FAS and SREBP-1c Inhibition via AMPK Activation in HepG2 Cells by Biovip, Tox-off, and Traphanoside GO1 from Glinus oppositifolius.

Glinus oppositifolius is an endemic herbaceous plant found in tropical Asian countries and is native in Vietnam. It is used in traditional folk medicine because of its flavor and antiseptic and laxative effects. In the current research, the effects of Tox-off, Biovip, and the purified compounds isolated from G. oppositifolius in the previous study were evaluated on the activation of adenosine 5'-monophosphate-activated protein kinase (AMPK)-activated protein kinase (AMPK) and acetyl-coenzyme A carboxylase (ACC) in C2C12 myoblasts. In addition, the most potent active compounds, traphanoside-GO1 (TRA-GO1) and TRA-GO5 have validated the reduction of fatty acid synthase (FAS) and sterol regulatory element binding protein (SREBP)-1c in HepG2 cells. We found that Tox-off and Biovip significantly increased the phosphorylation of AMPK and ACC in C2C12 myoblasts. Furthermore, TRA-GO1 and TRA-GO5 significantly increased the AMPK activation and phosphorylation of its downstream substrate ACC in a concentration-dependent way compared to the dimethyl sulfoxide (DMSO) control. Besides, the protein level of FAS and SREBP-1c decreased by TRA-GO1 and TRA-GO5 in a concentration-dependent manner. Taken together, our results showed that the increased AMPK and ACC phosphorylation by active components of G. oppositifolius may activate the AMPK signaling pathways, which are useful for the anti-obesity and its related metabolic disorders.

New Triterpenoids and Anti-Inflammatory Constituents from Glinus oppositifolius.

Three new triterpenoids-spergulagenin B (1), spergulagenin C (2), and spergulagenin D (3)-were isolated from the aerial part of Glinus oppositifolius, along with 17 known compounds (4-20). The structures of these new compounds were identified by spectroscopic and MS analyses. Compounds 3, 5, 19, and 20 were evaluated for inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells with IC50 values of 17.03, 18.21, 16.30, and 12.64 µM, respectively. Compounds 3, 5, and 20 exhibited inhibitory effects on LPS-induced nitric oxide production in RAW 264.7 cells with IC50 values of 18.35 ± 1.34, 17.56 ± 1.41, and 14.27 ± 1.29 µM, respectively.

Triterpenoids and Their Glycosides from Glinus Oppositifolius with Antifungal Activities against Microsporum Gypseum and Trichophyton Rubrum.

Four new triterpenoids, 3ß,12ß,16ß,21ß,22-pentahydroxyhopane (1), 12ß,16ß,21ß,22-tetrahydroxyhopan-3-one (2), 3-oxo-olean-12-ene-28,30-dioic acid (3), and 3ß-hydroxyoleana-11,13(18)-diene-28,30-dioic acid 30-methyl ester (4); 21 new triterpenoid saponins, glinusopposides A-U (5-25); and 12 known compounds (26-37) were isolated from the whole plants of Glinus oppositifolius. The structures of the new compounds were elucidated based on the analysis of one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) and mass spectrometry (MS) data. All compounds from the plants were measured for antifungal activities against Microsporum gypseum and Trichophyton rubrum. Glinusopposide B (6), glinusopposide Q (21), glinusopposide T (24), and glinusopposide U (25) showed strong inhibitory activities against M. gypseum (MIC50 7.1, 6.7, 6.8, and 11.1 µM, respectively) and T. rubrum (MIC50 14.3, 13.4, 11.9, and 13.0 µM, respectively). For those active compounds with an oleanane skeleton, glycosylation (21-26) or oxidation (3) of 3-OH was helpful in increasing the activity; replacement of the 30-methyl group (29) by a carboxymethyl group (26) enhanced the activity; the presence of 11,13(18) double bonds (20) decreased the activity.

Anti-inflammatory and anti-osteoarthritis effect of Mollugo pentaphylla extract.

CONTEXT: Mollugo pentaphylla L. (Molluginaceae) extract (MPE) has been reported to have anti-inflammatory effect on MSU-induced gouty arthritis in a mouse model. OBJECTIVE: This study examined the anti-inflammatory activities of an MPE in vitro and anti-osteoarthritis effects on monosodium iodoacetate (MIA)-induced osteoarthritis (OA) in vivo. MATERIALS AND METHODS: The dried whole plants of M. pentaphylla were extracted with 70% ethanol under reflux. The anti-inflammatory effect of MPE was evaluated in vitro in lipopolysaccharide (LPS)-treated RAW264.7 cells. The anti-osteoarthritic effect of MPE was investigated in a Sprague-Dawley rat model of MIA-induced OA. Each seven male rats were orally administered MPE (75, 150 or 300 mg/kg) or the positive control drug indomethacin (1 mg/kg) 3 days before MIA injection and once daily for 11 days thereafter. After the treatment with MPE, no evidence of systemic adverse effects was observed in any study group. RESULTS: MPE exhibited anti-inflammatory activity via inhibition of the production of NO (57.8%), PGE2 (97.1%) and IL-6 (93.2%) in LPS-treated RAW264.7 cells at 200 µg/mL. In addition, MPE suppressed IL-1ß (60.9%), TNF-α (37.9%) and IL- 6 (40.9%) production and suppressed the synthesis of MMP-2, MMP-9 and COX-2 in the MIA-induced OA rat model. CONCLUSIONS: These results demonstrate that MPE exerts potent anti-inflammatory activities and protects cartilage in an OA rat model. This might be a potential candidate for therapeutic OA treatment.

Effects of Mollugo pentaphylla extract on monosodium urate crystal-induced gouty arthritis in mice.

BACKGROUND: Gout is an inflammatory condition induced by the deposition of monosodium urate (MSU) crystals in joints and soft tissues, and it can lead to acute or chronic arthritis. MSU are pro-inflammatory stimuli that can initiate, amplify and sustain an intense inflammatory response. In this study, we evaluated the anti-inflammatory effect of an extract of Mollugo pentaphylla (MPE) on MSU-induced gouty arthritis in a mouse model. METHOD: An MSU crystal suspension (4 mg/50 µL) was injected intradermally into the right paw. The mice were orally administered MPE (150 mg/kg or 300 mg/kg) or the positive control drug colchicine (1 mg/kg) 1 h before the MSU crystals were injected and then once daily for 3 days. The effects of MPE included inflammatory paw edema and pain upon weight-bearing activity, and we evaluated the inflammatory cytokine expression and paw tissue inflammation-related gene expression. RESULTS: MPE suppressed inflammatory paw edema and pain in the MSU-induced mice. MPE showed anti-inflammatory activity by inhibiting the production of TNF-α, interleukin (IL)-1ß, NLRP3 inflammasome and NF-κB. CONCLUSION: These results suggest that MPE has potent anti-inflammatory activities and may be useful as a therapeutic agent against gouty arthritis.

Generic relationships among Molluginaceae inferred from a molecular phylogenetic analysis of the matK gene.

The family Molluginaceae (order Caryophyllales) is considered polyphyletic based on the photosynthetic pathway, C4 evolution, and phylogeny of the family. This inference was made based on photosynthetic, anatomical, and molecular datasets. The generic circumscription of this family has greatly been changed owing to the placement of several of its genera into the Caryophyllaceae, Microteaceae, Lophiocarpaceae, and Limeaceae families. However, the generic relationships are largely unknown. By virtue of high substitution rates within the species and the ability to resolve the phylogenetic position of morphologically very closely related species and species complexes, the matK gene has emerged as one of the potential chloroplast DNA molecular markers in plant molecular phylogenetics and DNA barcoding studies. We herein used molecular phylogenetic analyses of matK gene sequences using maximum parsimony and maximum likelihood analyses to infer the generic relationships among currently recognized genera circumscribed under the family Molluginaceae. The resulting phylogenetic tree confirmed the polyphyly of the family Molluginaceae. The genus Hypertelis was found at the base of the Molluginaceae clade. The genus Glinus was close to Glischrothamnus and Mollugo, Suessenguthiella was close to Coelanthum and Pharnaceum, whereas Polpoda grouped with Adenogramma and Psammotropha. The present study constitutes a robust investigation of the molecular phylogenetic relationships among members of the family Molluginaceae. Future study should combine by combined analyses of morphological characters and multiple nuclear and chloroplast DNA sequences with a more comprehensive taxon sampling of the family Molluginaceae.

Recent progress in Glinus oppositifolius research.

CONTEXT: Glinus oppositifolius (L.) Aug. DC. (Molluginaceae), a perennial subshrubs herb, grows at low altitudes in the southern part of Taiwan, and is used in traditional Chinese medicine for herpes zoster and herpangina. OBJECTIVE: This study describes nutritional and therapeutic potential of Glinus oppositifolius and summarizes scientific evidence that supports traditional claims; recent progress in research for this plant is reviewed herein. MATERIALS AND METHODS: The literature has been retrieved from the web-based online systems including PubMed, Medline, and Google Scholar. The articles related to phytochemistry, pharmaceutical biology and ethnopharmacology have been excluded. RESULTS AND DISCUSSION: In clinical practice, the plant has been extensively investigated in a broad range of studies to provide scientific evidence for folklore claims or to find new therapeutic uses. The present review may arouse related research and make a more valid display for Taiwanese native medicinal plants.

Chemometric profiling and anti-arthritic activity of aerial parts of Glinus oppositifolius (L.) Aug. DC.

ETHNOPHARMACOLOGICAL RELEVANCE: Glinus oppositifolius (L.) Aug. DC. belongs to the family Molluginaceae, an annual prostrate herb traditionally used to treat inflammations, arthritis, malarial, wounds, fevers, diarrhoea, cancer, stomach discomfort, jaundice, and intestinal parasites. However, the anti-arthritic activity of the aerial part has still not been reported. AIM OF THE STUDY: To investigate the antioxidant and anti-arthritic activity of G. oppositifolius in Complete Freund's Adjuvant (CFA) induced rats. MATERIALS AND METHODS: The dried aerial parts of this plant material were defatted with n-hexane and extracted by methanol using a soxhlet apparatus. The in vitro anti-arthritic activity of methanolic extract of G. oppositifolius (MEGO) was evaluated in protein denaturation, membrane stabilization, and inhibition of proteinase assay at 25, 50, 100, 200, and 400 µg/ml concentrations. Female Wistar rats were immunized sub-dermally into the right hind paw with 0.1 ml of CFA. Rats were administered with MEGO at doses of 200 and 400 mg/kg once daily for fourteen days after arthritis induction. Assessment of arthritis was performed by measuring paw diameter, arthritic index, arthritic score, body weight, organ weight, and hematological and biochemical parameters, followed by the analysis of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interleukin-1-beta (IL-1ß), cyclooxygenase-2 (COX-2), interleukin 13 (IL-13) and interleukin 10 (IL-10) and histopathological study. In vivo antioxidant effect was investigated in enzymatic assays. The presence of phytoconstituents was analyzed by Gas Chromatography-Mass Spectrometry (GC-MS) and Liquid Chromatography-Mass Spectrometry (LC-MS), respectively. In silico molecular docking study of the compounds was carried out against COX-2, IL-1ß, IL-6, and TNF-α using AutoDock 4.2 and BIOVIA-Discovery Studio Visualizer software. RESULTS: MEGO's in vitro anti-arthritic activity showed dose-dependent inhibition of protein denaturation, membrane stabilization, and proteinase inhibition, followed by significant in vivo anti-arthritic activity. The rats treated with MEGO showed tremendous potential in managing arthritis-like symptoms by restoring hematological, biochemical, and histological changes in CFA-induced rats. MEGO (200 and 400 mg/kg) showed a significant alleviation in the levels of hyper expressed inflammatory mediators (TNF-α, IL-1ß, and IL-6) and oxidative stress (SOD, CAT, GSH, and LPO) in CFA-induced rats. Spergulagenin-A as identified by LC-MS analysis, exhibited the highest binding affinity against COX-2 (-8.6), IL-1ß (7.2 kcal/mol), IL-6 (-7.4 kcal/mol), and TNF-α (-6.5 kcal/mol). CONCLUSIONS: Provided with the comprehensive investigation, methanolic extract of G. oppositifolius against arthritic-like condition is a proof of concept that revalidates its ethnic claim. The presence of Spergulagenin-A might be responsible for the anti-arthritic activity.

Androecial evolution in Caryophyllales in light of a paraphyletic Molluginaceae.

PREMISE OF THE STUDY: Caryophyllales are highly diverse in the structure of the perianth and androecium and show a mode of floral development unique in eudicots, reflecting the continuous interplay of gynoecium and perianth and their influence on position, number, and identity of the androecial whorls. The floral development of five species from four genera of a paraphyletic Molluginaceae (Limeum, Hypertelis, Glinus, Corbichonia), representing three distinct evolutionary lineages, was investigated to interpret the evolution of the androecium across Caryophyllales. • METHODS: Floral buds were dissected, critical-point dried and imaged with SEM. The genera studied are good representatives of the diversity of development of stamens and staminodial petaloids in Caryophyllales. • KEY RESULTS: Sepals show evidence of petaloid differentiation via marginal hyaline expansion. Corbichonia, Glinus, and Limeum also show perianth differentiation via sterilization of outer stamen tiers. In all four genera, stamens initiate with the carpels and develop centrifugally, but subsequently variation is significant. With the exception of Limeum, the upper whorl is complete and alternisepalous, while a second antesepalous whorl arises more or less sequentially, starting opposite the inner sepals. Loss or sterilization of antesepalous stamens occurs in Glinus and Limeum and is caused by altered carpel merism and inhibition by sepal pressures. • CONCLUSIONS: Outer stamens of Hypertelis correspond with petaloids of Caryophyllaceae and suggest that staminodial petaloids and outer alternisepalous stamens are interchangeable in the Caryophyllales. We emphasize a switch in the position of first formed stamens from antesepalous to alternisepalous following the divergence of Limeum; thus stamen position is an important synapomorphy for the globular inclusion clade.

An in vitro study on the DNA damaging effects of phytochemicals partially isolated from an extract of Glinus lotoides.

An extract of Glinus lotoides, a medicinal plant used in Africa and Asia for various therapeutic purposes, was recently shown to cause DNA damage in vitro. To further explore the potential genotoxicity of this plant, fractionation of the crude extract was performed using reverse phase solid-phase extraction and a stepwise gradient elution of methanol in water. Four fractions were collected and subsequently analysed for their DNA damaging effects in mouse lymphoma cells using an alkaline version of the comet assay. To identify potential genotoxic and non-genotoxic principles, each fraction was then subjected to liquid chromatography coupled to mass spectrometry, LC-MS/MS. 1D and 2D nuclear magnetic resonance analyses were used to confirm the identity of some saponins. Although fractions containing a mixture of flavonoids and oleanane-type saponins or oleanane-type saponins alone produced no DNA damage, those containing hopane-type saponins exhibited a pronounced DNA damaging effect without affecting the viability of the cells. To conclude, even if this study presents evidence that hopane-type of saponins are endowed with a DNA damaging ability, further studies are needed before individual saponins can be cited as a culprit for the previously reported genotoxicity of the crude extract of G. lotoides.

Biosynthesis of Co3O4 nanomedicine by using Mollugo oppositifolia L. aqueous leaf extract and its antimicrobial, mosquito larvicidal activities.

Nanotechnology is a relatively revolutionary area that generates day-to-day advancement. It makes a significant impact on our daily life. For example, in parasitology, catalysis and cosmetics, nanoparticles possess distinctive possessions that make it possible for them in a broad range of areas. We utilized Mollugo oppositifolia L. aqueous leaf extract assisted chemical reduction method to synthesize Co3O4 nanoparticles. Biosynthesized Co3O4 Nps were confirmed via UV-Vis spectroscopy, scanning electron microscope, X-ray diffraction, EDX, Fourier-transform infrared, and HR-TEM analysis. The crystallite size from XRD studies revealed around 22.7 nm. The biosynthesized Co3O4 nanoparticle was further assessed for mosquito larvicidal activity against south-urban mosquito larvae Culex quinquefasciatus, and antimicrobial activities. The synthesized Co3O4 particle (2) displayed significant larvicidal activity towards mosquito larvae Culex quinquefasciatus with the LD50 value of 34.96 µg/mL than aqueous plant extract (1) and control Permethrin with the LD50 value of 82.41 and 72.44 µg/mL. When compared to the standard antibacterial treatment, Ciprofloxacin, the Co3O4 nanoparticle (2) produced demonstrates significantly enhanced antibacterial action against the pathogens E. coli and B. cereus. The MIC for Co3O4 nanoparticles 2 against C. albicans was under 1 µg/mL, which was much lower than the MIC for the control drug, clotrimale, which was 2 µg per milliliter. Co3O4 nanoparticles 2, with a MIC of 2 µg/mL, has much higher antifungal activity than clotrimale, whose MIC is 4 µg/mL, against M. audouinii.

A new triterpenoid saponin from Glinus oppositifolius.

From the ethanol extract of Glinus oppositifolius, collected at Phu Yen province, Viet Nam, one new triterpenoid saponin (1) and four known compounds (2-5) were isolated. By means of NMR and HR-ESI-MS analyses, their structure was elucidated as 3-O-(ß-D-xylopyranosyl-(1→3)-ß-D-xylopyranosyl)spergulagenin A or glinusopposide V (1), glinusopposide L (2), spergulin B (3), vitexin (4) and astralagin (5). Two compounds (1-2) showed weak inhibitory activity against α-glucosidase.

Anti-inflammatory Activity of Mollugo cerviana Methanolic Extract in LPS-induced Acute Inflammatory RAW 264.7 Macrophages.

BACKGROUND: The management of acute inflammation, which arises from complex biological responses to harmful stimuli, is an important determinant in the recovery from an otherwise detrimental outcome such as septicemia. However, the side effects and limitations of current therapeutics necessitate the development of newer and safer alternatives. Mollugo cerviana is a common medicinal herb of the Indian subcontinent and has been traditionally used for its fever mitigating anti-microbial and hepatoprotective action. We have already reported the rich presence of radical scavenging phytochemicals in the plant extracts and their strong antioxidant properties. OBJECTIVE: In the present study, we have evaluated the anti-inflammatory effects of methanolic extract (ME) of the areal parts of M. cerviana in a lipopolysaccharide (LPS)-induced acute inflammatory cell culture model. METHODS: RAW 264.7 mouse macrophage cells were stimulated by the bacterial endotoxin LPS at a concentration of 1 µg/mL. Cytotoxicity and anti-inflammatory potential of ME were carried out. RESULTS: The concentration of M. cerviana extract up to 150 µg/ml was found to be non-toxic to cells (MTT and NRU assay). LPS induces acute inflammation by binding to TLR-4 receptors, initiating a downstream signalling cascade that results in pro-inflammatory cytokine secretion. Extract treatment at 100 µg/ml suppressed LPS-induced gene expression (qPCR) and secretion (ELISA) of pro-inflammatory cytokines IL-1ß, IL-6 and TNF-α, and the chemokine CCL2, leading to dampening of the acute inflammatory cascade. LPS-induced elevation of ROS level (DCFDA method) was significantly reduced by extract treatment. Nitric oxide production, as indicated by nitrite level, was significantly reduced post extract treatment. CONCLUSION: This study demonstrated that M. cerviana methanolic extract has a potent antiinflammatory effect in the in vitro acute inflammation model of LPS-stimulated RAW 264.7 cells. There is no reported study so far on the anti-inflammatory properties of M. cerviana in an LPSinduced acute inflammatory model, which closely mimics a human bacteremia response. Hence, this study highlights the therapeutic potential of this extract as a source of anti-inflammatory lead molecules.

Traphanoside GO1, a new triterpenoid saponin from the aerial parts of Glinus oppositifolius with the inhibitory effect on PGE2 production in LPS-induced HepG2 cells.

A new saponin, 3-O-[α-ʟ-rhamnosyl-(1→3)-ß-D-glucopyranosyl]-28-O-ß-D-glucopyranosyl serjanic acid (Traphanoside GO1, 11) along with eleven compounds (1-10 and 12) were isolated from the aerial parts of Glinus oppositifolius. The structures of all isolates were elucidated by analyzing extensive 1 D- and 2 D-NMR and HR-ESI-MS, comparing with reported literature data. Compounds 7-8, 10-11, and 90% ethanol extract (GOE90) were evaluated for the inhibitory effect on PGE2 production from activated HepG2 cells. Among these, new compound 11 showed the most potent inhibitory activity by suppressing LPS-induced PGE2 production on the HepG2 cells.

L-(-)-(N-trans-cinnamoyl)-arginine, an acylamino acid from Glinus oppositifolius (L.) Aug. DC.

An amino acid derivative, L-(-)-(N-trans-cinnamoyl)-arginine, was isolated from the whole plant of Glinus oppositifolius (L.) Aug. DC. along with kaempferol 3-O-galactopyranoside, isorhamnetin 3-O-beta-D-xylopyranosyl-(1-->2)-beta-D-galactopyranoside, vitexin, vicenin-2, adenosine and L-phenylalanine. The structure determinations were based on analyses of chemical and spectroscopic methods.

Antibacterial and anti-inflammatory effects of Jeju medicinal plants against acne-inducing bacteria.

Propionibacterium acnes and Staphylococcus epidermidis are pus-forming bacteria that trigger inflammation in acne. The present study was conducted to evaluate the antimicrobial activities of Jeju medicinal plants against these etiologic agents of acne vulgaris. Ethanol extracts of Jeju plants were tested for antimicrobial activities by disc diffusion and broth dilution methods. The results from the disc diffusion assays revealed that four medicinal plants, Mollugo pentaphylla, Angelica anomala, Matteuccia orientalis, and Orixa japonica inhibited the growth of both pathogens. Among these, A. anomala had strong inhibitory effects. Its MIC values were 15.6 microg/ml and 125 microg/ml against P. acnes and S. epidermidis, respectively. The cytotoxic effects of the four extracts were determined by colorimetric MTT assays using two animal cell lines: human dermal fibroblasts and HaCaT cells. Although the M. orientalis root extract had moderate cytotoxicity in HaCaT cells at 200 microg/ml, most extracts exhibited low cytotoxicity at 200 microg/ml in both cell lines. In addition, the extracts reduced the P. acnes-induced secretion of interleukin-8 and tumor necrosis factor-alpha (TNF-alpha) in THP-1 cells, an indication of their anti-inflammatory effects. Based on these results, we suggest that M. pentaphylla, A. anomala, M. orientalis, and O. japonica are attractive acne-mitigating candidates for topical application.

Granulation by roller compaction and enteric coated tablet formulation of the extract of the seeds of Glinus lotoides loaded on Aeroperl 300 Pharma.

The purpose of this research was to improve the hygroscopicity and poor flow properties of the crude dry extract of the seeds of Glinus lotoides and improve the disintegration time of the core-tablets for enteric coated formulation thereof. The liquid crude extract of the plant was adsorbed on granulated colloidal silicon dioxide (Aeroperl 300 Pharma) at 30% w/w and the dry extract preparation (DEP) was dry-granulated with roller-compaction using Micro-Pactor. Hygroscopicity, flow property and disintegration time were improved significantly due to the adsorption and granulation processes. Moreover, the DEP does not become mucilaginous even at higher relative humidity levels (above 65%). Oblong tablets (20 x 8.25 mm) containing 947 mg of the granulated DEP (equivalent to the traditional dose), 363 mg of Avicel PH101 and 90 mg of Ac-di-Sol as disintegrant were formulated using an instrumented eccentric tablet machine at 20 kN. The tablets showed a crushing strength of 195 N, a friability of 0.4% and disintegrated within 9 min. The tablets were then enteric coated using polymethacrylate co-polymers (Eudragit L 100-55 and Kollicoat MAE 100P). The coated tablets resisted disintegration or softening in simulated gastric fluid for a minimum of 2 h and disintegrated within 15 min in intestine simulated fluid at pH 6.8. In addition to controlling the release of the active agents, the enteric coating improved the strength and decreased friability of the core-tablets.

Toxicological study on Glinus lotoides: a traditionally used taenicidal herb in Ethiopia.

Seeds of Glinus lotoides L. (Molluginaceae) are used traditionally in the treatment of tapeworm infestation in Ethiopia. Previous studies on its anthelmintic activities confirmed its traditional claims, but data on safety profile were lacking. To this effect, single and repeated dose oral toxicities of the methanolic extracts of seeds of Glinus lotoides were conducted in rats. Doses of 0, 1000 and 5000 mg/kg of crude extract of Glinus lotoides were employed in single dose toxicity studies, while doses of 0, 250, 500, and 1000 mg/kg were used in repeated toxicity studies. In the single dose toxicity test, oral administration of 5000 mg/kg of Glinus lotoides produced mortality in two females and one male on day 4. No significant differences in body and organ weights were observed between controls and treated surviving animals. Moreover, both gross and microscopic examinations of organs did not show detectable differences between controls and treated animals of both sexes. In repeated dose toxicity studies, no mortality was observed when varying doses of the extracts were administered per day for a period of 28 days. There were no significant differences in body weight, absolute and relative organ weights between controls and treated animals of both sexes. Hematological analysis showed no differences in most parameters examined. In the biochemistry parameter analysis, no significant change occurred. Pathologically, neither gross abnormalities nor histopathological changes were observed. These finding suggest that none of the organs appeared to be target and the data could provide satisfactory preclinical evidence of safety to launch clinical trial on standardized formulation of plant extracts.

Thermal Performance Curves Reveal Variation in the Seasonal Niche of a Short-Lived Annual.

An organism's environment can vary over spatial and temporal scales. Seasonal variation is an important but overlooked source of environmental variation that often shapes the ranges of organisms. The seasonal niche is a description of the spatiotemporal range of an organism resulting from spatial variation in seasonal conditions. In this study, I describe the seasonal niche of a short-lived annual plant, and variation within the species in seasonal niche breadth. I construct a seasonal species distribution model (SDM) for the species, and using thermal performance curves (TPCs), construct mechanistic SDMs (MSDMs) for individual genotypes. I quantify the correlation between the suitability scores generated in the SDM and the predicted dry weight generated by the MSDMs for each genotype, to estimate variation in seasonal niche breadth among genotypes. Thus, the parameters of TPCs reflect generalist/specialist strategies. I detected significant relationships between thermal performance breadth and maximum predicted fitness and significant correlations between optimal growth temperature and thermal performance breadth. There were large positive correlations between predictions of the SDM and MSDMs based on growth within individual genotypes. The variation in these correlations suggests variation in the degree of specialization. Genotypes with the broadest TPCs had the largest correlations between their MSDMs and the SDM, suggesting that they were generalists. The results show that correlative and MSDMs make similar predictions over the seasonal range, and that ecological specialization can vary dramatically within species.

Hopane-type saponins from Glinus lotoides Linn.

Seven hopane-type saponins were isolated from the methanol extract of Glinus lotoides. Six of them were identified as novel compounds and designated as lotoideside A [3-O-beta-D-xylopyranosyl (1-->2)-alpha-L-rhamnopyranosyl-6 alpha-O-beta-D-xylopyranosyl-22-beta-O-beta-D-glucopyranosyl-16 beta-hydroxy hopane (1)], lotoideside B [3-O-beta-D-xylopyranosyl (1-->2)-alpha-L-rhamnopyranosyl-22-beta-O-beta-D-glucopyranosyl-6 alpha,16 beta-dihydroxyhopane (2)], lotoideside C [3-OD-xylopyranosyl-6 alpha-O-beta-D-xylopyranosyl-16 beta-O-beta-D-xylopyranosyl-22 beta-hydroxyhopane (3)], lotoideside D [3-O-beta-D-xylopyranosyl-16 beta-O-alpha-L-arabinopyranosyl-6 alpha,22-beta-dihydroxyhopane (4)], lotoideside E [3-O-beta-D-xylopyranosyl-6 alpha-O-beta-D-xylopyranosyl-16 beta,22-beta-dihydroxyhopane (5)], and lotoideside F [3-O-beta-D-xylopyranosyl-22-beta-O-beta-D-glucopyranosyl-16 beta-hydroxyhopan-6-one (6)]. The known compound succulentoside B (7) was also encountered. Their structures were elucidated on the basis of one-and two-dimensional NMR spectroscopic techniques, ESIMS and chemical evidences.