Use of chemical clamps in denaturing gradient gel electrophoresis: application in the detection of the most frequent Mediterranean beta-thalassemic mutations.
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Base Sequence↗
Biomedical subjects
E Fernandez
Publications and source records attributed to E Fernandez.
Progressive feline pressure-overload: noninvasive assessment correlates with abnormalities in single cells.
Serial echocardiography and Doppler were used to monitor the progression of pressure-overload produced by banding the ascending aortas of young cats. The peak Doppler gradient across the band increased (as the animals grew in size) from 42 +/- 4.2 mmHg at 1 wk to 78 +/- 4.5 mmHg at 2-3 mos. Echocardiographic measurements of septal wall thickness increased significantly at 1 wk. Global ventricular function was unaltered in banded cats versus shams at each time point. However, in the subgroup of animals with an aortic-constricted area of < 0.025 cm2 at 1 wk, fractional shortening decreased by 40% at 2-3 mos. Contractile abnormalities were present in isolated myocytes from hypertrophied hearts. Mechanical function was more profoundly depressed in cells from hearts with echocardiographic evidence of ventricular decompensation. Echocardiographic and Doppler studies assessed cardiac size and function and identified indexes predictive of global and cellular myocardial dysfunction. The use of noninvasive techniques as a predictor of failure makes the feline model of progressive left ventricular pressure-overload useful for studies of cellular and molecular factors regulating not only the development of cardiac hypertrophy but also the transition from compensated hypertrophy to myocardial failure.
Analysis of Variance↗
Net uptake of lactate by rabbit hindlimb during hypoxia.
Lactate uptake by normoxic tissues is a pH-dependent phenomenon that occurs with increases in arterial lactate concentration. In this study we sought to determine the effect of hypoxia on lactate uptake by the rabbit hindlimb at different arterial blood pH (pHa). Two groups of rabbits were subjected to 20 min of arterial hypoxemia (PaO2 approximately 20 mm Hg). One group was allowed to maintain normal pHa (approximately 7.40 to 7.45; n = 5), whereas the other group was hyperventilated, resulting in hypocarbia and alkalemia (pHa > 7.50; n = 5). Lactate uptake was computed from measurements of femoral blood flow and the arterio-venous lactate difference. We found decreases in hindlimb O2 consumption during hypoxemia from 0.79 +/- 0.08 to 0.33 +/- 0.06 ml/min in the normal pH group (p < 0.01) and from 0.49 +/- 0.06 to 0.24 +/- 0.02 ml/min in the high pH group (p < 0.01). At that time there was net uptake of lactate by the hindlimb of the normal pH group (8.1 +/- 2.0 mumol/min; p < 0.001) and lactate release by the alkalemic group (1.61 +/- 1.0 mumol/min). Furthermore, hindlimb lactate uptake was associated with increased glucose consumption (p < 0.001). We hypothesize that the mechanisms responsible for lactate uptake by resting skeletal muscle during hypoxia may be similar to those that regulate lactate uptake during normoxic exercise.
Animals↗
Valproate-induced coma: case report and literature review.
OBJECTIVE: To report a case of hyperammonemia without hepatic dysfunction as a possible cause of lethargy, stupor, and coma in a woman after valproic acid (VPA) administration, and discuss the possible different mechanisms of ammonia elevation and coma. CASE SUMMARY: A woman diagnosed with complex partial seizures that secondarily generalize was treated with phenytoin (PHT) 250 mg/d for 18 years. Three months before admission, this dosage was increased to 300 mg/d and phenobarbital (PB) 100 mg/d was added because the seizures were incompletely controlled. The patient developed a progressive inability to walk. She was diagnosed as having PHT intoxication. VPA therapy was begun while PHT was being tapered and progressive impairment of consciousness occurred. This evolved into a coma without focal neurologic signs, and was accompanied by isolated hyperammonemia without hepatic failure. DISCUSSION: Adverse effects attributable to VPA were reviewed in the literature. Occasionally, VPA may lead to severe secondary effects such as hepatic failure and coma. In these cases increased blood concentrations of transaminases, bilirubin, and ammonia have been found. Several reports have stressed the existence of hyperammonemic coma without biochemical evidence of hepatic failure, which is what occurred in our patient. This suggests that isolated hyperammonemia and hepatic failure after VPA treatment may have a different biochemical basis. CONCLUSIONS: VPA-induced coma with hyperammonemia and without evidence of hepatic failure should be considered in patients being treated with PHT or PB when VPA is administered concomitantly. This case report shows the importance of clinical monitoring and immediate drug discontinuation when drowsiness, gastrointestinal symptoms, or lethargy occur.
Ammonia↗
A method for selective intracellular labeling of immunostained neurons in turtle retina.
We describe a method for direct intracellular staining under visual control of immunolabeled neurons in the turtle retina. Substance P was the antiserum used. It labels two different sizes of ganglion cells in turtle retina. Intracellular labeling under visual control was achieved by iontophoresis of Lucifer yellow or Neurobiotin. The best immunolabeling of substance P-immunoreactive (SP-IR) ganglion cells occurred after either Triton X-100 or freeze-thaw techniques to get good penetration of the antisera. However, this inevitably resulted in leaky cells and inadequate morphology of the ganglion cells subsequently stained by Lucifer yellow and Neurobiotin. Most successful immunocytochemical labeling followed by intracellular labeling was achieved with light fixation (15 min in 4% paraformaldehyde) and long incubation time in the primary antiserum (4 days). Before intracellular labeling, dendritic tree shape, dendritic field size, and stratification of SP-IR ganglion cells were not sufficiently revealed for correct classification of these cells. After the selective intracellular staining described here, we were able to identify and characterize one of the populations of substance P-IR ganglion cells types as large-field, monostratified G20 ganglion cells.
Adaptation, Ocular↗
Spinal cord transection in adult rats: effects of local infusion of nerve growth factor on the corticospinal tract axons.
The spinal cord of adult female rats was completely transected at the T8 level. Nerve growth factor (NGF) was administered at the lesion site via indwelling, implanted, osmotic minipumps. Purified NGF was supplied at doses of 100, 200, and 500 micrograms during a 30-day period. Control rats were treated with saline. At the end of the treatment, the proximal stump of corticospinal tract axons in the spinal cord was labeled with anterograde transported horseradish peroxidase (HRP) injected into the sensorimotor cortex. In control rats, the corticospinal tract axons ended abruptly, proximal to the zone of maximal damage. Sterile swellings developed at the axon tips, and no labeled axonal sprouts were apparent. On the contrary, in NGF-treated animals, the leading front of the corticospinal tract axons showed a trend of approaching the zone of maximal damage following abnormal paths through the dorsal-injured white matter. Axonal sprouts were seen more proximally, traveling toward the transection site in aberrantly located dorsal paths, completely outside the normal position of the corticospinal tract. NGF seems to partly restore the pattern of the regenerative behavior of the severed corticospinal tract axons after spinal cord transection in newborn rats, i.e., the induction of axonal sprouting in aberrantly located dorsal paths. An automated image analysis of the HRP reaction field close to the transection site demonstrated that the density of HRP-labeled axons in the corticospinal tract was significantly higher in the NGF-treated rats than in the control rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
The influence of L-acetylcarnitine on reinnervation of the oculomotor nerve.
In guinea-pigs the oral administration of L-acetylcarnitine (L-AC) markedly favours the process of reinnervation of the oculomotor nerve sectioned at intracranial level. The gains of the horizontal and vertical vestibulo-ocular reflexes (HVOR, VVOR) were taken into consideration in testing the functional recovery of the nerve. As a consequence of the drug administration, 24 weeks after the operation the gains of the treated animals were higher than those of the controls. Reduction of misalignments of the stimulus-response orientation was also observed in treated animals as compared to the controls. This suggests that L-AC potentiates motor reinnervation by enhancing the nerve-growing processes and favouring a better consolidation of the appropriate neuromuscular synapses. The increased gain, and the improvement of the alignment in ocular responses, due to L-AC would allow for an increase of visual function during head movement by optimizing gaze stability.
Acetylcarnitine↗
Endemic HTLV-II infection among Tobas and Matacos Amerindians from north Argentina.
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Adolescent↗
Antinociceptive effects of Ca2+ channel blockers.
The antinociceptive action of four Ca2+ channel blockers, nifedipine, nimodipine, verapamil and diltiazem, was evaluated and compared to that of morphine using three algesiometric tests in mice and rats, namely, formalin, writhing and modified hot-plate test. Dose-response curves for all the drugs tested were similar and a significant dose-dependent antinociceptive action was evident in the formalin and writhing tests. However, in the hot-plate test, only nimodipine exhibited a significant analgesic effect, confirming the misleading results previously reported for this test. The findings suggest a pharmacological role of Ca2+ channel blockers in the modulation of antinociception under acute conditions. The analgesic action of Ca2+ channel blockers could be mediated by an increase in the nociceptive threshold resulting from interference with Ca2+ influx at opioid receptors, because Ca2+ influx is critical for the release of neurotransmitters and other substances implicated in nociception and inflammation. It is suggested that if a substance has a Ca2+ channel blocking effect, it should probably have some antinociceptive properties.
Analgesics↗
Effect of ethanol on the Na(+)- and the Na+,K(+)-ATPase activities of basolateral plasma membranes of kidney proximal tubular cells.
The Na(+)- and the Na+,K(+)-ATPase activities of basolateral plasma membranes from rat kidney proximal tubular cells were affected differentially by ethanol. Moreover, at concentrations of ethanol that can be reached in vivo in the blood plasma (50 mM) there was a significant effect on the Na(+)-ATPase activity and practically no effect on the Na+,K(+)-ATPase activity.
Adenosine Triphosphatases↗
Giant axonal neuropathy: report on a case with focal fiber loss.
We report on a 5 1/2 year-old boy with chronic progressive polyneuropathy, ataxia, and pyramidal signs. His hair was not curled. Sural nerve biopsy disclosed many axons enlarged by accumulation of 10-nm neurofilaments and a marked variability in the number of myelinated fibers as well as in the amount of axonal enlargements among different fascicles. These findings and the electrophysiological data were consistent with a giant axonal polyneuropathy with a multifocal fiber loss.
Axons↗
Tris discriminates between the different alpha-glucosidase activities from extracts of human neutrophils.
In an attempt to detect acid maltase deficiency in neutrophils from patients with type II glycogenosis, without interference from the 'renal' alpha-glucosidase activity present in these cells, we have evaluated the contribution of the renal component in the total activity measured at pH 4.0 in extracts of human neutrophils. The renal contribution is about 13-25% and renal glucosidase appears to be closely related to the enzyme present on the epithelium of small intestine, which is known to be inhibited by Tris. We have used this compound as a selective inhibitor of the renal component of alpha-glucosidase activity measured at pH 4.0 in total extracts of neutrophils. Our results demonstrate that 0.1 mol/L Tris is an inhibitor of the renal alpha-glucosidase present in neutrophils and can be used to reduce the interference from this enzyme in assays of acid maltase.
Glycoside Hydrolase Inhibitors↗
Sensory and affective components of pain: separation and synthesis.
It has become increasingly accepted that pain is not simply a sensation generated by nociceptors, but a perceptual phenomenon with particular emotional qualities. The purpose of this article is to bring together vastly different streams of research on the divisibility of pain into sensory and affective components. Empirical evidence for this divisibility is drawn from recent studies using multivariate statistics, signal detection theory, and unidimensional scaling. An important conclusion is that separable though pain components may be, they are not necessarily independent. In critiquing previous research, new criteria are derived for partitioning pain into sensory and affective components. Finally, speculations are offered as to how these same components might be synthesized on the basis of theories of perceptual organization.
Affective Symptoms↗
Epidemiology of prostatic disorders in the city of Barcelona.
Although disorders of the prostate are among the most prevalent problems in elderly males, little is known about their epidemiological characteristics. The 1986 Barcelona Health Interview Survey (BHIS), a cross-sectional study of a random sample of the noninstitutionalized population of the city, was used to estimate the prevalence of prostatic disorders (PD) and to analyse sociodemographic characteristics, self-perceived health, and use of health services among males reporting PD. Ninety-eight out of 1218 males over 45 years old (8%) reported having PD, the prevalence increasing substantially with age. A pattern of increasing prevalence with decreasing occupational class was found. As compared to subjects not reporting PD, individuals reporting PD reported slightly more restricted activity days (prevalence odds ratio [POR] = 1.28; 95% confidence interval [CI]: 0.52-2.80), more chronic disorders (POR = 4.75; 95% CI: 2.04-13.53), and worse self-perceived health (POR = 1.55; 95% CI: 0.92-2.58). Medical visits and hospitalizations were also higher in the prostatic group (P < 0.05). Men reporting PD appear to constitute a subgroup with increased morbidity and health services use.
Age Factors↗
Role of tissue hypoxia as the mechanism of lactic acidosis during E. coli endotoxemia.
We compared the hemodynamic and metabolic alterations produced in rabbits by similar decreases in cardiac output created by inflating a balloon placed in the right ventricle (n = 6) with those produced by an intravenous bolus of Escherichia coli lipopolysaccharide (LPS; SEP group; n = 6). We measured O2 consumption (VO2), O2 transport (TO2), and O2 extraction ratio (ERO2) for the whole animal and also for the left hindlimb. Both groups experienced similar decreases in cardiac output, systemic TO2, and VO2 and similar increases in ERO2. For the hindlimb, TO2 was similar, but VO2 and ERO2 were lower for the SEP group 30 min after LPS administration (P less than 0.05); however, this difference disappeared during the remainder of the experiment. Arterial lactate concentration was greater (P less than 0.05) for the SEP group. There were no differences in skeletal muscle PO2, measured with a multiwire surface electrode, or in cardiac and skeletal muscle concentrations of high-energy phosphates. We hypothesize that a direct effect of LPS on cellular metabolism may have resulted in greater arterial lactate concentration for the SEP group.
Acidosis, Lactic↗
Reconstruction of peripheral nerves: the phenomenon of bilateral reinnervation of muscles originally innervated by unilateral motoneurons.
It is well known that after reconstruction of sectioned peripheral nerves in adult mammals, denervated muscles are reinnervated by the axotomized motoneurons lying in the original motonucleus. It is less well known that these muscles can also be reinnervated by uninjured motoneurons lying in the homologous contralateral motonucleus. Therefore, after nerve reconstruction, bilateral motoneuron reinnervation of muscles can occur. Contralateral motoneurons sprout axons that cross the midline, grow in the reconstructed nerve, and reach muscle targets. This phenomenon was observed after reconstruction of several different peripheral nerves in adult mammals, including the oculomotor nerve in guinea pigs and the facial and sciatic nerves in rats. The retrograde axonal transport of horseradish peroxidase was used for the study of the organization of the brainstem and spinal cord motonuclei. Horseradish peroxidase was injected into the medial rectus muscle, the stylohyoid muscle, and the trunk of the sciatic nerve. The distance between the homologous motonuclei of both sides influenced the occurrence of this phenomenon. In fact, bilateral reinnervation of muscles after nerve reconstruction was found in 36% (sciatic nerve), 50% (facial nerve), and 100% (oculomotor nerve) of the operated animals. The total number of contralateral motoneurons found were 14% (oculomotor nerve), 8% (facial nerve), and 5% (sciatic nerve). Bilateral reinnervation of muscles was evoked by both immediate and delayed peripheral nerve repair and was a stable phenomenon, seen between 3 and 21 months after facial nerve reconstruction.
Animals↗
Cholinergic receptors in the human vas deferens.
This study represents the first investigation demonstrating the contractile response to exogenous acetylcholine (ACh) in the isolated human vas deferens. Pharmacological characterization of cholinergic receptors was achieved using selective antagonists to define receptor subtypes. In the HVD the effect of exogenous ACh is revealed as a dose-dependent sudden increase in the basal tension of the vasa. The ACh receptors of the HVD were competitively antagonized by atropine (ATR) with a high pA2 value (8.78). The main finding of this study is the presence of cholinergic receptors of the pharmacologically defined M1-ACh subtype in the isolated HVD, according to the pA2 values obtained with pirenzepine (PRZ) 7.39, AF-DX 116 (AF) 5.92 and 4-DAMP 5.65, M1-ACh, M2-ACh and M3-ACh selective antagonists, respectively. Prazosin (PZ), a selective alpha 1-adrenergic antagonist, displayed a similar competitive antagonism for the contractile response evoked both by ACh (pA2 = 8.69) and NE (pA2 = 8.58) in the HVD. The antagonism exerted by PZ on the ACh-induced contractile response of the HVD, suggests that ACh probably acts at a presynaptic level stimulating the release of NE from an adrenergic neuron. According to these findings, the receptor involved in this action, located in the proximity of the nerve terminals, seems to be of the M1-ACh subtype.
Acetylcholine↗
Experimental repair of the oculomotor nerve: the anatomical paradigms of functional regeneration.
In adult guinea pigs, the oculomotor nerve was sectioned proximally (at the tentorial edge) or more distally (at the orbital fissure) and immediately repaired by reapproximation. During a 24-week postoperative period, extrinsic eye motility was assessed by analyzing the vestibulo-ocular reflexes. The regenerated oculomotor nerve was studied morphometrically on semi-thin histological sections at 16 and 24 weeks postinjury. The selectivity of muscle reinnervation was investigated by injection of both single (horseradish peroxidase) and double (fluorescent dyes) retrograde axonal tracers into the eye muscles. Following proximal repair of the oculomotor nerve, the degree of recovery of extraocular motility varied among different animals and remained stable over long-term observations. In animals with poor recovery, aberrant eye movements were always found, and the somatotopic map of the reinnervated eye muscles was greatly altered. Distortions of the central representation were also seen in those animals in which a good level of functional recovery was seen. However, in animals with good recovery, a topographic bias was re-established by about 65% of the original neuronal population, as opposed to 26% in the animals with poor recovery. Neurons located contralateral to the axotomized nucleus sprouted intra-axially and projected their axons to denervated eye muscles. The number and diameter of the regenerated axons, the number and soma diameter of the axotomized neurons, and the ratio of distal axonal branches to proximal supporting neurons were all related to the degree of functional recovery. Following repair of the oculomotor nerve at the orbital fissure, extraocular motility had recovered in all of the animals at 16 weeks without aberrant phenomena. Functional regeneration of the distally transected oculomotor nerve is thought to be the result of selective muscle reinnervation.
Animals↗