Expression of CD4 T cell cytokine genes in the colorectal mucosa of inflammatory colorectal polyps in miniature dachshunds

Inflammatory colorectal polyps (ICRPs) in miniature dachshunds are recently recognized as a major cause of large bowel diarrhea in this dog breed in Japan. ICRPs are characterized by the formation of multiple small polyps and a space-occupying large polyp in the colorectal area, and are thought to be a novel form of inflammatory bowel disease (IBD). In humans, specific cytokine patterns attributed to T helper (Th)1, Th17 and regulatory T cells have important roles in the pathogenesis of IBD. Thus, the aim of the present study was to assess the gene expression of cytokines of T cell subsets in the colorectal mucosa from dogs with ICRPs. Colorectal mucosal specimens from 10 dogs with ICRPs and 14 control dogs were used in this study. Interferon (IFN)-γ, interleukin (IL)-4, IL-17A and IL-10 mRNA expression was assessed using quantitative real-time PCR. IL-17A mRNA expression was significantly increased in large polyps compared to small polyps and controls. IFN-γ and IL-10 mRNA expression in large polyps were significantly higher than in controls. There was no significant difference in IL-4 mRNA expression among the three groups. IL-17A is thought to play important roles in the pathogenesis of ICRPs. IL-10 up-regulation could oppose the proinflammatory function of IL-17A.     

Efficacy of thiabendazole, mebendazole, levamisole and ivermectin against gullet worm, Gongylonema pulchrum: in vitro and in vivo studies

In vitro and in vivo studies were conducted to evaluate the effects of thiabendazole, mebendazole, levamisole and ivermectin against Gongylonema pulchrum. For in vitro assays, third-stage larvae (L3) incubated with the drugs were administered orally to mice and the ability of larvae to invade the gastric mucosa of the animals was examined. After incubation, only those larvae treated with high concentrations of levamisole (1 and 10 microg/ml) were tightly coiled with intestines exhibiting morphological abnormalities. Good dose-response data for the drugs tested was observed at the time of worm recovery from mice, with no worms recovered at the two highest concentrations of levamisole. In vivo efficacy of the drugs against adult worms was evaluated in six groups of three rabbits, each of which was infected with 30 L3 of G. pulchrum and treated with thiabendazole at 100 mg/kg for 3 days, mebendazole at 70 mg/kg for 3 days, levamisole as a single dose of 8 mg/kg, and subcutaneously injected ivermectin as a single dose of 0.2 mg/kg or vehicles of the drugs (control) at 4 months post-infection. Necropsy 14 days after treatment revealed that levamisole, mebendazole and ivermectin reduced worm burdens by 63.2%, 22.8% and 25.8%, respectively, with no reductions in worms observed with thiabendazole. The surviving worms were principally found in the esophagus with the remainder distributed among the buccal mucosa, the tongue, and/or pharyngeal mucosa in all groups. A number of morphologically abnormal eggs were observed within the uterus and ovijector in female worms recovered from the thiabendazole-treated group. These findings suggest that levamisole exhibits in vivo efficacy against G. pulchrum infection and that the larval invasion tests using mice could be used to screen for anthelmintic susceptibility of nematodes.     

Effect of CD4+CD25+ T cell-depletion on acute lethal infection of mice with Trypanosoma congolense

Despite the immense socio-economic repercussions of African trypanosomosis (AT), there is currently no effective control measure against the disease. Characterization of mechanisms governing resistance and/or susceptibility to AT could suggest interventions that might lead to more effective disease control. The present study was designed in an attempt to address the possible role of CD4+CD25+ T cells during an acute lethal infection of mice with Trypanosoma congolense, the causative agent of AT in domestic animals, through selective depletion using anti-CD25 monoclonal antibody. Accordingly, CD4+CD25+ T-cell-depletion resulted in a significant reduction or delay in parasitemia, pathology, and mortality, as compared to controls. The apparent resistance in CD4+CD25+-T-cell-depleted mice correlated with a profound suppression of Th2 cytokines in vitro and in vivo, culminating in a net Th1 cytokine environment. Cumulatively, these findings suggest that CD4+CD25+ T-cell- depletion improves the trypanotolerance of highly susceptible BALB/c mice acutely infected with the lethal T. congolense.     

cFLIP regulates death receptor-mediated apoptosis in an ovarian granulosa cell line by inhibiting procaspase-8 cleavage

More than 99% of follicles in mammalian ovaries undergo atresia, but the mechanisms regulating the strict selection process are still unclear. Granulosa cell apoptosis is considered the trigger of follicular atresia, which occurs in advance of the death of an oocyte. Cellular FLICE-like inhibitory protein (cFLIP), a homologue of procaspase-8 (also called FLICE), is an intracellular anti-apoptotic protein. It is expressed in granulosa cells of porcine ovaries, where its levels decreases during follicular atresia. We hypothesized that cFLIP regulates granulosa cell apoptosis by acting as a pro-survival factor. In the present study, to further reveal the function of cFLIP in granulosa cells, we examined the anti-apoptotic mechanism of cFLIP using KGN, a human granulosa tumor cell line. Fas-mediated apoptosis was induced by co-treatment with anti-Fas antibody (CH-11), which acts as an agonist of Fas-ligand, and cycloheximide (CHX). When cFLIP was stably expressed in KGN cells following transfection of an expression vector, the Fas-mediated apoptosis was inhibited. Suppression of cFLIP by small interfering RNA (siRNA) spontaneously induced cell death. Silencing of cFLIP promoted cleavage of procaspase-8, and the cell death caused by cFLIP siRNA was completely blocked by a caspase-8 inhibitor (Z-IETD-FMK), indicating that cFLIP regulates apoptosis in KGN cells by inhibiting cleavage of procaspase-8. In conclusion, cFLIP is an essential pro-survival factor for granulosa cells, and it prevents granulosa cell apoptosis by inhibiting procaspase-8 activation.     

Growth and age composition of northern shrimp Pandalus eous estimated by multiple length frequency analysis

We examined growth of northern shrimp Pandalus eous in the Sea of Japan, off western Hokkaido, to improve estimations of catch-at-age for stock assessment. Multiple length frequency analysis based on length frequency data collected by a scientific research vessel was conducted to examine length-at-age in the shrimp population. Multi-normal distributions estimated using maximum likelihood indicated a good fit to length distributions. AIC values and regression analyses revealed annual growth variation and a decreasing trend in the length at several age classes in the shrimp population. We revised the method for estimating catch-at-age from the age-conversion table (ACT), which is a simple method for age determination, to age–length keys (ALK) calculated from the results of multiple length frequency analysis. Abundant year classes caught successively year after year could be more easily identified from the catch-at-age data computed using ALK than by using ACT. Our results suggested not only that the mean size of commercial landings fluctuated based on changes in age composition but also that a decrease in the length-at-age in the population influenced the consistent size decrease of commercial landings.     

Energy partitioning in cultured juvenile Pacific bluefin tuna,Thunnus orientalis (Temminck & Schlegel, 1844)

The characteristics of dietary utilization, energy conversion efficiency, metabolic rate and energy partitioning were measured for cultured Pacific bluefin tuna (PBT) juveniles fed on an artificial diet. Thirty‐one juveniles (1.1 ± 0.3 g BW) were stocked into each of two 2500 L tanks to measure oxygen consumption (?O₂), swimming speed, digestibility and growth performance. ?O₂ elevated until 2.5 ± 0.3 times of pre‐feeding level within 1.5 h after feeding, except for the first feeding, and returned to pre‐feeding level within 3 h. Swimming speed fluctuation was corresponded with the ?O₂ fluctuation, and both parameters were stable from 02:00 to 06:00 and also during the whole day for starved fish. These indicate that feeding has strong influence on their metabolic rate. Energy partitioning for faecal, urinary and branchial, heat increment and voluntary activity, standard metabolism, and retained energy were calculated to be 17.2%, 5.9%, 14.9%, 41.3% and 20.7% of total ingested energy, respectively. The results indicate that, unlike other fish, juvenile PBT distribute large amount of energy for maintenance, which allows only a little proportion of ingested energy available for growth.