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Background Psoriasis is a chronic inflammatory skin disease. Anti-TNFα, IL-17A and IL-23p19 antibodies are effective for psoriasis. However, the contribution of regulatory T cells (Treg) in their effectiveness remains to be elucidated. Objective We investigated the effects of TNFα, IL-17A and IL-23p19 inhibition on Tregs in imiquimod-induced psoriasiform dermatitis. Methods Psoriasiform dermatitis was induced by imiquimod application on murine shaved back skin for six days. Mice were treated with anti-TNFα, IL-17A or IL-23p19 monoclonal antibodies every other day from one day before imiquimod application. Results Administration of anti-TNFα, IL-17A or IL-23p19 antibodies improved the clinical score and downregulated Th17-related cytokines and chemokines, while IL-23p19 antibodies upregulated IL-10 mRNA expression. Anti-IL-17A or IL-23p19 antibody-treated imiquimod-applied mice showed a significant increase in the number of Foxp3+ IL-10+ Tregs. Recipient mice adoptively transferred with Tregs derived from donor mice treated with antibodies demonstrated clinical and pathological improvement in imiquimod-induced psoriasiform dermatitis. Anti-IL-17A or IL-23p19 antibody-induced Tregs significantly increased the number of Foxp3+ cells and IL-10 expression in imiquimod-induced psoriasiform dermatitis in recipient mice but anti-TNFα antibody-induced Tregs did not. Conclusion Anti-IL-17A or IL-23p19 antibody inhibits the IL-17/IL-23 signaling pathway, and induces expansion of Tregs and their suppressive capacity in imiquimod-induced psoriasiform dermatitis.

Bullous pemphigoid (BP) is an acquired autoimmune blistering disease in which autoantibodies against epitopes in the basement membrane zone of the skin such as BP180 or BP230 are produced. Dipeptidyl peptidase (DPP)-4 inhibitors have become commonly used to treat diabetes. As DPP-4 inhibitors are more commonly prescribed for diabetes, BP related to DPP-4 inhibitors has been reported and has attracted attention. Therefore, we retrospectively investigated patients who were diagnosed with BP in order to examine characteristics of DPP-4 inhibitor-related BP (nine patients; median age, 85 years) in comparison with non-DPP-4 inhibitor-related BP (21; median age, 85 years). There was no significant difference in Bullous Pemphigoid Disease Area Index between DPP-4 inhibitor-related BP patients and non-DPP-4 inhibitor-related BP patients, except for erosions/blisters score in mucosa. Laboratory tests revealed no significant differences between DPP-4 inhibitor-related BP patients and non-DPP-4 inhibitor-related BP patients in total white blood cell count, eosinophil count, neutrophil count and the titer of anti-BP180 antibody. The number of eosinophils infiltrating into the skin was significantly lower in patients with DPP4 inhibitor-related BP than in patients with non-DPP4 inhibitor-related BP. Our results showed that DPP-4 inhibitor-related BP has some distinct pathological characteristics from BP not associated with DPP-4 inhibitor.

summary Conjunctivitis in patients with atopic dermatitis treated with dupilumab is associated with baseline serum levels of IgE and TARC but not subjective severity scores in a real-world setting. These objective parameters are useful as practical predictors of later development of conjunctivitis.

A screening test was undertaken to isolate microorganisms that produced ascorbate oxidase. The enzyme activity was found in a culture filtrate of a fungal strain (HI-25), newly isolated from a soil sample. Based on the morphological characteristics, this isolate was identified as Acremonium sp. From the examinations of cultural conditions, optimum conditions for enzyme production were found; strain HI-25 was aerobically cultured by a jar fermenter at 25°C in a medium containing 5% glycerol, 2% defatted soybeans, 0.1% monosodium L-glutamate, 0.1% KH2PO4, 0.02% MgSO4 ·7H2O, and 0.01% KCl, pH 6.0. After cultivation, an ascorbate oxidase was purified from the culture filtrate by an ammonium sulfate fractionation, column chromatographies on DEAE-cellulose and Butyl-Toyopearl, and gel filtration twice on Sephadex G-100. The purification was 850-fold with an activity yield of 8.8%. The purified enzyme gave a single band on SDS polyacrylamide gel electrophoresis, and had a molecular weight of 80,000 by SDS polyacrylamide gel electrophoresis and 76,000 by native gel filtration. This enzyme was most active at pH 4.0, 45°C, and was most stable between pH 6.0-10.0 and at temperatures below 60°C.

Staphylococcal exfoliative toxins are known to digest desmoglein (Dsg) 1, a desmosomal cell–cell adhesion molecule, thus causing intraepidermal splitting in human bullous impetigo, staphylococcal scalded skin syndrome and swine exudative epidermitis. Recently, a novel exfoliative toxin gene (exi), whose sequence shares significant homology with previously identified exfoliative toxins, was isolated from Staphylococcus pseudintermedius. Little is known about the pathogenic involvement of this toxin in canine pustular diseases such as impetigo. The aim of this study was to determine whether EXI, the product of the exi gene, digests canine Dsg1 and causes intraepidermal splitting in canine skin. An exi gene was isolated from chromosomal DNA of an S. pseudintermedius strain obtained from a pustule of a dog with impetigo, and was used to produce a recombinant EXI by Escherichia coli expression. When purified recombinant EXI was injected intradermally into normal dogs, it caused the development of vesicles or erosions with superficial epidermal splitting. In addition, the EXI abolished immunofluorescence for Dsg1, but not for Dsg3, at the injection sites. Moreover, the EXI directly degraded baculovirus-secreted recombinant extracellular domains of canine Dsg1, but not that of canine Dsg3, in vitro. The EXI also degraded mouse Dsg1α and swine Dsg1, but not human Dsg1, mouse Dsg1β and Dsg1γ. Conversely, recombinant SIET, previously designated as S. intermedius exfoliative toxin, did not cause intraepidermal splitting or degradation of any Dsgs. These findings indicate that EXI has a proteolytic activity that digests canine Dsg1, and this characteristic might be involved in the pathogenesis of intraepidermal splitting in canine impetigo. Resume Les toxines exfoliatives staphylococciques sont connues pour digerer la desmogleine (Dsg) 1, molecule d’adhesion intercellulaire, entrainant ainsi un clivage intra epidermique dans l’impetigo bulleux de l’homme, le “staphylococcal scalded-skin syndrome” et la dermatite exsudative porcine. Recemment, un nouveau gene de toxine exfoliative (exi), dont la sequence partage une homologie significative avec les toxines exfoliatives precedemment identifiees, a ete isolea partir de Staphylococcus pseudintermedius. Peu de choses sont connues sur le role pathogenique de cette toxine dans les maladies pustuleuses canines telle que l’impetigo. L’objectif de cette etude etait de determiner si EXI, le produit du gene exi, digere la Dsg1 canine et entraine un clivage intra-epidermique dans la peau canine. Un gene exi a ete isolea partir de l’ADN d’une souche de S. pseudintermedius preleve d’une pustule de chien atteint d’impetigo et a ete utilise pour produire un recombinant EXI par l’expression d’Escherichia coli. Lorsque le recombinant purifie a ete injecte en intradermique a des chiens normaux, il a cause le developpement de vesicules ou d’erosions avec clivage epidermique superficiel. De plus, EXI a aboli l’immunofluorescence pour Dsg1, mais pas pour Dsg3, aux sites d’injections. En outre, EXI a degrade directement les domaines extracellulaires secretes par les baculovirus de Dsg1 canine, mais pas ceux de Dsg3 canine in vitro. EXI a egalement degrade Dsg1α de souris et Dsg1 porcine, mais pas Dsg1 humaine, Dsg1 β et Dsg1γ murines. Inversement, SIET recombinant, precedemment presentee comme une toxine exfoliative de S. intermedius, ne causait pas de clivage intra epidermique ou de degradation d’aucune des Dsg. Ces donnees indiquent que EXI a une activite proteolytique qui digere la Dsg1 canine et cette caracteristique pourrait etre impliquee dans la pathogenie du clivage intra-epidermique de l’impetigo canin. Resumen Las toxinas estafilocicas exfoliativas digieren la desmogleina-1 (Dsg-1), una molecula de adhesion intercelular presente en desmosomas, lo cual causa separacion intraepidermal en el impetigo bulloso humano, el sindrome estafilococico de piel escaldada y la epidermitis exudativa porcina. Recientemente se ha aislado un nuevo gen de una toxina exfoliativa (exi) en Staphylococcus pseudintermedius. Este gen es bastante homologo en secuencia con toxinas exfoliativas previamente caracterizadas. Se sabe poco acerca de la influencia de este gen en la patogenia de las enfermedades pustulares de la piel en perros, como el impetigo. El proposito de este estudio fue determinar si EXI, el producto del gen exi, digiere la Dsg1 canina y causa separacion intraepidermal en la piel de perros. Se aislo un gen exi de un DNA cromosomico de una cepa de S. pseudointermedius obtenida de una pustula de un perro con impetigo, y se utilizo para producir EXI recombinante expresado en E. coli. Cuando se inyecto la proteina EXI purificada por via intradermal a perros sanos, causo el desarrollo de vesiculas o erosiones con separacion de la epidermis superficial. Ademas la proteina EXI inhibia la inmunofluorescencia frente a Dsg1, pero no para Dsg3, en los lugares de inyeccion. Lo que es mas, in vitro EXI directamente degradaba dominios extracelulares de Dsg1 producidos en baculovirus, pero no los de Dsg3. La proteina EXI tambien degradaba Dsg1α de raton y Dsg1 de cerdo, pero no la Dsg1 de humanos, ni las Dsg1β y Dsg1γ de raton. Por el contrario, SIET recombinante, previamente presentada como una toxina exfoliativa de S. intermedius, no producia separacion intraepidermal o degradacion de ninguna de la Dsgs. Estos hallazgos indican que EXI tiene actividad proteo litica que digiere la Dsg1 y esta propiedad puede estar implicada en la patogenesis de la separacion intraepidermal en el impetigo canino. Zusammenfassung Exfoliative Staphylokokkentoxine verdauen Desmoglein (Dsg) 1, ein desmosomales Zell-Zell Adhasionsmolekul, und verursachen dadurch ein intraepidermales Splitting beim humanen bullosen Impetigo, beim “Staphylokokken Scalded Skin Syndrom” und bei der exudativen Epidermitis des Schweines. Unlangst wurde das Gen eines neuen exfoliativen Toxins (exi), dessen Sequenz eine signifikante Homologie mit schon fruher identifizierten exfoliativen Toxinen zeigt, von Staphylokokkus pseudointermedius isoliert. Es ist wenig bekannt, welche pathologische Rolle dieses Toxin bei pustulosen Erkrankungen des Hundes, wie Impetigo, spielt. Das Ziel dieser Studie war es zu bestimmen, ob EXI, ein Produkt des exi Gens, canines Dsg1 verdaut und intraepidermales Splitting in der Hundehaut verursacht. Ein exi Gen wurde aus chromosomaler DNA eines S. pseudointermedius Stammes, der von einer Pustel eines Hundes mit Impetigo stammte, isoliert. Dieses wurde verwendet, um ein rekombinantes EXI mittels Expression in Escherichia coli zu produzieren. Wenn aufbereitete rekombinante EXI intradermal in normale Hunde injiziert wurden, verursachte dies die Entstehung von Blaschen oder Erosionen mit oberflachlichem epidermalen Splitting. Zusatzlich hob EXI an den Injektionsstellen die Immunfluoreszenz fur Dsg1, jedoch nicht fur Dsg3 auf. Daruber hinaus schwachte EXI in vitro direkt die durch ein Bakulovirus-System exprimierten rekombinanten extrazellularen Domanen von caninem Dsg1, aber nicht von caninem Dsg3. EXI schwachte auch murine Dsg1α und porcine Dsg1, aber nicht humanes Dsg1, murines Dsg1β und Dsg1γ. Im Gegensatz dazu verursachte rekombinantes SIET, welches schon fruher als S. intermedius exfoliatives Toxin eingefuhrt wurde, weder intraepidermales Splitting noch eine Abschwachung von Dsgs. Diese Ergebnisse weisen auf die proteolytische Aktivitat von EXI hin, welches canines Dsg1 verdaut und diese Eigenschaft konnte an der Pathogenese des intraepidermalen Splittings bei caninem Impetigo beteiligt sein.

Activation of anti-apoptotic gene transcription by NF-κB (nuclear factor-kappa B) has been reported to be linked with a resistance of cancer cells against chemotherapy. NEMO (NF-κB essential modulator) interacts with a number of proteins and modulates the activity of NF-κB pathway. In this study, we revealed that RPAP3 (RNA polymerase II-associated protein 3) possesses an activity to bind with NEMO and to inhibit the ubiquitination of NEMO and that RPAP3 enhances doxorubicin-induced cell death in breast cancer cell line T-47D through the marked impairment of NF-κB pathway. These results indicate that RPAP3 may be a novel modulator of NF-κB pathway in apoptosis induced by anti-cancer chemotherapeutic agents.

The resorption of alveolar bone in periodontitis and/or following tooth extraction results in a significant problem in an individual's oral health-related quality of life. Strategies employing growth factor proteins and/or stem cell-based therapy have been successfully used to regenerate alveolar bone augmentation/regeneration; however, cost-effectiveness issues illustrate the need for developing alternative strategies. One promising approach is the development of conventional chemical and biological therapeutics to strongly stimulate endogenous cells to regenerate. Small-molecule compounds can have a variety of biological functions, serving as cell signaling molecules, as tools in molecular biology, and as drugs in medicine. Therefore, the discovery of natural and novel small-molecule synthetic regulators of differentiation and maturation of osteoblasts and/or osteoclasts would accelerate the development of bone regenerative medicine. Recently, "chemical biology" has started to play an increasingly important role in both drug discovery and elucidating the mechanism of biological phenomena. This review first addresses the current needs and strategies for alveolar bone regeneration, followed by the potential small molecules associated with bone regeneration, with a concluding section on chemical biology-based high throughput screening approaches for identifying small molecules targeting osteoblast/osteoclast differentiation.

We characterized 53 Salmonella enterica ser. Typhimurium strains recovered from healthy pigs during 1998-1999 (n=12) and 2004-2005 (n=41) as to their carriage of DT104 spacer region, class 1 and 2 integrons, virulence genes (spvC, rck, and pefA), and XbaI- and BlnI-Pulsed-Field Gel Electrophoresis (PFGE) profiles. No DT104 strain was detected in 1998-1999, whereas 65.9% (27/41) of the strains in 2004-2005 were DT104 showing resistance to ampicillin, chloramphenicol, streptomycin, sulfonamides, tetracycline, and cephaloridine (R-type ACSSuT+). Class 1 intergron-associated genes, aadA2 (1.0-kbp amplicon) and pse1 (1.2-kbp amplicon), were found in all the DT104 strains (27/27). One strain showing resistance to streptomycin, sulfonamides, tetracyclin, and trimethoprim (R-type SsuTTm) harbored another class 1 integron-associated gene (dhfrXII-orfF- aadA2) on 1.9-kbp amplicon. Virulence gene spvC was found in 92.5% (49/53) and rck and pefA were found in 88.7% (47/53) of the strains, whereas spvC, rck, and pefA were found in all the DT104 strains. Ser. Typhimurium strains were categorized into four clusters (X1, X2, X3, and X4a/X4b) by XbaI-PFGE, or into nine clusters (B1, B2, B3a/B3b, B4, B5, B6, B7, B8, B9a/B9b) by BlnI-PFGE analyses. DT104 strains were restricted into X2, or into B2, B3a/B3b, and B6 clusters, indicating that our multidrug-resistant DT104 strains from healthy pigs might have derived from at least three independent clones, with the most widespread clone being the cluster B6 strains isolated in Kanto, Tokai, Chugoku, and Kyushu regions.

Staphylococcus aureus, Staphylococcus hyicus, and Staphylococcus chromogenes are known to cause skin infections in human or animals by producing exfoliative toxins (ETs). Staphylococcus pseudintermedius can also cause canine pyoderma, but no exfoliative toxins or similar toxins have been reported. PCR with degenerate primers targeted to the conserved regions in ETA, ETB, and ETD from S. aureus and SHETB from S. hyicus, and subsequent chromosome walking identified a novel gene, designated as exi (exfoliative toxin of pseudintermedius) in S. pseudintermedius. EXI had significant homologies with the exfoliative toxins (43-68% identity), particularly with ETB (67.1%), ETD (67.9%), and SHETB (65.1%). Phylogenetic analysis showed close relation between EXI and ETB with a bootstrap value of 80%. Neonatal mice injected with the crude proteins from the culture supernatant or recombinant EXI showed gross blisters and/or characteristic skin exfoliation. The prevalence of exi assessed by dot-blot hybridization was 23.3% (10/43) in S. pseudintermedius isolates from canine pyoderma. The EXI reported herein is the first exfoliative toxin identified in S. pseudintermedius.

Staphylococcus intermedius isolates (n=106), including 44 dog isolates and 62 pigeon isolates, were examined for their susceptibility to ampicillin, cephalexin, erythromycin, gentamicin, kanamycin, lincomycin, norfloxacin, oxacillin, tetracycline, and vancomycin by standard disk-diffusion test. The frequencies of resistance to ampicillin, kanamycin, and tetracycline were significantly higher in dog isolates than pigeon isolates (95.5% vs. 0%, 31.8% vs. 0%, and 45.5% vs. 9.7%, respectively; P

SUMMARYTo determine prevalence, serotype diversity and antimicrobial resistance of Salmonella in healthy pigs, faecal samples from 6771 pigs on 73 farms collected during 1998–1999 and 2004–2005 were examined. Salmonella isolates were serotyped and tested for susceptibility to 22 antimicrobials: benzylpenicillin, ampicillin, amoxicillin, cefazolin, cephaloridine, gentamicin, kanamycin, streptomycin, fradiomycin, colistin, tetracycline, chlortetracycline, oxytetracycline, chloramphenicol, thiamphenicol, sulfadimethoxine, sulfamethoxazole, sulfamethoxypyridazine, trimethoprim, sulfamethoxazole/trimethoprim, norfloxacin and ofloxacin. Farm-level and pig-level Salmonella prevalences were 35·5% and 2·2% in 1998–1999, and 35·7% and 3·3% in 2004–2005. Prevalence by growth stage was 2·4% for sows, 3·3% for weaned pigs, 2·7% for fattening pigs and 3·8% for finishing pigs. The predominant serotypes identified were Agona (28·4%), Typhimurium (17·9%) and Infantis (16·4%) in 1998–1999, and Typhimurium (32·5%), Anatum (24·6%) and Infantis (13·5%) in 2004–2005. Compared with the 1998–1999 isolates, the 2004–2005 isolates showed significantly higher rates of resistance to all the antimicrobials except tetracyclines (PPvs. 39·7% (50/126), PSalmonella prevalence in healthy pigs of different growth stages in Japan.

In view of the recent rapid increase in incidence of infection with antimicrobial resistant bacteria in human medicine, there is international controversy as to the medical risk that is created by transfer of antimicrobial resistant bacteria and antimicrobial resistant genes, which may be produced through the processes of administration of antimicrobials to food-producing animals, via the food chain. Accordingly, International Cooperation on Harmonization of Technical Requirements for Registration of Veterinary Medicinal Products (VICH) provides the comprehensive guidelines for characterizing selection of antimicrobial resistance by bacteria which may adversely affect human health in order to establish the system of registry of antimicrobial drugs to be administered to food- producing animals. Currently, Japanese Society of Antimicrobials for Animals recognizes the lack of technical test standards in compliance with VICH guidelines in the world and has established the test standards for "in vitro mutation frequency studies" to evaluate the appearance frequency and resistant level of resistant bacteria and those for studies on the "antimicrobial activity in gut" to estimate the effects on intestinal flora from professional aspects.

Staphylococcus intermedius isolates from dogs ( n = 44) and pigeons ( n = 62) were categorized into 12 types by intergenic ribosomal DNA spacer polymorphism analysis. All isolates from pigeons were lukS positive and all isolates from dogs were lukS and lukF positive by dot blot analysis. The mean leukotoxicity titer for dog isolates was at least 129-fold higher than that for pigeon isolates.

Mycobacterium marinum is difficult to distinguish from other species of Mycobacterium isolated from fish using biochemical methods. Here, we used genetic and proteomic analyses to distinguish three Mycobacterium strains: M. marinum strains MB2 and Europe were isolated from tropical and marine fish in Thailand and Europe, and Mycobacterium sp. 012931 strain was isolated from yellowtail in Japan. In phylogenetic trees based on gyrB, rpoB, and Ag85B genes, Mycobacterium sp. 012931 clustered with M. marinum strains MB2 and Europe, but in trees based on 16S rRNA, hsp65, and Ag85A genes Mycobacterium sp. 012931 did not cluster with the other strains. In proteomic analyses using a Bruker matrix-assisted laser desorption ionization Biotyper, the mass profile of Mycobacterium sp. 012931 differed from the mass profiles of the other two fish M. marinum strains. Therefore, Mycobacterium sp. 012931 is similar to M. marinum but is not the same, suggesting that it could be a subspecies of M. marinum.

Small molecule compounds that potently affect osteoclastogenesis could be useful as chemical probes for elucidating the mechanisms of various biological phenomena and as effective therapeutic strategies against bone resorption. An osteoclast progenitor cell-based high-throughput screening system was designed to target activation of NFAT, which is a key event for osteoclastogenesis. Orphan ligand library screening using this system identified the β-carboline derivative harmine, which is a highly potent inhibitor of dual-specificity tyrosine-phosphorylation regulated kinase 1A (DYRK1A), to be an NFAT regulator in osteoclasts. RAW264.7 cells highly expressed DYRK1A protein, and in vitro phosphorylation assay demonstrated that harmine directly inhibited the DYRK1A-mediated phosphorylation (in-activation) of NFATc1. Harmine promoted the dephosphorylation (activation) of NFATc1 in RAW264.7 cells within 24h, and it significantly increased the expression of NFATc1 in RAW264.7 cells and mouse primary bone marrow macrophages (BMMs) both in the presence and absence of RANKL stimulation. Although harmine promoted NFATc1 expression and stimulated target genes for osteoclastogenesis, cell-cell fusion and the formation of TRAP-positive multinucleated osteoclasts from RAW264.7 cells and BMMs was significantly inhibited by harmine treatment. Meanwhile, harmine remarkably promoted the expression of inhibitor of DNA binding/differentiation-2 (Id2), which is a negative regulator for osteoclastogenesis, in RAW264.7 cells and BMMs. An Id2-null-mutant showed slightly increased osteoclast formation from BMMs, and the harmine-mediated inhibition of osteoclast formation was abolished in the BMMs of Id2-null-mutant mice. These results suggest that harmine is a potent activator of NFATc1 that interferes with the function of DYRK1A in osteoclast precursors and also up-regulates Id2 protein, which may dominantly inhibit expression pathways associated with cell-cell fusion, thereby leading to the disruption of the fusion events mediating osteoclastogenesis. The small molecule harmine is therefore expected to provide an experimental tool for investigating signaling cascades in osteoclastogenesis, especially those centered on DYRK1A-mediated NFATc1 and Id2 regulation.

l-Iduronic acid-type 1-N-iminosugars, (3R,4S,5R,6R)- and (3R,4S,5S,6R)-6-acetamido-4-amino-5-hydroxypiperidine-3-carboxylic acid (6 and 7, respectively), (3R,4S,5R,6R)-6-acetamido-4-guanidino-5-hydroxypiperidine-3-carboxylic acid (8), and (3R,4S,5R,6R)-4-amino- and -guanidino-5-hydroxy-6-(trifluoroacetamido)piperidine-3-carboxylic acid (9 and 10, respectively), were synthesized from siastatin B (1), isolated from Streptomyces culture, by the intramolecular Michael addition of O-imidate to its α,β-unsaturated ester through cis oxiamination as a key step. Preincubation of B16 BL6 cells with these compounds inhibited invasion of the cells through reconstituted basement membranes. Pulmonary metastasis of B16 BL6 cells in mice was remarkably inhibited by pretreatment of the cells with these compounds in culture.

Staphylococcus hyicus isolates (n=207), including 150 exfolitative toxigenic and 57 non-toxigenic strains, were examined for their susceptibility to 13 antimicrobial agents by using the dehydrated 96-well MIC panel system. The frequency of their resistance to penicillin and ampicillin was 76.8% (159/207), followed by erythromycin (56%, 116/207), trimethoprim-sulfamethoxazole (28.5%, 59/207), chloramphenicol (24.2%, 50/207), kanamycin (19.8%, 41/207), and doxycycline (1.4%, 3/207). Resistance to chloramphenicol and trimethoprim-sulfamethoxazole was significantly higher in toxigenic strains than non-toxigenic strains (p

We investigated the carriage of Staphylococcus hyicus and their exfoliative toxin genes (exhA, exhB, exhC, exhD, and shetb) among 424 pigs from 38 Japanese pig farms by PCR and characterized the isolates by 16S-23S intergenic spacer region polymorphism analysis. S. hyicus (n=207) were isolated from 17.9% of pigs and 72.5% of them were toxigenic. The isolation rate of toxigenic S. hyicus was four times higher in the pigs with exudative epidermitis than the healthy pigs (87.6% versus 19.6%; p

Background Staphylococcus intermedius has been isolated from healthy dogs and pigeons as well as diseased dogs. Similar to Staphylococcus aureus, S. intermedius is known to carry many virulence factors but most of these factors remain to be studied. In this study, we examined 106 S. intermedius isolates (44 dog isolates and 62 pigeon isolates) for their hemolytic activity, biofilm formation, protease activity, and clumping factor and protein A production. Results Forty-three dog isolates (97.7%) and all pigeon isolates were hemolytic on sheep RBCs with a mean hemolytic titer of 336.7 and 47.32, respectively, whereas 43 dog isolates (97.7%) and 11 pigeon isolates (17.7%) exhibited a significant difference in their hemolytic activity on rabbit RBCs with a mean hemolytic titer of 11.04 and 3.76, respectively (p < 0.0005). The mean biofilm formation activity for dog isolates was 0.49, which was significantly higher than that (0.33) for pigeon isolates (p < 0.0005). Twenty-four dog isolates (54.5%) and 11 pigeon isolates (17.7%) were protease positive. Twenty-four dog isolates (54.5%) were clumping factor- and protein A- positive. Conclusion S. intermedius strains carrying the virulence factors examined in this study were more prevalent in dogs than pigeons.

Aims: To determine the prevalence of enterotoxin-producing Staphylococcus intermedius in dogs and pigeons. Methods and Results: A total of 106 S. intermedius isolates from 44 dogs and 62 pigeons were tested for the production of enterotoxins A, B, C and D by reverse passive latex agglutination (RPLA) and for sec-canine by PCR. Only one isolate from dog was positive for SEC and sec-canine. Screening of sec-canine-negative strains by nested PCR led to the identification of a novel enterotoxin-related gene, se-int. SE-int showed a significant homology (59–61% identity) with SEC and (56·6% identity) SEB. All 44 isolates from dogs and five isolates (8·1%) from pigeons were se-int positive. Conclusions: While S. intermedius was isolated more frequently from pigeons than from dogs, se-int was more prevalent among the S. intermedius isolates from dogs, compared with the pigeon isolates. Significance and Impact of the Study: Further characterization of the se-int-positive S. intermedius strains should clarify their pathogenic potential including enterotoxigenicity and zoonotic transmissibility to human beings.

We examined the dynamics of antibiotic resistance and pulsed-field gel electrophoresis (PFGE) patterns of E. coli isolates from health swine feces. Ten piglets were randomly chosen in each of 4 pig-raising farms. Feces were collected at the piglet stage, growing stage and at the finishing stage. Ten strains were isolated from each sample and isolates identified as E. coli were used for analysis. All the 553 strains, isolated at the piglet and growing stage of 3 farms, showed resistance against 2 to 9 antibiotics. Strains isolated at the finishing stage showed reduction of antibiotic resistance, and 70 of 294 strains showed susceptibility for all the 15 antibiotics tested. Similar reduction was observed in the remaining farm. However, in this farm, antibiotic susceptible strains were isolated from all stages, being 62 of 300 strains susceptible for all the antibiotics. The majority of strains showing the same antibiotic resistance pattern showed the same PFGE pattern, but also different PFGE patterns were found among those strains, even they have been isolated from the same animal.These results demonstrated that, the E. coli strains of each farm might be originated from multiple sources, and, although a same strain could remain along the stages, changes of antibiotic resistance patterns and PFGE profiles occurred in a short time period. Thus, for monitoring systems related to food safety, analysis at the finishing stage seems the most recommended. Moreover, studies to elucidate the causes of changes in the E. coli strains of intestinal flora might be of great interest.

L-Iduronic acid-type 1-N-iminosugars, (3R,4S,5R,6R)- and (3R,4S,5S,6R)-6-acetamido-4-amino-5-hydroxypiperidine-3-carboxylic acid (6 and 7, respectively), (3R,4S,5R,6R)-6-acetamido-4- guanidino-5-hydroxypiperidine-3-carboxylic acid (8), and (3R,4S,5R,6R)-4-amino- and -guanidino-5-hydroxy-6-(trifluoroacetamido) piperidine-3-carboxylic acid (9 and 10, respectively), were synthesized from siastatin B (1), isolated from Streptomyces culture, by the intramolecular Michael addition of O-imidate to its alpha,beta-unsaturated ester through cis oxiamination as a key step. Preincubation of B16 BL6 cells with these compounds inhibited invasion of the cells through reconstituted basement membranes. Pulmonary metastasis of B16 BL6 cells in mice was remarkably inhibited by pretreatment of the cells with these compounds in culture.

(1991). An Acidic Ascorbate Oxidase Produced by Acremonium sp. HI-25. Agricultural and Biological Chemistry: Vol. 55, No. 6, pp. 1693-1694.

屠場残渣(以下SR)を飼料資源として利用する目的で好気発酵処理を施し,豚を用いて発酵屠場残渣の嗜好性を調べた.好気発酵処は曝気量を変え,発酵温度,水分,pH,有機物量,微生物などの変動について調べた.そして,発酵屠場残渣(以下FSR)の飼料成分について分析し,さらに豚での嗜好性について検討した.以下にその結果を示す.1. SRを発酵する場合,適正曝気量はSR100kgあたり10l/minであった.2. SRの発酵熱によって大腸菌群やサルモネラ属菌は減少した.3. FSRは粗蛋白質や粗脂肪を比較的多く含んでいた.4. 市販配合飼料にFSRを60%まで混合しても残飼は認められず,豚での嗜好性は良好であった.

Rat brain slices were incubated with substance P (SP), and the SP receptors on the membranes from those slices were characterized by a 3H-SP binding technique. The number of substance P receptors measured in the extensively washed membrane preparations pretreated with 3 × 10−5M SP was reduced by 30% compared with that in nontreated membranes. This reduction was dependent on the incubation time and temperature. The metabolic inhibitors sodium azide and 2,4-dinitrophenol protected SP receptors from the reduction. The characteristics of 3H-SP incorporation into rat brain slices were similar to those of SP receptor down-regulation, that is, the 3H-SP incorporation was time, temperature, and energy dependent. Thus these results indicate that the processes of ligand incorporation and receptor down-regulation are closely associated phenomena. These observations may be important in elucidating the phenomenon of SP-induced desensitization.

Transmissibility of dihydrostreptomycin (DHSM) resistance in Pseudomonas lachrymans was studied. Transmission experiments were conducted by mixing-culture method, and it was found that the resistance of 13 P. lachrymans isolates out of 109 DHSM-resistant ones was distinctly transmitted to the recipient isolate of rifampicin-resistant P. lachrymans. Transmission frequency remarkably increased with the lapse of time of mixing-culture, and it was 10-1 at after 24hr. DHSM resistance was not transmitted when culture filtrate of one of the 13 DHSM-resistant isolates was used as donor, suggesting the presence of resistance (R) plasmid in the donor isolate. Several curing agents such as acriflavine, acridine orange etc. were tested against the donor resistant isolate, but the results were negative. From the resistant isolate and R+ transconjugant obtained from it, plasmid DNA was extracted with cleared lysate method and analyzed by agarose gel electrophoresis. From these experiments, it was clarified that the resistant isolate carries at least 2 kinds of plasmid DNA, molecular weights of which were 86×106 and 58×106 daltons. These plasmid DNAs could not be found in the transconjugant. Moreover, transformation experiments by these plasmid DNA samples on DHSM-susceptible P. lachrymans, P. aeruginosa and Escherichia coli were not successful. It seems, therefore, that these 2 kinds of plasmid DNA are unrelated to DHSM resistance, but interpreted as the so called cryptic plasmids.

The in vitro antifungal activity of ME1401, a potential topical antifungal agent, was compared with that of haloprogin, clotrimazole, miconazole, tolnaftate, and ciclopirox olamine by using an agar dilution procedure. ME1401 showed a broad antifungal spectrum and inhibited all of the 428 strains of 52 species of pathogenic yeasts and filamentous fungi tested at concentrations ranging from 0.01 to 12.5 micrograms/ml. In general, the activity of ME1401 was comparable or superior to that of clotrimazole and was greater than that of haloprogin and the other reference drugs under the conditions used. Only tolnaftate was superior to ME1401 in its activity against clinical isolates of Trichophyton rubrum. ME1401 showed no cross-resistance with any of the reference drugs and exhibited potent fungicidal activity.

Modes of actions of antitumor agents can be predicted by comparison of antitumor activities against L1210 cells in the presence and absence of some nucleosides. 1) Deoxyribonucleic acid (DNA) inhibitors were mainly antagonized by thymidine. 2) Both DNA and ribonucleic acid (RNA) inhibitors were mainly antagonized by adenosine and uridine. 3) Uridine antagonized the activities of an RNA inhibitor. 4) None of these nucleosides tested antagonized the antitumor activities of an inhibitor of protein synthesis. 5) A mitotic inhibitor was weakly antagonized by thymidine, adenosine and uridine. This method is useful for approximating the modes of actions of antitumor agents without using radioactive precursors.

Bordetella bronchiseptica strains isolated from the nasal cavities of young pigs in Japan from 1969 to 1972 were surveyed for drug resistance and distribution of R factors. Of 304 strains examined, 71 (23%) were resistant to either one or more of following three drugs, streptomycin (SM), sulfadimethoxine (SA), and aminobenzyl penicillin (APC). Triple (SM.SA.APC)-resistance was most frequent among these resistant strains. Strains of double (SM. SA)- or single (SM)- and (SA)-resistance were also isolated, but were very few in numbers. Of the 71 drug-resistant strains, 61 (86%) were found to carry R factors which were capable of conjugal transfer. All of these R factors had the triple (SM.SA.APC)-resistant markers and were identified as fi– (no fertility inhibition) type. The (SM.SA.APC)-resistant strains carrying R factors had been isolated from pigs reared on various farms in different districts, and consequently the prevalence of B. bronchiseptica strains carrying R factors was considered to be relatively wide-spread in young pigs.

Hylambatin (Hyl), a dodecapeptide isolated from the skin of the African frog, Hylambates maculatus, belongs to the family of tachykinin or physalaemin-like peptides. Hylambatin and its 12 fragments were tested in the guinea-pig ileum preparation for contractile activities. All fragments except 3 had contractile activities. The C-terminal fragment as short as the octapeptide sequence was at least as active as the parent molecules. The heptapeptide fragment (Hyl6-12) and the hexapeptide fragment (Hyl7-12) were less active and the C-terminal pentapeptide fragment (Hyl8-12) and the N-terminal hexapeptide fragment (Hyl1-6) were much less active. The N-terminal pentapeptide fragment (Hyl1-5) and the N-terminal fragment from which the N-terminal Asp or Asp-Pro residues were removed (Hyl2-6, Hyl3-6), were inactive at doses used.