Characterization of expression patterns and dynamic relocation of Notch proteins during acrosome reaction of bull spermatozoa.

Notch is a conserved cell-signaling pathway involved in spermatogenesis regulation. This study firstly evaluated the presence, localization patterns, acquisition origin and relation to acrosome reaction of Notch proteins in bull sperm. Western Blot analysis detected all Notch proteins in ejaculated bull sperm, and immunostaining described their specific sperm localization. Recovery of sperm from different segments showed that Notch proteins have testicular origin (NOTCH1, NOTCH2, DLL4), are sequentially acquired during sperm maturation along epididymal transit (NOTCH3, DLL3, JAGGED1-2), or post-ejaculation (DLL1, NOTCH4). Testis NOTCH2 is ubiquitously expressed in all germ-cell lines, whereas DLL4 is expressed in round and elongated spermatids during the Golgi, Cap, Acrosome and Maturation phases. In vitro spontaneous and induced sperm acrosome reaction induce consistent sperm regional relocation of NOTCH2, DLL4 and JAGGED1, and these relocation patterns are significantly associated to sperm acrosome status. NOTCH2 and JAGGED1 are relocated from the head apical to the post-equatorial regions, whereas DLL4 is lost along with the acrosome, evidencing that sperm spatial redistribution of NOTCH2 and JAGGED1 is linked to acrosome reaction onset, whereas DLL4 loss is linked to AR completion. Overall, results prompt for a relevant Notch role in bull sperm acrosome testicular development, epididymal maturation and acrosome reaction.

Chemical reversion of age-related oocyte dysfunction fails to enhance embryo development in a bovine model of postovulatory aging.

PURPOSE: There are no clinical treatments to prevent/revert age-related alterations associated with oocyte competence decline in the context of advanced maternal age. Those alterations have been attributed to oxidative stress and mitochondrial dysfunction. Our study aimed to test the hypothesis that in vitro maturation (IVM) medium supplementation with antioxidants (resveratrol or phloretin) may revert age-related oocyte competence decline. METHODS: Bovine immature oocytes were matured in vitro for 23 h (young) and 30 h (aged). Postovulatory aged oocytes (control group) and embryos obtained after fertilization were examined and compared with oocytes supplemented with either 2 µM of resveratrol or 6 µM phloretin (treatment groups) during IVM. RESULTS: Aged oocytes had a significantly lower mitochondrial mass and proportion of mitochondrial clustered pattern, lower ooplasmic volume, higher ROS, lower sirtuin-1 protein level, and a lower blastocyst rate in comparison to young oocytes, indicating that postovulatory oocytes have a lower quality and developmental competence, thus validating our experimental model. Supplementation of IVM medium with antioxidants prevented the generation of ROS and restored the active mitochondrial mass and pattern characteristic of younger oocytes. Moreover, sirtuin-1 protein levels were also restored but only following incubation with resveratrol. Despite these findings, the blastocyst rate of treatment groups was not significantly different from the control group, indicating that resveratrol and phloretin could not restore the oocyte competence of postovulatory aged oocytes. CONCLUSION: Resveratrol and phloretin can both revert the age-related oxidative stress and mitochondrial dysfunction during postovulatory aging but were insufficient to enhance embryo developmental rates under our experimental conditions.

Immunolocalization of Notch Signaling in Mouse Preimplantation Embryos.

The Notch signaling pathway is an important determinant of cell diversity and identity in most developing embryonic tissues. The pathway components are expressed dynamically, and their function is critical for embryonic survival.This protocol addresses the immunolocalization of Notch pathway components by confocal microscopy.

CROSS-CULTURAL ADAPTATION AND VALIDATION OF THE MONTREAL CHILDREN'S HOSPITAL FEEDING SCALE INTO BRAZILIAN PORTUGUESE.

OBJECTIVE: To cross-culturally adapt and validate the Montreal Children's Hospital Feeding Scale (MCH-FS) into Brazilian Portuguese. METHODS: The MCH-FS, originally validated in Canada, was validated in Brazil as Escala Brasileira de Alimentação Infantil (EBAI) and developed according to the following steps: translation, production of the Brazilian Portuguese version, testing of the original and the Brazilian Portuguese versions, back-translation, analysis by experts and by the developer of the original questionnaire, and application of the final version. The EBAI was applied to 242 parents/caregivers responsible for feeding children from 6 months to 6 years and 11 months of age between February and May 2018, with 174 subjects in the control group and 68 ones in the case group. The psychometric properties evaluated were validity and reliability. RESULTS: In the case group, 79% of children were reported to have feeding difficulties, against 13% in the control group. The EBAI had good internal consistency (Cronbach's alpha=0.79). Using the suggested cutoff point of 45, the raw score discriminated between cases and controls with a sensitivity of 79.4% and specificity of 86.8% (area under the ROC curve=0.87). CONCLUSIONS: The results obtained in the validation process of the EBAI demonstrate that the questionnaire has adequate psychometric properties and, thus, can be used to identify feeding difficulties in Brazilian children from 6 months to 6 years and 11 months of age.

Balanced Notch-Wnt signaling interplay is required for mouse embryo and fetal development.

This study investigated the role of Notch and Wnt cell signaling interplay in the mouse early embryo, and its effects on fetal development. Developmental kinetics was evaluated in embryos in vitro cultured from the 8-16-cell to the hatched blastocyst stage in the presence of signaling inhibitors of Notch (DAPT) and/or Wnt (DKK1). An embryo subset was evaluated for differential cell count and gene transcription of Notch (receptors Notch1-4, ligands Dll1, Dll4, Jagged1-2, effectors Hes1-2) and Wnt (Wnt3a, Lrp6, Gsk3ß, C-myc, Tcf4, ß-catenin) components, E-cadherin and pluripotency and differentiation markers (Sox2, Oct4, Klf4, Cdx2), whereas a second subset was evaluated for implantation ability and development to term following transfer into recipients. Notch and Wnt blockades had significant opposing effects on developmental kinetics - Notch blockade retarded while Wnt blockade fastened development. This evidences that Notch and Wnt regulate the pace of embryo kinetics by respectively speeding and braking development. Blockades significantly changed the transcription profile of Sox2, Oct4, Klf4 and Cdx2, and Notch and double blockades significantly changed embryonic cell numbers and cell ratio. The double blockade induced more severe phenotypes than those expected from the cumulative effects of single blockades. Implantation ability was unaffected, but Notch and double blockades significantly decreased fetal development to term. Compared to control embryos, Notch blockade and Wnt blockade embryos originated, respectively, significantly lighter and heavier fetuses. In conclusion, Notch and Wnt signaling interplay in the regulation of the pace of early embryo kinetics, and their actions at this stage have significant carry-over effects on later fetal development to term.

Cross-cultural adaptation and validation of the montreal children's hospital feeding scale into brazilian portuguese / Adaptação transcultural e validação da montreal children's hospital feeding scale para o português falado no brasil

ABSTRACT Objective: To cross-culturally adapt and validate the Montreal Children's Hospital Feeding Scale (MCH-FS) into Brazilian Portuguese. Methods: The MCH-FS, originally validated in Canada, was validated in Brazil as Escala Brasileira de Alimentação Infantil (EBAI) and developed according to the following steps: translation, production of the Brazilian Portuguese version, testing of the original and the Brazilian Portuguese versions, back-translation, analysis by experts and by the developer of the original questionnaire, and application of the final version. The EBAI was applied to 242 parents/caregivers responsible for feeding children from 6 months to 6 years and 11 months of age between February and May 2018, with 174 subjects in the control group and 68 ones in the case group. The psychometric properties evaluated were validity and reliability. Results: In the case group, 79% of children were reported to have feeding difficulties, against 13% in the control group. The EBAI had good internal consistency (Cronbach's alpha=0.79). Using the suggested cutoff point of 45, the raw score discriminated between cases and controls with a sensitivity of 79.4% and specificity of 86.8% (area under the ROC curve=0.87). Conclusions: The results obtained in the validation process of the EBAI demonstrate that the questionnaire has adequate psychometric properties and, thus, can be used to identify feeding difficulties in Brazilian children from 6 months to 6 years and 11 months of age.

RESUMO Objetivo: Realizar a adaptação transcultural e a validação da escala Montreal Children's Hospital Feeding Scale (MCH-FS) para a língua portuguesa falada no Brasil. Métodos: A MCH-FS, originalmente validada no Canadá, foi validada no Brasil como Escala Brasileira de Alimentação Infantil (EBAI) e desenvolvida a partir das seguintes etapas: tradução, montagem da versão em português brasileiro, teste da versão em inglês e da versão em português brasileiro, retrotradução, análise por experts e autora do questionário original e aplicação da versão final em estudo. A EBAI foi aplicada em 242 pais/cuidadores responsáveis pela alimentação de crianças de seis meses a seis anos e 11 meses de idade no período de fevereiro a maio de 2018, sendo 174 no grupo controle e 68 no grupo dos casos. As propriedades psicométricas avaliadas foram validade e confiabilidade. Resultados: No grupo dos casos, 79% dos pais/cuidadores relataram dificuldades alimentares, e no grupo controle, 13%. A EBAI apresentou boa consistência interna (alfa de Cronbach=0,79). Utilizando-se o ponto de corte sugerido de 45, o escore bruto (raw score) diferenciou casos de controles com sensibilidade de 79,4% e especificidade de 86,8% (área sob a curva ROC=0,87). Conclusões: Os resultados obtidos na validação da EBAI evidenciaram medidas psicométricas adequadas. Portanto, a escala pode ser utilizada na identificação de dificuldades alimentares em crianças brasileiras de seis meses a seis anos e 11 meses idade.

Notch signaling in mouse blastocyst development and hatching.

BACKGROUND: Mammalian early embryo development requires a well-orchestrated interplay of cell signaling pathways. Notch is a major regulatory pathway involved in cell-fate determination in embryonic and adult scenarios. However, the role of Notch in embryonic pre-implantation development is controversial. In particular, Notch role on blastocyst development and hatching remains elusive, and a complete picture of the transcription and expression patterns of Notch components during this time-period is not available. RESULTS: This study provided a comprehensive view on the dynamics of individual embryo gene transcription and protein expression patterns of Notch components (receptors Notch1-4; ligands Dll1 and Dll4, Jagged1-2; and effectors Hes1-2), and their relationship with transcription of gene markers of pluripotency and differentiation (Sox2, Oct4, Klf4, Cdx2) during mouse blastocyst development and hatching. Transcription of Notch1-2, Jagged1-2 and Hes1 was highly prevalent and dynamic along stages of development, whereas transcription of Notch3-4, Dll4 and Hes2 had a low prevalence among embryos. Transcription levels of Notch1, Notch2, Jagged2 and Hes1 correlated with each other and with those of pluripotency and differentiation genes. Gene transcription was associated to protein expression, except for Jagged2, where high transcription levels in all embryos were not translated into protein. Presence of Notch signaling activity was confirmed through nuclear NICD and Hes1 detection, and downregulation of Hes1 transcription following canonical signaling blockade with DAPT. In vitro embryo culture supplementation with Jagged1 had no effect on embryo developmental kinetics. In contrast, supplementation with Jagged2 abolished Jagged1 transcription, downregulated Cdx2 transcription and inhibited blastocyst hatching. Notch signaling blockade by DAPT downregulated transcription of Sox2, and retarded embryo hatching. CONCLUSION: Transcription of Notch genes showed a dynamic pattern along blastocyst development and hatching. Data confirmed Notch signaling activity, and lead to the suggestion that Notch canonical signaling may be operating through Notch1, Notch3, Jagged1 and Hes1. Embryo culture supplementation with Jagged1 and Jagged2 unveiled a possible regulatory effect between Jagged1, Cdx2 and blastocyst hatching. Overall, results indicate that a deregulation in Notch signaling, either by its over or under-activation, affects blastocyst development and hatching.

Spontaneous outer retinal layer recovery in a case of hypertensive choroidopathy secondary to pre-eclampsia: a multimodal evaluation / Recuperação espontânea das camadas externas da retina em um caso de coroidopatia hipertensiva secundária a pré-eclâmpsia: avaliação multimodal

ABSTRACT A 26-year-old woman presented at 28 weeks gestation with hypertensive choroidopathy associated with pre-eclampsia. Fundus photography, fundus autofluorescence, spectral-domain optical coherence tomography (SD-OCT), fluorescein angiography, and indocyanine green angiography were performed in both eyes in the immediate postoperative period. SD-OCT images were obtained before delivery and during a 3-month follow-up. Fundus autofluorescence exhibited patchy hyper- and hypoautofluorescent lesions; fluorescein and indocyanine green angiography revealed areas of choroidal ischemia; and SD-OCT showed disorganization of the outer retinal layers and disruption of the ellipsoid zone. After her blood pressure was stabilized, progressive recovery of the outer retinal layer was monitored on SD-OCT.

RESUMO Uma mulher de 26 anos de idade, com 28 semanas de gestação apresentando coroidopatia hipertensiva associada à pré-eclâmpsia. Retinografia, autofluorescência, tomografia de coerência óptica de domínio espectral, angiofluoresceínografia e angiografia com indocianina verde foram realizadas em ambos os olhos no período pós-operatório imediato do parto. Imagens da tomografia de coerência óptica de domínio espectral foram obtidas antes do parto e durante o seguimento de 3 meses. A autofluorescência apresentou lesões heterogêneas hiper e hipoautofluorescentes, a angiofluoresceínografia e angiografia com indocianina verde revelaram áreas de isquemia de coroide, e a tomografia de coerência óptica de domínio espectral apresentou desorganização das camadas externas da retina e interrupção da zona elipsóide. Após a estabilização da pressão sanguínea, a recuperação progressiva da camada externa da retina foi monitorada pela tomografia de coerência óptica de domínio espectral.

Spontaneous outer retinal layer recovery in a case of hypertensive choroidopathy secondary to pre-eclampsia: a multimodal evaluation.

A 26-year-old woman presented at 28 weeks gestation with hypertensive choroidopathy associated with pre-eclampsia. Fundus photography, fundus autofluorescence, spectral-domain optical coherence tomography (SD-OCT), fluorescein angiography, and indocyanine green angiography were performed in both eyes in the immediate postoperative period. SD-OCT images were obtained before delivery and during a 3-month follow-up. Fundus autofluorescence exhibited patchy hyper- and hypoautofluorescent lesions; fluorescein and indocyanine green angiography revealed areas of choroidal ischemia; and SD-OCT showed disorganization of the outer retinal layers and disruption of the ellipsoid zone. After her blood pressure was stabilized, progressive recovery of the outer retinal layer was monitored on SD-OCT.

Vigilância entomológica de vetores de arbovírus na cidade de São Paulo: análise espaço temporal de criadouros, de acordo com fatores sazonais e socioeconômicos, no período de 2012 a 2016 / Entomological surveillance of arbovirus vectors in the city of São Paulo analysis of breeding sites, according to seasonal and socioeconomic factors, between 2012 and 2016

Introdução: Conhecer e monitorar fatores associados a transmissão de arboviroses são um grande desafio para os gestores de saúde pública e também uma necessidade para regiões onde há registro da circulação de arbovírus e a presença de vetores. Objetivos: Identificar os principais grupos de recipientes que podem se tornar ou são criadouros de mosquitos vetores no Município de São Paulo e verificar se os mesmos sofrem influências de fatores sazonais e socioeconômicos em sua distribuição. Método: Foram levantados dados das inspeções realizadas nas atividades de vigilância e controle de Aedes aegypti no período de 2012-2016, por Supervisão Técnica de Saúde e para o município. Em seguida foram construídos grupos de recipientes para avaliar predominância nas diferentes condições de encontro (existente, com água, com larva). Os indicadores gerados foram utilizados para testar diferenças significativas das frequências entre as estações do ano e analisar correspondência entre incidência de casos dengue e predominância de grupos de criadouros. Posteriormente, foi realizada análise de agrupamento por fatores socioeconômicos para identificar diferenças na distribuição dos grupos de recipientes. Para as análises de variância foi utilizado o teste não paramétrico de Kruskal-Wallis. Resultados: O grupo de recipientes móveis foi o potencial criadouro mais frequente em todos os anos, seguido dos grupos planta e pneus. Móvel e planta foram os criadouros mais frequentes para o município. O grupo móveis, apesar de numeroso, não foi o mais colonizado. Por outro lado, os depósitos para armazenamento de água tiveram baixa frequência, porém alta proporção de colonização. A sazonalidade influenciou na distribuição de recipientes com água e com larva, sendo que as maiores frequências ocorreram no verão e outono. A proporção de imóveis tendo recipientes com água aumentou de acordo com o tempo, principalmente em 2015. O grupo que teve maior incremento do índice nesse período foi o de depósito não ligado à rede. Foram gerados cinco grupos de STS por condições socioeconômicas, nos quais houve diferenças no padrão de distribuição de potenciais criadouros e criadouros. Regiões com melhor condição socioeconômica apresentaram menor frequência dos indicadores, mas não necessariamente menor risco de transmissão de dengue no período avaliado. Conclusões: Os principais grupos de criadouros mais frequentes no Município de São Paulo dentro do período analisado foram os de menor tamanho, principalmente os do tipo móvel. Reservatórios de maior tamanho e destinados ao armazenamento de água demonstraram importância na proporção de colonização e tiveram sua frequência aumentada no período da crise hídrica do abastecimento. Fatores sazonais e condições socioeconômicas influenciaram a distribuição de criadouros em São Paulo. Para melhor direcionamento das ações de prevenção e controle de vetores, analises sistemáticas e continuas por regiões devem ser realizadas

Introduction: Knowing and controlling factors associated with arbovirus transmission is a major challenge for public health managers, but it is necessary for regions where arbovirus circulation and the presence of vectors are present. Objectives: To identify the main groups of containers that can become or are breeding sites of mosquito vectors in the São Paulo Municipality and verify if their distribution are influenced by seasonal and socioeconomic factors. Method: Data were collected from the inspections carried out in the surveillance and control activities of Ae. aegypti in the period 2012-2016, by Health Technical Supervision and county. Groups of containers were then created to evaluate predominance in the different checked conditions (existing, with water, with larvae). The generated indicators were used to test significant differences of the frequencies between the seasons of the year and to analyze correspondence between incidence of dengue cases and predominance of breeding sites groups. After, a socioeconomic grouping analysis was performed to identify differences in the distribution of groups of recipients. The Kruskal-Wallis non-parametric test was used for the variance analysis. Results: The group of mobile containers was the most frequent potential breeding site in all the years, followed by container plants and tires groups. Mobile and plant were the most frequent breeding sites for the county. The mobile group, although numerous, was not the most colonized. On the other hand, deposits for water storage had a low frequency, but a high proportion of colonization. Seasonality influenced the distribution of water and larvae containers, with the highest frequencies occurring in summer and fall. The proportion of buildings having containers with water increased over time, especially in 2015. The group that had the largest increase in the index in this period was the deposit not connected to the water supply network. Five Health Technical Supervision groups were generated by socioeconomic conditions, in which there were differences in the pattern of distribution of potential breeding sites and breeding sites for mosquitoes. Regions with better socioeconomic status had lower frequency of indicators, but not necessarily a lower risk of dengue transmission in the period evaluated. Conclusions: The main breeding sites groups most frequent in the city of São Paulo during the analyzed period were the smaller groups, mainly those of the mobile type. Larger reservoirs destined to water storage showed importance in the proportion of colonization and had their frequency increased in the period of the water supply crisis. Seasonal factors and socioeconomic conditions influenced the distribution of breeding sites in. To better target vector prevention and control actions, systematic and continuous analyzes by regions should be performed

Immunomodulation in the canine endometrium by uteropathogenic Escherichia coli.

This study was designed to evaluate the role of E. coli α-hemolysin (HlyA) in the pathogenesis of canine pyometra, and on the immune response of canine endometrial epithelial and stromal cells. In Experiment 1, the clinical, hematological, biochemical and uterine histological characteristics of ß-hemolytic and non-hemolytic E. coli pyometra bitches were compared. More (p < 0.05) metritis cases were observed in ß-hemolytic E. coli pyometra uteri than in non-hemolytic E. coli pyometra uteri. ß-hemolytic E. coli pyometra endometria had higher gene transcription of IL-1ß and IL-8 and lower gene transcription of IL-6 than non-hemolytic E. coli pyometra endometria (p < 0.01). In Experiment 2, the immune response of endometrial epithelial and stromal cells, to hemolytic (Pyo18) and non-hemolytic E. coli strains (Pyo18 with deleted hlya-Pyo18ΔhlyA- and Pyo14) were compared. Following 4 h of incubation, Pyo18 decreased epithelial cell numbers to 54% (p < 0.001), and induced death of all stromal cells (p < 0.0001), whereas Pyo18ΔhlyA and Pyo14 had no effect on cell numbers. Compared to Pyo18ΔhlyA and Pyo14, respectively, Pyo18 induced a lower transcription level of IL-1ß (0.99 vs 152.0 vs 50.9 fold increase, p < 0.001), TNFα (3.2 vs 49.9 vs 12.9 fold increase, p < 0.05) and IL-10 (0.4 vs 3.6 vs 2.6 fold increase, p < 0.001) in stromal cells, after 1 h of incubation. This may be seen as an attempt of hemolytic E. coli to delay the activation of the immune response. In conclusion, endometrial epithelial and stromal cell damage induced by HlyA is a potential relevant step of E. coli virulence in the pathogenesis of pyometra.

Two SmDLC antigens as potential vaccines against schistosomiasis.

The Schistosoma mansoni transcriptome revealed new members of the dynein light chain family (DLC/LC8). The antigenicity and immunogenicity of these proteins, and their potential as vaccine candidates were investigated. Two DLC genes (DLC12_JI392413.1 and DLC13_JI387686.1) were cloned and the recombinant proteins produced in E. coli. The immunization of mice with the rDLCs, using alhydrogel as adjuvant, resulted in high titers of antibodies, indicated that these proteins are highly immunogenic. The anti-DLCs antibodies presented cross reactivity with both recombinant antigens and also recognized proteins from S. mansoni adult worm extracts. The DLC12 and DLC13 immunized animals were challenged by infection with cercariae and a protective profile was observed in three different assays, with a significant decreased in worm burden, of 43% and 51% respectively, when compared to the non-vaccinated group. The granulomas formation due to egg retention in the hepatic tissues was evaluated 45 days after infection. Smaller granulomas were observed in the liver of DLC immunized animals, up to 70% reduction in comparison to the granulomas size in the non-vaccinated animals. Fifty-five days after infection, the average size of the hepatic granulomas was still 25-35% smaller in the DLCs vaccinated groups. The interference of DLC immunization on the hepatic granuloma formation may reflect the lower worm burden and consequent decrease on the number of eggs retained in the liver, resulting in lower pro-inflammatory level in the tissue. The protective effect of DLCs immunization, decreasing the worm burden and delaying the rate of granuloma formation, suggests that these antigens should be further studied as potential vaccine candidates.

Lysophosphatidic acid signaling in late cleavage and blastocyst stage bovine embryos.

Lysophosphatidic acid (LPA) is a known cell signaling lipid mediator in reproductive tissues. In the cow, LPA is involved in luteal and early pregnancy maintenance. Here, we evaluated the presence and role of LPA in bovine early embryonic development. In relevant aspects, bovine embryos reflect more closely the scenario occurring in human embryos than the mouse model. Transcription of mRNA and protein expression of enzymes involved in LPA synthesis (ATX and cPLA2) and of LPA receptors (LPAR1-4) were detected in Days 5 and 8 in vitro produced embryos. Embryonic LPA production into culture medium was also detected at both stages of development. Supplementation of culture medium with LPA (10(-5) M) between Days 2 and 8 had no effect on embryo yield and quality and on blastocyst relative mRNA abundance of genes involved in prostaglandin synthesis (PTGS2, PGES, and PGFS) and steroidogenesis (3ßHSD). However, LPA treatment affected transcription levels of embryo quality markers, decreasing BAX (apoptotic) and increasing BCL2 (antiapoptotic) and IGF2R (growth marker) gene transcription levels. Blastocyst transcription of OCT4 (pluripotency marker) was not affected by LPA stimulation. In conclusion, LPA is an early bovine embryonic autocrine/paracrine signaling mediator, and LPA action may be relevant in early embryo-maternal interactions leading to embryonic survival.

The effect of lysophosphatidic acid during in vitro maturation of bovine oocytes: embryonic development and mRNA abundances of genes involved in apoptosis and oocyte competence.

In the present study we examined whether LPA can be synthesized and act during in vitro maturation of bovine cumulus oocyte complexes (COCs). We found transcription of genes coding for enzymes of LPA synthesis pathway (ATX and PLA2) and of LPA receptors (LPAR 1-4) in bovine oocytes and cumulus cells, following in vitro maturation. COCs were matured in vitro in presence or absence of LPA (10(-5) M) for 24 h. Supplementation of maturation medium with LPA increased mRNA abundance of FST and GDF9 in oocytes and decreased mRNA abundance of CTSs in cumulus cells. Additionally, oocytes stimulated with LPA had higher transcription levels of BCL2 and lower transcription levels of BAX resulting in the significantly lower BAX/BCL2 ratio. Blastocyst rates on day 7 were similar in the control and the LPA-stimulated COCs. Our study demonstrates for the first time that bovine COCs are a potential source and target of LPA action. We postulate that LPA exerts an autocrine and/or paracrine signaling, through several LPARs, between the oocyte and cumulus cells. LPA supplementation of maturation medium improves COC quality, and although this was not translated into an enhanced in vitro development until the blastocyst stage, improved oocyte competence may be relevant for subsequent in vivo survival.

Embryo-luteal cells co-culture: an in vitro model to evaluate steroidogenic and prostanoid bovine early embryo-maternal interactions.

The role of progesterone (P(4)) and prostaglandins (PGs) in bovine early embryonic development and embryo-maternal crosstalk is almost unknown. Here, the in vitro steroidogenic (P(4)) and prostanoid (PGE(2) and PGF(2α)) interactions between bovine embryos and luteal cells (LC) were evaluated. In two experiments, embryos (n = 1.900) were either co-cultured with LC or cultured alone, from days 2 to 7 (day 0 = in vitro insemination). LC were also cultured alone, and medium was used as a control, all groups being cultured either with or without oil overlay of culture medium. Oil overlay of culture medium significantly decreased the amount of P(4), but not of PGE(2) and PGF(2α) measured in culture medium. Embryos and LC had transcripts of genes coding for enzymes of the PGs (PTGS2, PGES, and PGFS) and P(4) (StAR, P450scc, and 3ß-HSD) synthesis pathways, and produced P(4), PGF(2α), and PGE(2) into culture medium. Co-culture with LC exerted an embryotrophic effect, significantly increasing blastocyst yield and quality. This indicates a possible direct effect of LC in early embryo development. Embryos did not exert a luteotrophic effect upon LC. This may indicate that early embryos (until day 7) probably do not exert influence in LC main function. It is suggested that production of P(4), PGE(2), and PGF(2α) by early embryos may be associated to autocrine signaling leading to events in development and to paracrine signaling in the endometrium leading to local uterine receptivity.

Fallopian tube origin of supposed ovarian high-grade serous carcinomas.

INTRODUCTION: Serous carcinomas are the most frequent histologic type of ovarian and peritoneal cancers, and can also be detected in the endometrium and fallopian tubes. Serous carcinomas are usually high-grade neoplasms when diagnosed, yet the identification of an associated precursor lesion remains challenging. Pathological examination of specimens obtained from prophylactic bilateral salpingo-oophorectomies that were performed for patients harboring BRCA1/2 mutations suggests that high-grade serous carcinomas may arise in the fallopian tubes rather than in the ovaries. OBJECTIVE: To investigate the presence and extent of fallopian tube involvement in cases of serous pelvic carcinomas. METHODS: Thirty-four cases of serous pelvic carcinoma with clinical presentations suggesting an ovarian origin were analyzed retrospectively. Histologic samples of fallopian tube tissues were available for these cases and were analyzed. Probable primary site, type of tubal involvement, tissues involved in the neoplasia and vascular involvement were evaluated. RESULTS: Fallopian tube involvement was observed in 24/34 (70.6%) cases. In 4 (11.8%) of these cases, an intraepithelial neoplasia was present, and therefore these cases were hypothesized to be primary from fallopian tubes. For an additional 7/34 (20.6%) cases, a fallopian tube origin was considered a possible primary. CONCLUSIONS: Fallopian tubes can be the primary site for a subset of pelvic high-grade serous carcinomas.

Lack of circadian variation of pulse wave velocity measurements in healthy volunteers.

Arterial stiffness is an independent marker of cardiovascular events. Pulse wave velocity (PWV) is a validated method to detect arterial stiffness that can be influenced by several factors including age and blood pressure. However, it is not clear whether PWV could be influenced by circadian variations. In the present study, the authors measured blood pressure and carotid-femoral PWV measurements in 15 young healthy volunteers in 4 distinct periods: 8 am, noon, 4 pm, and 8 pm. No significant variations of systolic (P=.92), mean (P=.77), and diastolic (P=.66) blood pressure among 8 am (113±15, 84±8, 69±6 mm Hg), noon (114±13, 83±8, 68±6 mm Hg), 4 pm (114±13, 85±8, 70±7 mm Hg), and 8 pm (113±7, 83±10, 68±7 mm Hg), respectively, were observed. Similarly, carotid-femoral PWV did not change among the periods (8 am: 7.6 ± 1.4 m/s, noon: 7.4±1.1 m/s, 4 pm: 7.6±1.0 m/s, 8 pm, 7.6±1.3 m/s; P=.85). Considering all measurements, mean blood pressure significantly correlated with PWV (r=.31; P=.016). In young healthy volunteers, there is no significant circadian variation of carotid-femoral PWV. These findings support the concept that it does not appear mandatory to perform PWV measurements at exactly the same period of the day.

The effects of continuous positive airway pressure on prehypertension and masked hypertension in men with severe obstructive sleep apnea.

Obstructive sleep apnea and hypertension are common conditions that frequently coexist. Continuous positive airway pressure (CPAP) reduces blood pressure in patients with obstructive sleep apnea and sustained hypertension. However, the impact of CPAP on patients with obstructive sleep apnea and prehypertension and masked hypertension, conditions associated with increased cardiovascular risk, is unknown. Thirty-six male patients (age, 43 ± 7 years; body mass index, 28.8 ± 3.0 kg/m(2)) with untreated severe obstructive sleep apnea (apnea-hypopnea index, 56 ± 22 events/hr on polysomnography) with diagnostic criteria for prehypertension and/or masked hypertension, based on office and 24-hour ambulatory blood pressure monitoring, respectively, were studied. The patients randomized to no treatment (control; n=18) or CPAP (n=18) for 3 months had similar frequency of prehypertension and masked hypertension at study entry. There were no significant changes in blood pressure in patients randomized to the control group. In contrast, patients randomized to CPAP presented significant reduction in office systolic (from 126 ± 5 to 121 ± 7 mm Hg; P=0.001) and a trend for diastolic blood pressure (from 75 ±7 to 73 ± 8 mm Hg; P=0.08) as well as a significant decrease in daytime and nighttime systolic and diastolic blood pressure (P<0.05 for each comparison). There was a significant reduction in the frequency of prehypertension (from 94% to 55%; P=0.02) and masked hypertension (from 39% to 5%; P=0.04) only in the CPAP group. In conclusion, effective CPAP therapy promotes significant reduction in the frequency of prehypertension and masked hypertension by promoting significant blood pressure reductions in patients with severe obstructive sleep apnea.

Fallopian tube origin of supposed ovarian high-grade serous carcinomas

INTRODUCTION: Serous carcinomas are the most frequent histologic type of ovarian and peritoneal cancers, and can also be detected in the endometrium and fallopian tubes. Serous carcinomas are usually high-grade neoplasms when diagnosed, yet the identification of an associated precursor lesion remains challenging. Pathological examination of specimens obtained from prophylactic bilateral salpingo-oophorectomies that were performed for patients harboring BRCA1/2 mutations suggests that high-grade serous carcinomas may arise in the fallopian tubes rather than in the ovaries. OBJECTIVE: To investigate the presence and extent of fallopian tube involvement in cases of serous pelvic carcinomas. METHODS: Thirty-four cases of serous pelvic carcinoma with clinical presentations suggesting an ovarian origin were analyzed retrospectively. Histologic samples of fallopian tube tissues were available for these cases and were analyzed. Probable primary site, type of tubal involvement, tissues involved in the neoplasia and vascular involvement were evaluated. RESULTS: Fallopian tube involvement was observed in 24/34 (70.6 percent) cases. In 4 (11.8 percent) of these cases, an intraepithelial neoplasia was present, and therefore these cases were hypothesized to be primary from fallopian tubes. For an additional 7/34 (20.6 percent) cases, a fallopian tube origin was considered a possible primary. CONCLUSIONS: Fallopian tubes can be the primary site for a subset of pelvic high-grade serous carcinomas.

Obstructive sleep apnea, masked hypertension, and arterial stiffness in men.

BACKGROUND: Obstructive sleep apnea (OSA) is an established cause of hypertension. However, it is not clear whether the frequency of masked hypertension in patients with OSA and whether OSA have an independent role on arterial stiffness taking into account ambulatory blood pressure (BP) monitoring (ABPM). METHODS: We evaluated 61 male normotensive participants as determined by casual clinic BP level <140/90 mm Hg without clinical evidence of cardiovascular disease and on no medications (43 patients with moderate-to-severe OSA (apnea-hypopnea index (AHI) > or = 15 events/hour by polysomnography) and 18 age- and body mass index-matched controls without OSA (AHI <5 events/hour)). Pulse wave velocity (PWV), an index of arterial stiffness, and 24-h ABPM were performed in a blinded fashion. Masked hypertension was defined when abnormal daytime ABPM was > or = 135 or > or = 85 mm Hg. RESULTS: The AHI and lowest oxygen saturation were 2.6 +/- 1.6 and 90 +/- 2 vs. 52.8 +/- 21.0 events/hour and 75 +/- 10% for controls and OSA patients, respectively; P < 0.001. Compared with controls, patients with OSA had higher office systolic BP (113 +/- 9 vs. 118 +/- 10 mm Hg; P = 0.05) and a higher unadjusted proportion of masked hypertension (2 controls (11.1%) vs. 13 patients (30.2%); P < 0.05). PWV was 8.7 +/- 0.7, 9.4 +/- 1.0, and 10.6 +/- 1.1 m/s in the control, OSA without and with masked hypertension groups, respectively (P < 0.01 for each comparison). Multiple regression showed that systolic daytime ABPM and the lowest oxygen saturation were independently related to PWV (adjusted R2 = 0.34; P < 0.01). CONCLUSIONS: Patients with OSA presented a higher unadjusted rate of masked hypertension than matched controls. Lowest oxygen saturation has an independent association with arterial stiffness.