PubMed Health⌕ Search

Biomedical subjects

F J Alvarez

Publications and source records attributed to F J Alvarez.

At least 55 records · Page 3Linked to original sources

Calbindin D28k expression in immunohistochemically identified Renshaw cells.

Double immunofluorescence was utilized to determine whether Renshaw cells contain calbindin D28k immunoreactivity. Renshaw cells were identified by their characteristic expression patterns of gephyrin immunoreactivity in sections of rat and cat lumbar spinal cord. In the rat, all neurons classified as Renshaw cells (n = 487) also contained calbindin D28k-immunoreactivity, and all calbindin D28k-immunoreactive cells located in the ventral-most region of lamina VII expressed the characteristic gephyrin labeling and morphology of Renshaw cells. In the cat, fewer than half of the Renshaw cells (47%; n = 128) were double-labeled. In both species, occasional calbindin D28k-immunoreactive Renshaw cells were identified within motor nuclei in lamina IX. The distinctive immunolabeling of Renshaw cells allowed us to estimate that there are about 250 Renshaw cells in each ventral horn of the fourth lumbar segment of rat spinal cord, and about 750 cells per ventral horn in the L6 segment of the cat. We conclude that the functional properties of Renshaw cells, including their ability to fire action potentials at high rates, likely require specific homeostatic mechanisms including strong intracellular calcium buffering, the precise mechanisms of which may vary between species.

Animals↗

Distribution of 5-hydroxytryptamine-immunoreactive boutons on alpha-motoneurons in the lumbar spinal cord of adult cats.

Recent studies have shown that at least some of the functional effects of serotonin (5-HT) on motoneuron excitability are direct and are mediated via postsynaptic 5-HT receptors on motoneurons. To determine the spatial distribution of direct inputs from the serotonin system on the proximal and distal dendrites of individual motoneurons, we examined identified motoneurons in vivo with a combination of immunohistochemical localization of 5-HT-immunoreactive boutons and intracellular staining with horseradish peroxidase. Seventeen intracellularly stained motoneurons from 12 adult cats were analyzed with light microscopy. Quantitative analysis of 5-HT boutons apposed to dendrites of five representative motoneurons that were entirely reconstructed in three dimensions (each from the lumbosacral spinal cord of a different animal) revealed a total of 7,848 contacts (1,570+/-487 contacts/postsynaptic neuron; mean +/- SD) over the dendrites of these cells. Analysis of contacts on the soma of two of these cells, and on the somas of an additional 12 intracellularly stained motoneurons, revealed a wide range of somatic contacts (11-211 contacts/cell) on motoneuron cell bodies, with an average of 52 contacts/cell. These results indicate that the vast majority of 5-HT-immunoreactive boutons are apposed to dendritic branches rather than to the somatic surface of motoneurons. The spatial distribution of contacts essentially matched the distribution of surface membrane area of the postsynaptic neuron, resulting in a relatively uniform density of contacts (<1/100 microm2) on proximal and distal dendrites. Consequently, the frequency of contacts was higher on the proximal dendritic compartments where available membrane area is greater. There was no preferential distribution of contacts to particular dendrites. Light/electron microscopic correlations were performed on 21 boutons that contacted dendrites (n = 7) of three motoneurons from different animals. At the electron microscope level, most appositions (18/21; 85.7%) selected by our light microscopic criteria were confirmed as direct contacts when the 5-HT boutons were examined through serial sections. Synaptic junctions, generally small and symmetric, were positively identified in only a subset of these cases (n = 6; 28.6%), in part due to the obscuring effects of the peroxidase histochemical precipitate present in both pre- and postsynaptic profiles. A few 5-HT boutons (3/21; 14.3%) selected as contacts by our light microscopic criteria were in fact separated from the adjacent labeled dendrites; in two of these three cases, the separation was due to intrusion of very thin glial lamellae (<0.3 microm in cross section). These results indicate that the bulbospinal serotonergic system(s) provide a significant, direct synaptic input to spinal motoneurons that innervate hindlimb muscles. The nature of the modulatory actions exerted by such widespread synaptic inputs will affect all regions of the somatodendritic membrane and will ultimately depend on the nature of the 5-HT receptors present over different parts of the postsynaptic neuron's dendritic tree.

Animals↗

Effect of sciatic nerve transection or TTX application on enzyme activity in rat spinal cord.

To clarify the differential effects on spinal circuitry caused by physical vs functional disconnection from the periphery, we compared changes produced by 3-, 7- or 14-day unilateral sciatic axotomy or tetrodotoxin (TTX) nerve blockade on the abundance or activity of NADPH diaphorase (NDP), cytochrome oxidase (CO) and acid phosphatase (AP) in the spinal cord. Following axotomy, AP and NDP were decreased in the dorsal horn and increased in large cells in the dorsolateral motor nuclei while CO was decreased in ventral horn neuropil. TTX induced a decrease of CO in the ventral horn and NDP in the dorsal horn. This suggests that physical vs functional disconnection causes modulation of distinct intracellular pathways in sensory afferents, dorsal horn neurons and motoneurons.

Acid Phosphatase↗

Comparison of rapid bolus instillation with simplified slow administration of surfactant in lung lavaged rats.

The aim of this study was to compare the effects of modified porcine surfactant (Curosurf) given either by a simplified slow delivery technique or by the standard bolus method, on pulmonary gas exchange, lung mechanics, and surfactant distribution in rats with respiratory failure produced by lung lavage. Twelve rats with respiratory failure induced by lung lavage received 200 mg x kg(-1) body weight (b.w.) of tagged porcine surfactant, either by the standard bolus delivery technique or by a simplified 1-min intratracheal infusion method, not requiring interruption of mechanical ventilation. Cardiovascular parameters, arterial blood gases, and pulmonary mechanics were measured repeatedly. Surfactant distribution was also measured by dye-tagged microbead spheres. After surfactant administration, there were no overall major differences between groups in mean heart rate, blood pressure, arterial blood gases, dynamic lung compliance, respiratory system resistance, and pulmonary distribution of exogenous surfactant. However, after 180 min pulmonary gas exchange was better and compliance higher in the bolus than the 1-min infusion group. A transient decrease in blood pressure and heart rate was observed in the bolus group; this side effect was not seen in animals treated with the simplified 1-min infusion method. We conclude that in rats subjected to lung lavage, the infusion of porcine surfactant by a simplified 1-min procedure produced similar short-term effects compared to the same dose of surfactant given by the bolus method. We speculate that tracheal bolus dosing is highly effective and might be the preferable delivery method for porcine surfactant. Dosing by the simplified method described appears less effective, but since no significant differences were observed, and since it produced less acute adverse effects, it could be used when clinical circumstances preclude rapid delivery.

Animals↗

Diversity of structure and function at mammalian central synapses.

Our appreciation of the relationship between synaptic structure and function, and in particular our understanding of quantal synaptic transmission, is derived from classical studies on the neuromuscular junction. However, physiological studies of quantal transmission at mammalian CNS synapses have produced a variety of results, and thus no consensus of opinion has emerged. This variability could be due, in part, to experimental and analytical limitations or to differences in the structural and functional features of central synapses, or both. Some of the experimental limitations have recently been overcome by the use of novel preparations that permit direct measurement of quantal synaptic events in the CNS. Although these studies reveal similarities between the synaptic mechanisms of the neuromuscular junction and CNS synapses, important differences and specializations are also evident. The purpose of this review is to highlight the structural and functional diversity of synapses in the mammalian CNS, and to discuss the potential relevance of structural features to synaptic function.

Animals↗

In vivo behaviour of rat band 3 cross-linked carrier erythrocytes.

Rat band 3 cross-linked carrier erythrocytes have been prepared. Iodinated carbonic anhydrase has been encapsulated into rat erythrocytes. Then, carrier erythrocytes were labeled with 51chromium. Eventually, these doubly labeled rat RBCs were treated with a band 3 cross-linking reagent, namely bis(sulfosuccinimidyl)suberate (BS3). 51Chromium labeling and 125I CA showed to have cytosolic localization in cross-linked carrier erythrocytes. Estimation of the band 3 cross-linking induced by BS3 on rat carrier erythrocytes has been done rendering values around 25% of band 3 monomer reduction. BS3-cross-linked carrier erythrocytes when injected into rats are mainly targeted to liver as shown by chromium labeling localization. Also, encapsulated CA radioactivity carried by cross-linked carrier rat erythrocytes when injected into rats is localized predominantly in liver as shown by in vivo experiments. Accordingly, cross-linked carrier erythrocytes are highly recognized by peritoneal macrophages as detected by in vitro analyses of macrophage recognition. Thus, our data revealed a targeting of carrier rat erythrocytes induced by cross-linking of band 3 protein by BS3. These results support claims in favor of this animal model as a feasible system to analyze cross-linked carrier erythrocytes survival and targeting as well as the in vivo efficacy of targeting of loaded compounds to liver.

Animals↗

Hypotonically loaded rat erythrocytes deliver encapsulated substances into peritoneal macrophages.

Previous work has shown increased uptake of hypotonically loaded rat RBCs by the spleen and liver "in vivo," suggesting that the cells of MPS are involved in their elimination from the circulation. In order to elucidate the mechanism of such elimination, we have undertaken studies on the interaction of such loaded RBCs, in comparison with native RBCs, with peritoneal macrophages. Erythrophagocytosis assays were performed in well plates to which thioglycollate-induced peritoneal macrophages had adhered. Native or loaded 51Cr-RBCs were added under different opsonization conditions to monolayer adherent macrophages, and then the amount of RBCs that were recognized was determined, with separation into adhesion and phagocytosis fractions. Native RBCs are slightly recognized by peritoneal macrophages, about one RBC per macrophage (Mphi). Osmotic treatment of rat RBCs used for encapsulation (independently of the encapsulated substance, 125I-CA or FITC-dextran) produces some modification in the erythrocyte membrane that induces higher recognition of these cells, about three loaded RBCs per macrophage. Consequently, both fluorescent (FITC-Dx) and radioactive (125I-CA) substances previously encapsulated in RBCs were transferred to M(phi)s. The fluorescence microscopic observations confirmed these results. Moreover, in the case of carrier 51Cr-cells loaded with 125I-CA, the amount of 125I-radioactivity delivered into M(phi)s was relatively higher than that of 51Cr. The highest ratio, 125I-CA (encapsulated substance)/51Cr-RBCs (carrier cells), present in M(phi)s means there was a stronger interaction with macrophages of RBCs that carry a higher amount of encapsulated CA, as a function of the heterogeneity of the loaded rat RBCs population previously reported. Finally, the adhesion and phagocytosis of loaded RBCs seem not to involve complement receptors or Fc receptors on the macrophages.

Animals↗

Differential induction of macrophage recognition of carrier erythrocytes by treatment with band 3 cross-linkers.

Mouse native and hypotonically loaded erythrocytes were treated with two cross-linking reagents: bis(sulphosuccinimidyl)suberate (BS3)- and 3,3'-dithiobis-(sulphosuccinimidyl propionate) (DTSP), excluding clustering agents. Microscopic analyses revealed that band 3 cross-linked native and hypotonically loaded erythrocytes are more strongly recognized by peritoneal macrophages than native and loaded erythrocytes as a result of the cross-linking of band 3 protein in accordance with studies in vivo. Macrophage-recognition analyses of 51Cr-labelled erythrocytes also demonstrated increased recognition of cross-linked and cross-linked loaded erythrocytes. This shows that the only action of these two band 3 cross-linkers on mouse erythrocytes promotes recognition by macrophages without requiring the use of clustering agents. The extent of recognition of BS3 cross-linked and cross-linked loaded erythrocytes by macrophages is dependent on the presence or absence of homologous serum or immunoglobulins. In contrast, the presence of serum factors or IgG in the incubation medium did not seem to influence the recognition of DTSP-modified erythrocytes by macrophages. These results seem to indicate a different mechanism of recognition for the erythrocytes modified with either one or the other band 3 cross-linker. In summary, the unique use of both band 3 cross-linkers procedures can be used to target carrier erythrocytes conveying active compounds to macrophages, with possible therapeutical applications. Different mechanisms of induction of macrophage recognition by these band 3 cross-linkers could reveal differential actions on erythrocytes or the involvement of different factors in the recognition process.

Animals↗

Cross-linking treatment of loaded erythrocytes increases delivery of encapsulated substance to macrophages.

Previous investigation has shown that osmotically loaded erythrocytes can act as drug carriers in systemic circulation, whereas chemically modified erythrocytes can be targeted to organs of the mononuclear phagocytic system because of changes introduced in the membrane that are recognized by macrophage cells. In this study we have examined the delivery of 125I-labelled carbonic anhydrase (125I-CA) carried by mouse erythrocytes, either loaded, or loaded and cross-linked with bis(sulphosuccinimidyl)suberate (BS3) and 3,3'-dithiobis-(sulphosuccinimidyl propionate), into homologous peritoneal macrophages maintained in culture. The hypotonically loaded mouse erythrocytes show a slight recognition by macrophages, similar to native erythrocytes. CA loaded into erythrocytes is thus delivered to a limited extent into macrophages. Neither the number of recognized loaded 51Cr-labelled erythrocytes nor the amount of delivered 125I-CA is affected by the presence of serum components or IgG. In contrast, cross-linking these loaded erythrocytes results in a greater phagocytosis by macrophages as assessed by microscopic observations, producing a markedly increased amount of targeted enzyme. The amount of CA delivered into macrophages, after BS3 cross-linker treatment of erythrocytes, is dependent on the presence of serum components in the incubation medium. Thus these cross-linking treatments improve the capacity of loaded mouse erythrocytes to deliver significant amounts of targeted enzyme to macrophage cells, increasing the therapeutic potential of carrier erythrocytes.

Animals↗

Downregulation of metabotropic glutamate receptor 1a in motoneurons after axotomy.

Axotomized motoneurons display drastic modifications in synaptic structure and function related to their disconnection from the periphery and establishment of a regenerative metabolic functional mode. The molecular basis of these modifications is not fully understood. Here we describe changes in metabotropic glutamate receptor 1a (mGluR1a)-immunoreactivity 3, 7 or 14 days after unilateral aciatic transection. mGluR1a-immunoreactivity was distributed throughout the somatic cytoplasm and somatodendritic membrane of uninjured motoneurons and was significantly reduced in axotomized motoneurons. This reduction was observed at 3 days and grew progressively over 2 weeks. These findings suggest that downregulation of mGluR1a could contribute to reduced excitatory neurotransmission in axotomized motoneurons.

Animals↗

Cell-type specific organization of glycine receptor clusters in the mammalian spinal cord.

Glycinergic synapses play a major role in shaping the activity of spinal cord neurons. The spatial organization of postsynaptic receptors is likely to determine many functional parameters at these synapses and is probably related to the integrative capabilities of different neurons. In the present study, we have investigated the organization of gephyrin expression along the dendritic membranes of alpha- and gamma-motoneurons, Ia inhibitory interneurons, and Renshaw cells. Gephyrin is a protein responsible for the postsynaptic clustering of glycine receptors, and the features of gephyrin and glycine receptor alpha(1)-subunit immunofluorescent clusters displayed similar characteristics on ventral horn spinal neurons. However, the density of clusters and their topographical organization and architecture varied widely in different neurons and in different dendritic regions. For motoneurons and Ia inhibitory interneurons, cluster size and complexity increased with distance from the soma, perhaps as a mechanism to enhance the influence of distal synapses. Renshaw cells were special in that they displayed an abundant complement of large and morphologically complex clusters concentrated in their somas and proximal dendrites. Serial electron microscopy confirmed that the various immunoreactivity patterns observed with immunofluorescence accurately parallel the variable organization of pre- and postsynaptic active zones of glycinergic synapses. Finally, synaptic boutons from single-labeled axons of glycinergic neurons (Ia inhibitory interneurons) were also associated with postsynaptic receptor clusters of variable shapes and configurations. Our results indicate that mechanisms regulating receptor clustering do so primarily in the context of the postsynaptic neuron identity and localization in the dendritic arbor.

Animals↗

The use of medication by the Spanish population.

The aim of the study was to assess patterns of the use of medicines by the general population older than 0 years. The study was based on the information contained in the computerized database from the 1993 Spanish Household Health Survey. A representative sample of the population older than 0 was identified, and a survey of 26,334 persons was carried out. Of the population 45.3% had taken some medicine in the last 2 weeks prior to the carrying out of the survey. The proportion was greater for women (50.6%) than for men (39.6%). With increasing age, the frequency and amount of medication use increased. A little over 20% of the medicines used were not prescribed by a doctor (self-medication). Data show the frequency of the use of medicines by the Spanish population.

Journal Article↗

Thyroid hormones in the pathogenesis of lung hypoplasia and immaturity induced in fetal rats by prenatal exposure to nitrofen.

BACKGROUND/PURPOSE: Nitrofen is believed to act on prenatally exposed fetuses by changing maternal or fetal thyroid hormone physiology. The aim of this study was to determine whether the amounts of circulating and lung tissue T3 and T4 are decreased in rat fetuses with nitrofen-induced pulmonary hypoplasia and diaphragmatic hernia. METHODS: Timed-pregnant rats were given 100 mg of nitrofen in oil on gestational day 9.5, and their fetuses were recovered on the 21st day. Lung weight to body weight ratio was determined. Hormonal studies consisted in measurement of plasma T3, T4, and TSH, and of T3, T4, and DNA in lung tissue. Suitable groups of control fetuses prenatally exposed to oil were used for comparison. RESULTS: The lungs of nitrofen-treated fetuses were hypoplastic and those who had congenital diaphagmatic hernia were even more so. Nitrofen treatment led to decreased plasma T3 and T4 levels without TSH changes. T3 and T4 in lung tissue were apparently decreased in treated fetuses when expressed by weight, but these differences disappeared when expressed by DNA (cell content). CONCLUSIONS: Lung hypoplasia and immaturity induced by nitrofen treatment are not related to decreased levels of thyroid hormones in tissue near term. This should be kept in mind when proposing hormonal treatment for prenatal induction of lung maturation.

Abnormalities, Drug-Induced↗

Maternal exposure to delta 9-tetrahydrocannabinol (delta 9-THC) alters indolamine levels and turnover in adult male and female rat brain regions.

Perinatal exposure to delta 9-THC has been shown to produce effects on brain development. In this study we evaluated the changes induced by maternal exposure to delta 9-THC (5 mg/kg per day) from gestational day 5 to postnatal day 24 in eight discrete brain areas on the central serotoninergic system in both adult male and female rats. These result show that maternal exposure to delta 9-THC from gestational day 5 to postnatal day 24 affects development of the various central indoleaminergic system of the offsprings brain. Perinatal exposure to delta 9-THC decreased the levels of 5-HT in hypothalamus and rostral neostriatum in exposed males, and also decreased the levels of 5-HT in ventral hippocampus, septum, and midbrain raphe nuclei in both exposed males and females. Perinatal exposure to delta 9-THC increased the levels of 5-HIAA in dorsal hippocampus, hypothalamus, septum, midbrain raphe nuclei, and rostral neostriatum in exposed males and females. We have also found differences between nonexposed males and females in several brain regions. Our results confirm a regional and sexual specificity in endogenous levels of indoleamine after perinatal delta 9-THC treatment, being the midbrain raphe nuclei the most affected area.

Animals↗

Effect of maternal delta 9-tetrahydrocannabinol on developing serotonergic system.

In this study we investigated the effects of maternal delta 9-tetrahydrocannabinol on the developing serotonergic system. A daily dose of delta 9-tetrahydrocannabinol (5 mg/kg body weight) was administered p.o. to pregnant rats from gestational day 5 to postnatal day 1. Levels of indolamines were measured in four brain areas of the offspring on the day before or after birth. Levels of indolamines depended on the cerebral area, sex and pre- or postnatal age. Maternal exposure to delta 9-tetrahydrocannabinol decreased diencephalic levels of 5-hydroxytryptamine (5-HT), males being more susceptible than females. These perinatal changes could be responsible for the long-term neurophysiological alterations produced by cannabinoids.

Animals↗

[Contraceptive use and smoking among Spanish women].

OBJECTIVE: The aim of this study is to analyze the oral contraceptive use and smoking habits among Spanish women. DESIGN: Descriptive observation study of a crossover design. SETTING: General population, using to this purpose the 1993 National Household Health Survey. PATIENTS AND OTHER PARTICIPANTS: 5229 Spanish women aged 16-42 years. MEASUREMENTS AND MAIN RESULTS: The use of oral contraceptives, as well as patterns of smoking was assessed. 4.6% of the women of 16-42 years of age had used oral contraceptives in the last two weeks, from which 51.1% smoked in a daily basis. Furthermore, mean number of cigarettes/day and heavy smokers was higher among oral contraceptive users than in non users. CONCLUSIONS: Smoking habit and oral contraceptive use is frequent among Spanish women (this population a higher risk of certain side effects). Health professionals should be aware of smoking the women taking oral contraceptives.

Adolescent↗

Fluorescence analysis of carrier rat and human erythrocytes loaded with FITC-dextran.

Rat and human erythrocytes are inherently different with respect to slow dialysis encapsulation used in preparing carrier erythrocytes. The incorporation process, commonly measured with radioactive tracers, is always larger in human erythrocytes, mainly because the rat carrier cells are more fragile. When FITC-Dextran (Dx) is used in the encapsulation process, and loaded rat and human RBCs are studied by fluorescence intensity, some additional events are evident. Not all cells of each population appear with a fluorescence signal, and not all show similar fluorescence intensity. Human RBCs show a higher percentage of marked cells and a higher fluorescence intensity than rat RBCs. Two populations, of high and low fluorescence, appear in FITC-Dx loaded rat erythrocytes. The human loaded RBCs show a similar peak distribution together with another peak in the middle scale of fluorescence. Therefore, a heterogeneity in the cell population as a result of the encapsulation process is manifested for both species. The fractionation of RBCs, loaded with either FITC-Dx or 125I-CA, by centrifugation on Ficoll-Paque reveals that the low density cells have much more substance incorporation than the counterpart cell subpopulation in the pellet. Therefore, the cell modifications produced by the encapsulation process are independent of the substance being incorporated. On the other hand, FITC-Dx, but not 125I-CA, shows a certain degree of association to RBCs membranes, especially in humans.

Animals↗