Capillary Lumina Research Articles

Characterization of the Avian Polyomavirus-Associated Glomerulopathy of Nestling Parrots

The glomerulopathy occurring in nestling nonbudgerigar parrots with avian polyomavirus (APV) disease was examined in 10 parrots. Glomerular lesions were characterized by the presence of dense, periodic acid-Schiff (PAS)-positive material that expanded the mesangium and that narrowed and at times occluded capillary lumina. PAS-staining was found to be more sensitive than hematoxylin and eosin in the detection of the lesions. Ultrastructurally, finely granular electron-dense material was found in massive intracapillary and mesangial condensates. Capillary endothelial cells exhibited changes consistent with cellular swelling. No evidence of chronic glomerular changes was observed. Immunofluorescent staining demonstrated that the PAS-positive, electron-dense condensates were complexes of IgG, avian polyomavirus antigen, and, in one bird, IgM. Viral DNA was detected in the serum of all six birds examined. Anti-APV antibodies were also present in all five serum samples examined. These findings suggested that the pathogenesis of this acute immune complex glomerulopathy and other APV-associated lesions depends on the presence of an appropriate ratio of circulating virus and anti-APV antibody.

Polyumenal implantable organ

A polylumenal implantable device comprises a body defining a plurality of capillary lumina. The prosthetic device is suitable for implantation in a patient as an arterial or venous bypass graft or shunt, or intra-organ implant as well as other purposes. The improved prosthetic device has increased surface area and preferably a three-dimensional porosity for encouraging the harboring of, for example, endothelial cells, as well as for receiving organized deposition of material such as genetically enhanced cell types. A method for providing a bioactive material to a patient includes the steps of providing a polyluminal implantable organ comprising an implantable body defining a plurality of capillary lumina, treating the interior surfaces of the lumina with a bioactive material or plasma polymerization, and implanting the prosthetic device in the patient so that bodily fluids of the patient come into contact with the treated interior surfaces.

Role of endothelial cells in the development of glomerular lesions of mesangiocapillary glomerulonephritis.

The histological and ultrastructural changes of mesangiocapillary glomerulonephritis (MCGN) are not unique to this entity: splitting of basement membrane (BM) is seen in a number of conditions with an altered coagulation pattern. The distribution of endothelial cells in the glomerular capillaries in five cases of MCGN was studied by light and electron microscopy and immunocytochemistry; endothelial cells were stained with peroxidase or FITC-conjugated antibodies against Factor VIII-related antigen or CD34 antigen and observed with conventional light/immunofluorescence microscopy and confocal laser scanning microscopy for three-dimensional reconstruction. Electron microscopy was performed with colloidal gold labelling. Endothelial cells were shown to be present within the duplicated basement membranes of capillaries and continuity was demonstrated with endothelial cells lining the capillary lumina. These results suggest that endothelial cells as well as mesangial cells can participate in the formation of the characteristic double contour of capillary walls in MCGN, especially in the early stages. There are some similarities to the changes seen in larger arteries as a consequence of thrombotic disease.

Glomerular ultrastructural lesions of idiopathic cutaneous and renal glomerular vasculopathy of greyhounds.

Idiopathic cutaneous and renal glomerular vasculopathy (CRGV) (Alabama rot) is a potentially fatal disease of unknown etiology that affects the skin and kidneys of racing- and training-age Greyhounds. Ultrastructural examinations were performed on two healthy control Greyhounds and 12 Greyhounds diagnosed with CRGV based on the presence of characteristic, well-demarcated cutaneous ulcers of the extremities (12/12), thrombocytopenia (< 200,000 platelets/dl) (12/12), and acute renal insufficiency (BUN > 40 mg/dl, serum creatinine > 2.0 mg/dl) (7/12). Early glomerular ultrastructural changes included endothelial swelling, detachment, and necrosis; membranous whorl formation; and platelet adhesion and aggregation. Some capillaries were occluded with aggregated platelets, cellular fragments, and fibrin. Later changes included narrowing of capillary lumina and thickening of glomerular capillary walls by subendothelial accumulation of flocculent, amorphous, variable electron-dense material and occasionally erythrocytes, cellular processes, and fibrin. Glomerular endothelial cells were increased in number and plump, with villouslike cytoplasmic projections. Mesangial cell cytoplasmic processes occasionally were interposed between the endothelium and the basement membrane. No etiologic agents or electron-dense deposits typical of immune complexes were observed. Although the specific etiology was not determined, the ultrastructural changes suggest that glomerular endothelial damage is an important early event in the pathogenesis of CRGV.

Dermal angiopathy in hemodialysis patients: The effect of time

We studied skin biopsies from 14 patients after 6 months to 18 years on hemodialysis (HD) to discern any effect of duration of treatment on skin and dermal capillaries. Patients selected for biopsy were without evidence of other diseases known to affect vasculature such as diabetes mellitus. Pathological changes compared with duration of HD were: capillary wall thickening, endothelial proliferation and new capillary formation, lipid content, and epidermal atrophy. Severity of morphologic changes were graded from 0 to 4+ by a pathologist who had no knowledge of HD duration. The earliest change observed was reduplication of the capillary basement membrane. Narrowing of capillary lumina due to endothelial cell proliferation and new capillary formation were noted after five years of HD; lipid droplets were noted in capillaries by five years; and epidermal atrophy by 10 years. Progressive severity of each finding was associated with length of time on HD. Neither amyloid nor Ca++ deposits were observed in any specimens. By clinical observation, easy bruisability and increased skin fragility were worse the longer the patient was on HD. Capillary occlusion inducing tissue ischemia could be a cause of the atrophic skin changes noted. However, no patient manifested dermal necrosis. While pathogenesis of the capillary changes in uremic skin is unknown, the changes have been shown to stabilize following successful kidney transplantation.

Endocapillary glomerulitis in the renal allograft.

Endocapillary glomerulitis is characterized by an increase in number of mononuclear cells in the glomerular capillary lumina. This lesion has been described in the early posttransplant period, but its pathogenesis, relation to conventional rejection, and prognostic impact is not well known. Using the definitions, scorings, and gradings of the Banff system for classification and grading of histopathologic changes in the renal allograft, we have analyzed 444 consecutive renal allograft biopsies from the first 90 days posttransplant. Moderate or severe glomerulitis occurred in 13.5% of the biopsies. There was a strong tendency toward clustering of glomerulitis: if one biopsy from a patient had glomerulitis, there was a high probability that it occurred in other biopsies from the same patient. There was some correlation with conventional acute rejection, but 40% of all biopsies with glomerulitis had no rejection and 53% of all biopsies with rejection had no glomerulitis. Graft function at biopsy was nil or decreased in many patients, but this could largely be explained by the independent presence of primary graft dysfunction or conventional rejection, these conditions being a frequent indication for performing a graft biopsy. Moderate or even severe glomerulitis was, however, compatible with a functioning graft. No correlation between glomerulitis and active CMV infection was found. The one-year graft survival of grafts with early posttransplant glomerulitis was 66%. If early conventional acute rejection is taken into consideration, graft survival does not seem to be influenced by the presence of glomerulitis. Early posttransplant endocapillary glomerulitis may be a peculiar pattern of rejection with a pathogenesis different from that of conventional rejection, but the present investigation does not demonstrate any adverse effects on graft function or graft prognosis.

Ecto-5?-nucleotidase is expressed by pericytes and fibroblasts in the rat heart

Ecto-5'-nucleotidase is anchored at the outer surface of cell membranes and thus its reaction product adenosine is released into the extracellular space. Extracellular adenosine displays via specific receptors a wide range of physiological effects in heart. There are discrepancies in the literature concerning the distribution of ecto-5'-nucleotidase in heart. Since we suspected that these may be due to technical problems, in the present study on ecto-5'-nucleotidase in rat heart we attempted to circumvent some technical pitfalls. Good preservation of the tissue with open capillary lumina, providing a clear identification of endothelium, was obtained by perfusion fixation. At the light microscopic level, the distribution of ecto-5'-nucleotidase studied by enzyme histochemistry and immunohistochemistry using a monoclonal and a polyclonal antibody yielded congruent results. The enzyme was rather homogeneously distributed throughout the myocardium, with a slightly higher incidence of stained cells in the outer thirds than in the inner third of the wall. Consistently high levels of ecto-5'-nucleotidase were seen only in interstitial cells. The walls of large vessels and heart muscle cells were constantly negative for ecto-5'-nucleotidase. The endothelia of capillaries were mostly negative but a few profiles occasionally displayed a weak immunoreaction. The interstitial cells staining positive for ecto-5'-nucleotidase could be identified as pericytes and as fibroblasts according to their shapes and localizations. The immunoreactivity of fibroblasts was confirmed by electron microscopy. These data indicate that adenosine may be formed extracellularly in the interstitium of the myocardium, where it would have direct access to important targets such as myocytes, arterioles and nerve endings.

Lipoprotein glomerulopathy: A new aspect of lipid induced glomerular injury

Summary: Lipoprotein glomerulopathy is a new type of glomerular disease which is thought to be induced by an abnormality of lipoprotein metabolism. Until the end of 1994, 21 cases had been reported from Japan. However, this disease is not restricted to Japan because two cases with similar features have been described in France and the United States recently. Clinically, the presenting feature in all patients is proteinuria, resulting in a nephrotic syndrome in the majority. Renal biopsy specimens reveal that capillary lumina in the glomerulus are markedly dilated with pale‐stained and mesh‐like substances which are composed of fine granules. Under electron microscopy, it is observed that a combination of granules forms strata as fingerprints. Sudan or oil red‐O staining and immunofluorescence study on snap‐frozen sections show lipid droplets and apolipoproteins (apos) B and E in the capillary lumina, respectively. Accordingly, the intracapillary substances are thought to be lipoprotein thrombi. Lipid and lipoprotein profiles show type III hyperlipoproteinaemia and high level of apo E in plasma. However, the apo E phenotype is usually the heterozygous E2/3 or E2/4, different from the homozygous E2/2 in familial type III hyperlipoproteinaemia. Systemic manifestations characteristic of the previously‐reported lipidoses are not observed.Occurrence in several families and recurrence in two transplanted kidneys suggest that the disease may be induced by the hereditary abnormalities of lipoprotein metabolism. On the other hand, it is likely that in situ mechanisms may mediate development of the disease in the glomerulus.

Enzymatic digestion increases permeability of the outer blood-retinal barrier for high-molecular-weight substances

The purpose of the study was to investigate whether lysosomal enzymes can participate in damaging the outer blood-retinal barrier and to examine the role of glycosaminoglycans in maintaining the barrier function for high-molecular-weight substances. The ciliary artery was cannulated in freshly enucleated pig eyes. Perfusion was performed with buffer (controls), with heparinase (substrate: heparan sulfate), or with lysosomal enzymes freshly prepared from pig retinal pigment epithelium at 36 degrees C, followed by perfusion with the tracer native ferritin (NF) or the marker cationized ferritin (CF). The eyes were examined by electron microscopy. In controls treated with buffer alone, NF was found in high concentration in the lumina of the choroidal capillaries; however, little NF was found in Bruch's membrane (BsM). The tracer did not penetrate to any extent beyond BsM. In eyes digested with heparinase or lysosomal enzymes, significantly higher numbers of tracer molecules were found in BsM. Furthermore, NF penetrated BsM and was apparent in the subretinal space and also inside retinal pigment epithelial cells, probably due to pinocytosis. The results indicate that heparan sulfate proteoglycan is important for the maintenance of the outer blood-retinal barrier and that lysosomal proteases may participate in damaging this barrier, causing increased permeability to high-molecular-weight substances.

Mechanisms associated with corpus luteum development.

The transition of a preovulatory follicle into a corpus luteum is a complex process involving mechanisms similar to wound healing and tumor formation. The objective of this review is to focus on mechanisms associated with corpus luteum development with specific attention to the follicular lineage of luteal cells, mechanisms associated with luteinization, and neovascular changes during luteal development. Corpora lutea are a continuation of follicular maturation and form from granulosal and theca interna cells. There is morphological and immunological evidence in ruminant species for the differentiation of granulosal and theca interna cells into large and small steroidogenic luteal cells, respectively. Different morphological, physiological, and biochemical characteristics of large and small luteal cells may reflect different follicular lineages with separate embryological origins. Following the preovulatory gonadotropin surge, follicular cells begin morphological, endocrinological, and biochemical changes associated with luteinization. Luteinization involves the transition of a preovulatory follicle into a highly vascular corpus luteum capable of secreting large quantities of progesterone. In addition, various cell types undergo hyperplasia, hypertrophy, and(or) migration during corpus luteum formation. An essential component of corpus luteum development is the recruitment of a blood supply. The development of a new microcirculatory bed involves breakdown of the follicular basement membrane, endothelial cell migration, endothelial cell proliferation, and development of capillary lumina. This process is regulated by the interaction of angiogenic and antiangiogenic substances. Further clarification of the preceding mechanisms may result in the development of improved methodologies for controlling the time of ovulation and(or) increasing pregnancy rates.

Human glomerular growth during childhood: a morphometric study.

Morphometric measurements of glomerular size were made on renal biopsy specimens obtained from 16 children with minimal change nephrotic syndrome (MCNS) and 16 with isolated recurrent haematuria (RH), whose ages ranged from 1.8 to 15.2 years. Glomeruli were normal on light and electron microscopy, and immunofluorescence was essentially negative. Tracings were made of the outlines of glomerular capsules, tufts, and individual capillary lumens; using computerized digitometry, the mean areas of each item were determined. The number of capillary lumens per glomerulus was counted automatically. From these data, the mean mesangial area was also calculated. Mesangial cells were counted in at least 100 distal mesangial areas per biopsy. The mean glomerular capsular area (GCA) correlated well with the mean glomerular tuft area (GTA) but frequent capsular artefacts rendered the former measurement less suitable for comparison with diseased glomeruli. The mean GTA correlated significantly with age and body surface area in both MCNS and RH, as did the number of capillary lumens per glomerulus, but the mean capillary lumina area showed no such correlations. The mean mesangial area increased with age in proportion to GTA, whereas mesangial cell counts remained normal. These findings indicate that glomerular growth during childhood is proportional to body growth, and are consistent with capillary subdivision as the mechanism of enlargement.

Kidney in Galloway-Mowat syndrome: Clinical spectrum with description of pathology

The Galloway-Mowat syndrome, a rare inherited disorder, is characterized by congenital microcephaly with hypotonia and developmental delay, often hiatus hernia, and nephrotic syndrome manifested in infancy or in early childhood. The glomerular lesion has been poorly characterized in the few previous reports of this syndrome. We studied three siblings with microcephaly and nephrotic syndrome occurring during the first two weeks of life. Hematuria, glycosuria and renal failure were also present. Renal biopsy and postmortem specimens of two patients were studied. Glomerular structure was disorganized; capillary lumina were of varying calibers, capillary walls were adherent to one another, and mesangial zones were poorly demarcated. Glomerular basement membrane ultrastructure was markedly altered. The normal trilaminar structure was obscured or replaced by flocculent material; furthermore, 6 to 8 nm fibrils of unknown nature permeated the space between endothelial and epithelial cells. Non-glomerular basement membranes were unaltered in appearance. This syndrome apparently represents, in part, a new disorder of glomerular basement membrane formation and function.

Pathogenetic Mechanisms Involved in Mesangial Interposition in IgA Nephropathy

To investigate the pathogenetic mechanisms of mesangial interposition (MI) in IgA nephropathy, we examined renal biopsy samples from 20 patients with IgA nephropathy. Electron microscopic morphometric analysis showed that cytoplasmic protrusion of mesangial cells (MCs) into endothelial cells (ECs) or capillary lumina, and widening of the lamina rara interna (LRI) were significantly more prominent in glomeruli with MI than in those without. Immunoelectron microscopy revealed that the ratio of positive endothelial staining with a polyclonal antibody against human platelet-derived growth factor (PDGF) BB was significantly higher in capillary loops with MI than in those without. Enhanced capillary staining of fibronectin related to MI was not recognized. These results suggest that MI in IgA nephropathy is caused by factors such as enhancement of cytoplasmic extensibility of the MCs, widening of the LRI and chemotactic influence of PDGF-BB located in the glomerular ECs.

Renal impairment and intraglomerular mononuclear phagocytes in cholesterol-fed rabbits.

Morphological and immunohistochemical studies using the anti-macrophage monoclonal antibody RbM2 were performed in cholesterol-fed rabbits. From the beginning of the experiment, the levels of lipoproteins showed the pattern of familial type III hyperlipoproteinemia patients, and glomerular endothelial and mesangial cells had lipid deposits. By the 3rd week, RbM2-positive cells appeared in the capillary lumina. These cells became larger and increased in number after the 8th week. Although they had become very large and had increased noticeably by the 20th week, few sclerotic glomeruli were observed. We could hardly detect any proliferation of or foamy changes in the mesangial cells, and monocyte/macrophages showed no proliferative capacity within the glomeruli. These findings suggest that hyperlipidemia alone did not cause proliferation or foam transformation of the mesangial cells. The glomerular macrophages, probably derived from circulating monocytes, did not induce a glomerular injury under the short-term hyperlipidemic conditions of these experiments.

Sequential Evaluation of Radiation-Induced Glomerular Ultrastructural Changes in the Pig Kidney

Both kidneys of 12 mature female pigs received either a single dose of 9.8 Gy 60Co gamma rays or sham irradiation. At intervals of 1-4 weeks serial renal biopsies were obtained, followed by sacrifice at 24 weeks after irradiation. Individual kidney glomerular filtration rate (GFR), effective renal plasma flow (ERPF), and the hematocrit (Hct) were measured routinely. Renal irradiation resulted in a progressive decline in GFR, ERPF, and Hct, with minimal values being observed within 12 weeks of irradiation. No change in any of these parameters was noted in the sham-irradiated pigs. The initial morphological change in irradiated glomeruli was leukocyte attachment to capillary endothelial cells 3-6 weeks after irradiation followed by activation and swelling of the endothelial cells. This was followed by pronounced increases in capillary permeability with fluid and erythrocyte, leukocyte, and platelet exudation into the subendothelial/mesangial space. This resulted in compression of glomerular capillary lumina, which occurred concomitantly with the reduction in GFR. By 12 to 15 weeks after irradiation the changes in endothelial cells were less evident. However, mesangial cells exhibited evidence of activation and proliferation accompanied by progressive mesangial expansion and sclerosis. Thus the glomerular capillary endothelial and mesangial cells appear particularly important in the pathogenesis of radiation nephropathy.

Association of immunolocalization of matrix metalloproteinase 1 with ovulation in hCG-treated rabbit ovary.

Matrix metalloproteinase 1, MMP-1, which was previously called interstitial collagenase, is necessary for extracellular matrix reconstruction. The immunolocalization of the latent form of MMP-1 (proMMP-1) was examined in the ovulatory process in hCG-treated (100 iu per animal) rabbit ovaries. Immunoreactive products of proMMP-1 were identified by the avidin-biotin peroxidase complex formation using an anti-rabbit proMMP-1 polyclonal antibody. ProMMP-1 was distributed in the cytoplasm of theca interna cells, theca externa cells, interstitial glands and germinal epithelium throughout ovulation. However, at 9 or 10 h after hCG treatment, this enzyme was identified in several capillary lumina around the apex of preovulatory follicles. In addition, the staining density of immunoreactive products apparently increased in granulosa cells and theca interna cells around the orifice of the ruptured follicle 10 h after the stimulation. These results indicate that the spatiotemporal appearance of proMMP-1 in ovulation may be closely associated with the initiation of rupture of the follicle.

Co-localization of erythropoietin mRNA and ecto-5'-nucleotidase immunoreactivity in peritubular cells of rat renal cortex indicates that fibroblasts produce erythropoietin.

In adults, the kidneys are the major site of production of the glycoprotein hormone erythropoietin (EPO), but the type of renal cell producing EPO has not yet been identified. In the present study we used non-radioactive in situ hybridization with a digoxigenin-labeled cRNA probe to localize cells that produce erythropoietin (EPO) in kidneys of anemic rats. Cryostat sections from both native and perfusion-fixed tissue were used. Cells containing EPO mRNA were found exclusively in the peritubular space of the renal cortex. Using high-resolution interference contrast optics, we found that cells expressing EPO mRNA were not associated with the lumina of peritubular capillaries but rather were located in the angles between adjacent tubules or between tubules and vessels. These spaces are predominantly occupied by resident interstitial fibroblasts and by their cytoplasmic processes. To further identify the type of cell containing EPO mRNA, a double-labeling protocol was established that permitted on the same tissue section both in situ hybridization for EPO mRNA and parallel immunolabeling of ecto-5'-nucleotidase (5'-Nu), a surface marker of peritubular interstitial fibroblasts. The combined labeling technique revealed that a clear majority of cells expressing EPO mRNA also displayed staining for anti-5'-Nu. Staining for EPO mRNA was localized in central perinuclear parts of the interstitial cells, whereas 5'-Nu label was present on the cell surface, including the cytoplasmic processes. These data indicate that peritubular fibroblasts are cellular sites for production of erythropoietin.

Characteristic features of the ultrastructure of choroid plexus capillaries of fetal lateral ventricles and their clinical significance for intraventricular hemorrhage

AbstractThe capillaries within the lateral ventricular choroid plexuses from 14 normal fetal brains and one 5‐year‐old child were studied with electron microscopy. Endothelial pores and simple tight junctions were not observed until a fetal age of 24 weeks. From 24 weeks through 30 weeks of fetal age, the protein particles of the tight junctions became progressively aligned into a continuous row. Only an intermittent basement‐membranelike material was found in the fetal tissues; however, a continuous basement membrane was found in tissues from a 5‐year‐old child. We suggest fluid in capillary lumina may enter the tissue space through “very leaky” endothelial tight junctions due to the fixed osmotic gradient, while the macromolecular substances pass into the tissue space by pinocytosis of the endothelial cells. The macromolecular substances may in turn be transferred into the ventricular cavities by epithelial cells. Although the pathogeny of fetal interventricular hemorrhage (IVH) is unclear, we suspect that the simple tight junctions found early in development may break down under certain abnormal vascular conditions and allow blood to leak out. Normal deficiencies in the fetal basement membrane and its weak extensibility are also factors which could contribute to fetal IVH. No relationship was demonstrated between the types and numbers of cell organelles present and fetal age. © 1993 Wiley‐Liss, Inc.

Proliferation, migration and platelet release by megakaryocytes in long-term bone marrow culture in collagen gel.

The proliferation of megakaryocytes, their migration, and their platelet release processes were observed in long-term bone marrow culture in collagen gel. Megakaryocytes proliferated for more than 6 months, not only with myeloid cells but also with sinusoid-like capillaries. The megakaryocyte count decreased at 2 weeks of culture, increased to more than 400 at 6 weeks, and then decreased to about 100. Megakaryocyte colonies appeared after 2 weeks of culture; the number increased to more than 10 at 4 weeks of culture, and was maintained at that level. Morphologically, most fully mature megakaryocytes extended several long antennae-like processes, with periodic constrictions, demarcation membranes (DM), and platelet fields. Pro-platelets were released from these processes primarily by a "pinching off" mechanism, but some megakaryocytes released pro-platelets by the dissociation of DM, without extending long processes. Mature megakaryocytes migrated like ameba in collagen gel, and a few migrated to the abluminal side of the capillary or into the capillary; in both cases releasing pro-platelets into the capillary lumina. These observations were very similar to those noted in vivo.

Fine structure of adrenal cortex in rats harbouring a medullary thyroid carcinoma transfected with a corticotrophin-releasing hormone cDNA expression vector

We report the light microscopic, transmission and scanning electron microscopic features of the adrenal cortices in rats bearing a medullary thyroid carcinoma cell line transfected with a corticotrophin-releasing hormone (CRH) cDNA expression vector. The animals had elevated CRH, ACTH and corticosterone blood levels, involuted thymuses and markedly enlarged adrenal glands with prominent lipid-depleted cortices and dilated congested capillaries, similar to those of animals treated with ACTH. Using electron microscopy it was found that the enlarged fasciculata and reticularis zones were composed of large, compact cells with abundant smooth endoplasmic reticulum, prominent Golgi complexes, increased number of large mitochondria with focal loss of cristae and cavitation of the internal compartments, numerous lysosomes and prominent elongated microvilli. In addition, small cytoplasmic fragments were seen within the capillary lumina; these structures resembled microvilli that were apparently detached from adrenocortical cells and entered the blood stream via discontinuous endothelium of dilated capillaries. By scanning electron microscopy it was found that the cells had bulging surfaces with scattered pits and numerous long microvilli pointing in different directions. This animal model allows analysis of the effects of protracted CRH excess resembling tumoural CRH-dependent Cushing's syndrome in human patients. Our findings call attention to the role of microvilli in adrenocortical secretion. The increased number and size of microvilli has been thought to lead to an increase in the surface area of adrenocortical cells, thereby facilitating hormone discharge. The detachment of microvilli from adrenocortical cells may represent a form of apocrine secretion and may contribute to hypercorticosteronaemia in CRH excess.