Cryopreservation of canine ovarian tissue by slow freezing and vitrification: Evaluation of follicular morphology and apoptosis rate.

In this study, we aimed to evaluate the efficacy of cryopreserving canine ovarian tissue using vitrification and slow freezing methods while investigating potential differences in cryotolerance based on follicular type and cryopreservation technique. Twenty-eight ovaries were collected from 14 anoestrus bitches of various breeds, aged between 2 and 5 years, and undergoing elective ovariohysterectomy. The ovaries were sectioned into small fragments and randomly assigned to three groups: vitrification, slow freezing, and a control group (fresh tissue). Vitrification was performed using cryotubes containing DAP 213 solution (2M DMSO, 1M acetamide, 3M propylene glycol) in two stages, while slow freezing involved cryotubes with 1.5M DMSO solution inserted into a programmable machine. The effects of cryopreservation were evaluated by histology and immunohistochemistry (cleaved caspase-3), to determine the percentage of cells undergoing apoptosis. Histological examination revealed that the slow freezing group exhibited a significantly higher percentage of intact follicles (45.75 %) compared to those subjected to vitrification (38.17 %; P = 0.01). Immunohistochemical evaluation further indicated that 84.21 % of the follicles in the slow freezing group did not express caspase-3, suggesting the absence of apoptosis. Conversely, vitrified samples exhibited significantly more apoptotic cells compared to other groups (P < 0.001). Furthermore, early antral follicles displayed a higher susceptibility to degeneration regardless of the cryopreservation method employed. Nevertheless, when comparing the cryopreserved groups, early antral follicles showed greater degeneration in slow freezing group, while preantral follicles were the most affected in the vitrification group. In conclusion, slow freezing demonstrated superior preservation of viable follicles compared to vitrification and emerged as the preferred technique for cryopreserving canine ovarian tissue. These findings contribute valuable insights into optimizing cryopreservation methods for canine ovarian tissue, potentially benefiting reproductive technologies and fertility preservation in canines.

Serum Extracellular Vesicles Cargo Approach in Bitches with Mammary Tumors.

This study investigated serum extracellular vesicles (EVs) in bitches with mammary neoplasms, in order to understand their size, shape, and concentration, as well as their association with tumor malignancy. Thirty bitches were categorized into control (n = 10), mammary tumor grades I and II (GI, n = 13), and grade III (GII, n = 7). Serum was separated from blood collected during mastectomy, and EVs were isolated using size exclusion chromatography. The analysis revealed no significant differences in EV concentrations among groups, with similar concentrations for control, GI, and GII. Ninety-one proteins were identified in EV-enriched samples, with six showing varied abundance across groups. Notably, keratin 18 was highly abundant in GI, while sushi domain-containing protein, EvC ciliary subunit 2, and the joining chain of multimeric IgM and IgA were increased in GII. Additionally, protocadherin 17 and albumin were upregulated in both GI and GII. ROC curves identified potential biomarkers for differentiating tumor grades. Enrichment pathway analysis revealed AFP gene upregulation in the GI. Mass spectrometry proteomics data were deposited in Mendeley Data. The study provides valuable insights into serum EV characterization in bitches, suggesting keratin 18 and protocadherin 17 as potential biomarkers for canine mammary neoplasia, with implications for future diagnostic and therapeutic strategies.

Exploring Canine Mammary Cancer through Liquid Biopsy: Proteomic Profiling of Small Extracellular Vesicles.

(Background). Canine mammary tumors (CMTs) have emerged as an important model for understanding pathophysiological aspects of human disease. Liquid biopsy (LB), which relies on blood-borne biomarkers and offers minimal invasiveness, holds promise for reflecting the disease status of patients. Small extracellular vesicles (SEVs) and their protein cargo have recently gained attention as potential tools for disease screening and monitoring. (Objectives). This study aimed to isolate SEVs from canine patients and analyze their proteomic profile to assess their diagnostic and prognostic potential. (Methods). Plasma samples were collected from female dogs grouped into CMT (malignant and benign), healthy controls, relapse, and remission groups. SEVs were isolated and characterized using ultracentrifugation (UC), nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Proteomic analysis of circulating SEVs was conducted using liquid chromatography-mass spectrometry (LC-MS). (Results). While no significant differences were observed in the concentration and size of exosomes among the studied groups, proteomic profiling revealed important variations. Mass spectrometry identified exclusive proteins that could serve as potential biomarkers for mammary cancer. These included Inter-alpha-trypsin inhibitor heavy chain (ITIH2 and ITI4), phosphopyruvate hydratase or alpha enolase (ENO1), eukaryotic translation elongation factor 2 (eEF2), actin (ACTB), transthyretin (TTR), beta-2-glycoprotein 1 (APOH) and gelsolin (GSN) found in female dogs with malignant tumors. Additionally, vitamin D-binding protein (VDBP), also known as group-specific component (GC), was identified as a protein present during remission. (Conclusions). The results underscore the potential of proteins found in SEVs as valuable biomarkers in CMTs. Despite the lack of differences in vesicle concentration and size between the groups, the analysis of protein content revealed promising markers with potential applications in CMT diagnosis and monitoring. These findings suggest a novel approach in the development of more precise and effective diagnostic tools for this challenging clinical condition.

Platelet-Rich Plasma Proteome of Mares Susceptible to Persistent-Breeding-Induced Endometritis Differs from Resistant Mares.

Persistent-breeding-induced endometritis (PBIE) is the leading cause of subfertility and poor reproductive efficiency in mares. Platelet-rich plasma (PRP) treatment has been shown to mitigate PBIE, reduce uterine infections, and improve fertility in mares. However, the proteome of PRP in mares, particularly those susceptible to PBIE, remains unknown. This study aimed to fill this knowledge gap by comparing the most abundant proteins present in PRP prepared from mares with histories of being susceptible or resistant to PBIE. The study involved twelve light-breed mares: seven susceptible and five resistant to PBIE. A complete blood count and physical examination were performed on each mare before blood drawing to ensure good health. The PRP was prepared following collection in a blood transfusion bag and double centrifugation. Platelet counts in the PRP were compared across the groups. The PRP was cryopreserved in liquid nitrogen until proteomics could be completed. Physical parameters and complete blood cell counts were within normal ranges. The platelet counts for resistant (561 ± 152 × 103) and susceptible mares (768 ± 395 × 103) differed (p < 0.05). One hundred and five proteins were detected in all mares, and four proteins were more abundant in resistant mares (p < 0.05). The proteins were apolipoprotein C-II, serpin family G member 1, protection of telomeres protein 1, and non-specific serine/threonine protein kinase. All these proteins are linked to the immune response. These results suggest that PRP prepared from mares resistant to PBIE may be more beneficial in mitigating PBIE in mares, offering a promising avenue for improving equine reproductive health. However, this remains to be determined with in vivo studies.

Neonatal hypoglycemia in dogs-pathophysiology, risk factors, diagnosis and treatment.

Hypoglycemia is the most common metabolic alteration in the clinical routine of newborn dogs, acting as a predictor of mortality in these patients. The neonatal dog shows hepatic insufficiency and homeostatic mechanisms not yet fully developed, with limited glycogen reserves and limited capacity to perform glycogenolysis and gluconeogenesis. These physiological particularities make newborn dogs particularly susceptible to hypoglycemia when of fasting, even for short periods. Several maternal and neonatal factors may be related to a higher risk of developing hypoglycemia in neonates. This paper reviews glycemic homeostasis, the pathophysiology of neonatal hypoglycemia, the main causes involved and the diagnostic and therapeutic approaches to this condition.

The interaction between seminal plasma, sperm, and endometrium in inter- and intra-species breeding in equids.

In brief: In some instances, extra-species breeding in equids is more successful than intraspecies breeding; however, little is known about the immunomodulatory effect of donkey semen and seminal plasma on the mare's endometrium. This study compared the mare uterine inflammatory response during extra- and intraspecies breeding. Abstract: Anecdotal experience suggests horse mares have less post-breeding inflammation and better fertility when bred with donkeys. This study aimed to compare the post-breeding inflammatory response of mares exposed to donkey and horse semen and seminal plasma and evaluate the proteome and metabolome of donkey and horse sperm and seminal plasma. Uterine edema, intrauterine fluid accumulation, polymorphonuclear neutrophils on cytology, and concentrations of progesterone, and pro- and anti-inflammatory cytokines (IL1A, IL1B, IL4, IL6, CXCL8, IL10) were assessed pre- and post infusion of semen and seminal plasma (donkey and horse). The metabolome and proteome were analyzed by LC-MS/MS. Mare cycles bred with horse semen had a greater progesterone concentration than those bred with donkey semen at 8 days post ovulation (P = 0.046). At 6 h post infusion, the inflammatory response due to the donkey semen tended to be lower (P = 0.074). Donkey seminal plasma had anti-inflammatory properties compared to horse semen and seminal plasma, as determined by fewer neutrophils on uterine cytology (P < 0.05). Horse semen resulted in greater concentrations of IL6 and lesser concentrations of IL1B (P < 0.05). PGE1, PGE3, and lactoferrin concentrations were significantly more abundant in donkey sperm and seminal plasma. Prostaglandins play an important role in immunomodulation and might contribute to the response triggered in interspecies breeding. In conclusion, breeding horse mares with donkey semen induces similar post-breeding endometritis as observed with horse semen. Donkey seminal plasma results in a lower post-infusion inflammatory response compared to other combinations in the immediate post-breeding.

Symmetric dimethylarginine correlates with the urea, creatinine, potassium, and clinical scores in feline urethral obstructions.

BACKGROUND: A urethral obstruction (UO) is an emergency commonly observed in male cats, which can result in significant clinical and laboratory alterations, leading to complications and death. OBJECTIVES: This study aimed to correlate symmetric dimethylarginine (SDMA) with the urea, creatinine, potassium, and bicarbonate levels in cats with UO. In addition, the correlation between clinical score and time of obstruction was evaluated. METHODS: Thirty male cats were selected and allocated into a control group (CG, n = 13) and an obstruction group (OG, n = 17). The laboratory analyses were conducted before treatment (M0) and at different times after treatment (12 h [M12], 24 h [M24], and 48 h [M48]). Correlations were established between SDMA and creatinine, urea, bicarbonate, potassium, time of obstruction, and the clinical score. RESULTS: A strong correlation (r > 0.6) was observed between SDMA and creatinine, urea, and potassium in the OG. Furthermore, there was substantial agreement (kappa value) between SDMA and creatinine at M24. A higher clinical score was associated with a longer time of obstruction. In the OG, at M48, the SDMA and creatinine levels were 50% and 41.2% higher, respectively. CONCLUSIONS: A correlation was observed between SDMA and creatinine in obstructed cats, and significant agreement between these values was observed 24 h after the unblocking treatment. A correlation among SDMA, urea, and potassium was observed. Approximately 9% more cats continued to have elevated SDMA levels after 48 h of treatment compared to creatinine. This suggests a slightly lower sensitivity of the latter biomarker but does not exclude the possibility of congruent and normalized values after a longer evaluation period.

High depressive symptomatology reduces emotional reactions to pictures of social interaction.

Individuals with severe depressive symptoms present diminished facial expressions compared to healthy individuals. This reduced facial expression, which occurs in most depressive patients could impair social relationships. The current study sought to investigate whether pictures with social interaction cues could elicit different modulations of facial expressions and mood states in individuals with depressive symptoms compared to healthy individuals. A total of 85 individuals were divided into depressive and non-depressive groups based on their beck depression inventory scores. Participants viewed pictures containing neutral (objects), affiliative (people interacting socially), and control (people not interacting) scenes. Electromyographic signals were collected during the entire period of visualization of the blocks, and emotional questionnaires were evaluated after each block to assess sociability and altruism (prosocial states). In non-depressed individuals, affiliative pictures increased the activity of the zygomatic muscle compared to both neutral and control pictures and reduced fear of rejection compared to neutral pictures. During the visualization of the affiliative block, zygomatic major muscle activation was higher and fear of rejection was lower in the non-depressive individuals than in the depressive. These effects reflected the low expressions of smiling and sociability to affiliative pictures in depressive individuals. These findings highlight the importance of smiling and prosocial states in social interactions, especially in these individuals.

Uterine secretome: What do the proteins say about maternal-fetal communication in buffaloes?

The aim was to compare the UF proteomics of pregnant and non-pregnant buffalo during early pregnancy. Forty-four females were submitted to hormonal estrus synchronization and randomly divided into two groups: pregnant (n = 30) and non-pregnant (n = 14). The pregnant group was artificially inseminated and divided into a further two groups: P12 (n = 15) and P18 (n = 15). Conceptus and uterine fluid samples were collected during slaughter at, respectively, 12 and 18 days after insemination. Of all the inseminated females, only eight animals in each group were pregnant, which reduced the sample of the groups to P12 (n = 8) and P18 (n = 8). The non-pregnant group was also re-divided into two groups at the end of synchronization: NP12 (n = 7) and NP18 (n = 7). The UF samples were processed for proteomic analysis. The results were submitted to multivariate and univariate analysis. A total of 1068 proteins were found in the uterine fluid in both groups. Our results describe proteins involved in the conceptus elongation and maternal recognition of pregnancy, and their action was associated with cell growth, endometrial remodeling, and modulation of immune and antioxidant protection, mechanisms necessary for embryonic maintenance in the uterine environment. SIGNIFICANCE: Uterine fluid is a substance synthesized and secreted by the endometrium that plays essential roles during pregnancy in ruminants, contributing significantly to embryonic development. Understanding the functions that the proteins present in the UF perform during early pregnancy, a period marked by embryonic implantation, and maternal recognition of pregnancy is of fundamental importance to understanding the mechanisms necessary for the maintenance of pregnancy. The present study characterized and compared the UF proteome at the beginning of pregnancy in pregnant and non-pregnant buffaloes to correlate the functions of the proteins and the stage of development of the conceptus and unravel their processes in maternal recognition of pregnancy. The proteins found were involved in cell growth and endometrial remodeling, in addition to acting in the immunological protection of the conceptus and performing antioxidant actions necessary for establishing a pregnancy.

Proteomics approach reveals urinary markers for early pregnancy diagnosis in buffaloes.

This study aimed to compare urine proteomics from non- and pregnant buffaloes in order to identify potential biomarkers of early pregnancy. Forty-four females underwent hormonal ovulation synchronization and were randomly divided into two experimental groups: inseminated (n = 30) and non-inseminated (n = 14). The pregnant females were further divided into two groups: pregnant at Day 12 (P12; n = 8) and at Day 18 (P18; n = 8) post-ovulation. The non-pregnant group was also subdivided into two groups: non-pregnant at Day 12 (NP12; n = 7) and at Day 18 (NP18; n = 7). Urine was collected from all females on Days 12 or 18. The samples were processed for proteomics. A total of 798 proteins were reported in the urine considering all groups. The differential proteins play essential roles during pregnancy, acting in cellular transport and metabolism, endometrial remodeling, embryonic protection, and degradation of defective proteins. We suggest that some proteins from our study can be considered biomarkers for early pregnancy diagnosis, since they were increased in pregnant buffaloes. SIGNIFICANCE: Macromolecules have been studied for early pregnancy diagnosis, aiming to increase reproductive efficiency in cattle and buffaloes. Direct methods such as rectal palpation and ultrasonography have been considered late. Thus, this study aimed to compare urine proteomics from non- and pregnant buffaloes to identify potential biomarkers of early pregnancy. The differential proteins found in our study play essential roles during pregnancy, acting in cellular transport and metabolism, endometrial remodeling, embryonic protection, and degradation of defective proteins. We suggest that these proteins can be considered possible biomarkers for early pregnancy diagnosis since they were increased in the pregnant buffaloes.

"Como será minha vida com sífilis?" Desafios do diagnóstico em homens à luz de Leininger / "How will my life with syphilis be?" Diagnosis challenges in men in the light of Leininger / ¿Cómo será mi vida con sífilis? Desafíos diagnósticos en los hombres a la luz de Leininger

Objetivo: compreender os fatores que interferem no diagnóstico da sífilis em homens à luz da Teoria da Diversidade e Universalidade do Cuidado Cultural. Método: estudo qualitativo, aprovado por Comitê de Ética em Pesquisa, realizado em Hospital Universitário no Rio de Janeiro, entre setembro de 2017 e março de 2018. Foram entrevistados 32 homens diagnosticados com sífilis adquirida, e a análise temática foi utilizada para o tratamento dos dados. Resultados: a maioria dos homens tinha história de contágio por outra Infecção Sexualmente Transmissível e foi diagnosticada no ambulatório de imunologia. O momento diagnóstico é encarado com surpresa, tem repercussões psicossociais e é influenciado por fatores culturais e sociais. Considerações finais: alguns fatores interferem positivamente e outros negativamente no diagnóstico da sífilis na população masculina. Para detectar essa infecção nos homens, deve-se conhecer o contexto sociocultural em que estão inseridos para, assim, implementar estratégias tanto diagnósticas quanto preventivas mais eficazes(AU)

Objective: to understand the factors that interfere with the diagnosis of syphilis in men in the light of the Theory of Diversity and Universality of Cultural Care. Method: qualitative study, approved by the Ethics and Research Committee, conducted at a University Hospital in Rio de Janeiro, during september 2017 to march 2018. Thirty-two men diagnosed with acquired syphilis were interviewed, and thematic analysis was used for data treatment. Results: most men had a history of contagion by another STI and were diagnosed at the immunology outpatient clinic. The moment of diagnosis is faced with surprise, has psychosocial repercussions and is influenced by cultural and social factors. Final considerations: some factors interfere positively and others negatively in the diagnosis of syphilis in the male population. To detect this infection in men, it is necessary to know the sociocultural context in which they live, in order to implement more effective diagnostic and preventive strategies(AU)

Objetivo: comprender los factores que interfieren en el diagnóstico de la sífilis en los hombres a la luz de la Teoría de la diversidad y universalidad del cuidado cultural. Método: estudio cualitativo, aprobado por el Comité de Ética en Investigación, realizado en un Hospital Universitario de Río de Janeiro, de septiembre de 2017 a marzo de 2018. Se entrevistó a 32 hombres diagnosticados de sífilis adquirida y se utilizó el análisis temático para el tratamiento de los datos. Resultados: la mayoría de los hombres tenía antecedentes de contagio por otra Infección Sexualmente Transmisible y el diagnóstico se hizo en el ambulatorio de inmunología. El momento del diagnóstico se afronta con sorpresa, tiene repercusiones psicosociales y está influenciado por factores culturales y sociales. Consideraciones finales: algunos factores interfieren positivamente y otros negativamente en el diagnóstico de la sífilis en la población masculina. Para detectar esta infección en los hombres, se debe conocer el contexto sociocultural en el que se insertan, para entonces poner en marcha estrategias de diagnóstico y de prevención más eficaces(AU)

Hypoglycaemia management with a hypercaloric supplementation in neonatal puppies delivered by caesarean section.

There is a high perinatal mortality rate in dogs, estimated at 20%, and one of the leading causes of this rate is hypoglycaemia. Therefore, we aimed to evaluate the efficacy of a hypercaloric supplement containing vitamins and amino acids in newborn puppies presenting hypoglycaemia at birth. Ninety-nine pups were divided into four groups: normoglycaemic caesarean section (NORMOCS), hypoglycaemic caesarean section supplemented with the hypercaloric (SUPLCS), hypoglycaemic caesarean section supplemented with glucose (GLICCS) and eutocic delivery (EUT). We evaluated the following parameters glycaemia, Apgar score, neurological reflexes and rectal temperature of neonates at the following moments 5 min (M5), 30 min (M30) and 60 min (M60) after birth. Brachycephalic dogs were 73.3% (22/30) of caesarean sections (c-sections). The puppy's average glycaemia represented about 90% of the maternal glycaemia, while 15.1% (14/99) of the neonates had hypoglycaemia (<90 mg/dL) at M0 and 46.5% (44/99) at M60. Only four neonates had glycaemia below 40 mg/dL at M30 but without showing any clinical signs. The puppy's fasting while waiting for the intraoperative period and the dam's anaesthetic recovery was considered risk factors for hypoglycaemia. There was no difference in mean blood glucose levels or vitality parameters among puppies from the SUPLCS and GLICCS. In conclusion, the hypercaloric supplement can be used as a replacement for glucose in hypoglycaemic puppies and it can also bring nutritional benefits for the puppy. The prepartum glycaemia of the dam is an important parameter to be measured, and the appropriate management of it reduces the chances of the puppies being born with hypoglycaemia.

Ecological aspects of Pintomyia fischeri and Migonemyia migonei in municipalities with Canine Visceral Leishmaniasis, State of São Paulo, Brazil.

The objective was to study the composition of sand fly species, the seasonal activity of the dominant species and correlation with average rainfall, sex association of the dominant species with domicile habitats and analyze different landscape scenarios for the two study sites in the municipalities of Itapevi and Mogi of the Crosses. Sandflies were captured with CDC(s) traps installed in three households at each study site for a period of 24 hours, biweekly for two years. A total of 2970 specimens were collected. The dominant species were P.fischeri and M.migonei. A statistically significant difference was registered between males and females of the two species in relation to the intra and peridomicile. The seasonal activity of both species showed a weak significant positive correlation with rainfall. The species P.fischeri and M.migonei may be potential vectors of CVL in the studied sites.

Impact of a quality improvement programme on the preparation and administration of medications via a nasoenteral feeding tube: 2014-2019 intervention study.

AIMS: Evaluate the impact of a quality improvement programme on the reduction of feeding tube obstruction frequencies, analyse the predictive variables of this safety incident, and estimate the economic costs related to the quality improvement programme during the period from 2014 to 2019. METHODS: Plan-Do-Study-Act cycles were performed to test the changes in drug preparation and administration processes via a nasoenteral feeding tube and to evaluate the outcome, process and balance measures. Statistical control charts were elaborated, and the bottom-up direct costing methodology was used to estimate the costs of the improvement programme. The impact of the programme on the monitoring measures was evaluated using logistic regression analysis. INTERVENTIONS: The following changes were tested in the hospital participating in the study: acquisition of the Easy Crush equipment for tablet crushing, use of appropriate packaging to crush hard tablets, standardise procedures for scheduling administration times and/or substitution of the pharmaceutical form, educational activities for the nursing team and elaboration and availability of infographics for the nursing team, patients and/or family/caregivers. RESULTS: There was a significant improvement in the frequency of tube obstructions, from 41.1% in 2014 to 57.9% in 2015-2017 and 9.6% in 2018-2019 (p=0.0010). After the execution of the improvement programme, it was estimated that the cost of dose preparation was reduced from R$1067.50 in 2014 to R$719.80 in 2015-2017 and R$433.10 in 2015-2019. CONCLUSION: By re-establishing the processes of drug preparation and administration via a nasoenteral feeding tube, through the acquisition of appropriate equipment for crushing hard tablets, together with educational activities for the nursing team, we could observe a reduction in tube obstructions and the cost of processes.

Influence of culture conditions on the secretome of mesenchymal stem cells derived from feline adipose tissue: Proteomics approach.

This study aimed to describe the secretome of mesenchymal stem cells derived from feline adipose tissue (AD-MSCs) and compare the effects of different culture conditions on AD-MSC proteomics using a shotgun approach. Adipose tissue was collected from 5 female cats and prepared to culture. Conditioned media was collected at third passage, in which the cells were cultured under 4 conditions, normoxia with fetal bovine serum (N + FBS), hypoxia with FBS (H + FBS), normoxia without FBS (N - FBS), and hypoxia without FBS (H - FBS). Then, the secretome was concentrated and prepared for proteomic approaches. Secretomes cultured with FBS-free medium had more than twice identified proteins in comparison with the secretomes cultured with FBS. In contrast, hypoxic conditions did not increase protein amount and affected only a small proteome fraction. Relevant proteins were related to the extracellular matrix promoting environmental modulation, influencing cell signaling pathways, and providing a suitable environment for cell proliferation and maintenance. Moreover, other proteins were also related to cell adhesion, migration and morphogenesis. Culture conditions can influence protein abundance in AD-MSC secretome, and can give also more specificity to cell and cell-free treatments for different diseases.

Pediatric allergy and immunology for patients and parents: challenges of developing website and social network during COVID-19 pandemic in Brazil.

OBJECTIVE: To describe the development of a website and the creation of a social network account about pediatric allergy/immunology with reliable information, to promote education and have a channel for patient-doctor contact. METHODS: This is a descriptive study. A survey was conducted with 93 patients (12 years and older) and caregivers of a Pediatric Allergy/Immunology outpatient clinic, to assess internet usage patterns of potential users. A webpage in Portuguese and an Instagram® account were launched in which it was created an area for patient-doctor communication in the pandemic context. RESULTS: Among 93 participants, 77% were female, 82% caregivers. Median age was 33.2 years, family income 403 dollars/month. The internet was accessed via smartphone by 81,7% of the participants; 76% reported using internet to access health information but 72% did not trust on the information from the internet, and 96% believed that an institutional site could provide meaningful information. From the website release in November 6, 2018 to January 20, 2022, it was counted 10,062 page views by 4,896 users; 55% were 18-34 years old, 70.2% female. Instagram® account gathered 882 followers. Website went through a period of instability during which access were not counted. Due to social isolation during COVID-19 pandemic, the website served as a tool for first response to help patients and doctors. CONCLUSIONS: Patients and caregivers of the Pediatric Allergy/Immunology service, consulted about digital tools, considered the information supported by a teaching/research institution timely and relevant. The website and Instagram® account have both performed well and shown good return in relation to hits, and results are continuously being evaluated. During COVID-19 pandemic, the website has been connecting patients/families and doctors.

Multiple flow cytometry analysis for assessing human sperm functional characteristics.

Multiparametric analysis by flow cytometry solves one of the major problems in sperm evaluation, the inability to test multiple attributes simultaneously in a single cell, which would increase the precision to predict fertility potential since several sperm parameters are tested. The association of fluorochromes and compounds conjugated to fluorochromes in multiparametric sperm analysis is well-established in microscopy techniques. However, these techniques are subjective and limit the assessment in small cell numbers, thereby harming analytic accuracy. Therefore, the current study aimed to present new possibilities for assessing the integrity and stability of the sperm plasma membrane, acrosome status, mitochondrial potential, and superoxide anion production in the mitochondrial matrix in only 2 cytometric assays using cytometers equipped with 2 and 3 lasers. For this, human semen samples collected by masturbation and selected by the swim-up technique were divided into 3 treatments: T0 (flash-frozen semen), T50 (flash-frozen semen + fresh semen, V: V), and T100 (fresh semen) for the validation of the multiparametric protocols by flow cytometry. For both protocols, sperm percentage with positive stain for all fluorophores differed significantly between treatments. The determination coefficients presented values close to 1, which validated objective, sensitive, rapid, and reproducible methodologies. Therefore, we concluded that the results reflect the status of analyzed structure, enabling a more accurate diagnosis of male infertility that has become an increasingly prevalent worldwide setback due to exposure to a variety of environmental toxicants.

Puerperal women's perceptions of obstetric violence during labor / Percepção de puérperas sobre a violência obstétrica durante o trabalho de parto

INTRODUCTION: When perceptions from healthcare users regarding the services offered by health institutions are taken into account, they become a tool to improve the quality of healthcare actions. OBJECTIVE: To understand the perception of puerperal women about obstetric violence during labor. METHODS: A descriptive mixed-methods study was carried out in a maternity hospital in the city of São Paulo with 31 postpartum women from August to November 2020. Quantitative data were collected in the first step, followed by the collection and analysis of qualitative data based on the theoretical framework of social representations and content analysis to decompose the interview material. RESULTS: Two categories were identified from the perspective of the obstetric violence perception, physical abuse, and abandonment during health care. From the perceptions of puerperal women concerning obstetric violence during labor, it was possible to identify that 11 women have experienced it. Of these, 7 did not initially mention violence, but when questioned, they indicated the occurrence of inappropriate or unnecessary practices that fit the obstetric violence definition, and 20 claimed not to have experienced violence. CONCLUSION: It was evidenced that women sometimes did not recognize that they experienced obstetric violence, and some of those women who did had received inadequate information. There is an urgent need to reflect on the current scenario of childbirth care, reformulation of care practices, and professional awareness of good practices and evidence-based medicine.

Point-of-care ultrasonography to predict fluid responsiveness in children: A systematic review and meta-analysis.

BACKGROUND: Point-of-care ultrasonography (POCUS) is proposed as a valuable method for hemodynamic monitoring and several ultrasound-based predictors of fluid responsiveness have been studied. The main objective of this study was to assess the accuracy of these predictors in children. METHODS: PubMed, Embase, Scopus, ClinicalTrials.gov, and Cochrane Library databases were searched for relevant publications through July 2022. Pediatric studies reporting accuracy estimates of ultrasonographic predictors of fluid responsiveness were included since they had used a standard definition of fluid responsiveness and had performed an adequate fluid challenge. RESULTS: Twenty-three studies involving 1028 fluid boluses were included, and 12 predictors were identified. A positive response to fluid infusion was observed in 59.7% of cases. The vast majority of participants were mechanically ventilated (93.4%). The respiratory variation in aortic blood flow peak velocity (∆Vpeak) was the most studied predictor, followed by the respiratory variation in inferior vena cava diameter (∆IVC). The pooled sensitivity and specificity of ∆Vpeak were 0.84 (95% CI, 0.76-0.90) and 0.82 (95% CI, 0.75-0.87), respectively, and the area under the summary receiver operating characteristic curve (AUSROC) was 0.89 (95% CI, 0.86-0.92). The ∆IVC presented a pooled sensitivity and specificity of 0.79 (95% CI, 0.62-0.90) and 0.70 (95% CI, 0.51-0.84), respectively, and an AUSROC of 0.81 (95% CI, 0.78-0.85). Significant heterogeneity in accuracy estimates across studies was observed. CONCLUSIONS: POCUS has the potential to accurately predict fluid responsiveness in children. However, only ∆Vpeak was found to be a reliable predictor. There is a lack of evidence supporting the use of POCUS to guide fluid therapy in spontaneously breathing children.