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To compare the expression profile of extracellular vesicle microRNAs (EV-miRNAs) derived from follicular fluid after a trigger with recombinant human chorionic gonadotropin (r-hCG) or with a gonadotropin-releasing hormone GnRH agonist (GnRH-a) for final oocyte maturation.A retrospective analysis of a prospective cohort. Women undergoing in vitro fertilization at a tertiary university-affiliated hospital were recruited between 2014 and 2016. EV-miRNAs were extracted from the follicular fluid of a single follicle, and their expression was assessed using TaqMan Open Array®. Genes regulated by EV-miRNAs were analyzed using miRWalk2.0 Targetscan database, DAVID Bioinformatics Resources, Kyoto-Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO).Eighty-two women were included in the r-hCG trigger group and 9 in the GnRH-a group. Of 754 EV-miRNAs screened, 135 were detected in at least 50% of the samples and expressed in both groups and were further analyzed. After adjusting for multiple testing, 41 EV-miRNAs whose expression levels significantly differed between the two trigger groups were identified. Bioinformatics analysis of the genes regulated by these EV-miRNAs showed distinct pathways between the two triggers, including TGF-beta signaling, cell cycle, and Wnt signaling pathways. Most of these pathways regulate cascades associated with apoptosis, embryo development, implantation, decidualization, and placental development.Trigger with GnRH-a or r-hCG leads to distinct EV-miRNAs expression profiles and to downstream biological effects in ovarian follicles. These findings may provide an insight for the increased apoptosis and the lower implantation rates following GnRH-a trigger vs. r-hCG in cases lacking intensive luteal phase support.

Study question Can sleep quality and psychological health of women undergoing In Vitro Fertilization (IVF) affect the chances of success of the procedure? Summary answer Psychological health doesn’t seem to affect IVF outcomes while sleep disturbances are common in women undergoing IVF and could influence the success of the procedure. What is known already IVF is saddled with a significant emotional burden causing anxiety, depression, and sleep disturbances. Baseline high levels of stress seem to be negatively associated with IVF success whereas studies evaluating the effect of anxiety and depression are discordant. Investigating sleep disorders in this setting is particularly attracting because of the significant association with anxiety and stress and because it is a simple information to collect. In addition, sleep disorders could negatively affect IVF success, as it does for natural conceptions, due to the well-established association between sleep and the hormonal system. Evidence on this issue is however scant and inconclusive. Study design, size, duration This is a single-center prospective cohort study designed to examine whether the sleep quality and psychological health of women undergoing IVF affect the chances of success of the procedure. Two hundred sixty-three women undergoing IVF between September to December 2019 were enrolled. Baseline characteristics and women’s sleep quality and psychological health were compared between women who did and did not have a live birth. Participants/materials, setting, methods Inclusion criteria were admission for oocyte pick-up. Women with neurologic or psychiatric disorders, obstructive sleep apnea syndrome, anatomical factors that can reduce embryo implantation and limited understanding of the Italian language were excluded. Women’s sleep quality and psychological health were assessed using the Pittsburgh Sleep Quality Index (PSQI), the Fertility Problem Inventory (FPI), and the Hospital Anxiety and Depression Scale (HADS). Main results and the role of chance Overall, 81 women had a live birth (31%). As expected, successful women were younger, and their ovarian reserve was more preserved. No significant group differences were detected for education, employment, parity, smoking status, body mass index and indication to IVF. FPI and HADS scores did not differ. Conversely, our findings revealed that sleep disturbances were associated with lower chances of live birth in IVF: a statistically significant difference emerged for the PSQI score, the median [interquartile range] in pregnant and non-pregnant women being 4 [3-5] and 5 [3-7], respectively (p = 0.004). The crude and adjusted OR of pregnancy in women with a PSQI >5 (indicating impaired sleep quality) was 0.46 (95%CI: 0.25-0.86, p = 0.02) and 0.50 (95%CI: 0.26-0.94, p = 0.03), respectively. Limitations, reasons for caution Data were collected during treatment, which did not allow to properly investigate the association between sleep quality, infertility, and IVF itself. Sleep quality was assessed using a self-reported sleep measure (although validated and extensively used by clinicians and researchers), which does not ensure the objectivity of observations. Wider implications of the findings The assessment of sleep quality throughout the treatment period should be included as an important part of multidisciplinary clinical management of women undergoing IVF. Sleep quality of candidate mothers should be promoted by the implementation of prevention programmes and targeted interventions. Trial registration number not applicable

Study question Can the UF-EVs transcriptome provide markers of implantation in women diagnosed with recurrent implantation failure (RIF)? Summary answer Enrichment in IL-10 and IL-17 pathway transcripts in UF-EVs of women with RIF might identify those at risk for negative outcome after euploid blastocyst transfer. What is known already Our group has recently found that transcriptomic changes occur in UF-EVs during the window of implantation and closely resemble those occurring in the endometrial tissue. Patients affected by RIF represent a debated and heterogeneous population of patients who currently undergo several investigations, empirical therapies and embryo transfer attempts, with inconclusive results. We investigated whether UF-EVs can provide markers to define subgroups of patients within the RIF population, to predict outcomes and tailor interventions. Study design, size, duration We herein present the first sub-analysis of our previous larger observational study on the human UF-EVs transcriptome (Giacomini et al., 2021). RIF was defined by > 3 failed embryo transfers (ETs) with high quality embryos and patients affected by RIF and undergoing their first PGT-A cycle were enrolled (n = 19). Fertile women undergoing PGT-A+PGT-M for monogenic diseases and achieving successful implantation were used as controls (n = 29). PGT-A was performed by Next Generation Sequencing. Participants/materials, setting, methods RNA-Seq was performed on UF-EVs collected on day LH + 7 of the cycle preceding that of the blastocyst transfer. Differential Gene Expression (DGE) analysis was performed between RIF patients failing implantation (Group A) versus RIF patients achieving implantation (Group B) and between Group A and fertile controls achieving implantation (Group C). Pre-ranked gene set enrichment (GSEA) with WebGestalt was used for pathway enrichment analysis, with Normalized Enrichment Score (NES) indicating the strength of the enrichment. Main results and the role of chance RIF patients had a mean (± SD) age of 36.6 (± 3.4) and 5.1 ± 2.1 (mean ± SD) previous failed ETs. DGE comparisons between UF-EVs of RIF women who failed implantation (n = 10) showed 135 up- and 32 down-regulated genes compared to Group B, and 258 up- and 44 down-regulated genes compared to Group C, with 63 genes showing consistent significantly different ‘expression’ values in both comparisons. UF-EVs of Group A were enriched in transcripts belonging to the IL-17 pathway compared to both groups (Group A vs B, NES=2.2044, FDR Limitations, reasons for caution As the main limitation of our study, we collected UFs in the cycle immediately preceding (and not corresponding to) the euploid ET. This approach was considered the most appropriate in relation to the current paucity of data regarding the safety of UF aspiration in the same cycle of an ET. Wider implications of the findings UF-EVs represent a source of transcriptomic markers related to the endometrial immune profile in RIF patients. Enrichment in the IL-17 and IL-10 pathways in this population could define the subgroup who is at risk for implantation failure even after euploid ET and who could potentially benefit from tailored immunotherapies. Trial registration number not applicable

Objective: Exposure to airborne pollutants during pregnancy appears to be associated with uterine growth restriction and adverse neonatal outcome. Proprotein convertase subtilisin/kexin type (PCSK9), the key modulator of low-density lipoprotein (LDL) metabolism, increases following particulate matter (PM10) exposure. Because maternal cholesterol is required for fetal growth, PCSK9 levels could be used to evaluate the potential impact of airborne pollutants on fetal growth. Design: A cohort of 134 healthy women during early pregnancy (11–12 weeks of gestational age) was studied. Results: A significant association between circulating PCSK9 levels and three tested air pollutants (PM10, PM2.5, nitric oxide (NO2)) was found. Of importance, gestational age at birth was reduced by approximately 1 week for each 100 ng/mL rise in circulating PCSK9 levels, an effect that became more significant at the highest quartile of PM2.5 (with a 1.8 week advance in delivery date for every 100 ng/mL rise in circulating PCSK9; p for interaction = 0.026). This finding was supported by an elevation of the odds ratio for urgent cesarean delivery for each 100 ng/mL rise in PCSK9 (2.99, 95% CI, 1.22–6.57), similar trends being obtained for PM10 and NO2. Conclusions: The association between exposure to air pollutants during pregnancy and elevation in PCSK9 advances our understanding of the unforeseen influences of environmental exposure in terms of pregnancy associated disorders.

BackgroundAccumulating evidence suggests that poor mental health is one of the most common comorbidities of both rheumatoid arthritis (RA) and osteoarthritis (OA) [1]. Even if underpinning RA and OA are different genetic, structural, mechanical, and immunologic pathways involved in their pathogenesis, poor mental health, and joint involvement are intertwined and negatively affect their mutual course by contributing to global disability. Thus, new insights into mechanisms that link these disorders are needed to identify new actionable biomarkers to drive more personalized therapeutic strategies. Amidst potential mediators, extracellular vesicles (EVs) play a central role in terms of communication between cells, they cross the blood-brain barrier and based on their cargos can affect the recipient cell function [2].ObjectivesTo isolate EVs from synovial fluid (SF) in RA and OA patients and to evaluate if and how these EVs can alter in vitro synaptic transmission of murine hippocampal neurons.MethodsIn this cross-sectional pilot study, consecutive adult RA and primary OA who were referred to the Rheumatology Unit for aspiration of joint effusion were enrolled. Demographic and clinical variables and mental health rating scales were collected. Discarded SF were collected and EVs were isolated and analyzed by Malvern NanoSight NS300 system to obtain information on their number and size. Afterwards, DIV14 cultured wild-type hippocampal neurons were exposed for two hours to OA- and RA-EVs at low and high concentration EVs. Thus, miniature excitatory and inhibitory postsynaptic currents (mEPSCs and mIPSCs), which reflects glutamatergic and GABA-ergic activity respectively, were examined by exploiting patch-clamp recordings in the whole-cell configuration. Frequency and amplitude were analyzed to evaluate potential changes at the presynaptic or postsynaptic compartment. Mann Whitney test was used to compare two different samples.ResultsEight RA patients (7 female, mean age 57 yrs), and 5 primary OA (4 female, mean age 60 yrs) were recruited for SF aspiration. The mean VAS pain was 7.25 in RA and 6.5 in OA. No statistically significant differences were found between the two groups in mean rating scale scores although patients affected by RA had more severe depressive symptoms (Montgomery Asberg Depression Rating Scale -MADRS- means scores: 16.57) with respect OA group (MADRS mean scores: 10). The Nanoparticle tracking analysis showed that RA-EVs were significantly more in number compared to OA-EVs (Figure 1 A), mimicking more inflammation, while no significant difference in size was observed. Analysis of miniature events revealed the occurrence of two different changes. High concentration of OA-EVs has led to an increased amplitude of excitatory events, meaning an increased susceptibility of neurons to glutamate in the post-synaptic compartment (Figure 1 B). Whereas low concentration of RA-EVs has led to a decreased frequency of inhibitory events, which reflects a reduced function of GABA-ergic synapse in the pre-synaptic compartment (Figure 1 C).Figure 1.ConclusionOur results suggest that SF-derived EVs from OA and RA patients lead to different specific changes of neurotransmission, with different concentration needed to alter neuronal spontaneous activity in post-synaptic and pre-synaptic compartment, respectively. EVs may provide insight into the pathogenesis of joint-brain communication in RA and OA, unraveling specific pathways thus allowing targeted therapies for neuropsychiatric involvement.References[1]Lancet 2017;390(10100): 1211–1259[2]FASEB Bioadv 2021;3(9):665-675Disclosure of InterestsLavinia A. Coletto: None declared, Francesca Ingegnoli: None declared, Clara Cambria: None declared, Laura Cantone: None declared, Orazio De Lucia: None declared, Roberto Caporali Speakers bureau: Abbvie, Amgen, BMS, Celltrion, Galapagos, Lilly, Pfizer, Fresenius-Kabi, MSD, UCB, Roche,Janssen, Novartis, Sandoz, Consultant of: Abbvie, Amgen, BMS, Celltrion, Galapagos, Lilly, Pfizer, MSD, UCB, Janssen, Novartis, Sandoz, Valentina Bollati: None declared, Massimiliano Buoli: None declared, Flavia Antonucci: None declared.

Exposure to airborne pollutants during pregnancy appears to be associated with uterine growth restriction and adverse neonatal outcome. Proprotein convertase subtilisin/kexin type (PCSK9), the key modulator of low-density lipoprotein (LDL) metabolism, increases following particulate matter (PMA cohort of 134 healthy women during early pregnancy (11-12 weeks of gestational age) was studied.A significant association between circulating PCSK9 levels and three tested air pollutants (PMThe association between exposure to air pollutants during pregnancy and elevation in PCSK9 advances our understanding of the unforeseen influences of environmental exposure in terms of pregnancy associated disorders.

Additional file 1: Table S1.Antidepressant treatments (n=37). Figure S1. Slope coefficients from univariate linear regression models evaluating the association between levels of PCSK9 and characteristics of subjects.

Not significant data is presented as the estimate (Ă ) and their respective p-value (p). Disease activity is presented as â annualized relapse rateâ (ARR). The presence and amount of oligoclonal bands was measured in cerebrospinal fluid (CSF). (DOCX 18 kb)

CpG specific odds ratios were calculated between MS (n = 51) patients and Healthy controls (n = 137 for LINE-1 and SAT-Îą; n = 135 for Alu). Methylation was assessed in 3 CpG sites for Alu and SAT-Îą and 4 CpG sites in LINE-1. Estimates are presented as odds ratios, adjusted for age, gender and smoking status, and were calculated using the multivariate logistic regression analysis. (DOCX 564 kb)

Characteristics of Multiple Sclerosis (MS) patients and healthy controls for each methylation marker. Methylation is subdivided as â meanâ (i.e. average of the separate positions) and the individual positions of the markers. (DOCX 66 kb)

Airborne exposure to particulate matter with diameter 10 mcM (PM10) has been linked to an increased risk of thromboembolic events, but the mechanisms are not completely understood. The aim of this study was to evaluate the effect of PM10 phagocytosis on the release of procoagulant molecules in human differentiating macrophages, and that of PM10 inhalation in an experimental model in rats. Human monocytes were separated from the peripheral blood by the lymphoprep method, differentiated in vitro and treated with standard PM10 or vehicle. Sprague-Dawley rats were instilled intratracheally with PM10 or vehicle alone. The outcome was expression of proinflammatory genes and of tissue factor (TF). In human differentiating macrophages, PM10 exposure upregulated inflammatory genes, but most consistently induced TF mRNA and protein levels, but not TF protein inhibitor, resulting in increased TF membrane expression and a procoagulant phenotype. Differentiation towards the anti-inflammatory M2 phenotype inhibited PM10 -mediated TF expression. TF induction required phagocytosis of PM10 , whereas phagocytosis of inert particles was less effective. PM10 phagocytosis was associated with a gene expression profile consistent with intracellular retention of iron, inducing oxidative stress. Both PM10 and iron activated the stress kinases ERK1/2 pathway, involved in the induction of TF expression. In rats, alveolar exposure to PM10 was associated with pulmonary recruitment of inflammatory cells and resulted in local, but not systemic, induction of TF expression, which was sufficient to increase circulating TF levels. In conclusion, TF induction by differentiating lung macrophages, activated following phagocytosis, contributes to the increased risk of thromboembolic complications associated with PM10 exposure.