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K Uryu

Publications and source records attributed to K Uryu.

36 records · Page 2Linked to original sources
PubMed1996

Expression of tropomyosin isoforms in benign and malignant human breast lesions.

High molecular weight tropomyosins (tms) are commonly down-regulated in fibroblasts transformed by oncogenes. Previous studies have also demonstrated that specific tm isoforms are down-regulated in human breast carcinoma cell lines. We examined tropomyosin isoforms in cells prepared from non-cancerous breast lesions and primary human breast carcinomas. The average level of expression of all three high molecular weight tm isoforms (tm 1-3) in carcinomas was generally found to be less than 25% of that observed in non-cancerous breast lesions. Interestingly, the expression of tm 1 was found to be 1.7-fold higher in primary tumours with metastatic spread to axillary lymph nodes compared with primary tumours with no evidence of metastasis (p<0.05). Similarly, tm 1 expression was higher in two 12V-H-ras transformed fibroblast cell lines capable of experimental metastasis compared with three weakly metastatic cell lines. We conclude from these studies that expression of high molecular weight tm isoforms is low in primary breast carcinomas, and that metastatic tumours express relatively high levels of tm 1.

Breast↗
PubMed1995

Sample preparation of human tumors prior to two-dimensional electrophoresis of proteins.

Sample preparation procedures are critical for obtaining high quality two-dimensional electrophoresis (2-DE) separations from clinical tumor materials. In this communication, we describe procedures which we have found satisfactory for preparing various tumor samples for 2-DE. By these procedures, tumor cells are enriched from serum proteins and contaminating stromal cells. Tumors can be prepared with good reproducibility to obtain high resolution using these techniques.

Electrophoresis, Gel, Two-Dimensional↗
PubMed1995

Immunocytochemical identification of serotonergic supraependymal nerve fibers in the third ventricle of the house musk shrew, Suncus murinus.

Supraependymal fibers of the house musk shrew (Suncus murinus) were examined by conventional scanning electron microscopy (SEM), backscattered electron (BSE) imaging of enzyme immunohistochemistry and by immunotransmission electron microscopy in the dorsal part of the third ventricular wall. In this region, ependymal cells were not so heavily ciliated and conventional SEM studies showed two main categories of supraependymal fibers. The first type consisted of long fibers fasciculated which were distributed over the ventricular surface between the anterior commissure and the subfornical organ. The second category was a thin fiber which was observed on the ependymal luminal surface. Some of these fibers had varicoses or terminal-like swellings. This type of supraependymal fiber seemed to originate in the first type of fiber bundles. To confirm the nature and the distribution of serotonin-immunoreactive supraependymal fibers, BSE imaging using immunohistochemical reactions was used. Serotonin-immunoreactive structures were shown as highlighted structures by means of a backscattered electron mode. These investigations revealed that the majority of both types of supraependymal fibers observed by conventional SEM contained serotonin. A moderate number of serotonergic supraependymal fibers was observed on the ventricular surface of the subfornical organ. Immunohistochemical studies using Vibratome sections of identical ventricular regions revealed the presence of serotonin-immunoreactive processes, with the use of light- and electron-microscopy. They were distributed in in the third ventricle just adjacent to the ependymal luminal surfaces. These fibers contained immunoreactive large cored vesicles and immunonegative small clear vesicles.

Animals↗
PubMed1995

Detection of polypeptides associated with the histopathological differentiation of primary lung carcinoma.

Two-dimensional polyacrylamide gel electrophoresis combined with a non-enzymatic sample preparation technique is useful for analysing clinical tumour material. Using these techniques, we analysed the relationship between the histopathological findings in primary lung malignancies and the expression of a number of unidentified polypeptides that were detected in the molecular weight region 20-35 kDa. In this study 45 cases of primary lung cancer (PLC) (21 cases of adenocarcinoma, ten cases of squamous cell carcinoma, five cases of large-cell carcinoma, one case of adenosquamous cell carcinoma, five cases of small-cell carcinoma and three cases of carcinoid tumour) were examined. For reference, a human diploid fibroblast cell line (W138) and normal peripheral lymphocytes were used. Sixteen polypeptides were judged to be associated with histopathological features. These polypeptides seem to be valuable as differentiation markers. The simultaneous evaluation of these polypeptides and some other proliferation markers (e.g. PCNA, PCNA 'satellite', Numatin/protein B23 and lamin B) seems to clarify the characteristics of each case of PLC. Furthermore, it is possible to classify PLC based on the two-dimensional electrophoresis findings, and this classification of PLC is suggested to reflect the biological features of the tumour more precisely than that based only on morphology.

Adenocarcinoma↗
PubMed1995

[CT measurement of the thoracic density].

The purpose of this introducing study was to establish the normal lung density using computed tomography of the chest. If it is possible to analyze the density of normal lung parenchyma, it could be possible to estimate the quantity of effective ventilation capacity based on account of the CT number of given range. A cross section of the lung, however, is not homogeneous because it contents of vascular tissue, interstitial tissue, air and others which have widely variety densities. To estimate pulmonary function, it is necessary to know the exact CT number of only normal lung tissue. Mean values for varies tissues and areas in the thorax were assessed in 34 patients (21 males, 13 females, average age: 58 years, from August 1987 to June 1993), 18 case of lung cancer (10 preoperative, 8 postoperative), 5 metastatic lung cancer, 1 giant bulla, 1 aspergilloma, 1 pulmonary AV fistula, 8 case had no lung disease. The measured normal peripheral lung by radiographic appearance on CT images, vascular in the lung, bronchus, rib, muscle and fat structured chest wall, heart, liver, aorta, esophagus, and thymus. Density of normal peripheral lung tissue was measured in 6 points on each slices. These scores were compared left vs. right, slice number, and anterior vs. posterior. The CT numbers in this study following the no enhanced computed tomography scanner: GE9800 CT by using the regions of interest which limited in 0.01 cm 2 area. There was no significant difference between left and right lung, among different slices, although between anterior and posterior in the normal lung.(ABSTRACT TRUNCATED AT 250 WORDS)

Female↗
PubMed1993

An immunohistochemical study of basic fibroblast growth factor in the developing chick.

An antiserum against basic fibroblast growth factor (bFGF) was characterized by immunoblot experiments and used to investigate immunohistochemically the appearance of bFGF-like immunoreactivity in the developing chick. Crude homogenates of chick embryos at every developmental stage, when subjected to immunoblotting with the use of bFGF antiserum, exhibited a main band with the same molecular weight (18 kDa) as bovine bFGF. With immunohistochemistry, bFGF immunoreactivity (bFGF-IR) was detected exclusively in intracellular components of various tissues at different stages of development; bFGF-IR appeared initially on embryonic (incubation) day 3 (E3) in the myotome, on E12 in the spinal cord and ganglia, on E8 in chondrocytes and osteoblasts of the vertebrae, and on E10 in the esophageal epithelium. Immunoreaction products were present either in the cytoplasm or in the nuclei, depending on the types of individual bFGF-containing cells; developing chondrocytes and cells in the stratum basale of the esophagus exhibited intense immunoreactions exclusively within the nuclei, and the other cells mainly within the cytoplasm. Moreover, bFGF-IR was observed in discrete regions of these tissues at different stages; the epithelium of the esophagus contained bFGF-IR in all layers on E10 to E18 with a superficial-to-basal gradient, but it began to exhibit bFGF-IR only in the stratum basale after E20; and bFGF-IR was more abundant in hypertrophic chondrocytes than in proliferating ones. As chicks aged, bFGF-IR decreased or disappeared in the muscles, vertebrae and esophageal epithelium, but neuronal bFGF increased in intensity until the perinatal period and thereafter remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Basic fibroblast growth factor-like immunoreactivity in the rat trigeminal sensory system and peri-oral skin with vibrissae.

We have characterized an antiserum against basic fibroblast growth factor (bFGF) by immunoblot, investigated the location of bFGF-like immunoreactivity (bFGF-IR) in the trigeminal sensory system and perioral skin endowed with vibrissae, and demonstrated the site of bFGF mRNA expression in the vibrissae by in situ hybridization histochemistry. Light-microscopic immunohistochemistry has demonstrated that bFGF-IR is present not only in trigeminal ganglion neurons and their central and peripheral processes, but also in cells of the matrix, external root sheath and papillae of vibrissae and the stratum basale of the stratified squamous epithelium of the skin. Electron microscopy has revealed intense bFGF-IR mainly in cytoplasmic regions, other than the lumen of rough endoplasmic reticulum and the Golgi apparatus, in trigeminal ganglion neurons, in fibroblast-like cells in the papillae, and in capsules of vibrissae. In contrast, actively proliferating and/or differentiating cells in the matrix of vibrissae have intensely stained euchromatin and weakly labeled cytoplasm that, unlike that of the aforementioned cells, contain immunoreaction products in discrete spots less than 100 nm in diameter, implying the generation of different molecular forms of bFGF in cells of the matrix and papillae. Moreover, the accumulation of bFGF in the euchromatin appears to take place in cells at non-mitotic stages (possibly interphases), characterized by a conspicuous nucleolus and well-developed nuclear envelope. A digoxigenin-labeled cRNA probe for the demonstration of bFGF mRNA gives conspicuous hybridization signals mainly in the matrix of vibrissae. These findings suggest that bFGF is involved in the growth and differentiation of matrix cells during certain periods of the cell cycle and that it acts as a non-mitogenic mediator in the adult trigeminal sensory system.

Animals↗
PubMed1993

Glycoconjugate histochemistry of the subcommissural organ in the domestic chicken: lectin histochemistry using the avidin-biotin-peroxidase complex (ABC) method.

The properties and distribution of the secretory substance in the ependymal cells of the subcommissural organ in the domestic chicken were examined with seven different lectins, using the avidin-biotin-peroxidase complex method. On the ventricular surface of this organ, LFA, PNA, RCA-I and WGA reacted positively. In the apical cytoplasm, Con A, LFA, RCA-I and WGA gave positive reactions. On the other hand, only Con A and WGA gave positive reactions in the basal cytoplasm. In the hypendymal vascular wall, LFA, RCA-I and WGA reacted positively. Neuraminidase digestion completely inhibited reactions with LFA and reduced the WGA reaction within the apical cytoplasm. At the same time, this digestion disclosed a moderately positive reaction with PNA in the apical cytoplasm. The significant findings of this study may be summarized as follows: 1) the ependymal cells of the subcommissural organ in the domestic chicken synthesize a complex oligosaccharide for a secretory product which is discharged into the ventricular cavity; 2) in the basal cytoplasm and basal processes, only oligosaccharides containing large amount of mannose are present.

Animals↗
PubMed1993

Localization of basic fibroblast growth factor (bFGF)-like immunoreactivity in neural circuits innervating the gastrocnemius muscle, with reference to the direction of bFGF transport.

An antiserum against basic fibroblast growth factor (bFGF), characterized by immunoblot, was firstly used to immunolocalize bFGF-like materials in dorsal root ganglion (DRG) neurons, spinal motoneurons and their processes innervating the gastrocnemius muscle, and then secondly to determine the direction of bFGF transport in the processes by nerve ligation or nerve section experiments. Light and electron-microscopic immunohistochemistry revealed that significant numbers of DRG neurons (mainly of the large type), spinal motoneurons and their processes supplying the muscle are immunoreactive for bFGF, and that the bFGF neurons have diffuse immunoreaction deposits over the entire cytoplasmic areas except for the Golgi complex, mitochondria and microfilaments. Nerve terminals and glial processes with bFGF were rarely seen in contact with or in close apposition to the immunopositive neurons. At sixteen hours, 3 days and 7 days after unilateral ligation or transection of the sciatic nerve, bFGF fibers accumulated in a time-dependent way in the proximal but not in the distal stump; nerve transection caused a more intense accumulation of bFGF-immunoreactive fibers than did nerve ligation. These findings suggest that bFGF in large DRG neurons and motoneurons is transported anterogradely into the peripheral tissue, possibly without being incorporated into secretory granules or synaptic vesicles. They raise the possibility that bFGF within the transected nerve fibers is more actively conveyed during regeneration than in the ligated nerve fibers.

Animals↗
PubMed1993

Light- and electron-microscopic localization of basic fibroblast growth factor in adult rat retina.

An antiserum against basic fibroblast growth factor (bFGF) was characterized by immunoblot and used to localize bFGF-immunoreactivity in the adult rat retina. Crude homogenate of the retina, when examined with immunoblotting, exhibited a main band with the same molecular weight (18kDa) as bFGF. Light microscopic immunohistochemistry demonstrated intense bFGF staining in the photoreceptor outer segments and the outer plexiform layer, and weak immunostaining in some cells of the outer nuclear layer, inner nuclear layer and ganglion cell layer. The most intense bFGF-immunoreactivity was noted in glial (Müller) cells with vertically oriented thin processes. There were also immunoreactive pigment epithelial cells and mesenchymal cells in the choroid. Under electron microscopy, the outer plexiform layer was found to contain immunoreactive products exclusively in the axon terminals of photoreceptor cells which formed synapses with bipolar cell processes. The more widespread and uneven localization of bFGF-immunoreactivity in the mature rat retina than previously demonstrated suggests that endogenous bFGF is involved in complex physiological events other than retinal regeneration and histogenesis.

Animals↗
PubMed1992

Fine structure and possible origins of nerve fibers with corticotropin-releasing factor-like immunoreactivity in the rat central amygdaloid nucleus.

The fine structure of nerve fibers with corticotropin-releasing factor (CRF)-like immunoreactivity in the central amygdaloid nucleus and CRF-containing afferents to the nucleus were investigated by pre-embedding immunoelectron microscopy and by the combination of fluoro-gold tracing and the indirect immunofluorescence method. Significant numbers of CRF nerve endings and dendrites formed synapses with non-immunoreactive dendrites and axon terminals, respectively. Axon terminals devoid of CRF frequently made synapses with the soma of immunoreactive and non-immunoreactive neurons; CRF nerve endings in contact with the soma were fewer in number. Occasionally, CRF was localized to both pre- and postsynaptic structures in the central amygdaloid nucleus. After fluoro-gold injection into the central amygdaloid nucleus and adjacent areas, double-labeled cells with the tracer and CRF were observed mainly in the lateral hypothalamic area and occasionally in the dorsal raphe nucleus, and they were less numerous than single-labeled cells. These findings suggest that part of the CRF axon terminals identified in the electron micrographs arises from neurons in the lateral hypothalamic area and the dorsal raphe nucleus and the others from intra-amygdaloid CRF neurons. The immunoreactive dendrites are likely to derive from neurons in the central amygdaloid nucleus, which are shown to send axons to the lower brainstem. Thus, this study demonstrates that CRF structures constitute a more complex neuronal network in the central amygdaloid nucleus than previously considered.

Amygdala↗
PubMed1991

Development of tyrosine hydroxylase-like immunoreactive structures in the chick retina: three-dimensional analysis.

This study was designed to investigate the developmental profile of tyrosine hydroxylase-like immunoreactive structures in the chick retina in both frozen sections and wholemount preparations. In frozen sections, cells with tyrosine hydroxylase-like immunoreactivity were first detected in 10 to 15 cell rows from the innermost part of the inner nuclear layer on embryonic or incubation day 11. They were seen in the inner cell rows of the inner nuclear layer during later periods; by embryonic day 18, the immunoreactive cells were located 1 to 3 cell rows outward from the innermost part of the inner nuclear layer where mature immunoreactive cells mainly exist. The immunoreactive cells began to give rise to processes on embryonic day 13. The processes (possibly dendrites) gradually increased in number and intensity in sublayers 1 and 4 of the inner plexiform layer during prenatal life. Several days after hatching, an abrupt increase in immunoreactive processes was noted in sublayer 1 but not in sublayer 4. On the sixth postnatal day, retinal neural elements immunoreactive for tyrosine hydroxylase seemed to exhibit a distribution pattern similar to that of the adult chick. In wholemount retinas, immunoreactive cells were initially detected at the earliest stage of embryonic day 12 in a small circle termed "starting area" occupying the ventral part of the temporal retinal field. The closer to the "starting area," the earlier the retinal area began to express many immunoreactive cells. Thus tyrosine hydroxylase cell density in individual retinal areas, as represented by cell number per square millimeter, peaked in different developmental periods varying from embryonic day 12 to day 14. At this stage, immunoreactive cells were arranged irregularly in the retina. Thereafter, the cell density as well as total cell number gradually declined and reached a plateau around embryonic day 20 when tyrosine hydroxylase-like immunoreactive cells, like those in the mature retina, showed an even distribution throughout the retina.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1991

Differential lectin binding on walls of thoraco-cervical blood vessels and lymphatics in rats.

Lectin binding in the walls of large to medium-sized blood vessels and lymphatics in the rat thoraco-cervical region was examined histochemically. The tunica intima of the aorta and superficial cervical artery showed positive reactions with wheat germ agglutinin (WGA) and Concanavalin A (ConA) but not with Dolichus biflorus agglutinin (DBA). The tunica media of the aorta exhibited intense WGA binding, especially on the smooth muscle cells, but the tunica media of the superficial cervical artery did not react with the lectin. Neither ConA nor DBA bound to the tunica media of the aorta and superficial cervical artery. The tunica adventitia of both arteries contained sites binding the three lectins, although DBA reactivity declined as the vascular diameter decreased. The tunica intima of the superior vena cava and azygos vein exhibited positive WGA and ConA binding, whereas DBA binding was noted on only part of the tunica intima of the superior vena cava and not on that of the azygos vein. The tunica media and tunica adventitia were reactive for all three lectins. The WGA and ConA binding sites in the tunica adventitia showed loose networks, suggesting lectin binding on connective tissue elements interlacing among smooth muscle bundles. Lectin binding sites in the walls of lymphatics exhibited an arrangement similar to those in the walls of the veins. Moreover valves protruding into the lumen showed intense WGA and ConA binding and scattered DBA binding. Three other lectins (Ulex europaeus agglutinin, peanut agglutinin, Maclura pomifera) were examined, but they showed no reactions with the vessels. Thus, the differential binding of lectins on the walls of blood vessels and lymphatics of various sizes suggests the functional complexity of monosaccharide residues in the vascular walls.

Animals↗
PubMed1990

Ultrastructural and cytochemical studies of the subcommissural organ in the domestic chicken with special reference to the apical secretion.

Ultrastructural and cytochemical studies were undertaken with special reference to the apical secretion in the ependymal cells of the subcommissural organ in the domestic chicken. Glucose-6-phosphatase (G6Pase) and acid phosphatase (AcPae) were used as cytochemical markers for the endoplasmic reticulum (ER) and lysosomes, respectively. Periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) cytochemistry was employed to detect glycoconjugates. Most dense bodies were negative with AcPase staining. Dilated cisternal of ER were reacted with G6Pase cytochemistry. Golgi saccules and several kinds of spherical bodies were stained with PA-TCH-SP cytochemistry. Our results suggest that: 1) most dense bodies in the apical cytoplasm belong to the secretory granules discharged into the ventricular cavity, 2) dilated cisternae of the ER serve as the storage site of the substance which eventually becomes the secretory product, however, contents of the ER are not directly released into the ventricular cavity. Findings of this studies lead us to speculate that there are two intracellular transportation via different compartments: one is a transport via small vesicles from the perinuclear ER to the Golgi apparatus and the other is a transport bypassing the Golgi apparatus via cisternae of ER from the perinuclear ER to the supra-Golgi region. The substances in these two structures seem to be intermingled with each other and matured into secretory granules. This granules may be discharged into the ventricular cavity by exocytosis.

Acid Phosphatase↗
PubMed1989

Scanning electron microscopic studies on the embryonic development of the surface structures of the paraventricular organ in the Brown Leghorn chicken.

The development of the surface specialization of the paraventricular organ (PVO) was studied in the domestic chicken from the 10th embryonic day to the day of hatching by scanning electron microscopy. On the 10th embryonic day, the ventricular surface of the PVO was found to be covered with many oval-shaped processes. On the 12th embryonic day, an additional kind of elongated processes appeared in the dorsal area of the ventricular surface of the organ. From the 16th to 18th embryonic day, such elongated processes were present on the entire ventricular surface of the PVO. At the same stage, the elongated processes in the dorsal portion of the PVO began to form small, meshed networks over the surface of the ependyma. Both the oval-shaped processes and the elongated processes are thought to be dendritic terminals of the PVO neurons.

Animals↗
PubMed1989

Supraependymal cells and fibers in the third ventricle of the domestic chicken. A scanning electron microscopic study.

Supraependymal cells, fibers and what are presumed to be neuronal bulb-like projections were found in the third ventricle of the domestic chicken with a scanning electron microscope. At least two types of supraependymal cells were found: neuron-like cells and phagocyte-like cells. The former were predominantly seen in the area of the paraventricular organ and infundibular recess. The latter were abundant on the ventricular surface of the median eminence and subfornical organ. Bulb or club-like projections thought to be the dendritic terminals of CSF-contacting neurons were observed in the area of the paraventricular organ and infundibular recess. Similar structures were observed at the preoptic recess as well. The supraependymal neuronal components found in the domestic chicken differed from those of mammals in several respects: 1. the wall of the third ventricle was devoid of supraependymal fibrous plexus except for that of the paraventricular organ; 2. bulb-like projections were abundant in the area of the paraventricular organ; 3. supraependymal neuron-like cells were unipolar or bipolar in appearance. These data underline the dissimilarity of the CSF-contacting neuronal system of birds and mammals.

Animals↗
PubMed1988

Specialized ependyma in the posterior mesencephalon of the chicken: the fine structure of the subtrochlear organ.

A formation of specialized ependymal cells in the posterior mesencephalon of the domestic fowl, designated as the subtrochlear organ, was examined with light-, scanning- and transmission electron microscopy. This organ possessing the form of the letter "V" is located in the ventricular wall of the posterior mesencephalon. Its apex marks the median sulcus, while the arms of the V are directed rostrolaterally. Ependymal cells lining the subtrochlear organ usually project an extremely elongated process into the subependymal region and are classified into three types according to their surface features: (1) cells with a bulb-shaped protrusion that projects into the ventricle, (2) single cilium-bearing cells, and (3) cells with a tuft of cilia. The first type of cell is restricted to the median portion of the subtrochlear organ; its bulb-shaped protrusion contains numerous ribosomes. The second type of cell predominates in the arm (rostrolateral) area; in its apical cytoplasm such ciliary structures as basal body are rarely seen. The third type of cell is usually assembled into several small islands on the arm area; it has many basal bodies and other ciliary structures in the apical cytoplasm.

Animals↗