Trimethylation profile of histones H3 lysine 4 and 9 in late preantral and early antral caprine follicles grown in vivo versus in vitro in the presence of anethole.

This study assessed the histones methylation profile (H3K4me3 and H3K9me3) in late preantral (PA) and early antral (EA) caprine follicles grown in vivo and in vitro, and the anethole effect during in vitro culture of PA follicles. Uncultured in vivo-grown follicles (PA, n = 64; EA, n = 73) were used as controls to assess the methylation profile and genes' expression related to apoptosis cascade (BAX, proapoptotic; BCL2, antiapoptotic), steroidogenesis (CYP17, CYP19A1), and demethylation (KDM1AX1, KDM1AX2, KDM3A). The isolated PA follicles (n = 174) were cultured in vitro for 6 days in α-MEM+ in either absence (control) or presence of anethole. After culture, EA follicles were evaluated for methylation, mRNA abundance, and morphometry. Follicle diameter increased after culture, regardless of treatment. The methylation profile and the mRNA abundance were similar between in vivo-grown PA and EA follicles. Anethole treatment led to higher H3K4me3 fluorescence intensity in EA follicles. The mRNA abundances of BAX, CYP17, and CYP19A1 were higher, and BCL2 and KDM3A were lower in in vitro-grown EA follicles than in vivo-grown follicles. In conclusion, in vitro follicle culture affected H3K4me3 fluorescence intensity, mRNA abundance of apoptotic genes, and steroidogenic and demethylase enzymes compared with in vivo-grown follicles.

Exercise training counteracts the cardiac metabolic remodelling induced by experimental pulmonary arterial hypertension.

Exercise training provides several cardiovascular benefits in both physiological and pathological conditions; however, its use as a therapeutic tool for pulmonary arterial hypertension (PAH) has been poorly explored. This study aimed to extend the comprehension of the cardioprotective effects of exercise training in the set of PAH focusing on the metabolic changes promoted by exercise in the right ventricle (RV). The monocrotaline animal model of PAH was used and male Wistar rats were submitted to two weeks of treadmill exercise training (5 days/week, 60 min/day, 25 m/min) following disease establishment. Trained rats showed an improved diastolic function (lower end-diastolic pressure and tau) despite the presence of cardiac overload (increased peak systolic pressure, end-diastolic pressure and arterial elastance). This enhanced hemodynamic response was paralleled by an increased uptake of glucose to cardiomyocytes through glucose transporter type 4 (GLUT4) followed by increased lactate dehydrogenase (LDH) activity. Exercise did not reverse the decrease of fatty acid oxidation related to PAH but increased the content of the transcription factors peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and peroxisome proliferator-activated receptor gamma (PPAR-γ). Two weeks of exercise did not modulate the changes in amino acid metabolism secondary to PAH. Our work suggests that continuous aerobic exercise of moderate intensity, despite its short-term duration and application in a late stage of the disease, supports the RV response to PAH by promoting a shift in the cardiac metabolic phenotype.

The Selective Late Sodium Current Inhibitor Eleclazine, Unlike Amiodarone, Does Not Alter Defibrillation Threshold or Dominant Frequency of Ventricular Fibrillation.

INTRODUCTION: We examined the effects of the selective late INa inhibitor eleclazine on the 50% probability of successful defibrillation (DFT50) before and after administration of amiodarone to determine its suitability for use in patients with implantable cardioverter defibrillators (ICDs). METHODS AND RESULTS: In 20 anesthetized pigs, transvenous active-fixation cardiac defibrillation leads were fluoroscopically positioned into right ventricular apex through jugular vein. ICDs were implanted subcutaneously. Dominant frequency of ventricular fibrillation was analyzed by fast Fourier transform. The measurements were made before drug administration (control), and at 40 minutes after vehicle, eleclazine (2 mg/kg, i.v., bolus over 15 minutes), or subsequent/single amiodarone administration (10 mg/kg, i.v., bolus over 10 minutes). Eleclazine did not alter DFT50, dominant frequency, heart rate, or mean arterial pressure (MAP). Subsequent amiodarone increased DFT50 (P = 0.006), decreased dominant frequency (P = 0.022), and reduced heart rate (P = 0.031) with no change in MAP. Amiodarone alone increased DFT50 (P = 0.005; NS compared to following eleclazine) and decreased dominant frequency (P = 0.003; NS compared to following eleclazine). CONCLUSION: Selective late INa inhibition with eleclazine does not alter DFT50 or dominant frequency of ventricular fibrillation when administered alone or in combination with amiodarone. Accordingly, eleclazine would not be anticipated to affect the margin of defibrillation safety in patients with ICDs.

Impact of sludge retention time on MBR fouling: role of extracellular polymeric substances determined through membrane autopsy.

The impact of sludge retention time (SRT) on the biofouling of a membrane bioreactor (MBR) by extracellular polymeric substances (EPS) was investigated. The MBR was operated at 60 and 20 d SRT. The gel layer (recovered through optimized membrane autopsy methods) and the cake layer were analyzed for their content and profile of EPS proteins and polysaccharides. The change to a shorter SRT led to decreased membrane filterability, concomitant with a higher expression of EPS proteins in the cake layer, which were identified as being mainly related with biosynthesis and stress functions. The gel layer was more substantial in internal fibers, with polysaccharides being the major component in this layer. With the decrease in SRT (and filterability decrease), the overall polysaccharide content and sugar variety increased. In conclusion, SRT impacted not only on the quantity but also the composition of EPS molecules, and both were shown to be important in biofouling.

Impact of sludge retention time on the fine composition of the microbial community and extracellular polymeric substances in a membrane bioreactor.

Membrane bioreactors (MBRs) are an advanced technology for wastewater treatment whose wide application has been hindered by rapid fouling of the membranes. MBRs can be operated with long sludge retention time (SRT), a crucial parameter impacting microbial selection in the reactor. This also affects filtration performance, since a major fouling agent are the extracellular polymeric substances (EPS). In this study, the impact of the SRT on the ecophysiology of the MBRs and, consequently, on membrane fouling was evaluated. A MBR was operated under a SRT of 60 days followed by a SRT of 20 days. A comprehensive analysis of the microbial community structure and EPS proteins and polysaccharide profiles of the mixed liquor and cake layer was carried out throughout both operation periods. The results of this study showed that the imposition of a shorter SRT led to a shift in the dominant bacterial populations. The mixed liquor and cake layer communities were very different, with Actinomycetales order standing out in the cake layer at SRT of 20 days. Overall, higher EPS concentrations (particularly proteins) were found at this SRT. Furthermore, EPS profiles were clearly affected by the SRT: it was possible to correlate a group of soluble EPS proteins with the SRT of 60 days, and a lower sludge age led to a lower diversity of polysaccharide sugar monomers, with an increase of glucose and galactose in the cake layer. This study improves our knowledge regarding the molecular reasons for fouling, which may contribute to improve MBR design and operation.

Combined actions of ivabradine and ranolazine reduce ventricular rate during atrial fibrillation.

INTRODUCTION: Ventricular rate during atrial fibrillation (AF) can be reduced by slowing atrioventricular (AV) node conduction and/or by decreasing dominant frequency of AF. We investigated whether combined administration of ivabradine and ranolazine reduces ventricular rate during AF. METHODS AND RESULTS: Ivabradine (maximum clinical dose, 0.25 mg/kg, and 0.10 mg/kg, i.v.) and ranolazine (2.4 mg/kg, i.v., bolus followed by 0.135 mg/kg/min) were studied in an anesthetized pig (N = 16) model of AF. Combined administration of 0.25 mg/kg ivabradine with ranolazine reduced ventricular rate during AF by 51.9 ± 9.7 beats/min (23%, P = 0.017) and dominant frequency of AF by 2.8 ± 0.5 Hz (32%, P = 0.005). It increased PR (P = 0.0002, P = 0.0007) and A-H intervals (P = 0.047, P = 0.002) during pacing at 130 and 180 beats/min, respectively, to a greater degree than additive effects of single agents. Combined administration of 0.1 mg/kg ivabradine with ranolazine exceeded additive effects of single agents on A-H intervals and dominant frequency of AF. Moreover, ranolazine potentiated low-dose ivabradine's reduction in ventricular rate, as combined administration more than doubled effects of the higher ivabradine dose alone and was similar to the combination with the higher dose. Neither drug nor their combination affected contractility (left ventricular [LV] dP/dt), QT or His-ventricular (H-V) intervals, or mean arterial pressure during sinus rhythm or AF. CONCLUSION: Combined administration of ivabradine and ranolazine at clinically safe levels decreases ventricular rate during AF by reducing AV node conduction and AF dominant frequency without QT prolongation or depression in contractility. Targeting these actions offers intrinsic advantages over conventional nodal agents, which can reduce contractility.

Draft genomes of seven isolates from Danish wastewater facilities belonging to Pseudomonas, Bacillus, Pseudochrobactrum, Brevundimonas, and Pandoraea.

We report here seven draft genomes of bacterial strains from two Danish wastewater facilities, two of which might be characterized as a new group within the Pseudomonas and Pseudochrobactrum genera, respectively. These genomes will provide useful references for understanding bacterial interactions and horizontal gene transfer within bacterial communities.

Chemiluminometric evaluation of melatonin and selected melatonin precursors' interaction with reactive oxygen and nitrogen species.

Melatonin is a hormone, a derivative of tryptophan, that possesses a potent scavenging capacity for the most reactive and dangerous free radicals, being an important protection against oxidative stress. In this work, an automated flow-based procedure for assessment of melatonin, tryptophan, and 5-hydroxytryptophan scavenging capacity was developed. The presented methodology involved a multi-pumping flow system and exploited the ability of selected compounds to inhibit the chemiluminescence reaction of luminol with hydrogen peroxide, hydroxyl radical, and peroxynitrite anion. The system was based on the use of several solenoid actuated micro-pumps as the only active components of the flow manifold. This enabled the reproducible insertion and efficient mixing of very low volumes of sample and reagents as well as the transportation of the sample zone toward detection for monitoring the chemiluminometric response. Furthermore, the high versatility of the proposed multi-pumping flow system allowed the implementation of distinct reactions for the in-line generation of the different reactive species assayed without requiring physical reconfiguration. The results obtained demonstrated that 5-hydroxytryptophan is the most potent scavenger, followed by melatonin and tryptophan. The developed multi-pumping flow system exhibited good measurement precision (relative standard deviations typically <2%, n=10), low operational costs, and low reagent consumption.

Step-by-step strategy for protein enrichment and proteome characterisation of extracellular polymeric substances in wastewater treatment systems.

Extracellular polymeric substances (EPS) are keys in biomass aggregation and settleability in wastewater treatment systems. In membrane bioreactors (MBR), EPS are an important factor as they are considered to be largely responsible for membrane fouling. Proteins were shown to be the major component of EPS produced by activated sludge and to be correlated with the properties of the sludge, like settling, hydrophobicity and cell aggregation. Previous EPS proteomic studies of activated sludge revealed several problems, like the interference of other EPS molecules in protein analysis. In this study, a successful strategy was outlined to identify the proteins from soluble and bound EPS extracted from activated sludge of a lab-scale MBR. EPS samples were first subjected to pre-concentration through lyophilisation, centrifugal ultrafiltration or concentration with a dialysis membrane coated by a highly absorbent powder of polyacrylate-polyalcohol, preceded or not by a dialysis step. The highest protein concentration factors were achieved with the highly absorbent powder method without previous dialysis step. Four protein precipitation methods were then tested: acetone, trichloroacetic acid (TCA), perchloric acid and a commercial kit. Protein profiles were compared in 4-12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis gels. Both acetone and TCA should be applied for the highest coverage for soluble EPS proteins, whereas TCA was the best method for bound EPS proteins. All visible bands of selected profiles were subjected to mass spectrometry analysis. A high number of proteins (25-32 for soluble EPS and 17 for bound EPS) were identified. As a conclusion of this study, a workflow is proposed for the successful proteome characterisation of soluble and bound EPS from activated sludge samples.

Microbial population analysis of nutrient removal-related organisms in membrane bioreactors.

Membrane bioreactors (MBR) are an important and increasingly implemented wastewater treatment technology, which are operated at low food to microorganism ratios (F/M) and retain slow-growing organisms. Enhanced biological phosphorus removal (EBPR)-related organisms grow slower than ordinary heterotrophs, but have never been studied in detail in MBRs. This study presents a comprehensive analysis of the microorganisms involved in EBPR in pilot- and full-scale MBRs, using fluorescence in situ hybridization (FISH), as well as an overall assessment of other relevant microbial groups. The results showed that polyphosphate accumulating organisms (PAOs) were present at similar levels in all studied MBRs (10% ± 6%), even those without a defined anaerobic zone. Glycogen accumulating organisms were also detected, although rarely. The FISH results correlated well with the observed P removal performance of each plant. The results from this study suggest that a defined anaerobic zone is not necessarily required for putative PAO growth in MBRs, since polyphosphate storage may provide a selective advantage in fulfilling cell maintenance requirements in substrate-limited conditions (low F/M).

Front crawl swimming coordination: a systematic review.

Several constraints, including environmental (e.g., aquatic resistance, temperature and viscosity), organismic (e.g., anthropometry, buoyancy) and task-related (e.g., imposed swim speed or stroke rate) impact motor coordination and swimming performance. As motor coordination requires structurally organising intra- and inter-limb coupling, the purpose of this review was to update the literature concerning coordination between the upper-limbs in front crawl swimming. We focused on the effects of biomechanical, physiological, and personal (gender, skill level, and age) factors on coordination and performance. In fact, it could be highlighted that upper-limbs coordination varies with organismic, task and environmental constraints, resulting in several available motor solutions that should be adopted according to how each swimmer deals with occurring constraints. As such, there is no ideal or optimal coordination pattern that youth, learners and less-skilled swimmers should imitate.

The Effect of a Coordinative Training in Young Swimmers' Performance.

This study investigated the effects of a coordinative in-water training. Total 26 young swimmers (16 boys) were divided in a training group (that performed two sets of 6 × 25-m front crawl, with manipulated speed and stroke frequency, two/week for eight weeks) and a control group. At the beginning and end of the training period, swimmers performed 50-m front crawl sprints recorded by seven land and six underwater Qualisys cameras. A linear mixed model regression was applied to investigate the training effects adjusted for sex. Differences between sex were registered in terms of speed, stroke length, and stroke index, highlighting that an adjustment for sex should be made in the subsequent analysis. Between moments, differences were noticed in coordinative variables (higher time spent in anti-phase and push, and lower out-of-phase and recovery for training group) and differences between sex were noticed in performance (stroke length and stroke index). Interactions (group * time) were found for the continuous relative phase, speed, stroke length, and stroke index. The sessions exerted a greater (indirect) influence on performance than on coordinative variables, thus, more sessions may be needed for a better understanding of coordinative changes since our swimmers, although not experts, are no longer in the early learning stages.

Technical and economic evaluation of the integration of membrane bioreactor and air-stripping/absorption processes in the treatment of landfill leachate.

A membrane bioreactor inoculated with commercial baker's yeast (Saccharomyces cerevisiae) (MBRy) integrated to an air-stripping/absorption (AS/AB) as pre-treatment (aiming ammonia recovery) or a post-treatment (polishment step) was assessed for the landfill leachate treatment. The effect of chemical oxygen demand (COD) and nitrogen (N) ratio (C:N) on the performance of the MBRy was also investigated. At high COD/N ratio, high organic matter removal in terms of COD (71 ± 4%) and ammonia removal (97 ± 3%) was observed. Lower COD/N ratio favored yeast growth in the mixed liquor even under adverse conditions. The results of ammonia removal and recovery, and economic analysis demonstrated that the best way to integrate the AS/BS processes is as pre-treatment of MBRy. The ammonia concentration in the AS/AB process feed was a key factor to achieve the market specification. Although pH and temperature adjustment were adequate to promote ammonia removal/recovery, the AS operation at high temperatures showed the highest ammonia removal rate (99%). Therefore, the integration of AS/AB with MBRy allows obtaining a permeate with a final concentration of 2902 ± 374 mg L-1 of COD and 9 ± 7.5 mg L-1 of ammonia. Although it was possible to reach the Brazilian discharge standard for ammonia (20 mg L-1), it was not possible to reach the standard for COD, where the remaining fraction is recalcitrant organic matter, requiring the integration of a physico-chemical process. It should be noted that the proposed route allowed recovery 7 kg of ammonia per m3 of treated leachate.

The Relationship between Health Perception and Health Predictors among the Elderly across European Countries.

This study aimed to investigate the relationship between health perception and health predictors among the elderly. In this study, 376 older adults from four different countries (Hungary, n = 86; Italy, n = 133; Portugal, n = 95; and Spain, n = 62) were analyzed. All subjects completed the EQ-5D-5L to assess their quality-adjusted life years and were assessed in handgrip (HG) and in Timed Up and Go (TUG) tests. A three-way MANOVA was conducted to analyze the groups based on their age, sex, and country. The interaction effects in all included variables were also considered. The Bonferroni test was also executed as a post hoc test. Any interaction results were noticed. Regarding age, lower perceived quality of life scores and higher TUG results were registered in the oldest group, and greater values of left and right HG results were registered in the second-oldest group. Males showed greater left and right HG values than women. Spain showed lower perceived quality of life scores. Portugal and Italy showed greater HG left values, while Portugal had better HG right values. Hungary produced the greatest TUG scores. Quality of life is dependent on the subject's age and physical fitness, as increasing age was associated with decreased values of HG and TUG. Only strength was different between sexes.

Analyzing the Seasonal Changes and Relationships in Training Load and Wellness in Elite Volleyball Players

PURPOSE: The purposes of this study were to (1) analyze the variations of acute and chronic training load and well-being measures during 3 periods of the season (early, mid, and end) and (2) test the associations between weekly training load and well-being measures during different periods of the season. METHODS: Thirteen professional volleyball players from a team competing in the Portuguese Volleyball First Division (age 31.0 [5.0] y) were monitored during an entire season. Weekly acute (wAL) and chronic load (wCL), acute to chronic workload ratio (wACWL), and training monotony (wTM) were calculated during all weeks of the season. The weekly values of muscle soreness (wDOMS), stress (wStress), fatigue (wFatigue), sleep (wSleep), and Hooper index (wHI) were also obtained across the season. RESULTS: The midseason had meaningfully low values of wAL (-26.9%; effect size [ES]: -1.12) and wCL (-28.0%; ES: -2.81), and greater values of wACWL (+38.9%; ES: 2.81) compared with early season. The wCL (+10.6%; ES: 0.99), wStress (44.6%; ES: 0.87), and wHI (29.0%; ES: 0.62) were meaningfully greater during the end of season than in midseason. Overall, wAL presented very large correlations with wDOMS (r = .80), wSleep (r = .72), and wFatigue (r = .82). CONCLUSIONS: The results of this study suggest that the load was meaningfully higher during early season; however, stress was higher during the final stages of the season. Overall, it was also found that the acute load is more highly correlated with well-being status and its variations than chronic load or training monotony.

Integrated Analysis of Young Swimmers' Sprint Performance.

To analyze young swimmers' performance regarding sex and skill level, 23 boys and 26 girls (15.7 ± 0.8 and 14.5 ± 0.8 years old, respectively) were assessed for anthropometry, flexibility, strength, drag, coordination, and biomechanical variables. During a 50-m maximal front-crawl bout, seven aerial and six underwater Qualisys cameras assessed kinematics, and a load cell was used to measure drag (Tedea, United Kingdom) and tethered swimming force. A multivariate analysis of variance test (p < .05) enabled us to observe differences between skill levels in speed, stroke frequency, stroke index, and intracyclic velocity variations, but most relevant differences were noticed when comparing sexes, particularly for anthropometrics, shoulder flexibility, speed, stroke frequency, stroke length, drag, mechanical power, power per stroke, and maximal and mean force. Considering the included variables, only male swimmers' performance could be predicted through multiple linear regression, with stroke index, left shoulder flexion, and intracycle velocity variations showing great importance in achieving better results.

Task Constraints and Coordination Flexibility in Young Swimmers.

This study aimed to examine young swimmers' behavioral flexibility when facing different task constraints, such as swimming speed and stroke frequency. Eighteen (five boys and 13 girls) 13- to 15-year-old swimmers performed a 15 × 50-m front crawl with five trials, at 100%, 90%, and 70% each of their 50 m maximal swimming speed and randomly at 90%, 95%, 100%, 105%, and 110% of their preferred stroke frequency. Seven aerial and six underwater cameras were used to assess kinematics (one cycle), with upper-limb coordination computed through a continuous relative phase and index of coordination. A cluster analysis identified six patterns of coordination used by swimmers when facing various speed and stroke frequency constraints. The patterns' nature and the way the swimmers shifted between them are more important than getting the highest number of patterns (range of repertoire), that is, a change in the motor pattern in order to adapt correctly is more important than being able to execute a great number of patterns.

In-Depth Evaluation of Data Collected During a Continuous Pharmaceutical Manufacturing Process: A Multivariate Statistical Process Monitoring Approach.

The present work presents an in-depth evaluation of continuously collected data during a twin-screw granulation and drying process performed on a continuous manufacturing line. During operation, the continuous line logs 49 univariate process variables, hence generating a large amount of data. Three identical 5-h continuous manufacturing runs were performed. Multivariate data analysis tools, more specifically latent variable modeling tools such as principal component analysis, were used to extract information from the generated data sets unveiling process trends and drifts. Furthermore, a statistical process monitoring strategy is presented. The approach is based on the application of multivariate statistical process monitoring to model the variables that remain around a steady state.

Bioaugmentation of membrane bioreactor with Achromobacter denitrificans strain PR1 for enhanced sulfamethoxazole removal in wastewater.

Achromobacter denitrificans strain PR1, previously found to harbour specific degradation pathways with high sulfamethoxazole (SMX) degradation rates, was bioaugmented into laboratory-scale membrane bioreactors (MBRs) operated under aerobic conditions to treat SMX-containing real domestic wastewater. Different hydraulic retention times (HRTs), which is related to reaction time and loading rates, were considered and found to affect the SMX removal efficiency. The availability of primary substrates was important in both bioaugmented and non-bioaugmented activated sludge (AS) for cometabolism of SMX. High HRT (24 h) resulted in low food to microorganism ratio (F/M) and low SMX removal, due to substrate limitation. Decrease in HRT from 24 h to 12 h, 6 h and finally 4 h led to gradual increases in primary substrates availability, e.g. organic compounds and ammonia, resulted in increased SMX removal efficiency and degradation rate, and is more favorable for high-rate wastewater treatment processes. After inoculation into the MBRs, the bioaugmentation strain was sustained in the reactor for a maximum of 31 days even though a significant decrease in abundance was observed. The bioaugmented MBRs showed enhanced SMX removal, especially under SMX shock loads compared to the control MBRs. The results of this study indicate that re-inoculation is required regularly after a period of time to maintain the removal efficiency of the target compound.

Statistical process control of cocrystallization processes: A comparison between OPLS and PLS.

Orthogonal partial least squares regression (OPLS) is being increasingly adopted as an alternative to partial least squares (PLS) regression due to the better generalization that can be achieved. Particularly in multivariate batch statistical process control (BSPC), the use of OPLS for estimating nominal trajectories is advantageous. In OPLS, the nominal process trajectories are expected to be captured in a single predictive principal component while uncorrelated variations are filtered out to orthogonal principal components. In theory, OPLS will yield a better estimation of the Hotelling's T2 statistic and corresponding control limits thus lowering the number of false positives and false negatives when assessing the process disturbances. Although OPLS advantages have been demonstrated in the context of regression, its use on BSPC was seldom reported. This study proposes an OPLS-based approach for BSPC of a cocrystallization process between hydrochlorothiazide and p-aminobenzoic acid monitored on-line with near infrared spectroscopy and compares the fault detection performance with the same approach based on PLS. A series of cocrystallization batches with imposed disturbances were used to test the ability to detect abnormal situations by OPLS and PLS-based BSPC methods. Results demonstrated that OPLS was generally superior in terms of sensibility and specificity in most situations. In some abnormal batches, it was found that the imposed disturbances were only detected with OPLS.