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Biomedical subjects

M Kawashima

Publications and source records attributed to M Kawashima.

At least 109 records · Page 6Linked to original sources

A simple procedure for the isolation of rat kidney lysosomes.

BACKGROUND: A procedure for the isolation of highly purified lysosomes from normal rat kidney is described. METHODS: The method depends on the swelling of mitochondria when the postnuclear supernatant fraction is incubated with 2 mM Ca2+. The lysosomes can then be separated from the swollen mitochondria by Percoll density gradient centrifugation. RESULTS: The lysosomal fraction obtained by our method was enriched more than 30-fold in terms of marker enzymes with a yield of about 11%. Electron microscopic examination and the measurement of the activities of marker enzymes for various subcellular organelles indicated that our lysosomal preparation was essentially free from contamination by other organelles. CONCLUSION: We believe that this procedure for isolating kidney lysosome will be useful in the study of the mechanisms of specific modification, processing and catabolism of proteins.

Albumins↗

Clinical features of esophageal cancer in the octogenarian treated by definitive radiotherapy: a multi-institutional retrospective survey.

BACKGROUND: As age-related infirmity often influences treatment options and outcome of esophageal cancer, the optimization of treatment for the elderly, especially in octogenarians, has been the subject of considerable debate. METHODS: We performed a retrospective, multi-institutional survey to assess the effect of age on the outcome of definitive radiotherapy for esophageal cancer by a questionnaire sent to eight institutions in Japan. RESULTS: There were 362 evaluable replies. The patients included 317 males and 45 females, with a median age of 72 years (range 35-93 years), and 96% had squamous cell carcinoma. There were 30 clinical stage 1, 71 stage IIA, 17 stage IIB, 113 stage III and 116 stage IV cases. The stage was not specified in 16 cases. Multiple co-morbidities existed in 40% of the patients who were 70 years of age or older. There was no statistically significant age-related difference in the incidence of adverse reactions to radiotherapy (P > 0.05). Overall survival was more significantly affected by Karnofsky Performance Status than by the patient's age. The influence of performance status on cumulative survival for stage I and II disease was more pronounced in patients in their 80s. CONCLUSION: The safety of radiotherapy for esophageal carcinoma is not influenced by the patient's age. Because the performance status strongly influenced survival, the multi-disciplinary assessment of functional status is mandatory for optimizing the choice of treatment for patients in their 80s with esophageal cancer.

Adult↗

Arginine vasotocin receptor in the vagina of the oviduct of the hen.

The presence of receptor for arginine vasotocin (AVT) in the vagina of the oviduct of the hen was demonstrated by the use of radioligand binding assays on membrane fractions of the tissue. The binding to [125I]AVT was highly competitive with unlabeled AVT. Scatchard analysis revealed that the binding sites are of a single class. The equilibrium dissociation constant (Kd) was 0.48+/-0.05 nM (x+/-SEM; n = 6) in laying hens holding a hard-shelled egg in the uterus (shell gland) and 1.01+/-0.02 nM (n = 6) in nonlaying hens. The maximum binding capacity (Bmax) was 0.41+/-0.04 pmol/mg protein (n = 6) in laying hens and 0.81+/-0.01 pmol/mg protein (n = 6) in nonlaying hens. The Kd value of the laying hens varied from 0.39 to 1.20 nM during an oviposition cycle, showing an increase just prior to oviposition, and the Bmax value also varied from 0.30 to 0.66 pmol/mg protein, showing a gradual increase after 6 h prior to oviposition. In the nonlaying hen, both values were almost constant during a 24-h day. The changes in the binding affinity and capacity of AVT receptor of the vagina may be related to oviposition in the hen.

Animals↗

Pachyonychia congenita associated with steatocystoma multiplex.

We present an unique case of pachyonychia congenita associated with steatocystoma multiplex. A 33-year-old Japanese man had thickening and gray-brown dicoloration of all nails and a large number of nodules or tumors over his entire skin. No palmar and plantar hyperkeratosis, leukokeratosis of the mucous membranes, or follicular keratosis were observed. Histology of these tumors revealed the typical features of steatocystoma multiplex.

Adult↗

An immunohistochemical study of cadherin 5 (VE-cadherin) in vascular endothelial cells in placentas with gestosis.

OBJECTIVE: To evaluate vascular endothelial-cell functioning in placentas with gestosis, using the monoclonal antibody to cadherin 5. METHODS: The extra-cellular moiety of cadherin 5 was transfected into L-cells to enable us to examine their cell-adhesion activity. The expression of cadherin 5 was evaluated in human umbilical-vein endothelial cells and in placentas with gestosis by immunostaining using the anti-cadherin 5 antibody. A microspectrophotometric study also was conducted of the placentas with gestosis. RESULTS: We determined the total base sequence for cadherin 5 and found that it is homologous with a known cadherin but is a new, unique clone. Cadherin 5 has cell-adhesion activity and is expressed in endothelial cells at the cell-adhesion surface. The expression of cadherin 5 in endothelial cells took place in the placentas with gestosis, but to a lesser extent than in normal placental endothelial cells. CONCLUSIONS: The reduced expression of cadherin 5 in placentas with gestosis suggests that endothelial cell functioning is impaired in placentas with gestosis. Cadherin 5 in endothelial cells might influence placental functions and fetal development.

Amino Acid Sequence↗

Antisense oligonucleotides targeting c-fos mRNA inhibit rheumatoid synovial fibroblast proliferation.

OBJECTIVE: To determine whether antisense oligonucleotides targeting c-fos mRNA have the ability to inhibit the growth of interleukin 1 (IL1) stimulated fibroblast-like cells from the synovium in rheumatoid arthritis (RA). METHODS: Fibroblast-like cells established from RA synovium were stimulated by IL1 with antisense or sense oligonucleotides complementary to c-fos mRNA, and the proliferation of these cells was determined by 3H-thymidine incorporation. Effect of antisense oligonucleotides on expression of activator protein 1 (AP1) activity was evaluated using electrophoretic mobility shift assay. RESULTS: C-fos antisense oligonucleotides inhibited IL1 stimulated synovial fibroblast proliferation. The expression of AP1 activity induced by IL1 was suppressed by treatment with antisense oligonucleotides. CONCLUSION: These results suggest the feasibility of antisense strategies designed to suppress c-fos expression as therapeutic agents for RA.

Arthritis, Rheumatoid↗

Effect of tazarotene, an acetylenic retinoid, on human dermal fibroblast.

The inhibitory effect of tazarotene, an acetylenic retinoid, on human dermal fibroblast in vitro was compared to that of all-trans-retinoic acid. The proliferation of fibroblasts was inhibited by both retinoids at the concentration of 1 microM after 5 days of culture. Synthesis of DNA and collagen was inhibited by both retinoids concentration-dependently up to 10 microM, although tazarotene was weaker in the inhibition of collagen synthesis. These results suggest the possible usefulness of tazarotene in the treatment of fibrotic diseases.

Bromodeoxyuridine↗

Calcitonin receptor binding properties in bone and kidney of the chicken during the oviposition cycle.

The binding property of calcitonin (CT) in the membrane fraction of calvaria and kidney of egg-laying and nonlaying hens was analyzed using a [125I] CT binding assay system. Binding properties of CT receptors in both tissues satisfy the authentic criteria of a receptor-ligand interaction in terms of specificity, reversibility, and saturation. Scatchard plots revealed a single class of binding sites. Values of the equilibrium dissociation constant (Kd) and binding capacity (Bmax) in laying hens showed a decrease during the period between 3 h before and 2 h after oviposition. No change was observed in nonlaying hens. In vivo administration of 17beta-estradiol or progesterone caused the decrease in Kd and Bmax values. The results suggest that the binding affinity and capacity of the CT receptor in the calvaria and the kidney of the hen may be modulated by the ovarian steroid hormone.

Animals↗

Thiamine-deficient lactic acidosis with brain tumor treatment. Report of three cases.

Lactic acidosis due to thiamine deficiency is known to complicate chemotherapy and radiotherapy treatment of malignant extracranial tumors, but to the authors' knowledge, this complication has not been reported in patients treated for malignant brain tumors. They report three such cases, demonstrating that this complication can occur during treatment of brain tumors. In all patients, consciousness levels deteriorated within 1 to 2 days. Serum lactic acid levels increased to concentrations between 62 and 96.7 mg/dl, resulting in severe metabolic acidosis. A low blood thiamine level (9 ng/ml) was demonstrated at the onset in one case, and high-dose thiamine infusions dramatically improved lactic acidemia as well as impairment of consciousness in two cases. In the other case, hydrocephalus was suspected initially, resulting in a delay in thiamine supplementation. Clinical differentiation of this form of lactic acidosis from hydrocephalus or tumor progression can be very difficult in a patient undergoing treatment for a malignant brain tumor. Demand for thiamine is thought to be increased in patients with malignant brain tumors, and supplemental thiamine during treatment is necessary to prevent lactic acidosis. When this complication occurs, immediate treatment with sufficient thiamine is essential, together with normalization of pH by using sodium bicarbonate. With timely intervention, the level of consciousness can recover to the preacidotic state with no new neurological deficits.

Acidosis, Lactic↗

[An evaluation of efficacy of minocycline as an anti-rheumatic drug in patients with active and refractory rheumatoid arthritis].

The efficacy and safety of minocycline was investigated in Japanese patients with rheumatoid arthritis (RA) who had already received more than three disease modifying anti-rheumatic drugs (DMARDs). Minocycline was administered at 100 mg twice a day to fifteen patients with active RA. The drug efficacy was evaluated by the clinical variables including the number of painful and/or swollen joints, the duration of morning stiffness, grip strength, the erythrocyte sedimentation rate, serum concentrations of C-reactive protein, and the titer of rheumatoid factor. Three patients experienced adverse effects such as dizziness and abdominal pain or discomfort, but only one patient with abdominal pain and dizziness was discontinued. Fourteen RA patients, who had taken minocycline for at least 6 months, were subjected to the clinical evaluation. Among them, 8 patients (54%) showed a significant improvement of clinical valuables for disease activity, beginning even at 4 weeks of the therapy. The continued effects were observed in 8 patients with over 1 year-minocycline therapy. Intriguingly, an active patient with a history of multiple DMARDs-resistancy showed a marked favorable response to this drug. The present study indicates that minocycline may be an effective DMARD with highly safe performance for patients with active and refractory RA. This is the first demonstration of the benefit of minocycline in the Japanese patients.

Aged↗

[Desquamative interstitial pneumonitis accompanied by a variety of autoimmune abnormalities in an individual with a history of asbestos exposure].

A 75-year-old-man who had been occupationally exposed to asbestos over several decades was admitted complaining of dry cough and mild exertional dyspnea. Chest X-ray films revealed the presence of bilateral infiltrates. An open biopsy showed the lung pathology to be desquamative interstitial pneumonitis (DIP), and detected a calcified plaque of the left parietal pleura at cost-phrenic sulcus, which on microscopic inspection demonstrated thin needle-like structures suggestive of asbestos particles. A hormonal examination showed decreased levels of tri-iodothyronine and tetraiodothyronine and an increased level of thyroid stimulating hormone. Additionally, a positive microsome test indicated autoimmune hypothyroidism. A serological study was positive for anti-nuclear antibody and direct Coombs, and showed heightened levels of IgG. The patient was placed on oral prednisolone therapy; one month later, his symptoms had subsided, and his lung function and chest x-ray findings had improved significantly. In addition, the abnormal values for autoimmune and thyroid function eventually returned to normal ranges. These findings suggest that DIP and humoral immune dysfunction were caused by asbestos exposure, and responded well to steroid treatment.

Aged↗

Regulation of CD44-protein 4.1 interaction by Ca2+ and calmodulin. Implications for modulation of CD44-ankyrin interaction.

Erythrocyte membrane skeletal protein 4.1 isoforms have been identified in a variety of non-erythroid cells. However, interactions between protein 4.1 and its binding partners in non-erythroid cell membranes are poorly understood. In the erythrocyte membrane, protein 4.1 binds to the cytoplasmic domain of band 3 and, through this interaction, modulates ankyrin binding to band 3. The sequences LRRRY or IRRRY in band 3 mediate the interaction between band 3 and protein 4.1. The cytoplasmic domain of CD44, a transmembrane glycoprotein found in erythroid as well as non-erythroid cells, has internal sequences SRRRC and QKKKL. We wanted to determine if protein 4.1 binds to CD44 in a fashion analogous to its binding to band 3 and through this interaction modulates ankyrin binding to CD44. We report here that protein 4.1 binds to the cytoplasmic domain of CD44 with a dissociation constant on the order of 10(-7) M and that Ca2+ and calmodulin reduce the affinity of this interaction. Furthermore, although independent binding of both protein 4.1 and ankyrin to CD44 could be documented, binding of protein 4.1 prevented subsequent ankyrin binding. These studies have enabled us to identify a potentially important functional role for protein 4.1 in modulating ankyrin binding to CD44.

Ankyrins↗

The presence of mucosal human papillomavirus in Bowen's disease of the hands.

BACKGROUND: Bowen's disease (BD) of the genital skin region is generally associated with human papillomavirus (HPV) infection. Various molecular analyses have identified mainly HPV-16 in the lesions. However, the HPV genotypes associated with BD of the hands have not yet been characterized. METHODS: The skin specimens of 12 patients with BD of the hands were investigated clinicopathologically and immunohistochemically, and the total DNAs extracted from the skin were analyzed for the presence of HPV DNA using Southern blot hybridization and polymerase chain reaction (PCR) in combination with restriction fragment length polymorphisms (RFLP) and direct sequencing analysis of the amplified products. In addition, the histologic localization of HPV DNA was examined by in situ hybridization in paraffin embedded sections of HPV positive patients. RESULTS: In 8 of 12 BD lesions (66.7%), HPV types (HPV-16, -31, -54, -58, -61, -62, and -73) were detected by Southern blot hybridization and/or PCR with RFLP and direct sequencing analysis. In 6 of 7 HPV positive lesions examined (85.7%), the viral genomes were identified by in situ hybridization in the nuclei of keratinocytes in the upper stratum malpighii and/or stratum corneum. CONCLUSIONS: These results demonstrate that BD of the hands is frequently associated with HPV infection. The seven HPV genotypes are known as mucosal (genital) HPVs and to the authors' knowledge, this is the first time HPV-31, -54, -58, -61, -62, and -73 have been identified in BD lesions. These findings strongly suggest that HPVs related to mucosal lesions play an important role in the development of BD of the hands.

Adult↗

The role of granzyme B-expressing CD8-positive T cells in apoptosis of keratinocytes in lichen planus.

Epidermal basal cell injury with colloid body formation is a characteristic feature of lichen planus. Infiltrated cells are thought to be responsible for the epidermal injury. Ultrastructural findings of colloid bodies are typical of apoptosis. Granzymes in cytotoxic T lymphocytes are involved in apoptosis probably together with perforin. Based on this background, we analyzed the role of granzyme B in the mechanisms of epidermal injury in lichen planus. On electron microscopy, basal and suprabasal cells showed condensed chromatin and fragmented nuclei which are typical morphological features of apoptosis. Nuclei of colloid bodies were positively stained by the in situ nick end labeling technique indicating that colloid bodies are subsequently formed in the process of apoptosis. Immunohistochemical staining showed CD8-positive infiltrating cells to contain granzyme B. Cells undergoing exocytosis also contained granzyme B. By immunoelectron microscopy, granzyme B molecules were observed to be secreted from a lymphocyte to an apoptotic keratinocyte. These findings suggest that granzyme B-positive CD8 cells seem to induce apoptosis of keratinocytes in lichen planus.

Adult↗

Estrogen receptor is not primarily responsible for altered responsiveness of ovalbumin mRNA induction in the oviduct from genetically selected high- and low-albumen chicken lines.

The role of estrogen receptor on ovalbumin mRNA induction by steroid hormones was investigated in primary cultures of oviduct cells from estrogen-stimulated immature chicks of genetically selected high- and low-albumen egg laying lines (H- and L-lines). In experiment 1, the extent of ovalbumin mRNA induction and changes in estrogen and progesterone receptors were compared between the oviduct cells from H- and L-lines with or without steroid hormones in the culture medium. In experiment 2, the effect of estrogen receptor gene transfection on the induction of ovalbumin mRNA was studied in the oviduct cells from the L-line chicks. The results showed a close correlation of the changes in ovalbumin mRNA with the numbers of nuclear and total estrogen receptors in the oviduct cells but not with the numbers of nuclear and total progesterone receptors. Estrogen receptor gene transfection induced ovalbumin mRNA to a moderate extent in the absence of the steroid hormones. To our surprise, however, estrogen receptor gene transfection apparently suppressed the ovalbumin mRNA responsiveness to estrogen to a considerable extent. It was concluded, therefore, that the extent of estrogen receptor expression might not be primarily responsible for the differences in responsiveness to steroid hormones of oviduct cells from genetically selected H- and L-line chickens.

Animals↗

Technical considerations for fractionated stereotactic radiotherapy of hepatocellular carcinoma.

Technical aspects of fractionated stereotactic radiotherapy for solitary hepatocellular carcinoma have been investigated. Precise positioning of the patient and substantial reduction of the liver movement due to respiration were achieved by placing the patient ventrally on the treatment couch without a body cast. Repeated CT examinations were required for verification of tumor targeting. Though there were geometrical limitations on gantry rotation when the linac couch was rotated from its standard position, dose distributions obtained were found to be excellent. A patient with a small solitary lesion in the posterior segment of the liver received 52 Gy in 13 fractions over 29 days. He tolerated the treatment well without experiencing any morbidities or deterioration of liver functions. Three months later his alpha-fetoprotein value returned to normal and CT examinations revealed tumor shrinkage as well as a reduction in the viability of the tumor cells. The results suggest that it is possible to overcome technical difficulties associated with fractionated stereotactic radiotherapy of intraabdominal tumors.

Aged↗