Objective: This study aimed to examine the effect of sodium butyrate (SB) on growth performance, immune status, organs weights, and microarchitecture of lymphoid organs and small intestine. Methods: A total of 120, 1-d-old broiler chicks were distributed into the following four treatment groups: corn-soy based basal diet (BD) without supplement (control), or the same BD supplemented with 0.1 g/kg zinc bacitracin (ZnB), 0.5 g/kg SB (SB-0.5), or 1.0 g/kg SB (SB-1), respectively. Six birds/group were killed on d-21 and d-35, and samples were collected. Results: Cell-mediated immune response at 48 h post-Phytohemagglutinin-P injection, and antibody titer against Newcastle disease vaccine and sheep red blood cells on d-35 was noted higher (p<0.05) in SB-1 compared to ZnB and control. Lower (p<0.05) feed conversion ratio (FCR) was attained by the supplemented groups. Thymus and spleen weighed more (p<0.05) in SB-1, and bursa registered more (p<0.05) weight in both SB groups compared to control. On d-21, areas of thymus medulla and spleen germinal centers were noted higher (p<0.05) in SB-1 group. The villus height and villus surface area increased (p<0.05) in duodenum and jejunum in both SB groups on d-21, and in SB-1 on d-35, respectively compared to ZnB and control. On d-21, number of goblet cells containing mucins of acidic nature increased (p<0.05) in all the segments of small intestines in SB-1 group compared to control, and on d-35 in ileum compared to other groups. Conclusion: In conclusion, SB improved growth performance and immunity as well as modulated morphology of lymphoid organs and gut mucosa in broiler chickens.
This study was conducted to investigate the effects of brown seaweed (Undaria pinnatifida) by-product and seaweed fusiforme (Hizikia fusiformis) by-product supplementation on growth performance and blood profiles including serum immunoglobulin (Ig) in broilers. Fermentation of seaweeds was conducted by Bacillus subtilis and Aspergillus oryzae. In a 5-wk feeding trial, 750 one-d-old broiler chicks were divided into 5 groups, and were assigned to the control diet or experimental diets including control+0.5% brown seaweed (BS) by-product, control+0.5% seaweed fusiforme (SF) by-product, control+0.5% fermented brown seaweed (FBS) by-product, and control+0.5% fermented seaweed fusiforme (FSF) by-product. As a consequence, body weight gain (BWG) and gain:feed of seaweed by-product groups were clearly higher, when compared to those of control diet group from d 18 to 35 and the entire experimental period (p<0.05). In mortality rate, seaweed by-product groups were significantly lower when compared to control diet group during entire experimental period (p<0.05). However, Feed Intake of experimental diets group was not different from that of the control group during the entire experimental period. Whereas, Feed Intake of fermented seaweed by-product groups was lower than that of non-fermented seaweed groups (p<0.05). Total organ weights, lipids, and glutamic oxalacetic transaminase (GOT) of all treatment groups were not different from those of control group. However, glutamic pyruvate transaminase (GPT) of all treatment groups was higher than that of control group at d 17 (p<0.05). In case of serum Igs concentration, the concentration of IgA antibody in BS, SF, FSF treatment groups was significantly higher than in control group at d 35 (p<0.01). IgA concentration in FBS supplementation groups was negligibly decreased when compared to the control group. IgM concentration in the serums of all treatment groups was significantly higher than in control group (p<0.05) and in fermented seaweed by-product groups were much higher than in non-fermented seaweed groups (p<0.05). On the other hand, IgG concentrations in all treatment groups were lower than in control group (p<0.05). Taken together, our results suggest that by-product dietary supplementation of BS, SF, FBS, and FSF in poultry may provide positive effects of growth performance and immune response.
Heat shock protein (HSP) expression is unregulated in tumor cells and, HSP expression is likely marker of the malignant potential of oral epithelial lesion. Furthermore, the 70kDa HSP is implicated in the degree of tumor differentiation, the rate of tumor proliferation and the magnitude of the anti-tumor immune response. Accordingly, the distribution and intensity of HSP 70 and HSP 47 expression was assessed in the DMBA induced oral carcinogenesis in hamster. Golden Syrian hamsters which were 3 months-age and 90-120g were collected. 9,10-dimethyl-1,2-benzanthracene (DMBA) in a 0.5% solution in mineral oil was painted on the buccal pouch mucosa 3 times per week in the study group. In each control and experimental groups of 6, 8, 10, 12, 14, 16, 18, 20 weeks, specimen were sectioned for immunohistochemical study with anti-HSP47 and anti-HSP70 antibody. The following results were obtained. 1. HSP47 positive cells were rare or negative of normal oral mucosa, increased mildly in basal and suprabasal basal layer, and spinous cell layer after experimental 6 weeks (dysplastic or CIS stage). In CIS stage, HSP47 expression is prominent in dysplastic free or normal adjacent epithelium. 2. HSP 47 positive cells in connective tissue were mainly inflammatory cells, which is gradually increased from control to precancerous and cancer stage. But HSP47 positive cells after 14 weeks were decreased, especially normal and cancer adjacent epithelium. 3. The positive staining cells of HSP70 in control, dysplastic, and CIS stage were not seen. But they were mild findings in basal layer and moderate findings in spinous layer after experimental 14 weeks (cancer stage). 4. HSP70 positive cells were increased in precancerous and cancer stage than control group in connective tissue. After experimental 16 weeks, we could not find the HSP expression in cancer cells according to cancer differentiation or cancer stage. It is concluded that HSP70 or HSP47 expression is not a definitive marker of oral malignancy or malignant potential. However, with further development, HSP immunoreactivity may be valuable as an adjunct to conventional histology for assessing the malignant potential of oral mucosal lesions.
Lee, Hye-Jeong;Kim, Ji Young;Park, Ji Eun;Yoon, Yong-Dal;Tsang, Benjamin K.;Kim, Jong-Min
Development and Reproduction
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v.20
no.4
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pp.315-329
/
2016
Fas ligand (FasL) and its receptor Fas have been implicated in granulosa cell apoptosis during follicular atresia. Although interferon-gamma (IFN-${\gamma}$) is believed to be involved in the regulation Fas expression in differentiated granulosa or granulosa-luteal cells, the expression of this cytokine and its role in the regulation of the granulosa cell Fas/FasL system and apoptosis during follicular maturation have not been thoroughly investigated. In the present study, we have examined the presence of IFN-${\gamma}$ in ovarian follicles at different stage of development by immunohistochemistry and related their relative intensities with follicular expression of Fas and FasL, and with differences in granulosa cell sensitivity to Fas activation by exogenous agonistic Anti-Fas monoclonal antibody (Fas mAb). Although IFN-${\gamma}$ immunostaining was detectable in oocyte and granulosa cells in antral follicles, most intense immunoreactivity for the cytokine was observed in these cells of preantral follicles. Intense immunoreactivity for IFN-${\gamma}$ was most evident in granulosa cells of atretic early antral follicles where increased Fas and FasL expression and apoptosis were also observed. Whereas low concentrations of IFN-${\gamma}$ (10-100 U/mL) significantly increased Fas expression in undifferentiated granulosa cells (from preantral or very early antral follicles) in vitro, very higher concentrations (${\geq}1,000U/mL$) were required to up-regulate of Fas in differentiated cells isolated from eCG-primed (antral) follicles. Addition of agonistic Fas mAb to cultures of granulosa cells at the two stages of differentiation and pretreated with IFN-${\gamma}$ (100 U/mL) elicited morphological and biochemical apoptotic features which were more prominent in cells not previously exposed to the gonadotropin in vivo. These findings suggested that IFN-${\gamma}$ is an important physiologic intra-ovarian regulator of follicular atresia and plays a pivotal role in regulation of expression of Fas receptor and subsequent apoptotic response in undifferentiated (or poorly differentiated) granulosa cells at an early (penultimate) stage of follicular development.
The role of passive cell-mediated transfer of immunity against primary amoebic meningoen- cephalitis(PAME) in mice was studied. Waegleria fowleri, ITMAP 359, were cultured in CGVS medium. The ICR mice used were six week-old males of average weight of 15 g. Immunization was done by three intraperitoneal injections of $1{\times}10^6$ N. fowleri trophozoites at the interval of one week. Splenocytes were obtained from normal and immune mice spleens, and Ix107 cells were administered intraperitoneally into mice 3 days before challenge infection. Mice were infected intranasally with $7{\times}10^4$ N. fowleri trophozoites in a $3{\;}{\mu}l$ suspension under secobarbiturate anesthesia. Transplants of normal or immune splenocytes seem to alter the pattern of the PAME level- opment. The splenocytcs transferred from immune mice reduced the mortality rate in the JV. fowleri infected mice, as compared with the mice transferred with the same number of normal splenocytes or without splenocyte, The blastogenic response of the splenocytes to both lipopoly- saccharide and concanavalin A was elevated on duty 7 after infection the mice transinoculated with immune splenocytes. The serum antibody titers in the mice transferred with immune spleno- cytes were increased gradually from day 7 up to day 20 after infections by mean of ELISA. It is suggested that the transfer of splenocytes from immuniged mice conferred immunity against N. fowleri infection.
Atopic dermatitis is a chronic inflammatory skin disease caused by a variety of genetic and environmental factors, dysregulation of immunological response, as well as dysfunction of the skin barrier proteins. The purpose of this study is to develop an ELISA kit suitable for evaluating the expression of skin barrier proteins. Proteins were obtained from the skin via AriNo and D-Squame patches. The efficiency of protein collection from the skin, using the Arino patch, was shown to be more effective than using D-Squame; while the efficiency of lysis using 0.1% Triton-X100 was higher than that of other lysis solutions, including 0.1 M Tris-HCL, 0.1% Tween-20, and 5 mM KOH. Recombinant skin barrier proteins, such as filaggrin and involucrin, were produced by molecular biological methods. Monoclonal antibodies against filaggrin and involucrin were produced by immunization of mice, fusion of spleen cells and myeloma cells, as well as a selection of antibody-producing hybridoma cells. The filaggrin expression in the skin of subjects suffering from atopic dermatitis was lower than that in normal mice. Involucrin expression was not altered between normal individuals and subjects with atopic dermatitis. These findings contribute to an elucidation of the importance of the skin barrier protein expression in atopic dermatitis and the development of a diagnostic kit for atopic dermatitis.
Proceedings of the Microbiological Society of Korea Conference
/
2008.05a
/
pp.23-25
/
2008
Serum complement proteins comprise an important system that is responsible for several innate and adaptive immune defence mechanisms. There were three well described pathways known to lead to the generation of a C3 convertase, which catalyses the proteolysis of complement component C3, and leads to the formation of C3 opsonins (C3b, iC3b and C3d) that fix to bacteria. A pivotal step in the complement pathway is the assembly of a C3 convertase, which digests the C3 complement component to form microbial-binding C3 fragments recognized by leukocytes. The spleen clears microorganisms from the blood. Individuals lacking this organ are more susceptible to Streptococcus pneumoniae. Innate resistance to S. pneumoniae has previously been shown to involve complement components C3 and C4, however this resistance has only a partial requirement for mediators of these three pathways, such as immunoglobulin, factor B and mannose-binding lectin. Therefore it was likely that spleen and complement system provide resistance against blood-borne S. pneumoniae infection through unknown mechanism. To better understand the mechanisms involved, we studied Specific intracellular adhesion molecule-grabbing nonintegrin (SIGN)-R1. SIGN-R1, is a C-type lectin that is expressed at high levels by spleen marginal-zone macrophages and lymph-node macrophages. SIGN-R1 has previously been shown to be the main receptor for bacterial dextrans, as well as for the capsular pneumococcal polysaccharide (CPS) of S. pneumoniae. We examined the specific role of this receptor in the activation of complement. Using a monoclonal antibody that selectively downregulates SIGN-R1 expression in vivo, we show that in response to S. pneumoniae or CPS, SIGN-R1 mediates the immediate proteolysis of C3 and fixation of C3 opsonins to S. pneumoniae or to marginal-zone macrophages that had taken up CPS. These data indicate that SIGN-R1 is largely responsible for the rapid C3 convertase formation induced by S. pneumoniae in the spleen of mice. Also, we found that SIGN-R1 directly binds C1q and that C3 fixation by SIGN-R1 requires C1q and C4 but not factor B or immunoglobulin. Traditionally C3 convertase can be formed by the classical C1q- and immunoglobulin-dependent pathway, the alternative factor-B-dependent pathway and the soluble mannose-binding lectin pathway. Furthermore Conditional SIGN-R1 knockout mice developed deficits in C3 catabolism when given S. pneumoniae or its capsular polysaccharide intravenously. There were marked reductions in proteolysis of serum C3, deposition of C3 on organisms within SIGN-$R1^+$ spleen macrophages, and formation of C3 ligands. The transmembrane lectin SIGN-R1 therefore contributes to innate resistance by an unusual C3 activation pathway. We propose that in the SIGN-R1 mediated complement activation pathway, after binding to polysaccharide, SIGN-R1 captures C1q. SIGN-R1 can then, in association with several other complement proteins including C4, lead to the formation of a C3 convertase and fixation of C3. Therefore, this new pathway for C3 fixation by SIGN-R1, which is unusual as it is a classical C1q-dependent pathway that does not require immuno globulin, contributes to innate immune resistance to certain encapsulated microorganisms.
Fayoumi chicks were vaccinated with fowl pox virus vaccine and pigeon pox virus vaccine. The protective potentiality of the two vaccines was compared in field condition in Bangladesh. The percentage of 'take reaction' was assessed to conclude its relationship with better immune response and recorded 93.33% and 100% in birds of group B and group C, respectively. The mean passive hemagglutination (PHA) antibody titre after primary vaccination was $32{\pm}14.81$ in group B and $33{\pm}13.66$ in group C. Following booster vaccination, the mean PHA titres level at pre challenge of group B was $46.93{\pm}16.52\;and\;55.46{\pm}14.64$ in group C. The PHA titre of group B and C at two weeks post challenge recorded $93.86{\pm}33.04\;and\;110.93{\pm}29.29$, respectively. PHA titre significantly (P<0.01) increased after vaccination and post challenge compared to pre- vaccination titre. There was significant variation (p<0.01) of PHA titre at pre challenge in these groups using different vaccine combinations, but all the vaccinated birds resisted challenge infection.
Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.
Shin, Donghyun;Kim, Kyoung Hwan;Lee, Ji Hyun;Cho, Byung-Wook
Journal of Life Science
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v.28
no.10
/
pp.1127-1131
/
2018
Cortisol, a steroid hormone, functions within metabolism, immune response, and stress. Intense or prolonged physical exercise increases cortisol levels to enhance the gluconeogenesis pathway and stabilize blood glucose level. However, cortisol also exerts a negative impact on muscle function and creates a stressful environment in skeletal muscle cells. The present study investigated the function of cortisol as a stress hormone. To examine the effect of the exercise-induced hormone cortisol on skeletal muscles, C2C12 cells were cultured and treated with cortisol at different concentrations. As a result, we found that the morphology of C2C12 changed remarkably with 5 ug/ml cortisol treatment. Western blot analysis was conducted to learn whether ER-stress and autophagy were induced. We found that the expression ratio of LC3I/LC3II decreased and BiP expression increased after cortisol treatment. In addition, immunocytochemistry analysis with IER3 antibody clearly showed that apoptosis is induced after 12-hour cortisol treatment. These results indicate that cortisol treatment could induce apoptosis, ER-stress, and autophagy in muscle cells. This study would provide valuable information in the study of the effects of exercise on skeletal muscle cells and the development of additives to reduce cortisol stress.
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