The immune modifying effects of amino acids on gut-associated lymphoid tissue

The intestine and the gut-associated lymphoid tissue（GALT） are essential components of whole body immune defense,protecting the body from foreign antigens and pathogens,while allowing tolerance to commensal bacteria and dietary antigens.The requirement for protein to support the immune system is well established.Less is known regarding the immune modifying properties of individual amino acids,particularly on the GALT.Both oral and parenteral feeding studies have established convincing evidence that not only the total protein intake,but the availability of specific dietary amino acids（in particular glutamine,glutamate,and arginine,and perhaps methionine,cysteine and threonine） are essential to optimizing the immune functions of the intestine and the proximal resident immune cells.These amino acids each have unique properties that include,maintaining the integrity,growth and function of the intestine,as well as normalizing inflammatory cytokine secretion and improving T-lymphocyte numbers,specific T cell functions,and the secretion of IgA by lamina propria cells.Our understanding of this area has come from studies that have supplemented single amino acids to a mixed protein diet and measuring the effect on specific immune parameters.Future studies should be designed using amino acid mixtures that target a number of specific functions of GALT in order to optimize immune function in domestic animals and humans during critical periods of development and various disease states.     

莱芜黑猪a1岩藻糖转移酶基因(FUT I)不同基因型对大肠杆菌F18抵抗力研究

为了研究不同猪种(山东省地方猪种莱芜黑猪和引进猪种大约克)和FUT I不同基因型仔猪对肠毒性大肠杆菌F18(ETEC F18)的抵抗能力和肠黏膜上皮细胞对ETEC F18 黏附能力的差异,在采用PCR-RFLP技术对莱芜黑猪和大约克FUT I基因型检测的基础上,选取易感性和抗性断奶仔猪,利用野生型ETEC F18标准株进行试验猪口服细菌攻毒试验和肠黏膜黏附试验.由于多方面原因,接受ETEC F18细菌攻毒的试验猪均未发病,无法判断试验猪对ETEC F18的抵抗能力.ETEC F18标准菌株能与所有FUT I敏感基因型仔猪的小肠黏膜上皮细胞黏附,而不能与抗性基因型仔猪小肠上皮黏膜细胞黏附,莱芜黑猪与大约克的肠黏膜上皮细胞与E.coli F18的黏附特性并无品种差异.但是养猪生产实践中,莱芜黑猪极少发生断奶仔猪水肿和腹泻病.所以除FUT I基因外,也许本地猪种有其他导致遗传抗性的突变或抗性基因.     

Role of PARP and caspase-3 in the airway epithelial injury induced by peroxynitrite

AIM:To study the mechanism responsible for ONOO^--induced the airway epithelial injury. METHODS:Effects of 3-aminobenzamide(3-AB), a poly-(ADP-ribose) polymerase(PARP) inhibitor, and Ac-DEVD-CHO, a caspase-3 inhibitor, on LDH release and apoptosis of cultured rat tracheal epithelial (RTE) cells induced by ONOO^- were examined. The cleavage of PARP was analysed by Western blot. RESULTS:3-AB inhibited the release of LDH induced by ONOO^- partially, and had no effect on the apoptosis of RTE cells. Caspase-3 inhibitor Ac-DEVD-CHO obviously prevented the apoptosis of RTE cells induced by ONOO^- in a dose-dependent manner. The cleavage of PARP was observed in the process of apoptosis of RTE cells induced by ONOO^-. CONCLUSIONS:PARP activation represents one of the pathways of ONOO^--mediated epithelial injury, and the excessive activation of PARP contributes to the necrosis in RTE cells induced by ONOO^-. Cleavage of PARP by activated caspase-3 plays a crucial role in the apoptosis of RTE cells induced by ONOO^-.     

Effects of electrolyzed free radicals on endothelium of isolated guinea pig coronary and epithelium of airway tube

AIM and METHODS:To study the damage effects of free radicals from electrolyzed krebs solution(direct current,10 mA,1-2 min) on isolated guinea pig coronary and airway tube. RESULTS:In Langendorff’s perfused guinea pig hearts,the electrolyzed free radicals increased coronary perfusion pressure(4.4±1.2) kPa,inhibited myocyte contractility [(0.8±0.8) g vs (2 9±0 6) g, P< 0.05],increased TBARS level and decreased SOD activity.In isolated perfused lungs of guinia pig,electrolyzed Krebs solution promoted significantly the airway perfusion pressure [(0.03±0.01) kPa vs (2.20±0.29) kPa, P< 0.01] and histamine reactivity [(0.65±0.37) kPa vs (2.05±0.25) kPa, P< 0.01]. Hydroxyl radicals scavenger DMSO and natural medicine gypenosides prevented the effects of oxygen free radicals. CONCLUSION: These results indicated that the free radicals by electrolyzation could induce damages of coronary endothelium and airway epithelium.     

Polarized epithelium-sperm co-culture system reveals stimulatory factors for the secretion of mouse epididymal quiescin sulfhydryl oxidase 1

Spermatozoa acquire fertilization ability through post-translational modifications. These membrane surface alterations occur in various segments of the epididymis. Quiescin sulfhydryl oxidases, which catalyze thiol-oxidation reactions, are involved in disulfide bond formation, which is essential for sperm maturation, upon transition and migration in the epididymis. Using castration and azoospermia transgenic mouse models, in the present study, we showed that quiescin sulfhydryl oxidase 1 (QSOX1) protein expression and secretion are positively correlated with the presence of testosterone and sperm cells. A two-dimensional in vitro epithelium-sperm co-culture system provided further evidence in support of the notion that both testosterone and its dominant metabolite, 5α-dihydrotestosterone, promote epididymal QSOX1 secretion. We also demonstrated that immature caput spermatozoa, but not mature cauda sperm cells, exhibited great potential to stimulate QSOX1 secretion in vitro, suggesting that sperm maturation is a key regulatory factor for mouse epididymal QSOX1 secretion. Proteomic analysis identified 582 secretory proteins from the co-culture supernatant, of which 258 were sperm-specific and 154 were of epididymal epithelium-origin. Gene Ontology analysis indicated that these secreted proteins exhibit functions known to facilitate sperm membrane organization, cellular activity, and sperm-egg recognition. Taken together, our data demonstrated that testosterone and sperm maturation status are key regulators of mouse epididymal QSOX1 protein expression and secretion.     

The immune modifying effects of amino acids on gut-associated lymphoid tissue

The intestine and the gut-associated lymphoid tissue (GALT) are essential components of whole body immune defense, protecting the body from foreign antigens and pathogens, while allowing tolerance to commensal bacteria and dietary antigens. The requirement for protein to support the immune system is well established. Less is known regarding the immune modifying properties of individual amino acids, particularly on the GALT. Both oral and parenteral feeding studies have established convincing evidence that not only the total protein intake, but the availability of specific dietary amino acids (in particular glutamine, glutamate, and arginine, and perhaps methionine, cysteine and threonine) are essential to optimizing the immune functions of the intestine and the proximal resident immune cells. These amino acids each have unique properties that include, maintaining the integrity, growth and function of the intestine, as well as normalizing inflammatory cytokine secretion and improving T-lymphocyte numbers, specific T cell functions, and the secretion of IgA by lamina propria cells. Our understanding of this area has come from studies that have supplemented single amino acids to a mixed protein diet and measuring the effect on specific immune parameters. Future studies should be designed using amino acid mixtures that target a number of specific functions of GALT in order to optimize immune function in domestic animals and humans during critical periods of development and various disease states.     

Relationship of equine housing to large airway inflammation

The pasture, as compared to stall confinement, has been considered a more desirable environment for maintaining the health of the respiratory system in the horse. This conclusion is based on reports which showed that ventilation in most barns was generally poor, and that horses bedded on straw and fed hay were exposed to many forms of respirable debris. In this study, six normal horses were evaluated for evidence of airway mucosal inflammation after being housed on pasture or stabled in a barn for one month. The response of the horses' airways was measured by assigning scores for the degree of tracheal mucosal secretions that were observed by endoscopic visualization. Cytological examination of transtracheal wash secretions was also performed, as well as histologic evaluation of tracheobronchial tissues obtained by a transendoscopic epithelial biopsy technique. Samples were collected at three time points; the initial collection occurred after the horses were housed on pasture for one month. The horses were subsequently moved to a barn for an equal length of time and samples were obtained at the end of this period. The horses were then returned to their original pasture and final sampling was performed after they were housed in this environment for two months. There were no significant changes in any of the parameters evaluated, regardless of the environment in which the horses were maintained. These findings indicate that housing horses in a barn for four weeks does not cause tracheobronchial mucosal inflammation in a manner that could be detected using the methods employed in this study.     

黄牛输卵管上皮细胞的体外分离与培养

为探讨适用于黄牛输卵管单层上皮细胞的培养方法，采用机械剪取及0．25％胰酶消化的方法分离获得上皮细胞，取对数生长期细胞进行MTT比色实验检测细胞活力和生长速度；台盼蓝排斥试验检测活细胞数。结果表明利用机械剪取及0．25％胰酶消化的方法经细胞形态观察可以分离获得黄牛输卵管上皮单层细胞；台盼蓝排斥实验经计数计算传代的上皮细胞活力在90％以上；经冷冻后再解冻的输卵管上皮细胞活力在75％以上。该研究探索建立了一套较为适宜的黄牛输卵管上皮细胞分离、培养模型。     

Canine normal corneal epithelium bears a large population of CD45-positive cells

This investigation sought to identify the presence of immune cells in normal canine corneal epithelium. A whole-mount immunofluorescence study of normal canine epithelium using monoclonal antibodies against CD45, CD11c, CD1c and MHC class II was performed. CD45-positive cells were located in all epithelial layers throughout the cornea, occurring in greater numbers (51.98 ± 4.1/mm²) at the periphery and decreasing towards the central region (11.8 ± 3.1/mm²). CD11c-positive cells were also observed, but were fewer in number. The findings show that the normal canine cornea carries a significant number of cells of immune origin; these cells seem to be of an inactive phenotype as they do not express MHC class II. Further studies are needed to determine whether these cells can express co-stimulatory molecules and act as antigen presenting cells if stimulated.