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Biomedical subjects

M M Magalhães

Publications and source records attributed to M M Magalhães.

At least 19 recordsLinked to original sources
PubMed2000

Poly-A-rich domains in adrenal cell nuclear matrix after acute and chronic ACTH stimulation: an in situ hybridization study.

The regulatory effects of adrenocorticotropin (ACTH) at the adrenal cell nucleus are not yet completely clarified. Recent studies showed that eukaryotic cell nucleus is composed of specific subcompartments organized by the nucleoskeleton-nuclear matrix. In this paper, the RNA polymerase II transcriptional domains were detected in the adrenal nucleus after acute and chronic ACTH stimulation by using in situ hybridization. Polydeoxythymidine (poly-T) probe, complementary to the polyadenylated (poly-A) chain of all the mRNAs and its precursors, was used. Hybridization was performed in intact adrenal cells and in nuclear matrices isolated from previously dispersed cells. The transcriptional domains, enriched in poly-A RNA, were evident in intact adrenal cells, the staining intensity being higher both in the acutely and in the chronically stimulated animals. In the nuclear matrices poly-A-rich areas were still visible, and were more intensely stained after ACTH stimulation.

Adrenal Glands↗
PubMed1999

Serum levels of cytokines in patients envenomed by Tityus serrulatus scorpion sting.

Seventeen patients stung by Tityus serrulatus scorpion were classified as mild (pain at the site of the sting, n = 6), moderate (local pain and one of the following manifestations: vomiting, psychomotor agitation, prostration, sweating, tachypnea, tachycardia and mild arterial hypertension, n = 10) and severe cases (equal moderate cases plus cardiac failure, pulmonary edema and shock, n = 1). Venous blood was sampled for biochemical and hematological analysis and for IL-1alpha, IL-6, IL-10, TNF-alpha, IFN-gamma and GM-CSF ELISAs at the time of hospital admission, 6 h (moderate and severe cases), and 12, 18, 36 and 72 h (severe case) later. Ten age-matched healthy volunteers were used as control. Increased serum levels of IL-1alpha was noticed in all patients, high levels of IL-6, IFN-gamma and GM-CSF were observed only in a patient with severe envenomation. Our data suggest that a systemic inflammatory response-like syndrome is triggered during severe envenomation caused by T. serrulatus sting and that release of cytokines may be involved in this response.

Blood Cell Count↗
PubMed1999

Serum leptin and body composition in children with familial GH deficiency (GHD) due to a mutation in the growth hormone-releasing hormone (GHRH) receptor.

OBJECTIVE: The relationship between GH, body composition and leptin in children remains ill-defined. We have therefore examined the impact of severe GH deficiency (GHD) due to a mutation in the GHRH receptor on serum leptin concentrations and body composition in childhood. PATIENTS: 12 affected children and young people (GHD) (4 M:8F, age 5.4-20.1 years, 8 Tanner stage (TS) 1-2, 4 TS 3-5) and 40 healthy controls (C) from the same region (13 M:27F, age 5.3-18.4 years, 20 TS 1-2, 20 TS 3-5). METHODS: Percent body fat was determined by infra-red interactance, from which the amounts of fat mass (FM, kg) and fat free mass (FFM, kg) were derived. Serum leptin concentrations were measured in a single fasted, morning serum sample and results expressed as a concentration and as leptin per unit fat mass (L/FM, ng/ml/kg). To control for differences in sex and pubertal maturation, leptin standard deviation scores (leptin SDS) were calculated using normative data from UK children. RESULTS: FFM was significantly lower in GHD children than in controls (TS 1-2 P < 0.05, TS 3-5 P < 0.001). FM did not differ significantly between the two groups. Serum leptin concentrations, leptin per unit fat mass and leptin SDS were significantly elevated in GHD children both peripubertal and pubertal compared with controls. Using all subjects, stepwise multiple linear regression with FM, FFM, age, puberty and sex as explanatory variables and leptin concentration as the dependent variable indicated that 59% of the variability in leptin could be accounted for by FM (+, 45%), FFM (-, 9%) and sex (+, 5%) (P < 0.001). However on inclusion of GH deficiency (coded GHD = 1, control = 2) as an explanatory variable 73% of the variability in leptin was explained by FM (+, 45%), GHD (-, 22%) and sex (+, 6%) (P < 0.001). CONCLUSIONS: These data indicate that severe GH deficiency in children is associated with elevated leptin concentrations, irrespective of sex or pubertal stage. This increase is not associated with differences in fat mass but is related to reduced fat free mass in GH deficiency. Furthermore in this population there may be an additional effect of GH deficiency on leptin, independent of the influences of sex and body composition.

Adolescent↗
PubMed1999

Kinetics of neovascularisation of splenic autotransplants in mice.

The aim of this study was to examine the kinetics of neovascularisation of splenic autoimplants into the abdominal cavity after splenectomy in mice. Sixty-eight female Swiss mice were submitted to splenectomy. The spleen from each animal was sliced and the slices were implanted into the abdominal cavity. Groups of animals were killed after 1, 2, 3, 5, 7, 15, 21, 28, 42, 56, 70 and 84 d. Fluorescent polystyrene microspheres were injected via the orbital venous plexus before killing and the splenules were removed 5 min later for light and electron microscopy. Mesenteric blood vessels were injected with coloured latex to study the origin of the nutrient vessels. Three days after the implant the microspheres were observed at the periphery and then migrating to the internal parts of the implant in the subsequent days. The blood supply to the implants originated from branches of the splenic, short gastric, mesenteric and gastroepiploic arteries. It is concluded that revascularisation of splenic autografts proceeds centripetally, starting as early as 3 d after implantation.

Animals↗
PubMed1999

Ontogeny of 3beta-hydroxysteroid dehydrogenase expression in the rat adrenal gland as studied by immunohistochemistry.

The enzyme 3beta-hydroxysteroid dehydrogenase plays a crucial role in the steroidogenic process in the adrenal gland. In the present study we tried to characterize its localization and developmental changes in the rat adrenal cortex during the postnatal period, using immunohistochemical methods. The development of the different zones evidenced specific particularities: the zona glomerulosa almost lacked 3beta-HSD in the first days after birth; then, 3beta-HSD increased, attaining a maximum around day 20 and afterwards it decreased again and remained less intense than the neighbouring zona fasciculata up until adulthood (65 days of age). The zona fasciculata was already intensely stained at birth and the expression of 3beta-HSD increased rapidly reaching a maximum after 2 weeks of life and that level was maintained from then on. The inner part of the zona fasciculata and the zona reticularis both of which develop postnatally were faintly immunostained before day 20. The expression of 3beta-HSD increased after that age to become approximately as intense as in the outer zona fasciculata and so remaining until day 90. The development of the zona glomerulosa was parallel to the secretion of aldosterone. The same did not occur with the zona fasciculata as the intensity of staining during the first 14 postnatal days was accompanied by very low levels of corticosterone.

3-Hydroxysteroid Dehydrogenases↗
PubMed1999

Effects of prolonged infusion of basic fibroblast growth factor and IGF-I on adrenocortical differentiation in the autotransplanted adrenal: an immunohistochemical study.

Adrenocortical regeneration after adrenal autotransplantation provides a model for the study of local autocrine/paracrine mechanisms involved in the growth and differentiation of the adrenal cortex. To study the possible involvement of some growth factors, namely basic fibroblast growth factor (bFGF, FGF-2) and insulin-like growth factor I (IGF-I), in cell differentiation, immunohistochemical and ultrastructural studies were carried out on adrenal autotransplants in adult male rats. To distinguish between fasciculata and glomerulosa-like cells with accuracy, tissue sections were immunostained with IZAb, which recognizes the inner zone antigen (IZAg) present in fasciculata and reticularis cells but absent from the glomerulosa, and by electron microscopy. IGF-I-treated animals exhibited a clear glomerulosa-like zone that was devoid of IZAb immunostaining. In this outer subcapsular area, ultrastructural examination showed cells containing mitochondria with irregular cristae resembling those of the fetal or immature glomerulosa cells. In contrast, no significant morphological differences were observed in bFGF-treated animals when compared with those from saline-treated controls, in both of which, IZAb immunostaining occurred in almost all adrenocortical cells, with no clear zonation or glomerulosa, as seen in the intact animal. Plasma aldosterone and corticosterone concentrations were lower in autotransplanted control animals than in intact controls, although plasma renin activities were similar. IGF-I treatment significantly increased aldosterone concentrations, whereas corticosterone and plasma renin activity were reduced. bFGF infusion further reduced plasma aldosterone, although plasma renin activity and corticosterone were unaffected. These results suggest that the two growth factors have different effects on zonal differentiation and function in the autotransplanted gland. In particular, bFGF, by reducing glomerulosa function, appears partly to replicate the actions of ACTH in normal animals. In contrast, IGF-I enhances the glomerulosa secreting phenotype and diminishes that of the fasciculata/reticularis, possibly replicating the actions of angiotensin II or a low sodium diet.

Adrenal Cortex↗
PubMed1999

Immunocytochemical detection of structural and regulatory proteins in rat adrenal nuclear matrix.

The nuclear matrix is a specific cell structure consisting of a residual nucleoskeleton that extends from the nucleoli to the nuclear envelope. The nuclear matrix of steroidogenic cells was isolated previously from a purified nuclear fraction. We present here an in situ extraction method, modified Lutz's method, for rat glandular adrenal cell nuclear matrix. This residual organelle was characterized and studied using immunocytochemical methods. The adrenal glands were removed, the cells prepared in suspension and deposited by cytospin onto Poly-L-lysine glass slides. The nuclear matrix was extracted with Nonidet P-40, DNase I and high and low ionic strength buffers. Structural proteins, nuclear lamins, coilin and fibrillarin were detected immunocytochemically. The adrenal fasciculata cells were easily identified by this method because of their large nuclei and abundant lipid droplets in the cytoplasm. After immunocytochemical detection by antibodies against lamins A and C, a marked brown layer at the periphery of the nucleus was observed. The intensity of the staining was lower using the antibody against nuclear lamin B. Immunocytochemical detection of the protein coilin revealed punctuated stained areas, 2-6 per nucleus, that probably correspond to the coiled bodies. The protein fibrillarin was detected at the nucleolus and coiled bodies. Our technique is simple, reveals well preserved adrenal nuclear matrices, and may be a useful method for immunocytochemical analysis and in situ hybridization.

Adrenal Glands↗
PubMed1998

Age-related changes in the inner zone of the adrenal cortex of the rat--a morphologic and biochemical study.

In this work, a correlative morphologic and biochemical study on the effects of ageing on the rat adrenal Inner Zone (IZ) was made. Male Wistar rats were studied at 2, 6, 12, 18 and 24 months. Structural data of Zona Fasciculata (ZF) showed age-related increase in cell volume (P < 0.05), decrease in mitochondria (P < 0.01) and smooth endoplasmic reticulum (SER) volumes, and increase in lipid droplets (P < 0.01) and lipofuscin granules (P < 0.01) volumes. In Zona Reticularis, the main change observed was the increase in lipofuscin granules (P < 0.001). Serum corticosterone from unstimulated rats increased until 12 months but decreased thereafter (P < 0.01), to levels below those from 2-month-old rats. Similarly, plasma adrenocorticotrophic hormone (ACTH) presented a maximum at 12 months, followed by a decrease to levels higher than at 2 months (P < 0.05). In rats injected either with only ACTH or dexamethasone, before ACTH stimulation, corticosterone level had a maximum at 12 months. In aged rats, serum high density lipoprotein (HDL) and adrenal cholesterol ester increased significantly (P < 0.05 and P < 0.01, respectively), whereas adrenal corticosterone decreased. Products of lipid peroxidation, assayed with the thiobarbituric acid reaction and fluorimetry showed an age-related increase (P < 0.05). The age-related decrease in mitochondria and SER volumes is consistent with the decrease of serum corticosterone. The increase in lipid droplet and HDL and the reduction of adrenal corticosterone level correlate with the increase of adrenal cholesterol ester content. These suggest a continued uptake of steroid precursor but a reduced steroid synthesis. On the whole, the data provide evidence for an age-related reduced functional ability of IZ and particularly of ZF.

Adrenal Cortex↗
PubMed1998

The development of the adrenal gland zona glomerulosa in the rat. A morphological, immunohistochemical and biochemical study.

We have studied the development of the adrenal gland in the rat comprising the ages ranging from 0 to 90 days after birth. The weight of the animals and that of the adrenal glands demonstrated a linear growth with time until 75 days, both in males and females. The area of the zona glomerulosa (ZG) increased in size from birth until approximately 40 days of age. After that, growth had a much smaller slope (females, r=0.84, P < 0.001; males, r=0.81, P < 0.001). Aldosterone secretion had a marked increase until 20 days of age and thereafter demonstrated a tendency for a decrease (females, r=-0.19, P < 0.02: males r=-0.26, P < 0.001). Plasma renin activity followed a trend parallel to that of aldosterone. The steroid precursor 18-OH-deoxycorticosterone (18-OH-DOC) demonstrated a different course as it increased progressively with age especially in the females (females, r=0.57, P < 0.001; males, r=0.40, P <0.001). The expression of the enzyme 3-beta-hydroxysteroid dehydrogenase (3-beta-HSD) was also studied by immunohistochemistry and it was shown to be very low at birth and starting to increase by 10 days of age. After 30/40 days of age the amount of this enzyme existing in the ZG was comparable with that of the outer zona fasciculata (ZF). We conclude that the development of the ZG in the rat has particularities that make it different from that of the rest of the cortex.

3-Hydroxysteroid Dehydrogenases↗
PubMed1997

Peroxisomes in adrenal steroidogenesis.

Peroxisomes, cytoplasmic organelles limited by a single membrane and with a matrix of moderate electron density, are present in a great number of cells, namely in adrenal cortex and other steroid-secreting organs. Presently peroxisomes are considered to be involved in important metabolic processes. They intervene in: (1) the production and degradation of H2O2; (2) biosynthesis of ether-phospholipids, cholesterol, dolichol, and bile acids; (3) oxidation of very long chain fatty acids, purines, polyamines, and prostaglandins; (4) catabolism of pipecolic, phythanic and glyoxylic acids; and (5) gluconeogenesis. Recent studies demonstrated that the experimental alterations in the normal steroidogenesis, produce significant morphological and biochemical changes in peroxisomes. Besides this, the presence of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (the key enzyme in the de novo cholesterol synthesis from acetate) and of sterol carrier protein-2 (SCP2), which is involved in the cholesterol metabolism and steroid metabolic pathways, are located in peroxisomes of steroid-secreting cells. In addition, patients with peroxisome diseases present deficiency in steroidogenesis, as well as reduced levels of SCP2. These data pointed out the important role of peroxisomes in steroid biosynthesis.

Adrenal Cortex Hormones↗
PubMed1996

Modulation of autotransplanted adrenal gland by endothelin-1: a morphological and biochemical study.

BACKGROUND: Adrenal gland autotransplantation, a model of cortical tissue regeneration, provides the reconstruction of distinct functional and morphological zonae. A morphological and biochemical study of the adrenal gland of adult male rats after autotransplantation and endothelin-1 (ET-1) administration was made. METHODS: The technique involved bilateral adrenalectomy and placement of pieces of the adrenal gland in a dorsal plane between the skin and muscle. The animals were killed 90 days after the autotransplantation and 1 hr after intravenous ET-1 administration (0.5 microgram/kg body weight). The autotransplanted pieces were removed, fixed, and processed for light and electron microscopic morphologic studies. Trunk blood was collected for steroid assay. RESULTS: Saline-treated control autotransplanted animals showed no remarkable differences in adrenal organization; grafts exhibiting a mass of regenerated cortical tissue were arranged in nests of glandular cells surrounded by a fibrous capsule and intersected by layers of connective tissue. The adrenal medulla was systematically absent. Ultrastructure of ET-1-treated animals revealed an inner area in the graft, consisting mainly of fasciculatalike cells. Cytoplasmic changes were evident, with high variations in mitochondrial size and arrangement. Profiles of smooth endoplasmic reticulum sometimes exhibited evidence of hypertrophy. Glandular cells in the graft outer area (subcapsular) were almost invariably like glomerulosa; however, some of them showed mitochondria with a peculiar arrangement of the cristae. "Hybrid" cells with mitochondria resembling those of the zona reticularis were also observed in the subcapsular environment. ET-1-stimulated animals showed significant increases in plasma corticosterone and aldosterone concentrations. CONCLUSIONS: Endothelin-1, previously reported to stimulate acutely the aldosterone secretion by the adrenal zona glomerulosa in the rat, seems to exert a modulator role on the physiology of adrenal autotransplants, their regeneration and secretion.

Adrenal Glands↗
PubMed1996

Acute stress effects on the adrenal cortex in the rat. A biochemical and immunohistochemical study.

Activation of the stress system induces physiologic alterations as well as behavioural ones that ultimately improve the adaptability of the organism to adverse conditions. In our previous study on the morpho-functional evolution of the adrenal cortex, from birth to adulthood, the question of what could be the contribution of immobilization stress to the observed hormonal levels was brought up. Male adult rats were submitted to immobilization of variable duration. The antibody IZAb was used to allow a correct differentiation between the zona glomerulosa (ZG) and the inner zones of the cortex (IZ). A significant increase of the ACTH levels, especially at 5 and 30 min was observed. Corticosterone (B), surprisingly, revealed 2 peaks of secretion: one at 30 sec and another at 30 min. The area of the cortex, determined by an image analyser, only showed a slight decrease at 30 sec. The proportions of the cortical area occupied by ZG and IZ were unaltered. We concluded that a corticosterone peak at 30 sec precedes the elevation of ACTH induced by stress. Only the second peak, in view of its parallel course to ACTH, can be attributed to an effect of this pituitary hormone.

18-Hydroxydesoxycorticosterone↗
PubMed1996

New insights into zonal differentiation of adrenal autotransplants in the rat: an immunohistochemical study.

Adrenal gland autotransplantation, an interesting model of adrenal regeneration, provides the reconstruction of distinct functional and morphological zonae. An immunohistochemical study of the adrenal gland of adult male rats after autotransplantation and endothelin-1 (ET-1) stimulation was carried out. The technique involved total adrenalectomy and immediate autotransplantation of small adrenal pieces under the skin of the dorsal region. The animals were killed 90 days after the autotransplantation and 1 h after intravenous ET-1 administration. Sections of recovered adrenal grafts were incubated with IZAb, a monoclonal antibody which interacts with an antigen (IZAg) predominantly found in rat adrenal inner zones. Saline-treated control autotransplanted animals showed IZAb immunostaining in almost all adrenocortical tissue, with the exception of small clusters of cells beneath the capsule. ET-1-treated animals exhibited an extended zone devoid of immunostaining and located in the subcapsular area. In addition, ET-1-stimulated animals showed significant increases in aldosterone as well as corticosterone concentrations in plasma. These results revealed that ET-1 stimulated the development of an extended subcapsular zone lacking IZAg expression, an effect that suggests its role in zona glomerulosa induction in these animals.

Adrenal Glands↗
PubMed1993

Adrenal nuclear matrix isolation. A morphologic and biochemical study.

The nuclear matrix of adrenal cells was isolated by using the methods proposed by Commerford et al and Kaufmann et al for the liver nuclear matrix isolation. Both methods permitted, to the best of our knowledge for the first time, to prepare the nuclear matrix of a steroidogenic cell and therefore to study some regulatory mechanisms governing steroidogenesis. Commerford et al's method retains nuclear envelope and so produces a higher contamination; Kaufmann et al's method presents a higher purity since the nuclear envelope was removed by Triton X-100. No RNase digestion has been employed for the isolation of the residual nuclear matrices. Both methods however, permit the isolation of fractions with a good morphology, retaining a reticular nucleolus, interchromatinic granules, and a fibrogranular scaffold extending from the nucleolus to the nuclear lamina. The major peptides detected by 1-D SDS-PAGE were 123, 56, 46 and 41 kDa; with both methods protein profiles were similar. Identification of proteins by immunodetection reveals lamins A and C, 80 and 65 kDa respectively; no labeling was found for actin (45 kDa) and vimentin (57 kDa). In short, adrenal nuclear matrix was isolated, Kaufmann et al's method being the method of choice.

Adrenal Cortex↗
PubMed1992

Autotransplantation of the adrenal cortex: a morphological and autoradiographic study.

A morphological and autoradiographic study was made of the adrenal gland of adult male rats after autotransplantation. The simple technique involved placement of pieces of the adrenal gland in a dorsal plane between the skin and muscle. Animals for morphological studies were sacrificed at 2, 3, 4, 7, 15, 30, 90, and 180 days after autotransplantation. Those for autoradiographic studies were sacrificed at 2, 3, 7, and 15 days after autotransplantation, with 3H-thymidine being administered intraperitoneally 2 h before sacrifice. Sham-operated animals were used as controls. The majority of glandular adrenal cells suffered necrosis in the first days (2 and 3) after autotransplantation. Up until 15 days and after revascularization, morphological features of the cells were compatible with protein synthesis exhibiting a developed RER, scarce SER, and mitochondria with tubular and lamellar cristae. These data may correspond to a proliferative phase of glandular cells. At day 15, cells showed morphological signs of steroidogenic activity (mitochondria with vesicular cristae, increase of SER), and at day 30, an increased number of microvilli were seen. Between 30 and 90 days zonation of the adrenal was evident with glomerulosa, fasciculata, and reticular zones readily apparent. The quantitative analysis showed a significant increase of the volumetric density of mitochondria and microvilli between the days 7 and 30. Autoradiographic studies showed an intense labelling of fibroblast-like cells at days 2 and 3 and of glandular cells at days 7 and 15, which was confirmed by the quantitative studies. Corticosterone in autotransplanted animals decreased during the first 15 days, but after 30 days the values were similar to controls. The model reported here seems to be good for study of the regeneration of the adrenal gland and can be a simple, useful, and reproducible method for adrenal transplantation.

Adrenal Cortex↗
PubMed1991

Amitosis in human adrenal cells.

Adrenal pieces obtained from 3 female and 2 male patients showed morphological figures of amitosis in adrenal zona reticularis cells. Such aspects were observed in both normal and hyperactive adrenals. Nuclei appeared constricted, heavily stained, with coarse chromatin, sometimes scattered among cytoplasmic organelles, but never marginating in crescentic caps. Cleavage of the cells originated two halves with a nucleolus in each pole. Binucleated cells were also seen in zona reticularis. The meaning of amitosis in human adrenal is discussed.

Adolescent↗
PubMed1989

Nuclear bodies in the normal and hyperfunctional human adrenal cortex.

Adrenal pieces obtained from six female patients, three without increased adrenocortical function and three with Cushing's disease, showed, in all adrenal cortex zones, cells containing simple and complex nuclear bodies. The simple nuclear bodies were spherical or ovoid and had a filamentous structure surrounded by a clear halo. Complex nuclear bodies were more numerous and heterogeneous in patients with adrenal pathology, and they were spherical with a proteinaceous filamentous capsule surrounding a core; the core was granular, filamentous or a mixture of granular and filamentous material, sometimes with a reticular or concentric arrangement. Some bodies showed vacuolar or multilocular aspect, and others had a close relationship with the nucleolus or appeared near the interchromatin granules. The meaning of adrenal nuclear bodies is discussed as well as their relationship with ACTH stimulation.

Adolescent↗