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Biomedical subjects

M Mochizuki

Publications and source records attributed to M Mochizuki.

At least 19 recordsLinked to original sources

Juvenile amyotrophy of the distal upper extremity: pathologic findings of the dura mater and surgical management.

STUDY DESIGN: Five cases of juvenile amyotrophy of the distal upper extremity were reviewed retrospectively to elucidate the pathophysiology of spinal cord dysfunction and the results of surgical management. OBJECTIVES: To clarify the pathogenesis of juvenile amyotrophy of the distal upper extremity and to present the results of a new surgical treatment. SUMMARY OF BACKGROUND DATA: Hirayama first reported this disorder in 1959. It is characterized by juvenile onset, slow progression, and involvement of the unilateral distal upper extremity. Recently, compression of the cervical spinal cord during neck flexion was implicated as a possible etiology of the disorder, but the exact etiology is still unknown. The value of surgical treatment for patients with juvenile amyotrophy of the distal upper extremity has not been established. METHODS: The clinical and radiographic characteristics of five patients with juvenile amyotrophy of the distal upper extremity were examined. All five patients were treated surgically with duraplasty in combination with posterior spinal fusion. Dynamic and computed tomographic myelography were performed before and after surgery. Intraoperative ultrasonography and conductive spinal cord evoked potentials were recorded before and after duraplasty. The surgical results and the histology of the resected dura were studied. RESULTS: Myelograms taken with the neck in a neutral position showed that the spinal cord was flattened in all five patients. When the neck was flexed, the dura and the spinal cord were compressed further. Intraoperative ultrasonography during neck flexion revealed an anterior shift of the spinal cord and decreased spinal cord pulsation. Amplitude of the conductive spinal cord evoked potentials decreased with neck flexion but increased after dural incision. Histologically, the dura appeared abnormal in that it contained few elastic fibers without the normal wavy structure. CONCLUSIONS: Juvenile amyotrophy of the distal upper extremity was characterized by inelastic dura that constricts and compresses the cervical spinal cord when the neck is in either a neutral or a flexed position. Abnormal dura appeared to be the cause of juvenile amyotrophy of the distal upper extremity. Duraplasty with spinal fusion are proposed as treatments.

Adolescent

Detection of mutagenicity in Ames test using a metalloporphyrin/oxidant model system for cytochrome P450.

A chemical model system for cytochrome P450, a porphyrin and an oxidant, was used in Ames assay as a substitute for S9 mix. In the presence of tetrakis(pentafluorophenyl)porphyrinatoiron(III) chloride [Fe(F5P)Cl] and tert-butyl hydroperoxide (t-BuOOH), mutagenicity of N-nitrosodibutylamine (NDB) in Salmonella typhimurium TA1535 was detected. The mutagenicity depended on the pre-incubation period, and also on the concentration of an oxidant and of bacteria. In the chemical model system, pH affected the mutagenicity of NDB, which suggested that as observed in an enzymatic activating system, the mutagenicity was due to the labile alkylating species which was derived from NDB activated in the chemical activation system and was sensitive to pH. Under the optimum conditions; a higher concentration of an oxidant, a higher concentration of bacterial culture, and a weakly acidic medium, mutagenicity of N-nitrosodipropylamine in S. typhimurium TA1535 was also detected. Besides N-nitrosodialkylamines, 2-aminofluorene (2-AF) and benzo[a]pyrene (BaP) were also used as mutagens. Mutagenicity of 2-AF and BaP in S. typhimurium TA1538 were both detected in the same system as used in detecting the mutagenicity of N-nitrosodialkylamines. Ames test using a metalloporphyrin/oxidant model system makes it possible to detect mutagenicity derived from both base pair substitution mutagens and frameshift mutagens without using enzymatic activating system. These results demonstrate that the assay with the chemical model system is useful in detecting unstable unknown active mutagens or investigating the mechanisms of the metabolic pathway of mutagens or carcinogens in a protein-free medium.

Benzo(a)pyrene

Immediate and chronic results of cutting balloon angioplasty: a matched comparison with conventional angioplasty.

BACKGROUND: At the initial stages of percutaneous transluminal coronary angioplasty (PTCA), several studies reported on the feasibility of coronary artery incision and dilatation leading to the extension of the PTCA technique. HYPOTHESIS: This study was designed to determine the immediate and chronic results of cutting balloon (CB) angioplasty. METHODS: This procedure was performed on 127 lesions in 110 patients (male 83%, age 61.8 +/- 9.3 years). RESULTS: The overall procedural success rates for the CB were 93.7% (119 lesions) and 92.7% (102 patients), while solitary CB without pre- and/or postdilatation was 76.4% (91 lesions). There was one major in-hospital complication (Q-wave myocardial infarction, 0.9%), but there were no deaths or emergency coronary artery bypass graftings. Significant angiographic dissections (> or = grade C) occurred in four patients, and coronary perforation occurred in one. The successfully treated CB group (95 lesions) was matched with the successful conventional angioplasty group (PTCA group) for chronic result assessment in regard to reference vessel size and lesion characteristics. In the CB group, postprocedural minimal luminal diameters were significantly larger and the percentage of stenosis at the stenotic site was significantly lower compared with the PTCA group. Restenosis occurred in 22 lesions (23.1%). This showed a significantly lower restenosis rate compared with the PTCA group (42.1%). In addition, the restenosis rate of the CB without inclusion of the pre- and/or postdilatation-treated lesions was 19.7%. CONCLUSIONS: (1) Cutting balloon angioplasty procedures can be performed with high success rates with few major inhospital events. (2) The restenosis rate in the CB group was significantly lower compared with the PTCA group.

Angioplasty, Balloon, Coronary

Determination of the PCO2-dependent component of the H+ concentration in venous and arterial blood plasma.

In normal venous blood plasma, the regression line of [H+] plotted against the PCO2 was linear against the square root of PCO2. Sequential measurements in venous and arterial blood of PCO2 and [H+] showed that the venous-arterial (V-A) difference in [H+] was linearly related to the V-A difference in the square root of PCO2, the regression line having the same slope as that of the venous [H+] plotted against the square root of PCO2. These findings suggested that the venous [H+] on the regression line represents the PCO2-dependent component, of [H+], [H+]*. The PCO2-independent component, delta [H+], can then be given by subtracting [H+]* from the measured [H+]. The delta [H+] in venous blood agreed well with that in arterial blood with a correlation coefficient of 0.99, supporting the validity of the value of [H+]*.

Adult

Characteristics of T cell lines established from skin lesions of Behçet's disease.

Hypersensitivity to a streptococcal antigen is postulated to be the pathogenesis of Behçet's disease. We analyzed T lymphocyte-phenotypes infiltrated in cutaneous pustular lesions in Behçet's disease and found that CD4+ T cells were predominant components although CD8+ T cells were also present in the lesion. In addition, we established T cell lines from pustular lesions of the four patients with a streptococcal antigen, KTH-1. Two of the cell lines showed the cell surface markers of CD8+TCR alpha beta +, and expressed mRNAs for interleukin (IL)-8, tumor necrosis factor (TNF) alpha, and perforin. Two other T cell lines expressed the cell surface markers for CD4+TCR alpha beta +. Cytokine expression pattern of the two CD4+ T cell lines revealed that one is Th1 type and the other is Th2 type. The Th2 type cell line showed marked proliferation with autologous peripheral blood mononuclear cells, suggesting that the self-reactive T cells play some role on the pathogenesis of Behçet's disease.

Adult

Non-restricted T cell receptor (TCR)-V alpha and -V beta gene usage in patients with pulmonary sarcoidosis.

Sarcoidosis is a systemic granulomatous disease of unknown etiology characterized by the pronounced accumulation of CD4+ T cells and macrophages in the affected organs. TCR variable (V) alpha and V beta gene usage in patients with sarcoidosis is still a matter of discussion. In this investigation, we analysed TCR-V alpha and -V beta gene usage in bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells (PBMC) of 30 patients with active pulmonary sarcoidosis using an adapter ligation method, reverse transcriptase-polymerase chain reaction (RT-PCR), and sequence-specific oligonucleotide probe (SSOP) analyses. There was no significant difference in TCR-V alpha or -V beta gene usage between BALF (n = 12) or PBMC (n = 27) of patients and PBMC of healthy subjects (n = 10). Neither selective TCR-V alpha nor -V beta expansion was observed in the paired BALF and PBMC from seven of nine patients. However, selective expansions were observed in a few TCR-V alpha or -V beta subsets in the BALF or PBMC of some individuals. Although a modest increase in a few TCR-V alpha or -V beta subsets was observed in the BALF or PBMC of some individuals, the increased TCR-V alpha or -V beta subsets were not closely associated with the HLA-DRB1, DQA1, DQB1, and DPB1 alleles of these patients. These results suggest that TCR-V alpha or -V beta gene usage is not restricted in both lung and peripheral blood in the majority of patients with active pulmonary sarcoidosis.

Adult

Human T-cell lymphotropic virus type 1 can infect primary rat retinal glial cells and induce gene expression of inflammatory cytokines.

PURPOSE: To examine whether or not retinal glial cells can be infected by human T-cell lymphotropic virus type 1 (HTLV-1) and test the possibility that HTLV-1-infected retinal glial cells are involved in the pathogenesis of HTLV-1 uveitis (HU). METHODS: We tested infection of HTLV-1 by a standard coculturing method using WKAH rat retinal glial cells and irradiated MT-2, a human T cell line that produces HTLV-1. Infection was confirmed by detecting the integrated HTLV-1 provirus, using polymerase chain reaction (PCR), viral gene expression, using reverse transcriptase-PCR (RT-PCR) and HTLV-1 p19 ELISA, and by identifying the HTLV-1-infected glial cells by immunofluorescence cytochemistry and in situ hybridization. Changes in cytokine gene expression were studied by RT-PCR. RESULTS: Using a semiquantitative PCR of HTLV-1 provirus sequence, we found that 2.6% of the retinal glial cells were infected at 3 days after infection, followed by a gradual decrease in the percentage with an extended period of culture up to 4 weeks. This time course of infection was also verified by RT-PCR and ELISA studies that detect viral mRNA expression and protein production, respectively. Expression of HTLV-1 gag protein and tax mRNA was detected in a part of glial cells by indirect immunofluorescence cytochemistry and in situ hybridization, respectively. RT-PCR analysis of cytokine gene expression revealed that gene expression of IL-6, CINC-1 (Gro, KC), and TNF-alpha were induced in these cells, with a peak at 3 weeks after infection. CONCLUSION: These results provided supportive evidence for the theory that the infection of retinal glial cells by HTLV-1 and subsequent production of inflammatory cytokines could be one contributing factor for the development of the unique clinical features of HU. A better understanding of the specific roles of the inflammatory cytokines in the pathogenesis of HU would be beneficial in the treatment and control of this disease.

Animals

Replication of feline syncytial virus in feline T-lymphoblastoid cells and induction of apoptosis in the cells.

Feline syncytial virus (FSV) was isolated from feline peripheral blood mononuclear cells of FSV-seropositive cats. When the susceptibility of feline T-lymphocytes to FSV was examined using three strains of FSV, FSV antigens were detected in the FSV-infected T-lymphoblastoid cells. Further, a diversity of biological properties, including replication kinetics and syncytia formation, was noted among the strains, and condensation of chromatin and the fragmentation of cellular DNA were observed in the infected cells. From these data, we conclude that FSV is lymphotropic and can induce apoptosis in the lymphocytes.

Animals

Cystic pulmonary metastases of endometrial stromal sarcoma of the uterus, mimicking lymphangiomyomatosis: a case report with immunohistochemistry of HMB45.

A case of endometrial stromal sarcoma (ESS) showed cystic pulmonary metastases mimicking lymphangiomyomatosis (LAM). A 58-year-old female, who had undergone total hysterectomy for low-grade ESS 16 years previously, had repeated bouts of pneumothorax. Multiple thin-walled cysts in the peripheral lung were revealed by radiological examinations. In an open-lung biopsy specimen, cystic lesions were surrounded by layers of spindle-shaped cells of varying thickness that resembled LAM. However, in addition to subtle histologic differences from LAM, HMB45 (antimelanoma antibody) showed positive in LAM (n = 3), but was negative in ESS (n = 2) and the cystic lesions of this case. Using myogenic markers (desmin and alpha-smooth muscle actin), metastatic ESS could be immunohistochemically differentiated from mesenchymal cystic hamartoma (n = 1). HMB45 immunohistochemistry is useful in the differential diagnosis of cystic pulmonary lesions.

Actins

Isolation from diarrheal and asymptomatic kittens of three rotavirus strains that belong to the AU-1 genogroup of human rotaviruses.

A survey of 143 stool specimens collected during a 17-month period in Japan from diarrheal and asymptomatic kittens identified three rotavirus strains that were similar in their genomic RNA constellation to human rotavirus AU-1 (G3P3[9]), lending strong support to the view that rotaviruses belonging to the same genogroup are circulating in both the human and the feline population.

Animals

Epitope mapping of a monoclonal antibody specific to feline panleukopenia virus and mink enteritis virus.

To obtain monoclonal antibodies (MAbs) specific to feline panleukopenia virus (FPLV) and mink enteritis virus (MEV), 15 hybridomas secreting MAbs against MEV-Abashiri were established and the properties of the MAbs were analyzed. The cross-reactivity of MAbs revealed that one MAb, P2-215 was specific for FPLV and MEV, whereas the remaining fourteen MAbs reacted with canine parvovirus (CPV), FPLV, and MEV. Epitope analyses using various CPV/MEV chimeric viruses revealed that the MAb P2-215 recognized the epitope comprised of amino acid 93-Lys in VP2, which is known to be FPLV and MEV-specific.

Amino Acid Sequence

Antigenic and plaque variations of serotype II feline infectious peritonitis coronaviruses.

Three feline coronavirus (FCoV) isolates KUK-H, M91-266, and M91-267 were examined to elucidate their biological and antigenic properties as well as disease potential in cats. Immune stainings of virus-infected cells by using FCoV type-specific monoclonal antibodies indicated that their antigenic specificity was serotype II. However, antigenic variations among these serotype II FCoVs were detected by neutralization assay with hyperimmune antisera against FCoVs and canine coronaviruses, and with experimentally infected cat sera; there were two subtypes in serotype II FCoVs. The isolates efficiently grew in fcwf-4 cell culture showing lytic CPE enough to form distinct plaques: when measured 48 hr after infection, plaque sizes of both M91-266 and M91-267 were approximately 1 mm in diameter, and a mixture of small (less than 1 mm in diameter) and large (approximately 3 mm in diameter) plaques were produced in the case of KUK-H. Strains KUK-H, M91-266 and M91-267 produced feline infectious peritonitis (FIP) in 50%, 67% and 89% of experimentally inoculated kittens, respectively. Furthermore, 80% of the kittens inoculated with the small plaque former of KUK-H developed FIP accompanied by more prominent clinical signs as well as pathological changes when compared with 28.6% of kittens inoculated with the large plaque former. These results suggest that serotype II FIPVs producing smaller size of plaques are more virulent than those producing larger size of plaques.

Animals

Identification of canine calicivirus capsid protein and its immunoreactivity in western blotting.

A canine calicivirus (CaCV) isolated in Japan, designated as CaCV No. 48 strain, was propagated in MDCK cells and purified by CsCl equilibrium gradient centrifugation. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the purified samples revealed the presence of only one major species of viral protein of about 60 kilodaltons after Coomassie staining. The same band, presumably that of the capsid protein, was detected by western blotting using a mouse hyperimmune serum. This capsid protein was synthesized in MDCK cells as early as 2 hr post-inoculation. Experimental infection of dogs resulted in the production of anti-CaCV antibodies which were detected by microneutralization test and western blotting. Likewise, serosurvey revealed not only the presence of neutralizing antibodies but also reactivity of the field sera against the capsid protein of the purified virus. These results indicate that the capsid protein of CaCV No. 48 strain is immunogenic and could be detected by antibodies in western blotting.

Animals

CD8+ T lymphocyte counts as an adjunctive predictor of cytomegalovirus retinitis in patients with acquired immunodeficiency syndrome.

We encountered a case of cytomegalovirus (CMV) retinitis with spontaneous regression in association with an increased number of CD8+ but not CD4+ T lymphocytes in a patient with acquired immunodeficiency syndrome (AIDS). Thus, we examined the number of CD4 and CD8 counts at the diagnosis of CMV retinitis and compared with those of Pneumocystis carinii pneumonia (PCP). All 21 patients with a confirmed diagnosis of PCP (22 episodes) and 14 CMV retinitis patients were included in this study. Although PCP occurred after depletion of CD4 count to below 200/microl, CD8 count at that time varied widely and remained normal in some patients (median: 276.5/microl, range: 133-900/microl). In contrast, as CMV retinitis occurred after further depletion of CD4 count to below 50/microl, CD8 count decreased to below 500/microl (median: 238/microl, range: 43-448/microl) (p<0.05; PCP vs CMV retinitis). The values of the sensitivity and specificity for CMV retinitis obtained at the cutoff point of either 400/microl in CD8 count and 50/microl in CD4 count were similar. When these two cutoff points were combined, the positive predictive value was 46%. Monitoring of CD8 count may prove valuable as an additional predictor of CMV retinitis in patients with AIDS after CD4 depletion.

AIDS-Related Opportunistic Infections

Immunotherapy for Behçet's disease.

Behçet's disease is an inflammatory disorder affecting many organs including the eye and one of the most common sight-threatening causes in countries around the Mediterranean basin and in the Asia including Japan, Korea and China. A number of clinical and laboratory findings suggest the significant involvement of the immune alterations in the pathophysiology and the pathogenic mechanisms of Behçet's disease. The immune alterations demonstrated in the disease include the alteration of the T cell circuitry and abnormal functions of the leukocyte. Because immunologic processes are believed to be in the chain of events in the manifestation of Behçet's disease, various agents capable of modulating the immune responses have been used to treat the disease. These drugs include corticosteroids, colchicine, cytotoxic agents, and immunophilin ligands (cyclosporine and FK506). This paper reviews the experimental and clinical investigations to analyze the immunopharmacological activities of these immunosuppressive agents in animal models and in patients with Behçet's disease.

Adrenal Cortex Hormones

Immunologic and virologic characterization of the primary infiltrating cells in the aqueous humor of human T-cell leukemia virus type-1 uveitis. Accumulation of the human T-cell leukemia virus type-1-infected cells and constitutive expression of viral and interleukin-6 messenger ribonucleic acids.

PURPOSE: To characterize immunologically and virologically the infiltrating cells in the aqueous humor of patients with human T-cell leukemia virus type-1 (HTLV-1) uveitis (HU). METHODS: With their informed consent, patients had 0.1 ml of the aqueous humor in the anterior chamber collected with a needle under an operating microscope. An aliquot of the collected sample from patients without steroid therapy was examined by May-Giemsa staining and immunocytochemically. The presence of the HTLV-1-infected cells was investigated by polymerase chain reaction (PCR) using the gag and pol regions of the provirus genome. The population of the infected cells was compared by PCR testing the amplification of the virus genome from 60 cells, or determining the endpoint of successful amplification of the twofold dilution series of the samples, collected from the aqueous humor and peripheral blood mononuclear cells (PBMCs), which were obtained at the same time. Expression of viral and cytokine genes was studied by reverse transcriptase-PCR (RT-PCR). The interleukin-6 (IL-6) level in the aqueous humor of patients with HU and control subjects was measured by a high-sensitivity enzyme-linked immunosorbent assay kit. RESULTS: The number of the infiltrating cells ranged from 475 to 3563 (mean = 2111) per 0.1 ml of aqueous humor, and all the identifiable cells were lymphocytes. Most of them were CD3-positive T cells (mean = 78%), whereas CD4-positive cells constituted less than half (mean = 35.3%). HTLV-1 provirus was detected by PCR in the infiltrating cells of 36 of 38 patients with HU tested, whereas it was detected in 1 of 4 seropositive patients with other entities of uveitis. A higher population of the infected cells in the aqueous humor than in the PBMC was found in seven of nine patients with HU by two independent approaches. Expression of HTLV-1 env or pX genes or both was shown in all 12 patients with HU tested by RT-PCR. IL-6 messenger ribonucleic acid (mRNA) was detected by RT-PCR in 10 of these 12 patients, whereas those of interleukin-1 alpha, interleukin-2, interleukin-4, and tumor-necrosis factor-alpha were not, and that of interferon-gamma was detected in only 1 patient. The IL-6 level was elevated significantly in the aqueous humor of nine patients with HU compared with that of five control subjects (520.2 +/- 841 pg/ml versus 2.77 +/- 1.59 pg/ml, P < 0.01 by Mann-Whitney test). CONCLUSIONS: HU is characterized by lymphocytic infiltration with a predominance of T cells and by the presence and probable accumulation of HTLV-1-infected lymphocytes in the affected eye. Production of viral antigens and IL-6 by the infiltrating cells could be responsible for the development of HU.

Adult

Use of topical FK506 in a corneal graft rejection model in Lewis rats.

PURPOSE: To evaluate the immunosuppressive effect of topical FK506 on allograft corneal rejection in rats. METHODS: Lewis rats were used as recipients and Fisher rats as corneal graft donors. In Experiment 1, all rats received intraperitoneally FK506 (0.3 mg/kg per day) for 7 days to ensure equal baseline parameters. The rats then were assigned randomly to treatment with topical 0.3% FK506 or vehicle alone. In another set of experiments, rats were treated only with topical treatment. The grafts were inspected by clinical evaluation. Corneas obtained at the time of maximum rejection were used for histology and immunohistochemistry. RESULTS: The selected combination of rat strains caused 100% graft rejection in untreated animals within 2 weeks after the penetrating keratoplasty. In the treated animals, rejection was delayed until the end of topical therapy. One third of corneal grafts remained clear until day 30. Histologic and immunohistochemical studies confirmed the clinical evaluations. Untreated rat corneas had a large number of infiltrating helper-inducer T cells, macrophages, interleukin-2 receptor-expressing cells, and Ia-antigen-expressing cells. At the same timepoint, topically treated corneas showed a limited inflammatory response characterized by a 2/3 reduction in the number of infiltrating helper and cytotoxic cells, and a five-fold decrease in the expression of class I and class II major histocompatibility antigens. CONCLUSIONS: Topical FK506 treatment is an effective way of preventing corneal graft rejection in the Lewis rat corneal graft model. It shows promise as a drug to prevent corneal graft rejection in humans.

Administration, Topical

[Human T-lymphotropic virus type 1 uveitis in children].

We report here five pediatric patients with human T-lymphotropic virus type 1 (HTLV-I) uveitis. The patients were one boy and four girls aged between 3 and 14 years. The transmission route was considered to be breast feeding from their mothers. All patients had unilateral uveitis and the ocular symptoms were similar to those in HTLV-I uveitis in adults. The ocular inflammation responded to therapy with topical or systemic corticosteroids, but recurred in three patients. HTLV-I provirus DNA was detected by polymerase chain reaction (PCR) from infiltrating cells in the anterior chamber in one patient. The percentage of HTLV-I-infected cells in the peripheral blood mononuclear cells was measured by quantitative PCR, and the values were high (2.9 approximately 7.3%) in three cases tested as compared with an asymptomatic carrier. These five cases show that HTLV-I uveitis can be induced in a relatively short period (3 approximately 10 years) after the viral infection, and that HTLV-I uveitis should be considered as one possible etiology of uveitis in children, particularly in a viral endemic area.

Adolescent