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Biomedical subjects

E A Nunez

Publications and source records attributed to E A Nunez.

At least 19 recordsLinked to original sources
PubMed1979

Relations between fatty acids and oestrogen binding properties of pure rat alpha 1-foetoprotein.

A delipidation procedure based on treatment with charcoal at pH 3 has been applied to highly purified rat alpha 1-foetoprotein preparations. The oestrogen binding properties of the delipidated proteins have been studied with an equilibrium dialysis technique, and compared with the properties of the untreated foetal protein, as well as those of preparations reconstituted from the defatted alpha 1-foetoprotein and the removed lipids. An important increase has been evidenced for the binding levels of oestrone, oestradiol-17 beta and diethylstilboestrol by the delipidated alpha 1-foetoprotein. A reversal of this effect has been obtained by incubating the delipidated protein either with the lipids extracted from the purified alpha 1-foetoprotein or with a potent competitor of the rat alpha 1-foetoprotein-oestrogen interaction, designated as 'L', previously demonstrated and isolated from whole rat sera, and tentatively characterized as a mixture of fatty acids. Scatchard analysis of the oestrone and oestradiol-17 beta binding parameters show that the enhanced fixation of the hormones after defatting is primarily due to a two-fold increase of the apparent number of binding sites/mol alpha 1-foetoprotein. The results are interpreted in terms of the probable, at least partial, identity between the lipids closely associated with the pure alpha 1-foetoprotein and the fatty acid mixture 'L' isolated from whole sera. The possible biological role of complex interplay between oestrophilic alpha 1-foetoproteins, phenolsteroids and fatty acids in the control of oestrogen levels during development is discussed briefly.

Animals↗
PubMed1979

Localization of a highly specific neuronal protein, serotonin binding protein, in thyroid parafollicular cells.

A highly specific serotonin binding protein (SBP) has been found in serotonergic neurons in both brain and gut. This protein has an extremely high affinity for serotonin and may be a storage protein. Serotonin is found in many endocrine cells, including parafollicular cells of the sheep thyroid, as well as in neurons. SBP is also present in sheep thyroid. The present study was done to localize the protein in the gland. Thyroid glands were divided into five segments. Concentrations of serotonin and SBP, as well as parafollicular cell volume were measured in each. Serotonin was assayed by enzymatic conversion to melatonin using tritiated S-adenosylmethionine. SBP was assayed by molecular sieve chromatography on sephadex G-50. The relative volume of parafollicular cells was obtained by stereological analysis of electron micrographs. Experiments were also done to demonstrate these cells by histofluorescence and radioautography following incubation with tritiated 5-hydroxytryptophan. Good correlations were found between serotonin and SBP concentrations, and parafollicular cell volume. These peaked in the rostro-central portion of the gland and were minimal at the poles. We conclude that thyroid SBP is probably localized in parafollicular cells.

Animals↗
PubMed1979

Ligand properties of diethylstilbestrol: studies with purified native and fatty acid-free rat alpha 1-fetoprotein and albumin.

We report the equilibrium binding parameters for the interactions of the estrogen analogue diethylstilbestrol (DES) with highly purified rat alpha 1-fetoprotein (AFP) and serum albumin preparations. At 25 degrees C and pH 7.4, an association constant (Ka) of about 1.5 X 10(6)M-1 and 2 sites/mole are measured with the DES-AFP system, whereas for the DES-albumin interaction, we find a Ka of approximately 2 X 10(5)M-1 and about 11 sites/mole of protein. The removal of fatty acids from pure AFP causes a reversible 3 fold increase of the number of DES binding sites; the same delipidation procedure applied to albumin slightly diminishes its DES binding parameters. We also demonstrate the capability of DES to displace competitively estradiol-17 beta (E2) from its high affinity sites on the estrophilic rat AFP. Finally, the binding behaviour of the two serum proteins towards the synthetic estrogen is compared to their interaction with the natural hormones. The physiological and pharmacological relevance of these data is discussed.

Animals↗
PubMed1979

An electron microscopic investigation of human familial bone dysplasia. Inhibition of osteocytic osteolysis and induction of osteocytic formation of elastic fibers following calcitonin treatment.

Familial bone dysplasia with hyperphosphatasemia is characterized by excessive bone resorption early in life with resulting severe skeletal deformity. The disease can be ameliorated by treatment with human calcitonin. We have the studied the ultrastructure of bone from diseased patients before treatment and at intervals during 1 year of treatment with calcitonin. Pretreatment osteoblasts, osteoclasts, and osteocytes exhibited mitochondria which contained vast amounts of dense microcrystal deposits. Osteocytes were also distinguished by minimal organellar development. Osteoclasts were rare. Calcitonin treatment included a progressive development of a more normal bone structure. Intramitochondrial crystal deposits were absent in mitochondria of osteocytes and osteoclasts but were still present in mitochondria of osteoblasts. Surprisingly, the developing bony matrix during calcitonin treatment exhibited large numbers of elastic fibers. These appeared to develop normally in alignment with the surface membrane of osteocytes. Calcitonin treatment caused a proliferation of osteocyte organellar development. It is concluded that familial bone dysplasia is primarily a disease of osteocytes and that osteocytic activity is influenced by calcitonin.

Bone Diseases, Developmental↗
PubMed1978

Formation of apical pseudopods by canine thyroid follicular cells: induction by thyrotropin and 5-hydroxytryptamine; antagonism by reserpine.

The role of biogenic amines in the activation of thyroid follicular cells by thyrotropin (TSH) was studied. 5-hydroxytryptamine (5-HT) was chosen as the amine to study and apical pseudopod formation, assessed by scanning electron microscopy, was used as the index of follicular cell activation. All experiments were done on dogs. TSH and 5-HT were both potent inducers of pseudopod formation. The action of TSH but not that of 5-HT was antagonized by the amine depleting drug reserpine. Reserpine depleted the thyroid of 5-HT in newborn, adolescent, and adult dogs. It is concluded that one or more biogenic amines, such as 5-HT, are probably involved in follicular cell activation by TSH.

Animals↗
PubMed1978

Calcium-induced release of 5-hydroxytryptamine from thyroid lobes in vitro and accompanying ultrastructural changes in parafollicular and follicular cells.

Biogenic amines, including serotonin (5-HT), have been shown to activate follicular cells of the thyroid. 5-HT is stored in bat's parafollicular cells. Previous radioautographical evidence indicates that this 5-HT is present in calcitonin granules. The present study was done to determine if Ca++, the natural stimulus to calcitonin release, would also release parafollicular cell 5-HT and, if so, whether this release would be accompanied by activation of follicular cells. Parafollicular cells were filled with labeled 5-HT by incubation of thyroid lobes of bats or mice with 5-[3H]hydroxytryptophan, the precursor of [3H]5-HT. Thyroid lobes were incubated in vitro in Ca++-free medium containing a chelating agent and were then challenged with Ca++ (0-30 mM). Release of [3H]5-HT was defected beginning at 5 mM Ca++. [3H]5-HT release was roughly proportional to the Ca++ concentration. Calcium challenge also affected thyroid ultrastructure in bats. After 10-min exposure to 30 mM Ca++, there were an increased number and centripetal movement of follicular cell lysosomes, development of apical pseudopods, and formation of colloid droplets. Many lysosomes also developed a crystalline-like matrix. An unique membrane-enclosed, rod-shaped organelle appeared in a small number of follicular cells. Parafollicular cells exposed to high concentrations of Ca++ were often degranulated, although many appeared unchanged. These changes confirm that 5-HT is released from parafollicular cells by elevating the external Ca++ concentration, supporting the hypothesis that 5-HT and calcitonin share storage granules. 5-HT release may mediate the activation of follicular cells by Ca++.

Animals↗
PubMed1977

Rat iso-alpha1-fetoproteins. Purification and interaction with estradiol-17beta.

The combination of polyacrylamide gel electrophoresis and Concanavalin-A-Sepharose affinity chromatography has permitted the isolation on a preparative scale, of four molecular forms of rat alpha 1-fetoprotein: a "slow" and a "fast" fraction, each separable into Concanavalin-A-adsorbed ("high carbohydrate", i.e. rich in accessible alphaD-Mannosyl and alphaD-Glu-cosyl residues) and a Concanavalin-A-non adsorbed ("low carbohydrate") fractions. These four iso-alpha 1-fetoproteins (iso-AFP) bind estradiol-17beta. However, they disclose differences in both their association constants and number of binding sites for this hormone. Very high affinity sites (10(9) are mainly located on the "slow-low carbohydrate" form. Low affinity, high capacity sites are preferentially located on the "high carbohydrate" form. These results confirm the molecular and functional heterogeneity of rat AFT and suggest that the carbohydrate moiety of the protein may have a role in estrogen-AFP interactions.

Animals↗
PubMed1977

Rat and human embryo and post-natal sera contain a potent endogenous competitor of estrogen-rat alpha-fetoprotein interactions.

A highly active inhibitor of the binding of estrone and estradiol-17beta to rat alpha-fetoprotein is demonstrated for the first time in embryo, immature and adult rat sera as well as in fetal and adult human sera. The competitive character and the narrow specificity of this inhibition effect is shown. The major compound responsible for this activity is isolated by successive column Sephadex LH20 and thin layer chromatography: it is characterized as a nonpolar, nonphenolic, dialysable and thermostable substance, unreactive towards anti-estrone and anti-estradiol-17beta antibodies. The possible biological role of an endogenous non-estrogen ligand of rodent fetoproteins is discussed.

Adrenalectomy↗
PubMed1977

Calcitonin treatment in hereditary bone dysplasia with hyperphosphatasemia: a radiographic and histologic study of bone.

Two children with bone dysplasia with hyperphosphatasemia (juvenile Paget's disease) were treated with synthetic human calcitonin. The progress of bone disease was monitored radiographically and histologically. Pretreatment radiographs showed markedly abnormal bone, characterized by lack of discrete cortex and absence of normal modeling. During treatment a discrete cortex was formed composed of compact bone, and more normal modeling occurred. This coincided with a histologic change from woven bone before treatment to a more lamellar type during treatment.

Alkaline Phosphatase↗
PubMed1976

Secretion by parafollicular cells beginning at birth: ultrastructural evidence from developing canine thyroid.

The ultrastructure of developing canine parafollicular cells has been examined. The hypothesis that secretion is relatively inactive prior to birth but very active at and following birth was tested. Parafollicular cells develop and accumulate characteristic secretory granules prior to birth. However, there is little or no evidence of exocytosis at this time. At birth and during the neonatal period, but not in adult thyroids, signs of exocytosis of granular contents by parafollicular cells are abundant. Just prior to the expected date of birth and before evidence of exocytosis appears, parafollicular cells accumulate intracisternal granules within rough endoplasmic reticulum. These observations are consistent with the view that parafollicular cells first become actively secretory around the time of birth and are more active at this time than in early fetal or later adult life.

Age Factors↗
PubMed1976

Purification and interaction with estradiol-17beta.

The combination of polyacrylamide gel electrophoresis and Concanavalin-A-Sepharose affinity chromatography has permitted the isolation on a preparative scale, of four molecular forms of rat alpha1-fetoprotein: a "slow" and a "fast" fraction, each separable into Concanavalin-A-adorbed ("high carbohydrate", i.e. rich in accessible alphaD-Mannosyl and alphaD-Glucosyl residues) and a Concanavalin-A-non adsorbed ("low carbohydrate") fractions. These four iso-alpha-fetoproteins (iso-AFP) bind estradiol-17beta. However, they disclose differences in both their association constants and number of binding sites for this hormone. Very high affinity sites (10(9)) are mainly located on the "slow-low carbohydrate" form. Low affinity, high capacity sites are preferentially located on the "high carbohydrate" form. These results confirm the molecular and functional heterogeneity of rat AFP and suggest that the carbohydrate moiety of the protein may have a role in estrogen-AFP interactions.

Animals↗
PubMed1975

Lipid droplet accumulation in cardiac muscle cells of the bat: potential auto-toxicity of the cardiac sympathetic innervation.

The previously described ability of reserpine and parachlorophenylalanine to induce the accumulation of lipid droplets in ventricular cardiac muscle cells of the bat was investigated. Lipid droplet accumulation was assessed qualitatively by light microscopy and quantitatively by morphometric analysis of electron micrographs. An hypothesis that the action of the drugs was an indirect one, mediated by the cardiac adrenergic innervation, was framed and tested. Lipid droplet accumulation occurred during a time of intense sympathetic activity, that of arousal from hibernation. The ability of the two drugs to produce the effect was antagonized by prior sympathetectomy with 6-hydroxy-dopamine. The effect was mimicked by administration of exogenous norepinephrine together with inhibitors of its catabolic enzymes, monoamine oxidase and catechol-o-methyl transferase. These observations are all consistent with the initial hypothesis and raise the possibility that endogenous norepinephrine in the cardiac sympathetic innervation might be, at least potentially, auto-toxic.

Animals↗
PubMed1975

Paracrystalloids from mitochondria in thyroid follicular cells. Normal occurrence and experimental induction.

The ultrastructure of the follicular cells of the thyroid gland of bats caught in early June, when the euthyroid state is being restored following the phase of great thyroid activity that occurs at arousal, was studied. Several ultrastructural features were found which had not been observed in animals caught at other periods of the annual life cycle. The most striking ultrastructural feature was the presence in many cells of prominent, large membrane-enclosed paracrystalloid bodies. The paracrystalloids exceeded 6.0 mum in length and were approximately 1.0 mum. in width. Examination of transition forms showed that paracrystalloids were derived from mitochondria and were formed by the reorganization of the cristae into parallel closely apposed membranes. The membranes run longitudinally in loops from one end of the mitochondrion to the other. Similar mitochondrial alterations were observed in rats by duplicating the thyroid conditions of the June bat. Rats were given thiouracil for 10 weeks followed by L-triiodothyronine injection. These animals all showed similar mitochondrial-paracrystalline alterations which were not seen in hypophysectomized animals or animals given L-triiodothyronine alone. Mitochondria from thyroids of rats given thiouracil alone showed an increase in size and number and changes in shape and internal structure. The results of the bat and rat studies suggest that rapid shutdown of active follicular cells appears to promote thyroid paracrystalloid formation from mitochondria.

Animals↗