Anxiolytic potential of ursolic acid derivative-a stearoyl glucoside isolated from Lantana camara L. (verbanaceae)

ObjectiveTo investigate the anxiolytic activity of newly isolated compound by our lab called ursolic acid stearoyl glucoside (UASG) from the leaves ofLantana camara (L. camara).MethodsColumn chromatography was used to isolate UASG. Anxiolytic potential was experimentally proved and demonstrated through Elevated plus-maze, Open field and light and dark test.ResultsThe UASG showed marked increased in time spent (%) and number of frequent movements made by animals in open arm of elevated plus-maze apparatus. In light and dark model, UASG produced marked increase in time spent by animal, number of crossing and reduced duration of immobility in light box.ConclusionsUASG showed significant increase in number of rearing, assisted rearing and number of square crossed in open field established test model. UASG showed its anxiolytic effect in dose dependent manner.     

Anti-hepatoma activity and mechanism of ursolic acid and its derivatives isolated from Aralia decaisneana

AIM:To investigate the anti-tumor activity of ursolic acid(UA)and its derivatives isolated from Aralia decaisneanaon hepatocellular carcinoma both in vitro and in vivo.METHODS:In vivo cytotoxicity was first screened by3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide(MTT)assay.Morphological observation,DNAladder,flow cytometry analysis,Western blot and realtime PCR were employed to elucidate the cytotoxicmechanism of UA.Implanted mouse hepatoma H_(22)wasused to evaluate the growth inhibitory effect of UA invivo.RESULTS:UA could significantly inhibit the proliferationof HepG2 and its drug-resistance strain,R-HepG2 cells,but had no inhibitory effect on primarily cultured normalmouse hepatocytes whereas all the six derivativesof UA could not inhibit the growth of all tested celllines.Further study on mechanism demonstratedthat apoptosis and G_0/G_1 arrest were involved inthe cytotoxicity and cleavage of poly-(ADP-ribose)-polymerase(PARP).Downregulation of cyclooxygenase-2(COX-2)protein and upregulation of heat shock protein(HSP)105 mRNA correlated to the apoptosis of HepG2cells treated with UA.In addition,UA also could inhibitthe growth of H_(22)hepatoma in vivo.CONCLUSION:UA is a promising anti-tumor agent,butfurther work needs to be done to improve its solubility.     

Proteins with insulinlike activity isolated from oyster (Ostrea edulis L.) hepatopancreas

Three radioactive protein peaks, I, II, and III, with insulinlike activity, have been obtained from oyster hepatopancreas by chromatography in Biogel-P30 columns. The molecular weight values for the peaks were 14,000, 11,000, and 7000, respectively. Trypsin treatment transformed peak II into peak I. Peak I was transformed into peak III by incubation at pH 9. The insulinlike activity of the three peaks was decreased by treatment with cysteine. The presented results suggest that peaks I, II, and III contain insulin dimer, proinsulin, and insulin, respectively.     

Mosquito larvicidal constituents from Lantana viburnoides sp viburnoides var kisi (A. rich) Verdc (Verbenaceae)

BACKGROUND & OBJECTIVES: Lantana viburnoides sp viburnoides var kisi is used in Tanzania ethnobotanically to repel mosquitoes as well as in traditional medicine for stomach ache relief. Bioassay-guided fractionation and subtraction bioassays of the dichloromethane extract of the root barks were carried out in order to identify the bioactive components for controlling Anopheles gambiae s.s. mosquito larvae. METHODS: Twenty late III or early IV instar larvae of An. gambiae s.s. were exposed to various concentrations of the plant extracts, fractions, blends and pure compounds, and were assayed in the laboratory by using the protocol of WHO 1996. Mean mortalities were compared using Dunnett's test (p < 0.05) and lethal concentration calculated by Lackfit Inversel of the SAS programme. RESULTS: The crude extract (LC50 = 7.70 ppm in 72 h) and fractions exhibited different level of mosquito larvicidal activity with subtraction of some fractions resulting in activity enhancement. The active fractions contained furanonaphthaquinones regio-isomers (LC50 = 5.48-5.70 ppm in 72 h) and the lantadene triterpenoid camaric acid (LC50 = 6.19 ppm in 72 h) as active principles while the lupane triterpenoid betulinic acid (LC50 < 10 ppm in 72 h) was obtained from the least active fraction. INTERPRETATION & CONCLUSION: Crude extracts and some fractions had higher or comparable larvicidal activity to the pure compounds. These results demonstrate that L. viburnoides sp viburnoides var kisi extracts may serve as larvicides for managing various mosquito habitats even in their semi-purified form. The isolated compounds can be used as distinct markers in the active extracts or plant materials belonging to the genus Lantana.     

Potential carcinogenic and inhibitory activity of compounds isolated from Lilium candidum L

This paper deals with determination of potential carcinogenic and inhibitory activity of 10 compounds isolated from the ethanolic extract of Lilium candidum L. perfonned by DC polarography. The series of investigated compounds consisted of two spirostanol saponins, two pyroline derivatives, jatropham and its glucoside, flavonol kaempferol, 2-fenylethyl-alpha-L-arabinopyranosyl-(1-->6)-beta-D-glucopyranoside, 2-phenylethylpalmitate and methylsuccinic acid. Carcinogenic, resp. mutagenic activity data for these compounds, with the exception of kaempferol, are not available in scientific literature. All tested compounds were reduced in N,N-dimethylformamide in one or two well defined steps. They also fonned a reversible complex with alpha-lipoic acid, a compound serving as an indicator of carcinogenic activity. The marginal diffuse current values of these complexes served as a criterion of a very low potential carcinogenicity. The inhibitory activity of isolates were studied in the presence of 12-O-tetradecanoylphorbol-13-acetate, a specific tumor promoter for an epidermal carcinogenesis, and in the presence of a polyaromatic substance 7,12-dimetylbenz(a)anthracene known as a carcinogen. The spirostanol saponins possessed the highest inhibitory activity (> 70%) and jatropham (66%). The inhibitory activity of jatropham glucoside was significantly lower (49%). Practically zero inhibitory activity was measured for the 2-phenylethylpalmitate and methylsuccinic acid.     

Antileukemia (L1210) activity and toxicity of cis-dichlorodiammineplatinum(II) analogs.

cis-Dichlorodiammineplatinum(II) (cis-platinum) and 40 of its analogs were evaluated for antitumor activity in BDF1 mice implanted ip with 10(6) L1210 leukemia cells. Several of these analogs were also evaluated for their ability to cause elevation in BUN and to produce leukopenia in normal BDF1 mice. In 11 experiments, cis-platinum given on Day 1 or Days 1--9 after implant produced T/C (treated/control) values between 164% and 214% and 157% and 285% respectively. On the basis of single experiments, 13 analogs were judged to be comparable to cis-platinum in that they produced T/C values greater than or equal to 167% after a single dose or greater than or equal to 200% after daily doses for 9 days. These active compounds included various substituted amine derivatives of dichloroplatinum(II), malonatoplatinum(II), aquasulfatoplatinum(II), and chloroacetatoplatinum(II) and derivatives of dihydroxydichloroplatinum(IV). Twelve of these active analogs and cis-platinum were evaluated for toxicity at doses ranging from near the optimal therapeutic dose to greater than or equal to the LD50. Only five of the 12 analogs and cis-platinum caused an increase in BUN to greater than 30 mg/100 ml, while eight of the analogs produced leukopenia comparable in incidence and severity to that produced by cis-platinum.     

Anti-trypanosomal activity of pentacyclic triterpenes isolated from Austroplenckia populnea (Celastraceae)

Four pentacyclic triterpenes isolated from Austroplenckia populnea and four compounds of known anti T. cruzi or anti-malarial activity were tested. Of those triterpenes tested 20alpha-hydroxy-tingenone showed high activity, epikatonic acid was less active, while populnilic and populninic acids were inactive against the trypanosome of the subgenus Schizotrypanum tested. Benzonidazole, nifurtimox, ketoconazole and primaquine presented a remarkable dose-dependent inhibitory effect reaching practically to a total growth inhibition of the parasite at the end of incubation time. The trypanosome tested appear to be a suitable model for preliminary screen for anti T. (S.) cruzi compounds.     

Antimicrobial activity and composition of the volatiles of Cinnamomum tamala Nees. and Murraya koenigii (L.) Spreng. from Uttarakhand (India)

ObjectiveTo examine the composition of Cinnamomum tamala and Murraya koenigii essential oils and their antimicrobial activities against nine microbial strains.MethodsEssential oils were obtained by hydrodistillation from the leaves of two spice trees and were analyzed by GC and GC/MS. The oils were also tested for their antimicrobial activity using broth micro dilution method.ResultsCinnamaldehyde (37.85%) and cis-linalool oxide (29.99%) were the main components characterized in the oil of C. tamala, whereas α-pinene (39.93%), sabinene (13.31%) and trans-caryophyllene (9.02%) detected as the major constituents in M. koenigii oil. C. tamala oil exhibited significant antifungal activity and satisfactory antibacterial activity, while lesser antimicrobial activity was observed in M. koenigii oil.ConclusionsThe present study suggested that C. tamala oil was more effective against bacterial and fungal strains as compared with M. koenghii oil.     

Anticancer activity test of ethyl acetate extract of endophytic fungi isolated from soursop leaf(Annona muricata L.)

Objective: To analyze anticancer activity of an ethyl acetate extract of endophytic fungi isolated from soursop leaf(Annona muricata L.). Methods: Anticancer activity of fungal extracts was determined by observing its toxicity against MCF-7(Michigan Cancer Foundation-7) cells in vitro by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay method. At an extract concentration of 100 μg/m L, 4 isolates out of 12 showed high activity against the cancer cell growth. The four isolates were then selected for further IC50 determination, by measuring the inhibition of cancer cell proliferation at extract concentration of 25 μg/m L, 50 μg/m L, 100 μg/m L, 200 μg/m L and 400 μg/m L. Results: Results showed that isolate Sir-G5 had the highest anticancer activity with an IC50 of 19.20 μg/m L. The best isolates were screened again using a normal cell(Chang cells) to determine its toxicity against normal cells. Results indicated that the extracts do not affect the proliferation of normal cells. Molecular identification showed that the fungal isolate Sir-G5 has a close relationship with Phomopsis sp. Conclusions: The endophytic fungi isolated from soursop leaf has the potential to be used as a source of anticancer agents.     

Production of erythroid burst-promoting activity (BPA) and granulocyte-monocyte colony-stimulating activity (GM-CSA) by isolated human T-lymphocyte subpopulations

T-lymphocytes and monocytes are prominent among the classes of normal human cells that have been implicated in the production of the hemopoietic growth factors granulocyte-macrophage colony-stimulating activity (GM-CSA) and erythroid burst-promoting activity (BPA). To investigate the nature of the cooperativity that occurs during the elaboration of these growth factors by activated T-lymphocytes and monocytes in vitro, and to define the subsets of T cells involved in this response, we studied the production of GM-CSA and BPA by populations of T-lymphocytes isolated by fluorescence-activated cell sorting, using the monoclonal antibodies OKT3, OKT4, and OKT8. When OKT3+, OKT4+, or OKT8+ cells were incubated for five days in liquid suspension cultures, their production of GM-CSA and BPA was undetectably low. When 5% autologous monocytes were added to the cultures, no increase in the secretion of either of these classes of growth factors was noted. In the presence of concanavalin A (Con A), measurable quantities of both GM-CSA and BPA were elaborated by all three populations of T cells in the absence of monocytes; however, when autologous monocytes were added to the Con A-stimulated T cells, the secretion of both GM-CSA and BPA was markedly enhanced. In addition, we found that supernates of unfractionated T cells incubated with Con A contained not only GM-CSA and BPA but also a potent inhibitor(s) of BPA that could be demonstrated by dilution of the media and removed by gel filtration. In contrast, no inhibitor of GM-CSA was found. By molecular sieve chromatography of the supernates, GM-CSA and BPA coeluted as a single peak. However, the two biologic activities could be separated on the basis of heat stability, since GM-CSA proved to be heat labile whereas BPA did not. Our data indicate that GM-CSA and BPA derived from human T cells are similar in their apparent molecular weights and in the pattern of their production in suspension cultures in response to lectin stimulation. The secretion of both GM-CSA and BPA by Con A-stimulated T cells is facilitated by the presence of autologous monocytes, and is not restricted to either the OKT4- or the OKT8-defined subset.     

In vitro activity of methylrosaniline chloride (gentian violet) as disinfectant against Candida spp. and Trichosporon spp. isolated from blood samples

OBJECTIVE: Methylrosaniline Chloride (MRC) is recognized as a disinfectant, but recently is rarely used in the clinic, because of its cytotoxicity when used continuously with conventional concentrations (1% MRC). We have reported the antibacterial activity of MRC with lower concentration against Methicillin-resistant Staphylococcus aureus (MRSA). In this study, we evaluated the antifungal activity of MRC with lower concentrations. MATERIAL AND METHODS: Antifungal activities of MRC against Candida spp. and Trichosporon spp. were tested. All strains tested were isolated from 106 blood or intravenous catheter samples at Juntendo University Hospital from 1995 to 2004. Minimum inhibitory concentrations against fungi were assayed by agar dilution, under both aerobic and anaerobic conditions. RESULTS: A 0.01% or less concentration of MRC solutions showed marked antifungal activity against Candida spp. and Trichosporon spp. under aerobic or anaerobic conditions. CONCLUSION: A 0.01% or less concentration of MRC should be reevaluated for the control of fungal infection and MRSA infection control.     

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

ObjectiveTo assess antifungal activity of Ocimum sanctum leaves against dermatophytic fungi.MethodsAntifungal 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.ResultsOcimum 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.ConclusionsOcimum sanctum has antifungal activity, and the leaf extracts may be a useful source for dermatophytic infections.     

Effect of niloticin,a protolimonoid isolated from Limonia acidissima L. (Rutaceae) on the immature stages of dengue vector Aedes aegypti L. (Diptera: Culicidae)

The aim of the present study was to evaluate the mosquitocidal activity of fractions and a compound niloticin from the hexane extract of Limonia acidissima L. leaves on eggs, larvae and pupae of Aedes aegypti L. (Diptera: Culicidae). In these bioassays, the eggs, larvae and pupae were exposed to concentrations of 2.5, 5.0, 7.5 and 10.0 ppm for fractions and 0.5, 1.0, 1.5 and 2.0 ppm for compound. After 24 h, the mortality was assessed and the LC₅₀ and LC₉₀ values were calculated for larvae and pupae. Per cent ovicidal activity was calculated for eggs after 120 h post treatment. Among the sixteen fractions screened, fraction 8 from the hexane extract of L. acidissima generated good mosquitocidal activity against Ae. aegypti. The LC₅₀ and LC₉₀ values of fraction 8 were 4.11, 8.04 ppm against Ae. aegypti larvae and 4.19, 8.10 ppm against Ae. aegypti pupae, respectively. Further, the isolated compound, niloticin recorded strong larvicidal and pupicidal activities. The 2 ppm concentration of niloticin showed 100% larvicidal and pupicidal activities in 24 h. The LC₅₀ and LC₉₀ values of niloticin on Ae. aegypti larvae were 0.44, 1.17 ppm and on pupae were 0.62, 1.45 ppm, respectively. Niloticin presented 83.2% ovicidal activity at 2 ppm concentration after 120 h post treatment and niloticin exhibited significant growth disruption and morphological deformities at sub lethal concentrations against Ae. aegypti. The structure of the isolated compound was identified on the basis of single XRD and spectral data (¹H NMR and ¹³C NMR) and compared with literature spectral data. The results indicate that niloticin could be used as a potential natural mosquitocide.     

(-)-baclofen and gamma-aminobutyric acid inhibit calcium currents in isolated retinal ganglion cells.

Of the various synaptic inputs known to converge upon retinal ganglion cells, the major inhibitory inputs are thought to be GABAergic. Although gamma-aminobutyric acid (GABA) is known to activate anion-selective ion channels in retinal ganglion cells, we have tested the possibility that GABA can also modulate cationic conductances in these cells, as seen in other central and peripheral neurons. Specifically, we have made whole-cell patch-clamp recordings to test whether voltage-gated calcium currents in isolated goldfish retinal ganglion cells are sensitive to GABAB receptor ligands. (-)-Baclofen and GABA inhibited calcium currents activated by moderately long depolarizations and, during large depolarizations (e.g., to 0 mV), also appeared to accelerate the rate of current decay. The calcium current inhibition induced by (-)-baclofen and GABA was not prevented by 2-hydroxysaclofen, phaclofen, or bicuculline, even though bicuculline suppressed a GABA-activated conductance in these cells. These results demonstrate the presence of baclofen- and GABA-sensitive calcium currents in vertebrate retinal ganglion cells as well as the coexistence of GABAA and GABAB receptors in individual retinal ganglion cells.     

Cardiovascular effects of mollic acid glucoside, a 1alpha-hydroxycycloartenoid saponin extractive from Combretum molle R Br ex G Don (Combretaceae) leaf

The cardiovascular effects of mollic acid glucoside (MAG), a 1alpha-hydroxycycloartenoid saponin extractive from Combretum molle R Br ex G Don (Combretaceae) leaf, have been investigated in some experimental animal paradigms. The plant extract (MAG, 5-80 microg/ml) produced concentration-dependent, significant (p < 0.05-0.001) negative inotropic and negative chronotropic effects on guinea pig isolated electrically driven left, and spontaneously beating right atrial muscle preparations, respectively. MAG also significantly reduced (p < 0.05-0.001) or abolished, in a concentration-dependent manner, the rhythmic, spontaneous contractions of portal veins isolated from healthy, normal Wistar rats. Like acetylcholine (ACh, 10-(8)-10-(5) M), the plant extract (5-80 microg/ml) produced concentration-related relaxations of rat isolated endothelium-containing thoracic aortic rings pre-contracted with noradrenaline (NA, 10-10(-10)-(5) M). The vasorelaxant effects of MAG in the aortic rings were markedly inhibited or annulled by N(G)-nitro-L-arginine methyl ester (L-NAME, 10(-5) M), a nitric oxide synthase inhibitor. Furthermore, MAG (2.5-40 mg/kg iv) caused dose-related, transient but significant reductions (p < 0.05-0.001) in the systemic arterial blood pressures and heart rates of anaesthetised normotensive and hypertensive rats. The results of this laboratory animal study indicate that MAG caused bradycardia, vasorelaxation and hypotension in the mammalian experimental models used. The vasorelaxant action of MAG was endothelium dependent, and was therefore possibly dependent on the synthesis and release of nitric oxide (NO). The findings of this study suggest that Combretum molle leaf may be used as a natural supplementary remedy in essential hypertension and in certain cases of cardiac dysfunctions in rural African communities.     

瑞香狼毒药效组分对裸鼠肝癌移植瘤的抑制及其机制

目的:评价瑞香狼毒药效组分抑制人肝癌细胞株BEL-7402裸鼠皮下移植瘤作用的强度,并探讨其机制.方法:建立人肝癌BEL-7402裸小鼠皮下移植瘤模型研究瑞香狼毒药效组分Zp1111抗肿瘤作用与对荷瘤裸鼠体质量与免疫器官的影响,免疫组织化学法检测用药前后BEL-7402皮下移植瘤组织Bcl-2与Bax阳性表达的变化.结果:Zp1111对裸小鼠皮下移植瘤BEL-7402具有较强的抑制作用,187.5mg/kg、250mg/kg(折合成生药量)剂量时对移植瘤的相对肿瘤增殖率T/C小于60%;该组分能显著上调BEL-7402移植瘤组织Bax基因的表达,但对Bcl-2的表达影响较弱.结论:瑞香狼毒药效组分在体内对人肝癌细胞BEL-7402裸鼠移植瘤具有较强的抑制作用并呈现一定的剂量依赖关系,该组分具有一定的诱导凋亡作用,可能与上调Bax有关.     

Translation of messenger ribonucleic acid from isolated pancreatic islets and human insulinomas.

RNA preparations from isolated rat pancreatic islets and from human insulinomas were injected into oocytes of Xenopus laevis which were then incubated with [3H]leucine. Acid-ethanol extracts of the oocytes were immunoprecipitated with anti-insulin serum using the double antibody technique. Sodium dodecyl sulfate disc gel electrophoresis of the immunoprecipitates showed the presence of an insulin-displaceable immunoreactive material with a molecular weight of about 18,000 in extracts from oocytes injected with RNA of 9-11 S. This immunoreactive product was not detected in extracts from oocytes injected with buffer or 4-8S RNA or RNA heavier than 11 S. These observations suggest the involvement of a precursor larger than proinsulin in the biosynthesis of insulin.