Association between dipeptidyl peptidase-4 inhibitors and autoimmune disorders: Data mining of the spontaneous reporting system in Japan.

The main objective of this study is to conduct a disproportionality analysis of adverse events in the Japan Adverse Event Report (JADER) database and evaluate the risk of the DPP-4 inhibitor induced autoimmune disorder, the secondary objective is risk assessment of sex difference and age difference. The proportional reporting ratio (PRR) of frequency-based statistics and Bayesian estimates of the information components (IC) were calculated as a measure of signal detection. Sex difference and age difference were evaluated using signal score calculated from the PRR and the Chi-square. In patients taking DPP-4 inhibitors, 94 reports of autoimmune disorders were detected with both signals; PRR: 4.09, chi-square: 158.26 and IC: 1.66, 95 % confidence interval: 1.32-2.00). For other antidiabetic drugs, no signals were detected. The signal of males was PRR: 4.53, chi-square: 110.91 and signal score: 6.22, the signal of female was PRR: 3.53, chi-square: 47.65 and signal score: 5.12. About age difference, the signal scores were 6.71 for patients over 60 years and 0.56 for patients under 60 years old. This study suggests that the DPP-4 inhibitors, unlike other antidiabetic drugs, were associated with autoimmune disorders. Signals of the DPP-4 inhibitors induced autoimmune disorders were detected in both male and female, but no sex difference was observed, but age difference was observed. Especially attention should be paid to patients over 60 years old.

Relationship Between Initial Renal Function and the Inhibitory Effect of Dipeptidyl Peptidase-4 Inhibitor Treatment on Renal Function Decline.

We retrospectively investigated the renal function index of patients with type 2 diabetes mellitus (T2DM) to examine the influence of dipeptidyl peptidase-4 (DPP-4) inhibitors on renal function between patients up to early nephropathy and after overt nephropathy. Patients with T2DM (>18 years old) who had been prescribed hypoglycemic agents for ≥3 months at Gifu Municipal Hospital between March 2010 and April 2014 were included in the study. Renal function was evaluated as the estimated glomerular filtration rate (eGFR) decline from baseline at 12 months. Patients in the DPP-4 inhibitor-treated and untreated groups with an eGFR ≥60 (358 [58.2 %] and 257 [41.8 %], respectively) and eGFR <60 (115 [60.2 %] and 76 [39.8 %], respectively) were subjected to multiple logistic regression analysis. Among patients with an eGFR ≥60, no significant differences were observed in eGFR decline rates over time. However, among patients with an eGFR <60, significant decreases were observed in eGFR decline rates >10 % (6 months; odds ratio, 0.476; P = 0.043, 12 months; odds ratio, 0.413; P = 0.010). Similar results were obtained for an eGFR decline rate >20 % (12 months; odds ratio, 0.369; P = 0.049). DPP-4 inhibitors are renoprotective in patients with T2DM and an eGFR <60.

Herd prevalence of Enterobacteriaceae producing CTX-M-type and CMY-2 ß-lactamases among Japanese dairy farms.

AIMS: To determine the herd prevalence of Enterobacteriaceae producing CTX-M-type extended-spectrum ß-lactamases (ESBLs) among 381 dairy farms in Japan. METHODS AND RESULTS: Between 2007 and 2009, we screened 897 faecal samples using BTB lactose agar plates containing cefotaxime (2 µg ml(-1)). Positive isolates were tested using ESBL confirmatory tests, PCR and sequencing for CTX-M, AmpC, TEM and SHV. The incidence of Enterobacteriaceae producing CTX-M-15 (n = 7), CTX-M-2 (n = 12), CTX-M-14 (n = 3), CMY-2 (n = 2) or CTX-M-15/2/14 and CMY-2 (n = 4) in bovine faeces was 28/897 (3·1%) faecal samples. These genes had spread to Escherichia coli (n = 23) and three genera of Enterobacteriaceae (n = 5). Herd prevalence was found to be 20/381 (5·2%) dairy farms. The 23 E. coli isolates showed clonal diversity, as assessed by multilocus sequence typing and pulsed-field gel electrophoresis. The pandemic E. coli strain ST131 producing CTX-M-15 or CTX-M-27 was not detected. CONCLUSIONS: Three clusters of CTX-M (CTX-M-15, CTX-M-2, CTX-M-14) had spread among Japanese dairy farms. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the prevalence of multidrug-resistant CTX-M-15-producing E. coli among Japanese dairy farms.

National surveillance of Salmonella Enteritidis in commercial eggs in Japan.

A total of 105 033 eggs were collected across Japan from June 2010 to January 2011 and tested for Salmonella Enteritidis to provide data for the risk profiling of S. Enteritidis in eggs by the Food Safety Commission of Japan. S. Enteritidis isolates were recovered from three samples (20 eggs/sample) and these samples were different in regard to sampling period, grading and packaging centre and farm. The prevalence of S. Enteritidis in commercial eggs in Japan is estimated at ~0.003% which was a tenfold decrease in prevalence compared to similar surveillance in the mid 1990s. The decrease in the contamination in commercial eggs is considered a contributory factor in the decrease of foodborne diseases associated with S. Enteritidis in this period.

Topical application of PPARα (but not ß/δ or γ) suppresses atopic dermatitis in NC/Nga mice.

BACKGROUND: Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors, which regulate not only adipogenesis and proliferation/differentiation but also the immune response of cells. Because topical application of the activators of some PPAR isoforms improved clinical symptoms in patients with atopic dermatitis (AD), we investigated the role of PPAR activators using a murine AD model in NC/Nga mice; to the best of our knowledge, this has not been previously reported. METHODS: Activators of three PPAR isoforms (α, ß/δ, γ) were topically applied on inflamed skin in a murine AD model that was developed by repeated topical application of mite antigen in NC/Nga mice. The efficacy of each topical PPAR activator was evaluated immunologically and serologically. RESULTS: Topical application of the PPARα activator, but not of the activators of PPARß/δ or PPARγ, improved clinical dermatitis, reduced inflammatory cell infiltration in the dermis, and alleviated the elevation of serum IgE levels. In addition, PPARα expression was downregulated in the epidermis in our murine AD model, as is seen in patients with AD. CONCLUSIONS: Topical application of PPARα activator could be a potent therapeutic agent for patients with AD and could take the place of topical steroid treatments.

T cell-specific overexpression of interleukin-27 receptor α subunit (WSX-1) prevents spontaneous skin inflammation in MRL/lpr mice.

BACKGROUND: Interleukin (IL)-27 and WSX-1, the receptor α-specific subunit, have been shown to play important roles in initiating Th1 responses and in inducing immune modulation, and the immunosuppressive effect of IL-27 appears to be exerted via suppression of IL-10 and IL-17, which may participate in the pathogenesis of human systemic lupus erythematosus (SLE). OBJECTIVES: To examine the significance of IL-27/WSX-1 signalling in spontaneous skin inflammation of MRL/lpr mice, a model for SLE. METHODS: The severity and development of skin lesions, dermal inflammatory cells and epidermal-dermal depositions in the skin lesions of MRL/lpr mice with CD2-promoted WSX-1 overexpression (WSX-1 Tg mice) and those with globally disrupted WSX-1 (WSX-1 KO mice) were examined and compared with those of MRL/lpr mice. RESULTS: By 4 months of age, both WSX-1 KO mice and control MRL/lpr mice developed predominantly similar skin inflammation, while WSX-1 Tg mice hardly did so, demonstrating that intensifying IL-27/WSX-1 signalling on T cells prevents the spontaneous skin inflammation. WSX-1 KO mice showed Th2-type skin inflammation as evidenced by the Th2-prone dermal infiltrating cells and an absence of cutaneous Th1-type IgG deposition. Interestingly, there were significant IL-17+ dermal infiltrating cells in both WSX-1 KO and control MRL/lpr mice, which might potentially contribute to the formation of skin inflammation in these mice. CONCLUSIONS: These data indicate that IL-27/WSX-1 signalling may play a protective role in the development of SLE-like skin inflammation, and modulating IL-27/WSX-1 signalling might be an interesting therapeutic strategy in the treatment of SLE.

Drug resistance and random amplified polymorphic DNA analysis of Photobacterium damselae ssp. piscicida isolates from cultured Seriola (yellowtail, amberjack and kingfish) in Japan.

AIMS: To investigate the antimicrobial susceptibility and genetic characteristics of Photobacterium damselae ssp. piscicida isolates obtained from cultured Seriola in Japan. METHODS AND RESULTS: Minimal inhibitory concentrations of 14 antimicrobials for 74 isolates from Seriola in Japan in 2002 were determined. Isolates showed high frequencies of resistance to sulfamonomethoxine (SMMX) (97.3%), oxytetracycline (OTC) (77.0%), flumequine (FMQ) (77.0%), chloramphenicol (CP) (75.7%), kanamycin (KM) (63.5%) and oxolinic acid (OA) (62.0%), but low to ampicillin (ABPC) (2.8%). All isolates were susceptible to bicozamycin (BCM), fosfomycin (FOM) and florfenicol (FF). Of these isolates, 45 (60.8%) showed same resistance pattern (SMMX-OTC-FMQ-OA-CP-KM). In random amplified polymorphic DNA (RAPD) analysis, no difference was observed among our 74 field isolates and ATCC51736 isolated from Seriola in 1974 in Japan, but different from ATCC 17911 isolated from white perch in USA. CONCLUSIONS: FF, BCM, FOM and ABPC were useful antimicrobials for treating pseudotuberculosis. However, the frequency of multidrug resistance was high. RAPD analysis showed homogeneity of isolates from Seriola in Japan. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that some antimicrobials were still useful for treating pseudotuberculosis and that P. damselae ssp. piscicida strains of same origin might have spread among Seriola in Japan since 1974.

Drug resistance and pulsed-field gel electrophoresis patterns of Lactococcus garvieae isolates from cultured Seriola (yellowtail, amberjack and kingfish) in Japan.

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>or=2 microg ml-1), lincomycin (LCM) (MIC>or=128 microg ml-1) and oxytetracycline (OTC) (MIC>or=4 microg 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.

Glutathione-S-transferase activity toward aflatoxin epoxide in livers of mastomys and other rodents.

In order to study the liver glutathione-S-transferase (GST) activity toward aflatoxin B1 (AFB1) epoxide in mastomys in comparison with other rodents, we performed in vitro studies of the cytosolic GST activity toward AFB1-epoxide using mastomys, rat, mouse and hamster liver. Also AFB1 metabolism by liver microsomes including formation of AFB1-DNA adducts was studied. Cytosolic GST activity toward AFB1-epoxide was highest in mastomys liver, and higher in the hamster and mouse livers than in the rat liver, correlating well with the differences of the sensitivity of these species to the toxicity of AFB1. However, no relationship was noted between the sensitivity of a given species to the toxicity of AFB1 and the microsomal activity of binding of AFB1 to DNA or metabolizing AFB1 to AFM1, AFQ1 and AFP1. These results demonstrate the importance of the GST mediated AFB1-epoxide conjugation with glutathione in determining the differing sensitivities of these species to AFB1 toxicity. The extremely high activity of GST in mastomys indicates that this species would be a good model animal for studying GST toward AFB1-epoxide.

New potent antioxidative o-dihydroxyisoflavones in fermented Japanese soybean products.

A potent antioxidative 6-hydroxydaidzein (6-OHD) was newly isolated from soybean koji fermented with Aspergillus oryzae. 6-OHD, in addition to 8-hydroxydaidzein and 8-hydroxygenistein, were found to be present in various fermented soybean products, including their koji. Considering that these o-dihydroxyisoflavones had strong antioxidative activities, they may contribute to protecting from oxidative deterioration during the processing of fermented soybean products.

[A study of the long-term effectiveness of an outpatient pulmonary rehabilitation program].

Pulmonary rehabilitation is one of the most important components of comprehensive care for patients with significant disability due to chronic respiratory failure. Because pulmonary rehabilitation has not been popular in Japan, the long-term effectiveness of pulmonary rehabilitation has rarely been reported. We therefore examined the long-term effectiveness of an outpatient pulmonary rehabilitation program for patients with chronic respiratory failure. Our program was composed of a once-a-week introduction program for 2 months and a support program that was continued every 4 weeks as long as possible. Thirty stable patients with chronic respiratory failure were enrolled in the program; 21 patients (COPD: 15, lung complications of tuberculosis: 6) completed the 9-week introduction program and the ensuing 6-month support program. Good compliance with the home training regimen was maintained during the period. The introduction program significantly alleviated dyspnea (Fletcher's grade: 3.3 to 3.0, p < 0.01) and improved the data for activity (Spector's score: 5.3 to 5.8, p < 0.01) and 6-minute walking distance (319 to 384 m, p < 0.01). These benefits were sustained during the 6-month support program. We concluded that outpatient pulmonary rehabilitation can alleviate dyspnea and improve the activity and exercise tolerance of patients with chronic respiratory failure, and that the effectiveness of training can be well maintained with a minimal support program.

Formation Mechanism for Potent Antioxidative o-Dihydroxyisoflavones in Soybeans Fermented with Aspergillus saitoi.

The formation mechanism for the potent antioxidative o-dihydroxyisoflavones, 8-hydroxydaidzein (8-OHD) and 8-hydroxygenistein (8-OHG), was studied by incubating whole soybeans in a solid culture and a soybean extract in a liquid culture with Aspergillus saitoi. Analyses of changes in the isoflavone analogue content, ß-glucosidase activity, and isoflavone hydroxylation ability indicated that 8-OHD and 8-OHG were formed from daidzein and genistein, respectively, by microbial hydroxylation, being respectively liberated from daidzin and genistin by ß-glucosidase from A. saitoi during incubation. No selective hydroxylation reaction at the 8-position of daidzein and genistein were apparent during the vegetative stage, but were induced at the stage of sporulation.

[A prospective study on the relationship between body mass index and mortality in middle-aged and elderly people in Japan].

We conducted a prospective study to clarify the relationship between mortality and body mass index (BMI) in a middle-aged and elderly population in Japan. The subjects included 12,649 people (5,686 males and 6,963 females) aged from 40-69 years, who were identified by a mail survey between 1987 and 1989, and thereafter were followed up from the date of the survey until February 28, 1995. Cox's proportional hazards model was used to adjust for age, smoking status, drinking habit and occupation, and hazard ratios were calculated for total mortality and cancer mortality based on levels of BMI, while using a BMI of 22 to approximately 24 as a reference level. Results showed a U shaped relationship between BMI and total mortality in females. Females demonstrated a minimal total mortality in the reference group, but a significantly high hazard ratio in the lowest group of BMI < 20 (hazard ratio = 1.95, p < 0.01) and the highest group of BMI > or= 26 (hazard ratio = 1.71, p < 0.01). On the other hand, males did not demonstrate such a U shaped relationship, but instead showed an L shaped relationship between BMI and total mortality. Males showed a minimal total mortality at a BMI of 24 of approximately 26 (hazard ratio = 0.92), but a significantly high hazard ratio in the lowest BMI group (hazard ratio = 1.57, p < 0.01). In contrast, an elevated hazard ratio was not observed in the highest BMI group (hazard ratio = 1.05). The relationship between BMI and cancer mortality was found to be similar to the relationship between BMI and total mortality in both males and females. Our findings suggest that moderate overweight may be a sign of good health among both middle-aged and elderly men in Japan.

Potent Antioxidative Isoflavones Isolated from Soybeans Fermented with Aspergillus saitoi.

Two potent antioxidative isoflavones were isolated from soybeans fermented with Aspergillus saitoi by silica gel column chromatography and preparative HPLC, using ODS column, or in addition, Toyopearl HW-40 column chromatography. The purified AS-13 and AS-9B compounds were identified as 8-hydroxydaidzein (8-OHD) and 8-hydroxygenistein (8-OHG), respectively, by MS, and (1)H-NMR, (13)C-NMR and HMBC spectra. These isoflavones, which have an o-dihydroxy structure between the 7- and 8-position, each exhibited significantly stronger antioxidative activity than daidzein and genistein in both oil and lipid/aqueous systems. Furthermore, the antioxidative activity and the content of each isoflavone analog in soybeans with different fermentation periods were investigated. It is suggested from these results that AS-13 and AS-9B were produced from daidzein and genistein, respectively, by hydroxylation at the 8-position of each isoflavone structure. In addition, it is concluded that these isoflavones were also the principal antioxidants in potent antioxidative soybeans fermented with A. saitoi.

Activity of H(+)-ATPase in ruminal bacteria with special reference to acid tolerance.

Batch culture experiments showed that permeabilized cells and membranes of Ruminococcus albus and Fibrobacter succinogenes, acid-intolerant celluloytic bacteria, have only one-fourth to one-fifth as much H(+)-ATPase as Megasphaera elsdenii and Streptococcus bovis, which are relatively acid tolerant. Even in the cells grown in continuous culture at pH 7.0, the acid-intolerant bacteria contained less than half as much H(+)-ATPase as the acid-tolerant bacteria. The amounts of H(+)-ATPase in the acid-tolerant bacteria were increased by more than twofold when the cells were grown at the lowest pH permitting growth, whereas little increase was observed in the case of the acid-intolerant bacteria. These results indicate that the acid-intolerant bacteria not only contain smaller amounts of H(+)-ATPase at neutral pH but also have a lower capacity to enhance the level of H(+)-ATPase in response to low pH than the acid-tolerant bacteria. In addition, the H(+)-ATPases of the acid-intolerant bacteria were more sensitive to low pH than those of the acid-tolerant bacteria, although the optimal pHs were similar.

Antioxidative mechanism and apoptosis induction by 3-hydroxyanthranilic acid, an antioxidant in Indonesian food Tempeh, in the human hepatoma-derived cell line, HuH-7.

Recently, a new potent antioxidant was isolated from Tempeh (a traditional fermented soybean food in Indonesia) and was identified as 3-hydroxyanthranilic acid (HAA). This study deals with the antioxidant mechanism of HAA under biological systems and the cytokilling function of HAA to human malignant cells. HAA eliminated free radicals and inhibited the formation of fatty acid hydroperoxide in vitro, suggesting that HAA would serve as an antioxidant in the initial reaction in lipid oxidation systems. Actually, HAA inhibited the formation of the dominant product of membrane lipids, 12-hydroxyeicosatetraenoic acid (12-HETE) at a high concentration, while HAA accelerated 12-HETE formation at a low concentration in mammalian tissue. HAA oxidized glutathione and inhibited superoxide dismutase in vitro. Furthermore, HAA inhibited cell growth and induced apoptosis to HuH-7, a human hepatoma-derived cell line. As long as HAA is taken as a component of Tempeh, and not in large doses as a chemical, it may possibly act as a prooxidant rather than an antioxidant in vivo.

Dengue hemorrhagic fever in Jakarta, Indonesia in 1988: isolation of dengue virus from patient whole blood using cell cultures.

During an outbreak of dengue hemorrhagic fever (DHF) in Jakarta, Indonesia in 1988, we attempted to isolate dengue virus using mosquito cells and a medium containing heparin. Whole blood, immediately after being drawn from patients, was inoculated into Aedes albopictus cell cultures temporarily maintained in the heparin-containing medium. The overall virus isolation rate was 25% (17 of 69) samples collected within three days after admission of the patients to hospital. No virus was obtained thereafter. The successful virus isolation was apparently not related to titers of anti-dengue virus hemagglutination-inhibiting antibodies present in patients' sera. The viruses were recovered from cases of each of the four World Health Organization grades of DHF without significant differences. The technique is simple and easily performed at bedside.

Characteristics of laryngeal receptors analyzed by presynaptic recording from the cat medulla oblongata.

In order to clarify the neural mechanisms for the protective laryngeal reflex, we conducted physiological analysis of laryngeal sensory receptors. In the present study, presynaptic unit activities, which might accurately reflect characteristics of the laryngeal receptor, were recorded with a glass microelectrode in the nucleus of the tractus solitarius of the medulla oblongata in ketamine-urethane anesthetized cats, and the responses to the mechanical and/or chemical stimuli were analyzed. From the results, it was demonstrated that highly sensitive mechanoreceptors and polymodal receptors exist in the laryngeal mucosa; they are particularly numerous in the laryngeal surface of the epiglottis and arytenoid region, and uncommon in the vocal fold. Mechanoreceptors on the laryngeal mucosa were classified into a rapidly adapting group and a slowly adapting group, while all polymodal receptors adapted rapidly to mechanical stimulation. These results suggest that these non-specific polymodal and rapidly adapting receptors may correspond to more superficial receptors such as free nerve endings and some taste buds, and also monomodal slowly adapting mechanoreceptors may correspond to deeper terminals in the subepithelium. It is also considered possible that the structures and the characteristics of these receptors are appropriate to elicit the protective laryngeal reflexes by non-specifically detecting various kinds of stimuli.

A prospective study on mortality among Japanese coal miners.

We conducted a prospective study to clarify mortality patterns among Japanese coal miners in a former coal mining area. Subjects included 1,796 coal miners and 4,022 non-coal-miners, who were identified by a mail survey between 1987 and 1989, and then followed up from the date of the survey to April 30th, 1994. We applied Cox's proportional hazards model to compare the mortalities between coal miners and non-coal-miners. Among the coal miners, significantly high risk ratios were observed in all causes of death (risk ratio = 1.4, p < 0.05) and all malignant neoplasms (risk ratio = 1.5, p < 0.05). Risk ratios for all causes of death and all malignant neoplasms also rose with the length of experience in coal mining. Analysis of the results for sites of cancer showed that coal miners had high risk ratios for stomach cancer (risk ratio = 1.6), liver cancer (risk ratio = 1.4) and lung cancer (risk ratio = 1.6), though these ratios were not statistically significant. When the risk ratio for lung cancer was analyzed according to the length of experience in coal mining, coal miners with at least 15 years' experience had a significantly high risk ratio (risk ratio = 2.4, p < 0.05), though coal miners with less than 15 years' experience had almost the same risk as non-coal-miners.