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In this study, the distribution of hygienic indicator bacteria in cattle livers and bile was examined at slaughterhouses. One hundred twenty-seven cattle livers with gallbladders were carefully eviscerated from carcasses at 10 slaughterhouses. Microbiological examination revealed that nine bile samples (7.1% prevalence) and 19 liver parenchyma samples (15.0% prevalence) were positive for Enterobacteriaceae (EB) with means ± standard deviations of 3.68 ± 4.63 log CFU/mL and 1.59 ± 2.47 log CFU/g, respectively; thus, bacterial contamination was apparent even at the postevisceration stage. Subsequently, 70 cattle livers were obtained at the postprocessing and storage stage from 7 of the 10 slaughterhouses. Microbiological analysis revealed significantly higher levels of EB in the liver parenchyma (3.00 ± 3.89 log CFU/g, P = 0.011) than those at the postevisceration stage, suggesting that bacterial dissemination and/or replication occurred in the liver parenchyma during processing and storage. According to 16S rRNA ion semiconductor sequencing analysis of representative samples from 12 cattle, Proteobacteria, Firmicutes, and Actinobacteria were dominant in both the parenchyma and bile in which EB and Escherichia coli were predominant among livers with higher EB levels. These results suggest that bile plays a role as a vehicle for bacterial transmission to the liver parenchyma. This study is the first to evaluate bacterial distribution and community structure in the liver and biliary microecosystem of cattle at slaughter. Our data support the use of EB testing of bile to screen cattle livers contaminated with high levels of fecal indicator bacteria.

Cronobacter spp. cause serious diseases, such as necrotizing enterocolitis, bacteremia, and meningitis in neonates and infants. Most Cronobacter-associated meningitis is reportedly due to C. sakazakii and the majority of infections caused by C. malonaticus occur in adults and are less severe. We report the case of meningitis and brain abscess caused by C. malonaticus Sequence Type (ST) 440 in a healthy full-term neonate. We should consider the possibility that full-term neonates may develop meningitis due to C. malonaticus and treat appropriately because its mortality rate is very high, and survivors are usually left with severe neurologic impairment. In addition, C. malonaticus ST440 may have virulence factors that cause neonatal meningitis akin to the previous report of meningitic ST307 strain.

Unilaterally swollen eyes were histopathologically characterized in four MG-W gerbils. The primary lesions resided in the anterior segment of the eye where neural crest cells play a critical role in embryonic development. They included indistinct filtration angle, unformed canal of Schlemm, hypoplastic iris, and ciliary body. The findings noted in the retina, optic nerve, optic tract, and lateral geniculate nucleus were consistent with the lesions induced following the persistent elevation of intraocular pressure as a result of insufficient drainage of aqueous humor. Thus, the present cases observed in the eyes of MG-W gerbils exemplified the anterior segment dysmorphogenesis associated with inadequate neural crest migration or differentiation, leading to subsequent glaucoma.

Purpose This study aimed to evaluate the effect of advanced paternal age on pregnancy outcomes and sperm parameters following intrauterine insemination (IUI). We used IUI data rather than assisted reproductive technology data, which might mask the effects of sperm impairments. Methods We retrospectively analyzed 1576 IUI cycles in women under 40 years old between April 2012 and May 2016 at the National Center for Child Health and Development in Japan. The main outcomes were clinical pregnancy and live birth. Results The mean male age was significantly lower in cycles that resulted in pregnancy compared with those without pregnancy (38.0 vs 39.1 years; P

The four types of chromogenic selective media that are commercially available in Japan were compared for establishing a Japanese standard method for detecting Cronobacter spp. based on ISO/TS 22964:2006. When assessed using 9 standard Cronobacter spp. strains and 29 non-Cronobacter strains, Enterobacter sakazakii isolation agar, ChromocultTM Enterobacter sakazakii agar, CHROMagarTM E. sakazakii, and XM-sakazakii agar demonstrated excellent inclusivity and exclusivity. Using the ISO/TS 22964:2006 method, the recovered numbers of 38 Cronobacter spp. strains, including 29 C. sakazakii isolates obtained from each medium, were equivalent, indicating that there was no significant difference (p > 0.05) among the four types of chromogenic selective media. Thus, we demonstrated that these four chromogenic selective media are suitable alternatives when using the standard method for detecting Cronobacter spp. in Japan, based on the ISO/TS 22964:2006.

The mouse bioassay for diarrhetic shellfish poisoning toxins has been used worldwide. In this study, dinophysistoxin-1 (DTX-1) and okadaic acid (OA) were compared for toxicity. The lethality rate increased and the median survival time decreased in a dose-dependent manner in both DTX-1 and OA. The median lethal dose value was 150.4 µg/kg (95% confidence interval=130.1–171.2 µg/kg) for DTX-1 and 185.6 µg/kg (95% confidence interval=161.2–209.6 µg/kg) for OA. The toxicity equivalent factor 1:1 has been used for OA and DTX-1 in the EU and Japan. Thus, it may be considered that toxicity potential of DTX-1 has remained underestimated as compared to that of OA and DTX-1 might be more toxic than OA.

Deformed osseous structures have been reported as rare causes of extracranial internal carotid artery (ICA) dissection, including the styloid process and the hyoid bone. Here, the authors describe the first known case of symptomatic ICA dissection caused by a giant osteophyte due to atlantoaxial osteoarthritis. The left ICA was fixed at the skull base and at the ICA portion compressed by the osteophyte, and it was highly stretched and injured between the two portions during neck rotation. The patient was successfully treated with ligation of the affected ICA following balloon test occlusion. Atlantoaxial osteoarthritis should be considered in the differential diagnosis of ICA dissection in patients with a severely deformed cervical spine.

Background Cardioembolic stroke (CE) is usually associated with a larger ischemic area leading to higher morbidity and mortality rates. No biomarkers for CE are available, which causes difficulty in differential diagnosis of CE from other subtypes of acute ischemic stroke.We prospectively evaluated consecutive patients with acute ischemic stroke to identify biomarkers that could distinguish between CE and other subtypes of acute ischemic stroke. Etiological diagnoses were identified according to the National Institute of Neurological Disorders and Stroke (NINDS) 111 classification using clinical examinations, computed tomography (CT), magnetic resonance imaging (MRI), cardiac evaluations, and other tests. The biomarkers N-terminal pro-brain natriuretic peptide (NT-pro-BNP), Thrombin-Antithrombin III Complex (TAT), and D-dimer were determined in blood samples collected within 48 hours of onset and compared between groups with and without CE. Non- CE consisted of atherothrombotic brain infarction (ATBI), lacunar infarction (LI), and other stroke subtypes of unknown cause (other).This study included 279 patients diagnosed with acute ischemic stroke. Serum levels of NT-pro- BNP were significantly higher in those with than in those without CE stroke (p0.0001). Analysis of receiver operating characteristics (ROC) curves indicated that an NT-pro-BNP cutoff of 332 pg/mL provided optimal sensitivity (98.3%) and specificity (75.8%) for distinguishing CE from non-CE.Serum levels of NT-pro-BNP may help in diagnosis of CE during the acute phase and thus allow appropriate therapy t6 prevent subsequent cardiogenic stroke.

The four types of chromogenic selective media that are commercially available in Japan were compared for establishing a Japanese standard method for detecting Cronobacter spp. based on ISO/TS 22964:2006. When assessed using 9 standard Cronobacter spp. strains and 29 non-Cronobacter strains, Enterobacter sakazakii isolation agar, Chromocult

Bacterial translocation (BTL) is defined as the passage of viable bacteria from the gastrointestinal tract to the organs. This study was to elucidate the roles of Peyer's patches (PPs) and/or mesenteric lymph nodes (MLNs) in BTL. Alymphoplastic mutant mice and phenotypically normal heterozygous micewere dominantly colonized with streptomycin-resistant Escherichia coli and BTL was examined. In PP- and MLN-competent mice, BTL to MLNs was detected in 100% of mice, but BTL to organs was rare (25%). On the other hand, in PP- and MLN-deficientmice, BTL to organs was detected in 91% of mice. The results clearly indicate that PPs are not the only site for bacterial entry.

Objective The purpose of this study was to examine the 11C-methionine (MET) and 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) findings of central nervous system (CNS) germinoma and the diagnostic utility of these findings. Methods We retrospectively evaluated the cases of 10 patients who were diagnosed with CNS germinoma according to their histopathological or clinical findings. All the patients underwent pretreatment MET and/or FDG-PET scans, and the resultant images were assessed qualitatively and quantitatively. In the qualitative assessments, we used 3- and 5-grade visual scoring systems for the MET- and FDG-PET images, respectively. In the quantitative assessments, the maximal standardized uptake value (SUVmax) and the ratio of the SUVmax of the tumor (T) divided by the mean SUV for the normal white or gray matter [T/N (WM), T/N (GM)], was calculated. Results The mean and SD values of SUVmax, T/N (WM), and T/N (GM) were 1.9 ± 1.4, 2.5 ± 1.3, and 1.7 ± 0.9 on MET-PET and 5.8 ± 2.2, 1.6 ± 0.5, and 0.8 ± 0.2 on FDG-PET, respectively. On MET-PET, only one lesion was not detected. On the other hand, on FDG-PET all of the lesions exhibited uptake values that were intermediate between those of the normal white matter and gray matter. Conclusion In terms of its tumor-contouring ability, MET is a good tracer for diagnosing CNS germinomas; therefore, MET-PET is considered to be useful for planning biopsies or surgery. Although FDG-PET is capable of detecting CNS germinomas, it produced insufficient image contrast in the present study. Further studies are needed before FDG-PET can be used in clinical examinations of CNS germinoma.

For the surveillance of the prevalence of Campylobacter jejuni and Campylobacter coli in raw chicken products in Japan, a qualitative method, National Institute of Health Sciences Japan (NIHSJ)-02, was developed as an alternative to International Organization for Standardization (ISO) 10272-1:2006. In the NIHSJ-02 culture method, the enrichment step is carried out in a reduced volume of Preston broth at 42 ± 1°C to reduce cost and space, and to prevent the overgrowth of background bacteria. To evaluate the performance of NIHSJ-02, a collaborative study was conducted, and the results obtained by NIHSJ-02 were compared with those obtained using the reference method, ISO 10272-1:2006. Fifteen laboratories participated; each examined 48 minced chicken samples consisting of test samples uninoculated, inoculated with C. jejuni at a low or high level, and inoculated with C. coli at a low level. The average probabilities of detection by NIHSJ-02 across laboratories were 0.033, 0.222, 0.678, and 0.267 in samples uninoculated, inoculated with C. jejuni at a low and high level, and with C. coli at a low level, respectively. Those by ISO 10272-1:2006 were 0.051, 0.128, 0.551, and 0.090. Significantly higher probabilities of detection were determined by NIHSJ-02 compared to ISO 10272-1:2006, except for uninoculated samples. On the other hand, significantly lower frequency of occurrence of background bacteria was observed by NIHSJ-02 (43.1%) compared with ISO 10272-1:2006 (92.6%). NIHSJ-02 showed better performance than ISO 10272-1:2006 with regard to the selective detection of C. jejuni and C. coli in chicken.

The ability of L. monocytogenes to grow in a series of Japanese ready-to-eat (RTE) foods, including boiled baby sardine and Japanese pickle, was tested at two different refrigeration temperatures. In RTE foods in which L. monocytogenes can grow, growth was significantly higher at 10°C than that at 4°C during their shelf lives and growth patterns varied extensively among the different types of foods. However, growth did not occur at 4°C within the shelf life of certain RTE foods, such as broiled squid. The patterns of growth were varied extensively with different sample types. These results suggest that some types of traditional Japanese RTE foods stored at 10°C may be potential sources of listeriosis. To reduce the risk of food-borne listeriosis, studies to determine the contamination levels in RTE foods and the effects of storage temperature on their shelf lives are needed.

The purpose of the study was to compare observer performance in the detection of cerebral infarction on a brain CT using medical-grade liquid crystal display (LCD) monitors calibrated with the gray-scale standard display function and with γ 2.2 and using an iPad with a simulated screen setting.We amassed 97 sample sets, from 47 patients with proven cerebral infarction and 50 healthy control subjects. Nine radiologists independently assessed brain CT on a gray-scale standard display function LCD, a γ 2.2 LCD, and an iPad in random order over 4-week intervals. Receiver operating characteristic (ROC) analysis was performed by using the continuous scale, and the area under the ROC curve (A(z)) was calculated for each monitor.The A(z) values for gray-scale standard display function LCD, γ 2.2 LCD, and iPad were 0.875, 0.884, and 0.839, respectively. The difference among the three monitors was very small. There was no significant difference between gray-scale standard display function LCD and γ 2.2 LCD. However, the A(z) value was statistically significantly smaller for the iPad than the γ 2.2 LCD (p0.05).Observer performance for detecting cerebral infarction on the LCD with γ 2.2 calibration was found to be similar to the LCD with gray-scale standard display function calibration. Although observer performance using the iPad was poorer than that using the other LCDs, the difference was small. Therefore, the iPad could not substitute for other LCD monitors. However, owing to the promising potential advantages of tablet PCs, such as portability, further examination is needed into the clinical use of tablet PCs.

Objective Prediction of motor function after intracerebral hemorrhage (ICH) often poses a diagnostic challenge. This study was performed to investigate whether intraoperative monitoring of motor-evoked potentials (MEPs) could predict postoperative motor function recovery. Methods We reviewed 16 consecutive patients undergoing evacuation of supratentorial ICH with hemiplegia between June 2011 and October 2014. Patients were categorized according to the results of MEPs before and after evacuation of hematoma. The correlation between detection of MEPs and prognosis of motor function was analyzed. Results In 10 of 16 cases (62%), stable MEPs were detected before and after evacuation of hematoma, and postoperative motor function was improved in all cases, including 3 cases with severe preoperative motor impairment on manual muscle test (1–2). In 3 cases (19%) in which MEPs were not detected throughout the procedure, motor function was not improved. In the other 3 cases (19%), MEPs were not measured before evacuation of ICH but were detected after evacuation despite poor prognosis of motor function. The results of postevacuation MEPs were considered false-negative results. Predictions using pre-evacuation MEP results were completely consistent with prognosis for recovery, whereas MEPs obtained during and after evaluation were useful for monitoring. Conclusions Intraoperative MEPs may indicate preservation of pyramidal tracts, and pre-evacuation MEPs can predict motor function outcome after ICH surgery.

Listeria monocytogenes is a foodborne pathogen that causes gastroenteritis, maternofetal infections and meningoencephalitis in humans. Here we report that an intrahost genome mutation alters bacterial acid resistance and the abilities for replication/invasion in tissue cell culture. Among the L. monocytogenes isolates from the recent outbreak in Japan, we found that one food strain, 668, exhibited the greatest acid resistance, whereas one human clinical strain, 690, sharing identical pulsed-field gel electrophoresis (PFGE) and ribotyping patterns, exhibited an acid-sensitive phenotype. Passage of the 668 food strain through the mouse intestine increased its acid sensitivity without altering the macrogenotypes, indicating intrahost alteration of the bacterial acid-resistant phenotype. Genetic and proteomic analyses revealed a link between acid resistance and SigB (RNA polymerase SigmaB subunit) activity. Compared with the strain 668, the clinical and 4 of 5 mice-passaged strains showed a mutation in the rsbW locus, whose product controls the regulation of SigB activity. Corresponding to the SigB activity, the host-passaged strains had reduced abilities to survive inside macrophages and to invade Caco-2 cells, compared with the food strain 668. Overall, we have demonstrated the first example of a host environment promoting the alteration of SigB-dependent acid resistance and host cell-associated actions of L. monocytogenes. Our study provides new insight into the potential role of intrahost environment in the process of bacterial evolution.

The antimicrobial susceptibility of 201 Listeria monocytogenes isolates from foods, environments, animals and human patients in Japan was determined. All isolates were susceptible to ampicillin, the first choice of drug for listeriosis treatment, chloramphenicol, dihydrostreptomycin, erythromycin, enrofloxacin, gentamicin, kanamycin, lincomycin, nosiheptide, salinomycin, vancomycin, and virginiamycin. A human strain was resistant to oxytetracycline. The Minimum Inhibitory Concentration (MIC) for 50% of the strains and the MIC for 90% of the strains were comparable in all the isolates. This is the first investigation to compare antibiotic resistances between isolates from foods and isolates from human patients in Japan. The result showed that most of the isolates were susceptible to antibiotics used in this study.

当研究の目的は，急性脳梗塞の転帰を検討し，同一施設内で急性期から回復期リハビリテーション(以下リハ)を行う病院完結型施設での脳卒中治療の成果と問題点を検証することである．対象は，最近3年間の連続した急性脳梗塞入院患者456例で，ラクナ梗塞170例，アテローム血栓性梗塞151例，心原性脳塞栓症116例，その他梗塞19例に病型分類した．転帰は退院時modified Rankin Scale (mRS)と在宅復帰率により評価した．急性脳梗塞の病型別転帰は，ラクナ梗塞，アテローム血栓性梗塞，心原性脳塞栓症の3病型間で，mRS0–2率(転帰良好群)，mortalityおよび在宅復帰率の全てにおいて有意差を認め，入院時病型診断は転帰予測上重要であった．入院時重症度評価としてのNIH Stroke Scale (NIHSS)は，mRS0–2率および在宅復帰率における有意差を認め，退院時転帰予測に有用であった．急性期治療終了時Functional Independence Measure (FIM) score 90以上例の98.6%が在宅復帰し，FIM score 89以下例の在宅復帰例では回復期リハFIM獲得(FIM gain)値が有意に高かったことから，FIMとFIM gain値は，回復期リハ適応判定および在宅復帰を含めた転帰の指標となった．回復期リハ実施群132例の在宅復帰率は76.5%で，非実施群と比較してmRS3および4例の在宅復帰率が有意に高く，回復期リハは在宅復帰率の向上に役立っていた．急性脳梗塞に対する病院完結型施設での脳卒中治療は，急性期転帰予測，急性期終了時治療評価と回復期リハ適応判定基準および在宅復帰を含めた転帰につき一定の基準で検討することで，急性期から回復期に至る継ぎ目のないリハを含めた計画的な治療が可能となり，在宅復帰率の向上を含め良好な転帰を期待できる．

A 21-year-old man presented with a hemorrhagic central neurocytoma manifesting as acute onset of disturbance of consciousness and right hemiparesis. Computed tomography (CT) demonstrated a tumor in the left lateral ventricle during the course of evaluation for mental alteration 12 days before onset, but the tumor was left untreated because the patient refused to visit a neurosurgical institution. CT on admission revealed a large mass lesion located in the body of the lateral ventricle associated with massive intratumoral and intraventricular hemorrhage. He underwent emergent surgery for evacuation of the tumor with hematoma, and his neurological symptoms gradually recovered after surgery. The present case highlights the possibility of rapid deterioration of symptoms by massive hemorrhage from central neurocytoma. Surgical intervention should thoroughly be considered, if intratumoral hemorrhage is present, as hemorrhage from the central neurocytoma may lead to serious neurological complications.

Listeria monocytogenes, the causative agent of listeriosis, has strong osmotolerance and is able to grow in severe circumstances. Many studies of the mechanisms of listerial osmotolerance have been performed. However, there is much which remains unknown. In previous studies we constructed two kinds of mutant in L. monocytogenes EGD strain to analyse the mechanisms of osmotolerance in L. monocytogenes by molecular genetic methods. In this paper, we summarized the genetical studies of osmotolerance in this bacterium by many researchers and ourselves. First, a transposon-insertional mutant strain was constructed that showed reduced growth in high osmotic agar compared with the parental strain. The results of cloning and sequencing analysis showed that the rel gene, which encodes guanosine tetra- and pentaphosphate synthesis and hydrolysis protein, is involved in osmotolerance in L. monocytogenes. Next, the expression levels of five sigma factor coding genes in L. monocytogenes were examined using real-time polymerase chain reaction (PCR) and it was found that the rpoN gene (the alternative sigma factor RpoN (sigma54)-encoding gene) was activated under high osmotic conditions. A deletion mutant of rpoN was constructed and its response to osmotic stress was analysed. In minimal medium with NaCl and carnitine, an osmoprotectant, the mutant showed deficient growth to that of the parental strain when the starting optical density was high, though the expression level of carnitine transporter operon, opuC, and the rate of carnitine uptake in the mutant was similar to that of EGD. These results suggest that the rpoN mutant may need larger amounts of carnitine which might be needed for its growth under high osmolarity. Through the analysis of these mutants, new insights have been obtained into osmotolerance in L. monocytogenes.

Campylobacter is one of the most frequently diagnosed bacterial causes of human gastroenteritis in Japan and throughout the world. Resistance to quinolones in Campylobacter jejuni and C. coli isolated from humans has emerged in many countries during the past 15 years because fluoroquinolones are the drug of choice for the treatment of suspected bacterial gastroenteritis. Food contaminated with Campylobacter is the usual source of human infection; therefore, the presence of antimicrobial resistance strains in the food chain has raised concerns that the treatment of human infections will be compromised. The use of antimicrobial agents for food animals and in veterinary medicine is suspected to be correlated with an increase in quinolone-resistant strains of Campylobacter in food animals, especially in poultry products. In contrast to macrolide resistance in C. jejuni and C. coli isolated from humans showing a stable low rate, resistant Campylobacter spp. to quinolones have emerged in Japan. The paper summarizes food-borne Campylobacter infection in Japan, and the prevalence and trends of antimicrobial resistance of Campylobacter from the authors' data and other Japanese papers which reported the antimicrobial resistance of Campylobacter.

We clarified collaborations between public health nurses (PHNs) and the private sector, such as nonprofit organizations.Semi-structured interviews were conducted with 11 private sector organizations and 13 PHNs who collaborate with them between December 2012 to October 2013. Interview guides were: overall suicide preventive measurements, details of collaboration between private sector organizations and PHNs, and suicide prevention outcomes/issues. Data from private sector organizations and PHNs were separately analyzed and categories created using qualitative and inductive design. Private sector organizations' and PHNs' categories were compared and separated into core categories by similarities.Six categories were created: 1. establishing a base of mutual understanding; 2. raising public awareness of each aim/characteristic; 3. competently helping high suicidal risk persons detected during each activity; 4. guarding lives and rehabilitating livelihoods after intervention; 5. restoring suicide attempters/bereaved met in each activity; and 6. continuing/expanding activities with reciprocal cohesion/evaluation.PHNs are required to have the following suicide prevention tasks when collaborating with private sector organizations: understanding the private sector civilization, sharing PHN experiences, improving social determinants of health, meeting basic needs, supporting foundation/difficulties each other (Dear editor. Thank you for kind comments. I was going to explain that PHNs and NPOs support each other their foundation of activity and difficulties in their activities. The foundations include knowledge, information, budgets, manpower etc. The difficulties mean like suffering faced with suicide during activities.), and enhancing local governments' flexibilities/ promptness.

Listeriosis is a relatively rare foodborne illness but can be life threatening with high fatality rates. In Japan, the incidence of listeriosis has been very low for the past 40 years compared with that of Western Europe and North America. We hypothesized that less Listeria monocytogenes contamination in Japanese foods would be related to the lower incidence in Japan. For this purpose, we collected data of Japanese foods contaminated with L. monocytogenes, mainly from Japanese-written reports, and reviewed them. From this review, we found that the proportion of L. monocytogenes, Listeria spp. isolation from foods in Japan is similar to those reported from other countries and that other factors might be responsible for the lower occurrence of listeriosis.

Quantitative analyses of Listeria monocytogenes in imported ready-to-eat (RTE) foods sold at retail stores in Japan were performed. Of the 77 non-cooked meat products, 6 samples (7.8%) tested positive. The levels of contamination of 4 of the samples were below 100 colony-forming units (CFU)/g, which is the microbiological criterion for L. monocytogenes in RTE foods as determined by Codex. However, Listeria cells at levels of 100 and 400 CFU/g were detected in a salami sample and a raw ham sample, respectively. All of the 70 cheese samples and the 3 samples made from raw ham and cheese showed negative test results. These results suggest that imported RTE foods are potential sources of the causative agent of listeriosis.

Bacterial translocation (BTL) is defined as the passage of viable bacteria from the gastrointestinal tract to the organs. This study was to elucidate the roles of Peyer's patches (PPs) and/or mesenteric lymph nodes (MLNs) in BTL. Alymphoplastic mutant mice and phenotypically normal heterozygous micewere dominantly colonized with streptomycin-resistant Escherichia coli and BTL was examined. In PP- and MLN-competent mice, BTL to MLNs was detected in 100% of mice, but BTL to organs was rare (25%). On the other hand, in PP- and MLN-deficientmice, BTL to organs was detected in 91% of mice. The results clearly indicate that PPs are not the only site for bacterial entry.

The activity of prokaryotic and mammalian thymine glycol (Tg) glycosylases including Escherichia coliendonuclease III (Endo III) and endonuclease VIII (Endo VIII) and mouse Endo III homologue (mNth1) for formamidopyrimidine (Fapy) has been investigated using defined oligonucleotide substrates. 2,6-Diamino-4-hydroxy-5-N-methylformamidopyrimidine, a methylated Fapy derived from guanine, was site specifically incorporated in the oligonucleotide. The substrates containing Fapy:N pairs (N = A, G, C, T) as well as a Tg:A pair, a physiological substrate of Endo III, Endo VIII, and mNth1, were treated by the enzymes and nicked products were quantified by gel electrophoresis. The activity of Endo III and Endo VIII for Fapy varied markedly depending on the paired base, being the highest with G (activity relative to Tg = 0.55 (Endo III) and 0.41 (Endo VIII)) and the lowest with C (0.05 (Endo III) and 0.06 (Endo VIII)). In contrast, mNth1 recognized all Fapy pairs equally well and the activity was comparable to Tg. The results obtained in the nicking assay were further substantiated by the analysis of the Schiff base intermediate using NaBH4trapping assays. These results indicate that Escherichia coli and mammalian Tg glycosylases have a potential activity to recognize Fapy. However, as demonstrated for Fapy:C pairs, their distinctive activities implicate unequal participation in the repair of Fapy lesions in cells.

SummaryIn March 2012, two patients were transported urgently to the hospital in Tottori Prefecture, Japan, because of symptoms suggestive of botulism. Botulinum neurotoxin type A was detected in the clinical specimens and the food consumed by the two patients (vacuum packed adzuki-batto, a sweet adzuki bean soup containing noodles). We were able to make a prompt diagnosis of food botulism associated with the consumption of adzuki-batto, from which the causative pathogen Clostridium botulinum Ab was cultured.

Appropriate handling and controlled temperature prevent cross-contamination and proliferation of contaminants in foods, thereby reducing the incidences of food-borne gastroenteritis in Japan. However, the incidence of Campylobacter jejuni/coli infection did not markedly decrease and has become one of the major causes of food-borne diseases. C. jejuni and C. coli are widespread in warm-blooded domestic animals; therefore, food products may easily become contaminated during processing. C. jejuni and C. coli do not proliferate in foods, nor resistant to freezing, drying and oxidative stresses, and the number is greatly reduced under such conditions. These properties should be considered for risk management of Campylobacter in food processing and manufacturing.

Purpose The aim of our study was to examine the usefulness of PET with C-methionine (MET) and F-fluorodeoxyglucose (FDG) in the differentiation of glioblastoma multiforme (GBM) and intracranial diffuse large B-cell lymphoma (DLBCL). Materials and methods We evaluated 22 patients retrospectively with an enhancing brain tumor on MRI, including 15 GBM and 7 DLBCL, which was confirmed by histopathology. Dynamic PET scans with MET and FDG were performed for preoperative differential diagnosis. We assessed the images qualitatively and quantitatively. In quantitative assessment, the SUVmax was used on FDG PET and both late and early phases on MET PET. In addition, the ratio of SUVmax in the late and early phases on MET-PET was evaluated (ΔSUVmax). Results SUVmax on FDG PET of DLBCL was significantly higher than that of GBM. Setting an SUVmax of 12.0 as the cutoff for differentiating DLBCL from GBM, 1 GBM and 1 DLBCL were found to be false-positive and false-negative, respectively.SUVmax in the late and early phases of MET-PET was not significantly different between DLBCL and GBM; however, we also found significant differences in ΔSUVmax on MET-PET. Using ΔSUVmax 1.17 as the cutoff, we could differentiate DLBCL from GBM completely. In the present study, ΔSUVmax on MET-PET was slightly superior to SUVmax on FDG PET. Conclusions Both SUVmax on FDG PET and ΔSUVmax on MET-PET were considered to be good diagnostic tests when encountering difficulties in this differential diagnosis.

Quantitative analyses of Listeria monocytogenes in imported ready-to-eat (RTE) foods sold at retail stores in Japan were performed. Of the 77 non-cooked meat products, 6 samples (7.8%) tested positive. The levels of contamination of 4 of the samples were below 100 colony-forming units (CFU)/g, which is the microbiological criterion for L. monocytogenes in RTE foods as determined by Codex. However, Listeria cells at levels of 100 and 400 CFU/g were detected in a salami sample and a raw ham sample, respectively. All of the 70 cheese samples and the 3 samples made from raw ham and cheese showed negative test results. These results suggest that imported RTE foods are potential sources of the causative agent of listeriosis.

The sequence of neural progenitor cell (NPC) damage induced in fetal rat brain by transplacental exposure to busulfan, an antineoplastic bifunctional-alkylating agent, on gestational day 13 was examined by immunohistochemical and real-time RT-PCR analyses. Following busulfan treatment, pyknotic NPCs first appeared in the medial layer and then extended to the dorsal layer of the ventricular zone (VZ) of the telencephalon. Pyknotic NPCs that were immunohistochemically positive for cleaved caspase-3, i.e. apoptotic NPCs, began to increase at 24 h after treatment, peaked at 48 h, and returned to the control levels at 96 h. On the other hand, the index (%) of phospho-histone H3-positive NPCs, i.e. mitotic NPCs, and that of BrdU-positive NPCs, i.e. S-phase cells, decreased in accordance with the increase in the index of apoptotic NPCs. Prior to the peak time of apoptotic NPCs, the indices of p53- and p21-positive NPCs peaked at 36 h. In addition, the expression levels of p21 and Puma (p53-target genes) mRNAs were elevated in real-time RT-PCR analysis. These findings indicated that busulfan not only induced apoptosis through the p53-mediated intrinsic pathway but also inhibited cell proliferation in NPCs, resulting in a reduction of the width of the telencephalon. On the other hand, in spite of up-regulation of p21 expression, the expression of cyclin D1, part of the cell cycle machinery of the G1/S transition, and the expression levels of Cdc20 and cyclin B1 which are involved in G2/M transition, showed no changes, giving no possible information of busulfan-induced cell cycle arrest in NPCs.

The relationship between cyclooctadepsipeptides and their anthelmintic efficacy was examined by converting the natural products, PF1022A, PF1022E and PF1022H. Some analogues substituted at the para position of the phenyllactate moiety showed higher or equivalent activity against the parasitic nematode, Ascaridia galli in chicken when compared with the parent compounds. It is suggested that lipophilicity and the polar surface area, in addition to structural requirements of the derivatives, influenced the anthelmintic efficacy in vivo.

Johne disease is ruminant chronic granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP). The domestic animals infected with this pathogen present severe weight loss due to chronic diarrhea and a reduction in lactation yield. These result in enormous economic loss since the affected animals are subsequently subject to artificial selections and disinfection of the environment are absolutely necessary. Furthermore, MAP has been suspected to have pathological relationship to Crohn's disease, human chronic granulomatous enteritis. The bacterium grows slower on solid culture and its colony becomes visible after two months of culture. In Japan, there has been almost no investigation on pasteurization temperature of commercial milk using MAP. It comes from the fact that the growth rate of MAP is very slow and that MAP is a related species to Mycobacterium tuberculosis, which pasteurization condition has been well defined. The studies on the pasteurization conditions of commercial milk have been mainly targeted to reduce the risk of infection to Coxiella and Mycobacterium tuberculosis. However, there has been a concern about the possibility that MAP is remained in pasteurized milk because MAPs form an aggregate and the bacterium at its center may not receive enough heat to get pasteurized. From these reasons, the present study aims to investigate validity of the current pasteurization conditions of commercial milk by implementing experimental pasteurization at various pasteurization temperatures using milk experimentally infected with MAP, and to clarify if MAP is eliminated at these temperatures in order to achieve smooth enforcement of the current ministry order. We conducted plant pasteurization experiment at four pasteurization conditions (high temperature, short time (HTST); 82, 77, 72 degrees C for 15 seconds and low temperature, long time (LTLT); 63 degrees C for 30 minutes) using two MAP strains, ATCC19698 and OKY-20. In conclusion, there appeared no colony of the two MAP strains formed from the milk pasteurized at the four pasteurization conditions examined.

A 55-year-old man presented with intracerebral hemorrhage (ICH) without subarachnoid hemorrhage (SAH) manifesting as acute onset of consciousness disturbance and right hemiparesis. Computed tomography showed ICH mainly localized in the left putamen, but no evidence of SAH. Magnetic resonance angiography demonstrated a cerebral aneurysm originating from the bifurcation of the left internal carotid artery, which was considered to be responsible for the ICH. The patient underwent emergent intravascular surgery for coil embolization of the aneurysm, and his neurological symptoms gradually recovered with rehabilitation after surgery. Although ICH without SAH is a rare presentation of cerebral aneurysm, ruptured cerebral aneurysm should be considered as a potential cause of ICH. The localization and extent of ICH may be suggestive of latent cerebral aneurysm in such cases.

A 21-year-old man presented with a hemorrhagic central neurocytoma manifesting as acute onset of disturbance of consciousness and right hemiparesis. Computed tomography (CT) demonstrated a tumor in the left lateral ventricle during the course of evaluation for mental alteration 12 days before onset, but the tumor was left untreated because the patient refused to visit a neurosurgical institution. CT on admission revealed a large mass lesion located in the body of the lateral ventricle associated with massive intratumoral and intraventricular hemorrhage. He underwent emergent surgery for evacuation of the tumor with hematoma, and his neurological symptoms gradually recovered after surgery. The present case highlights the possibility of rapid deterioration of symptoms by massive hemorrhage from central neurocytoma. Surgical intervention should thoroughly be considered, if intratumoral hemorrhage is present, as hemorrhage from the central neurocytoma may lead to serious neurological complications.

SDS-PAGE showed that human salivary alpha-amylase family A (HSA-A) was converted to family B (HSA-B) in human saliva. This conversion did not occur in the supernatant of saliva which had been centrifuged at 105,000 x g for 60 min. An enzyme which catalyzed the conversion existed in the insoluble fraction of human saliva. The enzyme was solubilized with nonionic or zwitterionic detergents, and showed the maximum activity around pH 6. It was stable between pH 4 and 10, and at a temperature lower than 40 degrees C. The enzyme reduced the molecular weight of HSA-A (62,000) to the same molecular weight (58,000) as that of HSA-B without forming any intermediate. It also changed the PAGE pattern of multiple forms of HSA-A to the same pattern as that of HSA-B. It was not inhibited by protease inhibitors, and it did not destroy the reactivity of HSA-A with anti-human salivary alpha-amylase antiserum. The enzyme diminished the reactivity of HSA-A with concanavalin A. These results indicate that HSA-A was converted to HSA-B through the release of sugar chains by the action of the enzyme in the insoluble fraction of human saliva.

Vibrio cholerae is an aetiological agent of cholera that inhabits marine and estuarine environments. It can survive harsh environments by entering the viable but non-culturable (VBNC) state, but the related changes in gene expression have not been described. Here, we experimentally induced the VBNC state in V. cholerae O1, by incubation in artificial seawater at 4 degrees C. Bacterial cells that were incubated for 70 days retained their membrane integrity and were pathogenic, colonizing the gut of iron-dextran-treated mice, even though they formed no colonies on tryptic soy agar (TSA) or TSA amended with pyruvate. We therefore used this stage of cells as the VBNC bacteria. We compared the global transcription pattern of the VBNC cells with that of stationary-phase cells grown in rich medium. A total of 100 genes were induced by more than fivefold in the VBNC state, and the modulated genes were mostly those responsible for cellular processes. Furthermore, real-time RT-PCR analysis verified the changes in the expression levels, showing that the VC0230 [iron(III) ABC transporter], VC1212 (polB), VC2132 (fliG) and VC2187 (flaC) mRNAs were increased in the non-culturable state. Thus, these genes may be suitable markers for the detection of VBNC V. cholerae. To our knowledge, this is the first report of a comprehensive transcriptome analysis of V. cholerae in the VBNC state. The significance of this gene expression profile compared with those of in vivo isolates and non-stressed bacteria (culturable in vitro) is its potential to provide information about the public health risk from dormant bacteria.