Optimization of Propidium Monoazide qPCR (Viability-qPCR) to Quantify the Killing by the Gardnerella-Specific Endolysin PM-477, Directly in Vaginal Samples from Women with Bacterial Vaginosis.

Quantification of the number of living cells in biofilm or after eradication treatments of biofilm, is problematic for different reasons. We assessed the performance of pre-treatment of DNA, planktonic cells and ex vivo vaginal biofilms of Gardnerella with propidium monoazide (PMAxx) to prevent qPCR-based amplification of DNA from killed cells (viability-qPCR). Standard PMAxx treatment did not completely inactivate free DNA and did not affect living cells. While culture indicated that killing of planktonic cells by heat or by endolysin was complete, viability-qPCR assessed only log reductions of 1.73 and 0.32, respectively. Therefore, we improved the standard protocol by comparing different (combinations of) parameters, such as concentration of PMAxx, and repetition, duration and incubation conditions of treatment. The optimized PMAxx treatment condition for further experiments consisted of three cycles, each of: 15 min incubation on ice with 50 µM PMAxx, followed by 15 min-long light exposure. This protocol was validated for use in vaginal samples from women with bacterial vaginosis. Up to log2.2 reduction of Gardnerella cells after treatment with PM-477 was documented, despite the complex composition of the samples, which might have hampered the activity of PM-477 as well as the quantification of low loads by viability-qPCR.

Acute SARS-CoV-2 alpha variant infection leading to placental insufficiency and fetal distress.

The effect of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Alpha variant (also known as B.1.1.7 lineage, 20I/501Y.V1, the UK variant or VOC 202012/01) infection on pregnancy is currently unknown. We present a case of a 37-year-old woman admitted to our tertiary hospital at a gestational age of 29 weeks and 1 day because of oligohydramnios with reduced fetal movements for 10 days. About 20 days before admission, she tested positive for SARS-CoV-2 Alpha variant. The following day, due to abnormal cardiotocography, increased brain sparing, and absent end-diastolic flow in the umbilical artery, an urgent cesarean section was performed. The neonate had an uneventful admission to the neonatal intensive care unit. All neonatal samples proved negative for SARS-CoV-2. Pathological examination of the placenta revealed intervillous fibrin deposition, ischemic necrosis of villi and histiocytic intervillositis, corresponding with the SARS-CoV-2 placentitis triad. The placental tissue demonstrated a high viral load, possibly explaining the acute onset of placental insufficiently and subsequent fetal distress. This case demonstrates the importance of seeking medical care when experiencing reduced fetal movement in SARS-CoV-2 infected patients since acute infection can induce significant placental and subsequent fetal pathology.

Gestational weight gain and postpartum weight retention after bariatric surgery: data from a prospective cohort study.

BACKGROUND: It is unknown whether international guidelines on gestational weight gain can be used in pregnancies after bariatric surgery. OBJECTIVES: To investigate gestational weight gain, intrauterine growth, and postpartum weight retention in postbariatric women. SETTING: 8 Belgian hospitals. METHODS: Prospective data from 127 postbariatric pregnancies from September 2014 through October 2018. Patients were grouped according to achievement of 2009 Institute of Medicine (IOM) guidelines. RESULTS: In 127 patients with a mean age of 30.2 years (standard deviation [SD], 4.7), the mean gestational weight gain was 12.5 kg (SD, 6.7). Of these patients, 24% (30 of 127) showed insufficient weight gain, 20% (26 of 127) showed adequate weight gain, and 56% (71 of 127) showed excessive weight gain. Of 127 patients, 27 (21%) had small-for-gestational-age infants. This peaked in the group with insufficient weight gain (47%; 95% confidence interval [CI], 29%-65%; P < .001). The prevalence of large-for-gestational-age infants was comparable between groups, although highest in the group with excessive weight gain (0% in those with insufficient weight gain, 4% in those with adequate weight gain, and 8% in those with excessive weight gain). Preterm births were recorded more in patients with insufficient weight gain (23%; 95% CI, 8%-38%; P = .048). The mean amounts of postpartum weight retained were 4.0 kg (SD, 7.4) at 6 weeks and 3.0 kg (SD, 9.1) at 6 months. Weight retention at 6 weeks (7.1 kg; 95% CI, 5.5-8.7; P < .001) and 6 months (8.3 kg; 95% CI, 4.5-12.2; P < .001) was highest in women gaining excessive weight. CONCLUSION: Achievement of IOM guidelines is low in postbariatric pregnancies. Insufficient weight gain increases the risk for small-for-gestational-age babies. Excessive weight gain increases weight retention after delivery and could precipitate weight regain. After bariatric surgery, women should be encouraged to achieve IOM recommendations.