[FUNCTION AND ORGANIZATION OF MEDICAL VERIFICATION. (REPORT OF THE SOCIAL SECURITY MEDICAL HIGH COMMISSION)].
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The role of serotonin (5HT2) receptor blockade in myocardial reperfusion injury: effects of LY53857 in a canine model of myocardial infarction.
The potential protective effects of serotonin receptor antagonism during the process of acute myocardial infarction were studied in anesthetized male dogs, which were subjected to a 90-min left circumflex coronary artery occlusion followed by 5 h of reperfusion. Either vehicle (0.9% NaCl) or the serotonin (5HT2) receptor antagonist LY53857 was infused i.v. at a dose of 0.5 mg/kg, followed by a constant infusion of 2 mg/kg/min beginning 5 min before left circumflex coronary artery occlusion and continuing throughout the duration of the ischemia and subsequent reperfusion. Verification of functional 5HT2 receptor antagonism in the circulating blood of the LY53857-treated dogs was monitored throughout the experiments by periodic assessment of ex vivo platelet reactivity to exogenous serotonin. After 5 h of reperfusion, the hearts were excised and analyzed utilizing histochemical staining with triphenyltetrazolium, which demarcates myocardial infarct size and anatomical area of myocardium at risk of infarction. There was not a significant reduction of infarct size with LY53857 treatment: control infarct/area at risk = 38.6 +/- 4.7%, n = 9 LY53857 infarct/area at risk = 33.4 +/- 3.8%, n = 6. Similarly, when myocardial infarct size was analyzed as a function of myocardial collateral blood flow, there were no significant effects of drug treatment on the relationship between collateral blood flow and infarct size. The effects of 5HT on neutrophil activation were determined by measuring the potential ability of 5HT to enhance the chemotactic peptide-induced production of superoxide. 5HT did not activate human neutrophils in vitro and LY53857 had no effect on neutrophil superoxide production.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
[Use of streptokinase and urokinase in deep venous thrombosis and pulmonary embolism: indications and clinical experience].
The efficacy of fibrinolysis in DVT is dependent upon the age and organization of the thrombus as well as its localization. In consequence, selective evaluation prior to determining the indication for thrombolytic therapy is just as important for therapeutic success as choosing the appropriate fibrinolytic agent. To improve the results of fibrinolysis a team of angiologists, hemostaseologists, radiologists and surgeons are cooperating in a special "thromboembolic care unit". Phlebographic criteria were defined which allow differentiation of fibrinolytic indications depending upon the site of the thrombus. Selection of the fibrinolytic agent and careful monitoring of the thrombolysis should ensure a maximum therapeutic effect with a minimum of bleeding complications. The duration of thrombolysis was established through phlebographic verification and functional tests with venous occlusion plethysmography. The cause and results of 108 cases of thrombolysis shall be presented.
Humans↗
Uneven regional distribution of nucleotide metabolism in human brain.
Adenine and uridine nucleotides and adenosine are proposed to act as neuromodulators and other nucleotides and nucleosides are also suggested to be involved in brain function. A following major step towards the verification of the functional role of nucleotides and nucleosides in the brain would be the examination of regional distribution of purines, pyrimidines and the enzymes involved in their metabolism. Using our recently developed chromatography-based assay for nucleosides from tissue homogenates, we analysed nucleosides in microdissected samples derived from various regions of human brain. Marked differences in the levels of nucleosides were measured in the cerebral cortex, cerebellar cortex, thalamus and white matter. The greatest levels of most nucleosides were found in the cerebral cortex, followed by the cerebellar cortex and the white matter while the smallest concentrations were found in the thalamus, although adenosine and xanthine showed a different distribution pattern in these brain areas. Within the cerebral cortex, the measured substances showed little variations except certain high levels in the cingulate and low levels in the frontal cortex. Even distribution of nucleosides was found in the thalamic nuclei while relative high values were measured in the medial geniculate body. Since a dramatic change in nucleoside concentrations occurs after death, the measured nucleoside concentrations are an interplay of original nucleotide and nucleoside concentrations and enzyme reactions following death. Thus our results suggest regional differences in nucleotide and nucleoside composition and nucleotide metabolising enzyme activities between brain areas.
Adenosine↗
["Mixed" changes in spirometry--verification of the pattern of lung function impairment].
UNLABELLED: Reduction of FEV1%FVC ratio together with lowered FVC often is interpreted as "mixed" ventilatory disturbances. The aim of this study was to evaluate the real frequency of the overlapping obstructive and restrictive impairments in patients with FEV1%FVC<0.7 and FVC<80% of predicted, which is often described as "mixed" ventilatory defect. MATERIAL AND METHODS: Retrospective, cross-sectional analysis of pulmonary function data. Only 748 out of 6673 patient's data met inclusion criteria: performed bodypletysmography and FEV1%FVC<0.7 together with FVC<80%pred. in spirometry. In all cases spirometry and whole body pletysmography were performed according to ERS standards by experienced staff using MasterLab-"Jaeger" equipment. Reference values according to ERS guidelines were applied. RESULTS: We found volume restriction defined as TLC below lower limit of normal in only 17% of the patients. There was also inverse correlation between FEV1%pred. and RV%TLC%pred. ratio. So, the basic mechanism of reduction in VC and FVC in patients with normal TLC is increased RV. CONCLUSION: "Mixed" ventilatory impairment in majority cases represents airway obstruction with lung hyperinflation. Such a coincidence of reduced FVC and FEV1%FVC ratio requires further investigations (pletysmography) to clarify the reason for the diminished vital capacity.
Aged↗
Diversity and versatility of lipid-protein interactions revealed by molecular genetic approaches.
The diversity in structures and physical properties of lipids provides a wide variety of possible interactions with proteins that affect their assembly, organization, and function either at the surface of or within membranes. Because lipids have no catalytic activity, it has been challenging to define many of their precise functions in vivo in molecular terms. Those processes responsive to lipids are attuned to the native lipid environment for optimal function, but evidence that lipids with similar properties or even detergents can sometimes partially replace the natural lipid environment has led to uncertainty as to the requirement for specific lipids. The development of strains of microorganisms in which membrane lipid composition can be genetically manipulated in viable cells has provided a set of reagents to probe lipid functions. These mutants have uncovered previously unrecognized roles for lipids and provided in vivo verification for putative functions described in vitro. In this review, we summarize how these reagent strains have provided new insight into the function of lipids. The role of specific lipids in membrane protein folding and topological organization is reviewed. The evidence is summarized for the involvement of anionic lipid-enriched domains in the organization of amphitropic proteins on the membrane surface into molecular machines involved in DNA replication and cell division.
Bacterial Proteins↗
The neural bases of strategy and skill in sentence-picture verification.
This experiment used functional Magnetic Resonance Imaging to examine the relation between individual differences in cognitive skill and the amount of cortical activation engendered by two strategies (linguistic vs. visual-spatial) in a sentence-picture verification task. The verbal strategy produced more activation in language-related cortical regions (e.g., Broca's area), whereas the visual-spatial strategy produced more activation in regions that have been implicated in visual-spatial reasoning (e.g., parietal cortex). These relations were also modulated by individual differences in cognitive skill: Individuals with better verbal skills (as measured by the reading span test) had less activation in Broca's area when they used the verbal strategy. Similarly, individuals with better visual-spatial skills (as measured by the Vandenberg, 1971, mental rotation test) had less activation in the left parietal cortex when they used the visual-spatial strategy. These results indicate that language and visual-spatial processing are supported by partially separable networks of cortical regions and suggests one basis for strategy selection: the minimization of cognitive workload.
Adolescent↗
Purification of functional T lymphocytes from splenocytes of the beluga whales (Delphinapterus leucas).
In an effort to gain knowledge on immune functions in beluga whales, Delphinapterus leucas, we have used two physical methods for the purification of T lymphocytes of spleen cells. Isolation by sheep red blood cells (SRBC) rosetting and by adherence on nylon wool columns were tested. SRBC-rosetting gave unreliable results in obtaining purified T cells. Therefore, the purification of T cells was done using nylon wool columns. Less than 3% of the IgM(+) B cells remained in effluent populations. In the later population, 45% gave positive staining with mouse anti-human CD4 allowing us to verify functionality of the cells. The study of calcium mobilization and tyrosine kinase activation, mediated by CD4 cross-linking permitted verification of the functionality of cells. We also showed that upon activation with mitogens, beluga T cells upregulate the density of MHC class II molecules on their surfaces. CD4 cross-linking with a specific antibody inhibited the proliferation response. Overall, the activation of beluga whales lymphocytes did not differ markedly from what is known in other species. This study can help in the groundwork for functional investigation of the beluga whale's immune system.
Animals↗
Database-assisted promoter analysis.
The analysis of regulatory sequences is greatly facilitated by database-assisted bioinformatic approaches. The TRANSFAC database contains information on transcription factors and their origins, functional properties and sequence-specific binding activities. Software tools enable us to screen the database with a given DNA sequence for interacting transcription factors. If a regulatory function is already attributed to this sequence then the database-assisted identification of binding sites for proteins or protein classes and subsequent experimental verification might establish functionally relevant sites within this sequence. The binding transcription factors and interacting factors might already be present in the database.
Binding Sites↗
Long-term monolayer culture of human anterior pituitary cells: functional validation of a method.
This study demonstrates the response of human pituitary cell monolayers to a wide variety of hormonal stimuli. Appropriate release of luteinizing hormone (LH) and prolactin (PRL) was used as a verification of cell function. Cells that had been in culture for 20 days, with no hormonal additions, were exposed to LH-releasing hormone (LH-RH) continuously for 14 days. This resulted in an immediate fivefold increase in secretion of LH followed by a depression in LH production over the remaining 10-day period. After an 8-day period without hormonal additions, the same cultures again demonstrated a threefold increase in response to retreatment with LH-RH. In two similar studies, cells that had been in culture for 28 and 31 days were treated with bromocriptine, pergolide, dopamine, or thyrotropin-releasing factor (TRF). TRF elicited an increase of PRL in the medium by nearly double the control values. The addition of dopamine, pergolide, or bromocriptine resulted in a depression of PRL during the treatment period. This study has shown that human pituitary cells maintained in long-term monolayer culture respond predictably to a wide range of hormonal stimuli.
Bromocriptine↗
Awake craniotomy in glioma surgery.
Awake craniotomy is the cheapest and most reliable method to ensure neurological integrity in cerebral gliomas that infiltrate or come close to the eloquent areas of the brain, allowing (a) the localization of eloquent cortical areas by electrical stimulation and epileptic foci by cortical recording, and (b) the monitoring of the functional integrity of awake patients while aiming at subtotal removal of the gliomatous tissue. In addition, awake craniotomy opens a brief but unique window to the living brain for (a) basic neuroscience, including verification of preoperative functional imaging data and recording of electrophysiological correlates of mental processes, and for (b) applied research, including development of innovative instrumentation for brain recording and monitoring as well as screening for potential areas to be modulated in movement disorders and chronic pain.
Brain Neoplasms↗
[New species and sympatric relations of the chigger mite species group Talmiensis (Trombiculidae, Neotrombicula)].
A revision of chigger mites species being closely related to Neotrombicula talmiensis (Schluger, 1955) has been performed. 2 new species are described: N. pontica sp. n. from Krasnodar Territory (Western Caucasus) and N. sympatrica sp. n. from Krasnodar Territory, Daghestan, Tuva, Armenia, Kirghizia and Turkey (Rize Province). N. pontica sp. n. is closely related to N. carpathica Schluger et Vysotzkaya, 1970 and differs from this species by the larger number of idiosomal setae (NDV = = 75-99 against 63-77), shorter legs (Ip = 782-847 against 844-920, TaIII = 67-74 against 70-80), lesser m-t (0.180 against 0.192), slightly lesser scutum and slightly longer setae. N. sympatrica sp. n. is closely related to N. carpathica and differs from this species by the longer scutal and idiosomal setae (PL = 67-78 against 57-69, H = 65-75 against 57-68, Dmin = 43-52 against 39-48, Dmax = 61-70 against 54-64), longer legs (Ip = 892-973 against 844-920, TaIII = = 77-86 against 70-80) and lesser m-t (0.168 against 0.192). N. carpathica is reported for the first time from Northern Caucasus (Karachai-Cherkess Republic, Kabardino-Balkaria, North Ossetia); N. talmiensis is reported for the first time from Khakasia. Data on joint occurrence of 3 species are reported. The 3 types of sympatric pairs of the species have been found in the Western Caucasus: 1) N. pontica sp. n. and N. sympatrica sp. n., 2) N. sympatrica sp. n., and N. carpathica, 3) N. pontica sp. n., and N. carpathica. Differences between species in that localities, where the joint occurrence was recorded, have been analysed. Sympatric relations between N. sympatrica sp. n. and N. carpathica are characterized by increased value of some general diagnostic characters of these species, such as the length of idiosomal setae and length of legs. Different characters play main distinguishing role in different sympatric localities. Besides that, some local features appear in certain species of sympatric pairs (narrow scutum in 3 samples of N. carpathica, numerous idiosomal setae in 1 sample of N. sympatrica), that differs the populations of the sympatric pair, and also discriminates such aberrant group from other material of the same species. Functions produced by the method of discriminant analysis are proposed for the purpose of diagnostics in talmiensis group. Tests for assumptions of discriminant analysis, such as normality and homogeneity of variances, has been performed beforehand. Verifications of the functions on test samples demonstrated the good quality of its working. Key to species using discriminant functions has been constructed.
Animals↗
[Experimental verification and experience in the use of functional pneumoepidurography in the diagnosis of lumbar osteochondrosis].
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Adolescent↗
The Helicobacter pylori genome: from sequence analysis to structural and functional predictions.
Fold assignments for proteins from the Helicobacter pylori genome are carried out using BASIC, a profile-profile alignment algorithm recently tested on the Mycoplasma genitalium and Escherichia coli genomes. The fold assignments are followed by automated function evaluation, based on the multilevel description of functional sites in proteins. Over 40% of the proteins encoded in the H. pylori genome can be recognized as belonging to a protein family with known structure. Previous estimates suggested that only 10-15% of genome proteins could be characterized this way. This dramatic increase in the number of recognized homologies between H. pylori proteins and structurally characterized protein families is partly due to the rapid increase of the database of known protein structures, but mostly it is due to the significant improvement in prediction algorithms. Knowledge of a protein fold adds a new dimension to our understanding of its function and, similarly, structure prediction can also add to understanding, verification, and/or prediction of function for uncharacterized proteins. Several examples analyzed in more detail in this article illustrate insights that can be achieved from structure and detailed function prediction.
Algorithms↗
Use of bar codes in inpatient drug distribution.
The development and operation of a prototype inpatient drug distribution system that uses bar codes is described, and the impact of bar coding on the cassette-filling and verification process is summarized. A prototype pharmacy dispensing site was created to function in parallel with an existing satellite dispensing site that served 78 general medical-care beds. Supplemental labels encoded with an 11-digit unique product identification number, a 5-digit expiration date, and a 6-character lot number were generated and affixed to all unit dose packages dispensed from the prototype pharmacy site. The unit doses were labeled with Code 49 symbology; each label measured 0.8 x 1.25 inches. Each patient cassette was labeled using Code 39 symbology. A cost-benefit model was developed, and the two dispensing systems were compared with respect to (1) time to fill patient cassettes, (2) time to verify patient cassettes, (3) time to process patient charges and credits, (4) time to correct dispensing errors, (5) accuracy of the cassette-filling process, and (6) accuracy of the cassette verification process. Bar-code dispensing and verification saved 1.52 seconds per dose. Additionally, the cassette verification function was shifted from pharmacists to technicians. Estimated per-dose cost of the bar-code system was 2.73 cents. A measurable improvement in the accuracy of filling patient cassettes was documented. The feasibility of using bar codes in unit dose dispensing was demonstrated, and the prototype system was shown to produce cost efficiencies and patient-care benefits.
Clinical Pharmacy Information Systems↗
Chromatographic investigation of the glycosylation pattern of alpha-1-acid glycoprotein secreted by the HepG2 cell line; a putative model for inflammation?
In certain pathophysiological conditions, such as rheumatoid arthritis, there are alterations in the glycosylation pattern of the acute phase protein, alpha-1-acid glycoprotein (AGP). These changes are likely to be functionally significant, however, verification of the latter role requires a system which reflects in vivo glycosylation changes in AGP and also produces sufficient quantities of the protein for further study. The human hepatoma cell line HepG2 is documented as displaying a shift in the glycosylation pattern of glycoproteins from normal state to acute phase after stimulation with inflammatory mediators. We have isolated AGP from the culture medium of HepG2 cells both before and after stimulation with a cytokine preparation and analysed the glycosylation pattern of each preparation, after enzymatic release, by high pH anion-exchange chromatography. Before stimulation, the glycosylated population was similar to a profile of AGP isolated from normal plasma; however, cytokine stimulation resulted in a shift to a profile which was consistent with that of AGP from a rheumatoid arthritis sufferer. Thus a HepG2 cell culture system is capable of being a crude model of the changes in glycosylation of acute phase proteins although it has a tendency to produce oligosaccharide chains which are not fully sialylated.
Animals↗
Human phenylalanine hydroxylase gene expression in kidney and other nonhepatic tissues.
Phenylalanine hydroxylase (PAH) is the key enzyme in phenylalanine metabolism. PAH deficiency results in hyperphenylalaninemia, leading to severe mental retardation in the classical form of the disease, phenylketonuria (PKU). Previously the expression of PAH could only unambiguously be demonstrated in human liver, whereas in rodents PAH expression has been established in kidney and liver. Reports concerning PAH activity in other human or rodent tissues were severely questioned by subsequent investigations such that they did not gain general recognition. Conducting Northern blot analyses, we detected the PAH transcript in RNA isolated from human liver, kidney, pancreas, and brain. PAH gene expression in human kidney was subsequently investigated by RNase protection assay analyses, RNA in situ hybridization, immunohistochemistry, enzyme assay, and cDNA isolation. These experiments allowed the conclusive verification of a functional PAH enzyme in human kidney. The primary structure of the kidney transcript corresponded to the structure of the liver transcript. Human kidney PAH may play a significant role in phenylalanine homeostasis of the organism, as impaired phenylalanine hydroxylation has been observed in renal failure and differences in the regulation of the kidney versus the liver enzyme have been indicated. These results provide new aspects to research into the basis for the heterogeneity of hyperphenylalaninemia phenotypes and establish that the expression of the human PAH gene is not limited to the liver.
Blotting, Northern↗
A comparison of neural circuits underlying auditory and visual object categorization.
Knowledge about environmental objects derives from representations of multiple object features both within and across sensory modalities. While our understanding of the neural basis for visual object representation in the human and nonhuman primate brain is well advanced, a similar understanding of auditory objects is in its infancy. We used a name verification task and functional magnetic resonance imaging (fMRI) to characterize the neural circuits that are activated as human subjects match visually presented words with either simultaneously presented pictures or environmental sounds. The difficulty of the matching judgment was manipulated by varying the level of semantic detail at which the words and objects were compared. We found that blood oxygen level dependent (BOLD) signal was modulated in ventral and dorsal regions of the inferior frontal gyrus of both hemispheres during auditory and visual object categorization, potentially implicating these areas as sites for integrating polymodal object representations with concepts in semantic memory. As expected, BOLD signal increases in the fusiform gyrus varied with the semantic level of object categorization, though this effect was weak and restricted to the left hemisphere in the case of auditory objects.
Adult↗