A blackberry-dill extract combination synergistically increases skin elasticity.

BACKGROUND: The loss of structural elastin due to intrinsic and extrinsic ageing results in the skin's inability to stretch and recoil (decrease in elasticity) and manifests as loss of skin firmness and sagging. While other extracellular matrix (ECM) components such as collagen and hyaluronic acid are continually synthesized and assembled through life, elastic fibres are not. Elastic fibre assembly and functionality require fibre cross-linking, induced by the lysyl oxidase-like (LOXL) enzymes, which sharply decrease during ageing. OBJECTIVE: To evaluate the enhanced elastogenic effect of a blackberry-dill extract combination, which was hypothesized to induce elastin fibre component synthesis, fibre cross-linking and reduce elastin fibre degradation. METHODS: The blackberry and the dill extracts were tested separately and in combination to confirm single ingredient bioactivity and synergistic benefits. Human skin explants, dermal fibroblasts, elastase assays, ELISAs, quantitative real-time PCRs and spectrofluorometer measurements were used. Moreover, a double-blinded, placebo-controlled clinical study was carried out to assess skin elasticity using Cutometer and histologically from biopsies. RESULTS: The blackberry extract induced elastin gene expression, elastin promoter activity and inhibited elastic fibre degradation by matrix metalloproteinases (MMPs) 9 and 12. The dill extract induced elastin, collagen and LOXL1 gene expression, resulting in enhanced fibre cross-linking in human skin explants. Clinically, the blackberry and dill combination treatment displayed synergistic pro-elasticity activity as compared to each ingredient alone and placebo. CONCLUSION: Taken together, these results demonstrated the two multimodal plant-based extracts complemented each other in terms of bioactivity and resulted in a synergistic elastogenesis induction.

CONTEXTE: la perte de l'élastine structurelle causée par un vieillissement intrinsèque et extrinsèque provoque l'incapacité de la peau à s'étirer et à rebondir (diminution de l'élasticité) et se manifeste comme une perte de fermeté et un relâchement de la peau. Alors que d'autres composants de la matrice extracellulaire (MEC), tels que le collagène et l'acide hyaluronique sont continuellement synthétisés et assemblés tout au long de la vie, les fibres élastiques ne le sont pas. L'assemblage et la fonctionnalité des fibres élastiques nécessitent une réticulation des fibres, causée par les enzymes de type lysyle oxydase (LOXL), qui diminuent fortement au cours du vieillissement. OBJECTIF: évaluer l'effet élastogène amélioré d'une combinaison d'extrait de mûre et d'aneth, qui était supposée induire la synthèse des composants des fibres d'élastine, la réticulation des fibres et réduire la dégradation des fibres d'élastine. MÉTHODES: les extraits de mûre et d'aneth ont été testés séparément et ensemble pour confirmer la bioactivité d'un seul ingrédient et les avantages synergiques. Des explants de peau humaine, des fibroblastes cutanés, des dosages d'élastase, des ELISA, des analyses PCR quantitatives en temps réel et des mesures de spectrofluorimètre ont été utilisés. De plus, une étude clinique en double aveugle, contrôlée par placebo, a été réalisée pour évaluer l'élasticité de la peau à l'aide du cutomètre et histologiquement à partir de biopsies. RÉSULTATS: l'extrait de mûre a induit l'expression génique de l'élastine, l'activité de promoteur de l'élastine et a inhibé la dégradation des fibres élastiques par des métalloprotéinases matricielles (MPM) 9 et 12.L'extrait d'aneth a causé l'expression génique de l'élastine, du collagène et du gène LOXL1, entraînant une amélioration de la réticulation des fibres dans les explants de peau humaine. Cliniquement, le traitement par une combinaison de mûre et d'aneth a montré une activité de pro-élasticité synergique par rapport à chaque ingrédient seul et au placebo. CONCLUSION: ensemble, ces résultats ont démontré que les deux extraits de plantes multimodales se complètent en termes de bioactivité et ont entraîné une induction synergique de l'élastogenèse.

Whole-grain intake and its inverse relationships with fatty acids intake among multiethnic Malaysian schoolchildren.

INTRODUCTION: Little is known about the relationship of whole-grain intake with dietary fatty acids intake. The present study aimed to assess the whole-grain intake and its relationships with dietary fatty acids intake among multiethnic schoolchildren in Kuala Lumpur, Malaysia. METHODS: This cross-sectional study was conducted among 392 schoolchildren aged 9-11 years, cluster sampled from five randomly selected schools in Kuala Lumpur. Whole-grain and fatty acids intakes were assessed by 3-day, 24-h diet recalls. All whole-grain foods were considered irrespective of the amount of whole grain they contained. RESULTS: In total, 55.6% (n = 218) were whole-grain consumers. Mean (SD) daily intake of whole grain in the total sample was 5.13 (9.75) g day-1 . In the whole-grain consumer's only sample, mean (SD) intakes reached 9.23 (11.55) g day-1 . Significant inverse associations were found between whole-grain intake and saturated fatty acid (SAFA) intake (r = -0.357; P < 0.001), monosaturated fatty acid (MUFA) (r = -0.373; P < 0.001) and polyunsaturated fatty acid (PUFA) (r = -0.307; P < 0.001) intake. Furthermore, whole-grain intake was a significant predictor of SAFA (ß = -0.077; P = 0.004), MUFA (ß = -0.112; P = <0.001) and PUFA (ß = -0.202; P = <0.001) intakes, after controlling for sex, age and ethnicity. CONCLUSIONS: Whole-grain intake in Malaysia was well below recommendations. Schoolchildren who consumed higher whole grain tend to reduce fat intake; however, it would also reduce the SAFA, MUFA and PUFA intakes. Future collaboration may be conducted between industry, government and universities to promote unsaturated fatty acids-rich foods and whole-grain food, although not to promote processed whole-grain foods with a high sugar and salt content.

Comparison of in vitro and in vivo fertilizing potential of buffalo bull semen frozen in egg yolk-, soya bean lecithin- and liposome-based extenders.

The objective of this study was to compare different extenders for post-thaw in vitro sperm function and in vivo fertility of buffalo semen. Accordingly, sperm of 30 ejaculates extended in egg yolk (TRIS with 20% egg yolk; EY), two soya lecithin-based (SL-1; AndroMed® and SL-2; Bioxcell® ) and a liposome-based extender (LS; OptiXcell® ) were tested. The post-thaw semen was evaluated for computer-assisted sperm analysis (CASA), sperm viability, membrane and acrosome integrity, DNA integrity and acrosome reaction and first service pregnancy rate (FSPR) in a fixed-time artificial insemination programme. Total motility and VCL were the only CASA-based parameters that exhibited significantly higher (p < .05) percentage in LS among these extenders. Post-thaw percentage of acrosome integrity (55.9 ± 1.4, 58.1 ± 2.0, 55.8 ± 2.0, 56.6 ± 2.3) and DNA integrity (68.8 ± 2.0, 69.2 ± 2.3, 71.3 ± 2.1, 69.1 ± 2.1) did not differ (p > .05) in EY, SL-1, SL-2 and LS extender, respectively. However, a variable response in terms of efficacy of different extenders for sperm viability and plasma membrane integrity was observed. Assessment of inducibility of acrosome reaction showed significant differences between extenders (51.9 ± 2.1, 44.3 ± 2.4, 46.1 ± 2.3 and 58.1 ± 3.1%, respectively, for EY, SL-1, SL-2 and LS). Furthermore, field trials revealed significantly higher (p < .05) FSPR of LS-extended semen as compared to that for EY, SL-1 and SL-2 extender (46.3%, 41.2%, 31.2% and 29.7%, respectively). It is concluded that the liposome-based extender is more effective than egg yolk- and soya lecithin-based extenders and may be used for cryopreservation of buffalo semen in the future.

Bisphenol A induced oxidative stress and apoptosis in mice testes: Modulation by selenium.

Spermatogenesis, a highly coordinated process, is prone to environmental insults which may lead to impaired spermatogenesis or, at worst, infertility. Bisphenol A (BPA) is a well-known global environmental toxicant and a ubiquitous oestrogenic chemical. This study evaluated the role of selenium (0.5 ppm sodium selenite/kg diet) on spermatogenesis after BPA treatment in different groups of male BALB/c mice: control, selenium, BPA and selenium+BPA. Markers of oxidative stress and apoptosis were evaluated in testis after BPA treatment. Significant decrease in sperm concentration and motility and increased reactive oxygen species(ROS) and LPO levels were seen in BPA group. Histopathological changes revealed extensive vacuolisation, lumen devoid of spermatozoa and decreased germ cell count, confirmed by testicular germ cell count studies. TUNEL assay for apoptosis showed increased number of TUNEL-positive germ cells in BPA group with increased percentage apoptotic index. However, in Se+BPA group, histopathological studies revealed systematic array of all germ cells, preserved basement membrane with relatively less vacuolisation, improved sperm parameters and ROS and LPO levels and decreased number of TUNEL-positive germ cells. These results clearly demonstrate the role of selenium in ameliorating oxidative stress and apoptosis induced upon BPA treatment in mice and can be further used as therapeutic target in male infertility.

Protective role of dietary-supplemented selenium and vitamin E in heat-induced apoptosis and oxidative stress in mice testes.

This study evaluated the role of selenium (0.5 ppm selenium/kg diet) and vitamin E (200 mg alpha-tocopherol/kg diet) on spermatogenesis after scrotal hyperthermia (42 °C, 30 min) in six different groups of male Balb/c mice; Control, Heat shock, Selenium, Selenium+heat shock, Vitamin E and Vitamin E+heat shock. Markers of the stress responses, hypoxia and oxidative stress, were evaluated in testis after the hyperthermic shock. Hyperthermia caused an elevated mRNA expression of hypoxia-inducible factor-1 alpha, haem oxygenase-1 (HMOX-1) and also glutathione peroxidase activity and reactive oxygen species (ROS). Apoptosis was evaluated by TUNEL assay and further by mRNA expression of Bcl-2, caspase 3, 8, 9, bid and AKT. TUNEL assay showed significant increase in apoptotic index of spermatogenic cells, whereas decrease in mRNA expression of Bcl-2, AKT and increase in caspase 3, 8, 9 and Bid in heat-shock group were observed. A significant decrease in sperm motility was also seen in heat-shock group in comparison with control group. These observations clearly indicate the development of oxidative stress and apoptosis after hyperthermia. Further analysis in Selenium+heat shock and Vitamin E+heat shock groups showed protective behaviour as compared to effects in heat-shock group which could be of therapeutic interest in future studies.

Antimalarial potential of Nosode 30 and 200 against Plasmodium berghei infection in BALB/c mice.

BACKGROUND & OBJECTIVES: Homeopathy is considered as an emerging area of alternative medicine which could be established for the global health care. One of the greatest objections to this science lies in its inability to explain the mechanism of action of the micro doses based on scientific experiments and proofs. The present study has been undertaken to screen in vivo antimalarial activity of Malaria Co Nosode 30 and Nosode 200 against Plasmodium berghei infection in BALB/c mice. METHODS: Peter's 4-day test was used to evaluate the in vivo schizontocidal effect of Nosode 30 and Nosode 200. One month follow-up study was done to calculate the mean survival time of mice in each group. Biochemical analysis was carried out to assess the liver and kidney function tests using diagnostic kits. RESULTS: Nosode 30 and 200 exhibited 87.02 and 37.97% chemosuppression on Day 7 and mean survival time (MST) of 18.5 ± 2.16 and 16.5 ± 1.37 days respectively, which were extremely statistically significant when compared to MST of infected control (8.55 ± 0.83 days). The safety of Nosode 30 was also confirmed by the comparable levels of ALP, SGOT, SGPT activities, concentration of bilirubin, urea and creatinine to CQ treated group. CONCLUSION: Nosode 30 possesses considerable in vivo antiplasmodial activity against P. berghei infection as compared to Nosode 200 as evident from the chemosuppression obtained using Peter's 4-day test. Further, studies on the drug can be carried out to establish its antimalarial potential in monotherapy or in combination with other homeopathic drug formulations.

Predischarge non-invasive risk assessment for prediction of significant hyperbilirubinemia in term and late preterm neonates.

OBJECTIVE: To evaluate efficacy of predischarge transcutaneous bilirubin (TcB) measurement and clinical risk assessment in predicting hyperbilirubinemia needing treatment. STUDY DESIGN: A diagnostic test was performed in a prospective cohort study conducted at a teaching hospital in North India. Subjects included healthy neonates with a gestation period of ≥35 weeks or birth weight ≥2000 g. Maternal, neonatal and delivery risk factors for hyperbilirubinemia were prospectively collected. TcB was measured in all enrolled neonates at 24±6, 72 to 96 and 96 to 144 h of postnatal age and when indicated clinically. Neonates were followed up during hospital stay and after discharge till completion of the 7th postnatal day. The key outcome was significant hyperbilirubinemia defined as need of phototherapy on the basis of modified American Academy of Pediatrics guidelines. In neonates born at ≥38 weeks of gestation and in neonates born at ≤37 completed weeks of gestation, middle line and lower line of phototherapy thresholds were used to initiate phototherapy, respectively. Variables observed to be significantly associated with significant hyperbilirubinemia on multivariate analysis were used for construction of a clinical risk assessment tool. Predictive ability of the risk assessment tool was assessed by calculating sensitivity, specificity, positive predictive value and negative predictive value, by plotting receiver-operating characteristics curve and calculating c-statistic. RESULT: A total of 997 neonates (birth weight: 2627±536 g, gestation: 37.8±1.5 weeks) were enrolled in the study, of which 931 completed follow-up. Among enrolled neonates, 344 (34.5%) were low birth weight. Overall, a total of 199 (20%) neonates developed significant hyperbilirubinemia. On stepwise logistic regression analysis, predischarge TcB percentile and gestation were significantly found to be associated with significant hyperbilirubinemia. A risk assessment graph was constructed to predict subsequent development of significant hyperbilirubinemia. Area under curve for this risk assessment strategy was 0.75. CONCLUSION: A risk assessment graphical tool consisting of TcB and gestation accurately predicted subsequent need of phototherapy. Further studies are needed to validate performance of this risk assessment tool.

Comparative study on the effect of chemicals on Alternaria blight in Indian mustard--a multi-location study in India.

High severity of Altemaria blight disease is a major constraint in production of rapeseed-mustard in India. The aim of this study was to investigate the suppressive potential of chemicals viz., zinc sulphate, borax, sulphur, potash and calcium sulphate, aqueous extracts viz., Eucalyptus globosus (50 g l-1) leaf extract and garlic (Allium sativum) bulb (20 g l-1) extract, cow urine and bio-agents Trichoderma harzianum, Pseudomonas fluorescence in comparison with the recommended chemical fungicide (mancozeb), against foliar disease Alternaria blight of Indian mustard [Brassica juncea (L.) Czern. and Coss] under five different geographical locations of India. Mancozeb recorded the lowest mean severity (leaf: 33.1%; pod: 26.3%) of Alternaria blight with efficacy of garlic bulb extract alone (leaf = 34.4%; pod = 27.3%) or in combination with cow urine (leaf = 34.2%; pod = 28.6%) being statistically at par with the recommended chemical fungicide. Chemicals also proved effective in reducing Alternaria blight severity on leaves and pods of Indian mustard (leaf = 36.3-37.9%; pod = 27.5-30.1%). The effective treatments besides providing significant reduction in disease severity also enabled increase in dry seed yield of the crop (mancozeb = 2052 kg ha-1; garlic = 2006 kg ha-1; control = 1561 kg ha-1).

The development of hypocretin (orexin) deficiency in hypocretin/ataxin-3 transgenic rats.

Narcolepsy is linked to a widespread loss of neurons containing the neuropeptide hypocretin (HCRT), also named orexin. A transgenic (TG) rat model has been developed to mimic the neuronal loss found in narcoleptic humans. In these rats, HCRT neurons gradually die as a result of the expression of a poly-glutamine repeat under the control of the HCRT promoter. To better characterize the changes in HCRT-1 levels in response to the gradual HCRT neuronal loss cerebrospinal fluid (CSF) HCRT-1 levels were measured in various age groups (2-82 weeks) of wild-type (WT) and TG Sprague-Dawley rats. TG rats showed a sharp decline in CSF HCRT-1 level at week 4 with levels remaining consistently low (26%+/-9%, mean+/-S.D.) thereafter compared with WT rats. In TG rats, HCRT-1 levels were dramatically lower in target regions such as the cortex and brainstem (100-fold), indicating decreased HCRT-1 levels at terminals. In TG rats, CSF HCRT-1 levels significantly increased in response to 6 h of prolonged waking, indicating that the remaining HCRT neurons can be stimulated to release more neuropeptide. Rapid eye movement (REM) sleep in TG rats (n=5) was consistent with a HCRT deficiency. In TG rats HCRT immunoreactive (HCRT-ir) neurons were present in the lateral hypothalamus (LH), even in old rats (24 months) but some HCRT-ir somata were in various stages of disintegration. The low output of these neurons is consistent with a widespread dysfunction of these neurons, and establishes this model as a tool to investigate the consequences of partial hypocretin deficiency.

Spectral integration in primary auditory cortex: laminar processing of afferent input, in vivo and in vitro.

Auditory cortex neurons integrate information over a broad range of sound frequencies, yet it is not known how such integration is accomplished at the cellular or systems levels. Whereas information about frequencies near a neuron's characteristic frequency is likely to be relayed to the neuron by lemniscal thalamocortical inputs from the ventral division of the medial geniculate nucleus, we recently proposed that information about frequencies spectrally distant from characteristic frequency is mainly relayed to the neuron via "horizontal" intracortical projections from neurons with spectrally-distant characteristic frequencies [J Neurophysiol 91 (2004) 2551]. Here we test this hypothesis by using current source density analysis to determine if characteristic frequency and spectrally-distant non-characteristic frequency stimuli preferentially activate thalamocortical and horizontal pathways, respectively, in rat auditory cortex. Characteristic frequency stimuli produced current source density profiles with prominent initial current sinks in layers 3 and 4--the termination zone of lemniscal inputs from medial geniculate nucleus. In contrast, stimuli three octaves below characteristic frequency produced initial current sinks mainly in the infragranular layers. Differences between current source density profiles were only apparent for initial current sinks; profiles for longer-latency current sinks evoked by characteristic frequency and non-characteristic frequency stimuli overlapped to a greater degree, likely due to shared mechanisms of intracortical processing or to longer-latency thalamocortical contributions (lemniscal and nonlemniscal). To identify current source density profiles produced by activation of lemniscal thalamocortical inputs alone, we utilized the mouse auditory thalamocortical slice preparation. Electrical stimulation of the medial geniculate nucleus in vitro produced major current sinks in cortical layers 3/4, and excitation spread horizontally from this point throughout primary auditory cortex to produce current sinks in multiple cortical layers. These data support the hypothesis that relay of thalamocortical information throughout auditory cortex via horizontal intracortical projections may be the basis of broad spectral integration in vivo.

Effect of enzyme supplementation on the metabolisable energy content of solvent-extracted rapeseed and sunflower seed meals for chicken, guinea fowl and quail.

(1) The nitrogen-corrected apparent metabolisable energy (AME(N)) content of solvent-extracted rapeseed and sunflower seed (un-decorticated) meals in relation to species (chicken, guinea fowl and quail) and dietary addition of feed enzymes (0 or 0.5 g/kg diet) was evaluated by a diet replacement method in a 3 x 2 factorial design. (2) The metabolism trial was conducted at two substitution levels (200 and 400 g/kg diet) of each meal with or without supplementation of commercial enzyme preparation in 6 individuals or 6 groups of cockerels, guinea fowls and quails. (3) The experimental diets were fed for a period of 12 d followed by a 3-d collection period during which total feed consumed and droppings output were quantitatively recorded. (4) The AME(N) values of rapeseed meal for cockerels, guinea fowls and quails were 8.4, 8.7 and 8.8 MJ/kg, respectively, while the corresponding values for sunflower seed meal were 6.1, 6.1 and 6.2 MJ/kg dry matter, without enzyme supplementation. (5) The AME(N) value of rapeseed meal did not improve with enzyme supplementation. However, AME(N) values of sunflower seed meal significantly increased with enzyme supplementation, from 6.1 to 6.5 MJ/kg dry matter. (6) Since AME(N) values of rapeseed meal and sunflower seed meal were similar in chicken, guinea fowl and quail, values reported for chicken could, therefore, be used for guinea fowl and Japanese quail.

Comparison of broiler chicken performance when fed diets containing meals of Bollgard II hybrid cotton containing Cry-X gene (Cry1Ac and Cry2ab gene), parental line or commercial cotton.

(1) Total and free gossypol contents were 6.2 and 0.8, 5.4 and 0.5, and 6.1 and 0.7 g/kg in meals processed (solvent extracted) from Bollgard (BG) II, non-BG II or commercial cottonseeds, respectively. (2) Broiler chicks were given one of 7 dietary treatments (iso-nitrogenous, 220 and 195 g crude protein/ kg diet at 0 to 21 and 21 to 42 d, respectively, at a metabolisable energy concentration of 12.15 MJ/kg). The treatments were: D1 (control, soybean meal [SBM] based), D2 and D3 (commercial CSM at 100 g/kg of diet with and without additional iron), D4 and D5 (BG II CSM with and without additional iron), and D6 and D7 (non-BG II parental CSM with or without additional iron). (3) Body weight gain, feed intake, feed conversion efficiency, nutrient utilisation, certain blood constituents and carcase traits were not significantly affected by dietary treatments. (4) Weights of bursa and thymus were significantly higher in groups given diets containing BG II or non-BG diets containing added iron. (5) The results suggest that low free gossypol content cottonseed meals, for example, BG II, non-BG II and commercial solvent-extracted CSM could be included at 100 g/kg in broiler diets, safely replacing soybean meal without additional iron.

The in vitro antimutagenic activity of Triphala--an Indian herbal drug.

A study to evaluate an antimutagenic potential of water, chloroform and acetone extracts of Triphala has been made in an Ames histidine reversion assay using TA98 and TA100 tester strains of Salmonella typhimurium against the direct-acting mutagens, 4-nitro-o-phenylenediamine (NPD) and sodium azide, and the indirect-acting promutagen, 2-aminofluorene (2AF), in the presence of phenobarbitone-induced rat hepatic S9. A combination drug 'Triphala' - a composite mixture of Terminalia bellerica, T. chebula and Emblica officinalis, has been used in traditional system of medicine for the treatment of many malaises, such as heart ailments and hepatic diseases. The drug was sequentially extracted with water, acetone and chloroform at room temperature. The study revealed that water extract was ineffective in reducing the revertants induced by the mutagens. The results with chloroform and acetone extracts showed inhibition of mutagenicity induced by both direct and S9-dependent mutagens. A significant inhibition of 98.7% was observed with acetone extract against the revertants induced by S9-dependent mutagen, 2AF, in co-incubation mode of treatment. Various spectroscopic techniques, namely 1H-NMR, normal 13C-NMR, distortionless enhancement by polarization transfer (DEPT-90 and DEPT-135), UV and IR, are under way to identify the polyphenolic compounds from an acetone extract.

GC-MS analysis of essential oils of heartwood and resin of Shorea robusta.

The essential oils of Shorea robusta heartwood and resin were isolated by hydrodistillation of their respective petroleum ether extracts. Nine and seventeen compounds representing 80.35 % and 78.43 % of the oil, respectively were identified by GC-MS. Germacrene-D was found to be the chief constituent of both the oils. This is the first report on heartwood and resin oils of Shorea robusta.

Antimutagenic potential of extracts isolated from Terminalia arjuna.

Terminalia arjuna is an important medicinal plants widely used in the preparation of Ayurvedic formulations used against several ailments. The present investigation was aimed at the fractionation of crude extracts from the bark of T. arjuna in order to isolate and purify the antimutagenic factors present. The antimutagenicity assay was performed to check the modulatory effect of these fractions against NPD, sodium azide, and 2AF, using the Ames Salmonella his+ reversion assay. Most of the phenolic fractions exhibited mutagen specificity against direct-acting mutagens, being effective in suppressing the frameshift mutagen NPD but failing to inhibit sodium azide (base pair substitution)-induced his+ revertants. ET-1 fraction triterpenoid diglycoside showed a marked effect against sodium azide but was ineffective against NPD. In the case of the indirect-acting mutagen 2AF, all the fractions were found to be quite potent in modulating its mutagenicity in both TA98 and TA100 tester strains of Salmonella typhimurium. The results indicate that the bark of T. arjuna harbors constituents with promising antimutagenic/anticarcinogenic potential that should be investigated further.

Modulatory effects of a tannin fraction isolated from Terminalia arjuna on the genotoxicity of mutagens in Salmonella typhimurium.

A fraction isolated from Terminalia arjuna was studied for its antimutagenic effect against 4-nitro-o-phenylenediamine (NPD) in TA98, sodium azide in TA100 and 2-aminofluorene (2AF, S9-dependent), a promutagen, in both TA98 and TA 100 tester strains of Salmonella typhimurium using the Ames assay. The fraction inhibited the mutagenicity of 2AF very significantly in both strains while the revertant colonies induced by NPD and sodium azide were reduced moderately. 1H-NMR, 13C-NMR, IR and UV-spectroscopic data of the fraction revealed it to be tannin in nature.

Growth suppression of human transformed cells by treatment with bark extracts from a medicinal plant, Terminalia arjuna.

We have investigated the effects of acetone and methanol extracts of a medicinal plant, Terminalia arjuna, on the growth of human normal fibroblasts (WI-38), osteosarcoma (U2OS), and glioblastoma (U251) cells in vitro. We found that both extracts at 30 microg and 60 microg/ml concentrations inhibit the growth of transformed cells; the growth of normal cells was least affected. Although the transformed cells appeared to have fragmented nucleus by Hoechst staining, no deoxy-ribonucleic acid laddering effect was observed. In response to the extract treatment, the tumor suppressor protein, p53, was induced in U2OS but not in U251 and WI-38 cells. A cyclin-dependent kinase inhibitor, p21WAF1, was induced in transformed cells only. The study suggests that the bark extract of medicinal plant, T. arjuna, has components that can induce growth arrest of transformed cells by p53-dependent and -independent pathways.

Antimutagenicity of hydrolyzable tannins from Terminalia chebula in Salmonella typhimurium.

A tannin fraction (TC-E) from the dried fruit pulp of Terminalia chebula was obtained by successfully extracting with 95% ethyl alcohol and ethyl acetate. TC-E was subjected to silica gel chromatography which yielded four fractions, viz., TC-EI, TC-EII, TC-EIII and TC-EIV. Thin layer chromatography (TLC) and 13C-NMR revealed that TC-EI was gallic acid (GA) derivative while the other fractions were tannin in nature. TC-E and its fractions were evaluated for their antimutagenic potential against two direct-acting mutagens, 4-nitro-o-phenylenediamine (NPD) and 4-nitroquinoline-N-oxide (4NQNO), and S9-dependent mutagen, 2-aminofluorene (2AF) in TA98 and TA100 strains of Salmonella typhimurium. The study revealed that the extract (TC-E) and its fractions were highly significant against S9-dependent mutagen, 2AF. The effect was found to be more or less corresponding with the nature of the fractions, as the monomeric TC-EI (a GA derivative) was least effective as compared to other fractions which were oligomeric, and the order of their effectiveness as per their IbD50 value being TC-EIV (8.9 micrograms)>TC-EIII (17.8 micrograms)>TC-EII (45 micrograms)>TC-EI (320 micrograms) in TA98; TC-EIV being 40 times more effective than TC-EI in inhibiting his+ revertants. A similar effect was noticed in TA100 too, where TC-EI was the least effective and TC-EII had the maximum effect. A similar result was noticed when the antimutagenicity of GA (a monomeric) was compared with tannic acid (TA, an oligomeric). However, chebula tannins were found to be partly effective against NPD but not at all effective against 4NQNO.

MK 801 reverses haloperidol-induced catalepsy from both striatal and extrastriatal sites in the rat brain.

The present study investigated whether the anticataleptic effect of (+)-5-methyl-10,11-dihydro-5H-dibenzo(a,d)-cyclohepten-5,10-imine (MK 801) is due to a blockade of N-methyl-D-aspartate (NMDA) receptors in striatal output pathways as well as in the striatum. Catalepsy induced by haloperidol (1 mg/kg i.p.) was more effectively reversed by MK 801 (0.2 mg/kg i.p.) given 10 min prior to rather than 45 min after the neuroleptic. Catalepsy evoked by intrastriatal haloperidol (7 micrograms/side) was also strongly attenuated by systemic MK 801 (0.2 mg/kg i.p.). We also found that the cataleptic rigidity induced by systemic haloperidol (1 mg/kg i.p.) could be prevented by prior injection of MK 801 into the striatum (10 micrograms), subthalamic nucleus (5 micrograms), entopeduncular nucleus (5 micrograms) or substantia nigra pars reticulata (1 microgram). These results suggest that the anticataleptic action of systemic MK 801 versus haloperidol, is due to the blockade of NMDA receptors in the striatum as well as in striatal output circuits through the subthalamus. However, systemic MK 801 (0.2 mg/kg i.p.) was without effect on the catalepsy elicited by injecting muscimol into the globus pallidus (25 ng) or ventromedial thalamus (50 ng). These findings suggest that MK 801 has little influence over thalamic excitatory feedback to the cortex, and that hypoactivity of the pallidum may not be a prerequisite for hyperactivity in the subthalamus.

Affinity selection and mass spectrometry-based strategies to identify lead compounds in combinatorial libraries.

The screening of diverse libraries of small molecules created by combinatorial synthetic methods is a recent development which has the potential to accelerate the identification of lead compounds in drug discovery. We have developed a direct and rapid method to identify lead compounds in libraries involving affinity selection and mass spectrometry. In our strategy, the receptor or target molecule of interest is used to isolate the active components from the library physically, followed by direct structural identification of the active compounds bound to the target molecule by mass spectrometry. In a drug design strategy, structurally diverse libraries can be used for the initial identification of lead compounds. Once lead compounds have been identified, libraries containing compounds chemically similar to the lead compound can be generated and used to optimize the binding characteristics. These strategies have also been adopted for more detailed studies of protein-ligand interactions.