Inter-rater reliability of postural observation after stroke.

Objective: To explore the inter-observer reliability of bedside observations of stroke patients' posture using two versions of a pictorial tool.

Design: Three projects were conducted. The initial version of the tool was used in project 1.

Heated dorsal hand vein sampling for metabolic studies: a reappraisal.

The purpose of the current study was to determine the accuracy of a given heated dorsal hand vein (HDHV) measurement in predicting a simultaneous arterial measurement and to validate this technique for use in stable isotope studies. Twenty catheterizations of the femoral artery, femoral vein, and a dorsal hand vein were performed in 13 healthy male subjects. Simultaneous blood samples were obtained from all three sites during primed continuous infusions of L-[1-13C]leucine (Leu) and L-[ring-2H5]phenylalanine (Phe) in the postabsorptive state, with or without intravenous glucose infusion.

Positioning of the stroke patient: a review of the literature.

Stroke is a common and disabling illness, adversely affecting the quality of life of hundreds of people each year. While there are many therapeutic approaches to stroke patient rehabilitation, encouraging patients to adopt "reflex-inhibiting" patterns of posture is a widely advocated strategy for helping patients to avoid complications of hemiplegia such as spasticity and contractures. However, while the central role of nurses in thus helping patients is recognized, the influence of posture on recovery from stroke has never been evaluated.

Insulin increases transcription of rat gene 33 through cis-acting elements in 5'-flanking DNA.

Gene 33 is a multihormonally-regulated rat gene whose transcription is rapidly and markedly enhanced by insulin in liver and cultured hepatoma cells. To examine the mechanism by which insulin regulates transcription, we have constructed chimeric plasmids in which expression of the bacterial cat gene, encoding chloramphenicol acetyltransferase (CAT), is governed by gene 33 promoter elements and contiguous sequences in DNA flanking the transcription start point (tsp). When transfected into H4IIE hepatoma cells, these constructs gave rise to stably transformed cell lines producing the bacterial CAT enzyme.

State-dependent fear extinction with two benzodiazepine tranquilizers.

Four experiments with rats were run to investigate whether fear extinction conducted under the influence of a benzodiazepine transfers to the undrugged state. Fear was conditioned by pairing an experimental chamber with footshock and was assessed by observing freezing, a characteristic response of the rat to stimuli associated with shock. In Experiment 1, extinction of the chamber cues under chlordiazepoxide (librium) or diazepam (valium) was compared with extinction under a placebo; both drugs interfered with extinction in a dose-dependent manner as indicated by freezing during an undrugged test.

Molecular cloning and analysis of full-length cDNAs cognate to a rat gene under multihormonal control.

Gene 33 is a multihormonally regulated rat gene whose transcription is rapidly and markedly enhanced by glucocorticoids, insulin, or cAMP. A cDNA clone (p216) containing a nearly full-length DNA complementary to the mRNA of this gene was isolated from a cDNA library and sequenced. The cDNA represents the entire mRNA transcript, except for three bases at the 5' terminus.

Structure of a multihormonally regulated rat gene.

Gene 33, a rat gene transcriptionally enhanced by glucocorticoids, insulin, or cyclic AMP, was isolated from a library of rat genomic DNA and characterized by sequence comparison to a full-length cDNA. The structural gene spans 13,500 bp encoding 2970 bp of exon sequences interrupted by three introns of about 9600, 101 and 811 bp, respectively. Exons (5' to 3') are 198, 194, 77 and 2501 bp in length; the first of these initiates at the transcriptional start point determined by S1 nuclease mapping.

Gene-specific acquisition of hormonal responsiveness in rat liver during development.

Cloned cDNAs were used in hybridization analyses to assess hormonal responsiveness of two similarly regulated genes in livers of late-term fetal rats. Transcription of the tyrosine aminotransferase gene and of gene 33 (Lee et al.: J Biol Chem 260:16433-16438, 1985) is enhanced by glucocorticoids and by each of the usually antagonistic hormonal agents, insulin and cAMP, in adult liver, and that of both genes is developmentally activated at or just prior to birth.

Changes in hepatic differentiation following treatment of rat fetuses with 5-azacytidine.

Rat fetuses of 20 days gestational age were treated in utero with 5-azacytidine. Within 14 to 18 h after treatment several significant changes in the fetal livers were observed, including a dramatic maturation of hepatocyte morphology with little alteration in hematopoietic elements. Assessment of mRNA levels by hybridization to cloned cDNAs, together with other measures of gene expression, established that the change in hepatocyte morphology was associated with strong activation of expression of genes normally activated later in development, including those coding for the liver enzymes tyrosine aminotransferase and phosphoenolcarboxykinase and a gene of unknown specificity that is regulated in liver much like the aminotransferase.

Different mechanisms control developmental activation of transcription of genes subject to identical hormonal regulation in adult liver.

Developmental changes in expression of two genes subject to identical hormonal controls in adult liver were examined in livers of fetal and newborn rats. Both mRNA concentrations and transcription of tyrosine aminotransferase were very low throughout gestation and increased sharply at birth. The mRNA of gene 33 (Lee et al.

Insulin enhances transcription of the tyrosine aminotransferase gene in rat liver.

The mechanism of insulin-mediated induction of tyrosine aminotransferase in rat liver was investigated using a cloned cDNA probe. The level of aminotransferase mRNA increases about fourfold following administration of the hormone. This induced mRNA accumulation does not require de novo protein synthesis.

Molecular cloning of cDNAs cognate to genes sensitive to hormonal control in rat liver.

Poly(A)-RNAs were prepared from livers of rats treated with hydrocortisone and cycloheximide, then enriched for large mRNAs by successive sucrose gradients and gel electrophoresis. The size-selected RNAs were used as templates for synthesis of double-stranded cDNAs that were cloned in Escherichia coli using the pBR322 plasmid vector. Recombinant plasmids characterized as carrying inserts of potential interest were further analyzed by differential hybridization to mRNAs from untreated and hydrocortisone-treated rats.

Activation of tyrosine aminotransferase expression in fetal liver by 5-azacytidine.

Rat fetuses of 20 days gestational age were treated in utero with the inhibitor of DNA methylation, 5-azacytidine. The liver enzyme tyrosine aminotransferase, normally expressed at very low levels until several hours after birth, was increased by the drug in the fetal livers after a lag period of about 9 hours, reaching a level 70-fold above control levels 18 hours after treatment. The high levels attained after 5-azacytidine treatment are comparable to those of glucocorticoid-treated adult livers, and were not further increased by administration of hydrocortisone to dams carrying treated fetuses.

Development of tyrosine aminotransferase in perinatal rat liver: changes in functional messenger RNA and the role of inducing hormones.

Expression of the hepatic enzyme tyrosine aminotransferase was analyzed in the perinatal period of development in the rat, when this expression undergoes significant changes associated with hepatocyte differentiation. In late prenatal liver both enzyme and functional mRNA gene products are present at levels 10- to 15-fold below those in the fully differentiated adult liver. This low level of expression in fetal liver is refractory to induction by glucocorticoids, but both gene products are increased to a limited extent by cyclic AMP.

Concanavalin A alters the turnover rate of tyrosine aminotransferase in cultured hepatoma cells.

Concanavalin A added to monolayer cultures of Reuber H-35 hepatoma cells caused a rapid inactivation of tyrosine aminotransferase (L-tyrosine:2-oxoglutarate aminotransferase, E.C. 2.

Selective effect of the metallocarcinogen beryllium on hormonal regulation of gene expression in cultured cells.

Effects of the metallocarcinogen beryllium on regulation of gene expression were assessed by analysis of hormonal regulation of synthesis of tyrosine aminotransferase in beryllium-treated hepatoma cell cultures. Cell growth was not affected by exposure of the cells to 1 micro M BeSO4 throughout their 4- to 5-day growth cycle. In cells pretreated in this way, the induction by glucocorticoids was specifically impaired, the extent of induced enzyme synthesis being reduced about 50%.

Effects of insulin on messenger RNA activities in rat liver.

Liver poly(A) RNA, isolated from adrenalectomized rats after insulin treatment, was translated in a nuclease-treated lysate of rabbit reticulocytes and quantitated for both total activity and the capacity to synthesize the insulin-inducible enzyme tyrosine aminotransferase. Analysis of the translated products from poly(A) RNA isolated 1 h after insulin treatment showed a 2.7-fold increase in activity of tyrosine aminotransferase mRNA.

Changes in hepatic levels of tyrosine aminotransferase messenger RNA during induction by hydrocortisone.

Messenger RNA specific for tyrosine aminotransferase was quantitated by microinjection into oocytes of Xenopus laevis. The heterologously translated enzyme was identified by specific immunoprecipitation and found to be identical with authentic aminotransferase by several criteria. The level of functional message present in rat liver increases during hydrocortisone induction, and this increase is directly proportional to the increased rate of synthesis of the enzyme.

Role of coenzyme in aminotransferase turnover.

The role of coenzyme in determining intracellular contnet of pyridoxal enzymes was assessed by analyzing effects of pyridoxine deficiency on the rapidly degraded, readily dissociable tyrosine aminotransferase (EC 2.6.1.

Separation of cellular and viral DNA polymerases by affinity chromatography on polynucleotide-Sepharose.

Polyguanylate- and poly(2'-O-methyl)uridylate-Sepharose have been prepared for affinity chromatography of DNA polymerases of viral origin (reverse transcriptase). Both cellular DNA polymerases and reverse transcriptase bind to polyguanylate-Sepharose. The cellular polymerases can be eluted from the column between 0.

Differential degradation of messenger RNAs in mammalian cells.

Through the use of an assay that measures cellular capacity for specific enzyme synthesis, mRNA of alanine aminotransferase (EC 2.6.1.