Ultrastructural demonstration of exocytosis in the intact rat adrenal medulla

Evidence is presented for morphological proof of exocytosis in the rat adrenal medulla in situ. Techniques were modified to allow perfusion of the intact adrenal gland with secretagogues (or electrical stimulation) followed by tannic acid. Unstimulated specimens demonstrated exocytotic (omega-shaped) profiles filled with flocculent material. This flocculation was also seen in the intercellular space. Stimulation of the adrenal medulla also resulted in the appearance of exocytotic profiles and an accumulation of the flocculent mass. This was often most evident in the subendothelial space. This is the first demonstration of exocytosis in the rat adrenal medulla by electron microscopy. The techniques used in this study will be useful for studying the pathway of secretory products of the adrenal chromaffin cell before they enter the vascular system.     

Immunocytochemical localization of neuropeptides in the adrenal medulla

The distribution of neuropeptides exhibits pronounced interspecies heterogeneity. Neuropeptides may function as hormones secreted from chromaffin cells or as neurotransmitters/neuromodulators released from nerve terminals. However, other possible functions such as trophic or intracellular effects should also be considered. Thus, to understand the role of neuropeptides, it is important to explore their localization in different species. The distribution of enkephalins, neurotensin, neuropeptide Y, calcitonin gene-related peptide, and galanin in the adrenal medulla of rat, cat, hamster, and mouse is presented in detail.     

Electron microscopic localization of enkephalin-like immunoreactivity in the human adrenal medulla

Enkephalin-like immunoreactivity in human adrenomedullary cells was studied at the light and electron microscopic levels. Enkephalin immunostaining was associated with chromaffin granules and, in a few cells, with the rough endoplasmic reticulum as well. The relative number of stained granules varied from cell to cell, and a correlation with a particular granular population was not noted. Both large and small granules were labelled. It is concluded that in the human the ability to store enkephalin immunoreactive peptides is a general property of chromaffin granules and, furthermore, is not correlated with specific granular subpopulations or the particular type of catecholamine stored within the cell.     

A fluorescence histochemical study of ketosteroids in gonadal and adrenal tissue of the rat

Sections of formalin-fixed gonads from normal rats, and adrenals and ovarian transplants from normal and castrated rats, were treated with salicylhydrazide (after pretreatment with methylhydrazine and sulphobenzaldehyde) and examined for fluorescence before and after mounting in alkali. Sites containing ketosteroids demonstrable in this way are described. Evidence is presented which suggests that luteinized thecal cells of non-cyclic ovarian transplants may secrete androgens. Ovarian tissue which has been transplanted to male rats castrated at birth behaves as in the normal female (Pfeiffer, 1936; Yazaki, 1960; Harris, 1963, 1964). Its activity does not appear to depend on the adrenal, and castration of the newborn male rat does not lead to any obvious histological change in the adrenal (Adams Smith, 1966). However, production of androgen has been observed, using biological criteria, in ovaries grafted into the ears of castrated mice (Hill, 1937a, b) and rats (Deanesly, 1938). It therefore seemed worthwhile investigating the ketosteroid content of such tissues histochemically. In this paper the histological distribution of two different types of ketosteroid in gonads of normal rats, in adrenals of normal and castrated male rats and in ovarian transplants in the eyes of male rats (either castrated at birth or at two months of age, or with intact testes) will be described. The ketosteroids were detected histochemically by condensing them with salicylhydrazide (after pretreatment with methylhydrazine and sulphobenzaldehyde) and then observing the fluoresence they emit before and after treatment with alkali (Stoward & Adams Smith, 1964).     

Cytochrome b558 (p22phox) in the guinea-pig adrenal medulla.

Paraganglionic cells are sensitive to hypoxia, and the involvement of a plasmalemmal cytochrome b558-like protein in oxygen sensing by these cells has been suggested, but neither the identity of the immunoreactive protein detected by immunohistochemistry nor its anticipated subcellular (i.e., plasmalemmal) localization were directly proven. Thus, we extended these studies to the largest paraganglion, i.e., the adrenal medulla, in the guinea-pig, which, due to its size and accessibility, allowed us to address both of these issues utilizing antisera raised against synthetic peptides of the small (22 kD) subunit of cytochrome b558, p22phox. Cytochrome b558 was originally identified in granulocytes and macrophages, and antisera against this phagocyte p22phox were utilized. Immunoreactivity to p22phox was observed in all adrenal medullary endocrine cells, and the identity of the immunoreactive protein to the small cytochrome b558-subunit was confirmed by Western blotting. Immuno-electron microscopy of ultrathin cryosections and of resin-embedded tissue demonstrated its subcellular localization in the dense core vesicles of endocrine A-cells but not in the plasma membrane. In conclusion, the present study documents the presence of the small subunit of cytochrome b558 in guinea-pig adrenal medullary cells, but its subcellular vesicular localization does not support the initial interpretation of cytochrome b558 serving as a plasmalemmal oxygen sensor.     

An experimental study on the temperature distribution in IRWST

The In-Containment Refueling Water Storage Tank (IRWST), one of the design improve-ments applied to the APR-1400, has a function to condense the high enthalpy fluid discharged from the Reactor Coolant System (RCS). The condensation of discharged fluid by the tank water drives the tank temperature high and causes oscillatory condensation. Also if the tank cooling water temperature approaches the saturated state, the steam bubble may escape from the water uncondensed. These oscillatory condensation and bubble escape would burden the undue load to the tank structure, pressurize the tank, and degrade its intended function. For these reasons simple analytical modeling and experimental works were performed in order to predict exact tank temperature distribution and to find the effective cooling method to keep the tank temperature below the bubble escape limit (93.3°C), which was experimentally proven by other researchers. Both the analytical model and experimental results show that the temperature distributions are horizontally stratified. Particularly, the hot liquid produced by the condensation around the sparger holes goes up straight like a thermal plume. Also, the momentum of the discharged fluid is not so strong to interrupt this horizontal thermal stratification significantly. Therefore the layout and shape of sparger is not so important as long as the location of the sparger hole is sufficiently close to the bottom of the tank. Finally, for the effective tank cooling it is recommended that the locations of the discharge and intake lines of the cooling system be cautiously selected considering the temperature distribution, the water level change, and the cooling effectiveness.     

An experimental study of contact pressure distribution in panel stamping operations

This paper presents an analysis of the dynamic contact pressure distribution at the sheet metal-tooling interface in normal and defective panel stamping operations. Specifically, the stamping of a 1.27?×?20?×?15?cm rectangular aluminum alloy panel has been simulated using finite-element modeling methods. The modeling results are leveraged to determine the forces acting on sensors embedded in the die structure. Based on these results, a customized stamping test bed has been designed, which incorporates an array of force sensors embedded into the stamping die to enable online measurement of the forces in the die. The force measurements are processed by the thin plate spline numeric surface interpolation method to evaluate a continuous contact pressure distribution map of the die cavity. The pressure distribution in the die cavity in normal and defective scenarios has been presented along with a suggested volume thresholding method for automated defect detection. By leveraging the numeric surface generation method to analyze experimental data, the present study establishes the basis for intelligent tooling and in-process monitoring for improved observability in sheet metal stamping.     

Immunohistochemical study on the distribution of sodium-dependent vitamin C transporters in the respiratory system of adult rat

As vitamin C (L-ascorbic acid, VC) is known to be essential for many enzymatic reactions, the study on the transport mechanism of VC through cytoplasmic membrane is crucial to understanding physiological role of VC in cells and the respiratory system. In this regard, the study on the newly identified sodium-dependent VC transporters (SVCTs), SVCT1 and SVCT2, is required in organs that contain high concentration of VC. We have shown the distribution of SVCT proteins in the respiratory system, which has been reported to be one of the organs with a high concentration of VC, using immunohistochemical techniques. In the present study, intense SVCT immunoreactivities (IRs) were mainly localized in the respiratory system epithelial cells. In the trachea, both SVCT1 and 2 were localized in the psuedostratified ciliated columnar epithelium. In the terminal bronchiole, SVCT1 and 2 IRs were mainly observed in the apical portion of the simple columnar epithelium. In addition, SVCT IRs was localized within the cell membrane of some alveolar cells, even though we could not identify the exact cell types. These results provide the first evidence that intense SVCT1 and 2 IRs were found in the apical portion of the respiratory epithelial cells, suggesting that SVCT proteins in the apical portion could transport the reduced form of VC included in the airway surface liquid into the respiratory epithelial cells.     

An experimental study on the distribution of the drop size and velocity in asymmetric impinging jet sprays

Distributions of the drop size and velocity in an asymmetric impinging jet are investigated by injecting water and a sodium carbonate (Na₂CO₃) solution, which simulates the mixing process in impinging jet sprays of liquid oxidizer and liquid fuel for liquid propellants. The liquid sheet formed from the impinging jet is visualized and the drop size distributions are obtained by using image processing for the visualized images. The drop size distribution of the asymmetric impinging jets is fitted to the Rosin-Rammler distribution function. The obtained drop size distributions according to the azimuth angle in the impinging jet are compared with the theoretical predictions of previous research. The experimental results of the drop size distributions are located between the two curves obtained from the theoretical predictions by treating each jet in the asymmetric impinging jets as an independent wall-impinging jet. PIV images using a double-exposure method were processed to obtain the drop velocity vector in the impinging jets. Whether the drops shedding from the edge of the asymmetric impinging jets occurs radially or tangentially is also investigated from the PIV results.     

An experimental study on the improvement of mixture distribution for the automotive S.I. Engines

In a fuel supply system of gasoline engines, a part of the fuel ejected from the carburetor floats in flow with high speed as dro plets in an intake manifold and the rest of it flows as the liquid film along the wall of the intake manifold and is gradually vaporized in the flow. Finally, most part of the fuel terminates the vaporization after introducing mixture into cylinders. Accordingly, it is desirable to maintain the optimum mixture ratio in conventional carburetor systems for all operating conditions of engines. The intake system including the carburetor of automotive gasoline engines which are mostly operating in partload conditions have many unsolved problms, such as the increase of specific fuel consumption and reduction of exhaust emissions due to the rich mixture in starting, acceleration and over load operations. Therefore the main purpose of the study is to develop a special device (so called Eco-Ring) for the enhancement of atomization that has ability to separate the liquid film spray flowing along the pipe wall of the intake manifold and to atomize it to fine droplets, then the device can overcome some defects which arise in the conventional carburetor systems as mentioned above.     

Immunohistochemical study of heat shock proteins 27, 60 and 70 in the normal human adrenal and in adrenal tumors with suppressed ACTH production

Heat shock proteins (HSPs) are known to protect cells against various aggressions and to assist in the correct folding of nascent proteins as well as in the recovery of denatured ones. HSP70 increases its levels in the cell in response to any stress and is induced by ACTH in the adrenal gland. HSP60 is located in the mitochondria and assists in the folding of mitochondrial peptides. HSP27 is the only small HSP that is stress-induced. HSP27 and HSP70 are known to protect cells against apoptosis while, on the contrary, HSP60 is proapoptotic, increasing caspases maturation. We studied the expression of these HSPs in human adrenal tissue both in the normal glands (12 cases) and in tumoral tissue from cortisol producing adrenal adenomas (6 cases). Besides being neoplastic, these cells live in a particular ambience of lack of ACTH due to the suppression of the hypothalamic-pituitary ACTH secretion induced by the elevated levels of cortisol. HSP27 is highly expressed in the normal adrenal and shows a marked reduction of expression in Cushing's adrenal tissue. Although with overall lower levels of expression in the normal adrenal, HSP70 exhibited a similar pattern of reduction in tumoral tissue. HSP60, on the other hand, increased significantly and consistently in adrenal Cushing tumors. Besides the possible consequences of incorrect folding of nascent peptides, the alterations observed in tumoral tissue seem to act in an apoptotic direction. The only factor that we observed that could be contributing to these changes was the lack of plasma ACTH.     

Spatial distribution of Reissner's fiber glycoproteins in the filum terminale of the rat and rabbit

The subcommissural organ secretes into the third ventricle glycoproteins that condense to form the Reissner's fiber (RF). At the distal end of the central canal of the spinal cord, the RF-glycoproteins accumulate in the form of an irregular mass known as massa caudalis. Antibodies against RF-glycoproteins and a set of lectins were used at the light and electron microscopic level to investigate the spatial distribution of the massa caudalis material in the rat and rabbit filum terminale. In the sacral region of the rat, the central canal presents gaps between the ependymal cells through which RF-glycoproteins spread out. The bulk of massa caudalis material, however, escapes through openings in the dorsal wall of the terminal ventricle. In the rabbit, the massa caudalis is formed within the ependymal canal, at the level of the second coccygeal vertebra, it accumulates within preterminal and terminal dilatations of the central canal, and it escapes out through gaps in the dorsal ependymal wall of the terminal ventricle. The existence of wide intercellular spaces and a large orifice (neuroporous) in the dorsal ependymal wall of the terminal ventricle, and the passage of RF-material through them, appear to be conserved evolutionary features. After leaving the terminal ventricle of the rat and rabbit, RF-glycoproteins establish a close spatial association with the numerous blood vessels irrigating the filum terminale, suggesting that in these species the blood vessels are the site of destination of the RF-glycoproteins escaping from the central canal, thus resembling the situation found in lower vertebrates. When passing from the RF stage to the massa caudalis stage, the rabbit RF-glycoproteins lose their sialic acid residues, exposing galactose as the terminal residue. Since this sialic acid-galactose modification of RF-glycoproteins had also been described in lamprey larvae, it may be regarded as a conserved evolutionary feature associated with the formation of the massa caudalis.     

Involvement of the adrenal glands and testis in gap junction formation via testosterone within the male rat anterior pituitary gland

We investigated the influence of testicular and adrenal androgens on the presence of gap junctions between folliculo‐stellate cells in the anterior pituitary glands of 60‐day‐old Wistar‐Imamichi strain male rats. The animals were separated into six groups: Group A served as the controls and had free access to a normal diet and water, Group B was given a normal diet and 0.9% NaCl for their drinking water as the controls of adrenalectomized groups, Group C was castrated, Group D was adrenalectomized, Group E was both castrated and adrenalectomized, and Group F was also both castrated and adrenalectomized. In addition, the animals of Group F were administered a dose of testosterone that is known to produce high physiological levels of the hormones in plasma. Five rats from each group were sacrificed 1, 2, 3, 4, 5, 6, and 7 days after their respective operation, and the anterior pituitary glands were removed and prepared for observation by transmission electron microscopy. We quantified the number of follicles and gap junctions and calculated the rate of occurrence as the ratio of the number of gap junctions existing between folliculo‐stellate cells per intersected follicle profile. Simultaneous removal of adrenal glands with castration resulted in a significantly decrease in the number of gap junctions, whereas the administration of testosterone to these rats compensated for this change. These observations indicate that the preservation of gap junctions between folliculo‐stellate cells is mainly dependent on androgens from both the testes and adrenal glands in adult male rats. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

An investigation of velocity distribution in rotating magnetohydrodynamic bearings

An analysis based on the Navier-Stokes equations is made for the flow of a liquid in magnetohydrodynamic circular bearings under rotating conditions. The analysis presents expressions for both radial and tangential velocities with and without inertia effects. The results show that the application of a magnetic field reduces both radial and tangential velocities. The rotational inertia effect has a greater effect on the radial velocity at small film thicknesses.     

An autonomous digital complex for measuring the temperature distribution

An autonomous measuring complex has been designed for monitoring the distributed temperature field in the sea. The system includes a control and data storing module, a power supply, a timing system, and peripheral devices (DS-18B20 digital temperature sensors). The complex is based on the modular structure, which allows various configurations of sensors to be produced for each particular task with minimum expenses.