Evaluation of therapeutic potential of recombinant buffalo lactoferrin N-lobe expressed in E. coli.

Lactoferrin (Lf) is a multifunctional bi-lobate iron-binding glycoprotein belonging to transferrin family with a mass of approximately 80 kD. Being ubiquitously present in almost all biological secretions, it performs important biological functions. One of the earliest and very well-documented functions of Lf is the antibacterial effect against broad spectrum Gram-negative and Gram-positive bacteria. In this study, buffalo Lf N-lobe cDNA was amplified, cloned and expressed as a fusion protein in Escherichia coli cells using pQE30 expression vector. After post-induction confirmation of expressed protein by SDS-PAGE, purification of recombinant protein using Ni-NTA was attempted and the yield of recombinant buffalo N-lobe Lf was estimated to be 1 mg/ml. Antibacterial activity of recombinant buffalo Lf N-lobe was assessed on pathogenic E. coli and Staphylococcus aureus strains. Peptic digest of recombinant N-lobe buffalo Lf showed antibacterial activity comparable to commercially available bovine Lf. The successful expression and characterization of functional recombinant N-lobe of buffalo Lf expressed in E. coli opens new vistas for developing alternate therapeutics, particularly against the diseases caused by Gram-negative microbes such as septicemia and diarrhea in newborn calves and mastitis in dairy animals.

Differential expression of cytokines in PBMC of Bos indicus and Bos taurus × Bos indicus cattle due to Brucella abortus S19 antigen.

Brucellosis is the most dreadful disease among bovines, although breed differences have been observed in prevalence of disease, worldwide. In present study, antibody response and relative expression of proinflammatory cytokines was compared in Bos indicus (zebu) and Bos taurus × Bos indicus (crossbred) cattle vaccinated by live attenuated Brucella abortus S19 antigen. Six female calves (4-6 months age) of both groups were vaccinated with B.abortus S19 strain. Blood samples were collected before vaccination (0d) and 7th (7d), 14th (14d) and 28th (28d) days after vaccination. Indirect ELISA showed high (p < .05) anti-Brucella antibody level after vaccination; with no significant difference between the groups. During Real-time expression, IFNγ, TNFα, IL6 and IL10 genes initially showed down regulation followed by upregulation in both the groups; however, the trend was much prominent in crossbreds. The expressions of IFNγ, TNFα and IL6, proinflammatory molecules important for initial containment of the Brucella were significantly (p < .01) higher in crossbred. The study showed that the Sahiwal cattle were less responsive to B.abortus S19 antigen than crossbreds, indicating its lower sensitivity to the Brucella, comparatively. In contrary, higher expression of the proinflammatory molecules in crossbreds could be important for containment of the organism during initial stage of infection.

Eating and weight concerns among Sikkimese adolescent girls and their biocultural correlates: an exploratory study.

OBJECTIVES: Growing concern about ideal body image among adolescent girls in developing countries has led them to follow dietary modifications. In general, the ideal body image is perceived as a 'thin body' and now tagged with the concept of being physically fit. The present study evaluates the prevalence of weight concerns, and subsequent eating behaviour modifications among a group of adolescent girls in Sikkim. DESIGN: Pre-tested FFQ and dietary behaviour questionnaire were canvassed through direct interviews. Anthropometric measurements were taken following standard techniques. In the present study, dieting stands for skipping meals or avoiding certain food items. SETTING: Sikkim, India. SUBJECTS: A total of 577 girls were selected from several blocks of all four districts of Sikkim. RESULTS: The results of the study show that concern with weight reduction is growing among adolescent girls, particularly among urban girls of affluent families. Girls from families with a higher economic status are about two times more likely to report dissatisfaction with their body weight (OR = 1·96; P ≤ 0·05) and these girls are five times more likely to report the need for dieting. CONCLUSIONS: It is evident that weight concern and dissatisfaction over body weight are growing among adolescents. It can be safely argued that eating disorder, once a problem of the Western world, is now slowly creeping in among adolescents of the developing world as a consequence of rapid lifestyle changes over the past few decades.