Comparison and reliability of two non-invasive acetylene uptake techniques for the measurement of cardiac output.

Comparison and reliability of two non-invasive acetylene uptake techniques for the measurement of cardiac output. Thirteen trained male cyclists performed CO2 rebreathing (CO2RB) at intensities from rest to 200 W, and open-circuit acetylene uptake (OpCirc) and single-breath acetylene uptake (SB) at intensities from rest to 300 W, with all procedures using 50 W increments. Oxygen consumption VO2 cardiac output Q and heart rate (HR), were measured at each stage, and the values for each variable were compared within each intensity to determine reliability of the measuring device. Both the OpCirc and SBs were shown to be reliable measures of cardiac output (r = 0.95 and 0.92, respectively) with decreasing coefficients of variation (CV) as intensity increased, and were similar to published data. The Q-VO2 relationship using the SB diverged from the regression line for OpCirc and CO2RB. Linear regression of the Q--VO2 relationship for CO2RB was y = 6.18 x VO2 + 2.59 for OpCirc was y = 6.12 x VO2 + 2.98 and for SB was y = 5.05 x VO2 + 3.76. The OpCirc and SBs were both shown to be reliable techniques for measuring cardiac output, comparable to previously reported cardiac output measurements, and suitable for use in exercise testing. However, the SB, requiring a constant, slow exhalation rate, made the procedure difficult to perform at higher exercise intensities.

Evaluation of the Monark Wingate ergometer by direct measurement of resistance and velocity.

The purpose of this study was to assess the accuracy of the new basket-loaded Wingate ergometer introduced by Monark (Model 834E). Velocity was measured directly from the pedal switch while tension was measured with transducers on each end of the brake lacing. Moment of inertia of the flywheel was determined and accounted for in the calculation of power. Constant load tests (39.24 to 98.1 N), were done at pedaling speeds from 80 to 140 r x min(-1) (flywheel angular velocity = 30-50 rad x s(-1)). The load transmitted to the lacing at the front and back of the flywheel was 95.5 +/- 0.8% (mean +/- SEM) and 6.71 +/- 0.8%, respectively, of the load in the basket. Thus, the resultant tension (front minus back) was on average 88.8 +/- 0.57% of the applied load. The velocity recorded by the Monark Wingate Ergometer computer program (MWECP) was the same (100.4 +/- 1.56%) as that determined from the pedal switch directly. Five male mountain bikers performed a 30-s all-out test. Peak power calculated by MWECP (1181 +/- 55W) was always higher (p < .01) than that calculated from direct measures of tension and velocity (1102 +/- 66W), when not taking into account the moment of inertia. These experiments suggest that the basket-loaded Monark Wingate ergometer does not provide a correct calculation of power because of incomplete load transmission to the flywheel.

Changes in glutamine and glutamate concentrations for tracking training tolerance.

PURPOSE: The purpose was to monitor high-performance athletes throughout training macrocycles and competitions and examine the changes in plasma glutamine (Gm) and glutamate (Ga) concentrations in order to develop a model of tolerance to training. METHODS: Plasma glutamine and glutamate concentrations of 52 National team athletes (31 male and 21 female) divided into male and female groups of speed skating, swimming, and cross-country skiing were measured in an early season rested condition to determine highest Gm and lowest Ga concentrations and over 2-4 macrocycles, which included heavy training to establish lowest Gm and highest Ga concentrations. RESULTS: In the rested condition, there were no differences within and between the male and female groups, excluding five athletes (OTA) who became overtrained in heavy training. The mean (+/-SD) Gm concentration was 585 +/- 54 micromol x L(-1), Ga concentration 101 +/- 16 micromol x L(-1), and Gm/Ga ratio 5.88 +/- 0.84 micromol x L(-1). The OTA had a significantly higher Ga concentration of 128 +/- 16 micromol x L(-1) and lower Gm/Ga ratio of 4.43 +/- 0.49 micromol x L(-1) than all the other groups. In heavy training, there was a significant decrease (P < 0.05) in Gm concentration to 522 +/- 53 micromol x L(-1), significant increase in Ga concentration to 128 +/- 19 micromol x L(-1) and significant decrease in Gm/Ga ratio to 4.15 +/- 0.57 micromol x L(-1). The OTA Gm concentration of 488 +/- 31 micromol x L(-1) was significant lower than only the male speed skating and swimming groups. However, the Ga concentration of 171 +/- 17 micromol x L(-1) and Gm/Ga ratio of 2.88 +/- 0.27 micromol x L(-1) were significantly higher and lower respectively than all other groups. CONCLUSIONS: Based on the changes in Gm and Ga concentration under different training conditions, we propose an athlete tolerance to training model where glutamine concentration reflects tolerance to volume of work and glutamate concentration reflects tolerance to high intensity training. We suggest that the Gm/Ga ratio may globally represent overall tolerance to training.

Identification of ascorbic acid-deficient Arabidopsis thaliana mutants.

Vitamin C (l-ascorbic acid) is a potent antioxidant and cellular reductant present at millimolar concentrations in plants. This small molecule has roles in the reduction of prosthetic metal ions, cell wall expansion, cell division, and in the detoxification of reactive oxygen generated by photosynthesis and adverse environmental conditions. However, unlike in animals, the biosynthesis of ascorbic acid (AsA) in plants is only beginning to be unraveled. The previously described AsA-deficient Arabidopsis mutant vtc1 (vitamin c-1) was recently shown to have a defect in GDP-mannose pyrophosphorylase, providing strong evidence for the recently proposed role of GDP-mannose in AsA biosynthesis. To genetically define other AsA biosynthetic loci, we have used a novel AsA assay to isolate four vtc mutants that define three additional VTC loci. We have also isolated a second mutant allele of VTC1. The four loci represented by the vtc mutant collection have been genetically characterized and mapped onto the Arabidopsis genome. The vtc mutants have differing ozone sensitivities. In addition, two of the mutants, vtc2-1 and vtc2-2, have unusually low levels of AsA in the leaf tissue of mature plants.

Flexor tendon suture methods: a quantitative analysis of suture material within the repair site.

This study compared the cross-sectional area and volume occupied by suture material at the repair site in three common methods of flexor tendon repair. A total of 51 human cadaveric tendons were studied. Zone II flexor digitorum profundus tendon lacerations were created and then repaired using the techniques described by Kessler, Tajima, and Savage. Quantitative cross-sectional area and volumetric measurements of suture material within each repair site were determined using a digital image analysis system. The Tajima repair occupied 27% of the tendon area at the repair site, while the Savage and Kessler repairs occupied 18% and 2%, respectively.

Genetic evidence for the role of GDP-mannose in plant ascorbic acid (vitamin C) biosynthesis.

Vitamin C (L-ascorbic acid; AsA) acts as a potent antioxidant and cellular reductant in plants and animals. AsA has long been known to have many critical physiological roles in plants, yet its biosynthesis is only currently being defined. A pathway for AsA biosynthesis that features GDP-mannose and L-galactose has recently been proposed for plants. We have isolated a collection of AsA-deficient mutants of Arabidopsis thaliana that are valuable tools for testing of an AsA biosynthetic pathway. The best-characterized of these mutants (vtc1) contains approximately 25% of wild-type AsA and is defective in AsA biosynthesis. By using a combination of biochemical, molecular, and genetic techniques, we have demonstrated that the VTC1 locus encodes a GDP-mannose pyrophosphorylase (mannose-1-P guanyltransferase). This enzyme provides GDP-mannose, which is used for cell wall carbohydrate biosynthesis and protein glycosylation as well as for AsA biosynthesis. In addition to genetically defining the first locus involved in AsA biosynthesis, this work highlights the power of using traditional mutagenesis techniques coupled with the Arabidopsis Genome Initiative to rapidly clone physiologically important genes.

Complementation of the Arabidopsis pds1 mutation with the gene encoding p-hydroxyphenylpyruvate dioxygenase.

Plastoquinone and tocopherols are the two major quinone compounds in higher plant chloroplasts and are synthesized by a common pathway. In previous studies we characterized two loci in Arabidopsis defining key steps of this biosynthetic pathway. Mutation of the PDS1 locus disrupts the activity of p-hydroxyphenylpyruvate dioxygenase (HPPDase), the first committed step in the synthesis of both plastoquinone and tocopherols in plants. Although plants homozygous for the pds1 mutation could be rescued by growth in the presence of homogentisic acid, the product of HPPDase, we were unable to determine if the mutation directly or indirectly disrupted HPPDase activity. This paper reports the isolation of a cDNA, pHPPD, encoding Arabidopsis HPPDase and its functional characterization by expression in both plants and Escherichia coli. pHPPD encodes a 50-kD polypeptide with homology to previously identified HPPDases, including 37 highly conserved amino acid residues clustered in the carboxyl region of the protein. Expression of pHPPD in E. coli catalyzes the accumulation of homogentisic acid, indicating that it encodes a functional HPPDase enzyme. Mapping of pHPPD and co-segregation analysis of the pds1 mutation and the HPPD gene indicate tight linkage. Constitutive expression of pHPPD in a pds1 mutant background complements this mutation. Finally, comparison of the HPPD genomic sequences from wild type and pds1 identified a 17-bp deletion in the pds1 allele that results in deletion of the carboxyterminal 26 amino acids of the HPPDase protein. Together, these data conclusively demonstrate that pds1 is a mutation in the HPPDase structural gene.

Effects of short-term altitude training and tapering on left ventricular morphology in elite swimmers.

Short or long-term athletic training has been associated with left ventricular (LV) morphological adaptations, including increases in wall thickness, cavity dimension and estimated LV mass. A limitation of previous studies assessing the 'athlete heart' was that exercise training was performed at sea level. Since the 1968 Olympic summer games a popular method of maximizing athletic performance has been to use altitude training (AT) as a means of improving sea level performance. However, the effects of short term AT and taper training on LV morphology have not been well studied. Based on this limitation, the effects of three weeks of intense AT (1848 m) or low level control training (CT) (1050 m) followed by two weeks of taper training were investigated in 15 elite swimmers between 16 and 21 years of age. Short term AT or CT training followed by two weeks of taper training was not associated with alterations in LV diastolic cavity dimension (AT pre 53.3 +/- 2.8 mm versus post 52.6 +/- 4.3 mm; CT pre 52.9 +/- 3.7 mm versus post 51.2 +/- 4.0 mm), ventricular septal wall thickness (AT pre 9.6 +/- 1.0 mm versus post 9.4 +/- 1.1 mm; CT pre 8.4 +/- 1.2 mm versus post 8.6 +/- 1.1 mm), estimated LV mass (AT pre 186.4 +/- 45.8 g versus post 190.0 +/- 48.2 g; CT pre 159.1 +/- 35.8 g versus post 160.1 +/- 40.8 g) or fractional shortening (AT pre 36.8 +/- 3.5% versus post 34.8 +/- 2.7%; CT pre 32.6 +/- 5.0% versus post 32.8 +/- 4.7%). However, a main time effect, independent of training intervention, was observed for posterior wall thickness (pre 8.7 +/- 1.4 mm versus post 9.3 +/- 1.1 mm, P < 0.05). Therefore, with the exception of posterior wall thickness, short term AT followed by two weeks of taper training appears not to be associated with alterations in LV morphology or systolic function.

Effects of endurance training on transient oxygen uptake responses in cyclists.

The aim of this study was to determine the alterations in oxygen uptake kinetics following endurance training in previously trained athletes. Sixteen competitive cyclists completed 8 weeks of supervised endurance cycle training. Ventilatory threshold, maximal oxygen uptake (VO2max), oxygen uptake kinetics and simulated 40-km time-trial tests were performed three times over a 4-week period before training, and then after 4 and 8 weeks of training. The protocol for measuring oxygen uptake kinetics consisted of three square-wave increments from unloaded cycling to a power output of 78 W followed by a single increment from 78 to 156 W. No significant differences in any variables were observed over the pre-training period. The ventilatory threshold and VO2max increased, and the time for 40 km decreased (P < 0.05) with training. Shorter VO2 time constants and lower heart rates were observed during the protocol for measuring oxygen uptake kinetics (same absolute power output) post-training. These results indicate that oxygen uptake kinetics may be improved with endurance training in previously trained athletes.

The effect of salbutamol on performance in endurance cyclists.

The effect of salbutamol (S) on cycling performance was examined in 15 highly trained non-asthmatic male cyclists. A double-blind, randomized cross-over design was used with S or placebo (P) administered using a metered-dose inhaler and a spacer device 20 min before each testing session. The S dose was 400 micrograms (four puffs), which is twice the normal therapeutic level. Subjects were habituated to all the laboratory procedures in the week prior to actual data collection. The subjects performed four tests under S and P conditions on separate days over 2 weeks. These included measurement of maximal O2 uptake (VO2max) (cycle ergometry) with assessment of pulmonary function before and after, a submaximal (90% of ventilatory threshold) square-wave work transition from a base of unloaded cycling, a 60-s modified Wingate test, and a simulated 20 km time trial. No significant differences were observed in any of the dependent variables related to aerobic endurance or cycling performance between the S and P conditions. These results support other findings that an acute dose (400 micrograms) of S has no performance-enhancing properties.

Genetic dissection of carotenoid synthesis in arabidopsis defines plastoquinone as an essential component of phytoene desaturation.

Carotenoids are C40 tetraterpenoids synthesized by nuclear-encoded multienzyme complexes located in the plastids of higher plants. To understand further the components and mechanisms involved in carotenoid synthesis, we screened Arabidopsis for mutations that disrupt this pathway and cause accumulation of biosynthetic intermediates. Here, we report the identification and characterization of two nonallelic albino mutations, pds1 and pds2 (for phytoene desaturation), that are disrupted in phytoene desaturation and as a result accumulate phytoene, the first C40 compound of the pathway. Surprisingly, neither mutation maps to the locus encoding the phytoene desaturase enzyme, indicating that the products of at least three loci are required for phytoene desaturation in higher plants. Because phytoene desaturase catalyzes an oxidation reaction, it has been suggested that components of an electron transport chain may be involved in this reaction. Analysis of pds1 and pds2 shows that both mutants are plastoquinone and tocopherol deficient, in addition to their inability to desaturate phytoene. Separate steps of the plastoquinone/tocopherol biosynthetic pathway are affected by these two mutations. The pds1 mutation affects the enzyme 4-hydroxyphenylpyruvate dioxygenase because it can be rescued by growth on the product but not the substrate of this enzyme, homogentisic acid and 4-hydroxyphenylpyruvate, respectively. The pds2 mutation most likely affects the prenyl/phytyl transferase enzyme of this pathway. Because tocopherol-deficient mutants in the green alga Scenedesmus obliquus can synthesize carotenoids, our findings demonstrate conclusively that plastoquinone is an essential component in carotenoid synthesis. We propose a model for carotenoid synthesis in photosynthetic tissue whereby plastoquinone acts as an intermediate electron carrier between carotenoid desaturases and the photosynthetic electron transport chain.

Production of A and C variants of staphylococcal beta-lactamase by methicillin-resistant strains of Staphylococcus aureus.

Most methicillin-resistant Staphylococcus aureus (MRSA) strains produce beta-lactamase. To determine whether this enzyme(s) is identical to one or more of the four beta-lactamases produced by methicillin-susceptible strains, the beta-lactamases of 50 MRSA isolates were typed by using substrate profile analysis. Forty type A, no type B, ten type C, and no type D beta-lactamase-producing strains were identified. The beta-lactamase inhibitor sulbactam reduced the MICs of beta-lactamase-labile antibiotics, including ampicillin, penicillin G, and cefazolin, for type A and type C MRSA strains.

The intron of Arabidopsis thaliana polyubiquitin genes is conserved in location and is a quantitative determinant of chimeric gene expression.

We have isolated and determined DNA sequence for the 5'-flanking regions of three Arabidopsis thaliana polyubiquitin genes, UBQ3, UBQ10, and UBQ11. Comparison to cDNA sequences revealed the presence of an intron in the 5'-untranslated region at the same position immediately upstream of the initiator methionine codon in each of the three genes. An intron at this position is also present in two sunflower and two maize polyubiquitin genes. An intron is also found in the 5'-untranslated regions of several animal polyubiquitin genes, although the exact intron position is not conserved among them, and none are in the same position as those in the higher plant polyubiquitin genes. Chimeric genes containing the 5'-flanking regions of UBQ3, UBQ10, and UBQ11 in front of the coding regions for the reporter enzyme Escherichia coli beta-glucuronidase (GUS) were constructed. When introduced transiently into Arabidopsis leaves via microprojectile bombardment, all resulted in readily detectable levels of GUS activity that were quantitatively similar. The introns of UBQ3 and UBQ10 in the corresponding promoter fragments were removed by replacement with flanking cDNA sequences and chimeric genes constructed. These constructs resulted in 2.5- to 3-fold lower levels of marker enzyme activity after transient introduction into Arabidopsis leaves. The UBQ10 promoter without the 5' intron placed upstream of firefly luciferase (LUX) resulted in an average of 3-fold lower LUX activity than from an equivalent construct with the UBQ10 intron. A UBQ3 promoter cassette was constructed for the constitutive expression of open reading frames in dicot plants and it produced readily detectable levels of GUS activity in transient assays.

Placental permeability for morphine-3-beta-D-glucuronide in the guinea pig.

The fetal Vd and the PS of the major metabolite of morphine, M3G, were studied in pregnant guinea pigs during the last half of gestation. Fetal Vd was determined to be 0.334 ml/g of fetal plus placental weight and did not vary as the gestation progressed. The mean +/- s.e. PS for M3G was 3.7 +/- 0.3 X 10(-5) ml/sec/g and was independent of anaesthesia. This value was consistent with previous studies of other hydrophilic compounds, indicating that diffusion governs the placental passage of M3G. The PS value increased with increasing fetal weight which was consistent with structural changes in the guinea pig placenta as fetal age progresses in late gestation.