strength, bitter taste, poor dissolution rate, and lack of information regarding dosage. These are the common hurdles and limitations in the adaptation of herbal-based medications. Therefore, a comprehensive study was planned to introduce herbal-based medicines into mainstream medicines by standardization according to the U.S. Food and Drug Administration (FDA) and international pharmaceutical standards. A Simplex Lattice Design (SLD) of Design Expert 8.0 software was used to formulate different concentrations of superdisintegrant, binder/diluent, and sweeteners. An Instron Universal Testing machine coupled with a 13 mm stainless cylindrical die was used to manufacture tablets by means of direct compression method at 20 kN applied force. Therefore, selection of excipients was made on the basis of their tensile strength, flowability, and taste-masking properties. Optimum formulation was tested on rabbits for toxicity and growth rate. All formulated tablets were evaluated on standard parameters for orally disintegrating tablets described by the Food and Drug Authority (U.S.). The optimum formulation fulfills all standard parameters such as hardness, disintegration time, friability, and dissolution rate. The present formulation showed no toxicity when tested on rabbits. The present study provides a fundamental understanding of the tableting characteristics of natural medicines. The present study provides information that will help to overcome the challenges.

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.

Green Synthesis of Silver Nanoparticles Using Jurinea dolomiaea and Biological Activities.

Green syntheses of nanoparticles using plant materials are of tremendous scope. Here we report advantageous green synthesis for silver nanoparticles (AgNPs) using aqueous-root extract of Jurinea dolomiaea and AgNO3. Color change of solution and UV-Vis absorption at 444 nm indicated the formation of AgNPs. XRD confirmed their face centered cubic structure (fcc) with average particle size of 24.58 nm. SEM analysis showed their spherical, cubic and triangular structures. FT-IR indicated the presence of functional groups of reducing and stabilizing phytochemicals. Methanol-root extract of J. dolomiaea revealed high flavonoid (445 mg RE/g) and phenolic contents (92 mg GAE/g). Methanol-extract showed high antioxidant potency (IC50 = 0.494 µg/mL), rationally due to its high phenolic and flavonoid contents. These AgNPs showed the highest and equal antimicrobial activities against Escherichia coli and Pseudomonas aeruginosa (Inhibition zone 11.0 mm) whereas, methanol-roots extract showed equal and intermediate activities (Inhibition zone 8.0 mm) against both pathogens but aqueous extract showed poor activities (Inhibition zone 6.0 mm) against these both pathogens. AgNPs are playing a major role in the field of nanotechnology and nanomedicine due to their antimicrobial and drug delivery efficacy as well as reasonable tolerance in human biology.

Biogenic Synthesis of AgNPs with Saussurea lappa C.B. Clarke and Studies on Their Biochemical Properties.

Nanotechnology has become an irreplaceable need and green synthesis of nanoparticles offers several advantages over physical and chemical methods. Medicinal plants are the main reservoirs of drugs and drug candidates. We report the biogenic synthesis of silver nanoparticles (AgNPs) using aqueous root extract of Saussurea lappa. Verification and characterization of these nanoparticles were done by UV-visible spectroscopy, XRD-analysis and Scanning Electron Microscopy and FT-IR. Extract-loaded-AgNPs showed the highest inhibition zone against Escherichia coli (11.0 mm) and intermediate against Pseudomonas aeruginosa (9.0 mm). The methanolic root extract of S. lappa alone, also moderately inhibited Pseudomonas aeruginosa (9.0 mm) and showed lower activity (6.0 mm) against Escherichia coli. Its aqueous roots extract inhibited (6.0 mm) the growth of tested organisms. Methanolic extract showed antioxidant potency (IC50 = 0.814 µg/mL). Experiments revealed the presence of phenols and flavonoids in the roots of Saussurea lappa. These findings provide promising interest to exploit Saussurea lappa for the biogenic synthesis of AgNPs and their biological applications.

Hepatoprotective activity of iridoids, seco-iridoids and analog glycosides from Gentianaceae on HepG2 cells via CYP3A4 induction and mitochondrial pathway.

Gentianaceae herb extracts have been widely used as food additives, teas or medicinal remedies for various diseases and disorders of the human body. Herein, the potential effects of iridoids, seco-iridoids and analog glycosides from gentian on acontine-induced hepatotoxicity were investigated in HepG2 cells to obtain metabolic data of drug-biotarget interactions. Molecular docking analysis was performed to assess the binding efficiencies of 53 iridoids, seco-iridoids and analog compounds obtained from 50 gentian species to the active sites of human CYP3A4 enzyme. The docking scores of 29 iridoids, seco-iridoids and 24 analog glycosides were calculated from the free energy of ligand-protein complexes using a computer-assisted docking simulation. After comprehensive evaluation, 6 of these compounds, i.e., gentiopicroside, sweroside, swertiamarin, loganic acid, 6-O-ß-d-glucosyl-gentiopicroside and amarogentin were selected to evaluate their hepatoprotective effects. Quantitative real-time PCR was used to measure the expression levels of CYP3A4 mRNA in HepG2 cells. Amarogentin displayed the most clear inductive effect on CYP3A4 mRNA levels in the HepG2 cells. Moreover, amarogentin was further studied for acontine-induced toxicity in the HepG2 cells to determine the potential mechanisms. Amarogentin displayed obvious inductive effect on CYP3A4 mRNA levels in the HepG2 cells. These results elucidated that the hepatoprotective effects were caused by the facilitation of drug metabolism, amelioration of mitochondrial dysfunction and reduction of oxidative stress. Our data demonstrated that the naturally found iridoids, seco-iridoids and analog glycosides in gentian may be responsible for the hepatoprotective effects of gentian-extracted compounds and thus, this study may be useful in the food industry or in clinical practice.

Graphical dataset on important medicinal plants used for curing dental issues in Manoor Valley, Mansehra, Pakistan.

The graphical dataset in this paper is related to the research article entitled "A novel survey of the ethno medicinal knowledge of dental problems in Manoor Valley (Northern Himalaya), Pakistan" (I.U. Rahman, F. Ijaz, Z. Iqbal, A. Afzal, N. Ali, M. Afzal, M.A. Khan, S. Muhammad, G. Qadir, M. Asif, 2016) [1]. This article describes how the local community of Manoor Valley practices cultural / traditional knowledge for dental problems. For the recorded data of 25 medicinal plants, six quantitative ethnomedicinal statistical approaches / equations were used. Out of these indices, four were used to measure the most imported and cited medicinal plant species while two for the comparative analysis to evaluate the novelty of work.

A novel survey of the ethno medicinal knowledge of dental problems in Manoor Valley (Northern Himalaya), Pakistan.

ETHNOPHARMACOLOGICAL RELEVANCE: In the study area, the traditional knowledge and cultural practices of utilizing medicinal plants in the human healthcare is primarily restricted to the old age members of the community, while younger generation is mostly unaware of the natural wealth usage. AIM OF THE STUDY: We provide the first ever ethno medicinal insights into the management of dental disorders in Pakistan. The prime objective of the study was to explore novel knowledge of these local and remote community members and share it with rest of the world in documented form. MATERIALS AND METHODS: Field surveys were arranged for collection and documentation of medicinal plants of Manoor valley during 2015. Total 71 local inhabitants were interviewed randomly through questionnaire. The data obtained were quantitatively analyzed using the use value (UVi), relative frequency citations (RFCs), fidelity level index (FL%) and consensus index (CI%). For novel uses all plants were thoroughly checked with previously published articles on the same disorder and analyzed through Jaccard index (JI) and Sorensen's similarity index (QS). The collected specimens after confirmation were submitted to the Herbarium, Department of Botany, Hazara University, Mansehra, Pakistan (HUP). RESULTS: In present study 25 plants belonging to 17 families are reported, these plants were used commonly as ethno medicine. There were herbs (64%), shrubs (20%) and trees (16%) whereas, Lamiaceae with 4 species was the leading family. Majority of medicinal plant species were used in treatment of toothache (38%), wound healing (19%), gum infections (16%) whereas and lowest percentage of species were used for mouth ulcer as well as teeth powder (3%). Leaves were the most widely part used plant part in the treatment of different diseases (36%). Dominated medicinal plants with high used values were Juglans regia (UVi=0.94), Rumex dentatus (0.89) and Indigofera heterantha (0.88). Based on the RFC values, the most valuable and cited medicinal plant species used by the traditional drivers are Juglans regia having (RFCs=0.69) and Rumex dentatus (0.58). CI results show that most respondents percentage was for Juglans regia having CI%=69%, followed by Rumex dentatus (57.7%). Medicinal plants with high fidelity level and reported for one dental disorder were Berberis lycium, Geranium wallichianum, Lathyrus aphaca, Platanus orientalis, Sedum spp. and Urtica dioica for toothache. Further, new medicinal uses of Abies pindrow, Ajuga integrifolia, Berberis lycium, Urtica dioica, Indigofera heterantha were recorded for the first time from Pakistan and Astragalus grahamianus, Corydalis cornuta, Prunella vulgaris, Ranunculus muricatus, Rumex hastatus, Stellaria media, Tagetes minuta, Taraxacum officinale and Lathyrus aphaca, across the world for the current reported medicinal uses. All mentioned plant species are reported for the first time ever for dental disorders from Manoor Valley. Nonetheless, this is a novel study as no single ethno medicinal study on dental disorder up-till now has been reported from Pakistan. CONCLUSION: The present study is the first ever documentation of ethno medicinal practices aiming at the dental disorders in Pakistan, which resulted in 64% of medicinal uses new claims. So, the reported species of remote valley should be further evaluated for proper experimentation and pharmacological activities to authenticate their current traditional usage. Field observation revealed vegetation of the area was generally threatened due to its unwise use by the local communities. Trends like deforestation, over grazing, habitat fragmentation and introduction of the exotic taxa were the visible threats. So conservation of such an important natural God gifted treasure is mandatory.

Borneol inhibits TRPA1, a proinflammatory and noxious pain-sensing cation channel.

Borneol, a natural product isolated from several species of Artemisia, Blumea and Kaempferia, has a widespread use in traditional medicine. TRP ion channels are a class of nonselective cation channel proteins involved in a variety of physiological and pathological processes in mammals. TRPA1, a member of TRP family of cation channels, is involved in plethora of processes including noxious-cold, noxious-pain sensations, inflammation and the detection of irritant chemicals. Borneol is chemically related to camphor (a known inhibitor of TRPA1 ion channels); therefore, it is beneficial to investigate the effects of borneol on TRPA1. In the present investigation it was found that borneol inhibits TRPA1 mediated cationic currents in low millimolar range (IC50 0.3mM) in heterologous expression systems like Xenopus oocytes and in neurons cultured from trigeminal ganglia. Effects of nicotine, a known chemical irritant and agonist of TRPA1 are also inhibited by borneol in both systems. It is concluded that borneol, being an inhibitor of TRPA1, could be a safer therapeutic-combination in clinical situations where TRPA1 channelopathies like neuropathic-pain, trigeminal neuralgia or nicotine withdrawal treatments are involved.

Camphor modulates TRPV3 cation channels activity by interacting with critical pore-region cysteine residues.

TRPV3 ion channels mediate thermo-transduction, nociception, inflammation and dermatitis in mammals. TRPV1-4 proteins have been shown to have conserved cysteine-residues in the pore-forming regions. These residues participate in channel activation via S-nitrosylation of channel proteins. Camphor is a commonly used ligand for TRPV3 channels. Thus the knowledge about the potential binding/interacting site(s) for camphor will help to design effective and potent analgesic compounds. In an overlap-extension PCR method, following primer-pairs were used to mutate conserved cysteine-residues in the pore-region of TRPV3 channels; GATTGAGAATcCTCCAAGGACAAAAAGGAC, TRPV3-C612S-Fw and GTCCTTGGAGgACTTCTCAATCAGTCAGTGAGG, TRPV3-C612S-Rv primers pair. And for TRPV3-C619S: GGACTCcAGTTCCTATGGCCAGC, TRPV3-C619S-Fw and GCTGGCCATAgGAACTGGAGTCC, TRPV3-C619S-Rv respectively. All cDNA constructs were confirmed by DNA-sequencing and used to make cRNAs. Oocytes expressing mTRPV3-C619S and mTRPV3-C612S mutant channels were challenged with 2-APB (1 mM), camphor (10 mM) and dihydrocarveol (10 mM) either at -40 mV or +40 mV holding potentials in voltage-clamp experiments. Responses of both mutants to 2-APB were similar to wild-type mTRPV3. Interestingly, responses to camphor were totally lost in mTRPV3-C619S mutant, while responses to dihydrocarveol remained intact. In contrast mTRPV3-C612S displayed slightly altered (16±2 % reduction) phenotype with respect to camphor sensitivity. It is concluded that pore-region cysteines play critical role in camphor sensitivity of TRPV3 ion channels.

Comparison of effects of extract of Myristica fragrans and verapamil on the volume and acidity of carbachol induced gastric secretion in fasting rabbits.

BACKGROUND: Peptic ulcer is mostly produced due to the over production of gastric acid. This study was undertaken to find out the effects of extract from Myristica fragrans which contains documented natural Calcium channel blocker and Verapamil on volume and acidity of Carbacol induced gastric section. Their effects were also compared to find out any difference in their efficacy. METHODS: Thirty rabbits of local breed, weighing 1-1.5 kg were used. The animals were kept on fasting for 48 hours, after which the pylorus of each animal was ligated. Verapamil 10 mg/kg, Myristica fragrans 500 mg/kg and Carbachol 600 microg/kg body weight were administered intraperitoneally. RESULTS: It was found that extract from Myristica fragrans reduced the volume, free and total acidity of gastric secretion, which were statistically highly significant when compared with Carbachol (P<0.001). Verapamil had also the same effects. When the difference of mean for verapamil was compared with that of extract, all these differences were found statistically non significant indicating that extract has similar effect as that of Verapamil on all parameters included in study. CONCLUSION: The effect of Myristica fragrans is similar to Verapamil and therefore it can be used effectively in the treatment of peptic ulcer and all other conditions that require calcium channel blockers for the treatment of these disorders.