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Molecular determinants for modulation of persistent sodium current by G-protein betagamma subunits.

Voltage-gated sodium channels are responsible for the upstroke of the action potential in most excitable cells, and their fast inactivation is essential for controlling electrical signaling. In addition, a noninactivating, persistent component of sodium current, I(NaP), has been implicated in integrative functions of neurons including threshold for firing, neuronal bursting, and signal integration. G-protein betagamma subunits increase I(NaP), but the sodium channel subtypes that conduct I(NaP) and the target site(s) on the sodium channel molecule required for modulation by Gbetagamma are poorly defined. Here, we show that I(NaP) conducted by Na(v)1.1 and Na(v)1.2 channels (Na(v)1.1 > Na(v)1.2) is modulated by Gbetagamma; Na(v)1.4 and Na(v)1.5 channels produce smaller I(NaP) that is not regulated by Gbetagamma. These qualitative differences in modulation by Gbetagamma are determined by the transmembrane body of the sodium channels rather than their cytoplasmic C-terminal domains, which have been implicated previously in modulation by Gbetagamma. However, the C-terminal domains determine the quantitative extent of modulation of Na(v)1.2 channels by Gbetagamma. Studies of chimeric and truncated Na(v)1.2 channels identify molecular determinants that affect modulation of I(NaP) located between amino acid residue 1890 and the C terminus at residue 2005. The last 28 amino acid residues of the C terminus are sufficient to support modulation by Gbetagamma when attached to the proximal C-terminal domain. Our results further define the sodium channel subtypes that generate I(NaP) and identify crucial molecular determinants in the C-terminal domain required for modulation by Gbetagamma when attached to the transmembrane body of a responsive sodium channel.

Amino Acid Sequence↗

Conceptualizing and measuring integration: findings from the health systems integration study.

Given the pressures for health care reform, interest in the concept of integrated or organized delivery systems as a means to offer more coordinated cost-effective care is growing. This article has two primary objectives: (1) to clarify the different types of integration associated with the notion of an organized delivery system, and (2) to share the results from an ongoing study of 12 organized delivery systems. The findings indicate a moderate level of integration overall, particularly in the areas of culture, financial planning, and strategic planning. The study found that corporate staff respondents perceive their systems to be more integrated and effective than do operating unit managers, and that some functional integration areas are positively associated with both physician-system and clinical integration that, in turn, are positively related to each other. Overall, perceived integration was found to be positively associated with perceived effectiveness.

Administrative Personnel↗

The pentobarbital anesthetized dog: an animal model for assessing chemically induced changes in renal function and ultrastructure.

An experimental procedure was devised using the pentobarbital-anesthetized dog that could be used for the comprehensive evaluation of the renal effects of chemicals. After IV or renal arterial administration of 0.9% saline solution (vehicle), 12 renal function determinants were continuously monitored for periods of 2 and 6 hours. At the completion of the 2 or 6 hours of study, the kidneys of a number of dogs (usually between 1 and 7) in each vehicle-treated group were subjected to a modification of the intravascular perfusion-of-fixative technique to evaluate the ultrastructural status of the outer cortical, inner cortical, and outer medullary tissue. The remaining dogs (at least 3) in each vehicle-treated group were given a nonnephrotoxic, but maximally effective, diuretic dose of ethacrynic acid, which enabled an assessment of the functional integrity of the thick ascending limb of Henle's loop. Renal function and glomerular and tubular ultrastructure remained stable in the pentobarbital-anesthetized dog for up to 6 hours after administration of vehicle. Sustained infusion of inulin (included in the procedure to estimate glomerular filtration rate) throughout the duration of the experiments, and pentobarbital anesthesia of various durations did not alter the morphologic status of the canine nephron. The procedure used for the renal perfusion of fixative circumvented any manipulation of the kidneys before fixation and allowed for the acquisition of normal (unaltered) appearing tissue from all areas of the kidneys. The responses of pentobarbital-anesthetized dogs to ethacrynic acid administration were similar when given 2 and 6 hours after the vehicle administration.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia, General↗

[Spectral-correlation analysis of high-frequency EEG components of the rabbit cerebral cortex during orienting and defensive responses].

A change was shown in the EEG power spectra of the sensorimotor, visual and auditory neocortical areas and the dorsal hippocampus, and corresponding coherence functions in the range of 14.6-110 c/s during an orienting reaction and the formation of a defensive conditioned reflex in rabbits. It was suggested that the rhythm of about 5 c/s appearing during formation of the conditioned reflex, serves for the functional integration of the brain structures which is specific for a certain functional state.

Animals↗

Evaluation of cell membrane integrity of spermatozoa by hypoosmotic swelling test - "water test" in mice after intraperitoneal daidzein administration.

The functional integrity of the cell membrane of the mice sperm after daidzein administration intraperitoneal was assessed by studying the swelling reaction of the sperm when suspended in distilled water. The results obtained in water test indicate that daidzein, one of the phytoestrogens, decreases the number of normal and live sperm (swollen) in the samples obtaining from vas deferens. This effect was statistically significant in the mice after 7 and 14 days of daidzein administration. In mice treated with daidzein the number of dead (unswollen) sperm was increased, especially in animals after 14 days of experiment.

Animals↗

The development of drawing in children with congenital focal brain injury: evidence for limited functional recovery.

Children with pre- or perinatal injury to right hemisphere (RH) brain regions show impairment of spatial integrative functions similar to that observed among adults with comparable injury. Unlike adults, children show considerable improvement with development on a range of spatial construction tasks which require spatial integration. Such gains could reflect true recovery of spatial integrative abilities. Alternatively, the improvement could be more limited in scope, reflecting the development of compensatory strategies which are task specific and allow the children to circumvent, rather than overcome, their primary spatial disorders. The studies presented here examined this distinction within the context of drawing tasks in which the child was first asked to draw a house and then an impossible house. The impossible house task was designed to examine the extent to which children rely on graphic formulas in generating organized drawings. The results showed that while all of the children with RH injury make considerable progress in free drawing into the school age period, they are very reliant on the use of graphic formulas. When given a task which requires them to alter their drawings, they did not change the spatial configuration of the depicted object. Rather they found alternate ways to render the object 'impossible'.

Adolescent↗

[The pathological and the compensatory in cerebral ischemia: molecular aspects].

INTRODUCTION: An integral approach of the whole mechanisms involved during ischemic brain damage let us to realize that some phenomena, usually considered as 'pathological' make an integrate and harmonious response directed towards the preservation of neuronal survival and functional integrity of the whole system. DEVELOPMENT: It has been proposed that cellular lysis in a core of lesion may be protective for the surrounding tissue, as well as the development of vasoconstriction and cellular edema, in the face of the loss of intracellular calcium homeostasis. Some recent findings tend to support this hypothesis. Adaptative response to lower oxygen and glucose availability is linked with the inhibition of neuronal activity, and therefore, with their involved functions. Formation of prostanoids and hydrogen peroxide, and release of interleukin and neurotrophic factors are related to the expression of immediate early genes, the enhancement of cellular response during ischemic conditions and also related to the induction of mechanisms of repair and regeneration, where astroglial cells play a key role. CONCLUSIONS: Pharmacological inhibition, hypothermia and hyperbaric oxygenation had been employed with a relative success in the clinical management of ischemic injured patients. That interventions are in agreement with some of these endogenous compensatory mechanisms, which become the brain less vulnerable that it is generally considered. The effectiveness of therapeutics in cerebrovascular disease must be increased if this compensatory response is adequately directed by physicians.

Adaptation, Physiological↗

Integrity of the auditory pathway in young children with congenital and postmeningitic deafness.

This study compares the functional integrity of the auditory pathways of congenitally deaf and postmeningitically deaf children. We used the electrical auditory brain stem response evoked by promontory stimulation to assess 49 profoundly deaf children before cochlear implantation. The age at implantation ranged from 21 months to 15 years (mean 4.5 years). The onset of deafness was either congenital or up to the age of 2 years (mean 5 months). The cause of deafness was meningitis in 19 children (39%) and congenital in 30 (61%). The number of children with identifiable waveform components (eV, eIII, and eII) was significantly greater in the congenitally deaf group. We also analyzed the amplitudes, the latencies, and 4 parameters of the amplitude input-output functions. All the statistically significant differences were in favor of better responses in the congenitally deaf children. These results suggest that the functional status of the peripheral neurons of the auditory pathways may be more intact in congenitally deaf children than in postmeningitic children.

Adolescent↗

What is an organized infant?

Organization is a term used to describe the establishment of integrated functioning between an infant's physiologic and behavioral systems. When these systems are well integrated, the infant is able to interact with the environment without disruptions in autonomic, motor, and state functioning. As a result, he will not experience alterations in vital signs, color, state, or digestive functions that may compromise his future growth and development. Infants who can maintain or regain physiologic and behavioral organization will conserve energy for growth, provide caregivers with clear behavioral cues that indicate their needs, facilitate the establishment of parent-infant attachment, and use self-consoling or habituating behaviors when interacting with an environment that is too stimulating. As a result, developmental outcomes can be enhanced, particularly in infants who are at risk for neurobehavioral sequelae and developmental delays. Nurses can help infants achieve and maintain organization by providing an environment that supports their physiologic and behavioral organization.

Adaptation, Psychological↗

A genetic analysis of glutamatergic function in Drosophila.

Neurotransmitters are essential for communication between neurons and hence are vital in the overall integrative functioning of the nervous system. Previous work on acetylcholine metabolism in the fruit fly, Drosophila melanogaster, has also raised the possibility that transmitter metabolism may play a prominent role in either the achievement or maintenance of the normal structure of the central nervous system in this species. Unfortunately, acetylcholine is rather poorly characterized as a neurotransmitter in Drosophila; consequently, we have begun an analysis of the role of glutamate (probably the best characterized transmitter in this organism) in the formation and/or maintenance of nervous system structure. We present here the results of a series of preliminary analyses. To suggest where glutamatergic function may be localized, an examination of the spatial distribution of high affinity [3H]-glutamate binding sites are presented. We present the results of an analysis of the spatial and temporal distribution of enzymatic activities thought to be important in the regulation of transmitter-glutamate pools (i.e., glutamate oxaloacetic transaminase, glutaminase, and glutamate dehydrogenase). To begin to examine whether mutations in any of these functions are capable of affecting glutamatergic activity, we present the results of an initial genetic analysis of one enzymatic function, glutamate oxaloacetic transaminase (GOT), chosen because of its differential distribution within the adult central nervous system and musculature.

Animals↗

Cryopreserved rat liver slices: a critical evaluation of cell viability, histological integrity, and drug-metabolizing enzymes.

The effects of a cryopreservation procedure on the biochemical, morphological and functional integrity of rat liver slices just after thawing and after 24 h culture were evaluated. Freshly prepared slices were incubated in modified University of Wisconsin solution containing 50% fetal calf serum and 10% dimethyl sulfoxide for 20 min on ice prior to a rapid cooling in liquid nitrogen. After 10-40 days, slices were thawed rapidly at 42 degrees C. Total protein content and (3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) (MTT) reduction were well preserved at thawing, whereas ATP content was markedly decreased relative to freshly prepared slices (-83%). The major microscopic findings in sections of just-thawed liver slices consisted of hepatocellular dissociation and minimal apoptosis. The qualitative profile of antipyrine (AP) metabolism was well preserved in cryopreserved slices, but the amounts of phase I and phase II AP metabolites produced over a 3-h incubation period were markedly reduced relative to fresh slices (-58 to -71%). When cryopreserved slices were cultured for 24 h after thawing, the viability was markedly reduced, as reflected by the almost complete absence of MTT reduction and the loss of ATP content. Histological examinations showed extensive cellular necrosis. The amount of AP metabolites produced by cryopreserved slices was similar after a 3- or a 24-h culture period, indicating that AP metabolism capacities were lost at 24 h culture. In conclusion, our results suggest that cryopreserved rat liver slices may be a useful model for short-term in vitro determination of drug metabolism pathways. Further work is required to extend their use for toxicological studies.

Adenosine↗

[Computerization experience in a day hospital of integrated radiochemotherapy].

INTRODUCTION: We report on a cost-effective easy-access software developed for the functional integration of the clinical records and history of oncologic patients with the management of the Day Hospital of the Radiotherapy Department of the University Hospital A. Gemelli, in Rome. MATERIAL AND METHODS: The software was designed to archive the clinical records and history of oncologic patients and the relative chemotherapy, to manage the examination scheduling, to draw up nursing files with the planned therapy and to make statistical analyses of the department activity. Five forms are available: the patient form, recording patient data; the admission form, recording the type of therapy (e.g., chemotherapy, tests, medical examinations, etc.), the relative cost and chemotherapy protocol, detailing for instance the type of drug; the nursing file, detailing chemotherapy schedule and the dilution of each drug; the menu, to select and retrieve any record. The minimum configuration requires a 386 Intel CPU, 4 Mb RAM and 4 Mb free on the hard disk. The software is the File Maker Pro 2.1 for Windows which can interact with Apple Macintosh computers. RESULTS: Since October, 1995, we have saved the clinical records of 272 oncologic outpatients (2415 entries in all), with a mean of 201/month. This computer system permitted us to save and retrieve data for both clinical and didactic purposes and to plan our activity. CONCLUSIONS: One year after it was implemented and used in clinical practice, the system is a cost-effective and user-friendly tool for the management of the Radiochemotherapy Day Hospital of our Radiotherapy Department.

Combined Modality Therapy↗

Clinical and endocrine spectrum in patients with the 45,X/46,XY karyotype.

Cytogenetic and endocrine studies were performed in five unrelated 45,X/46,XY individuals in an attempt to correlate them with their clinical expression and gonadal morphology. A lack of a consistent pattern between cytogenetic findings and phenotype was observed. Endocrine studies revealed a wide spectrum of hypothalamic, pituitary, and gonadal hormone production as assessed by the base line levels of LH, FSH, T, and delta 4-A and their responses to appropriate exogenous stimulation (LH-RH and HCG). An adequate correlation between endocrine findings with gonadal morphology and phenotype could be established; thus demonstrating that patients with this particular chromosome complement have a functional integrity of the gonadotropin hypothalamic pituitary activity modulated accordingly with the gonadal function of each particular case.

Adolescent↗

Pattern electroretinogram, visual evoked potential and psychophysical functions in maculopathy.

To compare pattern electroretinograms and visual evoked potentials with psychophysical examinations, such as visual acuity, static (automated) perimetry and color vision in unilateral maculopathies of various origins, 20 patients with unilateral retinal diseases within the macula and the posterior pole were tested. Pattern electroretinography, visual evoked potential testing and static perimetry (Octopus program M1) were performed with three different test field sizes (20 degrees x 20 degrees, 10 degrees x 10 degrees and 6 degrees x 6 degrees). The best correlation in all three test field sizes was found between visual acuity, static perimetry and visual evoked potential. This result is surprising, since central area defined functions (visual evoked potentials, visual acuity) correlated well with a total area integrating function (mean defect in static perimetry. The pattern electroretinogram, which seems to reflect an area-related function as well, showed a correlation to static perimetry only in the smaller 10 degrees x 10 degrees and 6 degrees x 6 degrees fields and not a significant correlation in the 20 degrees x 20 degrees field. Smaller stimulation fields may therefore produce sharper results in pattern electroretinographic testing. There was no correlation between pattern electroretinograms and visual evoked potentials or visual acuity. The pattern electroretinogram was recorded under monocular and binocular viewing conditions. In 60% of the patients, the amplitude of the affected eye was more reduced in the monocular than the binocular viewing condition; the healthy fellow eye controlled stable fixation of the affected eye more readily during binocular pattern electroretinogram registration. The degree of the color vision disturbance (C-index, desaturated panel D-15 test) did not correlate to any of the other examinations.

Adolescent↗

Signaling from E-cadherins to the MAPK pathway by the recruitment and activation of epidermal growth factor receptors upon cell-cell contact formation.

E-cadherins are well characterized cell surface molecules expressed in epithelial cells, which play a major role in cell adhesion through the establishment of calcium-dependent homophilic interactions at sites of cell-cell contacts. They are also integral components of morphogenetic programs controlling the maintenance of the structural and functional integrity of epithelia. Accumulated evidence indicates that the E-cadherin-mediated cell adhesion system is highly regulated from inside the cells by a number of intracellular signaling pathways. Recently available information suggests that E-cadherins may also play a role in the transduction of signals from the outside of the cell to the cytoplasm. However, the nature of the biochemical routes regulated by E-cadherins is still largely unknown. In this study, we set out to explore the possibility that E-cadherins may regulate the activity of MAPK, a key signaling pathway involved in cell fate decisions, upon the formation of cell-cell contacts among neighboring cells. By using an immortalized non-tumorigenic keratinocyte cell line, HaCat, as a model system, we provide evidence that the assembly of calcium-dependent adherens junctions leads to a rapid and remarkable increase in the state of activation of MAPK and that this event is mediated by E-cadherins. Furthermore, we found that E-cadherins stimulate the MAPK pathway through the ligand-independent activation of epidermal growth factor receptors and the consequent activation of a biochemical route leading to the stimulation of MAPKs. These findings suggest that E-cadherins can initiate outside-in signal transducing pathways through the engagement of tyrosine kinase receptors for epidermal growth factor, thus providing a novel molecular mechanism whereby these cell adhesion molecules may ultimately control the fate of normal and transformed epithelial cells.

Cadherins↗

Contribution of the endothelin system to the renal hypoperfusion associated with experimental congestive heart failure.

The objective of this study was to define further the local activation of endothelin-1 (ET-1) and the ETA receptor as well as the functional consequences of activated ET-1 for renal hypoperfusion associated with experimental congestive heart failure (CHF). We studied eight rabbits permanently instrumented with Doppler flow probes around the renal arteries before and after the induction of epinephrine-induced CHF. CHF was characterized by left-ventricular dysfunction (fractional shortening 34+/-2% vs. 46+/-3%; p < or = 0.05) and dilatation (LVEDd 13.6+/-0.3 vs. 11.5+/-0.4 mm; p < or = 0.05), decreased mean arterial pressure (59.4+/-2.9 vs. 74.6+/-3.7 mm Hg; p < or = 0.05), increased heart rate (236+/-11 vs. 216+/-8 beats/min; p < or = 0.05) and renal vasoconstriction (vascular resistance 49.65 +/-8.55 vs. 24.61+/-5.85 U; p < 0.05; blood flow velocity, 1.58+/-0.21 vs. 3.63+/-0.31 kHz; p < 0.05). ET-1 concentrations were significantly increased not only in plasma (7.67+/-0.47 vs. 4.56 +/-0.69 pg/ml; p < 0.05) but also in renal tissue (4.8+/-0.5 vs. 3.5 +/-0.64 pg/mg; p < 0.05). Northern analysis revealed an unchanged expression of ETA receptor messenger RNA (0.79+/-0.05 vs. 0.77+/-0.04 arbitrary units; NS) in renal tissue, whereas expression of prepro-ET-1 was below the range of detection. In CHF, selective ETA-receptor antagonism with BQ-123 (1 mg/ kg bolus, i.v.) significantly increased renal blood flow velocity (3.07+/-0.38 vs. 1.33+/-0.19 kHz; p < 0.05) and reduced renal vascular resistance (29.63+/-6.22 vs. 58.17+/-8.75 U; p < 0.05) without significant effects on mean arterial pressure or heart rate. These studies demonstrate activation of the renal ET system, unaltered gene expression, and functional integrity of the renal ETA receptor in CHF. They indicate a principal functional role for the ETA receptor in renal vasoconstriction and suggest blockade of the renal ETA receptor as an important strategy to attenuate renal hypoperfusion in CHF.

Animals↗

Neural stem cells: plasticity and their transdifferentiation potential.

The presence of resident stem cells in adult tissues is of fundamental importance for the maintenance of their structural and functional integrity. In fact, throughout life, somatic stem cells attend to the critical function of substituting terminally differentiated cells lost to physiological turnover, injury or disease. Thence, one of the basic dogmata in tissue biology holds that the differentiation potential of an adult stem cell is restricted to the generation of the mature cell lineages found in the tissue to which the stem cell belongs. A plethora of recent evidences from many groups, including ours, is now providing evidence that adult stem cells may possess a broader differentiation repertoire than expected and that their fate potential may not be as tissue specific as once thought. The initial example of an unforeseen, trans-germ layer plasticity - that seems now to emerge as a prototypic functional trait of various somatic stem cells of different origin - has come from the reported awakening of a latent hemopoietic developmental capacity in stem cells isolated from the adult mammalian brain following their transplantation into sub-lethally irradiated mice. More recently, it has been shown that adult neural stem cells can differentiate into a wide array of bodily cells of different origin when injected into the blastocyst and into myogenic cells when transplanted into the adult regenerating skeletal muscle. Moreover, bone marrow stem cells can now give rise to skeletal muscle, hepatic and brain cells, whereas muscle precursors can generate blood cells. In this article, we review some of the basic notions regarding the functional properties of the adult neural stem cells and discuss findings in the expanding area of trans-germ layer conversion, with emphasis on the neural stem cell.

Animals↗

Can electrophysiological assessments of brain function be useful to the intensive care physician in daily clinical practice?

Changes in electroencephalogram parameters and auditory event-related potentials, induced by interruption to propofol sedation in intensive care patients, provide a number of electrophysiological measures that can be used to assess neurological function accurately. Studies of electroencephalogram parameters suggest that power spectral estimation, as root mean square power, is more useful and precise than spectral edge frequency 95% in evaluating the functional integrity of the brain. When such parameters are used to evaluate neurological function, in particular the N100 and mismatch negativity components, a precise assessment of a patient's readiness to awaken from a pharmacologically induced coma (such as sedation) can be obtained. In terms of ease of use, however, it is more difficult to establish whether N100 or mismatch negativity is superior.

Electroencephalography↗