Traditional uses of medicinal plants for dermatological healthcare management practices by the Tharu tribal community of Uttar Pradesh, India

The aim of present study was to explore and document medicinal plants used for the traditional dermatological healthcare management practices by the the Tharu tribal community of Uttar Pradesh. The study was conducted during 2000–2004. Information was gathered from 230 informants residing in 46 villages in Terai region of Indo-Nepal boarder using questionnaires; oral interviews and group discussions. Total 92 medicinal plant species were cited for the preparation of 113 crude drug formulations. Voucher specimens of cited plant species were collected and identified as belonging to 82 genera and 49 families. Thirty-nine medicinal plant species were reported for the first time for dermatological healthcare problems from India. The dermatological healthcare problems managed were cut and wounds, ringworm, leprosy, eczema, scabies, leucoderma, boils, carbuncles, pimples, skin blemishes, spots, eruption, and burns etc. The most commonly and popularly used medicinal plant species for management of dermatological healthcare problems in the study area were Curcuma longa L., Azadirachta indica A. Juss and Melia azedarach L. It is concluded that dermatological healthcare management practice in the study area depends largely on wildly growing medicinal plant species. There is an urgent need to properly conserve the medicinal plant species growing in this area for human welfare. There is also need for further phytopharmacological studies to provide scientific explanation for the usages of 57 medicinal plant species for which to the best of our knowledge phytopharmacological literatures are not available.     

Cadmium Enhances Generation of Hydrogen Peroxide and Amplifies Activities of Catalase, Peroxidases and Superoxide Dismutase in Maize

Maize (Zea mays L. cv. 777) plants grown in hydroponic culture were treated with 50 μm CdSO₄. Growth and metabolic parameters indicative of oxidative stress and antioxidant responses were studied in leaves of plants treated with Cd. Apart from increasing lipid peroxidation and H₂O₂ accumulation, supply of Cd suppressed growth, fresh and dry mass of plants and decreased the concentrations of chloroplastic pigments. The activities of catalase (CAT; EC 1.11.1.6), peroxidase (POD; EC 1.11.1.7), ascorbate peroxidase (APX; EC 1.11.1.11) and superoxide dismutase (SOD; EC 1.15.1.1) were increased in plants supplied 50 μm Cd. Localization of activities of isoforms of these enzymes (POD, APX and SOD) on native gels also revealed increase in the intensities of pre‐existing bands. Stimulated activities of CAT, POD, APX and SOD in maize plants supplied excess Cd do not appear to have relieved plants from excessive generation of reactive oxygen species (ROS). It is, therefore, concluded that supply of 50 μm Cd induces oxidative stress by increasing production of ROS despite increased antioxidant protection in maize plants.     

Amelioration of Sodic Soil for Wheat Cultivation Using Bioaugmented Organic Soil Amendment

A novel bioaugmented organic amendment (SF_OA) [consisted of vermicompost (pre‐enriched with plant growth promoting fungi) mixed with pressmud and Azadirachta indica (A. Juss.) seed cake] was developed to reclaim sodic soil and support wheat production. A field trial of the SF_OA application with/without chemical fertilizers conducted in completely randomized design with four replications to compare growth, yield and seed protein contents of wheat (Triticum aestivum L.) on sodic soil. The favourable changes occurred in different properties of amended soils were studied. A combined application of chemical fertilizer and SF_OA significantly (p < 0·05) increased the number of spikelet per plant (63%) and weight of grains per ear (65%) in the amended soil compared with the control. Likewise, the grain yield, weight of 1000 grains and seed protein contents of wheat were significantly (p < 0·05) increased in the combined application compared with other treatments. The expression of protein bands with molecular weights of 36, 52 and 66 kDa were higher in seeds of wheat under the combined treatment. The improvement in wheat production was attributed to significant favourable changes in different soil characteristics such as bulk density, total organic C, alkaline phosphatase, β‐glucosidase, dehydrogenase and cellulase activities that were increased by 234%, 181%, 234%, 176%, 189% and 150%, respectively, in case of amended soil under the combined treatment compared with the control. The tested SF_OA may be recommended as soil amendment for reclaiming sodic soil and supporting wheat cultivation with better crop growth and yield in combination with chemical fertilizers on sodic soil. Copyright © 2014 John Wiley & Sons, Ltd.     

Induction of protective immune response in mice by a DNA vaccine encoding Trypanosoma evansi beta tubulin gene

Surra, caused by Trypanosoma evansi, is an economically important veterinary disease of the tropics. Lack of effective drugs or vaccines have made surra a severe economic burden particularly in Asia and sub-Saharan Africa. In this study, a naked DNA construct encoding full length T. evansi beta (β) tubulin gene was used to immunize mice, to elicit a T. evansi β tubulin protein specific humoral immune response, delineated by ELISA. The serum cytokine profile post immunization, as determined by flow cytometry bead based assay, showed a predominant T helper cell Type 1 (Th1) response with significant increase in levels of IFNγ and TNFα. Lethal challenge with T. evansi blood-form trypomastigotes post immunization generated a β tubulin specific recall response and a stronger Th1 type serum cytokine profile which correlated with an extended survival and better control of parasitemia in the immunized mice.     

A multilaboratory evaluation of a commercial enzyme-linked immunosorbent assay test for the detection of antibodies against Mycobacterium avium subsp. paratuberculosis in cattle.

Five laboratories participated in a study to evaluate sources of variation in results from an enzyme-linked immunosorbent assay (ELISA) for antibodies against Mycobacterium avium subsp. paratuberculosis. Each laboratory repeatedly tested duplicates of a negative, positive (P), and high-positive (HP) serum sample, which were supplied by the United States Department of Agriculture: Animal and Plant Health Inspection Service: Veterinary Services, National Veterinary Services Laboratories, Ames, IA, on all 96-well microtiter plates when routinely testing other samples for M. avium subsp. paratuberculosis antibodies. These 3 sera were aliquoted and sent to the 5 participating laboratories. This study focused on variation in test results because of assay reagents and laboratory techniques and did not account for biologic variability associated with the time course of infection in cattle. Overall, results from 868 microtiter plates were used in the study. For each sample a sample-to-positive (S/P) ratio was calculated according to the manufacturer's directions. The S/ P ratio for the P sample ranged from 0.06 to 1.039 (mean = 0.466 and 0.484 for wells 1 and 2, respectively) and those for the HP sample ranged from 2.446 to 8.727 (mean = 4.027 and 3.980 for wells 1 and 2, respectively). The majority of the variation in S/P ratio for the P sample was attributed to kit lot (37.5%), followed by random (unexplained) error (27.0%), laboratory (18.3%), and kit lot by laboratory (11.9%). By eliminating plates in which the separation between negative and positive control ODs was less than 0.4, the proportion of variation attributed to laboratory was reduced markedly. These results confirm that there is variability in M. avium subsp. paratuberculosis ELISA results and that several sources contribute to the observed variability. The study gives a relative estimate of the contribution of various sources to the overall variability observed in the M. avium subsp. paratuberculosis ELISA results with kit lot being a primary contributor. Similar data for other ELISA tests for antibodies to M. avium subsp. paratuberculosis or other antigens also should be developed.     

Quantification of seedborne infection by Rhynchosporium secalis in barley using competitive PCR

The potential of the competitive polymerase chain reaction (PCR) assay for quantification of seedborne infection by Rhynchosporium secalis in barley was examined using a primer set (RS1 and RS3) derived from the internal transcribed spacer (ITS) regions of ribosomal RNA genes of this pathogen. Introduction of a heterologous internal control, which competes for the same primer set in the conventional PCR assay, allowed for detection and quantification of R. secalis fungal biomass. In order to generate a standard calibration curve, DNA prepared from infected seeds with different levels of R. secalis infection was subjected to competitive PCR assay. The resulting PCR product ratio for each PCR reaction ( R. secalis -amplified DNA/internal control template-amplified DNA) increased proportionally with increasing levels of infected seed DNA in the reaction mixture. Naturally infected seed lots collected from 1995 to 1999 were used to demonstrate the potential of the competitive PCR assay as an alternative seed health testing method. The results from this competitive PCR assay were compared with those from conventional visual disease assessment and an agar plate assay. Although relatively good correlation between visual disease assessment and the competitive PCR was found in the case of artificially mixed seed samples, there was poor correlation in the experiments using naturally infected seed samples.     

High-level expression of SAG1 and GRA7 gene of Toxoplasma gondii (Izatnagar isolate) and their application in serodiagnosis of goat toxoplasmosis

Toxoplasmosis, caused by Toxoplasma gondii, is a disease of economic importance in livestock, especially in sheep and goats, where it causes abortion. Although several serological tests are in use for diagnosis of infection, production of reliable reagents is a constraint. An 814 bp sequence coding for a truncated surface antigen surface antigen 1 (SAG1), a tachyzoite stage-specific protein, as well as a 657 bp sequence coding for granule protein 7 (GRA7), a dense granule protein were PCR amplified from the genomic DNA of T. gondii. The amplified products were ligated in pET-32b(+) and pET-32c(+) expression vectors, respectively and subsequently transformed into BL21(DE3)pLysS cells. A high-level expression of the histidine-tagged SAG1 and GRA7 fusion proteins were obtained after 7h of incubation. The recombinant proteins were purified using Ni-NTA column and were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis using reference positive sera from goat, rabbit and humans at 1:100 dilution. Subsequently, the diagnostic efficiency of the recombinant proteins, either individually or as a cocktail of the recombinant proteins, was assessed with 56 reference goat sera by enzyme-linked immunosorbent assay (ELISA). The immunoreactivity of the refolded SAG1 and GRA7 was evidenced by high OD values. The reactivity of the recombinant proteins as a cocktail preparation was more than that of individual proteins in ELISA and could detect accurately the infection in goats. This is the first report of serological detection of caprine toxoplasmosis by ELISA using a cocktail of recombinant Toxoplasma proteins.     

Functional genomic analysis of RNA interference in C. elegans

RNA interference (RNAi) of target genes is triggered by double-stranded RNAs (dsRNAs) processed by conserved nucleases and accessory factors. To identify the genetic components required for RNAi, we performed a genome-wide screen using an engineered RNAi sensor strain of Caenorhabditis elegans. The RNAi screen identified 90 genes. These included Piwi/PAZ proteins, DEAH helicases, RNA binding/processing factors, chromatin-associated factors, DNA recombination proteins, nuclear import/export factors, and 11 known components of the RNAi machinery. We demonstrate that some of these genes are also required for germline and somatic transgene silencing. Moreover, the physical interactions among these potential RNAi factors suggest links to other RNA-dependent gene regulatory pathways.     

Live attenuated malaria vaccine designed to protect through hepatic CD8⁺ T cell immunity

Our goal is to develop a vaccine that sustainably prevents Plasmodium falciparum (Pf) malaria in ≥80% of recipients. Pf sporozoites (PfSPZ) administered by mosquito bites are the only immunogens shown to induce such protection in humans. Such protection is thought to be mediated by CD8(+) T cells in the liver that secrete interferon-γ (IFN-γ). We report that purified irradiated PfSPZ administered to 80 volunteers by needle inoculation in the skin was safe, but suboptimally immunogenic and protective. Animal studies demonstrated that intravenous immunization was critical for inducing a high frequency of PfSPZ-specific CD8(+), IFN-γ-producing T cells in the liver (nonhuman primates, mice) and conferring protection (mice). Our results suggest that intravenous administration of this vaccine will lead to the prevention of infection with Pf malaria.