Rat strain differences in the early stage of porcine-serum-induced hepatic fibrosis.

Rat strain differences in the early development of porcine serum (PS)-induced hepatic fibrosis were histologically and immunohistochemically examined using Brown Norway (BN), Sprague Dawley (SD) and Wistar rats. They were injected i.p. with 0.5 ml sterile PS twice a week for 4 and 8 weeks. In addition, rats treated with physiological saline in the same way served as controls. At 4 weeks, hepatic fibrosis accompanying fibrous septa mainly composed of type III collagens developed in BN and SD rats but not in Wistar rats. In addition, the numbers of eosinophils, CD3-positive cells and ED-1-positive cells significantly increased in BN and SD rats, that of CD45RA-positive cells in BN rats, and that of alpha-smooth muscle actin (SMA)-positive cells in SD rats, respectively. Such differences in the number of inflammatory cells may be related with the absence of hepatic fibrosis in Wistar rats at 4 weeks. At 8 weeks, hepatic fibrosis with formation of many small-sized pseudolobules was observed in all strains at almost similar degree, and the numbers of infiltrating cells increased in all strains of rats with some exception. In addition, the main location of inflammatory cells was different, suggesting a different role of each inflammatory cell in the process of hepatic fibrosis.     

Studies on formaldehyde resins, 19. General base catalysis in hydroxymethylation of melamine with formaldehyde

Hydroxymethylation of melamine with formaldehyde to form N-(hydroxymethyl)melamine (2,4-diamino-6-hydroxymethylamino-1,3,5-triazine) was investigated kinetically by the use of hydrogen phosphate/phosphate buffers in aqueous media at pH 11 ? 12. This reaction was found to follow a general base catalysis which results from the kinetic investigation, showing that the reaction takes place by a concerted mechanism involving base, melamine, and formaldehyde. This mechanism differs from that of the base catalyzed hydroxymethylation of phenol or benzamide with formaldehyde, because the acidic phenol and benzamide easily form their conjugate bases by addition of the basic catalyst in a preceding equilibrium step.     

Development of a particle agglutination assay system for detecting Japanese encephalitis virus-specific human IgM,using hydroxyapatite-coated nylon beads

Japanese encephalitis virus-specific IgM is a reliable indicator for serodiagnosis of Japanese encephalitis. A particle agglutination (PA) assay system was developed to detect anti-Japanese encephalitis virus IgM in human serum samples. The newly developed PA assay consisted of hydroxyapatite-coated nylon beads and V-bottom 96-well microplates. Hydroxyapatite-coated nylon beads were coated with Japanese encephalitis virus antigens. Japanese encephalitis virus antigen-coated, hydroxyapatite-coated nylon beads agglutinated in the IgM-captured wells when anti-Japanese encephalitis virus IgM-positive serum samples were used. A button pattern was formed at the bottom of the wells when anti-Japanese encephalitis virus IgM-negative serum samples were used. Thirty anti-Japanese encephalitis virus IgM-positive serum samples from Japanese encephalitis-confirmed cases were tested by the PA assay. All these serum samples were determined to be Japanese encephalitis virus IgM-positive. IgM titers determined by the PA assay corresponded to those determined by enzyme-linked immunosorbent assay. The titers were consistent in two independent PA assays. These results indicate that the newly developed PA assay is a reliable method for detecting anti-Japanese encephalitis virus IgM in human serum samples and that this assay will be a suitable diagnostic system especially in rural areas of Asia.     

Mevalonic acidemia: First case of Japan

Summary Mevalonic acidemia is a rare metabolic disorder due to mevalonate kinase deficiency which affects the biosynthesis of cholesterol and nonsterol isoprenes. We report the first case of Japan. The clinical course is characterized by intrauterine growth retardation, postnatal growth failure, intractable diarrhea, liver dysfunctions and death at three months of age. Dysmorphic features including triangular face, protrusion of forehead, hypertelorism, low set ears and micrognathism were noted. High mevalonic acid level was found by GC/MS.     

Cytokines and lipopolysaccharide induce nitric oxide synthase in cultured rat pulmonary artery smooth muscle.

In the current study, we describe cytokine and Escherichia coli lipopolysaccharide (LPS) induction of nitric oxide (NO) synthase mRNA levels in cultured smooth muscle from rat pulmonary artery (RPASM). Exposure of RPASM to interleukin-1 beta, interferon-gamma, or LPS alone did not significantly affect NO synthesis, as determined by nitrite concentrations in media. Exposure to tumor necrosis factor-alpha caused a modest (2x) increase in nitrite production. In contrast, exposure to a combination of the above three cytokines and LPS caused a large increase in NO synthesis. Exposure of RPASM to this combination caused an increase in mRNA levels of NO synthase (as described by Northern blot analysis with 32P-cDNA probe to an inducible form of NO synthase present in murine macrophages) that was apparent as early as 4 h. Expression of the induced gene product after exposure to the cytokine and LPS mixture was evident by significant increases in nitrite production at 12 h. Production of nitrite was completely abolished in the presence of NG-monomethyl-L-arginine (NMA), and this inhibition was reversible by the addition of excess L-arginine. NO synthase mRNA levels were not affected by NMA. The nitrite production induced by the combination of cytokines and LPS was abolished by pretreating cells with cycloheximide. These data indicate that a combination of cytokines and LPS affect expression of the gene for the inducible form of NO synthase in cultured RPASM.     

Membrane orientation of the precursor 74-kDa form of L-histidine decarboxylase

Objective We previously demonstrated that, when expressed in COS-7 cells, L-histidine decarboxylase (HDC), which has neither an amino terminal signal sequence nor a hydrophobic membrane anchor, was localized in the endoplasmic reticulum (ER), although its orientation in the membrane remains to be clarified. Methods & Results Protease digestion and immunofluorescence analyses of the cells, of which plasma membrane was selectively permeabilized, revealed that the amino terminal 50-kDa portion of HDC is hardly accessible to proteases and antibodies added exogenously from the cytosolic side. Green fluorescent protein fused with the carboxyl terminal 20-kDa region of HDC at its carboxyl terminus exhibited the same characteristics as native HDC. Conclusion These results indicate that HDC is tightly associated with the ER membrane with its carboxyl terminal region exposed on the cytosolic side. Received 22 November 2005; returned for revision 28 December 2005; accepted by A. Falus 22 January 2006     

Laboratory-scale mass production of a multi-micropatterned grafted surface with different polymer regions

In this article, we demonstrate laboratory-scale mass production of a regionally precise multi-micropatterned surface photo-graft-copolymerized with three water-soluble monomers based on the photochemistry of an iniferter, which means that it acts as an initiator, a transfer agent and a terminator, benzyl N, N-diethyldithiocarbamate. The surface was semi-automatically prepared using a combination of a custom-designed irradiation apparatus installed with a motor-controlled stage for a substrate and three photomasks with different line-patterned slits (number of slits 20, width 500 microm, length 10 mm), and carbon dioxide laser cutting apparatus. A particular region of poly(styrene-co-vinylbenzyl N,N-diethyldithiocarbamate) coated on a PET film was irradiated in a particular aqueous monomer solution while moving the irradiated portion stepwise after irradiation through each line of the photomask. Photo-graft-copolymerization was carried out sequentially with acrylic acid sodium salt (AANa), N-[3-(dimethylamino)propyl]acrylamide methiodide (DMAPAAm), and acrylamide (AAm) using differently patterned photomasks. Characterization of surface elemental distribution by X-ray photoelectron spectroscopy (XPS), and light microscopic visualization by dye staining revealed a microprocessed surface with 20 sets of micropatterns, each of which had three line regions grafted with three different polymers. The irradiation of a carbon dioxide laser manipulated via computer-aided design (CAD) software onto the microprocessed surface resulted in automatic circular cutting for each set of micropatterns to mass-produce multi-micropatterned substrates for the study of substrate-dependent endothelial cell responses.     

Effects of current intensity on behavioral and autonomic heat-loss responses in intracranial self-stimulating rats

We investigated the effects of current intensity on behavioral and autonomic heat-loss responses in intracranial self-stimulating (ICSS) rats. At an ambient temperature (Ta) of 22 degrees C, the tail vasodilation during ICSS behavior began at higher hypothalamic temperature (Thy) at higher current intensity. At a Ta of 36 degrees C, body extension, a typical heat-loss response in rats, appeared during ICSS behavior and frequently interrupted lever pressing. When the body extension first began, Thy was elevated if current intensity was high. In experiments in which current intensity was varied between two levels at a Ta of 22 degrees C, if current intensity was lowered after tail vasodilation began and Thy reached a steady level, the rat interrupted pressing the lever to lose heat through grooming or body extension. Rectal temperature (Tre) dropped in the process. If, on the other hand, current intensity was raised, the rate of lever pressing increased and Tre rose with tail vasoconstriction. The data suggest that the magnitude of the reward, as reflected in current intensity, affects both behavioral and autonomic heat-loss responses.     

Immunohistochemical study on tissue transglutaminase and copper-zinc superoxide dismutase in human myocardium: Its relevance to apoptosis detected by the nick end labelling method

The influence of free radicals on apoptosis was studied in the human heart; 45 autopsy cases were examined by the nick end labelling method (NELM) that detects DNA fragmentation. Immunostaining for copper-zinc superoxide dismutase (CuZn-SOD) and tissue transglutaminase (tTG) induced frequently during apoptosis were also studied. Positive immunoreaction for tTG was detected in mucinous degeneration of myocardial cells; these same cells were also positive for CuZn-SOD but negative for NELM. Myocardial cells showing basophilic alterations after haematoxylin and eosin staining were also positive for CuZn-SOD but negative for the other markers examined. Positive nuclear reaction by NELM was only observed in myocardial cells showing contraction band necrosis or irregularly shaped nuclei surrounding recent or long-standing infarcted foci. In these the other two markers were negative. Since mucinous degeneration lacks the distinguishing morphological features of apoptosis, immunoreactive tTG in this lesion may not imply that the cells are undergoing apoptosis. tTG can be induced in non-apoptotic conditions and may not be involved in apoptosis induced by infarction. Histological disassociation between CuZn-SOD expression and apoptosis suggests the possibility of a cytoprotective role played by endogenous CuZn-SOD against free radical generation in the human heart.