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Biomedical subjects

M Kawano

Publications and source records attributed to M Kawano.

At least 145 records · Page 8Linked to original sources

Influence of serum follicle stimulating hormone to luteinizing hormone ratio during buserelin acetate-induced pituitary desensitization on ovarian response to exogenous gonadotrophins in an in-vitro fertilization and embryo transfer programme.

We investigated the effect of endogenous gonadotrophins during pituitary desensitization with gonadotrophin-releasing hormone agonist (GnRHa) on ovarian responsiveness or the outcome of in-vitro fertilization (IVF) and embryo transfer. The results of 67 women who participated in the IFV programme at Nagasaki University Hospital, Japan, were analysed retrospectively. All women received GnRHa from the third day of menstrual cycle, and ovarian stimulation with exogenous gonadotrophins was initiated when the serum oestradiol concentration decreased to < 30 pg/ml. The serum follicle stimulating hormone (FSH)/luteinizing hormone (LH) ratio, rather than serum FSH or LH concentrations during GnRHa-induced pituitary desensitization, showed a significant positive correlation with age and the total dose of exogenous gonadotrophins. The FSH/LH ratio also showed a significant negative correlation with oestradiol response and the number of retrieved oocytes, and was significantly lower in pregnant women compared with the non-pregnant group during pituitary desensitization. Our results indicate that, even under pituitary desensitization with GnRHa, the serum FSH/LH ratio influences individual ovarian responsiveness and the state of the intra-ovarian hormonal environment. Our results suggest that the FSH/LH ratio may be a useful clinical predictor of the ovarian response to exogenous gonadotrophins under pituitary desensitization.

Adult↗

Interaction between nucleocapsid protein (NP) and phosphoprotein (P) of human parainfluenza virus type 2: one of the two NP binding sites on P is essential for granule formation.

The paramyxovirus phospho- (P) and nucleocapsid (NP) proteins are involved in transcription and replication of the viral genome. To study the interaction between NP and P proteins, we established HeLa cell lines that constitutively expressed the NP and/or P proteins of human parainfluenza virus type 2 (hPIV-2). Co-immunoprecipitation assays revealed that the NP and P proteins can form complexes in HeLa cells expressing both proteins (HeLa-NP+P cells) and in mixed cell lysates of HeLa-NP and HeLa-P cells. Deletion mutant analysis of the P protein was performed to identify the regions of P protein that interact with NP protein. The results indicate that two independent NP-binding sites exist on P protein: one is located in the N-terminal part of the protein, aa 1-47, and the other in the C-terminal part, aa 357-395. In addition, cells co-expressing NP and P proteins with N-terminal deletions showed immunofluorescence staining patterns (granular pattern) similar to those found in hPIV-2-infected cells. However, cells co-expressing NP and P proteins with C-terminal deletions showed a different immunofluorescence staining pattern (diffuse pattern), indicating that the C-terminal region is required for granule formation.

Animals↗

Regulation of human immunodeficiency virus gp160-mediated cell fusion by antibodies against fusion regulatory protein 1.

We have isolated new MAbs directed against the human fusion regulatory protein 1 (FRP-1; CD98) molecule using human FRP-1-expressing L929 cells as antigens. The biological activities, and in particular the human immunodeficiency virus (HIV)-mediated fusion regulatory activity of seven anti-FRP-1/CD98 MAbs were analysed using the U937/gp160 cell line, which is a CD4+ U937 cell line expressing HIV gp160. Two MAbs induced multinucleated giant cell formation in U937/gp160 cells and the other five MAbs showed no fusion-inducing ability. However, four of these MAbs suppressed multinucleated giant cell formation of U937/gp160 cells induced by the activating anti-FRP-1 MAbs. Interestingly, five of the MAbs induced multinucleated giant cells in peripheral blood monocytes and one MAb showing fusion-inducing ability in U937/gp160 cells suppressed multinucleated giant cell formation of monocytes induced by anti-FRP-1 MAbs. Furthermore, four of the anti-FRP-1 MAbs suppressed cell fusion of Jurkat/gp160 cells, which are Jurkat cells expressing HIV gp160. Thus, FRP-1/CD98 is capable of either activating or inhibiting HIV-mediated cell fusion depending on whether an enhancing or inhibiting antibody is used, indicating that FRP-1/CD98 is a multipotential molecule. Thus, HIV-mediated cell fusion can be regulated by modification of the FRP-1 system. Furthermore, the present study demonstrates that the FRP-1 and FRP-2 systems are interdependent.

Animals↗

Identification of the sequences responsible for nuclear targeting of the V protein of human parainfluenza virus type 2.

In human parainfluenza virus type 2 (hPIV-2)-infected cells, anti-phosphoprotein (P)-specific monoclonal antibody (MAb) densely stained the perinuclear regions of infected cells throughout infection, indicating that the P protein was localized exclusively in the cell cytoplasm. By contrast, antigens recognized by MAbs directed against the P-V-common domain of hPIV-2 were located predominantly in the cytoplasm, but in some hPIV-2-infected cells they were also found in the nuclei, suggesting that a fraction of hPIV-2 V protein is localized there. hPIV-2 V protein expressed from a cDNA clone was localized in the nuclei of transfected cells. By using indirect immunofluorescence analyses, we examined the intracellular localization of various sequentially deleted V proteins, to determine the nuclear localization signals (NLS) of the V protein. Two noncontiguous regions in the V protein were required for nuclear localization and retention, since deletion of these regions [region I (aa 1-46) and region II (aa 175-196)] resulted in cytoplasmic localization. Both regions resulted in nuclear localization independently. A nucleoplasmin-like NLS was identified in region II but no consensus targeting sequence could be found in region I. When NP protein was co-expressed with V protein or the N-terminal fragment (aa 1-46) of V protein, a fraction of the NP protein was translocated into cell nuclei.

Amino Acid Sequence↗

Granulomatous cholangitis in chronic hepatitis C: a new diagnostic problem in liver pathology.

A case of chronic hepatitis C at the pre-cirrhotic stage complicated with hepatocellular carcinoma is reported. The patient, a 64 year old female, showed elevated levels of serum alkaline phosphatase and immunoglobulin M. Antimitochondrial antibodies were negative by indirect immunofluorescence. Western blotting using beef heart mitochondria and recombinant polypeptides coding for mitochondrial antigens revealed that the patient's serum was positive only for the E2-subunit of the branched chain ketoacid dehydrogenase complex. In the non-neoplastic liver, chronic non-suppurative cholangitis surrounded by epithelioid granuloma, resembling the granulomatous destructive cholangitis of primary biliary cirrhosis, was found. The damaged bile ducts were immunohistochemically minimally positive or ambiguous for HLA-DR, and their expression of the E2-subunit of the pyruvate dehydrogenase complex E2 (PDC-E2) was diffuse or granular, and not typical of primary biliary cirrhosis. There was no bile duct loss, and orcein-positive copper binding granules reflecting chronic cholestasis were negative in periportal hepatocytes. The overall features in this case were consistent with primary biliary cirrhosis presenting an infrequent profile of antimitochondrial antibodies and atypical expression of HLA-DR and PDC-E2 on biliary epithelial cells, with late superimposition on chronic hepatitis C. However, it is also possible that this is a case of chronic hepatitis C with hepatitis-associated bile duct damage accompanied by granulomatous reaction. Either way, this case raises new diagnostic issues in the differential diagnosis of chronic liver diseases presented with granulomatous cholangitis.

Aged↗

Clinical evaluation of orthodontic treatment after surgical repositioning or transplantation of teeth.

Fourteen impacted teeth in thirteen patients aged 9 to 17 years were either repositioned or transplanted, followed by postoperative orthodontic treatment. Successful results were achieved in twelve of the teeth. In one tooth, ankylosis of the root prevented a successful outcome, and deficient bone healing of the alveolar process led to an unsuccessful result in a second tooth.

Adolescent↗

[The effectiveness of neoadjuvant chemotherapy for invasive bladder cancer].

BACKGROUND: Sixteen patients with invasive bladder cancer were received neoadjuvant methotrexate, vinblastine, pirarubicin and cisplatin chemotherapy on our planning protocol. METHODS: The tumor was evaluated after 1 course of chemotherapy by radiographic examination, urine cytology, cystoscopy and random biopsy. If the response is CR or PR, one more course of chemotherapy was performed, and cystectomy was carried out. If the response is NC or PD, cystectomy was immediately carried out. Twelve of them were underwent cystectomy and four were preserved bladder. Clinical response was evaluated by echo, CT, MRI, urinary cytology, cystoscopy and random biopsy. RESULTS: Clinical CR was observed in 25% and PR was 37.5%. Pathological CR was observed in 31.3% and PR was 37.5%. The different rate between clinical and pathological evaluations was 31.3% and the result suggests that we should find the method of more accurate staging evaluation. Four patients who were evaluated clinical CR were selected bladder-preserving. However, two of them (50%) had recurred; one had grade 3 tumor was treated by total cystectomy and the other had multiple tumors was treated by one course of M-VAC and TUR-Bt. CONCLUSION: We should consider which cases are possible to preserve bladder by investigating the tumor characteristics.

Aged↗

[Classification of patterns of nocturnal penile tumescence with continuous monitoring of penile rigidity: analysis of various factors affecting erection].

In 43 men who required further examination of erectile function, we analyzed the relationship between the various factors including hormone levels, penile blood pressure index (PBPI), shape of the arterial wave, erection period after prostaglandin E1 injection and dynamic cavernosography/metry, as well as the patterns of nocturnal penile tumescence (NPT) with rigidity. Normal, dissociation, short episode, low amplitude and flat trace patterns, which were classified by Kaneko, were observed in 11, 8, 11, 2 and 11 men, respectively. Uncoupling was not observed in any men. There were no differences in the levels of prolactin and LH among the 6 patterns. A high FSH level was seen in men with a flat trace pattern; a low level of free testosterone ( < 12.2 pg/ml) was seen in men with dissociation, low amplitude and flat trace patterns, and low PBPI ( < 0.75) in men with flat trace pattern; and an arteriosclerotic shape of the arterial wave was seen in men with short episode and flat trace patterns. Each erection period immediately after prostaglandin E1 injection in men with dissociation, short episode, low amplitude and flat trace was shorter compared with cases with a normal pattern. Venoocclusive dysfunction was observed in 5 men with a dissociation pattern and 6 men with a flat trace pattern. In conclusion, an integral analysis of pattern of NPT with rigidity and other examinations are useful for differentiating organic impotence from psychogenic impotence and for accurately diagnosing pathophysiological disorders of organic impotence and underlying diseases.

Adult↗

[A case of oligozoospermia with marginal hyperprolactinemia due to pituitary microadenoma].

We present an infertility male with prolactin level of 37.5 ng/ml. The patient had pituitary microadenoma detected by Gd-DTPA enhanced magnetic resonance imaging (MRI). After bromocriptine was administrated for 4 months, the size of microadenoma decreased, and sperm density and mortality improved. His wife became pregnant after 6 months. Dynamic MRI is a useful modality for detection of pituitary microadenoma, and bromocriptine is also useful for treatment of oligospermic patients with marginal hyperprolactinemia.

Adenoma↗

Intracellular localization of antigens recognized by anti-vimentin monoclonal antibodies (mAbs): cross-reactivities of anti-vimentin mAbs with other cellular components.

Monoclonal antibodies were raised against immunoaffinity-purified fusion regulatory protein (FRP)-1 complex from membrane fraction of HeLa cells. Immunoblotting and immunoprecipitation studies showed all ten antibodies reacted with a 55 kDa band of cell lysate and purified vimentin. Interestingly, one of the antibodies (mAb57) cross-reacted with purified tropomyosin and myosin. Further analyses using vimentin chemically cleaved by 2-nitro-5-thio-cyanobenzoic acid, and lambda gt 11 cDNA which encoded a partial sequence of vimentin indicated that six mAbs recognized epitopes between amino acids 1 and 313 and the other four mAbs recognized epitopes in the area between residues 314 and 326. Indirect immunofluorescence microscopy using 3% formalin-fixed, 0.1% Triton X-100 treated HeLa cells revealed that seven antibodies stained various intracellular components other than vimentin, while three antibodies stained vimentin filaments alone. Furthermore, flow cytometric analysis showed one of the antibodies (mAb25) clearly stained the surface of unfixed HeLa cells. All immunofluorescent findings were the same when HeLa, baby hamster kidney (BHK) and murine L229 cells were examined. These results indicate that we could obtain unique anti-vimentin mAbs which show cross-reactivities with previously undescribed cell surface and intracellular molecules including tropomyosin and myosin. Taken together, there are two possibilities that explain our findings: (1) The unknown molecules may have structural similarity to vimentin. (2) Our anti-vimentin mAbs can react specifically with structurally distinct epitopes present on both unknown molecules and vimentin. In either case, our cross-reactive mAbs, which recognized undescribed epitopes on vimentin, maybe provide useful tools for studying intermediate filaments and related cellular components.

Animals↗

[Pulmonary and endobronchial tuberculosis with subclavian artery-pulmonary artery shunts].

A 77-year-old woman with a productive cough and fever was admitted to the hospital. Pulmonary and endobronchial tuberculosis, pneumonia of the left upper lobe, and stenosis of the left main bronchus were diagnosed. She was given the antimycobacterial drugs isoniazid, rifampin, and streptomycin, and her condition improved. Two months later, bronchoscopy revealed semilunar-shaped stenosis of the left main bronchus, and auscultation revealed wheezing in the middle-end expiratory phase. A continuous flow murmur (Levine III) was also heard at the left anterior chest wall. Cardiac catheterization with subclavian arteriography revealed two left subclavian-pulmonary shunts. In a case of systemic-pulmonary shunt such as this, the bronchial stenosis could be surgically repaired, but the result would be an increase in dead space. If left untreated, the pulmonary hypertension would progress and symptoms of pulmonary disease would become more severe. Subclavian-pulmonary artery shunt is a very rare complication of pulmonary tuberculosis. Surgical treatment should consist of open bronchoplasty along with lobectomy and removal of the shunt, rather than embolization of the shunt and endoscopic bronchoplasty.

Aged↗

[Glucose intolerance after percutaneous ethanol injection therapy (PEI) in liver cirrhosis patients with hepatocellular carcinoma].

We studied the effect of percutaneous ethanol injection therapy (PEI) on glucose tolerance in liver cirrhosis patients with hepatocellular carcinoma. All of 10 patients underwent PEI and aspiration biopsy of the tumor on separate day. Two-time oral glucose tolerance tests (OGTT), before and after PEI, were performed in all patients. There were no significant changes in blood glucose and insulin chronologically measured on aspiration biopsy and PEI. To detect changes in glucose tolerance, we compared the results of OGTT before PEI with those of OGTT after PEI. On the basis of results of OGTT before PEI, patients were classified to impaired glucose tolerance group (4) and diabetes mellitus group (6). Blood glucose at 180 minutes on OGTT after PEI showed significantly higher value than that of OGTT before PEI, but insulin response was not suppressed. From these experiments we speculate that exaggerated insulin resistance due to injected ethanol may be one of the factors influencing glucose tolerance after PEI.

Aged↗

Identification of regions on the hemagglutinin-neuraminidase protein of human parainfluenza virus type 2 important for promoting cell fusion.

The hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins of two paramyxoviruses, human parainfluenza virus type 2 (PIV2) and simian virus 41 (SV41), were expressed in HeLa cells by transfecting with recombinant plasmid harboring each glycoprotein gene. Expressed F proteins could not induce cell fusion by themselves, but evoked prominent cell fusion when coexpressed with homologous HN proteins. It was also proved that PIV2 HN protein could weakly promote SV41 F-mediated cell fusion. By analyzing the fusion-promoting function of chimeric HN proteins of PIV2 and SV41, it was revealed that the N-terminal region (about 16% of total amino acids) of either PIV2 HN or SV41 HN protein could define the type-specific fusion-promoting function for homologous F protein. Analyses of additional chimeras indicated that the N-terminal region in PIV2 HN protein (designated region I, consisting of 94 amino acids) could be reduced to a 58-amino-acid region (region I') which was located at the membrane-proximal end of the ectodomain. Furthermore, PIV2 HN protein proved to promote cell fusion mediated by PIV4A F protein. Unexpectedly, analyses of another set of chimeras revealed that the promoting function of PIV2 HN protein for PIV4A F-mediated cell fusion was not merely carried by its region I but also by another region ranging from residue 148 to 209 (region II). Finally, it was indicated that regions I' (in the presumed stalk domain) and II (in the globular head) in PIV2 HN protein might play important roles in promoting cell fusion mediated by the F proteins.

Amino Acid Sequence↗

Molecular characterization of fusion regulatory protein-1 (FRP-1) that induces multinucleated giant cell formation of monocytes and HIV gp160-mediated cell fusion. FRP-1 and 4F2/CD98 are identical molecules.

Fusion regulatory protein (FRP)-1 regulates virus-mediated cell fusion and fusion of monocytes. Eleven of fifteen N-terminal amino acids of FRP-1 were the same as the amino acid sequence of 4F2/CD98 heavy chain. FRP-1 molecules were detected in Con A- or IL-2-stimulated lymphocytes, while FRP-1 was rare on resting lymphocytes. These properties of FRP-1 are similar to those of 4F2/CD98. Treatment of monocytes with anti-4F2/CD98 mAbs resulted in cell fusion, and other mAbs directed against 4F2/CD98 induced formation of multinucleated giant cells of Cd+U2ME-7 cells, a CD4+U937 cell line transfected with the HIV gp160 gene. Both anti-4F2/CD98 and anti-FRP-1 mAbs reacted with murine L929 cells expressing human 4F2/CD98 transiently or constitutively. When Newcastle disease virus (NDV)-infected L929 cells expressing human FRP-1/CD98 were incubated with mAb 4-5-1, an anti-FRP-1 mAb, multinucleated giant cells were induced; thus, FRP-1/CD98 molecules expressed in L929 cells are functional for fusion regulatory activity.

Amino Acid Sequence↗

A cell fusion-inhibiting monoclonal antibody binds to the presumed stalk domain of the human parainfluenza type 2 virus hemagglutinin-neuraminidase protein.

Previously, we obtained a neutralizing monoclonal antibody directed against the hemagglutinin-neuraminidase (HN) protein of human parainfluenza type 2 virus (PIV2), which was able to prevent cell fusion without affecting the hemagglutinating and neuraminidase activities. In this study, four escape mutants of PIV2 have been obtained under pressure of the monoclonal antibody. Intriguingly, the HN protein of each mutant proved to have two amino acid substitutions, one of which is at 83Asn or 91Lys, and another one is at 150Leu, 160Ala, or 186Met. One mutant designated F13, which has substitutions at 83Asn and 186Met in the HN protein, could not cause cell fusion in HeLa cells despite its multiple replication, while the other mutants formed typical syncytial cells. The deduced amino acid sequence of F13 fusion (F) protein proved to be identical to that of wild-type F protein, and furthermore, protein expression analyses have revealed that the low-fusion phenotype of F13 was due to its mutated HN protein, whose antigenicity to the monoclonal antibody was abolished by the single mutation at 83Asn. These observations have suggested that the principal epitope for the monoclonal antibody resides in the presumed stalk domain of the HN protein, which may play an important role in promoting cell fusion.

Amino Acid Sequence↗