In this study, we investigated the possibility that GLUT4 protein concentration and mitochondrial enzyme activity is coregulated by muscle contractile activity. In the first experiment, male rats were trained by treadmill running or swimming for 3 wks and training effects in upper-and hind-limb muscle were investigated. Treadmill training program induced increases of 30% in citrate synthase activity, 51% in hexokinase activity, and 35% in GLUT4 protein concentration in hind-limb soleus muscles without causing training effects in upper-limb epitrochlearis muscles. On the other hand, swimming training program induced increases of 67% in citrate synthase activity, 139% in hexokinase activity, and 89% in GLUT4 protein concentration in epitrochlearis muscle without causing training effects in soleus muscles. These results suggest that muscle contractile activity which raises mitochondrial enzyme activity increases GLUT4 protein concentration in skeletal muscle. In a second set of experiment, we investigated relationship between muscle GLUT4 protein concentration and mitochondrial enzyme activity by use of the various type of rat muscles (soleus, plantaris, red gastrocnemius, white gastrocnemius, epitrochlearis and heart), which possese different contractile activity level. Significant correlation was observed between GLUT4 protein concentration and citrate synthase activity among different muscles (r=0.936) . They suggest that muscle GLUT4 protein concentration and mitochondrial enzyme activity level may be coregulated according to muscle contractile activity level.

A simplified polymerase chain reaction (PCR) assay was developed for fast and easy screening of mycoplasma mastitis in dairy cattle. Species of major mycoplasma strains [Mycoplasma (M.) bovis, M. arginini, M. bovigenitalium, M. californicum, M. bovirhinis, M. alkalescens and M. canadense] in cultured milk samples were detected by this simplified PCR-based method as well as a standard PCR technique. The minimum concentration limit for detecting mycoplasma by the simplified PCR was estimated to be about 2.5 x 10(3) cfu/mL and was similar to that of the standard PCR. We compared the specificity and sensitivity of the simplified PCR to those of a culture method. Out of 1,685 milk samples cultured in mycoplasma broth, the simplified PCR detected Mycoplasma DNA in 152 that were also positive according to the culture assay. The sensitivity and specificity of the simplified PCR were 98.7% and 99.7%, respectively, for detecting mycoplasma in those cultures. The results obtained by the simplified PCR were consistent with ones from standard PCR. This newly developed simplified PCR, which does not require DNA purification, can analyze about 300 cultured samples within 3 h. The results from our study suggest that the simplified PCR can be used for mycoplasma mastitis screening in large-scale dairy farms.
Animals ; Cattle ; Colony Count, Microbial/veterinary ; DNA, Bacterial/chemistry/genetics ; Disease Outbreaks/prevention & control/veterinary ; Female ; Mastitis, Bovine/diagnosis/*microbiology ; Milk/cytology/*microbiology ; Mycoplasma/genetics/*isolation & purification ; Mycoplasma Infections/diagnosis/microbiology/*veterinary ; Polymerase Chain Reaction/veterinary

Mycoplasma spp. are contagious bacteria, and mycoplasmal mastitis is a serious productivity problem on dairy farms. Bovine mammary epithelial cells (bMECs) have an important role in the elimination of pathogens, but the effect of Mycoplasma bovis on bMECs has not been fully described. To elucidate the immune response against intramammary infection by M. bovis, we undertook microarray analysis to examine and profile mRNA expression in bMECs after stimulation with M. bovis. We also compared the effects of M. bovis, Staphylococcus aureus, and Escherichia coli on immune-related mRNA expression in bMECs. Transcriptome analysis indicated a significant decrease in the level of mRNA-encoding lysine-specific demethylase 4D, suggesting that the immune response is suppressed by a decrease in histone demethylase activity. Interleukin (IL)-1β, IL-6, tumor necrosis factor alpha, toll-like receptor (TLR) 2, and TLR4 mRNA expression levels were significantly increased in bMECs stimulated with heat-killed M. bovis, but the expression levels were lower than those following stimulation by heat-killed S. aureus or E. coli. Our results suggest that M. bovis weakly affects mRNA expression in bMECs compared to the effects of E. coli or S. aureus. Moreover, live M. bovis may induce suppression of the immune response in bMECs.
Agriculture ; Bacteria ; Cytokines ; Efficiency ; Epithelial Cells ; Escherichia coli ; Female ; Gene Expression Profiling ; Histones ; Immunity, Innate ; Interleukin-6 ; Interleukins ; Mastitis ; Microarray Analysis ; Mycoplasma bovis ; Mycoplasma ; RNA, Messenger ; Staphylococcus aureus ; Toll-Like Receptors ; Tumor Necrosis Factor-alpha

We report herein improved methods for the safe and successful completion of endoscopic papillectomy (EP). Between January 2008 and November 2011, 12 patients underwent double-snare retracting papillectomy for the treatment of lesions of the major duodenal papilla. The main outcomes were en bloc resection rates, pathological findings, and adverse events. All of the patients (mean age, 60.1 years; range, 38 to 80 years) were diagnosed with ampullary adenoma by endoscopic forceps biopsies prior to endoscopic snare papillectomy. En bloc resection by double-snare retracting papillectomy was successfully performed for all lesions (median size, 12.3 mm), comprising six tubular adenomas, one tubulovillous adenoma, three cases of epithelial atypia, one hamartomatous polyp, and one case of duodenitis with regenerative change. Significant hemorrhage and pancreatitis were observed in one case after EP. Adenoma recurrence occurred in three patients during follow-up (median, 28.5 months) at a mean interval of 2 months postoperatively (range, 1 to 3 months). No serious adverse events were observed. Double-snare retracting papillectomy is effective and feasible for treating lesions of the major duodenal papilla. Further treatment experience, including a single-arm phase II study, needs to be accumulated before conducting a randomized controlled study.
Adenoma/pathology/*surgery ; Adult ; Aged ; Aged, 80 and over ; Ampulla of Vater/pathology/*surgery ; Biopsy ; Common Bile Duct Neoplasms/pathology/*surgery ; Dissection/*methods ; Duodenoscopy/*methods ; Feasibility Studies ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Treatment Outcome