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Biomedical subjects

A M Fernandez

Publications and source records attributed to A M Fernandez.

29 records · Page 2Linked to original sources

Chemical heterogeneity in adult rat cerebellar Purkinje cells as revealed by zebrin I and low-affinity nerve growth factor receptor immunocytochemical expression following injury.

Cerebellar Purkinje cells in rat express low-affinity nerve growth factor receptor during development, but rarely in normal adult animals. However, after either mechanical injury or colchicine treatment during adulthood, these cells re-express low-affinity nerve growth factor receptor-immunoreactive protein. Two Purkinje cell subpopulations were defined in normal adult cerebellum by the presence or the absence of zebrin I antigen. Nevertheless, it remains an open question as to whether low-affinity nerve growth factor receptor-immunoreactive protein can be expressed by all damaged Purkinje cells, independent of their location and their staining with antibodies against intrinsic molecular markers that reveal Purkinje cell heterogeneity, such as zebrin I. In this study, a serial-section immunocytochemical mapping of the expression zebrin I and low-affinity nerve growth factor receptor, using specific monoclonal antibodies, we carried out in colchicine-treated rats. After mechanical damage of the cerebellar cortex, co-localization of these antigens at the cellular level was also analysed in thin adjacent sections, and by using a combined immunocytochemical staining method in individual sections. The findings revealed the existence of three sub-sets of Purkinje cells: (1) two complementary groups distinctly immunoreactive to one antibody, but not to the other and (2) a third group that contained double-labelled cells. In contrast, co-expression of both antigens was never observed following mechanical lesions. The seemingly independent response to mechanical injury of Purkinje cells located in different zebrin-defined compartments, indicates that particular subpopulations of Purkinje cells may respond differentially to traumatic injury.

Animals↗

A role for cholesterol as a structural effector of the nicotinic acetylcholine receptor.

The effects of cholesterol on the protein structure and on the ionic channel activity of purified acetylcholine receptor (AcChR) reconstituted into lipid vesicles have been studied, respectively, by Fourier-transform infrared spectroscopy and by rapid kinetics of cation influx. Reconstitution of the AcChR in asolectin phospholipid vesicles in the absence of either cholesterol or the nonpolar lipids present in crude asolectin extracts results in a considerable loss of the ability of the AcChR to support cation channel function. This functional loss is accompanied by spectral changes in the conformationally-sensitive amide I band of the protein infrared spectrum which are indicative of alteration in the protein secondary structure. Quantitative estimation of such alteration by band-fitting analysis reveals a marked decrease in ordered protein structures such as the alpha-helix and beta-pleated sheet, concomitant with an increase in less ordered structures appearing at 1644 cm-1 in the infrared spectrum. Furthermore, the addition of increasing amounts of cholesterol to the reconstituted bilayer produces a progressive, complete recovery both in the control of cation channel function and in the infrared spectrum. This restoration of AcChR structure and function by cholesterol, however, does not occur when the AcChR is reconstituted in vesicles made from purified egg phosphatidylcholine, thus suggesting that the presence in the reconstituted bilayer of phospholipids other than phosphatidylcholine may be required for cholesterol to exert its modulatory effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Amides↗

Posterior interosseous nerve entrapment in rheumatoid arthritis.

Posterior interosseous nerve (PIN) entrapment is a rare complication of rheumatoid arthritis (RA) which, together with extensor tendon rupture and metacarpophangeal joint dislocation, should be considered in the differential diagnosis of inability to extend the fingers. The inability to extend the thumb in PIN entrapment is a useful distinguishing clue on physical examination, and nerve conduction studies confirm the diagnosis. Elbow joint swelling and compression of the PIN at the arcade of Frohse are the main reasons for PIN entrapment in RA. Intraarticular steroid injections and surgical intervention resolve symptoms of PIN entrapment in RA. In our case, the addition of methotrexate, which induces a rapid antiinflammatory effect, resulted in resolution of weakness with complete recovery in the extensor muscles of the fingers.

Arthritis, Rheumatoid↗

Recurrent leukocytoclastic vasculitis as the initial manifestation of acute myelomonocytic leukemia.

The association between cutaneous and systemic leukocytoclastic vasculitis and myeloproliferative and lymphoproliferative malignancies has been reported. We describe a patient who developed recurrent episodes of leukocytoclastic vasculitis 10 months before the diagnosis of acute myelomonocytic leukemia. To our knowledge, this is the first documented case of leukocytoclastic vasculitis presenting as the initial manifestation of acute myelomonocytic leukemia, a subtype of acute myelogenous leukemia.

Female↗

Long-term trends in blood lead levels among children in Chicago: relationship to air lead levels.

OBJECTIVES: To evaluate trends in blood lead levels among children in Chicago from 1968 through 1988, and to determine the impact of the changes in the Centers for Disease Control and Prevention (CDC) blood lead level of concern. METHODS: We reviewed a systematic sample of blood lead screening records of the Chicago Department of Health Laboratory for high-risk children aged 6 months to 5 years. Median blood lead levels for each quarter of the years 1974 through 1988 were determined and regressed against mean air lead levels recorded at air-monitoring stations in Chicago during the same period. RESULTS: Median blood lead levels declined from 30 micrograms/dL in 1968 to 12 micrograms/dL in 1988, and were strongly associated with declining average air lead levels (r = .8, P < .001) from 1974 through 1988. A regression model using log-transformed data predicted a decline of 0.56 microgram/dL in the median blood lead level with each 0.1 microgram/m3 decline in the mean air lead level when the air lead level was near 1.0 microgram/m3; the predicted slope was steeper at lower air lead levels. Despite the nearly 20-fold reduction in air lead levels, the median blood lead level of 12 micrograms/dL in 1988 indicates substantial continuing lead exposure. The CDC blood lead level of concern was lowered twice from 1968 to 1988, but due to the decline in blood lead levels, fewer than 30% of the children were above the level of concern throughout most of the study. CONCLUSION: Although substantial lead exposure persists in Chicago, reductions in airborne lead emissions seem to have contributed to a long-term decline in the median blood lead level of high-risk Chicago children.

Age Factors↗

Labeling of the nicotinic acetylcholine receptor by a photoactivatable steroid probe: effects of cholesterol and cholinergic ligands.

A photoactivatable steroid, p-azidophenacyl 3 alpha-hydroxy-5 beta-cholan-24- ate (APL), has been synthesized and used instead of cholesterol to functionally reconstitute purified acetylcholine receptor (AcChR) into vesicles made of asolectin phospholipids. Upon irradiation, the extent of AcChR photolabeling by APL is directly proportional to the amount of APL incorporated into the reconstituted vesicles and the maximum stoichiometry observed corresponds to approx. 50 mol of APL bound per mol of AcChR. Furthermore, all four subunits of the AcChR become labeled by APL and the observed labeling pattern resembles the 2:1:1:1 stoichiometry characteristic of these subunits within the AcChR complex. The presence of either cholesterol or neutral lipids from asolectin in the reconstituted bilayer decreases both, the incorporation of APl into the vesicles and the covalent labeling of the AcChR upon irradiation, without altering the stoichiometry of labeling in AcChR subunits stated above. This suggests that the potential interaction sites for the photoactivatable probe in the reconstituted AcChR are mostly those normally occupied by the natural neutral lipids. Carbamylcholine, a cholinergic agonist, also reduces the extent of APL photolabeling of the AcChR in a dose-dependent manner but, in contrast to the effects of cholesterol, the presence of carbamylcholine alters the stoichiometry of labeling in the AcChR subunits. This, along with the observation that such a decrease in the extent of APL photolabeling caused by carbamylcholine can be blocked by preincubation with alpha-bungarotoxin, suggest that AcChR desensitization induced by prolonged exposure to cholinergic agonists encompasses a rearrangement of transmembrane portions of the AcChR protein, which can be sensed by the photoactivatable probe. Conversely, presence of (+)-tubocurarine, a competitive cholinergic antagonist, has no effects on altering either the extent of APL photolabeling of the AcChR or the distribution of the labeling among AcChR subunits.

Affinity Labels↗

Protein structural effects of agonist binding to the nicotinic acetylcholine receptor.

The effects on the protein structure produced by binding of cholinergic agonists to purified acetylcholine receptor (AcChR) reconstituted into lipid vesicles, has been studied by Fourier-transform infrared spectroscopy and differential scanning calorimetry. Spectral changes in the conformationally sensitive amide I infrared band indicates that the exposure of the AcChR to the agonist carbamylcholine, under conditions which drive the AcChR into the desensitized state, produces alterations in the protein secondary structure. Quantitative estimation of these agonist-induced alterations by band-fitting analysis of the amide I spectral band reveals no appreciable changes in the percent of alpha-helix, but a decrease in beta-sheet structure, concomitant with an increase in less ordered structures. Additionally, agonist binding results in a concentration-dependent increase in the protein thermal stability, as indicated by the temperature dependence of the protein infrared spectrum and by calorimetric analysis, which further suggest that AcChR desensitization induced by the cholinergic agonist implies significant rearrangements in the protein structure.

Animals↗

Axonal and non-axonal immunolocalization of cytosolic aspartate aminotransferase (cAATase), GABA and glutamic acid decarboxylase (GAD) in the rat cochlear nucleus.

The localization of both cAATase activity, by histoenzymological method, and the immunoreactivity against cAATase, were investigated in the cochlear nucleus of rats. The immunohistochemical determination of cAATase was carried out using the PAP method, with an antiserum obtained from rabbits immunized with porcine cAATase. It was also studied both the immunolocalization of GABA-like and GAD-like substances. Our observations, with light microscope, revealed weak cAATase activity in the small neurons, and a more intense one in the fibers surrounding neuronal bodies. The large neurons presented a very weakly activity within their neuronal bodies and dendrites, but it was strongly found in granulations that surround the perikaryon and dendrites. cAATase immunoreactivity presents the same distribution as the enzymological activity. In the same way, we have investigated the pattern of distribution of both GABA- and GAD-like substances. Immunolocalization of these substances was similar to that found for cAATase. In the control sections incubated with Gostatin (0.05 mM), cAATase activity was absent. The immunoreactivity was also negative in every immunohistochemical control sections. These facts suggest that aspartate could intervene as a co-neurotransmitter or neuromodulator in the rat cochlear nucleus, and that axonic endings could contain cAATase, GABA and GAD. It was also found immunoreactivity against cAATase, GABA and GAD, in neuronal bodies, dendrites and glial processes, in close association with capillary wall. These observations have led us to suggest the possible co-localization and co-release of both GABA and aspartate from synaptic and non-synaptic sites in the cochlear nucleus.

Animals↗

Progressive cellular immune impairment leading to development of AIDS: two-year prospective study of HIV infection in drug addicts.

We have studied immunological, serological and clinical abnormalities in 264 HIV-positive and HIV-negative drug abusers. Ninety percent of the 264 drug addicts (mean age 26 +/- 0.8 years) were found to have HIV antibodies and there was a significant increase (P less than 0.01) in the frequency of HIV antibody positivity with increasing duration of exposure to parenteral drug abuse. There was a very strong correlation between the progressive decline of the mean T4+ helper/inducer cells and T4+/T8+ ratio, the low response to pokeweed mitogen and the more severe clinical manifestations of HIV infection. Impairment of delayed-type hypersensitivity reaction to recall antigens was only seen in group IV as defined by the Center for Disease Control. Within group IV, anergy was found to be highly associated (83%) in patients with opportunistic infections. All other HIV-positive addicts from groups II and III, as well as HIV-negative addicts had normal in vivo responses to test antigens. We have also analysed in a prospective follow-up lasting 6-24 months, the evolution of HIV infection in a cohort of 50 HIV-antibody-positive drug addicts. Thirty-two percent showed clinical progression and most of the drug addicts that proceeded to full-blown AIDS developed anergy (82%) prior to clinical deterioration with development of opportunistic infections. We conclude, that in seropositive drug addicts a low absolute count of helper/inducer cells (mean +/- s.e. = 243 +/- 48 cells/mm3), a low response to pokeweed mitogen and anergy are predictive markers of progression to AIDS.

Acquired Immunodeficiency Syndrome↗

Technology assessment in healthcare: a review and description of a "best practice" technology assessment process.

BACKGROUND: Technology assessment has become a rapidly growing component of the healthcare system. It has assumed a functional role in operational settings and is rapidly impacting decisions involving purchasing, coverage, and reimbursement. This review is intended to assist the healthcare decision maker in considering the application of technology assessment in healthcare, so as to maximize the efficiency of future purchasing decisions. METHODS: This "best practice" was synthesized after identifying key institutions performing technology assessment in healthcare and analyzing their working processes, including literature review, consensus panel discussions, and expert opinion. RESULTS: We describe this best practice on a reiterative loop that consists of five processes: awareness, strategic appropriateness, analysis versus need, acquisition and implementation, and reassessment. Typical barriers to adoption of technology assessment are also identified and discussed. CONCLUSIONS: This review suggests a common terminology for the core processes involved in technology assessment, thereby facilitating a more uniform understanding among the different components of the healthcare system (i.e., payer, provider, and society) while recognizing their different perspectives.

Academies and Institutes↗