Antibody-ligand interactions on a high-capacity staphylococcal protein A resin.

Staphylococcal protein-A affinity chromatography has been optimized for antibody purification, achieving a current capacity of up to 90 mg/ml in packed bed. The morphology of the particles, the number of antibodies bound per ligand and the spatial arrangement of the ligands were assessed by in-situ Small-angle X-ray scattering (SAXS) and scanning electron microscopy (SEM) combined with measurement of adsorption isotherms. We employed SAXS measurements to probe the nanoscale structure of the chromatographic resin. From scanning electron microcopy, the morphology and area of the beads were obtained. The adsorption isotherm revealed a bi-Langmuirian behavior where the association constant varied with the critical bulk concentration, indicating multilayer adsorption. Determining the antibody-ligand stoichiometry was crucial for understanding the adsorption mechanism, which was estimated to be 4 at lower concentrations and 4.5 at higher concentrations, suggestive of reversible protein-protein interactions. The same results were reached from the in-situ small angle X-ray scattering measurements. A stoichiometry of 6 cannot be achieved since the two protein A monomers are anchored to the stationary phase and thus sterically hindered. Normalization through ellipsoids facilitated SAXS analysis, enabling the determination of distances between ligands and antibody-ligand complexes. Density fluctuations were examined by subtracting the elliptical fit, providing insights into ligand density distribution. The dense ligand packing of TOYOPEARL® AF-rProtein A HC was confirmed, making further increases in ligand density impractical. Additionally, SAXS analysis revealed structural rearrangements of the antibody-ligand complex with increasing antibody surface load, suggesting reversible association of antibodies.

Loop diuretic discontinuation in chronic heart failure patients: A retrospective study.

INTRODUCTION AND OBJECTIVES: The use of loop diuretics is central in managing congestion in heart failure (HF), but their impact on prognosis remains unclear. In euvolemic patients, dose reduction is recommended, but there is no recommendation on their discontinuation. This study aims to assess the impact of loop diuretic discontinuation on the prognosis of outpatients with HF with reduced ejection fraction. METHODS: This retrospective cohort study collected data from medical records of patients followed in an outpatient HF clinic at a university hospital center. Patients were included if they had been on loop diuretics and these were discontinued. Demographic, clinical and laboratory data were collected, and number and type of congestive events during the one-year period after discontinuation were recorded. RESULTS: Among 265 patients on loop diuretics, almost half (129) discontinued them at some point. Patients had optimized medical therapy, low median age, low New York Heart Association class, low B-type natriuretic peptide values, normal blood pressure, controlled heart rate and kidney function within normal limits. Among 122 patients with one year of follow-up, 18 (14.8%) had a congestive event. Fifteen events (83.3%) were low-dose diuretic reinitiation at a scheduled visit. There were only three worsening heart failure events (2.5%) during the one-year period. A significant improvement in kidney function from discontinuation to the one-year follow-up appointment was also observed. CONCLUSIONS: In our cohort, loop diuretic discontinuation was possible and safe in a large proportion of patients. The results should be interpreted with caution and cannot be extrapolated to a broader population of HF patients.

Real-time reverse transcription recombinase polymerase amplification (RT-RPA) assay for detection of cassava brown streak viruses.

Cassava brown streak disease (CBSD) caused by Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) is the most economically important viral disease of cassava. As cassava is a vegetatively propagated crop, the development of rapid and sensitive diagnostics would aid in the identification of virus-free planting material and development of effective management strategies. In this study, a rapid, specific and sensitive real-time reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed for real-time detection of CBSV and UCBSV. The RT-RPA was able to detect as little as 2 pg/µl of purified RNA obtained from infected cassava leaves, a sensitivity equivalent to that obtained by quantitative real-time reverse transcription PCR (qRT-PCR), within 20 min at 37 °C. Further, the RT-RPA detected each target virus directly from crude leaf and stem extracts, avoiding the tedious and costly isolation of high-quality RNA. The developed RT-RPA assay provides a valuable diagnostic tool that can be adopted by cassava seed certification and virus resistance breeding programs to ensure distribution of virus-free cassava planting materials to farmers. This is the first report on the development and validation of crude sap-based RT-RPA assay for the detection of cassava brown streak viruses (UCBSV and CBSV) infection in cassava plants.

Cytogenomic Characterization of Transposable Elements and Satellite DNA in Passiflora L. Species.

The species Passiflora alata, P. cincinnata, and P. edulis have great economic value due to the use of their fruits for human consumption. In this study, we compared the repetitive genome fractions of these three species. The compositions of the repetitive DNA of these three species' genomes were analyzed using clustering and identification of the repetitive sequences with RepeatExplorer. It was found that repetitive DNA content represents 74.70%, 66.86%, and 62.24% of the genome of P. alata, P. edulis, and P. cincinnata, respectively. LTR Ty3/Gypsy retrotransposons represent the highest genome proportions in P. alata and P. edulis, while Ty1/Copia comprises the largest proportion of P. cincinnata genome. Chromosomal mapping by Fluorescent In Situ Hybridization (FISH) showed that LTR retrotransposons have a dispersed distribution along chromosomes. The subtelomeric region of chromosomes is where 145 bp satellite DNA is located, suggesting that these elements may play important roles in genome structure and organization in these species. In this work, we obtained the first global characterization of the composition of repetitive DNA in Passiflora, showing that an increase in genome size is related to an increase in repetitive DNA, which represents an important evolutionary route for these species.

Unravelling the molecular interactions behind the formation of PEG/PPG aqueous two-phase systems.

The understanding of molecular interactions that control phase separation in polymer/polymer aqueous two-phase systems (ATPS) has been a subject of debate up to this day. In light of this, we set out to investigate the molecular interactions occurring in ternary mixtures composed of polyethylene glycol (PEG600), polypropylene glycol (PPG400) and water. The ternary phase diagram was plotted at two temperatures (298 K and 323 K), revealing a transition from a type 0 to a type I diagram. Molecular dynamics (MD) simulations were performed to elucidate the polymer-polymer and polymer-water interactions occurring at different temperatures and water concentrations. COnductor-like Screening Model for Realistic Solvents (COSMO-RS) was used to assess the thermodynamic properties of the polymer-water binary mixtures and their correlation with ATPS formation. The MD simulations clearly demonstrate the effect of segregation/separation with increasing water content and temperature, highlighting a significant reduction in PPG-water interactions compared to PEG-water counterparts. Polymer-water interactions were identified as those controlling the phase separation mechanism, and the thermodynamic properties determined with COSMO-RS for the polymer-water binary systems further support this view.

Syringe irrigation in confluent canals: A sequential computational fluid dynamics assessment.

This study aims to assess the influence of root canal preparation, irrigation needle design and its placement depth in the irrigation flow of confluent canals during syringe irrigation. A mandibular molar presenting two confluent canals in its mesial root was sequentially prepared and scanned by micro-computed tomography after mechanical preparation up to ProTaper Next system sizes X2 (25/.06v), X3 (30/.07v) and X4 (40/.06v). In each of the root canal preparation models, a side-vented and an open-ended needle at 5, 3 and 2 mm from the working length were included, and irrigation flow was assessed by a validated computational fluid dynamics model. The results revealed that the irrigant flowed out of the confluent canals mainly through the canal that did not have the needle. Apical penetration and renewal of the irrigant were most efficiently achieved with the use of a 30G open-ended needle and a 30/.07v preparation.

Biophysical characterization of the tennis serve: A systematic scoping review with evidence gap map.

OBJECTIVES: We aimed to assess the available evidence on the biophysics of the tennis serve, mapping the populations, interventions, contexts and other relevant information to highlight what is already known and to identify gaps in the literature. DESIGN: Systematic scoping review with evidence gap map. METHODS: The protocol was designed according to PRISMA 2020, Prisma-ScR guidelines and the Cochrane Handbook. The searches were conducted on July 20, 2022 and updated on April 1, 2023, in PubMed, Scopus and Web of Science (core collection). The risk of bias assessment was performed using the Cochranes method for nonrandomized studies (RoBANS) and a narrative synthesis of the main findings was performed and supplemented with an evidence gap map. RESULTS: Most trials were found on serve kinematics and kinetics (95 %), analyzing only flat serves (84 and 72 %, respectively). Few trials focused on physiology (20 %; e.g. biomarkers), under-19, left or both-handed, female and intermediate beginner or starter players (29, 17, 8 and 7 %, respectively). We found a preponderance of low and unclear risk of bias (63 and 31 %, respectively) and only 7 % high, particularly, on the assessment of confounding variables. CONCLUSIONS: The current scoping review reveals a few trials on physiological rather than biomechanical variables, as well as the absence of the kick and slice serve, foot-back and foot-up serve, and left-handed, female, and young player analyses. We did not find systematic mistakes or limitations in the design, conduct, or analysis that would distort the results, since only 7 % presented a high risk of bias.

Acute Recovery after a Fatigue Protocol Using a Recovery Sports Legging: An Experimental Study.

Enhancing recovery is a fundamental component of high-performance sports training since it enables practitioners to potentiate physical performance and minimise the risk of injuries. Using a new sports legging embedded with an intelligent system for electrostimulation, localised heating and compression (completely embodied into the textile structures), we aimed to analyse acute recovery following a fatigue protocol. Surface electromyography- and torque-related variables were recorded on eight recreational athletes. A fatigue protocol conducted in an isokinetic dynamometer allowed us to examine isometric torque and consequent post-exercise acute recovery after using the sports legging. Regarding peak torque, no differences were found between post-fatigue and post-recovery assessments in any variable; however, pre-fatigue registered a 16% greater peak torque when compared with post-fatigue for localised heating and compression recovery methods. Our data are supported by recent meta-analyses indicating that individual recovery methods, such as localised heating, electrostimulation and compression, are not effective to recover from a fatiguing exercise. In fact, none of the recovery methods available through the sports legging tested was effective in acutely recovering the torque values produced isometrically.

Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the Rapid and Sensitive Detection of Yam mosaic virus.

Yam (Dioscorea spp.) productivity is constrained significantly by the lack of a formal seed system. Vegetative propagation, through tuber setts as 'seed' yams, encourages the recycling of virus-infected planting materials, contributing to high virus incidence and yield losses. Efforts are ongoing to increase the production of high-quality seed yams in a formal seed system to reduce virus-induced yield losses and enhance the crop's productivity and food security. Specific and sensitive diagnostic tests are imperative to prevent the multiplication of virus-infected materials contributing to a sustainable seed yam certification system. During routine indexing of yam accessions, discrepancies were observed between the results obtained from the reverse transcription loop-mediated isothermal amplification (RT-LAMP) test and those from reverse transcription polymerase chain reaction (RT-PCR); RT-LAMP failed to detect Yam mosaic virus (YMV) in some samples that tested positive by RT-PCR. This prompted the design of a new set of LAMP primers, YMV1-OPT primers. These primers detected as little as 0.1 fg/µL of purified RNA obtained from a YMV-infected plant, a sensitivity equivalent to that obtained with RT-PCR. RT-LAMP using YMV1-OPT primers is recommended for all future virus-indexing of seed yams for YMV, offering a rapid, sensitive, and cost-effective approach.

Bulk-Fill Resins versus Conventional Resins: An Umbrella Review.

Currently, composite resins have become the material of choice for the restoration of posterior teeth. Although bulk-fill resins represent a tempting alternative due to their lower complexity and faster use, some dentists are reluctant to use this material. The objective is to compare the performance of bulk-fill resins and conventional resins in direct restorations of posterior teeth based on the literature. The databases that were used to carry out the research were PubMed/MEDLINE, Embase, the Cochrane Library and the WOS. This umbrella literature review complies with PRISMA standards and assesses the quality of studies using the AMSTAR 2 tool. With the application of the criteria of the AMSTAR 2 tool, the reviews were considered low to moderate. The overall meta-analysis, although without statistical significance, favours mostly the use of conventional resin, as it is about five times more likely to obtain a favourable result than bulk-fill resin. Bulk-fill resins result in a simplification of the clinical process of posterior direct restorations, which is an advantage. The performance in terms of several properties of bulk-fill resins and conventional resins showed that they present similar behaviour.

Data fusion and multivariate analysis for food authenticity analysis.

A mid-level data fusion coupled with multivariate analysis approach is applied to dual-platform mass spectrometry data sets using Rapid Evaporative Ionization Mass Spectrometry and Inductively Coupled Plasma Mass Spectrometry to determine the correct classification of salmon origin and production methods. Salmon (n = 522) from five different regions and two production methods are used in the study. The method achieves a cross-validation classification accuracy of 100% and all test samples (n = 17) have their origins correctly determined, which is not possible with single-platform methods. Eighteen robust lipid markers and nine elemental markers are found, which provide robust evidence of the provenance of the salmon. Thus, we demonstrate that our mid-level data fusion - multivariate analysis strategy greatly improves the ability to correctly identify the geographical origin and production method of salmon, and this innovative approach can be applied to many other food authenticity applications.

Physical Fitness Profile of High-Level Female Portuguese Handball Players.

We characterized the physical and physiological profiles of high-level female Portuguese handball players and examined the relationships between their anthropometric characteristics, general motor performance and cardiopulmonary fitness. Twenty-four high-level female handball players with an average age of 23.6 ± 5.5 years, height of 173.6 ± 5.1 cm and body mass of 72.6 ± 9.1 kg volunteered to participate. A Pearson correlation test was used to assess the relationship between variables. Direct relationships were observed between the players' height and arm span (r = 0.741), as well as between their squat jump and countermovement jump performances with regard to body mass (r = 0.448 and 0.496, respectively). The 9 m jump shot has a large relationship with the 7 m standing throw (r = 0.786) and between left hand dynamometry and body mass index (r = 0.595). The 30 m sprint has a relationship with the 7 m standing throw (r = -0.526) and the 9 m jump throw (r = -0.551). Oxygen uptake has a relationship with the players' height (r = -0.482) and time limit (r = 0.513), while the fitness index has a relation to the players' height (r = -0.488) and arm span (r = -0.422). Our results should be considered when using physical testing to plan optimal physical training regimens in elite team handball.

The path towards type V deep eutectic solvents: inductive effects and steric hindrance in the system tert-butanol + perfluoro tert-butanol.

The solid-liquid phase behaviour of two tertiary alcohols, perfluoro-tert-butanol and tert-butanol, was studied here using experimental (ITC, DSC and density measurements) and theoretical (MD simulations) approaches. The phase diagram of the binary mixture reveals highly negative deviations from ideality at low concentrations, as well as the formation of co-crystals and is characterized by two eutectic points, a congruent melting point and a peritectic reaction corresponding to TBH : TBF stoichiometries of 2 : 1 and 1 : 1 respectively. Excess molar enthalpies and volumes were calculated, showing negative and positive deviations from ideality, respectively. The effect of acidity, stereochemical hindrance and phobic effects and how they affect intermolecular interactions in these binary mixtures is discussed, with the aim of designing and fine-tuning type V deep eutectic solvents. The results showed that the fluorination of tertiary alcohols can be used for the tuning of the mixing properties and solid-liquid phase diagrams.

Unified directional parabolic-accurate lattice Boltzmann boundary schemes for grid-rotated narrow gaps and curved walls in creeping and inertial fluid flows.

The goal of this work is to advance the characteristics of existing lattice Boltzmann Dirichlet velocity boundary schemes in terms of the accuracy, locality, stability, and mass conservation for arbitrarily grid-inclined straight walls, curved surfaces, and narrow fluid gaps, for both creeping and inertial flow regimes. We reach this objective with two infinite-member boundary classes: (1) the single-node "Linear Plus" (LI^{+}) and (2) the two-node "Extended Multireflection" (EMR). The LI^{+} unifies all directional rules relying on the linear combinations of up to three pre- or postcollision populations, including their "ghost-node" interpolations and adjustable nonequilibrium approximations. On this basis, we propose three groups of LI^{+} nonequilibrium local corrections: (1) the LI_{1}^{+} is parametrized, meaning that its steady-state solution is physically consistent: the momentum accuracy is viscosity-independent in Stokes flow, and it is fixed by the Reynolds number (Re) in inertial flow; (2) the LI_{3}^{+} is parametrized, exact for arbitrary grid-rotated Poiseuille force-driven Stokes flow and thus most accurate in porous flow; and (3) the LI_{4}^{+} is parametrized, exact for pressure and inertial term gradients, and hence advantageous in very narrow porous gaps and at higher Reynolds range. The directional, two-relaxation-time collision operator plays a crucial role for all these features, but also for efficiency and robustness of the boundary schemes due to a proposed nonequilibrium linear stability criterion which reliably delineates their suitable coefficients and relaxation space. Our methodology allows one to improve any directional rule for Stokes or Navier-Stokes accuracy, but their parametrization is not guaranteed. In this context, the parametrized two-node EMR class enlarges the single-node schemes to match exactness in a grid-rotated linear Couette flow modeled with an equilibrium distribution designed for the Navier-Stokes equation (NSE). However, exactness of a grid-rotated Poiseuille NSE flow requires us to perform (1) the modification of the standard NSE term for exact bulk solvability and (2) the EMR extension towards the third neighbor node. A unique relaxation and equilibrium exact configuration for grid-rotated Poiseuille NSE flow allows us to classify the Galilean invariance characteristics of the boundary schemes without any bulk interference; in turn, its truncated solution suggests how, when increasing the Reynolds number, to avoid a deterioration of the mass-leakage rate and momentum accuracy due to a specific Reynolds scaling of the kinetic relaxation collision rate. The optimal schemes and strategies for creeping and inertial regimes are then singled out through a series of numerical tests, such as grid-rotated channels and rotated Couette flow with wall-normal injection, cylindrical porous array, and Couette flow between concentric cylinders, also comparing them against circular-shape fitted FEM solutions.

Viruses of Yams (Dioscorea spp.): Current Gaps in Knowledge and Future Research Directions to Improve Disease Management.

Viruses are a major constraint for yam production worldwide. They hamper the conservation, movement, and exchange of yam germplasm and are a threat to food security in tropical and subtropical areas of Africa and the Pacific where yam is a staple food and a source of income. However, the biology and impact of yam viruses remains largely unknown. This review summarizes current knowledge on yam viruses and emphasizes gaps that exist in the knowledge of the biology of these viruses, their diagnosis, and their impact on production. It provides essential information to inform the implementation of more effective virus control strategies.

Karyotyping of commercial cultivars of melon (Cucumis melo L.).

BACKGROUND: This study on cultivars of melon (Cucumis melo L.) marketed in Brazil was conducted to obtain information to be used in breeding programs of this species. Little is known about the karyotype variability among C. melo L. cultivars targeted at the consumer market. The objective of the present study was to verify the karyotype variability in eight commercial melon cultivars used in the Brazilian market. METHODS AND RESULTS: Slides were stained with 2% Giemsa and assembled with Neomount to perform chromosomal morphometry. GC-rich heterochromatin was observed by CMA3/DAPI staining. 5 S rDNA, centromeric satellite DNA (SatDNA), and telomeric sites were visualized using fluorescence in situ hybridization. All images were captured on an Olympus BX41 microscope equipped with a 5 M Olympus DP25 digital camera and DP2-BSW software. The cultivars showed symmetrical karyotypes with significant differences in total chromosome length and average chromosome size. Heterochromatic CMA3+ blocks were observed in terminal regions related to satellites (secondary constrictions), as well as in centromeric and pericentromeric regions. A single chromosomal pair of 5 S rDNA sites was observed in all cultivars, but at distinct locations. Centromeric satellite sequences, tested for the first time in melon, revealed only centromeric sites. Telomeric sites were observed in all the chromosomes of the cultivars. CONCLUSIONS: Karyotype variation was observed in cultivars of melon, which were analyzed for chromosomal morphology and localization of GC-rich heterochromatin, as well centromeric SatDNA, rDNA, and telomeric chromosomal markers. Hence, these cultivars can be used in future breeding programs.

Lower Limbs Wearable Sports Garments for Muscle Recovery: An Umbrella Review.

This review aims to understand the different technologies incorporated into lower limbs wearable smart garments and their impact on post-exercise recovery. Electronic searches were conducted in the PubMed, Web of Science, and Cochrane electronic databases. Eligibility criteria considered meta-analyses that examined the effects of wearable smart garments on physical fitness in healthy male and female adults. Seven meta-analyses were considered in the current umbrella review, indicating small effects on delayed-onset muscle soreness ([0.40-0.43]), rate of perceived exertion (0.20), proprioception (0.49), anaerobic performance (0.27), and sprints ([0.21-0.37]). The included meta-analyses also indicated wearable smart garments have trivial to large effects on muscle strength and power ([0.14-1.63]), creatine kinase ([0.02-0.44]), lactate dehydrogenase (0.52), muscle swelling (0.73), lactate (0.98) and aerobic pathway (0.24), and endurance (0.37), aerobic performance (0.60), and running performance ([0.06-6.10]). Wearing wearable smart garments did not alter the rate of perceived exertion and had a small effect on delayed-onset muscle soreness. Well-fitting wearable smart garments improve comfort and kinesthesia and proprioception and allow a reduction in strength loss and muscle damage after training and power performance following resistance training or eccentric exercise.

Experimental validation of a computational fluid dynamics model using micro-particle image velocimetry of the irrigation flow in confluent canals.

AIM: This study aimed to experimentally validate a computational fluid dynamics (CFD) model, using micro-particle image velocimetry (micro-PIV) measurements of the irrigation flow velocity field developed in confluent canals during irrigation with a side-vented needle. METHODOLOGY: A microchip with confluent canals, manufactured in polydimethylsiloxane was used in a micro-PIV analysis of the irrigation flow using a side-vented needle placed 3 mm from the end of the confluence of the canals. Velocity fields and profiles were recorded for flow rates of 0.017 and 0.1 ml/s and compared with those predicted in CFD numerical simulations (using a finite volume commercial code - FLUENT) for both laminar and turbulent regimes. RESULTS: The overall flow pattern, isovelocity and vector maps as well as velocity profiles showed a close agreement between the micro-PIV experimental and CFD predicted data. No relevant differences were observed between the results obtained with the laminar and turbulent flow models used. CONCLUSIONS: Results showed that the laminar CFD modelling is reliable to predict the flow in similar domains.