The receptor for advanced glycation end-products (RAGE)-mediated cellular activation through the mitogen-activated protein kinase (MAPK) cascade, activation of NF-U UB and Rho family small G-proteins, cdc42/Rac, is implicated in the pathogenesis of in£ammatory disorders and tumor growth/metastasis. However, the precise molecular mechanisms for the initiation of cell signaling by RAGE remain to be elucidated. In this study, proteins which directly bind to the cytoplasmic C-terminus of RAGE were puri¢ed from rat lung extracts using an a⁄nity chromatography technique and identi¢ed to be extracellular signal-regulated protein kinase-1 and -2 (ERK-1/2). Their interactions were con¢rmed by immunoprecipitation of ERK-1/2 from RAGE-expressing HT1080 cell extracts with anti-RAGE antibody. Furthermore, the augmentation of kinase activity of RAGE-bound ERK upon the stimulation of cells with amphoterin was demonstrated by determining the phosphorylation level of myelin basic protein, an ERK substrate. In vitro binding studies using a series of C-terminal deletion mutants of human RAGE revealed the importance of the membrane-proximal cytoplasmic region of RAGE for the direct ERK^RAGE interaction. This region contained a sequence similar to the D-domain, a ERK docking site which is conserved in some ERK substrates including MAPK-interacting kinase-1/2, mitogen-and stress-activated protein kinase-1, and ribosomal S6 kinase. These data suggest that ERK may play a role in RAGE signaling through direct interaction with RAGE. ß
A nationwide surveillance for antimicrobial susceptibility in Escherichia coli strains isolated from foodproducing animals in Japan was conducted from 1999 to 2002. Eighteen cefazolin-resistant E. coli strains were isolated from broilers. Six were CTX-M-type producing, and eight were CMY-2 producing, while eight had mutations at the ampC promoter region.
Recently, methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP) have been increasingly isolated from veterinarians and companion animals. With a view to preventing the spread of MRSA and MRSP, we evaluated the occurrence and molecular characteristics of each in a veterinary college. MRSA and MRSP were isolated from nasal samples from veterinarians, staff members, and veterinary students affiliated with a veterinary hospital. Using stepwise logistic regression, we identified two factors associated with MRSA carriage: (i) contact with an identified animal MRSA case (odds ratio [OR], 6.9; 95% confidence interval [95% CI], 2.2 to 21.6) and (ii) being an employee (OR, 6.2; 95% CI, 2.0 to 19.4). The majority of MRSA isolates obtained from individuals affiliated with the veterinary hospital and dog patients harbored spa type t002 and a type II staphylococcal cassette chromosome mec (SCCmec), similar to the hospital-acquired MRSA isolates in Japan. MRSA isolates harboring spa type t008 and a type IV SCCmec were obtained from one veterinarian on three different sampling occasions and also from dog patients. MRSA carriers can also be a source of MRSA infection in animals. The majority of MRSP isolates (85.2%) carried hybrid SCCmec type II-III, and almost all the remaining MRSP isolates (11.1%) carried SCCmec type V. MRSA and MRSP were also isolated from environmental samples collected from the veterinary hospital (5.1% and 6.4%, respectively). The application of certain disinfection procedures is important for the prevention of nosocomial infection, and MRSA and MRSP infection control strategies should be adopted in veterinary medical practice.
A nationwide investigation of antimicrobial susceptibility in Escherichia coli isolated from food-producing animals was performed in Japan. MICs of 18 antimicrobial agents were determined for a total of 1018 isolates. Higher resistance rates were observed against sulfadimethoxine, oxytetracycline and dihydrostreptomycin, followed by ampicillin and kanamycin. Resistance was more frequently observed among broiler isolates, followed by isolates from pigs. Almost 10% of broiler isolates were resistant to fluoroquinolones and extremely high MICs (100 mg/L) were observed. In general, antimicrobial resistance rates in E. coli have declined in recent years, with the exception of resistance to fluoroquinolones among broiler isolates, which has increased.
Aims: To investigate the existing antimicrobial susceptibility and genetic characteristics of Lactococcus garvieae isolates from cultured Seriola in Japan.
Methods and Results: Minimum inhibitory concentrations (MICs) of 14 antimicrobial agents for 170 isolates were determined using the agar dilution method. Seventy‐five isolates (44·1%) were simultaneously resistant to erythromycin (EM) (MIC ≥ 2 μg ml−1), lincomycin (LCM) (MIC ≥ 128 μg ml−1) and oxytetracycline (OTC) (MIC ≥4 μg ml−1). Resistance to EM was grouped as intermediate‐ and high‐level resistant by MIC values. All resistant isolates possessed ermB and tet(S) genes. The number of different bands between pulsed‐field gel electrophoresis patterns of 25 isolates and two ATCC strains (isolated in 1974), determined using two enzymes (ApaI and SmaI), did not exceed 3.
Conclusions: The present resistance pattern observed with ermB and tet(S) is similar to that observed in previous reports. Moreover, the genetic characteristics of L. garvieae isolates from a wide area in Japan in 2002 and ATCC strains were closely related.
Significance and Impact of the Study: This study suggests that EM‐, LCM‐ and OTC‐resistant isolates have been present for 15 years and that L. garvieae strains with same origin have spread among Seriola spp. in Japan since 1974.
A total of 518 fecal samples collected from 183 apparently healthy cattle, 180 pigs and 155 broilers throughout Japan in 1999 were examined to determine the prevalence and antimicrobial susceptibility of Salmonella. The isolation rates were 36.1% in broilers, 2.8% in pigs and 0.5% in cattle. S. enterica Infantis was the most frequent isolate, found in 22.6% of broiler fecal samples. Higher resistance rates were observed against oxytetracycline (82.0%), dihydrostreptomycin (77.9%), kanamycin (41.0%) and trimethoprim (35.2%). Resistance rates to ampicillin, ceftiofur, bicozamycin, chloramphenicol and nalidixic acid were <10%. CTX-M-2 β-lactamase producing S. enterica Senftenberg was found in the isolates obtained from one broiler fecal sample. This is the first report of cephalosporin-resistant Salmonella directly isolated from food animal in Japan.
Aims: To compare the antimicrobial resistance, serotypes and flagellin gene types of Campylobacter isolated from humans and food‐producing animals and thereby facilitate elucidation of the origin of Campylobacter causing human infection in Japan.
Methods and Results: The MIC values of ampicillin, dihydrostreptomycin, gentamicin, erythromycin, oxytetracycline, nalidixic acid and enrofloxacin for Campylobacter isolated from humans (134 isolates), cattle (38 isolates), pigs (69 isolates), layers (84 isolates) and broilers (51 isolates) were compared. The MIC90 values of ampicillin for Campylobacter jejuni isolates from poultry were higher than those from humans and cattle. Campylobacter coli that was resistant to dihydrostreptomycin and erythromycin was observed at a higher frequency in humans and pigs than in poultry. The restriction fragment profiles of flaA of human, bovine and broiler isolates were analysed by clustering, and the isolates were classified into five clusters. Cluster I contained only human and bovine isolates. Clusters III, IV and V contained human, bovine and broiler isolates.
Conclusions: Campylobacter isolates from humans included isolates that exhibited characteristics identical to those of the bovine, porcine and poultry isolates.
Significance and Impact of the Study: In addition to poultry, cattle and pigs are believed to be sources of campylobacteriosis in Japan.
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