Proteomics is advancing the understanding of stallion sperm biology.

The mammalian ejaculate is very well suited to proteomics studies. As such, research concerning sperm proteomics is offering a huge amount of new information on the biology of spermatozoa. Among domestic animals, horses represent a species of special interest, in which reproductive technologies and a sizeable market of genetic material have grown exponentially in the last decade. Studies using proteomic approaches have been conducted in recent years, showing that proteomics is a potent tool to dig into the biology of the stallion spermatozoa. The aim of this review is to present an overview of the research conducted, and how these studies have improved our knowledge of stallion sperm biology. The main outcomes of the research conducted so far have been an improved knowledge of metabolism, and its importance in sperm functions, the impact of different technologies on the sperm proteome, and the identification of potential biomarkers. Moreover, proteomics of seminal plasma and phosphoproteomics are identified as areas of major interest.

Habitat is more important than climate for structuring soil fungal communities associated in truffle sites.

The ectomycorrhizal fungi Tuber melanosporum Vittad. and Tuber aestivum Vittad. produce highly valuable truffles, but little is known about the soil fungal communities associated with these truffle species in places where they co-occur. Here, we compared soil fungal communities present in wild and planted truffle sites, in which T. melanosporum and T. aestivum coexist, in Mediterranean and temperate regions over three sampling seasons spanning from 2018 to 2019. We showed that soil fungal community composition and ectomycorrhizal species composition are driven by habitat type rather than climate regions. Also, we observed the influence of soil pH, organic matter content and C:N ratio structuring total and ectomycorrhizal fungal assemblages. Soil fungal communities in wild sites revealed more compositional variability than those of plantations. Greater soil fungal diversity was found in temperate compared to Mediterranean sites when considering all fungal guilds. Ectomycorrhizal diversity was significantly higher in wild sites compared to plantations. Greater mould abundance at wild sites than those on plantation was observed while tree species and seasonal effects were not significant predictors in fungal community structure. Our results suggested a strong influence of both ecosystem age and management on the fungal taxa composition in truffle habitats.

The future of equine semen analysis.

We are currently experiencing a period of rapid advancement in various areas of science and technology. The integration of high throughput 'omics' techniques with advanced biostatistics, and the help of artificial intelligence, is significantly impacting our understanding of sperm biology. These advances will have an appreciable impact on the practice of reproductive medicine in horses. This article provides a brief overview of recent advances in the field of spermatology and how they are changing assessment of sperm quality. This article is written from the authors' perspective, using the stallion as a model. We aim to portray a brief overview of the changes occurring in the assessment of sperm motility and kinematics, advances in flow cytometry, implementation of 'omics' technologies, and the use of artificial intelligence/self-learning in data analysis. We also briefly discuss how some of the advances can be readily available to the practitioner, through the implementation of 'on-farm' devices and telemedicine.

Spermatozoa cooled to 5°C one day after collection from porcine commercial semen doses retain sperm functionality with reduced bacterial load.

BACKGROUND: Commercial porcine semen is stored at 17°C, leading to a reduction of sperm quality and increase of bacterial growth. OBJECTIVES: To evaluate the effect of 5°C storage on porcine sperm functionality cooled one day after collection. MATERIALS AND METHODS: Semen doses (n = 40) were transported at 17°C and cooled at 5°C one day after collection. Spermatozoa were evaluated at Days 1, 4, and 7 for motility, viability, acrosome integrity, membrane stability, intracellular zinc, oxidative stress, and bacterial growth. RESULTS: Contaminated semen doses predominantly exhibited Serratia marcescens, with increasing bacterial load during 17°C storage. Under hypothermal storage, negative doses for bacteria growth at Day 1 remained negative, and bacterial load did not increase in bacterial contaminated samples. Motility was significantly reduced through 17°C storage, but at 5°C, motility was only reduced at Day 4. Samples with bacterial growth (35.0%, 14/40) had significantly reduced motility at 17°C, but motility was unaltered at 5°C. Plasma membrane and acrosome integrity without bacterial contamination were unaffected at 17°C, but were significantly reduced at 5°C on Day 7. Plasma membrane and acrosome integrity significantly decreased with bacterial contamination regardless of temperature. High mitochondrial activity in viable spermatozoa without bacteria was not altered by temperature, but was significantly reduced by bacterial contamination at 17°C. Membrane stability was significantly reduced at Day 4, but tended (p = 0.07) to be higher in samples without bacterial growth. Viable spermatozoa exhibiting high zinc were significantly reduced throughout storage regardless of temperature. Oxidative stress levels were not altered, but significantly increased with bacterial contamination at 17°C. DISCUSSION AND CONCLUSION: Porcine spermatozoa cooled to 5°C one day after collection retain functional attributes similar to spermatozoa stored at 17°C, but have a reduced bacterial load. Cooling extended boar semen to 5°C is feasible after transport to avoid modifying semen production.

The concentration of glucose in the media influences the susceptibility of stallion spermatozoa to ferroptosis.

In brief: Although common in many commercial extenders, supraphysiological concentrations of glucose in the media may be detrimental to stallion spermatozoa. In this study, we present evidence that these elevated glucose levels may predispose spermatozoa to ferroptosis. Abstract: Stallion spermatozoa depend on oxidative phosphorylation as their major source of ATP; however, the metabolism of these cells is complex and a great degree of metabolic plasticity exists. The composition of the media in which the spermatozoa are extended, or exposed to in the mare's reproductive tract, exerts a profound effect on sperm function and may even accelerate cell demise. Recent research indicates that high concentrations of glucose in the media, although common in commercial extenders, may be detrimental. To determine if supraphysiological glucose concentration may induce or predispose to ferroptosis (a caspase-independent form of programmed cell death, triggered by oxidative stress), stallion spermatozoa were incubated under different concentrations of glucose, 67 mM (HG) or 1 mM plus 10 mM pyruvate (LG-HP), in the presence or absence of known inductors of ferroptosis. Furthermore, we developed a single-cell flow metabolic assay to identify different metabolic phenotypes in spermatozoa. Storage and incubation of spermatozoa under high glucose concentrations led to an increase in the percentage of necrotic spermatozoa (P < 0.0001). Moreover, ferroptosis was induced more intensely in sperm in media with high glucose concentrations (P < 0.0001). Finally, we observed that induction of ferroptosis modified two proteins (oxoglutarate dehydrogenase and superoxide dismutase 2) in spermatozoa incubated in media containing 67 mM glucose but not in media containing 1 mM glucose and 10 mM pyruvate. The composition of the media, especially the concentration of glucose, exerts a major impact on the functionality and life span of the spermatozoa. The results reported here may pave the way for improvements in existing semen extenders.

Pyruvate enhances stallion sperm function in high glucose media improving overall metabolic efficiency.

If a mechanism of more efficient glycolysis depending on pyruvate is present in stallion spermatozoa, detrimental effects of higher glucose concentrations that are common in current commercial extenders could be counteracted. To test this hypothesis, spermatozoa were incubated in a 67 mM Glucose modified Tyrode's media in the presence of 1- or 10-mM pyruvate and in the Tyrode's basal media which contains 5 mM glucose. Spermatozoa incubated for 3 h at 37 °C in 67 mM Tyrode's media with 10 mM pyruvate showed increased motility in comparison with aliquots incubated in Tyrode's 5 mM glucose and Tyrode's 67 mM glucose (57.1 ± 3.5 and 58.1 ± 1.9 to 73.0 ± 1.1 %; P < 0.01). Spermatozoa incubated in Tyrode's with 67 mM glucose 10 mM pyruvate maintained the viability along the incubation (64.03 ± 15.4 vs 61.3 ± 10.2), while spermatozoa incubated in 67 mM Glucose-Tyrode's showed a decrease in viability (38.01 ± 11.2, P < 0.01). 40 mM oxamate, an inhibitor of the lactate dehydrogenase LDH, reduced sperm viability (P < 0.05, from 76 ± 5 in 67 mM Glucose/10 mM pyruvate to 68.0 ± 4.3 %, P < 0.05). Apoptotic markers increased in the presence of oxamate. (P < 0.01). UHPLC/MS/MS showed that 10 mM pyruvate increased pyruvate, lactate, ATP and NAD+ while phosphoenolpyruvate decreased. The mechanisms that explain the improvement of in presence of 10 mM pyruvate involve the conversion of lactate to pyruvate and increased NAD+ enhancing the efficiency of the glycolysis.

Intraocular pressure changes under an atmospheric pressure spectrum in a multiplace hyperbaric chamber / Análise de alterações da pressão intraocular (PIO) sob o espectro da pressão atmosférica em uma câmara hiperbárica multi-lugar

ABSTRACT Purpose: To evaluate the influence of atmospheric pressure changes on the behavior of intraocular pressure of healthy military individuals-students and instructors of the National Navy's Diving & Rescue School at the "ARC BOLÍVAR" naval base-during a simulated immersion in the hyperbaric chamber of the Naval Hospital of Cartagena. Methods: A descriptive exploratory study was performed. The intraocular pressure was measured at different atmospheric pressures during 60-min sessions in the hyperbaric chamber while breathing compressed air. The maximum simulated depth was 60 feet. Participants were students and instructors of the Naval Base's Diving and Rescue Department. Results: A total of 48 eyes from 24 divers were studied, of which 22 (91.7%) were male. The mean age of the participants was 30.6 (SD=5.5) years, ranging from 23 to 40. No participant had a history of glaucoma or ocular hypertension. The mean base intraocular pressure at sea level was 14 mmHg, which decreased to 13.1 mmHg (decreased by 1.2 mmHg) at 60 feet deep (p=0.0012). However, during the safety stop at 30 feet, the mean IOP kept decreasing until reaching 11.9 mmHg (p<0.001). By the end of the session, the mean intraocular pressure reached 13.1 mmHg, which is inferior and statistically significant when compared with the intraocular pressure base mean (p=0.012). Conclusions: In healthy individuals, the intraocular pressure decreases when reaching a depth of 60 feet (2.8 absolute atmosphere pressure) and it decreases even more during ascension at 30 feet. Measurements at both points were significantly different when compared with base intraocular pressure. The final intraocular pressure was lower than the baseline intraocular pressure, suggesting a residual and prolonged effect of the atmospheric pressure on intraocular pressure.

RESUMO Objetivo: Avaliar a influência das alterações da pressão atmosférica no comportamento da pressão intraocular de indivíduos militares saudáveis-alunos e instrutores da Escola de Mergulho e Resgate da Marinha Nacional na base naval "ARC BOLÍVAR"-durante uma imersão simulada na câmara hiperbárica do Hospital da Marinha de Cartagena. Métodos: Realizamos um estudo exploratório descritivo. A pressão intraocular foi medida em diferentes pressões atmosféricas durante sessões de 60 minutos na câmara hiperbárica respirando ar comprimido. A profundidade máxima simulada foi de 60 pés. Os participantes eram alunos e instrutores do Departamento de Mergulho e Resgate da Base Naval. Resultados: Quarenta e oito olhos de 24 mergulhadores foram estudados. Vinte e dois participantes (91,7%) eram do sexo masculino. A média de idade dos participantes foi de 30,6 (DP=5,5) anos, variando de 23 a 40. Nenhum participante tinha histórico de glaucoma ou hipertensão ocular. A média de base da pressão intraocular ao nível do mar foi de 14 mmHg, diminuindo para 13,1 mmHg (queda de 1,2 mmHg) a 60 pés de profundidade (p=0,0012). Entretanto, durante a parada de segurança a 30 pés, a pressão intraocular média continuou diminuindo até atingir 11,9 mmHg (p<0,001). Ao final da sessão, a pressão intraocular média atingiu 13,1 mmHg, valor inferior e estatisticamente significativo quando comparada à média de base da pressão intraocular (p=0,012). Conclusões: Em indivíduos saudáveis, a pressão intraocular diminui ao atingir uma profundidade de 60 pés (2,8 de pressão atmosférica absoluta) e diminui ainda mais durante a ascensão a 30 pés. As medidas em ambos os pontos foram significativamente diferentes quando comparadas à pressão intraocular de base. A pressão intraocular final foi menor do que a pressão intraocular de base, sugerindo um efeito residual e prolongado da pressão atmosférica sobre a pressão intraocular.

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose-10 mm pyruvate-based media.

BACKGROUND: Most commerce of equine seminal doses is carried out using commercial extenders under refrigeration at 5°C. OBJECTIVES: To determine if 10 mm pyruvate in a 67 mm glucose extender and storage at 22°C could be the basis of an alternative storage method to cooling to 5°C. MATERIAL AND METHODS: Stallion ejaculates were extendedin: INRA96 (67 mm glucose, non-pyruvate control), modified Tyrode's (67 mm glucose-10 mm pyruvate), supplemented with 0, 10, 50, and 100 µM itaconate. As itaconate was vehiculated in DMSO, a control vehicle was also included. Sperm motility, viability, mitochondrial membrane potential, and production of reactive oxygen species were measured after collection and again after 48 and 96 h of storage at 22°C. To disclose molecular metabolic changes, spermatozoa were incubated up to 3 h in modified Tyrode's 67 mm glucose-10 mm pyruvate and modified Tyrode's 67 mm glucose, and metabolic analysis conducted. RESULTS: After 96 h of storage aliquots stored in the control, INRA96 had a very poor total motility of 5.6% ± 2.3%, while in the 67 mm glucose-10 mm pyruvate/10 µm itaconate extender, total motility was 34.7% ± 3.8% (p = 0.0066). After 96 h, viability was better in most pyruvate-based media, and the mitochondrial membrane potential in spermatozoa extended in INRA96 was relatively lower (p < 0.0001). Metabolomics revealed that in the spermatozoa incubated in the high pyruvate media, there was an increase in the relative amounts of NAD+ , pyruvate, lactate, and ATP. DISCUSSION AND CONCLUSIONS: Aliquots stored in a 67 mm glucose-10 mm pyruvate-based medium supplemented with 10 µM itaconate, maintained a 35% total motility after 96 h of storage at 22°C, which is considered the minimum acceptable motility for commercialization. Improvements may be related to the conversion of pyruvate to lactate and regeneration of NAD+ .

Foot and Ankle Bone Marrow Edema Assessment in Long Distance Runners.

BACKGROUND: The clinical significance of foot and ankle bone marrow edema (BME) is poorly understood. METHODS: Magnetic resonance imaging (MRI) and the visual analog scale (VAS) pain scores were collected for 17 runners who participated in the Twin Cities Marathon, immediately postmarathon and at a 6-week follow-up. Bone marrow edema lesions were classified using lesion frequency, anatomical location, and grading scale change to calculate a BME score for each affected bone. Spearman rank correlation coefficient test was used to identify a possible correlation between VAS and postmarathon BME. A paired Student t test was used to detect differences between total mileage ran 6 weeks postmarathon in participants with or without BME. RESULTS: After completing the marathon, 8 BME lesions were identified in 5 participants (29.4%; 5/17), 3 were men, and 2 were women, with a mean age of 33.8 years (range: 24-52), and BMI of 22.9 ± 4. All lesions were resolved on 6-week follow-up imaging. VAS pain scores did not correlate with postmarathon BME. A significant difference in total miles logged over 6 weeks postmarathon could not be found among participants with and without BME. CONCLUSION: Foot and ankle BME changes identified by MRI were not correlated to clinical symptoms and may resolve with self-directed activity in less time than other areas of the lower extremity. LEVEL OF EVIDENCE: Level II, Prospective Cohort Study.

Adaptación transcultural y validación de la versión en español del cuestionario de carga de tratamiento en personas con diabetes mellitus tipo 2 que concurren a centros de atención primaria del sector público de salud. San Juan / Transcultural adaptation and validation of the spanish version of the treatment burden questionnaire for people with diabetes mellitus type 2 that attend to primary health care centers within the public health care system. San Juan

Introducción: el Treatment Burden Questionnaire (TBQ) evalúa la carga que implica el autocuidado de personas con enfermedades crónicas y el impacto que la asistencia sanitaria genera en su calidad de vida. Objetivos: adaptar y validar psicométricamente la versión en español del cuestionario TBQ-esclerosis múltiple al contexto de la diabetes mellitus (DM) en pacientes atendidos en el sector público de Argentina. Materiales y métodos: estudio analítico de adaptación transcultural y validación psicométrica de un cuestionario. Se realizó un análisis factorial exploratorio (AFE) y un análisis factorial confirmatorio (AFC). La validez de criterio concurrente se evaluó a través de la correlación Pearson con tres escalas (OMS-5, PHQ-9 y test de Morisky-Green). La fiabilidad se analizó a través del coeficiente alfa de Cronbach, de la fiabilidad compuesta y la varianza media extractada. Resultados: participaron 256 personas. La versión adaptada fue adecuadamente comprendida por la población destinataria. El AFE (n=100) permitió hipotetizar cuatro dominios: 1) tratamiento farmacológico; 2) asistencia sanitaria/sistema de salud; 3) mantenimiento de un estilo de vida y su repercusión económica; 4) esfera psicosocial. En el AFC (n=156) las cargas factoriales del 87,5% (14/16) de los ítems fueron mayores a 0,5 con significancia estadística. Se documentó correlación inversa (-0,46; p=0,0002) con el puntaje OMS-5 y directa (0,36; p=0,0046) con PHQ-9. Los pacientes categorizados como "no adherentes" mediante la escala de Morisky-Green-Levine arrojaron un resultado de TBQ-DM de 16,99 (IC 95%: 0,95 a 33,03) puntos más que los pacientes categorizados como "adherentes" (p=0,0383). La consistencia interna del cuestionario fue muy buena (alfa de Cronbach: 0,87), su fiabilidad compuesta fue aceptable y su validez divergente, baja. Conclusiones: el TBQ en español adaptado a DM2 tiene aceptables propiedades psicométricas. Su implementación podría ofrecer cuidados basados en la persona.

Introduction: the Treatment Burden Questionnaire (TBQ) assess the load that implies the self-care for people with chronic diseases and the impact that medical assistance produce in their quality of life. Objectives: to adapt and psychometrically validate the spanish version of the TBQ-multiple sclerosis questionnaire to the diabetes mellitus (DM) context in patients treated in the Argentinian public health sector. Materials and methods: analytic study of transcultural adaptation and psychometric validation of a questionnaire. An exploratory factor analysis (EFA) and a confirmatory factor analysis (CFA) were performed. Concurrent criterion validity was evaluated through Pearson correlation with three scales (WHO-5, PHQ-9, Morisky-Green Test). Reliability was evaluated through Cronbach's alpha coefficient, composite reliability, and average variance extracted. Results: 256 people participated. The adapted version was adequately understood by the target population. EFA (n=100) allowed for the hypothesis of four domains: 1) pharmacological treatment; 2) healthcare/health system; 3) maintenance of a lifestyle and its economic impact; 4) psychosocial sphere. In the CFA (n=156), the factor loadings of 87.5% (14/16) of the items were greater than 0.5 with statistical significance. An inverse correlation (-0.46; p=0.0002) with the WHO-5 index and a direct correlation (0.36; p=0.0046) with the PHQ-9 scale were documented. Patients categorized as "non-adherent" by the Morisky-Green-Levine scale had a TBQ-DM score of 16.99 (95% CI: 0.95 to 33.03) points higher than patients categorized as "adherent" (p=0.0383). The internal consistency of the questionnaire was very good (Cronbach's alpha: 0.87), its composite reliability was acceptable, and its divergent validity was low. Conclusions: the spanish TBQ adapted to DM2 has acceptable pyshcometric properties. Its implementation can help offer a person-centered care.

Exploiting the PIR Sensor Analog Behavior as Thermoreceptor: Movement Direction Classification Based on Spiking Neurons.

Pyroelectric infrared sensors (PIR) are widely used as infrared (IR) detectors due to their basic implementation, low cost, low power, and performance. Combined with a Fresnel lens, they can be used as a binary detector in applications of presence and motion control. Furthermore, due to their features, they can be used in autonomous intelligent devices or included in robotics applications or sensor networks. In this work, two neural processing architectures are presented: (1) an analog processing approach to achieve the behavior of a presynaptic neuron from a PIR sensor. An analog circuit similar to the leaky integrate and fire model is implemented to be able to generate spiking rates proportional to the IR stimuli received at a PIR sensor. (2) An embedded postsynaptic neuron where a spiking neural network matrix together with an algorithm based on digital processing techniques is introduced. This structure allows connecting a set of sensors to the post-synaptic circuit emulating an optic nerve. As a case study, the entire neural processing approach presented in this paper is applied to optical flow detection considering a four-PIR array as input. The results validate both the spiking approach for an analog sensor presented and the ability to retrieve the analog information sent as spike trains in a simulated optic nerve.

Proteomic research on new urinary biomarkers of renal disease in canine leishmaniosis: Survival and monitoring response to treatment.

The objective of our study was to search for survival biomarkers (SB) and treatment response monitoring biomarkers (TRMB) in the urinary proteome of dogs with renal disease secondary to canine leishmaniosis (CanL), using UHPLC-MS/MS. The proteomic data are available via ProteomeXchange with identifier PXD042578. Initially, a group of 12 dogs was evaluated and divided into survivors (SG; n = 6) and nonsurvivors (NSG; n = 6). A total of 972 proteins were obtained from the evaluated samples. Then, bioinformatic analysis reduced them to 6 proteins like potential SB increased in the NSG, specifically, Haemoglobin subunit Alpha 1, Complement Factor I, Complement C5, Fibrinogen beta chain (fragment), Peptidase S1 domain-containing protein, and Fibrinogen gamma chain. Afterwards, SG was used to search for TRMB, studying their urine at 0, 30, and 90 days, and 9 proteins that decreased after treatment were obtained: Apolipoprotein E, Cathepsin B, Cystatin B, Cystatin-C-like, Lysozyme, Monocyte differentiation CD14, Pancreatitis-associated precursor protein, Profilin, and Protein FAM3C. Finally, enrichment analysis provided information about the biological mechanisms in which these proteins are involved. In conclusion, this study provides 15 new candidate urinary biomarkers and an improved understanding of the pathogenesis of kidney disease in CanL.

Ovine fertility by artificial insemination in the breeding season could be affected by intraseasonal variations in ram sperm proteomic profile.

It is important to note that seasonality could affect ram reproductive parameters, and therefore, fertility results after artificial insemination. In this work, 1) we assessed fertility rates after cervical artificial insemination of 11,805 ewes at the beginning (June 21st to July 20th) and at the end (November 20th to December 21st) of the reproductive season in the Assaf breed for the last four years, and 2) we aimed to identify male factors influencing the different reproductive success obtained depending on the time at the mating season in which ovine artificial insemination was performed. For this purpose, we evaluated certain ram reproductive and ultrasonographical parameters as well as we performed a multiparametric and proteomic sperm analysis of 6-19 rams at two very distant points in the mating season (July as Early Breeding Season -EBS- and November as Late Breeding Season -LBS-). Rutinary assessments carried out in the ovine reproduction centers (testicular volume, libido, sperm production and mass motility) showed non-significant differences (P ≥ 0.05) between both studied times, as well as the ram ultrasonographic evaluation (Resistive and Pulsatility Index as Doppler parameters; and pixels mean gray level, and hypoechoic areas percentage and density as echotexture parameters). However, at level of sperm functionality, although sperm quality appeared non-significantly lower (P ≥ 0.05) in the EBS, we identified a significantly different (P < 0.05) sperm proteomic profile between the seasonality points. The following proteins were identified with the lowest abundance in the EBS with a fold change > 4, a P = 2.40e-07, and a q = 2.23e-06: Fibrous Sheath-Interacting Protein 2, Disintegrin and Metalloproteinase Domain-Containing Protein 20-like, Phosphoinositide-Specific Phospholipase C, Tektin 5, Armadillo Repeat-Containing Protein 12 Isoform X3, Solute Carrier Family 9B1, Radial Spoke Head Protein 3 Homolog, Pro-Interleukin-16, NADH Dehydrogenase [Ubiquinone] 1 Alpha Subcomplex Subunit 8, Testis, Prostate and Placenta-Expressed Protein, and Acyl Carrier Protein Mitochondrial. In conclusion, while our basic analyses on male and sperm quality showed similar results between the beginning and the end of the breeding season, on a proteomic level we detected a lower expression of sperm proteins linked to the energy metabolism, sperm-oocyte interactions, and flagellum structure in the EBS. Probably, this different protein expression could be related to the lower fertility rate of Assaf ewes after cervical artificial insemination at this time. More importantly, sperm proteins can be used as highly effective molecular markers in predicting sperm fertilization ability related to intraseasonal variations.

Intraocular pressure changes under an atmospheric pressure spectrum in a multiplace hyperbaric chamber.

PURPOSE: To evaluate the influence of atmospheric pressure changes on the behavior of intraocular pressure of healthy military individuals-students and instructors of the National Navy's Diving & Rescue School at the "ARC BOLÍVAR" naval base-during a simulated immersion in the hyperbaric chamber of the Naval Hospital of Cartagena. METHODS: A descriptive exploratory study was performed. The intraocular pressure was measured at different atmospheric pressures during 60-min sessions in the hyperbaric chamber while breathing compressed air. The maximum simulated depth was 60 feet. Participants were students and instructors of the Naval Base's Diving and Rescue Department. RESULTS: A total of 48 eyes from 24 divers were studied, of which 22 (91.7%) were male. The mean age of the participants was 30.6 (SD=5.5) years, ranging from 23 to 40. No participant had a history of glaucoma or ocular hypertension. The mean base intraocular pressure at sea level was 14 mmHg, which decreased to 13.1 mmHg (decreased by 1.2 mmHg) at 60 feet deep (p=0.0012). However, during the safety stop at 30 feet, the mean IOP kept decreasing until reaching 11.9 mmHg (p<0.001). By the end of the session, the mean intraocular pressure reached 13.1 mmHg, which is inferior and statistically significant when compared with the intraocular pressure base mean (p=0.012). CONCLUSIONS: In healthy individuals, the intraocular pressure decreases when reaching a depth of 60 feet (2.8 absolute atmosphere pressure) and it decreases even more during ascension at 30 feet. Measurements at both points were significantly different when compared with base intraocular pressure. The final intraocular pressure was lower than the baseline intraocular pressure, suggesting a residual and prolonged effect of the atmospheric pressure on intraocular pressure.

Smart Shoe Insole Based on Polydimethylsiloxane Composite Capacitive Sensors.

Nowadays, the study of the gait by analyzing the distribution of plantar pressure is a well-established technique. The use of intelligent insoles allows real-time monitoring of the user. Thus, collecting and analyzing information is a more accurate process than consultations in so-called gait laboratories. Most of the previous published studies consider the composition and operation of these insoles based on resistive sensors. However, the use of capacitive sensors could provide better results, in terms of linear behavior under the pressure exerted. This behavior depends on the properties of the dielectric used. In this work, the design and implementation of an intelligent plantar insole composed of capacitive sensors is proposed. The dielectric used is a polydimethylsiloxane (PDMS)-based composition. The sensorized plantar insole developed achieves its purpose as a tool for collecting pressure in different areas of the sole of the foot. The fundamentals and details of the composition, manufacture, and implementation of the insole and the system used to collect data, as well as the data samples, are shown. Finally, a comparison of the behavior of both insoles, resistive and capacitive sensor-equipped, is made. The prototype presented lays the foundation for the development of a tool to support the diagnosis of gait abnormalities.

The Autophagy Marker LC3 Is Processed during the Sperm Capacitation and the Acrosome Reaction and Translocates to the Acrosome Where It Colocalizes with the Acrosomal Membranes in Horse Spermatozoa.

Despite its importance in somatic cells and during spermatogenesis, little is known about the role that autophagy may play in ejaculated spermatozoa. Our aim was to investigate whether the molecular components of autophagy, such as microtubule-associated protein 1 light chain 3 (LC3), are activated in stallion spermatozoa during the capacitation and acrosome reaction and if this activation could modulate these biological processes. To analyze the autophagy turnover, LC3I and LC3II proteins were assessed by western blotting, and the ratio between both proteins (LC3II/LC3I) was calculated. In somatic cells, this ratio indicates that autophagy has been activated and similar LC3 processing has been described in mammalian spermatozoa. The subcellular localization of autophagy-related proteins was assessed by immunofluorescence with specific antibodies that recognized Atg16, Beclin-1, and LC3. The colocalization of acrosomal membranes (PNA) and LC3 was studied by confocal microcopy, and the acrosome reacted cells were quantified by flow cytometry. The incubation of stallion sperm in capacitating conditions (BWW; 3 h) significantly increased LC3 processing. This increment was three to four times higher after the induction of the acrosome reaction in these cells. LC3 was mainly expressed in the head in mature ejaculated sperm showing a clear redistribution from the post-acrosomal region to the acrosome upon the incubation of sperm in capacitating conditions (BWW, 3 h). After the induction of the acrosome reaction, LC3 colocalized with the acrosome or the apical plasmalemma membranes in the head of the stallion spermatozoa. The inhibition or activation of autophagy-related pathways in the presence of autophagy activators (STF-62247) or inhibitors (E-64d, chloroquine) significantly increased LC3 processing and increased the percent of acrosome reacted cells, whereas 3-methyladenine almost completely inhibited LC3 processing and the acrosome reaction. In conclusion, we found that sperm capacitation and acrosome reaction could be regulated by autophagy components in sperm cells ex vivo by processes that might be independent of the intraluminal pH of the acrosome and dependent of LC3 lipidation. It can be speculated that, in stallion sperm, a form of noncanonical autophagy utilizes some components of autophagy machinery to facilitate the acrosome reaction.

Phosphoproteomics for the identification of new mechanisms of cryodamage: the role of SPATA18 in the control of stallion sperm function†.

Although recent research has addressed the impact of cryopreservation on the stallion sperm proteome, studies addressing the stallion sperm phosphoproteome are lacking. In the present study, the data set of proteomes of fresh and cryopreserved spermatozoa were reanalyzed, showing that cryopreservation caused significant changes in the phosphoproteome. The phosphoproteins reduced most significantly by cryopreservation were Ca2+binding tyrosine phosphorylation regulated, protein kinase cAMP-activated catalytic subunit beta (CABYR), mitochondria eating protein (SPATA18), A kinase anchoring protein 4 (AKAP4), A-kinase anchoring protein 3 (AKAP3) and the Family with sequence similarity 71 member B (FAM71B). These proteins belong to the gene ontology (GO) terms sperm fibrous sheath (GO: 0035686), and sperm principal piece (GO: 0097228). The regulatory interactions between kinases and phosphorylation sites on the proteins that were affected most were also investigated, and the potential kinases (based on human orthologs) involved in the regulation of these phosphoproteins identified were: PKCß for SPATA18 and GSK3ß for CABYR. Kinase inhibition assays were also conducted showing that kinases phosphorylating the above-mentioned proteins play an important role in their activity and thus, phosphorylation controls the activity of these proteins and their role in the regulation of the functionality and viability of stallion spermatozoa. In conclusion, the data reported here contribute to the understanding of the fact that the dephosphorylation of certain proteins is a molecular lesion induced by cryopreservation in the stallion spermatozoa.

Validity and reliability of the Spanish version of the ACE tool for assessing competencies in evidence-based medicine in medical students.

OBJECTIVE: To translate and culturally adapt the tool 'Assessing Competency in evidence-based medicine (EBM)' (ACE) to Spanish and to implement it in a cohort of medical students for the evaluation of the instrument's psychometric properties. DESIGN: Bilingual translators produced a translation and backtranslation of the original instrument, with interim consensus in each stage with oversight and input by a group of experts. We then performed cognitive interviews to adapt the wording of the tool culturally. Finally, we implemented the final version in a cohort of medical students on a virtual general practice course with EBM modules. SETTING: Medical School in Buenos Aires, Argentina. Due to restrictions to in-person teaching during the COVID-19 pandemic, we conducted this study in the context of virtual learning. PARTICIPANTS: We included 125 fourth and fifth-year medical students. MAIN OUTCOME MEASURES: We measured internal consistency with the Kuder-Richardson coefficient (>0.6 as a threshold for reliability) and construct validity through a Pearson's correlation between the examinations carried out with the translated instrument and the results of the regular examinations of EBM in the same students (expected values of at least 0.3 to 0.7). We also compared the total score of the instrument of fifth-year students to fourth-year students. RESULTS: As for measurements for internal consistency, the coefficient Kuder-Richardson resulted in a value of 0.268, below our prespecified threshold. For construct validity, the Pearson correlation between the sum of the items and regular examinations was 0.139, also below our prespecified threshold. However, fifth-year students averaged 0.94 points more than fourth-year students (95% CI 0.24 more to 1.65 more). CONCLUSION: The translated and cross-culturally adapted version of the ACE tool into Spanish had low reliability and validity in an MBE course taught and evaluated in a virtual environment. TRIAL REGISTRATION: Not applicable.