CYP1A2 Downregulation by Obeticholic Acid: Usefulness as a Positive Control for the In Vitro Evaluation of Drug-Drug Interactions.

Cytochrome P450 (CYP) downregulation is a mechanism of drug-drug interaction encountered in pharmaceutical development which is difficult to evaluate in vitro because of the scarcity of evidence. A previous clinical study of obeticholic acid (OCA) with caffeine suggested that OCA may be a useful positive control to establish a method to evaluate CYP1A2 downregulation and to investigate the mechanism of its downregulation. In the present study, we investigated the ability of OCA to downregulate CYP1A2 in human hepatocytes. OCA suppressed CYP1A2 mRNA expression and CYP1A2 enzyme activity without causing direct inhibition of CYP1A2 or cytotoxicity, suggesting that OCA downregulates CYP1A2 in vitro. OCA significantly suppressed the induction of CYP1A2 mRNA expression by omeprazole in a concentration-dependent manner, suggesting that a combination of inducers and new chemical entities would be helpful to investigate the mechanism of CYP1A2 downregulation and to evaluate the potential of new chemical entities for downregulation and investigate their downregulation mechanism. We also showed that CYP1A2 was transcriptionally downregulated by OCA and that a reduction in aryl hydrocarbon receptor mRNA expression is a possible mechanism of CYP1A2 downregulation by OCA. These results indicate that OCA would be a suitable positive control for studies of CYP1A2 downregulation.

A case of acute kidney injury and disseminated intravascular coagulation associated with influenza B viral infection.

There are few reports of acute kidney injury (AKI) associated with influenza viral infection. We treated a case of AKI that developed after an influenza B viral infection. A 35-year-old man visited a local physician for a fever and was diagnosed with influenza B. He was prescribed laninamivir, then returned to the physician 5 days later with dyspnea and was referred to Hospital A. Upon admission, respiratory arrest developed, for which he received tracheal intubation and mechanical ventilation. AKI was noted after admission and the patient was transferred to our hospital the next day. AKI and disseminated intravascular coagulation (DIC) were present at the time of transfer, thus a transfusion and continuous hemodiafiltration (CHDF) were performed, and administrations of thrombomodulin alpha and antithrombin III were initiated. Although the patient had DIC, AKI, and disturbance of consciousness, and was in a clinical state resembling influenza-associated encephalopathy, there was no clear abnormality shown in CT scans of the head. Urine output, renal function, and respiratory condition gradually improved, thus CHDF was stopped and extubation performed. The patient had no complications and was discharged on hospital day 22. Some reports have been presented regarding cases of AKI due to rhabdomyolysis associated with influenza viral infection, whereas our patient developed AKI as a complication of an influenza B viral infection without rhabdomyolysis or hemolytic uremic syndrome. Influenza B may cause AKI and DIC, and affected patients can be in a serious condition requiring immediate attention.

Association between gene polymorphisms of connective tissue growth factor and the progression of chronic liver disease associated with hepatitis C.

OBJECTIVE: Fibrogenic cytokines, such as transforming growth factor-beta 1 play a central role in the progression of liver fibrosis. Recently, functional gene polymorphisms in these cytokines have been identified, and some reports have validated the presence of associations between these polymorphisms and disease progression. Connective tissue growth factor (CTGF) is a stimulating factor for fibroblast proliferation and matrix production. This study aimed to examine the relationship between CTGF gene polymorphisms and the progression of hepatitis C virus (HCV)-related chronic liver disease, as well as the incidence and prognosis of hepatocellular carcinoma (HCC). METHODS: A review was conducted among 235 HCV patients (117 patients with chronic hepatitis (CH) and 118 patients with liver cirrhosis (LC)). The CTGF gene polymorphism (rs6918698; -945 G/C) was identified according to the chimeric cycling probe method. The rate of liver fibrosis progression was measured using two liver fibrosis prediction formulas, the Forns index and the FibroIndex. All HCC patients were followed regularly every month. RESULTS: The frequency of the -945 C allele was higher among the LC patients than the CH patients. Regarding the rate of liver fibrosis progression over five years, C homozygotes tended to exhibit a faster rate than G carriers, although the difference was not significant. Among the LC patients, the C homozygotes demonstrated lower prothrombin times, higher rates of indocyanine green retention and higher Child-Pugh scores than the G carriers. There were no significant tendencies in the genotype distribution, irrespective of the status of HCC. However, the prognosis of HCC was poorer for the C homozygotes than for the G carriers. CONCLUSION: A CTGF -945 C homozygote status is a significant risk factor for the progression of HCV-related chronic liver disease, including HCC.

Association of TIMP-1 and TIMP-2 gene polymorphisms with progression of liver fibrosis in patients with type C chronic liver disease.

We examined the association of TIMP-1 and TIMP-2 gene polymorphisms with the progression of chronic liver disease related to the hepatitis C virus (HCV). We used PCR to analyze 188 patients with HCV-related liver disease (95 with chronic hepatitis and 93 with cirrhosis) for TIMP-1 372 T/C and TIMP-2 -418 G/C polymorphisms. Comparing chronic hepatitis and cirrhosis, there were no significant differences in TIMP-1 and TIMP-2 gene polymorphisms. Among chronic hepatitis patients, TIMP-2 -418 G homozygotes showed significantly faster fibrosis progression than C carriers. Among cirrhotic patients, males with the TIMP-1 372 T allele developed cirrhosis at a younger age, and patients who were homozygous for the higher-transcription TIMP-2 -418 G allele had significantly lower serum albumin concentrations. These results suggest that faster progression of liver fibrosis could be associated with TIMP-2 -418 G homozygotes.

Drag current reversal in capacitively coupled arrays of small Josephson junctions.

The current-voltage characteristics of two long capacitively coupled one-dimensional arrays of small Josephson junctions were measured, where the coupling capacitance was comparable with the capacitance of the junction. The current in both arrays was simultaneously measured at various applied magnetic fields with the voltage applied to only one of the arrays. The current was induced in the unbiased array beyond the threshold voltage for the Coulomb blockade of the biased array. The direction of the induced current is the same as that of the current in the other array when the applied magnetic field is small and Cooper-pair tunneling is dominant, while it changes to the opposite direction when the applied magnetic field is large and single-electron tunneling is dominant. This suggests that the current induction mechanism between arrays of small Josephson junctions is substantially different from that in arrays of small normal tunnel junctions.

Divergent synthesis and evaluation of inhibitory activities against cyclooxygenases-1 and -2 of natural withasomnines and analogues.

The divergent synthesis of natural withasomnines and analogues was achieved from 4-hydroxypyrazoles, which was prepared via alkaline hydrolysis of the Baeyer-Villiger oxidation products from 4-formylpyrazoles. Key steps of this synthesis are regioselective Claisen rearrangement of 4-allyloxypyrazoles and the Suzuki-Miyaura coupling of 5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-yl trifluoromethanesulfonate and commercially available arylboronic acids. The Suzuki-Miyaura coupling at the final step of this strategy enabled facile access to natural withasomnines and their analogues. The biological activities of the twelve synthesized compounds against cyclooxygenases-1 and -2 (COX-1 and COX-2) were evaluated.

Functional gene polymorphisms of interleukin-10 are associated with liver disease progression in Japanese patients with hepatitis C virus infection.

OBJECTIVE: The inter-individual difference in response to liver injury appears to be important in the progression of liver fibrosis. Interleukin 10 (IL-10) is an anti-inflammatory cytokine, and several functional gene polymorphisms have been found. The aim of this study was to examine the possible association of IL-10 polymorphisms with the progression of liver fibrosis in hepatitis C virus (HCV)-related chronic liver disease patients. METHODS: We examined the IL-10 -1087 A/G and -824 T/C gene polymorphisms in 184 Japanese patients with HCV-related chronic liver disease: 94 chronic hepatitis (CH) and 90 with liver cirrhosis (LC). RESULTS: There were no significant differences in the genotype distributions or allele frequencies of IL-10 -824 T/C and -1087 A/G between the CH and LC groups. However, among the cirrhotic patients, the lower transcriptional allele, -824 T homozygotes had significantly lower serum albumin and platelet counts, and a higher Child-Pugh score than the -824 C carriers, and the lower transcriptional allele, -1087 A homozygotes had a higher ICG-R 15 compared with -1087 G carriers. Haplotype analysis of IL-10 -1087/-824 showed no significant difference between the CH and LC groups, but the combinations of AT and AC haplotypes (AT/AT, AT/AC and AC/AC) had a significantly higher ICG-R 15 than the GC carriers. CONCLUSION: IL-10 lower transcriptional -824 T allele, -1087 A allele, and -1087/-824 haplotypes AT and AC are risk factors for the progression of liver fibrosis in HCV-related chronic liver disease.

The Effects of Olmesartan and Alfacalcidol on Renoprotection and klotho Gene Expression in 5/6 Nephrectomized Spontaneously Hypertensive Rats.

Recently, an angiotensin inhibitor has been shown to upregulate the klotho mRNA level in chronic renal failure. In addition, the administration of vitamin D has been reported to improve the mortality of patients with chronic renal failure. In this study, we examined the effects of an angiotensin inhibitor and/or vitamin D on the progression of chronic renal failure by using male 5/6 nephrectomized (5/6Nx) spontaneously hypertensive rats. Male 5/6Nx spontaneously hypertensive rats were assigned to 4 groups as follows: 5/6Nx group, 5/6Nx rats; Alf group, 5/6Nx rats administered alfacalcidol (0.2 µg/kg/day); Olm group, 5/6Nx rats administered olmesartan (15 mg/kg/day); Alf + Olm group, 5/6Nx rats administered alfacalcidol (0.2 µg/kg/day) and olmesartan (15 mg/kg/day). These drugs were administered for 12 weeks. Systolic blood pressure in the Alf, Olm and Alf + Olm groups were significantly decreased relative to that in the 5/6Nx group during the 12-week experimental period. As a result, all treated groups showed renoprotection based on improvement of the systolic blood pressure, urinary protein excretion and histological renal fibrosis. Combination therapy of alfacalcidol and olmesartan was more effective than either alfacalcidol or olmesartan alone. Expression of klotho mRNA was significantly upregulated in the Alf + Olm group in comparison with in the 5/6Nx group. Serum levels of fibroblast growth factor 23 in the Alf group and the Alf + Olm group were significantly higher than those in the 5/6Nx group and the Olm group. In conclusion, the combination of Olm and Alf inhibited the progression of renal damage in the 5/6Nx group through the strong antihypertensive effect as well as the upregulation of the klotho gene.

The genotypes of IL-1 beta and MMP-3 are associated with the prognosis of HCV-related hepatocellular carcinoma.

BACKGROUND AND AIM: Cytokines and matrix metalloproteinases (MMPs) are involved in tumor growth, invasion, and remote metastasis in various cancers. Recently, functional gene polymorphisms in these cytokines and MMPs have been found, and some reports have revealed an association between these polymorphisms and the prognosis of various cancers. In this study, we examined the relationship between the gene polymorphisms of interleukin 1 beta (IL-1b), IL-1 receptor antagonist (IL-1 RN), transforming growth factor beta 1 (TGF-b1), MMP-1, MMP-3, and MMP-9 and the prognosis of hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC). METHODS: We enrolled 92 HCV-related HCC patients in the study, and gene polymorphisms of IL-1b -31 C/T, IL-1 RN variable number of tandem repeats (VNTR), TGF-b1 +869 C/T, MMP-1 -1,607 1G/2G, MMP-3 -1,171 5A/6A, and MMP-9 -1,562 C/T were analyzed. RESULTS: In HCC clinical features, TGF-b1 C carriers and MMP-3 5A carriers had significantly larger HCC diameters than TGF-b1 T and MMP-3 6A homozygotes. In HCC prognosis, IL-1b T homozygotes and MMP-3 5A carriers had a significantly poorer prognosis than IL-1b C carriers and MMP-3 6A homozygotes. Those with a combination of IL-1b T homozygosity and MMP-3 5A had synergistically poorer HCC prognosis. CONCLUSION: The IL-1b -31 T allele and MMP-3 5A allele are cooperative risk factors for poor prognosis in HCC patients, suggesting that these gene polymorphisms might be potential markers for predicting the prognosis of HCC patients.