Release of cytokines by a fermented lectin-1 (ML-1) free mistletoe extract reflects differences in the reactivity of PBMC in healthy and allergic individuals and tumour patients

Objective: Mistletoe extracts are used in adjuvant cancer treatment, but little is known concerning their mode of action. There is, however, evidence that antigens in these extracts may stimulate cells of the immune system, thereby modifying the altered immunological reactivity in tumour patients. Methods: In order to find out whether the postulated immunomodulatory properties of mistletoe extracts are mediated by cytokines, a spectrum of different cytokines was analysed in the supernatants of peripheral blood mononuclear cells (PBMC) from healthy (n = 23) and allergic (n = 16) individuals after stimulation with the fermented mistletoe lectin-1 (ML-1) free mistletoe extract Iscador Pini (IP) in vitro, and their cytokine patterns were compared to those from tumour patients with either breast cancer (n = 20) or colorectal tumours (n = 22). Results: PBMC from healthy and allergic individuals produced high levels of TNF-α and IL-6 and to a lesser extent Th1- and Th2-related cytokines. This finding was in contrast to data obtained in tumour patients. Thus, the concentration of TNF-α was significantly lower in the cell cultures from breast cancer patients than in controls, and patients with colorectal tumours released IFN-γ/IL-2 (5%) in the supernatants significantly less frequently than PBMC from healthy controls (26%). Similar results were obtained when the Th1- and monocyte/macrophage-related cytokines were analysed in the unstimulated cell cultures. Conclusions: These in vitro studies provide evidence that there is a reduced immunological reactivity to the fermented ML-1 free mistletoe extract in tumour patients and may give some clues as to how mistletoe-derived antigens could act on immune cells involved in the tumour defence. Received: 18 March 1996/Accepted in revised form: 18 July 1996     

Human cancer cells exhibit in vitro individual receptiveness towards different mistletoe extracts

In vitro cytotoxic effects of three aqueous mistletoe extracts on cell physiology against different human tumor cell lines and primary cancer cells were investigated in order to compare the receptiveness of different cancer cells against different mistletoe products. Therefore cell proliferation (BrdU-incorporation assay), mitochondrial activity (MTT-testing) and necrotic cell toxicity (LDH assay) were assayed over serial dilutions of the test products. Data obtained with HELA-S3, MOLT-4, MFM-223, COR-L51, KPL-1 and VM-CUB1 tumor cell lines and Iscador M (20 mg/ml), Iscador Q (20 mg/ml) and Abnobaviscum Fraxini -2 (20 mg/ml) indicated significant growth-inhibition of all cell lines, but also different cell susceptibilities against the different extracts. These variations were not only monitored on established cell lines but also on primary mamma carcinoma cells from surgical resectates. Concerning cell proliferation and mitochondrial activity Abnobaviscum Fraxini exhibits stronger inhibitory effects compared to products from the Iscador series. In case the evaluation was standardized on the active contents of VAA-I within the different products, the Iscador extracts possess higher cytotoxic activity. Pure viscotoxins and mistletoe lectins exhibited less effects than the extracts. The simultaneous presence of pure mistletoe lectins and mistletoe polysaccharides diminished the VAA-mediated cytotoxic effects. The presence of fetal calf serum (FCS) in cultivation media during in vitro testing diminished the cytotoxic effects of mistletoe extracts. It was shown that in vivo application of mistletoe preparations led to the formation of antibodies against unknown compounds of the extracts, diminishing the cytotoxic effect.     

Absence of tumor growth stimulation in a panel of 16 human tumor cell lines by mistletoe extracts in vitro

Extracts of Viscum album (mistletoe) are widely used as complementary cancer therapies in Europe. The mistletoe lectins have been identified as the main active principle of mistletoe extracts. They have been shown to exhibit cytotoxic effects as well as immunomodulatory activities. The latter is exemplified by induction of cytokine secretion and increased activity of natural killer cells. Recent reports, however, indicated possible tumor growth stimulation by mistletoe extracts. Therefore, the three aqueous mistletoe extracts (Iscador M special, Iscador Qu special and Iscador P) were evaluated for antiproliferative and/or stimulatory effects in a panel of 16 human tumor cell lines in vitro using a cellular proliferation assay. The results show no evidence of stimulation of tumor growth by any of the three Iscador preparations, comprising central nervous system, gastric, non-small cell lung, mammary, prostate, renal and uterine cancer cell lines, as well as cell lines from hematological malignancies and melanomas. On the contrary, Iscador preparations containing a high lectin concentration (Iscador M special and Iscador Qu special) showed antitumor activity in the mammary cancer cell line MAXF 401NL at the 15 microg/ml dose level with a more than 70% growth inhibition compared to untreated control cells. In addition, a slight antitumor activity (growth inhibition 30-70%) was found in three tumor cell lines for Iscador M special and in seven tumor cell lines for Iscador Qu special, respectively. Iscador P, which contains no mistletoe lectin I, showed no antiproliferative activity.     

Prospective controlled cohort studies on long-term therapy of ovairian cancer patients with mistletoe (Viscum album L.) extracts iscador

BACKGROUND: Mistletoe extracts such as Iscador are commonly used as complementary/anthroposophic medications for many cancer indications, particularly for solid cancers. The efficacy of this complementary therapy is still controversial. OBJECTIVE: Does long-term therapy with mistletoe extracts Iscador show any effect on survival and psychosomatic self-regulation of patients with ovarian cancer? Patients and methods: Prospective recruitment and long-term follow-up in controlled cohort studies. (1) Two randomized matched-pair studies: OvarRand (ovarian cancer patients without distant metastases; 21 pairs) and OvarMetRand (ovarian cancer patients with distant metastases; 20 pairs); patients having no mistletoe therapy were matched for prognostic factors. By paired random allocation, one of the patients of each pair was suggested therapy with mistletoe extracts Iscador to be applied by her attending physician. (2) Two non-randomized matched-pair studies: Ovar (ovarian cancer patients without distant metastases; 75 pairs) and OvarRand (ovarian cancer patients with distant metastases; 62 pairs); patients that already received therapy with mistletoe extracts Iscador were matched by the same criteria to control patients without therapy with mistletoe extracts Iscador. RESULTS: For overall survival in the randomized studies, the effect in favor of therapy with mistletoe extracts Iscador was significant in OvarMetRand but not in OvarRand; hazard ratio estimate and 95% confidence interval: 0.40 (0.15, 1.03) and 0.33 (0.12, 0.92), respectively. In the non-randomized studies Ovar and OvarMet, the results adjusted for relevant prognostic variables were 0.47 (0.31, 0.69) and 0.62 (0.37, 1.05). Psychosomatic self-regulation in the Iscador group increases significantly within 12 months on a scale from 1 to 6 compared with the control group in the randomized study OvarRand as well as in the non-randomized study Ovar on patients with ovarian cancer without distant metastases; estimate of the median difference and 95% confidence interval: 0.58 (0.30, 0.90) and 0.30 (0.05, 0.65), respectively. CONCLUSION: Mistletoe extracts Iscador might have the effect of prolonging overall survival of ovarian cancer patients. In the short term, psychosomatic self-regulation increases more markedly under Iscador therapy than under conventional therapy alone.     

金荞麦提取物体外抗流感病毒作用研究

目的 研究金荞麦提取物的体外抗流感病毒作用。方法 采用血凝试验,考察金荞麦提取物对鸡胚内流感病毒增殖的抑制作用;采用MTT法检测细胞活性,考察金荞麦提取物对流感感染细胞活性的影响。结果 与病毒对照组比较,金荞麦提取物高剂量组（10 mg·mL^-1）、中剂量组（5 mg·mL^-1）均能显著降低鸡胚尿囊液的血凝滴度（P<0.01）,而低剂量组（2.5 mg·mL^-1）则无显著性差异;在0.63~2.50 mg·mL^-1浓度内,金荞麦提取物各剂量组流感感染细胞的活性均明显高于病毒对照组,组间均存在显著性差异（P<0.01）,且病毒抑制率随着药物浓度的增大而增高,呈一定的量效关系,其中金荞麦提取物最大无毒浓度（2.5 mg·mL^-1）对流感病毒的直接抑制率达66.12%。结论 金荞麦提取物具有一定的体外抗流感病毒作用。     

Chemopreventive potential of Epoxy clerodane diterpene from <Emphasis Type="Italic">Tinospora cordifolia</Emphasis> against diethylnitrosamine-induced hepatocellular carcinoma

Summary  Medicinal plants are a promising source for identification of lead molecules for cancer therapy. In our continuous search to discover bioactive compounds from natural products, we isolated (5R, 10R)-4R, 8R-dihydroxy-2S, 3R:15, 16-diepoxycleroda-13(16), 17, 12S:18,1S-dilactone (ECD), a diterpenoid from Tinospora cordifolia and studied its chemopreventive potential in diethylnitrosamine (DEN) induced hepatocellular carcinoma (HCC) rats. Fifty male Wistar rats were divided into five groups. Group I served as normal control. Group II–IV were given DEN (0.01% in drinking water) for twenty weeks. In addition, Group III (preventive treatment) received ECD (10 mg/kg body weight) throughout the study. Group IV (curative treatment) received ECD (10 mg/kg body weight) for the last 8 weeks. Group V received ECD alone (10 mg/kg body weight) throughout the experimental period. At the end of the experimental period all the animals were sacrificed and analyzed for biochemical end points to assess the effect of ECD treatment in DEN induced HCC. The animals treated with DEN showed a decrease in the activities of antioxidant (SOD, CAT) and detoxification enzymes (GSH, GPx) with increase in the activities of the hepatic markers (SGOT, SGPT, LDH). Treatment of ECD in both preventive and curative DEN induced animals increased the level of antioxidants and detoxification enzymes, and decreased serum transaminase level and hepatic marker enzymes to near normal. Histopathological and nodular incidence also confirmed that ECD remarkably reduced tumor incidence and reversed damaged hepatocytes to normal. Our findings confirm that ECD exhibits preventive effect against chemically induced HCC in rats. ECD can be a potent chemopreventive drug for HCC.     

Serenoa repens associated with Urtica dioica (ProstaMEV) and curcumin and quercitin (FlogMEV) extracts are able to improve the efficacy of prulifloxacin in bacterial prostatitis patients: results from a prospective randomised study

We report the results of a prospective randomised study to evaluate the therapeutic effect of Serenoa repens, Urtica dioica (ProstaMEV^®), quercitin and curcumin (FlogMEV^®) extracts associated with prulifloxacin in patients affected by chronic bacterial prostatitis (CBP). From a whole population of 284 patients, 143 patients affected by CBP [National Institutes of Health (NIH) class II prostatitis] were enrolled. All patients received prulifloxacin 600 mg daily for 14 days, in accordance with antibiogram results. Patients were split into two groups: Group A received prulifloxacin associated with ProstaMEV^® and FlogMEV^®; Group B received only antibiotic therapy. Microbiological and clinical efficacies were tested by two follow-up visits at 1 month and 6 months, respectively. Quality of life (QoL) was measured using the NIH Chronic Prostatitis Symptom Index (CPSI) and International Prostatic Symptom Score (IPSS) questionnaires. Group A comprised 106 patients and Group B comprised 37 patients. One month after treatment, 89.6% of patients who had received prulifloxacin associated with ProstaMEV^® and FlogMEV^® did not report any symptoms related to CBP, whilst only 27% of patients who received antibiotic therapy alone were recurrence-free (P < 0.0001). Significant differences were found between groups in terms of symptoms and QoL (P < 0.0001 for both). Six months after treatment, no patients in Group A had recurrence of disease whilst two patients in Group B did. Questionnaire results demonstrated statistically significant differences between groups (all P < 0.001). The association of S. repens, U. dioica (ProstaMEV^®), quercitin and curcumin (FlogMEV^®) extracts is able to improve the clinical efficacy of prulifloxacin in patients affected by CBP.     

Chronic Effects of an Insecticide, Fenobucarb, on the Larvae of Two Mayflies, Epeorus latifolium and Baetis thermicus, in Model Streams

Effects of the insecticide fenobucarb on 2 mayfly species, Epeorus latifolium and Baetis thermicus, were examined in indoor model streams. Aqueous concentrations of fenobucarb residues in the model streams were 69–71% of its nominal level until 24 h, except at low concentrations (less than 2 g l^–1 in the water) in the water. Chronic effects of fenobucarb on the growth and emergence of Ephemeropteran larvae were examined over 2 months. For concentrations of 1 and 2 g l^–1, the numbers of individuals of the third larval growth stage (average head width 3.0 mm) gradually decreased up to 20 days after the applications; the number of emergent individuals also remained 20 to 25 by the end of the experiment. The number of individuals to emerge at 1 to 2 g l^–1 fenobucarb was restricted in the long-term experiment. These results suggest that fenobucarb affects the emergence of this species through disruption of the endocrine system. On the other hand, the number of emergent individuals of B. thermicus at low concentrations of fenobucarb was not significantly less than that in the control.     

Anti-staphylococcal activity of ent-kaurane-type diterpenoids from Croton tonkinensis

Ent-kaurane-type diterpenpoids 1–11, isolated from the dried leaves of the endemic Vietnamese medicinal plant Croton tonkinensis Gagnep. (Euphorbiaceae), were evaluated for inhibitory activity against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) strains. The most active diterpenoids, 2, 3, and 8, exhibited minimum inhibitory concentrations (MICs) of 32, 500, and 125 g/ml, respectively, against MRSA strains.     

Neurotoxicity and pharmacology of Lathyrus sativus extracts of high- and low-toxicity strains

Neurolathyrism is characterized by spastic paraparesis of the legs. It is caused by overconsumption of grass pea (Lathyrus sativus L.; Leguminosae). We studied toxicity of extracts of L. sativus seeds from two different areas—Bangladesh and Canada—toward rat primary neuron/glia culture. Both extracts showed acute neurotoxicity within 24 h when the 75% ethanol extracts were added to the neuron/glia culture. Fractionation of the extracts showed that the water-soluble fraction accounted for ca. 75–84% of total toxicity in which 3-N-oxalyl-l-2,3-diaminopropanoic acid (l--ODAP) was present at the highest concentration. Toxicity of the water-soluble fraction obtained from Bangladeshi seeds was significantly higher than that obtained from Canada. Effects of these fractions were reversed almost completely by 1,2,3,4-tetrahydro-6-nitro-2,3-dioxobenzo[f]quinoxaline-7-sulfonamide (NBQX), an antagonist of AMPA-receptor. They were partially reversed by group I metabotropic glutamate receptor antagonists (RS)-1-aminoindan-1,5-dicarboxylic acid, or (S)--methyl-4-carboxyphenyl-glycine [(S)-MCPG]. Nitric oxide synthase inhibitor N^G-nitro-l-arginine methyl ester (l-NAME) strongly decreased the extracts toxicity. These data show that the neurotoxicity of grass pea seeds is attributable to l--ODAP, the toxicity of which is mediated by collective effects of l--ODAP on the AMPA-type receptor, metabotropic glutamate receptors, and NO production.     

Mechanisms of p,p′-DDE-induced apoptosis in human peripheral blood mononuclear cells

p,p′-Dichlorodiphenyldichloroethylene (DDE), the most stable metabolite of organochlorine insecticide p,p′-dichlorodiphenyltrichloroethane (DDT), has been detected in human populations living in malaria-endemic areas of México where this insecticide was used. DDE induces apoptosis in peripheral blood mononuclear cells (PMBC); however, the molecular mechanism of cell death induced by this compound is poorly understood. In the present study, PBMC isolated from healthy individuals (not exposed to DDE) were incubated in the presence of increasing concentrations of p,p′-DDE (0–80 μg/ml) over time. When PBMC were treated with low p,p′-DDE concentration (10 μg/ml) an antioxidant response and biomarkers of inflammation were induced, indicating a pro-inflammatory state. Moreover, when PBMC were treated with high p,p′-DDE concentration (80 μg/ml) several apoptotic biochemical events were triggered, such as activation of caspase-8, Bid, caspase-9 and caspase-3, as well as degradation of PARP and ubiquitination. The results described in this study show a possible inflammatory condition and the involvement of both extrinsic and intrinsic pathways in the induction of apoptosis in DDE-treated PBMC.     

HPLC profile and antihyperglycemic effect of ethanol extracts of <Emphasis Type="Italic">Andrographis paniculata</Emphasis> in normal and streptozotocin-induced diabetic rats

Andrographis paniculata is a Malaysian traditional herb for treatment of fever-causing diseases and diabetes. The aim of the present study was to examine the antihyperglycemic effects of different extracts (95, 50, and 20% ethanol extracts and water extracts) of A. paniculata in normal and streptozotocin-induced diabetic rats. Only the treatment with 20% ethanol extract (at 1 g/kg for 7 h, p.o.) significantly (P < 0.01) reduced the rise in blood glucose levels in glucose-loaded rats. None of the different extracts (at 1 g/kg for 7 h, p.o.) significantly (P > 0.05) altered the blood glucose levels in normal and streptozotocin-induced diabetic rats as compared to the control group. However, administration of the 50 and 95% ethanol extracts (at 1 g/kg for 14 days, p.o.) significantly (P < 0.05) reduced fasting blood glucose and insulin levels in normal and diabetic rats compared to the pre-treatment level. HPLC profiles of the extracts were developed using andrographolide (AP) as a marker. The amounts of AP in 95, 50, and 20% ethanol extracts and water extracts were 25.8, 19.4, 2.0, and 0.8 mg/g of dry weight, respectively.     

Novel GHB-derived natural products from European mistletoe (Viscum album)

Abstract

Context: The European white-berry mistletoe [Viscum album L. (Loranthaceae)] is among the oldest known medicinal plants. At present the most important application of mistletoe extracts is in the treatment of cancer. However, natural products specific to mistletoe have rarely been encountered in the current literature.

Objective: To discover novel natural products specific to European mistletoe.

Materials and methods: European mistletoe was extracted with methanol, purified to partition against diethyl ether and further purified with XAD-7 column chromatography. Pure compounds were separated by Sephadex column chromatography and preparative HPLC. The structures of the novel compounds were established using a combination of several 2D NMR spectroscopic techniques and mass spectrometry.

Results: A new type of natural product derived from the methyl ester of γ-hydroxybutyric acid (GHB) coupled to hydroxybenzoic acids, namely 3-(3′-carbomethoxypropyl) gallic acid and 3-(3′-carbomethoxypropyl)-7→3″-protocatechoyl galloate were characterized from European white-berry mistletoe. Condensation of the 3-hydroxyl of gallic acid with the 4-hydroxyl of GHB significantly reduced the radical scavenging properties of the former compound.

Discussion and conclusion: The characterized compounds define a novel group of natural products that may be of particular interest because it appears that the two new compounds are not closely related to any known natural product.     

北沙参中的新8-O-4′型异木脂素

为了研究北沙参的化学成分，通过大孔树脂吸附柱色谱、 Sephadex LH-20及反相ODS柱色谱进行化合物的分离， 利用多种波谱技术鉴定化合物结构。从醇提物的乙酸乙酯溶部分， 分离得到1个新化合物， 命名为可来灵素D(glehlinoside D，1)， 以及5个已知化合物： 裂异落叶松脂醇(2)、 阿魏酸(3)、 咖啡酸(4)、 香草酸(5)和丁香苷(6)。化合物1为一新的8-O-4′型异木脂素类化合物。     

生川乌瓜蒌不同配伍中乌头类生物碱在大鼠体内的药动学研究

目的考察生川乌瓜蒌不同配伍比例醇提液中乌头类生物碱在大鼠体内的药动学研究,比较瓜蒌对其药动学行为的影响。方法建立灵敏、专属、快速的测定大鼠血浆中6种乌头类生物碱的液相色谱-串联质谱方法。大鼠随机分为3组,分别ig生川乌与瓜蒌1∶0、6∶1、1∶6配伍醇提液,在不同的时间点采血分析,经DAS 2.0软件计算主要药动学参数。结果乌头碱、次乌头碱、新乌头碱的药动学参数tmax和t1/2发生了显著变化,且变化趋势基本一致,在生川乌与瓜蒌6∶1配伍组tmax显著减小(P0.05),生川乌与瓜蒌1∶6配伍组tmax显著增加(P0.01、0.001),且t_(1/2)显著降低(P0.05、0.01),提示生川乌与瓜蒌不同配伍比例影响了3种生物碱的吸收和消除过程。结论生川乌与瓜蒌配伍影响了乌头类生物碱的药动学过程,配伍比例是其配伍禁忌的重要条件。     

Cis- and trans-1,2-diphenylaziridines: induction of xenobiotic-metabolizing enzymes in rat liver and mutagenicity in Salmonella typhimurium

trans-Stilbene imine (trans-1,2-diphenylaziridine) is the nitrogen analog of trans-stilbene oxide, a potent inducer of several microsomal and cytosolic xenobiotic-metabolizing enzymes. Although the acute toxicity of cis- and trans-stilbene imines prevents their application at the usual dose for trans-stilbene oxide (400 mg/kg/day), it is apparent that the imines nevertheless potently induce several xenobiotic-metabolizing enzymes in rat liver. The IP administration of trans-stilbene imine resulted in statistically significant increases in the activities of aminopyrine N-demethylase, microsomal epoxide hydrolase, glutathione transferase (toward 1-chloro-2,4-dinitrobenzene, 1,2-dichloro-4-nitrobenzene and ⁵-androstene-3,17-dione) and UDP-glucuronosyltransferase (toward testoster-one). cis-Stilbene imine was less potent in inducing these activities. Although trans-stilbene imine (total dose = 400 mg/kg) was more potent than trans-stilbene oxide (total dose = 1200 mg/kg) in inducing the activities of glutathione transferase (toward 1-chloro-2,4-dinitrobenzene) and UDP-glucuronosyltransferase (toward testosterone), both compounds belong to the class of substances which are more potent inducers of conjugating (phase II) enzymes.Because of their structural similarity with K-region arene imines which are potent mutagens, cis-stilbene imine and trans-stilbene imine were investigated for mutagenicity (reversion of his ^– strains of Salmonella typhimurium). cis-Stilbene imine and trans-stilbene imine were direct mutagens in the strain TA100. This result, and the finding that acenaphthene 1,2-imine efficiently reverts various strains of Salmonella typhimurium, demonstrates that not only K-region arene imines, but also other aziridines substituted at the two carbons with aromatic moieties, are mutagenic.Abbreviations CSI cis-stilbene imine (cis-2,3-diphenylaziridine) - CSO cis-stilbene oxide - GSH-Px Glutathion peroxidase (activity contributed by a selenium containing enzyme, E. C.1.11.1.9, and certain GST subunits) - GST glutathione transferase (E.C.2.5.1.18) - MC 3-methylcholanthrene - PB phenobarbital - TSI trans-stilbene imine (trans-2,3-diphenylaziridine) - TSO trans-stilbene oxide - UDP-GT uridine-5-diphosphate glucurono-syltransferase (E.C.2.4.1.17) Part of Ph. D. Theses of M. A., P. R., and H. S.     

Identification of Bacteria Isolated from an Oligotrophic Lake with Pesticide Removal Capacities

We studied the growth and capacities for pesticides removal of bacterial strains isolated from the Laguna Grande, an oligotrophic lake at the South of Spain (Archidona, Málaga). Strains were isolated from water samples amended with 10 and 50 g/ml of nine pesticides: organochlorinated insecticides (aldrin and lindane), organophosphorous insecticides (dimetoate, methyl-parathion and methidation), s-triazine herbicides (simazine and atrazine), fungicide (captan) and diflubenzuron (1-(-4-chlorophenyl)-3-(2,6-difluorobenzoyl urea), a chitinase inhibitor. The majority of the strains belonged to the genera Pseudomonas and Aeromonas and only 9% of the total of strains were Gram positive. From all the strains isolated, only 22 showed a wide growth range in all the pesticides tested and 4 of them were chosen for pesticide removal studies. The genetic identification of these strains showed their affiliation to Pseudomonas pseudoalcaligenes, Micrococcus luteus, Bacillus sp. and Exiguobacterium aurantiacum. These last two strains were those that showed the highest pesticide removal capacities and a high bacterial growth.     

Antileishmanial amides and lignans from <Emphasis Type="Italic">Piper cubeba</Emphasis> and <Emphasis Type="Italic">Piper retrofractum</Emphasis>

The n-hexane, ethyl acetate, methanol, and acetone extracts of Piper cubeba Linn. and P. retrofractum Vahl. (Piperaceae) were evaluated in vitro against promastigotes of Leishmania donovani, and all exhibited significant in vitro activity at 100 μg/ml. Two lignans, cubebin and hinokinin, were isolated from the hexane extract of P. cubeba; and one bis-epoxy lignan, (−)-sesamin, and two amides, pellitorine and piplartine, were isolated from the hexane and methanol extracts of P. retrofractum. Cubebin and piplartine showed significant antileishmanial activity in vitro at 100 μM and were further tested in vivo in a hamster model of visceral leishmaniasis. Piplartine showed activity at 30 mg/kg dose. This is the first report of antileishmanial activity of these two plants and their isolated constituents. NIPER Communication No. 395.     

Polymorphism of GSTM1 and GSTT1 genes in bladder cancer: a study from North India

The present study was conducted (1) to examine whether the GSTT1- and GSTM1-null genotypes are risk factors for bladder cancer, and (2) to study possible association of tobacco usage and age strata with genotype of these patients. This case control study was undertaken over a period of 19 months and included 106 bladder cancer patients and 182 controls; both patients and controls originated from northern part of India. The GSTT1 and GSTM1 genotypes were identified by multiplex PCR in peripheral blood DNA samples. Genotype frequencies among patients and controls were assessed and the association of the genotypes with smoking habits and gender of the patients were statistically determined by the ² test. Frequencies of null genotypes in GSTT1 and GSTM1, were 16% (29/182) and 30% (54/182), respectively, in control individuals. The frequencies of GSTT1- and GSTM1-null genotypes in bladder cancer patients were 26% (28/106) and 40% (42/106), respectively. In conclusion, our study demonstrated that the null genotypes of GSTT1 and GSTM1 were substantially at higher risk for bladder carcinoma compared to the normal healthy controls. The GSTT1- and GSTM1-null genotypes did not show significant association with tobacco usage in bladder cancer patients. However, the null genotypes were statistically significant in female relative to male bladder cancer patients.     

Antifungal Activity of a New Triterpenoid Glycoside from Pithecellobium racemosum (M.)

Purpose. In a continuation of our search for novel antifungal compounds from higher plants, the standard extract of the bark of Pithecellobium racemosum was found to have good activity against important AIDS-related opportunistic yeasts. Methods. The extract was subjected to bioguided fractionation using silica gel column chromatography which led to purification of triterpene glycosides. The structures of these compounds were determined by a combination of spectroscopic (IR, NMR, HRMS) and chemical methods. Results. Compound 1 is a new glycoside, 3-O[-L-arabinopyranosyl (1 -2)][-L arabinopyranosyl (1 -6)]2-acetoamido-2-deoxy--D-gluco-pyranosyl oleanolic acid and Compound 2 was identified as the known compound 3-O-[-L-arabinopyranosyl (l-2)]-L-arabinopyranosyl (1-6)] 2-acetamido-2-deoxy--D-glucopyranosyl echinocystic acid. Conclusions. Compound 1 is a new glycoside, 3-O-[-L-arabinopyranosyl (1-2)]-L-arabinopyranosyl (l-6)]-2-acetoamido-2-deoxy--D-glucopyranosyl oleanolic acid and exhibits moderate antifungal activity against T. mentogrophytes, C. albicans and S. cerevisiae with MIC values of 6.25, 12.5 and 12.5 g/ml respectively.