The discovery and biochemical characterization of the secretory pathway Ca 2؉ -ATPase, PMR1, in Saccharomyces cerevisiae, has paved the way for identification of PMR1 homologues in many species including rat, Caenorhabditis elegans, and Homo sapiens.
We have solubilized and purified the histidine-tagged yeast secretory pathway/Golgi ion pump Pmr1 to near homogeneity in one step, using nickel affinity chromatography.
Leishmaniasis remains a major health problem of the tropical and subtropical world. The visceral form causes the most fatalities if left untreated. Dramatic increases in the rates of infection and drug resistance and the non-availability of safe vaccines have highlighted the need for identification of novel and inexpensive anti-leishmanial agents. This study reports that racemoside A, a water-soluble steroidal saponin purified from the fruits of Asparagus racemosus, is a potent anti-leishmanial molecule effective against antimonial-sensitive (strain AG83) and -unresponsive (strain GE1F8R) Leishmania donovani promastigotes, with IC 50 values of 1.15 and 1.31 mg ml "1 , respectively. Incubation of promastigotes with racemoside A caused morphological alterations including cell shrinkage, an aflagellated ovoid shape and chromatin condensation. This compound exerts its leishmanicidal effect through the induction of programmed cell death mediated by the loss of plasma membrane integrity as detected by binding of annexin V and propidium iodide, loss of mitochondrial membrane potential culminating in cell-cycle arrest at the sub-G 0 /G 1 phase, and DNA nicking shown by deoxynucleotidyltransferase-mediated dUTP end labelling (TUNEL). Racemoside A also showed significant activity against intracellular amastigotes of AG83 and GE1F8R at a 7-8-fold lower dose, with IC 50 values of 0.17 and 0.16 mg ml "1 , respectively, and was non-toxic to murine peritoneal macrophages up to a concentration of 10 mg ml "1 . Hence, racemoside A is a potent anti-leishmanial agent that merits further pharmacological investigation.
Extracts were prepared from leaf, bark, and seed of Azadirachta indica, leaf and seed of Prunus persica, bark of Mangifera indica, and leaf of Psidium guajava and were evaluated against Boophilus microplus. Of the eight extracts screened, the extracts prepared from the A. indica seed showed very high level of efficacy (80%) after 5 h of treatment. Besides the immediate effect on adult ticks, the egg-laying properties of the survived ticks was also assessed, and a significant reduction (P < 0.01) in the reproductive index of ticks fed on animals treated with A. indica seed extracts was noted in comparison to control. The efficacy of the neem seed extracts was compared with the commonly used synthetic pyrethroids, and comparable efficacy against B. microplus fed on animals treated with neem seed extracts and acaricide treated was noted. The possibility of using the extracts in IPM format for the management of ticks is discussed.
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