Preeclampsia (PE) is a hypertensive complication of pregnancy that affects 2–8% of women worldwide and is one of the leading causes of maternal deaths and premature birth. PE can occur early in pregnancy (<34 weeks gestation) or late in pregnancy (>34 weeks gestation). Whilst the placenta is clearly implicated in early onset PE (EOPE), late onset PE (LOPE) is less clear with some believing the disease is entirely maternal whilst others believe that there is an interplay between maternal systems and the placenta. In both types of PE, the syncytiotrophoblast (STB), the layer of the placenta in direct contact with maternal blood, is stressed. In EOPE, the STB is oxidatively stressed in early pregnancy (leading to PE later in gestation- the two-stage model) whilst in LOPE the STB is stressed because of villous overcrowding and senescence later in pregnancy. It is this stress that perturbs maternal systems leading to the clinical manifestations of PE. Whilst some of the molecular species driving this stress have been identified, none completely explain the multisystem nature of PE. Syncytiotrophoblast membrane vesicles (STB-EVs) are a potential contributor to this multisystem disorder. STB-EVs are released into the maternal circulation in increasing amounts with advancing gestational age, and this release is further exacerbated with stress. There are good in vitro evidence that STB-EVs are taken up by macrophages and liver cells with additional evidence supporting endothelial cell uptake. STB-EV targeting remains in the early stages of discovery. In this review, we highlight the role of STB-EVs in PE. In relation to current research, we discuss different protocols for ex vivo isolation of STB-EVs, as well as specific issues involving tissue preparation, isolation (some of which may be unique to STB-EVs), and methods for their analysis. We suggest potential solutions for these challenges.

BACKGROUND: Preeclampsia is a major cause of maternal and perinatal morbidity and mortality worldwide. Identifying women with high risk of developing preeclampsia in early pregnancy remains challenging. Extracellular vesicles released from the placenta offer an attractive biomarker but have been elusive to quantify. METHODS: Here, we tested ExoCounter, a novel device that immunophenotypes size-selected small extracellular vesicles <160 nm, for its ability to perform qualitative and quantitative placental small extracellular vesicles (psEV) analysis. To investigate disease-specific and gestational age-specific changes, we analyzed psEV counts in maternal plasma samples taken at each of the 3 trimesters from women who had (1) normal pregnancy (n=3); (2) women who developed early-onset preeclampsia ([EOPE], n=3); and (3) women who developed late-onset preeclampsia (n=4) using 3 antibody pairs, CD10-placental alkaline phosphatase (PLAP), CD10-CD63, and CD63-PLAP. We further validated the findings in first-trimester serum samples among normal pregnancy (n=9), women who developed EOPE (n=7), and women who developed late-onset preeclampsia (n=8). RESULTS: We confirmed that CD63 was the major tetraspanin molecule coexpressed with PLAP—a known placental extracellular vesicles marker on psEV. Higher psEV counts for all 3 antibody pairs were detected in the plasma of women who developed EOPE than the other 2 groups in the first trimester, which persisted through the second and third trimesters. Significantly higher CD10-PLAP ( P <0.01) and CD63-PLAP ( P <0.01) psEV counts were validated in the serum of the first trimester of women who developed EOPE compared with normal pregnancy. CONCLUSIONS: Application of the ExoCounter assay developed here could identify patients at risk of developing EOPE in the first trimester, thereby providing a window of opportunity for early intervention.

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Background: Brassica oleracea L. occupies an important position in the annual production of vegetables. But during winter Brassica oleracea L. often suffers from low temperatures and even sub-zero temperatures. Through transcriptome analysis and identification, the pathways involved in cold tolerance of Brassica oleracea L. were analyzed and candidate genes related to cold tolerance of Brassica oleracea L. were identified.Results: Under low temperature stress, a large number of significantly different genes were found in Zhonggan1229 (ZG, low temperature tolerance) and Yingchun (YN, low temperature sensitive). There were 3902 significantly up-regulated genes and 5309 significantly down-regulated genes in ZG, and 4253 significantly up-regulated genes and 5938 significantly down-regulated genes in YN. Among them, 1844 different genes are the specific different genes in ZG and 6089 genes are the common different genes to response the low temperature stress. By annotating the specific different genes in ZG, 26 of the top 30 enriched GO terms belonged to biological processes, 4 terms belonged to molecular functions. By annotating the common different genes, 23 GO terms belonged to biological processes, 1 GO term belonged to molecular functions, and 6 GO terms belonged to cellular components. Circadian rhythms of plants and Plant hormone signal transduction were not only significantly enriched in the two analyzed genes, but also the effects of low temperature stress were most significant. Among the unique different genes in ZG, 154 genes were annotated into transcription factor families, and 79 genes were up-regulated and 75 genes were down-regulated, the encoding of MYB-related proteins was the largest group. Among the different genes shared by the two varieties, 516 genes were annotated into corresponding transcription factor families, 211 genes were up-regulated and 296 genes were down-regulated, however, there were 4 genes that were up-regulated in ZG but down-regulated in YN, and 5 genes that were down-regulated in ZG but up-regulated in YN, the largest group was the protein encoding ERF.Conclusions: The results identified important genes, pathways, and transcription factors that respond to low temperature stress, provided cold tolerance gene resources for the subsequent cold tolerance breeding research of Brassica oleracea L..