Hepatoprotective Constituents of Macrocybe gigantea (Agaricomycetes) from India.

Non-alcoholic fatty liver disease (NAFLD) is one of the most frequent, chronic liver diseases worldwide and currently has no specific therapy. Our previous study indicated the anti-NAFLD effect of Macrocybe gigantea (Massee) Pegler & Lodge in high-fat diet-fed animals. This study aimed to isolate and identify the active hepatoprotective constituents from M. gigantea using fatty acid induced steatotic HepG2 cells as in vitro model. The effect of the test materials on the viability of HepG2 cells was analyzed using MTT assay. The HepG2 cells were treated with a mixture of palmitate-oleate to induce steatosis; after 24 h of treatment with the test materials, the intracellular lipid content was estimated using Oil Red O staining. The levels of transaminases were also estimated in the spent media. Bioassay-guided isolation of hepatoprotective constituents from M. gigantea yielded two compounds viz., ergosterol and linoleic acid; their structures were confirmed using spectroscopic data. Among these two compounds, ergosterol significantly lowered the levels of intracellular triglyceride content of fatty acid induced HepG2 cells; it also lowered the leakage of transaminases. The reductions caused by linoleic acid were not statistically significant at the tested concentrations. Detailed investigations on efficacy and safety of these compounds and M. gigantea might yield some useful leads for the management of NAFLD.

Screening of Weight-Loss Herbal Products for Synthetic Anti-Obesity Adulterants: A Target-Oriented Analysis by Liquid Chromatography-Tandem Mass Spectrometry.

BACKGROUND: The medical and social impact of being overweight, as well as the difficulty in making long-term changes in diet, physical activity, and pitfalls associated with allopathic management, lure obese individuals to over-the-counter weight-loss herbal products (WHPs). However, the safety of herbals is questionable, as reports from all over the world suggest adulteration with synthetic weight loss agents. Objective: The present study was carried out to develop a rapid and sensitive method to screen potential weight loss adulterants in WHPs. Methodology: We procured 33 licensed brands of WHPs that are available in Puducherry (India) and through web stores. Drug-free spiked herbal matrices were used for internal proficiency testing. Spiked herbal matrices and samples were extracted and centrifuged. The supernatant was collected and subjected to liquid chromatography-tandem mass spectrometry analyses. Isocratic elution was carried out by the mobile phase of methanol: 0.1% v/v formic acid (70%:30% v/v) through a C18 column. Analytes were monitored in electrospray ionization positive and negative modes, and an multiple reaction monitoring (MRM) scan was used. Results: Run time was 12 min. Intra- and inter-day precision was < 15% and recoveries were in the range of 99% to 115%. Limit of quantification (LOQ) of the analytes were in the range of 0.1 to 0.3 ng/ml. Conclusion: We developed a sensitive and rapid validated method for the detection of potential adulterants in WHPs. None of the WHPs analyzed were found to contain adulterants. The proposed method will be useful for routine monitoring. WHPs analyzed in this study were not adulterated indicating that WHPs from licensed sources are reliable.

Screening of herbal medicines for potential allopathic antidiabetic adulterants: An analytical study.

BACKGROUND: There are several reports worldwide on adulteration of herbal medicines (HMs) with allopathic drugs. In India, only a few studies have reported adulteration of HMs with antidiabetics and there are no systematic studies. AIMS: To develop a rapid and validated method for detection of allopathic antidiabetic adulterants and to explore the extent of adulteration in HMs sold in South India. MATERIALS AND METHODS: Standards and solvents were purchased from Sigma-Aldrich. Different brands of antidiabetic HM samples with manufacturing licenses were procured from dispensaries. Spiked drug free psyllium husk as solid and flask seed oil as liquid herbal matrices were used for method development. The spiked matrices with different concentrations were extracted with methanol and subjected to centrifugation. The supernatant was collected and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Isocratic elution was carried on a C18 column with 0.1% (v/v) formic acid:methanol (3:7, v/v) as a mobile phase. All drugs were monitored for two ion products in positive electrospray ionization mode using multiple reaction monitoring scans. RESULTS: The retention time was 9 min. Limit of detection is 10 Pictograms (pg) for all analytes except for metformin, which was 370 pg. Recoveries of analytes range from 96% to 117%. Forty different brands of antidiabetic HMs were analyzed. Adulterant peaks were not observed in the mass chromatograms of HMs. CONCLUSIONS: A single-run method was developed by LC-MS/MS for the detection of proposed antidiabetics in HMs from licensed manufacturing units and online sold HMs across herbal dispensaries in Puducherry union territory, India. None of the HMs was found to be adulterated with proposed allopathic antidiabetic adulterants.

Causes, consequences, and policy responses to the migration of health workers: key findings from India.

BACKGROUND: This study sought to better understand the drivers of skilled health professional migration, its consequences, and the various strategies countries have employed to mitigate its negative impacts. The study was conducted in four countries-Jamaica, India, the Philippines, and South Africa-that have historically been "sources" of health workers migrating to other countries. The aim of this paper is to present the findings from the Indian portion of the study. METHODS: Data were collected using surveys of Indian generalist and specialist physicians, nurses, midwives, dentists, pharmacists, dieticians, and other allied health therapists. We also conducted structured interviews with key stakeholders representing government ministries, professional associations, regional health authorities, health care facilities, and educational institutions. Quantitative data were analyzed using descriptive statistics and regression models. Qualitative data were analyzed thematically. RESULTS: Shortages of health workers are evident in certain parts of India and in certain specialty areas, but the degree and nature of such shortages are difficult to determine due to the lack of evidence and health information. The relationship of such shortages to international migration is not clear. Policy responses to health worker migration are also similarly embedded in wider processes aimed at health workforce management, but overall, there is no clear policy agenda to manage health worker migration. Decision-makers in India present conflicting options about the need or desirability of curtailing migration. CONCLUSIONS: Consequences of health work migration on the Indian health care system are not easily discernable from other compounding factors. Research suggests that shortages of skilled health workers in India must be examined in relation to domestic policies on training, recruitment, and retention rather than viewed as a direct consequence of the international migration of health workers.

Evaluating the Anticancer Potential of Ethanolic Gall Extract of Terminalia chebula (Gaertn.) Retz. (Combretaceae).

UNLABELLED: Plants have been an important source for discovery of anticancer compounds. With the current decline in the number of new molecular entities from the pharmaceutical industry, novel anticancer agents are being sought from traditional medicines; therefore the anticancer efficacy of many plants that are used in traditional medicine is yet to be verified. The objective of the study was to evaluate the cytotoxic potential of ethanolic leaf gall extract of Terminalia chebula are evaluated against buffalo rat liver 3A, MCF-7 (Human mammary gland adenocarcinoma) and A-549 (Human lung cancer) cell lines. The cytotoxic effect of the ethanolic extract was evaluated by MTT assay. The extract was potent and effective in inducing cytotoxic effects in all the cell lines with an IC50 value of 305.18 ± 1.7 µg/mL, 643.13 ± 4.2 µg/mL, and 208.16 ± 3.7 µ/mL, respectively. The extract was more effective against A549 cell lines when compared to others. The presences of phenolics, triterpenoids, and flavonoids were identified in the extract. The extract showed total phenolic and flavonoid content of 478 ± 2.2 mg of gallic acid equivalent/g d.w and 538 ± 1.4 mg of quercetinequivalent/g d.w, respectively. This higher content of total phenolics and flavonoids found in the ethanolic extract was directly associated to higher cytotoxicity activity. CONCLUSION: The ethanolic leaf gall extract of T. chebula showed effective cytotoxic activities; which might be attributed to the phenolics/flavonoids present in higher concentration. Future work will be interesting to know the chemical composition of the extract and also better understand the mechanism of action of the constituents present in the extract to develop it as drug for therapeutic application. SUMMARY: The present investigation establishes the anticancer activities of T. chebula leaf gall extracts on BRL3A, MCF-7, and A-549 cells. Presumably, these activities could be attributed in part to the phenolics/flavanoids features of the extract that has been demonstrated to act as cytotoxic agents. The experimental evidence obtained in the laboratory model could provide a rationale for the traditional use of plant as a source of easily available effective anticancer agents to the people, particularly in developing countries.

Antidiarrhoeal efficacy of Mangifera indica seed kernel on Swiss albino mice.

OBJECTIVE: To examine the antidiarrhoeal activity of alcoholic and aqueous seed kernel extract of Mangifera indica (M. indica) on castor oil-induced diarrhoeal activity in Swiss albino mice. METHODS: Mango seed kernels were processed and extracted using alcohol and water. Antidiarrhoeal activity of the extracts were assessed using intestinal motility and faecal score methods. RESULTS: Aqueous and alcoholic extracts of M. indica significantly reduced intestinal motility and faecal score in Swiss albino mice. CONCLUSIONS: The present study shows the traditional claim on the use of M. indica seed kernel for treating diarrhoea in Southern parts of India.

Antifungal activity of Ocimum sanctum Linn. (Lamiaceae) on clinically isolated dermatophytic fungi.

OBJECTIVE: To assess antifungal activity of Ocimum sanctum leaves against dermatophytic fungi. METHODS: Antifungal activity of Ocimum sanctum leaves was measured by 38 A NCCLS method. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of various extracts and fractions of Ocimum sanctum leaves were also determined. RESULTS: Ocimum sanctum leaves possessed antifungal activity against clinically isolated dermatophytes at the concentration of 200 µg/mL. MIC and MFC were high with water fraction (200 µg/mL) against dermatophytic fungi used. CONCLUSIONS: Ocimum sanctum has antifungal activity, and the leaf extracts may be a useful source for dermatophytic infections.

Anti-enteric bacterial activity and phytochemical analysis of the seed kernel extract of Mangifera indica Linnaeus against Shigella dysenteriae (Shiga, corrig.) Castellani and Chalmers.

OBJECTIVE: To evaluate the phytochemical and anti-bacterial efficacy of the seed kernel extract of Mangifera indica (M. indica) against the enteropathogen, Shigella dysenteriae (S. dysenteriae), isolated from the diarrhoeal stool specimens. METHODS: The preliminary phytochemical screening was performed by the standard methods as described by Harborne. Cold extraction method was employed to extract the bioactive compounds from mango seed kernel. Disc diffusion method was adopted to screen antibacterial activity. Minimum inhibitory concentration (MIC) was evaluated by agar dilution method. The crude extracts were partially purified by thin layer chromatography (TLC) and the fractions were analyzed by high performance thin layer chromatography (HPTLC) to identify the bioactive compounds. RESULTS: Phytochemical scrutiny of M. indica indicated the presence of phytochemical constituents such as alkaloids, gums, flavanoids, phenols, saponins, steroids, tannins and xanthoproteins. Antibacterial activity was observed in two crude extracts and various fractions viz. hexane, benzene, chloroform, methanol and water. MIC of methanol fraction was found to be (95±11.8) µg/mL. MIC of other fractions ranged from 130-380 µg/mL. CONCLUSIONS: The present study confirmed that each crude extracts and fractions of M. indica have significant antimicrobial activity against the isolated pathogen S. dysenteriae. The antibacterial activity may be due to the phytochemical constituents of the mango seed kernel. The phytochemical tannin could be the reason for its antibacterial activity.

Antifungal activity of Aegle marmelos (L.) Correa (Rutaceae) leaf extract on dermatophytes.

OBJECTIVE: To evaluate the in vitro antifungal activity of Aegle marmelos leaf extracts and fractions on the clinical isolates of dermatophytic fungi like Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, Microsporum gypseum and Epidermophyton floccosum. METHODS: The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of various extracts and fractions of the leaves of Aegle marmelos were measured using method of National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: Aegle marmelos leaf extracts and fractions were found to have fungicidal activity against various clinical isolates of dermatophytic fungi. The MIC and MFC was found to be high in water and ethyl alcohol extracts and methanol fractions (200µg/mL) against dermatophytic fungi studied. CONCLUSIONS: Aegle marmelos leaf extracts significantly inhibites the growth of all dermatophytic fungi studied. If this activity is confirmed by in vivo studies and if the compound is isolated and identified, it could be a remedy for dermatophytosis.

An improved pollen collection and cryopreservation method for highly recalcitrant tropical fruit species of mango (Mangifera indica L.) and litchi (Litchi chinensis Sonn.).

An improved method for pollen collection from freshly dehiscing anthers of mango (Mangifera indica L.) and litchi (Litchi chinensis Sonn.) using the organic solvent cyclohexane has been devised. Using this method pollen quantity sufficient for large scale pollinations could be collected and stored for future use. Transport of pollen in viable conditions over long distances, from site of collection (field genebank) to cryolab was successfully devised for both these fruit species. Cryopreservation was successfully applied to achieve long-term pollen storage over periods of up to four years. Pollen viability was tested using in vitro germination, the fluorochromatic reaction (FCR) method and by fruit set following field pollination. On retesting, four year cryostored pollen of different mango and litchi varieties showed high percentage viability as good as fresh control pollens. Pollens of more than 180 cultivars of mango and 19 cultivars of litchi have been stored in the cryogenebank using the technology developed, thus facilitating breeding programmes over the long-term.

Nimbolide and isonimbolide.

Nimbolide 1, a potent molecule of biological significance, was isolated. Attempts were made to cleave the ether linkage in nimbolide using boron trifluoride etherate in the presence of tetrabutyl ammonium bromide so as to generate a ring-opened structure akin to azadirachtins, which are known to possess excellent antifeedant properties. However, a novel rearranged product was envisaged during the course of the reaction, which was determined as isonimbolide 2-a structural isomer of nimbolide through spectroscopic methods.

Determining hierarchical functional networks from auditory stimuli fMRI.

We determined connectivity of the human brain using functional magnetic resonance imaging (fMRI) while subjects experienced auditory stimuli in a 2-by-2 factorial design. The two factors in this study were "speaker" (same or different speaker) and "sentence" (same or different sentences). Connectivity studies allow us to ask how spatially remote brain regions are neurophysiologically related given these stimuli. In the context of this study, we examined how the "speaker" effect and "sentence" effect influenced these relationships. We applied a Bayesian connectivity method that determines hierarchical functional networks of functionally connected brain regions. Hierarchy in these functional networks is determined by conditional probabilities of elevated activity. For example, a brain region that becomes active a superset of the time of another region is considered ascendant to that brain region in the hierarchical network. For each factor level, we found a baseline functional network connecting the primary auditory cortex (Brodmann's Area [BA] 41) with the BA 42 and BA 22 of the superior temporal gyrus (STG). We also found a baseline functional network that includes Wernicke's Area (BA 22 posterior), STG, and BA 44 for each factor level. However, we additionally observed a strong ascendant connection from BA 41 to the posterior cingulate (BA 30) and Broca's Area and a stronger connection from Wernicke's Area to STG and the posterior cingulate while passively listening to different sentences rather than the same sentence repeatedly. Finally, our results revealed no significant "speaker" effect or interaction between "speaker" and "sentence."

Antifeedant neo-clerodanes from Teucrium tomentosum Heyne. (Labiatae).

From the acetone extract of Teucrium tomentosum, a new antifeedant neo-clerodane diterpenoid teuctosin (1) was isolated along with teuflin (2), teucrin-H(2) (3), 6beta-hydroxyteuscordin (4), 6beta-acetylteuscordin (5) and montanin-D (6). The structure of the new compound was elucidated comprehensively using 1D and 2D NMR methods and confirmed by X-ray crystallography. All the compounds showed effective antifeedancy against Plutella xylostella and Spodoptera litura at 10 mug/cm(2) of leaf area.

8-(2-bromo-3-methoxy-3-methylbutyl)-7-methoxycoumarin.

The title compound, C(16)H(19)BrO(4), is a derivative of osthol, isolated from the seeds of Imperatoria Osthruthium. The structure was solved in space group P-1, with two molecules in the asymmetric unit, and was refined to a final R factor of 0.064. The two molecules in the asymmetric unit differ in the orientation of their brominated substituent group. The benzopyran ring displays aromatic character. The packing of the molecules in the lattice is mainly due to C-H...O hydrogen bonds.

Antifertility activity of Derris brevipes variety coriacea.

Traditional physicians in and around Kotagiri village near Ootacamund, use a mixture of powdered roots of Cassia occidentalis, Derris brevipes variety coriacea and Justicia simplex to control female fertility. A mixture of powdered roots of these three plants, powdered root of Derris brevipes variety coriacea and its ethanolic extract were screened for antifertility activity in proven fertile female rats at 200 and 600 mg/kg body weight, respectively and given orally on D(1-7) of pregnancy. Both doses of the root powder of Derris brevipes variety coriacea showed 50% anti-implantation activity and also a significant reduction in the number of litters born. The ethanolic extract exhibited 40% anti-implantation activity when given orally at 600 mg/kg body weight. The rats, which continued their pregnancy, did not deliver any litters after their full term. Hence, the combined antifertility (anti-implantation and abortifacient) activity of the ethanolic extract was 100%. The results suggest that the ethanolic extract possesses more abortifacient type effect than the anti-implantation activity. The ethanolic extract also exhibited weak estrogenic activity when given alone and tested in immature ovariectomised female albino rats. But, when given along with ethinyl estradiol, it exhibited slight antiestrogenic activity. Histological and biochemical estimations were carried out to confirm this.

Pharmacognostical Evaluation of Caesalpinia sappan Heartwood.

Caesalpinia span heartwood is used in folklore in India. The present work attempts to summarize the pharmacognostical characters of the heartwood of this plant. Ash and extractive values, phytochemical tests, HPTLC and fluorescence analysis were carried out. The present work contributes for the standardization of this medicinal heat wood.

Ethnobiology of the Nilgiri hills, India.

The Nilgiri is a popular massif towering high in the Western Ghats in South India with an altitude of 2623 m. Nature has been magnanimous in bestowing Nilgiri district with rich evergreen temperate to tropical forests. A high degree of biodiversity, marked by varied flora and fauna of good therapeutic potential as well as the varied number of indigenous groups of people in this area, makes it very popular among herbalists. The district has six anthropologically well defined ethnic groups namely Todas, Kotas, Kurumbas, Irulas, Paniyas and Kattunayakas living here possibly since 1200 B.C. The present review highlights the ethnobiological profile of six indigenous populations and their dependence on ambient flora and fauna for traditional health care needs. It has been observed that about 2700 therapeutically potent plant species are available in this hill station of which almost all have come from local medicine. Some have been explored scientifically. However, about 150 plant species are still to be explored for their therapeutic potential. The ethnography, phytochemical and therapeutic uses as well as the anthropological perspectives of the local medicines have been discussed in this review.

Expression pattern in brain of TASK-1, TASK-3, and a tandem pore domain K(+) channel subunit, TASK-5, associated with the central auditory nervous system.

TWIK-related acid-sensitive K(+) (TASK) channels contribute to setting the resting potential of mammalian neurons and have recently been defined as molecular targets for extracellular protons and volatile anesthetics. We have isolated a novel member of this subfamily, hTASK-5, from a human genomic library and mapped it to chromosomal region 20q12-20q13. hTASK-5 did not functionally express in Xenopus oocytes, whereas chimeric TASK-5/TASK-3 constructs containing the region between M1 and M3 of TASK-3 produced K(+) selective currents. To better correlate TASK subunits with native K(+) currents in neurons the precise cellular distribution of all TASK family members was elucidated in rat brain. A comprehensive in situ hybridization analysis revealed that both TASK-1 and TASK-3 transcripts are most strongly expressed in many neurons likely to be cholinergic, serotonergic, or noradrenergic. In contrast, TASK-5 expression is found in olfactory bulb mitral cells and Purkinje cells, but predominantly associated with the central auditory pathway. Thus, TASK-5 K(+) channels, possibly in conjunction with auxiliary proteins, may play a role in the transmission of temporal information in the auditory system.

THIK-1 and THIK-2, a novel subfamily of tandem pore domain K+ channels.

Two cDNAs encoding novel K(+) channels, THIK-1 and THIK-2 (tandem pore domain halothane inhibited K(+) channel), were isolated from rat brain. The proteins of 405 and 430 amino acids were 58% identical to each other. Homology analysis showed that the novel channels form a separate subfamily among tandem pore domain K(+) channels. The genes of the human orthologs were identified as human genomic data base entries. They possess one intron each and were assigned to chromosomal region 14q24.1-14q24.3 (human (h) THIK-1) and 2p22-2p21 (hTHIK-2). In rat (r), THIK-1 (rTHIK-1) is expressed ubiquitously; rTHIK-2 expression was found in several tissues including brain and kidney. In situ hybridization of brain slices showed that rTHIK-2 is strongly expressed in most brain regions, whereas rTHIK-1 expression is more restricted. Heterologous expression of rTHIK-1 in Xenopus oocytes revealed a K(+) channel displaying weak inward rectification in symmetrical K(+) solution. The current was enhanced by arachidonic acid and inhibited by halothane. rTHIK-2 did not functionally express. Confocal microscopy of oocytes injected with green fluorescent protein-tagged rTHIK-1 or rTHIK-2 showed that both channel subunits are targeted to the outer membrane. However, coinjection of rTHIK-2 did not affect the currents induced by rTHIK-1, indicating that the two channel subunits do not form heteromers.

Photooxidation of cedrelone, a tetranortriterpenoid from Toona ciliata.

Cedrelone, a tetranortriterpenoid on photolysis by UV light yields a true photooxidation product 3 [14 beta,15beta,22beta,23beta-diepoxy-6-hydroxy-1,5, 20(22)-meliatriene-2,7,21-trione] whose structure is well established by NMR studies and confirmed by X-ray crystallography, along with product 4 [14 beta,15beta-epoxy-6,23-dihydroxy-1,5,20(22)-meliatriene-2,7, 21-trione]. Addition of rose bengal increases the rate of photooxidation whereas DABCO decreases rate of photolysis proving the involvement of singlet oxygen in the photooxygenation. Both the photoproducts exhibited antifeedant activity.