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Dysbiosis in the intestinal microflora can affect the gut production of microbial metabolites, and toxic substances can disrupt the barrier function of the intestinal wall, leading to the development of various diseases. Decreased levels of Clostridium subcluster XIVa (XIVa) are associated with the intestinal dysbiosis found in inflammatory bowel disease (IBD) and

Western-style high-fat/high-sucrose diet (HFHSD) changes gut microbiota and bile acid (BA) profiles. Because gut microbiota and BAs could influence each other, the mechanism of changes in both by HFHSD is complicated and remains unclear. We first aimed to clarify the roles of BAs in the HFHSD-induced change of gut microbiota. Then, we studied the effects of the changed gut microbiota on BA composition and liver function. Male wild-type (WT) and human-like Cyp2a12/Cyp2c70 double knockout (DKO) mice derived from C57BL/6J were fed with normal chow or HFHSD for 4 weeks. Gut microbiomes were analyzed by fecal 16S ribosomal RNA gene sequencing, and BA composition was determined by liquid chromatography-tandem mass spectrometry. The DKO mice exhibited significantly reduced fecal BA concentration, lacked muricholic acids, and increased proportions of chenodeoxycholic and lithocholic acids. Despite the marked difference in the fecal BA composition, the profiles of gut microbiota in the two mouse models were quite similar. An HFHSD resulted in a significant increase in the BA pool and fecal BA excretion in WT mice but not in DKO mice. However, microbial composition in the two mouse models was drastically but similarly changed by the HFHSD. In addition, the HFHSD-induced change of gut microbiota inhibited BA deconjugation and 7α-dehydroxylation in both types of mice, which improved chronic liver injury observed in DKO mice. Conclusion: The HFHSD itself causes the change of gut microbiota due to HFHSD, and the altered composition or concentration of BAs by HFHSD is not the primary factor. On the contrary, the gut microbiota formed by HFHSD affects BA composition and ameliorates liver injury in the mouse model with human-like hydrophobic BA composition.

Riluzole, a drug used in the management of amyotrophic lateral sclerosis (ALS), is associated with a high incidence of liver failure. It is imperative to determine risk factors and severity of liver injury in patients taking riluzole to devise an appropriate treatment regimen. We, therefore, studied risk factors for liver injury in ALS patients who were prescribed riluzole at Kitasato University East Hospital from 1999 to 2015. Of the 222 patients enrolled in this study, 113 and 109 patients were diagnosed with mild to moderate (grade 1 or 2) and without (grade 0) liver injury, respectively. Prediction of risk factors was determined using binary logistical regression analyses. The results showed that 50.9% (n=113) of ALS patients developed mild to moderate liver injury; 71.7% and 53.1% of patients were concurrently using CYP1A2 inhibitors (p=0.005) and diclofenac (p=0.032), respectively; 55.8% of patients with liver injury had a history of smoking (p=0.011). Multivariate analyses revealed that the concurrent use of CYP1A2 inhibitors [odds ratio (OR) 2.152, 95% confidence interval (CI) 1.225-3.780, p=0.008] and history of smoking (OR 1.938, 95% CI 1.125-3.340, p=0.017) were independent risk factors for liver injury in patients receiving riluzole. In conclusion, treatment of ALS patients with riluzole, smoking habits, and concurrent use of CYP1A2 inhibitors are independent liver injury risk factors. Further studies on liver injury are warranted in ALS patients treated with riluzole to comprehensively understand the underlying mechanisms of riluzole-associated liver toxicity.

Non-steroidal anti-inflammatory drugs (NSAIDs) or low-dose aspirin (LDA) are the most common causes of drug-induced gastroduodenal ulcer and We investigated preventive treatment with use of concomitant anti-ulcer drugs and the clinical features of gastroduodenal ulcer in cases treated with these drugs. Patients with gastroduodenal ulcer and patients with bleeding were classified into 3 groups: LDA, non-aspirin NSAIDs, and those taking neither aspirin nor NSAIDs. Chronological changes over the past 16 years (1st–5th period) were investigated. The status of prevention of ulcer and clinical features were examined. From January 2002 to December 2018, the ratio of all patients taking NSAIDs and LDA increased significantly until 3rd period (p

The bile acid (BA) composition in mice is substantially different from that in humans. Chenodeoxycholic acid (CDCA) is an end product in the human liver; however, mouse Cyp2c70 metabolizes CDCA to hydrophilic muricholic acids (MCAs). Moreover, in humans, the gut microbiota converts the primary BAs, cholic acid and CDCA, into deoxycholic acid (DCA) and lithocholic acid (LCA), respectively. In contrast, the mouse Cyp2a12 reverts this action and converts these secondary BAs to primary BAs. Here, we generated Cyp2a12 KO, Cyp2c70 KO, and Cyp2a12/Cyp2c70 double KO (DKO) mice using the CRISPR-Cas9 system to study the regulation of BA metabolism under hydrophobic BA composition. Cyp2a12 KO mice showed the accumulation of DCAs, whereas Cyp2c70 KO mice lacked MCAs and exhibited markedly increased hepatobiliary proportions of CDCA. In DKO mice, not only DCAs or CDCAs but also DCAs, CDCAs, and LCAs were all elevated. In Cyp2c70 KO and DKO mice, chronic liver inflammation was observed depending on the hepatic unconjugated CDCA concentrations. The BA pool was markedly reduced in Cyp2c70 KO and DKO mice, but the FXR was not activated. It was suggested that the cytokine/c-Jun N-terminal kinase signaling pathway and the pregnane X receptor-mediated pathway are the predominant mechanisms, preferred over the FXR/small heterodimer partner and FXR/fibroblast growth factor 15 pathways, for controlling BA synthesis under hydrophobic BA composition. From our results, we hypothesize that these KO mice can be novel and useful models for investigating the roles of hydrophobic BAs in various human diseases.

Objective Since the majority of direct-acting antivirals (DAAs) that are used in the treatment of hepatitis C virus (HCV) infection are mainly metabolized by CYP3A4, it is hypothesized that inter-individual differences in CYP3A4 activity may be associated with the bioavailability of these agents. Methods The level of serum 4β-hydroxycholesterol (4βHC), a surrogate marker of CYP3A4 activity, was determined by LC-MS/MS in samples obtained from patients with HCV infection (CHCs) as well as healthy control subjects (CTLs). Serum samples obtained from patients treated with either asunaprevir/daclatasvir (ASV/DCV) or ombitasvir/paritaprevir/ritonavir (OTV/PTV/r) were used for additional assays. Results The serum 4βHC level in CHCs was significantly higher than that in CTLs, and a gender difference was seen among CHCs. In patients treated with OTV/PTV/r, the serum 4βHC level was observed to gradually decrease during the treatment period. In the cohort treated with ASV/DCV, 4 of 83 patients showed virological treatment failure. In pretreatment testing, an Invader assay detected a low prevalence of resistance-associated variants in these four patients. The average serum concentration of DCV/ASV in the treatment-failed group tended to be lower than that in the sustained virological response (SVR) group. The pretreatment serum 4βHC level in patients with treatment failure was significantly higher than that in patients with an SVR but in whom the prevalence of resistance-associated variants was low in the pretreatment setting. Conclusion The evaluation of CYP3A4 activity by measuring 4βHC before treatment may provide additional information that can potentially be used to select cost- and efficacy-optimized treatment of HCV.

A 66-year-old Japanese male with a history of Behcet disease exhibited oral and genital ulcers, and a round deep ileocecal ulcer. He was treated with a combination of mesalazine and 20 mg/day of prednisolone (PSL), but was only partially responsive to PSL and we were not able to reduce the steroid dosage. Adalimumab was also administered. However, the ulcer was not completely responsive, and weaning the patient off PSL remained impossible. In contrast, additional treatment with clarithromycin completely healed the refractory active ulcer and left only a scar. Furthermore, the ulcer has since maintained the scar stage despite successfully weaning the patient from PSL.

Background and Aim: Non-steroidal anti-inflammatory drugs (NSAIDs) or low-dose aspirin (LDA) are the most common causes of drug-induced gastroduodenal ulcer. We investigated preventive treatment with use of concomitant anti-ulcer drugs and the clinical features of gastroduodenal ulcer in cases treated with these drugs.Method: A total of 2,332 patients with gastroduodenal ulcer and 241 patients with bleeding gastroduodenal ulcer were classified into 3 groups: those taking LDA, those taking non-aspirin NSAIDs (hereinafter referred to as NSAIDs), and those taking neither aspirin nor NSAIDs. Chronological changes in the percentage of each group and the change over the past 15 years were investigated. The status of prevention of ulcer and clinical features were examined in 264 patients with gastroduodenal ulcer taking NSAIDs or LDA, including 107 bleeding cases, in the past 8 years.Results: From January 2002 to December 2017, the percentage of all patients taking LDA increased until 2013, but from 2014, the percentage of patients taking LDA decreased in those with a bleeding ulcer. The percentage of patients taking NSAIDs decreased from 2002 in those with a bleeding ulcer. Among the 264 patients with gastroduodenal ulcer and the 107 patients with a bleeding ulcer taking NSAIDs and LDA, 16 (6%) and 9 (8%), respectively, were receiving preventive treatment with concomitant anti-ulcer drugs. The percentages of patients taking LDA and other antiplatelet drugs in patients with bleeding gastroduodenal ulcer were significantly higher than those in patients with non-bleeding. Conclusion: Although the percentages of patients with gastroduodenal ulcer taking NSAIDs or LDA have not recently increased in real-world practice, preventive treatment in these patients is still low. This low rate of prevention in cases of non-bleeding and bleeding gastroduodenal ulcer suggests the need to enlighten physicians about preventive treatment because drug withdrawal of LDA has a high risk of cardiovascular and cerebrovascular events

Background Dysbiosis, especially a reduced Clostridium subcluster XIVa (XIVa), has been reported in several gastrointestinal diseases. Since XIVa is thought to be the main bacterial cluster that metabolizes bile acids (BAs) in the human intestine, we hypothesized that the BA profile in feces, and possibly in serum, could be a convenient biomarker for intestinal XIVa activity. Methods First, blood and feces were collected from 26 healthy controls and 20 patients with gastrointestinal diseases, and the relationships among fecal microbiomes and fecal and serum BA compositions were studied. Second, serum BA compositions of 30 healthy controls and the remission and exacerbation states of 14 Crohn's disease (CD) and 12 ulcerative colitis (UC) patients were compared. Fecal microbiomes were analyzed by terminal restriction fragment length polymorphism analysis, and BA compositions were quantified by HPLC-MS/MS. Results The highest positive correlation was observed between the fecal XIVa proportion and fecal unconjugated deoxycholic acid (DCA)/(DCA+unconjugated cholic acid [CA]) (r = 0.77, P < 0.0001) or serum DCA/(DCA+CA) (r = 0.52, P < 0.001). Diurnal variation in serum XIVa candidate markers also showed that DCA/(DCA+CA) was most stable and not affected by the contraction of the gallbladder. Serum DCA/(DCA+CA) was not significantly different between remission and exacerbation states in either CD or UC patients, but was significantly reduced in those in the remission state of CD and the remission and exacerbation states of UC compared with healthy controls (P < 0.05). Conclusions Decreased XIVa exhibits a strong correlation with reduced intestinal BA metabolism. Fecal and serum DCA/(DCA+CA) could be useful surrogate markers for the intestinal proportion of XIVa.

Bezafibrate is a widely used hypolipidemic agent and is known as a ligand of the peroxisome proliferator-activated receptors (PPARs). Recently this agent has come to be recognized as a potential anticholestatic medicine for the treatment of primary biliary cirrhosis (PBC) that does not respond sufficiently to ursodeoxycholic acid (UDCA) monotherapy. The aim of this study was to explore the anticholestatic mechanisms of bezafibrate by analyzing serum lipid biomarkers in PBC patients and by cell-based enzymatic and gene expression assays. Nineteen patients with early-stage PBC and an incomplete biochemical response to UDCA (600 mg/day) monotherapy were treated with the same dose of UDCA plus bezafibrate (400 mg/day) for 3 months. In addition to the significant improvement of serum biliary enzymes, immunoglobulin M (IgM), cholesterol, and triglyceride concentrations in patients treated with bezafibrate, reduction of 7α-hydroxy-4-cholesten-3-one (C4), a marker of bile acid synthesis, and increase of 4β-hydroxycholesterol, a marker of CYP3A4/5 activity, were observed. In vitro experiments using human hepatoma cell lines demonstrated that bezafibrate controlled the target genes of PPARα, as well as those of the pregnane X receptor (PXR); down-regulating CYP7A1, CYP27A1, and sinusoidal Na+/taurocholate cotransporting polypeptide (NTCP), and up-regulating CYP3A4, canalicular multidrug resistance protein 3 (MDR3), MDR1, and multidrug resistance-associated protein 2 (MRP2). Conclusion: Bezafibrate is a dual PPARs/PXR agonist with potent anticholestatic efficacy in early-stage PBC patients with an incomplete biochemical response to UDCA monotherapy. (HEPATOLOGY 2013)

To date, many studies have been conducted using 25-hydroxycholesterol, which is a potent regulator of lipid metabolism. However, the origins of this oxysterol have not been entirely elucidated. Cholesterol 25-hydroxylase is one of the enzymes responsible for the metabolism of 25-hydroxycholesterol, but the expression of this enzyme is very low in humans. This oxysterol is also synthesized by sterol 27-hydroxylase (CYP27A1) and cholesterol 24-hydroxylase(CYP46A1), but it is only a minor product of these enzymes. We now report that CYP3A synthesizes a significant amount of 25-hydroxycholesterol and may participate in the regulation of lipid metabolism. Induction of CYP3A by pregnenolone-16α-carbonitrile caused the accumulation of 25-hydroxycholesterol in a cell line derived from mouse liver. Furthermore, treatment of the cells with troleandomycin, a specific inhibitor of CYP3A, significantly reduced cellular 25-hydroxycholesterol concentrations. In cells that overexpressed human recombinant CYP3A4, the activity of cholesterol 25-hydroxylation was found to be higher than that of cholesterol 4β-hydroxylation, a known marker activity of CYP3A4. In addition, 25-hydroxycholesterol concentrations in normal human sera correlated positively with the levels of 4β-hydroxycholesterol (r = 0.650, P < 0.0001, n = 78), but did not significantly correlate with the levels of 27-hydroxycholesterol or 24S-hydroxycholesterol. These results demonstrate the significance of CYP3A on the production of 25-hydroxycholesterol.

AIM Infection with hepatitis C virus (HCV) is the leading cause of liver cirrhosis that develops into hepatocellular carcinoma. Previous studies have shown in vitro that lipids within hepatocytes are crucially important for a series of HCV infection-proliferation-release processes. On the other hand, in the patients with HCV, the serum total cholesterol (Total-C) and low-density lipoprotein cholesterol (LDL-C) levels have been reported to be lower. We conducted an epidemiological survey of a large cohort and investigated whether the lower serum lipid levels were caused by a direct or the secondary effects of HCV infection (i.e. hepatic damage or nutritional disorder). METHODS Among 146 857 participants (male, 34%; female, 66%) undergoing public health examinations between 2002 and 2007 in Ibaraki Prefecture, Japan, the HCV positive rates determined by HCV antibody/antigen and/or RNA tests were 1.37% and 0.67% in males and females, respectively. RESULTS In addition to Total-C and LDL-C, serum high-density lipoprotein cholesterol and triglyceride concentrations were also significantly lower in the HCV positive subjects compared with the negative subjects, regardless of sex, age or nutritional state evaluated by body mass index. Multivariate analysis showed that HCV infection was the strongest among the factors to be significantly associated with the lower level of these lipids. Particularly, the hypolipidemia was also confirmed in the HCV positive subjects with normal aminotransferase levels (alanine aminotransferase ≤30 and aspartate aminotransferase ≤30). CONCLUSION This epidemiological survey in a large Japanese cohort suggests that the HCV infection itself might directly cause hypolipidemia, irrespective of host factors including age, hepatic damage and nutritional state.

Opioid rotation has been proposed for management of cancer pain. No studies directly investigating dose equivalence between morphine injection (continuous IV administration) and the transdermal fentanyl patch have been reported. Therefore, we examined dose conversion ratios in patients undergoing opioid rotation from morphine injection to fentanyl patches. The subjects consisted of 45 patients admitted to Kitasato University East Hospital. Medical records were consulted to determine the "basic dose of morphine injection immediately prior to rotation" and the "basic dose of fentanyl patch after rotation". Equivalent doses and conversion ratios obtained with the expression of (daily dose of morphine injection (mg)/daily delivered dose of fentanyl patch (mg)) were determined from the relationship between the data by regression analysis. The regression equation obtained was Y=50.882X-13.96, r²=0.8922, where X and Y are daily doses of morphine injection and fentanyl patch, respectively. Equivalent doses and conversion ratios for daily dose of morphine injection (mg): daily delivered dose of fentanyl patch (mg) (patch dose mg/3 days) were 16.6 mg: 0.6 mg (2.5 mg)=28:1, 47.1 mg: 1.2 mg (5 mg) = 39:1 and 169.2 mg: 3.6 mg (15 mg)=47:1. In other reports, the ratio of morphine vs. fentanyl at 50:1 had no relation to the dose. While the present study suggested that in opioid rotation from low dose, 50:1 is not enough for the fentanyl patch. The dose conversion ratio of morphine injection to fentanyl patch was different at the low doses and high doses of morphine.

We describe a new sensitive and specific method for the quantification of serum malonate (malonic acid, MA), which could be a new biomarker for de novo lipogenesis (fatty acid synthesis). This method is based upon a stable isotope-dilution technique using LC-MS/MS. MA from 50 microl of serum was derivatized into di-(1-methyl-3-piperidinyl)malonate (DMP-MA) and quantified by LC-MS/MS using the positive electrospray ionization mode. The detection limit of the DMP-MA was approximately 4.8 fmol (500 fg) (signal-to-noise ratio = 10), which was more than 100 times more sensitive compared with that of MA by LC-MS/MS using the negative electrospray ionization mode. The relative standard deviations between sample preparations and measurements made using the present method were 4.4% and 3.2%, respectively, by one-way ANOVA. Recovery experiments were performed using 50 microl aliquots of normal human serum spiked with 9.6 pmol (1 ng) to 28.8 pmol (3 ng) of MA and were validated by orthogonal regression analysis. The results showed that the estimated amount within a 95% confidence limit was 14.1 +/- 1.1 pmol, which was in complete agreement with the observed X(0) = 15.0 +/- 0.6 pmol, with a mean recovery of 96.0%. This method provides reliable and reproducible results for the quantification of MA in human serum.

Aim: Although the anti-hepatitis C virus (HCV) antibody test has been recommended to the whole Japanese population, most countries have not implemented it. The present study aims to re-evaluate the usefulness of markers examined in the general health examination for the initial screening of HCV carriers. Methods: Of the overall population, 25 142 individuals (8876 males, 16 266 females) participated in health examinations with HCV tests in 2005, and the most commonly associated markers for HCV-positive subjects were explored by multivariate analysis, based on blood biochemical, physical, sphygmomanometric and hematological parameters. Thereafter, the efficiencies of the markers were estimated from a total population of 85 013 individuals (29 502 males, 55 511 females) in 2003–2005. Results: The most significantly associated markers for HCV positivity were aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Optimal limits of ALT and AST by receiver–operator characteristic (ROC) analysis were 24 and 27 IU (male, 33 and 28 IU; female, 22 and 26 IU), respectively. However, one-quarter of HCV carriers were not found to be positive using the optimal limits of aminotransferases. Conclusion: The present study confirmed the limitation of serum aminotransferase levels as markers of HCV for primary screening. Therefore, at present, an anti-HCV antibody test is required for the efficient screening of HCV carriers in all health examinations.

This study was designed to investigate: 1) the distribution of Neobenedenia girellae (Capsalidae) on host amberjack Seriola dumerili (Carangidae) and growth of the parasite during infection; 2) the influence of different N. girellae exposure levels on S. dumerili growth; and 3) the histopathological skin changes of S. dumerili infected with N. girellae at the different parasite densities. N. girellae were mostly found on pelvic and pectoral fins of S. dumerili at 1 and 3 days after exposure of S. dumerili to N. girellae oncomiracidia. The parasites were primarily found on the skin surface at 6 days after exposure, and all mature N. girellae specimens were found on the skin surface at 9 and 12 days after exposure. These results show that N. girellae migrated from the fins to the skin surface during parasite growth. The growth of S. dumerili infected with N. girellae was significantly slower compared with that of uninfected fish when the number of infected parasites was more than 0.285 ± 0.042/cm2 fish surface showing that N. girellae infections hindered the growth of the host fish. In addition, the feed conversion ratio was positively correlated with the number of infected parasites on fish. Furthermore, the thickness of S. dumerili epidermis with attached N. girellae was negatively correlated with both the N. girellae infection duration and the number of infected parasites. Thus, larger N. girellae caused more severe infections on host S. dumerili. These results in this study could be applied to control N. girellae infections and to promote stable S. dumerili production in aquaculture.

The infinite dilution activity coefficients of C1 to C5 alcohols, acetone, 2-butanone, acetylacetone, toluene, and xylene isomers in 4-methyl-N-butylpyridinium tetrafluoroborate ([bmpy][BF4]) and 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]) were measured by gas−liquid chromatography (GC) from (306.6 to 334.8) K and atmospheric pressure. In these measurements with GC, [bmpy][BF4] or [bmim][PF6] was used as a stationary phase. The infinite dilution activity coefficients in this work were not affected by the gas flow rate. It was found that the values of 1-propanol, 1-butanol, and 1-pentanol for the infinite dilution activity coefficients in [bmim][PF6] were larger than those for their isomers. For xylene isomers, the activity coefficients of m-xylene are the largest. The effects of the cation and anion species composing ionic liquid on the activity coefficients were also investigated. The activity coefficients for alcohols, ketones, and aromatic hydrocarbons in the ionic liquids were affect...

A flow-type method was adopted to measure the vapor–liquid equilibria for methanol + methyl laurate and methanol + methyl myristate systems at 493–543 K, near the critical temperature of methanol (Tc = 512.64 K), and 2.16–8.49 MPa. The effect of temperature and fatty acid methyl esters to the phase behavior was discussed. The mole fractions of methanol in liquid phase are almost the same for both systems. In vapor phase, the mole fractions of methanol are very close to unity at all temperatures. The present vapor–liquid equilibrium data were correlated by PRASOG. A binary parameter was introduced to the combining rule of size parameter. The binary parameters of methanol + fatty acid methyl ester systems were determined by fitting the present experimental data. The correlated results are in good agreement with the experimental data. The vapor–liquid equilibria for methanol + methyl laurate + glycerol and methanol + methyl myristate + glycerol ternary systems were also predicted using the methanol + fatty acid methyl ester binary parameters. The mole fractions of methanol in vapor phase are around unity even if glycerol is included in the systems.

Yellowtail ascites virus (YTAV) is the causative agent of ascites and deformity in fish and causes serious losses to the fish-farming industry of yellowtail fry and fingerling Seriola quinqueradiata in Japan. In 2006, cultured yellowtail died from ascites in Kochi, Japan. We isolated and characterized a virus from the diseased fish. Based on the pathogenicity, culture characteristics, morphological features, RT-PCR results targeting VP2/NS region, phylogeny based on the VP1 amino acid sequence, and immunochemical reactivity of structural proteins, the virus isolate was identified as YTAV (designated as YTAV-06). YTAV-06 was a more virulent isolate than YTAV Y-6, isolated originally from yellowtail with ascites. To our knowledge, this is the first report describing that YTAV isolates may vary in their virulence.

We have developed a highly sensitive and specific method for quantification of salivary 3-hydroxybutyrate (3HB), 3-hydroxyisobutyrate (3HIB), 3-hydroxy-3-methylbutyrate (3HMB) and 2-hydroxybutyrate (2HB), which could be new non-invasive biomarkers for catabolic pathways of fatty acids/ketogenic amino acids, valine, leucine, and methionine/threonine/α-ketobutyrate, respectively. The four hydroxybutyrates (3HB, 3HIB, 3HMB, and 2HB) were extracted from 5 µl of saliva, converted to 2-pyridylmethyl (2PM) ester derivatives, and measured by liquid chromatography–tandem mass spectrometry in positive electrospray ionization mode. [13C4]3HB was used as an internal standard. The detection limits for the 2PM esters were

SummaryBackground The sebaceous gland and the follicular bulge region have important role in biology of the hair. They initiate destruction of the hair follicle both in human and animal models in certain conditions. The morphometric feature of the sebaceous gland is not well understood so as the distribution of the bulge stem cells in pathological conditions of male pattern hair loss or androgenic alopecia (AGA). Objectives The authors perform morphometric analysis of the sebaceous gland in AGA patients and also study distribution of the follicular stem cells in the bulge region in these populations. Methods Two hundred and fifty cases of glass slide specimen from Japanese patients with male pattern hair loss were reviewed. Among these, 23 cases of the longitudinal (vertical) sections of the scalp skin with diagnosis of AGA were found and analyzed for the morphometric characteristics. Each sebaceous gland area was measured using NIH imagej system and statistically analyzed. For the identification of the follicular bulge region, an immunohistochemistry using anticytokeratin 15 (C8/144B clone) was carried out in the cases of AGA. Results The sebaceous gland area of the AGA group was noticeably increased, while the size of each sebaceous gland remains unchanged. It has more lobules in the hair follicular unit in the AGA population. In the immunohistochemistry, the follicular stem cells are present in the bulge regions in cases of AGA. Conclusions The overgrowth (multilobulation) of the sebaceous gland and relative preservation of the follicular stem cells suggest that the changes in the sebaceous gland could be an important factor in the pathology of AGA.

Hypercholesterolemia is an important paraneoplastic syndrome in patients with hepatoma, but the nature of this defect has not yet been identified. We investigated the molecular mechanisms of hypercholesterolemia in a hepatoma-bearing rat model. Buffalo rats were implanted in both flanks with Morris hepatoma 7777 (McA-RH7777) cells. After 4 weeks, tumor weight was 5.5+/-1.7 g, and serum cholesterol level increased from 60+/-2 to 90+/-2 mg/dL. Protein and mRNA expression of the ATP-binding cassette transporters A1 and G1 (ABCA1 and ABCG1) was markedly higher in tumors than in livers. These increases were associated with activation of liver X receptor alpha (LXRalpha) as a result of the increased tissue oxysterol concentrations. The accumulation of oxysterols in the hepatomas appeared to be caused mainly by the upregulation of cholesterol biosynthesis, despite the increased tissue sterol concentrations. Overexpression of the sterol regulatory element-binding protein (SREBP) processing system relative to sterol concentration contributed to the resistance to sterols in this tumor. In addition, bile acid biosynthesis was inhibited despite the reduced expression of the small heterodimer partner (SHP) and activated LXRalpha, which also appeared to contribute to the accumulation of oxysterols followed by the acceleration of cholesterol efflux. In conclusion, hypercholesterolemia in McA-RH7777 hepatoma-bearing rats was caused by increased cholesterol efflux from tumors as a result of activation of LXRalpha. Overexpression of the SREBP processing system contributed to the activation of LXRalpha by maintaining high oxysterol levels in tissue.

Infectious pancreatic necrosis virus (IPNV), a member of the genus Aquabirnavirus and family Birnaviridae, is an unenveloped icosahedral virus with two segments of double-stranded RNA. IPNV causes acute infection in salmonid fry and fingerlings with high mortality. However, this mortality is low as the age increases and survivors become IPNV-carrier fish. In this study, IPNV persistent infection was established in rainbow trout with no clinical signs or mortality. TUNEL staining and immunohistochemistry showed that IPNV antigen-positive cells did not have an apoptotic nucleus in almost all tissue sections and leucocyte smears, indicating that apoptosis was not induced in IPNV antigen-positive cells. The IPNV genome detected by in situ RT-PCR was more frequent than detection of the IPNV antigen by immunohistochemistry in the kidney, spleen, and liver. This result implies that the successive replication would not occur in many IPNV-infected cells. Further, apoptotic cells were predominant in the tissue sections where the signal-positive cells were frequently detected. Therefore, the presence of apoptosis in this study might be associated with host defense mechanisms, which eliminates IPNV-infected cells by the recognition of IPNV genome at the early stage of infection.

Marine birnavirus (MABV) is a member of the genus Aquabirnavirus of the family Birnaviridae. MABV is an unenveloped icosahedral virus about 60 nm in diameter with two genomes of double-stranded RNA. MABV adsorbed not only onto the cell surfaces of susceptible (CHSE-214 and RSBK-2) cells but also onto resistant (FHM and EPC) cells. Furthermore, the virus entered into the cytoplasm through the endocytotic pathway in CHSE-214, RSBK-2 and FHM cells but did not penetrate EPC cells. The virus was found to bind to an around 250 kDa protein on CHSE-214, RSBK-2, FHM and EPC cells. The syntheses of viral proteins pVP2, NS and VP3 and further proteolytic processing after viral infection were examined by using Western blot analysis. pVP2, NS and VP3 were detected in the cytosolic fractions of CHSE-214, RSBK-2 and FHM cells at 4 h after infection. At this time, VP3 underwent further proteolytic processing in the cytosolic fractions of CHSE-214 and RSBK-2 cells. The expression of pVP2, NS and VP3 increased and pVP2 and NS also underwent further proteolytic processing similar to VP3 in the cytosolic fractions of CHSE-214, RSBK-2 and FHM cells at 8 h after infection. The further proteolytic processing of VP3 was detected in the nuclear fractions of CHSE-214, RSBK-2, but VP3 was detected as a single band in the nuclear fraction of FHM cells. pVP2 and NS were detected as thin bands only in the nuclear fractions of CHSE-214 cells. The results of Western blot analysis demonstrated that pVP2, NS and VP3 are localized in the nuclear fraction when they were independently expressed in CHSE-214, RSBK-2, FHM and EPC cells. The expression pattern in the cytosolic fraction was identical among the four cell lines when pVP2 and NS were independently expressed. However, pVP2 and NS were not detected in the nuclear fraction of CHSE-214 cells. Further proteolytic processing of VP3 was detected in both cytosolic and nuclear fractions of RSBK-2 ,FHM and EPC cells (Low level in EPC cell), but not in CHSE-214 cells when VP3 was independently expressed. Then, the processes of preVP2 to form morphological assemblages in the presence of VP3 or the cleavage of VP3 into two proteins in CHSE-214 cells were studied. When preVP2- and VP3 were co-expressed, virion like particles (64 nm, diameter) were observed close to the nuclear membrane by electron microscopy. The co-expression of preVP2 and the cleaved VP3 proteins led to an efficient assembly of tubules (22 nm, diameter). Further important finds will be obtained by this infection system using 4 fish cell lines in the next couple of years.

Postoperative nausea and vomiting (PONV) with morphine therapy develops in more than 60% of patients after surgery, markedly reducing patient QOL. The prophylactic effect of several antiemetics has already been studied, but evaluations, and even those using the same drug, are not uniform. The present research involved a meta-analysis of randomized controlled trials on prophylactic drug therapy for PONV in patients receiving morphine for the treatment of postoperative pain. The efficacy of the prophylactic administration of the drugs was examined. As a result, meta-analysis of five drugs was possible and the evidence of efficacy was shown for three drugs ranked in order of an increasing odds ratio (OR) and confidence interval (CI): dexamethasone (OR: 0.23, 95% CI: 0.15-0.35, p < 0.00001), droperidol (OR: 0.27, 95% CI: 0.21-0.34, p < 0.00001), and metoclopramide (OR: 0.48, 95% CI: 0.30-0.75, p < 0.001). These results suggest that the three drugs are effective in prophylactic treatment for PONV. Of them, dexamethasone used as a prophylactic drug for PONV provided the best results. Dexamethasone was shown to reduce the incidence of PONV from 66-80% to 16-50% with a dose of 1.25 to 10 mg and to be suitable as a first drug of choice.

The population pharmacokinetics on tazobactam/piperacillin (TAZ/PIPC; 1:8, 4.5 g x 3) was analyzed in Japanese patients with community-acquired pneumonia using the Nonlinear Mixed Effect Model version VI. Analysis by the one-compartment model yielded the following results for PIPC: total clearance (CL) = 8.22+(Ccr-71.4) x 0.0561 (L/hr), distribution volume (Vd) = 13.7 (L). The pharmacokinetic parameters for TAZ were: CL = 8.67 + (Ccr-71.4) x 0.0682 (L/hr), Vd = 14.4 (L). Of the pharmacokinetic parameters of PIPC, CL included Ccr as a variation factor, whereas the Vd included no variation factor. Because PIPC is excreted into the urine in the unchanged form, its pharmacokinetic factors seem to reflect the renal function status. In this study of patients with community-acquired pneumonia, the mean Vd per body weight was 0.26 L/kg, and the results suggested an increase of the Vd in patients with community-acquired pneumonia as compared with the value in healthy adults.

A bibliographic search was conducted of cases of Candida endophthalmitis reported in Japan and overseas between 2000 and 2011, in the Japana Centra Revuo Medicina Website of Japan Medical Abstracts Society (domestic reports) and MEDLINE (overseas reports). The investigation yielded 42 reports in domestic journals (49 cases ; hereinafter referred to as domestic cases) and 39 reports in journals published overseas (46 cases ; hereinafter referred to as overseas cases). The isolation rate of Candida albicans in the domestic cases was 65.3%, and that in the overseas cases was 71.7%. The initial treatment for the Candida endophthalmitis was fluconazole (FLCZ) therapy in 51.0% of the domestic cases and 38.1% of the overseas cases. Domestic reports suggested the effectiveness of FLCZ therapy for stage II cases, and of vitrectomy for stage III and IV cases. Reports from overseas, on the other hand, suggested the effectiveness of amphotericin B (AMPH-B) or voriconazole (VRCZ) therapy for stage II cases, and of vitrectomy for stage III and IV cases.

A series of numerical experiments on the failure process of homogeneous rock slope has been performed using Boundary Element Method and Displacement Discontinuity Method, and then the effect of initial rock stress on the stability of homogeneous rock slope has been discussed.Firstly, the distribution of tangential stress along the surface of rock slope has been analysed and the condition of fracture initiation has been formulated. The stability factor at the fracture initiation.of rock slope is affected by not only the slope angle and the rock strength but also the initial rock stress. Increasing the ratio of horizontal to vertical stresses at the pre-excavation, the value of the stability factor decreases and the location of fracture changes from the toe of slope to the upper surface.Secondly, failure process of rock slope has been analyzed on the basis of the Fracture Mechanics. It is made clear that the failure process and the failure mode are also affected significantly by the initial stress state as well as the rock strength, the frictional characteristics of the crack surface and the slope angle. Furthermore, it is pointed out that the critical value of the stability factor is rather independent of the initial stress state.

Transnasal endoscopy is generally recognized as a less painful endoscopic procedure. The pretreatment procedure, particularly anesthesia of the nasal cavities, is reported to be vitally important for alleviating pain.The study comprised 120 patients treated by the spray method, 203 treated by the stick method, and 100 treated by the stick + pharyngeal anesthesia method between April 2005 and March 2009. We investigated the pain levels using three pretreatment methods based on the results of a questionnaire.Pain at the time of insertion of an endoscope into the nasal cavity was common in all patients. It was experienced in 58.3%, 53.7% and 41.0% of patients treated by the spray, stick and stick + pharyngeal anesthesia methods, respectively. The pain was alleviated by the stick + pharyngeal anesthesia method significantly (p=0.022). Patients who did not experience any pain at the time of the endoscopic examination were 15.0%, 16.2% and 18.0%, respectively, with no significant differences.The least painful pretreatment procedure in the endoscopic examination was the stick + pharyngeal anesthesia method.

Capsule endoscopy (CE) represents a significant advance in the investigation of small bowel diseases. Little is known about the clinical outcome of patients with obscure gastrointestinal bleeding (OGIB).Seventy-eight patients underwent CE for OGIB and were followed up for at least 6 months after CE. The diagnostic yield of CE and the rate of re-bleeding during the follow-up period were established.Out of our 78 OGIB patients, 35 (44.9%) had significant lesions. There was a significant difference in the rate of identification of significant lesions between the on-going overt bleeding cases and previous overt bleeding cases (68.8% vs. 37.8%, respectively, p=0.043). Of the 46 patients with significant or insignificant lesions, 12 (26.1%) had one or more re-bleeding episodes during the follow-up period. On the other hand, only one (4%) of the 26 patients with negative findings had a re-bleeding episode (p=0.025).In conclusion, our study confirmed the role of CE in the diagnosis of OGIB, especially in the on-going overt bleeding cases. The OGIB patients with negative CE findings showed a low re-bleeding rate in the follow-up period. Further long-term follow-up studies are needed in future to examine the negative CE cases.

Infection with hepatitis C virus (HCV) is the leading cause of liver cirrhosis that develops into hepatocellular carcinoma. Previous studies have shown in vitro that lipids within hepatocytes are crucially important for a series of HCV infection-proliferation-release processes. On the other hand, in the patients with HCV, the serum total cholesterol (Total-C) and low-density lipoprotein cholesterol (LDL-C) levels have been reported to be lower. We conducted an epidemiological survey of a large cohort and investigated whether the lower serum lipid levels were caused by a direct or the secondary effects of HCV infection (i.e. hepatic damage or nutritional disorder). Among 146 857 participants (male, 34%; female, 66%) undergoing public health examinations between 2002 and 2007 in Ibaraki Prefecture, Japan, the HCV positive rates determined by HCV antibody/antigen and/or RNA tests were 1.37% and 0.67% in males and females, respectively. In addition to Total-C and LDL-C, serum high-density lipoprotein cholesterol and triglyceride concentrations were also significantly lower in the HCV positive subjects compared with the negative subjects, regardless of sex, age or nutritional state evaluated by body mass index. Multivariate analysis showed that HCV infection was the strongest among the factors to be significantly associated with the lower level of these lipids. Particularly, the hypolipidemia was also confirmed in the HCV positive subjects with normal aminotransferase levels (alanine aminotransferase ≤30 and aspartate aminotransferase ≤30). This epidemiological survey in a large Japanese cohort suggests that the HCV infection itself might directly cause hypolipidemia, irrespective of host factors including age, hepatic damage and nutritional state.

Although video capsule endoscopy (CE) is now an established modality for examining the small bowel, in almost 20 - 50 % cases, the capsule does not reach the colon by the end of the recording time. The aim of this study was to investigate whether metoclopramide increases the completion rate of CE examination.One hundred patients who underwent CE were classified to receive either intravenous treatment with metoclopramide (metoclopramide group: 43 cases) or no treatment at all (control group: 57 cases).GTT was 18.9 +/- 21.6 min in the metoclopramide group and 37.4 +/- 45.3 min in the control group (p = 0.0053). The capsule reached the cecum in 31 cases (75.6%) of the metoclopramide group and in 38 cases (69.1%) of the control group (p = 0.482). The SBTT and the combined GTT and SBTT were 276.8 +/- 108.9 min, 293.0 +/- 109.4 min in the metoclopramide group and 305.5 +/- 86.9 min, 339.4 +/- 89.2 min in the control group (p = 0.122, p = 0.035).Although 10 mg of metoclopramide with intravenous drip had shortened the GTT and the combined GTT and SBTT, there was no improvement in the rate of complete small bowel examination and the results of diagnostic yield. Further investigations are necessary to evaluate the concomitant use of other prokinetics or preparations to improve the rate of complete small bowel examination.