The redox reaction between cytochrome c (Cyt c) (P-551) and the blue copper protein azurin, both from Pseudomonas aeruginosa, was studied using the temperature-jump technique. Two relaxation times were observed in a mechanism assumed to involve three equilibria. The fast relaxation time (0.4 less than tau less than 8 ms) was ascribed to the electron exchange step. The slow relaxation time (tau congruent to 37 ms) was assigned to a conformational equilibrium of the reduced azurin that was coupled through the electron exchange step to a faster conformational equilibrium of the oxidized Cyt c (P551). But because the Cyt c (P551) isomerization, being very rapid, was uncoupled from the two slower equilibria, and was assumed to involve no spectral change, the amplitude of its relaxation time (tau congruent to 0.1 ms) would be zero. At 25 degrees C and pH 7.0 the rate constants for the oxidation and reduction of Cyt c (P551) by azurin were 6.1 X 10(6) and 7.8 X 10(6) M-1 s-1, respectively; for the formation and disappearance of the reactive conformational isomer of azurin they were 12 and 17 s-1, respectively. The rates for the Cyt c (P551) isomerization could only be estimated at approximately 10(4) s-1. The thermodynamic parameters of each reaction step were evaluated from the amplitudes of the relaxations and from Eyring plots of the rate constants. Measurements of the overall equilibrium constant showed it to be temperature independent (5-35 degrees C), i.e. deltaHtot = 0. This zero enthalpy change was found to be compatible with the enthalpies calculated for the individual steps. In the electron exchange equilibrium, the values of the activation enthalpies were two to three times higher than the values published for various low molecular weight reagents in their electron exchange with copper proteins, yet the rate of exchange between Cyt c (P551) and azurin was some hundreds of times faster. This was explained in terms of the measured positive or zero entropies of activation that could result from a high level of specificity between the proteins particularly in areas of complementary charges. The mechanism of electron transfer was considered as essentially an outer sphere reaction, of which the rate could be approximated by the Marcus theory.

One hundred fifty war veterans with ankylosing spondylitis were entered into a prospective study in 1947. In 1957, 142 were traced, and they have been reviewed periodically. Eighty‐one of these patients were still alive in 1980. Information was obtained from 67 (83%) of the survivors and 51 were reexamined. This report is based on the clinical findings in these 51 patients, who have a mean disease duration of 38 years. Forty‐seven (92%) were functioning well. The disease in 21 (41%) had progressed to cause severe spinal restriction. Of those, 12 had peripheral joint involvement early in their course and 9 had iritis. Seventy‐four percent of the patients who had mild spinal restriction after 10 years did not progress to having more severe restriction. Eighty‐one percent of the patients who had severe spinal restriction in 1980 were severely restricted within the first 10 years. Hips that were normal after 10 years of disease did not become diseased subsequently. This study suggests that a predictable pattern of ankylosing spondylitis emerges within the first 10 years of the disease.

OBJECTIVE Tosystematically evaluate ERG alterations in the multifocal tumor context by using whole-mount prostatectomy specimens of African American and Caucasian American patients matched for age, pathologic grade and stage. Oncogenic activation of the ETS-Related Gene (ERG) is the most common early genomic alteration in prostate cancer patients in Western countries. However, ERG alterations have not been systematically examined in African American patients with known higher risk of prostate cancer incidence and mortality. METHODS ERG oncoprotein expression was analyzed in 91 Caucasian American and 91 African American prostate cancer patients matched for age, Gleason score and pathologic stage. A unique aspect of this study was the evaluation of ERG in whole-mount prostatectomy sections, minimizing sampling bias and allowing the careful assessment of ERG in the multifocal tumor context of prostate cancer. RESULTS The frequency of ERG positive prostate tumors was significantly greater among Caucasian Americans vs. African Americans when assessed in all tumor foci (41.9% vs. 23.9%, p<0.0001). Markedly higher frequency of the ERG oncoprotein expression was noted between the index tumors of Caucasian Americans (63.3%) and African Americans (28.6%). Of note, in African American patients the higher grade index tumors were predominantly ERG negative. CONCLUSIONS ERG typing of prostate tumors establishes a major difference between the index tumors of Caucasian and African American patients. ERG negative index tumors may indicate less favorable outcome in African American patients. This study underscores that typing of prostate tumors for ERG may enhance our understanding of biological differences between the examined ethnic groups.

ObjectiveTo examine the effectiveness of green-Mediterranean (MED) diet, further restricted in red/processed meat, and enriched with green plants and polyphenols on non-alcoholic fatty liver disease (NAFLD), reflected by intrahepatic fat (IHF) loss.DesignFor the DIRECT-PLUS 18-month randomized clinical trial, we assigned 294 participants with abdominal obesity/dyslipidaemia into healthy dietary guidelines (HDG), MED and green-MED weight-loss diet groups, all accompanied by physical activity. Both isocaloric MED groups consumed 28 g/day walnuts (+440 mg/day polyphenols provided). The green-MED group further consumed green tea (3–4 cups/day) and Mankai (a Wolffia globosa aquatic plant strain; 100 g/day frozen cubes) green shake (+1240 mg/day total polyphenols provided). IHF% 18-month changes were quantified continuously by proton magnetic resonance spectroscopy (MRS).ResultsParticipants (age=51 years; 88% men; body mass index=31.3 kg/m2; median IHF%=6.6%; mean=10.2%; 62% with NAFLD) had 89.8% 18-month retention-rate, and 78% had eligible follow-up MRS. Overall, NAFLD prevalence declined to: 54.8% (HDG), 47.9% (MED) and 31.5% (green-MED), p=0.012 between groups. Despite similar moderate weight-loss in both MED groups, green-MED group achieved almost double IHF% loss (−38.9% proportionally), as compared with MED (−19.6% proportionally; p=0.035 weight loss adjusted) and HDG (−12.2% proportionally; p<0.001). After 18 months, both MED groups had significantly higher total plasma polyphenol levels versus HDG, with higher detection of Naringenin and 2-5-dihydroxybenzoic-acid in green-MED. Greater IHF% loss was independently associated with increased Mankai and walnuts intake, decreased red/processed meat consumption, improved serum folate and adipokines/lipids biomarkers, changes in microbiome composition (beta-diversity) and specific bacteria (p<0.05 for all).ConclusionThe new suggested strategy of green-Mediterranean diet, amplified with green plant-based proteins/polyphenols as Mankai, green tea, and walnuts, and restricted in red/processed meat can double IHF loss than other healthy nutritional strategies and reduce NAFLD in half.Trial registration numberNCT03020186.

Little is known about health variables and if these variables could increase the risk of injuries among adolescent elite athletes. The primary aim was to present overall data on self-perceived stress, nutrition intake, self-esteem, and sleep, as well as gender and age differences, on two occasions among adolescent elite athletes. A secondary aim was to study these health variables as potential risk factors on injury incidence. A questionnaire was e-mailed to 340 adolescent elite athletes on two occasions during a single school year: autumn semester and spring semester. The results show that during autumn semester, the recommended intake of fruits, vegetables, and fish was not met for 20%, 39%, and 43% of the adolescent elite athletes, respectively. The recommended amount of sleep during weekdays was not obtained by 19%. Multiple logistic regression showed that athletes sleeping more than 8 h of sleep during weekdays reduced the odds of injury with 61% (OR, 0.39; 95% CI, 0.16-0.99) and athletes reaching the recommended nutrition intake reduced the odds with 64% (OR, 0.36; 95% CI, 0.14-0.91). Our findings suggest that nutrition intake and sleep volume are of importance in understanding injury incidence.

Many risk factors for injury are presented in the literature, few of those are however consistent and the majority is associated with adult and not adolescent elite athletes. The aim was to identify risk factors for injury in adolescent elite athletes, by applying a biopsychosocial approach. A total of 496 adolescent elite athletes (age range 15-19), participating in 16 different sports, were monitored repeatedly over 52 weeks using a valid questionnaire about injuries, training exposure, sleep, stress, nutrition, and competence-based self-esteem. Univariate and multiple Cox regression analyses were used to calculate hazard ratios (HR) for risk factors for first reported injury. The main finding was that an increase in training load, training intensity, and at the same time decreasing the sleep volume resulted in a higher risk for injury compared to no change in these variables (HR 2.25, 95% CI, 1.46-3.45, P<.01), which was the strongest risk factor identified. In addition, an increase by one score of competence-based self-esteem increased the hazard for injury with 1.02 (HR 95% CI, 1.00-1.04, P=.01). Based on the multiple Cox regression analysis, an athlete having the identified risk factors (Risk Index, competence-based self-esteem), with an average competence-based self-esteem score, had more than a threefold increased risk for injury (HR 3.35), compared to an athlete with a low competence-based self-esteem and no change in sleep or training volume. Our findings confirm injury occurrence as a result of multiple risk factors interacting in complex ways.

A considerable number of elite adolescent athletes were injured weekly, resulting in serious consequences for sport participation, training, or performance (or a combination of these). Appropriately designed interventions to prevent knee and foot injuries will target both the greatest number of injuries and the injuries with the most serious consequences in elite adolescent athletes.

ObjectiveTo examine the joint associations of daily time spent in different intensities of physical activity, sedentary behaviour and sleep with all-cause mortality.MethodsFederated pooled analysis of six prospective cohorts with device-measured time spent in different intensities of physical activity, sedentary behaviour and sleep following a standardised compositional Cox regression analysis.Participants130 239 people from general population samples of adults (average age 54 years) from the UK, USA and Sweden.Main outcomeAll-cause mortality (follow-up 4.3–14.5 years).ResultsStudies using wrist and hip accelerometer provided statistically different results (I2=92.2%, Q-test p<0.001). There was no association between duration of sleep and all-cause mortality, HR=0.96 (95% CI 0.67 to 1.12). The proportion of time spent in moderate to vigorous physical activity was significantly associated with lower risk of all-cause mortality (HR=0.63 (95% CI 0.55 to 0.71) wrist; HR=0.93 (95% CI 0.87 to 0.98) hip). A significant association for the ratio of time spent in light physical activity and sedentary time was only found in hip accelerometer-based studies (HR=0.5, 95% CI 0.42 to 0.62). In studies based on hip accelerometer, the association between moderate to vigorous physical activity and mortality was modified by the balance of time spent in light physical activity and sedentary time.ConclusionThis federated analysis shows a joint dose–response association between the daily balance of time spent in physical activity of different intensities and sedentary behaviour with all-cause mortality, while sleep duration does not appear to be significant. The strongest association is with time spent in moderate to vigorous physical activity, but it is modified by the balance of time spent in light physical activity relative to sedentary behaviour.