Study of glucose uptake activity of Helicteres isora Linn. fruits in L-6 cell lines

The effect of hot water extract of fruits of Helicteres isora on glucose uptake was studied in rodent skeletal muscle cells (L-6 cells) involved in glucose utilization. H. isora is an antidiabetic medicinal plant being used in Indian traditional medicine. Hot water extracts were analysed for glucose uptake activity and found to be significantly active at 200 μg/ml dose comparable with insulin and metformin. Elevation of glucose uptake by H. isora in association with glucose transport supported the upregulation of glucose uptake. It was concluded that hot water extract of H. isora activate glucose uptake in L-6 cell line of mouse skeletal muscles

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Not AvailableVelvet bean is an important medicinal legume, its seeds are prominent source of L-Dopa. The present investigation on genetic diversity assessment of 58 germplasm of velvet bean by using 11 ISSR markers. Out of 63 amplified products 59 were showed polymorphism and 4 were monomorphic with an average of 5.7 bands amplified per primer. According to band statistics and efficiency parameters showed the primers UBC 827, UBC 834 and UBC 836 were more efficient. The highest genetic similarity values (0.90) were observed between IIHR MP 102 and IIHR MP 74-3. In dendrogram germplasm grouped into two major clusters at 63 per cent similarity. Among the germplasm, IIHR Selection 4, IIHR Selection 10, IIHR MP 9, IC 33243 and IIHR MP 7 were found to be distinctly divergent, can be used in the further breeding programme.Not Availabl

Th2 cytokine mRNA expression.

<p>Th2 cytokines mRNA expression profile analysis of normal and immunized hamsters on days 45 p.c.and 90 p.c were checked by quantitative real-time RT-PCR. The expression of Th2 cytokines viz. IL4 (a,b), IL10 (c,d), TGFß (e,f) was also observed to be ~3.00 folds higher (p<0.001) in the Infected hamsters as compare to vaccinated group as well as control groups. Significance values indicate the difference between the vaccinated group and infected group (***, p<0.001), vaccinated group and BCG group (***, p<0.001), vaccinated group and normal group (*, p<0.05).</p

Parasite burden.

<p>Clinical outcomes following <i>L</i>. <i>donovani</i> challenge in hamsters immunized with rLdSir2RP+BCG. On day 21 after the booster, the hamsters of infected, BCG alone and vaccinated groups were challenged intracardially with 10^<b>7</b> metacyclic promastigotes of <i>L</i>. <i>donovani</i>. Following parameters observed Body weight (a), spleen weight (b), liver weight (c), Parasite burden (no. of amastigotes per 100 cell nuclei) in the spleen(d), liver(e), and bone marrow (f) on days 45 and 60 p.c. Significance values indicate the difference between the vaccinated groups and infected group (***, p<0.001). Data represent mean values standard errors (SE) at the designated time points.</p

Leishmania-specific IgG and its isotypes IgG1 and IgG2 responses.

<p>Serum samples were collected from different groups of hamsters at designated time points and assayed for rLdSir2RP—specific IgG, IgG1, and IgG2 levels by ELISA. Antibody responses in rLdSir2RP vaccinated hamsters in comparison to the unimmunized infected hamsters on days 45, 60 and 90 p.c. The IgG1 were observed to be elevated by 1 to 2 fold in infected control group in comparison to the group of immunized hamsters with rLdSir2RP at the time interval of Days 45(a), d60(b) and d90(c). There was significant elevation of IgG2 level (by 3.5 to 4 folds) in group of hamsters immunized with rLdSir2RP at the time interval of Days 45(a), d60(b)and d90(c). Data are presented as the absorbance at 492 nm and are means ± SE for 3–4 hamsters per group in triplicate wells. Significance values indicate the difference between the vaccinated group and infected group (***, p<0.001).</p

Expression of recombinant LdSir2RP in E. coli Rosetta strain and its purification.

<p>Expression and purification of rLdSir2RP (1A, IB) in prokaryotic cells, Whole Cell Lysate (WCL) of transformed <i>E</i>. <i>coli</i> separated on 12% acrylamide gel and stained with Coomassie blue, M: Molecular wt. markers; Lane 1: WCL before IPTG induction; lane 2: WCL after IPTG (1.0 mM) induction at 37°C. Lane 5: eluted protein (1B). Western blot analysis using anti-rLdSir2RPAb in uninduced WCL, induced WCL and SLD—M: Mol wt marker, Lane 1: uninduced WCL, Lane 2: induced WCL (1C).</p

Cellular responses of rLdSir2RP of L.donovani in vaccinated hamsters.

<p>Nitric oxide production (μM) (a, b, c) by J774A.1 cell line. The J774 A.1 cells were primed with the supernatants of stimulated lymphocytes (3 days with mitogen and 5 days with Ags) of normal, infected and vaccinated hamsters in response to rLdSir2RP, SLD and LPS respectively at 10 μg/ml each. The estimation of NO production was done using Greiss reagent in supernatants collected from macrophage cultures 24 h after incubation and the absorbance of the reaction product was measured at 540 nm. XTT response (d, e, f) of mononuclear cells (lymph nodes) from normal, <i>L</i>. <i>donovani</i> infected and vaccinated hamsters in response to Con A, SLD and rLdSir2RP at 10 μg/ml each Proliferation was represented as mean OD. The data represent the means of triplicate wells ± S.D. Significance values indicate the difference between the SLD and rLdSir2RP stimulation (**, p < 0.01; ***, p<0.001).</p

Discovery of a New Class of Natural Product-Inspired Quinazolinone Hybrid as Potent Antileishmanial agents

The high potential of quinazolinone containing natural products and their derivatives in medicinal chemistry led us to discover four novel series of 53 compounds of quinazolinone based on the concept of molecular hybridization. Most of the synthesized analogues exhibited potent leishmanicidal activity against intracellular amastigotes (IC₅₀ from 0.65 ± 0.2 to 7.76 ± 2.1 μM) as compared to miltefosine (IC₅₀ = 8.4 ± 2.1 μM) and nontoxic toward the J-774A.1 cell line and Vero cells. Moreover, activation of Th1 type and suppression of Th2 type immune responses and induction in nitric oxide generation proved that <b>8a</b> and <b>8g</b> induce murine macrophages to prevent survival of parasites. Compounds <b>8a</b> and <b>8g</b> exhibited significant in vivo inhibition of parasite 73.15 ± 12.69% and 80.93 ± 10.50% against Leishmania donovani/hamster model. Our results indicate that compounds <b>8a</b>, <b>8g</b>, and <b>9f</b> represent a new structural lead for this serious and neglected disease

Synthesis, Structure–Activity Relationships, and Biological Studies of Chromenochalcones as Potential Antileishmanial Agents

Antileishmanial activities of a library of synthetic chalcone analogues have been examined. Among them, five compounds (<b>11</b>, <b>14</b>, <b>16</b>, <b>17</b>, <b>22</b>, and <b>24</b>) exhibited better activity than the marketed drug miltefosine in in vitro studies against the intracellular amastigotes form of Leishmania donovani. Three promising compounds, <b>16</b>, <b>17</b>, and <b>22</b>, were tested in a L. donovani/hamster model. Oral administration of chalcone <b>16</b>, at a concentration of 100 mg/kg of body weight per day for 5 consecutive days, resulted in >84% parasite inhibition at day 7 post-treatment and it retained the activity until day 28. The molecular and immunological studies revealed that compound <b>16</b> has a dual nature to act as a direct parasite killing agent and as a host immunostimulant. Pharmacokinetics and serum albumin binding studies also suggest that compound <b>16</b> has the potential to be a candidate for the treatment of the nonhealing form of leishmaniasis