Cardiopulmonary and cerebrovascular acclimatization in children and adults at 3800 m.

Maturational differences exist in cardiopulmonary and cerebrovascular function at sea-level, but the impact of maturation on acclimatization responses to high altitude is unknown. Ten children (9.8 ± 2.5 years) and 10 adults (34.7 ± 7.1 years) were assessed at sea-level (BL), 3000 m and twice over 4 days at 3800 m (B1, B4). Measurements included minute ventilation ( V ̇ E ${\dot{V}}_{\rm{E}}$ ), end-tidal partial pressures of oxygen ( P ETO 2 ${P}_{{\rm{ETO}}_{\rm{2}}}$ ) and carbon dioxide, echocardiographic assessment of pulmonary artery systolic pressure (PASP) and stroke volume (SV) and ultrasound assessment of blood flow through the internal carotid and vertebral arteries was performed to calculate global cerebral blood flow (gCBF). At 3000 m, V ̇ E ${\dot{V}}_{\rm{E}}$ was increased from BL by 19.6 ± 19.1% (P = 0.031) in children, but not in adults (P = 0.835); SV was reduced in children (-11 ± 13%, P = 0.020) but not adults (P = 0.827), which was compensated for by a larger increase in heart rate in children (+26 beats min-1 vs. +13 beats min-1 , P = 0.019). Between B1 and B4, adults increased V ̇ E ${\dot{V}}_{\rm{E}}$ by 38.5 ± 34.7% (P = 0.006), while V ̇ E ${\dot{V}}_{\rm{E}}$ did not increase further in children. The rise in PASP was not different between groups; however, ∆PASP from BL was related to ∆ P ETO 2 ${P}_{{\rm{ETO}}_{\rm{2}}}$ in adults (R2  = 0.288, P = 0.022), but not children. At BL, gCBF was 43% higher in children than adults (P = 0.017), and this difference was maintained at high altitude, with a similar pattern and magnitude of change in gCBF between groups (P = 0.845). Despite V ̇ E ${\dot{V}}_{\rm{E}}$ increasing in children but not adults at a lower altitude, the pulmonary vascular and cerebrovascular responses to prolonged hypoxia are similar between children and adults. KEY POINTS: Children have different ventilatory and metabolic requirements from adults, which may present differently in the pulmonary and cerebral vasculature upon ascent to high altitude. Children (ages 7-14) and adults (ages 23-44) were brought from sea level to high altitude (3000 to 3800 m) and changes in ventilation, pulmonary artery systolic pressure (PASP) and cerebral blood flow (CBF) were assessed over 1 week. Significant increases in ventilation and decreases in left ventricle stroke volume were observed at a lower altitude in children than adults. PASP and CBF increased by a similar relative amount between children and adults at 3800 m. These results help us better understand age-related differences in compensatory responses to prolonged hypoxia in children, despite similar changes in pulmonary artery pressure and CBF between children and adults.

Trunk muscle training, posture fatigue, and performance in laparoscopic surgery.

PURPOSE: To investigate the effect of trunk muscle endurance training on the perception of back postural fatigue and performance of a laparoscopic task. MATERIALS AND METHODS: Thirty-one medical students (28 men and 13 women) with no laparoscopic surgical experience were randomly assigned to either a training group or a control group. Participants in the training group underwent a 6-week, 18-session trunk (abdominal and back muscle) endurance training program, whereas participants in the control group did not. Performance by all participants was assessed on a simulated laparoscopic task under varying conditions of low back postural fatigue, both before and after the training program. RESULTS: Participants in the training group showed significant improvements in trunk endurance after the 6-week, trunk endurance training program (P<0.05), whereas those in the control group did not. The improvements in trunk endurance were accompanied by significantly reduced perceptions of discomfort (P< .001) and fewer errors during performance of the laparoscopic task. (P<0.02), whereas no significant changes occurred for the control group (P>0.05). CONCLUSION: Increasing trunk endurance can increase postural fatigue and discomfort during simulated laparoscopic tasks, which may assist in the management of errors during laparoscopy.

MiAMP1, a novel protein from Macadamia integrifolia adopts a Greek key beta-barrel fold unique amongst plant antimicrobial proteins.

MiAMP1 is a recently discovered 76 amino acid residue, highly basic protein from the nut kernel of Macadamia integrifolia which possesses no sequence homology to any known protein and inhibits the growth of several microbial plant pathogens in vitro while having no effect on mammalian or plant cells. It is considered to be a potentially useful tool for the genetic engineering of disease resistance in transgenic crop plants and for the design of new fungicides. The three-dimensional structure of MiAMP1 was determined through homonuclear and heteronuclear ((15)N) 2D NMR spectroscopy and subsequent simulated annealing calculations with the ultimate aim of understanding the structure-activity relationships of the protein. MiAMP1 is made up of eight beta-strands which are arranged in two Greek key motifs. These Greek key motifs associate to form a Greek key beta-barrel. This structure is unique amongst plant antimicrobial proteins and forms a new class which we term the beta-barrelins. Interestingly, the structure of MiAMP1 bears remarkable similarity to a yeast killer toxin from Williopsis mrakii. This toxin acts by inhibiting beta-glucan synthesis and thereby cell wall construction in sensitive strains of yeast. The structural similarity of MiAMP1 and WmKT, which originate from plant and fungal phyla respectively, may reflect a similar mode of action.

Insect peptides with improved protease-resistance protect mice against bacterial infection.

At a time of the emergence of drug-resistant bacterial strains, the development of antimicrobial compounds with novel mechanisms of action is of considerable interest. Perhaps the most promising among these is a family of antibacterial peptides originally isolated from insects. These were shown to act in a stereospecific manner on an as-yet unidentified target bacterial protein. One of these peptides, drosocin, is inactive in vivo due to the rapid decomposition in mammalian sera. However, another family member, pyrrhocoricin, is significantly more stable, has increased in vitro efficacy against gram-negative bacterial strains, and if administered alone, as we show here, is devoid of in vitro or in vivo toxicity. At low doses, pyrrhocoricin protected mice against Escherichia coli infection, but at a higher dose augmented the infection of compromised animals. Analogs of pyrrhocoricin were, therefore, synthesized to further improve protease resistance and reduce toxicity. A linear derivative containing unnatural amino acids at both termini showed high potency and lack of toxicity in vivo and an expanded cyclic analog displayed broad activity spectrum in vitro. The bioactive conformation of native pyrrhocoricin was determined by nuclear magnetic resonance spectroscopy, and similar to drosocin, reverse turns were identified as pharmacologically important elements at the termini, bridged by an extended peptide domain. Knowledge of the primary and secondary structural requirements for in vivo activity of these peptides allows the design of novel antibacterial drug leads.

Prepubescents' ventilatory responses to exercise with reference to sex and body size.

STUDY OBJECTIVES: To examine the ventilatory responses of prepubescent children to submaximal and peak exercise using appropriate allometric modeling to control for differences in body size. DESIGN: Cross-sectional study of a representative sample of children. SETTING: Middle schools (8 to 11 years) in Exeter, UK. PARTICIPANTS: We studied 101 boys and 76 girls aged 11.1 (0.4) years and classified Tanner stage 1 for pubic hair (no true pubic hair). MEASUREMENTS: At rest: stature, mass, sum of skinfolds, hemoglobin concentration, FVC, and FEV1. During treadmill exercise at 7, 8, 9, and 10 km/h, and at peak exercise: oxygen uptake (VO2), minute ventilation (VE), tidal volume (VT), and respiratory frequency (Rf). RESULTS: At peak exercise, boys' VO2, VE, and VT were significantly (p<0.01) higher than girls' values and remained so even when the influence of body size was controlled using allometric principles. There were no significant (p>0.05) sex differences in Rf or the ratios VT/FVC or VE/VO2. When data were compared at the same relative exercise intensity (ie, 70 to 75% or 80 to 85% peak VO2), no significant (p>0.05) sex differences in Rf, VT/FVC, or VE/VO2 were detected. Boys' higher (p<0.001) VO2 values were reflected by their higher VE which remained higher than values for girls at both submaximal levels even when the influence of body size was covaried out. CONCLUSIONS: Prepubescent boys demonstrate higher peak VO2 than girls and this is supported by a higher VE and VT, even when the influence of body size is accounted for using allometry. Other ventilatory responses to both peak exercise and exercise at the same relative intensity are remarkably similar in both boys and girls.

Active video games and physical activity recommendations: a comparison of the Gamercize Stepper, XBOX Kinect and XaviX J-Mat.

OBJECTIVES: The current study was designed to evaluate the intensity levels of three exergames and determine the association with physical activity recommendations that correspond to specific outcomes. The variation in cardiovascular responses between the three exergames was also examined. DESIGN: We employed a cross-sectional laboratory design. METHODS: We recruited 18 girls to participate in a peak VO2 test and to play Gamercize, Kinect River Rush, XaviX J-Mat at three separate exergaming sessions. Linear regression equations of heart rate and percentage of peak VO2 were calculated for each participant to determine the intensity of exergame play. Differences in intensity between the three exergames and time spent in the recommended moderate (heart rate at ≥ 55% peak VO2) and vigorous (heart rate at ≥ 75% peak VO2) intensity levels were analyzed. We calculated the coefficient of variation for the mean heart rate to determine the difference in variance in heart rate values for the three exergames. RESULTS: When playing Gamercize and Kinect the girls did not play at recommended moderate or vigorous levels. Although the girls did not play at vigorous levels when playing XaviX J-Mat, our results showed that when playing XaviX J-Mat they did play at moderate intensity levels. No significant differences in the coefficient of variation between the three exergames were apparent. CONCLUSIONS: If active gaming is to be used to increase physical activity levels then individual differences in levels of exertion and specific activity recommendations need to be taken into consideration.

Improving aerobic power in primary school boys: a comparison of continuous and interval training.

The purpose of this study was to assess whether the magnitude of change in aerobic power was different in boys (mean age 10.25 +/- 0.50 y) who followed a high-intensity interval training protocol, compared to those who followed a moderate-intensity continuous training protocol. Boys were assigned to either a control group (n = 15), a continuous training group (n = 10), or an interval training group (n = 10). They completed peak oxygen uptake tests at baseline and following an 8-week training period. The control group continued with normal activity habits, whilst the continuous training group followed a 20-minute steady-state cycle protocol at 80-85 % of the maximal heart rate, and the interval training group completed 30-s sprints on a cycle ergometer, interspersed with active rest periods. The two training protocols were designed to incur similar cardiovascular work over the 20 minutes of each training session. Significant increases (p < 0.05) in peak oxygen uptake were noted for both the interval and continuous training groups. The interval training group showed marked pre- to post-increases in both peak oxygen pulse, oxygen pulse at the ventilatory threshold, and ventilatory threshold that were not apparent in the continuous group boys. It would appear that a high-intensity interval protocol confers a different training effect in comparison to continuous steady-state training in boys. Possible mechanisms that underpin these adaptations may include increased blood volume and a concomitant adjustment in stroke volume.

Effect of training on the aerobic power and anaerobic performance of prepubertal girls.

The purpose of this study was to investigate the effects of two, three times a week, 8-week training programmes on the aerobic power and anaerobic performance of 30 prepubescent girls, with a mean age of 9.6 y. Peak oxygen uptake assessed by an incremental discontinuous treadmill test, and peak power in 5 s and mean power over 30 s estimated from a Wingate anaerobic test were used as the criterion measures. Twelve girls trained using a continuous cycle ergometer programme, 11 girls followed a sprint running programme and the control group consisted of 7 girls. Both training groups significantly (p < 0.05) increased their peak oxygen uptake and peak power in 5 s. However, the increase reported here are lower than those generally observed in adolescents following training. The control group demonstrated no significant (p > 0.05) change in either variable. No significant (p > 0.05) changes in mean power over 30 s were observed in any group.

Sigmoidity of the ventricular septum revisited: progression in early adulthood, predominance in men, and independence from cardiac mass.

Meager quantitative anatomic information is available regarding the orientation of the left ventricular outflow tract (LVOT) relative to the ventricular septum (VS), or the relation of these variables to patient age, sex, or other cardiac features. We studied 57 formaldehyde-fixed adult human heart specimens at autopsy from 29 men and 28 women ranging in age from 20 to 91 years (mean age = 66 years). Blinded measurement of 10 morphologic parameters, repeated on 2 occasions, included anatomic indexes of VS sigmoidity (angle between the aortic and the mitral plane). While sigmoidity was not correlated with patient height, body weight, body mass index, cardiac mass (or presence of systemic hypertension), cause of death, or VS length, it was significantly (p less than 0.05) progressive through age groups 20-39 years, 40-59, 60-79 and greater than or equal to 80. The overall correlation of aortic-mitral plane angle (A-MPA) with age was +0.59 (P less than 0.0001), a relationship also noted within each age group. The mean A-MPA increased from 118 degrees to 127, 131 and 134 considering all specimens. Men had consistently greater sigmoidity than women. The steady increase in sigmoidity of the ventricular septum from early adulthood may alter clinical conclusions about pathologic conditions which are based on septal curvature or prominence.

Aerobic fitness of prepubescent children.

This study was designed to enhance understanding of the assessment and interpretation of the aerobic fitness of prepubertal children. Written informed consent to participate was obtained from 70% of the children in year six of the 15 state schools in the city of Exeter. Twenty-five per cent of the eligible children in each school were randomly selected from those who volunteered. The data reported here are those obtained from the 111 boys (11.1 SD 0.4 years) and 53 girls (10.9 SD 0.3 years) classified as Tanner stage 1 in both pubic hair rating and either genitalia rating (boys) or breast rating (girls). Peak oxygen uptake (peak VO2) was determined using a discontinuous, incremental protocol on a treadmill. Only a minority of children demonstrated a levelling-off or plateau in VO2 despite an increase in exercise intensity. There was no evidence to suggest that the children who demonstrated a VO2 plateau had significantly (p < 0.05) higher peak VO2, peak heart rate, peak respiratory exchange ratio or peak blood lactate than those children who did not demonstrate a plateau in VO2. These findings indicate that a VO2 plateau should not be used as a requirement for defining a maximal exercise test with prepubertal children. Boys had a significantly (p < 0.01) higher peak VO2 than girls, whether expressed in 1.min-1 (1.78 vs 1.46) or in relation to body mass (51 vs 45 ml.kg-1.min-1). The results compare favourably with those of similarly aged children from other countries, but why prepubescent boys have significantly higher (13.3%) mass-related peak VO2 than prepubescent girls is not readily apparent. Although conventional, the expression of peak VO2 as per body mass ratio may not adequately partition out body-size differences. The influence of body mass was therefore removed using a linear adjustment scaling model and a log-linear model, but the boys' peak VO2 remained significantly (p < 0.01) higher than the girls' peak VO2 with the difference now being 16.0% and 16.2%, respectively.

Conformational studies by NMR of the antimicrobial peptide, drosocin, and its non-glycosylated derivative: effects of glycosylation on solution conformation.

Drosocin is a cationic 19 amino acid peptide secreted by Drosophila in response to septic injury. The sequence (GKPRPYSPRPTSHPRPIRV) contains six Pro and four Arg residues which are incorporated into three repeated triplet sequences Pro-Arg-Pro. The peptide is glycosylated at Thr11 and has potent antimicrobial activity. This activity is markedly reduced on deglycosylation, but a structural basis for this has not been previously established. In the current study, the solution conformations of drosocin and its non-glycosylated derivative were determined by NMR spectroscopy and structure calculations. The NMR and structure studies showed that the peptides have significant populations of essentially random coil conformations in aqueous solution. Addition of 50% trifluoroethanol causes the development of small populations of folded conformations, mainly in the form of turns. In particular, turn elements occur near residues 4-7, 10-13, 17, and 18. No substantial difference was detected in the predominantly random coil conformation of the glycosylated and non-glycosylated forms, but there are subtle differences in the small populations of folded conformers. In particular, the turn at residues 10-13 tends toward a more extended structure on glycosylation, while there is some tightening of the downstream turn at residues 17 and 18. There are a significant number of nuclear Overhauser enhancement contacts between the sugar moiety and the peptide near the glycosylation site, consistent with a close association between them. Despite this close association, the pKa of H13, which is proximate to the glycosylation site, was found to be unaffected by glycosylation.

Three-dimensional structure of RK-1: a novel alpha-defensin peptide.

NMR spectroscopy and simulated annealing calculations have been used to determine the three-dimensional structure of RK-1, an antimicrobial peptide from rabbit kidney recently discovered from homology screening based on the distinctive physicochemical properties of the corticostatins/defensins. RK-1 consists of 32 residues, including six cysteines arranged into three disulfide bonds. It exhibits antimicrobial activity against Escherichia coli and activates Ca(2+) channels in vitro. Through its physicochemical similarity, identical cysteine spacing, and linkage to the corticostatins/defensins, it was presumed to be a member of this family. However, RK-1 lacks both a large number of arginines in the primary sequence and a high overall positive charge, which are characteristic of this family of peptides. The three-dimensional solution structure, determined by NMR, consists of a triple-stranded antiparallel beta-sheet and a series of turns and is similar to the known structures of other alpha-defensins. This has enabled the definitive classification of RK-1 as a member of this family of antimicrobial peptides. Ultracentrifuge measurements confirmed that like rabbit neutrophil defensins, RK-1 is monomeric in solution, in contrast to human neutrophil defensins, which are dimeric.

Chemical synthesis, characterization and activity of RK-1, a novel alpha-defensin-related peptide.

The 32-residue peptide, RK-1, a novel kidney-derived three disulfide-bonded member of the antimicrobial alpha-defensin family, was synthesized by the continuous flow Fmoc-solid phase method. The crude, cleaved and S-reduced linear peptide was both efficiently folded and oxidized in an acidic solution of aqueous dimethyl sulfoxide. Following purification of the resulting product, it was shown by a variety of analytical techniques, including matrix assisted laser desorption time of flight mass spectrometry, to possess a very high degree of purity. The disulfide bond pairing of the synthetic peptide was determined by 1H-NMR spectroscopy and confirmed to be a Cys1-Cys6, Cys2-Cys4, Cys3-Cys5 arrangement similar to other mammalian alpha-defensin peptides. The synthetic RK-1 was also shown to inhibit the growth of Escherichia coli type strain NCTC 10418.

Purification and characterization of a plant antimicrobial peptide expressed in Escherichia coli.

MiAMP1 is a low-molecular-weight, cysteine-rich, antimicrobial peptide isolated from the nut kernel of Macadamia integrifolia. A DNA sequence encoding MiAMP1 with an additional ATG start codon was cloned into a modified pET vector under the control of the T7 RNA polymerase promoter. The pET vector was cotransformed together with the vector pSB161, which expresses a rare arginine tRNA. The peptide was readily isolated in high yield from the insoluble fraction of the Escherichia coli extract. The purified peptide was shown to have an identical molecular weight to the native peptide by mass spectroscopy indicating that the N-terminal methionine had been cleaved. Analysis by NMR spectroscopy indicated that the refolded recombinant peptide had a similar overall three-dimensional structure to that of the native peptide. The peptide inhibited the growth of phytopathogenic fungi in vitro in a similar manner to the native peptide. To our knowledge, MiAMP1 is the first antimicrobial peptide from plants to be functionally expressed in E. coli. This will permit a detailed structure-function analysis of the peptide and studies of its mode of action on phytopathogens.