In vivo evaluation of new adjuvant systems based on combination of Salmonella Typhi porins with particulate systems: Liposomes versus polymeric particles.

Subunit vaccines that have weak immunogenic activity require adjuvant systems for enhancedcellular and long-acting humoral immune responses. Both lipid-based and polymeric-based particulate adjuvants have been widely investigated to induce the desired immune responses against the subunit vaccines. The adjuvant efficacy of these particulate adjuvants depends upon their physicochemical properties such as particle size, surface charge, shape and their composition. Previously, we showed in vitro effect of adjuvant systems based on combination of chitosan and Salmonella Typhi porins in microparticle or nanoparticle form, which were spherical with positive surface charge. In the present study, we have further developed an adjuvant system based on combination of porins with liposomes (cationic and neutral) and investigated the adjuvant effect of both the liposomal and polymeric systems in BALB/c mice using a model antigen, ovalbumin. Humoral immune responses were determined following priming and booster dose at 15-day intervals. In overall, IgM and IgG levels were induced in the presence of both the liposomal and polymeric adjuvant systems indicating the positive impact of combination with porins. The highest IgM levels were obtained on Day 8, and liposomal adjuvant systems were found to elicit significantly higher IgM levels compared to polymeric systems. IgG levels were increased significantly after booster, particularly more profound with the micro-sized polymeric system when compared to cationic liposomal system with nano-size. Our results demonstrated that the developed particulate systems are promising both as an adjuvant and delivery system, providing enhanced immune responses against subunit antigens, and have the potential for long-term protection.

Effects of injectable trace element and vitamin supplementation during the gestational, peri-parturient, or early lactational periods on neutrophil functions and pregnancy rate in dairy cows.

The aim of this study was to evaluate effects of injectable trace element and vitamin combination on phagocytic, oxidative burst activity of neutrophils and reproductive outcomes in dairy cows. Cows were to assigned to the following groups: (1) injectable trace element supplementation (ITES, n = 44, containing zinc, manganese, copper, selenium); (2) injectable vitamin supplementation (VIT, n = 48, containing vitamins A, D3, E); (3) ITES + VIT (n = 46); and (4) control (CON, n = 44). Cows were administered four injections between 230 and 260 days of the gestational period, on day of parturition, and 30 days postpartum. Neutrophil function was assessed at 10 days before and after calving. Phagocytosis was greater in cows of the ITES + VIT group at 10 days prepartum (P < 0.05) while oxidative burst was similar among groups. There were greater non-esterified fatty acid (NEFA) concentrations in cows of the ITES+VIT group at 10 days prepartum (P < 0.05). Cows supplemented with ITES+ VIT had less SOD activity than those supplemented with ITES or vitamin during the pre- to post-partum transition period (P < 0.05). The total odds of pregnancy were greater in cows supplemented with trace element and/or vitamin (P < 0.05). In conclusion, supplementation of ITES and/or VIT resulted in an increased total pregnancy rate. Vitamin or trace element supplementation did not differ with the control group in both the prepartum and postpartum period for immune variables. There, however, was greater phagocytosis in cows supplemented with vitamin and trace elements during the prepartum period that might be related to metabolic-induced inflammation.

Development and in vitro evaluation of a new adjuvant system containing Salmonella Typhi porins and chitosan.

In order to improve the immunogenicity of the highly purified vaccine antigens, addition of an adjuvant to formulation, without affecting the safety of the vaccine, has been the key aim of the vaccine formulators. In recent years, adjuvants which are composed of a delivery system and immunopotentiators have been preferred to induce potent immune responses. In this study, we have combined Salmonella Typhi porins and chitosan to develop a new adjuvant system to enhance the immunogenicity of the highly purified antigens. Cationic gels, microparticle (1.69 ± 0.01 µm) and nanoparticles (337.7 ± 1.7 nm) based on chitosan were prepared with high loading efficiency of porins. Cellular uptake was examined by confocal laser scanning microscopy, and the macrophage activation was investigated by measuring the surface marker as well as the cytokine release in vitro in J774A.1 macrophage murine cells. Porins alone were not taken up by the macrophage cells whereas in combination with chitosan a significant uptake was obtained. Porins-chitosan combination systems were found to induce CD80, CD86 and MHC-II expressions at different levels by different formulations depending on the particle size. Similarly, TNF-α and IL-6 levels were found to increase with porins-chitosan combination. Our results demonstrated that combination of porins with chitosan as a particulate system exerts enhanced adjuvant effect, suggesting a promising adjuvant system for subunit vaccines with combined immunostimulating activity.

Impact of Prepartum Body Condition Score Loss on Metabolic Status During the Transition Period and Subsequent Fertility in Brown Swiss Dairy Cows.

INTRODUCTION: The objectives of this study were to determine the role of a fall in pre-calving body condition score (BCS) in postpartum metabolic status and reproductive outcomes, and gauge the indicativeness of blood metabolites during the transition period. MATERIAL AND METHODS: Cows were grouped based on BCS loss between days -14 ±3 and 0 relative to calving. Cows that lost no BCS were the BCS control group (BCS-C), cows that lost 0.25 BCS points the low BCS loss group (BCS-L), and those that lost 0.5 points or more the high BCS loss (BCS-H) group. Blood was taken on days -14 ±3, 3, 14, and 30 ±4 for determination of comprehensive metabolic panel biomarker levels. Beta-hydroxybutyric acid (BHBA) levels were quantified on postpartum examination days. Vaginal discharge scores, ovarian activity on day 30 ±4, and subsequent fertility parameters were evaluated. RESULTS: The BCS-H cows had lower mean Ca concentrations before calving and on day 3, when the group's BHBA and CK were higher (P < 0.05); on day 14 they had higher AST concentrations (P < 0.05). The BCS-L cows had greater bilirubin levels (P < 0.05). The BCS-H cows had lower cyclicity and higher endometritis rates. First service pregnancy rates were 50%, 50%, and 61.9%, open days 96.8, 95.75, and 89.2, and overall pregnancy rates 56.25%, 65%, and 80.95 % in the BCS-H, BCS-L, and BCS-C groups, respectively. CONCLUSION: Prepartum BCS loss of ≥ 0.5 points could be associated with Brown Swiss cow low Ca and BHBA concentrations early postpartum, and with subsequent uterine health and overall pregnancy rate. Prepartum Ca concentration might be a prognostic biomarker for postpartum metabolic status and reproductive outcomes.

The evaluation of superoxide dismutase activity, neutrophil function, and metabolic profile in cows with retained placenta.

The present study aimed to determine the alterations in pre- and postpartum immune function, superoxide dismutase (SOD) activity, and serum metabolite changes in Brown Swiss cows with or without retained placenta (RP). Peripheral blood was sampled at -10 ±â€¯4 days antepartum and 10 ±â€¯4 days postpartum for the assessment of neutrophil phagocytic and oxidative burst activities by using flow cytometry. SOD activity, beta hydroxybutiric acid (BHB), nonesterified fatty acids (NEFA), calcium (Ca), phosphorus, aspartate aminotransferase (AST), alkaline phosphatase (ALP), alanine aminotransferase (ALT), creatinine kinase (CK), gamma-glutamyl transferase (GGT), lactate dehydrogenase, bilirubin, cholesterol, glucose, urea, albumin, total protein, and triglyceride concentrations were also measured from serum samples collected at -10 ±â€¯4, 3 ±â€¯1, 10 ±â€¯4, and 30 ±â€¯4 days relative to parturition. All data were analyzed using the MIXED procedure of SPSS. Neutrophil phagocytic and oxidative burst activities, phagocytic and oxidative fluorescence intensity, and phagocytic and oxidative indexes were similar among the groups (P > 0.05). However, serum SOD activity was 30 ±â€¯4 IU/ml and 17 ±â€¯2 IU/mL for cows that subsequently developed RP and healthy cows at -10 ±â€¯4 days antepartum, respectively (P < 0.05). Body condition scores were similar in both groups (P > 0.05). All investigated serum metabolic parameters varied significantly over time (P < 0.001). Cows affected with RP had subclinical hypocalcemia (8.06 ±â€¯0.3 mg/dl) and had significantly higher BHB levels (1027 ±â€¯137.9 mmoL/l) at 3 ±â€¯1 days postpartum. Furthermore, these cows had greater AST and NEFA concentrations around calving (P < 0.05). In conclusion, retained placenta had no prolonged effect on the measures of neutrophil function during the postpartum period. On the other hand, increased antepartum SOD activity might be a predisposing factor and can be used as a predictive biomarker for retained placenta.

Capsaicin inhibits cell proliferation by cytochrome c release in gastric cancer cells.

Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide) is the principal pungent component in hot peppers. The role of capsaicin in carcinogenesis is quite controversial. Although some investigators suspect that capsaicin is a carcinogen, co-carcinogen, or tumor promoter, others have reported that it has chemopreventive and chemotherapeutic effects. The present study aimed to evaluate the cytotoxicity and chemosensitizing activities of capsaicin alone and on 5-flourouracil (5-FU)-treated gastric cancer cells. In this study, the gastric cancer cell line HGC-27 was used and capsaicin used as a chemosensitizer and 5-flourouracil (5-FU) was used as chemotherapeutic. Cytotoxicity and chemosensitizing activities were analyzed with MTT assay; supernatant levels of LDH and glucose were detected as biochemical markers of cell viability; cytochrome c and AIF were evaluated with western blot; and additionally, wound-healing assays were employed. Results suggested that capsaicin had significant anticancer abilities; such capsaicin were capable of causing multifold decreases in the half maximal inhibitory concentration IC50 value of 5-FU. The continuing controversy surrounding consumption or topical application of capsaicin clearly suggests that more well-controlled epidemiologic studies are needed to evaluate the safety and efficacy of capsaicin use. In summary, the present study demonstrated that capsaicin has the potential to be used for treating gastric carcinoma with 5-FU in vitro.

Identification of hypoglycaemic compounds from berries of Juniperus oxycedrus subsp. oxycedrus through bioactivity guided isolation technique.

ETHNOPHARMACOLOGICAL RELEVANCE: Decoction of Juniperus oxycedrus subsp. oxycedrus L. (Cupressaceae) berries is used internally as tea and pounded fruits are consumed to lower blood glucose levels in Turkey. AIM OF THE STUDY: To evaluate hypoglycaemic and antidiabetic activity of J. oxycedrus subsp. oxycedrus berries and to identify active compounds through bioactivity guided isolation technique. MATERIAL AND METHODS: Hypoglycaemic effect of J. oxycedrus subsp. oxycedrus (Joso) berry extracts on oral administration was studied using in vivo models in normal, glucose-hyperglycaemic rats. Streptozotocin induced diabetic rats were used to examine antidiabetic activity of Joso extracts, subextracts, fractions, subfractions and shikimic acid (SA). RESULTS: Through in vivo bioactivity-guided fractionation processes, shikimic acid, 4-O-ß-d-glucopyranosyl ferulic acid and oleuropeic acid-8-O-ß-d-glucopyranoside were isolated from the n-butanol subextract by silica gel and reverse phase column chromatography as the main active ingredient of the active subfraction. After 8 days administration of the major compound shikimic acid, blood glucose levels (24%), malondialdehyde levels in kidney tissues (63-64%) and liver enzymes (AST, ALT, ALP) of diabetic rats were decreased. CONCLUSION: Results indicated that Joso berry extract and its active constituents might be beneficial for diabetes and its complications.