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Biomedical subjects

S Nishikawa

Publications and source records attributed to S Nishikawa.

At least 199 records · Page 11Linked to original sources
PubMed1996

Involvement of Fyn tyrosine kinase in progression of cytokinesis of B lymphocyte progenitor.

We analyzed the role of Fyn tyrosine kinase in cell cycle progression of B lymphocyte progenitor (pro B cell). Whereas there were no substantial defects in the intramarrow B cell genesis in the fyn(-) mouse, and long-term proliferation of fyn(-) pro B cells was maintained in vitro under a serum containing culture condition, the cell cycle was arrested at G2/M upon serum deprivation. Morphological analyses demonstrated that the cytokinesis of fyn(-) pro B cells was retarded in the presence of serum and that the entry of fyn(-) pro B cells into late telophase was completely blocked under the serum-free condition. In contrast, the earlier phases of mitosis of fyn(-) pro B cells proceeded normally without FCS. This failure to initiate late telophase resulted in the accumulation of elliptical binucleated cells that might be the outcome of the nuclear division without cytokinesis. Consistent with this defect in the progression of cytokinesis, Fyn was localized in the midspace of dividing pro B cells at anaphase. These results suggested that Fyn localizes at the midspace of dividing pro B cells and regulates the progression of cytokinesis.

Animals↗
PubMed1996

Evaluation of routine sonography for early detection of pancreatic cancer.

The diagnostic accuracy of routine abdominal sonography for the detection of pancreatic cancer was examined. During the one-year period of 1994, sonographic examination of the upper abdominal region was performed 12,761 times on a total of 9410 patients for the screening of abdominal disorders. In 655 cases (7%) part of the pancreas could not be observed. Based on the "Diagnostic criteria for pancreatic cancer" published by the Japanese Society of Ultrasound in Medicine, sonographic finding was evaluated to be positive for pancreatic tumor in a total of 411 cases. At the end of 1995, 51 patients were proven to have pancreatic cancer, and 45 of these cases were ductal adenocarcinoma. In 26 cases the tumor was surgically resected. Fifty cases were true sonographic positives and one was a false negative. The sensitivity, specificity, overall accuracy, and positive and negative predictive values of sonography for pancreatic cancer were 98.0%, 95.9%, 95.9%, 12.2% and 100.0%, respectively. Among the 50 true positive cases, the tumor diameter was less than 1 cm in four (8%). In conclusion, the diagnostic accuracy of sonography for the detection of pancreatic cancer is sufficiently high. Therefore, a detailed study aimed at mass screening for pancreatic cancer using sonography as the main modality seems warranted as a countermeasure for the rapid increase of pancreatic cancer in Japan.

Abdomen↗
PubMed1996

Involvement of platelet-derived growth factor receptor-alpha in hair canal formation.

Hair follicles develop and are maintained by multiple rounds of inductive events involving interactions among various cell types within the follicles and the adjacent mesenchyme. Although evidence suggests that several growth factors, cell adhesion molecules, and transcriptional regulators are involved in those cell-cell interactions, the molecular mechanisms regulating each pivotal step of hair follicle development, such as formation of the hair germ, root sheath, sebaceous gland, and hair canal, remain largely unknown. In this study, we established the antagonistic monoclonal antibody APA5 against platelet-derived growth factor (PDGF) receptor-alpha (PDGFR-alpha) and used it to investigate the role of PDGFR-alpha in neonatal skin development. In addition to the dermal mesenchyme, a known site of PDGFR-alpha expression, immunohistologic staining of neonatal skin detected transient expression of PDGFR-alpha in the perinatal epidermis for several days. On the other hand, ligands for PDGFR-alpha were detected in epithelial cells and sebaceous glands of hair follicles. To determine whether this contiguous expression of PDGF and PDGFR-alpha in neonatal skin plays a functional role, we injected APA5 into neonates to block the function of PDGFR-alpha. Consistent with the PDGF/PDGFR-alpha expression in the neonatal skin, two defects were induced by this procedure. First, hair canal formation in the epidermis was severely suppressed. Second, the growth of dermal connective tissues and of hair follicles of pelage hairs was suppressed. These results indicate that PDGF signals are involved in both the epidermis-follicle interaction and the dermal mesenchyme-follicle interaction required for hair canal formation and the growth of the dermal mesenchyme, respectively.

3T3 Cells↗
PubMed1996

Endogenous interleukin-4, but not interleukin-10, is involved in suppression of host resistance against Listeria monocytogenes infection in interferon-depleted mice.

The production and roles of endogenous interleukin-4 (IL-4) and IL-10 in a sublethal infection with Listeria monocytogenes were studies in normal mice and anti-gamma interferon (IFN-gamma) monoclonal antibody (MAb)-pretreated mice. In normal mice, the expression of mRNAs for IL-4 and IL-10, which was amplified by reverse transcription-PCR, was induced in the spleens and livers either early or late in infection, although the serum IL-4 and IL-10 were not detectable by enzyme-linked immunosorbent assays. In vivo administration of anti-IL-4 MAb showed no effect on antilisterial resistance, whereas anti-IL-10 MAb partially diminished the defense. In anti-IFN-gamma MAb-pretreated mice, a delay in the bacterial elimination from the spleens and livers was observed and high titers of serum IL-4 and IL-10 were induced late in infection. Production of endogenous IL-4 and IL-10 was suppressed in both CD4+ cell-and CD8+ cell depleted mice. The suppression of antilisterial resistance in anti-IFN-gamma MAb-pretreated mice was canceled when anti-IL-4 MAb was injected late in infection, whereas anti-IL-10 MAb showed no effect. These results suggest that the host immune responses were polarized into the T-helper 2 phenotype in anti-IFN-gamma MAb-pretreated mice and that inhibition of host resistance against L. monocytogenes by depletion of IFN-gamma might be attributable to IL-4 produced by T cells polarized into the T-helper 2 phenotype as well as the inhibition of the IFN-gamma effects.

Animals↗
PubMed1996

Transforming growth factor beta is protective in host resistance against Listeria monocytogenes infection in mice.

The role of transforming growth factor beta (TGF-beta) in host resistance against Listeria monocytogenes infection was studied with mice. The constitutive expression of TGF-beta 1 mRNA was observed in the spleens and livers of mice before and after infection. Injecting the mice with anti-TGF-beta 1 peptide serum resulted in diminished antilisterial resistance, whereas the administration of human platelet-derived TGF-beta 1 enhanced the resistance. Moreover, mice were protected against lethal infection when treated with TGF-beta 1. These results suggest the TGF-beta 1 might be involved in antilisterial resistance. On the other hand, injecting the mice with TGF-beta 1 resulted in a decrease in the titers of endogenous gamma interferon, tumor necrosis factor alpha, and interleukin-6, which are crucial in antilisterial resistance, in sera and in extracts of spleen and liver. Thus, a complicated mechanism might be involved in the role of TGF-beta 1 in host resistance against L. monocytogenes infection.

Animals↗
PubMed1996

Impairment of Kit-dependent development of interstitial cells alters contractile responses of murine intestinal tract.

We examined developmental changes in responses of the isolated segment of the ileum of BALB/c mice treated with a monoclonal antibody (ACK2) to the receptor tyrosine kinase (Kit) for 4 days postnatally to pharmacological agents in vitro. Rhythmic contraction and relaxation of the isolated ileum started to appear on day 4 postpartum, and the sensitivity to acetylcholine (ACh) decreased gradually after birth. Treatment with ACK2 induced augmentation of contractile responses and receptor sensitivity of the longitudinal muscle of the ileum to ACh, bradykinin, and prostaglandin F2 alpha. ACh induced larger depolarization in smooth muscle cells of the ileum in the ACK2-treated mice than in the control. Circular muscle responses to these substances, as measured by changes in intraluminal pressure, were not altered by ACK2 treatment. Results suggest that interstitial cells play an important role not only in the development of the pacemaking system of the small intestine but also in the functional development of the contractile properties of the intestinal smooth muscle.

Acetylcholine↗
PubMed1996

Phagocytotic processing of apoptotic bodies of transitional ameloblasts by MHC Class II-expressing macrophages in rat incisor.

We examined the processing of apoptotic ameloblast fragments at the transitional stage by immunocytochemistry and electron microscopy. Both macrophages and epithelial cells in the enamel organ ingested apoptotic bodies. MHC Class II antigen expression and the phagolysosomal nature of the macrophages were further examined by immunoelectron microscopy using OX6 and ED1 antibodies followed by HRP- or gold-conjugated secondary antibodies. The results showed that macrophages in the enamel organ at the transitional stage expressed MHC Class II antigens at their plasma membranes and that the same cells contained apoptotic fragments in ED1-positive vacuoles. We suggest that diverse immunological responses, such as the self-antigen presentation, may be invoked in the process of apoptosis.

Acid Phosphatase↗
PubMed1996

Internalization of styryl dye FM1-43 in the hair cells of lateral line organs in Xenopus larvae.

We used a fluorescent dye, FM1-43 to investigate mechanotransduction mechanisms in the hair cells of lateral line organs of Xenopus larvae. FM1-43 specifically labeled the hair cells. The photo-oxidation technique was performed with election microscopy to examine the labeling sites and their mechanisms. The results showed that the labeling sites were mitochondria and rough endoplasmic reticulum throughout the cytoplasm. Endocytic activity of the hair cells was limited to endosomes and small granules located at the apical part of the cells. Blockers of the mechanosensitive cation channel (neomycin, gentamicin, streptomycin, and amiloride) effectively inhibited FM1-43 labeling. One of the blockers, amiloride, was found to bind to hair cells when its fluorescence was examined. Divalent cations such as Mg2+ and Ca2+, but not monovalent cations such as Na+ and K+, inhibited FM1-43 labeling when they were added in excess amounts. These results suggest that FM1-43 internalizes hair cells via the putative mechanosensitive cation channel in the plasma membrane.

Amiloride↗
PubMed1996

Distinct stages of melanocyte differentiation revealed by anlaysis of nonuniform pigmentation patterns.

The injection of an antagonistic anti-murine c-kit monoclonal antibody ACK2 during mouse embryonic development produced three distinctive pigmentation patterns on the coat of the offspring. Pattern 1 consisted of pigmentation in craniofacial and caudal regions and was induced by an ACK2 injection between 9.5 and 11.5 days post coitum (dpc). In pattern 2, the entire coat was unpigmented and was induced by the injection at around 13.0 dpc. Pattern 3 consisted of pigmented patches spreading ventrolaterally from the dorsoanterior trunk regions towards the anterior and posterior directions and it was induced by ACK2 administered at 14.5-15.0 dpc. We investigated the embryological basis of these nonuniform pigmentation patterns to elucidate the process of melanoblast differentiation between lineage commitment and colonization into developing hair follicles. The results showed the following. (1) Melanocyte differentiation at the embryonic stage from 10.5 to 12.5 dpc progresses in a spatially nonuniform fashion, being faster in the craniofacial and caudal regions than in the trunk; pattern 1 reflects this. (2) Melanoblasts are activated to proliferate synchronously upon entering into the epidermis; pattern 2 correlates with this process. (3) c-kit functions as a survival signal for proliferating melanoblasts in the epidermis. (4) The melanoblasts that enter developing hair follicles can survive without a c-kit signal; pattern 3 essentially represents the hair follicles colonized by these cells. Analysis of the melanoblast distribution of ls/ls embryos that bear a loss-of-function mutation in the endothelin 3 gene suggested that endothelin 3 is required for early melanoblast differentiation before entering into the epidermis, whereas proliferation in the epidermis takes place without this molecule. Based on these data, we propose 4 distinct steps of embryonic melanocyte differentiation: (1) migration in the dermis, which requires both c-kit and endothelin 3; (2) a state before epidermal entry that is resistant to anti-c-kit mAb; (3) cell proliferation after entering the epidermal layer, which requires c-kit and endothelin receptor B but not endothelin 3 and (4) integration into developing hair follicles, which renders melanoblasts resistant to anti-c-kit mAb. Thus, melanoblast differentiation proceeds by alternately repeating c-kit -dependent and c-kit-independent stages and c-kit functions as a survival factor for the proliferating melanoblasts.

Animals↗
PubMed1996

Effects of ganglioside GD1a on the fluidity and phase transition of phosphatidylcholine model membrane immobilized on porous cellulose acetate filter.

The thermotropic effect of ganglioside GD1a on the phosphatidylcholine membrane immobilized on the porous cellulose acetate filter was investigated by ESR spin-labeling and DSC. The spin-labeled GD1a having 12-DOXYL-stearic acid instead of the long acyl chain of the ceramide portion (GD1a*) was incorporated into the model membrane. An ESR examination of the membrane showed that GD1a* undergoes an anisotropic rotation in the wide range of temperature -30-70 degrees C. By monitoring the overall splitting (2T parallel) of spin-labeled phosphatidylcholine (PC*), the model membranes were found to show decreased fluidity in accordance with the GD1a content. The phase transition temperature (Tm) of distealoylphosphatidylcholine (DSPC) model membranes could be estimated by the measurement of ESR and DSC. The effects of GD1a were found to be more significant in broadening the phase transition rather than in elevating the Tm of DSPC model membranes.

Animals↗
PubMed1996

Pathogenesis of drug-induced gingival overgrowth. A review of studies in the rat model.

Drug-induced gingival overgrowth is a side effect associated principally with 3 types of drugs: anticonvulsant (phenytoin), immunosuppressant (cyclosporine A), and various calcium channel blockers (nifedipine, verapamil, diltiazem). In this review, we describe the features of phenytoin-, cyclosporine A- and nifedipine-induced gingival overgrowth in rats and discuss factors influencing the onset and severity of these disorders. There are several features common to the gingival overgrowth induced by these drugs: 1) gingival overgrowth is more conspicuous in the buccal than in the lingual gingiva and less severe in the maxilla than in the mandible; 2) once the blood concentration of the drug reaches a certain level as a result of increasing the dose, the incidence of gingival overgrowth is 100% and its severity is dependent on the blood level, the most severe overgrowth being induced by cyclosporine A; 3) the duration of drug administration for maximal gingival overgrowth to develop is about 40 days; 4) the gingival overgrowth regresses spontaneously after discontinuing the drug; 5) accumulation of dental plaque is not essential for the onset of overgrowth, but plays a role in its severity; and 6) more severe overgrowth is induced in young than in old rats. Furthermore, male rats are more susceptible than females to nifedipine-induced gingival overgrowth. These results suggest that drug-induced gingival overgrowth in rats is dependent on the oral drug dose, blood drug level, age, and sex and that preexisting gingival inflammation is a factor relevant to its severity. Since these factors have also been suggested to be important determinants for human drug-induced gingival overgrowth, the rat model may prove valuable in the future for elucidating the molecular pathogenesis of the disorder.

Age Factors↗
PubMed1996

Ultrastructure of hyaluronic acid and collagen in the human vitreous.

The ultrastructure of hyaluronic acid (HA) and collagen in the human vitreous was investigated. After fixation with 4% glutaraldehyde and 0.1% cetylpyridinium chloride (CPC), sectioned blocks of retina and attached vitreous body were stained with 500 ppm ruthenium red (RR) in 1.25% glutaraldehyde, postfixed with 500 ppm RR in 2% OsO4, and embedded in Epon 812. In addition, collagen II immunostaining was done. Collagen fibrils were coated with amorphous material believed to be HA. No amorphous material was seen in the fibrils in the cortical areas adjacent to the retina, but the amount of material gradually increased toward the center of the vitreous body. By using anticollagen II antibody, we showed the presence of collagen II, often oriented perpendicular to the surface. The central vitreous demonstrated some dense materials that were collagen fibrils packed into bundles of parallel fibrils. In summary, collagen fibrils contained little HA in the cortical areas adjacent to the retina, but they were packed into bundles of parallel fibrils in the central vitreous.

Collagen↗
PubMed1996

Localization of proline-like immunoreactivity in young rat retinal neurons.

PURPOSE: To localize proline in the rat retina. METHOD: We prepared antibodies raised against proline coupled to bovine serum albumin (BSA) with glutaraldehyde. To confirm the specificity of the antiserum, crossreactivity with other amino acids was determined using an immunodot procedure. Rat eyes were enucleated on postnatal day 21. Immunohistochemistry was performed on semi-thin sections using gold-labelled secondary antibodies and a silver-enhancement technique. RESULTS: Immunodots revealed that the antiserum was specific for proline. Amino acids from retinal extracts were separated on gel, transferred to BSA-glutaraldehyde treated filters, and stained with antibodies to determine if our antibody reacted only with proline. Staining was most intense on postnatal day 21 and disappeared in the adult retina. In addition to Muller cells, horizontal cells, amacrine cells, and some cells in the ganglion cell layer were labelled. CONCLUSIONS: Antibodies raised against proline revealed a transient immunoreactivity in young rat neurons. Our findings strongly indicate that the retina may express a high level of proline in the developmental stages. This result may provide a clue for clarifying the role of proline in the retina.

Animals↗
PubMed1996

Light-evoked changes of glycine-like immunoreactivity in the rat retina.

Glycine is an important inhibitory neurotransmitter in the vertebrate retina. Although many reports of glycine localization by the immunocytochemical method have been published, none has distinguished the neurotransmitter function of glycine from metabolic change. If glycine is the neurotransmitter, it seems that it must be affected or released by stimulation. We evaluated the possibility that glycine in retinal synapses changes with light stimulation. Glycine-like immunoreactivity was found by light microscopy to be present mainly in the inner plexiform layer and amacrine cells. In the inner plexiform layer these immunoreactive products decreased after illumination. The distribution of glycine-like immunoreactivity in the rat retina was also investigated after embedding anti-rabbit IgG-gold colloidal particles (GCP). In the dark-adapted eyes, glycine-like immunoreactivity was marked in the inner plexiform layer (27.8 +/- 5.7 GCP/microns2) and amacrine cells (33.5 +/- 5.5 GCP/microns2). This immunoreactive product significantly decreased only in the inner plexiform layer after illumination (15.5 +/- 4.5 GCP/microns2). These changes in immunoreactivity may reflect the release of endogenous glycine. Our results provide evidence for the role of glycine as a retinal neurotransmitter.

Animals↗
PubMed1995

Functional hierarchy of c-kit and c-fms in intramarrow production of CFU-M.

Whereas the molecular natures of M-CSF/CSF-1 and its receptor c-fms are well characterized, its actual role in the intramarrow hematopoiesis remains obscure. This is because disruption of this signaling pathway results in the osteopetrosis mouse that lacks the bone cavity for hematopoiesis. To elucidate the role of c-fms in intramarrow hematopoiesis, we produced an antagonistic monoclonal antibody to murine c-fms and investigated its expression and function in the normal bone marrow. c-fms+ cells were detected both in mature and immature hematopoietic cells. Morphologically, c-kit+c-fms-, c-kit+c-fms+ and c-kit-c-fms+ cells were medium sized blasts, large promyelocytes with azurophilic granules and mature monocytes respectively. CFU-M was 10-fold more enriched in the c-kit+c-fms- than c-kit+c-fms+ fraction. Moreover, injection of the anti c-fms antibody had no effect on the production of CFU-M in the bone marrow, while anti-c-kit mAb could deplete them. As c-kit+c-fms+ cells were readily generated in the culture of c-kit+c-fms- cells, most of the CFU-M in the bone marrow are, in fact, c-fms- cells that differentiate into c-fms+ upon culture. These observations indicate a clear functional hierarchy of c-kit and c-fms in the bone marrow. Namely, c-kit plays the primary role in the production and maintenance of CFU-M, while c-fms, though it co-expressed with c-kit and functions as the growth receptor for M-CSF in the culture, has only a minimum role in the proliferation of c-fms+ cells in the bone marrow.

Animals↗
PubMed1995

Identification of the control regions for mouse c-kit gene transcription induced by retinoic acid.

The c-kit gene encodes a receptor tyrosine kinase that reveals the pleiotropic effects both in the developing embryo and in the adult animal. In this study, we characterized transcriptional control of the c-kit gene. F9 teratocarcinoma cells differentiated and expressed the c-kit gene upon exposure to retinoic acid (RA). We isolated c-kit genomic DNA from approximately 20 kb upstream to approximately 10 kb downstream of the transcribed region. We identified two control regions for c-kit gene transcription by fusing genomic sequences to bacterial chloramphenicol acetyl transferase reporter genes and introducing the resultant constructs into differentiated F9 cells. The fragments, Sac I-Apa I in the 9th intron and Xho I-Kpn I in the 13th intron induced reporter gene activities. Nucleotide sequencing of the regions showed in that there were no consensus sequences of the RA receptor or cyclic AMP-responsive elements. Cyclic AMP augmented the expression of the c-kit gene induced by RA; when combined with RA, it was less effective on the reporter gene activity. In contrast to other reports, continuous exposure to the reagents was not required for c-kit expression. These findings suggested that transcription of the c-kit gene is regulated by complex controls and that there are different control regions responsive to RA and cyclic AMP.

Animals↗
PubMed1995

Overexpression of retinoic acid receptor alpha suppresses myeloid cell differentiation at the promyelocyte stage.

Retinoic acid receptor (RAR) alpha is required to heterodimerize with retinoid X receptor (RXRs) in order to regulate myeloid differentiation. If so, it is expected that overexpression of normal RAR alpha may perturb the RAR alpha/RXR heterodimer formation and also the differentiation of myeloid cells. We have described here the morphology and the RA response of human RAR alpha cDNA transduced murine bone marrow cells using a retroviral vector. Most of RAR alpha transduced cells displayed promyelocyte like morphology and their proportion of c-kit expressing population was increased remarkably compared with the control (Neor gene transduced cells). Furthermore, this morphology was observed even after these cells were brought into the semisolid culture containing IL-3 alone. Interestingly, immature RAR alpha transduced cells differentiated into mature granulocytes under the condition of the high concentration of RA(10(-6) M). We did not observe any effect of RAR alpha on monocytes. These results indicate that overexpression of normal RAR alpha is sufficient for inducing maturation arrest of myeloid cell lineage that is similar to the phenotype found in the acute promyelocytic leukemia bearing PML-RAR alpha translocation.

3T3 Cells↗