Combination of ascorbate and alpha-tocopherol as a preventive therapy against structural and functional defects of erythrocytes in visceral leishmaniasis

The redox unbalance in erythrocytes has been found to contribute significantly in the development of anemia in visceral leishmaniasis (VL). The present study revealed enhanced production of reactive oxygen species (ROS) and gradual depletion of alpha-tocopherol and ascorbate in the erythrocytes of infected animals. The response of erythrocytes to chronic treatment with antioxidants was studied in hamsters during leishmanial infection. Treatment with a combination of alpha-tocopherol and ascorbate proved to be the most effective preventive for the proteolytic degradation of erythrocyte membrane. Erythrocytes from infected animals were thermally more sensitive compared to the control ones. Combination of both antioxidants was most successful in resisting heat induced structural defects in the cells. Cross-linking of membrane proteins subsequent to oxidative damage in the red cells was accompanied by the formation of high molecular weight protein band at the top of the resolving gel in the presence of the cross-linking agent dimethyladepimidate (DMA). Marked inhibition of cross-linking was observed with combination of both antioxidants. Treatment with alpha-tocopherol and ascorbate together could withstand osmotic lysis of erythrocytes in the infected animals very efficiently. Decreased hemoglobin (Hb) level was successfully replenished and was coupled with significant increase in the life span of red cells after treating the animals with both antioxidants. Results indicate better efficacy of the combination therapy with alpha-tocopherol and ascorbate in protecting the erythrocytes from structural and functional damages during leishmanial infection.     

Immunogenicity and protective efficacy of three DNA vaccines encoding pre-erythrocytic- and erythrocytic-stage antigens of Plasmodium cynomolgi in rhesus monkeys

Although several malaria vaccine candidate antigens have been identified, the most suitable methods for their delivery are still being investigated. In this regard, direct immunization with DNA encoding these vaccine target antigens is an attractive alternative. Here, we have investigated the immune responses to DNA immunization with three major vaccine target antigens: the apical membrane antigen-1 and the 19-kDa C-terminal fragment of merozoite surface protein-1 from the erythrocytic stage, and the thrombospondin-related adhesive protein from the pre-erythrocytic stage of Plasmodium cynomolgi in rhesus monkeys. Antigen-specific antibodies were developed in all the immunized monkeys and peripheral blood mononuclear cells from all immunized monkeys proliferated to different extents upon in vitro stimulation with the corresponding recombinant proteins. The immunized monkeys were challenged with P. cynomolgi sporozoites. All of the immunized animals developed infection but although there was no significant difference between the control and vaccinated animals in terms of pre-patent period, total duration of patency and primary peak parasitemia, the vaccinated animals had significantly lower secondary peak parasitemia than the control animals.     

Structural and immunological characterization of recombinant ovomucoid expressed in Escherichia coli

The expression of recombinant allergens is becoming new insights of an important diagnosis and the therapy of allergies as well as molecular approaches to immunological and structural studies of allergens. Ovomucoid is a major food allergens in the hen's egg white which causes immediate food-hypersensitivity reactions mainly in children. A gene coding for the cDNA representing an entire ovomucoid molecule has been cloned in Escherichia coli under the control of T5 promoter fused with six-Histidine tag at the amino terminal end. Upon induction, the E. coli cells, harbouring this construct, expressed the recombinant protein as a soluble fraction and the recombinant ovomucoid protein was purified to electrophoeretic homogeneity using Ni²⁺ nitrilotriacetic acid agarose affinity chromatography. Immunoblot analysis showed that human IgE and IgG binding activities of the recombinant ovomucoid was identical to that of native analogue. The antigenicity and allergenicity of recombinant ovomucoid were almost same as that of native form when tested with an ELISA using six individual patient's serum. CD spectra indicated that that the recombinant ovomucoid has more -helix and less -structure than native form. These results show that the recombinant ovomucoid constructed in this study could be used for further studies on the immunological and structural studies of ovomucoid.     

Effects of farmyard manure and phosphorus on zinc transformations and phyto-availability in two alfisols of India

Laboratory pot experiments were run to study the effects of added zinc (Zn) with and without farmyard manure (FYM) and phosphorus (P) on Zn transformations in two Alfisols, together with Zn uptake by wheat plants grown up to 60 days. In the first experiment the treatments included four levels of Zn (0, 3.75, 7.5 and 15 mgkg(-1) soil) and two levels of FYM (0 and 10 tha(-1)), and in the second experiment five levels of P (0, 20, 40, 80 and 160 mgkg(-1) soil) and one level of Zn (7.5 mgkg(-1) soil). The soils were sequentially fractionated into water-soluble plus exchangeable (CA-Zn), inorganically bound (AAC-Zn), organically bound (PYR-Zn), oxide bound (OX-Zn) and residual (RES-Zn) forms. The effect of added FYM was more evident on the OX-Zn fraction and the percentage utilization of Zn by wheat was the greatest with the addition of FYM alone at the rate of 10 tha(-1) (1.95-2.38%) in comparison to other treatment combinations. Among the levels, application of 7.5 mg Zn kg(-1) soil showed the maximum increase in different fractions of soil Zn and significantly increased the Zn utilization by wheat (0.87-1.17%) as compared to other Zn levels (0.58-0.88%). On an average, about 85% of the added Zn was recovered in different fractions in Zn treated pots. However, the recovery per cent of the added Zn was significantly higher at Zn level 7.5 (95%) mgkg(-1) soil than at 3.75 (87%) and 15 (73%) mg Zn kg(-1) soil levels. Phosphorus additions up to 40 mgkg(-1) soil increased the plant-available Zn in soils whereas at higher P levels plant-available forms decreased with a concominant increase in the inert forms. At 160 mg P kg(-1) soil, the P effect was more pronounced in the shoot than in the root, suggesting that a higher P level inhibits Zn translocation from root to upper plant parts. Path analysis showed that the organically (PYR-Zn) and inorganically bound (AAC-Zn) Zn fractions were the predominant fractions that influenced the Zn availability to plants.     

Nuclear gene trees and the phylogenetic relationships of the mangabeys (Primates: Papionini)

Phylogenetic relationships of mangabeys within the Old World monkey tribe Papionini are inferred from analyses of nuclear DNA sequences from five unlinked loci. The following conclusions are strongly supported, based on congruence among trees derived for the five separate gene regions: (1) mangabeys are polyphyletic within the Papionini; (2) Cercocebus is the sister taxon to the genus Mandrillus; and (3) Lophocebus belongs to a clade with Papio and Theropithecus, with Papio as its most likely sister taxon. Morphologically based phylogenies positing mangabey monophyly were evaluated by mapping the sequences for each locus on these trees. The data seem to fit these trees poorly in both maximum-parsimony and likelihood analyses. Incongruence among nuclear gene trees occurred in the interrelationships among Lophocebus, Papio, and Theropithecus. Several factors that may account for this incongruence are discussed, including sampling error, random lineage sorting, and introgression.      

PPCMatrix: a PowerPC dotmatrix program to compare large genomic sequences against protein sequences

Summary : An interactive dotmatrix program for the MacOS was designed that allows comparison of DNA to protein sequences using nested 3-frame translations. Availability : Shareware, available at http://copan.bioz.unibas.ch/software/ Contact : burglin@ubaclu. unibas.ch      

Alcohol Use among Adolescent Youth: The Role of Friendship Networks and Family Factors in Multiple School Studies

To explore the co-evolution of friendship tie choice and alcohol use behavior among 1,284 adolescents from 12 small schools and 976 adolescents from one big school sampled in the National Longitudinal Study of Adolescent to Adult Health (AddHealth), we apply a Stochastic Actor-Based (SAB) approach implemented in the R-based Simulation Investigation for Empirical Network Analysis (RSiena) package. Our results indicate the salience of both peer selection and peer influence effects for friendship tie choice and adolescent drinking behavior. Concurrently, the main effect models indicate that parental monitoring and the parental home drinking environment affected adolescent alcohol use in the small school sample, and that parental home drinking environment affected adolescent drinking in the large school sample. In the small school sample, we detect an interaction between the parental home drinking environment and choosing friends that drink as they multiplicatively affect friendship tie choice. Our findings suggest that future research should investigate the synergistic effects of both peer and parental influences for adolescent friendship tie choices and drinking behavior. And given the tendency of adolescents to form ties with their friends'' friends, and the evidence of local hierarchy in these networks, popular youth who do not drink may be uniquely positioned and uniquely salient as the highest rank of the hierarchy to cause anti-drinking peer influences to diffuse down the social hierarchy to less popular youth. As such, future interventions should harness prosocial peer influences simultaneously with strategies to increase parental support and monitoring among parents to promote affiliation with prosocial peers.     

Evaluation of the impact of shielding materials in radiation protection in transgenic animals

We are using a plasmid-based transgenic mouse mutation model system to evaluate the effectiveness of aluminum or low-density polyethylene (LDPE) shielding after 250 MeV/u protons or 1 GeV/u iron ion irradiation. Transgenic mice, with multiple copies of the plasmid pUR288 lacZ transgene integrated into the genome of every cell of the animal, were either irradiated or sham-treated. Multiple endpoints, including early cytogenetic damage in erythrocytes at 48 h after exposure, chromosome aberrations in bone marrow lymphocytes, and lacZ mutant frequencies (MF) in brain and spleen tissues were measured in the same animals. The frequency of total circulating reticulocytes (fRET) dropped precipitously at 48 h after 2 Gy of proton irradiation. The average level of micronucleated reticulocytes (fMN–RET) was fivefold higher in the irradiated samples relative to the controls at the same time point. There was an increase in total chromosome aberrations in bone marrow lymphocytes at 8 weeks after proton irradiation but this increase was not statistically significant relative to the controls. Evaluation of the lacZ MF in the brain and spleen tissues showed that proton irradiation induced a twofold increase in MF in each tissue. Similar samples were collected from animals that were shielded from the proton beam by aluminum. Compared to the unshielded treatment group, we noted no difference in fRET, fMN–RET, chromosome aberrations in lymphocytes and lacZ MF in brain and spleen tissues obtained from these animals. In a separate study, animals were exposed to high-energy iron ions with or without 10 or 15 cm LDPE. Using the same approach, we noted a precipitous drop in fRET, and an elevation in fMN–RET within 48 h after 1 Gy of iron ions. Total chromosome aberrations in bone marrow lymphocytes were slightly elevated but not significant at 8 weeks after iron ion exposure. Shielding animals with 10 or 15 cm of polyethylene appeared to have no effect on the level of RET, MN–RET or chromosome aberrations in these animals. LacZ MF in brain and spleen tissues increased 1.5–2-fold above control levels after 1 Gy iron ions at 8 weeks after treatment. On the other hand, MF in tissues harvested from shielded animals appeared to be lower than their unshielded litermates, suggesting the polyethylene shielding was effective in reducing the iron-induced genomic damage in tissues. Although shielding may be effective, in some cases, in reducing the physical dose of particle radiation, our cytogenetic results showed that the biological impact of the particle beam remain unchanged. On the other hand, reduction in transgene MF in tissues from LDPE-shielded animals but not in the aluminum-shielded animals strongly suggests that careful consideration of the biological endpoints used is necessary in the evaluation of the efficacy of the selected shielding material.     

Phosphorus availability to rice (Oriza sativa L.)-wheat (Triticum estivum L.) in a Vertisol after eight years of inorganic and organic fertilizer additions

Integrated use of inorganic fertilizer N and well decomposed cattle manure (CM) or 30-35 days old Parthenium (Parthenium hysterophorus L.), a weed grown off site as green manure (GM) under repeated applications of fertilizer P and urea N for eight years in a rice (Oriza sativa L.)-wheat (Triticum estivum L.) sequence was studied on transformation of fertilizer P applied to soil at the National Research Center for Weed Science, Jabalpur, India. Based on the results, it appeared that, repeated applications of 52 kg super-phosphate P resulted in a marked increase in Olsen P linearly with time. Conjunctive use of urea fertilizer N with organic manure resulted in a larger increase in Olsen P in the Vertisol. Studies further revealed that the greater accumulation of fertilizer P applied in excess to crop removal occurred in inorganic P in the plots receiving only fertilizer N. However, plots receiving fertilizer N along with organic manures led to P accumulation predominantly in organic forms. The study suggests that these two pools of P acted as a sink when fertilizer P was applied in excess to crop removal and are bio-chemically active. The Olsen P status after 8 cycles of rice-wheat crops revealed that the average amount of fertilizer P required after adjusting for crop uptake to increase Olsen P by 1 mg kg(-1) soil was 7.2 kg Pha(-1) in the plots receiving only fertilizer N. Whereas, application of 5t FYM or 6t GM reduced it to 4.6 kg Pha(-1). The plots receiving manure always maintained a greater concentration of Olsen P. The application of CM or GM with fertilizer N enriched short-term inorganic P as well as long-term organic P fertility. After eight years, larger concentrations of organic P in the subsurface layer (16-30 cm), compared to initial values, indicates downward movement of P in organic forms.