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Objectives: It is essential to obtain homogeneous drug mixtures, especially when only a fraction of the prepared dose is to be administered. This study aimed to validate a manual mixing method for guaranteeing homogeneity., Methods: One operator tested six standardised manual mixing techniques (one, five and 10 inversions, and one, five and 10 bottoms-up agitations) six times each for preparations in bags and syringes. The mixing step was reproduced experimentally by adding a small volume of analyte (0.8 mL for syringes and 6 mL for bags) to a large volume of matrix (50 mL for syringes and 300 mL for bags). Three analyte/matrix pairings were tested: water/water, water/glucose 20% and glucose 20%/water. The tracer (sodium chloride) was assayed using capillary electrophoresis. Volume measurement errors were corrected by weighing bags and syringes. In order to evaluate inter-individual variability, the 10 inversions technique was tested by 10 drug preparation technicians. Mixtures were considered acceptable if they were between 95% and 105% accurate and if the coefficient of variation was ≤5% of the average of the six repetitions., Results: Both the 10 inversions and 10 bottoms-up agitations mixing techniques ensured acceptable mixtures by the principal technician in all tested conditions. When mixing using the 10 inversions method was tested by the 10 technicians, the mixture's mean acceptability could no longer be ensured., Conclusion: Use of a standardised mixing technique did not appear to be sufficient to obtain a homogeneous mixture across technicians. Standardised guidelines for needle position, needle rinsing and speed of addition should be implemented., Competing Interests: Competing interests: None declared.

Objective: To evaluate the effect of oral administration of Kombucha tea (K-tea) on lead induced oxidative stress., Methods: Sprague Dawley rats were administered 1 mL of 3.8% lead acetate solution daily alone or in combination with K-tea orally for 45 d, and the antioxidant status and lipid peroxidation were evaluated., Results: Oral administration of lead acetate to rats enhanced lipid peroxidation and release of creatine phosphokinase and decreased levels of reduced glutathione (GSH) and antioxidant enzymes (superoxide dismutase, SOD and glutathione peroxidase, GPx). Lead treatment did not alter humoral immunity, but inhibited DTH response when compared to the control. Lead administration also increased DNA fragmentation in liver. Oral administration of Kombucha tea to rats exposed to lead decreased lipid peroxidation and DNA damage with a concomitant increase in the reduced glutathione level and GPx activity. Kombucha tea supplementation relieved the lead induced immunosuppression to appreciable levels., Conclusion: The results suggest that K-tea has potent antioxidant and immunomodulating properties.

Objective: The objective of the study was to evaluate toxicity, anti-stress activity and hepato-protective properties of Kombucha tea., Method: Kombucha tea was fed orally for 15 days using three different doses i.e. normal dose, five and ten times the dose. Rats were then sacrificed and various biochemical, and histological parameters were estimated. Anti-stress activity was evaluated either by 1) by exposing animals to cold and hypoxia and estimating the levels of malondialdehyde and reduced glutathione in plasma/blood or 2) by subjecting the animals to restraint stress and recording faecal output. Hepato-toxicity was induced by challenging the animals to an acute dose of paracetamol (1 gm/kg) orally and determining the plasma levels of SGPT, SGOT and MDA., Results: The effect of oral administration of different doses of K-tea to albino rats was examined and the results indicate that K-tea has no significant toxicity as revealed by various biochemical and histopathological parameters. K-tea has been found to prevent lipid peroxidation and fall in reduced glutathione level when rats were exposed to cold and hypoxia in simulated chamber. Further, K-tea has also been found to decrease the Wrap-restraint faecal pellet output in rats. K-tea has also been found to decrease paracetamol induced hepatotoxicity significantly., Conclusion: The study shows that K-tea has anti-stress and hepato-protective activities.

Gamma interferon (IFN-gamma)-secreting CD4+ T cells have long been established as an essential component of the protective immune response against Mycobacterium tuberculosis. It is now becoming evident from studies with the murine model of tuberculosis that an important role also exists for major histocompatibility complex (MHC) class I-restricted CD8+ T cells. These cells are capable of acting as both IFN-gamma secretors and cytotoxic T lymphocyte (CTL) effectors; however, their exact role in immunity against tuberculosis remains unclear. This study demonstrates the presence of Mycobacterium bovis BCG-reactive CD8+ T cells in healthy BCG-vaccinated donors and that these CD8+ T cells are potent cytokine producers as well as cytotoxic effector cells. Using FACScan analysis, we have shown that restimulation with live M. bovis BCG induced more CD8+-T-cell activation than the soluble antigen purified protein derivative and that these cells are actively producing the type 1 cytokines IFN-gamma and tumor necrosis factor alpha (TNF-alpha). These CD8+ T cells also contain the cytolytic granule perforin and are capable of acting as potent CTLs against M. bovis BCG-infected macrophages. The mycobacterial antigens 85A and B (Ag85A and Ag85B, respectively), and to a lesser extent the 19- and 38-kDa proteins, are major antigenic targets for these mycobacterium-specific CD8+ T cells, while whole-M. bovis BCG activated effector cells from these BCG-vaccinated donors, as expected, failed to recognize the 6-kDa ESAT-6 protein. The use of metabolic inhibitors and blocking antibodies revealed that the CD8+ T cells recognize antigen processed and presented via the classical MHC class I pathway. These data suggest that CD8+ T cells may play a critical role in the human immune response to tuberculosis infection.

The effect of exposure to hypoxia on macrophage activity, lymphocyte function and oxidative stress was investigated. Hypoxia enhanced peritoneal macrophage activity as revealed by enhanced phagocytosis and free radical production. There was no significant change in antibody titres to sheep red blood cells in either serum or spleen during hypoxia. However, there was a considerable reduction in the delayed-type hypersensitivity response to sheep red blood cells, indicating the impairment of T-cell activity. Hypoxia decreased the blood glutathione (reduced) level and increased plasma malondialdehyde by a factor of about 2. It is therefore speculated that hypoxia imposes an oxidative stress leading to decreased T-cell acivity.