Dietary polyunsaturated fatty acids in asthma- and exercise-induced bronchoconstriction.

Despite progress that has been made in the treatment of asthma, the prevalence and burden of this disease has continued to increase. While pharmacological treatment of asthma is usually highly effective, medications may have significant side effects or exhibit tachyphylaxis. Alternative therapies for treatment that reduce the dose requirements of pharmacological interventions would be beneficial, and could potentially reduce the public health burden of this disease. Ecological and temporal data suggest that dietary factors may have a role in recent increases in the prevalence of asthma. A possible contributing factor to the increased incidence of asthma in Western societies may be the consumption of a proinflammatory diet. In the typical Western diet, 20- to 25-fold more omega (n)-6 polyunsaturated fatty acids (PUFA) than n-3 PUFA are consumed, which promotes the release of proinflammatory arachidonic acid metabolites (leukotrienes and prostanoids). This review will analyze the evidence for the health effects of n-3 PUFA in asthma- and exercise-induced bronchoconstriction (EIB). While clinical data evaluating the effect of omega-3 fatty acid supplementation in asthma has been equivocal, it has recently been shown that fish oil supplementation, rich in n-3 PUFA, reduces airway narrowing, medication use, and proinflammatory mediator generation in nonatopic elite athletes with EIB. These findings are provocative and suggest that dietary fish oil supplementation may be a viable treatment modality and/or adjunct therapy in asthma and EIB.

Effects of montelukast on airway narrowing from eucapnic voluntary hyperventilation and cold air exercise.

BACKGROUND: Exercise induced bronchoconstriction (EIB) is common in elite athletes. Eucapnic voluntary hyperventilation (EVH) is a laboratory test recommended for the identification of EIB in athletes, secondary to a field exercise challenge. Montelukast attenuates EIB, but its protective effect against airway narrowing from EVH has not been investigated. OBJECTIVE: To examine the effectiveness of montelukast after exercise and after EVH. METHODS: A randomised, placebo controlled, double blind, crossover study was performed with 11 physically active EIB positive subjects (eight men, three women; mean (SD) age 22.8 (6.8) years). Six hours before each of the following challenges 10 mg montelukast or placebo was ingested: (a) a six minute, cold air (-3 degrees C) maximal effort work accumulation cycle ergometer exercise; (b) EVH, breathing 5% CO(2) compressed air at 85% maximal voluntary ventilation for six minutes. Spirometry was performed before and 5, 10, and 15 minutes after the challenge. At least 48 hours was observed between challenges. RESULTS: No differences in forced expiratory volume in one second (FEV(1)) were found after the two challenges. Exercise and EVH resulted in falls in FEV(1) of 22.4 (18.0) and 25.6 (16.8) respectively. Falls in FEV(1) after montelukast were less than after placebo (10.6 (10.6) and 14.3 (11.3) after exercise and EVH respectively; p<0.05). Montelukast provided protection against bronchoconstriction (59% and 53%; p<0.05) for eight exercising subjects and 10 EVH subjects; no protection was afforded for three exercising and one EVH challenged subject. CONCLUSIONS: Both exercise and EVH were potent stimuli of airway narrowing. A single dose of montelukast provided reasonable protection in attenuating bronchoconstriction from either exercise or EVH. The similar protection by montelukast suggests that EVH is a suitable laboratory surrogate for EIB evaluation.

Overexpression of simian virus 40 small-T antigen blocks centrosome function and mitotic progression in human fibroblasts.

Recombinant adenoviruses that express high levels of the simian virus 40 (SV40) small-t (ST) antigen have been used to study the requirement for ST to drive cell cycle proliferation of confluent human diploid fibroblasts. This occurs when either large-T (LT) antigen or serum is added to provide a second signal. While cells readily completed S phase in these experiments, they were found to accumulate with 4N DNA content. Cellular and nuclear morphology, as well as the biochemical status of cyclin B complexes, showed that these cells entered mitosis but were blocked prior to mitotic metaphase. The defect appears to reflect an inability of cells overexpressing ST to form organized centrosomes that duplicate and separate normally during the cell cycle and, therefore, the absence of a mitotic spindle. The ability of ST to bind protein phosphatase 2A was required for this pattern, suggesting that altered phosphorylation of key centrosomal components may occur when ST is overexpressed. Although the possible significance of ST effects on the centrosome cycle is not fully understood, these findings suggest that ST could influence chromosomal instability patterns that are a hallmark of SV40-transformed cells and LT expression.

Muscle oxygen desaturation is related to whole body VO2 during cross-country ski skating.

Previous research has demonstrated that blood flow and subsequent O2 desaturation (OD) in exercising muscle is related to the static component during exercise. In speed skating, increased OD is dissociated from whole body VO2 and heart rate (HR) when the skater increases the static component by 'sitting low'. This phenomenon was evaluated in cross-country skiers by manipulating speed and incline during treadmill roller skiing. Eight male cross-country skiers (22.4 +/- 3.2 yrs old) randomly performed constant incline- and constant speed-based protocols in which increased load was manipulated in five 4min stages by treadmill incline or speed change, respectively. A strong relationship (r = 0.83) was observed between VO2 and % OD while blood volume change (deltaBV) was minimal. Unexpectedly, no HR/ VO2 or HR/OD shifts were observed between protocols. The % OD response, in relation to blood lactate values, during submaximal exercise was very similar to that of VO2. The lack of an observed greater desaturation at higher inclines suggests that the expected static load may be attenuated by an increased contribution of poling. The strong relationship of % OD to whole body VO2 may be attributed to O2 dissociation in the capillary bed of the muscle to meet aerobic energy demand and is independent of blood flow dynamics during cross-country ski skating.

The role of the SV40 ST antigen in cell growth promotion and transformation.

The simian virus 40 small-t (ST) antigen plays a key role in permissive and nonpermissive infections, increasing virus yields in lytic cycles of primate cells and enhancing the ability of large-T (LT) to transform rodent or even human cells. In the absence of ST, tumors in rodent model systems appear primarily in lymphoid and other proliferative tissues and transformation is reduced in several in vitro systems. The functions of ST largely reflect its binding and inhibition of protein phosphatase 2A, although a recently described dnaJ domain also contributes to its biology. The dnaJ domain is present in LT and a third early gene product, the 17kT protein, for which a potential role in transformation deserves further evaluation.

Self-reported symptoms and exercise-induced asthma in the elite athlete.

PURPOSE: The purpose of this study was to compare self-reported symptoms for exercise-induced asthma (EIA) to postexercise challenge pulmonary function test results in elite athletes. METHODS: Elite athletes (N = 158; 83 men and 75 women; age: 22 +/- 4.4 yr) performed pre- and post-exercise spirometry and were grouped according to postexercise pulmonary function decrements (PFT-positive, PFT-borderline, and PFT-normal for EIA). Before the sport/environment specific exercise challenge, subjects completed an EIA symptoms-specific questionnaire. RESULTS: Resting FEV1 values were above predicted values (114--121%) and not different between groups. Twenty-six percent of the study population demonstrated >10% postexercise drop in FEV1 and 29% reported two or more symptoms. However, the proportion of PFT-positive and PFT-normal athletes reporting two or more symptoms was not different (39% vs. 41%). Postrace cough was the most reported symptom, reported significantly more frequently for PFT-positive athletes (P < 0.05). Sensitivity/specificity analysis demonstrated a lack of effectiveness of self-reported symptoms to identify PFT-positive or exclude PFT-normal athletes. Postexercise lower limit reference ranges (MN-2SDs) were determined from normal athletes for FEV1, FEF25--75% and PEF to be -7%, -12.5%, and -18%, respectively. CONCLUSION: Although questionnaires provide reasonable estimates of EIA prevalence among elite cold-weather athletes, the use of self-reported symptoms for EIA diagnosis in this population will likely yield high frequencies of both false positive and false negative results. Diagnosis should include spirometry using an exercise/environment specific challenge in combination with the athlete's history of asthma symptoms.

Hemoglobin/myoglobin oxygen desaturation during Alpine skiing.

PURPOSE: To investigate muscle blood volume (BV) change and hemoglobin/myoglobin oxygen desaturation (OD) during simulated giant slalom (GS) and slalom (SL) Alpine ski racing. METHODS: Joint angle, BV, OD, and heart rate (HR) were evaluated during GS and SL events in 30 junior elite skiers ages 9--17 yr (13.5 +/- 2.3). Subjects were stratified by ski class and age: group I, J1 and J2, ages 15--18 yr (16.8 +/- 0.8); group II, J3, 13--14 yr (13.6 +/- 0.7); and group III, J4 and J5, 9--12 yr (11.5 +/- 1.2). Near-infrared spectrophotometry (NIRS) was used to measure BV and OD in the capillary bed of the vastus lateralis during trials. Maximal OD was determined during thigh cuff ischemia (CI). Quadriceps cross-sectional area (CSA) was estimated by skin-fold and thigh circumference. RESULTS: Joint angles were smaller (P < 0.05) during GS than SL for ankle (83.8 +/- 11.9 degrees; 98.6 +/- 15.7 degrees ), knee (107.4 +/- 14.9 degrees; 118.3 +/- 18.0 degrees ), and hip (98.8 +/- 14.3 degrees; 107.5 +/- 16.2 degrees ). BV reduction from rest to peak exercise (Delta BV) was 30% greater (P < 0.05) during the GS than SL, whereas Delta OD was 33% greater (P < 0.05) during GS. Delta OD, relative to CI OD, was greater for all subjects during GS (79.2 +/- 3.7%) than SL (65.7 +/- 4.4%). This pattern continued within groups; group II displayed the greatest relative desaturation (82.9 +/- 7.6%). CSA was larger in older skiers (92.5 +/- 21.6; 72.5 +/- 12.3; 65.3 +/- 21.2 cm(2)) and correlated with Delta OD (P < 0.05). CONCLUSION: The larger reduction in BV (Delta BV change) and greater OD when skiers assumed lower posture during GS than SL may be related to greater effective static load secondary to higher percent of maximal voluntary contraction and is consistent with compromised blood flow to working muscle.

Critical role for SV40 small-t antigen in human cell transformation.

Defining the ability of simian virus 40 (SV40) to transform human cells has become of even greater importance with the increased understanding that this virus may play a role in some human malignancies. This report documents the requirement for viral small-t (ST) antigen in large-T (LT)-driven transformation of primary fibroblasts, a requirement that cannot be met by a well-known oncogene, c-Ha-ras (EJ-ras), which can cooperate with LT in rodent systems. The cellular gene telomerase is not essential for transformation, although transformed clones are not immortal without it. Similarly, an immortal mesothelial cell line has been developed using LT and telomerase. Immortalized mesothelial cells are morphologically normal, but can be transformed by introduction of ST, or ST + ras, but not by ras alone. It is likely that ST will be required along with LT for transformation of most human cell types.

Incidence of exercise-induced bronchospasm in Olympic winter sport athletes.

PURPOSE: The purpose of this project was to determine the incidence of exercise-induced bronchospasm (EIB) among U.S. Olympic winter sport athletes. METHODS: Subjects included female and male members of the 1998 U.S. Winter Olympic Team from the following sports: biathlon, cross-country ski, figure skating, ice hockey, Nordic combined, long-track speedskating, and short-track speedskating. Assessment of EIB was conducted in conjunction with an "actual competition" (Olympic Trials, World Team Trials, World Cup Event, U.S. National Championships) or a "simulated competition" (time trial, game), which served as the exercise challenge. Standard spirometry tests were performed preexercise and at 5, 10, and 15 min postexercise. An athlete was considered EIB-positive based on a postexercise decrement in FEV1 > or = 10%. RESULTS: For the seven sports evaluated on the 1998 U.S. Winter Olympic Team, the overall incidence of EIB across all sports and genders was 23%. The highest incidence of EIB was found in cross-country skiers, where 50% of the athletes (female = 57%; male = 43%) were diagnosed with EIB. Across the seven sports evaluated, the prevalence of EIB among the female and male athletes was 26% and 18%, respectively. Among those individuals found to be EIB-positive were athletes who won a team gold medal, one individual silver medal, and one individual bronze medal at the Nagano Winter Olympics. CONCLUSIONS: These data suggest that: 1) EIB is prevalent in several Olympic winter sports and affects nearly one of every four elite winter sport athletes; 2) the winter sport with the highest incidence of EIB is cross-country skiing; 3) in general, EIB is more prevalent in female versus male elite winter sport athletes; and 4) athletes may compete successfully at the international level despite having EIB.

Exercise-induced asthma screening of elite athletes: field versus laboratory exercise challenge.

PURPOSE: The purpose of this study was to compare a laboratory based exercise challenge (LBC) to a field based exercise challenge (FBC) for pulmonary function test (PFT) exercise-induced asthma (EIA) screening of elite athletes. METHODS: Twenty-three elite cold weather athletes (14 men, 9 women) PFT positive for EIA (FBC screened) served as subjects. Twenty-three gender and sport matched controls (nonasthmatics) were randomly selected to establish PFT reference values for normal elite athletes. Before FBC, athletes completed a medical history questionnaire for EIA symptoms. FBC evaluations consisted of baseline spirometry, actual or simulated competition, and 5, 10, and 15 min postexercise spirometry. PFT positive athletes were evaluated in the laboratory using an exercise challenge simulating race intensity (ambient conditions: 21 degrees C, 60% relative humidity). PFT procedures were identical to FBC. RESULTS: 91% of PFT positive and 48% of PFT normal athletes reported at least one symptom of EIA, with postrace cough most frequent. Baseline spirometry was the same for PFT positives and normal controls. Lower limit reference range (MN - 2 SD) of FEV1 for controls suggests that postexercise decrements of greater than approximately -7% indicate abnormal airway response in this population. Exercise time duration did not effect bronchial reactivity; 78% of FBC PFT positives were PFT normal post-LBC. CONCLUSION: Self-reported symptoms by elite athletes are not reliable in identifying EIA. Reference range criterion for FEV1 decrement in the elite athlete postexercise contrasts current recommended guidelines. Moreover, a large number of false negatives may occur in this population if EIA screening is performed with inadequate exercise and environmental stress.

The leader polypeptide of Theiler's murine encephalomyelitis virus is required for the assembly of virions in mouse L cells.

Deletion of the entire leader polypeptide of the GDVII strain of Theiler's murine encephalomyelitis virus (TMEV) results in the production of an attenuated virus that grows in baby hamster kidney (BHK) cells but cannot grow at all in mouse L-929 cells. This study examined the reasons for the failure of dl-L, the GDVII variant that lacks the leader polypeptide, to grow in mouse cells. At low multiplicities of infection, it was difficult to detect any viral proteins in mouse cells. However, levels of positive- and negative-strand RNA molecules were only moderately reduced in these infections. Viral RNA showed no major defect in translatability, as the mutant viral RNA was nearly as efficient as that of the wild-type (WT) virus in directing protein synthesis in vitro in assays using extracts prepared from mouse L cells. Viral protein synthesis was detected in dl-L-infected mouse cells as multiplicities of infection were increased and approached the levels observed in WT infections. Despite this, there was a total lack of virus production in high-multiplicity infections, and this was found to correlate with the failure of viral proteins and early virion precursors to assemble into virions in mouse cells. Thus, the inability of dl-L to grow in mouse cells reflects complex effects on various stages of the virus infection but is primarily a defect in virus assembly.

Evidence for restricted muscle blood flow during speed skating.

INTRODUCTION: We have previously hypothesized restricted muscle blood flow during speed skating, secondary to the high intramuscular forces intrinsic to the unique posture assumed by speed skaters and to the prolonged duty cycle of the skating stroke. METHODS: To test this hypothesis, we studied speed skaters (N = 10) during submaximal and maximal cycling and in-line skating, in both low (knee angle = 107 degrees) and high (knee angle = 112 degrees) skating positions (CE vs SkL vs SkH). Supportive experiments evaluated muscle desaturation and lactate accumulation during on-ice speed skating and muscle desaturation during static exercise at different joint positions. RESULTS: Consistent with the hypothesis were reductions during skating in VO2peak (4.28 vs 3.83 vs 4.26 L x min(-1)), the VO2 at 4 mmol x L(-1) blood lactate (3.38 vs 1.93 vs 3.31 L x min(-1)), and cardiac output during maximal exercise (33.2 vs 25.3 vs 25.6 L x min(-1)). The reduction in maximal cardiac output was not attributable to differences in HRmax (197 vs 192 vs 193 b x min(-1)) but to a reduction in SVmax (172 vs 135 vs 134 mL x beat(-1)). The reduction in SV appeared to be related to an increased calculated systemic vascular resistance (354 vs 483 vs 453 dynes x s(-1) x cm(-1)). During maximal skating there was also a greater % O2 desaturation of the vastus lateralis based on near infrared spectrophotometry (50.3 vs 74.9 vs 60.4% of maximal desaturation during cuff ischemia). The results were supported by greater desaturation with smaller knee angles during static exercise and by greater desaturation and accelerated blood lactate accumulation during on-ice speed skating in the low vs high position. The results of this study support the hypothesis that physiological responses during speed skating are dominated by restriction of blood flow, attributable either to high intramuscular forces, the long duty cycle of the skating stroke, or both.

Upper body power comparison between groups of cross-country skiers and runners.

The purpose of this study was to evaluate relationships between upper body power (UBP) and cross-country ski skating race velocity (R) in a cross sectional representation of cross-country skiers. Additionally, cross-country skiers' UBP was compared to UBP of distance runners. Participants (n = 195) were tested on a Street Arm Ergometer for UBP using a ramped maximal UBP test simulating a double poling motion. A strong relationship (r = 0.89) between UBP and RV in skiers was determined. High school skiers were separated into slow and fast groups based on reported RV; significant differences (p < 0.05) in both UBP was found to be an effective partial predictor of RV, independent of gender. Mean UBP for the cross-country runners was 46% of mean UBP for the cross-country skiers. With UBP contributing so much to performance in cross-country ski racing, it is recommended that cross-country skiers focus a large portion of their training on the specific development of that fitness component.

The simian virus 40 small-t and large-T antigens jointly regulate cell cycle reentry in human fibroblasts.

Focus formation in human diploid fibroblasts (HDF cells) is known to require both the simian virus 40 (SV40) large-T and small-t antigens. Similarly, both SV40 proteins were required to stimulate confluent, density-arrested HDF cells to reenter the cell cycle. This study used defective recombinant adenoviruses to examine the roles of the individual SV40 proteins in altering specific steps in the cell cycle. Small-t antigen and, to a lesser extent, large-T antigen increased the level of the S phase cyclin cyclin A but without increasing the activity of associated cyclin kinases unless the two SV40 proteins were coexpressed. The absence of kinase activity reflected the presence in density-arrested cells of high levels of the cyclin-dependent kinase inhibitors p21(WAF1) and p27(KIP1). We report here that expression of SV40 large-T antigen reduced levels of p21(WAF1), while expression of small-t antigen was required to decrease p27(KIP1). The separate effects of large-T and small-t antigens on these two inhibitors may explain the joint requirement for the two proteins to drive cell cycle reentry of HDF cells and ultimately transform these cells.

Cell cycle progression in monkey cells expressing simian virus 40 small t antigen from adenovirus vectors.

The simian virus 40 small t antigen (small-t) is required for optimal viral replication and transformation, especially during the infection of nondividing cells, suggesting that the function of small-t is to promote cell cycle progression. The mechanism through which small-t promotes cell growth reflects, in part, its binding and inhibition of protein phosphatase 2A (PP2A). The use of recombinant adenoviruses allows small-t expression in a majority of cells in a population, thus providing a convenient source of cells for biochemical analyses. In monkey kidney CV1 cells, small-t expressed from these adenovirus vectors activated the mitogen-activated protein kinase (MAPK) pathway, induced JNK activity, and increased AP-1 DNA-binding activity, all in a PP2A-dependent manner. Expression of small-t also caused an increase in the phosphorylation of the Na+/H+ antiporter, a mitogen-activated ion exchanger whose activity correlates with its phosphorylation. At least part of the antiporter phosphorylation induced by small-t reflected activation of the MAPK pathway, as suggested by results of assays using a chemical inhibitor of the MAPK-activating kinase, MEK. Finally, small-t expression from adenovirus vectors promoted efficient cell cycle progression by growth-arrested cells. These vectors should facilitate further analysis of effects of small-t on cell cycle mediators.

Small-t and large-T antigens cooperate to drive cell proliferation.

Optimal transformation efficiencies or tumour formation in certain target tissues require the SV40 small-t antigen in addition to the transforming protein, large-T. We have used two model systems in which small-t is required for transformation to roles of individual viral proteins in this process. These systems include anchorage-independent growth of rat fibroblasts and focus formation by primary human diploid fibroblasts. In both cases, large-T and small-t antigens work together to drive cell cycle induction. Thus, the need for both tumour antigens is apparent in the initial step of the transformation process, the stimulation of quiescent cells to enter the cell cycle.

Energy cost of rifle carriage in biathlon skiing.

PURPOSE: Since biathlon racing involves cross-country skiing while carrying a minimum weight 3.5-kg rifle, energy cost for rifle carriage could be significant to race outcome. The purposes of this study were to: 1) compare physiological measurements of biathletes roller skiing with and without rifle carriage, 2) compare energy cost of rifle carriage between women and men, 3) examine the relationship of cycle length and cycle rate to energy cost of rifle carriage, and 4) compare physiological measurements to calculated estimates of power requirements of rifle carriage. METHODS: We examined metabolic cost of rifle carriage during inclined roller ski-skating. National Team Biathletes (7 W, 8 M) performed treadmill roller ski-skating (2.46, 2.68, and 2.91 m.s-1, 8% incline, 5-min stages) with and without a 3.65 kg rifle. RESULTS: For W, HR, VO2, and VE during carriage were higher at all speeds, LA was higher at the fastest speed (P < 0.05). For M, VO2 and VE were higher at all speeds, HR and LA were higher at 2.68 and 2.91 m.s-1 (P < 0.05). Rifle mass as percent of body mass was different between W and M (6.6 +/- 0.7% vs 5.0 +/- 0.3%, P < 0.05). Percent increase in VO2 (2.1% and 1.3% per kg load; for W and M, respectively) was not different than rifle mass as percent body wt. Cycle length was related to increased VO2 and LA during rifle carriage for W (r = -0.59 and -0.70 to -0.85), and to LA for M (r = -0.66 to -0.83). CONCLUSION: The large range in cost between individuals (0.2 +/- 0.08 and 0.19 +/- 0.17 L.min-1; for W & M, respectively) suggests that individual economies for load carriage can be improved.

The amino-terminal transforming region of simian virus 40 large T and small t antigens functions as a J domain.

Simian virus 40 (SV40) encodes two proteins, large T antigen and small t antigen that contribute to virus-induced tumorigenesis. Both proteins act by targeting key cellular regulatory proteins and altering their function. Known targets of the 708-amino-acid large T antigen include the three members of the retinoblastoma protein family (pRb, p107, and p130), members of the CBP family of transcriptional adapter proteins (cap-binding protein [CBP], p300, and p400), and the tumor suppressor p53. Small t antigen alters the activity of phosphatase pp2A and transactivates the cyclin A promoter. The first 82 amino acids of large T antigen and small t antigen are identical, and genetic experiments suggest that an additional target(s) important for transformation interacts with these sequences. This region contains a motif similar to the J domain, a conserved sequence found in the DnaJ family of molecular chaperones. We show here that mutations within the J domain abrogate the ability of large T antigen to transform mammalian cells. To examine whether a purified 136-amino-acid fragment from the T antigen amino terminus acts as a DnaJ-like chaperone, we investigated whether this fragment stimulates the ATPase activity of two hsc70s and discovered that ATP hydrolysis is stimulated four- to ninefold. In addition, ATPase-defective mutants of full-length T antigen, as well as wild-type small t antigen, stimulated the ATPase activity of hsc70. T antigen derivatives were also able to release an unfolded polypeptide substrate from an hsc70, an activity common to DnaJ chaperones. Because the J domain of T antigen plays essential roles in viral DNA replication, transcriptional control, virion assembly, and tumorigenesis, we conclude that this region may chaperone the rearrangement of multiprotein complexes.