Search

The claudin family comprises four-pass transmembrane proteins involved in the formation of tight junctions (TJs). Relatively recently, claudin domain containing (CLDND) 1, also known as claudin-25, was identified as a novel member of the claudin family. In the present study, we revealed that in the adult murine brain, CLDND1 is abundant in the cerebellum among common sites of intracerebral hemorrhage. Thus, the dynamics of CLDND1 after cerebellar hemorrhage were examined. Both CLDND1 mRNA and protein levels transiently decreased at 24 hr after hemorrhagic insult. For immunostaining, an anti-CLDND1 antibody that recognizes the specific epitope in the extracellular first loop was prepared. Dual immunohistochemical staining with CD31 using coronal cryosections of intact murine cerebellum tissue revealed that CLDND1 is expressed on endothelial cells. We therefore performed an in vitro permeability test using a human brain endothelial cell (HBEC) line to reveal whether CLDND1 contributes to cell adhesion like other claudins. CLDND1 was expressed on HBECs as well as in murine cerebellum tissue, and a strong signal was observed at TJs. RNA interference against CLDND1 decreased both the mRNA and protein levels without cytotoxicity. The permeability to small molecules, but not to large ones, across confluent HBECs increased on CLDND1 knockdown compared with mock-treated cells. These results suggest that the transient decrease of CLDND1 after cerebellar hemorrhage is responsible for low-molecular-weight selective vascular hyperpermeability. © 2017 Wiley Periodicals, Inc.

The wnt protein family has important members involved in cell differentiation, proliferation and plasticity expression; however, little is known about its biosynthesis processes. On the other hand, an increase in the intracerebral cyclic adenosine 3', 5'-monophosphate (cAMP) level leads to synaptic plasticity via the de novo synthesis of any protein. Here, the effect of dibutyryl cAMP (dbcAMP), a membrane permeability cAMP analog, on the wnt family was investigated in rat primary-cultured glial cells containing astrocytes and microglia. Among wnt3a, 4, 5a, 7a and 11 mRNA, only wnt4 expression was increased by longer treatment (24 h), compared with short treatment (2 h), with dbcAMP in a concentration-dependent manner, and its effect reached statistical significance at 1 mM. In cultures of isolated astrocytes or microglia, wnt4 expression was not affected by 1 mM dbcAMP for 24 h, and microglial wnt4 protein was undetectable even when cells were treated with the drug. Mixed glial cells treated for 24 h with 1 mM dbcAMP showed significantly increased wnt4 protein, as well as mRNA. Immunofluorescence manifested that cells that expressed wnt4 protein were astrocytes, but not microglia. Intraperitoneal injection of 1.25 mg/kg rolipram, a phosphodiesterase (PDE) IV inhibitor that can pass through the blood brain barrier and inhibits cAMP degradation specifically, showed a tendency to increase wnt4 expression in the adult rat brain after 24 h, and the increases in wnt4 mRNA and protein levels reached statistical significance in the hippocampus and striatum, respectively. This is the first finding to help elucidate the selective biosynthesis of central wnt4 through cAMP-stimulated microglia and astrocytes interaction.

The pharmacokinetics, distribution, and excretion of radioactivity and the main metabolites, M-1, M-2 and M-3, were studied in male rat after a single intravenous administration of 14C-labeled roxatidine acetate hydrochloride (TZU) at a dose of 10 mg/kg. The concentrations of these three metabolites in plasma, tissues and urine were simultaneously quantified with LC/MS-SIM using corresponding deuterium-labeled compounds as internal standards.1) The total radioactivity in the blood decreased with the half-lives of 0.38 and 3.82 hr in α-phase and β-phase, respectively. The plasma concentration of M-1, the active metabolite, decreased with the half-lives of 0.17 hr in α-phase and 0.77 hr in β-phase.2) At 15 min after administration, a high radioactivity was found in the liver, kidney, stomach, small intestine, parotid and submaxillary glands. At 72 hr after administration, the radioactivity was less than 0.1% of the dose or below the detection limit in any tissues.3) The metabolites in tissues were qualitatively analyzed by radio-HPLC. The radioactivity of the M-1 fraction was the highest in the brain, stomach, small intestine and kidney, then followed by that of the M-2 and M-3, the total radioactivity mainly consisted of those three fractions.4) The concentrations of three metabolites were quantified with LC/MS-SIM in the brain, lung, stomach, small intestine, liver, kidney and testis. All these metabolites disappeared rapidly, the half-lives were below 3.4 hr, in every tissue except for the testis. Although the radioactivity in the testis slowly decreased with the half-life of 10.5 hr, it accounted for M-2 and M-3, because they disappeared similarly to the radioactivity.5) In the target organs, stomach and small intestine, the concentration of M-1 was the highest, confirming that TZU has an advantage as an H2-receptor antagonist.6) Only in the liver, the level of the phenolic metabolite, M-3, was the highest of the three metabolites. This finding indicated that the metabolite might be mainly produced through oxidative dealkylation in liver.7) Approximately 87 and 8 % of the dosed radioactivity were excreted within 48 hr after administration in the urine and feces, respectively. The main metabolite in the urine was M-3, consisting of 29% of the dose, and about half of that was in the conjugated form. In feces, the excretion ratios of these three metabolites were almost equal, 0.3% of the dose.8) About 20% of the dosed radioactivity was excreted into bile in billiary fistular rats. The radioactivities in the M-1 and M-2 fractions were each 0.3% of the dose. The M-3 fraction comprised about 8%, almost all of which was in the conjugated form.

A 92-year-old woman with a brain tumor developed swelling of the left lower extremity. Venography showed considerable thrombi from the left common iliac vein to the femoral vein. Following implantation of a temporary inferior vena cava filter, catheter aspiration therapy and catheter-directed thrombolysis were performed. Venography after 3 days showed disappearance of the thrombi and an improvement in vein flow. A permanent inferior vena cava filter was implanted. Local intensive thrombectomy and thrombolysis by catheter together with a temporary inferior vena cava filter were effective treatments in this elderly patient with deep vein thrombosis.

An 86-year-old woman was admitted to our hospital because of vomiting and anorexia. Although serum cardiac markers, an electrocardiogram, and echocardiography suggested acute myocardial infarction, emergency cardiac catheterization revealed akinesis of the left ventricular apex without significant coronary artery stenosis. She was diagnosed as having takotsubo cardiomyopathy. The left ventricular dysfunction was considered transient and reversible but did not improve at all, contrary to our expectations. She died of worsening heart failure on day 14. We discuss this serious clinical course of a very elderly patient with takotsubo cardiomyopathy.

We study the structure of the twisted homology groups attached to weighted lines in the plane with resonant singular points, and find possible intersection pairings. As a typical example, we treat homology groups attached to 2-dimensional Selberg-type integrals.

1. 1. TZC-5665 is a novel pyridazinone derivative with vasodilatory and β-adrenergic blocking activities and type III phosphodiesterase inhibitory action. 2. 2. In healthy volunteers, TZC-5665 was rapidly absorbed and immediately metabolized. Its main metabolite, M-2, remained at a higher concentration in plasma. Orally administered TZC-5665 reduced end-diastolic left ventricular volume (20.16 ml) and exhibited a tendency to increase ejection fraction (0.04). 3. 3. In dogs, M-2 dose-dependently increased cardiac contractility and reduced both preload and afterload. These effects appeared more potent in the failed heart than in the normal heart. At the same dose (30 μg/kg), the effects of M-2 seem to be more potent than those of milrinone. 4. 4. We concluded that TZC-5665 is a useful medication for treating patients with chronic congestive heart failure (CHF) because of the positive inotropic and vasodilating effects due to its active metabolite in addition to its own β-adrenergic blocking actions.

State analysis of low-sulfated chondroitin 4-sulfate (LSC) in human urine and serum was performed by the use of high performance liquid chromatography and Western blot analysis. It was revealed that the most amount of LSC in urine is present as urinary trypsin inhibitor and a small amount (about 10% of total LSC) is as an LSC chain. The LSC in serum is mainly present as a proteoglycan such as inter-alpha-trypsin inhibitor (ITI), with a molecular weight of 212 kDa, but a small amount of LSC-proteoglycans having molecular weights of 128 and 38 kDa were also observed on SDS-PAGE. Those two compounds may be fragments of ITI, or one of the compounds (128 kDa) may be pre-alpha-trypsin inhibitor which was found by Enghild et al. (J. Biol. Chem., 264, 15975 (1989)).