Short interfering RNA-directed inhibition of hepatitis B virus replication

AbstractRNA interference (RNAi) is the process by which double-stranded RNA directs sequence-specific degradation of mRNA. In mammalian cells, RNAi can be triggered by 21-nucleotide duplexes of short interfering RNA (siRNA). We examined effects of siRNA on hepatitis B virus (HBV) replication. Human hepatoma cells were transfected with HBV DNA and siRNA against HBV-pregenome RNA. Transfection experiments demonstrated that the siRNA reduced the amount of HBV-pregenome RNA and resulted in reduction of the levels of replicative intermediates and viral protein. Our results indicate that siRNA-mediated gene silencing inhibits HBV replication through suppression of viral RNA, which may be useful as a potential therapeutic modality

Gas-Liquid Interfacial Area, Bubble Size and Liquid-Phase Mass Transfer Coefficient in a Three-Phase External Loop Airlift Bubble Column

The interfacial area a was measured by the sulfite oxidation method in a three-phase external loop airlift bubble column suspending completely the different concentrations of ion exchange resin particles in aqueous carboxymethyl cellulose (CMC) solutions with a wide range of viscosity. The column had been previously studied for the circulating liquid velocity UL, gas holdup G and volumetric gas-liquid oxygen transfer coefficient kLa in the two- and three-phase systems. The average bubble size dB and oxygen transfer coefficient kL were obtained as dB = 6 G/a and kL = (the previous kLa)/a, respectively. The similar studies were carried out in the internal loop airlift and normal bubble columns for comparison. The a values in the external loop airlift were found to be little affected by the column height and particles concentrations, and to decrease with increasing viscosity. All the three columns showed a linear dependence of a on G. A simple correlation of a, dB or kL was proposed as a function of G and viscosity for the external loop airlift as well as both internal loop airlift and normal columns. A well-known relationship between kL and dB was confirmed to hold independent of column types and operating conditions for a given two or three phase system

Cloning of cDNA and genomic DNA for human cytochrome P-45011β

AbstractA full-length cDNA clone encoding steroid 11β-hydroxylase (P-45011β) has been isolated from a cDNA library derived from human adrenal tumor. The insert of the clone contains an open reading frame encoding a protein of 503 amino acid residues together with a 4 bp 5'-untranslated region and a 576 bp 3'-untranslated region to which a poly(A) tract is attached. The promoter region of the P-45011β gene has also been isolated from a genomic library derived from human pre-B cells. It contains a TATA box, a putative cAMP-responsive element, several repeated sequences and two sequence elements similar to the consensus sequence for binding of AP-1. A transient expression assay in Y-1 adrenal tumor cells demonstrates that the promoter activity is remarkably enhanced by treatment of the cells with cAMP. In addition, analysis using deletion mutants containing various lengths of the 5'-flanking region of the gene suggests that several cis-acting elements participate in transcriptional regulation of human P-45011β gene

A Case of Pancreatic Ascites and Pleural Effusion: Confirmation of a Pancreatic Duct Contrast Leakage Using Computed Tomography after Endoscopic Retrograde Cholangiopancreatography

A seventy-two year old Japanese man with chronic alcoholism was admitted with increasing epigastric pain and abdominal fullness. He gave a history of bouts of epigastric pain radiating to the back for the past year. At admission, abdominal ultrasonography and computed tomography (CT) demonstrated massive ascites and a pseudocyst in the pancreatic body. A chest X-ray showed bilateral pleural effusion, and the level of amylase was elevated in the serum, urine, ascitic fluid and pleural effusion. First, the patient was treated with nothing by mouth but with intravenous hyperalimentation, however, no improvement was noted after 2 weeks. Then, the patient underwent endoscopic retrograde cholangiopancreatography (ERCP) and abdominal CT after ERCP. They showed irregular dilatation of the pancreatic main duct and branch, and an extravasation of contrast media from the pancreatic duct into the peritoneal cavity, after which the patient underwent surgery. Because no fistula was found during surgery, drainages were retained into the pseudocyst and peritoneal cavity. Due to marked elevation of amylase and protein levels in ascitic fluid and pleural effusion and findings from ERCP and CT after ERCP, pancreatic ascites and pleural effusion was diagnosed. The diagnosis of chronic pancreatitis is due to his history, laboratory data, and irregular dilatation of the pancreatic duct on ERCP. After surgery, his clinical status improved rapidly. We thus described a case of pancreaticoperitoneal fistula demonstrated by CT scan subsequent to ERCP which was treated successfully by surgery

Deficiency of the RIβ subunit of protein kinase A causes body tremor and impaired fear conditioning memory in rats

The RIβ subunit of cAMP-dependent protein kinase (PKA), encoded by Prkar1b, is a neuronal isoform of the type I regulatory subunit of PKA. Mice lacking the RIβ subunit exhibit normal long-term potentiation (LTP) in the Schaffer collateral pathway of the hippocampus and normal behavior in the open-field and fear conditioning tests. Here, we combined genetic, electrophysiological, and behavioral approaches to demonstrate that the RIβ subunit was involved in body tremor, LTP in the Schaffer collateral pathway, and fear conditioning memory in rats. Genetic analysis of WTC-furue, a mutant strain with spontaneous tremors, revealed a deletion in the Prkar1b gene of the WTC-furue genome. Prkar1b-deficient rats created by the CRISPR/Cas9 system exhibited body tremor. Hippocampal slices from mutant rats showed deficient LTP in the Schaffer collateral–CA1 synapse. Mutant rats also exhibited decreased freezing time following contextual and cued fear conditioning, as well as increased exploratory behavior in the open field. These findings indicate the roles of the RIβ subunit in tremor pathogenesis and contextual and cued fear memory, and suggest that the hippocampal and amygdala roles of this subunit differ between mice and rats and that rats are therefore beneficial for exploring RIβ function

Comparative study of the work load between one-man buses and two-man buses.

The differences in physiological and safety conditions of one-man buses and two-man buses were examined from the view point of occupational fatigue. This survey consisted of a work load study which included a time study, study of subsidiary behavior, auditory task, memory test, Galvanic Skin Response (GSR) and physiological function tests and a self-administered questionnaire which involved items concerning safety and subjective fatigue complaints. The visual and postural restrictions in the one-man bus were greater than in the two-man bus. The mental capacity of the one-man bus drivers was found to be less. Greater mental fatigue and stress were observed in the one-man bus. More subjective fatigue complaints were observed in the one-man bus. More cases of near accidents were observed in the one-man bus. From these results it was concluded that the one-man bus caused bus drivers a greater mental and physical work load.</p

The role of IL-18 in the modulation of matrix metalloproteinases and migration of human natural killer (NK) cells

AbstractIn this study, we examined whether interleukin-18 (IL-18) affects natural killer (NK) cells' migration and matrix metalloproteinases (MMPs) production. We demonstrated that chemotaxis of human NK cells through basement membrane-like Matrigel was augmented by IL-18. As well, IL-18 stimulation induces the production of activated forms of matrix metalloproteinase-2 (MMP-2) as well as the production of pro-MMP-2 from NK cells. We also demonstrated that MT1-MMP expression on human NK cells, which is a major activator of MMP-2, was induced by IL-18 stimulation coordinated with MMP-2 activation. These data suggest that the MT1-MMP/MMP-2 system participates in the degradation of basement membrane components and thus contributes to NK cell migration

Attenuation of Responsiveness to Interferon-α Treatment by Preceded Overactivation of Interferon-mediated Pathway in Patients Chronically Infected by Hepatitis C Virus

The Interferon (IFN) receptor-mediated signal transduction pathways involve the two novel DNA-binding factors, interferon regulatory factor-1 (IRF-1) and IRF-2. Both recognize the same DNA sequence, in which the expression ratio of IRF-1 to IRF-2 is relevant to induction of IFN-inducible genes, because IRF-1 acts as a transcriptional activator and IRF-2 as a counterpart. In the present study, 54 patients with chronic hepatitis C and the age-and sex-matched 7 subjects with fatty liver as a control were subjected to analysis of the expression ratio of IRF-1 to IRF-2 mRNA in the liver tissue obtained at the time of livert biopsy by reverse transcription-polymerase chain reaction in combination with a restriction fragment length polymorphism assay. The expression ratio of IRF-1 to IRF-2 mRNA in the liver tissue in patients chronically infected by hepatitis C virus (HCV) was significantly higher than that in control, although the values did not correlate with the serum levels of HCV-RNA. Of 54 patients, 28 received IFN treatment, resulting in complete response in 8 patients. With respect to responsiveness to IFN treatment, patients who had complete response had the relatively lower ratios of IRF-1 to IRF-2 mRNA in the liver tissue, compared with those who did not. These results indicate that the IFN-mediated pathway is spontaneously activated in patients with chronic hepatitis C, and that its preceded overactivation counteracts on the efficacy of IFN treatment in these patients

Macrophage-dominant sialadenitis in human T-cell leukemia virus type I-associated myelopathy after living-donor liver transplantation.

A 64-year-old man who suffered from human T-cell leukemia virus type I (HTLV-I)-associated myelopathy (HAM) after living-donor liver transplantation (LDLT) for liver cirrhosis due to hepatitis C virus infection complained of xerostomia. Although exocrine function test results were positive, autoantibodies including anti-SS-A/SS-B antibodies and sialography showed negative findings. Labial salivary gland biopsy revealing infiltration of 60 counts of mononuclear cells (MNCs) in minor salivary glands led to a diagnosis of Sjögren\u27s syndrome-like sialadenitis. Immunohistochemistry demonstrated dominant CD68 staining and major histocompatibility complex class II on the surface of infiltrating MNCs. Herein we have reported a rare condition of macrophage-dominant sialadenitis in a patient with HAM after LDLT