The goal of this study was to examine whether lipopolysaccharide (LPS) induces the expression of proinflammatory cytokines and chemokines and recruits T cells in the lower part of the oviduct, and whether that response to LPS is different between the laying and molting phase. White Leghorn laying and molting hens were intravenously injected with saline (control) or LPS. The uterus and vagina of oviducts were collected 3 or 6 h after injection, and used for reverse transcription PCR analysis of IL-1β, IL-6, IL-8 (CXCLi2), and lymphotactin (Lptn), and for immunohistochemical analysis for the frequency of CD4+ and CD8+ T cells. The expressions of IL-1β, IL-6, and CXCLi2 in the uterus and that of IL-1β in the vagina were upregulated in response to LPS 3 or 6 h after injection in both laying and molting hens. The CXCLi2 expression in the vagina was upregulated by LPS in laying hens, whereas those effects of LPS were not significant in molting hens. Expression of Lptn showed a tendency to be downregulated after 3 h, with recovery by 6 h after LPS injection. The frequency of CD4+ T cells tended to increase in response to LPS after 6 h in the lamina propria of the uterus and vagina in both laying and molting hens. The CD8+ T cell frequencies in the lamina propria of the uterus and vagina of laying hens increased in response to LPS after 6 h. However, in the molting hens, LPS stimulation resulted in CD8+ T cell increase in the vagina only and not in the uterus. These results suggest that expressions of proinflammatory cytokines and CXCLi2 chemokine are upregulated in association with T cell recruitment in response to LPS in the lower part of the oviduct, although CD8+ T cells in the uterus may be depressed during the molting phase. These immunoresponses may play roles in the defense against infection of the oviduct.

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[reaction: see text] A ruthenium catalyst precursor bearing a bulky and electron-donating pentamethylcyclopentadienyl (Cp) ligand, typically CpRuH3(PPh3), mediates hydrosilylation of several 1-alkynes with novel regioselectivity to give preferentially 2-silyl-1-alkenes.

Immune function in the vagina of hen oviduct is essential to prevent infection by microorganisms colonizing in the cloaca. The aim of this study was to determine whether CpG oligodeoxynucleotides (CpG-ODN) stimulate the expression of avian b-defensins (AvBDs) in hen vaginal cells. Specific questions were whether CpG-ODN affects the expression of AvBDs and proinflammatory cytokines and whether the cytokines affect AvBDs expression in vaginal cells. The dispersed vaginal cells of White Leghorn laying hens were cultured and stimulated by different doses of lipopolysaccharide (LPS), CpG-ODN, interleukin 1b (IL1B), or IL6. The cultured cell population contained epithelial cells, fibroblast-like cells, and CD45-positive leukocytes. The immunoreactive AvBD3, -10, and -12 were localized in the mucosal epithelium in the section of the vagina. The expression of AvBDs, IL1B, and IL6 was analyzed by quantitative RT-PCR. RT-PCR analysis showed the expression of AvBD1, -3, -4, -5, -10, and -12 in the cultured vaginal cells without stimulation. Toll-like receptors (TLRs) 4 and 21, which recognize LPS and CpG-ODN respectively and IL1 and IL6 receptors (IL1R1 and IL6R) were also expressed in them. The expression of IL1B, IL6, and AvBD10 and -12 was upregulated by LPS, whereas only IL1B and IL6 were upregulated by CpG-ODN. IL1B stimulation upregulated AvBD1 and -3 expression, whereas IL6 stimulation did not cause changes in AvBDs expression. These results suggest that CpG-ODN derived from microbes upregulates the expression of IL1B and IL6 by interaction with TLR21 and then IL1B induces AvBD1 and -3 to prevent infection in the vagina.

Hepatic stellate cells (HSCs) are key players in liver fibrosis and regeneration via collagen degradation and synthesis. These phenomena involve inflammatory cytokines released from non-parenchymal liver cells such as Kupffer cells. Although the effects of individual cytokines on many cell types have been investigated in various conditions, such as inflammation and tissue fibrosis, investigating the effect of combined cytokines would further our understanding of the regulatory mechanisms in tissue fibrosis. Here, we report the effect of multiple cytokine combinations on primary HSCs. We first examined the effect of individual cytokines and then the simultaneous exposure of different cytokines, including interleukin-6 (IL-6), IL-1 alpha (IL-1α), platelet-derived growth factor (PDGF), tumour necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β), on matrix metalloproteinase-1 (MMP1) gene expression in primary HSCs. We observed that the combination of all five cytokines induced higher levels of MMP1 gene expression. Of these cytokines, TNF-α and IL-1α were found to be the key cytokines for not only inducing MMP1 expression, but also increasing α-smooth muscle actin gene expression. In conclusion, the combined treatment of TNF-α and IL-1α on HSCs had an enhanced effect on the expression of the fibrotic genes, MMP1 and α-smooth muscle actin, so appears to be an important regulator for tissue regeneration. This finding suggests that stimulation with combined anti-fibrotic cytokines is a potential approach in the development of a novel therapy for the recovery of liver fibrosis.

Palladium complex (A)-catalyzed dimerization-hydrosilylation of 1-alkynes, which has previously been reported, can not be applied effectively to the intramolecular version of a,walkanediynes, while a cationic palladium complex (B) is a very effective catalyst for cyclization-hydrosilylation of functionalized a,w-diynes to give (Z)-1-methylene-2-silylmethylenecycloalkane derivatives, with 5-, 6-, and 7-membered ring, in good to moderate yields.

In a previous paper," it has been reported that several ninhydrinpositive ampholytes, which are stable to acid hydrolysis and give the unusual large spots on paper chromatogram, are found to be excreted in urine of a hyperthyroid patient. Among the ampholytes mentioned above sarcosine and ,9-alanine have already been identified on paper chromatogram by using a known sample of each. Subsequent study 2' has revealed that thyroxine treatment has led to the increased excretion of lysine, sarcosine, ,9-alanine, and an unidentified ampholyte in rabbit urine. This communication is concerned with further identification of three unidentified amino acids excreted by a hyperthyroid patient. Now it seems quite probable that these amino acids are pipecolic acid,-aminovaleric acid, and (?-aminoisobutyric acid. Experimental. Methods. As described previously,3' urine desalted by the method of Carsten 4' was applied on a filter paper, and n-butanolacetic acid-water (5 :1.2 : 5 v/v) and phenol-water (4: 1 v/v, 0.3°o NH3 in the chamber) were used as the solvents for two-way chromatography. O.1% ninhydrin in water-saturated n-butanol was used for color development. Materials. /3-Aminoisobutyric acid, a-aminobutyric acid, and ryaminobutyric acid were kindly furnished by Prof. Dr. K. Ichihara, and pipecolic acid was a generous gift from Prof. Dr. M. Suda. ~S-Aminovaleric acid was synthesized by a modification of Wallach's method.5' The other amino acids tested were commercial products. Results. Fig. 1 shows the urinary amino acid pattern of a hyperthyroid patient. Spots (a) and (b) were previously identified to be Salanine and sarcosine respectively.'' Urinary serine is usually contained in fairly large amounts. But in this patient, serine almost completely disappeared from the urine while sarcosine increased. This fact has agreed with the observation that sarcosine oxidase activity and serine formation in rat liver are reduced when the rat is in a hyperthyroid state.6' When f3-aminoisobutyric acid and pipecolic acid were spotted on a zero point with the urine sample of the patient, these amino acids have just overlapped on the spot (c) and (e) respectively as shown in

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