Search PubMed⌕ Search

Biomedical subjects

K Wake

Publications and source records attributed to K Wake.

At least 55 records · Page 3Linked to original sources

Perisinusoidal stellate cells of the liver: important roles in retinol metabolism and fibrosis.

In mammals, liver perisinusoidal stellate cells play an important role as a main store of body retinol (vitamin A). This fat-soluble vitamin is essential for vision, and regulates differentiation and growth of many cell types during embryonal development as well as in adult tissues. Thus, many cell types require a continuous supply of retinol. The storage of retinol (as retinyl esters) in stellate cells ascertains ample access of retinol to such cells also during periods with a low dietary intake. In lower vertebrates such as fish, vitamin A-storing stellate cells are found not only in the hepatic lobule, but also in the connective tissues of organs like intestine, kidney, ovaries, testes, and gills. Extrahepatic vitamin A-storing stellate cells are found in higher vertebrates when excessive doses of vitamin A are administered. It is not clear at present whether these cells also play a role in retinol metabolism under normal conditions. Stellate cells proliferate in a fibrotic liver, and they have been found to synthesize connective tissue compounds such as collagen. It was recently demonstrated that stellate cells are the principal cellular source of collagen and other extracellular substances in normal as well as fibrotic livers. Therefore, stellate cells, which seem to be a specialized type of pericyte, have a central role in the pathological changes observed during the development of liver fibrosis.

Animals↗

Liver units in three dimensions: I. Organization of argyrophilic connective tissue skeleton in porcine liver with particular reference to the "compound hepatic lobule".

Liver units were investigated in pig livers by means of histologic serial tracing, physical model building, and computer-aided three-dimensional imaging. Observations of the argyrophilic connective tissue skeleton were based mainly on the celloidin-embedded serial sections treated with silver impregnation. The parenchymal mass that clothed the initial segments of hepatic venous radicles was demarcated by fibrous septa which formed isolable units with two basic patterns: the simple hepatic lobule (SHL) and the compound hepatic lobule (CHL). Both lobule types presented regular limiting structures circumscribing each unit. Three-dimensional studies revealed that 25% of the lobules in a section belonged to the SHL type and 75% to the CHL type, the latter being predominant among the surface lobules. When considered in only two dimensions, however, the SHL-like lobules constituted the majority. Polygonal analysis disclosed that the pentagonal lobule was the most typical, instead of the "hexagonal" or "classic" lobule. The CHLs represented a multiaxial unit containing a system of venous tributaries in accordance with intralobular septation, whereas the SHLs were found with one axial vessel having a dendritic tendency at the incipient end; some SHLs were drained eccentrically by separate vessels into a sublobular vein. It was observed that, in dividing CHLs, whereas particular sinosoids were transformed into portal twigs, other sinusoids were changed into central venous tributaries. Fibrous deposition occurred along the septal-line sinusoids, bringing into view the septum-initiating plane. Fibroconnective tissue was supplied from the portal area and central (sublobular) adventitia, where portal triad structures and adventitial arterioles, respectively, were included. The findings of the present study facilitate the understanding of several characters of the lobules that have been reported previously, or occasionally postulated, such as the portal-central bridging tendency, the intralobular arterioles or ductules, the translobular artery or portal vein, the "portal-portal" or "portal-central" anastomoses, and the apposition of pericentral zone close to periportal zone. Based on differences in argyrophilia of sinusoidal reticulum, in proportion of lobule types, and in vasculature, the anatomic heterogeneity of liver unit was demonstrable in zonality, regionality, and locality.

Animals↗

Localization of hydroxyindole O-methyltransferase-synthesizing cells in bovine epithalamus: immunocytochemistry and in-situ hybridization.

Hydroxyindole O-methyltransferase (HIOMT)-immunoreactive cells and melatonin synthesis were demonstrated in bovine epithalamus (including the pineal gland) with monoclonal antibodies and cRNA probes to HIOMT. The HIOMT-immunoreactive product was present in the cytoplasm of pinealocytes. All identifiable pinealocytes were clearly labeled in the pineal gland. The expression of the HIOMT gene was first identified in pinealocyte cytoplasm by in-situ hybridization (ISH). The distribution of the hybridization-positive cells in the pineal gland was compatible with that revealed by immunocytochemistry using the monoclonal antibody to HIOMT. In addition, HIOMT transcripts were found in the medial habenular nucleus, and the habenular and posterior commissure; they may correspond to S-antigen-immunoreactive cells demonstrated in the same regions of the hamster and the mouse. In these regions, the hybridization-positive cells did not exhibit HIOMT-immunoreactivity; thus, cells devoid of immunoreactivity may synthesize but rapidly transport the newly synthesized proteins. These results indicate (1) that the conversion of N-acetylserotonin into melatonin takes place in the cytoplasm of pinealocytes, (2) that some epithalamic cells in the habenular area may synthesize melatonin, and (3) that melatonin may act as a chemical messenger in centrally directed processes, as shown by using S-antigen immunocytochemistry.

Acetylserotonin O-Methyltransferase↗

Localization and ultrastructure of the Kupffer cells in orthotopically transplanted liver grafts in the rat.

Kupffer cells play an important role in the acceptance or rejection of liver grafts. We examined the ultrastructure of the Kupffer cells in transplanted rat livers, from an early to a late stage where the graft is accepted, using a DA-to-PVG combination. Two days after surgery, endocytic activity of the Kupffer cells had increased, as evidenced by worm-like structures and many endocytic vacuoles. There was often close apposition to the monocytes or lymphocytes. By day 4, infiltration of mononuclear cells into the sinusoids was readily noticeable. By day 7, several Kupffer cells had migrated into the space of Disse through the openings in endothelial linings. The number of Kupffer cells reached a maximum at 14 days. They were located mostly outside the sinusoid, adhering to the hepatocytes. At this point in time, however, the Kupffer cells contained few endocytic vacuoles and phagolysosomes, in contrast to those at 2 days. The number and location of Kupffer cells became almost normal at 2 months. The present results indicate that Kupffer cells are highly activated before mononuclear cell infiltration becomes manifest in the sinusoid, and that when a rejecting reaction reaches a peak, they are usually located extrasinusoidally and show a morphologically immature profile.

Animals↗

Ultrastructural characteristics of lymphokine-activated killer cells of the rat in comparison with natural killer cells.

Lymphokine-activated killer (LAK) cells with a broad spectrum of tumor cell killing have been reported to be related to natural killer (NK) cells morphologically and phenotypically. We here examine the ultrastructural characteristics of LAK cells of the rat, comparing them to those of normal and OK-432-activated NK cells. Results show that, five days after the culturing of spleen lymphocytes with human recombinant interleukin-2, there were induced LAK cells, which were large granular lymphocytes and had a cytotoxic capacity against NK-resistant P-815 tumor cells. They were larger in size than NK cells and richer in cell organelles such as ribosomes, rough endoplasmic reticulum, a Golgi apparatus, granules and vesicles. The granules of LAK cells were shown to be related to multivesicular bodies as those of NK cells; they included multivesicular bodies, fully dense granules and intermediate forms between them. The average numbers and sizes of the granules and the proportion of multivesicular bodies and intermediate forms among the total granules were greater in LAK cells than in NK cells. The density of the small vesicles packed in multivesicular bodies and intermediate forms was much higher in LAK cells. At the contacting surface of the LAK cells bound to the target cells, exocytosis of multivesicular bodies was shown to occur. We recognized here two populations of LAK cells with different types of vesicles, one containing rod-cored vesicles and the other a new type of vesicles termed "demilune-cored vesicles". The latter vesicles were the same in size as the rod-cored ones and contained a dense core located eccentrically. Between these two populations of LAK cells, there was no difference concerning the profile of the dense granules. The present study indicates that, although LAK and NK cells share several ultrastructural features, the former show markedly enriched cell organelles, which indicate an accelerated metabolism of the cell for continuous proliferation.

Animals↗

The intracellular distribution of cell organelles in natural killer cells during the cytolysis of bound tumor cells, with special reference to the rod-cored vesicles.

The secretory function of the natural killer (NK) cells has been considered to be essential for cytolytic activity against their target cells. In this study, the distribution and spatial relationship of various cell organelles which participate in the secretory process, i.e., Golgi apparatus, vesicles, granules and microtubules, were examined ultrastructurally in non-treated and OK-432-activated rat NK cells bound to the tumor cells, with special reference to the rod-cored vesicles which are the most characteristic structure of the NK cells. Rod-cored vesicles and their closely related structures, "empty" vesicles, were derived from the end portion of the Golgi trans cisternae and became accumulated in the central area which was surrounded by the Golgi apparatus, nucleus and contact surface. Some of the vesicles appeared to be further transported to the contact area along the microtubules extending from the centrioles toward the bound targets. The access of the vesicles to the contact surface occurred at that portion where subplasmalemmal actin lattice was thin. The distribution of the dense granules and multivesicular bodies was similar to that of the vesicles, but the area of their occurrence was a little wider. At the outer aspect of the Golgi apparatus was situated the endoplasmic reticulum from which transitional vesicles came to the inner or cis cisternae of the apparatus. The present observations indicate that the cell organelles of the conjugated NK cells are purposefully arranged in the cytoplasm in such order that the generated rod-cored vesicles and "empty" ones are efficiently directed toward the bound tumor cells.

Animals↗

Stomatocytic or discoidal erythrocyte ghosts containing only spectrin.

We extracted Triton-treated erythrocyte ghosts with 2 M KCl (Triton/KCl/ghosts), and then with 1.2 M KBr at pH 5.5 (Triton/KCl/KBr ghosts). Triton/KCl/KBr ghosts were very similar in shape to untreated ghosts, Triton ghosts and Triton/KCl ghosts under a phase-contrast microscope at various pH vales and salt concentrations, despite having lost most of their phospholipids and proteins, except for spectrin. Negatively stained Triton ghosts, Triton/KCl ghosts and Triton/KCl/KBr ghosts appeared similar to each other, but the regularity of the spectrin network structure decreased somewhat in that order. Triton/KCl/KBr ghosts were stabilized by adding both actin and band 4.1, but not by adding either alone. These and previous findings strongly suggest that the spectrin network is visible and the simplest inframembrane structure.

Bromides↗

Differentiation of crypt epithelium in human palatine tonsils: the microenvironment of crypt epithelium as a lymphoepithelial organ.

The differentiation of the keratinocytes of the human palatine tonsils were studied by means of light and electron microscopy and immunohistochemistry using a polyclonal (K) and two monoclonal antikeratin antibodies (PKK1, PKK2). In the surface epithelium, the basal cells, cuboidal or columnar in shape, undergo progressive terminal differentiation and are transformed into the flattened cells of the upper layers. K reacts with both the basal and spinous layers, while PKK1 and PKK2 mark exclusively the basal layer. In the neck portion of the crypt, cavities containing one or aggregated lymphocytes with amorphous substances are observed in the spinous layer. The cavities are surrounded by elongated cytoplasmic processes of transformed epithelial cells bearing surface microvilli. These transformed epithelial cells display intense PKK1- and PKK2-positive reactions, whereas other conventional polygonal cells in the vicinity remain PKK1- and PKK2-negative as do those in the surface epithelium. In the deep portion of the crypt, where numerous lymphocytes invade the epithelium, the epithelial cells are transformed into star-shaped reticulum cells showing PKK1- and PKK2-positive reactions. The extended and branched cytoplasmic processes interconnect with one another constituting a complex network of reticulum cells, the well known reticulation of the crypt epithelium. Ten-nm filaments are usually oriented parallel to the longitudinal axis of transformed epithelial cells. Our observations suggest that the cell-shape transformation, i.e., from conventional polygonal epithelial cells into epithelial reticulum cells, occurs when the epithelial cells are in close contact with the infiltrating lymphocytes, and that this transformation is accompanied by a change in keratin phenotype.

Adolescent↗

24-hour rat liver preservation using UW solution and some simplified variants.

The results of a series of 32 rat liver transplants are described to analyze the efficacy of components of UW solution. Rat livers were stored at 4 degrees C in standard UW solution or one of three simplified variants for 24 hr prior to orthotopic liver transplantation. In standard UW solution (solution A) the one-week survival rate was 3 of 8. Using solution B, which differs from solution A in the omission of hydroxyethyl starch and adenosine, the one-week survival rate was 2 of 8. Solution C, a further-simplified version of solution B with omission of allopurinol, Bactrim, and insulin, gave a one-week survival rate of 3 of 8. Solution D is identical to solution B except that the sodium and potassium concentrations are reversed. Using this solution, 5 of 8 rats survived more than one week. We conclude that the effectiveness of UW solution is maintained in a substantially simplified form, and that solution D, with the Na/K ratio reversed to give a high Na variant, may improve survival.

Adenosine↗

Rhythm development in pineal and circulating serotonin, N-acetylserotonin, and melatonin in Syrian hamsters.

The ontogeny of diurnal rhythm patterns in the pineal and serum levels of melatonin, serotonin, and N-acetylserotonin was studied in Syrian hamsters (Mesocricetus auratus) from birth to adulthood. The pineal and blood specimens were collected at 1100 h and 0200 h, and the compounds were measured by radioimmunoassay (RIA) procedures. Pineal melatonin and serotonin did not show any circadian rhythm at day 5 of postnatal age. At this age N-acetylserotonin was undetectable in the light phase but became manifest at night. By 10 days of age pineal serotonin registered an established rhythm pattern, with a higher level during the day. The occurrence of circadian rhythm in pineal melatonin was delayed and manifested first at 25 days of age. At this age, the first detectable daytime level of N-acetylserotonin also occurred. Circadian rhythm in serum melatonin was also established at this age. The serum serotonin did not evince any rhythm pattern throughout the observation period, except at day 17 of postnatal age. The massive concentration of daytime serotonin in the pineal was not reflected in the circulatory system. For serum N-acetylserotonin there was no discernable day-night rhythm in all age groups, except at 25 days of age. The results show that the timing of the appearance of various compounds in the neonatal pineal is variable; the release of the substances does not always reflect their synthesis; the ontogenesis of circadian rhythm is a part of the maturational process; and 25 days of age is a rather critical time in development.

Animals↗

The neural organization of the pineal complex in the frog: stratification and regional differences.

Plexiform areas are functionally important units of the pineal complex. The silver impregnation technique by Matsuyama (1972) reveals in the frog that nerve fibres branch and terminate in these areas and that they are presynaptic to nerve cells which are part of the plexiform areas. Histochemically, the nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-diaphorase) method according to Scherer-Singler et al. (1983) indicates that pinealocytes and nerve cells are both stained, a strong stratification of different cell types can be visualized. Plexiform areas of different size exist at the dorsal and ventral surface of the organ; the biggest are located rostrodorsally. Approximately twenty large multipolar nerve cells are localized at the ventral surface of the pineal organ mainly at the rostral and lateral borders. They send long processes mainly in caudal direction. These processes are post-synaptic to the boutons of the endfeet belonging to photosensitive pinealocytes. Processes of several multipolar nerve cells converge to one plexiform area, branch and obviously terminate there. As in horizontal cells the processes of the multipolar cells seem to be capable of both receiving and transmitting signals. On the postsynaptic side of the processes of the multipolar cells are smaller nerve cells juxtaposed to the plexiform areas. They are known as pseudounipolar, unipolar or bipolar cells (Wake et al., 1974); the latter are free of boutons of the pinealocytes, their axons forming the pineal tract. Three different types of photosensitive pinealocytes can be distinguished: a slender, a spherical and a double-cone type. Also, the number of boutons varies in the endfeet: boutons occur alone, in groups of two or three, and in clusters.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Three-dimensional structure of endothelial cells in hepatic sinusoids of the rat as revealed by the Golgi method.

The three-dimensional structure of endothelial cells in the hepatic sinusoids of the rat was studied by application of light- and electron microscopy on Golgi-impregnated specimens. A number of endothelial cells could thus be individually delineated throughout the hepatic lobules. The cytoplasm, showing heavy silver deposits, consists of two distinct areas, a thick and thin portion. The thick portion, issuing from the region of the perikaryon, branches and tapers toward the cell periphery. The thin portion, occupying the remainder of the cytoplasm, consists largely of highly fenestrated sieve plates. Some intralobular variation can be noted; the thick portion of the endothelial cells is well developed in the periportal zone, while the cells in the centrilobular zone are relatively rich in thin portions. In addition, the area of distribution of an individual endothelial cell is larger in the centrilobular sinusoids than in the periportal zone. Some endothelial cells also possess unique cytoplasmic processes projecting into the intercellular space between hepatocytes and connecting the sinusoidal walls of neighboring sinusoids. These processes may anchor the endothelial cells to the hepatic plates.

Animals↗

Neuronal degeneration in the pineal ganglion during the post-hatching development of the domestic fowl.

The frequency of pineal ganglia associated with the pineal tract, and the numbers of acetylcholinesterase-positive neurons in these ganglia were studied in the domestic fowl during the post-hatching period by means of the acetylcholinesterase method. Furthermore, the degeneration of nerve cells in pineal ganglia of 40-day-old domestic fowl was investigated in detail at the electron-microscopic level. The rate of pineal organs containing one or more ganglia was 50% in 2- to 13-day-old, 38% in 40-day-old, and only 10% in 1-year-old domestic fowl. In parallel, the number of acetylcholinesterase-reactive nerve cells that constitute individual pineal ganglia decreased after hatching. Various degrees of neuronal degeneration were found in the pineal ganglia: swelling of the endoplasmic reticulum, electron-dense degeneration of the cytoplasm, and pyknosis of the nerve cell nucleus. Clusters of macrophages containing numerous lysosomes filled with debris-like material were scattered in the ganglion. In addition, plasma cells were observed in association with degenerating nerve cells. These results confirm the suggestion that the loss of acetylcholinesterase-positive nerve cells in the pineal ganglia of the domestic fowl is due to naturally occurring, programmed neuronal cell death. This process is discussed with reference to phenomena of cell death observed in other components of central nervous system.

Acetylcholinesterase↗

Changes in cell shapes and cytokeratins of epithelial cells during the infiltration of lymphocytes in the human palatine tonsils.

By the immunohistochemical method using anti-keratin antibodies (KL1-antigen, 56 KD keratin; PKK1-antigen, 40-52.5 KD keratin) and electron microscopy, the changes in cell shapes and the cytokeratin components of the epithelial cells during the infiltration of lymphocytes were studied. In the surface epithelium, PKK1 reacts with only the keratinocytes in the basal layer, while KL1 stains in the spinous layer. In the neck portion of the crypt, transformed keratinocytes scattered in the spinous layer react intensely with PKK1. These cells issue prolonged cytoplasmic processes which surround the cavities filled with infiltrating lymphocytes. In the deep portion of the crypt, PKK1-positive cells interconnect with one another constituting the network of the star-shaped reticular cells. The enlarged intercellular spaces of the reticular cells are filled with many lymphocytes. Our observations suggest that the infiltration of lymphocytes into the crypt epithelium induces the change in keratin expression of the epithelial cells in the human palatine tonsils, and the change of keratin molecules makes the stratified flattened epithelium to transform into the network of star-shaped reticular cells.

Child↗