Regulation of bile acid metabolism in mouse models with hydrophobic bile acid composition.

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.

Current states of prevention of drug-induced gastroduodenal ulcer in real clinical practice: a cross-sectional study.

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<0.05), but then started to decrease in 4th period; and the percentage of all patients taking NSAIDs and LDA decreased significantly (p<0.05) until 5th period. Among the 292 patients with gastroduodenal ulcer and the 121 patients with a bleeding ulcer taking NSAIDs and LDA, 16 (5.5%) and 9 (7.4%), 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. In 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 suggests the need to enlighten physicians about preventive treatment because drug withdrawal of LDA has a high risk of cardiovasculr and cerebrovascular events.

Anticholestatic effects of bezafibrate in patients with primary biliary cirrhosis treated with ursodeoxycholic acid.

UNLABELLED: 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 4ß-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.

Population pharmacokinetics of tazobactam/piperacillin in Japanese patients with community-acquired pneumonia.

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.

Sex-, age-, and organ-dependent improvement of bile acid hydrophobicity by ursodeoxycholic acid treatment: A study using a mouse model with human-like bile acid composition.

Cyp2a12-/-Cyp2c70-/- double knockout (DKO) mice have a human-like hydrophobic bile acid (BA) composition and show reduced fertility and liver injury. Ursodeoxycholic acid (UDCA) is a hydrophilic and cytoprotective BA used to treat various liver injuries in humans. This study investigated the effects of orally administered UDCA on fertility and liver injury in DKO mice. UDCA treatment prevented abnormal delivery (miscarriage and preterm birth) in pregnant DKO mice, presumably by increasing the hydrophilicity of serum BAs. UDCA also prevented liver damage in six-week-old DKO mice, however liver injury emerged in UDCA-treated 20-week-old female, but not male, DKO mice. In 20-week-old male UDCA-treated DKO mice, conjugated plus unconjugated UDCA proportions in serum, liver, and bile were 71, 64, and 71% of the total BAs, respectively. In contrast, conjugated plus unconjugated UDCA proportions in serum, liver, and bile of females were 56, 34, and 58% of the total BAs, respectively. The UDCA proportion was considerably low in female liver only and was compensated by highly hydrophobic lithocholic acid (LCA). Therefore, UDCA treatment markedly reduced the BA hydrophobicity index in the male liver but not in females. This appears to be why UDCA treatment causes liver injury in 20-week-old female mice. To explore the cause of LCA accumulation in the female liver, we evaluated the hepatic activity of CYP3A11 and SULT2A1, which metabolize LCAs to more hydrophilic BAs. However, there was no evidence to suggest that either enzyme activity was lower in females than in males. As female mice have a larger BA pool than males, excessive loading of LCAs on the hepatic bile salt export pump (BSEP) may be the reason for the hepatic accumulation of LCAs in female DKO mice with prolonged UDCA treatment. Our results suggest that the improvement of BA hydrophobicity in DKO mice by UDCA administration is sex-, age-, and organ-dependent.

Evaluation of gut dysbiosis using serum and fecal bile acid profiles.

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 Clostridium difficile infection (CDI). Since XIVa is a bacterial group responsible for the conversion of primary bile acids (BAs) to secondary BAs, the proportion of intestinal XIVa can be predicted by determining the ratio of deoxycholic acid (DCA)/[DCA + cholic acid (CA)] in feces orserum. For example, serum DCA/(DCA+CA) was significantly lower in IBD patients than in healthy controls, even in the remission period. These results suggest that a low proportion of intestinal XIVa in IBD patients might be a precondition for IBD onset but not a consequence of intestinal inflammation. Another report showed that a reduced serum DCA/(DCA + CA) ratio could predict susceptibility to CDI. Thus, the BA profile, particularly the ratio of secondary to primary BAs, can serve as a surrogate marker of the intestinal dysbiosis caused by decreased XIVa.

Cholesterol 25-hydroxylation activity of CYP3A.

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 4ß-hydroxylation, a known marker activity of CYP3A4. In addition, 25-hydroxycholesterol concentrations in normal human sera correlated positively with the levels of 4ß-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.

Pretreatment methods in transnasal endoscopy.

BACKGROUND/AIMS: 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. METHODOLOGY: 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. RESULTS: 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. CONCLUSION: The least painful pretreatment procedure in the endoscopic examination was the stick + pharyngeal anesthesia method.

The clinical outcome of capsule endoscopy in patients with obscure gastrointestinal bleeding.

BACKGROUND/AIMS: 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). METHODOLOGY: 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. RESULTS: 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). CONCLUSION: 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.

Dose conversion in opioid rotation from continuous intravenous infusion of morphine hydrochloride injection to fentanyl patch in the management of cancer pain.

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.

Western Diet Changes Gut Microbiota and Ameliorates Liver Injury in a Mouse Model with Human-Like Bile Acid Composition.

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.

Clinical features of gastroduodenal ulcer in Japanese patients taking low-dose aspirin.

BACKGROUND AND AIM: The risks of peptic ulcer complications increase in association with low-dose aspirin (LDA) use. The endoscopic findings and clinical features of gastroduodenal ulcer have not been thoroughly investigated in patients taking LDA. METHOD: We classified 1,041 gastroduodenal ulcer patients into three groups [patients taking LDA (group A), patients taking nonaspirin nonsteroidal anti-inflammatory drug (NSAID) (group N), and patients taking neither aspirin nor nonaspirin NSAID (group C)] and 241 bleeding gastroduodenal ulcer patients into three corresponding groups (groups a, n, and c). We investigated the clinical features, endoscopic characteristics, and endoscopic treatment of the hemorrhagic lesion in the gastroduodenal ulcer patients taking LDA and compared them with those of the other groups. RESULTS: The frequency of bleeding events such as hematemesis, melena, and anemia was significantly higher in group A and N than in group C. The percentage of ulcers located in the antrum was higher in group A and N than in group C, and also higher in group a and n than in group c. The percentage of ulcers located in the body, fundus, and cardia was significantly higher in the bleeding patients than in all gastroduodenal ulcer patients. The percentage of cases that required additional endoscopic treatment in group a was higher than in group c. Duration of hospitalization of group a was significantly longer than that of group c. CONCLUSION: These results indicate that it is very important to prevent LDA-induced gastroduodenal ulcer complications, including bleeding.

Clinicopathological features of undifferentiated mixed type early gastric cancer treated with endoscopic submucosal dissection.

BACKGROUND/AIMS: Previous published data on the differentiated/undifferentiated mixed-type early gastric cancer treated with ESD are scarce. METHODOLOGY: The clinicopathological features of the pure differentiated type and differentiated/undifferentiated mixed type adenocarcinoma in final diagnosis of ESD specimens were investigated in 109 early gastric cancers patients treated with ESD. RESULTS: Of the pure differentiated type (102 patients) and differentiated / undifferentiated mixed type (7 patients) adenocarcinoma, submucosal invasive tumors were detected in 6 (5.9%) and 4 cases (57.1%) (p = 0.017), respectively. The mean tumor size was 10.95 +/- 5.12mm and 22.85 +/- 9.51mm (p = 0.0091), respectively. In the cases diagnosed as moderately differentiated adenocarcinoma by preoperative ESD biopsy, submucosal invasive tumors were detected in 2 (12.5%) and 4 cases (57.1%) (p = 0.038), respectively, and the mean tumor size was 12.50 +/- 5.88mm and 22.85 +/- 9.51mm (p = 0.016), respectively. During a median follow-up period of 30.3 months (range: 7-52 months), there were no recurrences or metastasis in any of the 7 mixed type cases. CONCLUSION: In conclusion, the results of our study suggest that large gastric tumors treated with ESD which are diagnosed as moderately differentiated adenocarcinoma in biopsy specimens potentially contain an undifferentiated component.

The effect of metoclopramide in capsule endoscopy.

BACKGROUND/AIMS: 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. METHODOLOGY: 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). RESULTS: 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). CONCLUSION: 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.

Potential involvement of the stem cell factor receptor c-kit in alopecia areata and androgenetic alopecia: histopathological, immunohistochemical, and semiquantitative investigations.

Alopecia areata (AAR) and androgenetic alopecia (AGA) are two major forms of alopecia based on altered hair growth condition. In general, the cell cycle is regulated by several mechanisms including the stem cell factor/c-kit signaling. To assess a role for stem cell activity in alopecia, we performed histopathological, immunohistochemical, and semiquantitative analyses of c-kit as well as Ki-67 in scalp biopsy specimens obtained from 14 patients with AAR, 18 patients with AGA, and 6 age-matched control subjects, using the specific antibodies. Formalin-fixed, paraffin-embedded skin sections were examined. Immunoreactivities for Ki-67 and c-kit were localized in keratinocytes and melanocytes in the outermost layer of hair follicles. The mean length of hair follicles was significantly shorter in the AAR and AGA groups than in the control group. The mean number of Ki-67-immunoreactive cells per follicle was significantly reduced in the AAR and AGA groups as compared with the control group. The mean number of c-kit-immunoreactive cells per follicle was significantly increased in the AAR and AGA groups as compared with the control group. Our results indicate that c-kit is upregulated in the hair follicle cells in these forms of alopecia, and suggest that the upregulation reflects a negative feedback mechanism in response to possible downregulation of the ligand stem cell factor.

[Liver Injury Risk Factors in Amyotrophic Lateral Sclerosis Patients Treated with Riluzole].

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.

Highly sensitive quantification of serum malonate, a possible marker for de novo lipogenesis, by LC-ESI-MS/MS.

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.

Serum concentration of 27-hydroxycholesterol predicts the effects of high-cholesterol diet on plasma LDL cholesterol level.

AIM: The effect of dietary cholesterol on plasma cholesterol concentrations varies widely among individuals. Recent studies suggest that the synthesis of oxysterols is up-regulated when tissue cholesterol is saturated. The present study was undertaken to test the hypothesis that a serum high concentration of 27-hydroxycholesterol, one of the oxysterols, reflects positive cholesterol balance in the body and predicts intolerance to a high-cholesterol diet. METHODS: In 30 subjects, 750 mg/day of cholesterol was added for 4 weeks to the ordinary diet. Blood samples were collected at the start and finish of the supplementation. Serum sterol and oxysterol concentrations were measured by high-resolution GC-MS. RESULTS: A receiver operating characteristic curve was drawn and the cutoff point (80 ng/mg cholesterol) was chosen to maximize sensitivity (81.3%) and specificity (64.3%) for predicting a positive change of LDL cholesterol concentration after cholesterol loading. Subjects with higher serum 27-hydroxycholesterol concentrations (>/= 80 ng/mg cholesterol) showed significantly (P < 0.05) high values for the change of LDL cholesterol concentration (+7.4 +/- 3.4%, mean +/- SEM, n = 17) compared with those with lower 27-hydroxycholesterol levels (-5.3 +/- 2.7%, n = 13). CONCLUSIONS: In subjects with high serum 27-hydroxycholesterol concentrations were unable to adapt to a high-cholesterol diet. The concentration of serum 27-hydroxycholesterol appears to reflect cholesterol saturation in the body and predicts to some extent a responsiveness to dietary cholesterol.

The associated markers and their limitations for the primary screening of HCV carriers in public health examination.

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.

Impact of determination of hepatitis B virus subgenotype and pre-core/core-promoter mutation for the prediction of acute exacerbation of asymptomatic carriers.

AIM: A large cohort study in Japan revealed that the specific viral profile may influence the fulminant outcome in acute hepatitis B virus (HBV) infections, while the genetic influence on outcome has not been clarified in patients with acute exacerbation of chronic liver disease caused by HBV. We experienced a case of fatal liver failure that developed as the result of chronic HBV infection. To determine possible genetic factor involving acute exacerbation, genetic analysis of serum from the patient and his siblings was performed. METHODS: HBV subgenotype as well as pre-core/core-promoter mutations of samples mentioned above were determined. RESULTS: Patient had HBV-Bj with pre-core (1896/1899) and core-promoter (1762/1764) mutations, the genomic profile frequently seen in fulminant hepatitis caused by acute HBV infection. CONCLUSION: This result suggests that determination of the HBV subgenotype and pre-core/core promoter mutations could provide a rationale for development of a treatment strategy in asymptomatic HBV carriers.