HIV-1 Seroprevalence among Pregnant Women in Rural Uganda: A Longitudinal Study over Fifteen Years

Introduction: In order to determine the development of the prevalence of HIV infection in rural Western Uganda, data of epidemiological studies conducted in 2001 and 2007 were compared to study data from 1993. Methods: In 2001 (n = 466) and in 2007 (n = 486), one group each of clinically healthy pregnant women of a local prenatal care department were enrolled in the study and anonymously screened for HIV-1. For both groups, informed consent was obtained prior to enrolment. Testing for HIV was done by enzyme-linked immunosorbent assay (ELISA) and confirmed by Western blot. In addition, age and antibodies against syphilis were determined as risk factors of HIV infection. Results: The seroprevalence of HIV-1 infection did not decrease significantly over this time period, dropping from 28.3 to 25.1% between 2001 and 2007, but the prevalence of syphilis antibodies decreased from 27.9 to 11.1%. The data of 2001 and 2007 were compared to a third cohort from 1993, in which 21.5% of pregnant women were HIV-1-positive and 31.1% were Treponema pallidum hemagglutination assay (TPHA)-positive. Conclusion: The current prevalence of HIV-1 infection in Uganda is still high and there is a need for further promotion of HIV prevention and control services

Cytokine profiles of umbilical cord blood mononuclear cells upon in vitro stimulation with lipopolysaccharides of different vaginal gram-negative bacteria

Inflammatory immune responses induced by lipopolysaccharides (LPS) of gram-negative bacteria play an important role in the pathogenesis of preterm labor and delivery, and in neonatal disorders. To better characterize LPS-induced inflammatory response, we determined the cytokine profile of umbilical cord blood mononuclear cells (UBMC) stimulated with LPS of seven vaginal gram-negative bacteria commonly found in pregnant women with preterm labor and preterm rupture of membrane. UBMC from ten newborns of healthy volunteer mothers were stimulated with purified LPS of Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Proteus mirabilis, Acinetobacter calcoaceticus, Citrobacter freundii, and Pseudomonas aeruginosa. UBMC supernatants were tested for the presence of secreted pro-inflammatory cytokines (IL-6, IL-1β, TNF), anti-inflammatory cytokine (IL-10), TH1-type cytokines (IL-12, IFN-γ), and chemokines (IL-8, MIP-1α, MIP-1β, MCP-1) by Luminex technology. The ten cytokines were differentially induced by the LPS variants. LPS of E. coli and E. aerogenes showed the strongest stimulatory activity and P. aeruginosa the lowest. Interestingly, the ability of UBMC to respond to LPS varied greatly among donors, suggesting a strong individual heterogeneity in LPS-triggered inflammatory response

Comparative purification and characterization of hepatitis B virus-like particles produced by recombinant vaccinia viruses in human hepatoma cells and human primary hepatocytes

This study describes the comparative expression and purification of hepatitis B surface antigen (HBsAg) particles produced upon infection of human primary hepatocytes and human hepatoma cell lines (HuH-7 and HepG2) with recombinant vaccinia viruses. The highest levels of HBsAg expression were found in HuH-7 hepatoma cells following infection with recombinant vaccinia viruses, which contain the S gene under control of a 7.5 k-promoter. Four different methods for purification of the HBsAg particles were examined: isopycnic ultracentrifugation, sucrose cushion sedimentation, isocratic column gel filtration, and binding to anti-HBs-coated microparticles. The highest degree of purity of HBsAg particles was reached by the method based on anti-HBs-coated microparticles. The resulting product was > 98% pure. Biochemical analysis and characterization of purified HBsAg particles were performed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, and electron microscopy. The HBsAg, purified from human hepatoma cell lines and from human primary hepatocytes, consisted of both the non-glycosylated (p25) and the glycosylated (gp27) form and assembled into typical 22-nm particles, and thus may be of great interest and importance for research, diagnostics, and medical treatments

Revisiting promyelocytic leukemia protein targeting by human cytomegalovirus immediate-early protein 1

This work was supported by a grant (MR/P022146/1) from the Medical Research Council (https://mrc.ukri.org) to MMN, a grant (T16/28) from Tenovus Scotland (https://tenovus-scotland.org.uk) to CP, a European Union Erasmus+ grant (https://www.erasmusplus.org.uk) to BW and the Wellcome Trust Institutional Strategic Support Fund (https://wellcome.ac.uk) to CP and MMN.Promyelocytic leukemia (PML) bodies are nuclear organelles implicated in intrinsic and innate antiviral defense. The eponymous PML proteins, central to the self-organization of PML bodies, and other restriction factors found in these organelles are common targets of viral antagonism. The 72-kDa immediate-early protein 1 (IE1) is the principal antagonist of PML bodies encoded by the human cytomegalovirus (hCMV). IE1 is believed to disrupt PML bodies by inhibiting PML SUMOylation, while PML was proposed to act as an E3 ligase for IE1 SUMOylation. PML targeting by IE1 is considered to be crucial for hCMV replication at low multiplicities of infection, in part via counteracting antiviral gene induction linked to the cellular interferon (IFN) response. However, current concepts of IE1-PML interaction are largely derived from mutant IE1 proteins known or predicted to be metabolically unstable and globally misfolded. We performed systematic clustered charge-to-alanine scanning mutagenesis and identified a stable IE1 mutant protein (IE1cc172-176) with wild-type characteristics except for neither interacting with PML proteins nor inhibiting PML SUMOylation. Consequently, IE1cc172-176 does not associate with PML bodies and is selectively impaired for disrupting these organelles. Surprisingly, functional analysis of IE1cc172-176 revealed that the protein is hypermodified by mixed SUMO chains and that IE1 SUMOylation depends on nucleosome rather than PML binding. Furthermore, a mutant hCMV expressing IE1cc172-176 was only slightly attenuated compared to an IE1-null virus even at low multiplicities of infection. Finally, hCMV-induced expression of cytokine and IFN-stimulated genes turned out to be reduced rather than increased in the presence of IE1cc172-176 relative to wild-type IE1. Our findings challenge present views on the relationship of IE1 with PML and the role of PML in hCMV replication. This study also provides initial evidence for the idea that disruption of PML bodies upon viral infection is linked to activation rather than inhibition of innate immunity.Publisher PDFPeer reviewe

Usage of the wearable cardioverter-defibrillator during pregnancy

Background Pregnancy can trigger or aggravate the risk for life-threating arrhythmias in cardiac diseases. Pregnancy is associated with reluctance for implantable cardioverter-defibrillators (ICD) due to concerns about radiation. Thus, the wearable cardioverter-defibrillator (WCD) might be an option during pregnancy. Aim of the study was to collect experiences about the use of WCD in pregnancy. Methods and results This study retrospectively included eight women who received a WCD during pregnancy. They suffered from ventricular tachycardia (VT) without known cardiac disease (n = 3), Brugada syndrome (n = 1), hypertrophic cardiomyopathy (n = 1), dilated cardiomyopathy (n = 1), non-compaction (n = 1), and survived sudden cardiac arrest during a preceding pregnancy (n = 1). WCD usage was started between 13 and 28 weeks of gestation. WCD wearing period ranged from 3 days to 30.9 weeks, WCD wearing time ranged from 13.0 to 23.7 h per day. Two women (25%) abandoned WCD already during pregnancy. Neither appropriate nor inappropriate WCD shocks were recorded. Antiarrhythmic management included beta-blockers (n = 5) and flecainide (n = 2). After delivery, ICD were implanted (n = 4), refused (n = 2) and estimated not necessary after successful catheter ablation (n = 2). Conclusion Uneventful pregnancy is possible in women at risk for sudden cardiac death by interdisciplinary monitoring and diligent pharmacotherapy protected by the WCD. Since no WCD shocks were recorded, the effectiveness of WCD during pregnancy is still unclear. However, arrhythmia detection by WCD was very good despite the changed anatomy in pregnancy. Nevertheless, further studies are necessary to assess effectiveness of WCD in pregnant women. Furthermore, efforts should be made to increase the wearing adherence of WCD during pregnancy

Aberrant DNA methylation patterns in microsatellite stable human colorectal cancers define a new marker panel for the CpG island methylator phenotype

A distinct group of colorectal carcinomas (CRCs) referred to as the “CpG island methylator phenotype” (CIMP) shows an extremely high incidence of de novo DNA methylation and may share common pathological, clinical or molecular features. However, there is limited consensus about which CpG islands (CGIs) define a CIMP, particularly in microsatellite stable (MSS) carcinomas. To study this phenotype in a systematic manner, we analyzed genome-wide CGI DNA methylation profiles of 19 MSS CRC using methyl-CpG immunoprecipitation (MCIp) and hybridization on 244K CGI oligonucleotide microarrays, determined KRAS and BRAF mutation status and compared disease-related DNA methylation changes to chromosomal instability as detected by microarray-based comparative genomic hybridization. Results were validated using mass spectrometry analysis of bisulfite-converted DNA at a subset of 76 individual CGIs in 120 CRC and 43 matched normal tissue samples. Both genome-wide profiling and CpG methylation fine mapping segregated a group of CRC showing pronounced and frequent de novo DNA methylation of a distinct group of CGIs that only partially overlapped with previously established classifiers. The CIMP group defined in our study revealed significant association with colon localization, either KRAS or BRAF mutation, and mostly minor chromosomal losses but no association with known histopathological features. Our data provide a basis for defining novel marker panels that may enable a more reliable classification of CIMP in all CRCs, independently of the MS status

Changes in Maternal Body Mass Index, Weight Gain and Outcome of Singleton Pregnancies from 2000 to 2015

Introduction Maternal obesity and excessive gestational weight gain (GWG) affect the outcomes of women and their offspring. Our aim was to evaluate population-based data from Germany. Material and Methods Data from 583633/791514 mother-child pairs obtained from the perinatal database in Hesse for the period from 2000 to 2015 were used after excluding incomplete or non-plausible datasets. Early-stage pregnancy maternal body mass index (BMI) and GWG were evaluated. Significant outcome changes were calculated using linear or logistic regression models. Results The mean maternal age increased from 29.9 to 31.28 years; GWG increased from 445.1 to 457.2 g/week (p < 0.01). Similarly, rates for both overweight and obesity rose from 31.5 to 37.5% (p < 0.001). Cesarean section rates rose from 22.8 to 33.2% (p < 0.001) and rates of postpartum hemorrhage increased from 0.6 to 1% (p < 0.001). There was no significant change in the rates for stillbirth or perinatal mortality (p = 0.92 and p = 0.53 respectively), but there was an increase in the rates of admissions to neonatal intensive care units from 7.8 to 9.5% (p < 0.0001). The percentage of newborns with an Apgar score of < 7 at 5 minutes increased from 1 to 1.1% (p < 0.01) and the rate of neonates with an umbilical artery pH of < 7.1 rose from 1.7 to 2.4% (p < 0.01). Conclusions In just 15 years, pre-pregnancy BMI and GWG rates of women with singleton pregnancies have increased, and this increase has been accompanied by a significant rise in the rate of cesarean sections and a significant worsening of short-term maternal and neonatal outcomes. It is time to discuss the risks and the short-term and more worrying long-term consequences for mothers and their offspring and the future impact on our healthcare system

Perinatal Gram-Positive Bacteria Exposure Elicits Distinct Cytokine Responses In Vitro

During pregnancy, infections caused by the gram-positive bacteria Enterococcus faecalis (E. faecalis), Streptococcus agalacticae (S. agalacticae), and Staphylococcus aureus (S. aureus) are major reasons for preterm labor, neonatal prematurity, meningitis, or sepsis. Here, we propose cytokine responses to bacterial infections by the immature perinatal immune system as central players in the pathogenesis of preterm birth and neonatal sepsis. We aimed to close the gap in knowledge about such cytokine responses by stimulating freshly isolated umbilical blood mononuclear cells (UBMC) with lysates of E. faecalis, S. agalacticae, and S. aureus collected from pregnant women in preterm labor. Bacterial lysates and, principally, S. aureus and S. agalacticae distinctly triggered most of the eleven inflammatory, anti-inflammatory, TH1/TH2 cytokines, and chemokines quantified in UBMC culture media. Chemokines depicted the most robust induction. Among them, MIP-1 beta was further enhanced in UBMC from female compered to male newborn infants. Due to its stability and high levels, we investigated the diagnostic value of IL-8. IL-8 was critically upregulated in cord blood of preterm neonates suffering from infections compared to gestational age-matched controls. Our results provide novel clues about perinatal immunity, underscoring a potential value of IL-8 for the timely detection of infections and suggesting that MIP-1 beta constitutes an early determinant of sex-specific immunity, which may contribute, e.g., to male's vulnerability to preterm birth

Perinatal Gram-Positive Bacteria Exposure Elicits Distinct Cytokine Responses In Vitro

During pregnancy, infections caused by the gram-positive bacteria Enterococcus faecalis (E. faecalis), Streptococcus agalacticae (S. agalacticae), and Staphylococcus aureus (S. aureus) are major reasons for preterm labor, neonatal prematurity, meningitis, or sepsis. Here, we propose cytokine responses to bacterial infections by the immature perinatal immune system as central players in the pathogenesis of preterm birth and neonatal sepsis. We aimed to close the gap in knowledge about such cytokine responses by stimulating freshly isolated umbilical blood mononuclear cells (UBMC) with lysates of E. faecalis, S. agalacticae, and S. aureus collected from pregnant women in preterm labor. Bacterial lysates and, principally, S. aureus and S. agalacticae distinctly triggered most of the eleven inflammatory, anti-inflammatory, TH1/TH2 cytokines, and chemokines quantified in UBMC culture media. Chemokines depicted the most robust induction. Among them, MIP-1&beta; was further enhanced in UBMC from female compered to male newborn infants. Due to its stability and high levels, we investigated the diagnostic value of IL-8. IL-8 was critically upregulated in cord blood of preterm neonates suffering from infections compared to gestational age-matched controls. Our results provide novel clues about perinatal immunity, underscoring a potential value of IL-8 for the timely detection of infections and suggesting that MIP-1&beta; constitutes an early determinant of sex-specific immunity, which may contribute, e.g., to male&rsquo;s vulnerability to preterm birth