Crude Dichloromethane Research Articles

Gastroprotective, ulcer healing and antioxidant activity of protoberberine alkaloid isolated from partially purified fraction of crude dichloromethane stem extract of coscinium fenestratum

Protoberberine alkaloids such as berberine, jatrarrhizine, and palmatine have been reported to be a major constituent in C. fenestratum. Formulations of these are alkaloids are widely used in traditional systems of medicine including, Ayurveda and Traditional Chinese Medicine and has demonstrated wide range of pharmacological activities including; antihypertensive, anti‐inflammatory, antioxidant, antidepressant, anticancer, antidiarrhoeal, antidiabetic cholagouge, hepatoprotective and anti‐microbial. Our previous study showed that crude and partially purified fraction (PPF) obtained from dichloromethane (DCM) stem extract of C.fenestratum has a very significant antiulcer activity. Therefore in this study we evaluated the ulcer healing and antioxidant effects of PPF of DCM stem extract Coscinium fenestratum. The ability of the PPF to heal gastric mucosa against injuries caused by necrotizing agent's 0.3M HCl/60% EtOH, indomethacin and stress were investigated. Determination of DPPH, hydroxyl, nitric oxide and superoxide radical‐scavenging activity (antioxidant), was performed using simple biochemical reactions and LC‐MS profiling of the bioactive compound was also determined. PPF demonstrated a significant (p< 0.001) ulcer healing effect, in HCl/EtOH‐induced gastric ulcer, it reduced ulcerative lesion index by 76.2%, stress 93.5%, indomethacin was 74.60%, acetic acid 75% compared to negative control group. DPPH radical scavenging activity showed IC50 value of 0.194mg/ml and superoxide radical scavenging activity IC50 value of 1.54 mg/ml. LC‐MS profiling indicated the presence of Palmitine.

In vitro antiplasmodial and cytotoxic properties of some medicinal plants from western Burkina Faso.

BackgroundResistance of malaria parasites to existing drugs complicates treatment, but an antimalarial vaccine that could protect against this disease is not yet available. It is therefore necessary to find new effective and affordable medicines. Medicinal plants could be a potential source of antimalarial agents. Some medicinal plants from Burkina Faso were evaluated for their antiplasmodial and cytotoxic properties in vitro.MethodsCrude dichloromethane, methanol, water-methanol, aqueous and alkaloids extracts were prepared for 12 parts of 10 plants. Chloroquine-resistant malaria strain K1 was used for the in vitro sensibility assay. The Plasmodium lactacte dehydrogenase technique was used to determine the 50% inhibitory concentration of parasites activity (IC50). The cytotoxic effects were determined with HepG2 cells, using the tetrazolium-based colorimetric technique, and the selectivity index (SI) was calculated.ResultsSixty crude extracts were prepared. Seven extracts from Terminalia avicenoides showed IC50 < 5 µg/mL. The IC50 of dichloromethane, methanol, aqueous and alkaloids extracts ranged between 1.6 µg/mL and 4.5 µg/mL. Three crude extracts from Combretum collinum and three from Ficus capraefolia had an IC50 ranging between 0.2 µg/mL and 2.5 µg/mL. Crude extracts from these three plants had no cytotoxic effect, with SI > 1. The other plants have mostly moderate or no antimalarial effects. Some extracts from Cordia myxa, Ficus capraefolia and Opilia celtidifolia showed cytotoxicity, with an SI ranging between 0.4 and 0.9.ConclusionOur study showed a good antiplasmodial in vitro activity of Terminalia avicenoides, Combretum collinum and Ficus capraefolia. These three plants may contain antiplasmodial molecules that could be isolated by bio-guided phytochemical studies.

The In-vivo Study of Anti-asthmatic Effect of Partially Purified Extracts From The Leaves of Labisia Pumila

Labisia pumila popularly known as Kacip Fatimah is a sub herbaceous plant which belonging to the Myrsinaceae family. It has been used in folk medicine to treat dysenteries and inflammatory disorders, such as rheumatism. Our previous study showed that crude dichloromethane extract exhibited a very strong anti-asthmatic and mast-cell stabilizing activity. In this present study, partially purified fractions A-E, of dichloromethane (DELP) crude extract obtained by column chromatography techniques were studied for their anti-asthmatic effect. 1% histamine, serotonin and bradykinin (0.1ml/rat) was dissolved with 0.9% (w/v) saline and subcutaneously injected to induce edema formation. 2mg of arachidonic acid was dissolved in 20μl of acetone and topically applied to mouse ear to induce ear edema. Also, 1% histamine and acetylcholine was dissolved in saline and was used to induce bronchoconstriction in guinea pigs. Oral treatment of partially purified fractions A-E (100mg/kg), and indomethacin (10mg/kg) inhibited significantly (p<0.001) the formation of the histamine, serotonin and bradykinin-induced paw edema measured in the fifth hour. The highest inhibition was observed in fraction E which was 5.07% 5.63% for histamine, 5.65% for serotonin, 9.59% for bradykinin and 43% for arachidonic acid ear edema. Fraction A showed 55% and 48.17% respectively for inhibition of histamine and acetylcholine-induced bronchoconstriction after 24 hrs treatment and 68% and 51.21% protection after 14 days treatment. Phytochemical screening revealed the presence of flavonoids, steroids, saponins, alkaloids and tannins. The present study confirmed that the extracts exhibited anti-asthmatic activity by its antiinflammatory and bronchodilatory action.

Antioxidant activities and triterpenoids isolated from Alternanthera brasiliana (L.) Kuntze leaves

The species Alternanthera brasiliana (L.) Kuntze (Amaranthaceae) is a perennial herbaceous plant widely distributed in Brazil's eastern beaches and certain Amazonian regions. Crude ethanolic (70%) extract, fractions (dichloromethane, ethyl acetate and n-butanolic) and the mixture isolated were investigated regarding their content of total phenolic compounds by Folin-Ciocalteau assay. Total phenolic contents varied from 29.0 to 56.6 mg/g (dry fraction). The scavenging capacity of the extract, fractions and mixture isolated was characterised by the 1,1-diphenyl-2-picrylhydrazyl method. Only ethyl acetate fraction exhibited a modest scavenging capacity (SC50 = 163.00 μg/mL and SC% = 64.29% at 250 μg/mL) in a dose-dependent manner. Crude extract, dichloromethane and butanolic fractions presented a very weak scavenging capacity (SC% ≤ 40% at 250 μg/mL). A correlation between the antioxidant activity and the total phenolic contents could not be observed. The fractionation and purification of dichloromethane fraction evidenced the presence of a mixture of β-sitosterol, stigmasterol and spinasterol, which was identified by NMR methods and compared with literature data. Results of this study demonstrated for the first time the antioxidant activity of A. brasiliana.

Studies of the in vitro cytotoxic, antioxidant, lipase inhibitory and antimicrobial activities of selected Thai medicinal plants

BackgroundTraditional folk medicinal plants have recently become popular and are widely used for primary health care. Since Thailand has a great diversity of indigenous (medicinal) plant species, this research investigated 52 traditionally used species of Thai medicinal plants for their in vitro cytotoxic, antioxidant, lipase inhibitory and antimicrobial activities.MethodsThe 55 dried samples, derived from the medicinally used parts of the 52 plant species were sequentially extracted by hexane, dichloromethane, ethanol and water. These 220 extracts were then screened for in vitro (i) cytotoxicity against four cell lines, derived from human lung (A549), breast (MDA-MB-231), cervical (KB3-1) and colon (SW480) cancers, using the MTT cytotoxicity assay; (ii) antioxidant activity, analyzed by measuring the scavenging activity of DPPH radicals; (iii) lipase inhibitory activity, determined from the hydrolytic reaction of p-nitrophenyllaurate with pancreatic lipase; and (iv) antimicrobial activity against three Gram-positive and two Gram-negative bacteria species plus one strain of yeast using the disc-diffusion method and determination of the minimum inhibitory concentration by the broth micro-dilution assay.ResultsThe crude dichloromethane and/or ethanol extracts from four plant species showed an effective in vitro cytotoxic activity against the human cancer cell lines that was broadly similar to that of the specific chemotherapy drugs (etoposide, doxorubicin, vinblastine and oxaliplatin). In particular, this is the first report of the strong in vitro cytotoxic activity of Bauhinia strychnifolia vines. The tested tissue parts of only six plant species (Allium sativum, Cocoloba uvifera, Dolichandrone spathacea, Lumnitzera littorea, Sonneratia alba and Sonneratia caseolaris) showed promising potential antioxidant activity, whereas lipase inhibitory activity was only found in the ethanol extract from Coscinum fenestratum and this was weak at 17-fold lower than Orlistat, a known lipase inhibitor. The highest antimicrobial activity was observed in the extracts from S. alba and S. caseolaris against Pseudomonas aeruginosa and Candida albicans, respectively.ConclusionThe Thai medicinal plant B. strychnifolia is first reported to exert strong in vitro cytotoxic activities against human cancer cell lines and warrants further enrichment and characterization. The broad spectrum of the biological activities from the studied plant extracts can be applied as the guideline for the selection of Thai medicinal plant species for further pharmacological and phytochemical investigations.

HPLC Analysis of Polyphenolic Compounds and Antioxidant Activity in Nasturtium officinale

The antioxidant activities of crude extract, dichloromethane, ethyl acetate, and butanolic fractions of Nasturtium officinale were investigated using the 1,1,-diphenyl-2-picryl-hydrazyl and thiobarbitaric acid reactive species methods. Total phenolics and flavonoids were determined. Rutin, chlorogenic, and caffeic acids were quantified in the crude extract by high performance liquid chromatography-photodiode array detector. The antioxidant and radical scavenging activity of the crude extract and fractions were found in the following decreasing order: butanolic fraction > ethyl acetate fraction > dichloromethane fraction > crude extract. The extent of antioxidant activities was in accordance with the amounts of phenolics and flavonoids present in the extract and fractions. Ethyl acetate and butanolic fractions caused a sharp fall in thiobarbitaric acid reactive species production, starting at the concentration of 1 μg/mL for both fractions with IC50 6.92 ± 0.19 and 12.07 ± 0.45 μg/mL, respectively. N. officinale might be a valuable antioxidant natural source and seemed to be applicable in both healthy medicine and the food industry.

Antibacterial and antifungal activities of crude plant extracts from Colombian biodiversity

On a global scale, people have used plants to treat diseases and infections, and this has raised interest on the plant biodiversity potencial in the search of antimicrobial principles. In this work, 75 crude n-hexanes, dichloromethane and methanol extracts from the aerial parts of 25 plants belonging to four botanical families (Asteraceae, Euphorbiaceae, Rubiaceae and Solanaceae), collected at the Natural Regional Park Ucumari (Risaralda, Colombia), were evaluated for their antibacterial and antifungal activities by the agar well diffusion method. The antibacterial activities were assayed against two Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis, and three Gram-negative ones named, Klebsiella pneumoniae, Escherichia coli and Pseudomonas aeruginosa. In addition, the same plant extracts were tested against the yeast Candida albicans and the fungi Aspergillus fumigatus and Fusarium solani. Overall, the plant extracts examined displayed better bactericide rather than fungicide activities. In general, the best antibacterial activity was showed by the plant extracts from the Rubiaceae family, followed in order by the extracts from the Euphorbiaceae and Solanaceae ones. It is important to emphasize the great activity displayed by the methanol extract of Alchornea coelophylla (Euphorbiaceae) that inhibited four out of five bacteria tested (B. Subtilis, P. aeruginosa, S. aureus and E. coli). Furthermore, the best Minimal Inhibitory Concentration for the extracts with antifungal activities were displayed by the dichloromethane extracts from Acalypha diversifolia and Euphorbia sp (Euphorbiaceae). The most susceptible fungus evaluated was F. Solani since 60% and 20% of the dichloromethane and methanol extracts evaluated inhibited the growth of this phytopathogenic fungus. The antimicrobial activity of the different plant extracts examined in this work could be related to the secondary metabolites contents and their interaction and susceptibility of pathogenic microorganism evaluated.

The cytotoxic effect of endemic Centaurea fenzlii Reichardt on colon cancer cell lines

Being the largest genus of the Compositae family with over 169 species and 199 taxons, Centaurea L. has been used in folklore for the treatment of wide range of diseases in Turkey. The antitumor activities of the Centaurea species in different types of cancer have been identified through several bioactivity studies. However, the in vitro anticancer activity of endemic Centaurea fenzlii Reichardt. has never been evaluated and published yet. In this study, the cytotoxic effect of this plant was tested in vitro against Caco-2 (human epithelial colorectal adenocarcinoma) cells by using MTT, and ATP assays. For the positive control, the cells were treated with the chemotherapeutic drug 5-fluorouracil which has been widely used in the treatment of a range of cancers. The aerial parts of the plants were collected during the flowering period and the extracts were obtained with n-hexane, dichloromethane and methanol, respectively. The extracts were applied to Caco-2 cell lines at increasing doses. The IC50 values of samples were determined and their direct cytotoxic effects were measured. The extracts displayed noteworthy cell growth and proliferation inhibitory activity. Moreover, the cytotoxic effect of crude dichloromethane extract on Caco-2 cell lines was the highest compared to other extracts. This cytotoxicity screen has provided important preliminary data promoting the selection of the plant species and their different extracts with potential antitumor properties for a therapeutic option in colon cancer.

Chemical constituents and free radical scavenging activity of corn pollen collected from Apis mellifera hives compared to floral corn pollen at Nan, Thailand

BackgroundBee pollen is composed of floral pollen mixed with nectar and bee secretion that is collected by foraging honey (Apis sp.) and stingless bees. It is rich in nutrients, such as sugars, proteins, lipids, vitamins and flavonoids, and has been ascribed antiproliferative, anti-allergenic, anti-angiogenic and free radical scavenging activities. This research aimed at a preliminary investigation of the chemical constituents and free radical scavenging activity in A. mellifera bee pollen.MethodsBee pollen was directly collected from A. mellifera colonies in Nan province, Thailand, in June, 2010, whilst floral corn (Zea mays L.) pollen was collected from the nearby corn fields. The pollen was then sequentially extracted with methanol, dichloromethane (DCM) and hexane, and each crude extract was tested for free radical scavenging activity using the DPPH assay, evaluating the percentage scavenging activity and the effective concentration at 50% (EC50). The most active crude fraction from the bee pollen was then further enriched for bioactive components by silica gel 60 quick and adsorption or Sephadex LH-20 size exclusion chromatography. The purity of all fractions in each step was observed by thin layer chromatography and the bioactivity assessed by the DPPH assay. The chemical structures of the most active fractions were analyzed by nuclear magnetic resonance.ResultsThe crude DCM extract of both the bee corn pollen and floral corn pollen provided the highest active free radical scavenging activity of the three solvent extracts, but it was significantly (over 28-fold) higher in the bee corn pollen (EC50 = 7.42 ± 0.12 μg/ml), than the floral corn pollen (EC50 = 212 ± 13.6% μg/ml). After fractionation to homogeneity, the phenolic hydroquinone and the flavone 7-O-R-apigenin were found as the minor and major bioactive compounds, respectively. Bee corn pollen contained a reasonably diverse array of nutritional components, including biotin (56.7 μg/100 g), invert sugar (19.9 g/100 g), vitamin A and β carotene (1.53 mg/100 g).ConclusionsBee pollen derived from corn (Z. mays), a non-toxic or edible plant, provided a better free radical scavenging activity than floral corn pollen.

In vitro antiproliferative/cytotoxic activity on cancer cell lines of a cardanol and a cardol enriched from Thai Apis mellifera propolis

BackgroundPropolis is a complex resinous honeybee product. It is reported to display diverse bioactivities, such as antimicrobial, anti-inflammatory and anti-tumor properties, which are mainly due to phenolic compounds, and especially flavonoids. The diversity of bioactive compounds depends on the geography and climate, since these factors affect the floral diversity. Here, Apis mellifera propolis from Nan province, Thailand, was evaluated for potential anti-cancer activity.MethodsPropolis was sequentially extracted with methanol, dichloromethane and hexane and the cytotoxic activity of each crude extract was assayed for antiproliferative/cytotoxic activity in vitro against five human cell lines derived from duet carcinoma (BT474), undifferentiated lung (Chaco), liver hepatoblastoma (Hep-G2), gastric carcinoma (KATO-III) and colon adenocarcinoma (SW620) cancers. The human foreskin fibroblast cell line (Hs27) was used as a non-transformed control. Those crude extracts that displayed antiproliferative/cytotoxic activity were then further fractionated by column chromatography using TLC-pattern and MTT-cytotoxicity bioassay guided selection of the fractions. The chemical structure of each enriched bioactive compound was analyzed by nuclear magnetic resonance and mass spectroscopy.ResultsThe crude hexane and dichloromethane extracts of propolis displayed antiproliferative/cytotoxic activities with IC50 values across the five cancer cell lines ranging from 41.3 to 52.4 μg/ml and from 43.8 to 53.5 μg/ml, respectively. Two main bioactive components were isolated, one cardanol and one cardol, with broadly similar in vitro antiproliferation/cytotoxicity IC50 values across the five cancer cell lines and the control Hs27 cell line, ranging from 10.8 to 29.3 μg/ml for the cardanol and < 3.13 to 5.97 μg/ml (6.82 - 13.0 μM) for the cardol. Moreover, both compounds induced cytotoxicity and cell death without DNA fragmentation in the cancer cells, but only an antiproliferation response in the control Hs27 cells However, these two compounds did not account for the net antiproliferation/cytotoxic activity of the crude extracts suggesting the existence of other potent compounds or synergistic interactions in the propolis extracts.ConclusionThis is the first report that Thai A. mellifera propolis contains at least two potentially new compounds (a cardanol and a cardol) with potential anti-cancer bioactivity. Both could be alternative antiproliferative agents for future development as anti-cancer drugs.

BRINE SHRIMP LETHALITY BIOASSAY OF THE AERIAL PARTS OF MARISCUSSUMATRENSIS

Mariscus sumatrensis of cyperaceae family is one of the rare plat species of Bangladesh traditionally used by the indigenous people for the treatment of various diseases has been investigated for its cytotoxic property by Brine shrimp assay. Crude methanol extract showed lowest LC50 value than crude petroleum ether and crude dichloromethane extract during the assay

In vitro toxicity, antiplatelet and acetylcholinesterase inhibition of Buddleja thyrsoides Lam. leaves

Alzheimer's disease (AD) is a neurodegenerative disorder resulting in impaired memory and behaviour of remarkable socio-economic impact. A decrease in cholinergic activity is a key event in the biochemical of AD. Buddleja thyrsoides is a plant widely distributed in Southern parts of South America. In Brazilian traditional medicine, the infusion of its leaves and flowers is used for the treatment of bronchitis and cough. Crude ethanolic (70%) extract and fractions (dichloromethane, ethyl acetate and n-butanolic) were investigated regarding their toxicities in vitro and antiplatelet action. The enzyme acetylcholinesterase inhibition was evaluated to study the crude extract. The crude extract and fractions were evaluated by means of Brine Shrimp Lethality test and they showed low activities with LC50 values 1698, 2818, 2187 and 3672 µg mL−1 for dichloromethane, ethyl acetate, n-butanolic fractions and crude extract, respectively. Buddleja thyrsoides presented great antiplatelet action. The IC50 values obtained for crude extract and dichloromethane, ethyl acetate and n-butanolic fractions were 361.29, 354.23, 368.75 and 344.30, respectively, while the IC50 for the standard AAS was 257.01 µg mL−1. The crude extract showed an inhibition of 22.8% of the acetylcholinesterase enzyme in 24 h.

The antioxidant properties, cytotoxicity and monoamine oxidase inhibition abilities of the crude dichloromethane extract of Tarchonanthus camphoratus L. leaves

Tarchonanthus camphoratus (camphor bush) has been widely used for numerous medicinal purposes. The aim of the present study was to evaluate the antioxidant properties, cytotoxicity and monoamine oxidase inhibition activities of the crude dichloromethane leaf extract of T. camphoratus . The antioxidant activities were assessed using the thiobarbituric acid-reactive substances (TBARS) assay and the nitroblue tetrazolium (NBT) assay. The cytotoxicity assays were performed according to the microculture MTT method. From the MTT assay, it was determined that at a concentration of 10 mg/ml of crude extract, 95% of the neuroblastoma cells were killed. Almost 99% of the cells were viable at a concentration of 0.08 mg/ml extract. The extract also showed the ability to inhibit monoamine oxidase (MAO-A and MAO-B) with the corresponding inhibitory concentration, IC 50 values of 1.371 and 0.2737 mg/ml, respectively. The antioxidant activity and cytotoxic effect of the extracts increased with increase in concentration. This study suggests that the dichloromethane leave extract of T. camphoratus can potentially be used as a readily accessible source of natural antioxidants.

Antiproliferative activity, isolation and identification of active compound from Gaylussacia brasiliensis

Gaylussacia brasiliensis (Spreng.) Meissn., Ericaceae, is used in folk medicine for treatment of several inflammatory processes and as healing agent. The scope of this work was to evaluate the in vitro antiproliferative activity of crude dichloromethane extract (CHD) and to identify the compound(s) responsible for this activity. CHD was evaluated and showed a concentration dependent inhibition on all cells lines. Therefore CHD was submitted to several classical columns chromatography providing the most active fraction (FC), inhibiting all cells line at 25 µg/mL. FC was further fractionated affording isolated compound 2β, 3β-dihydroxy-urs-12-ene-28-oic acid , identified on basis of 2D-NMR experiments and showed concentration-dependent activity and selectivity for kidney and breast cell lines.

Isolation of immunomodulatory triterpene acids from a standardized rose hip powder (Rosa canina L.)

A previously published systematic review and a metaanalysis have concluded that the consumption of standardized rose hip powder (Rosa canina L.) can reduce pain in osteoarthritis patients. Synovial inflammation has been suggested to play an important role in the pathogenesis of osteoarthritis and mainly to involve infiltration of the synovial membrane by macrophages. Therefore, the immunomodulatory effect of standardized rose hip powder of Rosa canina L. was investigated and active principles isolated using the Mono Mac 6 cell line as a model for human macrophages. Treatment of Mono Mac 6 cells with the residue of a crude dichloromethane extract of rose hip powder significantly and concentration dependently inhibited the lipopolysaccharide induced interleukin-6 release. Through bioassay-guided fractionation the immunomodulatory effect of the dichloromethane extract was correlated to a mixture of three triterpene acids; oleanolic acid, betulinic acid and ursolic acid (IC(50) 21 ± 6 µm). Further studies revealed that only oleanolic acid and ursolic acid, but not betulinic acid, could inhibit the lipopolysaccharide induced interleukin-6 release from Mono Mac 6 cells when tested separately. Combination of either oleanolic acid or ursolic acid with betulinic acid enhanced the immunomodulatory effect of the two triterpene acids.

Antitrypanosomal Activity of a Semi-Purified Subfraction Rich in Labdane Sesquiterpenes, Obtained from Flowers of Anthemis Tinctoria, Against Trypanosoma Cruzi

In Brazil and several other Latin American countries, Chagas' disease still constitutes a serious medical and social problem, and there is a need to develop new, more-potent drugs with fewer side effects to effectively treat this disease. We investigated the antitrypanosomal effect of a crude extract, fractions, and a semi-purified subfraction rich in a mix- ture of isomeric labdane sesquiterpenes, obtained from flowers of Anthemis tinctoria, against Trypanosoma cruzi. In epimastigote forms, the aqueous crude extract, dichloromethane fraction, and semi-purified subfraction showed a dose-dependent inhibitory activity, with IC50 of 2.3 μg/ml, 1.8 μg/ml, and 0.2 μg/ml, respectively. In the interaction in- dex, the semi-purified subfraction showed a reduction in both the percentage of infected LLCMK2 cells and the mean number of amastigotes per infected cell. The cytotoxicity evaluation demonstrated that the cytotoxic concentrations of the semi-purified subfraction were higher for LLCMK2 cells than for the protozoans, with a selectivity index of 35.0. Epimastigote forms treated with the semi-purified subfraction showed ultrastructural and morphological alterations such as rounding of the cells and bleb formation in the flagellum and cytoplasmic membrane. These results show that the flowers from A. tinctoria may be a source of new drugs with antiprotozoal activity. However, additional in vitro and in vivo studies are needed to validate the use of A. tinctoria in the treatment of Chagas’ disease.

Antioxidant and antitopoisomerase activities in plant extracts of some Colombian flora from La Marcada Natural Regional Park

Many plants have been used to treat some diseases and infections since time immemorial, and this potential has been exploited by the pharmaceutical industry in the search of new analgesic, anticarcinogenic and antimicrobial agents, among other active agents. In order to contribute with bioprospection studies on the Colombian flora, 35 extracts from 13 plant species belonging to seven families (Apocynaceae, Cactaceae, Costaceae, Eremolepidaceae, Passifloraceae, Solanaceae and Urticaceae) were collected from La Marcada Natural Regional Park (LMNRP), Colombia. Dichloromethane, n-hexane and aqueous-methanol crude extracts were prepared and evaluated for their activity against Saccharomyces cerevisiae RS322N, R52Y and RS321 strains in the yeast mutant assay and their antioxidant capacity through the DPPH test. The dichloromethane extract from Myriocarpa stipitata (Urticaceae) showed moderate inhibitory activity against the three S. cerevisiae strains tested. The capacity of the dichloromethane extract from M. stipitata to inhibit the enzyme topoisomerase I and to cause DNA damage was inferred from these results. In the DPPH assay, the n-hexane crude extract from Costus sp. (Costaceae) showed good antioxidant activity (48%); in addition, the crude dichloromethane and aqueous-methanol extracts from Rhipsalis micrantha (Cactaceae) showed moderate antioxidant activity with percentage of 29 and 21%, respectively.

Antiedematogenic activity and phytochemical composition of preparations from Echinodorus grandiflorus leaves

The leaves of Echinodorus grandiflorus (Alismataceae) are traditionally used in Brazil to treat inflammatory conditions. The aim of the present study was to evaluate the antidematogenic activity of crude aqueous, dichloromethane and hydroethanolic extracts from E. grandiflorus leaves using the carrageenan-induced paw edema model in mice, along with of fractions enriched in diterpenes, flavonoids and hydroxycinnamoyltartaric acids (HCTA). Significant inhibitions of paw edema were elicited by the 50% and 70% EtOH extracts (1000 mg/kg, p.o.), as well as by the fractions enriched in diterpenes (70-420 mg/kg, p.o.) and flavonoids (7.2-36 mg/kg, p.o.). Isovitexin, isoorientin, trans-aconitic and chicoric acids were identified in all extracts by HPLC analysis. Trans-aconitic acid itself exhibited significant antiedematogenic effect (270 mg/kg, p.o.). The biological activity correlated positively with the contents of flavonoids and diterpenes, but negatively with HCTA concentrations, demonstrating the participation of the two classes of compounds in the antiedematogenic activity of E. grandiflorus.

Anti-Human Papillomavirus (HPV) 16 E6 Activity of Ling Zhi or Reishi Medicinal Mushroom, Ganoderma lucidum (W. Curt.: Fr.) P. Karst. (Aphyllophoromycetideae) Extracts

Cancer of the cervix ranks second in the demographics of cancers affecting women worldwide. Human papillomavirus (HPV) type 16 is one of the major causative agents for cervical cancer. The E6 region of the HPV 16 genome encodes for an oncoprotein responsible for the pathogenesis of the disease. The E6 oncoprotein is abundant in cervical cancer and cervical cancer-derived cell lines. Mushrooms of the genus Ganoderma have long been used traditionally as an immunomodulating and antitumor agent, with recent studies validating its use in chemotherapeutics. Due to the importance of the E6 oncoprotein in the carcinogenesis of cervical cancer, this study was undertaken to screen the crude dichloromethane, ethanol, water, and polysaccharide extracts of Ganoderma lucidum for the ability to suppress the expression of HPV 16 E6 and to evaluate secondary metabolite classes present in the active crude extract. CaSki cells were treated with the crude extracts and were then subjected to immunocytochemistry protocols. All crude extracts of G. lucidum presented HPV 16 E6 suppression, with the dichloromethane extract being the most active compared to the other crude extracts. The crude dichloromethane extract was then subjected to thin-layer chromatography to evaluate the secondary metabolite classes present. The present study shows that the crude dichloromethane extract of G. lucidum possesses flavonoids, terpenoids, phenolics, and alkaloids. Further isolation work on the dichloromethane extract of G. lucidum could direct us to lead compounds with promising anti-HPV 16 E6 activity.

Alpha glucosidase inhibition by stem extract of Tinospora cordifolia

Inhibitors of alpha glucosidase have potential use in the treatment of diabetes mellitus. The stem extract of Tinospora cordifolia was evaluated for inhibition of the enzyme. The extract was also found to inhibit the salivary and pancreatic amylase and therefore can effectively reduce an increase in postprandial glucose level. The crude ethyl acetate, dichloromethane (DCM), chloroform and hexane extracts of Tinospora cordifolia were studied. 15 mg of the DCM extract was most effective in that showed 100 % inhibition of the alpha glucosidase whereas salivary amylase was inhibited to the extent of 75 % and pancreatic amylase to 83 %. On giving a maltose load of 2mg / g along with 0.3 mg / g body weight of the DCM Tinospora stem extract a decrease was revealed in the hyperglycemic shoot up in normal and diabetic animals by 50 and 58 % respectively as compared to the controls. The extract was found to inhibit alpha glucosidase in a non-competitive manner.