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K M Taylor

Publications and source records attributed to K M Taylor.

At least 163 records · Page 9Linked to original sources
PubMed1994

A method for in vitro synthesis of unglycosylated recombinant complement component C9.

A method for in vitro synthesis of human complement component C9 has been established in order to generate unglycosylated normal and mutant proteins without the need to sub-clone. One or two step polymerase chain reaction (PCR) was used to add the T7 RNA polymerase promoter and introduce multiple mutations within the cDNA. The cDNA was then transcribed by T7 RNA polymerase and the mRNA translated in a rabbit reticulocyte lysate or wheat germ system. Successful synthesis was confirmed by: the correct size of PCR product DNA on agarose gel electrophoresis, incorporation of [alpha-32P]UTP into mRNA, and formation of [35S]methionine-labelled protein of the correct molecular mass for full length C9. The wheat germ extract generated up to 1.5 micrograms of recombinant C9. This unglycosylated C9 had at least 10% of the haemolytic activity of native C9. Unglycosylated C9 polymerised more readily than the native protein. This spontaneous polymerisation was increased by removal of the first 23 amino acids or mutating two cysteines at positions 33 and 36. This therefore provides a rapid method for screening the effect of multiple mutations on the biological activity and polymerisation of pore forming proteins.

Animals↗
PubMed1994

Left ventricular function and coronary artery disease progression early after coronary bypass grafting.

To investigate the effects of coronary artery disease progression on left ventricular function in patients who suffer angina early after coronary artery bypass grafting, we studied the progression of coronary stenoses, the occurrence of graft occlusions, and measured left ventricular ejection fraction (regional and global) in 34 consecutive patients who underwent repeat angiography 25.2 +/- 3.5 (standard error of the mean) months postoperatively, from a total population of 550 patients who underwent bypass grafting. Resting left ventricular function and stenosis severity were assessed using a computerized, quantitative analysis system. Coronary stenosis progression was defined as an increase in the percentage of the stenotic occlusion by 30% or more, any increase in lesion severity that resulted in total coronary artery occlusion, or the occurrence of a new stenosis that occluded the artery by 50% or more. Group 1 comprised 21 patients with all grafts patent and group 2 comprised 13 patients with one or more grafts occluded (20 of 34 grafts). Coronary artery disease progressed in all patients in group 1, and this involved 22 of 54 (41%) grafted vessels and 3 of 15 (20%) nongrafted vessels (p < 0.05). Coronary artery disease progressed in 11 patients in group 2, involving 15 of 32 (47%) grafted vessels and 1 of 6 (17%) nongrafted vessels (p < 0.01). An increased collateral circulation was observed in both groups. The left ventricular ejection fraction remained unchanged in both groups (group 1, 0.60 +/- 0.03 versus 0.62 +/- 0.03; group 2, 0.62 +/- 0.05 versus 0.62 +/- 0.04 before and after bypass, respectively; p = not significant) and there was no difference between the groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Effect of cardiopulmonary bypass on gastrointestinal perfusion and function.

Gastric mucosal tonometry was used to determine the adequacy of gastrointestinal perfusion in 10 patients undergoing elective myocardial revascularization. Patients were prospectively randomized to receive either pulsatile or nonpulsatile flow during cardiopulmonary bypass. All patients showed a reduction in gastric mucosal perfusion during bypass, manifested by a reduction in the gastric mucosal pH, which occurred independently of variations in the arterial pH. In the group of patients receiving nonpulsatile flow, this reduction was significantly greater (p < 0.05). Cardiopulmonary bypass using nonpulsatile flow is associated with the development of a gastric mucosal acidosis, which may have implications for the development of postoperative complications.

Acidosis↗
PubMed1994

Effects of cardiopulmonary bypass on gut blood flow, oxygen utilization, and intramucosal pH.

Studies documenting rises in endotoxin after cardiopulmonary bypass (CPB) have postulated gut mucosal hypoperfusion. We have investigated alterations in jejunal blood flow by laser Doppler flow measurement, intramucosal pH (pHi) by tonometry, and oxygen utilization in a canine model of hypothermic CPB (n = 11 dogs). After 10 minutes of hypothermic CPB, despite no major reduction in superior mesenteric artery flow, mucosal laser Doppler flow decreased to -38.2% +/- 9.3% of levels obtained before bypass (p = 0.008) and serosal laser Doppler flow, to -47.3% +/- 11.4% (p = 0.006). During the hypothermic phase, mesenteric oxygen consumption fell from 0.18 +/- 0.01 to 0.098 +/- 0.01 mL.min-1.kg-1 (p = 0.005), and mesenteric oxygen delivery fell from 1.97 +/- 0.39 to 1.14 +/- 0.12 mL.min-1.kg-1 (p = 0.05). There was no change in jejunal pHi. During the rewarming phase, there was a substantial increase in mucosal laser Doppler flow, peaking at +69.8% +/- 15.2% (p = 0.03), whereas serosal laser Doppler flow returned to values seen prior to CPB (-16.4% +/- 21.5%; p = 0.25). These changes coincided with a surge in oxygen consumption (0.33 +/- 0.042 mL.min-1.kg-1; p = 0.009), while mesenteric oxygen delivery remained depressed at 1.09 +/- 0.12 mL.min-1.kg-1 (p = 0.04). Jejunal pHi fell from a value of 7.36 +/- 0.04 before CPB to 7.12 +/- 0.07 (p = 0.02), thus indicating mucosal hypoxia. During the rewarming phase of hypothermic CPB, there is a disparity between mesenteric oxygen consumption and oxygen delivery with villus tip ischemia; these findings may explain the pathophysiology of endotoxemia during CPB.

Animals↗
PubMed1994

Perioperative histologic and ultrastructural changes in the pericardium and adhesions.

The presence of pericardial adhesions prolongs the operation time and increases the risk of serious damage to the heart and other major vascular structures during resternotomy. The reported incidence of such damage is 2% to 6%. Pericardial mesothelial cells exhibit fibrinolytic activity, and therefore have an actual or potential role in the breakdown of the fibrinous adhesions that serve as the initial scaffolding for the firm collagenous adhesions seen at reoperation. Ten patients undergoing primary cardiac procedures were studied to assess the morphologic changes that take place within the pericardium and to relate these to accompanying changes in the pericardial plasminogen activating activity. Samples were taken at 0, 75, and 135 minutes after pericardiotomy. Compared with samples obtained at the time of pericardiotomy, those taken at 75 and 135 minutes demonstrated a significant progression in the mesothelial cell damage (p < 0.01), together with increasing evidence of pericardial inflammation (p < 0.01). The findings from electron microscope studies confirmed and supplemented these findings. Furthermore, compared with its initial levels (median, 2.06 IU/cm2; range, 1.28 to 6.48 IU/cm2), the plasminogen activating activity of pericardial biopsy specimens was significantly reduced at 75 minutes (median, 0.64 IU/cm2; range, 0.12 to 2.44 IU/cm2; P < 0.05), with some recovery at 135 minutes (median, 1.45 IU/cm2; range, 0.12 to 4.39 IU/cm2; p = 0.059). This study has revealed that, during cardiac procedures, the pericardium undergoes inflammatory changes with concomitant damage to its mesothelium, together with a reduction in the pericardial mesothelial fibrinolytic potential.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

The effect of intestinal hypoperfusion on intestinal absorption and permeability during cardiopulmonary bypass.

BACKGROUND/AIMS: Mean arterial pressure is reduced during hypothermic cardiopulmonary bypass. The aim of this study was to assess whether this was associated with intestinal hypoperfusion and whether it affected intestinal absorption and permeability. METHODS: Twenty-six patients undergoing coronary artery bypass grafting underwent an intestinal absorption-permeability test involving ingestion of 3-O-methyl-D-glucose, D-xylose, L-rhamnose, and lactulose. Ingestion took place 2 days before, within 3 hours, and 5 days after hypothermic cardiopulmonary bypass. Hemodynamic parameters and gastric mucosal laser Doppler blood flow were measured perioperatively in eight patients. RESULTS: Hypothermic (28 degrees C), nonpulsatile cardiopulmonary bypass resulted in a 25% reduction in mean blood pressure, 10% reduction in cardiac index, and a 46% reduction in gastric mucosal laser Doppler blood flow. There was 85.4%, 85.5%, and 73.6% reduction (P < 0.01) in active (3-O-methyl-D-glucose) and passive (D-xylose) carrier-mediated transport and passive, nonmediated transcellular (L-rhamnose) transport in the immediate postoperative period, respectively. The differential urine excretion of lactulose/L-rhamnose increased sixfold. All parameters returned to control levels by the fifth postoperative day. CONCLUSIONS: Cardiopulmonary bypass, while maintaining generally acceptable levels of hemodynamic performance, is associated with significant intestinal hypoperfusion and malabsorption of monosaccharides, which may have implications for enteral drug treatment in the immediate postoperative period.

Adult↗
PubMed1994

Cerebral complications after coronary bypass grafting.

As the mortality associated with coronary artery bypass grafting has fallen, morbidity has become an increasing concern both by the medical profession and the general public. Postoperative quality of life, including the socioeconomic concern of returning to gainful employment, is of supreme importance. It is in this regard that brain injury following coronary artery bypass grafting is devastating if it leaves the patient with persistent motor, sensory, behavioral, or intellectual deficiencies. This paper reviews the cerebral complications following surgery for coronary artery disease. The extent of the problem is defined, and pathological mechanisms of injury together with methods employed to investigate cerebral pathology and physiology are reviewed. Finally, interventions which have been and are currently employed in an attempt to reduce cerebral injury are discussed.

Brain Diseases↗
PubMed1994

Myelodysplasia.

Myelodysplasia or myelodysplastic syndromes represent a heterogeneous group of bone marrow disorders characterized by dysmaturation, cytopenias, and a propensity for leukemic transformation. Although universally adopted, the French-American-British classification still has several limitations and an inability to categorize all patients. Refinements in morphologic and histologic interpretation in addition to the use of scoring systems may improve diagnostic and prognostic capability. Cytogenetics and molecular genetic abnormalities are providing clues to the fundamental pathogenesis of myelodysplastic syndromes. However, the lesions responsible for initiation or disease progression are as yet unresolved. Although chemotherapy and allogeneic transplantation may be used in selected patients, the mainstay of therapy remains supportive care, with differentiating therapy being largely disappointing so far and the role of hematopoietic growth factors remaining unresolved.

Chromosome Aberrations↗
PubMed1994

Magnetic resonance imaging and neuropsychological changes after coronary artery bypass graft surgery: preliminary findings.

Cerebral morbidity is a problem after cardiac surgery. Although neuropsychological tests and imaging techniques have been applied to cardiac patients, the relationship between them has not been considered. In the preliminary investigation, we studied 15 patients (11 male, mean age 59 years) having coronary artery bypass graft (CABG) surgery. Before surgery, patients had magnetic resonance (MR) imaging and neuropsychological assessment with a battery of 10 tests. During surgery, cardiopulmonary bypass was maintained at 28 degrees C with a flow rate of 2.4 L/m2/min-1 and at a mean arterial pressure of 50-70 mm Hg. Bubble or membrane oxygenators with in-line filters were used. Arterial blood gases were maintained using a pH-stat protocol. Fourteen of the 15 patients showed MR abnormalities before surgery. One week after surgery, four patients had additional MR changes. Six patients had significant postoperative neuropsychological deficit in memory (verbal and nonverbal) and attention. The four patients with new MR abnormalities all had significant neuropsychological deficit. In addition to corroborating previous observations that a high proportion of patients undergoing elective CABG have MR abnormalities before surgery, these preliminary data suggest a promising concordance between structural brain changes and cerebral function after CABG.

Adult↗
PubMed1994

Effect of prostacyclin PGI2 on cardiopulmonary bypass-induced lung injury.

Lung injury produced by cardiopulmonary bypass (CPB) is clinically characterized as postperfusion pulmonary dysfunction syndrome. The roles of humoral factors, altered perfusion modes and the occurrence of diffuse microembolism have been subjects of a number of studies. This paper presents the effectiveness of a platelet inhibiting drug, PGI2 in preventing occlusive microaggregates in the pulmonary circulation. In a series of experimental dog studies using a PGI2 dosage protocol of 10 ng/kg/minute for 30 minutes prior to the onset of CPB followed by 20 mg/kg/minute during CPB, the following effects have been observed: 1) Preservation of platelet numbers during CPB (p < 0.01 versus controls; n = 16). 2) Significant reduction in platelet aggregation during CPB (p < 0.01; n = 16). 3) Insignificant hypotensive effect at normal levels of peripheral vascular resistance (n = 16). 4) Occlusive fibrin, leucocytes and small platelet-based microaggregates obstructing pulmonary arterioles in six of the seven control dogs but in none of the dogs receiving PGI2 infusion. 5) No evidence of perivascular or intra-alveolar oedema, interstitial inflammatory cell infiltrates or haemorrhage was seen in either group of dogs. The controversy existing in relation to the possible therapeutic role of PGI2 and, in particular, its ability to prevent occlusive microaggregates in the arterioles and capillaries of vital organs should encourage further clinical studies of PGI2 and its derivatives during cardiac surgery.

Animals↗
PubMed1994

The effect of cardiopulmonary bypass on gastric and colonic mucosal perfusion: a tonometric assessment.

In a study to assess the potential effect of nonpulsatile hypothermic cardiopulmonary bypass (CPB), intramucosal pH (pHi) of the gastric and colonic mucosae was determined by tonometry (n = 8). During the hypothermic phase of CPB, gastric and colonic pHi did not change significantly. Forty minutes after the start of rewarming, despite increases in the cardiac index and mean arterial blood pressure, gastric pHi fell from 7.53 +/- 0.02 to 7.31 +/- 0.03 (p = 0.017) and colonic pHi fell from 7.50 +/- 0.02 to 7.32 +/- 0.03 (p = 0.028). Forty minutes after the end of CPB both the colonic (p = 0.017) and gastric (p = 0.046) pHi remained depressed below pre-CPB values. The difference in the arterial (pHa) and the gastric mucosal pH changed from -0.097 before CPB to 0.016, 40 minutes after the end of CPB (p = 0.027). This alteration in the pHa-pHi underlines the importance of measuring intramucosal pH by tonometry, since the pHa and pHi may move in opposite directions during episodes of haemodynamic stress. Both the gastric and colonic pHi were found to have a linear correlation with the pHa, although changes in the gastric pHi (r = 0.41, p = 0.018) were more strongly correlated with the pHa than the colonic pHi (r = 0.23, p = 0.19) in the rewarming phase of CPB and the immediate post-CPB period when there was a tendency towards intramucosal acidosis. The development of intramucosal acidosis in the rewarming and immediate post-CPB phases following hypothermic nonpulsatile CPB may impair the gut barrier and predispose patients to the absorption of luminal toxins.

Blood Pressure↗
PubMed1994

Fundamental dilemmas of the randomized clinical trial process: results of a survey of the 1,737 Eastern Cooperative Oncology Group investigators.

PURPOSE: We studied oncologists' attitudes and behavior with regard to their participation in randomized clinical trials. METHODS: We surveyed the 1,737 physician members of the Eastern Cooperative Oncology Group (ECOG) using the Physician Orientation Profile (POP), a self-administered mailed questionnaire. A response rate of 86% was achieved (1,485 of 1,737); each physician's actual patient accrual was recorded. RESULTS: All respondents indicated that they had a systematic pattern of patient preselection for entry onto trials beyond the formal inclusion/exclusion trial criteria. Eighty-nine percent stated that improving patient quality of life rather than prolonging survival was more personally satisfying. Sixty-two percent did not enter a single patient during the 12-month period following the survey, while 10% entered 80% of all patients during that time. Physicians overestimated their accrual rate by a factor of 6. Eighty-three percent defined randomization and adherence to trial protocol as a serious challenge to their ability to make individualized treatment decisions. CONCLUSION: This study raises questions regarding the following: (1) the perceived generalizability of trial findings, (2) the role of end points other than survival for clinical trials, (3) the consequences of physician overestimation of patient accrual, and (4) the impact of randomized trials on the behavior of clinicians. Further investigation into these critical issues will provide meaningful recommendations to enhance the future design, implementation, and conduct of randomized clinical trials in cancer.

Adult↗
PubMed1994

Altered glycosylation and selected mutation in recombinant human complement component C9: effects on haemolytic activity.

Recombinant wild-type and mutated forms of human complement component C9 have been synthesized in baculovirus-infected insect cells. Wild-type recombinant C9 was indistinguishable from native C9, as judged by haemolytic activity, trypsin and alpha-thrombin digestion, reaction with antibodies to C9, enzymatic deglycosylation to the same core size and polymerization in the presence of Zn2+. Replacement of the native signal peptide with the honey-bee melittin signal peptide, and replacement of Spodoptera frugiperda (Sf9) cells with Trichoplusia ni cells produced yields of 5 micrograms C9/ml supernatant. Three C9 mutants were generated; one mutant, with four acidic residues changed to alanines in a putative calcium-binding site, had the same biological activity as recombinant C9. Another mutant, lacking 23 N-terminal amino acids, previously showing increased polymerization when produced in vitro, polymerized on secretion, rendering it inactive. It was not possible to demonstrate haemolytic activity of the third mutant, cysteines 33 and 36 mutated to alanine, as it was secreted a hundredfold less than the wild-type protein.

Animals↗
PubMed1994

Co-expression of renin-angiotensin system component genes in human atrial tissue.

OBJECTIVES: To detect co-expression of genes coding for components of the renin-angiotensin system and investigate the potential for variation in the level of angiotensin converting enzyme (ACE) gene expression in the right atrial appendage of patients undergoing heart surgery. DESIGN: The right atrial appendage was collected at the time of surgery from 30 randomly chosen patients and was rapidly frozen in liquid nitrogen prior to extraction of messenger (m)RNA. Surgical samples of heart valve (n = 6) and papillary muscle (n = 3) were also examined. METHODS: Aliquots of purified mRNA were reverse-transcribed for analysis of gene expression by a polymerase chain reaction amplification assay. Primers specific for angiotensinogen, renin, ACE, cardiac chymase, atrial natriuretic peptide, glyceraldehyde-3-phosphate dehydrogenase, adenosine deaminase and the transferrin receptor were used for a qualitative analysis of co-expression of these genes within the same sample. In a subgroup of eight patients, a quantitative comparison of the relative levels of ACE gene expression was performed using a competitive polymerase chain reaction. RESULTS: Angiotensinogen and ACE expression were detected in all atrial, valve and ventricular samples examined, at levels similar to those of 'housekeeping' genes such as the transferrin receptor. Atrial renin and chymase expression were more difficult to detect, being demonstrable in only 70 and 63% of the samples, respectively. Higher levels of chymase were detected in ventricular samples than in atrial tissues. A quantitative analysis of ACE expression in eight atrial samples provided evidence of interindividual variation in the relative level of atrial ACE expression. CONCLUSIONS: The essential components of the renin-angiotensin system are co-expressed at a low level in the right atrial appendage and are detectable in other regions of the human heart. Renin and chymase genes are expressed at a lower level than the angiotensinogen and ACE genes and exhibit regional differences in expression. Interindividual variation in the relative level of ACE expression can be detected by a competitive polymerase chain reaction.

Angiotensinogen↗
PubMed1993

Brain swelling in first hour after coronary artery bypass surgery.

Six patients undergoing routine coronary artery bypass surgery were examined by magnetic resonance imaging of the brain before surgery, immediately afterwards, and 6-18 days later. Brain swelling was visible in all six patients on the immediate postoperative scan. In five patients who had later scans the swelling had subsided. No major neurological deficits were seen, and the patients were extubated successfully within 3 h of the operation. The mechanism of the cerebral swelling is uncertain, but it may provide insight into the cause of neurophysiological deficits seen after coronary artery surgery.

Aged↗
PubMed1993

Tight insertion of cytochrome b5 into large unilamellar vesicles.

Cytochrome b5 spontaneously binds to liposomes in a 'loose', or transferable form, whereas in vivo b5 binds post-translationally to the ER in the 'tight' or nontransferable form. The mechanism of tight insertion is unknown, except that it does not require SRP or energy input. The present study shows that prolonged incubation of b5 with large unilamellar vesicles (LUVs) of phosphatidylcholine results in slow conversion of the loose to the tight form, with a halftime of days. However, the process is complex. When the b5-LUVs are depleted of loose b5, by transfer of b5 to sonicated vesicles, the tight b5 is found to be concentrated to near saturating levels in a small fraction of the LUVs. If the LUVs devoid of tight b5 are recovered and then reincubated with fresh b5, the same slow transformation recurs. Apparently, a new population of vesicles, containing tight b5, is generated during the prolonged incubation with the protein. The b5-enriched LUVs contain about the same level of trapped sucrose as does the original vesicle preparation, indicating that vesicle integrity is maintained throughout the process. When fresh b5 is added to these tight b5-containing LUVs, all the freshly bound protein rapidly inserts (< 2 h) into the tight configuration. Apparently, the newly formed tight-b5/LUV vesicle population is 'insertion-active'. A model for these complex transformations is proposed.

Animals↗