Search PubMed⌕ Search

Biomedical subjects

M Kanisawa

Publications and source records attributed to M Kanisawa.

At least 37 records · Page 2Linked to original sources
PubMed1995

Expression of connexin 32 gap junction protein in the kidneys during fetal development of the hamster (Mesocricetus auratus).

The expression of gap junction protein was examined immunohistochemically using affinity-purified antibody against rat liver gap junction protein, connexin 32 (Cx32), in the kidneys of fetal (gestation days 13-16) and adult Syrian golden hamsters. Phalloidin histochemical staining, PNA- and RCA I-lectin staining, NCAM immunostaining, and alkaline phosphatase and Na(+)-K(+)-ATPase enzyme-histochemical staining were performed in combination with Cx32 immunostaining. The kidney sections were observed with a confocal scanning laser microscope. By gestation day 13, Cx32 immunoreactivity was observed in the differentiating tubules. The Cx32 staining was localized on the lateral cell membrane of the cells lining the developing proximal tubules, while the S-shaped bodies, developing distal tubules, and collecting tubules showed no positive immunostaining. As the kidney developed, the density of Cx32 immunoreactivity increased. As the gap junction provides pathways for cell-cell communication, the development of Cx32 expression may imply that this structure plays an important role in renal tubule development. Confocal scanning laser microscopy provided a clear image of the fluorescence-labeled cell structures, free from out-of-focus blur. Using the same sections, stereoscopic images were easily reconstructed from serial optical sections, and were helpful in understanding the spatial distribution of Cx32 expression in the developing fetal proximal tubules.

Animals↗
PubMed1995

Hamster pulmonary endocrine cells with neural cell adhesion molecule (NCAM) immunostaining.

Pulmonary endocrine cells of Syrian golden hamster were stained for neural cell adhesion molecule (NCAM) with indirect fluorescent immunostaining and observed with a confocal laser scanning microscope equipped with an argon laser. Sections 100 microns thick of hamster lung fixed with phosphate-buffered 4% paraformaldehyde were prepared. The sections were incubated with rat monoclonal antibody against NCAM, followed by fluorescence-labeled antibody against rat immunoglobulin. Some were doubly immunostained for NCAM and one of the following endocrine markers: neuron-specific enolase, calcitonin gene-related peptide and serotonin. Expression of NCAM in the hamster airway epithelium was seen in cell nests resembling neuroepithelial bodies (NEBs). NCAM immunostaining was positive at the lateral cell borders between the cells composing the nest, but negative at the border with the adjacent, presumably non-endocrine cells. Double immunostaining confirmed that the grouped cells with NCAM immunoreactivity were of an endocrine nature, but that single endocrine cells did not show NCAM immunoreactivity. An electron microscopic study with NCAM immunostaining confirmed the light microscopic study. These suggest that NCAM expression could be important for the morphogenesis of NEBs. A confocal laser microscope was used to make three-dimensional images of NEBs after NCAM immunostaining and the spatial interaction between NEBs and the surrounding microenvironment was studied.

Animals↗
PubMed1995

Alkaline phosphatase reactivity in rabbit airway epithelium: a potentially useful marker for airway basal cells.

The purpose of this study was to investigate alkaline phosphatase (ALPase) reactivity in rabbit airway epithelial cells. Acetone-fixed, methyl benzoate and xylene-cleared (AMeX-treated) paraffin sections of trachea, bronchus, and lung tissue were stained by an azo dye coupling method for ALPase and examined by light microscopy. Electron histochemical staining was also performed in order to study the sensitivity and specificity of reactivity in each cell type. ALPase reactivity at the light microscopic level was observed exclusively in tracheo-bronchial basal cells, and not in bronchiolar basal cells. By electron microscopy, ALPase reactivity was noted in 97.9% of basal cells in the trachea, 97.0% of basal cells in the bronchus, and 94.5% of basal cells and 15.4% of Clara cells in the bronchiole. This was also true for dispersed tracheal epithelial cells. Reactivity was rarely observed in ciliated cells, non-goblet-type secretory cells, and undetermined cells. The reactivity was heat-labile, levamisole-sensitive, and of a non-specific type. These findings indicate that basal cells of rabbit trachea and bonchus have fairly high specificity for ALPase of a non-specific isozyme (92.2% and 95.6%, respectively). Therefore, ALPase is considered to be a useful marker for these cells.

Alkaline Phosphatase↗
PubMed1995

Morphologic alterations and cytokinetic studies of tracheal autograft epithelium in rabbits.

BACKGROUND: Although tracheobronchoplasty has been used widely in the field of thoracic surgery, few details of the morphologic changes in and cytokinetics of the graft epithelium have been reported. The aim of this study was to focus on these aspects in autografted rabbit tracheas. METHODS: Resected cervical tracheas were anastomosed immediately after removal, retrieved on postoperative days 1 through 28, and examined morphologically. Mitotic and bromodeoxyuridine-labeling indices of the graft epithelium were analyzed. RESULTS: On postoperative days 1 to 4, the graft epithelium showed focal desquamation at the anastomoses. Ciliated cells disappeared during postoperative days 4 to 7 and then increased gradually. Nonciliated cells retained a somewhat columnar shape on postoperative days 4 to 7, except at denuded foci. Thereafter, the grafts were covered completely with pseudostratified mucociliary epithelium. On postoperative day 4, both indices were maximal and appeared higher at the anastomotic than midgraft sites. CONCLUSIONS: Most of the graft epithelium was preserved during acute ischemia and then started to regenerate. The increased regenerative activity near the anastomoses may be attributable to mechanical damage or different nutritional conditions.

Anastomosis, Surgical↗
PubMed1995

Bromodeoxyuridine and iododeoxyuridine double immunostaining for epoxy resin sections.

To establish bromodeoxyuridine (BrdUrd)/iododeoxyuridine (IdUrd) double immunostaining for thick sections of epoxy resin-embedded tissues, young hamsters received intra-peritoneal injections of IdUrd and BrdUrd 3 hr and 1 hr before sacrifice, respectively. The intestines were fixed with phosphate-buffered 4% paraformaldehyde and embedded in an Epon-Araldite mixture. The epoxy resin was removed completely by a sodium methoxide/benzene/methanol solution. This epoxy resin removal method was effective for BrdUrd/IdUrd immunostaining using a mono-specific antibody for BrdUrd (Br-3) and a bi-specific antibody for BrdUrd (Br-3) and a bi-specific antibody for BrdUrd and IdUrd (IU-4), followed by the ABC complex method. Epoxy sections stained with these antibodies showed clear localization of nuclei incorporating the two thymidine analogues with precise morphology of labeled cells. Furthermore, ultrastructural observation of thin sections adjacent to thick sections immunostained for BrdUrd/IdUrd confirmed the cell type and ultrastructural features of cells labeled with these thymidine analogues.

Animals↗
PubMed1995

Proliferative potential and p53 overexpression in precursor and early stage lesions of bronchioloalveolar lung carcinoma.

To elucidate the pathogenesis of bronchioloalveolar lung carcinoma (BAC), we evaluated the lesion size, growth fraction, and p53 overexpression of atypical adenomatous hyperplasia (AAH) and early stage BAC. AAH was classified as showing low grade or high grade atypia. AAH-like carcinoma, presumably very early stage BAC, was distinguished from AAH in that it exhibited remarkable atypia suggestive of malignant potential and from overt BAC in that it lacked unequivocal malignant features, including invasive/destructive growth. The growth fraction was determined immunohistochemically in terms of the Ki-67 labeling index. The overexpression of p53 was evaluated by assessing the nuclear accumulation of immunoreactive p53 protein. Both the lesion size and the growth fraction increased from low grade AAH, to high grade AAH, to AAH-like carcinoma, and to overt adenocarcinoma. The overexpression of p53 in AAH-like carcinoma was similar to that in overt adenocarcinoma and was more frequent than that in AAH. Our findings indicate that AAH, AAH-like carcinoma, and overt BAC represent different categories, although the cellular events occurring in these lesions presumably represent a continuous spectrum of the changes that are reflected in the cytomorphology and lesion size. The findings here suggest that AAH and AAH-like carcinomas constitute a population of heterogeneous lesions representing different steps toward overt BAC.

Adenocarcinoma, Bronchiolo-Alveolar↗
PubMed1995

[A combined in vitro and in vivo system for culture of epithelial cells from distal human airways].

A system for combined in vitro and in vivo culture of epithelial cells from distal human airways was established. Lung tissues that appeared to be generally normal were obtained from lungs removed surgically from patients with lung cancer. Small pieces of peripheral lung tissue were placed on culture dishes and cultured in F-12 complete medium containing serum and various growth factors, to obtain outgrown cells. For in vivo culture, rat tracheal grafts were de-epithelialized by freezing and thawing and were then used as culture vessels. Outgrown cells were harvested after 4 weeks of in vitro culture, inoculated into the denuded tracheal grafts, and then implanted into nude mice. For comparative purposes, bronchial fragments were also cultured in vitro and in vivo, by the same method. In vitro efficiency of colony formation was about the same for cells derived from peripheral lung tissue and from bronchial tissue (14.8 +/- 8.9% and 16.0 +/- 4.7%, respectively). Four weeks after implantation, the grafts were retrieved and processed for morphologic evaluation. By that time, grafts in both groups had totally re-epithelialized. Therefore, the growth potential of the cells derived from peripheral lung tissue and from bronchi in vivo appeared to be almost the same. Newly formed epithelial cells in grafts showed the same well-developed pseudostratified columnar form in both groups at 4 weeks. The time course of epithelial cell differentiation was also studied, with outgrown cells from lungs. Two days after implantation, undifferentiated cells were attached to the inner surface of the grafts as a single cell layer, and at 4 days, small cell nests containing mitotic cells were observed. At 1 week, the grafts were totally covered with undifferentiated cells. Over 2 to 3 weeks, differentiated cells (ciliated, secretory, and basal cells) appeared, and the epithelia had become fully developed by 4 weeks. As reported previously, cells that outgrew from lung explants were considered to be derived from bronchioles. Therefore, this system may be useful for studies of growth and differentiation of human bronchiolar epithelial cells under various conditions.

Animals↗
PubMed1994

Significance of hepatocellular proliferation in the development of hepatocellular carcinoma from anti-hepatitis C virus-positive cirrhotic patients.

BACKGROUND: There is a hypothesis explaining the pathogenesis of carcinoma that increased proliferation of tissue cells correlates with the development of carcinoma, presumably by increased rate of random mutations and by promotion. In this study, the significance of hepatocellular proliferation in the development of human hepatocellular carcinoma (HCC) from anti-hepatitis C virus (HCV)-positive cirrhotic patients was studied. METHODS: Twenty-eight Child A cirrhotic patients who were anti-HCV (C-100 antibody) positive were studied. At the beginning of the study, the in vitro uptake of bromodeoxyuridine (BrdU, a thymidine analogue) by hepatocytes in biopsied liver specimens was investigated as labeling indices (LIs), and they were divided into high-DNA synthetic (BrdU LI > or = 1.5%) and low-DNA synthetic (BrdU LI < 1.5%) groups. The patients were then surveyed prospectively with frequent ultrasonography (every 3 months) for the development of HCC for 3 years. RESULTS: The mean BrdU LI plus or minus standard deviation for 14 cirrhotic patients with high-DNA synthesis activity (BrdU LI > or = 1.5%) was 2.7 +/- 0.8%, and this was significantly (P < 0.001) higher than that for 14 cirrhotic patients with low-DNA synthesis activity (BrdU LI < 1.5%, 0.5 +/- 0.3%). Nine of 14 (64.3%) of the cirrhotic patients with high-DNA synthesis activity developed HCC in the 3-year period, in contrast to only 2 of 14 (14.3%) of the cirrhotic patients with low-DNA synthesis activity P < 0.05).

Aged↗
PubMed1994

Effects of vitamin A on proliferation of human distal airway epithelial cells in culture.

Using a serum-free culture method, we investigated the effects of vitamin A on the proliferation of human distal airway epithelial cells. Outgrowth of epithelial cells from lung tissue explants was enhanced by treatment with all-trans retinol at concentrations of 10(-8) to 10(-7) M. The colony-forming activity of cells harvested from the primary culture and replated onto Swiss 3T3 fibroblastic feeders was, in contrast, significantly reduced by 10(-7) M to 10(-5) M retinol. When the primary cells were harvested and subcultured on Primaria plates, population expansion was also inhibited by retinol at 10(-10) to 10(-6) M. We further investigated the cells to determine whether there was any difference in sensitivity to the growth-inhibitory effects of vitamin A between cells from the primary culture incubated with and without retinol. The population increase in cells harvested from the primary culture was inhibited equally in retinol-treated and non-treated cells by subsequent treatment with retinol or retinoic acid, this inhibition being dose-dependent. DNA synthetic activity was also inhibited. Interestingly, both the growth rate and the colony-forming efficiency on feeders were greater in the subculture of cells from the retinol-treated primary culture than in those non-treated. When the cells in the secondary subculture were treated with retinoic acid and replated again, they showed a greater population increase rate than those non-treated. Our results showed that human distal airway epithelial cells isolated from lung tissue were sensitive to the growth-inhibitory effect of vitamin A, but the proliferative potential in some fraction of the epithelial cell population was possibly enhanced by vitamin A treatment.

3T3 Cells↗
PubMed1994

Haemopoietic spleen colony formation in the rat: effect of 89Sr-induced bone marrow aplasia.

Regulation of spleen-colony formation, a clonal assay for haemopoietic stem cells, is very different in mice and rats. In the rat, there is an involution of the colony-forming ability of the spleen during infantile development, whereas in mice the ability is maintained throughout life. In our re-evaluation of endogenous spleen colony formation in rats after graded doses of total-body irradiation, colonies ceased to appear after 12 weeks of age. However, histological sections showed that there were tiny colonies growing in the spleen, even at 20 weeks. These microscopical colonies developed into visible colonies when the rats were given treatments that increased the haemopoietic requirement. Thus, the amount of 59FeCl3 incorporated into the spleen increased to compensate for a decrease in uptake by the bone marrow. Rats in which bone-marrow activity was inhibited by 89SrCl2, showed extensive colony formation in the spleen, even after 12 weeks of age, when the endogenous colonies were induced by a sublethal dose of radiation. About twice as much 59Fe acetate activity was incorporated into the spleens of the experimental animals than in those of control rats. These findings imply that spleen-colony formation responds to the haemopoietic requirement of the spleen rather than that of the bone marrow. Furthermore, the requirement of the spleen seems to be much smaller in rats than in mice, because bone-marrow capacity for haemopoiesis is relatively larger in rats.

Aging↗
PubMed1994

Airway epithelial cell changes in rats exposed to 0.25 ppm ozone for 20 months.

The present study was designed to characterize and quantify morphologic changes occurring in rat intra-pulmonary airway epithelia after long-term exposure to a high ambient level of ozone. Fifteen rats were exposed to 0.25 ppm ozone for 20 months (5 hr/day, 7 days/week) and a control group of 15 rats were exposed to filtered room air. Many intra-cellular brown pigmented granules (presumed to be lipofuscin) were seen in both the control and exposed animals; however, more of these granules were observed in the bronchial and bronchiolar epithelia of the exposed animals. To detect DNA synthesis that occurred in airway epithelial cells during the repair process induced by ozone toxicity, bromodeoxyuridine (BrdU) was injected intraperitoneally one hour before animals were sacrificed; the BrdU-labeling index was evaluated immunohistochemically. There was no difference in the BrdU-labeling index between the groups, the airway epithelia of the ozone-exposed animals exhibiting tolerance to ozone toxicity. However, the epithelial populations in the airways were altered by ozone exposure, and regional differences were seen in the changes. In the exposed animals, mucous cells increased in the lobar bronchus. Calcitonin gene-related peptide-immunoreactive pulmonary endocrine cells and neuroepithelial bodies were more frequently observed in the terminal bronchioles of the exposed animals than in the controls, although there were no differences in the lobar bronchus. Moreover, one adenoma in the peripheral lung was found in the exposed animals, while no tumorous lesions were detected in the controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenoma↗
PubMed1993

Hemopoietic stem-cell compartment of the SCID mouse: double-exponential survival curve after gamma irradiation.

It has been reported that SCID (severe combined immunodeficiency, scid/scid) mice are more radiosensitive than normal mice. In the present studies, graded doses of radiation were given to bone marrow cells from SCID mice, and double-exponential survival curves were observed for day-9 and day-12 colony-forming units in the spleen (CFU-S). Single-exponential curves were found for SCID CFU in in vitro assays for granulocyte/macrophage-CFUs and erythroid burst-forming units, as reported elsewhere. Since the size of this more resistant fraction seems to decrease with stem-cell maturation, the finding implies that this fraction is a primitive subpopulation of the stem-cell compartment. The mean lethal dose (D0), however, of this less sensitive SCID CFU-S is much less than the D0 of regular CFU-S in normal littermates. Spleen colonies produced by SCID bone marrow were relatively small and abortive. The size of these colonies decreased nearly exponentially with increasing doses of radiation. These colonies were believed to be produced by this less sensitive fraction of the stem cells, which carried residual injuries. The colonies produced by the sensitive fraction have disappeared, being killed by a relatively low dose of radiation. This observation might account for the high lymphomagenesis arising from primitive hemopoietic stem cells in SCID mice, because the smallness of the colonies suggests that there is unrepaired or misrepaired damage. Furthermore, this less sensitive fraction might be a source of the "leaky" change of T and B cells, possibly due to the induction of an equivocal repair system which appears in the later stages of life in the SCID mice.

Animals↗
PubMed1993

An immunohistochemical study of adenoid cystic carcinoma of the external auditory canal.

Specimens from five cases of adenoid cystic carcinoma of the external auditory canal were studied by immunohistochemical staining, and findings were compared with those from adjacent non-neoplastic tissues containing ceruminous glands. In the ceruminous gland, cytokeratin showed diffuse positive staining, while myoepithelial cells were stained for smooth muscle actin, desmin, S-100 protein and vimentin. The epithelial markers used were cytokeratin, carcinoembryonic antigen and secretory component and stained at various densities the inner cells of the tubular component and duct-lining cells in the cribriform component of tumor tissues. In contrast, the muscular markers, smooth muscle actin and desmin, and the mesenchymal marker, vimentin, were positive in the outer cells of the tubular component and in the cyst-lining cells of the cribriform component. S-100 protein immunoreactivity showed paradoxical results; positive findings occurred in the myoepithelial cells of the ceruminous glands and in the inner cells of the tubular component and duct-lining cells of the cribriform component of the tumor. Present findings demonstrate that adenoid cystic carcinoma of the external auditory canal had dual epithelial and myoepithelial differentiation and can mimic the ceruminous glands of the auditory canal.

Actins↗
PubMed1993

A comparative study of vascular proliferation in brain metastasis of lung carcinomas.

Because of the marked vascular proliferation seen in brain metastases of small cell carcinoma of the lung (SCCL), we studied the morphometric and immunohistochemical characteristics of proliferating vessels in metastases from 20 autopsy cases of SCCL with brain metastasis. These were compared with those in surgically resected brain metastases of lung carcinomas, including 6 cases of SCCL, 19 cases of adenocarcinoma and 5 cases of squamous cell carcinoma. Angiogenesis in the tumours was scored by the microscopic angiogenesis grading system (MAGS). The MAGS score for autopsy and surgical metastatic lesions was highest in SCCL. Histologically, many vascular glomeruloid structures were formed in the brain metastases of SCCL, and immunohistochemistry revealed that these lesions were composed of proliferating endothelial cells and pericyte/smooth muscle cells. Immunostaining for basic fibroblast growth factor, a potent angiogenic factor, showed immunoreactivity in the tumour cells, regardless of histological type, and in the surrounding glial cells. Complex autocrine and paracrine phenomena participate in the development of metastatic cerebral lesions with vascular proliferation.

Adenocarcinoma↗
PubMed1992

Uptake and intracellular transport of cationic ferritin in the bronchiolar and alveolar epithelia of the rat.

Cationic ferritin was used as a marker to reveal the processes of endocytosis and intracellular transport in bronchiolar and alveolar epithelia. The marker was injected into the lung via the trachea, and ultrastructural observation of the distribution of ferritin particles in bronchiolar and alveolar epithelial cells was carried out at intervals of 5, 15, 30 and 60 min after the injection. The luminal surface of the airway and the alveolar epithelium showed diffuse labeling with cationic ferritin. In general, ferritin particles were observed in vesicles and vacuoles of the bronchiolar and alveolar epithelial cells within 5 min of injection; they appeared in multivesicular bodies within 15 min. Multivesicular bodies and secondary lysosomes containing ferritin particles, some of which showed a positive reaction for acid phosphatase, were seen in the basal cytoplasm within 30 min; ferritin particles appeared in the basal lamina below the Clara cells, ciliated cells and type 2 alveolar cells within 30 min. Ferritin particles were seen in ovoid granules of some Clara cells and in lamellar inclusion bodies of many type 2 alveolar cells. Brush cells and type 1 alveolar cells took up only a small quantity of ferritin particles.

Animals↗
PubMed1992

Development of hepatocellular carcinoma associated with increases in DNA synthesis in the surrounding cirrhosis.

The relationship between DNA synthesis activity in hepatocytes from cirrhotic tissue and development of hepatocellular carcinoma was studied in 33 posthepatitic patients with Child's grade A cirrhosis. DNA synthesis activity was measured by a bromodeoxyuridine (a thymidine analogue) labeling index, using the bromodeoxyuridine-antibromodeoxyuridine in vitro method, and the patients were followed up prospectively with frequent liver ultrasonography for 2 years. During the 2-year follow-up, 11 of the 33 cirrhotic patients developed hepatocellular carcinoma; they included 8 of the 15 patients (53%) in the high labeling index (greater than 1.5%) group compared with only 3 of the 18 patients (17%) in the low labeling index (less than 1.5) group (P less than 0.05). Five of the latter 18 subsequently had increased synthesis activity. Of these 20 patients who showed high synthesis activities either initially or subsequently, 10 (50%) developed hepatocellular carcinoma, in contrast to 1 of 13 (8%) with persistently low activities (P less than 0.05). Thus, hepatocellular carcinoma seems to develop or may become detectable when DNA synthesis in the background cirrhosis is increasing or remains high.

Adult↗
PubMed1992

Modulation of the incidence of hamster pulmonary endocrine cell hyperplasia by unilateral collapse of the lung.

We evaluated the effects of the unilateral collapse of the left lung on the formation of pulmonary endocrine cell hyperplasia (PECH) induced by 4-nitroquinoline 1-oxide (4NQO) in Syrian golden hamsters. Ten hamsters were injected subcutaneously with 20 mg/kg body weight of 4NQO, once a week for 4 weeks and treated with injection of silicone rubber into the left thorax at the 8th experimental week. Another 30 animals were divided into three groups: treated with 4NQO only, left lung collapse only, and vehicle only. All animals were sacrificed at the 30th week. The lung tissues were embedded in paraffin; 50 serial sections were made from the tissues of each animal and were studied histologically and immunohistochemically. PECH showed a positive immunostain for calcitonin and/or serotonin. In the uncollapsed right lungs of the animals treated with both 4NQO and unilateral collapse, the mean incidence of PECH was 12.2 x 10(-2)/mm3 lung volume; the incidences of PECH in the animals treated with 4NQO only, collapse only, and vehicle only were 4.1, 3.8, and 1.4 x 10(-2)/mm3, respectively. In the collapsed left lungs, PECH did not form, regardless of 4NQO treatment. This study demonstrates that unilateral collapse of the lung modulates the incidence of PECH induced by 4NQO in hamsters.

4-Nitroquinoline-1-oxide↗