NlpC/P60 peptidoglycan hydrolases of Trichomonas vaginalis have complementary activities that empower the protozoan to control host-protective lactobacilli.

Trichomonas vaginalis is a human protozoan parasite that causes trichomoniasis, a prevalent sexually transmitted infection. Trichomoniasis is accompanied by a shift to a dysbiotic vaginal microbiome that is depleted of lactobacilli. Studies on co-cultures have shown that vaginal bacteria in eubiosis (e.g. Lactobacillus gasseri) have antagonistic effects on T. vaginalis pathogenesis, suggesting that the parasite might benefit from shaping the microbiome to dysbiosis (e.g. Gardnerella vaginalis among other anaerobes). We have recently shown that T. vaginalis has acquired NlpC/P60 genes from bacteria, expanding them to a repertoire of nine TvNlpC genes in two distinct clans, and that TvNlpCs of clan A are active against bacterial peptidoglycan. Here, we expand this characterization to TvNlpCs of clan B. In this study, we show that the clan organisation of NlpC/P60 genes is a feature of other species of Trichomonas, and that Histomonas meleagridis has sequences related to one clan. We characterized the 3D structure of TvNlpC_B3 alone and with the inhibitor E64 bound, probing the active site of these enzymes for the first time. Lastly, we demonstrated that TvNlpC_B3 and TvNlpC_B5 have complementary activities with the previously described TvNlpCs of clan A and that exogenous expression of these enzymes empower this mucosal parasite to take over populations of vaginal lactobacilli in mixed cultures. TvNlpC_B3 helps control populations of L. gasseri, but not of G. vaginalis, which action is partially inhibited by E64. This study is one of the first to show how enzymes produced by a mucosal protozoan parasite may contribute to a shift on the status of a microbiome, helping explain the link between trichomoniasis and vaginal dysbiosis. Further understanding of this process might have significant implications for treatments in the future.

Lactobacillus gasseri and Gardnerella vaginalis produce extracellular vesicles that contribute to the function of the vaginal microbiome and modulate host-Trichomonas vaginalis interactions.

Trichomonas vaginalis is an extracellular protozoan parasite of the human urogenital tract, responsible for a prevalent sexually transmitted infection. Trichomoniasis is accompanied by a dysbiotic microbiome that is characterised by the depletion of host-protective commensals such as Lactobacillus gasseri, and the flourishing of a bacterial consortium that is comparable to the one seen for bacterial vaginosis, including the founder species Gardnerella vaginalis. These two vaginal bacteria are known to have opposite effects on T. vaginalis pathogenicity. Studies on extracellular vesicles (EVs) have been focused on the direction of a microbial producer (commensal or pathogen) to a host recipient, and largely in the context of the gut microbiome. Here, taking advantage of the simplicity of the human cervicovaginal microbiome, we determined the molecular cargo of EVs produced by L. gasseri and G. vaginalis and examined how these vesicles modulate the interaction of T. vaginalis and host cells. We show that these EVs carry a specific cargo of proteins, which functions can be attributed to the opposite roles that these bacteria play in the vaginal biome. Furthermore, these bacterial EVs are delivered to host and protozoan cells, modulating host-pathogen interactions in a way that mimics the opposite effects that these bacteria have on T. vaginalis pathogenicity. This is the first study to describe side-by-side the protein composition of EVs produced by two bacteria belonging to the opposite spectrum of a microbiome and to demonstrate that these vesicles modulate the pathogenicity of a protozoan parasite. Such as in trichomoniasis, infections and dysbiosis co-occur frequently resulting in significant co-morbidities. Therefore, studies like this provide the knowledge for the development of antimicrobial therapies that aim to clear the infection while restoring a healthy microbiome.

Challenges and opportunities for access to Advanced Therapy Medicinal Products in Brazil.

BACKGROUND AIMS: The marketing authorization of Advanced Therapy Medicinal Products (ATMPs) in Brazil is recent. The features of these therapies impose specialized regulatory action and are consequently challenging for developers. The goal of this study was to identify the industry's experience in clinical development, marketing authorization and access to ATMPs through the Unified Health System (SUS, acronym in Portuguese), from a regulatory perspective. METHODS: A survey containing structured questions was conducted among research participants who work at companies that commercialize ATMPs. A descriptive analysis was performed. RESULTS: We invited 15 foreign pharmaceutical companies, of which 10 agreed to participate. Overall, participants assessed that Brazil has a well-established regulatory system, especially the sanitary registration by the National Health Surveillance Agency (Anvisa), which ensures the quality, safety, and efficacy of the products. The Agency's good interaction with the regulated sector, the harmonization of sanitary and ethical assessment systems with other countries, and the analysis time in the biosafety assessment of Genetically Modified Organisms (GMOs) stand out as positive in industry's evaluation. On the other hand, it is important to advance the pricing regulation for these products since Brazilian regulations do not establish specific criteria for ATMP. One of the biggest challenges is the difficulty for the SUS in reimbursing these very high-cost therapies, especially using current Health Technology Assessment (HTA) methods. CONCLUSIONS: Considering the increasing number of approvals of cell and gene therapies in Brazil in the coming years, a close dialogue between the industry and the public sector is recommended to advance regulatory improvements (pricing and HTA). Additionally, the construction of policies to promote the national Health Economic-Industrial Complex, based on a mission-oriented vision that encourages innovative models of financing, especially those that consider risk-sharing and co-financing technologies, will help provide the population with universal, equitable and sustainable access to ATMP in the SUS.

Development and Characterization of Monoolein-Based Liposomes of Carvacrol, Cinnamaldehyde, Citral, or Thymol with Anti-Candida Activities.

There is an increasing need for novel drugs and new strategies for the therapy of invasive candidiasis. This study aimed to develop and characterize liposome-based nanoparticles of carvacrol, cinnamaldehyde, citral, and thymol with anti-Candida activities. Dioctadecyldimethylammonium bromide- and monoolein-based liposomes in a 1:2 molar ratio were prepared using a lipid-film hydration method. Liposomes were assembled with equal volumes of liposomal stock dispersion and stock solutions of carvacrol, cinnamaldehyde, citral, or thymol in dimethyl sulfoxide. Cytotoxicity was tested on RAW 264.7 macrophages. In vitro antifungal activity of liposomes with phytocompounds was evaluated according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology using clinical isolates of Candida albicans, Candida auris, Candida dubliniensis, and Candida tropicalis Finally, the ability of macrophage cells to kill Candida isolates after addition of phytocompounds and their nanoparticles was determined. Nanoparticles with 64 µg/ml of cinnamaldehyde, 256 µg/ml of citral, and 128 µg/ml of thymol had the best characteristics among the formulations tested. The highest encapsulation efficiencies were achieved with citral (78% to 83%) and carvacrol (66% to 71%) liposomes. Carvacrol and thymol in liposome-based nanoparticles were nontoxic regardless of the concentration. Moreover, carvacrol and thymol maintained their antifungal activity after encapsulation, and there was a significant reduction (∼41%) of yeast survival when macrophages were incubated with carvacrol or thymol liposomes. In conclusion, carvacrol and thymol liposomes possess high stability, low cytotoxicity, and antifungal activity that act synergistically with macrophages.

How do swimmers pace the 400 m freestyle and what affects the pacing pattern?

To describe the pacing pattern of elite 400 m freestyle swimmers, understand how athletes select the first half compared to their 200-m race pace, and correlate critical speed (CS) and D' with pacing parameters. The best all-time 400 m performances were obtained, and 200 m performances were retrieved from the same competition. Twenty-four performances were selected and, when possible, 50 m-splits were obtained. CS and D' were calculated based on the 200 m and 400 m performances. Most athletes adopted a parabolic pacing pattern. Performance in the 400 m correlated with that of the 200 m (rho = 0.418, p = 0.042), the % difference between 200 m performance and 200 m split correlated with CS (rho = -0.595, p = 0.002) and with D' (rho = 0.808, p < 0.001). The correlation between D' and % difference between average speed and CS was nearly perfect (rho = 1.000, p < 0.001). Recommendations of having the first half of the 400 m 4-5 s slower than their 200 m race do not seem to be appropriate for all swimmers and depends on physiological abilities.

Vaginal dysbiotic bacteria act as pathobionts of the protozoal pathogen Trichomonas vaginalis.

Trichomoniasis, a prevalent sexually transmitted infection caused by the protozoan parasite Trichomonas vaginalis, is often accompanied by a vaginal dysbiotic microbiota of pathogenic potential. Our objective was to investigate whether these dysbiotic bacteria act as pathobionts of T. vaginalis infection by altering pathogenic capabilities of the parasite, particularly in regard to adhesion to vaginal substrates and viability of human ectocervical cells. Assays interrogated the performance of T. vaginalis adhesion to biofilm produced by vaginal dysbiotic bacteria and whether these bacteria were capable of altering the ability of the parasite to bind to mucins and cells. The binding activities of T. vaginalis were quantified by flow cytometry. Host cell viability and apoptosis, as affected by T. vaginalis with or without the bacteria, were also measured experimentally. An in vitro biofilm was shown to provide adhesion for T. vaginalis. The binding of parasites to mucins and cells was modulated by the vaginal dysbiotic bacteria. Parasite cytoadhesion was significantly increased by these bacteria. In addition, these bacteria enhanced the pathogenic effects of the parasite to host cells. Together, this study showed that dysbiotic bacteria accompanying T. vaginalis infection in the vagina function as pathobionts as they are capable of enhancing the pathogenic capabilities of this parasite. This study highlights the importance of understanding the contribution of the vaginal microbiome to trichomoniasis.

Does a 4-week training period with hand paddles affect front-crawl swimming performance?

This study investigated the effects of a 4-week training with hand paddles (HPD) on front-crawl swimming performance (SP), clean swimming speed (SPEED), stroke rate (SR), stroke length (SL) and tethered force (TF). Twenty swimmers (10 men and 10 women) were paired according to performance and gender, and were randomly assigned to control (CON, 22.4 ± 2.3 years) or HPD (21.8 ± 1.9 years) groups. During 4 weeks both groups performed the same training, except for a sprint training set (3 times/week, 10 × 10 strokes all-out, 1-min rest) completed with (HPD = 320 cm2) and without (CON) paddles. Afterwards, both groups performed the same training over a 2-week taper period. SP, SPEED, SR, SL and TF were assessed before (PRE) and after the 4-week period (POST), after the first (T1) and second taper weeks (T2). Swimmers rated their perceived exertion for the sprint training set (RPETS) and the training session for determining internal training load (ITL). SP, SPEED, SR, SL and TF did not change from PRE to POST, T1 and T2. ITL and RPETS were not different between groups. Training 4 weeks with HPD does not affect swimming performance, so the use of HPD remains unsupported in such period.

Cooperative Interactions between Trichomonas vaginalis and Associated Bacteria Enhance Paracellular Permeability of the Cervicovaginal Epithelium by Dysregulating Tight Junctions.

The human protozoan Trichomonas vaginalis is the causative agent of trichomoniasis, a prevalent sexually transmitted infection, which is accompanied by a species-diversified vaginal microbiota named community state type IV (CST-IV). Coincidently, CST-IV includes species associated with bacterial vaginosis (e.g. Gardnerella vaginalis, Atopobium vaginae, and Prevotella bivia). Both diseases are linked to the transmission of human immunodeficiency virus (HIV) and preterm birth, which complications are likely to result from the disruption of the cervicovaginal epithelial barrier. Here, we show that paracellular permeability of fluorescein isothiocyanate (FITC)-dextran through a monolayer of human ectocervical cells (hECs) is increased as a consequence of the activity of T. vaginalis and the aforementioned species of CST-IV bacteria cooperatively. T. vaginalis enhances paracellular permeability of hECs two times more than the individual bacterial species, by up to ∼10% versus ∼5%, respectively. However, any two or all three bacterial species are capable of synergizing this effect. T. vaginalis and the bacteria together increase the paracellular permeability of hECs by ∼50%, which is 5 to 10 times more than the results seen with the protozoan or bacteria alone. This effect is accompanied by enhancement of phosphatase activity, while phosphatase inhibition results in preservation of the integrity of the ectocervical cell monolayer. In addition, these microorganisms induce changes in the expression of tight junction proteins, particularly occludin, and of proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α). Together, our findings establish that cooperative interactions between CST-IV bacteria and T. vaginalis enhance the paracellular permeability of the cervicovaginal epithelium by disturbing the integrity of the tight junction complex. Our study results highlight the importance of understanding the contribution of the vaginal microbiota to trichomoniasis.

A Cell Surface Aggregation-Promoting Factor from Lactobacillus gasseri Contributes to Inhibition of Trichomonas vaginalis Adhesion to Human Vaginal Ectocervical Cells.

Trichomoniasis, a prevalent sexually transmitted infection, is commonly symptomatic in women. The causative agent is Trichomonas vaginalis, an extracellular protozoan parasite. The host-protective mechanisms and molecules of vaginal lactobacilli that counteract this pathogen are largely unknown. This study examines the inhibition promoted by Lactobacillus gasseri against the adhesion of T. vaginalis to host cells, a critical virulence aspect of this pathogen. We observed that the vaginal strain L. gasseri ATCC 9857 is highly inhibitory by various contact-dependent mechanisms and that surface proteins are largely responsible for this inhibitory phenotype. We found that the aggregation-promoting factor APF-2 from these bacteria significantly contributes to inhibition of the adhesion of T. vaginalis to human vaginal ectocervical cells. Understanding the molecules and mechanisms used by lactobacilli to protect the host against T. vaginalis might help in the development of novel and specific therapeutic strategies that take advantage of the natural microbiota.

High variability within Candida albicans transcription factor RLM1: Isolates from vulvovaginal infections show a clear bias toward high molecular weight alleles.

Previous studies have correlated the severity of recurrent vulvovaginal Candida infections (VVC) and balanitis in patients from China with the presence of some dominant genotypes at the ORF RLM1. Here we tested VVC vs non-VVC isolates from Portugal, Brazil and Greece and, although the same genotypes were identified in VVC isolates, they were present in only five out of 150 strains. However, this analysis showed that VVC isolates presented a higher percentage of genotypes with similar high molecular weight alleles, in comparison with strains isolated from other biological sources.

Effect of hand paddles and parachute on butterfly coordination.

This study investigated the effects of hand paddles, parachute and hand paddles plus parachute on the inter-limb coordination of butterfly swimming. Thirteen male swimmers were evaluated in four random maximal intensity conditions: without equipment, with hand paddles, with parachute and with hand paddles + parachute. Arm and leg stroke phases were identified by 2D video analysis to calculate the total time gap (T1: time between hands' entry in the water and high break-even point of the first undulation; T2: time between the beginning of the hand's backward movement and low break-even point of the first undulation; T3: time between the hand's arrival in a vertical plane to the shoulders and high break-even point of the second undulation; T4: time between the hand's release from the water and low break-even point of the second undulation). The swimming velocity was reduced and T1, T2 and T3 increased in parachute and hand paddles + parachute. No changes were observed in T4. Total time gap decreased in parachute and hand paddles + parachute. It is concluded that hand paddles do not influence the arm-to-leg coordination in butterfly, while parachute and hand paddles + parachute do change it, providing a greater propulsive continuity.

Box H/ACA snoRNAs are preferred substrates for the trimethylguanosine synthase in the divergent unicellular eukaryote Trichomonas vaginalis.

The 2,2,7-trimethylguanosine caps of eukaryal snRNAs and snoRNA are formed by the enzyme Tgs1, which catalyzes sequential guanine-N2 methylations of m(7)G caps. Atypically, in the divergent unicellular eukaryote Trichomonas vaginalis, spliceosomal snRNAs lack a guanosine cap and the recombinant T. vaginalis trimethylguanosine synthase (TvTgs) produces only m(2,7)G in vitro. Here, we show by direct metabolic labeling that endogenous T. vaginalis RNAs contain m(7)G, m(2,7)G, and m(2,2,7)G caps. Immunodepletion of TvTgs from cell extracts and TvTgs add-back experiments demonstrate that TvTgs produces m(2,7)G and m(2,2,7)G caps. Expression of TvTgs in yeast tgs1Δ cells leads to the formation of m(2,7)G and m(2,2,7)G caps and complementation of the lethality of a tgs1Δ mud2Δ strain. Whereas TvTgs is present in the nucleus and cytosol of T. vaginalis cells, TMG-containing RNAs are localized primarily in the nucleolus. Molecular cloning of anti-TMG affinity-purified T. vaginalis RNAs identified 16 box H/ACA snoRNAs, which are implicated in guiding RNA pseudouridylation. The ensemble of new T. vaginalis H/ACA snoRNAs allowed us to predict and partially validate an extensive map of pseudouridines in T. vaginalis rRNA.

Unconventional features in the transcription and processing of spliceosomal small nuclear RNAs in the protozoan parasite Trichomonas vaginalis.

Trichomonas vaginalis is a medically important protozoan parasite, and a deep-branching, evolutionarily divergent unicellular eukaryote that has conserved several key features of eukaryotic gene expression. Trichomonas vaginalis possesses a metazoan/plant-like capping apparatus, mRNAs with a cap 1 structure and spliceosomes containing the five small nuclear RNAs (snRNAs). However, in contrast to metazoan and plant snRNAs, the structurally conserved T. vaginalis snRNAs were initially identified as lacking the canonical guanosine cap nucleotide. To explain this unusual condition, we sought to investigate transcriptional and processing features of the spliceosomal snRNAs in this protist. Here, we show that T. vaginalis spliceosomal snRNA genes mostly lack typical eukaryotic promoters. In contrast to other eukaryotes, the putative TATA box in the T. vaginalis U6 snRNA gene was found to be dispensable for transcription or RNA polymerase selectivity. Moreover, U6 transcription in T. vaginalis was virtually insensitive to tagetitoxin compared with other cellular transcripts produced by the same RNA polymerase III. Most important and unexpected, snRNA transcription in T. vaginalis appears to bypass capping as we show that these transcripts retain their original 5'-triphosphate groups. In conclusion, transcription and processing of spliceosomal snRNAs in T. vaginalis deviate considerably from the conventional rules of other eukaryotes.

Candida albicans chitinase 3 with potential as a vaccine antigen: production, purification, and characterisation.

Chitinases are widely studied enzymes that have already found widespread application. Their continued development and valorisation will be driven by the identification of new and improved variants and/or novel applications bringing benefits to industry and society. We previously identified a novel application for chitinases wherein the Candida albicans cell wall surface chitinase 3 (Cht3) was shown to have potential in vaccine applications as a subunit antigen against fungal infections. In the present study, this enzyme was investigated further, developing production and purification protocols, enriching our understanding of its properties, and advancing its application potential. Cht3 was heterologously expressed in Pichia pastoris and a 4-step purification protocol developed and optimised: this involves activated carbon treatment, hydrophobic interaction chromatography, ammonium sulphate precipitation, and gel filtration chromatography. The recombinant enzyme was shown to be mainly O-glycosylated and to retain the epitopes of the native protein. Functional studies showed it to be highly specific, displaying activity on chitin, chitosan, and chito-oligosaccharides larger than chitotriose only. Furthermore, it was shown to be a stable enzyme, exhibiting activity, and stability over broad pH and temperature ranges. This study represents an important step forward in our understanding of Cht3 and contributes to its development for application.

The adherence of Trichomonas vaginalis to host ectocervical cells is influenced by lactobacilli.

OBJECTIVES: Trichomoniasis is a common sexually transmitted disease, and adhesion of the pathogen Trichomonas vaginalis to the host vaginal cells is the first step in establishing infection. For this to happen, the pathogen has to overcome a natural protective barrier composed mostly of lactobacilli. The objective of this study was to understand the role of lactobacilli in the adhesion of T vaginalis to host cells. METHODS: Adhesion assays were carried out by incubating vaginal epithelial cells (VECs) with T vaginalis and lactobacilli together and compared with non-lactobacilli recipient controls. By varying incubation parameters and testing several microbial isolates, the number of pathogens that adhered to the VECs was determined by flow cytometry. RESULTS: Overall, but with few exceptions, lactobacilli caused inhibition of T vaginalis adhesion to a variable degree. Lactobacillus gasseri ATCC 9857 and CBI3 (ambiguous Lactobacillus plantarum or Lactobacillus pentosus) caused the highest level of parasite adhesion inhibition and enhancement, respectively. These isolates of Lactobacillus can profoundly alter the adhesive properties of low-adherent and high-adherent strains of T vaginalis in a dose-dependent manner. Additionally, the effects of lactobacilli on T vaginalis adhesion are strictly contact-dependent, and surface lipoglycans of T vaginalis are most likely not involved in this modulation of adhesion mediated by the bacteria. CONCLUSIONS: Lactobacilli can modulate adhesion of T vaginalis by significantly modifying the natural adhesive properties of various T vaginalis strains. This study highlights the importance of considering the role of the vaginal microbiota in the pathogenesis of trichomoniasis.

Acute responses of biomechanical parameters to different sizes of hand paddles in front-crawl stroke.

This study investigated the acute effects of different sizes of paddles on the force-time curve during tethered swimming and swimming velocity in front-crawl stroke. Fourteen male swimmers (20.0 ± 3.7 years; 100-m best time: 53.70 ± 0.87 s) performed two 10-s maximal efforts in tethered swimming to obtain peak force, average force, impulse, rate of force development, stroke duration and time to peak force. Swimming velocity, stroke rate and stroke length were obtained from two 25-m maximal swims. Both tests were repeated in five conditions: free swimming, wearing small (280 cm (2) ), medium (352 cm (2) ), large (462 cm (2) ) and extra-large (552 cm (2) ) hand paddles. Compared to free swimming, paddles provided significant increases of peak force (medium: 11.5%, large: 16.7%, extra-large: 21.7%), impulse (medium: 15.2%, large: 22.4%, extra-large: 30.9%), average force (medium: 5.1%, large: 7.5%), rate of force development (extra-large: 11.3%), stroke duration (medium: 9.3%, large: 11.8%, extra-large: 18.5%), time to peak force (medium: 11.1%, large: 15.9%, extra-large: 22.1%), swimming velocity (medium: 2.2%, large: 3.2%, extra-large: 3.7%) and stroke length (medium: 9.0%, large: 9.0%, extra-large: 14.8%), while stroke rate decreased (medium: -6.2%, large: -5.5%, extra-large: -9.5%). It is concluded that medium, large and extra-large paddles influence the force-time curve and change swimming velocity, suggesting these sizes may be useful for force development in water.

Effects of Zn-ZnO Core-Shell Nanoparticles on Antimicrobial Mechanisms and Immune Cell Activation.

The deposition of zinc-zinc oxide nanoparticles (Zn-ZnO NPs) onto porous Ta2O5 surfaces enriched with calcium phosphate by DC magnetron sputtering was investigated to improve the surface antimicrobial activity without triggering an inflammatory response. Different sizes and amounts of Zn NPs obtained by two optimized different depositions and an additional thin carbon (C) layer deposited over the NPs were explored. The deposition of the Zn NPs and the C layer mitigates the surface porosity, increasing the surface hydrophobicity and decreasing the surface roughness. The possible antimicrobial effect and immune system activation of Zn-ZnO NPs were investigated in Candida albicans and macrophage cells, respectively. It was found that the developed surfaces displayed a fungistatic behavior, as they impair the growth of C. albicans between 5 and 24 h of culture. This behavior was more evident on the surfaces with bigger NPs and the highest amounts of Zn. The same trend was observed in both reactive oxygen species (ROS) generation and loss of C. albicans' membrane integrity. After 24 h of culture, cell toxicity was also dependent on the amount of the NPs. Cell toxicity was observed in surfaces with the highest amount of Zn NPs and with the C layer, while cells were able to grow without any signs of cytotoxicity in the porous surfaces with the lowest amount of NPs. The same Zn-dose-dependent behavior was noticed in the TNF-α production. The Zn-containing surfaces show a vastly inferior cytokine secretion than the lipopolysaccharide (LPS)-stimulated cells, indicating that the modified surfaces do not induce an inflammatory response from macrophage cells. This study provides insights for understanding the Zn amount threshold that allows a simultaneous inhibition of the fungi growth with no toxic effect and the main antimicrobial mechanisms of Zn-ZnO NPs, contributing to future clinical applications.

The role of the biomechanics analyst in swimming training and competition analysis.

Swimming analysts aid coaches and athletes in the decision-making by providing evidence-based recommendations. The aim of this narrative review was to report the best practices of swimming analysts that have been supporting high-performance athletes. It also aims to share how swimming analysts can translate applied research into practice. The role of the swimming analyst, as part of a holistic team supporting high-performance athletes, has been expanding and is needed to be distinguished from the job scope of a swimming researcher. As testing can be time-consuming, analysts must decide what to test and when to conduct the evaluation sessions. Swimming analysts engage in the modelling and forecast of the performance, that in short- and mid-term can help set races target-times, and in the long-term provide insights on talent and career development. Races can be analysed by manual, semi-automatic or fully automatic video analysis with single or multi-cameras set-ups. The qualitative and quantitative analyses of the swim strokes, start, turns, and finish are also part of the analyst job scope and associated with race performance goals. Land-based training is another task that can be assigned to analysts and aims to enhance the performance, prevent musculoskeletal injuries and monitor its risk factors.

The gold rush in para swimming: changes in the speed curve of a visually impaired world and paralympic champion - a case study.

This case study examined the association between 50 m freestyle and speed curve parameters of a world-class Paralympic swimmer and analysed the changes in speed curves and their frequency components across her performance levels. From 2018 to 2021, a visually impaired female swimmer (26.59 s in 50 m freestyle, S12 class) underwent 22 tests to obtain instantaneous speed synchronised with video recording. She regularly performed 50 m freestyle in competitions and time trials. The fast Fourier transformation method converted the speed signal into frequency domains and determined the relative contribution of the harmonics with two maxima and minima (H2, arms actions) and six maxima and minima (H6, legs actions). The functional paired t-test compared the speed curves at the beginning (PRE) and end (POST) of the analysed period. The 50 m freestyle time correlated with average speed (r = -0.50, p = 0.02). The contribution of H6 increased in the first year and remained large, whereas the contribution of H2 was lower throughout the whole period. POST was faster than PRE in five moments that coincide with the downward leg kick moments. These changes allowed her to stay longer at the upper part of the curve and improve performance over time.