Cervical Remodeling Research Articles

Material properties of mouse cervical tissue in normal gestation

An appropriately timed cervical remodeling process is critical for a healthy delivery, yet little is known about the material property changes of the cervix in pregnancy because obtaining human tissue samples is difficult. Rodent models offer advantages including accurately timed pregnant tissues and genetically altered models. Determining the material properties of the mouse cervix, however, is challenging because of its small size and complex geometry. The aim of this study is to quantify cervical material property changes in a normal mouse pregnancy using a microstructurally-inspired porous fiber composite model. We mechanically test intact, whole, gestation-timed mouse cervix by pulling apart tensioned sutures through its inner canal. To interpret our mechanical testing results, we conduct an inverse finite element analysis, taking into account the combined loading state of the thick-walled cylindrical tissue. We fit the material model to previous osmotic swelling data and load-deformation data from this study using a nonlinear optimization scheme, and validate the model by predicting a separate set of deformation data. Overall, the proposed porous fiber composite model captures the mechanical behavior of the mouse cervix in large deformation. The evolution of cervical material parameters indicates that in a normal mouse pregnancy, the cervix begins to soften between day 6 and day 12 of a 19-day gestation period. The material parameter associated with the collagen fiber stiffness decreases from 3.4MPa at gestation day 6 to 9.7e−4MPa at gestation day 18, while the ground substance stiffness decreases from 2.6e−1MPa to 7.0e−4MPa. Statement of SignificanceAccelerated cervical remodeling can lead to extremely premature births. Little is known, however, about the material property changes of the cervix in pregnancy because pregnant human tissue samples are limited. Rodent models overcome this limitation and provide access to gestation-timed samples. Measuring the material property changes of the mouse cervix in pregnancy is challenging due to its small size and complex geometry. Here, we establish a combined experimental and modeling framework. We use this framework to determine the cervical material property changes throughout a normal mouse pregnancy. We present our experimental methods for mechanically testing whole, intact cervical tissue samples. We fit a porous fiber composite material model to the mechanical data and show that the mouse cervix begins to soften between day 6 and day 12 of a 19-day gestation period.

Changes in ectocervical surface area in women throughout pregnancy compared to non-pregnant and postpartum states

Objective: The objective of this study is to estimate changes in the surface area of the ectocervix (CA) in women during pregnancy and compare this to postpartum and non-pregnant states.Methods: CA was evaluated in 210 normal nulliparous women divided into groups from early to late gestation, 40 postpartum women, and 25 non-pregnant women. CA in cm2 was estimated from analysis of images taken with an endoscope of the cervical face and an mm scale. An mm scale was also used to determine fornix length and fornix area computed.Results: The face, fornix, and total areas of the CA of non-pregnant and postpartum groups are significantly smaller (p < 0.001) than these areas in groups during pregnancy. Generally, the CA of the face, fornix, and total area are also less in early pregnancy compared with late gestation (p < 0.01 to <0.001). Total CA correlates with gestational age (r = 0.196, p < 0.004).Conclusions: (1) During pregnancy, CA slowly and progressively increases to >75% area compared with CA of non-pregnant patients and then reverts back to low CA postpartum. (2) Increases in CA during pregnancy occur in both the face and fornix areas. (3) Increases in CA reflect enlargement in cervical volume and remodeling during pregnancy.

Elevated concentration of secretory leukocyte protease inhibitor in the cervical mucus before delivery

Cervical remodeling during parturition progresses under exquisite regulation by immunologic mediators and proteases. Secretory leukocyte protease inhibitor is a secretory protein that can function as an antimicrobial peptide, an antiinflammatory molecule, and aprotease inhibitor. The involvement of secretory leukocyte protease inhibitor in cervical remodeling before and during parturition is understood poorly. We aimed to reveal the role of secretory leukocyte protease inhibitor in the cervical remodeling process before normal term delivery and to evaluate its utility as a predictive biomarker for timing of delivery. Cervical mucus samples were collected prospectively at weekly prenatal visits from a cohort of pregnant women at term. Thesecretory leukocyte protease inhibitor concentrations in 95 mucus samples that were obtained from 49 women with uncomplicated pregnancy who subsequently underwent normal vaginal delivery were assessed. Alterations in secretory leukocyte protease inhibitor concentrations at term and the association of secretory leukocyte protease inhibitor levels with the time to delivery were analyzed. A moderate positive correlation with significance was detected between cervical mucus secretory leukocyte protease inhibitor concentrations and days to delivery (r= 0.38; P= .0001). The secretory leukocyte protease inhibitor concentration was significantly higher in samples that were collected within 7 days of delivery when compared with samples that were collected >7 days before delivery (P= .001). Secretory leukocyte protease inhibitor concentrations were also significantly higher in samples from women with premature rupture of membranes when compared with those without premature rupture of membranes (P= .01), all of whom delivered within 7 days. A logistic regression analysis revealed that the cervical secretory leukocyte protease inhibitor level was a significant parameter for the prediction of the onset of delivery. (P= .017; unit odds ratio, 1.28; 95% confidence interval, 1.07-1.61). A cut-off value of cervical secretory leukocyte protease inhibitor/total protein topredict delivery within 7 days was determined to be 1.62 μg/mg (sensitivity, 0.69; specificity, 0.72) using receiver operating characteristic curve-analysis. Secretory leukocyte protease inhibitor concentrations in the cervical mucus elevate progressively before delivery in uncomplicated term pregnancies. Our findings suggest that cervical secretory leukocyte protease inhibitor is a candidate biomarker for delivery prediction.

201: Human cervical smooth muscle stretch increases matrix metalloproteinase secretion: a new mechanism to explain premature cervical remodeling

201: Human cervical smooth muscle stretch increases matrix metalloproteinase secretion: a new mechanism to explain premature cervical remodeling

Collagen and matrix metalloproteinase-2 and -9 in the ewe cervix during the estrous cycle

The cervical collagen remodeling during the estrous cycle of the ewe was examined. The collagen concentration determined by a hydroxyproline assay and the area occupied by collagen fibers (%C), determined by van Gieson staining, were assessed in the cranial and caudal cervix of Corriedale ewes on Days 1 (n = 6), 6 (n = 5), or 13 (n = 6) after estrous detection (defined as Day 0). In addition, the gelatinase activity by in situ and SDS-PAGE gelatin zymographies and matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9, respectively) expression by immunohistochemistry were determined. The collagen concentration and %C were lowest on Day 1 of the estrous cycle (P < 0.04), when MMP-2 activity was highest (P < 0.006) and the ratio of activated to latent MMP-2 trend to be highest (P = 0.0819). The MMP-2 activity was detected in 73% of the homogenized cervical samples, and its expression was mainly detected in active fibroblasts. By contrast, the MMP-9 activity was detected in 9% of the samples, and its scarce expression was associated with plasmocytes, macrophages, and lymphocytes. Matrix metalloproteinase-2 expression was maximal on Day 1 in the cranial cervix and on Day 13 in the caudal cervix and was lower in the cranial than in the caudal cervix (P < 0.0001). This time-dependent increase in MMP-2 expression that differed between the cranial and caudal cervix may reflect their different physiological roles. The decrease in the collagen content and increase in fibroblast MMP-2 activity in sheep cervix on Day 1 of the estrous cycle suggests that cervical dilation at estrus is due to the occurrence of collagen fiber degradation modulated by changes in periovulatory hormone levels.

Development of an Ultrasonic Method to Detect Cervical Remodeling in Vivo in Full-Term Pregnant Women

The objective of this study was to determine whether estimates of ultrasonic attenuation could detect changes in the cervix associated with medically induced cervical remodeling. Thirty-six full-term pregnant women underwent two transvaginal ultrasonic examinations separated in time by 12 h to determine cervical attenuation, cervical length and changes thereof. Ultrasonic attenuation and cervical length data were acquired from a zone (Zonare Medical Systems, Mountain View, CA, USA) ultrasound system using a 5–9 MHz endovaginal probe. Cervical attenuation and cervical length significantly decreased in the 12 h between the pre-cervical ripening time point and 12 h later. The mean cervical attenuation was 1.1 ± 0.4 dB/cm-MHz before cervical ripening agents were used and 0.8 ± 0.4 dB/cm-MHz 12 h later (p < 0.0001). The mean cervical length also decreased from 3.1 ± 0.9 cm before the cervical ripening was administered to 2.0 ± 1.1 cm 12 h later (p < 0.0001). Cervical attenuation and cervical length detected changes in cervical remodeling 12 h after cervical ripening administration.

Caveolin‐1, a Mechano‐Mediator, is Expressed in the Epithelial and Stromal Cells of Mice Cervix and Increases During Pregnancy

Although we are aware that during pregnancy the developing fetus exerts an increasing gravitational force as it grows in size, little is known about the effect and underlying mechanism of this force on the cervix. Recently our proteomic study provided the first evidence of a presence of a cluster of mechano‐transducing molecules in the cervix, including caveolin‐1 (CAV‐1), talin‐1, vinculin, pro‐filin and others. Here, we use Confocal Immunofluorescence, Western Blot and Real Time PCR analysis to verify our earlier proteomic data by examining the cellular expression, activity (phosphorylated) and levels of CAV‐1 protein and RNA in different stages of pregnancy, non‐pregnant and postpartum (days 0, days 11, 17, postpartum 8, 24 and 48 hours) in mice. CAV‐1 was localized in the epithelial and stromal (fibroblast) cells of the cervix of pregnant mice and its expression increased with advancement in pregnancy but decreased postpartum. The expression was found to be confined to the membrane in late pregnancy and postpartum. We conclude that the growing fetus likely induces mechano‐transduction in the cervix and may influence cervical remodeling and most likely the timing of birth. Funding: Office of Student Research, Appalachian State University.

Evidence for presence of Focal adhesion kinase and enhanced activity in epithelial and stromal cells of mice cervix during pregnancy

We recently provided the first evidence of a presence of a cluster of mechano‐transducing molecules in the cervix using proteomics analysis, including caveolin‐1 (CAV‐1), talin‐1, vinculin, pro‐filin and others. Here, we use confocal immunofluorescence, Western blot and real time PCR analysis to examine whether a key mechano‐transduction molecule, phosphorylated focal adhesion kinase protein and mRNA (pFAK and FAK mRNA) are expressed in the cervix during pregnancy (days 0, days 14, 15, 16), as well as identify its (pFAK) cellular distribution in mice. pFAK was localized in the epithelial and stromal (fibroblast) cells of the cervix of pregnant mice and its expression and activity increased with advancement in pregnancy but decreased postpartum. The expression was found to be confined to the cell membrane. We conclude that the growing fetus likely induces mechano‐transduction in the cervix and may influence cervical remodeling and most likely the timing of birth. Funding: Office of Student Research, Appalachian State University.

A continuous fiber distribution material model for human cervical tissue

The uterine cervix during pregnancy is the vital mechanical barrier which resists compressive and tensile loads generated from a growing fetus. Premature cervical remodeling and softening is hypothesized to result in the shortening of the cervix, which is known to increase a woman׳s risk of preterm birth. To understand the role of cervical material properties in preventing preterm birth, we derive a cervical material model based on previous mechanical, biochemical and histological experiments conducted on nonpregnant and pregnant human hysterectomy cervical tissue samples. In this study we present a three-dimensional fiber composite model that captures the equilibrium material behavior of the tissue in tension and compression. Cervical tissue is modeled as a fibrous composite material, where a single family of preferentially aligned and continuously distributed collagen fibers are embedded in a compressible neo-Hookean ground substance. The total stress in the collagen solid network is calculated by integrating the fiber stresses. The shape of the fiber distribution is described by an ellipsoid where semi-principal axis lengths are fit to optical coherence tomography measurements. The composite material model is fit to averaged mechanical testing data from uni-axial compression and tension experiments, and averaged material parameters are reported for nonpregnant and term pregnant human cervical tissue. The model is then evaluated by investigating the stress and strain state of a uniform thick-walled cylinder under a compressive stress with collagen fibers preferentially aligned in the circumferential direction. This material modeling framework for the equilibrium behavior of human cervical tissue serves as a basis to determine the role of preferentially-aligned cervical collagen fibers in preventing cervical deformation during pregnancy.

Mechanical assessment of cervical remodelling in pregnancy: insight from a synthetic model

During the gestation and the cervical remodelling, several changes occur progressively in the structure of the tissue. An increase in the hydration, disorganisation of collagen network and decrease in elasticity can be observed. The collagen structure disorganisation is particularly complex: collagen fibres turn thicker and more wavy as the gestation progresses in a transition from relatively straight fibres to wavy fibres, while pores between collagen fibres become larger and separated. Shear wave elastography is a promising but not yet fully understood tool to assess these structural changes and the cervix׳s ability to dilate. To this end, a numerical histo-mechanical model is proposed in the present study, which aims at linking variations in the microscopic histo-biomechanical processes with shear wave propagation characteristics. Parametric simulations are carried out for a broad range of mechanical and geometrical parameters. Results show a direct relationship between the histological and morphological changes during pregnancy and the viscoelastic behaviour of the tissue.

816: Nonpregnant women with previous premature cervical remodeling have weaker cervical collagen crosslink maturity ratios

816: Nonpregnant women with previous premature cervical remodeling have weaker cervical collagen crosslink maturity ratios

454: Cervical inflammation and the epithelial barrier: does nitric oxide production lead to cervical remodeling and preterm birth?

454: Cervical inflammation and the epithelial barrier: does nitric oxide production lead to cervical remodeling and preterm birth?

117 IN VIVO EVALUATION OF THE CERVICAL STIFFNESS EVOLUTION DURING INDUCED LABOR IN EWES USING ELASTOGRAPHY

Despite numerous advances and intensive research in perinatal medicine, spontaneous preterm birth (PTB) is the leading global cause of neonatal mortality and morbidity. On the other hand, labour has to be induced in ~23% of pregnancies worldwide. Both issues may be related to the distensibility of the cervical tissue. Quantitative and objective monitoring of the cervix ripening may provide a complementary method to identify cases at risk of PTB and assess the likelihood of successful induction of labour. Currently, however, no reliable clinical tools for such a quantitative and objective evaluation exist. Elastography aims at imaging tissue stiffness. All elastography techniques rely on the same basics: an external force is applied to the tissue and the resulting movements are then followed. Supersonic shear imaging (SSI) is a dynamic method that uses the propagation of mechanical waves to excite the tissue. Its speed is tracked then by ultrafast imaging, allowing characterisation of stiffness [Bercoff et al. 2004 IEEE Trans. Ultrason. Ferroelect. Freq. Contr. 51, 396–409]. Understanding the mechanisms that take place in normal pregnancy will allow a better comprehension of the cervical remodelling and lead to better methods of diagnosis of PTB and successful induction of labour. In this work, we propose a preliminary assessment of the evolution of stiffness during the cervical maturation process in the sheep. The main goal was to study the feasibility of elastography using SSI to quantify cervical stiffness during the maturation process and to assess the potential of this technique for diagnosis of preterm labour and for labour induction success. Cervical stiffness was quantified, by 2 different operators, in 9 pregnant ewes in vivo by using SSI. The cervical ripening was induced by a dexamethasone injection in 5 animals, and 4 animals constituted the control group. The stiffness of the second ring of the cervix was quantified over a circular region of interest of 5 mm of diameter during vaginal ultrasound examination. Images were acquired every 4 h during 24 h to monitor the cervical maturation induced by the dexamethasone injection. Cervical stiffness was found to decrease significantly throughout the cervical ripening (from 9.5 ± 0.9 kPa to 5.0 ± 0.8 kPa; P = 2.7e–5). The intraobserver and interobserver repeatability of measurements were assessed using Bland-Altman analysis with 95% CI. The principal findings of the study were that elastography measurements using SSI technique were highly reproducible in all cases. Second, stiffness of the uterine cervix decreases throughout the maturation process induced by the dexamethasone injection. Finally, it was possible to quantify the decrease of stiffness through the cervical maturation process. Elastography may be a valuable method to quantify objectively and noninvasively the cervical stiffness in vivo, and ultimately could be a useful tool for the diagnosis of PTB and the assessment of labour induction success.

Quantitative evaluation of collagen crosslinks and corresponding tensile mechanical properties in mouse cervical tissue during normal pregnancy.

The changes in the mechanical integrity of the cervix during pregnancy have implications for a successful delivery. Cervical collagens are known to remodel extensively in mice with progressing gestation leading to a soft cervix at term. During this process, mature crosslinked collagens are hypothesized to be replaced with immature less crosslinked collagens to facilitate cervical softening and ripening. To determine the mechanical role of collagen crosslinks during normal mouse cervical remodeling, tensile load-to-break tests were conducted for the following time points: nonpregnant (NP), gestation day (d) 6, 12, 15, 18 and 24 hr postpartum (PP) of the 19-day gestation period. Immature crosslinks (HLNL and DHLNL) and mature crosslinks (DPD and PYD) were measured using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). There were no significant changes in the total immature crosslink density (HLNL+DHLNL mol per collagen mol) throughout normal mouse gestation (range: 0.31–0.49). Total mature crosslink density (PYD+DPD mol per collagen mol) decreased significantly in early softening from d6 to d15 (d6: 0.17, d12: 0.097, d15: 0.026) and did not decrease with further gestation. The maturity ratio (total mature to total immature crosslinks) significantly decreased in early softening from d6 to d15 (d6: 0.2, d15: 0.074). All of the measured crosslinks correlated significantly with a measure of tissue stiffness and strength, with the exception of the immature crosslink HLNL. This data provides quantitative evidence to support the hypothesis that as mature crosslinked collagens decline, they are replaced by immature collagens to facilitate increased tissue compliance in the early softening period from d6 to d15.

Analysis of miRNA–mRNA expression profiles in decidual natural killer cells during early pregnancy

s for the 29th Annual Congress of Japanese Society / Journal of Reproductive Immunology 106 (2014) 1–20 17 LPS stimulation induced a significant increase of transcript levels of matrix metalloproteinase (MMP)-1, MMP-3, and MMP-9, while P4 treatment significantly attenuated LPSinduced increase of transcript levels of these MMPs. Conclusions: Our preliminary results suggest that P4 might prevent spontaneous preterm labor by inhibiting cervical remodeling, particularly through the attenuation of ECM degradation by inhibiting MMPs production. http://dx.doi.org/10.1016/j.jri.2014.09.040 Analysis of miRNA–mRNA expression profiles in decidual natural killer cells during early pregnancy D. Zhao1,∗, B. Than Naing1, K. Inada2, T. Shima2, S. Saito2, T. Takeshita3, T. Takizawa1 1 Department of Molecular Medicine & Anatomy, Nippon Medical School, Tokyo, Japan 2 Department of Obstetrics and Gynecology, University of Toyama, Toyama, Japan 3 Department of Obstetrics and Gynecology, Nippon Medical School, Tokyo, Japan Objective: Decidual natural killer (dNK) cells play an important role in the maintenance of early pregnancy. However, the expression profiles and functions of miRNAs within dNK cells remain to be clarified.We investigated the miRNA–mRNAexpressionprofiles in dNK cells during early pregnancy. Methods:Decidual tissue and peripheral blood samples from patients who gave informed consent were aseptically obtained after legal abortions (at 6–7 weeks of gestation, n=3). We compared miRNA-mRNA expression patterns between dNK and peripheral NK (pNK) cells. The expression levels of miRNAs were examined by real-time PCR using a TaqMan microRNA Assay, and gene expression profiling was conducted using an Agilent microarray. Networks, functions, and pathways analyses were generated using Ingenuity Pathway Analysis (IPA). Results and conclusions: Among the 750 miRNAs examined, 199 miRNAs were significantly up-regulated in dNK cells compared with pNK cells. We detected some placenta-specific miRNAs derived from the chromosome 19 microRNA cluster in dNK cells. Among the 34,127 transcripts examined, 534 genes were significantly downregulated in dNK cells compared with pNK cells; the down-regulated gene expression profile exhibited low cytotoxicity in dNK cells. Some of the up-regulated miRNAs in dNK cells were the miRNAs potentially targeting the above cytotoxicity-associated genes. These data suggest that dNKcells have auniquemiRNA-mRNA interaction network for the maintenance of early pregnancy. http://dx.doi.org/10.1016/j.jri.2014.09.041 Frequent Chlamydophila pneumoniae infection in vertically HIV-infected children in Vietnam Q.D. Trinh ∗, N.T. Pham, N.T. Le Nguyen, B.Q. Lam, K.T. Le Phan, K.H. Truong, T.Q. Le, H.T. Nguyen, T.C. Tang, Y. Izumi, S. Komine-Aizawa, H. Ushijima, S. Hayakawa Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Japan Objective: Chlamydophila pneumoniae (C. pneumoniae) is an intracellular pathogen and often cause fatal respiratory symptoms in HIV/AIDS patients. However, its frequency in children with vertical HIV infection is so far unknown. Methods: A total of 89 blood samples collected from HIV-infected infants and children from provinces of southern Vietnam who were hospitalized at Children’s Hospital 1, Ho Chi Minh City, during the 1-year period fromOctober 2004 to September 2005 were submitted to serological screening for IgG, IgA, and IgM antibodies against C. pneumoniae. The presence of this microorganism was also evaluated by PCR. Age matched control enrolled HIV negative 91 children with other infectious disorders. Results: The results showed that 64% of the samples were positive for anti-C. pneumoniae IgG, 31.5% were positive for IgA, and 3.4% were positive for IgM. The highest prevalences of IgG and IgA positivity, 75% and 66.7%, respectively, were noted in the 1to 2-year-old age group. In control patients, IgG and IgA positivitywere 7.7 and 4.4% respectively. No patients showed positive PCR results. Conclusions: Our study revealed a high seroprevalence of C. pneumoniae in Vietnamese infants and children with HIV/AIDS by vertical transmission. http://dx.doi.org/10.1016/j.jri.2014.09.042 Porphyromonas gingivalis inhibits trophoblast invasion N. Hirohata ∗, S. Komine-Aizawa, S. Hayakawa Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Japan Objective: Periodontitis has been regarded as a risk factor of pregnancy complications including preterm birth, intrauterine growth retardation and preeclampsia. However protective roles of dental intervention during pregnancyon fetal/maternal outcomes are controversial. In this study, we investigated in vitro effects of a periodontal bacteria Porphyromonas gingivalis on cultured trophoblasts and mucosal natural killer cells to mimic in vivo phenomena at the feto-maternal interface. Methods: 2×109 of P. gingivalis (ATCC33277)were cultured at 37 ◦C, 10% CO2 in BHIB media supplemented with 0.005mgml−1 hemin and 0.0005mgml−1 menadione for

Progesterone transcriptionally inhibits LPS-induced matrix metalloproteinase up-regulation in human cervical fibroblast cells

s for the 29th Annual Congress of Japanese Society / Journal of Reproductive Immunology 106 (2014) 1–20 17 LPS stimulation induced a significant increase of transcript levels of matrix metalloproteinase (MMP)-1, MMP-3, and MMP-9, while P4 treatment significantly attenuated LPSinduced increase of transcript levels of these MMPs. Conclusions: Our preliminary results suggest that P4 might prevent spontaneous preterm labor by inhibiting cervical remodeling, particularly through the attenuation of ECM degradation by inhibiting MMPs production. http://dx.doi.org/10.1016/j.jri.2014.09.040 Analysis of miRNA–mRNA expression profiles in decidual natural killer cells during early pregnancy D. Zhao1,∗, B. Than Naing1, K. Inada2, T. Shima2, S. Saito2, T. Takeshita3, T. Takizawa1 1 Department of Molecular Medicine & Anatomy, Nippon Medical School, Tokyo, Japan 2 Department of Obstetrics and Gynecology, University of Toyama, Toyama, Japan 3 Department of Obstetrics and Gynecology, Nippon Medical School, Tokyo, Japan Objective: Decidual natural killer (dNK) cells play an important role in the maintenance of early pregnancy. However, the expression profiles and functions of miRNAs within dNK cells remain to be clarified.We investigated the miRNA–mRNAexpressionprofiles in dNK cells during early pregnancy. Methods:Decidual tissue and peripheral blood samples from patients who gave informed consent were aseptically obtained after legal abortions (at 6–7 weeks of gestation, n=3). We compared miRNA-mRNA expression patterns between dNK and peripheral NK (pNK) cells. The expression levels of miRNAs were examined by real-time PCR using a TaqMan microRNA Assay, and gene expression profiling was conducted using an Agilent microarray. Networks, functions, and pathways analyses were generated using Ingenuity Pathway Analysis (IPA). Results and conclusions: Among the 750 miRNAs examined, 199 miRNAs were significantly up-regulated in dNK cells compared with pNK cells. We detected some placenta-specific miRNAs derived from the chromosome 19 microRNA cluster in dNK cells. Among the 34,127 transcripts examined, 534 genes were significantly downregulated in dNK cells compared with pNK cells; the down-regulated gene expression profile exhibited low cytotoxicity in dNK cells. Some of the up-regulated miRNAs in dNK cells were the miRNAs potentially targeting the above cytotoxicity-associated genes. These data suggest that dNKcells have auniquemiRNA-mRNA interaction network for the maintenance of early pregnancy. http://dx.doi.org/10.1016/j.jri.2014.09.041 Frequent Chlamydophila pneumoniae infection in vertically HIV-infected children in Vietnam Q.D. Trinh ∗, N.T. Pham, N.T. Le Nguyen, B.Q. Lam, K.T. Le Phan, K.H. Truong, T.Q. Le, H.T. Nguyen, T.C. Tang, Y. Izumi, S. Komine-Aizawa, H. Ushijima, S. Hayakawa Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Japan Objective: Chlamydophila pneumoniae (C. pneumoniae) is an intracellular pathogen and often cause fatal respiratory symptoms in HIV/AIDS patients. However, its frequency in children with vertical HIV infection is so far unknown. Methods: A total of 89 blood samples collected from HIV-infected infants and children from provinces of southern Vietnam who were hospitalized at Children’s Hospital 1, Ho Chi Minh City, during the 1-year period fromOctober 2004 to September 2005 were submitted to serological screening for IgG, IgA, and IgM antibodies against C. pneumoniae. The presence of this microorganism was also evaluated by PCR. Age matched control enrolled HIV negative 91 children with other infectious disorders. Results: The results showed that 64% of the samples were positive for anti-C. pneumoniae IgG, 31.5% were positive for IgA, and 3.4% were positive for IgM. The highest prevalences of IgG and IgA positivity, 75% and 66.7%, respectively, were noted in the 1to 2-year-old age group. In control patients, IgG and IgA positivitywere 7.7 and 4.4% respectively. No patients showed positive PCR results. Conclusions: Our study revealed a high seroprevalence of C. pneumoniae in Vietnamese infants and children with HIV/AIDS by vertical transmission. http://dx.doi.org/10.1016/j.jri.2014.09.042 Porphyromonas gingivalis inhibits trophoblast invasion N. Hirohata ∗, S. Komine-Aizawa, S. Hayakawa Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Japan Objective: Periodontitis has been regarded as a risk factor of pregnancy complications including preterm birth, intrauterine growth retardation and preeclampsia. However protective roles of dental intervention during pregnancyon fetal/maternal outcomes are controversial. In this study, we investigated in vitro effects of a periodontal bacteria Porphyromonas gingivalis on cultured trophoblasts and mucosal natural killer cells to mimic in vivo phenomena at the feto-maternal interface. Methods: 2×109 of P. gingivalis (ATCC33277)were cultured at 37 ◦C, 10% CO2 in BHIB media supplemented with 0.005mgml−1 hemin and 0.0005mgml−1 menadione for

Frequent Chlamydophila pneumoniae infection in vertically HIV-infected children in Vietnam

s for the 29th Annual Congress of Japanese Society / Journal of Reproductive Immunology 106 (2014) 1–20 17 LPS stimulation induced a significant increase of transcript levels of matrix metalloproteinase (MMP)-1, MMP-3, and MMP-9, while P4 treatment significantly attenuated LPSinduced increase of transcript levels of these MMPs. Conclusions: Our preliminary results suggest that P4 might prevent spontaneous preterm labor by inhibiting cervical remodeling, particularly through the attenuation of ECM degradation by inhibiting MMPs production. http://dx.doi.org/10.1016/j.jri.2014.09.040 Analysis of miRNA–mRNA expression profiles in decidual natural killer cells during early pregnancy D. Zhao1,∗, B. Than Naing1, K. Inada2, T. Shima2, S. Saito2, T. Takeshita3, T. Takizawa1 1 Department of Molecular Medicine & Anatomy, Nippon Medical School, Tokyo, Japan 2 Department of Obstetrics and Gynecology, University of Toyama, Toyama, Japan 3 Department of Obstetrics and Gynecology, Nippon Medical School, Tokyo, Japan Objective: Decidual natural killer (dNK) cells play an important role in the maintenance of early pregnancy. However, the expression profiles and functions of miRNAs within dNK cells remain to be clarified.We investigated the miRNA–mRNAexpressionprofiles in dNK cells during early pregnancy. Methods:Decidual tissue and peripheral blood samples from patients who gave informed consent were aseptically obtained after legal abortions (at 6–7 weeks of gestation, n=3). We compared miRNA-mRNA expression patterns between dNK and peripheral NK (pNK) cells. The expression levels of miRNAs were examined by real-time PCR using a TaqMan microRNA Assay, and gene expression profiling was conducted using an Agilent microarray. Networks, functions, and pathways analyses were generated using Ingenuity Pathway Analysis (IPA). Results and conclusions: Among the 750 miRNAs examined, 199 miRNAs were significantly up-regulated in dNK cells compared with pNK cells. We detected some placenta-specific miRNAs derived from the chromosome 19 microRNA cluster in dNK cells. Among the 34,127 transcripts examined, 534 genes were significantly downregulated in dNK cells compared with pNK cells; the down-regulated gene expression profile exhibited low cytotoxicity in dNK cells. Some of the up-regulated miRNAs in dNK cells were the miRNAs potentially targeting the above cytotoxicity-associated genes. These data suggest that dNKcells have auniquemiRNA-mRNA interaction network for the maintenance of early pregnancy. http://dx.doi.org/10.1016/j.jri.2014.09.041 Frequent Chlamydophila pneumoniae infection in vertically HIV-infected children in Vietnam Q.D. Trinh ∗, N.T. Pham, N.T. Le Nguyen, B.Q. Lam, K.T. Le Phan, K.H. Truong, T.Q. Le, H.T. Nguyen, T.C. Tang, Y. Izumi, S. Komine-Aizawa, H. Ushijima, S. Hayakawa Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Japan Objective: Chlamydophila pneumoniae (C. pneumoniae) is an intracellular pathogen and often cause fatal respiratory symptoms in HIV/AIDS patients. However, its frequency in children with vertical HIV infection is so far unknown. Methods: A total of 89 blood samples collected from HIV-infected infants and children from provinces of southern Vietnam who were hospitalized at Children’s Hospital 1, Ho Chi Minh City, during the 1-year period fromOctober 2004 to September 2005 were submitted to serological screening for IgG, IgA, and IgM antibodies against C. pneumoniae. The presence of this microorganism was also evaluated by PCR. Age matched control enrolled HIV negative 91 children with other infectious disorders. Results: The results showed that 64% of the samples were positive for anti-C. pneumoniae IgG, 31.5% were positive for IgA, and 3.4% were positive for IgM. The highest prevalences of IgG and IgA positivity, 75% and 66.7%, respectively, were noted in the 1to 2-year-old age group. In control patients, IgG and IgA positivitywere 7.7 and 4.4% respectively. No patients showed positive PCR results. Conclusions: Our study revealed a high seroprevalence of C. pneumoniae in Vietnamese infants and children with HIV/AIDS by vertical transmission. http://dx.doi.org/10.1016/j.jri.2014.09.042 Porphyromonas gingivalis inhibits trophoblast invasion N. Hirohata ∗, S. Komine-Aizawa, S. Hayakawa Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Japan Objective: Periodontitis has been regarded as a risk factor of pregnancy complications including preterm birth, intrauterine growth retardation and preeclampsia. However protective roles of dental intervention during pregnancyon fetal/maternal outcomes are controversial. In this study, we investigated in vitro effects of a periodontal bacteria Porphyromonas gingivalis on cultured trophoblasts and mucosal natural killer cells to mimic in vivo phenomena at the feto-maternal interface. Methods: 2×109 of P. gingivalis (ATCC33277)were cultured at 37 ◦C, 10% CO2 in BHIB media supplemented with 0.005mgml−1 hemin and 0.0005mgml−1 menadione for

Role of Increasing Vaginal Acidity in Histological Changes and Softening of the Cervix Prior to Dilatation in Perimenopausal Women

Background: cervical dilatation, along with hysteroscopy or endometrial curettage, is one of the most common procedures in gynecological practice. However, difficulty in entering the internal cervical os may be encountered; Therefore, there is a need for an effective cervical priming agent which can be used in general gynecological practice. Vaginal acidification might increase collagen degradation. It is known that cervical collagen degradation contributes to the ripening process. Aim of the work: the present study was designed to evaluate the efficacy of acidifying vaginal cream in cervical tissue remodeling and softening. Patients and methods: this study is a double-blinded clinical trial study, conducted in Obstetrics and Gynecology Department of Al-Azhar University hospital (Damietta). It included fifty perimenopausal women, their age ranged from 45-55ys, planned for cervical dilatation before endometrial biopsy due to irregular uterine bleeding. They were divided into two main groups: Group I: women who were treated with acidifying vaginal cream (Eva base cream melted with acetic acid 5%). Group II: women who were treated with neutral (placebo) vaginal cream (Eva base cream only). Efficacy of acidifying vaginal cream on the cervix was assessed on the basis of improved cervical softening and degree of cervical changes on the histological examination. Results: there was no significant difference in both groups during the preoperative assessment of the cervix and premedication. During operation in the study group (I) 1st Hegar’s passed was No8 in 52% of cases and No9 in 28% of cases denoting the significant effect of acetic acid on cervical dilation in comparison with placebo group (II)in which the cervix was closed in 44% of cases and Hegar’s No 3 passed in 36% of cases showing statistical significance between the two groups. As regards the histological examination, it was found that in group (I) collagen dissociation grade II represent 56% and grade III 24% with statistical significance in comparison with placebo group (II) in which collagen was grade 0, in 76% . Also, the inflammatory changes in group(I) showed moderate changes in 72% of cases and 20% showed severe changes with statistical significance in comparison with the control group(II) in which 56% of cases showed no inflammatory changes. Conclusion: vaginal acidification with 5% acetic acid as a cervical softening agent appeared to be safe, effective, with no adverse effects.

Raman spectroscopy provides a noninvasive approach for determining biochemical composition of the pregnant cervix in vivo

The molecular changes that occur with cervical remodelling during pregnancy are not completely understood. This study reviews Raman spectroscopy, an optical technique for detecting changes in the pregnant cervix, and reports preliminary studies on cervical remodelling in mice that suggest that the technique provides advantages over other methods. Raman spectroscopy is sensitive to biochemical changes in the pregnant cervix and has high potential as a tool for detecting premature cervical remodelling in pregnant women.

Androgens in pregnancy: roles in parturition

BACKGROUNDUnderstanding the physiology of pregnancy enables effective management of pregnancy complications that could otherwise be life threatening for both mother and fetus. A functional uterus (i) retains the fetus in utero during pregnancy without initiating stretch-induced contractions and (ii) is able to dilate the cervix and contract the myometrium at term to deliver the fetus. The onset of labour is associated with successful cervical remodelling and contraction of myometrium, arising from concomitant activation of uterine immune and endocrine systems. A large body of evidence suggests that actions of local steroid hormones may drive changes occurring in the uterine microenvironment at term. Although there have been a number of studies considering the potential role(s) played by progesterone and estrogen at the time of parturition, the bio-availability and effects of androgens during pregnancy have received less scrutiny. The aim of this review is to highlight potential roles of androgens in the biology of pregnancy and parturition.METHODSA review of published literature was performed to address (i) androgen concentrations, including biosynthesis and clearance, in maternal and fetal compartments throughout gestation, (ii) associations of androgen concentrations with adverse pregnancy outcomes, (iii) the role of androgens in the physiology of cervical remodelling and finally (iv) the role of androgens in the physiology of myometrial function including any impact on contractility.RESULTSSome, but not all, androgens increase throughout gestation in maternal circulation. The effects of this increase are not fully understood; however, evidence suggests that increased androgens might regulate key processes during pregnancy and parturition. For example, androgens are believed to be critical for cervical remodelling at term, in particular cervical ripening, via regulation of cervical collagen fibril organization. Additionally, a number of studies highlight potential roles for androgens in myometrial relaxation via non-genomic, AR-independent pathways critical for the pregnancy reaching term. Understanding of the molecular events leading to myometrial relaxation is an important step towards development of novel targeted tocolytic drugs.CONCLUSIONSThe increase in androgen levels throughout gestation is likely to be important for establishment and maintenance of pregnancy and initiation of parturition. Further investigation of the underlying mechanisms of androgen action on cervical remodelling and myometrial contractility is needed. The insights gained may facilitate the development of new therapeutic approaches to manage pregnancy complications such as preterm birth.