Gene Transfer to Chicks Using Lentiviral Vectors Administered via the Embryonic Chorioallantoic Membrane

The lack of affordable techniques for gene transfer in birds has inhibited the advancement of molecular studies in avian species. Here we demonstrate a new approach for introducing genes into chicken somatic tissues by administration of a lentiviral vector, derived from the feline immunodeficiency virus (FIV), into the chorioallantoic membrane (CAM) of chick embryos on embryonic day 11. The FIV-derived vectors carried yellow fluorescent protein (YFP) or recombinant alpha-melanocyte-stimulating hormone (α-MSH) genes, driven by the cytomegalovirus (CMV) promoter. Transgene expression, detected in chicks 2 days after hatch by quantitative real-time PCR, was mostly observed in the liver and spleen. Lower expression levels were also detected in the brain, kidney, heart and breast muscle. Immunofluorescence and flow cytometry analyses confirmed transgene expression in chick tissues at the protein level, demonstrating a transduction efficiency of ∼0.46% of liver cells. Integration of the viral vector into the chicken genome was demonstrated using genomic repetitive (CR1)-PCR amplification. Viability and stability of the transduced cells was confirmed using terminal deoxynucleotidyl transferase (dUTP) nick end labeling (TUNEL) assay, immunostaining with anti-proliferating cell nuclear antigen (anti-PCNA), and detection of transgene expression 51 days post transduction. Our approach led to only 9% drop in hatching efficiency compared to non-injected embryos, and all of the hatched chicks expressed the transgenes. We suggest that the transduction efficiency of FIV vectors combined with the accessibility of the CAM vasculature as a delivery route comprise a new powerful and practical approach for gene delivery into somatic tissues of chickens. Most relevant is the efficient transduction of the liver, which specializes in the production and secretion of proteins, thereby providing an optimal target for prolonged study of secreted hormones and peptides

Short Interpregnancy Interval Following a Multifetal Pregnancy: Maternal and Neonatal Outcomes

Objective: To evaluate the maternal and neonatal outcomes of women with short interpregnancy intervals (IPI p = 0.03). Conclusion: Short IPI following a multifetal gestation is associated with an increased risk for preterm birth in subsequent singleton pregnancy

Recurrent unintended pregnancies among young unmarried women serving in the Israeli military

Abstract Background Unintended pregnancy is a major public health problem with known risk factors, however, little is known about the prevalence of variables associated with recurrent unintended pregnancy (RUP) among young, unmarried women. Methods A retrospective cohort study of unmarried women aged 18–21 serving in the Israeli military between 2013 and 2015. Multivariable logistic regression analysis was used to examine associations between RUP and women’s education, IQ, immigration status, country of origin, socioeconomic status and history of psychiatric illness. Results Of 129,638 women drafted by the Israeli military during the study period, 1720 women with unintended pregnancies had a follow up period of at least a year. Three hundred and eighty-nine of them had RUP (22.6%). Multivariable models comparing women with no unintended pregnancies and women with RUP revealed that RUP was more common among (adjusted relative risk; 95% confidence interval) women who had not graduated from high school (6.9; 4.99–9.55), who had low (90–99) IQ scores (3.9; 2.88–5.39) those reporting Africa as the country of origin (2.5; 1.37–4.59) and those from a lower socioeconomic neighborhood (1.6; 1.18–2.05). Multivariate regression modeling comparing women with single unintended pregnancies and women with RUPs showed that recurrent unintended pregnancy was more common among women who had not graduated from high school (3.2; 2.04–4.84) and those who had a low (90–99) IQ score (1.9; 1.32–2.61). Conclusion Rate of RUP is high among women serving in the Israeli military. These women have unique epidemiological characteristics. This may serve in identifying populations at high risk and thus may enable policy maker to offer at least to this population Long-Acting Reversible Contraception (LARC) methods. We encourage policy makers to consider the provision of LARC methods to all servicewomen who had an unintended pregnancy

Prolonged operative time of repeat cesarean is a risk marker for post-operative maternal complications

Abstract Background Repeat cesarean delivery (CD) accounts for approximately 15% of all annual deliveries in the US with an estimated 656,250 operations per year. We aimed to study whether prolonged operative time (OT; skin incision to closure) is a risk marker for post-operative maternal complications among women undergoing repeat CD. Methods We conducted a cross-sectional retrospective study in a single tertiary center including all women who underwent repeat CD but excluding those with cesarean hysterectomy. Prolonged OT was defined as duration of CD longer than the 90th percentile duration on record for each specific surgeon in order to correct for technique differences between surgeons. Bi-variate analysis was used to study the association of prolonged OT with each one of the following maternal complications: post-operative blood transfusion, prolonged maternal hospitalization (defined as hospitalization duration longer than 1 week post-CD), infection necessitating antibiotics, re-laparotomy within 7 days post-CD, and re-admission within 42 days post-CD. A multivariate regression analysis was performed controlling for maternal age, ethnicity, parity, number of fetus, gestational age at delivery, trial of labor after cesarean, anesthesia, and number of previous CDs. The adjusted odd ratio was calculated for each complication independently and for a composite adverse maternal outcome defined as any one of the above. Results A total of 6507 repeat CDs were included; prolonged OT was highly associated (P value < 0.000) with: post-operative blood transfusion (4.4% vs. 1.5%), prolonged hospitalization (8.4% vs. 4.0%), infection necessitating antibiotics (2% vs. 1%), and readmission (1.8% vs. 0.8%) when compared to control. The composite adverse maternal outcome was also associated with prolonged OT (20.2% vs. 11.2%, p < 0.000). These correlations remained statistically significant in the multivariate regression analysis when controlling for confounders. Conclusions Among women undergoing repeat CD, prolonged OT (reflecting CD duration greater than 90th percentile for the specific surgeon) is a risk marker for post-operative maternal complications

Maternal and Neonatal Outcomes of Women Conceived Less Than 6 Months after First Trimester Dilation and Curettage

Objective: To evaluate the maternal and neonatal outcomes of pregnancies conceived ≤6 months after first trimester (p = 0.11] for preterm birth. Conclusion: Watchful waiting or the medical treatment of a first trimester missed abortion present no more risks than D&C to pregnancies conceived within six months of the index miscarriage. Further studies in other settings to strengthen these findings are needed

A Retrospective Study of the Proportion of Women at High and Low Risk of Intrauterine Infection Meeting Sepsis Criteria

The Surviving Sepsis Campaign recently recommended that qSOFA not be used as a single parameter for identification of sepsis. Thus, we evaluated the efficacy of SIRS and qSOFA scores in identifying intrauterine infection. This case&ndash;control study evaluates SIRS and qSOFA criteria fulfillment in preterm premature rupture of membranes (n = 453)&mdash;at high infection risk&mdash;versus elective cesarean&mdash;at low infection risk (n = 2004); secondary outcomes included intrauterine infection and positive culture rates. At admission, 14.8% of the study group and 4.6% of control met SIRS criteria (p = 0.001), as did 12.5% and 5.5% on post-operation day (POD) 1 (p = 0.001), with no significant differences on POD 0 or 2. Medical records did not suffice for qSOFA calculation. In the study group, more cultures (29.8% versus 1.9%&mdash;cervix; 27.4% versus 1.1%&mdash;placenta; 7.5% versus 1.7%&mdash;blood; p = 0.001&mdash;all differences) and positive cultures (5.5% versus 3.0%&mdash;urine&mdash;p = 0.008; 4.2% versus 0.2%&mdash;cervix&mdash;p = 0.001; 7.3% versus 0.0%&mdash;placenta&mdash;p = 0.001; 0.9% versus 0.1%&mdash;blood&mdash;p = 0.008) were obtained. Overall, 10.6% of the study group and 0.4% of control met the intrauterine infection criteria (p = 0.001). Though a significant difference was noted in SIRS criteria fulfillment in the study group versus control, there was considerable between-group overlap, questioning the utility of SIRS in intrauterine infection diagnosis. Furthermore, the qSOFA scores could not be assessed