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Biomedical subjects

S Clark

Publications and source records attributed to S Clark.

At least 253 records · Page 14Linked to original sources
PubMed1991

A convenient multiplex PCR system for the detection of dystrophin gene deletions: a comparative analysis with cDNA hybridisation shows mistypings by both methods.

Existing reactions for the multiplex PCR amplification of exons in the dystrophin gene have been modified to produce two multiplex reactions which separately cover the 5' and 3' major deletion 'hotspots' in the gene, and together detect approximately 98% of all deletions detectable by Southern cDNA hybridisation. A comparative study of 148 patients showed mistypings in both the cDNA hybridisation data (4%) and the PCR analysis (1.2%). We suggest means of circumventing the underlying problems in order to avoid mistyping and subsequent misdiagnosis, and conclude that, with appropriate precautions, multiplex PCR amplification can be the method of choice for detecting deletions in the dystrophin gene.

Base Sequence↗
PubMed1991

Sequential administration of interleukin-6 and granulocyte-colony stimulating factor in newborn rats: modulation of newborn granulopoiesis and thrombopoiesis.

During states of increased demand, neonatal host defense is characterized by dysregulation of granulopoiesis, resulting in a high incidence of neutropenia. This study investigated the modulation of neonatal rat hematopoiesis by 14-d administration of recombinant human (rh) IL-6, rh-granulocyte-colony stimulating factor (G-CSF), or sequential combination of rhIL-6 and rhG-CSF. Specifically, newborn Sprague-Dawley rats were treated with either rhIL-6 (5 micrograms/kg/d for 14 d), rhG-CSF (5 micrograms/kg/d for 14 d), rhIL-6 for 7 d followed by rhG-CSF for 7 d, PBS/BSA for 7 d followed by rhG-CSF for 7 d, or PBS/BSA for 14 d. RhIL-6 alone significantly increased the peripheral platelet count during the latter part of the 2nd wk of administration (d 13: 980 +/- 42 versus 716 +/- 23 x 10(3)/mm3) (p = less than 0.001) (mean +/- SEM). Treatment with rhIL-6 for 7 d followed by rhG-CSF significantly increased the peripheral neutrophil count compared with 7 d of PBS/BSA and 7 d of G-CSF (d 14 absolute neutrophil count 4888 +/- 12 versus 2720 +/- 317/mm3) (p = less than 0.05). Similarly, sequential rhIL-6/rhG-CSF significantly increased the d-14 bone marrow neutrophil storage pool (9873 +/- 882 versus 3564 +/- 159/mm3) (p = less than 0.005). Lastly, sequential rhIL-6/rhG-CSF induced the highest increase in bone marrow (p less than 0.01) and liver/spleen CFU-GM pool (p less than 0.001) compared with any other treatment group. These studies suggest that rhIL-6 alone is associated with a significant increase in the neonatal platelet count.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Multifactor stimulation of megakaryocytopoiesis: effects of interleukin 6.

We have previously demonstrated that interleukin 3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF) stimulate various aspects of megakaryocytopoiesis. We have investigated the capacity of interleukin 6 (IL-6) to stimulate megakaryocyte colony formation from both normal Balb/C marrow and light-density marrow extensively depleted of adherent, pre-B, B and T cells. Human recombinant IL-6 (167 ng/ml) stimulated megakaryocyte colony formation from normal marrow (8.6 +/- 1 megakaryocyte colony-forming units [CFU-meg]/10(5) cells) as compared to control (1.5 +/- 4 CFU-meg/10(5) cells) in 16 determinations (p less than 0.01). IL-6 (167 ng/ml) also stimulated CFU-meg formation from depleted marrow (control, 10.8 +/- 4 CFU-meg/10(5) cells versus IL-6, 68 +/- 19 CFU-meg/10(5) cells in 12 determinations, p less than 0.01). IL-6 synergistically augmented IL-3-induced colony formation (139% IL-3 control, 120% calculated IL-3 plus IL-6 control, n = 11, p less than 0.01) in normal marrow and showed an additive effect in depleted marrow (133% IL-3 control, p less than 0.01, 114% of IL-3 plus IL-6, value not significant [NS] at 0.05 level). Studies with recombinant murine IL-6 gave similar results. There was an increasing level of megakaryocyte colony-stimulating activity from G-CSF (16,667 U/ml, 2.47 +/- 0.6 CFU-meg/10(5) cells, n = 17), to IL-6 (167 ng/ml, 8.47 +/- 0.96 CFU-meg/10(5) cells, n = 19), to GM-CSF (52 U/ml, 23 +/- 4 CFU-meg/10(5) cells, n = 14), to IL-3 (167 U/ml, 48 +/- 5 CFU-meg/10(5) cells, n = 20) as compared to media-stimulated marrow (range 1.29-1.86 CFU-meg/10(5) cells). A similar hierarchy was seen with depleted marrow. Combinations of factors (including IL-3, GM-CSF, G-CSF, and IL-6) tested against normal unseparated murine marrow did not further augment CFU-meg numbers over IL-3 plus IL-6 but did increase colony size. These data suggest that IL-6 is an important megakaryocyte regulator, that at least four growth factors interact synergistically or additively to regulate megakaryocytopoiesis, and that combinations of growth factors, possibly in physical association, might be critical in stimulating megakaryocyte stem cells.

Animals↗
PubMed1991

Signs, symptoms, and the new descriptors.

Data from 143 consultations in general practice were assessed by factors relevant to the four-category descriptive system developed by the Royal Australian College of General Practitioners. It seems that there is no relationship between consultation length, apparent symptom seriousness or the occurrence of a physical examination. Suggestions are offered for improving the clarity of consultation descriptor categories.

Diagnosis↗
PubMed1991

Ambulatory blood pressure monitoring during pregnancy: validation of the TM-2420 monitor.

To validate the accuracy of the TM-2420 ambulatory blood pressure monitor in pregnant women, its measurements were compared with those taken simultaneously by two trained observers using a random zero sphygmomanometer. Each of 30 pregnant women had her blood pressure measured three times. The mean differences between the monitor and the averaged observer measurements were -0.53 +/- 2.7 mmHg for systolic blood pressure, -5.4 +/- 5.3 mmHg for Korotkoff phase 4 (P less than .001), and 0.87 +/- 3.7 mmHg for phase 5 diastolic blood pressure. The TM-2420 ambulatory monitor provides reliable estimates of systolic and Korotkoff phase 5 diastolic blood pressures during pregnancy.

Blood Pressure Monitors↗
PubMed1991

Case management.

Explore the source record for details and available documents.

Adult↗
PubMed1991

Ambulatory blood pressure monitoring: validation of the accuracy and reliability of the TM-2420 according to the AAMI recommendations.

The accuracy of the TM-2420 ambulatory blood pressure monitor (A&D Co, Japan) has been assessed by the indirect method according to the recommendations of the Association for the Advancement of Medical Instrumentation (AAMI). Ninety subjects (43 men and 47 women), aged 19-89 with a range of systolic blood pressures (SBP) of 81-211 mmHg and diastolic blood pressures (DBP) of 32-113 mmHg were studied. The monitor was compared with two observers using the Hawksley random zero sphygomomanometer. The standard deviation of the difference (SDD) between the observers was 3.0 mmHg for SBP and 2.3 mmHg for DBP. The mean differences between observers were -0.31 mmHg for SBP and 0.32 mmHg for DBP (both NS). The average of three readings for each subject recorded by the monitor was compared with the average of the simultaneous readings by the observers. The SDD was 7.2 mmHg for SBP and 5.5 mmHg for DBP (within the recommendations for accuracy). The differences between methods were -0.98 mmHg for SBP and 0.18 mmHg for DBP; these differences were not significant (NS). The monitor was also assessed against direct intra-brachial artery pressure in 12 subjects (36 readings). The mean difference between the monitor and simultaneous individual intra-arterial reading was -9.5 mmHg for SBP (P less than 0.001) and 3.7 mmHg for DBP (P less than 0.001). The SDD's between methods were 12 mmHg for SBP and 5.0 mmHg for DBP. Use of the monitor in general clinical practice in 100 patients was also assessed. The rate of errors was low (6.8%), and the device found to be acceptable.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Expression of epidermal-growth-factor receptor in the K562 cell line by transfection. Altered receptor biochemistry.

The epidermal-growth-factor (EGF) receptor was expressed in the human erythroleukaemic cell line K562 by transfection of the receptor cDNA. EGF-receptor biochemistry appears altered in the K562 transfectants. Autophosphorylation of the K562 receptor is not stimulated substantially by EGF. Tyrosine kinase activity of the receptor is high in the absence of EGF, whereas receptor affinity for EGF is low. K562 cells are shown to lack mRNA for transforming growth factor alpha (TGF alpha). Therefore autocrine stimulation of the K562 receptor, at least by TGF alpha, does not explain the observed receptor biochemistry. The K562 receptor is phosphorylated at a single major site in intact cells, a threonine residue that may be Thr-669. Possible mechanisms of regulation of the EGF receptor in the K562 transfectants are discussed.

DNA↗
PubMed1990

Splice junction mutations in a yeast tRNA gene which alter the rate and precision of processing.

We have introduced mutations into a tRNALeu3 gene which alter the intron boundaries and examined their effects on RNA splicing. Our results show that the 5'-proximal splice junction is not specified by the position of an adjacent base-paired stem present in all naturally occurring tRNA precursors. Also, efficient cleavage of 5'-splice junctions unique to these mutants, -CpU-, -UpA- and -UpG-, indicates the purine found at the 5'-side of this site in all natural precursors is dispensable. Some alterations of the sequence and structure at the 5'-proximal splice site reduce the rate of cleavage therein and result in accumulation of molecules composed of the 5'-half of the tRNA plus the intron. The precise position of the 5'-proximal cleavage site can vary +/- 1 base in these mutants. The 3'-proximal splice junction is rendered inactive by changing the prospective splice junction sequence from -ApC- to -CpC- and reducing the size of an unpaired loop at this site from six to two bases. Very small amounts of RNA composed of the 3'-half of the tRNA plus the intron accumulate from this precursor. We conclude that splice junction sequence and structure affect both the rate and precision of intervening sequence removal.

Base Sequence↗
PubMed1990

Nicardipine versus placebo for the treatment of postoperative hypertension.

Postoperative hypertension can cause serious complications, including bleeding from fresh anastomoses, cardiovascular accident, and myocardial ischemia. Therefore rapid control of blood pressure is essential to prevent poor outcome. In this study, 30 American Society of Anesthesiologists class I and II patients who did not have cardiac surgery and subsequently developed postoperative hypertension were randomly assigned to receive either nicardipine, a new dihydropyridine calcium channel blocker, or placebo. Intravenous nicardipine was given as a loading bolus of 10 mg/hr for 5 minutes and was titrated to 15 mg/hr if needed to achieve a therapeutic response. After therapeutic response, intravenous nicardipine was decreased to 3 mg/hr and subsequently titrated in increments of 1.0 to 2.5 mg/hr to maintain blood pressure control. Systolic and diastolic blood pressures during titration and maintenance did not differ significantly from preoperative levels in patients treated with nicardipine. The mean time to therapeutic response for the nicardipine-treated group was 8.67 +/- 1.46 minutes, and the median time to offset of action was 15 minutes. Eleven of the 12 patients who received placebo were crossed over to antihypertensive therapy, and of these, 10 received intravenous nicardipine. In this group all achieved therapeutic response in 7.3 +/- 1.18 minutes. The usefulness of intravenous nicardipine for postoperative hypertension was demonstrated in this study by: (1) the rapid control of blood pressure, (2) its continued efficacy during maintenance, and (3) little need to adjust dosage to control blood pressure.

Dose-Response Relationship, Drug↗
PubMed1990

Strand-specific mismatch correction in nuclear extracts of human and Drosophila melanogaster cell lines.

Nuclear extracts derived from HeLa and Drosophila melanogaster KC cell lines have been found to correct single base-base mispairs within open circular DNA heteroduplexes containing a strand-specific, site-specific incision located 808 base pairs from the mismatch. Correction in both extract systems is strand specific, being highly biased to the incised DNA strand. Different mispairs within a homologous set of heteroduplexes were processed with different efficiencies (G.T greater than G.G approximately equal to A.C greater than C.C), and correction was accompanied by mismatch-dependent DNA synthesis localized to the region spanning the mispair and the strand break, thus demonstrating that mismatch recognition is associated with the repair reaction. Correction of each of these heteroduplexes was abolished by aphidicolin but was relatively insensitive to the presence of high concentrations of ddTTP, indicating probable involvement of alpha and/or delta class DNA polymerase(s). These findings suggest that higher eukaryotic cells possess a general, strand-specific mismatch repair system analogous to the Escherichia coli mutHLS and the Streptococcus pneumoniae hexAB pathways, systems that contribute in a major way to the genetic stability of these bacterial species.

Animals↗
PubMed1990

Nursing interventions for the depressed cardiovascular patient.

Patients who have a cardiovascular illness may experience sad feelings that range from normal grief reactions to major depressive episodes. Unresolved depression has been shown to affect long-term recovery and mortality. This article explores the differences in these depressive responses so that nurses will be able to focus their assessments and identify appropriate interventions.

Cardiovascular Diseases↗
PubMed1990

Anti-sperm antibodies in homosexual men: prevalence and correlation with sexual behaviour.

The sera of 60 homosexual males were examined for the presence of antibodies to sperm using an indirect immunobead test (IBT). Six of 60 (10%) had antibodies of IgG isotype; in addition two of the six had antibodies of IgA isotype. The presence of antibodies was associated with the practice of unprotected receptive anal intercourse in the previous six months. Antibodies were not found in homosexual men who were celibate, or who practised only oral intercourse during the same period. There was no correlation between the presence of anti-sperm antibodies and antibodies to human immunodeficiency virus (HIV), or numbers of T lymphocytes. These preliminary results lend support to the hypothesis that antigen presentation in the lower gut may be a source of sensitisation against sperm. The possibility that anti-sperm antibodies may be a marker of receptive anal intercourse merits further investigation.

Adult↗
PubMed1990

Expression of atypical and classical insulin receptors in Chinese hamster ovary cells transfected with cloned cDNA for the human insulin receptor.

Human placenta and IM-9 lymphocytes contain subpopulations of atypical insulin receptors which differ from classical insulin receptors in their higher binding affinity for insulin-like growth factors I and II (IGF-I and IGF-II). Both types of insulin receptors may be derived from different but related genes, or may represent alternative post-translational modifications of the same gene product. To test these possibilities, we have examined the IGF binding characteristics of the human insulin receptors expressed in Chinese hamster ovary (CHO) cells which had been stably transfected with cloned human insulin receptor cDNA (CHO-T cells). The parent CHO cells contained 3 x 10(3) rodent insulin receptors/cell, and the CHO-T cells, 2.0 x 10(6) human insulin receptors/cell. Competition binding studies showed that the binding of [125I]IGF-I and [125I]multiplication stimulating activity (MSA/rat IGF-II) to parent CHO cells was primarily to type I and II IGF receptors, which cross-react poorly or not at all with insulin. However, competition binding studies with CHO-T cells showed that [125I]IGF-I binding was displaced 60-70%, and [125I]MSA binding, 50-55%, by low concentrations of insulin (20 ng/ml) and no further by higher concentrations of insulin (500 ng/ml). The insulin-insensitive IGF binding sites corresponded to the rodent type I and II IGF receptors; the insulin-sensitive IGF binding sensitive sites resembled the human atypical insulin receptors in that they bound IGF-I and MSA with moderately high affinity and reacted with insulin, MSA, and IGF-I in that order of potency. Atypical insulin receptors were also demonstrated by insulin-sensitive [125I]IGF-I and [125I]MSA binding to solubilized CHO-T proteins adsorbed to microtiter wells coated with monoclonal antibodies specific for the human insulin receptor. These results suggest that atypical human insulin receptors are generated by differential post-translational processing of the same gene product as classical human insulin receptors.

Animals↗
PubMed1989

NMDA antagonists differentiate epileptogenesis from seizure expression in an in vitro model.

In an electrographic model of seizures in the hippocampal slice, both of the N-methyl-D-aspartate (NMDA) antagonists 2-amino-5-phosphonovaleric acid and 5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine maleate (MK-801) prevented the progressive development of seizures but did not block previously induced seizures. Thus, a process dependent on the NMDA receptor-ionophore complex establishes a long-lasting, seizure-prone state; thereafter the seizures depend on non-NMDA receptor-ionophore mechanisms. This suggests that there is an important distinction between epileptogenesis and seizure expression and between antiepileptogenic and anticonvulsant pharmacological agents.

2-Amino-5-phosphonovalerate↗
PubMed1989

Proteolytic generation of constitutive tyrosine kinase activity of the human insulin receptor.

Structural modification induced by partial digestion with trypsin has been shown to stimulate the tyrosine kinase activity of the insulin receptor both in solution and in intact cells [Tamura, Fujita-Yamaguchi & Larner (1983) J. Biol. Chem. 258, 14749-14752; Goren, White & Kahn (1987) Biochemistry 26, 2374-2382; Leef & Larner (1987) J. Biol. Chem. 262, 14837-14842]. Furthermore, experiments involving deletion of sequences encoding the extracellular domain of the insulin receptor suggest that it may function as a protooncogene in fibroblasts [Wang et al., (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 5725-5729]. To further understand the structural requirements that generate this activity, the major activated fragments generated in solution following trypsin digestion have been characterized here, one of which is shown to have a similar amino acid sequence to a transforming protein. Furthermore, treatment with trypsin of intact Chinese hamster ovary cells that overexpress the human insulin receptor stimulates both autophosphorylation of the receptor and 2-deoxyglucose uptake into the cells, but does not enhance receptor internalization. Unlike digestion in solution, no proteolysis or loss of activity of the activated insulin receptor beta-subunit could be detected using intact cells, even at high trypsin concentrations, despite the existence of extracellular sites that are readily cleaved by trypsin in the solubilized receptor. These studies provide further detail of a mechanism used during trypsinization of cells in culture which mimics activation of the insulin receptor and contributes to stimulation of growth.

Amino Acid Sequence↗
PubMed1989

Interference of automated electroencephalographic processing by an endocardial pacemaker.

This article reports a patient with an endocardial pacemaker undergoing a carotid endarterectomy in whom cerebral function was monitored by using processed electroencephalography (EEG). The EEG was processed by means of aperiodic analysis. The pacemaker generated an artifact that was identified on the processed EEG display and on the raw EEG display. During the time of carotid cross-clamping, a loss of EEG activity was noted on careful examination of the processed EEG and confirmed by referring to the raw EEG. Nevertheless, because of the presence of the artifact, the activity edge remained essentially constant despite the change in underlying EEG activity. The cross-clamp was released based on the observed change in EEG activity, yielding a return to baseline EEG activity. The surgeon elected to shunt the patient. The patient awoke without neurologic sequelae. This case serves to demonstrate the importance of referring to the raw EEG signal and the limitations of the activity edge in the presence of artifact.

Carotid Artery Diseases↗