Illumination of understudied ciliary kinases.

Cilia are cellular signaling hubs. Given that human kinases are central regulators of signaling, it is not surprising that kinases are key players in cilia biology. In fact, many kinases modulate ciliogenesis, which is the generation of cilia, and distinct ciliary pathways. Several of these kinases are understudied with few publications dedicated to the interrogation of their function. Recent efforts to develop chemical probes for members of the cyclin-dependent kinase like (CDKL), never in mitosis gene A (NIMA) related kinase (NEK), and tau tubulin kinase (TTBK) families either have delivered or are working toward delivery of high-quality chemical tools to characterize the roles that specific kinases play in ciliary processes. A better understanding of ciliary kinases may shed light on whether modulation of these targets will slow or halt disease onset or progression. For example, both understudied human kinases and some that are more well-studied play important ciliary roles in neurons and have been implicated in neurodevelopmental, neurodegenerative, and other neurological diseases. Similarly, subsets of human ciliary kinases are associated with cancer and oncological pathways. Finally, a group of genetic disorders characterized by defects in cilia called ciliopathies have associated gene mutations that impact kinase activity and function. This review highlights both progress related to the understanding of ciliary kinases as well as in chemical inhibitor development for a subset of these kinases. We emphasize known roles of ciliary kinases in diseases of the brain and malignancies and focus on a subset of poorly characterized kinases that regulate ciliary biology.

Viperin (RSAD2) gene expression in peripheral blood mononuclear cells of pregnant crossbred beef cows is altered by Bos indicus genetics.

The expression of interferon (IFN) stimulated genes (ISGs) in lymphocytes has been used for pregnancy diagnosis in cattle. However, among-cow variability has yielded sub-optimal predictive accuracy. We hypothesized that the expression of ISGs (ISG15, OAS1, RSAD2, CLEC3B, and AKR1B1) in early pregnancy varies according to the proportion of Bos indicus (B. indicus) genetics on females. Multiparous cows were classified in three genetic groups, High Angus (HA; n = 45 [0-33% Brahman influence]), Angus-Brahman (AB; n = 30 [34-67%]), and High Brahman (HB; n = 19 [68-100%]) and submitted to a Select-Synch + CIDR protocol. Cows that displayed estrus (n = 94) were artificially inseminated (Day0; D0). On D19, blood samples were collected to obtain peripheral blood mononuclear cells (PBMC) and measure progesterone (P4) concentrations. On D30, pregnancy diagnosis was performed. The expression of RSAD2 in PBMC of pregnant cows was positively related to the proportion of B. indicus genetics of the groups, but not the expression of ISG15 and OAS1. In pregnant cows, the proportion of B. indicus genetics was negatively associated to circulating levels of P4 concentrations. The P4 concentrations were related positively with RSAD2 expression. ROC curve results determined that for cattle with B. indicus genetics lower than 67%, the CLEC3B and AKR1B1 combination was the most accurate option to predict the outcome of pregnancy. In cows with more than 68% of B. indicus genetics, RSAD2 provided the best accuracy. In conclusion, there is a relationship between the proportion of B. indicus genetics and the ISGs gene expression in PBMC during pregnancy.

Ovarian morphology and follicular dynamics associated with ovarian aging in Bos indicus beef cows.

This study characterizes the ovarian volume, antral follicle count (AFC), and ovarian dynamics of cows at different ages. Nellore cows (Bos taurus indicus) were used in two experiments. In Experiment 1, 57 lactating cows ranging in age from 3 to 23 years had their estrous cycle synchronized based on progesterone/estradiol treatments. The ovaries were scanned by ultrasound to record ovarian volume and AFC prior to follicle aspiration (AFC1). A second AFC (AFC2) was performed 5 days later to count only the growing antral follicles. In Experiment 2, six long-lived (14- to 23-year-old) and three young (4- to 8-year-old) non-lactating cows were submitted to daily ovarian scanning ultrasound during an interovulatory interval. Blood samples were collected during the estrous cycle to assess serum progesterone concentration. Data were analyzed using Student's t-test, ANOVA, and regression analysis tests. In Experiment 1, there were more (P < 0.05) antral follicles in AFC1 (31.4 ± 3.5) than in AFC2 (22.6 ± 2.4). In AFC1, the volume of the right ovary (6.03 ± 0.5 cm3) was greater (P < 0.01) than that of the left one (4.53 ± 0.4 cm3), although the AFC did not differ between the two ovaries (15.3 ± 1.8 and 16.3 ± 1.8, respectively). In both AFC1 and AFC2, there was a decline in the number of antral follicles as the age of the cow increased (P < 0.01). Ovarian volume (average of both ovaries) was related (P < 0.01) with AFC (R² = 0.1499) and cow age (R² = 0.0911). In Experiment 2, young and old cows under the age of 20 had a pattern of follicular growth waves, while cows over 20 years old did not have waves of follicular growth. The progesterone profiles and corpus luteum size during the estrous cycle did not differ between the groups. In conclusion, although cows had increased ovarian volume and decreased follicular population as they age, the follicular growth pattern and corpus luteum functionality appear to be unaffected by age.

Pre-estrus progesterone does not affect post-estrus luminal metabolome in cross-bred beef cows.

In brief: The concentration of progesterone through the estrous cycle modulates uterine function to affect the luminal metabolome. This paper reports that the dynamic changes in the bovine uterine luminal metabolome during diestrus are independent of the concentration of progesterone in the previous cycle. Abstract: In cattle, the concentration of sex steroids modulates uterine function, which is reflected in the composition of the luminal metabolome. Ultimately, the uterine luminal metabolome influences embryonic growth and development. Our objectives were (i) to compare the luminal metabolome 4, 7, and 14 days after estrus of cows that were exposed to greater (HP4; n = 16) vs lower (LP4; n = 24) concentrations of progesterone before displaying estrus and ovulating spontaneously and (ii) to identify changes in the luminal concentration of metabolites across these time points. Luminal epithelial cells and fluid were collected using a cytology brush, and gene expression and metabolite concentrations were assessed by RNAseq and targeted mass spectrometry, respectively. Metabolome profile was similar between treatments within each of days 4, 7, and 14 (false discovery rate (FDR): ≥ 0.1). Concentrations of 53 metabolites changed, independent of treatment, across the diestrus. Metabolites were mostly lipids (40 out 53) and the greatest concentrations were at day 14 (FDR: ≤ 0.1). On day 7, the concentration of putrescine and the gene expression of ODC1, PAOX, SLC3A2, and SAT1 increased (P ≤ 0.05). On day 14, the concentration of 3 ceramides, 4 glucosylceramides, and 12 sphingomyelins and the expression of SGMS2 were increased, in addition to the concentration of choline and 20 phosphatidylcholines. Collectively, the post-estrus concentration of luminal metabolites changed dynamically, independent of the concentration of sex steroids on the previous cycle, and the greatest magnitude changes were on day 14 when lipid metabolism was the most enriched pathway.

Hormonal profile prior to luteolysis modulates the uterine luminal transcriptome in the subsequent cycle in beef cross-bred cows†.

Sex steroid concentrations modulate endometrial function and fertility in cattle. Our objective was to compare the post-estrus luminal transcriptome of cows that were exposed to contrasting concentrations of progesterone (P4) before luteolysis that displayed estrus and ovulated spontaneously. Cross-bred beef cows received either (1) a new CIDR and GnRH (day -9; high progesterone treatment; HP4; n = 16) or (2) a previously used CIDR, PGF2α, and GnRH (low progesterone treatment; LP4; n = 24). All cows received PGF2α at CIDR removal (day -2). Ovarian ultrasonography and blood collections were performed on days -9, -2, -0.5, and 0 (day of observed estrus), and days 4, 7, and 14 for measurement of ovarian structures, P4, and estradiol (E2). Luminal epithelial cells were collected using a cytology brush on days 4, 7, and 14 for RNAseq. On day -2, CL area and concentrations of P4 were greater, while on day -0.5, concentrations of E2 were decreased in HP4. Ovarian structures and hormonal concentrations were similar on days 4, 7, or 14 (P > 0.05). There were enriched pathways in HP4 related to activation and signaling of the innate immune system at day 4, downregulation in the network involved in the extracellular matrix remodeling at day 7, and exacerbated inflammatory response as well as differentiation and activation of macrophages at day 14 (Benjamini-Hochberg P-value ≤ 0.05). In conclusion, manipulation of pre-luteolysis sex steroid concentrations altered the post-estrus luminal transcriptome even though all cows showed estrus and ovulated spontaneously.

Expression profile of key genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin or fetal calf serum.

Cumulus cells from cumulus-oocyte complexes (COC) matured in vitro in serum-free medium show high incidence of apoptosis and DNA double-strand breaks (DSB). This study aimed to characterize the transcript expression profile of selected genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin (BSA) or fetal calf serum (FCS). Briefly, bovine cumulus-oocyte complexes were in vitro matured with either, 0.4% BSA or 10% FCS for 3, 6, 12 or 24 h. The total RNA of cumulus cells was used for real-time PCR analysis. Transcript abundance of XRCC6, XRCC5, DNAPK, GAAD45B, TP53BP1, RAD50, RAD52, ATM and BRCA2 target genes changed as the IVM proceeded (P < 0.05). However, an interaction between protein source (FCS or BSA) and time was not detected (P ≥ 0.05). Cumulus cells from COCs matured with BSA presented higher mRNA expression of two genes compared to FCS group: TP53BP1 at 6 h and BRCA1 at 3, 6, 12 and 24 h (P < 0.05). In summary, our results showed for the first time the expression profile of the key genes involved in DSB repair mechanisms in cumulus cells obtained from bovine COCs matured with FCS or BSA. The higher mRNA expression of BRCA1 and TP53BP1 and lower mRNA expression of TNFAIP6 suggests an increase in apoptosis rate and DNA damage in cumulus cells cultured in BSA-supplemented medium and may explain, at least to some extent, the reduced developmental potential of bovine oocytes matured in serum-free medium.

"A gente é invisível pra sociedade": impactos das condições de trabalho na saúde e qualidade de vida em entregadores de comida na pandemia de covid-19 / "We are invisible to society": impacts of working conditions on food delivery workers' health and quality of life during the COVID-19 pandemic

Resumo A pandemia de covid-19 agravou as situações de renda, fome, desemprego e informalidade provocados pela Reforma Trabalhista de 2017 e pelo desmonte de políticas sociais, levando muitos trabalhadores ao ramo da entrega de comida sem direitos ou garantias de proteção. Este trabalho procurou investigar as condições de trabalho e o impacto na saúde e qualidade de vida de entregadores de comida de Curitiba/PR, de acordo com as categorias de entrega, durante a pandemia de covid-19. Foi realizada pesquisa de campo em 10 points de entrega, seguindo a lógica da saturação de discursos, com o auxílio de instrumento de pesquisa com perguntas semiestruturadas. Paradoxalmente, apesar da menção a vários aspectos negativos das condições de trabalho, a qualidade de vida foi percebida como positiva por grande parte dos entregadores, sendo associada à possiblidade de trabalho e retorno financeiro. Os aspectos negativos das condições de trabalho e seus impactos na saúde e qualidade de vida foram sentidos com mais intensidade por entregadores de aplicativo. Esses achados trazem a necessidade de aprofundamento da discussão das condições de trabalho nos novos arranjos contemporâneos e precarizados e seus impactos na saúde dos trabalhadores, a partir de diferentes contextos e categorias de trabalhadores.

Abstract The COVID-19 pandemic aggravated the scenario of low income, hunger, unemployment, and informality generated by the 2017 Labor Reform and the dismantling of social policies, leading many workers to enter the food delivery business that misses labor rights or protection. Thus, this study aimed at investigating how such working conditions impacted food delivery workers' health and quality of life in Curitiba, Brazil, during the pandemic according to delivery categories. Field research, based on saturation of discourse, was conducted in 10 delivery points using a semi-structured instrument. Despite mentioning several negative aspects regarding working conditions, most delivery workers perceived a positive quality of life, mainly associated with the possibility of work and financial return. App-based delivery workers felt more intensely the negative impacts on health and quality of life. These findings point to the need for further discussions on how these new contemporary and precarious labor arrangements impact occupational health in different contexts and categories.

What a 31-yr multibreed herd taught us about the influence of B. indicus genetics on reproductive performance of cows.

Bos taurus × Bos indicus crosses are widespread in tropical and subtropical regions, nonetheless, quantitative information about the influence of B. indicus genetics on the reproductive performance of beef cattle is lacking. Herein, we determined the association between level of B. indicus genetics and reproduction from a 31-yr dataset comprising sequential breeding seasons of the University of Florida multibreed herd (n = 6,503 Angus × Brahman cows). The proportion of B. indicus genetics in this herd is evenly distributed by each 1/32nd or approximately 3-percentage points. From 1989 to 2020, the estrous cycle of cows was synchronized for artificial insemination (AI) based on detected estrus or timed-AI (TAI) using programs based on gonadotropin-releasing hormone and prostaglandin, and progestin/progesterone. All cows were exposed to natural service after AI and approximately 90-d breeding seasons, considering the day of AI as day 0. The proportion of B. indicus genetics of cows was associated negatively with pregnancy per AI, ranging from 51.6% for cows with 0%-19% of B. indicus genetics to 37.4% for cows with 81%-100% of B. indicus genetics. Similar association was found for estrous response at the end of the synchronization protocol, ranging from 66.3% to 38.4%, respectively. This reduced estrous response helped to explain the pregnancy results, once the pregnancy to AI of cows showing estrus was 2.3-fold greater than for those not showing estrus and submitted to TAI. Despite reduced pregnancy per AI, the increase in the proportion of B. indicus genetics of cows was not associated with a reduction in the proportion of pregnant cows at the end of the breeding season. Nevertheless, the interval from entering the breeding season to pregnancy was lengthened as the proportion of B. indicus genetics of cows increased. The median days to pregnancy was extended by 25 when the proportion of B. indicus genetics surpassed 78% compared with less than 20%. Thus, the increase in the proportion of B. indicus genetics of cows was related to a reduction in pregnancy per AI and lengthening the interval to attain pregnancy during the breeding season, but not with the final proportion of pregnant cows. As a result, reproductive management strategies directed specifically to cows with a greater proportion of B. indicus genetics are needed to improve the rate of pregnancy in beef herds.

Cow­calf operations in the tropics and sub-tropics have benefited from the environmental adaptation provided by Bos indicus genetics. However, reproductive performance has been a cause of concern, although poorly quantified. This study characterized how much the B. indicus genetics in crossbred cows influence herd reproduction. We analyzed data from cows with known proportions of Angus and Brahman genetics, from the same crossbred herd, for 31 sequential breeding seasons. The increase in the proportion of B. indicus genetics reduced estrous response and pregnancy per artificial insemination after estrous synchronization, but not the proportion of pregnant cows at the end of the breeding season. Interval from the beginning of the breeding season to pregnancy was extended by 25 d when the proportion of B. indicus genetics surpassed 78%. In conclusion, reproductive management strategies directed specifically to cows with a greater proportion of B. indicus genetics are needed to improve the rate of pregnancy in beef herds.

Comparing the effect of estradiol benzoate and 17ß-estradiol plus progesterone on follicular turnover and development, and pregnancy outcomes in a timed artificial insemination protocol.

The objective of this study was to compare the efficiency of estradiol benzoate (EB) and 17ß-estradiol (E2) associated with progesterone (P4) in a fixed-time artificial insemination (FTAI) protocol. We hypothesized that E2+P4 induces an earlier emergence of a new follicular wave (NFW), improving pre-ovulatory follicle diameter and pregnancy rates to FTAI (P/FTAI). In Exp.1, on Day 0 (D0), all Bos indicus cows (n = 12/group) received an intravaginal P4 device and a dose of PGF2α analogue. On D0, females were randomly assigned to receive EB or E2+P4. On D8.5, P4 intravaginal devices were removed and a dose of PGF2α and EB were administered in all females followed by fixed-timed AI on D10. Between D0 and D10, the dominant follicular growth was determined by ovary ultrasonography exams. On D8.5 and D10 the percentage of color power-Doppler signals in the dominant follicular wall was evaluated. In Exp. 2, 467 females (2-year-old nulliparous [n = 76], primiparous [n = 92] and pluriparous [n = 299]) were subjected to the similar FTAI and assigned to be treated with EB (n = 243) or E2+P4 (n = 224). Pregnancy diagnosis was performed 30 days after FTAI by ultrasonography. The day to emergence of NFW was similar between treatments (EB: 3.7 ± 0.37 vs. E2+P4: 3.3 ± 0.3, P = 0.76). Females treated with E2+P4 presented greater (P = 0.06) follicular growth between the emergence and D9 (1.18 ± 0.07) than those treated with EB (0.97 ± 0.08). There was also a positive effect (P < 0.05) of E2+P4 on diameter of the dominant follicle on D9 (13.0 ± 0.6 vs. 10.9 ± 0.55) and blood perfusion of the follicle wall on D8.5 (49 vs. 40%). There was a treatment by parity category interaction effect on P/FTAI (P < 0.05). Treatment with E2+P4 was advantageous to P/FTAI of primiparous cows (E2+P4: 58% and EB: 30%). However, for nulliparous and pluriparous cows, P/FTAI was similar between treatments (∼50%). In conclusion, in a E2/P4-based protocol for FTAI, E2+P4 is as efficient as EB in inducing new follicular emergence within a similar day range, but it results similar or greater P/FTAI.

Comparison of estradiol benzoate doses for resynchronization of ovulation at 14 days after timed-AI in suckled beef cows.

We aimed to compare the effects of use of 1 or 2 mg estradiol benzoate (EB) associated with an intravaginal progesterone (P4) device for resynchronization of ovulation 14 days after timed-AI (TAI) in suckled beef cows. Nelore cows were submitted to a TAI (D0) and on D14, received an intravaginal P4 device and were randomly assigned to EB-1 group [n = 516] or EB-2 group [n = 510], which that received 1 or 2 mg EB, respectively. Also, cows had the ovaries scanned by ultrasound to detect an active CL on D14. On D22, devices were removed and structural luteolysis was detected by color-Doppler ultrasonography. In cows which underwent luteolysis, the resynchronization protocol was continued and they were submitted to second TAI on D24. Pregnancy diagnosis was performed 30-35 days after first or second TAI. A subgroup [n = 18-19/group] was submitted to daily ovarian ultrasound scanning from D14 to D22. Proportion of cows with an active CL on D14 did not differ (P > 0.1) between EB-1 and EB-2 groups. The proportion of cows with an active CL on D22 and pregnancy per AI (P/AI) at first TAI were greater (P ≤ 0.05) in EB-1 (55% and 51%) than in EB-2 group (48% and 42%). The P/AI at second TAI did not differ (P > 0.1) between EB-1 (47% [106/227]) and EB-2 group (42% [110/259]). Cumulative pregnancy rate was greater in EB-1 (73% [370/508]) than in EB-2 group (64% [322/502]). No difference (P > 0.1) was observed in the proportion of non-pregnant cows with a synchronized follicular wave emergence between EB-1 and EB-2 groups. In conclusion, treatment with either 1 or 2 mg EB associated with an intravaginal P4 device at D14 after TAI are efficient to synchronize a new follicular wave emergence. The decreased P/AI from first TAI observed in the group of cows receiving 2 mg indicates that this dose is not recommended for use in resynchronization programs initiated 14 days after TAI. The use of 1 mg EB associated with a P4 device provides an elevated cumulative pregnancy rate after two TAIs with an interval of 24 days.

Post-transcriptional and Post-translational Modifications of Primary Cilia: How to Fine Tune Your Neuronal Antenna.

Primary cilia direct cellular signaling events during brain development and neuronal differentiation. The primary cilium is a dynamic organelle formed in a multistep process termed ciliogenesis that is tightly coordinated with the cell cycle. Genetic alterations, such as ciliary gene mutations, and epigenetic alterations, such as post-translational modifications and RNA processing of cilia related factors, give rise to human neuronal disorders and brain tumors such as glioblastoma and medulloblastoma. This review discusses the important role of genetics/epigenetics, as well as RNA processing and post-translational modifications in primary cilia function during brain development and cancer formation. We summarize mouse and human studies of ciliogenesis and primary cilia activity in the brain, and detail how cilia maintain neuronal progenitor populations and coordinate neuronal differentiation during development, as well as how cilia control different signaling pathways such as WNT, Sonic Hedgehog (SHH) and PDGF that are critical for neurogenesis. Moreover, we describe how post-translational modifications alter cilia formation and activity during development and carcinogenesis, and the impact of missplicing of ciliary genes leading to ciliopathies and cell cycle alterations. Finally, cilia genetic and epigenetic studies bring to light cellular and molecular mechanisms that underlie neurodevelopmental disorders and brain tumors.

FOXG1 dose tunes cell proliferation dynamics in human forebrain progenitor cells.

Heterozygous loss-of-function mutations in Forkhead box G1 (FOXG1), a uniquely brain-expressed gene, cause microcephaly, seizures, and severe intellectual disability, whereas increased FOXG1 expression is frequently observed in glioblastoma. To investigate the role of FOXG1 in forebrain cell proliferation, we modeled FOXG1 syndrome using cells from three clinically diagnosed cases with two sex-matched healthy parents and one unrelated sex-matched control. Cells with heterozygous FOXG1 loss showed significant reduction in cell proliferation, increased ratio of cells in G0/G1 stage of the cell cycle, and increased frequency of primary cilia. Engineered loss of FOXG1 recapitulated this effect, while isogenic repair of a patient mutation reverted output markers to wild type. An engineered inducible FOXG1 cell line derived from a FOXG1 syndrome case demonstrated that FOXG1 dose-dependently affects all cell proliferation outputs measured. These findings provide strong support for the critical importance of FOXG1 levels in controlling human brain cell growth in health and disease.

Aumento da Incidência de Recidiva Bioquímica após Prostatectomia Radical em um Centro de Formação em Urologia Oncológica no Brasil: Doenças mais Avançadas estão sendo submetidas à Cirurgia? / Increase of the Incidence of Biochemical Recurrence after Radical Prostatectomy in an Uro-Oncology Training Center in Brazil: Are More Advanced Diseases undergoing Surgery? / Aumento de la Incidencia de Recurrentes Bioquímicos Después de Prostatectomía Radical en un Centro de Capacitación en Urología Oncológica en Brasil: ¿Enfermedades más Avanzadas están siendo sometidas a Cirugía?

Introdução: O câncer de próstata é a neoplasia maligna mais incidente em homens, representando 29% dos diagnósticos da doença no Brasil, segundo o Instituto Nacional de Câncer José Alencar Gomes da Silva (INCA). Esse câncer é suspeito em alterações do toque retal e/ ou do nível sérico do antígeno prostático específico (PSA) total, sendo o diagnóstico definitivo feito por estudo histopatológico. Objetivo: Verificar a associação entre parâmetros clínicos e anatomopatológicos após prostatectomia radical com recidiva bioquímica ao longo do seguimento. Método: Estudo retrospectivo observacional dos parâmetros clínicos (idade, PSA inicial, toque retal, classificação histopatológica da International Society of Urological Pathology (ISUP), escala de D'Amico e estádio clínico) e anatomopatológicos (grau ISUP da peça cirúrgica, margens cirúrgicas, extensão extracapsular tumoral e presença de linfonodos acometidos), de 177 pacientes submetidos à prostatectomia radical em serviço de uro-oncologia de junho/2010-maio/2018. Resultados: A recidiva bioquímica ocorreu em 44,1% dos casos no tempo de seguimento médio de 34,9 meses. A análise univariada demonstrou PSA inicial >9 ng/mL, toque retal alterado, classificação patológica ISUP 4 e 5, risco D'Amico alto e estágio clínico TNM T3 como fatores diretamente associados à recidiva bioquímica. As margens cirúrgicas foram positivas em 46,3%; em 47,7%, identificou-se extensão extraprostática tumoral. Linfonodos positivos em 10,9% e vesículas seminais comprometidas ocorreram em 21,8%. Conclusão: Fatores clínico-patológicos podem ser preditores de recidiva bioquímica. Nesses casos, foi identificado padrão clínico pré-tratamento supostamente mais agressivo em comparação à literatura em geral. Além disso, deve-se considerar a curva de aprendizado dos cirurgiões em formação no serviço, o que pode resultar em maiores taxas de margens cirúrgicas positivas

Introduction: Prostate cancer is the most common cancer in men representing 29% of diagnoses of the disease in Brazil according to the National Cancer Institute José Alencar Gomes da Silva (INCA). If digital rectal examination presents alterations and/or altered serum level of prostate-specific antigen (PSA) total is detected, there is suspicion of prostate cancer, but the definitive diagnosis occurs only with histopathological study. Objective: To correlate clinical and pathological parameters after radical prostatectomy with biochemical recurrence during follow-up. Method: Retrospective observational study of clinical parameters (age, initial PSA, digital rectal examination, histopathological classification of the International Society of Urological Pathology (ISUP), D'Amico scale and clinical stage) and pathological (ISUP degree of the surgical specimen, surgical margins, extracapsular tumor extension and presence of positive lymph nodes) of 177 patients who underwent radical prostatectomy in an uro-oncology service from June 2010 to May 2018. Results: Biochemical recurrence occurred in 44.1% of the cases within a mean follow-up time of 34.9 months. Univariate analysis showed that baseline PSA>9 ng/mL, altered rectal examination, pathological ISUP classification 4 and 5, high D'Amico risk, and clinical TNM stage T3 are risk factors for biochemical recurrence. Surgical margins were positive in 46.3%, and in 47.7% extracapsular extension was identified. Positive lymph nodes were detected in 10.9% and positive seminal vesicles occurred in 21.8%. Conclusion: Clinical and pathological factors can be predictors of biochemical recurrence. In these cases, it was identified a more aggressive clinical pattern than the literature in general. In addition, it should be considered the learning curve of surgeons in training at the service, which can result in higher rates of positive surgical margins

Introducción: El cáncer de próstata es lo más incidente en hombres representando 29% de los diagnósticos de enfermedades en Brasil según Instituto Nacional del Cáncer José Alencar Gomes da Silva (INCA), se sospecha en tacto rectal y/o en el nivel de análisis del antígeno prostático específico (PSA) total alterado, y el diagnóstico definitivo se realiza mediante el estudio histopatológico. Objetivo: Correlacionar los parámetros clínicos y patológicos después de la prostatectomía radical con la recurrencia bioquímica a lo largo del seguimiento. Método: Estudio observacional retrospectivo de parámetros clínicos (edad, PSA inicial, tacto rectal, clasificación histopatológica de la International Society of Urological Pathology (ISUP), escala D'Amico y estadio clínico) y patológicos (grado ISUP de la muestra quirúrgica, márgenes quirúrgicos, extensión capsular tumoral extra y ganglios linfáticos positivos) de 177 pacientes sometidos a prostatectomía radical en servicio de uro-oncología de junio/2010-mayo/2018. Resultados: La recurrencia bioquímica ocurrió en el 44,1% de los casos en un tiempo de seguimiento promedio de 34,9 meses. El análisis univariado demostró que el PSA inicial >9 ng/mL, alteración del tacto rectal, clasificación patológica ISUP 4 y 5, alto riesgo de D'Amico y estadificación TNM clínico T3 como factores de riesgo para recurrencia bioquímica. Los márgenes quirúrgicos fueron positivos en el 46,3%, y en el 47,7% se identificó una extensión extra capsular adicional. Los ganglios linfáticos positivos fueron detectados en 10,9% y las vesículas seminales positivas ocurrieron en el 21,8%. Conclusión: Los factores clínicos y patológicos pueden ser predictores de recurrencia bioquímica. En estos casos, fue identificado un patrón más agresivo que la literatura en general. Además, se debe considerar la curva de aprendizaje de los cirujanos en formación en el servicio, lo que puede resultar en mayores tasas de márgenes quirúrgicos positivos

Peri-estrus ovarian, uterine, and hormonal variables determine the uterine luminal fluid metabolome in beef heifers.

In cattle, uterine luminal fluid (ULF) is the main source of molecules that support embryo development and survival during the peri-implantation period. Our overarching hypothesis is that peri-estrus changes in uterine function, including ULF accumulation and absorption, are uneven among individuals, and affect ULF composition and fertility. Our objectives were (1) to characterize temporal and spatial changes in ULF volume, endometrial and luteal blood perfusion, endometrial and luteal size, and circulating progesterone concentrations during the peri-estrus period in beef heifers and (2) to associate such changes with the metabolite composition in the ULF, 4 days after estrus (d 0). Fourteen Bos indicus heifer that presented a PGF2α responsive CL received 500 µg PGF2α analog i.m. and were examined daily by rectal B-mode and pulse-wave color-Doppler ultrasonography until the fifth day after estrus (d 5). The composition of the ULF was analyzed by targeted mass spectrometry on d 4. Multivariate analyses clustered heifers according to ovarian, uterine, and hormonal variables in clusters A (n = 5) and B (n = 8 heifers). Concentrations of Pro, Ala, Leu, Gly, Val, Lys, Ile, Phe, Asp, Orn, Tyr, Arg, Trp, Suc, Cit, ADMA, the sum of essential Amino Acids (AA), sum of nonessential AA, sum of aromatic AA, and total AA were greater in cluster A (FDR ≤ 0.05). ULF volume dynamics and uterine, ovarian, and hormonal variables during the peri-estrus period presented a concerted variation among heifers within clusters, which was associated with the ULF composition 4 days after estrus.

Applied use of interferon-tau stimulated genes expression in polymorphonuclear cells to detect pregnancy compared to other early predictors in beef cattle.

We aimed to evaluate the accuracy of Interferon-tau stimulated genes (ISG) abundance in peripheral blood polymorphonuclear cells (PMNs) on D20 after fixed-time artificial insemination (FTAI; D0) as a pregnancy diagnosis method against CL evaluation by Doppler ultrasonography and progesterone (P4) concentrations on D20, as well as Pregnancy Associated Glycoproteins (PAG) concentrations on D25. Additionally, we evaluated the potential of ISG abundance in PMNs as pregnancy loss predictors. Nelore heifers (n = 103) and cows (n = 144) underwent estrous synchronization and were artificially inseminated on D0. Pregnancy was diagnosed by B-mode ultrasonography on D30 and D70, and after the final diagnosis, females were classified in four groups: Pregnant; Non-pregnant; Functional CL on D20 but non-pregnant (CL-NP) and Pregnancy loss between D30 and D70 (PL). After determining cutoff values, the Sensitivity (SE), Specificity (SP), Positive Predictive Value (PPV), Negative Predictive Value (NPV) and Accuracy (ACC) were determined for each method. All methods were classified as significant (P < 0.05) predictors of pregnancy. Both ISG expression and PAG concentrations were greater (P < 0.05) in pregnant females than in non-pregnant and CL-NP females but did not differ (P > 0.05) from the PL group. ISG15 expression was greater (P < 0.05) in heifers than in cows, but this difference was not found in OAS1 expression and PAG concentrations. All the methods evaluated were proven to be adequate predictors of pregnancy, but greater accuracies were obtained through PAG concentrations and Doppler-US, due to the decreased number of false positive and false negative results.

The pre-hatching bovine embryo transforms the uterine luminal metabolite composition in vivo.

In cattle, conceptus development after elongation relies on well-characterized, paracrine interactions with the hosting maternal reproductive tract. However, it was unrecognized previously that the pre-hatching, pre-implantation bovine embryo also engages in biochemical signalling with the maternal uterus. Our recent work showed that the embryo modified the endometrial transcriptome in vivo. Here, we hypothesized that the embryo modulates the biochemical composition of the uterine luminal fluid (ULF) in the most cranial portion of the uterine horn ipsilateral to the corpus luteum. Endometrial samples and ULF were collected post-mortem from sham-inseminated cows and from cows inseminated and detected pregnant 7 days after oestrus. We used quantitative mass spectrometry to demonstrate that the pre-hatching embryo changes ULF composition in vivo. Embryo-induced modulation included an increase in concentrations of lipoxygenase-derived metabolites [12(S)-HETE, 15(S)-HETE] and a decrease in the concentrations of amino acids (glycine), biogenic amines (sarcosine), acylcarnitines and phospholipids. The changed composition of the ULF could be due to secretion or depletion of specific molecules, executed by either the embryo or the endometrium, but initiated by signals coming from the embryo. This study provides the basis for further understanding embryo-initiated modulation of the uterine milieu. Early embryonic signalling may be necessary to guarantee optimal development and successful establishment of pregnancy in cattle.

Ultrasonography-accessed luteal size endpoint that most closely associates with circulating progesterone during the estrous cycle and early pregnancy in beef cows.

The aim was to evaluate the associations between circulating P4 concentrations, corpus luteum (CL) size (diameter, area or volume) and blood perfusion (BP) in cows. In Experiment 1, Pearson's correlations (P < 0.05) with P4 concentrations were observed during CL development (D8) for total area (TA; r = 0.76), luteal area (ACL; r = 0.72), total and luteal diameter (TD and DCL respectively; r = 0.46). During mid-late diestrus, there was a positive correlation (P < 0.05) only at D15 with TA and ACL (r > 0.60), TD, total volume (TV) and luteal volume (VCL; r > 0.434). During luteal regression, the correlation was only observed at D18 for ACL (r = 0.478) and D20 with several variables. In Experiment 2, CL weight and ACL had the greatest correlation with P4 (r > 0.6). In Experiment 3, TA and ACL were the variables that were most closely correlated with serum P4 concentrations at D7 in recipient cows. Correlation coefficients were greater for luteal measurements when there were compact compared with cavitary CLs. In Experiment 4, there was no correlation (P > 0.05) between P4 and any of the variables measured on D4 and D7 in recipient cows detected in estrus. On D18 to D20, all CL characteristics were correlated (P < 0.05) with plasma P4, and luteal BP and BP area were more closely (P < 0.05) correlated than ACL. In conclusion, CL perimeter area measurements had the greatest association with luteal function during CL development; whereas for BP there was a greater correlation with P4 than luteal size during luteolysis.

Reproductive performance of Bos indicus beef cows treated with different doses of equine chorionic gonadotropin at the end of a progesterone-estrogen based protocol for fixed-time artificial insemination.

Two experiments were performed to evaluate the reproductive performance of zebu beef cows treated with different doses of eCG at the end of a progesterone (P4)/estrogen based protocol for timed artificial insemination (TAI). In Experiment 1, suckling Bos indicus Nelore cows (n = 261) received, on day 0, a progesterone (P4) intravaginal device (PD) and an injection of 1 mg estradiol benzoate (EB). On day 8, the PD was removed, 500 µg of cloprostenol was injected, and cows were assigned to one of the following groups: Control (no treatment), 300 (300 IU of eCG), 600 (600 IU of eCG), and 900 (900 IU of eCG). On day 9, all cows received 1 mg EB and TAI performed 54-56 h after cloprostenol injection. A pregnancy diagnosis was done by ultrasound scanning 40 days after TAI, and the number of fetuses and calves was recorded at pregnancy diagnosis and at birth. More cows treated with eCG displayed estrus within 48 h after removal of the PD (42.3% vs. 11.6%, P < 0.01), and ovulated more than one follicle (42%, 58/138 vs. 1.8%, 1/54; P < 0.01). This effect on ovulation rate was dose dependent (P < 0.05). The pregnancy rate was affected only by cow parity (primiparous, 25.3% vs. multiparous, 48.9%; P < 0.01). Twin pregnancy was higher (P < 0.01) in cows treated with eCG (42%, 58/138) than controls (0%, 0/54). However, few cows (33.3%) were able to keep both fetuses intact until birth. For evaluation of ovarian characteristics by B-mode and Doppler ultrasonography, 43 Nelore cows were submitted In Experiment 2 to the same four groups described in Experiment 1. Although no difference (P > 0.1) was observed for size and blood perfusion in the pre-ovulatory follicles, corpus luteum was larger and with greater blood perfusion (P < 0.05) in eCG-treated cows. In conclusion, eCG increased the number of double/multiple ovulations in a dose-dependent manner, induced larger and more vascularized corpora lutea, but did not affect the fertility of cyclic or anestrous cows. Although eCG results in twin pregnancies, most of cows underwet embryo/fetus loss and birth a single calf.

Education, practical training and professional development for public health practitioners: a scoping review of the literature and insights for sustainable food system capacity-building.

OBJECTIVE: Noting the upstream positioning of sustainable food systems (SFS) to multiple global crises, the present review described examples of emerging and promising practices to support SFS-oriented education, practical training (PT) and continuing professional development (CPD) among trainees and public health practitioners (PHP). A secondary objective was to compile the evidence into practical considerations for educators, supervising practitioners and professional associations. DESIGN: A scoping review of the literature published between 2007 and 2017 was conducted in May 2017 using four databases: CINAHL, MEDLINE, Scopus and HSSA, along with bibliography hand-searching and expert consultation. Articles were screened for relevance and specificity by independent raters. RESULTS: Nineteen articles were included for analysis. Two-thirds of the articles related to dietitians and public health nutritionists. Emerging practices included curriculum-based considerations, incorporation of 'sustainability' within professional competencies and self-reflection related to SFS. Descriptions of SFS-related education, PT and CPD practices appeared largely in the literature from developed countries. Articles converged on the need for ecosystems, food systems and sustainability considerations within and across practice to support current and future practitioners. CONCLUSIONS: There is growing interest in SFS but guidance to support educators and preceptors is lacking. Updates to dietary guidelines to reflect issues of sustainability are a timely prompt to examine the education, training and development needs of trainees and PHP. Practical examples of emerging practices can empower PHP to promote SFS in all areas of practice. More research is needed to address identified gaps in the literature and to improve SFS-specific education, PT and CPD.

International cooperation for food and nutrition security: Systematization of the participatory, contextualized, and intersectoral educational practices / Cooperação internacional em segurança alimentar e nutricional: sistematização de práticas educativas participativas, contextualizadas e intersetoriais

ABSTRACT The present study systematized the experience gained with the project Construindo capacidades em segurança alimentar e nutricional no Brasil, Canadá e Angola (2004-2010, Building food and nutrition security skills in Brazil, Canada, and Angola), whose objective was to qualify actions that promote food and nutrition security in the three countries using different educational practices. The activities were organized in the following subprojects: (a) online distance learning courses; (b) workshops to train managers, government technicians, representatives of civil society organizations, and social subjects who offered to act as a link between communities; and (c) local pilot projects. The present study reports this experience. The educational practices implemented in the municipalities of Araçuaí (MG), Juazeiro (BA), and Fortaleza (CE) were analyzed based on systematized information in the project reports and activity records (texts and photographs). The analytical reference was based on the concept of food and nutrition education, guided by the fundamentals of Popular Education and Paulo Freire; on the concept of food and nutrition security; and on the following analytical dimensions: participation, contextualization of educational practices, and intersectoriality. The results evidenced how educational practices contributed to the construction of shared concepts of food and nutrition security from an intersectoral and participatory perspective that values the peculiarities of diet in different socioeconomic and cultural contexts, and highlights daily situations and local traditions. They also expose the limits and potentialities of an experience of this magnitude, conducted from an interdisciplinarity perspective and using participatory methods.

RESUMO O presente estudo consiste em uma sistematização da experiência do projeto Construindo capacidades em segurança alimentar e nutricional no Brasil, Canadá e Angola (2004-2010), que teve como objetivo qualificar as ações promotoras de segurança alimentar e nutricional nos três países por meio de diferentes práticas educativas. As atividades foram organizadas nos seguintes subprojetos: (a) cursos de formação à distância (online); (b) oficinas de qualificação de gestores, técnicos de governos, representantes de organizações da sociedade civil e sujeitos sociais que se dispuseram a atuar como elos entre as comunidades; (c) projetos piloto no nível local. Dessa maneira, o estudo consiste em um relato dessa experiência. Foram analisadas as práticas educativas implementadas nos municípios de Araçuaí (MG), Juazeiro (BA) e Fortaleza (CE) a partir das informações sistematizadas nos relatórios do projeto e registros de atividades (escritos e fotográficos). O referencial analítico baseou-se no conceito de educação alimentar e nutricional, pautado nos fundamentos da Educação Popular e de Paulo Freire, no conceito de segurança alimentar e nutricional e nas seguintes dimensões de análise: participação, contextualização das práticas educativas e intersetorialidade. Os resultados evidenciam como as práticas educativas contribuíram para uma construção compartilhada de concepções sobre Segurança Alimentar e Nutricional, numa perspectiva intersetorial e participativa, ao valorizarem as peculiaridades da alimentação nos diferentes contextos socioeconômicos e culturais e evidenciarem situações cotidianas e tradições locais. Apontam, também, os limites e potencialidades de uma experiência de tal amplitude e diversidade, conduzida sob a ótica da interdisciplinaridade e do trabalho participativo.