Proliferation and apoptosis dynamics of the normal canine mammary gland during the oestrous cycle evaluated by Ki-67 and Caspase-3 indexes.

Understanding the normal physiology of the canine mammary gland (CMG) is crucial, as it provides a foundational reference for understanding canine mammary neoplasms. The relation between the Proliferation Index (PI) indicated by Ki-67 expression, along with the Apoptotic Index (AI) determined through Caspase-3 expression during the oestrous cycle, is inadequately documented in existing literature. This study seeks to offer insights into the interplay between PI and AI in the CMG across oestrous cycle phases. An extensive investigation was conducted on a diverse case series of bitches (n = 18). Oestrous cycle stages were determined through vaginal cytology, histological examination of the reproductive tract and serum progesterone and oestradiol concentrations. The entire mammary chain was histologically examined, and proliferation and apoptosis were assessed via double immunohistochemistry employing anti-Ki-67 and Caspase-3 antibodies. PI and AI were evaluated through a systematic random sampling approach, counting a minimum of 200 cells for each cell type. There was a significantly higher PI during early dioestrus in all mammary gland components, with a greater proportion of positive cells observed in epithelial cells compared to stromal cells. The highest PI was detected in epithelial cells within the end buds. Significant differences were found in Ki-67 labelling across the cranial mammary glands. A positive and strong correlation was noted between progesterone concentration and PI in epithelial cells. The AI remained consistently low throughout the oestrous cycle, with few differences observed across histological components. Caspase-3 labelling displayed the highest positivity in caudal mammary pairs. A negative and moderate correlation was identified between progesterone concentration and AI in interlobular mesenchymal cells. This study highlights the influence of endocrine regulation on cell proliferation indices in mammary tissue, emphasizing the need to consider these hormonal variations in toxicopathological studies involving canine mammary gland.

Psychological Aspects of Motocross Racing Considering Expected, Perceived, and Actual Performance.

Motocross racing is a seldom-researched popular extreme sport. This field research aimed to investigate feeling states, perceived arousal, anxiety, and negative and positive affect in the anticipatory and recovery race periods and their relationship to expected and perceived performance. Twenty Motocross racers completed psychometric scales before and after a national championship race. Results revealed that objective performance was unrelated to psychological measures. Arousal, anxiety, and positive affect were lower after the race. Expected performance was unrelated to postrace measures. Still, perceived performance correlated significantly with the feeling state, anxiety, and positive affect after the race and the feeling state before the race. Furthermore, racers who performed as expected or better showed improved feeling states after the race compared with those who did worse than expected. The core affect of the latter group declined. This research on psychological states during Motocross races could motivate new initiatives for future studies.

An immersive simulation strategy to teach canine vaginal cytology: A multi-centre study.

Vaginal cytology (VC) is an essential technique for monitoring the bitch's estrus cycle. Currently, animal-free teaching methodologies have not been investigated for VC. Hence, this study aimed to evaluate an immersive simulation with a VC model and augmented reality tools. Students (n = 219) from four universities were enrolled, having learning stations with models for practising VC that provided immediate feedback on the technique. Augmented reality tools comprised QR codes that endorsed students to short videos of owners' avatars reporting the clinical reproductive story of the simulated animals and slides with QR codes leading to microscopy slide navigation videos. Proestrus, estrus, diestrus, anestrus and vaginitis were identified in the learning stations. The students' perceptions were evaluated through questionnaires assessing satisfaction, motivation, confidence, impact on learning and diagnostic accuracy. Before the immersive simulation, students had no experience with VC, being afraid of doing a VC with a live dog. Almost all the students considered practicing VC as essential and 94% reported that repeating the procedure (>2 times) was the most important parameter for learning. The simulation activity lasted ≈3 h and significantly improved the confidence of students, being less afraid of doing a VC in a live animal. Slide navigation videos improved the diagnostic accuracy of the estrus cycle stage, and students diagnosed the estrus and vaginitis cases more accurately. The immersive simulation strategy allowed repeated practice in a safe, motivated and standardized environment, being appraised by students as an essential strategy for learning VC.

Development and Validation of a Simulation Model for Collection of Canine Vaginal Samples.

Vaginal cytology is a widely used cytological technique mostly taught by observation, either through direct tutoring or videos. To the best of our knowledge, vaginal cytology simulators have never been assessed in veterinary medicine. Twenty-five undergraduate students with no prior experience in canine vaginal sampling were randomly assigned to two groups that either practiced the procedure in a simulator or a live animal. An inverted classroom design was followed. After observing a video tutorial, students practiced with the simulator/live animal for two classes. Three weeks later, they performed a vaginal cytology on a live animal being recorded. The videos were evaluated through an objective structured clinical examination (OSCE) by an observer blinded to the student's groups. The learning outcome was compared through OSCE pass rates and questionnaires. The simulation model was made by 3D printing and soft silicone for the vulvar labia, having pink and blue colored vaseline in the correct and incorrect locations for sampling. The model was economic and accurately replicated the female reproductive tract. It provided immediate feedback to students, by obtaining pink or blue swabs from the correct and incorrect locations, respectively. Students reported that three to five or more attempts were needed to properly learn the procedure, thus justifying the need for a simulator. No differences in the OSCE pass rates were observed between the groups. The simulation model was effective for learning the vaginal cytology procedure, replacing the use of live animals. This low-cost model should be incorporated in the tool-kit of reproduction classes.

The Use of Simulators for Teaching Fine Needle Aspiration Cytology in Veterinary Medicine.

Fine needle aspiration (FNA) is widely used by veterinary practitioners, being taught mostly by observation. Simulators are known to enhance students' learning of practice skills, but to our knowledge, FNA simulators have never been assessed in veterinary medicine. Fifty-one undergraduate students with no prior experience in cytology were randomly assigned to two groups that practiced on either a box simulator (with artificial nodules) or a fruit (banana). An in-class flip was followed, in which students first observed a FNA video tutorial and then used their assigned simulator for 15 minutes maximum. Students then attempted a FNA on an animal model and were evaluated through an objective structured clinical examination (OSCE). Learning outcomes of each model was compared through questionnaires, OSCE pass rates, and quality of produced smears. After observing the video, no student reported being able to conduct a FNA on a live animal, whereas most assured that they would be able to do so after using a simulator. Students practiced more on the box model (14.8 ± 0.8 min) than on the fruit (8.5 ± 2.2 min). At evaluation, students who had practiced on the box had more puncturing accuracy than those who had practiced on the fruit. Still, no differences in OSCE pass rates existed. Simulation models thus were effective for learning FNA, but the box simulator seemed to be more successful than the fruit in terms of deliberate practice. This appears to have a positive effect on students' puncturing accuracy, which has clinical relevance.

A Brazilian male with typical oral and pulmonary paracoccidioidomycosis.

An autochthonous case of paracoccidioidomycosis was reported in a city of north Iran. This condition is a well-known endemic fungal infection highly prevalent in Latin American countries, with an incidence of 1 to 3.7 cases per 100.000 annually in Brazil. The classical features are cutaneous lesions, lymph node, and pulmonary involvements, while typical oral changes are superficial ulcers with hemorrhage and moriform aspect. Herein is reported an adult male patient with characteristic oral and pulmonary lesions. Rural environment, male gender, cigarette smoking, and alcohol abuse were risk factors; and clinical history, imaging studies, and histopathologic data established the diagnosis. The patient improved well by administration of sulfamethoxazole plus trimethoprim. The aim of this case study is to enhance the awareness of generalists about this mycosis.

A utilização de simuladores para o ensino da punção de agulha fina em medicina veterinária.

A punção de agulha fina (PAF) é amplamente utilizada por veterinários, sendo ensinada maioritariamente por observação. Atualmente, considera-se que a utilização de simuladores melhora a aprendizagem de procedimentos práticos. No entanto, simuladores de PAF ainda não foram avaliados em Medicina Veterinária. Cinquenta e um estudantes de Veterinária sem experiência prévia em Citologia foram distribuídos aleatoriamente em dois grupos que usaram um simulador (caixa com nódulos artificiais) e uma peça de fruta (banana). Foi utilizado um desenho de aula invertida com estações: primeiramente, os estudantes observaram um vídeo tutorial sobre a PAF, utilizando em seguida o simulador ou a peça de fruta durante um máximo de 15 minutos. Depois, os estudantes efetuaram o procedimento num modelo animal realista, realizando-se uma avaliação clínica objetiva estruturada (ACOE). A aprendizagem através dos modelos foi comparada por meio de questionários, taxa de aprovação em ACOE e qualidade dos esfregaços obtidos. Após observar o vídeo tutorial, nenhum estudante manifestou ser capaz de fazer a PAF num animal vivo. Por oposição, a maioria revelou ser capaz após a simulação. Os estudantes praticaram mais tempo na caixa (14,8 ± 0,8 min) do que na peça de fruta (8,5 ± 2,2 min). Na avaliação, os primeiros tinham maior precisão na PAF. Ainda assim, não existiram diferenças na taxa de aprovação em ACOE. Portanto, os dois modelos são eficazes para a aprendizagem da PAF, mas a caixa tem vantagens quanto à repetição autónoma da prática. Essa repetição parece ter efeitos positivos na precisão da PAF, o que tem relevância do ponto de vista clínico.

Long live the liver: immunohistochemical and stereological study of hepatocytes, liver sinusoidal endothelial cells, Kupffer cells and hepatic stellate cells of male and female rats throughout ageing.

Male/female differences in enzyme activity and gene expression in the liver are known to be attenuated with ageing. Nevertheless, the effect of ageing on liver structure and quantitative cell morphology remains unknown. Male and female Wistar rats aged 2, 6, 12 and 18 months were examined by means of stereological techniques and immunohistochemical tagging of hepatocytes (HEP), liver sinusoidal endothelial cells (LSEC), Kupffer cells (KC) and hepatic stellate cells (HSC) in order to assess the total number and number per gram of these cells throughout life. The mean cell volume of HEP and HSC, the lobular position and the collagen content of the liver were also evaluated with stereological techniques. The number per gram of HSC was similar for both genders and was maintained throughout ageing. The mean volume of HSC was also conserved but differences in the cell body and lobular location were observed. Statistically significant gender differences in HEP were noted in young rats (females had smaller and more binucleated HEP) but were attenuated with ageing. The same occurred for KC and LSEC, since the higher number per gram in young females disappeared in older animals. Liver collagen increased with ageing but only in males. Thus, the numbers of these four cell types are related throughout ageing, with well-defined cell ratios. The shape and lobular position of HSC change with ageing in both males and females. Gender dimorphism in HEP, KC and LSEC of young rat liver disappears with ageing.

Multiple evolutionary units and demographic stability during the last glacial maximum in the Scytalopus speluncae complex (Aves: Rhinocryptidae).

The Atlantic Forest (AF) of South America harbors one of the world's highest bird species richness, but to date there is a deficient understanding of the spatial patterns of genetic diversity and the evolutionary history of this biome. Here we estimated the phylogenetic and populational history of the widespread Mouse-colored Tapaculo (Scytalopus speluncae) complex across the Brazilian AF, using data from two mitochondrial genes and 12 microsatellite loci. Both markers uncovered several cryptic, mostly allopatric and well-supported lineages that may represent distinct species-level taxa. We investigated whether diversification in S. speluncae is compatible with the Carnaval-Moritz model of Pleistocene refugia. We found that northern lineages have high levels of genetic diversity, agreeing with predictions of more stable forest refugia in these areas. In contrast, southern lineages have lower levels of mtDNA diversity with a signature of population expansion that occurred earlier (∼0.2Mya) than the last glacial maximum. This result suggests that the AF may be stable enough to maintain endemic taxa through glacial cycles. Moreover, we propose that the "mid-Pleistocene climate transition" between 1.2 and 0.7million years ago, from a warmer to a colder climate, may have played an important but mostly overlooked role in the evolution of AF montane taxa.

Stereological assessment of sexual dimorphism in the rat liver reveals differences in hepatocytes and Kupffer cells but not hepatic stellate cells.

There is long-standing evidence that male and female rat livers differ in enzyme activity. More recently, differences in gene expression profiling have also been found to exist; however, it is still unclear whether there is morphological expression of male/female differences in the normal liver. Such differences could help to explain features seen at the pathological level, such as the greater regenerative potential generally attributed to the female liver. In this paper, hepatocytes (HEP), Kupffer cells (KC) and hepatic stellate cells (HSC) of male and female rats were examined to investigate hypothesised differences in number, volume and spatial co-localisation of these cell types. Immunohistochemistry and design-based stereology were used to estimate total numbers, numbers per gram and mean cell volumes. The position of HSC within lobules (periportal vs. centrilobular) and their spatial proximity to KC was also assessed. In addition, flow cytometry was used to investigate the liver ploidy. In the case of HEP and KC, differences in the measured cell parameters were observed between male and female specimens; however, no such differences were detected for HSC. Female samples contained a higher number of HEP per gram, with more binucleate cells. The HEP nuclei were smaller in females, which was coincident with more abundant diploid particles in these animals. The female liver also had a greater number of KC per gram, with a lower percentage of KC in the vicinity of HSC compared with males. In this study, we document hitherto unknown morphological sexual dimorphism in the rat liver, namely in HEP and KC. These differences may account for the higher regenerative potential of the female liver and lend weight to the argument for considering the rat liver as a sexually dimorphic organ.

Genetic damage in patients moving from hemodialysis to online hemodiafiltration.

End-stage renal failure patients exhibit a high incidence of genetic damage and genomic instability. Part of this genetic damage is assumed to be caused by the hemodialysis (HD) procedure. To reduce these effects, different alternative HD procedures have been proposed, such as the use of high efficiency convective therapies to improve the reactive oxygen species/antioxidant ratio. To determine the efficiency of online hemodiafiltration (HDF) technique on the levels of DNA damage, we have measured the frequency of micronucleus in peripheral blood lymphocytes of 33 individuals moving from low-flux HD to post-dilution online HDF. In addition to basal levels of genetic damage, potential changes in radiosensitivity were measured as indicators of genomic instability. Plasma antioxidant capacity was also determined. Second samples were obtained after 6 months on the HDF protocol. Results indicate that moving to online HDF therapy produce a significant reduction of the basal levels of genetic damage, but does not affect the genomic instability status. In addition, a greater increase in plasma antioxidant capacity was observed. In spite of the lack of correlation between these parameters, our results confirm the usefulness of the online HDF technique as a way to reduce DNA damage in HD patients.

Methodological and Conceptual Limitations in Exercise Addiction Research.

The aim of this brief analytical review is to highlight and disentangle research dilemmas in the field of exercise addiction. Research examining exercise addiction is primarily based on self-reports, obtained by questionnaires (incorporating psychometrically validated instruments), and interviews, which provide a range of risk scores rather than diagnosis. Survey methodology indicates that the prevalence of risk for exercise addiction is approximately 3 percent among the exercising population. Several studies have reported a substantially greater prevalence of risk for exercise addiction in elite athletes compared to those who exercise for leisure. However, elite athletes may assign a different interpretation to the assessment tools than leisure exercisers. The present paper examines the: 1) discrepancies in the classification of exercise addiction; 2) inconsistent reporting of exercise addiction prevalence; and 3) varied interpretation of exercise addiction diagnostic tools. It is concluded that there is the need for consistent terminology, to follow-up results derived from exercise addiction instruments with interviews, and to follow a theory-driven rationale in this area of research.

Can caffeine improve your performance? Psychophysiological effects - A systematic review.

Introduction: Caffeine is a widely used ergogenic aid in society, which has made it a topic of interest due to its various benefits at cognitive, physiological, and sports levels, among others. This review aims to investigate the potential benefits of caffeine supplementation in psychophysiological performance through a structured search in the SportsDiscus/Scopus/MEDLINE and Web of Science databases (October 2022). This review followed the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guideline, and the inclusion criteria were defined based on the PICOS model. Double-blind, randomized/semi-randomized crossover articles comparing caffeine intake with an identical placebo condition were included. Filters by age or gender of the participants were not applied. The initial search gave a result of 201 articles, which after eliminating duplicates and applying the inclusion and exclusion criteria, the final sample for this review was 8 studies. The review concluded that 3 (37.5 %) found favorable ergogenic effects, 4 (50 %) found partial effects, and 1 (12.5 %) found no effects of caffeine supplementation on variables related to psychophysiological performance. In general, both partial and negative results could be linked to insufficient doses to produce any change, likewise, habitual caffeine consumption is also a variable that could be attenuating its potential ergogenic effect. In conclusion, moderate doses of caffeine 3-6 mg/kg seem to be an effective strategy to improve the psychophysiological response in various contexts without generating detrimental effects on performance, as long as the intervention designs consider the variables that could condition its effect.

Introducción: La cafeína es una ayuda ergogénica de amplio uso en la sociedad, lo que la ha convertido en un tema de interés por sus diversos beneficios a nivel cognitivo, fisiológico y deportivo, entre otros. Esta revisión tiene como objetivo investigar los beneficios potenciales de la suplementación de cafeína sobre el rendimiento psicofisiológico a través de una búsqueda estructurada en las bases de datos SportsDiscus/Scopus/MEDLINE y Web of Science (octubre de 2022). Esta revisión siguió la guía Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) y los criterios de inclusión se definieron en función del modelo PICOS. Se incluyeron artículos doble ciego, cruzados y aleatorizados/semialeatorizados en donde se comparó la ingesta de cafeína con una condición idéntica de placebo. No se aplicaron filtros por edad ni sexo de los participantes. La búsqueda inicial dio un resultado de 201 artículos, los cuales, después de eliminar los duplicados y aplicar los criterios de inclusión y exclusión, dieron una muestra final para esta revisión de 8 estudios. La revisión concluyo que 3 (37,5 %) encontraron efectos ergogénicos favorables, 4 (50 %) encontraron efectos parciales y 1 (12,5 %) no encontró efectos de la suplementación de cafeína sobre las variables relacionadas con el rendimiento psicofisiológico. En general, los resultados tanto parciales como negativos podrían estar ligados a dosis insuficientes para producir algún cambio; de igual forma, el consumo habitual de cafeína también es una variable que podría estar atenuando su potencial efecto ergogénico. En conclusión, dosis moderadas de cafeína de 3-6 mg/kg parecen ser una estrategia eficaz para mejorar la respuesta psicofisiológica en diversos contextos, sin generar efectos perjudiciales en el rendimiento, siempre y cuando los diseños de intervención consideren las variables que podrían condicionar su efecto.

Effects of a genetic counseling model on mothers of children with down syndrome: a Brazilian pilot study.

Down syndrome occurs in approximately 1:600 live births. Genetic counseling is indicated for these families and may be beneficial for adaptation to the challenges that accompany by this diagnosis. Although the basic counseling goals are similar, there are many models of genetic counseling practiced around the world. The aim of this article is to report the results of a pilot study that evaluated the level of satisfaction with a model of service delivery of genetic counseling practiced in Brazil, the knowledge assimilated about Down syndrome and whether this process resulted in a feeling of well-being and psychological support. Thirty mothers of under 6-month-old children with Down syndrome were interviewed after having two sessions of genetic counseling in a public healthcare service within a period of 30 days. A semi-structured questionnaire was developed by the researchers to collect identification, socioeconomic and demographic data and to assess the client's satisfaction with the model of genetic counseling. Data were collected using both open and closed questions. The reported level of satisfaction was high. The knowledge assimilated about Down syndrome after only two sessions was considered technically vague by raters in 44 % of cases. Most mothers (96.7 %) reported that genetic counseling was beneficial and provided psychological support. The model was considered satisfactory, but further research is needed to identify ways to improve knowledge retention by this population. These results highlight the utility of referring families for genetic counseling when there is a suspicion of a diagnosis of Down syndrome.

An immersive simulation strategy to teach cytology sample collection methods and basic diagnosis skills: A two academic center study.

BACKGROUND: Collecting cytology samples and making simple diagnoses are skills taught in veterinary universities, mostly in a passive way. Simulators enhance practical skills learning, increasing student engagement through immersive activities. These strategies have not been carefully assessed in veterinary cytology. OBJECTIVES: This study aimed to describe a simulator developed for training cytology sample collection methods and assess the utility of an immersive simulation strategy to learn and practice the collection of cytology samples. METHODS: A flipped classroom with a station design was followed. Students first watched video tutorials on sampling collection, listened to brief cases' clinical histories, and moved to immersive simulator stations. There, they practiced imprints, swabs, and fine-needle aspiration (FNA). Microscopic observation of the material was available through augmented reality tools. Students were evaluated by Objective Structured Clinical Examination (OSCE) tools on their ability to perform FNA on stuffed animal models. Students answered two questionnaires addressing their learning outcomes. RESULTS: Second- and third-year students from two centers (n = 129) practiced cytologic collection methods in simulators which significantly changed their willingness to perform FNA in live animals after the class activities. OSCE pass rates over 90% were obtained for most steps of FNA, and students rated the activity as essential/very relevant for learning. CONCLUSIONS: Immersive simulation strategies were effective at increasing student comfort with cytologic sampling techniques. This approach should be included in the veterinary curriculum as it can increase the quality of cytology samples and could potentially improve the cytologic diagnosis of a submitted sample.

Case series: Four fatal rabbit hemorrhagic disease virus infections in urban pet rabbits.

Four pet rabbits (Oryctolagus cuniculus cuniculus) diagnosed with a fatal infection by rabbit hemorrhagic disease virus (RHDV GI.2) were identified in the same week and further investigated. All animals lived in an urban environment (Lisbon, Portugal), were between 8 months and 2 years old and none had been vaccinated against RHDV2 (GI.2). Three animals arrived at the clinic and died shortly afterward and it was only possible to collect material for RT-qPCR (RHDV) test. These rabbits tested positive for RHDV2, with high viral loads. In the fourth case, additional clinical and post-mortem gross and histological evaluations were performed. This 8 month old intact female indoor pet rabbit was presented with apathy, tachypnea and tachycardia. Radiographic projections revealed no clinical revealed no clinical abnormalities. Serum biochemistry revealed a significant increase in AST and ALT with a small hypoglycemia. Abdominal ultrasound revealed an acute hepatitis. Despite hospitalization support, after 30 h of admission, the rabbit lost consciousness and developed anorexia and pyrexia in the last minutes before death. Post-mortem analysis and molecular testing by RT-qPCR, confirmed the diagnosis of RHDV2 (GI.2) infection also with high viral load. In conclusion, this paper reports a case series that demonstrates the severe infectious ability and the high mortality associated with RHDV even in rabbits from urban environments. Furthermore, it highlights the importance of always considering rabbit hemorrhagic disease (RHD) as a differential diagnosis in pet rabbits with non-specific clinical signs, and should warn veterinarians that pet rabbits living indoors can also be infected with a fatal outcome.

The Use of Simulation Models and Student-Owned Animals for Teaching Clinical Examination Procedures in Veterinary Medicine.

Clinical examination procedures (CEPs) are cornerstone clinical skills for veterinary practitioners, being taught in all veterinary faculties. CEPs include innocuous procedures that are well tolerated by animals as well as more distressful and less tolerated ones. In a classical approach, institutional animals are used to teach and practice CEPs. Two hundred and thirty-one undergraduate students from four consecutive years were assigned to two groups that used institutional animals only (AO) or a combination of students' owned animals and simulation models (model-animal, MA) to teach and practice CEPs. This latter comprised stuffed teddy dogs, eye and ear models made of molding silicone, as well as skin models. The learning outcome of each system was compared through questionnaires (throughout classes and at the end of course), grades, and pass rates in objectively structured clinical examinations. Most veterinary students had their own animals, being easy to have a dog per group of two students in class. All the students' owned animals adapted well to this environment. The interest in the practical activities with the simulation models was comparable to that exhibited in the classical AO system. Students reported to learn more with the MA system than with the AO, while the interest on the subjects and the relevance were appraised similarly in both systems. No differences existed in the final grades and pass rates. The MA system was effective for learning CEPs. Beyond animal welfare advantages, this system increased the out-of-school training and had financial saving benefits, being a valuable option for the teaching and training of CEPs.

Evaluation of a Manual Cytocentrifuge versus the Standard Automated Cytocentrifuge in the Analysis of Canine Cerebrospinal Fluid: A Case Series of 55 Dogs.

Cytospins are important for evaluating fluids with very low cellularity such as cerebrospinal fluid (CSF). The aim of this study was to compare the CSF cytospin preparations obtained from automated and manual cytocentrifugation methods. A prospective case series was performed to analyze canine CSF samples using both centrifugation methods. The cytospins were processed within 30-60 min and prepared simultaneously in a conventional automated cytocentrifuge and in an in-house manual cytocentrifuge, using a fixed volume of CSF fluid. The cellularity, differential cell count and the proportion of cell artifacts (pseudopods and vacuolization) were blindly assessed in the cytospin preparations obtained using the two methods. The agreement and correlation between both methods were analyzed. There were 55 dogs enrolled (48 prospectively and 7 retrospectively) in the study. 38 dogs had normal total nucleated cell counts, while 17 had pleocytosis. Automated and manual cytocentrifugation had similar cell yields, and no significant differences in differential cell counts or the presence of artifacts existed between both methods. In cases with pleocytosis, the cytologic diagnosis obtained using each method was similar. Manual cytocentrifugation of CSF is a reliable and economic method designed for routine clinical practice. Its use reduces the specimen deterioration related to processing and analysis delays when samples are transported to external laboratories for evaluation.

Detection of Lymphoid Markers (CD3 and PAX5) for Immunophenotyping in Dogs and Cats: Comparison of Stained Cytology Slides and Matched Cell Blocks.

Immunolabeling on Romanowsky-stained cytology (RSC) slides can be used, although there is limited evidence of its suitability for phenotyping canine and feline lymphomas. A comparison with matched cell blocks (CB) is missing. Immunolabeling on RSC and CB was compared for lymphoid markers (CD3 and PAX5) in 53 lymphomas and 4 chylous effusions from dogs and cats. The influence of pre-analytical variables (species, time of archive, type of specimens and coverslipping) and the interobserver agreement among the 2 observers was assessed. Fewer CD3+ lymphocytes were identified in RSC, while the PAX5 positivity by RSC and CB had a substantial agreement. Immunodetection of CD3 and the diagnosis of a T-cell population on RSC was more difficult. Lower intensity and higher background were noted in RSC. Immunophenotyping was inconclusive in 54% RSC and 19% CB. The interobserver reproducibility of immunophenotyping on CB was substantial, being higher than in RSC. The immunolabeling performance on the RSC of effusion and feline samples was unsatisfactory. The detection of lymphoid markers, especially membranous antigens in retrospective RSC, is affected by the pre-analytical variables: species, time of the archive, and type of specimens. CB are a more consistent type of sample for immunophenotyping purposes.

Influence of Exogenous Factors Related to Nutritional and Hydration Strategies and Environmental Conditions on Fatigue in Endurance Sports: A Systematic Review with Meta-Analysis.

The aim of this systematic review with meta-analysis was to examine the influence of exogenous factors related to nutritional and hydration strategies and environmental conditions, as modulators of fatigue, including factors associated with performance fatigability and perceived fatigability, in endurance tests lasting 45 min to 3 h. A search was carried out using four databases: PubMed, Web of Science, SPORTDiscus, and EBSCO. A total of 5103 articles were screened, with 34 included in the meta-analysis. The review was registered with PROSPERO (CRD42022327203) and adhered to the PRISMA guidelines. The study quality was evaluated according to the PEDro score and assessed using Rosenthal's fail-safe N. Carbohydrate (CHO) intake increased the time to exhaustion (p < 0.001) and decreased the heart rate (HR) during the test (p = 0.018). Carbohydrate with protein intake (CHO + PROT) increased lactate during the test (p = 0.039). With respect to hydration, dehydrated individuals showed a higher rate of perceived exertion (RPE) (p = 0.016) and had a higher body mass loss (p = 0.018). In hot conditions, athletes showed significant increases in RPE (p < 0.001), HR (p < 0.001), and skin temperature (p = 0.002), and a decrease in the temperature gradient (p < 0.001) after the test. No differences were found when athletes were subjected to altitude or cold conditions. In conclusion, the results revealed that exogenous factors, such as nutritional and hydration strategies, as well as environmental conditions, affected fatigue in endurance sports, including factors associated with performance fatigability and perceived fatigability.