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Biomedical subjects

J Lawler

Publications and source records attributed to J Lawler.

At least 127 records · Page 7Linked to original sources
PubMed1989

Effects of incomplete pulmonary gas exchange on VO2 max.

Recent evidence suggests that heavy exercise may lower the percentage of O2 bound to hemoglobin (%SaO2) by greater than or equal to 5% below resting values in some highly trained endurance athletes. We tested the hypothesis that pulmonary gas exchange limitations may restrict VO2max in highly trained athletes who exhibit exercise-induced hypoxemia. Twenty healthy male volunteers were divided into two groups according to their physical fitness status and the demonstration of exercise-induced reductions in %SaO2 less than or equal to 92%: 1) trained (T), mean VO2max = 56.5 ml.kg-1.min-1 (n = 13) and 2) highly trained (HT) with maximal exercise %SaO2 less than or equal to 92%, mean VO2max = 70.1 ml.kg-1.min-1 (n = 7). Subjects performed two incremental cycle ergometer exercise tests to determine VO2max at sea level under normoxic (21% O2) and mild hyperoxic conditions (26% O2). Mean %SaO2 during maximal exercise was significantly higher (P less than 0.05) during hyperoxia compared with normoxia in both the T group (94.1 vs. 96.1%) and the HT group (90.6 vs. 95.9%). Mean VO2max was significantly elevated (P less than 0.05) during hyperoxia compared with normoxia in the HT group (74.7 vs. 70.1 ml.kg-1.min-1). In contrast, in the T group, no mean difference (P less than 0.05) existed between treatments in VO2max (56.5 vs. 57.1 ml.kg-1.min-1). These data suggest that pulmonary gas exchange may contribute significantly to the limitation of VO2max in highly trained athletes who exhibit exercise-induced reductions in %SaO2 at sea level.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Identification of the protein 4.1 binding site to phosphatidylserine vesicles.

Previous studies have shown that protein 4.1 is a multifunctional protein that binds to spectrin, actin, glycophorins, the anion channel protein, and phosphatidylserine (PS). In this report, we have characterized the binding of protein 4.1 and its major proteolytic fragments to phospholipid vesicles. Pure 125I-labeled protein 4.1 was incubated with PS liposomes, and the free protein 4.1 was separated by ultracentrifugation. Protein 4.1 bound to PS liposomes with a high affinity. At saturation, there was 9 X 10(-3) pmol of protein 4.1 bound/pmol of PS with a Kd of 3.3 X 10(-7) M. When the protein 4.1 containing liposomes were examined in an electron microscope, the protein 4.1 was found uniformly decorating the vesicles in a rosettelike fashion. Among peripheral membrane proteins tested (spectrin, actin, ankyrin, and protein 4.1), protein 4.1 showed the highest level of binding to PS. The binding of protein 4.1 to PS, one of the principal phospholipids of the inner half of the lipid bilayer, was considerably higher than the binding to phosphatidylcholine, that is principally located in the outer half of the lipid bilayer. To identify the structural domain of protein 4.1 involved in binding to the phospholipids, a mixture of proteolytic fragments of protein 4.1 was incubated with PS liposomes. The liposomes selectively retained the 30-kilodalton (kDa) basic domain of the protein, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/isoelectric focusing.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Proteins↗
PubMed1988

Incidence of exercise induced hypoxemia in elite endurance athletes at sea level.

Recent evidence suggests that exercise-induced hypoxemia (EIH) may occur in healthy trained endurance athletes. However, at present, no data exist to describe the regularity of EIH in athletes or non-athletes. Therefore, the purpose of the present investigation was to determine the incidence of EIH during exercise in healthy subjects varying in physical fitness. Subjects (N = 68) performed an incremental cycle ergometer test to volitional fatigue with percent arterial oxyhemoglobin saturation (%SaO2) measured min-by-min. For the purpose of data analysis subjects were divided into three groups according to their level of physical training: 1) untrained (N = 16), 2) moderately trained (N = 27), and 3) elite highly trained endurance athletes (N = 25). EIH was defined as a %SaO2 of less than or equal to 91% during exercise. EIH did not occur in any of the untrained subjects or the moderately trained subjects. However, EIH occurred in 52% of the highly trained endurance athletes tested and was highly reproducible (r = 0.95; P less than 0.05). These findings further confirm the existence of EIH in healthy highly trained endurance athletes and suggests a rather high incidence of EIH in this healthy population. Hence, it is important that the clinician or physiologist performing exercise testing in elite endurance athletes recognize that EIH can and does occur in the elite endurance athlete in the absence of lung disease.

Adult↗
PubMed1988

Effects of heat shock on the expression of thrombospondin by endothelial cells in culture.

Heat-shock proteins from confluent primary cultures of bovine aortic endothelial cells were analyzed by SDS-polyacrylamide gels. In addition to the increased synthesis of the classical heat-shock proteins, there is an increase of a 180,000-mol wt polypeptide in the growth media of heat-shocked cells. Immunoprecipitation with specific antiserum indicates that the 180,000-mol wt polypeptide is thrombospondin. Assay of mRNA levels coding for thrombospondin after brief hyperthermic treatment (45 degrees C, 10 min), followed by a recovery of 2 h at 37 degrees C, results in a twofold increase in mRNA abundance. In contrast, the activation level of the 71,000-mol wt heat-shock protein mRNA occurs at an earlier time than for thrombospondin mRNA. Immunofluorescence microscopy was used to study the intracellular and extracellular distribution of thrombospondin. Thrombospondin is localized to a prominent pattern of granules of intracellular fluorescence in a perinuclear distribution in cells not exposed to heat. Upon heat treatment, the pattern of granules of intracellular fluorescence appears more pronounced, and the fluorescence appears to be clustered more about the nucleus. There are at least three pools of extracellular forms of thrombospondin: (a) the fine fibrillar extracellular matrix thrombospondin; (b) the punctate granular thrombospondin; and (c) the thrombospondin found in the conditioned medium not associated with the extracellular matrix. When bovine aortic endothelial cells are exposed to heat, the extracellular matrix staining of a fibrillar nature is noticeably decreased, with an increase in the number and degree of fluorescence of focal areas where the punctate granule thrombospondin structures are highly localized. No gross morphological changes in extracellular matrix staining of fibronectin was noted. However, the intermediate filament network was very sensitive and collapsed around the nucleus after heat shock. We conclude that the expression of thrombospondin is heat-shock stimulated.

Animals↗
PubMed1988

Cell attachment to thrombospondin: the role of ARG-GLY-ASP, calcium, and integrin receptors.

Thrombospondin is a 420,000-D glycoprotein that has recently been shown to have several properties in common with the members of a class of adhesive proteins. To characterize further the adhesive properties of thrombospondin, we have studied its ability to support cell attachment. Thrombospondin adsorbed to plastic dishes supports the attachment of human endothelial and smooth muscle cells and the monocyte-like cell line (U937) as well as normal rat kidney cells. The majority of attached cells do not spread on the solid-phase thrombospondin. The attachment of all four cell types to thrombospondin is abolished if the assay is performed in the presence of EGTA, although the cells still attach to fibronectin. If thrombospondin is adsorbed to the dishes in the presence of EGTA and then washed with buffer containing calcium before addition of the cells, attachment is still markedly inhibited, indicating that calcium affects the conformation and function of thrombospondin. Attachment of all four cell types is also markedly inhibited by the synthetic peptides gly-arg-gly-asp-ser-pro (GRG-DSP) and gly-arg-gly-asp-ala-cys (GRGDAC) but not by the control peptide gly-arg-gly-glu-ser-pro (GRG-ESP). Affinity chromatography of n-octylglucoside extracts of surface-labeled endothelial cells or smooth muscle cells on thrombospondin-Sepharose and GRG-DSP-Affigel columns was used to identify an integrin complex related to glycoprotein IIb-IIIa as an RGD-dependent receptor for thrombospondin. In addition, a monoclonal antibody (LM609) that blocks attachment of endothelial cells to vitronectin, fibrinogen, and von Willebrand factor also inhibits attachment of endothelial cells to thrombospondin. These data indicate that the attachment of cells to thrombospondin is mediated by RGD and calcium-dependent mechanisms and is consistent with the hypothesis that the GRGDAC sequence in thrombospondin is a site for interaction with an integrin receptor of the beta 3 subclass.

Amino Acid Sequence↗
PubMed1988

Linear relationship between VO2max and VO2max decrement during exposure to acute hypoxia.

The purpose of these experiments is to test the hypothesis that exercise-induced hypoxemia at sea level in highly trained athletes might be exacerbated during acute hypoxia and therefore result in correspondingly larger decrements in maximal O2 uptake (VO2max) compared with less trained individuals. Thirteen healthy male volunteers were divided into two groups according to their level of fitness: 1) trained endurance athletes (T) (n = 7), with a VO2max range of 56-75 ml.kg-1.min-1 and 2) untrained individuals (UT) (n = 6), with a VO2max range of 33-49 ml.kg-1.min-1. Subjects performed two incremental cycle ergometry tests to determine VO2max under hypoxic conditions [14% O2-86% N2, barometric pressure (PB) = 760 Torr] and normoxic conditions (21% O2-79% N2, PB = 760 Torr). Tests were single blind, randomly administered, and separated by at least 72 h. Mean percent oxyhemoglobin saturation (%SaO2) during maximal exercise under hypoxic conditions was significantly (P less than 0.05) lower in the T group (77%) compared with the UT group (86%). Furthermore, the T group exhibited larger decrements (P less than 0.05) in VO2max (normoxic-hypoxic) compared with the UT group. Finally, a significant linear correlation (r = 0.94) existed between normoxic VO2max (ml.kg-1.min-1) and delta VO2max (normoxic-hypoxic). These data suggest that highly T endurance athletes suffer more severe gas exchange impairments during acute exposure to hypoxia than UT individuals, and this may explain a portion of the observed variance in delta VO2max among individuals during acute altitude or hypoxia exposure.

Acclimatization↗
PubMed1988

Acetylated lipoproteins impair erythroid growth factor release from endothelial cells.

Endothelial cells are a known source of hematopoietic growth-enhancing factors, including platelet-derived growth factor (PDGF). In addition, endothelium interacts directly with plasma lipoproteins which have been shown to modulate hematopoiesis. To determine the relationship of these properties, we measured the release of an erythroid growth-enhancing factor from bovine endothelial cells under lipid-loaded and control conditions. Human bone marrow cells cultured under serum-free conditions form more erythroid, granulocyte/macrophage, and mixed hematopoietic colonies when supplemented with endothelial cell-conditioned medium (ECCM) than do controls (P less than 0.05). The activity is expressed over a wide range of erythropoietin, lymphocyte-conditioned medium (LCM), recombinant human interleukin-3, and colony-stimulating factor (CSF) concentrations, and is related to ECCM dose. In contrast, enhancing activity in ECCM prepared with 0-400 micrograms/ml acetylated low density lipoproteins (AcLDL) or native LDL is diminished to 0% in a dose-dependent fashion (relative to ECCM from unexposed cells or from cells incubated with very low density lipoproteins, P less than 0.05). Upon dilution, medium prepared from cells incubated with LDL shows a rightward shift in the dose-response curve for erythroid colony formation, while that prepared from AcLDL loaded cells demonstrates a downward shift, indicating that the inhibitory activities are kinetically distinct. Delipidation of ECCM prior to addition to marrow culture removes the inhibitory action of native LDL (P less than 0.05) but not that of AcLDL (P greater than 0.10). Immunochemical analysis suggests that the erythropoietic activity in ECCM is unrelated to that of PDGF, recombinant human CSF, and erythroid burst-promoting activity (BPA) present in LCM. This conclusion is supported by Northern blot analysis of endothelial cells using a cDNA probe for the v-sis homologue of the PDGF beta chain and by immunoprecipitation of metabolically labeled PDGF. The relative amounts of c-sis transcripts and of secreted PDGF were similar in endothelial cells incubated with or without AcLDL. We conclude that AcLDL impair the synthesis or release of an erythropoietic growth-enhancing factor(s) which is biologically distinct from PDGF and BPA present in LCM.

Acetylation↗
PubMed1988

Spectrin-alpha I/61: a new structural variant of alpha-spectrin in a double-heterozygous form of hereditary pyropoikilocytosis.

Recent biochemical studies have led to the identification of abnormal spectrins in the erythrocytes of patients with hereditary pyropoikilocytosis (HPP) and hereditary elliptocytosis (HE). In this report we describe the biochemical characterization of the erythrocytes from a proband with severe HPP who is doubly heterozygous for two mutant spectrins (Sp): Sp alpha I/74 and a new, previously undetected, mutant of alpha-spectrin designated Sp alpha I/61. The proband's erythrocytes are unstable when exposed to 45 degrees C, and her membrane skeletons exhibit instability to shear stress. The content of spectrin in the proband's erythrocyte membranes is decreased to 75% of control values. The amount of spectrin dimers in crude 4 degrees C spectrin extracts is increased (58%) as compared with control values (6% +/- 4%). Limited tryptic digestion reveals a marked decrease in the normal 80,000-dalton alpha I domain, an increase in the 74,000-dalton fragment that is characteristic of Sp alpha I/74, and an increase in a series of new fragments of 61,000, 55,000, 21,000, and 16,000 daltons. Both parents are asymptomatic, but they have increased amounts of spectrin dimers (17% to 25%). Limited tryptic digestion of the father's spectrin demonstrates the presence of a previously identified abnormal spectrin (Sp alpha I/74) that is characterized by a decrease in content of the 80,000-dalton peptide and an increase in concentration of the 74,000-dalton peptide. The mother's spectrin digests show a decrease in the amount of 80,000-dalton peptide and the formation of new peptides of 61,000, 55,000, 21,000, and 16,000 daltons. The data indicate that this severe form of HPP is due to the inheritance of two distinct abnormal spectrins, Sp alpha I/74 and a new spectrin mutant, Sp alpha I/61.

Anemia, Hemolytic, Congenital↗
PubMed1987

Oxygen deficit-oxygen debt relationships in ponies during submaximal treadmill exercise.

The O2 deficit-O2 debt relationship in ponies was examined at two levels of treadmill exercise. Five healthy ponies exercised for 8 min at two work rates (WR):(1) 50 m/min at 6% grade and (2) 70 m/min at 12% grade. The O2 deficit and O2 debt were calculated using open-circuit measurement of pulmonary gas exchange during the transition from rest to steady-state exercise and during recovery from exercise. The O2 deficit was significantly lower (P less than 0.05) than the O2 debt at each of the two work rates. The means (+/- SEM) for the deficit and the O2 debt, respectively, in liters were: (1) low WR = 0.45 +/- 0.12; 0.92 +/- 0.13; high WR = 1.03 +/- 0.20; 1.62 +/- 0.12. Given the observation that the O2 deficit and the O2 debt are generally equal in humans during exercise below the anaerobic threshold, these findings suggest a species difference in the O2 deficit-O2 debt relationship. It is hypothesized that the difference in O2 deficit between man and pony is related to faster VO2 kinetics for the on-transient of exercise which involves a more rapid increase in cardiac output coupled with a release of splenic stored erythrocytes.

Animals↗
PubMed1987

Clinical and laboratory study of two Caucasian families with hereditary pyropoikilocytosis and hereditary elliptocytosis.

Hereditary pyropoikilocytosis (HPP) is a severe, congenital hemolytic anemia occurring almost exclusively in black persons and characterized by extreme red blood cell anisopoikilocytosis. The authors report two unrelated white females with HPP. Both had severe hemolytic anemia at birth, red blood cell morphologic features characteristic for HPP, and increased thermal sensitivity of the red blood cells. Examination of the red blood cell membranes of both patients showed markedly unstable membrane skeletons when subjected to shear stress, spectrin dimer association defects with increased dimers, and partial spectrin deficiency. Limited tryptic digestion of the spectrin molecule from both patients yielded an abnormal pattern with a decrease in the normal 80,000-dalton alpha I domain and a concomitant increase of an abnormal 74,000-dalton peptide (Sp alpha 1/74). One parent and one sibling of one of the patients with HPP had hereditary elliptocytosis (HE) and the Sp alpha 1/74 defect. The other patient with HPP was different from others reported in that both parents were hematologically and biochemically normal. In addition, her daughter had HE and the Sp alpha 1/74 defect.

Adult↗
PubMed1987

A time-saving incremental cycle ergometer protocol to determine peak oxygen consumption.

Previously we have demonstrated that an accelerated arm ergometry testing protocol results in a higher peak oxygen consumption than continuous or discontinuous protocols reported in the literature (Brit. J. Sports Med. 20: 25-26, 1986). The purpose of this investigation was to determine if an accelerated protocol was superior to two commonly used protocols in cycle ergometry. Nine male subjects were tested on three different exercise protocols; a discontinuous test (DT), a continuous test (CT) and a new proposed "jump-max test" (JMT). The CT began at a work rate of 70 W with the power output (PO) being increased 35 W.min-1. The DT began at a work rate of 70 W; the work rate was increased by 35 W every 2 minutes with 2-minutes rest between stages. The JMT began with a 3-minute pretest to determine a PO which elicited a HR of 145 +/- 5 bpm. After a 2-minute rest, subjects began exercise at the predetermined work rate with the PO being increased 35 W.min-1. Testing sessions were terminated when subjects failed to maintain the desired PO. No significant difference (p greater than 0.05) existed in peak PO or peak oxygen consumption (VO2) between the three protocols. However, JMT protocol did result in a shorter time to exhaustion than the other protocols employed (P less than 0.05).

Adult↗
PubMed1987

Ventilatory and blood gas dynamics at onset and offset of exercise in the pony.

The purpose of these experiments was to examine the temporal pattern of arterial carbon dioxide tension (PaCO2) to assess the relationship between alveolar ventilation (VA) and CO2 return to the lung at the onset and offset of submaximal treadmill exercise. Five healthy ponies exercised for 8 min at two work rates: 50 m/min 6% grade and 70 m/min 12% grade. PaCO2 decreased (P less than 0.05) below resting values within 1 min after commencement of exercise at both work rates and reached a nadir at 90 s. PaCO2 decreased maximally by 2.5 and 3.5 Torr at the low and moderate rate, respectively. After the nadir, PaCO2 increased across time during both work rates and reached values that were not significantly different (P greater than 0.05) from rest at minute 4 of exercise. Partial pressure of O2 in arterial blood and arterial pH reflected hyperventilation during the first 3 min of exercise. At the termination of exercise PaCO2 increased (1.5 Torr) above rest (P less than 0.05), reaching a zenith at 2-3 min of recovery. These data suggest that VA and CO2 flow to the lung are not tightly matched at the onset and offset of exercise in the pony and thus challenges the traditional concept of blood gas homeostasis during muscular exercise.

Animals↗
PubMed1987

Measurement of oxygen uptake in the non-steady-state.

The purpose of these experiments was to develop and validate an open-circuit technique for measurement of gas exchange during the transition from rest to constant load steady-state exercise. The design of the open-circuit system employed to measure gas exchange in these experiments used a mixing chamber to collect the subject's expired ventilation where fractions of O2 and CO2 were determined via electronic gas analyzers. A gasometer was used to measure inspired ventilation and the analog signals from the two gas analyzers and the gasometer were sent to a microcomputer for computation of VO2. In calculating VO2, the mixed expired gas concentrations were matched with ventilatory volume using a previously determined time delay. To determine the validity of the open-circuit system, four subjects performed a series of 16 rest-to-work transitions on a cycle ergometer. In eight of the experiments, serial measurements of VO2 were obtained every 20 s for 3 min using the open-circuit mixing chamber system while the additional eight experiments used the Douglas bag technique. No significant difference (p greater than 0.05) existed between VO2 values calculated by the two techniques. Mean differences in VO2 between the two techniques during the first three 20-s measurement periods were 6, 53, and 63 ml, respectively. Using the Douglas bag technique as the standard, this represents a relative measurement error of 0.1%, 4.5%, and 3.6%, respectively, at the above time intervals. These data demonstrate that an open-circuit system employing a mixing chamber and the appropriate time delay to match expired gas fractions and ventilation is a sensitive means of measurement of VO2 in the non-steady-state.

Breath Tests↗
PubMed1987

Molecular determinants of clinical expression of hereditary elliptocytosis and pyropoikilocytosis.

The clinical severity of common hereditary elliptocytosis (HE) is highly variable, ranging from an asymptomatic carrier state to a severe hemolytic anemia. To elucidate the molecular basis of this variable clinical expression, we evaluated 56 subjects from 24 HE kindred, who carry alpha spectrin mutants characterized by a spectrin dimer (SpD) self-association defect related to a structural abnormality of the alpha I domain of spectrin. Twenty-nine subjects had common HE, 13 subjects have a closely related disorder, hereditary pyropoikilocytosis (HPP), and 14 are asymptomatic carriers. We compared the severity of hemolysis with the following biochemical parameters: (a) spectrin heterodimer self-association, as manifested by the percentage of SpD in the 4 degrees C low ionic strength spectrin extract; (b) spectrin structure, as examined by limited tryptic digestion of spectrin; and (c) spectrin content of the RBC membrane. Our analysis indicates that the severity of hemolysis may be correlated with quantitative differences in the percentage of SpD in the 4 degrees C spectrin extract, as well as the total spectrin content of the membrane. Thus, HPP subjects, who have the most severe hemolytic anemia, have the highest percentage of SpD as well as a decreased spectrin content. HE subjects and asymptomatic carriers, respectively, have a lower percentage of SpD and a normal spectrin content. Factors influencing these two determinants include functional differences between the individual spectrin mutants, the relative amounts of mutant spectrin present in the cells, the stability of mutant spectrin, and the possibility of a superimposed genetic defect involving spectrin synthesis.

Elliptocytosis, Hereditary↗
PubMed1987

Platelet antithrombin defect in malignancy: platelet protein alterations.

Sixty-eight patients with malignant disease were divided into two groups based on the results of the platelet antithrombin test (PAT). The normal group had a PAT clotting time ranging from 21.4 to 29.8 seconds, which was equivalent to 25% to 65% inactivation of the 2 U of thrombin added to the test system. The other group showed abnormal PAT clotting time, less than 21.4 seconds or less than 25% thrombin inactivation. The polypeptide composition of platelets from the two patient groups was analyzed by sodium dodecyl sulfate (SDS)-electrophoresis on 7.5% polyacrylamide gels. A polypeptide of 180,000 apparent mol wt was decreased or absent in both Coomassie blue- and Alcian blue-stained gels of the platelets from patients whose PAT was abnormal; this polypeptide comigrated with purified platelet thrombospondin. Tritium labeling of platelet surface glycoproteins by the periodate-borohydride method followed by two-dimensional electrophoresis was performed on platelets of seven patients with abnormal PAT. When they were compared with ten patients with normal PAT, a glycoprotein of 140,000 apparent mol wt with a pl of 4.5 to 5.2 was decreased in platelets of all seven patients with abnormal PAT. Nitrocellulose replicas of one-dimensional gels of platelets from 13 of 14 patients with abnormal PAT showed decreased reaction with an anti-human platelet glycocalicin antiserum. Platelets of these same patients also showed a decreased or absent platelet agglutination induced by ristocetin. Patients with normal PAT had a mean agglutination slope of 1.25 +/- 0.6 (n = 26) as compared with 0.37 +/- 0.34 (n = 26) for the abnormal PAT group (P less than .001). Results indicate that platelets from a subpopulation of tumor patients characterized by decreased platelet antithrombin activity have alterations in two platelet glycoproteins, identified as GPIb and thrombospondin.

Antithrombins↗