Finger millet bran supplementation alleviates obesity-induced oxidative stress, inflammation and gut microbial derangements in high-fat diet-fed mice.

Several epidemiological studies have shown that the consumption of finger millet (FM) alleviates diabetes-related complications. In the present study, the effect of finger millet whole grain (FM-WG) and bran (FM-BR) supplementation was evaluated in high-fat diet-fed LACA mice for 12 weeks. Mice were divided into four groups: control group fed a normal diet (10 % fat as energy); a group fed a high-fat diet; a group fed the same high-fat diet supplemented with FM-BR; a group fed the same high-fat diet supplemented with FM-WG. The inclusion of FM-BR at 10 % (w/w) in a high-fat diet had more beneficial effects than that of FM-WG. FM-BR supplementation prevented body weight gain, improved lipid profile and anti-inflammatory status, alleviated oxidative stress, regulated the expression levels of several obesity-related genes, increased the abundance of beneficial gut bacteria (Lactobacillus, Bifidobacteria and Roseburia) and suppressed the abundance of Enterobacter in caecal contents (P≤ 0·05). In conclusion, FM-BR supplementation could be an effective strategy for preventing high-fat diet-induced changes and developing FM-BR-enriched functional foods.

Microbial community structure at different fermentation stages of kutajarista, a herbal formulation.

Kutajarista is an Ayurvedic fermented herbal formulation prescribed for gastrointestinal disorders. This herbal formulation undergoes a gradual fermentative process and takes around 2 months for production. In this study, microbial composition at initial stages of fermentation of Kutajarista was assessed by culture independent 16S rRNA gene clone library approach. Physicochemical changes were also compared at these stages of fermentation. High performance liquid chromatography-mass spectrometry analysis showed that Gallic acid, Ellagic acid, and its derivatives were the major chemical constituents recovered in this process. At 0 day of fermentation, Lactobacillus sp., Acinetobacter sp., Alcaligenes sp., and Methylobacterium sp. were recovered, but were not detected at 8 day of fermentation. Initially, microbial diversity increased after 8 days of fermentation with 11 operational taxonomic units (OTUs), which further decreased to 3 OTUs at 30 day of fermentation. Aeromonas sp., Pseudomonas sp., and Klebsiella sp. dominated till 30 day of fermentation. Predominance of γ- Proteobacteria and presence of gallolyl derivatives at the saturation stage of fermentation implies tannin degrading potential of these microbes. This is the first study to highlight the microbial role in an Ayurvedic herbal product fermentation.

A new anxiolytic fatty acid from Aethusa cynapium.

The present investigation was carried out with a view to separate bioactive constituent from Aethusa cynapium. Bioactivity guided fractionation of the anxiolytic methanol extract has led to the isolation of a novel unsaturated fatty acid. Structure of the acid characterized by UV, IR, 1H NMR, C13 NMR, MS techniques was found to be trideca-7, 9, 11-trienoic acid. Antianxiety activity was confirmed using the mCPP-induced hypolocomotion test. This new fatty acid-trideca-7,9,11-trienoic acid, isolated from A. cynapium was found to be responsible for the antianxiety activity of the plant.

Apoptosis inducing activity of steroidal constituents from Solanum xanthocarpum and Asparagus racemosus.

A series of Sarsapogenin and Diosgenin derived steroidal constituents (1-12), isolated from Solanum xanthocarpum and Asparagus racemosus were screened for their ability to induce cell death and apoptosis of colon carcinoma cells. The carbohydrate moieties linked to the steroid backbones were found to strongly influence cytotoxic activity and cell death mode (apoptosis or necrosis). Compound 10, from A. racemosus was found to be a potent inducer of apoptosis.

Involvement of the PI3K/AKT pathway in the hypoglycemic effects of saponins from Helicteres isora.

AIM OF THE STUDY: Saponins from Helicteres isora have previously been shown to exert antidiabetic effects. The present study explored the underlying mechanisms in C2C12 skeletal muscle cells. MATERIALS AND METHODS: C2C12 cells were incubated with saponins and sapogenin followed by Western blotting and immunofluorescence analysis. RESULTS: Western blotting revealed that incubation with saponins (100 microg/ml) and sapogenin (100 microg/ml) induced the phosphorylation of the phosphatidylinositol-3-kinase (PI3K) as well as of the downstream targets protein kinase B/Akt (at Ser473) and glycogen synthase kinase GSK-3 alpha/beta (at Ser21/9) in a time-dependent manner. In contrast, no phosphorylation of the AMP-sensitive kinase AMPK (at Thr172) was observed. Within 48 h saponins/sapogenin treatment further increased the protein abundance of the insulin-sensitive glucose transporter Glut4. Confocal microscopy confirmed that saponins/sapogenin treatment stimulated Akt phosphorylation and revealed that the treatment was followed by translocation of Glut4 into the cell membrane of C2C12 muscle cells. CONCLUSIONS: Saponins and sapogenin activate the PI3K/Akt pathway thus leading to phosphorylation and inactivation of GSK-3 alpha/beta with subsequent stimulation of glycogen synthesis as well as increase of Glut4-dependent glucose transport across the cell membrane.

Liquid chromatography-mass spectrometry-based quantification of steroidal glycoalkaloids from Solanum xanthocarpum and effect of different extraction methods on their content.

A new liquid chromatography-mass spectrometry (LC-MS)-based method coupled with pressurized liquid extraction (PLE) as an efficient sample preparation technique has been developed for the quantification and fingerprint analysis of Solanum xanthocarpum. Optimum separations of the samples were achieved on a Waters MSC-18 XTerra column, using 0.5% (v/v) formic acid in water (A) and acetonitrile (ACN):2-propanol:formic acid (94.5:5:0.5, v/v/v) (B) as mobile phase. The separation was carried out using linear gradient elution with a flow rate of 1.0mL/min. The gradient was: 0min, 20% B; 14min, 30% B; 20min, 30% B; 27min, 60% B and the column was re-equilibrated to the initial condition (20% B) for 10min prior to next injection. The steroidal glycoalkaloids (SGAs) which are the major active constituents were isolated as pure compounds from the crude methanolic extract of S. xanthocarpum by preparative LC-MS and after characterization were used as external standards for the development and validation of the method. Extracts prepared by conventional Soxhlet extraction, PLE and ultrasonication were used for analysis. The method was validated for repeatability, precision (intra- and inter-day variation), accuracy (recovery) and sensitivity (limit of detection and limit of quantitation). The purpose of the work was to develop a validated method, which can be used for the quantification of SGAs in commercialized S. xanthocarpum products and the fingerprint analysis for their routine quality control.

Chromatographic analysis of Kutajarista--an ayurvedic polyherbal formulation.

Kutajarista is a well known polyherbal preparation of which the main ingredient is the stem bark of Holarrhena antidysenterica. This Ayurvedic medicine is prescribed to treat amoebic dysentery and other disorders such as fever, indigestion and malabsorption syndrome. Herbal medicines are very important since, in common with conventional medicines, they contain biologically active substances that may produce non-trivial side effects when taken in excessive amounts. Very low doses, on the other hand, may have no therapeutic value. In this paper we report the chemical standardisation of Kutajarista by HPLC analysis based upon the presence of the biomarker conessine in the formulation. The standardisation method is simple and reliable, and the precision of method has been tested for repeatability (n = 3) and reproducibility (n = 9). The response of a refractive index detector was linear in the concentration range of 0.1-1.0 mg/mL. Recovery studies were performed to check the method for accuracy. The recovery was found to be in range of 99-105%. The developed HPLC method can be used to quantify conessine for quality control of marketed Kutajarista samples.

Evaluation of proinflammatory cytokine pathway inhibitors for p38 MAPK inhibitory potential.

The target for the anti-inflammatory natural products like amentoflavone ( 2), which act by interfering with the proinflammatory cytokine pathway (e.g., TNF-alpha, IL-1beta, and NO synthase), is not yet well-defined. Data obtained from docking, electronic, and surface analyses shed some light on steric and electronic complementarity of these molecules to p38 MAPK, thereby suggesting a possible mechanism by which they might reduce the production of proinflammatory cytokines.

Postnatal development and reproductive performance of F1 progeny exposed in utero to an ayurvedic contraceptive: Pippaliyadi yoga.

Pippaliyadi yoga or pippaliyadi vati is an ayurvedic contraceptive used in India since ancient times. It is a combination of powdered fruit berries of Embelia ribes Burm.f. (Myrsinaceae), Piper longum L. (Piperaceae) and borax in equal proportion. Though the contraceptive potential is known since ancient times, no systematic developmental toxicity studies have been carried out. The present study was carried out to evaluate the postnatal developmental toxicity and the reproductive performance of the progeny exposed in utero to pippaliyadi. Pippaliyadi yoga was obtained from National Institute for Pharmaceutical Education and Research (NIPER), India and the developmental toxicity was studied by administering three doses, viz. 140, 300 and 700 mg/(kg day) to gravid females from day 6 to day 16 of gestation. Pippaliyadi did not have any adverse developmental effects with low doses, however, with the five times higher dose, a decrease in body weight of the pups was observed. The reproductive performance of the progeny born to mothers treated with pippaliyadi was not significantly affected. The present study suggests that in utero exposure to pippaliyadi does not have any adverse effect on the postnatal development and reproductive performance of the F(1) progeny.

LC and LC-MS study of stress decomposition behaviour of isoniazid and establishment of validated stability-indicating assay method.

Isoniazid was subjected to different ICH prescribed stress conditions of thermal stress, hydrolysis, oxidation and photolysis. The drug was stable to dry heat (50 and 60 degrees C). It showed extensive decomposition under hydrolytic conditions, while it was only moderately sensitive to oxidation stress. The solid drug turned intense yellow on exposure to light under accelerated conditions of temperature (40 degrees C) and humidity (75% RH). In total, three major degradation products were detected by LC. For establishment of stability-indicating assay, the reaction solutions in which different degradation products were formed were mixed, and the separation was optimized by varying the LC conditions. An acceptable separation was achieved using a C-18 column and a mobile phase comprising of water:acetonitrile (96:4, v/v), with flow rate and detection wavelength being 0.5 ml min(-1) and 254 nm, respectively. The degradation products appeared at relative retention times (RR(T)) of 0.71, 1.34 and 4.22. The validation studies established a linear response of the drug at concentrations between 50 and 1000 microg ml(-1). The mean values (+/-R.S.D.) of slope, intercept and correlation coefficient were 35,199 (+/-0.88), 114,310 (+/-4.70) and 0.9998 (+/-0.01), respectively. The mean R.S.D. values for intra- and inter-day precision were 0.24 and 0.90, respectively. The recovery of the drug ranged between 99.42 and 100.58%, when it was spiked to a mixture of solutions in which sufficient degradation was observed. The specificity was established through peak purity testing using a photodiode array detector. The method worked well on application to marketed formulation of isoniazid, and a fixed-dose combination containing isoniazid and ethambutol HCl. It was even extendable to LC-MS studies, which were carried out to identify the three degradation products. The m/z values of the peaks at RR(T) 0.71 and RR(T) 1.34 matched with isonicotinic acid and isonicotinamide, respectively. The product appearing at RR(T) 4.22 was isolated using preparative LC-MS, and turned out to be a yellow compound that was identified as isonicotinic acid N'-(pyridyl-4-carbonyl)-hydrazide based on mass, FTIR and (1)H/(13)C NMR spectral data. The same was indicated to be responsible for discolouration of isoniazid bulk drug substance and formulations, which is a familiar problem. The mechanism of formation of the said compound is outlined.

Effect of oleanane triterpenoids from Terminalia arjuna--a cardioprotective drug on the process of respiratory oxyburst.

The oleanane triterpenes arjunic acid, arjungenin and their glucosides, arjunetin and arjunglucoside II, were isolated from the bark of Terminalia arjuna. Arjungenin and its glucoside exhibited a moderate free radical scavenging activity while all the compounds showed no effect on the superoxide release from PMN cells. Further arjungenin also exhibited greater inhibitory action on the hypochlorous acid production from human neutrophils.

Ayurveda and gynecological disorders.

The science of life--Ayurveda is practiced in India since time immemorial. Besides being cheap and easily available Ayurvedic drugs are considered safe. Moreover, there is surge in the interest in Ayurveda due to quest of alternative medicines. Many of the gynecological disorders being not reported to the physicians, are treated with household remedies in India. The science of Ayurveda deals with these issues in a systematic manner as evident from the classification of diseases available and the number of plant drugs or the combinations thereof available for the treatment. In the present article, Ayurvedic herbal formulations and single plant drugs used traditionally in treatment of gynecological disorders are described.

Cyclooxygenase inhibitory flavonoids from the stem bark of Semecarpus anacardium Linn.

The ethyl acetate extract of stem bark of Semecarpus anacardium showing in vivo anti-in fl ammatory activity in carrageenan induced rat paw edema assay was investigated in order to identify its active compounds. Chemical investigation of the ethyl acetate extract of S.anacardium afforded 3,4,2',4'-tetrahydroxychalcone (butein) and 7,3',4'-trihydroxy fl avone. Evaluation of COX-1 inhibitory activity of 3,4,2',4'-tetrahydroxychalcone and 7,3',4'-trihydroxy fl avone provided the IC(50) values of 28.4 and 36.7 micro M respectively. Further investigation of these compounds for COX-2 inhibitory activity revealed moderate potency towards this enzyme.

Madhucosides A and B, protobassic acid glycosides from Madhuca indica with inhibitory activity on free radical release from phagocytes.

The structures of madhucosides A (1) and B (2), isolated from the bark of Madhuca indica, were established as 3-O-beta-D-apiofuranosyl(1-->2)-beta-D-glucopyranosyl-28-O-[beta-D-xylopyranosyl(1-->2)-[alpha-L-rhamnopyranosyl(1-->4)]-beta-D-glucopyranosyl(1--> 3)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranosyl]protobassic acid and 3-O-beta-D-apiofuranosyl(1-->2)-beta-D-glucopyranosyl-28-O-[beta-D-xylopyranosyl(1-->2)-[alpha-L-rhamnopyranosyl(1-->4)]-beta-D-glucopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranosyl]protobassic acid, respectively. These two compounds showed significant inhibitory effects on both superoxide release from polymorphonuclear cells in a NBT reduction assay and hypochlorous acid generation from neutrophils assessed in a luminol-enhanced chemiluminescence assay.

Luteinizing hormone estimation.

Luteinizing hormone (LH) is an important hormone of the reproductive system, which has found application in diagnosis and therapeutic medicine. It plays a vital role in the development and functioning of the reproductive system. Determination of LH concentration is important for detection of dysfunction of the pituitary-ovarian axis, diagnosis of reproductive disorders, monitoring of antifertility programmes and in therapeutic preparations. On the basis of heterogeneity and various biological effects served by the hormone, different assay systems have been developed for its estimation. Initially, LH was quantified on the basis of in vivo and in vitro endocrine activity. However, with the advancement in biotechnology, various immunoassays have been designed for performing most of the physiological and clinical studies on serum LH. The immunoassays offer improvement in sensitivity, precision and convenience over bioassays. However, these immunoassays have their own limitations and results obtained in different laboratories are often not comparable. This review makes an attempt to enumerate and compare various assay methods used in the serum LH measurement in varied clinical conditions.

Colonic drug delivery of 5-fluorouracil: an in vitro evaluation.

Compression coating has been found to be useful for colonic drug delivery. The aim of the present investigation was to design a formulation with a considerably reduced coat weight and gum concentration for colonic delivery of 5-fluorouracil for the treatment of colorectal cancer. Rapidly disintegrating core tablets containing 50 mg of 5-fluorouracil were prepared and compression coating with 175 mg of granules containing a mixture of xanthan gum (XG) and guar gum (GG) in varying proportions was done. With this coat weight, a highly retarded drug release was observed. After 24h of dissolution the mean percent drug release from the compression coated XG:GG 20:20, 20:10 and 10:20 tablets were found to be around 18+/-1.23%, 20+/-1.54% and 30+/-1.77%, respectively. So, the coat weight was further reduced to 150 mg. It was observed that reduction of coat weight did not affect the initial drug release rate in simulated upper gastrointestinal tract (GIT) conditions. At the end of 24h of dissolution the amount of drug released increased to 25+/-1.22%, 36.6+/-1.89% and 42.6+/-2.22%, respectively in XG:GG 20:20, 20:10 and 10:20 tablets. Studies of XG:GG (10:20) tablets in presence of colonic contents showed an increased cumulative percent drug release of 67.2+/-5.23% in presence of 2% cecal content and 80.34+/-3.89% in presence of 4% cecal content after 19 h of incubation.

A new furanoditerpenoid marker for the distinction between the seeds of two species of Caesalpinia.

A new cassane furanoditerpene, 17-methylvouacapane-8(14),-9(11)-diene (1), has been isolated from the seed kernels of Caesalpinia crista and its structure determined by mass, IR and NMR spectrometry. The 1H- and 13C-NMR spectra were assigned using a combination of 1H-1H COSY, HSQC and HMBC two-dimensional NMR experiments. An HPTLC method has been developed to quantify 1 in seed material. The furanoditerpene can serve as a marker chemically to differentiate C. crista from the synonymous C. bonduc.

Antioxidant properties of Indian medicinal plants.

The antioxidant properties of methanol extracts of 12 Indian medicinal plants, traditionally used in disease areas that probably involve free radical mechanisms, were evaluated by two methods, namely the DPPH (1,1-diphenyl-2-picryl hydrazyl) test and the lipid peroxidation assay. In the latter assay, seven of these extracts showed 90% or more activity compared with the standard, vitamin E and hence were studied in detail after the removal of interfering pigments. The selective pigment removal from the extracts led to an increase in free radical scavenging activity and a decrease in inhibition of lipid peroxidation.

Dammarane triterpene saponin from Bacopa monniera as the superoxide inhibitor in polymorphonuclear cells.

The hydroalcoholic extract of the whole plant of Bacopa monniera Wettst. (Scrophulariaceae), exhibited an inhibitory effect on superoxide released from polymorphonuclear (PMN) cells in the nitroblue tetrazolium (NBT) assay. The major saponin bacoside A(3) was found to be responsible for this effect in the herb. This compound showed 85, 91.66, 91.66, and 83 % inhibitions of NBT reduction at the concentrations of 200, 100, 50, and 25 microg/ml, respectively, with an IC(50) value of 10.22 microg/ml. These inhibitory effects were compared with those of the standard positive controls, quercetin and ascorbic acid with IC(50) of 111 and 14.16 microg/ml, respectively. Another major saponin bacopasaponin C was found to be much less potent as compared to bacoside A(3) whereas the remaining two mixtures of saponins were found to be inactive.