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Biomedical subjects

M Marques

Publications and source records attributed to M Marques.

At least 19 recordsLinked to original sources

Distribution of glycogen phosphorylase and cytochrome oxidase in the central nervous system of the turtle Trachemys dorbigni.

Glycogen phosphorylase (GP) and cytochrome oxidase (CO) activities were mapped histochemically in the brain of the turtle Trachemys dorbigni. In the telencephalon, both activities occurred in the olfactory bulb, in all cortical areas, in the dorsal ventricular ridge, striatum, primordium hippocampi and olfactory tubercle. In the diencephalon, they were identified in some areas of the hypothalamus, and in rotundus and geniculate nuclei. Both reactions were detected in the oculomotor, trochlear, mesencephalic trigeminal nuclei, the nucleus of the posterior commissure, torus semicircularis, substantia nigra and ruber and isthmic nuclei of the mesencephalon. In all layers of the optic tectum GP activity was found, but CO only labelled the stratum griseum centrale. In the medulla oblonga both enzymes appear in the reticular, raphe and vestibular nuclei, locus coeruleus and nuclei of cranial nerves. In the cerebellum, the granular and molecular layers, and the deep cerebellar nuclei were positive for both enzymes. The Purkinje cells were only reactive for CO. In the spinal cord, motor and commissural neurones exhibited a positive reaction for the two enzymes. However, CO also occurred in the marginal nucleus and in the lateral funiculus. These results may be useful as a basis for subsequent studies on turtle brain metabolism.

Animals

Pharmacokinetics of radioiodinated growth hormones in the turtle Chrysemys dorbigni.

Growth hormone binding proteins (GHBP) have been identified in the blood of many species. The aim of the present work is to study the physiological role of the GHBP in the turtle serum which we recently described. Binding studies were carried out using in vivo pharmacokinetic and chromatographic techniques as well as in vitro methods. When (125)I-GH was injected in physiological concentration into Chrysemys dorbigni turtles, the first step of pharmacokinetics was the binding of a significant fraction of the labeled GH by the GHBPs present in serum. The decay curve followed a three compartments model and gave the equation: Ae(-alphat) + Be(-betat) + Ce(-gammat). The fast compartment with t(1/2) of 14.4 min or 25.2 min, for hGH and bGH represents 30.3% and 18.9% of total radioactivity, respectively, at hypothetical time zero (not experi mental). Chromatographic studies reveal that this rapid compartment represents free GH. The second and third compartments represent complex forms between GH and GHBPs present in the turtle serum, and represent 70% and 80% of total radioactivity for hGH and bGH, respectively. In vitro chromatographic studies showed direct evidence of the presence of GHBPs in the turtle serum. The presence of these GHBPs changed the pharmacokinetics of labeled GH in plasma and the subsequent liver uptake of GH. The labeled hGH or bGH binds to turtle serum in similar proportion, but maximal liver uptake of these hormones are completely different (L/B ratio of 9.2 +/- 0.6 (n = 5) for ( 125)I-hGH and 4.8 +/- 0.3 (n = 7) for (125)I-bGH). The reasons for these differences could be that human GH binds to lactogenic and somatotropic receptors and bovine GH binds only to somatotropic receptors.

Animals

Axotomy increases NADPH-diaphorase activity in the dorsal root ganglia and lumbar spinal cord of the turtle Trachemys dorbigni.

Seven days after transection of the sciatic nerve NADPH-diaphorase activity increased in the small and medium neurons of the dorsal root ganglia of the turtle. However, this increase was observed only in medium neurons for up to 90 days. At this time a bilateral increase of NADPH-diaphorase staining was observed in all areas and neuronal types of the dorsal horn, and in positive motoneurons in the lumbar spinal cord, ipsilateral to the lesion. A similar increase was also demonstrable in spinal glial and endothelial cells. These findings are discussed in relation to the role of nitric oxide in hyperalgesia and neuronal regeneration or degeneration.

Animals

Functional glomerular reserve in recipients of en bloc pediatric transplant kidneys.

BACKGROUND: The transplantation of an adequate renal mass is increasingly recognized to be of importance. The improved graft survival is probably due to a lesser risk of developing hyperfiltration-associated lesions. METHODS: We have reviewed the glomerular reserve in our recipients of en bloc pediatric transplant kidneys after an intravenous amino acid overload and compared them to single adult kidney transplant recipients. RESULTS: En bloc transplants evidenced increased glomerular filtration rate as compared with baseline as from the second hour of amino acid infusion (from 71+/-14 to 84.9+/-17 ml/min, 1.73 m2, P<0.05) and increased renal plasma flow as from the third hour (from 335+/-116 to 402+/-155 ml/min, 1.73 m2, P<0.05). In the single adult kidney recipient group, no change was seen either in the glomerular filtration rate (from 62.5+/-13 to 58.1+/-13 ml/min, 1.73 m2, P=NS) nor in renal plasma flow (from 354+/-125 to 304+/-98 ml/min, 1.73 m2, P=NS). CONCLUSIONS: These results show that patients receiving en bloc pediatric kidney transplantations have a greater renal functional reserve and show a lesser risk of hyperfiltration.

Adult

[Uveitis].

Uveitis is a general term that refers to the inflammation of uveal tract, which is an important cause of blindness in young people. It is well known that uveitis can be the initial manifestation of a systemic disease (S.D.), and may appear years before the diagnosis of the primary disease. Uveitis should be integrated in a systemic study with proper testing. Therefore, the diagnosis is a matter for the ophthalmologist and the Specialist in internal medicine. We have made a retrospective study of 71 patients with chronic uveitis or panuveitis. We found 54.9% of primary uveitis and 45.1% of S.D. associated uveitis, most of them with Behçet's disease (16/71) and Ankylosing Spondilytis (7/71). HLA typing of the patients showed a decreased frequency of HLA A1 and HLA A3 antigens and an increased frequency of the HLA B27 antigen, when compared to a Portuguese control population. We confirmed the important role of HLA B27 as an independent susceptibility factor for anterior uveitis. The lowest HLA A3 frequency was observed in the group of S.D. associated uveitis, which could suggest that this antigen may play a role as a factor of resistance to uveitis.

Adolescent

Insulin binding sites in gills of the estuarine crab Chasmagnathus granulata.

Bovine 125I-insulin was injected into the estuarine crab Chasmagnathus granulata in order to study its distribution and specific uptake by tissues. The highest radioactivity uptake occurred in both anterior and posterior gills, which reached maximum values at 30-60 min following labeled insulin administration. Heart and hepatopancreas concentrated a very low amount of radioactivity (only 9 and 3%, respectively, of that shown by gills). A significant reduction of the uptake was observed in the gills when an excess of unlabeled insulin was injected together with the labeled hormone. In vitro studies also showed specific uptake of 125I-insulin by the gills incubated at 25 degrees C, which reached a plateau after 120-min incubation, suggesting a saturable process. The inhibition of 125I-insulin uptake was dose dependent on unlabeled insulin. Glucagon did not compete with radioactivity uptake by gills in vivo and in vitro. Further characterization of insulin-binding sites was performed in gill membrane. The amount of unlabeled insulin that prevented 50% of the 125I-insulin uptake was 7.78 micrograms/ml, and the Scatchard plot analysis established the presence of binding site with Kd of 3.11 microM and Bmax of 0.14 microM (r = 0.99). Ovine prolactin was not able to prevent. 125I-insulin binding to gill membrane. These findings seem to indicate the presence of specific binding sites for insulin or insulin-like substance in crab gills, which deserves further studies.

Animals

Identification and initial characterization of serum growth hormone binding protein in the turtle Chrysemys dorbigni.

Proteins that bind growth hormone (GHBP) have been identified in the blood of many mammalian and avian species, but not in reptilian species. We carried out binding studies with the serum of turtles using chromatographic techniques as well as the dextran-charcoal separation method. As in other species, we found at least two different GHBPs: one with high MW and low affinity and the other with lower MW and higher affinity. The high affinity GHBP was partially purified using gel filtration and affinity chromatography, reaching a degree of purification of 11,000 times (0.17 nmol/g of serum protein in the serum vs 1900 nmol/g protein in the purified material). When the high affinity GHBP was characterized, it was found to have a dissociation constant (Kd: 2.6 +/- 0.7 nM) similar to those described for mouse or rat, but lower than those for chicken, rabbit or man. The binding capacity (Bmax) was 120 +/- 43 fmoles/mg of protein, which can be also expressed as 1.08 +/- 0.38 pmol/ml of serum. A preliminary MW estimation of 50-60 kDa was obtained for turtle higher affinity GHBP. The specificity of this high affinity GHBP is somatogenic, since bovine GH competes as well as human GH for 125I-hGH bound to binding protein, while ovine PRL competes only partially and with low affinity. Unrelated hormones, as insulin and glucagon, can not displace the 125I-hGH bound to turtle GHBP. A very important seasonal variation in turtle GHBP activity was observed: maximum binding was found in November (springtime), followed by a continuous decline over March and May.

Animals

Acromesomelic dwarfism: a new variation.

The rate skeletal disorder, acromesomelic dwarfism, is characterized by short stature and short limbs. Bone dysplasia is evident. We report two cases of a variation of this disorder in a Portuguese woman and her son. The clinical features of these two cases differ from those of cases previously reported in the literature.

Adult

Decreased basal and acute insulin-stimulated effect on the uptake of glucose and amino acid in vitro by thyroid glands from streptozotocin-diabetic rats.

Since experimental diabetes in rats and mice is associated with impairment of several aspects of thyroid function, we determined glucose and amino acid uptake in vitro by isolated thyroid glands from normal and streptozotocin-diabetic rats. Adult male Wistar rats weighing 150-200 g were used. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ, 65 mg/kg body weight) and after five days only rats with blood glucose levels higher than 250 mg/dl were used. The thyroid glands were preincubated in Krebs-Ringer bicarbonate buffer in the presence or absence of insulin (0.7 nM to 7 muM) for 90 min and then incubated with the same concentration of the hormone or its vehicle plus 0.2 microCi of [1-14C]-2-deoxy-D-glucose ([14C]DG) or [1-14C] methylaminoisobutyric acid ([14C]MeAIB) for 15 to 180 min. The uptake of [14C]DG or [14C]MeAIB by the thyroid glands of normal rats increased as a function of incubation time, and the presence of insulin (7 microM) induced a significant increase of labelled DG from 3.30 +/- 0.11 to 4.16 +/- 0.12 and of labelled meAIB from 1.79 +/- 0.06 to 3.10 +/- 0.17 tissue/medium ratio (T/M) and after 45 min of incubation. The lowest concentration of insulin that increased both [14C]DG and [14C]MeAIB transport was 7 nM. Thyroid glands from STZ rats exhibited lower basal values of [14C]DG (4.03 +/- 0.11 T/M) or [14C]MeAIB uptake (1.05 +/- 0.05 T/M) than glands from normal rats (4.62 +/- 0.13 and 1.70 +/- 0.08 T/M, respectively). Insulin produced a stimulatory effect on the transport of both substrates in STZ rats. However, the maximal stimulating concentration of the hormone did not restore [14C]DG and [14C]MeAIB uptake to control values (4.89 +/- 0.17 in STZ rats versus 5.44 +/- 0.17 T/M in controls for [14C]DG, and 1.51 +/- 0.11 in STZ rats versus 2.19 +/- 0.10 T/M in controls for [14C]MeAIB). These results indicate that insulin exerts a direct action on the thyroid gland, and its absence or reduction affects thyroid metabolism, contributing, at least in part, to the abnormality in thyroid function associated with diabetes mellitus.

Amino Acids

[Primary lymphomas of the brain not related to immunodeficiency].

Primary brain lymphoma (PBL) is a rare disease, the study of which is based on small series and case reports. The AA review new cases of PBL not associated to immunodeficiency at the Portuguese Institute of Oncology of Porto since 1978. Five male patients were found with lymphoma primarily located in the brain, with aggressive histology, IE staging, detected by CAT, NMR or brain scintigraphy and with histologic material collected by stereotaxic biopsy (3 cases) or by craniotomy (2 cases). Three patients were in complete remission (25,5 and 1 year) and 2 were dead with a survival of 12 and 17 months. The cases and literature are discussed.

Adult

Fat-tissue injection versus graft: experimental study in rabbits.

An experimental study of fat-tissue implants in the ears and frontal regions of rabbits was conducted, starting with blocks of tissue for surgical implantation or with multifragmented pieces for injection. The implants were observed from day 7 to month 12. Low integration levels were observed in all animals, except for 1 case that had 20% and 42% of integration, respectively, for the block grafts and for the injected multifragmented tissue.

Adipocytes

Immunohistochemical characterization of mononuclear cells in delayed hypersensitivity reactions to Paracoccidiodes brasiliensis (paracoccidioidin test).

The density and distribution of T cells, T helper cells, macrophages and B cells at the site of skin tests with a cytoplasmic Paracoccidioides brasiliensis antigen (paracoccidioidin) was studied at 24 and 48 h post-challenge in 10 patients with the chronic form of paracoccidioidomycosis and in 5 non-infected individuals. The in situ study was carried out using immunoperoxidase techniques and monoclonal antibodies. The controls showed negative skin test. In the patients, the great majority of the cells in the perivascular foci were T cells (CD43-positive cells) making up 47% and 48.6% of the total number of cells at 24 and 48 h respectively. Most of the T cells showed a T helper phenotype (CD45RO-positive cells). Approximately 25% of the cells were macrophages (CD68-positive cells) and there were very few B lymphocytes (CD20-positive cells). The present data on the microanatomy of paracoccidioidin skin test sites were consistent with a delayed type hypersensitivity pattern. Our results were comparable to those reported on skin tests for other granulomatous chronic diseases.

Adult

Isolation, purification and primary structure of insulin from the turtle Chrysemys dorbigni.

Insulin A and B chains from pancreas of the turtle Chrysemys dorbigni have been purified to homogeneity, and their primary structures have been determined. The sequence of the A chain is G-I-V-E-Q-C-C-H-N-T-C-S-L-Y-Q-L-E-N-Y-C-N, and that of the B chain is A-A-N-Q-H-L-C-G-S-H-L-V-E-A-L-Y-L-V-C-G-E-R-G-F-F-Y-S-P-K-A. The amino acid sequence of Chrysemys insulin is identical to that of another turtle (Pseudemys scripta), the chicken, and turkey. When compared with alligator insulin, it has three conservative substitutions in the B chain. However, there are seven substitutions when compared with the insulin of the rattlesnake.

Amino Acid Sequence

In vitro effect of insulin on the uptake of glucose and alpha-aminoisobutyric acid in the thyroid gland of the turtle (Chrysemys dorbigni).

The presence of specific insulin binding sites in the thyroid gland of the turtle Chrysemys dorbigni has been previously reported. The purpose of the present work was to investigate the probable action of insulin on the uptake of [14C]deoxy-D-glucose ([14C]DG) and [14C]alpha-aminoisobutyric acid ([14C]AIB) in turtle (C. dorbigni) thyroid glands in vitro. Thyroid fragments (+/- 40 mg) were incubated at 25 degrees in Krebs-Ringer-bicarbonate buffer containing 0.2 microCi of [14C]DG or [14C]AIB without or with bovine insulin at different periods of time. The uptake of [14C]DG and [14C]AIB increased with incubation time. The presence of insulin (7 x 10(-6) M) in the incubation medium during 240 min did not modify the [14C]DG uptake. However if the thyroid fragments were previously incubated with insulin (60 min) and then incubated (240 min) with the same concentration of the hormone, the [14C]DG uptake was markedly increased. This stimulatory effect of insulin was dose-dependent. In similar experimental conditions, insulin also produced a significant increase in the uptake of [14C]AIB. Therefore, these findings strongly support the hypothesis that insulin might exert a direct action on the thyroid function.

Aminoisobutyric Acids

Immunohistochemical localization of insulin-like materials in antral gastric mucosa and intestinal epithelial cells of the turtles Chrysemys dorbigni and Phrynops hilarii.

Immunoreactive insulin was demonstrated immunohistochemically with antibodies to human and porcine insulin by the avidin-biotin-peroxidase complex method in open-type gastrointestinal cells from sections of the antral stomach and of the upper, middle and lower intestine of the turtles Chrysemys dorbigni and Phrynops hilarii. In both species the concentration of cells positive for insulin-like material was higher in the gastric antrum than in the gut. The localization of insulin-like material in gastrointestinal mucosal cells of turtles is an unusual finding among vertebrates, because the insulin-containing cells migrate from the mucosal epithelium of the intestine early in vertebrate evolution to the acinar pancreas. The chemical nature of the gastrointestinal insulin-like material and its physiological role remain to be determined.

Animals

Glycemia and immunohistochemical changes in the endocrine pancreas of the turtle Chrysemys dorbigni treated with streptozotocin.

The diabetogenic action of streptozotocin (SZ) was investigated in the turtle Chrysemys dorbigni after a 1- or 14-day fast. SZ (130 or 250 mg/kg) was injected intravenously, and blood glucose and plasma insulin were measured. Pancreatic endocrine cells were stained immunohistochemically by the immunoperoxidase avidin-biotin-peroxidase complex method. Only 14% of the SZ-treated turtles showed hyperglycemia. Prolonged fasting did not increase the percentage of hyperglycemic animals. In control turtles, insulin (beta)-, glucagon (alpha)- and somatostatin (delta)-immunoreactive cells were detected in increasing order of frequency. The qualitative changes seen in cells from the hyperglycemic SZ-treated turtles were more evident in beta and delta cells.

Animals