Anticancer and anti-inflammatory activities of some dietary cucurbits.

In this study, we investigated few dietary cucurbits for anticancer activity by monitoring cytotoxic (MTT and LDH assays), apoptotic (caspase-3 and annexin-V assays), and also their anti-inflammatory effects by IL-8 cytokine assay. Aqua-alcoholic (50:50) whole extracts of cucurbits [Lagenaria siceraria (Ls), Luffa cylindrica (Lc) and Cucurbita pepo (Cp)] were evaluated in colon cancer cells (HT-29 and HCT-15) and were compared with isolated biomolecule, cucurbitacin-B (Cbit-B). MTT and LDH assays revealed that the cucurbit extracts and Cbit-B, in a concentration dependent manner, decreased the viability of HT-29 and HCT-15 cells substantially. The viability of lymphocytes was, however, only marginally decreased, yielding a potential advantage over the tumor cells. Caspase-3 assay revealed maximum apoptosis with Ls while annexin V assay demonstrated maximum efficacy of Lc in this context. These cucurbits have also shown decreased secretion of IL-8, thereby revealing their anti-inflammatory capability. The results have demonstrated the therapeutic potential of dietary cucurbits in inhibiting cancer and inflammatory cytokine.

Free radical scavenging and metal chelation by Tinospora cordifolia, a possible role in radioprotection.

Aqueous extract of T. cordifolia inhibited Fenton (FeSO4) reaction and radiation mediated 2-deoxyribose degradation in a dose dependent fashion with an IC50 value of 700 microg/ml for both Fenton and radiation mediated 2-DR degradation. Similarly, it showed a moderate but dose dependent inhibition of chemically generated superoxide anion at 500 microg/ml concentration and above with an IC50 value of 2000 microg/ml. Aqueous extract inhibited the formation of Fe2+-bipiridyl complex and formation of comet tail by chelating Fe2+ ions in a dose dependent manner with an IC50 value of 150 microg/ml for Fe2+-bipirydyl formation and maximally 200 microg/ml for comet tail formation, respectively. The extract inhibited ferrous sulphate mediated lipid peroxidation in a dose-dependent manner with an IC50 value of 1300 microg/ml and maximally (70%) at 2000 microg/ml. The results reveal that the direct and indirect antioxidant actions of T. cordifolia probably act in corroboration to manifest the overall radioprotective effects.

Protective effect of RH-3 with special reference to radiation induced micronuclei in mouse bone marrow.

Effect of pre-irradiation administration of different doses of RH-3, the herbal preparation of an Indian medicinal plant Hippophae rhamnoides, 30 min before 10 Gy whole body gamma irradiation was studied. Doses between 25 to 35 mg/kg body wt. were found to render > 80 % survival in mice. In order to investigate whether RH-3 protected against radiation induced genotoxicity, mice were administered different doses of RH-3, 30 min before 2 Gy dose and compared with untreated, RH-3 treated and irradiated controls. The bone marrow cells were collected at different time intervals following various treatments and processed for scoring micronuclei (MN). Administration of RH-3 alone did not enhance the MN frequency as compared to the control, and radiation dose of 2 Gy significantly enhanced the MN frequency (3.1 %, P < 0.01). Pre-irradiation treatment with RH-3, however, reduced the radiation induced MN frequency in a drug dose dependent manner suggesting its radioprotective efficacy. The protective effect of RH-3 on radiation induced perturbations in cell cycle progression was studied flowcytometrically in mouse bone marrow cells. RH-3 treatment (30 mg/kg body wt.) enhanced DNA synthesis (S-phase) in unirradiated controls and also countered radiation induced depression of S-phase to facilitate replenishment of cells lost due to radiation injury.

Iron chelation and related properties of Podophyllum hexandrum, a possible role in radioprotection.

Aqueous extract of Podophyllum species has been reported to render significant protection against radiation induced mortality, cytogenetic damage and cell death. In view of this, present study was undertaken to investigate its antioxidant properties. Chelation, oxidation and reduction of Fe2+ and Fe3+ were measured using chelating agents 2-2' bipiridyl and potassium thiocyanate respectively. Podophyllum extract, in a dose dependent manner, chelated Fe2+ more efficiently than Fe3+ and also modulated Fe2+/Fe3+ ratio. Homogenate of mouse liver was used to measure TBARS for estimating lipid peroxidation. Podophyllum extract also inhibited lipid peroxidation in a dose dependent manner and maximum inhibition (92%) was achieved at 1000 micrograms/ml concentration. These results demonstrates that Podophyllum exhibits antioxidant properties as seen through chelation and modulation of redox state of iron ions and these may primarily contribute towards its radioprotective manifestation.

Antitumour and radioprotective action of Podophyllum hexandrum.

A significant antitumour effect of P. hexandrum, a herb thriving at Himalayas (2500-4000 m), was observed in strain 'A' mice carrying solid tumours developed by transplanting Ehrlich ascites tumour (EAT). Subtoxic well tolerated sequential doses of aqueous extract of P. hexandrum (a daily dose of 34.5 mg/kg b.w. for 15 days) enhanced tumour doubling time (TDT) from 1.94 +/- 0.26 days to 19.1 +/- 2.5 days. However, no synergism was revealed between radiation and P. hexandrum, though both independently manifested antitumour effects. In normal mice, pre-irradiation administration of extract of P. hexandrum protected mice in a dose dependent manner (optimal dose being 34.5 mg/kg body.wt. rendering 72% survival for 30 days) against whole body lethal irradiation of 10 Gy. Radioprotective properties of P. hexandrum were found to be comparable to synthetic radioprotectors like diltiazem etc.

Prevention of radiation induced taste aversion in rats.

Diltiazem, a calcium channel blocker, and a cardiovascular therapeutic agent offers significant protection to mice against lethal dose of ionizing radiation. Considering the potential efficacy of diltiazem as a radioprotector for human use, it was deemed necessary to investigate its influence on radiation-induced behavioural changes like nausea, vomiting, learning, memory and performance. In the present studies, conditioned taste aversion (CTA) test based on consumption of saccharin solution, was used as a marker of behavioural changes. Significant CTA (97 +/- 2%) was observed in rats irradiated with Co-60 gamma rays (absorbed dose 1 Gy). Administration of diltiazem at doses greater than 10 mg/kg, body wt, evoked CTA in a dose-dependent manner and that was found to be further aggravated on irradiation. At a lower dose of 5 mg/kg, body wt, diltiazem did not evoke CTA and protected against radiation induced aversion significantly (62 +/- 3%). The results suggest that diltiazem at concentrations lower than 10 mg/kg, body wt, in rats may be useful in preventing radiation induced behavioural changes. This observation could be of particular significance in clinical radiotherapy where radiation induced nausea and vomiting are of great concern.

Radioprotective effects of diltiazem on cytogenetic damage and survival in gamma ray exposed mice.

Diltiazem, a calcium ion channel blocker, already in use in cardiovascular therapeutics, has been observed to protect against bone marrow damage (cytogenetic damage, cell death) and mortality in whole body irradiated mice. The micronuclei fraction in bone marrow cells of whole body irradiated (60Co gamma rays, 2.0 Gy) mice was reduced from 2.24 +/- 0.23% to about 0.74 +/- 0.33% by preirradiation administration (-20 min) of 110 mg/kg body wt. diltiazem (ip). Endogenous colony forming unit counts in spleen of mice administered 110 mg/kg body wt. (-20 min) of diltiazem before 10 Gy whole body irradiation were 6 times more than untreated irradiated controls. Pretreatment with diltiazem accelerated the recovery of radiation induced weight loss also. Diltiazem (110 mg/kg body wt, -20 min) enhanced 30 day survival to about 95% and 85% after lethal whole body absorbed dose of 9 and 10 Gy respectively and also mitigated radiation induced life- span shortening. Post-irradiation (10 Gy) administration of diltiazem (+20 to 30 min) enhanced survival from about 2 to 15% only but was highly significant (P < 0.001). Possible modes of radioprotective action of diltiazem have been discussed.

Some naturally occurring phytophototoxins for mosquito control.

Alpha-terthiophene (alpha-T) and erythrosin-B, the naturally occurring plant secondary metabolites, were tried for their phototoxic properties against Anopheles and Culex larvae under dark, ordinary tube light (1.9-2.4 w/m2) and sun light (680-840 w/m2). LC50 values of alpha-T for Anopheles larvae (4th instar) were found to be 154, 92 and 11 ppb under dark, tube light and sunlight, respectively. For Culex larvae corresponding LC50 values under different light conditions were 129, 97 and 22 ppb. Erythrosin-B under all photoregimens was found to be less toxic to larvae of both Anopheles and Culex sps. Also, the susceptibility of the mosquito species decreased with age, towards alpha-T and erythrosin-B. Cumulative effects in terms of delay in metamorphosis were also observed among survivors of such exposures. The effects of these compounds were also seen on the adults and developing unhatched embryos of a common aquatic snail (Lymnaea sps). The LC50 values of alpha-T for adults were found to be 39, 23 ppm and 77 ppb under dark, tube light and sunlight and for developing unhatched embryos the corresponding values were 620, 41 and 13 ppb. Erythrosin-B was found to be much less toxic under sunlight and dark, to both adults and embryos as compared to the toxicity of alpha-T. Potential use of such biodegradable and eco-friendly compounds of natural origin in mosquito control is discussed.

A rapid method for flow cytometric cell counting and cell cycle analysis from monolayer cultures of tumor cells.

A rapid, simple, and reliable flow cytometric method using the histochemical fluorescent stain Hoechst 33342 in presence of the non-ionic detergent Triton X-100 has been reported. The processing of melanoma cell cultures to get nuclei stained with the fluorescent dye was accomplished in one step and within an hour permitted concurrent flow cytometric measurement of cell density and cell cycle analysis. The preparation is stable for more than three weeks at room temperature for flow cytometry. The histograms are reproducible and exhibit a coefficient of variation of less than 2.5% (G1 peak). The cell density measurements varied within +/- 5% limits.

Inhibition of skin tumors in DMBA-induced complete carcinogenesis system in mice by garlic (Allium sativum).

The chemopreventive action of garlic extract on DMBA-induced complete skin carcinogenesis system was studied in random bred, 6-7 weeks old, male Swiss albino mice. Topical weekly application of DMBA for 25 weeks at two dose levels, i.e. 200 nmol during the first week followed by 100 nmol during subsequent weeks, or 400 nmol during the first week followed by 200 nmol during subsequent weeks, resulted in 73.9% and 100% tumor incidences respectively. When garlic extract was topically applied twice daily for 3 days every week prior to above-stated dose schedules of DMBA, the incidences of tumors were reduced to 31.8% (P less than 0.01) and 43.4% (P less than 0.01) respectively.