Search PubMed⌕ Search

Biomedical subjects

J Rose

Publications and source records attributed to J Rose.

At least 127 records · Page 7Linked to original sources

The thumb's knuckle. Flexibility in the thumb subdomain of T7 RNA polymerase is revealed by the structure of a chimeric T7/T3 RNA polymerase.

We have solved the structure of a chimeric T7/T3 RNA polymerase (RNAP) in an orthorhombic crystal by molecular replacement with the T7 RNAP structure determined from a monoclinic crystal. The structure of the protruding "thumb" subdomain of the polymerase appears very different in these two crystals apparently because of differences in packing contacts made by the thumb subdomain. These observations support the proposal that the thumb subdomain is flexible and can wrap around bound template to obstruct polymerase: template dissociation during processive synthesis.

Amino Acid Sequence↗

Crystals of glutamine-binding protein in various conformational states.

Crystals of glutamine-binding protein (GlnBP) in various conformational states have been obtained. Crystals of the ligand-free "open" state (denoted form B) have unit cell dimensions a = 86.3 A, b = 86.3 A, c = 81.5 A, alpha = beta = gamma = 90 degrees and diffract to about 2.3 A resolution. An analysis of the intensity data using an Requiv plot indicates that the crystal system is orthorhombic, space group P2(1)2(1)2(1). Crystals of the ligand-bound "open" state (form B*) are obtained by soaking form B crystals with glutamine (Gln) and diffract to about 1.9 A. Crystals of the GlnBP-Gln complex in a ligand-bound "closed" state (form C) belong to space group P2(1)2(1)2(1) with a = 62.0 A, b = 65.7 A and c = 121.8 A and diffract to about 2.3 A. Crystals of a selenomethionyl GlnBP (form B') are isomorphous to form B crystals and diffract to about 2.1 A resolution.

Carrier Proteins↗

New crystal forms of a micro-class glutathione S-transferase from rat liver.

Two new crystal forms of isoenzyme 3-3 of rat liver glutathione S-transferase (GST 3-3) have been obtained. They were grown under essentially the same crystallization conditions as those reported for the C2 crystal form [Fu, Rose, Chung, Tam & Wang (1991). Acta Cryst. B47, 813-814]. The new crystals belong to space group P2(1) with one form having cell dimensions a = 101.6, b = 69.5, c = 81.4 A, and beta = 113.6 degrees, and the other form having cell parameters a = 97.4, b = 81.1, c = 69.4 A and beta = 109.2 degrees. These new crystals diffract to at least 2.5 A, resolution. The molecular packing arrangements in these P2(1) crystals have been found by molecular replacement studies.

Journal Article↗

The octameric histone core of the nucleosome. Structural issues resolved.

The crystal structure of the histone octamer has now been determined at 3.1 A resolution and refined to a crystallographic R value of 25.5%. The overall shape of the structure is significantly different from that originally reported by Burlingame et al. and its length is now in agreement with that observed by Klug et al. in their low-resolution studies. The experimental intensity data used in constructing the new electron density map were the same as those used by Burlingame et al. for the original electron density map. In addition, the methods used in producing the new density map were also the same as those for the original map. The only difference between the two calculations was the selection of the heavy-atom location. The large change seen in the structural image (110 A x 70 A x 70 A versus 55 A x 70 A x 70 A) was due to a relatively small change (2.27 A shift) of the heavy-atom site. The fact that the shape and size of the original structure were incorrect is surprising and unusual, since the electron density map that produced the original model was clear for most parts of the structure; one could easily see the well-formed right-handed helices of the H2A and H2B molecules, and the ordered parts of the H2A and H2B molecules could be easily traced from end to end. A comparison of the two maps shows that the original image was derived from two fused copies of the correct structure rotated by +/- 120 degrees from its true location along a rotation axis parallel to the z-axis and the image seen was a partial (about 19.5%) overlap of two molecules. An explanation is given as to how such a small shift of the heavy-atom position could create such a double image in the unit cell, and how the original electron density map could be converted to the new map by a phase modification in the Fourier synthesis. This study resolves the differences between the analyses of the shape and size of the histone octamer structure.

Crystallography, X-Ray↗

A placebo-controlled evaluation of single, escalating doses of CL 284,846, a non-benzodiazepine hypnotic.

This report describes the first evaluation in humans of CL 284,846, a non-benzodiazepine compound with a preclinical profile indicative of sedative/hypnotic properties. Healthy, normal male volunteers were assigned randomly to receive single oral doses of 1, 5, 15, 30, or 60 mg of CL 284,846 or placebo on a double-blind basis. Observations were made over the subsequent 25 hours to determine the safety, pharmacokinetic profile, and psychometric effects of the test compound. CL 284,846 was well tolerated in the normal volunteers, causing no significant changes in vital signs, EEG, ECG, hematologic, or clinical chemistry laboratory parameters. Although few adverse events were noted at doses less than 60 mg, at the highest dose (60 mg), all volunteers reported transient neurologically related adverse events (e.g., impaired concentration, difficulty focusing, and impaired coordination), reflecting the central nervous system action of the compound. Although determination of hypnotic efficacy was not an objective in this Phase I study, daytime treatment with 60 mg of CL 284,846 was associated with greater reports of drowsiness and impaired performance on psychomotor tests. However, memory, as assessed by a word recall test, was not affected at any dose of the compound. Pharmacokinetic analyses revealed CL 284,846 to be absorbed and eliminated rapidly (Tmax = 0.9-1.5 hr, T 1/2 = 0.9-1.1 hr), with a dose-proportional AUC (area under cure). Plasma levels of CL 284,859, the primary desethylated metabolite of CL 284,846, were much lower in humans than in other species, indicating that the metabolism of CL 284,846 in humans may differ from that of rodents and dogs. Overall, CL 284,846 was well tolerated, and the authors recommend repeating dose safety and pharmacokinetic studies in healthy volunteers.

Acetamides↗

Muscle pathology and clinical measures of disability in children with cerebral palsy.

We performed a histologic and morphometric study of spastic muscle from 10 children with diplegic cerebral palsy, comparing muscle structure with the gait parameters of energy expenditure index and dynamic electromyography. Variations in fiber area within and between fiber types were increased significantly in children with cerebral palsy. In each of the control subjects, the combined coefficient of variation for type-1 and type-2 fiber area was less than 25% and the average was 17%; in the subjects with cerebral palsy, the combined coefficient of variation was more than 25% and the average was 36% (p < or = 0.004). The average difference between the mean area of type-1 and type-2 fibers was 26.7 +/- 18.9% for subjects with cerebral palsy and 4.2 +/- 2.4% for control subjects (p < or = 0.004). There was a 67% predominance of one fiber type in the subjects with cerebral palsy compared with a 55% predominance in the control subjects (p < or = 0.03). The difference between the total area of type-1 and type-2 fibers was 57% in the subjects with cerebral palsy and 17% in the control subjects (p < or = 0.002). There was a significant correlation between the combined coefficient of variation of fiber area and the energy expenditure index (r = 0.77, p < or = 0.03). The difference between the mean area of type-1 and type-2 fibers correlated with prolongation of electromyographic activity (r = 0.69, p < or = 0.05). No abnormalities in fiber ultrastructure were found in the subjects with cerebral palsy. Children with cerebral palsy had abnormal variation in the size of muscle fibers and altered distribution of fiber types. The values for variation in fiber area correlated with the energy expenditure index and with prolongation of electromyographic activity during walking.

Adolescent↗

Passive immunization of cynomolgus macaques with immune sera or a pool of neutralizing monoclonal antibodies failed to protect against challenge with SIVmac251.

In the first of two passive transfer experiments, three groups of four macaques were injected intraperitoneally with a normal serum pool, an immune serum pool (pool 1) collected 132-172 weeks postinfection with the 11/88 pool of SIVmac251, or with a pool of four neutralizing monoclonal antibodies (KK9, 17, 54, and 56) raised against gp120 of the 11/88 pool. Sera were given at a dose of 13 ml/kg whereas the MAb pool was given at 30 ml/kg. In a second experiment, a further four macaques were injected with an immune serum pool (pool 2) collected 12 weeks postinfection with simian-grown SIVmac251 at a dose of 19 ml/kg. Animals in both experiments were challenged with SIVmac251 grown in simian peripheral blood lymphocytes. Despite high levels of circulating antibodies in the serum of animals that received either the immune serum pools or the MAbs, all macaques became infected following challenge. The results described are in contrast to a previous report in which passive transfer of sera from animals infected with SIVsm successfully protected against challenge with the homologous virus grown in human PBMCs. Challenge with SIVmac251 grown in simian PBMCs may be the reason for these conflicting results. Nevertheless, the results suggest that in this model the presence of circulating neutralizing antibodies alone does not necessarily confer protection against challenge with SIVmac251 grown in simian cells.

Animals↗

Involvement of activation of ATP-dependent potassium channels in ischemic preconditioning in swine.

This study evaluated the importance of ATP-dependent potassium channels (KATP) for ischemic preconditioning (IP) in swine. Swine were studied because due to the sparsity of their innate collateral circulation infarct size (IS) development closely resembles that observed in humans. Ninety minutes of ischemia at a blood flow reduction sufficient to reduce regional myocardial work by 90% caused 13.2 +/- 8.9% (SD) IS of the area at risk. A single cycle of 10-min preconditioning ischemia followed by 15-min reperfusion reduced IS after 90 min of ischemia to 2.8 +/- 2.7%. The epicardial monophasic action potential duration at 50% repolarization (MAP50) was decreased more markedly during the initial 10 min of the prolonged ischemia than during the first 10 min of the preconditioning ischemic period (84 +/- 4 vs. 89 +/- 2%). Transmural myocardial adenosine (ADO) uptake was reversed to net release during both ischemic periods and during the initial phase of reperfusion. Glibenclamide (0.5 mg/kg, followed by 50 micrograms/min i.v.) abolished the reduction in MAP50 without altering ADO release. Glibenclamide did not alter IS per se (13.0 +/- 7.6%) but abolished the beneficial effect of IP (IS: 13.6 +/- 6.2%). Thus blockade of KATP with glibenclamide abolishes the IS-reducing effect of IP in swine but does not reduce ADO release.

Action Potentials↗

Genetic susceptibility in familial multiple sclerosis not linked to the myelin basic protein gene.

The myelin basic protein (MBP) gene is a candidate locus for disease susceptibility in familial multiple sclerosis. Amplification of a polymorphic tetranucleotide repeat region immediately 5' to MBP exon 1 demonstrated the presence of eight different alleles among members of 14 multiplex multiple sclerosis families (36 affected individuals). Linkage analysis was performed with autosomal dominant and autosomal recessive models, normal individuals with abnormal magnetic resonance scans being scored as either unknown or affected. Cumulative LOD scores were negative for both models of inheritance. The results do not demonstrate linkage between the MBP gene region and multiple sclerosis.

Alleles↗

A comparison of oxygen pulse and respiratory exchange ratio in cerebral palsied and nondisabled children.

Energy expended while walking is increased for children with cerebral palsy compared to nondisabled children. This study compared oxygen uptake, oxygen pulse, and the respiratory exchange ratio (RER) in children with cerebral palsy and nondisabled children walking on a treadmill. Resting oxygen uptake and oxygen pulse values were not different in the two groups. At a given walking speed, oxygen uptake, oxygen pulse, and RER were higher for subjects with cerebral palsy. At a given level of submaximal oxygen uptake, oxygen pulse and RER values were not different in subjects with cerebral palsy compared to nondisabled children. It was concluded that the cardiorespiratory response to walking at submaximal level of work is not significantly different for children with cerebral palsy.

Adolescent↗

Prolactin binding sites in the adrenal glands of mink (Mustela vison).

1. The purpose of this study was to determine if the mink adrenal gland might be a target organ for prolactin by establishing whether or not binding sites for the hormone exist in adrenal cell membranes. 2. Adrenal glands were collected from adult female mink in November, 1991, homogenized and subjected to differential centrifugation into three particulate fractions; 1500, 15,000 and 50,000 g. All binding determinations were made using 125I-oPRL and 200-300 micrograms protein from the 50,000 g particulate fraction. Optimal binding occurred within 8 hr at 25 degrees C. 3. Scatchard analysis of saturation data revealed a single set of high affinity (Kd = 9.27 x 10(-11) +/- 1.63 M), low capacity (Bmax = 34.22 +/- 5.37 fmol/mg) binding sites. 4. Binding sites appeared to be hormone specific as only oPRL (73% displacement) and oLH (8% displacement) inhibited binding of 125I-oPRL to adrenal membranes. No inhibition of 125I-oPRL binding to adrenal membranes occurred in the presence of a 500-fold excess of bTSH, oGH or oFSH. 5. Prolactin binding sites were readily detected in adrenal and kidney tissue, but were low in liver and almost non-detectable in spleen or lung tissue. 6. Our data suggest that the mink adrenal gland is a target organ for prolactin and that an interaction between the pituitary and adrenal glands may exist that is important for the regulation of such physiological processes as fur growth cycles.

Adrenal Glands↗

Post-ejection wall thickening as a marker of successful short term hibernation.

OBJECTIVE: Short term hibernating myocardium is characterised by a decrease in contractile function in proportion to the reduced blood flow, the recovery of creatine phosphate despite ongoing ischaemia, a recruitable inotropic reserve, and the absence of necrosis. During acute myocardial ischaemia systolic wall thickening decreases and post-ejection wall thickening develops. The extent of post-ejection thickening during severe ischaemia correlates with the recovery of contractile function during reperfusion. Whether the extent of post-ejection wall thickening can also distinguish short term hibernating myocardium from more severely ischaemic, infarcting myocardium and thus predict the amount of viable tissue was tested in 13 anaesthetised pigs. METHODS: The left anterior descending coronary artery (LAD) was cannulated and perfused at constant flow. After control measurements of regional myocardial blood flow (with radiolabelled microspheres) and wall thickening (sonomicrometry), coronary inflow was reduced to produce a reduction in regional contractile function by 60-100%. After 85 minutes of ischaemia, dobutamine was infused into the LAD for five minutes to determine the extent of inotropic reserve. Transmural biopsies were taken to measure regional myocardial creatine phosphate content and infarct size was determined after two hours of reperfusion by staining with triphenyl tetrazolium chloride. RESULTS: The extent of post-ejection wall thickening after 85-90 minutes of ischaemia correlated with the myocardial creatine phosphate content (r = 0.812, n = 11, p < 0.01) and the extent of the dobutamine recruitable inotropic reserve (r = 0.783, n = 7, p < 0.05). A negative correlation existed between the extent of post-ejection wall thickening and % infarct size (r = -0.699, n = 10, p < 0.05 for the transmural piece of tissue containing the ultrasonic crystals; r = -0.743, n = 10, p < 0.05 for the area of the left ventricle at risk). Finally, post-ejection wall thickening after 85-90 minutes of ischaemia correlated with the recovery of contractile function at 30 minutes reperfusion (r = 0.657, n = 10, p < 0.05). CONCLUSION: The extent of post-ejection wall thickening may indicate the amount of viable tissue after 85-90 minutes of low flow ischaemia. The greater the post-ejection wall thickening, the more myocardium is successfully hibernating.

Animals↗

Role of ovarian steroids in development of uterine binding sites for prolactin in the ferret.

The objectives of this study were to investigate 1) the presence of specific prolactin (PRL) binding sites in the ferret uterus, 2) the uterine location of 125I-labeled ovine PRL (oPRL) binding sites, 3) changes in uterine PRL binding sites during pseudo-pregnancy, and 4) regulation of PRL binding sites by ovarian steroids. Binding was determined through use of 125I-oPRL and 300-800 micrograms of protein from the 50,000 x g particulate fraction. Optimal binding occurred within 6 h at 25 degrees C. Scatchard analysis of saturation data revealed a single set of high-affinity (Kd = 4.99 x 10(-11) +/- 0.88 M), low-capacity (22.76 +/- 1.62 fmol/mg) binding sites. Analysis of hormonal specificity revealed that ovine growth hormone (oGH) cross-reacted with oPRL for the uterine binding sites, displacing 38% of the bound ligand. However, no inhibition of 125I-oPRL binding occurred in the presence of a 500-fold excess of bovine thyroid-stimulating hormone (bTSH), ovine LH (oLH), or ovine FSH (oFSH), suggesting hormonal specificity of the binding sites that are located in the luminal and glandular epithelium. Prolactin binding to ferret uterine membranes increased during the first half of pseudopregnancy, plateaued between Days 21 and 28, and then declined. The concentration of PRL binding sites in uteri of ferrets on Day 1 of pseudopregnancy was 4.91 +/- 0.42 fmol/mg of protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Development of short-term myocardial hibernation. Its limitation by the severity of ischemia and inotropic stimulation.

BACKGROUND: Short-term hibernating myocardium is characterized by a decrease in contractile function in proportion to the reduced myocardial blood flow. Myocardial creatine phosphate content, initially decreased during the first minutes of ischemia, returns to near-control values, the ischemia-induced net lactate production is attenuated, and the myocardium remains viable despite ongoing hypoperfusion and contractile dysfunction. Hibernating myocardium after 85 minutes of ischemia maintains an inotropic reserve and responds to short-term intracoronary dobutamine infusion with increased work; however, this inotropic response is at the expense of metabolic recovery. We therefore hypothesized that the development of myocardial hibernation is a delicate process that is easily disturbed by unfavorable alterations in the oxygen-supply demand balance. METHODS AND RESULTS: To study the impact of prolonged inotropic stimulation on the development of myocardial hibernation, the left anterior descending coronary artery was cannulated and hypoperfused at constant flow in 12 enflurane-anesthetized swine. The reduction of coronary inflow was followed by a reduction of regional myocardial work (sonomicrometry) from 248 +/- 59 mm Hg.mm to 73 +/- 35 mm Hg.mm (P < .05) at 5 minutes of ischemia. Dobutamine (2.5 +/- 1 micrograms/min) was then infused for an additional 85 minutes. Work was increased at 5 minutes of dobutamine to 139 +/- 34 mm Hg.mm (P < .05 versus 5 minutes of ischemia). However, this increase was only transient, and after 85 minutes of dobutamine, work was decreased below the initial ischemic value (42 +/- 34 mm Hg.mm). At 5 minutes of ischemia, creatine phosphate content was reduced from 8.80 +/- 1.97 to 6.21 +/- 3.87 mumol/g wet wt, and myocardial ATP content was decreased slightly from 4.75 +/- 0.92 to 4.12 +/- 1.29 mumol/g wet wt (both, P = NS). After 5 minutes of dobutamine, further reductions in creatine phosphate content to 3.11 +/- 0.76 mumol/g wet wt and in ATP to 3.14 +/- 0.81 mumol/g wet wt were observed (both, P < .05 versus control). During the remainder of the continuous dobutamine infusion, creatine phosphate content remained unchanged, whereas ATP further decreased significantly to 1.68 +/- 0.96 mumol/g wet wt. The beta-adrenoceptor density of the left anterior descending coronary artery-perfused myocardium was 36.5 +/- 5.8 fmol (-)-[125I]iodocyanopindolol/mg protein under control conditions, and this was unchanged during ischemia and the subsequent dobutamine infusion. Following 90 minutes of ischemia with 85 minutes of dobutamine and 2 hours of reperfusion, infarct size (triphenyl tetrazolium chloride staining) was 26.3 +/- 7.5% of the area at risk. With constant hypoperfusion, dobutamine redistributed blood flow away from the subendocardium (0.20 +/- 0.08 versus 0.11 +/- 0.04 mL.min-1.g-1) toward the subepicardium (0.45 +/- 0.13 versus 0.51 +/- 0.21 mL.min-1.g-1) as well as to the right ventricle (0.26 +/- 0.08 versus 0.32 +/- 0.09 mL.min-1.g-1). Therefore, in two other groups of six and five swine, the severity of ischemia was increased to achieve an 80% or a 90% reduction in regional function, respectively, and the importance of the severity of blood flow reduction per se for the development of myocardial infarction was studied. The infarct size in the animals undergoing 85 minutes of dobutamine (26.3 +/- 7.5%) was increased above the level expected from the blood flow reduction alone (6.3 +/- 6.4%, P < .01). CONCLUSIONS: Both the increased severity of ischemia and the enhanced energy expenditure induced by dobutamine impair the development of myocardial short-term hibernation and precipitate myocardial infarction.

Animals↗

Nutrient needs of the preterm infant.

Preterm infants exhibit special nutrient needs that differ substantially from other patient populations. Special characteristics include increased energy and protein requirements that must be addressed to prevent tissue catabolism and support growth. The immaturity of some organ systems may also complicate the administration of nutrition support. This article describes important characteristics of enteral and parenteral nutrition as it applies to the preterm infant.

Enteral Nutrition↗