High-field ex vivo and in vivo two-dimensional nuclear magnetic resonance spectroscopy in murine brain: Resolving and exploring the molecular environment.

The structural and chemical complexities within the brain pose a challenge that few noninvasive techniques can tackle with the dexterity of nuclear magnetic resonance (NMR) spectroscopy. Still, even with the advent of ultrahigh fields and of cryogenically cooled coils for in vivo research, the superposition of metabolic resonances arising from the brain remains a challenge. The present study explores the potential to tackle this milieu using a combination of two-dimensional (2D) NMR techniques, implemented on murine brains in vivo at 15.2 T and ex vivo at 14.1 T. While both experiments were affected by substantial inhomogeneous broadenings conveying distinct elongated lineshapes to the cross-peaks, the ability of increased fields to resolve off-diagonal resonances was clear. A comparison between the corresponding conventional and double quantum-filtered correlated spectroscopy traces enabled an improved assignment of in vivo resonances on the basis of more sensitive ex vivo 2D acquisitions, foremost on the basis of homonuclear cross-relaxation-driven correlations for peaks resonating downfield from water, and of heteronuclear correlations at natural abundance for the upfield protons. With the aid of such 2D correlations approximately 29 metabolites could be resolved and identified. This enhanced resolution was used to explore features related to the metabolites' diffusivities, their exposure to water, and their facility to undergo magnetization transfers to amide/amine/hydroxyl resonances. Cross-peaks from main murine brain biomolecules, including choline, creatine, γ-aminobutyric acid, N-acetyl aspartate, glutamine, and glutamate, showed enhancements in several of these various features, opening interesting vistas about metabolite compartmentalization as viewed by these 2D NMR experiments.

Deuterium imaging of the Warburg effect at sub-millimolar concentrations by joint processing of the kinetic and spectral dimensions.

Deuterium metabolic imaging (DMI) is a promising molecular MRI approach, which follows the administration of deuterated substrates and their metabolization. [6,6'-2 H2 ]-glucose for instance is preferentially converted in tumors to [3,3'-2 H2 ]-lactate as a result of the Warburg effect, providing a distinct resonance whose mapping using time-resolved spectroscopic imaging can diagnose cancer. The MR detection of low-concentration metabolites such as lactate, however, is challenging. It has been recently shown that multi-echo balanced steady-state free precession (ME-bSSFP) increases the signal-to-noise ratio (SNR) of these experiments approximately threefold over regular chemical shift imaging; the present study examines how DMI's sensitivity can be increased further by advanced processing methods. Some of these, such as compressed sensing multiplicative denoising and block-matching/3D filtering, can be applied to any spectroscopic/imaging methods. Sensitivity-enhancing approaches were also specifically tailored to ME-bSSFP DMI, by relying on priors related to the resonances' positions and to features of the metabolic kinetics. Two new methods are thus proposed that use these constraints for enhancing the sensitivity of both the spectral images and the metabolic kinetics. The ability of these methods to improve DMI is evidenced in pancreatic cancer studies carried at 15.2 T, where suitable implementations of the proposals imparted eightfold or more SNR improvement over the original ME-bSSFP data, at no informational cost. Comparisons with other propositions in the literature are briefly discussed.

Harnessing Water to Enhance Quadrupolar NMR Spectroscopy and Imaging.

17 O and 14 N are attractive targets for in vivo NMR spectroscopy and imaging, but low gyromagnetic ratios γ and fast spin relaxation complicate observations. This work explores indirect ways of detecting some of these sites with the help of proton-detected double resonance techniques. As standard coherence transfer methods are of limited use for such indirect detection, alternative routes for probing the quadrupolar spectra on 1 H were tested. These centered on modulating the broadening effects imparted onto protons adjacent to the low-γ species through J couplings through either continuous wave or spin-echo double-resonance decoupling/recoupling sequences. As in all cases, the changes imparted by these double-resonance strategies were small due to the fast relaxation undergone by the quadrupoles, the sensitivity of these approaches was amplified by transferring their effects onto the abundant water 1 H signal. These amplifications were mediated by the spontaneous exchanges that the labile 1 Hs bound to 17 O or 14 N undergo with the water protons. In experiments designed on the basis of double-resonance spin echoes, these enhancements were imparted by looping the transverse encodings together with multiple longitudinal storage periods, leading to decoupling-recoupling with exchange (D-REX) sequences. In experiments designed on the basis of continuous on/off quadrupolar decoupling, these solvent exchanges were incorporated into chemical-exchange saturation transfer schemes, leading to decoupling-recoupling with saturation transfer (D-REST) sequences. Both of these variants harnessed sizable proportions of the easily detectable water signals, in order to characterize the NMR spectra and/or to image with atomic-site specificity the 17 O and 14 N species.

Identification of variable stages in murine pancreatic tumors by a multiparametric approach employing hyperpolarized 13 C MRSI, 1 H diffusivity and 1 H T1 MRI.

This study explored the usefulness of multiple quantitative MRI approaches to detect pancreatic ductal adenocarcinomas in two murine models, PAN-02 and KPC. Methods assayed included 1 H T1 and T2 measurements, quantitative diffusivity mapping, magnetization transfer (MT) 1 H MRI throughout the abdomen and hyperpolarized 13 C spectroscopic imaging. The progress of the disease was followed as a function of its development; studies were also conducted for wildtype control mice and for mice with induced mild acute pancreatitis. Customized methods developed for scanning the motion- and artifact-prone mice abdomens allowed us to obtain quality 1 H images for these targeted regions. Contrasts between tumors and surrounding tissues, however, were significantly different. Anatomical images, T2 maps and MT did not yield significant contrast unless tumors were large. By contrast, tumors showed statistically lower diffusivities than their surroundings (≈8.3 ± 0.4 x 10-4 for PAN-02 and ≈10.2 ± 0.6 x 10-4 for KPC vs 13 ± 1 x 10-3 mm2 s-1 for surroundings), longer T1 relaxation times (≈1.44 ± 0.05 for PAN-02 and ≈1.45 ± 0.05 for KPC vs 0.95 ± 0.10 seconds for surroundings) and significantly higher lactate/pyruvate ratios by hyperpolarized 13 C MR (0.53 ± 0.2 for PAN-02 and 0.78 ± 0.2 for KPC vs 0.11 ± 0.04 for control and 0.31 ± 0.04 for pancreatitis-bearing mice). Although the latter could also distinguish early-stage tumors from healthy animal controls, their response was similar to that in our pancreatitis model. Still, this ambiguity could be lifted using the 1 H-based reporters. If confirmed for other kinds of pancreatic tumors this means that these approaches, combined, can provide a route to an early detection of pancreatic cancer.

Dynamic T2 mapping by multi-spin-echo spatiotemporal encoding.

PURPOSE: To develop a pulse sequence for acquiring robust, quantitative T2 relaxation maps in real time. METHODS: The pulse scheme relies on fully refocused spatiotemporally encoded multi-spin-echo trains, which provide images that are significantly less distorted than spin-echo echo planar imaging-based counterparts. This enables single-shot T2 mapping in inhomogeneity-prone regions. Another advantage of these schemes stems from their ability to interleave multiple scans in a reference-free manner, providing an option to increase sensitivity and spatial resolution with minimal motional artifacts. RESULTS: The method was implemented in preclinical and clinical scanners, where single-shot acquisitions delivered reliable T2 maps in ≤200 ms with ≈250 µm and ≈3 mm resolutions, respectively. Ca. 4 times higher spatial resolutions were achieved for the motion-compensated interleaved versions of these acquisitions, delivering T2 maps in ca. 10 s per slice. These maps were nearly indistinguishable from multi-scan relaxometric maps requiring orders-of-magnitude longer acquisitions; this was confirmed by mice head and real-time mice abdomen 7T scans performed following contrast-agent injections, as well as by 3T human brain and breast scans. CONCLUSION: This study introduced and demonstrated a new approach for acquiring rapid and quantitative T2 data, which is particularly reliable when operating at high fields and/or targeting heterogeneous organs or regions.

Controlled trial of an mHealth intervention to promote healthy behaviours in adolescence (TeenPower): Effectiveness analysis.

AIM: To evaluate the effectiveness on lifestyle change of an mHealth intervention to promote healthy behaviours in adolescence (TeenPower) and to analyse the predictors of the mHealth intervention effectiveness. DESIGN: This study is designed as a non-randomized controlled trial with a two-arm structure. METHODS: Adolescents of 12-16-year old were recruited from three school districts, with access to the Internet and smartphone/tablet devices. The intervention group was invited to engage in the mHealth intervention (TeenPower) for 6 months in addition to a school-based intervention. The control group only followed the school-based intervention. A repeated measures factorial ANOVA was used and the main effectiveness outcome was the lifestyle change measured by the adolescent lifestyle profile. RESULTS: The outcomes of the mHealth intervention (TeenPower) show a significant effect on nutrition (ƞ2 p  = 0.03, p = .03), positive life perspective (ƞ2 p  = 0.04, p = .01), and global lifestyle (ƞ2 p  = 0.02, p = .05), with a dropout rate of 62.1%. The analysis of the effectiveness predictors of the mHealth intervention suggested that older adolescents tended to show a significant increase in the rates of stress management (r = .40; p < .05). CONCLUSIONS: Although the considerable dropout rate, the mHealth intervention presented significant impact on multiple lifestyle domains, providing support for the effectiveness of mHealth interventions for health promotion as an add-on to standard interdisciplinary interventions. IMPACT: Adolescents must have the necessary and appropriate knowledge for the correct and responsible decision-making regarding their health and lifestyle. Innovative strategies (mHealth intervention) were used to promote healthy behaviours. This study evaluates the effectiveness of an mHealth intervention (TeenPower) specifically designed for adolescents. We found a significant impact in several lifestyle domains such as health responsibility, nutrition, positive life perspective, and global lifestyle.

Sensitivity-enhanced detection of non-labile proton and carbon NMR spectra on water resonances.

Chemical exchange saturation transfer (CEST) experiments enhance the NMR signals of labile protons by continuously transferring these protons' saturation to an abundant solvent pool like water. The present study expands these principles by fusing into these experiments homonuclear isotropic mixing sequences, enabling the water-enhanced detection of non-exchangeable species. Further opportunities are opened by the addition of coupling-mediated heteronuclear polarization transfers, which then impose on the water resonance a saturation stemming from non-labile heteronuclear species like 13C. To multiplex the ensuing experiments, these relayed approaches are combined with time-domain schemes involving multiple Ramsey-labeling experiments imparting the frequencies of the non-labile sites on the water resonance, via chemical exchange. 13C and 1H NMR spectra were detected in this fashion with about two-fold SNR amplification vis-à-vis conventionally detected spectroscopies. When combined with non-uniform sampling principles, this methodology thus becomes a sensitive alternative to detect non-exchangeable species in biomolecules. Still, multiple parameters including the scalar couplings and solvent exchange rates, will affect the efficiency and consequently the practicality of the overall experiment.

Heteronuclear 1D and 2D NMR Resonances Detected by Chemical Exchange Saturation Transfer to Water.

A method to detect NMR spectra from heteronuclei through the modulation that they impose on a water resonance is exemplified. The approach exploits chemical exchange saturation transfers, which can magnify the signal of labile protons through their influence on a water peak. To impose a heteronuclear modulation on water, an HMQC-type sequence was combined with the FLEX approach. 1D 15 N NMR spectra of exchanging sites could thus be detected, with about tenfold amplifications over the 15 N modulations afforded by conventionally detected HMQC NMR spectroscopy. Extensions of this approach enable 2D heteronuclear acquisitions on directly bonded 1 H-15 N spin pairs, also with significant signal amplification. Despite the interesting limits of detection that these signal enhancements could open in NMR spectroscopy, these gains are constrained by the rates of solvent exchange of the targeted heteronuclear pairs, as well as by spectrometer instabilities affecting the intense water resonances detected in these experiments.

Distribution of polyelectrolytes and counterions upon polyelectrolyte complexation.

HYPOTHESIS: Understanding polyelectrolyte complexation remains limited due to the absence of a systematic methodology for analyzing the distribution of components between the polyelectrolyte complex (PEC) and the dilute phases. EXPERIMENTS: We developed a methodology based on NMR to quantify all components of solid-like PECs and their supernatant phases formed by mixing different ratios of poly(allylamine hydrochloride) (PAH) and poly(acrylic acid)-sodium salt (PAA). This approach allowed for determining relative and absolute concentrations of polyelectrolytes in both phases by 1H NMR studies. Using 23Na and 35Cl NMR spectroscopy we measured the concentration of counterions in both phases. FINDINGS: Regardless of the mixing ratio of the polyelectrolytes the PEC is charge-stoichiometric, and any excess polyelectrolytes to achieve charge stoichiometry remains in the supernatant phase. The majority of counterions were found in the supernatant phase, confirming counterion release being a major thermodynamic driving force for PEC formation. The counterion concentrations in the PEC phase were approximately twice as high as in the supernatant phase. The complete mass balance of PEC formation could be determined and translated into a molecular picture. It appears that PAH is fully charged, while PAA is more protonated, so less charged, and some 10% extrinsic PAH-Cl- pairs are present in the complex.

Stationary wavelet denoising of solid-state NMR spectra using multiple similar measurements.

Accumulating several scans of free induction decays is always needed to improve the signal-to-noise ratio of NMR spectra, especially for the low gyromagnetic ratio solid-state NMR. In this study, we present a new denoising approach based on the correlations between multiple similar NMR spectra. Contrary to the simple averaging of multiple scans or denoising the final averaged spectrum, we propose a Wavelet-based Denoising technique for Multiple Similar scans(WDMS). Firstly, the stationary wavelet transform is applied to decompose every spectrum into approximation coefficients and detail coefficients. Then, the detail coefficients are multiplied by weights calculated based on Pearson's correlation coefficient and structural similarity index between approximation coefficients of different spectra. Finally, the average of these detailed components is used to denoise the spectra. The proposed method is carried on the assumption that noise between multiple spectra is uncorrelated while peak signal information is similar between different spectra, thus preserving the possibility of applying further processing to the data. As a demonstration, the standard wavelet denoise is applied to the WDMS-processed spectra, achieving a further increase in the S/N ratio. We confirm the reliability of the denoising approach based on multiple scans on 1D/2D solid-state MAS/static NMR spectra. In addition, we also show that this method can be used to deal with a single Car-Purcell-Meiboom-Gill (CPMG) echo train.

Engineering of Adhesion at Metal-Poly(lactic acid) Interfaces by Poly(dopamine): The Effect of the Annealing Temperature.

Control over adhesion at interfaces from strong bonding to release between thermoplastic polymers (TPs) and metal oxides is highly significant for polymer composites. In this work, we showcase a simple and inexpensive method to tune adhesion between a TP of growing interest, poly(lactic acid) (PLA), and two commercial metal alloys, based on titanium and stainless steel. This is realized by coating titanium and stainless steel wires with polydopamine (PDA), thermally treating them under vacuum at temperatures ranging from 25 to 250 °C, and then comolding them with PLA to form pullout specimens for adhesion tests. Pullout results indicate that PDA coatings treated at low temperatures up to a given threshold significantly improve adhesion between PLA and the metals. Conversely, at higher PDA annealing temperatures beyond the threshold, interfacial bonding gradually declines. The excellent control over interfacial adhesion is attributed to the thermally induced transformation of PDA. In this work, we show using thermogravimetric analysis, X-ray photoelectron spectroscopy, Fourier transform infrared, and 13C solid-state NMR that the extent of the thermal transformation is dependent on the annealing temperature. By selecting the annealing temperature, we vary the concentration of primary amine and hydroxyl groups in PDA, which influences adhesion at the metal/PLA interface. We believe that these findings contribute to optimizing and broadening the applications of PDA in composite materials.

Building Bridges for "Palliative Care-in-Place": Development of a mHealth Intervention for Informal Home Care.

Background: In Palliative Care (PC), family and close people are an essential part of provision of care. They assume highly complex tasks for which they are not prepared, with considerable physical, psychological, social and economic impact. Informal Caregivers (ICs) often falter in the final stage of life and develop distress, enhancing emotional burden and complicated grief. The lack of available and accessible in-person counselling resources is often reported by ICs. Online resources can promote early access to help and support for patient-IC dyads in palliative care. The primary aim of this research is to co-design, develop and test the feasibility of the Help2Care-PAL mHealth app that addresses the needs of ICs of palliative patients cared for at home. This Digital Health Intervention (DHI) in palliative care will be used for education, symptom management, communication and decision-making, to enhance Quality of Life (QoL) of patients and ICs, fostering anticipatory grief and the reach and efficiency of services. Methods: This study will use an iterative co-design process and convergent mixed-methods design, following the MORECare consensus for developing a complex intervention. Construction of the DHI will follow four main phases: (I) a needs assessment (a cross-sectional survey, individual interviews with ICs and focus groups with professionals with community palliative care experience); (II) design and co-production of mHealth materials and interventions to support ICs; (III) the development of a mHealth app; and (IV) usability and feasibility of the mHealth app. The Help2Care-PAL platform seeks to build resources from the perspectives and needs of both family dyads and nursing professionals working in the field of community palliative care. User-centeredness will be ensured by the active participation of patient-IC dyads and professionals of the palliative care community. Discussion: This mixed-method study will offer new insights on needs and expectations of patient-IC dyads and nurses in community palliative care regarding caregiving preparedness and online health resources. Through the implementation of an adaptive digital tool, we aim to improve access to palliative care family support, which is highly linked with the wellbeing of patients and especially new ICs.

A denoising method for multidimensional magnetic resonance spectroscopy and imaging based on compressed sensing.

Both in spectroscopy and imaging, t1-noise arising from instabilities such as temperature alterations, field-related frequency drifts, electronic and sample-spinning instabilities, or motions in in vivo experiments, affects many 2D Magnetic Resonance experiments. This work introduces a post-processing method that aims to attenuate t1-noise, by suitably averaging multiple signals/representations that have been reconstructed from the sampled data. The ensuing Compressed Sensing Multiplicative (CoSeM) denoising starts from a fully sampled 2D MR data set, discards random indirect-domain points, and makes up for these missing, masked data, by a compressed sensing reconstruction of the now incompletely sampled 2D data set. This procedure is repeated for multiple renditions of the masked data -some of which will have been more strongly affected by t1-noise than others. This leads to a large set of 2D NMR spectra/images compatible with the collected data; CoSeM chooses out of these those renditions that reduce the noise according to a suitable criterion, and then sums up their spectra/images leading to a reduction in t1-noise. The performance of the method was assessed in synthetic data, as well as in numerous different experiments: 2D solid and solution state NMR, 2D localized MRS of live brains, and 2D abdominal MRI. Throughout all these data, CoSeM processing evidenced 2-3 fold increases in SNR, without introducing biases, false peaks, or spectral/image blurring. CoSeM also retains a quantitative linearity in the information -allowing, for instance, reliable T1 inversion-recovery MRI mapping experiments.

A Clinical Decision Support System for Remote Monitoring of Cardiovascular Disease Patients: A Clinical Study Protocol.

Introduction: Cardiovascular diseases (CVD) are the leading cause of death globally, taking an estimated 17. 9 million lives each year. Cardiac rehabilitation is shown to reduce mortality and hospital readmissions, while improving physical fitness and quality of life. Despite the recommendations and proven benefits, acceptance and adherence remain low. Mobile health (mHealth) solutions may contribute to more personalized and tailored patient recommendations according to their specific needs. This study protocol aims to assess the effectiveness of a user-friendly, comprehensive Clinical Decision Support System (CDSS) for remote patient monitoring of CVD patients, primarily on the reduction of recurrent cardiovascular events. Methods and Analysis: The study will follow a multicenter randomized controlled design involving two cardiology units in the Center Region of Portugal. Prospective CVD patients will be approached by the healthcare staff at each unit and checked for eligibility according to the predefined inclusion/exclusion criteria. The CDSS will suggest a monitoring plan for the patient, will advise the mHealth tools (apps and wearables) adapted to patient needs, and will collect data. The clinical study will start in January 2023. Discussion: The success of the mHeart.4U intervention will be a step toward the use of technological interfaces as an integrating part of CR programs. Ethics and Dissemination: The study will undergo ethical revision by the Ethics Board of the two hospital units where the study will unfold. The study was registered in ClinicalTrials.gov on 18th January 2022 with the number NCT05196802. The study findings will be published in international peer-reviewed scientific journals and encounters and in a user-friendly manner to the society.

Development of an mHealth Platform for Adolescent Obesity Prevention: User-Centered Design Approach.

Obesity is a chronic condition that influences the quality of life of patients and families while increasing the economic burden for the world population. Multidisciplinary prevention programs are crucial to address it, allowing an early introduction of healthy behaviors into daily habits. Mobile health interventions provide adequate support for these programs, especially considering the gamification techniques used to promote users' engagement. TeenPower is a multidisciplinary mHealth intervention program conducted in Portugal during 2018 to empower adolescents, promoting healthy behaviors while preventing obesity. An agile software development process was applied to the development of the digital platform that holds a web-based application and a mobile application. We also propose a model for future developments based on the user-centered design approach adopted for this development and the assessment conducted in each phase. The user-centered design approach model proposed has three distinct phases: (1) design study; (2) pre-production usability tests; and (3) post-production data. Phase 1 allowed us to obtain the high-fidelity version of the graphical user interfaces (n = 5). Phase 2 showed a task completion success rate of 100% (n = 5). Phase 3 was derived from statistical analysis of the usage of the platform by real end users (n = 90). We achieved an average retention rate of 35% (31 out of 90 participants). Each technique has provided input for the continuous design and improvement of the platform. This allowed the creation of a tailored platform that could meet users' expectations. Nevertheless, the retention rate decreased significantly over a short period of time, revealing the need for further work in the improvement of the gamification experience.

Improving MRI's slice selectivity in the presence of strong, metal-derived inhomogeneities.

PURPOSE: To develop schemes that deliver faithful 2D slices near field heterogeneities of the kind arising from non-ferromagnetic metal implants, with reduced artifacts and shorter scan times. METHODS: An excitation scheme relying on cross-term spatio-temporal encoding (xSPEN) was used as basis for developing the new inhomogeneity-insensitive, slice-selective pulse scheme. The resulting Fully refOCUSED cross-term SPatiotemporal ENcoding (FOCUSED-xSPEN) approach involved four adiabatic sweeps. The method was evaluated in silico, in vitro and in vivo using mice models, and compared against a number of existing and of novel alternatives based on both conventional and swept RF pulses, including an analogous method based on LASER's selectivity spatial selectivity. RESULTS: Calculations and experiments confirmed that multi-sweep derivatives of xSPEN and LASER can deliver localized excitation profiles, centered at the intended positions and endowed with enhanced immunity to B0 and B1 distortions. This, however, is achieved at the expense of higher SAR than non-swept counterparts. Furthermore, single-shot FOCUSED-xSPEN and LASER profiles covered limited off-resonance ranges. This could be extended to bands covering arbitrary off-resonance values with uniform slice widths, by looping the experiments over a number of scans possessing suitable transmission and reception offsets. CONCLUSIONS: A series of novel approaches were introduced to select slices near metals, delivering robustness against Bo and B1+ field inhomogeneities.

Training load and well-being status variations of elite futsal players across a full season: Comparisons between normal and congested weeks.

The aim of this study was two-fold: (i) to analyze the variations of internal load (IL) and well-being between normal and congested weeks in professional futsal (also known as indoor soccer) players and (ii) to analyze variations in training days (MD-1, MD-2 and MD-3 [matchday -1, -2, and -3, respectively]) within weeks. Twenty male professional futsal players (age: 27.8 ±â€¯5.7 years old; height: 173.8 ±â€¯5.6 cm; weight: 71.5 ±â€¯7.9 kg) that participated in the first futsal league of Portugal were monitored in all training sessions and matches during a full season. Session ratings of perceived exertion and a Hooper questionnaire (to measure muscle soreness, stress, fatigue, and sleep) were used to monitor the IL and well-being, respectively. Almost certain large decreases in IL during congested weeks in training session MD-3 were observed (-27.7%, [-35.8;-18.6]). Likely moderate decreases in IL during congested weeks in the MD-2 session were also verified (-9.9%, [-17.2;-2.0]). Regarding muscle soreness, almost certain large decreases were found during congested weeks on MD-3 (-21.2%, [-30.0;-11.3]) and MD-1 (-17.3%, [-26.6;-6.9]). Similar evidence was found for fatigue level, as almost certain large decreases were observed during congested weeks on MD-3 (-23.5%, [-32.9;-12.7]) and MD-1 (-16.6%, [-24.8;-7.4]). Within-week changes during normal weeks revealed almost certain large increases in internal load on MD-2 (93.4%, [78.6;109.3]) and MD-3 (106.7%, [88.2;127.0]) in comparison with MD-1. Also, during congested weeks, almost certain large increases of internal load were observed on MD-2 (77.2%, [53.4;104.7]) and MD-3 (65.7%, [42.8;92.3]) in comparison with MD-1. The major findings of this study revealed meaningful increases of IL, muscle soreness, and fatigue in normal weeks in comparison to congested ones. Within-week changes were also observed, with the greatest loads being observed on MD-3 during normal weeks and on MD-2 during congested weeks.