Biomedical subjects

C Morita

Publications and source records attributed to C Morita.

At least 19 recordsLinked to original sources

PubMed2003

A repertoire of pyridinium-phenyl-methyl cross-talk through a cascade of intramolecular electrostatic interactions.

Direct intramolecular cation-pi interaction between phenyl and pyridinium moieties in 1a(+) has been experimentally evidenced through pH-dependent (1)H NMR titration. The basicity of the pyridinyl group (pK(a) 2.9) in 1a can be measured both from the pH-dependent chemical shifts of the pyridinyl protons as well as from the protons of the neighboring phenyl and methyl groups as a result of electrostatic interaction between the phenyl and the pyridinium ion in 1a(+) at the ground state. The net result of this nearest neighbor electrostatic interaction is that the pyridinium moiety in 1a becomes more basic (pK(a) 2.92) compared to that in the standard 2a (pK(a) 2.56) as a consequence of edge-to-face cation (pyridinium)-pi (phenyl) interaction, giving a free energy of stabilization (DeltaDeltaG(o)pKa) of -2.1 kJ mol(-1). The fact that the pH-dependent downfield shifts of the phenyl and methyl protons give the pK(a) of the pyridine moiety of 1a also suggests that the nearest neighbor cation (pyridinium)-pi (phenyl) interaction also steers the CH (methyl)-pi (phenyl) interaction in tandem. This means that the whole pyridine-phenyl-methyl system in 1a(+) is electronically coupled at the ground state, cross-modulating the physicochemical property of the next neighbor by using the electrostatics as the engine, and the origin of this electrostatics is a far away point in the molecule-the pyridinyl-nitrogen. The relative chemical shift changes and the pK(a) differences show that the cation (pyridinium)-pi (phenyl) interaction is indeed more stable (DeltaDeltaG(o)pKa = -2.1 kJ mol(-1)) than that of the CH (methyl)-pi (phenyl) interaction (DeltaDeltaG(o)pKa = -0.8 kJ mol(-1)). Since the pK(a) of the pyridine moiety in 1a is also obtained through the pH-dependent shifts of both phenyl and methyl protons, it suggests that the net electrostatic mediated charge transfer from the phenyl to the pyridinium and its effect on the CH (methyl)-pi (phenyl) interaction corresponds to DeltaG(o)pKa of the pyridinium ion (approximately 17.5 kJ mol(-1)), which means that the aromatic characters of the phenyl and the pyridinium rings in 1a(+) have been cross-modulated owing to the edge-to-face interaction proportional to this DeltaG(o)pKa change.

Journal Article↗

PubMed2001

Postoperative magnetic resonance imaging of lumbar disc herniation: comparison of microendoscopic discectomy and Love's method.

STUDY DESIGN: We performed a study to compare the magnetic resonance imaging findings up to 24 weeks after microendoscopic discectomy or surgery using Love's method in patients with lumbar disc herniation. OBJECTIVES: The objective was to determine whether or not microendoscopic discectomy was minimally invasive with respect to the nerve roots, cauda equina, and paravertebral muscles by comparing the postoperative magnetic resonance imaging findings in patients treated by microendoscopic discectomy and the conventional Love's method. SUMMARY OF BACKGROUND DATA: We introduced microendoscopic discectomy as a minimally invasive surgical procedure for lumbar disc herniation in September 1998 and have obtained good results. Microendoscopic discectomy is superior to the conventional Love's method in that it reduces postoperative pain, shortens the duration of hospitalization, and allows earlier resumption of normal activities. However, the effect of microendoscopic discectomy on the nerves and paravertebral muscles has not been evaluated objectively. METHODS: Enhancement of the nerve roots and paravertebral muscles, as well as the configuration of the cauda equina at the level of herniation, was assessed on axial magnetic resonance images obtained with contrast enhancement using gadolinium-diethylenetriamine penta-acetic acid before surgery and 1, 4, 8, 12, and 24 weeks after surgery in 25 patients who underwent microendoscopic discectomy and 15 patients who were treated using Love's method. RESULTS: Increased enhancement of the nerve roots was seen in 50.0% of the microendoscopic discectomy group and 46.2% of the Love group at 1 week after surgery. Enhancement of the paravertebral muscles at the surgical site tended to persist for longer in the microendoscopic discectomy group than in the Love group. However, muscle enhancement was widespread in some patients from the Love group. Abnormalities of the cauda equina attributed to surgical invasion were seen in 12.5% of the microscopic discectomy group and 15.4% of the Love group at 1 week after surgery. CONCLUSIONS: Microendoscopic discectomy had an effect on the nerve roots and cauda equina that was comparable with that of Love's method. The magnetic resonance images of the route of entry failed to show that microendoscopic discectomy is appreciably less invasive with respect to the paravertebral muscles.

Adult↗

PubMed2001

Outside-to-inside signal through the membrane TNF-alpha induces E-selectin (CD62E) expression on activated human CD4+ T cells.

The membrane TNF-alpha is known to serve as a precursor of the soluble form of TNF-alpha. Although it has been reported the biological functions of the membrane TNF-alpha as a ligand, the outside-to-inside (reverse) signal transmitted through membrane TNF-alpha is poorly understood. Here we report a novel function mediated by outside-to-inside signal via membrane TNF-alpha into the cells expressing membrane TNF-alpha. Activation by anti-TNF-alpha Ab against membrane TNF-alpha on human T cell leukemia virus (HTLV) I-infected T cell line, MT-2, or PHA-activated normal human CD4(+) T cells resulted in the induction of an adhesion molecule, E-selectin (CD62E), on the cells with the peak of 12-24 h, which completely disappeared by 48 h. When wild-type or mutant membrane TNF-alpha (R78T/S79T) resistant to proteolytic cleavage was introduced into Jurkat or HeLa cells, E-selectin was induced by the treatment with anti-TNF-alpha Ab with the similar kinetics. Membrane TNF-alpha-expressing Jurkat cells also up-regulated E-selectin when brought into cell-to-cell contact with TNF receptor-expressing HeLa cells. Northern blot analysis and RT-PCR analysis showed that the membrane TNF-alpha-mediated E-selectin expression was up-regulated at the level of transcription. These results not only confirmed our previous findings of reverse signaling through membrane TNF-alpha, but also presented evidence that E-selectin was inducible in cell types different from endothelial cells. It is strongly suggested that membrane TNF-alpha is a novel proinflammatory cell surface molecule that transmits bipolar signals in local inflammation.

CD4-Positive T-Lymphocytes↗

PubMed2001

Targets of a protease inhibitor, KNI-272, in HIV-1-infected cells.

The targets of a protease inhibitor, KNI-272, in the HIV-1 life cycle were investigated in this study. Neither expression of HIV-1 Gag proteins nor production of virus particles was detected in cells infected acutely with HIV-1 cultured in the presence of KNI-272. Although HIV-1 proviral DNA was detected in the cells by PCR, the inhibitor depressed the amount of the proviral DNA in a concentration dependent manner. These results indicate that one of the targets of KNI-272 occurs in the stage before the expression of viral structural proteins. No direct inhibition of reverse transcription was found with the inhibitor. To confirm the inhibition of viral protease, persistently HIV-1-infected cells were cultured in the presence of the inhibitor and examined by electron microscopy for the morphology of HIV-1 particles. Doughnut-shaped immature particles were observed in the extracellular space of the cells, and disrupted semicircular shaped particles were also seen at the higher concentration of KNI-272. A bioassay for infectivity showed that the virus particles were not infectious, and immunofluorescent assay using anti-p17 antibody, that does not react with the precursor of Gag protein, revealed that Gag precursor p55 protein in the cells was not processed. Thus, KNI-272 blocked the maturation of viral particles. Consequently, KNI-272 has at least two inhibition targets in the stages of the HIV-1 life cycle.

Anti-HIV Agents↗

PubMed2001

Association of tumor necrosis factor receptor type II polymorphism 196R with Systemic lupus erythematosus in the Japanese: molecular and functional analysis.

OBJECTIVE: To investigate whether a polymorphism(s) or mutation(s) in the tumor necrosis factor receptor II (TNFRII) gene is involved in the pathogenesis of systemic lupus erythematosus (SLE). METHODS: All 10 exons of the TNFRII gene were analyzed by exon-specific polymerase chain reaction-single-strand conformation polymorphism, followed by nucleotide sequencing of exons that displayed aberrant bands. To analyze the function of the TNFRII polymorphisms, the full-length TNFRII complementary DNA of each allele was transfected in HeLa cells and then studied for specific binding of 125I-TNFalpha, as well as interleukin-6 (IL-6) production and cytotoxic activity after treatment with recombinant human TNFalpha. RESULTS: We identified 4 polymorphisms, at codons 56, 181, 196, and 232. The latter 2 had amino acid substitutions M196R and E232K, respectively. Only the 196R allele was significantly associated with SLE in our 105 Japanese SLE patients, with an allele frequency of 20.5%, compared with 12.6% in 99 healthy controls (P = 0.0335). More importantly, using TNFRII-transfected HeLa cells, we demonstrated significantly increased IL-6 production by 196R TNFRII compared with 196M TNFRII. The cytotoxic activity induced by 196R TNFRII was also increased compared with that of 196M TNFRII. This increase was achieved without affecting the binding affinity of TNFalpha to TNF-RII, as demonstrated by the finding that specific TNFalpha binding to the HeLa transfectants of 196R and 196M TNFRII was similar, with Kd values of 3.12 x 10(-10)M and 4.34 x 10(-10)M, respectively. CONCLUSION: These results suggest that 196R TNFRII, which transduces the signals of TNFalpha more effectively than does 196M TNFRII, is involved in the pathogenesis of SLE.

Adolescent↗

PubMed2001

Autoimmune hepatitis with membranous glomerulonephritis.

We report on a case of autoimmune hepatitis (AIH) associated with membranous glomerulonephritis. A 61-year-old woman was admitted because of peripheral edema, proteinuria and abnormal liver function test findings. A diagnosis of AIH was made on the basis of an elevation of aminotransferase and serum IgG levels, the presence of positive antinuclear antibody and the characteristic histological features of chronic active hepatitis. Histological examination of a renal biopsy specimen disclosed membranous glomerulonephritis with granular deposits of IgG, IgM, C3 and C1q along the capillary walls. This condition is rare in AIH and should be carefully distinguished from systemic lupus erythematosus.

Female↗

PubMed2001

An improved procedure of electron microscopic in situ hybridization for detecting adenovirus DNA.

Electron microscopic in situ hybridization (EM-ISH) is a useful method in determining the localization of a specific nucleic acid at the ultrastructural level. Since the EM-ISH protocol includes many steps, no standard protocol for EM-ISH is available yet. In this study, we optimized quantitatively the critical conditions with respect to embedding resin, nucleic acid labeling and hybridization reaction time, by using adenovirus-infected cells as the indicator cells. The optimal detection of an adenovirus-specific nucleic acid was obtained by overnight hybridization reaction on sections embedded in Lowicryl K4M resin. Random-primed-labeled probes improved the reactivity. At least 60% of virus particles in paracrystalline arrays was found to contain viral DNA. These arrays in adenovirus-infected cells are useful in evaluating quantitatively the efficiency of protocols of EM-ISH.

Adenoviridae↗

PubMed2001

Seroprevalence of antibodies against spotted fever group rickettsia among dogs and humans in Okinawa, Japan.

We evaluated serum antibodies against Rickettsia japonica in 517 dogs (430 stray dogs and 87 pet dogs) and 164 humans in Okinawa, Japan, by indirect immunofluorescence assay. The seropositive rate in stray dogs was significantly higher than that in pet dogs (30.7 versus 4.6%, P<0.01). This high prevalence rate is attributed to the understandably frequent environmental exposure of stray dogs to tick infestation. Human samples obtained from Okinawa and Sapporo also showed a significant difference in seropositive antibody percentages (45.1 and 12.0%, respectively, P<0.01). This result suggests that there has been pre-exposure to spotted fever group rickettsia in humans in Okinawa.

Animals↗

PubMed2001

First detection of Ehrlichia platys in dogs and ticks in Okinawa, Japan.

We investigated Ehrlichia platys infection of dogs and ticks in Okinawa, Japan. Using E. platys specific primers, E. platys and HE3-R, PCR-positive results were obtained with 32.0% (64/200) of blood samples of dogs and 3.8% (3/77) of ticks. The nucleotide sequences of the amplified DNA fragment from the dogs and the ticks infesting them were identical, and the sequence corresponded to that of the E. platys Gzh981 strain. We concluded that there is a cyclic maintenance of E. platys between dogs and ticks in Okinawa.

Animals↗

PubMed2001

Human babesiosis in Japan: epizootiologic survey of rodent reservoir and isolation of new type of Babesia microti-like parasite.

We have carried out epizootiologic surveys at various sites in Japan to investigate wild animals that serve as reservoirs for the agents of human babesiosis in the country. Small mammals comprising six species, Apodemus speciosus, Apodemus argenteus, Clethrionomys rufocanus, Eothenomys smithii, Crocidura dsinezumi, and Sorex unguiculatus, were trapped at various places, including Hokkaido, Chiba, Shiga, Hyogo, Shimane, and Tokushima Prefectures. Animals harboring Babesia microti-like parasites were detected in all six prefectures. Inoculation of their blood samples into hamsters gave rise to a total of 20 parasite isolates; 19 were from A. speciosus, and the other 1 was from C. rufocanus. Sequencing of the parasite small-subunit rRNA gene (rDNA) sequence revealed that 2 of the 20 isolates were classified as Kobe type because their rDNAs were identical to that of the Kobe strain (the strain from the Japanese index case). The other 18 isolates were classified as a new type, designated the Hobetsu type, because they all shared an identical rDNA sequence which differed significantly from both that of Kobe-type isolates and that of northeastern United States B. microti (U.S. type). The parasites with Kobe-, Hobetsu- and U.S.-type rDNAs were phylogenetically closely related to each other but clearly different from each other antigenically. The isolates from rodents were demonstrated to be infective for human erythrocytes by inoculation into SCID mice whose erythrocytes had been replaced with human erythrocytes. The results suggest that a new type of B. microti-like parasite, namely, the Hobetsu type, is the major one which is prevalent among Japanese wild rodents, that A. speciosus serves as a major reservoir for both Kobe- and Hobetsu-type B. microti-like parasites, and that C. rufocanus may also be an additional reservoir on Hokkaido Island.

Animals↗

PubMed2001

Characteristic features of the acetabular labrum in healthy children.

Radial magnetic resonance images of the acetabular labrum were obtained on 40 hips of healthy children. There were no right-left or male-female differences. In children aged 11 years or younger, the labrum on the antero-superior weightbearing portion was triangular in shape, and there was an insular-shaped or linear high-intensity area inside; on the mid-superior portion, the labrum appeared as a regular triangular, low-intensity area; and on the postero-superior portion, it was flat. In children aged 12 and 13 years, the shape of the labrum in each portion was similar to that of the younger children, but the high signal intensity area on the antero-superior portion appeared less frequently. The size of the labrum relative to the femoral head was greater in younger children.

Acetabulum↗

PubMed2000

CD56 directly interacts in the process of NCAM-positive target cell-killing by NK cells.

The role of CD56 in the process of target cell killing by NK cells has been investigated. Addition of NK cells to HuH28 cells, a CD56-expressing cell line, led to inhibition of the growth of the target cells, which exhibited morphological features of apoptosis. These changes were prevented by the addition of a polyclonal anti-NCAM to the cultures. Since neither Fas antigen expression nor apoptotic changes were induced by addition to a mixed culture supernatant of NK and target cells, both the Fas-Fas ligand system and soluble factors do not seem to participate in apoptosis in these circumstances. Increased secretion of interferon-gamma and tumour necrosis factor-alpha by NK cells must therefore have been suppressed by the presence of the polyclonal antibody. These results lead us to conclude that CD56, through homophilic binding, plays an important role in the process of target cell killing by an apoptosis mechanism.

Apoptosis↗

PubMed2000

Disinfection potential of electrolyzed solutions containing sodium chloride at low concentrations.

Electrolyzed products of sodium chloride solution were examined for their disinfection potential against hepatitis B virus (HBV) and human immunodeficiency virus (HIV) in vitro. Electrolysis of 0.05% NaCl in tap water was carried out for 45 min at room temperature using a 3 A electric current in separate wells installed with positive and negative electrodes. The electrolyzed products were obtained from the positive well. The oxidation reduction potential (ORP), pH and free chlorine content of the product were 1053 mV, pH 2.34 and 4.20 ppm, respectively. The products modified the antigenicity of the surface protein of HBV as well as the infectivity of HIV in time- and concentration-dependent manner. Although the inactivating potential was decreased by the addition of contaminating protein, recycling of the product or continuous addition of fresh product may restore the complete disinfection against bloodborne pathogens.

Acids↗

PubMed2000

Contrast-enhanced immunoelectron microscopy for Helicobacter pylori.

Since a method of contrast enhancement for immunoelectron microscopy has not been available in bacteriology, the morphological localization of proteins of Helicobacter pylori is not well known. In this report, we established a method of contrast enhancement in immunoelectron microscopy in this organism. Immunostained ultrathin sections are stained with a mixture of alcian blue and osmium tetroxide prior to staining with uranyl acetate. This method of staining provided good contrast enhancement of the bacterial cell wall and membrane without any loss of immunolabeled gold particles on the ultrathin section.

Acrylic Resins↗

PubMed2000

Detection of serum antibodies to Bartonella henselae and Coxiella burnetii from Japanese children and pregnant women.

The participation of Bartonella henselae and Coxiella burnetii in the pathogenesis of fever of unknown origin (FUO) and lymphadenopathy has not been completely clarified. Prevalence of these two agents in Japanese children is also unknown. Serum IgG and IgM antibodies to B. henselae and to C. burnetii were examined by the indirect fluorescence antibody assay. Enzyme immunoassay kits were used to detect serum IgG and IgA antibodies against Chlamydia trachomatis. Out of 200 healthy normal pregnant women, two (1.0%) had serum IgG antibodies to B. henselae, four (2.0%) to C. burnetii and 49 (24.5%) to C. trachomatis. Out of 29 patients with FUO, one (3.4%) had serum IgG antibodies to B. henselae, four (13.8%) to C. burnetii and none to C. trachomatis. Out of 31 patients with cervical lymphadenopathy, three (9.6%) had serum IgG antibodies to B. henselae, two (6.5%) to C. burnetii and none to C. trachomatis. Out of 22 patients with generalized lymphadenopathy, one (4.5%) had serum IgG antibodies to B. henselae, three (13.6%) to C. burnetii and none to C. trachomatis. Prevalences of serum antibodies to C. burnetii in the patients with FUO and generalized lymphadenopathy and to B. henselae in the patients with cervical lymphadenopathy were significantly higher than those of normal pregnant women (Welch's t-test; P<0.01). These two agents may have some roles in the pathogenesis of FUO and lymphadenopathy in Japanese children.

Adolescent↗

PubMed2000

Mannose binding lectin (MBL) gene mutation is not a risk factor for systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) in Japanese.

Mannose binding lectin (MBL) deficiency may be associated with increased susceptibility to infection and autoimmune disorders, such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). In the present study, we performed for the first systematic search for mutations in all the four exons of the MBL gene using polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) analysis. Of 49 healthy Japanese individuals studied, only the previously reported mutation at the codon 54 (substitution from Gly to Asp; G54D) was identified. The allele frequencies of G54D in 105 healthy Japanese individuals, 95 SLE patients and 59 RA patients, were 0.233, 0.226 and 0.178, respectively, which were not significantly different. In addition, two polymorphisms at positions of -550 and -221 in the promoter region were not associated with SLE and RA. It is unlikely that MBL deficiency plays a major role in the pathogenesis of SLE and RA in Japanese.

Alleles↗

PubMed2000

Development of battery charging system for the FE gun system of HVEM.

We have developed a reliable and efficient battery-charging system for the field emission (FE) gun system of high voltage electron microscopes, where the operating condition of the whole FE gun system can be controlled and monitored through a bi-directional optical fiber system, and the control and monitoring circuits located on the high potential of 1 MV are driven by rechargeable Ni-Cd batteries. The power transmission from ground to the high potential is performed through filter condenser circuit. Under the condition that the filter condenser current is limited to 0.2 A(rms) (root-mean-square value), it is possible to transmit the maximum power as high as 65 W, which is enough for the daily operation of the FE gun system. The charging circuit has a function of protecting the batteries from over-charging.

Journal Article↗

PubMed2000

Prevalence of spotted fever rickettsial antibodies in dogs and rodents in the Philippines.

Antibodies against spotted fever group rickettsiae have been detected in blood samples of dogs and rodents obtained from selected areas in the Philippines. In this serosurvey, the positive percentage rates are 8.3% (11/132) in dogs and 12.2% (6/49) in rats. Positive results were read from samples tested with Rickettsia japonica antigen. No positive result was obtained in blood samples of rats and house mice using R. akari antigen. The findings of this study are the first to confirm the detection of spotted fever group rickettsial antibodies in the Philippines.

Animals↗