SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 186A

186A

Reproductive Sciences Vol. 25, Supplement 1, March 2018

becomes detectable at low levels at 18.5 dpc and is greatly elevated
immediately after term birth. We describe two methods, immunoblot
and immunostaining of the lung, to measure surfactant levels. We also
describe more thorough histology of the lung and skin that can also used
to provide deeper characterization.
CONCLUSION: We propose the adoption of standard reporting
guidelines and the use of fetal developmental markers to distinguish
between short gestation and preterm birth.

F-011
Nodal Deletion Increases Susceptibility to LPS-Induced Preterm Birth
in Mice. Taghreed Heba†. McGill University, Montreal, QC, Canada.
INTRODUCTION: Preterm birth remains the major cause of perinatal
mortality and morbidty worldwide, affecting up to 12% of pregnacies and
accounting for approximatly 75% of neonatal deaths but the mechanisms
and causes that underlie it are still largely unknown. Genteic predispostion
is known contributing factors to preterm birth. Maternal inflammation is
also an important risk factor that associates with spontaneous preterm
birth. Our lab has been studying the role of the Nodal signaling pathway
during pregnancy. Nodal is a morphogen that belongs to the (TGF-B)
superfamily. Nodal has been shown to play critical roles during embryonic
development and our lab has shown that it is also required during gestation,
implantation and the timing of parturition.
METHODS: We used a conditional uterine Nodal heterozygous knockout
mouse model to evaluate their susceptibility to inflammation during late
pregnancy after injection by a vey low dose of an inflammatory mediator
LPS. Furthermore, by using an RT2 Profiler PCR array, we examined
the expression of 84 genes involved in innate and adaptive immunity. In
addition, ELISA profiling system was used to verify the protein expression
level of several proinflammatory cytokines including (IL-1β), (IL-6),
(IL-12p70), (TNF-α), (IFN-γ) and an anti-inflammatory cytokines (IL10). An immunoflurescnce staining was used to show the localiziation
of macrophges and utrine natural killer cells (uNK) in placental tissue in
the late pregnancy. Our hypothesis that Nodal regulate the immune cells
in mouse placenta
RESULTS: We showed that Nodal heterozygous mice have a fertility
rate of only 50%. Injection of a low dose of LPS (1.4 mg/kg) caused 50%
of pregnant Nodal heterozygous knockout mice but 0% of control mice
to have preterm birth. Moreover, the RT2 Profiler PCR array showed a
significant upregulation of critical cytokines in the patrturtion cascade
in maternal and placental tissue at day 16.5 of pregnancy. Surprisingly,
ELISA showed most of the pro-inflammatory cytokines were elevated
in the Nodal heterozygous knockout mice compared to the control. An
immunoflurescnce staining, showed a significant increase in the number
of uterine NK cells and macrophages in the maternal tissue in Nodal
heterozygous knockout mice compared to controls.
CONCLUSION: NODAL may regulate immune cells in placental and
maternal tissue.

F-012
Women's Experiences of Incorporating Cervical Electrical Impedance
Spectroscopy into Conventional Clinical Assessment of Risk of
Preterm Birth. Victoria Stern†,2 Sarah Senbeto,2 Georgina Jones,1 Dilly O
Anumba*.2 1Leeds Beckett University, Leeds, United Kingdom; 2University
of Sheffield, Sheffield, United Kingdom.
INTRODUCTION: We have recently quantified differences in cervical
resistivity in pregnant women destined to deliver preterm (ECCLIPPx™
studies). A concurrent qualitative study was conducted to determine the
acceptability of Electrical Impedance Spectroscopy (EIS) to women.
During the main study, low (LR) and high risk (HR) women attended
visits at 20-22 weeks; HR women attended again at 26-28 weeks. All
participants underwent speculum examination to obtain swabs for
infection screening and fetal fibronectin (FFN) quantification, then EIS
measurements) followed by a transvaginal ultrasound scan to measure
cervical length (CL).
METHODS: The 21 participants (11 LR, 10 HR) in the acceptability
study were recruited over 12 months. Pre and post-visit questionnaires
utilising validated scales were used to assess anxiety (short form

Scientific Abstracts

Spielberger Stait-Trait Anxiety Inventory) and pain (short-form McGill
Pain Questionnaire); a visual analogue scale (VAS) was used to evaluate
the overall acceptability of the EIS procedure and appearance of the
spectroscopy device. The EQ-5D-5L instrument was used to evaluate
overall health outcomes after testing. Participants also attended for a
semi-structured qualitative interview with a research midwife to discuss
their experiences in detail.
RESULTS: The majority of women (66%) experienced a reduction in
anxiety after testing, with 23% reporting no change. One noted an increase
in anxiety. High risk women showed a non-significant trend to higher pretest anxiety (mean 34 vs 32 p=0.79), but greater post-test reduction (mean
reduction -7 vs -5 p= 0.63). Pain scores were low (mean visual analogue
score 0.85/10 range 0-3, mean present pain intensity 0.55/5 range 0-2)
with 'tender' and 'aching' being the most frequently selected sensory
descriptors. All participants felt EIS was acceptable for use in antenatal
care (mean acceptability score 0.55 on VAS, range 0-5 where 0=highly
acceptable) and the majority did not find the appearance of the EIS device
threatening (mean score 1.65 on VAS assessment of appearance, range 0-9
where 0=not threatening). Mean EQ-5D-5L score was 85 (range 55-100)
- no unexpected effects of testing on daily function were identified. The
qualitative interviews identified both test and clinician-specific elements
which affected women's experiences. Clear explanation of results and
measurement scales increased women's confidence in test results, and
the nature of doctor-patient interaction and the screening environment
significantly impacted women's experiences.
CONCLUSION: EIS is an acceptable preterm screening test for both low
and high risk women. Useful detailed impressions of the device appearance
obtained during qualitative interviews will inform future device iterations
to optimise patient experience.

F-013
Characterization of the Classical Opioid Receptors and Opioid
Growth Factor Receptor in the Human Fetal Membranes. Kaikeline
McCarthy†, Claire E Kendal-Wright*. Chaminade University of Honolulu,
Honolulu, HI, United States.
INTRODUCTION: Opioid use in the United States is a significant
problem, with overdose deaths due to opioid abuse having quadrupled
since 1999. This has led the US government to recognize that there is an
ongoing national opioid epidemic (HHS.gov; CDC.gov). In line with this
trend, the prescription of opioids for pain management during pregnancy
has risen from 18.5% in 2000 to 22.8% in 2007, despite our lack of
understanding of the effects of prenatal exposure to opioids (Yazdy et
al., J Pediatr Genet 2015) and their association with; neonatal abstinence
syndrome, spina bifida and prematurity. Currently no literature exists
regarding the expression and function of opioid receptors in the human
fetal membranes. Although it is known that they function to control
nociception, angiogenesis, barrier disruption and vascular integrity in other
cell types. Therefore, the objective of this study was to characterize the
expression of the various opioid receptors (delta (DOR), kappa (KOR),
and mu (MOR)) and opioid growth factor receptor (OGFR) in the human
fetal membranes.
METHODS: Amniotic mesenchymal cells (AMC), amniotic epithelial
cells (AEC), full thickness fetal membrane (amnion, chorion and decidua),
and amnion-only samples were collected from repeat caesarean section at
Kapi'olani Medical Center for Women and Children with IRB approval.
RNA was isolated from the tissues and cells using Qiagen RNeasy Mini
and Midi Kits (Qiagen, CA, USA). RT-PCR were carried out using HighCapacity RNA-to-cDNA Kit and TaqMan Gene Expression Master Mix
(Applied Biosystems Inc, CA, USA), respectively. Immunohistochemistry
(IHC) was performed on human fetal membrane rolls and pieces of human
placenta with Vectastain Elite ABC-HRP Kit (Vector Laboratories, CA,
USA) according to manufacturer's instructions including a citrate buffer
antigen retrieval step.
RESULTS: The data revealed that DOR, KOR, and OGFR were expressed
at the gene level in the cell types and tissues, at varying levels. OGFR
was the highest expressed receptor, found in AEC, AMC and cells of the
chorion and decidua. DOR was also expressed in all cells tested. KOR was
shown to be expressed in all cells except AMC. However, the expression of

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Table of Contents for the Digital Edition of SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018