Costunosides A-C: cytotoxic sesquiterpene lactones from the rhizomes of Aucklandia costus Falc.

Three new eudesmane type rare sesquiterpene lactone galactosides, costunosides A-C (1-3) were isolated from the rhizomes of Aucklandia costus along with ten known compounds (4-13). Costunosides A-C (1-3) are the first example of naturally eudesmane glycosides containing a ß-galactopyranoside moiety. The structure and relative configurations of these compounds were established by comprehensive analysis of MS and, in particular 1D/2D NMR spectroscopic data. The isolated compounds were tested against a panel of human cancer cell lines, where compounds 3, 6 and 7 have shown promising cytotoxic activity against PC-3, HCT-116 and A549 cell lines with IC50 values in the range of 3.4 µM to 9.3 µM, respectively. Costunosides A-C (1-3) were also screened for inhibition assay of acetyl-cholinesterase (AChE), and butyrylcholinesterase (BChE) and found inactive at a concentration of 10 µM.

A novel carissic acid from fruits of Carissa carandas induced apoptotic cell death in A-549 cells following the activation of caspases.

This study was designed to identify cytotoxic compounds from Carissa carandas extract. The cytotoxic activity of extract and fractions were assessed against eight cancer cell lines. The chloroform fraction obtained from methanolic extract exhibited significant activity against MCF-7, HT-29, A-549 with IC50 values of 3.98 µg/mL (MCF-7), 1.28µg/mL (HT-29) and 1.48 µg/mL (A-549) respectively. Further investigation led to the isolation of novel compound carissic acid (CA), which was confirmed by detailed spectroscopy studies. CA exhibited notable activity with IC50 values of 3.47 µM for A-549, 2.65 µM for HT-29 and 13.58 ± 0.59 µM for MCF-7 cells. CAcaused chromatin condensation with decrease of mitochondrial membrane potential and also confirmed cell death via Reactive Oxygen Species (ROS) generation and significantly decreased the colony formation in dose-dependent manner. The overall findings suggested that CA demonstrates cytotoxic effect by inhibiting cell proliferation and promoting apoptosis in lung (A-549) carcinoma cell line.

Seroprevalence of human cystic echinococcosis in South Kashmir, India.

Cystic echinococcosis is a zoonotic disease that causes economic losses and public health problems throughout the globe. Present study was undertaken to estimate the seroprevalence of cystic echinococcosis in humans of South Kashmir and to determine the risk factors associated with this disease. The present study was carried out from April 2017 to March 2018, during which 458 blood samples (from 222 males and 236 females) were collected from selected subjects from 12 villages of four districts along with demographic characteristics. Samples were analyzed by an Enzyme-linked Immunosorbent Assay (ELISA) (commercially prepared kit) for detection of immunoglobulin IgG against cystic echinococcosis. Out of 458 samples, 20(4.36%) samples were found positive. Results implied that the seroprevalence of cystic echinococcosis in South Kashmir had significant relationship with age, gender and occupation (P < .05). Seroprevalence for cystic echinococcosis was significantly higher among males, children, and illiterate persons.

Evaluation of anthelmintic antimicrobial and antioxidant activity of Chenopodium album.

The objective of this study was to evaluate the anthelmintic, antimicrobial and antioxidant activities of Chenopodium album against gastrointestinal nematodes of sheep and some pathogenic microbes. A worm motility inhibition assay was used for in vitro study, and a faecal egg count reduction assay was used for an in vivo study. Various concentrations ranging from 100 to 500 µg/ml of the extract were subjected to antimicrobial screening by disc diffusion method against four selected bacterial (Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas multocida and Escherichia coli) and two fungal (Aspergillus flavus and Candida albicans) strains in order to estimate the medicinal potential of the herb. DPPH (1,1-diphenyl-2-picrylhydrazyl), riboflavin photo-oxidation, deoxyribose, lipid peroxidation assays were used for antioxidant activity. The extracts exhibited dose- and time-dependent anthelmintic effects on the Haemonchus contortus as compared to levamisole. The extract showed maximum inhibitory effect against S. aureus (28 ± 0.14 mm), while as mild inhibitory effect was observed against E. coli among the selected microbial strains. The effect produced by the different extract concentrations was comparable with the standard antibacterial agent streptomycin sulphate and antifungal agent nystatin, which were used as effective positive control in the study. The antioxidant activity showed that the extracts exhibited scavenging effect in concentration-dependent manner on superoxide anion radicals and hydroxyl radicals leading to the conclusion that the plant has broad spectrum anthelmintic, antimicrobial and antioxidant activities and could be a potential alternative for treating various diseases.

Assessment of species richness in Lake Dal, Kashmir, based on classical approach, physiological approach and rDNA ITS sequences from isolates.

As a first description to document the species richness in Dal Lake, a freshwater lake ecosystem in Kashmir valley, an extensive network of sixteen sampling stations with distinguishing features was sampled seasonally for two years. The identification process yielded fifty-one species probably first and new records for this area to date. The taxonomic groups observed were those with species from Ascomycetes (inclusive of yeasts), Basidiomycetes, Blastocladiomycetes, Zygomycetes, and Peronosporomycetes. Each phylum was represented by a single Order, with the exception of the Peronosporomycetes, which was represented by two Orders- Saprolegniales and Pythiales. In the filamentous fungal group, family Trichocomaceae was dominant followed by Saccharomycetaceae, Mucoraceae, Nectriaceae, Tremellaceae and Hypocreaceae. However, in the group of zoosporic & fungal like eukaryotes, family Saprolegniaceae was most dominant followed by Blastocladiaceae and Pythiaceae. A dramatic decrease in fungal load was observed in different seasons with highest colonial load in the summer season and lowest in the winter season. The observed distribution was statistically significant for both the filamentous fungal species (p < 0.01) as well as zoosporic fungi & fungal like eukaryotes (p < 0.05). In order to assess biodiversity patterns of fungi more accurately, it is necessary to repeat such investigations in other areas and to improve the tools for taxonomic identification of these highly diverse but mostly microscopic organisms.

A field survey on the status of gastrointestinal helminth parasites in hangul (Cervus elaphus hanglu) in Dachigam National Park of Kashmir.

One year crossectional survey was carried out to determine and describe the prevalence and intensity of gastrointestinal parasite infections in hangul (Cervus elaphus hanglu) in Dachigam National Park of Kashmir through faecal examinations. Out of 153 faecal samples examined, 82 (53.59 %) were found infected with GIT helminthes. In present study seven helminth species were found, including five nematode [Haemonchus contortus (55.39 %), Trichuris ovis (39.75 %), Dictyocaulus viviparus (28.4.00 %), Oesophogostomum circumcincta (13.7 %) and Chabertia ovina (4.02 %)] one trematode [Fasciola hepatica (17.3 %)] and one cestode species [Moneizia expansa (6.05 %)]. Based on the severity of infection 81.7 % of hangul positive samples were severely infected (epg > 1,500), 8.3 % heavily infected (epg = 1,100-1,500), 3.8 % moderately infected (epg = 800-1,000) and 7.2 % mildly infected (epg = 500). Season, sex and age were the factors that influenced the epidemiological prevalence of GIT helminths in hangul in the present study. The maximum helminth infection was observed in summer season and lowest in winter (P = 0.003). Lower age groups were more infected than adult animals (P > 0.05). Prevalence was higher in males than females (P > 0.05). The present study will initially be of great significance to add to existing knowledge of the epidemiology of GIT helminth of hangul which is the pioneering study on this animal in the valley and the findings will be quite helpful to devise the appropriate control and prophylactic strategies for GIT helminthiasis of hangul in the Dachigam national park.

Opportunistic fungi in lake water and fungal infections in associated human population in Dal Lake, Kashmir.

Natural habitats of opportunistic fungal pathogens are outside of the host; therefore, it is critically important to understand their ecology and routes of transmission. In this study, we investigated the presence of human pathogenic opportunistic fungi in lake water and incidence of fungal infections in associated population in Kashmir, India. Six hundred forty water samples were taken on seasonal basis from a wide network of sampling stations of the lake for an extended period of two years for screening their occurrence. The samples were inoculated onto rose bengal agar, malt extract agar, potato dextrose agar and other specified culture media supplemented with Chloramphenicol and Streptomycin followed by incubation at 37 °C. All the samples were positive for fungi, which were later identified by sequencing the rDNA internal transcribed spacer region aided by classical morphological culture techniques and physiological profiling. The whole process led to the isolation of sixteen species of opportunistic fungal pathogens belonging to genus Aspergillus, Candida, Penicillium, Cryptococcus, Fusarium, Rhizopus and Mucor in decreasing order of prevalence. Furthermore, 20% population (n = 384) of Dal inhabitants was examined for possible fungal infections and it was observed that only 8.07% individuals were positive for fungal infections with 4.68% skin infection cases, 2.34% onychomycosis cases and 1.04% candidiasis cases. Scrapings from onychomycosis and candidiasis patients showed the presence of Aversicolor and Calbicans respectively, resembling exactly the strains isolated from the lake water. However, the skin infection was because of a dermatophyte not isolated for the lake water. Higher prevalence of infection (6.77%) was seen in people using lake water followed by a positive prevalence of 1.30% using tap water. The results of present study suggest that the lake inhabitants are at a greater risk of getting life threatening fungal diseases which may lead to various morbidities.

Anthelmintic, antimicrobial, antioxidant and cytotoxic activity of Caltha palustris var. alba Kashmir, India.

The methanolic extract obtained from the root portion of Caltha palustris var. alba was evaluated for its anthelmintic efficacy against gastrointestinal nematodes of sheep under both in vitro and in vivo conditions using worm motility inhibition (WMI) assay and fecal egg count reduction (FECR) assay, respectively. The extract was subjected to antimicrobial activity using agar-well diffusion method against different bacterial strains. In addition the extract was evaluated for cytotoxic and antioxidant activity against cultured THP-1(Leukemia), A-549 (Lung), HCT-15 (Colon), Cervix (HeLa) and PC-3(Prostrate) cell lines by SRB and DPPH radical scavenging assays. The extract used resulted in mean %WMI of 94.44%, as observed when the worms were put in lukewarm buffer for 30 min after exposure to different treatments. The mean mortality index of the sample was 0.95. The lethal concentration (LC50) was 0.11 mg·mL(-1). Cell lines were exposed to concentration of 100 µg·mL(-1) of extract for 48 h, which reduced the viability of these cell lines. The same plant extract also showed 55.58% DPPH radical scavenging activity.

Antimicrobial and antioxidant activities of alcoholic extracts of Rumex dentatus L.

In-vitro antimicrobial and antioxidant activities of various concentrations ranging from 150 to 500 µg/ml of alcoholic (methanol and ethanol) extracts of Rumex dentatus were analyzed on different clinical bacterial strains (Shigella flexneri, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus) and fungal strains (Aspergillus versicolor, Aspergillus flavus, Acremonium spp., Penicillium dimorphosporum, Candida albicans, Candida kruesie, Candida parapsilosis) using agar disk diffusion method and broth dilution method (MIC and MBC determination) for antimicrobial activity and DPPH (1,1-diphenyl-2-picrylhydrazyl) assay, Riboflavin photo-oxidation assay, deoxyribose assay, lipid peroxidation assay for antioxidant activity. The extracts showed maximum inhibitory effect against K. pneumonia and P. aeruginosa with no activity against S. typhimurium from among the bacterial strains while as in case of the fungal strains the maximum effect was observed against C. albicans by both the extracts. MIC and MBC values determined for active fractions of the extracts against some bacterial strains (S. flexneri, K. pneumonia and E. coli) revealed that the test organisms were inhibited by all the extracts with methanol showing lower values of both MIC and MBC indicating it as a better antimicrobial agent. The antioxidant activity showed that the extracts exhibited scavenging effect in concentration-dependent manner on superoxide anion radicals and hydroxyl radicals leading to the conclusion that the plant has got a broad spectrum antimicrobial and antioxidant activity and could be a potential alternative for treating various diseases.

Anthelmintic and antimicrobial activity of methanolic and aqueous extracts of Euphorbia helioscopia L.

The aim of this study was to evaluate the anthelmintic and antimicrobial efficacy of Euphorbia helioscopia crude extracts. A worm motility inhibition assay and egg hatch assay were used for in vitro study, and a faecal egg count reduction assay was used for in vivo study. The in vitro study revealed anthelmintic effects of crude methanolic extracts of E. helioscopia on live Haemonchus contortus worms as evident from their paralysis and/or death at 8 h after exposure. Different concentrations (12.5 mg ml(-1), 25 mg ml(-1) and 50 mg ml(-1)) of aqueous and methanolic extracts were used against H. contortus which exhibited dose-dependent anthelmintic effects on H. contortus. Different extracts of E. helioscopia on percent inhibitory egg hatching was very low as compared to levamisole. The antimicrobial activity of extracts ranging from 100 to 500 mg ml(-1) screened by disc diffusion method against four selected bacterial (Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas multocida and Escherichia coli) and two fungal strains (Aspergillus flavus and Candida albicans) was also dose dependent with the extract showing more inhibitory effects against S. aureus and E. coli and a minimum inhibitory effect against A. flavus. It is concluded that the entire plant of E. helioscopia possesses significant anthelmintic and antimicrobial activity and could be a potential alternative for treating cases of helminth infections in ruminants.

In vitro and in vivo anthelmintic activity of Euphorbia helioscopia L.

The anthelmintic potentials of the aqueous and methanol extracts of Euphorbia helioscopia were investigated. In folklore medicine, E. helioscopia (Euphorbiaceae) is used in the treatment of various gastrointestinal ailments and diseases. A worm motility inhibition (WMI) assay and egg hatch assay (EHA) were used for in vitro assessment, and a faecal egg count reduction (FECR) assay was used for an in vivo study. The in vitro study revealed anthelmintic effects of crude methanolic extracts of E. helioscopia on live Haemonchus contortus worms as evident from their paralysis and/or death at 8h after exposure, different concentrations (12.5 mg ml(-1), 25 mg ml(-1) and 50 mg ml(-1)) of aqueous and methanolic extracts were used against H. contortus which exhibited dose-dependent anthelmintic effects on H. contortus. Aqueous and methanolic extracts of aerial parts (stem, leaves and flowers) of E. helioscopia were found to have very low percent inhibitory egg hatching as compared to levamisole. It is concluded that the entire plant of E. helioscopia possesses significant anthelmintic activity and could be a potential alternative for treating cases of helminth infections in ruminants.

Simple techniques to reduce bowel activity in cardiac SPECT imaging.

BACKGROUND AND AIM: Scatter from the bowel degrades image quality in 99mTc sestamibi myocardial perfusion imaging (MPI). Iodinated oral contrast, which has been used to outline bowel in medical imaging, absorbs X-rays as well as gamma rays. The purpose of this study was to test our hypothesis that iodinated oral contrast during MPI would absorb gamma rays emitted from 99mTc sestamibi in the bowel, thereby reducing scatter and improving cardiac SPECT images. METHODS AND RESULTS: Thirty subjects undergoing adenosine stress 99mTc sestamibi cardiac SPECT were randomized to receive either iodinated oral contrast (IOC), water or no intervention (controls). Subjects had 1 day rest-stress MPI using the adenosine stress protocol. Images were analysed using infra-cardiac counts, image variability, image contrast and the ratios of anterior to inferior and septal to lateral walls. The improvement in image contrast and variability between first and second images were significant in both the IOC and water groups. The IOC group had a more significant improvement in variability than did the water group. The reduction in infra-cardiac counts was also more significant in the IOC group. CONCLUSION: The use of oral contrast and water improved the image variability and contrast by decreasing the infra-cardiac scatter. The improvement was even more significant in the oral contrast group.