Mineralocorticoid receptor blocker eplerenone improves endothelial function and inhibits Rho-associated kinase activity in patients with hypertension.

Hypertension is associated with endothelial dysfunction and activated Rho-associated kinases (ROCKs). The purpose of this study was to evaluate the effects of the selective mineralocorticoid receptor blocker, eplerenone, on endothelial function and ROCK activity in patients with hypertension. The study was carried out over 48 weeks in 60 untreated patients with hypertension who were randomly assigned to eplerenone, nifedipine, and losartan groups. We evaluated the effects of each treatment on flow-mediated vasodilation (FMD) and ROCK activity in peripheral leukocytes. Eplerenone increased FMD and decreased leukocyte ROCK activity. Nifedipine decreased ROCK activity but did not alter FMD. Losartan increased FMD but did not alter ROCK activity. Hypotensive effects were similar in the three groups, as was nitroglycerin-induced vasodilation during the follow-up period. There were no significant differences between the groups with respect to other parameters. The study results show that eplerenone improves endothelial function and inhibits ROCK activity in patients with essential hypertension.

The effect of proteasome inhibitor MG132 on experimental inflammatory bowel disease.

Immunoproteasome up-regulation enhances the processing of nuclear factor-kappaB (NF-kappaB) and degradation of IkappaBalpha, which correlates with increased amounts of NF-kappaB in the various cells. Aberrant activation of NF-kappaB is involved in the pathogenesis of inflammatory bowel disease (IBD). The aim of this study was to elucidate the effect of proteasome inhibitor MG132 on experimental IBD. We investigated the effects of MG132 on intestinal inflammation and epithelial regeneration in both interleukin-10-deficient (IL-10(-/-)) mice and mice with dextran sulphate sodium (DSS)-induced colitis. Body weight, histological findings and tumour necrosis factor (TNF)-alpha mRNA expression, epithelial cell proliferation and NF-kappaB p65 activity in colonic tissues were examined. The effects of MG132 on cell proliferation, migration and multiple drug resistance 1 (MDR1) gene expression were determined in vitro. MG132 ameliorated intestinal inflammation of IL-10(-/-) mice by decreasing TNF-alpha mRNA expression in the colonic tissues, which was associated with suppression of NF-kappaB activation, and reduced significantly the number of Ki-67-positive intestinal epithelial cells. On the other hand, MG132 did not reduce intestinal inflammation in mice with DSS-induced colitis, and delayed significantly the recovery of body weight and epithelial regeneration. MG132 also suppressed significantly epithelial cell proliferation, cell migration and MDR1 gene expression in vitro. Proteasome inhibition reduces T cell-mediated intestinal inflammation, but may interrupt both epithelial regeneration and barrier function of colonic mucosa. Optimal use of proteasome inhibitor should be kept in mind when we consider its clinical application for patients with IBD.

Effects of vitamin A deficiency on the function of pituitary-gonadal system in male rats.

The effects of vitamin A deficiency on the pituitary-gonadal function were examined by measurements of serum and pituitary level of pituitary hormones and serum testosterone concentration, and by investigations of histological changes in the testis and the pituitary gland in vitamin A-deficient (VAD) and supplemented (VAS) rats. The growth of VAD rats was retarded and their body weights were decreased after 9 weeks of experiments and attained about one half of the weight of control animals at 12 weeks. In the VAD rats, serum testosterone concentrations were decreased significantly compared with those in the VAS controls. Serum and pituitary concentrations of GH were significantly lower but those of LH were slightly lower in the VAD rats than those in the controls, while the serum FSH concentration was significantly higher than that in the control rats. The seminiferous tubules in the testes of VAD rats were comprised largely of Sertoli cells and a reduced number of spermatogonia and contained fibrous formation in their lumen. In the pituitary gland, GH cells were significantly reduced in number in the VAD rats, but gonadotropic (GTH) cells were increased remarkably in size and number, showing hypertrophy and vacuolation similar to those in castration cells. The cytological changes in the pituitary gland and the increased discharge of FSH represent a secondary and compensatory change similar to that seen following castration and vitamin E deficiency.

Localization of avian LHRH-immunoreactive neurons in the hypothalamus of the domestic fowl, Gallus domesticus, and the Japanese quail, Coturnix coturnix.

The localization of LHRH-containing perikarya and nerve fibers in the hypothalami of the domestic fowl and Japanese quail was investigated by means of the specific immunoperoxidase ABC method, using antisera against chicken LHRH-I ([Gln8]-LHRH), chicken GnRH-II ([His5-Trp7-Tyr8]-LHRH[2-10]) and mammalian LHRH ([Arg8]-LHRH). Chicken LHRH-I-immunoreactive perikarya were sparsely scattered in the nucleus preopticus periventricularis (POP), nucleus filiformis (FIL) and nucleus septalis medialis (SM), and in bilateral bands extending from these nuclei into the septal area in both species. A few reactive perikarya were also observed in the nucleus accumbens (Ac) and lobus parolfactorius (LPO). Numerous cLHRH-I-immunoreactive fibers were widely scattered in the preoptic, septal and tuberal areas, and were densely concentrated in the external layer of the median eminence and in organum vasculosum of the lamina terminalis (OVLT) in both species. Anti-mammalian LHRH serum cross-reacted weakly with perikarya and fibers immunoreactive to anti-cLHRH-I serum in normal chicken and quail. Anti-cGnRH-II[2-10] serum immunoreacted with magnocellular neurons distributed in the rostral end of the mesencephalon along the midline close to the nervus oculomotorius (N III). These perikarya were apparently different from cLHRH-I immunoreactive neurons. No immunoreactive cells and fibers against anti-cGnRH-II[2-10] were observed in the hypothalamus and median eminence of the chicken or quail. Anti-cGnRH-II[2-10] bound specifically with cGnRH-II.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of vitamin E deficiency on the hormone secretion of the pituitary-gonadal axis of the rat.

Chronological changes of gonadotropin (FSH and LH) and testosterone concentrations in the serum were measured in vitamin E deficient rats to investigate the effects of vitamin E deficiency on the pituitary-gonadal function in rats. The receptor sites and association constant (Ka) for LH and the formation of cyclic AMP in the Leydig cells were also investigated. The results obtained in the present study are as follows: 1) The vitamin E deficient rats showed almost complete hemolysis and extremely increased TBA reacting substances (TBARS) in the serum and liver. 2) The serum LH concentration in the vitamin E deficient group was slightly higher than in the vitamin E supplemented group during the later periods of experiment. 3) The serum FSH concentration in the vitamin E deficient group did not differ significantly from that in vitamin E supplemented group, but became significantly higher than that in the latter at 186 days of experiment. 4) The serum testosterone concentration was always lower in the vitamin E deficient group than in the control. 5) The vitamin E deficient group showed slightly large number of LH/hCG receptor and significantly small Ka (low affinity), as compared with vitamin E supplemented group. The formation of cyclic AMP by Leydig cells decreased significantly in vitamin E deficient group. These results suggest that the vitamin E deficiency exerted a suppressive effect directly on the gonadal function to decrease the hormone synthesis in the Leydig cells and caused the increased secretion of pituitary LH owing to the feedback mechanism.

Immunohistochemical localization of corticotropin-releasing factor (CRF)-containing neurons in the hypothalamus of the Japanese quail, Coturnix coturnix.

Immunohistochemical localization of corticotropin-releasing factor (CRF)-like immunoreactivity in the brain of the Japanese quail was studied by means of the peroxidase anti-peroxidase (PAP) method. CRF-immunopositive perikarya of parvocellular neurons were observed mainly in the nucleus praeopticus medialis and nucleus paraventricularis. Additional perikarya were also detected in the nucleus hypothalamicus posterior medialis in the hypothalamus and in the non-hypothalamic nucleus accumbens, nucleus septalis lateralis and nucleus dorsomedialis and dorsolateralis thalami. No CRF immunoreaction was found to coexist with the vasotocin (Vt)-containing system in comparative examination of consecutive sections treated with anti-vasopressin (Vp) serum. The CRF-immunoreactive fibers were detected mainly in the external layer of the anterior median eminence but not in its posterior division. Unilateral adrenalectomy induced the marked reduction in number of the CRF immunopositive fibers in the anterior median eminence.

Immunohistochemistry of the hypothalamic neuropeptides and anterior pituitary cells in the Japanese quail.

The pars distalis of the avian adenohypophysis consists of well-defined cephalic and caudal lobes which are distinct in their cellular constituents. Immunocytochemical investigations on the pituitary hormones of the pars distalis of the Japanese quail reveal five types of secretory cells, adenocorticotropin (ACTH) cells, prolactin (PRL) cells, thyroid-stimulating hormone (TSH) cells, growth hormone GH (STH) cells, and FSH/LH (gonadotropic) cells. The ACTH cells, TSH cells, and PRL cells are restricted to the cephalic lobe, and GH (STH) cells are confined to the caudal lobe, while FSH/LH cells are distributed throughout the cephalic and caudal lobes. The median eminence of birds has distinct anterior and posterior divisions, each with different neuronal components. The avian hypophysial portal vessels also consists of two groups, anterior and posterior. The peculiar arrangement and distribution of the avian hypophysial portal vessels are possibly related to the distribution of neuropeptides in the two divisions of the median eminence and to the cytological and functional differentiation of two lobes of the pars distalis. The localization of perikarya and fibers containing luteinizing hormone releasing hormone (LHRH), somatostatin, vasotocin, mesotocin, corticotropin-releasing factor (CRF), vasoactive intestinal polypeptide (VIP), glucagon, metenkephalin, and substance P in the hypothalamus and median eminence of the Japanese quail has been investigated by means of immunohistochemistry using antisera against the respective neuropeptides. LHRH-, somatostatin-, VIP-, met-enkephalin-, and substance P-immunoreactive fibers are localized in the external layer of the anterior and posterior divisions of the median eminence, while CRF- and vasotocin-reactive fibers are demonstrated only in the external layer of the anterior division of the median eminence. The metenkephalin fibers are thicker in the anterior median eminence but the substance P fibers are more abundant in the posterior division. Mesotocin fibers occur only in the internal layer of the median eminence and neural lobe.

Immunohistochemical localization of vasoactive intestinal polypeptide (VIP)-containing neurons in the hypothalamus of the Japanese quail, Coturnix coturnix.

The localization of vasoactive intestinal polypeptide (VIP) in the hypothalamus of the quail has been studied by means of light- and electron-microscopic immunohistochemistry. Numerous VIP-immunoreactive perikarya are distributed in the caudal portion of the nucleus infundibularis (n. tuberis) and nucleus mamillaris lateralis, and sparse in the preoptic area, nucleus supraopticus and nucleus paraventricularis. Dense localization of immunoreactive-VIP fibers is observed in the external layer of the median eminence, in close contact with the primary portal capillaries. The main origins of these fiber terminals are VIP-immunoreactive perikarya of the nucleus infundibularis. These neurons are spindle or bipolar and extend one process to the ventricular surface and another to the external layer of median eminence. They are CSF-contacting neurons and apparently constitute the tuberohypophysial tract that links the third ventricle and the hypophysial portal circulation. VIP-reactive neurons in the nucleus mamillaris lateralis also project axons to the external layer of the median eminence, constituting the posterior bundle of the tuberohypophysial tract. Numerous VIP-immunoreactive perikarya occur also in the nucleus accumbens/pars posterior close to the lateral ventricle. They are also CSF-contacting neurons extending a process to the lateral ventricle. There are moderate distributions of VIP-reactive fibers in the area ventralis and in the area septalis. Ultrastructurally, the immunoreactive products against VIP are found in the elementary granules, 75-115 nm in diameter, within the nerve fibers in the median eminence.

Immunocytochemical localization of neurotensin-containing neurons in the hypothalamus of the Japanese quail, Coturnix coturnix japonica.

The hypothalamus of Japanese quail, Coturnix coturnix japonica, has been studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibodies to synthetic neurotensin (NT). A number of immunoreactive neuronal perikarya occur in the medial preoptic nucleus of the rostral hypothalamus and a few in the accessory part of paraventricular nucleus and dorsal portion of the infundibular nucleus. Some of them correspond to the parvocellular neurons previously identified tentatively as neurosecretory (Mikami et al. 1975, 1976). Large numbers of immunoreactive neuronal fibers are found in the preoptic area, which extend as a remarkable fiber tract from this area to the ventral septal area and to the subfornical organ. A few immunoreactive fibers also extend ventrocaudally to the infundibular nucleus and to the neural lobe.

The hypothalamic neurosecretory systems of the Japanese quail as revealed by retrograde transport of horseradish peroxidase.

Within 24-48 h after injection of horseradish peroxidase (HRP) into the neural lobe or into the median eminence of adult Japanese quail dense accumulations of its reaction product (HRP-RP) can be demonstrated in axons of the hypothalamo-hypophysial tract and in the magnocellular neurosecretory perikarya of the supraoptic and paraventricular nuclei as well as in scattered neurons of the accessory hypothalamic neurosecretory nuclei. The HRP-RP-containing nerve fibers, which are beaded in appearance, occur prominently in the internal zone of the median eminence. They turn dorsally at its anterior border to become widely distributed in the retrochiasmatic region and extended to the paraventricular, supraoptic areas. These observations confirm more directly conclusions drawn earlier from Gomori-type preparations and from immunologic demonstration of arginine vasotocin, mesotocin and neurophysin. HRP-RP was also found in perikarya of parvocellular secretory neurons in the infundibular nucleus 48 h after injection of HRP into the median eminence but not after injection into the pars nervosa. This provides direct evidence that a conspicuous component of the tubero-infundibular tract is formed by axons of tuberal neurons that originate from the infundibular nucleus and pass directly into the median eminence.

Ultrastructure of the organum vasculosum of the lamina terminalis of the Japanese quail, Coturnix coturnix japonica.

The structure of the organum vasculosum of the lamina terminalis (OVLT) of the Japanese quail has been studied by light and electron microscopy. The OVLT has a palisade arrangement. It forms a part of the terminal plate extending from the optic chiasma to the anterior commissure and is characterized by a special vascular arrangement. The organ consists of ependymal, internal and external zones. The ventricular surface of the organ is covered by non-ciliated ependymal cells characterized by the presence of raspberry-like ventricular protrusions. The ependymal zone is composed of two types of ependymal cells, one being a large, cuboidal cell with columinous cytoplasmic protrusions suggesting secretion into ventricle, and the other a slender tanycyte with long balar plexus of the external surface. The internal zone contains two types of secretory neurons, parvocellular neurons containing a few dense-cored granules 1,000 A in diameter, and mediocellular neurons containing in their perikarya many larger granules 1,300-1,500 A in diameter and well developed granular endoplasmic reticulum and Golgi apparatus. The granular formations are usually observed in the Golgi area of both types of cells, thus indicating of their secretory activities. There are three types of nerve terminals in the neuropil: (1) nerve endings containing spherical, dense-cored granules 800 A in diameter, which display axosomatic synapses with perikarya of the neurons, (2) nerve endings containing dense-cored granules 1,000 A in diameter and clear vesicles 400 A in diameter, and (3) nerve endings containing dense granules 1,300 A in diameter and clear vesicles. Types 2 and 3 do not display axo-xomatic synapses but often show axo-dendritic synaptic contacts with dendrites in the neuropil; thus they are probably axons originating from the parvocellular and mediocellular neurons of this organ. The functional significance of the neurons and axons of the OVLT is discussed and it is suggested that two kinds of neurohormones may be secreted from the OVLT of the Japanese quail.

The fine structure of the hypothalamic secretory neurons of the white-crowned sparrow, Zonotrichia leucophrys gambelii (Passeriformes: Fringillidae). II. Magnocellular and parvocellular nuclei of the rostral hypothalamus.

The fine structures of the neurons and neuropils of the magnocellular supraoptic nucleus and the parvocellular nuclei of the rostral hypothalamus, including the suprachiasmatic and medial, lateral and periventricular preoptic nuclei, and the neuronal apparatus of the organum vasculosum laminae terminalis, have been examined in the male White-crowned Sparrow, Zonotrichia leucophrys gambelii, by correlated light and electron microscopy. The magnocellular supraoptic nucleus is characterized by large neurosecretory perikarya which contain a well developed Golgi complex and dense-cored granules 1,500-2,200 A in diameter. The neuropil displays axons, dendrites and glial fibers. Some axonal profiles contain dense-cored vesicles 800-1,000 A in diameter and clear vesicles 500 A in diameter. Axo-somatic and axo-dendritic synapses are conspicuous in this nuclear region. The suprachiasmatic nucleus is characterized by an accumulation of small neurons with moderately developed cellular organelles and some dense-cored granules, approximately 1,000 A in diameter. The profiles of axons within the neuropil contain dense-cored granules 800-1,000 A in diameter and clear vesicles 500 A in diameter. The neurons of the medial preoptic nucleus are relatively large and exhibit well developed cellular organelles and dense-cored granules 1,300 to 1,500 A in diameter. Granular materials are formed within the Golgi complex. The medial preoptic nucleus is rich in secretory perikarya. Occasionally, neurons with granules 1,500-2,200 A in diameter are encountered in the lateral preoptic and periventricular preoptic nuclei. They may be considered as scattered elements of the magnocellular (supraoptic and paraventricular) system. The organum vasculosum laminae terminalis consists of three layers, i.e., ependymal, internal and external zones, and exhibits a vascular arrangement similar to that of the median eminence. The perikarya of the parvocellular neurons and their axons in the internal zone contain numerous secretory granules ranging from 1,300 to 1,500 A in diameter.

The fine structure of the hypothalamic secretory neurons of the white-crowned sparrow, Zonotrichia leucophrys gambelii (Passeriformes: Fringillidae). I. Parvocellular tuberal nuclei.

The fine structure of the parvocellular tuberal nuclei and that of the ependyma bordering the third ventricle in the basal hypothalamus of the White-crowned Sparrow, Zonotrichia leucophrys gambelii, have been investigated. Photoperiodically stimulated birds have been compared with birds held on short days. The perikarya of the neurons of the basal infundibular (tuberal) nucleus, and in part, of the more dorsal layers, contain dense-cored granules (1000-1500 A). The granules in the anterior part of the nucleus are somewhat larger than those of the posterior part. The synapses and the synaptic relationships of these cells are described. The single-layered ependyma of the third ventricle in the basal hypothalamus may be divided into the dorsal typical ependyma, the ventrolateral "glandular" ependyma, and the ventral "glandular" ependyma. Cells of the ventral ependyma lack apical cilia but bear a few microvillous processes. They have well-developed Golgi apparatus, conspicuous polysomes, and frequently dense, irregularly-shaped granules. Basal cytoplasmic processes extend ventrally to the outer surface of the median eminence. Photoperiodic stimulation appears to increase the numbers of apical protrusions of the cells in the ventral glandular ependyma and to cause an increase in size of the nerve cells of the basal infundibular nucleus.