Evaluation of hexane and ethyl acetate extracts of the sponge Jaspis diastra collected from Mauritius Waters on HeLa cells

Objectives Based on previous screening results, the cytotoxic effect of the hexane (JDH) and ethyl acetate extracts (JDE) of the marine sponge Jaspis diastra were evaluated on HeLa cells and the present study aimed at determining their possible mechanism of cell death. Methods Nuclear staining, membrane potential change, flow cytometry analysis of cell cycle distribution and annexin V staining were undertaken to investigate the effects of JDE and JDH. Electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance were used to characterize an isolated bioactive molecule. Key findings JDE displayed an IC50 25 times more significant than the JDH. Flow cytometry analysis revealed JDE induced apoptosis in HeLa cells accompanied by the collapse of mitochondrial membrane potential. Fractionation of JDE resulted in the isolation of the known cytotoxic cyclodepsipeptide, Jaspamide. Conclusions Taking our results together suggest that JDE can be valuable for the development of anticancer drugs, especially for cervical cancer. Further investigations are currently in progress with the aim to determine and isolate other bioactive compounds from this extract.

Effects of Withania somnifera and Tinospora cordifolia Extracts on the Side Population Phenotype of Human Epithelial Cancer Cells: Toward Targeting Multidrug Resistance in Cancer

Recent reports suggest the existence of a subpopulation of stem-like cancer cells, termed as cancer stem cells (CSCs), which bear functional and phenotypic resemblance with the adult, tissue-resident stem cells. Side population (SP) assay based on differential efflux of Hoechst 33342 has been effectively used for the isolation of CSCs. The drug resistance properties of SP cells are typically due to the increased expression of ABC transporters leading to drug efflux. Conventionally used chemotherapeutic drugs may often leads to an enrichment of SP, revealing their inability to target the drug-resistant SP and CSCs. Thus, identification of agents that can reduce the SP phenotype is currently in vogue in cancer therapeutics. Withania somnifera (WS) and Tinospora cordifolia (TC) have been used in Ayurveda for treating various diseases, including cancer. In the current study, we have investigated the effects of ethanolic (ET) extracts of WS and TC on the cancer SP phenotype. Interestingly, we found significant decrease in SP on treatment with TC-ET, but not with WS-ET. The SP-inhibitory TC-ET was further fractionated into petroleum ether (TC-PET), dichloromethane (TC-DCM), and n-butyl alcohol (TC-nBT) fractions using bioactivity-guided fractionation. Our data revealed that TC-PET and TC-DCM, but not TC-nBT, significantly inhibited SP in a dose-dependent manner. Furthermore, flow cytometry-based functional assays revealed that TC-PET and TC-DCM significantly inhibited ABC-B1 and ABC-G2 transporters and sensitized cancer cells toward chemotherapeutic drug-mediated cytotoxicity. Thus, the TC-PET and TC-DCM may harbor phytochemicals with the potential to reverse the drug-resistant phenotype, thus improving the efficacy of cancer chemotherapy.

Spectrophotometric determination of molybdenum with Syzygium jambolanum DC leaf extracts

A new spectrophotometric method for the determination of molybdenum in industrial materials has been developed using the leaf extract of Syzygium jambolanum DC based on the reaction of Mo (VI) at pH 7.0 to produce an orange-yellow complex with an absorption maximum at 426 nm. The molar absorptivity of the complex is 4.27 x 10(4) l mol(-1) cm(-1) and the absorbance, is linear in the range 0.05-0.8 ppm. Sandell sensitivity coefficient was found to be 2.25 x 10(-3) mu g/cm(2). The method is ten times more sensitive than the aqueous thiocyanate system. It has been applied successfully in micronutrient fertilizer, artificial freshwater and sea-water analyses.

Effect of Cattail (Typha domingensis) Extracts, Leachates, and Selected Phenolic Compounds on Rates of Oxygen Production by Salvinia (Salvinia minima)

Salvinia (Salvinia minima Willd.) is a water fern found in Florida waters, usually associated with Lemna and other small free-floating species. Due to its buoyancy and mat-forming abilities, it is spread by moving waters. In 1994, salvinia was reported to be present in 247 water bodies in the state (out of 451 surveyed public waters, Schardt 1997). It is a small, rapidly growing species that can become a nuisance due to its explosive growth rates and its ability to shade underwater life (Oliver 1993). Any efforts toward management of salvinia populations must consider that, in reasonable amounts, its presence is desirable since it plays an important role in the overall ecosystem balance. New management alternatives need to be explored besides the conventional herbicide treatments; for example, it has been shown that the growth of S. molesta can be inhibited by extracts of the tropical weed parthenium (Parthenium hysterophorus) and its purified toxin parthenin (Pande 1994, 1996). We believe that cattail, Typha spp. may be a candidate for control of S. minima infestations. Cattail is an aggressive aquatic plant, and has the ability to expand over areas that weren't previously occupied by other species (Gallardo et al. 1998a and references cited there). In South Florida, T. domingensis is a natural component of the Everglades ecosystem, but in many cases it has become the dominant marsh species, outcompeting other native plants. In Florida public waters, this cattail species is the most dominant emergent species of aquatic plants (Schardt 1997). Several factors enable it to accomplish opportunistic expansion, including size, growth habits, adaptability to changes in the surroundings, and the release of compounds that can prevent the growth and development of other species. We have been concerned in the past with the inhibitory effects of the T. domingensis extracts, and the phenolic compounds mentioned before, towards the growth and propagation of S. minima (Gallardo et al. 1998b). This investigation deals with the impact of cattail materials on the rates of oxygen production of salvinia, as determined through a series of Warburg experiments (Martin et al. 1987, Prindle and Martin 1996).

Effects of extracts of dried seeds of Toloache, Datura innoxia as anaesthesia on the African catfish Clarias gariepinus fingerlings

The effects of crude extract, pure extract, aqueous, fraction of pure and lipid fraction of pure extract of dried seeds of toloache. Datura innoxia as anaesthesia on the African catfish. Clarias gariepinus fingerlings were studied. The fish were exposed to various doses of the extract in aquaria tanks and the time taken for each fish to reach anaesthesia was recorded. The fish were anaesthetized up to 3.00g/l fingerlings reached anaesthesia is significantly (P<0.05) shorter time (1.004 minutes at 0.05gl) in pure unseparated extract than in crude extract (58.50 minutes at 3.00g/l concentrated). The time to reach anaesthesia decreased with an increase in concentration of the seed extract. Out the two fractions the lipid fraction had significantly (P>0.05) better anaesthetic on the fish. The control produced no observable anaesthetic effect on the fish within three hours. This suggests that the anaesthetizing active ingredent resided in the lipid fraction. All fish recovered from anaesthesia, swam and fed actively and no mortality was observed throughout the exposure period and thereafter. It is therefore recommended for use on C. gariepinus fingerlings

Studies of the effects from extracts from two local plants (Tephrosia vogelii and Parkia clappertoniana) on Clarias gariepinus (Tengels)

Studies were carried out using 96hr static toxicity bioassay to determine the effect of lethal concentrations of extracts from two local plants Tephrosia vogelii and Parkia clappertoniana which are known fish poison, on a species of mud fish. Clarias gariepinus Phytochemical analysis of the plant extracts was done and the extract from T. vogelii was found to contain alkaloids, tannins and flavonoids, while the extract from P. clappertoniana was formed to contain alkaloids tannins and saponins. Experimental fish were exposed to test water separately polluted by varying concentrations of extraction of both plant species ranging from 0.50mgl super(-1), 1.50mgl super(-1), 2.50mgl super(-1), 3.0mgl super(-1), 5.00mgl super(-1), 10.00mgl super(-1) in the case of T. vogelii and 5.00mgl super(-1), 7.50mgl super(-1), 10.00mgl super(-1), 15.00mgl super(-1), 20.00mgl super(-1) and 30.00mgl super(-1) in the case of P. clappertaniana. Behavioural hispathological and heamatological examinations were made. Both plant extracts were found to have lethal effects at the higher concentrations, affecting the gills and the central nervous system as well as having a depressive effect on the total count and increasing platelet and white blood cell count. Symptoms of toxicosis observed include, initial inactivation agitated swimming, tumbling movement air gulping, increased opercular beat and period of quiescence/knockdown before death. Marked differences were also observed in the hematological and histopathological analysis of poisoned fish. Lower concentrations of the extracts had sub lethal effects on the fish, which manifested as zigzag movement air gulping increased opercular movement etc. None of these effects were observed in the control experiment

Inhibition of Protease Activity in Muscle Extracts and Surimi from Pacific Whiting, Merluccius productus, and Arrowtooth Flounder, Atheresthes stomias

Muscle extracts of Pacific whiting, Merluccius productus, and arrowtooth flounder, Atheresthes stomias, were assayed for proteolytic activity using azocasein as a substrate. Pacific whiting extracts showed maximum activity at pH 5.0-5.2 and a temperature of 50°C, while arrowtooth flounder extracts had maximum activity at pH 5.5 and 55°C. Three sources of inhibitors (potatoes, egg white, beef plasma protein) were evaluated in vitro for inhibition of protease activity. All three were found to be effective inhibitors in crude muscle extracts. Further studies utilizing these inhibitors in surimi showed that potato was equivalent to both egg white and beef plasma protein in preserving the gel forming characteristics ofheated kamaboko in both species.

Porphyrin detection in denatured cnidarian tissue extracts

Porphyrin metabolic disruption from exposure to xenobiotic contaminants such as heavy metals, dioxins, and aromatic hydrocarbons can elicit overproduction of porphyrins. Measurement of porphyrin levels, when used in conjunction with other diagnostic assays, can help elucidate an organism’s physiological condition and provide evidence for exposure to certain toxicants. A sensitive microplate fluorometric assay has been optimized for detecting total porphyrin levels in detergent solubilized protein extracts from symbiotic, dinoflagellate containing cnidarian tissues. The denaturing buffer used in this modified assay contains a number of potentially interfering components (e.g., sodium dodecyl sulfate (SDS), dithiothreitol (DTT), protease inhibitors, and chlorophyll from the symbiotic zooxanthellae), which required examination and validation. Examination of buffer components were validated for use in this porphyrin assay; while the use of a specific spectrofluorometric filter (excitation 400 ± 15 nm; emission 600 ± 20 nm) minimized chlorophyll interference. The detection limit for this assay is 10 fmol of total porphyrin per μg of total soluble protein and linearity is maintained up to 5000 fmol. The ability to measure total porphyrins in a SDS protein extract now allows a single extract to be used in multiple assays. This is an advantage over classical methods, particularly when tissue samples are limiting, as is often the case with coral due to availability and collection permit restrictions.

Radical Scavenging and Antimicrobial Activities of Methanol Extracts of Two Kinds of Formicary

对两种不同的蚂蚁窝进行甲醇提取,并对提取物进行了急性毒性、抗自由基,酚含量、还原能力和抗菌活性的检测。二苯基三硝基肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基的清除能力的检测表明:两种蚂蚁窝甲醇提取物都具有显著的自由基清除能力,并呈现出剂量依赖性,而且,在高浓度时(5~10mg·mL-1),它们的自由基清除能力甚至显著地高于二丁基羟基甲苯(butylated hydroxyl toluene,BHT)。酚的含量和还原能力分别用福林酚试剂和铁氰化钾还原反应来测定,统计分析显示两种蚂蚁窝的抗氧化能力和其酚含量、还原能力都存在着正相关。两种蚂蚁窝甲醇提取物都具有抗革兰氏阴性菌和革兰氏阳性菌的活性,但它们却对真菌没有作用。

Immnunomodulatory and cytotoxic effects of aqueous leaf extracts of certain plants on common carp, Cyprinus carpio

Effect of aqueous leaf extracts of Catheranthus roseus, Calotropis gigantium and Datura stromoneum on common carp, Cyprinus carpio were investigated. C. carpio were separately fed with 1 and 2% aqueous extracts of these three plant leaves for a period of seven days. In 1% Catheranthus roseus of leaf extract fed group no significant tissue level changes were recorded. One and 2% of other two species fed treated group showed mild to severe necrotic and cellular changes in liver, kidney and spleen. Immunologically, significant rise in antibody titre and respiratory burst activity was recorded for 1% Catheranthus roseusfed group.

Anti-HIV-1 activities of extracts from the medicinal plant Rhus chinensis

Rhus chinensis, a species used in folk medicine by Chinese native people, the anti-HIV-1 activities of the petroleum ether, ethyl acetate, butanol and aqueous extract of Rhus chinensis, named as RC-1, RC-2, RC-3 and RC-4, respectively, was evaluated. The

The effect of Rosmarinus officinalis and Zataria multiflora extracts on the stability of poly unsaturated fatty acids in frozen silver carp minced

The purpose of this study, Evaluation the effect of Rosmarinus officinalis and Thymus vulgaris extracts on the stability of poly unsaturated fatty acids in frozen Silver carp minced. Treatments include: Treatment 1 - Control: frozen meat packaged in conventional Treatment 2: Frozen Silver carp minced+Thyme 300 mg/kg in normal packaging Treatment 3: Frozen Silver carp minced+Rosemary 200 mg/kg in normal packaging Treatment 4: Frozen Silver carp minced+Rosemary compound (100 mg/kg) and Thyme (100 mg/kg) in normal packaging After rapid freezing of samples in the spiral freezer by individual quick freezing method, to maintain the cold temperature (-18) °C were transferred. Sampling and measurements to determine the fatty acid profile of the zero phase beginning in the first month and then every ten days, and 15 days in the second month of the third month after the monthly test. Identifying, defining and measuring the fatty acid profile by gas chromatography was performed. In this study, levels of both saturated and unsaturated fatty acids in three experimental and one control were identified as follows: A: saturated fatty acids: Meristic C14: 0/Palmitic C16: 0/Hepta decaenoic C17: 0/Stearic C18: 0/Arashidic C20: 0/B:Mono unsaturated fatty acids: palmitoleic C16: 1-W7/Oleic C18: 1-W9/Gadoleic C20: 1-W9 C:Poly unsaturated fatty acids: Linoleic C18: 2-W6/α-Linolenic C18: 3-W3 D:High unsaturated fatty acids: Arachidonic C20: 4-W6 Eicosapentaenoic acid C20: 5-EPA/W3 Docosahexaenoic C22: 6-DHA/W3 Results of this study was to determine, Thyme and rosemary extracts containing silver carp minced stored in freezing conditions, Stability of different types of fatty acids, monounsaturated fatty acids, poly-unsaturated fatty acids, omega-3 and omega-6 fatty acids are. So that none of the fatty acids measured were not significant 100% increase or decrease, While changes in the fatty acid oxidation during storage time is minimized. The results obtained from the fatty acid profiles and indicators of their and statistical tests show that treatment with rosemary extract More stable during storage (-18) ° C In comparison with the control and other treatments are shown; And at relatively low compared to other treatments and control samples oleic acid and linoleic acid, palmitic more. According to studies,in Silver carp minced that containing rosemary extract, end of the storage period of six months. Were usable, so even rosemary extract the shelf-life examples to increase more than six months.

Determination of microcystin-LR and its metabolites in snail (Bellamya aeruginosa), shrimp (Macrobrachium nipponensis) and silver carp (Hypophthalmichthys molitrix) from Lake Taihu, China

This paper describes seasonal changes of microcystin-LR (MC-LR) and its glutathione (MC-LR-GSH) and cysteine conjugates (MC-LR-Cys) in three aquatic animals - snail (Bellamya aeruginosa), shrimp (fMacrobrachium nipponensis) and silver carp (Hypophthalmichthys molitrix) collected from Lake Taihu, China. MC-LR, MC-LR-GSH, and MC-LR-Cys were determined by liquid chromatography electrospray ionization mass spectrum (LC-ESI-MS). The mean MC-LR concentrations in the hepatopancreas of snail and shrimp and liver of silver carp were 6.61, 0.24, and 0.027 mu g g(-1) dry weight (DW), respectively: while the average MC-LR-Cys concentrations were 0.50, 0.97, and 5.72 mu g g(-1) DW, respectively. MC-LR-GSH was usually not detectable in these samples. The above results suggest that: (1) in aquatic animals, especially fish, the main excretion form of MC-LR could be MC-LR-Cys, but not MC-LR-GSH, whereas MC-LR-Cys might play an important role in detoxication of MC-LR and (2) that efficiency of MC-LR-Cys formation differs among species. The main detoxication pathway of MC-LR in aquatic animals is suggested as follows: when MC-LR enters into liver/hepatopancreas, it firstly conjugates with polypeptide or protein (including GSH, PP-1 and 2A) containing Cys residues, perhaps also some free cysteine; subsequently, MC-LR-Cys is degraded from these polypeptide or protein; and finally is excreted from animals by the compound of MC-LR-Cys. (C) 2009 Elsevier Ltd. All rights reserved.

Morphological and ultrastructural changes in tobacco BY-2 cells exposed to microcystin-RR

Tobacco BY-2 cells were exposed to microcystin-RR (MC-RR) at two concentrations, 60 mu g mL(-1) and 120 mu g mL(-1), to study the changes in morphology and ultrastructure of cells as a result of the exposure. Exposure to the lower concentration for 5 d led to typical apoptotic morphological changes including condensation of nuclear chromatin, creation of a characteristic 'half moon' structure, and cytoplasm shrinkage and decreased cell volume, as revealed through light microscopy. fluorescence microscopy, and transmission electron microscopy, respectively. Exposure to the higher concentration, on the other hand, led to morphological and ultrastructural changes typical of necrosis, such as rupture of the plasma membrane and the nuclear membrane and a marked swelling of cells. The presence of many vacuoles containing unusual deposits points to the involvement of vacuoles in detoxifying MC-RR. Results of the present study indicate that exposure of tobacco BY-2 cells to MC-RR at a lower concentration (60 mu g mL(-1)) results in apoptosis and that to a higher concentration (120 mu g mL(-1)), in necrosis. (C) 2009 Elsevier Ltd. All rights reserved.