Harnessing the Potential of Roots of Traditional Power Plant: Ocimum.

Genus Ocimum of Labiatae is well known in all traditional medicinal systems like Ayurveda, Unani, Siddha, and Homeopathy. The pharmaceutical activities of different species of Ocimum attributed to all plant parts. Roots are the most significant vital organ of the plant, as they absorb water and nutrients from soil and transport to aerial parts of the plants. Roots of Ocimum were found helpful with free-radical scavenging activity to improve physical and mental strength as well as to treat diabetes, malaria, and liver problems. Antibacterial activity of Ocimum roots and its main component, rosmarinic acid, is very beneficial to protect against several human pathogens, including bacteria and viruses. Being so important in every way, roots of Ocimum need healthy rhizosphere. Bacteria, fungi, nematodes, types of soil, fungicide, pesticides, salt, radioactive elements, as well as heavy metal contaminations, affect roots and overall growth of Ocimum in positive or negative ways. Each component of rhizosphere (natural, treatment or contamination) affects the roots, which highlights current ecological scenario to discover biosafe and more productive approaches. For such prestigious organ of Ocimum, development of in vitro root cultures and hairy root cultures assists to reduce the efforts and timing of the traditional cultivation process along with elimination of negative factors in rhizosphere. Different strains of Agrobacterium rhizogenes, various media compositions, as well as discrete treatments, like elicitors, on nonidentical species or cultivars of Ocimum boost the root induction, biomass, and accumulation of phytoceuticals differently. Hairy roots and in vitro roots of Ocimum accumulate higher quantity of therapeutic metabolites. These metabolites include several phenolics (like rosmarinic acid, 3-hydroxybenzoic acid, m-coumaric acid, p-coumaric acid, caffeic acid, ferulic acid, vanillic acid, chicoric acid, and lithospermic acid), triterpenes (such as betulinic acid, 3-epimaslinic acid, alphitolic acid, euscaphic acids, oleanolic acid, and ursolic acid) as well as flavonoids (flavones, flavonols, and dihydroflavonols). This review highlights pharmaceutical applications of Ocimum roots, a great deal of rhizosphere components and in vitro culturing techniques to enhance biomass as well as chief phytoceuticals.

Efficacy of autologous stem cells for bone regeneration during endosseous dental implants insertion - A systematic review of human studies.

Availability of adequate quantity and quality of bone is prerequisite for longevity and survival of endosseous dental implants. Most of the clinicians face with the problem of lack of bone due to long-standing edentulism during this treatment modality. Conventional therapies with the use of various types of bone grafts and membranes have provided clinicians with unpredictable and compromised results. Cell-based therapies utilizing undifferentiated cells, that have the potential to differentiate into various cell types including osteoblastic lineages, have demonstrated through various previously conducted in-vitro and animal studies, a successful formation of bone in a predictable manner. Thus the main objective of this review was to evaluate the effectiveness of these therapies when applied on human subjects. A search was carried out in MEDLINE (via PubMed) and Cochrane CENTRAL databases for completed randomized and non-randomised clinical trials utilizing stem cell-based therapies with histologic and radiographic analysis written in English up to January 2019. This search of the literature yielded 10 studies meeting the inclusion and exclusion criteria. In all these studies, stem cells were primarily used to achieve bone augmentation during insertion of endosseous dental implants. Results of these therapies conducted on human subjects have shown a positive impact on bone regeneration, in particular, therapies utilizing bone marrow and adipose tissue derived stem cells. But the clinicians need to examine the efficacy, safety, feasibility of these therapies while treating large size defects or planning for shorter healing period and early loading of dental implants.

Modes of Calcium Regulation in Ischemic Neuron.

Calcium (Ca2+) dysregulation is a major catalytic event. Ca2+ dysregulation leads to neuronal cell death and brain damage result in cerebral ischemia. Neurons are unable in maintaining calcium homeostasis. Ca2+ homeostasis imbalance results in increased calcium influx and impaired calcium extrusion across the plasma membrane. Ca2+ dysregulation is mediated by different cellular and biochemical mechanism, which leads to neuronal loss resulting stroke/cerebral ischemia. A better understanding of the Ca2+ dysregulation might help in the development of new treatments in order to reduce ischemic brain injury. An optimal concentration of Ca2+ does not lead to neurotoxicity in the ischemic neuron. Intracellular Ca2+ act as a trigger for acute neurotoxicity and this cause induction of long-lasting processes leading to necrotic and/or apoptotic post-ischemic delayed neuronal death or of compensatory, neuroprotective mechanisms has increased considerably. Moreover, routes of ischemic Ca2+ influx to neurons, involvement of intracellular Ca2+ stores and Ca2+ buffers, spatial and temporal relations between ischemia-induced increases in intracellular Ca2+ concentration and neurotoxicity will further increase our understanding about underlying mechanism and they can act as a target for the development of drugs. Here, in our article we are trying to provide a brief overview of various Ca2+ influx pathways involve in ischemic neuron and how ischemic neuron attempts to counterbalance this calcium overload.

Antimicrobial effects of various platelet rich concentrates-vibes from in-vitro studies-a systematic review.

AIM: The aim of the current review was to outline the existing information related to antimicrobial properties of various platelet concentrates, as experimented in various in-vitro studies. BACKGROUND: One of the most interesting recent landmarks in the field of biological therapy has been the discovery that platelets, in addition to being capable of releasing hundreds of proteins and growth factors, can also release immunomodulatory agents with antimicrobial activity. Several international research groups have reported antimicrobial activities in both human platelets and other types of platelet rich plasma preparations. REVIEW RESULT: This review was carried-out pursuing a systematic approach. An electronic search was conducted on MEDLINE and GOOGLE SCHOLAR databases using suitable search terminologies. It included preclinical studies which assessed the antimicrobial activity of Autologous Platelet Concentrates(APC).Ten in-vitro studies and one animal study, which investigated APC effects on various microorganisms, were included. Almost in all the included in-vitro studies, it was found that complete breakdown of microbial load could not be achieved by any of the APC preparations but there occurred a reduction in the growth of microorganisms. Thus APCs displayed a bacteriostatic rather than bacteriocidal activity.The only animal study included in this review which had both in-vitro and in-vivo evaluation, also showed reduction of infection caused by different microorganisms. CONCLUSION: Although the precise mechanism of synergy with microbial pathogens needs further validation, platelet concentrates proved to have antimicrobial properties.

Withania somnifera: Advances and Implementation of Molecular and Tissue Culture Techniques to Enhance Its Application.

Withania somnifera, commonly known as Ashwagandha an important medicinal plant largely used in Ayurvedic and indigenous medicine for over 3,000 years. Being a medicinal plant, dried powder, crude extract as well as purified metabolies of the plant has shown promising therapeutic properties. Withanolides are the principal metabolites, responsible for the medicinal properties of the plant. Availability and amount of particular withanolides differ with tissue type and chemotype and its importance leads to identification characterization of several genes/ enzymes related to withanolide biosynthetic pathway. The modulation in withanolides can be achieved by controlling the environmental conditions like, different tissue culture techniques, altered media compositions, use of elicitors, etc. Among all the in vitro techniques, hairy root culture proved its importance at industrial scale, which also gets benefits due to more accumulation (amount and number) of withanolides in roots tissues of W. somnifera. Use of media compostion and elicitors further enhances the amount of withanolides in hairy roots. Another important modern day technique used for accumulation of desired secondary metabolites is modulating the gene expression by altering environmental conditions (use of different media composition, elicitors, etc.) or through genetic enginnering. Knowing the significance of the gene and the key enzymatic step of the pathway, modulation in withanolide contents can be achieved upto required amount in therapeutic industry. To accomplish maximum productivity through genetic enginnering different means of Withania transformation methods have been developed to obtain maximum transformation efficiency. These standardized transformation procedues have been used to overexpress/silence desired gene in W. somnifera to understand the outcome and succeed with enhanced metabolic production for the ultimate benefit of human race.

ROS-dependent genotoxicity, cell cycle perturbations and apoptosis in mouse bone marrow cells exposed to formulated mixture of cypermethrin and chlorpyrifos.

Swiss albino mice were exposed to formulated cypermethrin (CMR) and/or or chlorpyrifos (CPF) through oral gavages for 60 days. Test doses of CMR (0.69, 1.38 or 2.76mg/kg/day) or CPF (0.5, 1.0 or 2.0mg/kg/day) or CMR + CPF (0.69 + 0.5, 1.38 + 1.0 or 2.76 + 2.0mg/kg/day) were based on the acute oral median lethal doses of CMR or CPF. Chromosome aberrations (CA), micronucleus (MN) induction, cell cycle perturbations, apoptosis and reactive oxygen species (ROS) generation were analysed in bone marrow cells. To explore the involvement of ROS induction, HaCat cells were exposed in vitro to arbitrary concentrations of CMR and/or CPF. Exposure of CMR (2.76mg/kg/day) induced significant inhibition of mitotic index. Significant (P < 0.01) frequencies of CA and MN were observed with the CMR at 1.38mg/kg/day, whereas CPF or its mixture CMR + CPF showed at highest doses. Chromosome/chromatid breaks and fragments were found to be major aberrations in all the treatment groups. Highest doses of CMR or CMR + CPF revealed significant (P < 0.01 or 0.001) elevation of G0/G1 peak, while CPF-exposed cells revealed significant (P < 0.01) declined in G1 phase. Decline in S phase was observed with highest dose of CMR only. Apoptosis induction measured by gating cell population beside G1 peak showed 3- to 4-fold increase in apoptotic cells in CPF-exposed mice as compared to control or CMR or CMR + CPF-treated mice. Further, all the treatment groups in vivo as well as in vitro revealed significant generation of ROS in comparison with the control group. Present results, together with the earlier reports, which substantiate ROS generation may be major cause of genotoxicity, cell cycle perturbations and apoptosis, nonetheless co-exposure of low doses of CMR and CPF mixture does not potentiate genotoxicity.

Comparative interactions of withanolides and sterols with two members of sterol glycosyltransferases from Withania somnifera.

BACKGROUND: Sterol glycosyltransferases (SGTs) are ubiquitous but one of the most diverse group of enzymes of glycosyltransferases family. Members of this family modulate physical and chemical properties of secondary plant products important for various physiological processes. The role of SGTs has been demonstrated in the biosynthesis of pharmaceutically important molecules of medicinal plants like Withania somnifera. RESULTS: Analysis suggested conserved behaviour and high similarity in active sites of WsSGTs with other plant GTs. Substrate specificity of WsSGTs were analysed through docking performance of WsSGTs with different substrates (sterols and withanolides). Best docking results of WsSGTL1 in the form of stable enzyme-substrate complex having lowest binding energies were obtained with brassicasterol, transandrosteron and WsSGTL4 with solasodine, stigmasterol and 24-methylene cholesterol. CONCLUSION: This study reveals topological characters and conserved nature of two SGTs from W. somnifera (WsSGTs) i.e. WsSGTL1 and WsSGTL4. However, besides being ubiquitous in nature and with broad substrate specificity, difference between WsSGTL1 and WsSGTL4 is briefly described by difference in stability (binding energy) of enzyme-substrate complexes through comparative docking.

WsSGTL1 gene from Withania somnifera, modulates glycosylation profile, antioxidant system and confers biotic and salt stress tolerance in transgenic tobacco.

Glycosylation of sterols, catalysed by sterol glycosyltransferases (SGTs), improves the sterol solubility, chemical stability and compartmentalization, and helps plants to adapt to environmental changes. The SGTs in medicinal plants are of particular interest for their role in the biosynthesis of pharmacologically active substances. WsSGTL1, a SGT isolated from Withania somnifera, was expressed and functionally characterized in transgenic tobacco plants. Transgenic WsSGTL1-Nt lines showed an adaptive mechanism through demonstrating late germination, stunted growth, yellowish-green leaves and enhanced antioxidant system. The reduced chlorophyll content and chlorophyll fluorescence with decreased photosynthetic parameters were observed in WsSGTL1-Nt plants. These changes could be due to the enhanced glycosylation by WsSGTL1, as no modulation in chlorophyll biogenesis-related genes was observed in transgenic lines as compared to wildtype (WT) plants. Enhanced accumulation of main sterols like, campesterol, stigmasterol and sitosterol in glycosylated form was observed in WsSGTL1-Nt plants. Apart from these, other secondary metabolites related to plant's antioxidant system along with activities of antioxidant enzymes (SOD, CAT; two to fourfold) were enhanced in WsSGTL1-Nt as compared to WT. WsSGTL1-Nt plants showed significant resistance towards Spodoptera litura (biotic stress) with up to 27 % reduced larval weight as well as salt stress (abiotic stress) with improved survival capacity of leaf discs. The present study demonstrates that higher glycosylation of sterols and enhanced antioxidant system caused by expression of WsSGTL1 gene confers specific functions in plants to adapt under different environmental challenges.

Overexpression of WsSGTL1 gene of Withania somnifera enhances salt tolerance, heat tolerance and cold acclimation ability in transgenic Arabidopsis plants.

BACKGROUND: Sterol glycosyltrnasferases (SGT) are enzymes that glycosylate sterols which play important role in plant adaptation to stress and are medicinally important in plants like Withania somnifera. The present study aims to find the role of WsSGTL1 which is a sterol glycosyltransferase from W. somnifera, in plant's adaptation to abiotic stress. METHODOLOGY: The WsSGTL1 gene was transformed in Arabidopsis thaliana through Agrobacterium mediated transformation, using the binary vector pBI121, by floral dip method. The phenotypic and physiological parameters like germination, root length, shoot weight, relative electrolyte conductivity, MDA content, SOD levels, relative electrolyte leakage and chlorophyll measurements were compared between transgenic and wild type Arabidopsis plants under different abiotic stresses--salt, heat and cold. Biochemical analysis was done by HPLC-TLC and radiolabelled enzyme assay. The promoter of the WsSGTL1 gene was cloned by using Genome Walker kit (Clontech, USA) and the 3D structures were predicted by using Discovery Studio Ver. 2.5. RESULTS: The WsSGTL1 transgenic plants were confirmed to be single copy by Southern and homozygous by segregation analysis. As compared to WT, the transgenic plants showed better germination, salt tolerance, heat and cold tolerance. The level of the transgene WsSGTL1 was elevated in heat, cold and salt stress along with other marker genes such as HSP70, HSP90, RD29, SOS3 and LEA4-5. Biochemical analysis showed the formation of sterol glycosides and increase in enzyme activity. When the promoter of WsSGTL1 gene was cloned from W. somnifera and sequenced, it contained stress responsive elements. Bioinformatics analysis of the 3D structure of the WsSGTL1 protein showed functional similarity with sterol glycosyltransferase AtSGT of A. thaliana. CONCLUSIONS: Transformation of WsSGTL1 gene in A. thaliana conferred abiotic stress tolerance. The promoter of the gene in W.somnifera was found to have stress responsive elements. The 3D structure showed functional similarity with sterol glycosyltransferases.

In vitro isolation and characterization of biolarvicidal compounds from micropropagated plants of Spilanthes acmella.

Spilanthes acmella (Family: Asteraceae) commonly known as "toothache plant" is known to possess strong insecticidal and larvicidal properties. Experiments have been conducted to isolate and characterise the biolarvicidal compounds from the flower head extract of micropropagated S. acmella plants employing various tools like FT-IR, TLC, CC, NMR. FT-IR spectroscopy of the crude hexane extract sample revealed the presence of amide (secondary metabolite) as functional group in S. acmella flower heads. The crude extract was separated into 85 fractions (100 ml each) through silica gel column chromatography using hexane-ethyl acetate mobile phase. All fractions were tested for their larvicidal activity against late III/early IV instar Anopheles stephensi larvae and fraction showing maximum bioefficacy against aforesaid larvae was further resolved into three separate bands on Preparative TLC plate, the respective R (f) values being (a) 0.18, (b) 0.23 and (c) 0.27. Based on Proton NMR spectrum of the eluted compounds and their comparison with published results, three different compounds were identified: N-isobutyl-2,6,8-decatrienamide (compound 1), undeca-2E,7Z,9E-trienoic acid isobutylamide (compound 2) from band a and (2E)-N-(2-methylbutyl)-2-undecene-8,10-diynamide (compound 3) from band b. The amount of the compounds obtained were 338 mg (compounds 1 and 2) and 188.4 mg (compound 3), respectively. This is the first report of biolarvicidal compounds isolation and characterisation from micropropagated S. acmella plants.

Agrobacterium tumefaciens-mediated transformation of Withania somnifera (L.) Dunal: an important medicinal plant.

This report describes Agrobacterium tumefaciens-mediated transformation of Withania somnifera--an important Indian medicinal plant. A. tumefaciens strain LBA4404, containing the binary vector pIG121Hm was used for transformation, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter. The leaf segments from two-and-a-half-month-old green house-grown seedlings were more efficient in transformation, as compared to those from the in vitro-grown shoots. Second expanded leaf from the shoot tip gave the highest transient transformation efficiency. Selection of transgenic shoots was done in the presence of 50 mg l(-1) kanamycin. Polymerase chain reaction analysis of T(0) transgenic plants showed the presence of gusA and nptII genes. The expression of these transgenes in T(1) progeny was confirmed by RT-PCR. The integration of gusA gene was confirmed by Southern blot analysis. The transformation efficiency was found to be 1.67%.

Emergency medical epidemiology in Assam, India.

BACKGROUND: Assam, with its capital in Dispur has one of the highest rates of infant and maternal mortality in India. Being under both tribal and hilly regions, it has lacked adequate healthcare and emergency services. We therefore aimed to conduct a cross-sectional survey of medical emergencies and identify various types of emergencies presenting to emergency departments, prior to launching emergency services across the state. MATERIALS AND METHODS: On a prospective basis and using a stratified random sampling design, all emergencies presenting to the three government hospitals in Guwahati, Assam, which handle 90% of all emergencies currently, were studied on specially designed datasheets in order to collect data. Emergency medical technicians (EMTs) were placed in the Casualty of the medical colleges and recorded all emergencies on the datasheet. The collected data was then analysed for stratification and mapping of emergencies. In addition, retrospective data for a period of 15 days was collected from the emergency case registers of all three hospitals and the adjoining district civil hospitals, in order to give a wider perspective of the nature of emergencies. RESULTS: A total of 2169 emergencies were recorded over a seven-day prospective and fifteen-day retrospective period. Guwahati Medical College Hospital attended to majority of emergencies (42%), which were mainly of the nature of pregnancies (22.7%), accidents (12.2%) or assaults (15.4%) and fever related. Maximum emergencies also presented from the border districts, and occurred among young males in the age group of 19-45 years. Males were also more prone to accidents and assaults, while females presented with pregnancies as emergencies. CONCLUSION: Potential emergency services need to target young pregnant females. Law and order needs to be also tightened in order to curb accidents and assaults among young males.

Alzheimer's and non-alzheimer's dementia: a critical review of pharmacological and nonpharmacological strategies.

OBJECTIVE: Dementia is an age-related progressive neurodegenerative disorder afflicting about 5% of the world's population, and it is expected to grow dramatically in the future keeping in view our ageing society. Currently available medications appear to be able to produce moderate symptomatic benefits but do not to stop disease progression. In this article, the management of the disorder, including the currently available drugs as well as psychosocial strategies, is discussed. METHODS: A computerized search on Pubmed from 1980 to 2006 was carried out and all articles evaluated and graded on NICE guidelines. RESULTS AND CONCLUSIONS: Currently evaluated and accepted medications only bring about a reduction in the deteriorating course. A combination of pharmacotherapy and psychosocial management is the need of the hour.

Strong larvicidal activity of three species of Spilanthes (Akarkara) against malaria (Anopheles stephensi Liston, Anopheles culicifacies, species C) and filaria vector (Culex quinquefasciatus Say).

A system for biocontrol of malaria and filarial mosquito vectors has been developed using herbal extracts of three Spilanthes species, S. acmella L.var oleraceae Clarke, S. calva L. and S. paniculata Wall ex DC. Cent percent mortalities was achieved against the late third/early fourth instar larvae of A. stephensi Liston, A. culicifacies species C and C. quinquefasciatus Say using crude hexane extract obtained from flower heads of Spilanthes spp. Of the three plant species, S. acmella extract proved to be the most effective in inducing complete lethality at minimum doses, the respective LC50 and LC90 values being 4.57 and 7.83 (A. stephensi), 0.87 and 1.92 (A. culicifacies) and 3.11 and 8.89 ppm (C. quinquefasciatus). This was followed by S. calva and S. paniculata extracts, respectively. This is the first report of achieving cent percent lethality against these mosquito larvae using minimal doses of plant extracts from this or any other plant species.