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A 67-year-old woman with a 2-day history of a fever, headache and disturbed consciousness was admitted to our hospital. Bacillus subtilis was isolated from both the cerebrospinal fluid and blood. She was cured by the administration of vancomycin. Next-generation sequencing identified the strain as B. subtilis var. natto, the same strain found in natto, which this patient ate daily. We suspected that some of the B. subtilis that caused the infection may have actually been B. subtilis var. natto.

With the emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), a high-speed and convenient detection technology should be at the forefront of medical care worldwide. This study evaluated the usefulness of GeneSoC, a compact, high-speed reciprocal flow quantitative reverse transcription polymerase chain reaction system, for the detection of SARS-CoV-2. The results support the use of this system for the rapid identification of SARS-CoV-2. This approach can contribute to the strategic selection of initial management strategies for patients with COVID-19.

Background Capnocytophaga canimorsus is a gram-negative bacterium and an oral commensal in dogs and cats, but occasionally causes serious infections in humans. Septicemia is one of the most fulminant forms, but diagnosis of C. canimorsus infection is often difficult mainly because of its very slow growth. C. canimorsus infective endocarditis (IE) is rare and is poorly understood. Since quite a few strains produce β-lactamase, antimicrobial susceptibility is pivotal information for adequate treatment. We herein report a case with C. canimorsus IE and the results of drug susceptibility test. Case presentation A 46-year-old man had a dog bite in his left hand 3 months previously. The patient was referred to our hospital for fever (body temperature > 38 °C), visual disturbance, and dyspnea. Echocardiography showed aortic valve regurgitation and vegetation on the leaflets. IE was diagnosed, and we initially administered cefazolin and gentamycin assuming frequently encountered microorganisms and the patient underwent aortic valve replacement. C. canimorsus was detected in the aortic valve lesion and blood cultures. It was also identified by 16S ribosome DNA sequencing. Ceftriaxone were started and continued because disk diffusion test revealed the isolate was negative for β-lactamase and this case had cerebral symptoms. The patient successfully completed antibiotic treatment following surgery. Conclusions We diagnosed C. canimorsus sepsis and IE by extended-period blood cultures and 16S ribosome DNA sequencing by polymerase chain reaction, and successfully identified its drug susceptibility.

To assess the time trend of diagnostic accuracy of pre- and post-eradicationA series 100 of conventional endoscopic image sets taken from subjects undergoing gastric cancer screening at a polyclinic were evaluated by 5 experienced assessors. Each assessor independently examined endoscopic findings according to the Kyoto classification and then determined theThe overall diagnostic accuracy ofDiagnostic accuracy of

Purpose . The new third-generation sequencing platform MinION is an attractive maintenance-free and disposable portable tool that can perform long-read and real-time sequencing. In this study, we validated this technology for the identification of pathogens from positive blood culture (BC) bottles. Methodology . A total of 38 positive BC bottles were collected from patients with bloodstream infections, and 18 isolates of Gram-negative (GN) bacteria and 20 isolates of Gram-positive (GP) bacteria were identified from these using 16S rRNA sequencing and then used in this study. DNA was extracted from each aliquot using an extraction protocol that combined glass bead beating and chemical lysis. Up to 200 ng of each purified DNA sample was processed for library preparation and whole-genome sequencing was performed on up to 12 samples through a single MinION flow cell. Results . All GN bacteria identifications made by MinION sequencing for 30 min using the What’s In My Pot? (WIMP) workflow via EPI2ME on the basis of the most frequent classified reads were consistent with those made by 16S rRNA sequencing. On the other hand, for GP bacteria specimens, the identification results for 16S rRNA sequencing and MinION were only in agreement in 12 out of 20 (60.0 %) cases. ARMA analysis was able to detect extended-spectrum β-lactamase (ESBL)-associated genes among various antimicrobial resistance-related genes. Conclusion . We demonstrated the potential of the MinION sequencer for the identification of GN bacteria from positive BC bottles and the confirmation of an ESBL phenotype. This innovative sequence technology and its application could lead to a breakthrough in the diagnosis of infectious diseases.

Purpose. Bloodstream infections are major causes of morbidity and mortality that lead to prolonged hospital stays and higher medical costs. In this study, we aimed to evaluate the MinION nanopore sequencer for the identification of the most dominant pathogens in positive blood culture bottles. Methodology. 16S and ITS1-5.8S-ITS2 rRNA genes were amplified by PCR reactions with barcoded primers using nine clinical isolates obtained from positive blood bottles and 11 type strains, including five types of Candida species. Barcoded amplicons were mixed, and multiplex sequencing with the MinION sequencer was performed. In addition, barcoded PCR amplicons were sequenced by Sanger sequencing to validate the performance of the MinION. Results. The bacterial and Candida spp. identified by MinION sequencing, based on the highest homology of reference sequences from the NCBI gene databases, agreed with the matrix-assisted laser desorption ionization time of flight mass spectrometry results, excepting the closely related species Streptococcusand Escherichia coli. The ‘pass’ reads obtained within about 10 min of sequencing were sufficient to identify the pathogens. The average values of sequence identities with 1D2 chemistry and the R9.5 flow cell were around 99 %; thus, frequent sequence errors did not affect species identification based on amplicon sequencing. Conclusion. We have established a rapid, portable and economical technique for the identification of pathogens in positive blood culture bottles through a novel MinION nanopore sequencer amplicon sequencing scheme, which replaces traditional Sanger sequencing.

Background The major symptoms of irritable bowel syndrome (IBS) are changes in bowel habits and abdominal pain. Psychological stress is the major pathophysiological components of IBS. Corticotropin-releasing factor (CRF) is a well-known integrator in response to psychological stress. In this study, a novel CRF1 receptor antagonist T-3047928 was evaluated in stress-induced IBS models of rats to explore its potency for IBS. Methods Plasma adrenocorticotropic hormone (ACTH) levels after intravenous oCRH challenge were measured as a pharmacodynamic marker. Efficacies of oral T-3047928 were compared with oral alosetron, a 5-HT3 antagonist, on conditioning fear stress (CFS)-induced defecation, restraint stress (RS)-induced acute visceral pain, specific alteration of rhythm in temperature (SART) stress-induced chronic visceral pain, and normal defecation. Results T-3047928 (1-10 mg/kg, p.o.) demonstrated a dose-dependent inhibition on oCRH-induced ACTH secretion. In disease models, T-3047928 suppressed fecal pellet output induced by CFS and improved both acute and chronic visceral hypersensitivity induced by RS and SART stress, respectively. Alosetron was also efficacious in stress-induced defecation and visceral pain models at 1 and 10 mg/kg, respectively. Alosetron, however, also suppressed normal defecation at lower those. On the other hand, T-3047928 did not change normal defecation even at higher dose than those in disease models. Conclusion T-3047928 is an orally active CRF1 antagonist that demonstrated potent inhibitory effects in stress-associated IBS models with no effect on normal defecation. Therefore, it is suggested that T-3047928 may have a potency as a novel option for IBS-D therapy with minimal constipation risk.

Background Klebsiella variicola and K. quasipneumoniae are new species distinguishable from K. pneumoniae but they are often misidentified as K. pneumoniae in clinical settings. Several reports have demonstrated the possibility that the virulence factors and clinical features differ among these three phylogroups. In this study, we aimed to clarify whether there were differences in clinical and bacterial features between the three phylogroups isolated from patients with bloodstream infections (BSIs) in Japan. Methods Isolates from all patients with BSIs caused by K. pneumoniae admitted to two hospitals between 2014 and 2017 (n = 119) were included in the study. Bacterial species were identified via sequence analysis, and their virulence factors and serotypes were analyzed via multiplex PCR results. Clinical data were retrieved from medical records. Results Of the 119 isolates, 21 (17.7%) were identified as K. variicola and 11 (9.2%) as K. quasipneumoniae; K1 serotype was found in 16 (13.4%), and K2 serotype in 13 (10.9%). Significant differences in the prevalence of rmpA, iutA, ybtS, entB and kfu (p allS genes (p K. variicola was more frequently detected in patients over 80 years old compared with other Klebsiella species (p K. quasipneumoniae more frequently occurred in patients with malignancy (p Conclusions Our findings demonstrated the differences in bacterial pathogenicity and clinical features among these three phylogroups. Further epidemiological studies into BSI caused by Klebsiella species are warranted.

Background and Aim: Reddish depressed lesions (RDLs) frequently observed in patients following Helicobacter pylori eradication are indistinguishable from gastric cancer. We examined the clinical and histological feature of RDLs and its relevant endoscopic diagnosis including magnifying narrow-band imaging (M-NBI). Methods: We enrolled 301 consecutive patients with H. pylori eradication who underwent endoscopy using white light imaging (WLI). We examined the prevalence and host factors contributing to the presence of RDLs. Next, we used M-NBI in 90 patients (104 RDLs), and compared the diagnostic efficacy between M-NBI and WLI groups using propensity-score matching analysis. Results: In 301 patients after eradication, 117 (39%) showed RDLs. Male, open-type atrophy, and gastric cancer history were risk factors for RDLs. A gastric biopsy was needed in 83 (71%) during WLI observation and only 2 were diagnosed with adenocarcinoma. In M-NBI group, a biopsy was performed in 21 (20%), and 9 were diagnosed with adenocarcinoma. A biopsy was required in fewer patients, and the positive predictive value of a biopsy was statistically higher in M-NBI than in the WLI group (p < 0.01). Conclusions: RDLs are frequently observed in high-risk patients for gastric cancer after eradication. M-NBI demonstrated significantly superior diagnostic efficacy with respect to RDL.

It is clinically important to diagnose drug-induced gastric lesions correctly. Recently, the use of proton pump inhibitors (PPI) has increased worldwide. The histological features induced by PPI have been reported; however, few reports have described endoscopic findings induced by PPI. Therefore, we aimed to clarify the characteristic endoscopic features in PPI users and associated pathogenic factors.We prospectively registered 1007 consecutive participants (70 PPI users and 937 nonusers) who underwent endoscopic examination for cancer screening in three hospitals/clinics. Clinical data and endoscopic findings were recorded in the registration forms. We compared the endoscopic features between the two groups and evaluated contributing factors via univariate and multivariate analyses.Multiple white elevated lesions (MWEL) and cobblestone-like mucosa (CLM) were more commonly observed in PPI users compared with nonusers (p .01). Foveolar hyperplastic polyps were also frequently observed in PPI users but were not statistically significantly different (p = .06). MWEL and CLM were more frequently observed in older patients than in younger patients. MWEL was more frequently observed in female patients than in male patients; however, CLM was predominantly observed in male patients.MWEL and CLM are characteristic endoscopic features in PPI users. A gender-associated difference was noted in terms of the frequency of these lesions.

We evaluated and compared the diagnostic potential of computed tomography (CT) and herniography in the identification of inguinal hernias. Herniography is typically utilized in the diagnosis of inguinal hernias in our institute. We utilized transabdominal preperitoneal repair in the treatment of inguinal hernias since October 2010 and performed a laparoscopic diagnosis followed by single-incision laparoscopic surgery for totally extraperitoneal repair since November 2013. We herein evaluated and compared the diagnostic potential of CT and herniography. From February 2012 to March 2016, 46 patients (male/female: 43/3; median age: 70 years) with 63 inguinal hernias were investigated using CT and herniography. We classified each inguinal hernia according to the Japan Hernia Society classification using CT (prone position) and herniography, and evaluated the accuracy of each of the diagnostic methods. A total of 29 patients had unilateral hernias and 17 had bilateral hernias; 62% (39/63) and 84% (53/63) of the inguinal hernias were correctly diagnosed and classified by CT and herniography, respectively. A total of 87% (55/63) and 98% (62/63) of the inguinal hernias were detected by CT and herniography, respectively. Thus, herniography was found to be more sensitive than CT and the difference in the detectability of inguinal hernias by the 2 methods was statistically significant. Herniography appears to be superior to CT, for both the classification and detection of inguinal hernias.

Fibromyalgia is characterized by chronic widespread musculoskeletal pain. A hypofunction in descending pain inhibitory systems is considered to be involved in the chronic pain of fibromyalgia. We examined functional changes in descending pain inhibitory systems in rats with specific alternation of rhythm in temperature (SART) stress, by measuring the strength of diffuse noxious inhibitory controls (DNIC). Hindpaw withdrawal thresholds to mechanical von Frey filament or fiber-specific electrical stimuli by the Neurometer system were used to measure the pain response. To induce DNIC, capsaicin was injected into the intraplantar of the forepaw. SART-stressed rats were established by exposure to repeated cold stress for 4 days. In the control rats, heterotopic intraplantar capsaicin injection increased withdrawal threshold, indicative of analgesia by DNIC. The strength of DNIC was reduced by naloxone (μ-opioid receptor antagonist, intraperitoneally and intracerebroventricularly), yohimbine (α2-adrenoceptor antagonist, intrathecally), and WAY-100635 (5-HT1A receptor antagonist, intrathecally) in the von Frey test. In SART-stressed rats, capsaicin injection did not increase withdrawal threshold in the von Frey test, indicating deficits in DNIC. In the Neurometer test, deficient DNIC in SART-stressed rats were observed only for Aδ- and C-fibers, but not Aβ-fibers stimulation. Analgesic effect of intracerebroventricular morphine was markedly reduced in SART-stressed rats compared with the control rats. Taken together, in SART-stressed rats, capsaicin-induced DNIC were deficient, and a hypofunction of opioid-mediated central pain modulation system may cause the DNIC deficit.

The in vitro susceptibilities of Bacteroides fragilis to antimicrobial agents, especially to carbapenem, are a major concern in the treatment of patients with bloodstream infections. In this study, 50 isolates of B. fragilis were obtained from positive blood bottles from 2014 to 2019 in Saitama, Japan. Their susceptibility to ampicillin/sulbactam was reduced to 70.0% compared with a previous report, whereas they were still sufficiently susceptible to piperacillin/tazobactam (94.0%). Five cfiA-positive isolates (5/50, 10.0%) were identified that were resistant to doripenem and meropenem, and two of them carried an insertion sequence located upstream of the cfiA-coding region. In particular, imipenem should be considered as a first-line carbapenem for the empirical treatment of B. fragilis infection because only insertion sequence and cfiA double-positive strains showed resistance to imipenem. Thirty-six percent of the isolates had a reduced minimum inhibitory concentration for moxifloxacin. In addition, metronidazole should still be considered as an active agent for B. fragilis because all isolates were susceptible to this antibiotic and the prevalence of the nim gene was low in Japan.

Reddish depressed lesions (RDLs) frequently observed in patients following Helicobacter pylori eradication are indistinguishable from gastric cancer. We examined the clinical and histological feature of RDLs and its relevant endoscopic diagnosis including magnifying narrow-band imaging (M-NBI).We enrolled 301 consecutive patients with H. pylori eradication who underwent endoscopy using white light imaging (WLI). We examined the prevalence and host factors contributing to the presence of RDLs. Next, we used M-NBI in 90 patients (104 RDLs), and compared the diagnostic efficacy between M-NBI and WLI groups using propensity-score matching analysis.In 301 patients after eradication, 117 (39%) showed RDLs. Male, open-type atrophy, and gastric cancer history were risk factors for RDLs. A gastric biopsy was needed in 83 (71%) during WLI observation and only 2 were diagnosed with adenocarcinoma. In M-NBI group, a biopsy was performed in 21 (20%), and 9 were diagnosed with adenocarcinoma. A biopsy was required in fewer patients, and the positive predictive value of a biopsy was statistically higher in M-NBI than in the WLI group (p0.01).RDLs are frequently observed in high-risk patients for gastric cancer after eradication. M-NBI demonstrated significantly superior diagnostic efficacy with respect to RDL.

Background and aims Pathogenic auto-antibodies produced by plasma cells are key drivers of many auto-immune diseases such as Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA), and Sjogren’s Syndrome (SS). In addition, solid organ transplant rejection is also mediated by antibodies produced against the donor organ. Plasma cells are highly metabolically active antibody factories and thus sensitive to depletion by proteasome inhibitors. Ixazomib, an oral proteasome inhibitor, was recently approved in the US and Canada for use in combination with lenalidomide and dexamethasone in patients with multiple myeloma who have received at least 1 prior therapy. The Keyhole limpet hemocyanin (KLH) model of T cell-dependent antigen response was used to determine if ixazomib depletes plasma cells resulting in a reduction of antibodies. Methods Briefly, rats were immunised with KLH and TiterMax adjuvant then treated with ixazomib twice weekly until study termination. Results Treatment with ixazomib significantly inhibited anti-KLH antibodies by 34% (p Conclusions Ixazomib depleted plasma cells resulting in reduced antibodies suggesting further preclinical studies are warranted in diseases with pathogenic antibodies such as SLE, RA, SS and solid organ transplant rejection.

Background and aims Auto-antibodies to nuclear constituents and type I Interferons (IFN) such as IFN-a play key roles in pathogenesis of Systemic Lupus Erythematosus (SLE). Ixazomib, an oral proteasome inhibitor, approved in the US and Canada for use in combination with lenalidomide and dexamethasone in patients with multiple myeloma who have received at least 1 prior therapy. Proteasome inhibitors like ixazomib that may deplete plasma cells and cellular sources of IFN-a are also attractive for autoimmune diseases like SLE. To investigate the potential of ixazomib the MRL/lpr model was used as it has extensively been shown to replicate many features of SLE. Methods MRL/lpr animals received oral ixazomib twice a week for 4 weeks. Results Ixazomib suppressed the time-dependent increase in anti-dsDNA IgG antibodies, resulting in 73% (p Conclusions These findings suggest that ixazomib may be an effective agent for treating antibody-mediated diseases such as SLE by depleting both plasma cells the source of pathogenic antibodies and pDCs the main source of type I IFN production. An ongoing randomised, double-blind phase Ib study is investigating multiple rising doses of Ixazomib (MLN9708) for the treatment of patients with ISN/RPS class III, IV or V lupus nephritis who have not responded adequately to current therapy.

A 63-year-old man visited an emergency outpatient unit with the chief complaints of melena and lightheadedness. At the time of the visit, blood tests showed Hb of 4.3 g/dL, suggesting severe anemia, and he exhibited repeated melena, even after hospitalization. Small intestinal bleeding was suspected during endoscopic examination of the lower gastrointestinal tract, and abdominalCT examination suggested a 3.5 cm tumor-like lesion in the jejunum. He was diagnosed as having bleeding of a tumor in the small intestine and consequently underwent laparoscopic surgery. Based on intraabdominal observation, Meckel 's diverticulum was confirmed in the jejunum, 100 cm from the ileocecal region, along with a 4 cm tumor in the upper jejunum, located 50 cm from Treitz's ligament. The tumor was visually confirmed to be sarcomatoid with no direct invasion to the surrounding tissues and no disseminated node, showing favorable mobility. These lesions were exteriorized from the abdominal cavity for resection and anastomosis, and the surgery was completed with no severe complications. It was diagnosed histopathologically as a gastrointestinal stromal tumor(GIST)in the small intestine, and no postoperative adjunctive chemotherapy was administered because the case was considered low risk based on the tumor diameter and the number of mitosis events. At present, 1 year after the surgery, the patient is under follow-up observation on an outpatient basis with no findings to suggest recurrence or metastasis.

A series of piperazine ureas was designed, synthesized, and evaluated for their potential as novel orally available fatty acid amide hydrolase (FAAH) inhibitors that are therapeutically effective against pain. We carried out an optimization study of the lead compound 3 to improve its DMPK profile as well as in vitro potency. We identified the thiazole compound 60j with potent inhibitory activity, high brain permeability, and good bioavailability. Compound 60j showed a potent and dose-dependent anti-nociceptive effect in the acetic acid-induced writhing test in mice.

Infiltrated macrophages (Mϕ) are believed to cause pathological changes in the surrounding adipocytes through the secretion of active molecules in visceral fat. Matrix metalloproteinase (MMP)-3 is secreted from Mϕ, and enhances expression of the inflammatory cytokines through the activation of toll-like receptor (TLR) 2. Visceral adipocytes express high levels of vascular endothelial growth factor (VEGF), and the degree of visceral fat accumulation is associated with the plasma VEGF concentration in obese subjects. The aim of the study is to clarify the role of MMP-3 in the enhancement of the free fatty acids (FFAs)-induced VEGF expression through TLR2 in visceral adipocytes. One mM FFAs induced VEGF mRNA and protein expression in 3T3-L1 adipocytes. The FFAs-induced VEGF expression was mostly mediated by TLR2. A high fat intake increased the VEGF mRNA expression in visceral fat and the VEGF concentration in plasma, accompanied with the increase in the plasma FFAs concentration in mice. These increases were largely inhibited in TLR2-deficient mice. The FFAs-induced VEGF expression was increased in the presence of Mϕ-conditioned medium (CM) in adipocytes, and the enhancement was inhibited by a MMP-3 inhibitor or a neutralizing antibody against MMP-3. The active form of MMP-3 induced the VEGF mRNA expression, as well as TLR2, in adipocytes. The increase in the VEGF expression by MMP-3 was inhibited by the treatment with siRNA for TLR2. The enhancement of FFAs-induced TLR2 expression by Mϕ-CM was inhibited by blocking of the MMP-3. The increase in the VEGF mRNA expression by Mϕ-CM or MMP-3 was partially inhibited by a neutralizing antibody against TNF-α. These results indicate that MMP-3 in Mϕ-CM enhances the FFAs-induced VEGF expression in adipocytes through the increase in the TLR2 expression. The MMP-3 secreted from the infiltrated Mϕ may be a regulator of the VEGF expression in visceral adipocytes.

Adipocytes accumulated in the visceral area change their function to induce tumor necrosis factor-alpha (TNF-alpha) secretion with concomitant matrix metalloproteinase (MMP)-3 induction in mice. This study was performed to clarify the role of macrophages (Mphi)-secreted MMP on the functional changes in adipocytes using a culture system.Cultures of 3T3-L1 adipocytes with THP-1 Mphi or the Mphi-conditioned medium were used to investigate the role of Mphi-MMP on the TNF-alpha gene in 3T3-L1 adipocytes by the addition of MMP inhibitors. For animal experiments, male C57BL/6J mice were rendered insulin resistant by feeding a high-fat diet, and the expression of an Mphi marker F4/80, and MMP-3 genes in mesenteric and subcutaneous fat tissue specimens were examined.Mphi-conditioned media (Mphi-CM) increased the levels of TNF-alpha mRNA expression in 3T3-L1 adipocytes, and these adipocyte responses were abolished by treatment with GM6001, a broad-spectrum MMP inhibitor, or NNGH (N-isobutyl-N-(4-methoxyphenylsulfonyl)-glycylhydroxamic acid), an MMP-3 inhibitor. The activated form of MMP-3 enhanced glycerol release as well as TNF-alpha protein secretion from 3T3-L1 adipocytes. The incubation of adipocytes with MMP-3 inhibited insulin-induced glucose uptake in adipocytes. Furthermore, a high-fat intake increased the expression of MMP-3, decreased the insulin-induced glucose uptake of adipocytes and induced expression of F4/80 in mesenteric fat tissue of C57BL/6 mice.Mphi may cause a pathological link with surrounding adipocytes through the secretion of MMP-3 followed by TNF-alpha expression in adipocytes in visceral fat tissue.

Esophageal rupture is a potentially mortal condition. Rapid and correct diagnosis, and urgent surgical treatment with esophagectomy is indicated, but conservative and other surgical treatments have also been reported recently. The treatment strategies for esophageal rupture are discussed here, based on our experiences with four cases during the last 10 years. They were admitted urgently and each was treated by a different method. Three of them underwent emergency operations, one undergoing primary closure of the ruptured esophagus, another received a T-tube insertion from the ruptured site with omental flap, and the third an esophagogastrectomy. The fourth case was treated conservatively. All patients survived and were discharged 36-144 days post treatment. One of them was readmitted for debridement of necrotic rib. In conclusion, the prompt and accurate diagnosis of esophageal rupture is crucial for a subsequent successful treatment. Conservative treatment or operation including esophagectomy will be determined by the severity of the condition.

Dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) are key enzymes for predicting the efficacy of 5-FU in the treatment of malignant tumors. However, 5-FU is not commonly commonly chosen for chemotherapeutic treatment of hepatocellular carcinoma (HCC) in practice. The aim of this study was to determine the activities of both DPD and TS in HCCs and corresponding liver parenchyma and to assess the correlation between the activities of these enzymes and clinicopathological features. The possibility of using 5-FU as a first-choice chemotherapeutic agent for HCC was also evaluated. The study material comprised 33 pairs of hepatocellular carcinoma and noncancerous liver samples. The DPD and TS activities were quantified by a radiometric enzymatic assay and a 5-fluoro-2′-deoxyuridine-5′- monophosphate (FdUMP) ligand-binding assay, respectively. Pathologically invasive HCCs tended to show higher DPD activity and lower TS activity with some exceptions. DPD activity was lower in the HCCs regardless of their clinical features than in the noncancerous liver parenchyma, whereas TS activity was generally lower in HCCs except for those with certain clinical features. HCCs with multiple nodules showed lower DPD activity and those with a diameter of more than 5 cm showed lower TS activity. In the noncancerous liver parenchyma, a gradual decrease in DPD activity and an increase in TS activity were associated with the age of the patient, liver damage and z-factor. Of 30 HCC samples, 10 exhibited comparatively low DPD and TS activity, and these could be considered 5-FU-sensitive HCC. DPD and TS activity may be affected by the clinicopathological status in both the HCC and the corresponding liver parenchyma. However, further investigation is necessary. Some HCC patients may be good candidates for 5-FU-based chemotherapy based on measurements of tumor levels of DPD and TS.

Giant cell tumor (GCT) of bone is a primary osteolytic tumor that is characterized by the formation of osteoclast-like giant cells. In addition to GCT of bone, extraskeletal GCT are known to be formed in several soft tissues. Giant cells in GCT of bone were suggested to be identical to osteoclasts, but the characterization of giant cells in extraskeletal GCT remains incomplete. In this study, a case of sarcomatoid hepatocellular carcinoma with osteoclast-like giant cells was analyzed. Immunohistochemistry revealed the expression of almost all markers of osteoclasts: tartrate-resistant acid phosphatase, CD68, CD51, CD54 and matrix metalloprotease-9, in osteoclast-like giant cells in the tumor. In situ hybridization revealed the expression of receptor activator of nuclear factor-kappa B (RANK) in the giant cells and receptor activator of nuclear factor-kappa B ligand (RANKL) in the tumor cells. The hepatic origin of the sarcomatoid hepatocellular carcinoma cells was confirmed by the expression of albumin. This is the first report suggesting that hepatocyte-derived cells possess the potential for osteoclastogenesis. In addition, these findings suggest that osteoclast-like cells in the hepatocellular carcinoma were formed by the same mechanism as osteoclastogenesis in bone.

Recipients for liver transplantation often have portosystemic shunts due to portal hypertension. It is an important problem whether such shunts should be ligated during operations. Ligating the shunts seems of benefit for increasing portal blood flow to the liver, but it is sometimes difficult technically, and it is invasive to the patient. We experienced a recipient with huge portosystemic shunts and no esophageal varices before living-related liver transplantation. Some shunts were ligated during operation to increase portal blood flow to the graft. Unfortunately, the patient suffered severe bleeding from esophagogastric varices after he underwent retransplantation owing to accidental liver failure. Based on our experience, extreme care should be exercised to avoid varicose bleeding after ligating the portosystemic shunts of liver transplantation patients.

Ischemia and reperfusion (IR) injury of the liver consists of two distinct phases. The first phase is caused by acute cellular injury at 3 to 6 h postreperfusion, which may be mainly induced by the increased production of oxygen radical species. The secondary, subacute, phase results from inflammatory responses at 18 to 24 h, leading to the progression of liver damage. The inflammatory response observed here is caused by proinflammatory cytokines and accumulating neutrophils which secrete oxidants, proteases, and so on. The production of proinflammatory cytokines, including chemokines and adhesion molecules, is regulated by transcriptional factors nuclear factor kappa B (NFKappaB), and AP-1. The progression of the injury is generated by the recruiting leucocytes which release oxidants and proteases. Recruitment and adhesion of neutrophils to the liver are accomplished by multiple steps in which many chemoattractants and adhesion molecules participate. Recent investigations suggest that calcium-dependent proteases, among various kinds of proteases, also play important roles in the aggravation of IR injury. Based on the mechanisms stated above, numerous strategies have been proposed as for prophylaxis and treatment. Most of these therapeutic strategies are derived from the inhibition of the production of oxygen radicals, inflammatory cytokines, and adhesion molecules; inhibition of leucocyte infiltration and elastase production; and inhibition of microcirculatory impairment, apoptosis-related molecules, and the breakdown of membrane phospholipids; and so on. Moreover, recent studies clarified that short periods of ischemia and subsequent reperfusion, termed ischemic preconditioning, exert a preventive effect against IR injuries in various organs, including the liver. Based on clarification of the candidates responsible for this phenomenon, pharmacological ischemic preconditioning has been proposed. In this review article, the authors outline the current progress in the understanding of and therapeutic strategies for hepatic IR injury.

The nationwide surveillance of antibacterial susceptibility to meropenem (MEPM) and other parenteral antibiotics against clinical isolates during 2012 in Japan was conducted. A total of 2985 strains including 955 strains of Gram-positive bacteria, 1782 strains of Gram-negative bacteria, and 248 strains of anaerobic bacteria obtained from 31 medical institutions were examined. The results were as follows; 1. MEPM was more active than the other carbapenem antibiotics tested against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae. MEPM was also active against most of the species tested in Gram-positive and anaerobic bacteria, except for multi-drug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA). 2. Of all species tested, there were no species, which MIC90 of MEPM was more than 4-fold higher than those in our previous studies in 2009 or 2006. Therefore, the tendency to increase in antimicrobial resistance rates was not observed. 3. MEPM resistance against Pseudomonas aeruginosa was 17.8% (56/315 strains). Compared to our previous results, it was the lowest than that in 2006 and 2009. 4. Carbapenem-resistant Klebsiella pneumoniae, and multi-drug-resistant Acinetobacter species, which emerged in worldwide, were not observed. 5. The proportion of extended-spectrum beta-lactamase (ESBL) strains was 6.2% (59/951 strains) in enterobacteriaceae, which increased compared with that of our previous studies in 2009 or before. Whereas, the proportion of metallo-beta-lactamase strains was 1.6% (5/315 strains) in P. aeruginosa, which was stable. In conclusion, the results from this surveillance suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem for serious infections treatment at present, 17 years passed after available for commercial use in Japan.

The patient was an 83-year-old woman who visited our hospital with a chief complaint of weight loss. Duodenal invasion by transverse colon cancer was suspected on abdominal computed tomography( CT). An extramural tumor invading the second portion of the duodenum was noted on upper gastrointestinal endoscopy, and a circumferential type 2 lesion was observed in the transverse colon on lower intestinal endoscopy. The pathological diagnosis of both lesions was well-differentiated adenocarcinoma, and right hemicolectomy and partial duodenectomy were performed. The invaded duodenal region was located on the oral side of the papilla of Vater, and the resection involved approximately one-third of the circumference of the second portion of the duodenum and was approximately 5 cm in length. The jejunum was elevated using the Roux- en-Y method, and side-to-side duodenojejunostomy was performed for reconstruction. On histopathological examination, the lesion was diagnosed as SI, N0, Stage II. Tumor markers with elevated levels before surgery showed a rapid decrease to within normal ranges. No evidence of recurrence or metastasis has been observed for 3 years after surgery. Although pancreatoduodenectomy is the standard surgical procedure for duodenal tumorous lesions, this approach is very invasive for elderly patients. Partial duodenectomy may be a favorable treatment option for cancers invading other organs, as was the case for this patient.

A series of piperazine ureas were designed, synthesized, and evaluated for their potential as novel orally efficacious fatty acid amide hydrolase (FAAH) inhibitors for the treatment of neuropathic and inflammatory pain. We carried out an optimization study of compound 5 to improve its in vitro FAAH inhibitory activity, and identified the 2-pyrimidinylpiperazine derivative 21d with potent inhibitory activity, favorable DMPK profile and brain permeability. Compound 21d showed robust and dose-dependent analgesic efficacy in animal models of both neuropathic and inflammatory pain.

Hormone replacement therapy (HRT) containing estrogens is generally used to relieve climacteric symptoms and to prevent osteoporosis and coronary heart disease [1], however, there has been increasing evidence of the HRT as the risk of hormone-dependent neoplasms including breast cancer [2], uterine endometrial cancer [3], ovarian cancer [4], and even lung cancer [5]. Noteworthy is mucinous cyst neoplasms (MCNs) of the pancreas, characterized by mucin-producing columnar epithelium supported by "ovarian-like" mesenchymal stroma, occur mostly in females expressing estrogen receptors [6, 7]. Although several reports regarding the closed relationship between MCNs and pregnancy [8, 9] might imply potential sex hormone-dependency of the MCNs [10], no correlation has been reported. This is the first case report of malignant MCN developing during continuous HRT after hysterectomy.

SUMMARY 1. AE0047 is a new dihydropyridine calcium antagonist with protective effects against cerebral ischaemia and the occurrence of stroke in several animal models. 2. In the present study we investigated whether AE0047 would improve the reduced cerebral blood flow (CBF) and oedema formation in cats subjected to middle cerebral artery (MCA) occlusion and compared it for efficacy with other dihydropyridine calcium antagonists with different moieties, such as nilvadipine an. nicardipine. 3. Middle cerebral artery occlusion reduced local CBF (1CBF), as measured by the hydrogen clearance method, while dry weight measurement showed that water content in the cortical tissues surrounding each 1CBF measurement electrode had increased after 4 h ischaemia. 4. Both AE0047 (10 μg/kg) and nilvadipine (30 μg/kg), given intravenously 20min after MCA occlusion, produced an approximate 10. hypotensive response and significantly increased 1CBF in severely and moderately ischaemic regions, grouped according to the initial reduced flow values. However, nicardipine (5 μg/kg bolus followed by infusion of 3 μg/kg per min for 60 min) failed to mitigate the reduction in 1CBF despite an increase in the ICBF of the contralateral cortex. In addition, AE0047 tended to prevent an increase in cortical water content in severely ischaemic regions, whereas water content in both nilvadipine- and nicardipine-treated groups tended to increase. 5. These results suggest that dihydropyridine calcium antagonists act differently on cerebral ischaemia and oedema formation in a manner dependent on their side-chain structures and that AE0047 effectively attenuates ischaemic brain damage without aggravatin. oedema.

RT-PCR and an in situ hybridization analyses demonstrated expression of prostaglandin EP2 receptor mRNA in the subfield of the dorsal horn of the rat spinal cord. The intensive signals from the cells labeled with digoxigenin-11-dUTP were scattered through this region and they corresponded to neurons. EP2 receptor-mediated signal transduction is expected to play roles in modulating some neuronal functions such as transmission of nociceptive reaction or motor reflex.

Summary Neuronal degeneration, distinguished morphologically and biochemically from necrosis, was induced in the dorsal horn of the lumbar spinal cord by chronic constriction of the sciatic nerve in rats. Neuronal apoptosis in the territory of the spinal cord which receives afferent excitatory inputs from the sciatic nerve was confirmed by TUNEL-staining and electrophoresis of genomic DNA. The morphological changes including the appearance of dark neurones, as identified by toluidine-blue staining, were almost completely blocked by 10 μg/kg of the prostaglandin E (EP) receptor agonist lipo-PGE 1 , incorporating PGE 1 in lipid microspheres for chemical stability and targeted delivery, but not by 10 μg/kg of carbacyclin a prostacyclin (IP) receptor agonist. Lipo-PGE] also blocked the “ladder type” fragmentation of genomic DNA extracted from tissue in the affected area of the spinal cord. Since the regional blood flow in the subfield of the spinal cord was neither influenced by the chronic injury nor by application of the vasodilative prostaglandin, the effect of lipo-PGE 1 must have been achieved via another mechanism. These results demonstrate that the neuronal apoptosis in spinal cord induced by sciatic constriction injury is more effectively inhibited by the EP receptor agonist PGE 1 than by the IP receptor agonist carbacyclin.

The carbon-hydride energy storage system stores hydrogen, which is difficult to handle, in the form of the stable liquid methylcyclohexane, thereby making it easy to transport and keep hydrogen in long-term storage as an energy medium. Since this makes it easy to convert energy into hydrogen for storage and reuse, it can contribute to the stable supply of renewable energy such as wind power, which fluctuates greatly. Expectations are high for this system as a means of improving Japan's energy self-sufficiency ratio by helping expand the use of renewable energy and accelerating the achievement of a low-carbon hydrogen society. Based on a 2011 contract with the National Institute of Polar Research, we have been working in the city of Nikaho in Akita Prefecture on the development of a practical version of this system while acquiring basic data that will, among other things, help improve the energy self-sufficiency ratio of Antarctica in the future.