Cervicovaginal fluid proteomic analysis to identify potential biomarkers for preterm birth.

BACKGROUND: Spontaneous preterm birth is a leading cause of neonatal morbidity and mortality. Early identification of at-risk women by reliable screening tests could reduce the spontaneous preterm birth rate, but conventional methods such as obstetrical history and maternal cervical length screening identify only a minority of spontaneous preterm birth cases. Cervicovaginal fluid might prove to be a useful, readily available biological fluid for identifying spontaneous preterm birth biomarkers. OBJECTIVE: The objective of the study was to identify cervicovaginal fluid biomarkers of early spontaneous preterm birth in a high-risk cohort of pregnant women with a history of spontaneous preterm birth using targeted and shotgun proteomic analyses. STUDY DESIGN: A nested case control study (cases were spontaneous preterm birth <34 weeks in the current pregnancy; controls were spontaneous labor and delivery at 39-41 weeks) was performed using cervicovaginal fluid samples collected at 3 study visits (100/7 to 186/7 weeks, 190/7 to 236/7 weeks, and 280/7 to 316/7 weeks). All participants had a history of at least 1 prior spontaneous preterm birth. Targeted proteomic analysis was performed using a stable isotope-labeled proteome derived from endocervical and vaginal mucosal cells. This served as a standard to quantitate candidate protein levels in individual cervicovaginal fluid samples from the second and third study visits using liquid chromatography-multiple reaction monitoring mass spectrometry. The ratio of endogenous peptide area/stable isotope-labeled proteome-derived peptide area was used to measure levels of 42 peptides in 22 proteins. To maximize biomarker discovery in the cervicovaginal fluid samples, shotgun proteomic analysis also was performed utilizing liquid chromatography and ion trap mass spectrometry. A validation study was performed in second-trimester cervicovaginal fluid samples from an independent study group (12 spontaneous preterm birth cases, 19 term delivery controls) using enzyme-linked immunosorbent assay for 5 proteins expressed at higher levels in spontaneous preterm birth cases compared with controls in targeted or shotgun proteomic analyses. RESULTS: For targeted proteomics, cervicovaginal fluid samples from 33 cases and 32 controls at 190/7 to 236/7 weeks and 16 cases and 14 controls at 280/7 to 316/7 weeks from the same pregnancies were analyzed. When samples were compared between cases and controls, the relative abundance of 5 proteins was greater (P = .02-.05) in cases at both visits, while the relative abundance of 1 protein was lower (P = .03) in cases at both visits. For shotgun proteomics analyses, cervicovaginal fluid samples were pooled for 9 spontaneous preterm birth cases and 9 term delivery controls at each study visit. Shotgun proteomics yielded 28 proteins that were detected at levels >2 times higher and 1 protein that was detected at a level <0.5 times lower in spontaneous preterm birth cases compared with controls at all 3 study visits. Validation enzyme-linked immunosorbent assay for 5 proteins that were detected at higher levels in cervicovaginal fluid samples from spontaneous preterm birth cases compared with term delivery controls in proteomics analyses did not demonstrate statistically significant differences between spontaneous preterm birth cases and controls. CONCLUSIONS: Potential biomarkers of spontaneous preterm birth were identified by targeted and shotgun proteomics analyses in cervicovaginal fluid samples from high-risk, asymptomatic women. Many of the proteins detected at higher levels in cervicovaginal fluid samples from spontaneous preterm birth cases are extracellular matrix proteins and/or regulate cell membrane physiology. These proteins have substantial biological interest, but validation enzyme-linked immunosorbent assay for 5 of these proteins did not yield clinically useful biomarkers for spontaneous preterm birth.

Predicting persistent impaired glucose tolerance in patients with gestational diabetes: The role of high sensitivity CRP and adiponectin.

BACKGROUND: To evaluate whether maternal serum adiponectin and high-sensitivity C-reactive protein (hsCRP) levels at the time of gestational diabetes mellitus (GDM) diagnosis are associated with persistent glucose intolerance in GDM women at 6 to 12 weeks postpartum. METHODS: This is a secondary analysis of prospective randomized trial of GDM women enrolled in a behaviour education programme. Women with a GDM diagnosis ≥20 weeks were included. At the time of randomization, serum adiponectin and hsCRP levels were drawn. After delivery, women underwent a 2-hour 75-g oral glucose tolerance test at 6 to 12 weeks postpartum. Persistent impaired glucose tolerance (P-IGT) was defined as impaired fasting glucose, impaired glucose tolerance, or a diagnosis of type 2 diabetes mellitus. Regression models and receiver operator curves were used to evaluate the association between midpregnancy adiponectin and hsCRP and persistent impaired glucose tolerance. RESULTS: Of 100 women in the trial, 63 completed postpartum glucose testing. Twenty (31.7%) of the women had P-IGT. Median hsCRP levels were higher at randomization (22-34 wk) in women with persistent impaired glucose tolerance compared with women with normal glucose tolerance (5.1 vs 3.8, P = .01). After adjustment for the original study intervention, the association between hsCRP and P-IGT persisted (odds ratio, 3.45; 95% confidence interval, 1.34-8.92; P = .01) and had good diagnostic performance with an area under the curve of 0.73. There was no difference in median adiponectin levels between groups (44.8 vs 52.0, P = .57) or in odds of P-IGT (odds ratio, 0.81; 95% confidence interval, 0.33-1.99; P = .65), and area under the curve = 0.54. CONCLUSIONS: Midpregnancy high sensitivity CRP is a potential predictor of persistent impaired glucose tolerance diagnosed on the postpartum 2-hour 75-g oral glucose tolerance test in GDM women in the immediate postpartum period. Further investigation is needed in a larger population of women prior to using specific cut-offs for diagnostic purposes. High-sensitivity C-reactive protein levels in the immediate postpartum period should be seen as an adjunct, not a replacement, for the standard long-term screening of women with a history of a GDM pregnancy.

A Case of Genital Self-Mutilation in a Female-Symptom Choice and Meaning.

Self-mutilation is understood as any willful gesture or alteration of the body tissue without a suicidal intent. The most common self-mutilating gestures are, to a great extent, those that are superficial or moderate, such as cuts, burns, or bites. The most severe, as is the case of genital self-mutilation (GSM), are extremely rare and, in most cases, observed in patients suffering from psychosis. Furthermore, they are mostly reported from a surgical standpoint. Here, we report the case of a 20-year-old female patient who resorted to the emergency department after having amputated her clitoris with surgical scissors. This dramatic gesture, coupled with the patient's narrative, prompted for differential diagnosis between a psychotic syndrome and a severe personality disorder. We propose that, despite the magnitude of the self-harm, it is possible to conceptualize this GSM within a disturbed personality with significant sexuality issues and, therefore, this case report aims to broaden the limits that have been associated with the self-mutilating gestures in borderline personality disorder.

First-trimester placental ultrasound and maternal serum markers as predictors of small-for-gestational-age infants.

OBJECTIVE: The objective of the study was to combine early, direct assessment of the placenta with indirect markers of placental development to identify pregnancies at greatest risk of delivering small-for-gestational age infants (SGA10). STUDY DESIGN: We prospectively collected 3-dimensional ultrasound volume sets, uterine artery pulsatility index, and maternal serum of singleton pregnancies at 11-14 weeks. Placental volume (PV), quotient (placental quotient [PQ] = PV/gestational age), mean placental diameter (MPD) and chorionic diameters, and the placental morphology index (PMI = MPD/PQ and adjusts the lateral placental dimensions for quotient) were measured offline. Maternal serum was assayed for placental growth factor and placental protein-13. These variables were evaluated as predictors of SGA10. RESULTS: Of the 578 pregnancies included in the study, 56 (9.7%) delivered SGA10. SGA10 pregnancies had a significantly smaller PV, PQ, MPD, and mean placental diameter and higher PMI compared with normal pregnancies (P < .001 for each). Each placental measure remained significantly associated with SGA10 after adjusting for confounders and significantly improved the performance of the model using clinical variables alone (P < .04 for each) with adjusted areas under the curve ranging from 0.71 to 0.74. Uterine artery pulsatility index did not remain significantly associated with SGA10 after adjusting for confounders (P = .06). Placental growth factor was significantly lower in SGA10 pregnancies (P = .02) and remained significant in adjusted models but failed to significantly improve the predictive performance of the models as measured by area under the curve (P > .3). Placental protein-13 was not associated with SGA10 (P = .99). CONCLUSION: Direct assessment of placental size and shape with 3-dimensional ultrasound can serve as the foundation upon which to build a multivariable model for the early prediction of SGA.

Maternal serum serpin B7 is associated with early spontaneous preterm birth.

OBJECTIVE: We sought to identify serum biomarkers of early spontaneous preterm birth (SPTB) using semiquantitative proteomic analyses. STUDY DESIGN: This was a nested case-control study of pregnant women with previous SPTB. Maternal serum was collected at 19-24 and 28-32 weeks' gestation, and analyzed by liquid chromatography-multiple reaction monitoring/mass spectrometry. Targeted and shotgun proteomics identified 31 candidate proteins that were differentially expressed in pooled serum samples from spontaneous preterm (cases [<34 weeks]) and term (controls) deliveries. Candidate protein expression was compared in individual serum samples between cases and controls matched by age and race groups, and clinical site. Protein expression was verified by Western blot in the placenta and fetal membranes from cases and controls. RESULTS: Serum samples were available for 35 cases and 35 controls at 19-24 weeks, and 16 cases and 16 controls at 28-32 weeks. One protein, serpin B7, yielded serum concentrations that differed between cases and controls. The mean concentration of serpin B7 at 28-32 weeks was 1.5-fold higher in women with subsequent preterm deliveries compared to controls; there was no difference at 19-24 weeks. Higher levels of serpin B7 at both gestational age windows were associated with a shorter interval to delivery, and higher levels of serpin B7 in samples from 28-32 weeks were associated with a lower gestational age at delivery. Western blotting identified serpin B7 protein in placenta, amnion, and chorion from cases and controls. CONCLUSION: Targeted and shotgun serum proteomics analyses associated 1 protein, serpin B7, with early SPTB. Our results require validation in other cohorts and analysis of the possible mechanistic role of serpin B7 in parturition.

Extracellular vesicles alter trophoblast function in pregnancies complicated by COVID-19.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and resulting coronavirus disease (COVID-19) causes placental dysfunction, which increases the risk of adverse pregnancy outcomes. While abnormal placental pathology resulting from COVID-19 is common, direct infection of the placenta is rare. This suggests that pathophysiology associated with maternal COVID-19, rather than direct placental infection, is responsible for placental dysfunction and alteration of the placental transcriptome. We hypothesized that maternal circulating extracellular vesicles (EVs), altered by COVID-19 during pregnancy, contribute to placental dysfunction. To examine this hypothesis, we characterized maternal circulating EVs from pregnancies complicated by COVID-19 and tested their effects on trophoblast cell physiology in vitro . We found that the gestational timing of COVID-19 is a major determinant of circulating EV function and cargo. In vitro trophoblast exposure to EVs isolated from patients with an active infection at the time of delivery, but not EVs isolated from Controls, altered key trophoblast functions including hormone production and invasion. Thus, circulating EVs from participants with an active infection, both symptomatic and asymptomatic cases, can disrupt vital trophoblast functions. EV cargo differed between participants with COVID-19 and Controls, which may contribute to the disruption of the placental transcriptome and morphology. Our findings show that COVID-19 can have effects throughout pregnancy on circulating EVs and circulating EVs are likely to participate in placental dysfunction induced by COVID-19.

[Intraoperative nursing in bariatric surgery: integrative review]. / Enfermagem transoperatória nas cirurgias de redução de peso: revisão integrativa da literatura.

Integrative review of scientific literature study to identify and analyze the production of knowledge about clinical advances in security needs of patients during the intraoperative period of bariatric surgery. It was based on 12 selected studies in electronic databases, with descriptors previously defined. Except for two studies, the specific content of this production was composed of the general context of perioperative care. The studies highlight the possible state of the art of nursing activities on these needs, which are well established, including recommendations by several guidelines. However, they are fundamentally based on the science of traditional clinical practice through the development of rational judgments issued by experts. It concludes for the relevance of primary studies to evaluate the impact and resolution of the identified resources to answer those needs, as well as improving or generating other innovative features and identification of new needs.

[Skin lesions in the intraoperative period of cardiac surgery: incidence and characterization]. / Lesões de pele no intra-operatório de cirurgia cardíaca: incidência e caracterização.

The main objective of this exploratory, descriptive cohort study was to verify the incidence of patients submitted to cardiac surgery who developed skin lesions during the intraoperative period, and characterize the lesions. Data collection was performed at the Surgery Department of a public teaching hospital, of tertiary health care, mostly surgical, specialized in cardiology, and located in São Paulo. The study sample consisted of 182 patients. The study was performed with a significant p (<0.05) in nonparametric statistical tests. The incidence found for patients submitted to cardiac surgery who developed skin lesions due to the preoperative period was 20.9%. It was observed that 19.2% of lesions were Pressure Ulcers (PU) in stage I; 1.1% of lesions were abrasive; 1.1% incisive; 0.5% lacerative; 0.5% superficial electrical burns; and 0.5% PU in stage II.

Eating disorders and psychosis: a review and case report.

INTRODUCTION: The interplay between eating disorders and psychosis is a challenging field to which little attention has been paid. Its study raises conceptual and methodological questions in both areas, making the diagnosis and management of patients difficult. Such questions are addressed and illustrated with a review and case report. METHODS: The authors present the case of a woman with Anorexia Nervosa and with comorbid Shared Psychotic Disorder, based on a literature review regarding the comorbidity between eating disorders and psychosis. The authors conducted a non-systematic review by searching the PubMed database, using the Mesh Terms "anorexia nervosa", "bulimia nervosa", "comorbidity" and "psychotic disorders". RESULTS: The findings suggest that studies on the subject are limited by issues regarding data on the prevalence of comorbidities, phenomenological aspects of eating disorders, and the interface and integration with psychotic symptoms. CONCLUSIONS: The case presented illustrates the difficulties in managing a patient with a comorbid eating disorder and psychosis. In order to ensure a rigorous assessment of both psychotic and eating disorder symptoms, the focus should be on the pattern of appearance or emergence of symptoms, their phenomenology, clinical and family background of the patient, and clinical status on follow-up.

SARS-CoV-2 Seroprevalence Among Parturient Women.

Limited data are available for pregnant women affected by SARS-CoV-2. Serological tests are critically important to determine exposure and immunity to SARS-CoV-2 within both individuals and populations. We completed SARS-CoV-2 serological testing of 1,293 parturient women at two centers in Philadelphia from April 4 to June 3, 2020. We tested 834 pre-pandemic samples collected in 2019 and 15 samples from COVID-19 recovered donors to validate our assay, which has a ~1% false positive rate. We found 80/1,293 (6.2%) of parturient women possessed IgG and/or IgM SARS-CoV-2-specific antibodies. We found race/ethnicity differences in seroprevalence rates, with higher rates in Black/non-Hispanic and Hispanic/Latino women. Of the 72 seropositive women who also received nasopharyngeal polymerase chain reaction testing during pregnancy, 46 (64%) were positive. Continued serologic surveillance among pregnant women may inform perinatal clinical practices and can potentially be used to estimate seroprevalence within the community.

SARS-CoV-2 seroprevalence among parturient women in Philadelphia.

Limited data are available for pregnant women affected by SARS-CoV-2. Serological tests are critically important for determining SARS-CoV-2 exposures within both individuals and populations. We validated a SARS-CoV-2 spike receptor binding domain serological test using 834 pre-pandemic samples and 31 samples from COVID-19 recovered donors. We then completed SARS-CoV-2 serological testing of 1,293 parturient women at two centers in Philadelphia from April 4 to June 3, 2020. We found 80/1,293 (6.2%) of parturient women possessed IgG and/or IgM SARS-CoV-2-specific antibodies. We found race/ethnicity differences in seroprevalence rates, with higher rates in Black/non-Hispanic and Hispanic/Latino women. Of the 72 seropositive women who also received nasopharyngeal polymerase chain reaction testing during pregnancy, 46 (64%) were positive. Continued serologic surveillance among pregnant women may inform perinatal clinical practices and can potentially be used to estimate exposure to SARS-CoV-2 within the community.

Assessing beliefs and attitudes towards antipsychotic medication from a recovery-based perspective: Psychometric properties of a new scale.

Antipsychotic medication non-adherence is a complex and multifaceted problem that may hinder recovery in psychosis-spectrum disorders. Therefore, it warrants an early and comprehensive assessment. Current self-report measures focus entirely on behavioral and attitudinal barriers to adherence, failing to provide insight about key psychosocial drivers such as shame and stigma that may also account for non-adherence. This study's main goals were to develop a brief scale for measuring antipsychotic (non)-adherence and associated intra and interpersonal barriers (Antipsychotic Medication Beliefs and Attitudes Scale - AMBAS), and explore its psychometric properties. One hundred and seventy participants with a psychosis-spectrum disorder were recruited and filled in a battery of self-report measures. Exploratory factor analysis supported a two-factor solution, with one factor tapping the influence of different barriers to medication adherence and other factor encompassing perceived positive effects of medication. The scale presented good reliability and convergent validity as evidenced by significant moderate associations with the Medication Adherence Rating Scale. Although in need for further study, AMBAS seems a valid and reliable measure to assess antipsychotic (non)-adherence and underlying behavioral and psychosocial drivers. With replication, AMBAS might be a useful measure that could be used in different clinical and research settings.

Lack of detection of a human placenta microbiome in samples from preterm and term deliveries.

BACKGROUND: Historically, the human womb has been thought to be sterile in healthy pregnancies, but this idea has been challenged by recent studies using DNA sequence-based methods, which have suggested that the womb is colonized with bacteria. For example, analysis of DNA from placenta samples yielded small proportions of microbial sequences which were proposed to represent normal bacterial colonization. However, an analysis by our group showed no distinction between background negative controls and placenta samples. Also supporting the idea that the womb is sterile is the observation that germ-free mammals can be generated by sterile delivery of neonates into a sterile isolator, after which neonates remain germ-free, which would seem to provide strong data in support of sterility of the womb. RESULTS: To probe this further and to investigate possible placental colonization associated with spontaneous preterm birth, we carried out another study comparing microbiota in placenta samples from 20 term and 20 spontaneous preterm deliveries. Both 16S rRNA marker gene sequencing and shotgun metagenomic sequencing were used to characterize placenta and control samples. We first quantified absolute amounts of bacterial 16S rRNA gene sequences using 16S rRNA gene quantitative PCR (qPCR). As in our previous study, levels were found to be low in the placenta samples and indistinguishable from negative controls. Analysis by DNA sequencing did not yield a placenta microbiome distinct from negative controls, either using marker gene sequencing as in our previous work, or with shotgun metagenomic sequencing. Several types of artifacts, including erroneous read classifications and barcode misattribution, needed to be identified and removed from the data to clarify this point. CONCLUSIONS: Our findings do not support the existence of a consistent placental microbiome, in either placenta from term deliveries or spontaneous preterm births.

Enabling precision medicine in neonatology, an integrated repository for preterm birth research.

Preterm birth, or the delivery of an infant prior to 37 weeks of gestation, is a significant cause of infant morbidity and mortality. In the last decade, the advent and continued development of molecular profiling technologies has enabled researchers to generate vast amount of 'omics' data, which together with integrative computational approaches, can help refine the current knowledge about disease mechanisms, diagnostics, and therapeutics. Here we describe the March of Dimes' Database for Preterm Birth Research (http://www.immport.org/resources/mod), a unique resource that contains a variety of 'omics' datasets related to preterm birth. The database is open publicly, and as of January 2018, links 13 molecular studies with data across tens of thousands of patients from 6 measurement modalities. The data in the repository are highly diverse and include genomic, transcriptomic, immunological, and microbiome data. Relevant datasets are augmented with additional molecular characterizations of almost 25,000 biological samples from public databases. We believe our data-sharing efforts will lead to enhanced research collaborations and coordination accelerating the overall pace of discovery in preterm birth research.

Author Correction: Enabling precision medicine in neonatology, an integrated repository for preterm birth research.

The original version of the Data Descriptor contained errors in the author list and affiliations. Rita Leite's first name was misspelled as "Rite" and affiliations 4 and 5 were incorrectly swapped. In addition, members of the March of Dimes Prematurity Research Center consortium were not listed in the agreed positions within the author list. These errors have now been corrected in the HTML and PDF versions.

A hemoglobin fragment found in cervicovaginal fluid from women in labor potentiates the action of agents that promote contraction of smooth muscle cells.

We employed a proteomic approach to search for peptides that have a physiological role in labor. Cervicovaginal secretions were collected at term from women in labor and women at term not in labor. Samples were spotted onto weak cation exchange chips (WCX-2) and analyzed using Surface-Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry (SELDI-TOF-MS). Spectra were obtained for each sample and Biomarker Wizard analysis revealed 25 peaks that had significantly different peak intensity between the labor and non-laboring women. The sequences of five peaks that were significantly elevated in the labor cohort were determined using Protein Chip Interface Quadruple Time-of-Flight Mass Spectrometry (PCI-QTOF-MS). All of these peaks were identified as fragments of alpha or beta-hemoglobin (Hb). A 2.022 kDa fragment of alpha-Hb (amino acids 110-128, NH2-AAHLPAEFTPAVHASLDKF-COOH) was found to potentiate smooth muscle cell contraction in response to bradykinin, oxytocin and prostaglandin-F2alpha. This peptide may promote vasoconstriction and augment normal labor through enhancing the action of uterotonins.

Comparison of placenta samples with contamination controls does not provide evidence for a distinct placenta microbiota.

BACKGROUND: Recent studies have suggested that bacteria associated with the placenta-a "placental microbiome"-may be important in reproductive health and disease. However, a challenge in working with specimens with low bacterial biomass, such as placental samples, is that some or all of the bacterial DNA may derive from contamination in dust or commercial reagents. To investigate this, we compared placental samples from healthy deliveries to a matched set of contamination controls, as well as to oral and vaginal samples from the same women. RESULTS: We quantified total 16S rRNA gene copies using quantitative PCR and found that placental samples and negative controls contained low and indistinguishable copy numbers. Oral and vaginal swab samples, in contrast, showed higher copy numbers. We carried out 16S rRNA gene sequencing and community analysis and found no separation between communities from placental samples and contamination controls, though oral and vaginal samples showed characteristic, distinctive composition. Two different DNA purification methods were compared with similar conclusions, though the composition of the contamination background differed. Authentically present microbiota should yield mostly similar results regardless of the purification method used-this was seen for oral samples, but no placental bacterial lineages were (1) shared between extraction methods, (2) present at >1 % of the total, and (3) present at greater abundance in placental samples than contamination controls. CONCLUSIONS: We conclude that for this sample set, using the methods described, we could not distinguish between placental samples and contamination introduced during DNA purification.

Tumor necrosis factor-alpha suppresses the expression of steroid receptor coactivator-1 and -2: a possible mechanism contributing to changes in steroid hormone responsiveness.

Inflammation is associated in some tissues with diminished responsiveness to steroid hormone action. We hypothesized that proinflammatory cytokines alter steroid hormone sensitivity, in part, by reducing levels of key nuclear receptor coactivators. Treatment of cultured human uterine smooth muscle cells (UtSMC) with TNF-alpha significantly reduced mRNA for the coactivators, SRC-1 (42%, P<0.01) and 2 (47%, P<0.03), and diminished the respective protein levels, but did not significantly alter the mRNAs encoding SRC-3, CBP and the corepressors, NCoR and SMRT; or progesterone receptor protein levels. To assess TNF-alpha effects on steroid hormone-mediated transcriptional activity, UtSMC were transfected with progesterone receptor B (PR-B) and a model PRE2-luciferase reporter construct. Transfected UtSMC were treated with progesterone alone or in the presence of TNF-alpha, and assayed for luciferase activity. TNF-alpha (10 ng/ml) diminished progesterone-stimulated PR-B-mediated transactivation by approximately 60% (P<0.02). The TNF-alpha-dependent decrease in PRE-luciferase activity was fully prevented by cotransfection with SRC-2, and partially prevented with exogenous SRC-1. In conclusion, TNF-alpha impairs progesterone-stimulated PR-B-mediated transactivation, and these effects appear to be due, in part, to reduced expression of SRC-1 and -2, which is a novel mechanism by which inflammation can functionally block steroid hormone action.