Biomedical subjects
Publications and source records attributed to M Ohta.
The efficacy of chicken egg yolk immunoglobulins (yIg) from hens immunized with bovine rotavirus (BRV) serotype G6 (strain Shimane) or serotype G10 (strain KK-3) for protection against homologous BRV in calves was investigated. A significant protection by anti-BRV yIg having 6400 neutralizing antibody titer per dose could be achieved in calves (P < 0.01).
We examined the preventive effect of metoclopramide on the development of esophageal varices in a rat model. Thirty rats were divided into three groups: metoclopramide (7.5 mg/kg twice a day, intraperitoneally), control group I (saline 2 ml/kg twice a day, intraperitoneally), and control group II (incised lower esophageal sphincter and metoclopramide 7.5 mg/kg twice a day, intraperitoneally). On the 14th postoperative day, lower esophageal sphincter pressure in the metoclopramide group (8.6 +/- 1.4 cm H2O) increased more than in the control groups (5.4 +/- 0.5, 5.0 +/- 0.5 cm H2O, P < 0.01). Development of small collateral vessels from the spleen to the retroperitoneum was evident only in the metoclopramide group, as seen on the portography (P < 0.01). Histologically, the variceal area of the horizontal cross section of the esophagus in the metoclopramide group (0.62 +/- 0.26 mm2) was significantly smaller than in the controls (2.67 +/- 0.95, 2.78 +/- 0.82 mm2), determined using an image processor-analyzer for photographing histological specimens (P < 0.01). We also investigated the effect of metoclopramide on smooth muscle cells in the rat portal vein, using isometric-tension recording. Metoclopramide relaxed the smooth muscle precontracted with norepinephrine, in a concentration-dependent manner. Thus, metoclopramide inhibits the development of esophageal varices in this rat model due to both an increase in resistance of the lower esophagus and to development of small collaterals.
Morphological alterations in the pattern of liver cell gap junctions were examined in phalloidin-treated rats to assess the role of gap junctions in experimental intrahepatic cholestasis. Double-labelled fluorescent staining of gap junctions and F-actin were performed using a monoclonal antibody against rat hepatocyte connexin 32 and rhodamine-phalloidin. Immunoelectron microscopy, using the anti-connexin 32 antibody, freeze-fracture replica electron microscopy, and conventional electron microscopy were also performed. In phalloidin-treated rat livers, the specific immunofluorescent staining of connexin 32 was markedly decreased in the pericentral area after 1 day of phalloidin treatment and, after 5 days of phalloidin treatment, there was a decrease in connexin 32 staining in the entire hepatic lobule. On the other hand, F-actin staining at the cell periphery and at the bile canaliculi was markedly increased in the pericentral area of the hepatic lobule after 1 day of phalloidin treatment and in the entire lobule after 5 days of treatment. Immunoelectron microscopy showed that both sides of the cytoplasmic domains of gap junctions were stained with anti-connexin 32 antibody in controls, whereas, in cholestatic rats, only one side of the cytoplasmic domain of some gap junctions was stained with anti-connexin 32 antibody after 1 or 3 days of phalloidin treatment. No gap junctions were observed after 5 days of phalloidin treatment either by freeze-fracture replica electron microscopy or by conventional electron microscopy. These results indicate that with phalloidin treatment, hepatocyte gap junctions decrease, first in the pericentral area, and finally throughout the entire lobule.
We report herein the unusual case of a 45-year-old Japanese man whose chest X-rays revealed an abnormal shadow, increasing in size. A chest wall tumor was suspected, based on the findings of computed tomography (CT) of the thorax, the CT number of which was -137, chest roentgenogram, and an echograph. An open biopsy was performed to establish the final diagnosis. The resected tumor was a pedunculated pleural mass, yellowish in color, the pathological diagnosis of which confirmed a lipoma. Intrathoracic lipomas are rare, but pleural lipomas are seen even less frequently. CT, echography, and percutaneous needle biopsy have been found useful for diagnosing intrathoracic lipomas, but these examinations are not always adequate for confirming the final diagnosis. Consequently, tumor resection is essential for obtaining a pathological diagnosis.
A technique for laparoscopic splenectomy is described. The patient is placed in the right semidecubitus position and pneumoperitoneum is prepared. The splenic artery and vein are exposed near the hilum, using a laparoscopic ultrasonic dissector. The larger vessels are doubly ligated, and the spleen is resected and maneuvered into a nylon surgical sack; the sack is removed through a 2-cm incision along the midaxillary line. This procedure has been used for four patients requiring splenectomy for benign disease, and the outcome has been entirely satisfactory for all concerned.
1. High-Km aldose reductase purified from dog kidney inner medulla was easily converted into aldose reductase by incubation in the neutral buffer solution. 2. High-Km aldose reductase was found to be in multiple forms, and was separated into three kinds of species designated as a-, b- and c-forms by HPLC. 3. The a-form observed as a single peak by HPLC was assumed to be present in three forms (a1-, a2- and a3-forms), one was aldose reductase (a1-form) and the others were the precursors of aldose reductase (a2- and a3-form). 4. The b-form was rapidly converted into the a3-form, followed slowly by the a2-form and finally into the a1-form. 5. The c-form was either directly converted into the a1-form, or indirectly into the a2-form followed by the a1-form. 6. Four kinds of species (a2-, a3-, b- and c-forms) of high-Km aldose reductase were finally converted into aldose reductase (a1-form).
OBJECTIVES: To determine the long-term outcomes of patients with epithelial ovarian cancer. METHODS: A group of 298 patients with epithelial ovarian cancer were treated with combination chemotherapy between July 1979 and January 1986 at the Tokai Ovarian Tumor Study Group. Long-term results of these patients were analyzed. RESULTS: Five-year survival and 10-year survival rates of all the patients were 53% and 47%, respectively. Five-year survival and 10-year survival rates for stage I were 89% and 79%, 59% and 56% for stage II, 27% and 22% for stage III, and 9% and 9% for stage IV, respectively. Survival of mucinous and endometrioid cell types were decreased after 5 or more years. In stages II-IV, cisplatin-based chemotherapy produced better results than mitomycin-C, 5-Fu, cytarabine (MFC) therapy. CONCLUSIONS: Long-term results of epithelial ovarian cancer were not favorable even in mucinous and endometrioid cell types. For long-term results, cisplatin-based chemotherapy was effective in advanced ovarian cancer. Long-term follow-up of ovarian cancer is important and necessary.
Our previous studies have shown that Tm values for tauroursodeoxycholate (TUDC) and tauro beta-muricholate (T beta-MC) are more than two-fold higher than that for taurocholate (TC) in the rat. The present study attempted to clarify whether tauro alpha-muricholate (T alpha-MC) also has such an unusually large Tm value in the rat. Under nembutal anesthesia, male Wistar derived rats (body weight 280-300 g, 13 wks in age) were continuously infused with T alpha-MC solution. The infusion rate was raised stepwise every 20 min, until the bile flow began to decline. Bile was collected every 10 min and bile salt excretion rate was determined. The average of the highest three excretion values was assumed to be the Tm in each animal. The Tm value of T alpha-MC was found to be 2.86 +/- 0.36 mumol/min/100 g (mean +/- SD, n = 4), which was even greater than Tm values for TUDC (2.59 +/- 0.39 mumol/min/100 g, n = 4) and T beta-MC (1.93 +/- 0.31 mumol/min/100 g, n = 4) as we reported previously. The relationship between the bile flow rate (microliter/min/100 g, Y axis) and bile salt excretion rate (mumol/min/100 g, X axis) was highly linear [Y = (6.00 +/- 0.29) x +(6.60 +/- 1.88), P < 0.001, r = 0.95, n = 54]. The slope value for T alpha-MC (6.00 +/- 0.29 microliters/mumol) was significantly higher than that for TUDC (4.76 +/- 0.71 microliters/mumol) and was comparable to that for T beta-MC as we previously found for these bile salts in this rat strain. The results suggest that T alpha-MC has a very efficient transport system in this species as was observed for the other two bile salts that have a 7 beta-hydroxy group (TUDC and T beta-MC). This efficient transport system thus appears to be shared not only by bile salts specifically having a 7 beta-hydroxy group, but also by other bile salts such as T alpha-MC that have a 6 beta-hydroxy group but not a 7 beta-hydroxy group.
Male C57BL/6 mice were fed with normal diet (ND) or diets containing 0.3 or 0.5% ursodeoxycholic acid (UDCA) for 3 weeks. Glutathione S-transferase (GST) activities in the liver cytosolic fraction of these animals toward 1,2-dichloro,4-nitrobenzene (DCNB) as well as to 1-chloro,2,4-dinitrobenzene (CDNB) were significantly increased in a dose dependent manner in UDCA-treated groups compared with the control (ND-fed) animal group (one-way ANOVA). Reduced glutathione (GSH) levels tended to slightly decrease with UDCA diets but the difference did not attain a statistical significance (P > 0.05, one-way ANOVA). Twenty four hr survival rates after an oral challenge of 3.5 mg/kg of DCNB were significantly higher (P < 0.05, Chi-square test) in the two UDCA fed groups (10/10 for 0.5% group, 8/11 for 0.3% group) compared with the control group (3/11). Thus, UDCA appears to reduce the systemic toxicity of DCNB which is detoxified by the hepatic GST system. Although UDCA has been shown to exert hepatoprotective effects in experimental animals and humans in the past, to the best of our knowledge, the present study is the first report that UDCA reduces the systemic toxicity of a toxicant which is detoxified by the hepatic GST system. Although a direct proof is not available, it is most likely that the reduction of the systemic toxicity of DCNB was achieved by the increase in GST activity caused by UDCA feeding. This finding may open a new research field with regard to the unique biological properties of this bile salt in modulating hepatic detoxifying enzymes.
The fibroblast growth factor (FGF) receptor family consists of four members, FGFR-1, FGFR-2, FGFR-3 and FGFR-4, that are closely related receptor tyrosine kinases. We examined the expression of rat FGFR-4 mRNA in the brain by in situ hybridization and compared it with that of the mRNAs for other FGF receptors. In contrast with FGFR-1, FGFR-2 and FGFR-3 mRNAs which are expressed widely in the brain, the FGFR-4 mRNA in the brain is expressed preferentially in the medial habenular nucleus neurons. The present finding indicates that FGFR-4 has a function specific to the medial habenular nucleus.
Vertebrate lens tissues contain several species of acidic and neutral glycosphingolipids in relatively high amounts. However, the epithelia with capsule from dog and rhesus monkey lenses had a simpler composition and lower content of glycosphingolipids than whole lenses. Gangliosides and neutral glycosphingolipids in monolayer cultures of lens epithelial cells were also different from those in whole lenses. Although alpha-galactosyl (Gal alpha 1-3Gal-R) or Lewis(x) (Gal beta 1-4[Fuc alpha 1-3]GlcNAc-R) epitopes were found in glycosphingolipids from whole lenses, they were not detected in those from monolayer cultures of dog and rhesus monkey lens cells. In addition, significant changes in ganglio-series gangliosides were induced in monolayer cultures of both cells, where GM3 and GD3 were predominant. Immunofluorescence study revealed a characteristic distribution of cell surface gangliosides in confluent monolayers. These findings suggest that glycosphingolipid synthesis in lens epithelia is intrinsically different from that in cortical and nuclear fibres, and that the expression of Lewis(x) and alpha-galactosyl epitopes in glycosphingolipids appears to be associated with the differentiation of epithelial cells to fibres.
Subcloning, transposon insertion, and deletion analysis revealed that the Escherichia coli O9 rfb region is about 12 kb in size. The region encodes at least seven polypeptides of 89, 74, 55, 50, 44, 41 and 39.5 kDa. Southern hybridization analysis of rfb regions of E. coli O8 and O9, and Klebsiella O3 and O5 serotypes (all of these O polysaccharides are mannose homopolymers and the structures of the repeating unit of E. coli O9 and Klebsiella O3 are identical) showed that a central region specific for E. coli O9 and Klebsiella O3 is flanked by two regions common to all four. Complementation experiments using strains with known defects and specific tests for the enzymic activity showed that the 50 and 55 kDa polypeptides, encoded by the common region, are phosphomannomutase (PMM) and GDP-mannose pyrophosphorylase (GMP), respectively. Nucleotide sequencing of the region revealed the presence of two genes, rfbK and rfbM, analogous to the corresponding genes of Salmonella typhimurium. In E. coli O9, rfbK and rfbM encode proteins of 460 amino acids (50,809 Da) and 471 amino acids (52,789 Da). The amino acid sequence of GMP was conserved in RfbMs of E. coli O7 and Salmonella groups B, C1 and C2, CpsB of S. typhimurium, AlgA of Pseudomonas aeruginosa, and XanB of Xanthomonas campestris. The phylogenetic trees of PMM and GMP were different in topology and in the evolutionary distances from ancestors.
Lidocaine hydrochloride (LH), a local anesthetic, and acetylsalicylate (AcSAL), show antibacterial activity for both gram-negative and gram-positive bacteria. Kinetic studies indicated that antibacterial activity of LH was different from that of AcSAL. A subinhibitory concentration of LH and AcSAL enhanced the sensitivity of Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa to novobiocin and nalidixic acid. The synergistic effect of AcSAL with novobiocin and nalidixic acid was higher than that of LH. The effect of both drugs on the membrane potential of inner membrane was also studied using inverted membrane vesicles of bacteria. Both LH and AcSAL depolarized the membrane potential after the vesicles were energized with nicotinamide adenine dinucleotide. However, unlike AcSAL, pre-treatment of vesicles with LH had no effect on the generation of membrane potential. These results suggest that depolarization of the cytoplasmic membrane, preceded by the permeabilization of the outer membrane for gram-negative bacteria, is associated with antibacterial activity of LH and AcSAL. The difference in actions of LH and AcSAL was discussed.
Various chemotypes (Re, Rd2, Rd1P-, Rd1, RcP-, Rc, Rb3, Rb2, Rb1, and Ra) of R-form lipopolysaccharides (LPSs) of Salmonella spp. were crystallized by treatment with 70% ethanol containing 250 mM MgCl2, and crystals of the LPSs were observed electron microscopically and analyzed by electron diffraction and synchrotron X-ray diffraction. All the LPSs tested formed three-dimensional crystals showing very similar shapes; hexagonal plate, solid column, discoid, square or rectangular plate, lozenge plate and truncated hexangular or rectangular pyramid forms. Electron diffraction patterns from the hexagonal plate crystals of all these LPSs obtained by electron irradiation from the direction perpendicular to the basal plane showed that they consist of hexagonal lattices with the lattice constant of 4.62 A. The crystals of all the LPSs thus formed gave ring-like X-ray diffraction patterns because of their small sizes. The long-axis values were calculated from the X-ray diffraction patterns from crystals of all the LPSs in the low-angle region and they corresponded roughly to the length of the proposed primary chemical structures of the R cores of the LPSs. The volume occupied by a single molecule of all the LPSs were calculated from the molecular weights based on the proposed structures and the crystallographic data obtained by electron diffraction, X-ray diffraction, and density determination.
We established six human T cell lines derived from rheumatoid arthritis synovial fluid (RASF). Phenotypically, T cell receptor (TCR) gamma delta T cells occupied the majority of these lines and most of them expressed the TCR V delta 1 molecule. In contrast, V delta 2+ T cells, the majority population of peripheral blood gamma delta T cells, were rarely detected in these lines. To study the immunobiological roles of RASF V delta 1+ T cells in RA development, their cytotoxic profile was studied. The results showed that these T cells selectively lysed Daudi, but not K562 cells. The cytotoxic response was MHC-unrestricted, and was inhibited by anti-CD3 MoAb. Moreover, the cold target inhibition assay showed that the cytotoxicity was competitively inhibited by autologous and allogeneic primarily cultured RA synovial cells as well as synovial sarcoma and chondrosarcoma lines. However, PBL did not inhibit this cytotoxicity. These data suggest that V delta 1+ T cells in RASF may recognize the antigen which is commonly expressed on the surface of Daudi and the cells derived from RA synovium. We can assume that the cytotoxic V delta 1+ T cells are selectively expanded in RASF, playing a significant role for the pathogenesis of certain RA cases.
Tachyplesin I, a polycationic antimicrobial peptide isolated from hemocytes of horseshoe crabs, kills bacteria by disrupting the membrane potential of the cytoplasmic membrane. The present study shows that, among 36 oral streptococcal strains, 12 of 21 Streptococcus sanguis, 3 Streptococcus mutans, 9 Streptococcus salivarius and 3 Streptococcus milleri strains were susceptible to tachyplesin I, whereas 9 S. sanguis strains were resistant. Interestingly, these resistant strains include the clinical isolates from both Kawasaki disease and Behçet patients. According to the time-kill study, tachyplesin I inhibited irreversibly the growth of S. sanguis, S. mutans and S. salivarius strains within 20 min and an S. milleri strain within 80 min. Although it has been suggested that Escherichia coli cultured in rich media were more susceptible to tachyplesin I, the present results show that only 3 S. milleri strains were more sensitized to tachyplesin I in a glucose-supplemented medium, and other tested strains were not. Similarly, only 4 strains were more resistant to tachyplesin I in saline than these were in a rich medium.
Auditory brainstem responses (ABRs) were examined in 30 schizophrenic patients and 29 normal subjects. The psychotic symptoms were assessed by the Brief Psychiatric Rating Scale (BPRS) and the Scale for Assessment of Negative Symptoms (SANS) in the patients. At least one of the waves I, II or III was found missing on either side at 80 dBHL (hearing level) in 8 (27%) of the patients but in only one (3%) of the normal subjects. There was a significant association between the missing peaks and the BPRS negative symptom cluster or the total score of the SANS. These results suggest that some schizophrenics, especially those with negative symptoms, have an abnormality of input processing of auditory information in the lower brainstem.
A clinical isolate of Serratia marcescens (TN9106) produced a metallo beta-lactamase (IMP-1) which conferred resistance to imipenem and broad-spectrum beta-lactams. The blaIMP gene providing imipenem resistance was cloned and expressed in Escherichia coli HB101. The IMP-1 was purified from E. coli HB101 that harbors pSMBNU24 carrying blaIMP, and its apparent molecular mass was calculated to be about 30 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Kinetic studies of IMP-1 against various beta-lactams revealed that this enzyme hydrolyzes not only various broad-spectrum beta-lactams but also carbapenems. However, aztreonam was relatively stable against IMP-1. Although clavulanate or cloxacillin failed to inhibit IMP-1, Hg2+, Fe2+, or Cu2+ blocked the enzyme's activity. Moreover, the presence of EDTA in the reaction buffer resulted in a decrease in the enzyme's activity. Carbapenem resistance was not transferred from S. marcescens TN9106 to E. coli CSH2 by conjugation. A hybridization study confirmed that blaIMP was encoded on the chromosome of S. marcescens TN9106. By nucleotide sequencing analysis, blaIMP was found to encode a protein of 246 amino acid residues and was shown to have considerable homology to the metallo beta-lactamase genes of Bacillus cereus, Bacteroides fragilis, and Aeromonas hydrophila. The G+C content of blaIMP was 39.4%. Four consensus amino acid residues, His-95, His-97, Cys-176, and His-215, which form putative zinc ligands, were conserved in the deduced amino acid sequence of IMP-1. By determination of the amino acid sequence at the N terminus of purified mature IMP-1, 18 amino acid residues were found to be processed from the N terminus of the premature enzyme as a signal peptide. These results clearly show that IMP-1 is an enterobacterial metallo beta-lactamase, of which the primary structure has been completely determined, that confers resistance to carbapenems and other broad-spectrum beta-lactams.