Biomedical subjects

J Lawler

Publications and source records attributed to J Lawler.

At least 109 records · Page 6Linked to original sources

PubMed1992

Endurance training-induced increases in expiratory muscle oxidative capacity.

Recent evidence demonstrates that endurance exercise training improves the oxidative capacity of the major mammalian inspiratory muscle (e.g., costal diaphragm). In contrast, no data exist concerning the effects of exercise training on abdominal expiratory muscles. We tested the hypothesis that 12 wk of endurance exercise training would significantly increase the activity of selected beta oxidation and Krebs cycle enzymes of abdominal expiratory muscles of the rat. To test this hypothesis two groups of female Sprague-Dawley rats were studied: group 1, continuous exercise training (n = 13); and group 2, sedentary control (n = 6). Exercise trained animals ran 5 d.wk-1 on a motorized treadmill for 45 min.d-1 at approximately 75-80% VO2max. When compared with controls, exercise training resulted in elevated (P less than 0.05) activities of 3-hydroxy-acyl-Co-A dehydrogenase (HADH) and citrate synthase (CS) in two abdominal expiratory muscles (rectus abdominus and external obliques). In contrast, training did not alter (P greater than 0.05) CS or HADH activity in the internal obliques/transversus abdominus muscles. In general, the training-induced increases in expiratory muscles CS activity were relatively small (approximately 10-13%) when compared with the training-induced increase in CS activity in the plantaris muscle (approximately 44%). These data demonstrate that continuous exercise training results in small but statistically significant improvements in the oxidative and beta oxidation capacities of expiratory muscles.

3-Hydroxyacyl CoA Dehydrogenases↗

PubMed1991

Cloning and sequencing of chicken thrombospondin.

Thrombospondin is a multifunction adhesive protein with the ability to bind proteoglycans, cell surface receptors, other proteins, and calcium ions. Several sequence motifs for some of these interactions have been identified in human thrombospondin. To evaluate the potential functional significance of these sequences and to begin a study of the evolution of thrombospondin, we have isolated and sequenced thrombospondin cDNA clones from a chicken embryo library. Comparison of the chicken and human sequences reveals that the NH2-terminal heparin-binding domains are only 34% identical. By contrast, the type 3 repeats and the COOH-terminal domains are 80 and 82% identical, respectively, when comparing human and chicken sequences. Potential cell recognition sequences of RGD and VTCG are conserved, with the chicken sequence containing an additional copy of the VTCG sequence. Whereas substitutions occur in the two potential heparin-binding motifs that have human counterparts, the chicken sequence contains a third potential heparin-binding motif. The results indicate that the evolutionary constraints on the various types of cell-binding motifs may be quite different.

Amino Acid Sequence↗

PubMed1991

Age-related changes in enzyme activity in the rat diaphragm.

Limited data exist concerning the effect of growth and aging on the metabolic properties of the diaphragm. Therefore, we investigated age-related changes in protein concentration and glycolytic and Krebs cycle enzyme activity in the diaphragm as well as the plantaris muscle of female Sprague-Dawley rats ranging in age from 1 to 12 months. Samples from the costal and crural diaphragm and the plantaris muscle were obtained from 38 animals in the following age groups: (1) 1 month old (N = 7); (2) 4 month old (N = 6) (3) 6 month old (N = 13); and (4) 12-month-old (N = 12). Body weight and diaphragm weight increased rapidly by a factor of 6 and in parallel during 1-4 months postpartum before reaching a plateau at 6 months of age. No significant difference (P greater than 0.05) existed in the ratio of diaphragm weight to body weight among age groups. Protein concentration was significantly higher (P less than 0.05) in the costal diaphragm and plantaris at 4 and 6 months when compared to 1 and 12 months of age. In the crural diaphragm, protein concentration was significantly lower (P less than 0.05) at 1 month postpartum when compared to all other age groups. Succinate dehydrogenase (SDH) activity was significantly higher (P less than 0.05) at 1 month of age in the plantaris, the costal diaphragm and the crural diaphragm when compared to older animals. In contrast, the activity of lactate dehydrogenase (LDH) in the plantaris, the costal diaphragm and the crural diaphragm was significantly lower (P less than 0.05) in the 1-month-old animals when compared to all other ages.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

PubMed1991

Characterization of the murine thrombospondin gene.

Thrombospondin is an adhesive glycoprotein that supports cell attachment, spreading, and migration. The murine thrombospondin gene is approximately 18 kb in length and includes 22 exons. Interspecific backcross analysis using progeny derived from matings of (C57BL/6J x Mus spretus) F1 x C57BL/6J mice indicates that the thrombospondin gene is tightly linked to the Fshb, Actcl, Ltk, and B2M loci on murine chromosome 2. The sequence of the murine gene is very similar to that of the human gene in (1) regions of the promoter, (2) the coding region, and (3) the 3'-untranslated region. The predicted amino acid sequence of the mature murine thrombospondin subunit is 95.1% identical to that of the human. The sequences of these two species are most similar at the regions containing the type 1, 2, and 3 repeats as well as the COOH-terminal globular domain. The thrombospondin promoter is similar to the 5' flanking region of some housekeeping and growth control genes in that it contains multiple GC-rich regions and lacks a CAAT box. The presence of various consensus sequences suggests that thrombospondin gene expression is regulated by cAMP, cytokines, and steroid hormones.

Amino Acid Sequence↗

PubMed1991

Evidence for an alveolar-arterial PO2 gradient threshold during incremental exercise.

Examination of the alveolar-to-arterial O2 tension (A-a PO2 difference) provides a method of examining the efficiency of pulmonary gas exchange during exercise. At present, considerable confusion exists as to the exact pattern of the A-a PO2 difference during incremental exercise. We tested the hypothesis that the A-a PO2 difference during incremental exercise is alinear with respect to metabolic rate. Measurements of the A-a PO2 difference were made on six healthy male subjects during incremental exercise under sea level conditions (PIO2 = 149 torr). An alinear model best described the relationship between the A-a PO2 difference and metabolic rate; only small increases in the A-a PO2 difference occurred at low work rates followed by a rapid increase at higher work rates. The existence of a "A-a PO2 difference threshold" was mathematically confirmed by the use of a computer algorithm to define inflection points. These data provide evidence that the relationship between the A-a PO2 difference and metabolic rate is alinear and that a metabolic threshold exists for a rapid increase in the A-a PO2 difference. We conclude that the efficiency of pulmonary gas exchange during exercise is unaltered from rest during low-to-moderate power outputs, however, high intensity exercise compromises pulmonary gas exchange efficiency as evidenced by a significant widening of the A-a PO2 difference.

Adult↗

PubMed1991

Influence of a carbohydrate-electrolyte beverage on performance and blood homeostasis during recovery from football.

This study compared the efficacy of a 7% glucose polymer beverage containing electrolytes (GP) versus a nonnutrient, nonelectrolyte placebo (P) in maintaining blood homeostasis during recovery from football and determined whether consumption of the GP beverage improved anaerobic performance immediately after football competition when compared with the placebo. Forty-four high school football players participated in a 50-play scrimmage designed to simulate game conditions. At each of six periods before and during the scrimmage, players consumed 170 ml of the GP or P beverage. Eight maximal-effort 40-yd sprints (40-sec rest intervals) were performed before and after the scrimmage to assess the decrement in anaerobic performance from the scrimmage. Venous blood samples were drawn before and after the scrimmage and analyzed. The pre- to postscrimmage differences in mean and peak sprint velocities did not differ between treatments, nor did body weight and plasma. In contrast, the percent decrease in plasma volume was significantly greater in the P group. Postscrimmage increases in glucose and insulin were greater in the GP group. These data suggest that CHO-electrolyte drinks do not prevent a decline in anaerobic performance when compared to water, but a CHO-electrolyte drink is more effective in maintaining PV than water during recovery from anaerobic exercise.

Adolescent↗

PubMed1991

Physiological correlates to 800 meter running performance.

Much of the previous research efforts aimed at determining those physiological characteristics that contribute to distance running success have centered around distances greater than 1500 meters with little attention to events such as the 800 meter run. Therefore, this investigation examined the relationship between selected physiological and body composition, characteristics and performance in an 800 meter run. Measurements of body composition, VO2max, running economy, and performance times for 100 and 300 meter dashes were obtained on 11 male track athletes. Stepwise multiple regression analysis was performed using 800 meter race time as the dependent variable. Although the combination of 300 and 100 meter run times, percent body fat, running economy and VO2 max as independent variables accounted for the greatest amount of total variance (r2 = .89), the additional variance explained by the model did not increase significantly (p greater than 0.05), when VO2max, percent body fat, and running economy were added to a model which contained 300 and 100 meter run time (r2 = .85) as the explanatory variables. These data offer additional support for the notion that much of the intramuscular ATP produce and utilized during an 800 meter run comes from anaerobic metabolic pathway.

Adult↗

PubMed1990

Structural and functional heterogeneity of alpha spectrin mutations involving the spectrin heterodimer self-association site: relationships to hematologic expression of homozygous hereditary elliptocytosis and hereditary pyropoikilocytosis.

Defects involving alpha spectrin (Sp) are found in patients with hereditary elliptocytosis and a related disorder, hereditary pyropoikilocytosis (HPP). We have previously found that the severity of hemolysis was related to the total spectrin content of the cells and the percentage of unassembled dimeric Sp (SpD) in the membranes, which, in turn, reflected the amount of mutant Sp in the cell. However, no data are available comparing differences in the function of various alpha Sp mutations to clinical severity. We now report studies of nine homozygotes or double heterozygotes for four alpha Sp mutations: alpha 1/74, alpha 1/46, alpha 1/65, and alpha 1/61, whose red blood cells (RBCs) contained only the mutant Sp and no normal Sp. Sp alpha 1/74, Sp alpha 1/46, and alpha 1/65 homozygotes differed strikingly in the severity of hemolysis that correlated with the severity of mutant Sp dysfunction, as reflected by the fraction of unassembled SpD in the membranes and the self-association of mutant Sp on inside-out vesicles. Homozygotes for Sp alpha 1/74 had a very severe hemolytic anemia and their SpD were virtually incapable of self-association, whereas SpD alpha 1/46 were not as severely affected. The Sp alpha 1/65 homozygotes had a relatively mild hemolytic anemia and their SpD showed the least impairment of function. Ultrastructural examination of membrane skeletons from subjects whose SpD self-association was severely impaired showed gross skeletal disruption and loss of hexagonal structure. In striking contrast, the homozygote for the mildly dysfunctional Sp alpha 1/65 had only a moderate disruption of the skeleton. Some of the homozygous or doubly heterozygous subjects also exhibited a partial deficiency of Sp that correlated with a RBC morphology characteristic of HPP, namely, marked microspherocytosis with virtual absence of elliptocytes. These data demonstrate striking differences in the function and structure of various alpha Sp mutants that underlie differences in clinical expression.

Adolescent↗

PubMed1990

Fluid replacement drinks during high intensity exercise: effects on minimizing exercise-induced disturbances in homeostasis.

The purpose of these experiments was to examine the influence of various fluid replacement drinks on exercise-induced disturbances in homeostasis during heavy exercise. Nine trained cyclists performed constant load exercise on a cycle ergometer to fatigue on three occasions with 1-week separating experiments. The work rate was set initially at approximately 85% of VO2max (range 82-88%) with fatigue being defined as a 10% decline in power output below the initial value. During each experiment subjects consumed one of the following three beverages prior to and every 15 min during exercise: (1) non-electrolyte placebo (NEP; 31 mosmol.kg-1); (2) glucose polymer drink containing electrolytes (GP; 7% CHO, 231 mosmol.kg-1), and (3) electrolyte placebo drink without carbohydrate (EP; 48 mosmol.kg-1). Both the GP and EP beverage contained sodium citrate/citric acid (C) as a flavoring agent while C was not contained in the NEP drink. Although seven of nine subjects worked longer during the GP and EP treatment when compared with the NEP trial, the difference was not significant (P greater than 0.05). No differences (P greater than 0.05) existed between the GP and EP treatments in performance time. Exercise changes in rectal temperature, heart rate, delta % plasma volume and plasma concentrations of total protein, free fatty acids, glucose, lactate, potassium, chloride, calcium, and sodium did not differ (P greater than 0.05) between trials.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1990

Complete amino acid sequence and homologies of human erythrocyte membrane protein band 4.2.

The complete amino acid sequence for human erythrocyte band 4.2 has been derived from the nucleotide sequence of a full-length 2.35-kilobase (kb) cDNA. The 2.35-kb cDNA was isolated from a human reticulocyte cDNA library made in the expression vector lambda gt11. Of the 2348 base pairs (bp), 2073 bp encode 691 amino acids representing 76.9 kDa (the SDS/PAGE molecular mass is 72 kDa). RNA blot analysis of human reticulocyte total RNA gives a message size for band 4.2 of 2.4 kb. The amino acid sequence of band 4.2 has homology with two closely related Ca2(+)-dependent cross-linking proteins, guinea pig liver transglutaminase (protein-glutamine gamma-glutamyltransferase; protein-glutamine: amine gamma-glutamyltransferase, EC 2.3.2.13) (32% identity in a 446-amino acid overlap) and the a subunit of human coagulation factor XIII (27% identity in a 639-amino acid overlap), a transglutaminase that forms intermolecular gamma-glutamyl-epsilon-lysine bonds between fibrin molecules. The region of greatest identity includes a 49-amino acid stretch of band 4.2, which is 69% and 51% identical with guinea pig liver transglutaminase and the a subunit of factor XIII, respectively, within the regions that contain the active sites of these enzymes. Significantly, within the five contiguous consensus residues of the transglutaminase active site, Gly-Gln-Cys-Trp-Val, band 4.2 has an alanine substituted for cysteine (which is apparently essential for activity). Consistent with this active site substitution, erythrocyte membranes or inside-out vesicles, which contain band 4.2, show no evidence of transglutaminase activity by two types of in vitro assay.

Amino Acid Sequence↗

PubMed1990

cDNA sequence for human erythrocyte ankyrin.

The cDNA for human erythrocyte ankyrin has been isolated from a series of overlapping clones obtained from a reticulocyte cDNA library. The composite cDNA sequence has a large open reading frame of 5636 base pairs (bp) with the complete coding sequence for a polypeptide of 1879 amino acids with a predicted molecular mass of 206 kDa. The derived amino acid sequence contained 194 residues that were identical to those obtained by direct amino acid sequencing of 11 ankyrin proteolytic peptides. The primary sequence contained 23 highly homologous repeat units of 33 amino acids within the 90-kDa band 3 binding domain. Two cDNA clones showed evidence of apparent mRNA processing, resulting in the deletions of 486 bp and 135 bp, respectively. The 486-bp deletion resulted in the removal of a 16-kDa highly acidic peptide, and the smaller deletion had the effect of altering the COOH terminus of the molecule. Radiolabeled ankyrin cDNAs recognized two erythroid message sizes by RNA blot analysis, one of which was predominantly associated with early erythroid cell types. An ankyrin message was also observed in RNA from the human cerebellum by the same method. The ankyrin gene is assigned to chromosome 8 using genomic DNA from a panel of sorted human chromosomes.

Amino Acid Sequence↗

PubMed1990

Endurance-training-induced cellular adaptations in respiratory muscles.

Controversy exists concerning the adaptability of mammalian respiratory muscles in response to endurance training. We examined the effects of 8 wk of progressive treadmill exercise (45 min/day 5 days/wk) on the biochemical adaptations of rat diaphragm and intercostal muscles. Female Sprague-Dawley rats were randomly assigned to a sedentary control (n = 10) or an exercise-training group (n = 10). Endurance training resulted in an enhanced oxidative capacity in the anterior costal diaphragm as evidenced by a 29% increase (P less than 0.05) in the activity of succinate dehydrogenase (SDH) in trained animals compared with controls (4.15 +/- 0.13 vs. 3.21 +/- 0.17 mumol.g-1.min-1). Similarly, SDH activity in the intercostal muscles was 32% greater (P less than 0.05) in the trained animals than in the untrained animals (1.72 +/- 0.11 vs. 1.30 +/- 0.06 mumol.g-1.min-1). In contrast, the crural region of the diaphragm showed no significant increase (P greater than 0.05) in oxidative capacity as a result of the training program (3.28 +/- 0.12 vs. 3.13 +/- 0.18). Furthermore, training did not alter (P less than 0.05) lactate dehydrogenase activity in the intercostals or in the crural or the costal diaphragm. These data demonstrate that the oxidative capacity of the costal diaphragm and the intercostal muscles can be enhanced by increasing respiratory loads via regular endurance exercise. We speculate that the lack of metabolic adaptation in the crural region of the diaphragm was not due to limited plasticity of the fibers in this area but to failure to the exercise-training program to provide the appropriate stimulus for cellular adaptation.

Adaptation, Physiological↗

PubMed1990

Regional metabolic differences in the rat diaphragm.

This study characterized the biochemical properties of the rat diaphragm by measuring the activities of selected citric acid cycle and glycolytic enzymes. The diaphragm was removed from 10 female Sprague-Dawley rats (180 days old) and dissected into five discrete anatomic regions: crural (region 1), left posterior costal (region 2), left anterior costal (region 3), right anterior costal (region 4), and right posterior costal (region 5). Sections were assayed for total protein concentration and the activities of succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH). The SDH activity in the crural region was approximately 18% lower (P less than 0.05) than that in any costal region. Furthermore, protein concentration was significantly lower (P less than 0.05) in the crural region compared with all costal regions. In contrast, costal regions 2-5 did not significantly differ from each other in protein concentration or SDH activity. LDH activity did not differ significantly (P greater than 0.05) between regions. Finally, the LDH-to-SDH activity ratio was significantly higher (P less than 0.05) in the crural diaphragm compared with all costal regions. We conclude that the crural region of the rat diaphragm is significantly lower in oxidative capacity than all the costal regions. Investigators who use a rodent model to study diaphragmatic function and plasticity should consider the oxidative heterogeneity of the diaphragm when designing experiments.

Animals↗

PubMed1990

Effects of thrombospondin antibody on the recovery of endothelial cells from hyperthermia.

In addition to the increased synthesis of the classical heat-shock proteins (28,000, 71,000, 73,000, 90,000 and 100,000 Mr polypeptides) there is also an increase of thrombospondin in the growth medium of endothelial cells exposed to hyperthermia. The effect of a monoclonal antibody to thrombospondin on the recovery of endothelial cells from hyperthermia as it relates to cytoskeletal organization and cell spreading was assessed. The antibody interacts with the heparin-binding domain of thrombospondin in the extracellular matrix of cells. We report that during recovery from thermal insult at 37 degrees C, intermediate filaments, stress fibres and microtubules show distinct time-recovery characteristics in bovine aortic endothelial cells; that in the presence of this antibody the cytoskeleton is notably altered; that this antibody causes retraction of endothelial cell processes; and that the recovery of the cytoskeleton in endothelial cells exposed to hyperthermia is prevented by the thrombospondin antibody in the time frame examined. Our data suggest that the recovery of cells from heat shock requires the integrity of thrombospondin and its interactions.

Actins↗

PubMed1989

An integrin receptor on normal and thrombasthenic platelets that binds thrombospondin.

The members of the integrin family of membrane glycoprotein heterodimer complexes function as cell surface receptors for adhesive proteins. We report here on the identification of two integrins on the surface of human platelets that bind to thrombospondin. When platelet membrane proteins are radiolabeled with 125I-lactoperoxidase, solubilized in n-octylglucoside, (Boehringer Mannheim Biochemicals, Indianapolis, IN), and applied to a column of thrombospondin-Sepharose, both complexes are bound to the column and specifically eluted with the peptide GRGDSP. One of these integrins, glycoprotein (GP) IIb-IIIa, appears to bind relatively weakly. The second integrin shares the same beta subunit (beta 3 or GPIIIa), but has a distinct alpha subunit that comigrates with the alpha subunit (alpha v) of the vitronectin receptor (VnR) on endothelial cells and reacts with a monoclonal antibody, LM142, which was raised against an integrin from M21 melanoma cells. The alpha v beta 3 integrin is present on a variety of cell types and appears to act as a receptor for thrombospondin on endothelial and smooth muscle cells. On endothelial and M21 melanoma cells this receptor is also involved in adhesion to fibrinogen, vitronectin, and von Willebrand factor (vWF). The alpha v beta 3 integrin is present at approximately equal levels on normal and thrombasthenic platelets, whereas levels of GPIIb-IIIa are greatly reduced on thrombasthenic platelets. The alpha v beta 3 integrin on thrombasthenic platelets also binds to thrombospondin-Sepharose and can be eluted with the peptide GRGDSP. These data indicate that the alpha v beta 3 integrin on platelets, endothelial cells, and smooth muscle cells functions as an Arg-Gly-Asp (RGD)-dependent receptor for thrombospondin.

Blood Platelet Disorders↗

PubMed1989

CD36 directly mediates cytoadherence of Plasmodium falciparum parasitized erythrocytes.

Erythrocytes infected with P. falciparum express knob-like adhesion structures that allow the infected cells to cling to the postcapilliary endothelium of characteristic host organs. At present, the mechanism of cytoadherence is not fully understood. While parasitized erythrocytes have been shown to specifically bind to the platelet/matrix molecule thrombospondin, adherence to suitable target cells can also be blocked by monoclonal antibody OKM5, which recognizes a surface molecule expressed by hematopoietic cells and endothelium. In apparent reconciliation of these findings, it has been reported that the OKM5 antigen (CD36) is a receptor for thrombospondin. Here we report that expression of a CD36 cDNA clone in COS cells supports cytoadherence of parasitized erythrocytes but does not support increased binding of purified human thrombospondin.

Amino Acid Sequence↗

PubMed1989

Severe hemolysis and red cell fragmentation caused by the combination of a spectrin mutation with a thrombotic microangiopathy.

Two patients are described who presented with severe hemolysis and erythrocyte fragmentation. One patient had renal allograft rejection and disseminated intravascular coagulation, and the other had thrombotic thrombocytopenia purpura. The severity of hemolysis and the red cell abnormalities were considerably more profound than usually seen in patients with thrombotic microangiopathies. After evaluation of blood smears prepared before the onset of the disease and biochemical characterization of proteins of the red blood cell skeleton, a mutation of the skeletal protein spectrin, designated Sp alpha l/65, was identified. In the heterozygous form, this mutation manifests as mild, often asymptomatic, hereditary elliptocytosis. We conclude that in these two patients with thrombotic microangiopathy, the intrinsic red cell membrane instability resulting from the underlying skeletal defect aggravated the mechanical red cell fragmentation, producing morphological features similar to the severe hemolytic form of hereditary elliptocytosis or hereditary pyropoikilocytosis.

Adolescent↗

PubMed1989

Exercise performance following intense, short-term ventilatory work.

Evidence exists to indicate that prolonged ventilatory work fatigues respiratory muscles and may limit exercise tolerance. However, the effects of short-duration, high-intensity ventilatory work on subsequent exercise remains in question. We tested the hypothesis that intense short-term volitional hyperpnea would result in respiratory muscle fatigue and would therefore hinder subsequent exercise tolerance. Pulmonary function was determined in ten healthy, male subjects before and after two constant load exercise tests to exhaustion on a cycle ergometer. Test 1 was a preliminary test to determine VO2max, peak exercise VE, and peak exercise power output. Test 2 was a constant load (85% peak power output) exercise test to exhaustion. Test 3 was identical to test 2 but was preceded by 10 min of volitional, isocapnic hyperpnea (85% of peak exercise V.E) at a controlled frequency and tidal volume. Pulmonary function measures (FVC, FEV1, FEV1/FVC, and peak flow) were not significantly (P less than 0.05) altered by the volitional hyperpnea. Ventilation and gas exchange variables (VO2, VE, f, end-tidal PO2 and PCO2, VE/VO2, VE/VCO2, %SaO2) during exercise and time to exhaustion were not significantly (P less than 0.05) different between treatments. These experiments failed to show any effect of short-term ventilatory work on pulmonary function or subsequent exercise performance.

Adult↗