Zebrafish Larvae Microinjection and Automated Fluorescence Microscopy for Studying Klebsiella pneumoniae Infection and the Host Immune Response.

Studying host-pathogen interactions is essential for understanding infectious diseases and developing possible treatments, especially for priority pathogens with increased virulence and antibiotic resistance, such as Klebsiella pneumoniae. Over time, this subject has been approached from different perspectives, often using mammal host models and invasive endpoint measurements (e.g., sacrifice and organ extraction). However, taking advantage of technological advances, it is now possible to follow the infective process by noninvasive visualization in real time, using optically amenable surrogate hosts. In this line, this chapter describes a live-cell imaging approach to monitor the interaction of K. pneumoniae and potentially other bacterial pathogens with zebrafish larvae in vivo. This methodology is based on the microinjection of fluorescent bacteria into the otic vesicle, followed by time-lapse observation by automated fluorescence microscopy with environmental control, monitoring the dynamics of immune cell recruitment, bacterial load, and larvae survival.

Cholecystectomy and digestive cancer in Chile: Complementary results from interrupted time series and aggregated data analyses.

Gallbladder cancer (GBC) mortality in Chile is among the highest worldwide. In 2006, the Chilean government launched a programme guaranteeing access to gallbladder surgery (cholecystectomy) for patients aged 35-49 years. We evaluated the impact of this programme on digestive cancer mortality. After conducting an interrupted time series analysis of hospitalisation and mortality data from 2002 to 2018 publicly available from the Chilean Department of Health Statistics and Information, we calculated the change in the proportion of individuals without gallbladder since 10 years. We then estimated age, gender, region, and calendar-year standardised mortality ratios (SMRs) as a function of the change in the proportion of individuals without gallbladder. The cholecystectomy rate increased by 45 operations per 100,000 persons per year (95%CI 19-72) after the introduction of the health programme. Each 1% increase in the proportion of individuals without gallbladder since 10 years was associated with a 0.73% decrease in GBC mortality (95% CI -1.05% to -0.38%), but the negative correlation was limited to women, southern Chile and age over 60. We also found decreasing mortality rates for extrahepatic bile duct, liver, oesophageal and stomach cancer with increasing proportions of individuals without gallbladder. To conclude, 12 years after its inception, the Chilean cholecystectomy programme has markedly and heterogeneously changed cholecystectomy rates. Results based on aggregate data indicate a negative correlation between the proportion of individuals without gallbladder and mortality due to gallbladder and other digestive cancers, which requires validation using individual-level longitudinal data to reduce the potential impact of ecological bias.

Influence of lipid and metabolite profiles of mitochondrial fraction on pH and color stability of longissimus lumborum muscle with different ultimate beef pH.

This study aimed to explore the differences in the lipidome and mitochondrial fraction metabolome of Nellore cattle meat in different ranges of ultimate pH (pHu) normal (≤5.79), intermediate (5.80 to 6.19) and high (≥ 6.20) after 3- and 21-d postmortem. Instrumental color, myoglobin redox state, oxygen consumption, and metmyoglobin-reducing activity were measured during storage. A total of 472 lipids and 22 mitochondrial fraction metabolites were identified. Beef with high pHu showed positive regulation of ceramides involved in apoptosis and negative regulation of lipid classes related to membrane permeability and stability. In addition, lower carnitine content was noted in high-pHu beef than in normal-pHu beef. Acylcarnitines, phosphatidylinositol, and IMP showed upregulation in beef with intermediate pHu, indicating changes mainly related to energy, purine and pyruvate metabolism. Aging time impacted on the lipid content and metabolites involved in different metabolic pathways. These results provided new insights into beef's mitochondrial fraction lipid and metabolic profile with different pHu. In addition, beef with intermediate pHu differs from beef with high pHu due to changes in energy metabolism.

Prevalência de Influenza A, vírus sincicial respiratório eSARS-CoV-2 em pacientes com síndrome respiratóriaaguda grave em Passo Fundo, Rio Grande do Sul / Prevalence of Influenza A, respiratory syncytial virus andSARS-CoV-2 in patients with severe acute respiratorysyndrome in Passo Fundo, Rio Grande do Sul

A síndrome respiratória aguda grave (SRAG) é caracterizada por sintomas de febre alta, tosse e dispneia, e, na maioria dos casos, relacionada a uma quantidade reduzida de agentes infecciosos. O objetivo foi avaliar a prevalência dos vírus respiratórios Influenza A (FluA), vírus sincicial respiratório (RSV) e do novo coronavírus (SARS-CoV-2) em pacientes com internação hospitalar por SRAG. Estudo transversal, com pacientes em internação hospitalar com SRAG entre novembro de 2021 e maio de 2022. Dados sociodemográficos e clínicos e amostras da nasofaringe foram coletados/as, as quais foram submetidas à extração de RNA e testadas quanto à positividade para Influenza A, RSV e SARS-CoV-2 por meio da técnica de PCR em tempo real pelo método SYBR Green. Foram incluídos 42 pacientes, sendo 59,5% do sexo feminino, 57,1% idosos, 54,8% com ensino fundamental. A maior parte dos pacientes reportou hábito tabagista prévio ou atual (54,8%), não etilista (73,8%) e 83,3% deles apresentavam alguma comorbidade, sendo hipertensão arterial sistêmica e diabetes mellitus tipo 2 as mais prevalentes. Um total de 10,5% dos pacientes testou positivo para FluA, nenhuma amostra positiva para RSV e 76,3% positivos para SARS-CoV-2. Na população estudada, SRAG com agravo hospitalar foi observado em maior proporção, em mulheres, idosos e pessoas com comorbidades, embora sem significância estatística, sendo o novo coronavírus o agente etiológico mais relacionado, o que evidencia a patogenicidade desse agente e suas consequências ainda são evidentes após quase 2 anos de período pandêmico.

Severe acute respiratory syndrome (SARS) is characterized by symptoms of high fever, cough and dyspnea, and is in most cases related to a reduced amount of infectious agents. The objective was to assess the prevalence of respiratory viruses Influenza A (FluA), respiratory syncytial virus (RSV) and the new coronavirus (SARS-CoV-2) in patients hospitalized for SARS. Cross-sectional study, with patients hospitalized with SARS between November 2021 and May 2022. Sociodemographic and clinical data and nasopharyngeal samples were collected, which were subjected to RNA extraction and tested for positivity for Influenza A, RSV and SARS-CoV-2 using the real-time PCR technique using the SYBR Green method. 42 patients were included, 59.5% female, 57.1% elderly, 54.8% with primary education. Most patients reported previous or current smoking habits (54.8%), non-drinkers (73.8) and 83.3% of them had some comorbidity, with systemic arterial hypertension and type 2 diabetes mellitus being the most prevalent. A total of 10.5% of patients tested positive for FluA, no samples positive for RSV, and 76.3% positive for SARS-CoV-2. In the studied population, SARS with hospital injury was observed more frequently in women and the elderly, with associated comorbidities, with the new coronavirus being the most related etiological agent, which shows, although not statistically significant, that the pathogenicity of this agent and its consequences are still evident after almost 2 years of period pandemic.

Incidência de obstrução em cateteres periféricos intravenosos em adultos e fatores relacionados / Incidence of obstruction in peripheral intravenous catheters in adults and related factors / Incidencia de obstrucción en catéteres intravenosos periféricos en adultos y factores relacionados

Objetivo: analisar a incidência e os fatores relacionados à obstrução de cateter intravenoso periférico em adultos hospitalizados. Método: uma coorte prospectiva, realizada a partir da observação de 203 cateteres, entre fevereiro de 2019 e maio de 2020, em hospital público e de ensino brasileiro. Foram consideradas variáveis clínicas e do cateter. Os dados foram analisados descritivamente e por estatística inferencial. Resultados: o tempo de permanência variou entre um a 15 dias e a obstrução ocorreu em 7,5% das observações. Houve aumento do risco de obstrução em relação ao sexo (RR=0,49 / p=0,186), à idade (RR=1,20/ p=0,732), aos cateteres de maior calibre (RR=0,53/ p=0,250), à inserção no dorso da mão até antebraço (RR=2,33/ p=0,114) e ao tempo do cateter in situ (RR=033/ p=0,433). Conclusão: O cuidado diário e observação do cateter intravenoso periférico são importantes para minimizar o surgimento de complicações locais e sistêmicas e manter a patência do dispositivo.

Objective: to analyze the incidence and factors related to peripheral intravenous catheter obstruction in hospitalized adults. Method: a prospective cohort, based on the observation of 203 catheters, between February 2019 and May 2020, in a Brazilian public teaching hospital. Clinical and catheter variables were taken into account. The data was analyzed descriptively and using inferential statistics. Results: the length of stay ranged from one to 15 days and obstruction occurred in 7.5% of the observations. There was an increased obstruction risk in relation to gender (RR=0.49 / p=0.186), age (RR=1.20/ p=0.732), larger catheters (RR=0.53/ p=0.250), insertion in the back of the hand up to the forearm (RR=2.33/ p=0.114) and the time length the catheter was in situ (RR=033/ p=0.433). Conclusion: Daily care and observation of the peripheral intravenous catheter is important to minimize the appearance of local and systemic complications and maintain the patency of the device.

Objetivo: analizar la incidencia y los factores relacionados con la obstrucción del catéter intravenoso periférico en adultos hospitalizados. Método: cohorte prospectiva, realizada mediante la observación de 203 catéteres, entre febrero de 2019 y mayo de 2020, en un hospital escuela público brasileño. Se consideraron variables clínicas y del catéter. Los datos se analizaron de forma descriptiva y mediante estadística inferencial. Resultados: el tiempo de permanencia varió entre uno y 15 días y la obstrucción ocurrió en el 7,5% de las observaciones. Hubo mayor riesgo de obstrucción en relación con el sexo (RR=0,49 / p=0,186), la edad (RR=1,20 / p=0,732), los catéteres de mayor calibre (RR=0,53 / p= 0,250), la inserción en el dorso de la mano hasta el antebrazo (RR=2,33/ p=0,114) y el tiempo del catéter in situ (RR=033/ p=0,433). Conclusión: el cuidado diario y la observación del catéter intravenoso periférico son importantes para minimizar la aparición de complicaciones locales y sistémicas y mantener la permeabilidad del dispositivo.

Cognitive and motor improvement by tummy time practice in preemies from low-income settings: a randomized clinical trial.

Introduction: Early intervention and parental education for preemies are limited in some low-income countries. Thus, this study aimed to assess whether daily tummy time (TT) associated with usual care (UC) enhances motor and cognitive development in preemies from low-income countries. The main and secondary aim was to assess prone head elevation (PHE) and motor and cognitive functions, respectively. Methods: Thirty-one preemies with a mean gestational age at birth of 33.3 ± 1.6 weeks and weighing <2,500 g were included and 18 completed all assessments. Parents from the TT group were asked to perform TT with their preemies for at least 20 min during daily activities and playtime. Motor and cognitive functions were assessed by a blinded examiner using the motor and cognitive composite scores of Bayley-III after two and four months of corrected age. PHE was given by the angle from the tragus of the ear to the supporting surface of the infant; measurements were obtained using the Kinovea® software at baseline, after two, three, and four months of corrected age. Results: The Bayley-III motor composite score of the TT group was higher than the UC group after two and four months of corrected age. The PHE angle increased over time but did not differ between groups. Discussion: Nevertheless, TT expanded the perspective of preemies to explore their bodies and environment, favoring the stimuli for motor and cognitive patterns. The loss of participants (31%) was one of the limitations of the study, illustrating the challenge of providing continued early interventions to preemies from low-income countries. In this sense, TT practice is a home intervention that may improve motor and cognitive function of preemies immediately after hospital discharge.

Time series analysis of comprehensive maternal deaths in Brazil during the COVID-19 pandemic.

The effects of the COVID-19 pandemic on comprehensive maternal deaths in Brazil have not been fully explored. Using publicly available data from the Brazilian Mortality Information (SIM) and Information System on Live Births (SINASC) databases, we used two complementary forecasting models to predict estimates of maternal mortality ratios using maternal deaths (MMR) and comprehensive maternal deaths (MMRc) in the years 2020 and 2021 based on data from 2008 to 2019. We calculated national and regional standardized mortality ratio estimates for maternal deaths (SMR) and comprehensive maternal deaths (SMRc) for 2020 and 2021. The observed MMRc in 2021 was more than double the predicted MMRc based on the Holt-Winters and autoregressive integrated moving average models (127.12 versus 60.89 and 59.12 per 100,000 live births, respectively). We found persisting sub-national variation in comprehensive maternal mortality: SMRc ranged from 1.74 (95% confidence interval [CI] 1.64, 1.86) in the Northeast to 2.70 (95% CI 2.45, 2.96) in the South in 2021. The observed national estimates for comprehensive maternal deaths in 2021 were the highest in Brazil in the past three decades. Increased resources for prenatal care, maternal health, and postpartum care may be needed to reverse the national trend in comprehensive maternal deaths.

Oral glucose tolerance test curve shape in Mexican children and adolescents with and without obesity.

OBJECTIVES: Mexican children with obesity are at a higher risk of developing type 2 diabetes mellitus (T2DM). The aim of the study was to compare oral glucose tolerance test (OGTT) characteristics: time of peak glucose, glucose level ≥155 mg/dL at 1 h, presence of metabolic syndrome (MetS), sensitivity, secretion, and oral disposition index (oDI) in children with and without obesity, according to oral glucose tolerance curve shape: monophasic or biphasic. METHODS: Cross-sectional study including 143 children. Groups were divided into (a) obese: biphasic (B-Ob) (n=55) and monophasic (M-Ob) (n=50), (b) without obesity: biphasic (B-NonOb) (n=20) and monophasic (M-NonOb) (n=18). RESULTS: Late glucose peak was more frequent in the M-Ob group (p<0.001). Glucose levels ≥155 mg/dL and MetS were more frequent in the M-Ob group but did not show significance. The groups with obesity (biphasic and monophasic) had higher indices of insulin resistance and insulin secretion compared to the nonobese groups (biphasic and monophasic) (p<0.001). AUC glucose was higher in the M-Ob group (p<0.05), and AUC insulin was higher in the M-NonOb group. oDI (Matsuda) was significantly lower in the M-Ob group compared to the other groups (p<0.001), and oDI-HOMA IR was higher in M-NonOb group (p=0.03). CONCLUSIONS: All OGTT parameters could help to identify Mexican children at increased risk of developing T2DM, not only fasting plasma glucose and 2 h glucose. M-Ob in non-T2DM Mexican children reflects an early defect in glucose metabolism. Higher level of IR indexes in M-NonOb vs. B-NonOb could indicate an increased risk for T2DM of genetic origin.

Linseed oil supplementation alters milk fatty acid profile, mitigates heat stress, and improves summer milk yield in grazing dairy cows.

Dietary supplementation of fat can be an important source of energy to compensate for the reduction in dry matter intake in dairy cows during heat stress periods. Studies have reported that supplementing dairy cow diets with linseed oil (LO) can increase milk yield and enhance the levels of beneficial fatty acids, such as omega-3 fatty acids, in the milk. The objective of this research was to evaluate the effect of LO supplementation on milk fatty acids profile, milk yield and composition, and physiological parameters of grazing cows. The study was conducted in two seasons, one in spring and one in summer. A 2 × 2 Latin square design was used in each experiment. Twelve Holstein and crossbred Holstein x Jersey cows were involved in each season. Cows were divided into two groups: control (TC) with no supplementation and treatment (TL) supplemented with 400 g/day of LO. The results showed that LO supplementation altered the milk fatty acid profile: decreased concentrations of short and medium-chain fatty acids (C10:0 - C17:1) except for C13:0 and increased concentrations of long-chain fatty acids (C18, C18:1 (both trans and cis isomers), C18:2 (specific conjugated linoleic acid - CLA isomers), and C18:3 n3 (omega-3)). Additionally, milk yield increased by 1.5 l per day during summer in LO-supplemented cows, while milk fat, protein, and casein content decreased. Milk stability increased by 2.2% in the LO-supplemented group. LO-supplemented cows reduced internal body temperature and heart frequency in the afternoon and increased daily rumination time by 20 min. In conclusion, LO supplementation can be an effective strategy to improve the nutritional profile of milk by altering fatty acid composition towards potentially healthier fats, mitigate the negative effects of heat stress on grazing cows during summer, as evidenced by reduced body temperature and heart frequency and increase milk yield.

Breast cancer diagnosis and staging in Chile: A non-randomized survey-based study to assess frequency and delays. / Diagnóstico y etapificación de cáncer de mama en Chile: estudio por encuesta no probabilística de III a IV frecuencia y tiempos.

Introduction: Breast cancer progression involves physiological mechanisms such as metastasis. Delays in diagnosis and treatment increase the risk of mortality and are associated with barriers to healthcare access. In Chile, breast cancer is highly prevalent, and early diagnosis has improved, although disparities in the disease evolution persist. This study characterized diagnostic and staging tests, waiting times, and sociodemographic profiles to identify delays and inequities in care. Methods: Survey study. Using a non-probabilistic sample, a questionnaire was applied in an encrypted platform with prior informed consent. The instrument collected data on requested tests, associated times, staging, and sociodemographic characteristics. These variables were analyzed using descriptive statistics, tests of association, confidence intervals, and comparison tests using bootstrapping. Results: A sample of 263 persons was obtained. The most requested tests were biopsy (99.62%) and blood tests (80.23%). The median number of tests requested was six (Q1:4, Q3:8), with a mean of 5.87 (standard deviation: 2.24). No significant differences were observed in the percentage of persons from whom the total number of examinations were requested according to the studied variables. The day-hour-result intervals ranged from 1 to 365 days. The median day-hour-result of the biopsy was 15 days (Q1:10, Q3:30). People between 40 and 49 years old, non-residents of the capital city, belonging to income quintile I, with high school education, from the public health system, with late-stage diagnosis had higher median day-hour-result in biopsy. There was no significant difference in the number of requested tests according to staging (I and II, or III and IV). Conclusions: Biopsy in Chile is the test of choice for diagnostic confirmation in breast cancer. Other tests are requested regardless of the diagnosis stage, contrary to the recommendations of clinical guidelines. Cancer prognosis is crucial, especially in countries with greater inequalities.

Introducción: La progresión del cáncer de mama involucra mecanismos fisiológicos como metástasis. Los retrasos en diagnóstico y tratamiento aumentan el riesgo de mortalidad y se asocian a barreras de acceso a la salud. En Chile, el cáncer de mama es altamente prevalente y su diagnóstico temprano ha mejorado, aunque persisten disparidades en el proceso de enfermedad. Este estudio caracterizó exámenes de diagnóstico y etapificación, tiempos de espera y perfiles sociodemográficos para identificar demoras e inequidades en la atención. Métodos: Estudio de encuesta. Utilizando una muestra no probabilística, se aplicó un cuestionario en plataforma encriptada previo consentimiento informado. En el instrumento se recogieron datos de exámenes solicitados, tiempos asociados, etapificación y características sociodemográficas. Estas variables fueron analizadas utilizando estadística descriptiva, test de asociación, intervalos de confianza y test de comparación utilizando . Resultados: Se logró una muestra de 263 personas. Los exámenes más solicitados fueron biopsia (99,62%) y exámenes de sangre (80,23%). La mediana de exámenes solicitados fue de 6 (Q1:4, Q3:8), con media 5,87 (desviación estándar: 2,24). No se observaron diferencias significativas en el porcentaje de personas a quienes se solicitó la totalidad de exámenes según variables estudiadas. Los intervalos día-hora-resultado oscilaron entre 1 y 365 días. La mediana día-hora-resultado de la biopsia fue de 15 días (Q1:10, Q3:30). Las personas entre 40 y 49 años, no residentes de la capital, pertenecientes al quintil I de ingreso, con educación media, del sistema público de salud, con diagnóstico en etapa tardía presentaron mayores medianas de día-hora-resultado en biopsia. No hubo diferencia significativa en la cantidad de exámenes solicitados según etapificación (I a II y III a IV). Conclusiones: La biopsia en Chile es el examen de elección para la confirmación diagnóstica en cáncer de mama. Otros exámenes son solicitados independientemente de la etapa del diagnóstico, existiendo una discordancia con las recomendaciones de la guía clínica. El pronóstico del cáncer es crucial, especialmente en países con mayores inequidades.

Temporal bisection task as a predictor of cognitive deficits.

Evidence suggests that neuropsychiatric symptoms and deficits in attentional control and executive function can impair time discrimination, demonstrating the intricate link between cognitive processes, subjective well-being, and perception of time. However, the relationship between sleep quality and time discrimination remains elusive. This study aimed to understand differences in the temporal bisection task (TBT) performance. We expected that individuals with impaired cognition, executive function, quality of life, or sleep quality would have reduced time sensitivity. At the same time, those with stress, anxiety, or depression would show a shift in the point of subjective equality. Data were collected from 97 female participants (ranging from 20 to 72 years of age) in more than one moment, resulting in 163 measurements used for the analysis. Participants' neuropsychiatric status was assessed using a battery of tests and scales, including the Mini-Mental State Examination (MMSE), the Frontal Assessment Battery (FAB), the World Health Organization Quality of Life Questionnaire (WHOQOL), the Pittsburgh Sleep Quality Index (PSQI), and the Depression, Anxiety, and Stress Scale-21 items (DASS-21). The results showed that attention and executive control significantly affect time discrimination. In addition, the research indicated that better sleep quality is associated with improved time discrimination sensitivity.

Cytoplasmic strings in human blastocysts: hypotheses of their role and implications for embryo selection.

STUDY QUESTION: What are the implications of the presence cytoplasmic strings (Cyt-S) and their quantity and dynamics for the pre-implantation development of human blastocysts? SUMMARY ANSWER: Cyt-S are common in human embryos and are associated with faster blastocyst development, larger expansion, and better morphological quality. WHAT IS KNOWN ALREADY: Cyt-S are dynamic cellular projections connecting inner cell mass and trophectoderm (TE) cells, that can be observed during blastocyst expansion. Their prevalence in human embryos has been estimated to be between 44% and 93%. Data relevant to their clinical implications and role in development are lacking, limited, or controversial. STUDY DESIGN, SIZE, DURATION: Retrospective study conducted at a single IVF center between May 2013 and November 2014 and involving 124 pre-implantation genetic testing for aneuploidy cycles in a time-lapse incubator with ≥1 blastocyst biopsied and vitrified (N = 370 embryos assessed). These cycles resulted in 87 vitrified-warmed single-euploid blastocyst transfers. PARTICIPANTS/MATERIALS, SETTING, METHODS: ICSI, continuous blastocyst culture (Days 5-7), TE biopsy of fully expanded blastocysts without Day 3 zona pellucida drilling, qPCR to assess uniform full-chromosome aneuploidies, and vitrification were all performed. Only vitrified-warmed euploid single-embryo-transfers were conducted. Blastocyst morphological quality was defined according to Gardner's criteria. The AI-based software CHLOE™ (Fairtility) automatically registered timings from time of starting blastulation (tSB) to biopsy (t-biopsy, i.e. blastocyst full-expansion) as hours-post-insemination (hpi), embryo area (including zona pellucida in µm2), and spontaneous blastocyst collapses. One senior embryologist manually annotated Cyt-S presence, quantity, timings, and type (thick cell-to-cell connections and/or threads). All significant associations were confirmed through regression analyses. All couples', cycles', and embryos' main features were also tested for associations with Cyt-S presence, quantity, and dynamics. MAIN RESULTS AND THE ROLE OF CHANCE: About 94.3% of the patients (N = 117/124) had ≥1 embryo with Cyt-S. Out of a total of 370 blastocysts, 55 degenerated between blastulation and full-expansion (N = 55/370, 14.9%). The degeneration rate among embryos with ≥1 Cyt-S was 10.8% (N = 33/304), significantly lower than that of embryos without Cyt-S (33.3%, N = 22/66, P < 0.01). Of the remaining 315 viable blastocysts analyzed, 86% (N = 271/315; P < 0.01) had ≥1 Cyt-S, on average 3.5 ± 2.1 per embryo ranging 1-13. The first Cyt-S per viable embryo appeared at 115.3 ± 12.5 hpi (85.7-157.7), corresponding to 10.5 ± 5.8 h (0.5-31) after tSB. Overall, we analyzed 937 Cyt-S showing a mean duration of 3.8 ± 2.7 h (0.3-20.9). Cyt-S were mostly threads (N = 508/937, 54.2%) or thick cell-to-cell connections becoming threads (N = 382/937, 40.8%) than thick bridges (N = 47/937, 5.0%). The presence and quantity of Cyt-S were significantly associated with developmentally faster (on average 6-12 h faster) and more expanded (on average 2700 µm2-larger blastocyst's area at t-biopsy) embryos. Also, the presence and duration of Cyt-S were associated with better morphology. Lastly, while euploidy rates were comparable between blastocysts with and without Cyt-S, all euploid blastocysts transferred from the latter group failed to implant (N = 10). LIMITATIONS, REASONS FOR CAUTION: Cyt-S presence and dynamics were assessed manually on seven focal planes from video frames recorded every 15 min. The patients included were mostly of advanced maternal age. Only associations could be reported, but no causations/consequences. Lastly, larger datasets are required to better assess Cyt-S associations with clinical outcomes. WIDER IMPLICATIONS OF THE FINDINGS: Cyt-S are common during human blastocyst expansion, suggesting their physiological implication in this process. Their presence, quantity and dynamics mirror embryo viability, and morphological quality, yet their role is still unknown. Future basic science studies are encouraged to finally describe Cyt-S molecular nature and biophysical properties, and Artificial Intelligence tools should aid these studies by incorporating Cyt-S assessment. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: N/A.

Role of histopathological, serological and molecular findings for the early diagnosis of treatment failure in leprosy.

BACKGROUND: Treatment failure (TF) in leprosy following multidrug therapy (MDT) presents a significant challenge. The current World Health Organization (WHO) fixed-duration MDT regimen, based on lesion count, might not be adequate. Leprosy lacks clear-cut objective cure criteria, and the predictive value of post-MDT histopathological findings remains uncertain. This study aims to identify predictive factors for TF among leprosy patients who have completed the WHO-recommended MDT. METHODS: An analysis was conducted on 80 individuals from a national leprosy reference center, comprising 40 TF cases (with a mean relapse at 13.0 months) and 40 controls (with a mean of 113.1 months without disease signs). Various epidemiological and clinical-laboratory parameters were assessed post-MDT. RESULTS: In skin samples, the presence of foamy granuloma (OR = 7.36; 95%CI2.20-24.60; p = 0.0012) and histological bacillary index (hBI) ≥ 1+ (OR = 1.55; 95%CI1. 22-1.99; p = 0.0004) were significantly associated with TF, with odds ratios of 7.36 and 1.55, respectively. Individuals who experienced TF had a mean hBI of 3.02+ (SD ± 2.02), while the control group exhibited a mean hBI of 1.8+ (SD ± 1.88). An hBI ≥ 3 + showed a sensitivity of 73% and a specificity of 78% for TF detection (AUC: 0.75; p = 0.0001). Other histopathological features like epithelioid granulomas, and skin changes did not show significant associations (p > 0.05). Additionally, higher anti-phenolic glycolipid-I (anti-PGL-I) ELISA index (EI) levels were linked to a 1.4-fold increased likelihood for TF (OR = 1.4; 95%CI1.13-1.74; p = 0.0019). A mean EI of 4.48 (SD ± 2.80) was observed, with an EI ≥ 3.95 showing a sensitivity of 79% and a specificity of 59% for TF detection (AUC: 0.74; p = 0.0001). Moreover, the presence of Mycobacterium leprae (M. leprae) DNA in real-time polymerase chain reaction (qPCR) was associated with a 3.43-fold higher likelihood of TF. Multivariate regression analysis indicated that concurrent presentation of neural/perineural lymphocytic infiltrate, foamy granuloma, hBI ≥ 1+, and EI ≥ 1 markedly increased the likelihood of TF by up to 95.41%. CONCLUSION: Persistence of nerve-selective lymphocytic infiltrate, foamy granulomas, and bacilli in skin biopsies, and elevated EI post-MDT, may serve as predictive factors for identifying individuals at higher probability of TF.

Photobiomodulation Modulates Proliferation and Gene Expression Related to Calcium Signaling in Human Osteoblast Cells.

Introduction: Photobiomodulation with low-level laser treatment can enhance bone formation by stimulating the cell division of osteoblasts and increasing the amount of protein deposition, thus encouraging the formation of new bone. The aim of this study was to evaluate the effects of photobiomodulation with a low-level laser on proliferation and gene expression related to calcium signaling in human osteoblasts. Methods: Osteoblastic cell lines of the hFOB1.19 lineage, human osteoblasts, were grown and assigned into two groups, control (C; n=78 cultured wells) and photobiomodulation (L; n=78 cultured wells) with n=6 per day of the experimental period. Cells were cultured (immature at 34 ºC), and after maturation at 37 ºC, group L cells were exposed to laser irradiation with a low-level laser device (gallium and aluminum arsenide), at a wavelength of 808 nm, a power output of 200 mW, and a power density of 200 mW/cm2. The energy delivered to the cells was 37 J/cm2, with a beam area of 0.02 mm2 and an exposure time of 5 seconds. This treatment was applied daily for a period of 13 days. Following this, the number of cells was counted, and RNA was isolated, measured, and then converted into cDNA for further quantification using a comparative Ct method with real-time polymerase chain reaction. The results were then subjected to statistical analysis through a Mann-Whitney test, with a significance level of P<0.05. Results: The cell count in the L group (37.25x10±4±22.02) was statistically higher compared to the control group (22.75x10±4±7.660) with a P value of 0.0259. The values of 2-ΔΔCt for S100A6, plasma membrane calcium ATPase (PMCA), and calmodulin genes indicated hyper-expression on the thirteenth day, while the osteocalcin gene showed hypo-expression. Conclusion: The study suggests that the photobiomodulation mechanism with a low-level laser may regulate gene expression in human osteoblasts in a dose-dependent and cumulative manner.

Nocturnal activity and forestry-urban dispersal of phlebotomine sand flies (Diptera: Psychodidae) from an ecotourism park in Amazonian Brazil.

Phlebotomine sand flies are insects of notorious importance in public health, mainly due to their involvement in the transmission of Leishmania protozoa. Their flight activity occurs predominantly in the twilight/night period, being stimulated mainly by the need to search for food and reproduction. Despite being naturally wild, some species are able to invade anthropized environments. Present work aimed to assess the nocturnal activity and forestry-urban dispersal of phlebotomine sand flies from an ecotourism park in Belém, Amazonian Brazil. The study area comprised a horizontal transect, extending from a forest park to the neighboring urban environment, in Belém. Sampling was conducted with night-operating light traps. Nocturnal activity was assessed through time-set captures with a collection bottle rotator in the forest environment. Dispersal was assessed through captures carried out along the transect, starting from the forest edge (0 m), extending to the urban environment (50-200 m), phlebotomine sand flies were identified. Abundance, richness, diversity, and sampling sufficiency were estimated. Fourteen species were recorded in the surveyed environments, with Nyssomyia antunesi, Trichophoromyia brachipyga, and Trichophoromyia ubiquitalis being the most abundant. Nocturnal activity behavior of Ny. antunesi occurred between 8 PM and 4 AM while that of Th. brachipyga and Th. ubiquitalis occurred between 2 AM and 6 AM. In the urban environment, the 150 m site presented the highest abundance. Bichromomyia flaviscutellata was sampled in all sites. Gravid females of Pressatia choti and Bi. flaviscutellata were sampled in the urban environment. Putative differential activity between the species herein assessed and their urban dispersal observed are worthy of note, adding data for supporting vector surveillance at a local scale.

Quantifying the Transition from Unconscious to Conscious Detection of Temporal Patterns in Vigilance Tasks: A Unique Adaptation of Mackworth's Clock Test.

This study investigates the cognitive mechanisms underlying vigilance and pattern recognition using a novel adaptation of Mackworth's Clock Test. We aimed to quantify the time it takes for temporal patterns detected unconsciously through implicit learning to surface in the conscious mind within a dynamic vigilance task environment. Forty-eight participants detected random and non-disclosed rhythmic anomalous clock hand movements in this setting. Our results indicate significant variability in detection accuracy, reaction times, and the ability to recognize the hidden pattern among participants. Notably, 23% of all participants and 56% of those who consciously reported the pattern exhibited statistically lower reaction times indicative of knowledge of the pattern 40 s before conscious identification. These findings provide valuable insights into the transition from unconscious to conscious detection, highlighting the complexity of sustained attention and pattern recognition. The study's implications extend to designing training programs and tasks for high-stakes professions requiring prolonged vigilance. Future research should further explore the cognitive and neural correlates of these processes and the impact of task complexity on performance.

Correlation between corrected carotid flow time and left ventricular outflow tract velocity-time integral using a novel technique.

BACKGROUND: Transthoracic echocardiography (TTE) is widely used for assessing patients in the intensive care unit, with cardiac output measurement being crucial for hemodynamic monitoring. This is achieved by measuring the velocity-time integral (VTI) of the left ventricular outflow tract (LVOT), which serves as a surrogate of stroke volume. However, conducting TTE in the critical care setting presents several challenges. Our primary objective was to investigate the relationship between carotid corrected flow time (cCFT) and LVOT VTI. Additionally, we aimed to determine the threshold cCFT value that reliably predicts a normal LVOT VTI. MATERIALS AND METHODS: This proof-of-concept study involves a post-hoc analysis from a diagnostic accuracy investigation conducted in a medical-surgical intensive care unit. We included patients admitted to the ICU from December 2021 to January 2022. We used a phased array transducer to measure the cCFT at the left supraclavicular fossa and the LVOT VTI in an apical 5-chamber view. RESULTS: We included 22 patients. The Spearman coefficient between LVOT VTI and cCFT was 0.82 (p < 0.0001). The area under the ROC curve for cCFT to predict LVOT VTI equal to or greater than 17 cm was 0.871 (95% CI 0.660-0.974). A cCFT exceeding 283 ms predicted LVOT VTI equal to or greater than 17 cm with a sensitivity of 93.3% (95% CI 68.1% to 99.8%) and specificity of 85.7% (95% CI 42.1% to 99.6%). CONCLUSION: The cCFT, measured using a novel technique with a phased array transducer, shows a strong correlation with LVOT VTI. Additionally, cCFT predicts a normal LVOT VTI with good sensitivity and specificity in critically ill patients. Larger studies are warranted to validate these findings.

Murine astrovirus (MuAstV) infection in mouse facilities in Brazil: First South American report.

This study aimed to investigate the presence of murine astrovirus (MuAstV) in Brazil. Fecal samples from mice belonging to four Brazilian animal facilities were collected and tested for MuAstV using real-time polymerase chain reaction. Of the 162 samples tested, 38 (23.5%) were positive for MuAstV, 33 (91.7%) of which came from specific-pathogen free colonies. Although most of the samples were obtained from asymptomatic animals, three mice presented diarrheal symptoms, and MuAstV was the only agent detected by molecular assay. Phylogenetic analysis revealed similarities between the MuAstV strains from this study and prototypes from the USA. MuAstV's high prevalence, environmental stability, genetic diversity and potential for persistent infections must be considered when evaluating health monitoring programs for laboratory rodents.

Spatial and temporal analysis of breast cancer mortality in a state in northeastern Brazil.

Breast cancer (BC) is the most common neoplasm, and its global burden has become one of the most important factors jeopardizing the health of the world population, especially women. The aim of this study was to analyze mortality trends and the spatial distribution of BC in women in the capital and state of Sergipe, aiming to contribute to the implementation and improvement of strategies for the prevention and health promotion of women with BC. Trends were calculated using the Joinpoint Regression Program 5.0.2. Spatial analyses were performed using the empirical Bayesian model, thematic maps were created using QGIS 3.10.7 and Moran's I indices were calculated using TerraView 4.2.2. Between 1996 and 2022, 1384 and 3128 BC deaths were recorded in the capital and state of Sergipe, respectively. The mortality trend increased in the age groups of 45-75+ for the state of Sergipe, while in the capital, we observed stability in all age groups. The highest AAPC was 4.6213, with a 95 % confidence interval (2.16; 7.14). Univariate global Moran's I analysis indicated spatial autocorrelation during the study period. A direct relationship was found between mortality rates and the more economically developed regions.

Impact of the I-DECIDED Tool to Improve Peripheral Intravenous Catheter Care in Paediatrics: Interrupted Time-Series Study.

AIM: To determine whether the I-DECIDED assessment and decision tool enhances peripheral intravenous catheter assessment, care and decision-making in paediatrics. DESIGN: Quasi-experimental, interrupted time-series study. METHODS: An interrupted time-series study was conducted in a paediatric inpatient unit at a public teaching hospital in Brazil. The participants were patients aged less than 15 years old with a peripheral intravenous catheter, and their parents or guardians. Data were collected between January and July 2023, encompassing six time points, three pre-intervention and three post-intervention. Evaluation data were based on the I-DECIDED tool, including idle devices, dressings, complications, patient/family awareness, hand hygiene, disinfection and documentation. RESULTS: We conducted 585 peripheral intravenous catheter observations, with 289 in the pre-intervention phase and 296 in the post-intervention phase, inserted in 65 hospitalised children, 30 in the pre-intervention phase and 35 in the post-intervention phase. After the intervention, reductions were observed in the number of idle catheters, substandard dressings and complications. Patients and family members reported an increase in device assessment, hand hygiene and peripheral intravenous catheter disinfection. Additionally, there was an increase in documentation of decision-making performed by nurses and nursing technicians/assistants. CONCLUSION: Implementation of the I-DECIDED assessment and decision tool in a paediatric unit significantly improved the assessment, care and decision-making regarding peripheral intravenous catheters. IMPLICATIONS FOR THE PROFESSION AND/OR PATIENT CARE: Opportunity to enhance practice standards, elevate the quality of care provided to paediatric patients, contribute to improved patient outcomes, advance evidence-based practice in vascular access management and enhance patient experience through increased involvement in care. IMPACT: To influence clinical practice and healthcare policies aimed at improving peripheral intravenous catheter care and patient safety in paediatric settings. PATIENT OR PUBLIC CONTRIBUTION: No patient or public contribution to the design of this study.