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Biomedical subjects

M Reyes

Publications and source records attributed to M Reyes.

At least 19 recordsLinked to original sources

Immune responses associated with chronic fatigue syndrome: a case-control study.

An exploratory case-control study was conducted to assess whether the many reported differences in the immune function of chronic fatigue syndrome (CFS) patients are detectable in rigorously defined cases of CFS. Although many studies have reported differences between cases and controls in various measures of immune function, none of these differences were found in all studies. In this study, no differences were found in white blood cell numbers; immune complex, complement, or serum immunoglobulin levels; delayed type hypersensitivity and allergic responses; NK cell function; and proliferative responses to mitogens and antigens. Marginal differences were detected in cytokine responses and in cell surface markers in the total CFS population. However, when the patients were subgrouped by type of disease onset (gradual or sudden) or by how well they were feeling on the day of testing, more pronounced differences were seen.

Adolescent

Induction of hippocampal long-term depression requires release of Ca2+ from separate presynaptic and postsynaptic intracellular stores.

Studies have suggested that an increase in intracellular [Ca2+] is necessary for the induction of both long-term potentiation (LTP) and long-term depression (LTD) of synaptic transmission, and that release of Ca2+ from intracellular storage pools can be necessary to induce LTP. We investigated whether release of Ca2+ from intracellular stores also is required for the induction of LTD at Schaffer collateral-CA1 synapses in hippocampal slices. Both thapsigargin (1 microM) and cyclopiazonic acid (1 microM), compounds that deplete all intracellular Ca2+ pools by blocking LTP-dependent Ca2+ uptake into intracellular compartments, blocked the induction, but not maintenance, of LTD by low-frequency stimulation (LFS) (1 Hz/15 min) without affecting baseline synaptic transmission. Washout of the reversible inhibitor cyclopiazonic acid restored the ability to induce LTD. In contrast, thapsigargin did not block depotentiation of LTP by 1 Hz LFS, suggesting that LTP causes a reduction in the threshold [Ca2+] necessary for LTD. Selective depletion of the ryanodine receptor-gated Ca2+ pool by bath application of ryanodine (10 microM) also blocked the induction of LTD, indicating a requirement for Ca(2+)-induced Ca2+ release. Impalement of CA1 pyramidal neurons with microelectrodes containing thapsigargin (500 nM to 200 microM) prevented the induction of LTD at synapses on that neuron without blocking LTD in the rest of the slice. In contrast, similar filling of CA1 pyramidal neurons with ryanodine (2 microM to 5 mM) did not block the induction of LTD. From these data, we conclude that the induction of LTD requires release of Ca2+ both from a presynaptic ryanodine-sensitive pool and from postsynaptic (presumably IP3-gated) stores.

Animals

Entamoeba histolytica: proteinase secretion induced by collagen type I is dependent on cytoskeleton integrity.

Proteolytic activities of the protozoan parasite Entamoeba histolytica strain HM1:IMSS and the cytochalasin D-resistant mutant BG-3 were analyzed following stimulation with collagen type I and Ca2+, which induces electron-dense associated collagenase secretion. The mutant BG-3 had a protease activity of 73 kDa and secretion of total protease activity was not stimulated by collagen type I and Ca2+, which produced, in contrast, a 2-fold increase in protease secretion by the parental strain. This collagen-stimulated protease secretion was inhibited by cytochalasin D at a concentration of 1 microgram/ml. Cytochalasin D did not have any effect on the protease activity released by the mutant BG-3. These findings suggest that cytoskeleton integrity is necessary for collagen-induced protease secretion.

Animals

Respiratory infection and iatrogenic diarrhea in Honduras and El Salvador during the 1991-1992 season.

The etiology of acute respiratory tract infection and its association with diarrhea was analyzed in 135 hospitalized children less than three years of age with mainly respiratory symptoms in two pediatric hospitals in Honduras and El Salvador. Etiologic diagnoses were performed on nasopharyngeal samples by tissue culture and immunofluorescence, including a search for the presence of respiratory virus-specific immunoglobulin A antibodies. Fecal samples were subjected to electron microscopy and tissue culture. The majority (83%) of the children (65% boys and 35% girls) were less than 12 months old and 45% were less than four months old. In 63 (47%) patients, at least one viral infection of the respiratory tract was identified. Respiratory syncytial virus (RSV) or the respective IgA antibodies were found in 43 (32%) patients., influenza A and B viruses in 29 (21%) patients, adenovirus in four patients, and enterovirus in two patients. Twenty (15%) patients had two or more viral agents. An association between RSV cases and environmental temperature was established. In 124 fecal samples, we identified four cases with astrovirus, seven with nonpolio enterovirus, and eight with adenovirus, but only three cases with rotavirus. Diarrhea was present in 55 (45%) of the patients. There was a statistically significant association between diarrhea and cases with RSV. This was shown to be associated with antibiotic treatment.

Age Distribution

Functional properties of regenerated optic axons terminating in the primary olfactory cortex.

When the optic nerve of Rana pipiens is cut and deflected into the telencephalon, the regenerating fibers terminate selectively in the superficial neuropil of the primary olfactory cortex. These redirected fibers and their terminals on the dendrites of the cortical cells appear normal by LM and EM criteria. Electrical recording, done 2-16 months after surgery, shows visually evoked activity in the superficial neuropil (Layer I) of the olfactory cortex, and visually excited responses in the deep cortical cell layer (Layer II). In the normal frog, the electrical activity seen in the neuropil of the olfactory cortex consists of small transients about 2-3 x the noise level of the electrode contact. These occur spontaneously and are also excited by puffs of air to the nose. There is no such excitation by visual stimuli. Larger initially negative spikes cell above noise level are recorded in the cell layer next to the ependymal surface, and these are also spontaneous, or excited by puffs of air to the nose, but not by visual stimuli. In the operated frog, the small transients in the neuropil appear and are excited by the puffs of air and by visual stimuli. Similarly the responses in the cell layer are excited by both sorts of stimuli. But new types of electrical signals appear in the neuropil; they are driven only by visual stimuli presented to the affected eye. These are very large transients of the kind found in the tectal neuropil and have the two characteristic shapes which were classified as B and C types in the tectum. Such large transients are never seen in the neuropil of the olfactory cortex in normal frogs. The receptive fields of the small visually driven transients in the neuropil are not easy to make out because the signal levels are so close to the noise level that different units cannot be reliably distinguished from each other. But the receptive fields of the much larger B and C type unit responses are as easy to classify and plot as they are in tectum, even though on the average they are only about 2/3 as large as in tectum. The single-unit receptive fields belong to one or another of the several types of retinal ganglion cell classes distinguished in optic-nerve recordings. Four of the major classes normally project to the tectum and a fifth projects to the lateral geniculate complex. But all five are present in the ectopic projection to the olfactory cortex.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Identification of a new 6-bromo-5,9-eicosadienoic acid from the anemone Condylactis gigantea and the zoanthid Palythoa caribaeorum.

A new brominated fatty acid, 6-bromo-5,9-eicosadienoic acid [1] was identified in the phospholipids (mainly phosphatidylethanolamine) of the anemone Condylactis gigantea and the zoanthid Palythoa caribaeorum. The delta 5,9 fatty acids, 5,9-octadecadienoic acid, 5,9-eicosadienoic acid, 5,9-docosadienoic acid, and 5,9-tetracosadienoic acid, were also identified in both organisms. Structural elucidation was accomplished by spectroscopic and chemical means. Our results further corroborate that delta 5,9 phospholipid fatty acids are not unique to sponges, as recognized previously, but can be found in other marine invertebrates such as anemones and zoanthids. An improved procedure for the synthesis of picolinyl esters is also described.

Animals

Seroepidemiology of chronic fatigue syndrome: a case-control study.

We performed serological testing for a large number of infectious agents in 26 patients from Atlanta who had chronic fatigue syndrome (CFS) and in 50 controls matched by age, race, and sex. We did not find any agent associated with CFS. In addition, we did not find elevated levels of antibody to any of a wide range of agents examined. In particular, we did not find elevated titers of antibody to any herpesvirus, nor did we find evidence of enteroviral exposure in this group of patients.

Adolescent

Mutations that alter the transmembrane signalling pathway in an ATP binding cassette (ABC) transporter.

The maltose transport system of Escherichia coli is a well-characterized member of the ATP binding cassette transporter superfamily. Members of this family share sequence similarity surrounding two short sequences (the Walker A and B sequences) which constitute a nucleotide binding pocket. It is likely that the energy from binding and hydrolysis of ATP is used to accomplish the translocation of substrate from one location to another. Periplasmic binding protein-dependent transport systems, like the maltose transport system of E.coli, possess a water-soluble ligand binding protein that is essential for transport activity. In addition to delivering ligand to the membrane-bound components of the system on the external face of the membrane, the interaction of the binding protein with the membrane complex initiates a signal that is transmitted to the ATP binding subunit on the cytosolic side and stimulates its hydrolytic activity. Mutations that alter the membrane complex so that it transports independently of the periplasmic binding protein also result in constitutive activation of the ATPase. Genetic analysis indicates that, in general, two mutations are required for binding protein-independent transport and constitutive ATPase. The mutations alter residues that cluster to specific regions within the membrane spanning segments of the integral membrane components MalF and MalG. Individually, the mutations perturb the ability of MBP to interact productively with the membrane complex. Genetic alteration of this signalling pathway suggests that other agents might have similar effects. These could be potentially useful for modulating the activities of ABC transporters such as P-glycoprotein or CFTR, that are implicated in disease.

ATP-Binding Cassette Transporters

New 2-hydroxy fatty acids in the Caribbean urchin Tripneustes esculentus.

The novel alpha-hydroxy fatty acids 2-hydroxy-13-docosenoic acid [1a], 2-hydroxy-14-tricosenoic acid [2a], and 2-hydroxy-15-tetracosenoic acid [3a] were identified in the Caribbean urchin, Tripneustes esculentus. The double-bond positions of the novel alpha-hydroxy fatty acids were determined by derivatization with dimethyl disulfide and shown to correlate with the corresponding non-hydroxylated mono-unsaturated fatty acids, 13-docosenoic acid, 14-tricosenoic acid, and 15-tetracosenoic acid also present in T. esculentus. The total fatty acid composition of the urchins is also reported where cis-5-olefinic fatty acids such as 5,9-octadecadienoic acid and 5,11-eicosadienoic acid were found to predominate in the mixture. Cholesterol was the predominant sterol in T. esculentus.

Aldehydes

Characterization of the structural requirements for assembly and nucleotide binding of an ATP-binding cassette transporter. The maltose transport system of Escherichia coli.

The periplasmic maltose-binding protein-dependent, maltose transport system of Escherichia coli is a well studied member of the ATP-binding cassette family of transport ATPases. In addition to the water-soluble maltose-binding protein, the system comprises three membrane proteins, MalF, MalG, and MalK, which form a heterotetrameric complex (FGK2) in the cytoplasmic membrane. The purified complex exhibits transport-associated ATPase activity. To characterize the requirements for nucleotide binding and hydrolysis by the FGK2 complex, we used plasmids to express different combinations of the individual subunits as well as mutant forms of the MalK subunit. Prior to measuring nucleotide binding, we examined membrane preparations for the presence of each subunit from strains that contained all possible permutations of the three structural genes, malF, malG, and malK. We found that when all three genes were present or when malF and malK were present together, the corresponding antigens were detected easily on Western immunoblots and were soluble in the non-ionic detergent, Triton X-100. In contrast, all other permutations resulted in decreased amounts of antigen or antigen that was Triton X-100-insoluble. We relied on photocross-linking with 8-azido-[32P]ATP and ATP hydrolysis as indicators of the ability of the transport complex to interact with purine nucleotides. 8-Azido-[32P]ATP was photocross-linked to the MalK subunit. Photolabeling of MalK was inhibited by ATP, ADP, and GTP and not by other nucleotides. Photolabeling of MalK required the presence of MalF but not MalG. Mutations in malK that affect amino acid residues thought to be directly involved in nucleotide binding did indeed abolish labeling and resulted in loss of transport activity without affecting protein stability. In general, ATP hydrolysis correlated with the photocross-linking. A notable exception is the MalK941 mutant protein which retained the ability to be labeled by 8-azido-[32P]ATP but was unable to catalyze detectable levels of ATP hydrolysis. Some, but not all, of the malK mutations were dominant to wild type. To study the mechanism of dominance we devised a means of measuring the ability of different wild-type and mutant MalK proteins to interact with the MalF and MalG subunits. This assay relies on the fact that, when a bifunctional MalK-LacZ hybrid protein is associated with the MalF and MalG subunits, it is membrane-bound. Excess MalK competed with the MalK-LacZ hybrid protein for sites in the membrane and resulted in the hybrid fractionating as a soluble protein.(ABSTRACT TRUNCATED AT 400 WORDS)

ATP-Binding Cassette Transporters

Diagnostic accuracy of AIDS-related CNS lesions.

Because of the high incidence of neurological complications seen in patients with acquired immunodeficiency syndrome (AIDS), an increasing number of these cases are being referred to neurosurgeons for consideration of intracranial biopsy. To better determine the need for biopsy in these patients we evaluated the accuracy of non-tissue-based neurological diagnoses in AIDS patients who subsequently had a final diagnosis on the basis of biopsy or postmortem brain examinations. The records of 56 AIDS patients who had undergone either autopsy or brain biopsy were retrospectively reviewed. Of the ten patients who underwent biopsy, three were found to have a lesion that was different from the suspected diagnosis and that resulted in a change in treatment. Thirty patients with neurological symptoms had postmortem brain examinations. In the case of the 12 patients who had carried specific diagnoses and received treatments based on those diagnoses, only six diagnoses (50%) were proven correct at autopsy. Of the 18 cases that did not record a specific antemortem diagnosis, in only 5 were normal brains reported, while the others reported a variety of nonspecific or infectious findings. Twelve patients without neurological symptoms had postmortem brain examinations and only six of these (50%) had normal or slightly atrophic brains. Case reports of the others noted nonspecific findings most of which were suggestive of subacute HIV encephalitis. The poor rate of diagnostic accuracy in this series suggests that biopsy should be considered for atypical lesions or those that do not respond to empiric therapy. The use and relative sensitivities of various diagnostic studies are also discussed.

Acquired Immunodeficiency Syndrome

Actinic prurigo.

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Conjunctival Diseases

[A giant hepatic hemangioma and pregnancy].

We present a clinical case of a pregnancy with giant hepatic hemangioma. At term it presented like a consumption coagulopathy with thrombocytopenia. Maternal and fetal color Doppler was normal. The perinatal outcome was successful.

Adult

The activities of the Escherichia coli MalK protein in maltose transport, regulation, and inducer exclusion can be separated by mutations.

The maltose regulon consists of several genes encoding proteins involved in the uptake and utilization of maltose and maltodextrins. Five proteins make up a periplasmic binding-protein-dependent active transport system. One of these proteins, MalK, contains an ATP-binding site and is thought to couple the hydrolysis of ATP to the accumulation of substrate. Beside its function in transport, MalK has two additional roles: (i) it negatively regulates mal regulon expression and (ii) it serves as the target for regulation of transport activity by enzyme IIIGlc of the phosphotransferase system. To determine whether the three functions of MalK are separable, we have isolated and characterized three classes of malK mutations. The first type (class I) exhibited constitutive mal gene expression but still allowed normal transport of maltose; the second type (class II) lacked the ability to transport maltose but retained the ability to repress the mal genes. Class I mutations were localized in the last third of the gene, at amino acids 267 (Trp to Gly) and 346 (Gly to Ser). Mutations of class II were found at the positions 137 (Gly to Ala), 140 (delta Gln Arg), and 158 (Asp to Asn). These mutations are near or within the region of MalK that exhibits extensive homology to the B site of an ATP-binding fold. In addition, site-directed mutagenesis was used to add or remove one amino acid in the A site of the ATP-binding fold. Plasmids carrying these mutations also behaved as class II mutants. The third class of malK mutations resulted in resistance to the enzyme IIIGlc-mediated inhibitory effects of alpha-methylglucoside. These mutations did not interfere with the regulatory function of MalK. One of these mutations (exchanging a serine at position 282 for leucine) is located in a short stretch of amino acids that exhibits homology to a sequence in the Escherichia coli Lac permease in which alpha-methylglucoside-resistant mutations have been found.

ATP-Binding Cassette Transporters