Uterine Segments Research Articles

Expression Analysis of Prostaglandin E and F Receptors in Rat Uterus, Mesenteric Artery and Uterine Artery for Development of a Surrogate Assay of Uterine Contractility

Preterm labor occurs in nearly 10% of U.S. pregnancies and is the leading cause of neonatal morbidity and mortality worldwide. Prostaglandins, specifically of the E and F series, play a key role in myometrial contractility and have been recognized as activators of parturition. As such, modulation of select prostaglandin signaling pathways provides a potential approach to pharmaceutical intervention of preterm labor. Prostaglandin E2 is the most abundantly produced prostanoid and functions through four subtypes of G protein coupled receptors (EP1, EP2, EP3, and EP4). It is generally accepted that EP1 and EP3 serve as pro‐contractility receptors while EP2 and EP4 are relaxatory receptors that promote myometrial quiescence. Prostaglandin F2α (PGF2α) functions through the FP receptor and plays an essential role in uterine contractility during preterm or term labor and delivery. The dynamicity and complexity of prostaglandin pharmacology is an important factor to consider during assay development for the testing of molecules which may not be selective to one receptor subtype.Translational ex vivo assays that enhance throughput of drug candidate screening and decrease the use of experimental animals are increasingly desirable. The objective of this research was to determine the relevance of employing either mesenteric resistance artery or uterine artery segments as surrogate tissues to the uterus in the evaluation of prostaglandin‐induced smooth muscle contraction. The relative expression of EP1, EP2, EP3, EP4, and FP receptor mRNA was determined by RT‐qPCR and compared between the uterine body, uterine artery, and mesenteric artery in the rat. Additionally, the efficacy of PGF2α in eliciting contraction was measured by isometric wire myography and compared between the uterine and mesenteric artery.The results of this study demonstrate that rat mesenteric artery may have utility as a model in the screening of prostaglandin receptor‐targeting molecules. Together, these findings will contribute to the identification of suitable and efficient ex vivo assays by which new pharmacological approaches for the treatment of preterm labor can be evaluated.Support or Funding InformationFerring Research Institute, San Diego, CAThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Adaptive reduction of human myometrium contractile activity in response to prolonged uterine stretch during term and twin pregnancy. Role of TREK-1 channel

Quiescence of myometrium contractile activity allows uterine expansion to accommodate the growing fetus and prevents preterm labor particularly during excessive uterine stretch in multiple pregnancy. However, the mechanisms regulating uterine response to stretch are unclear. We tested the hypothesis that prolonged uterine stretch is associated with decreased myometrium contractile activity via activation of TWIK-related K+ channel (TREK-1). Pregnant women at different gestational age (preterm and term) and uterine stretch (singleton and twin pregnancy) were studied, and uterine strips were isolated for measurement of contractile activity and TREK-1 channel expression/activity. Both oxytocin- and KCl-induced contraction were reduced in term vs preterm pregnancy and in twin vs singleton pregnancy. Oxytocin contraction was reduced in uterine segments exposed to 8 g stretch compared to control tissues under 2 g basal tension. TREK-1 mRNA expression and protein levels were augmented in Singleton-Term vs Singleton-Preterm, and in uterine strips exposed to 8 g stretch. The TREK-1 activator arachidonic acid reduced oxytocin contraction in preterm and term, singleton and twin pregnant uterus. The TREK-1 blocker l-methionine enhanced oxytocin contraction in Singleton-Term and twin pregnant uterus, and reversed the decreases in contraction in uterine strips exposed to prolonged stretch. Carboprost-induced uterine contraction was also reduced by arachidonic acid and enhanced by l-methionine. Thus, myometrium contraction decreases with gestational age and uterine expansion in twin pregnancy. The results suggest that prolonged stretch enhances the expression/activity of TREK-1 channel, leading to decreased myometrium contractile activity and maintained healthy term pregnancy particularly in multiple pregnancy.

Acute intravenous exposure to silver nanoparticles during pregnancy induces particle size and vehicle dependent changes in vascular tissue contractility in Sprague Dawley rats

The use of silver nanoparticles (AgNP) raises safety concerns during susceptible life stages such as pregnancy. We hypothesized that acute intravenous exposure to AgNP during late stages of pregnancy will increase vascular tissue contractility, potentially contributing to alterations in fetal growth. Sprague Dawley rats were exposed to a single dose of PVP or Citrate stabilized 20 or 110nm AgNP (700μg/kg). Differential vascular responses and EC50 values were observed in myographic studies in uterine, mesenteric arteries and thoracic aortic segments, 24h post-exposure. Reciprocal responses were observed in aortic and uterine vessels following PVP stabilized AgNP with an increased force of contraction in uterine artery and increased relaxation responses in aorta. Citrate stabilized AgNP exposure increased contractile force in both uterine and aortic vessels. Intravenous AgNP exposure during pregnancy displayed particle size and vehicle dependent moderate changes in vascular tissue contractility, potentially influencing fetal blood supply.

Lignocaine augments the in-vitro uterine contractions involving NO-guanylyl cyclase dependent mechanisms

AimsLignocaine is used during intrapartum and postpartum period but there are conflicting reports regarding the effect of lignocaine on uterine contractility. Therefore, this study was undertaken to delineate the effect of lignocaine on uterine contractility and the underlying mechanisms. Main methodsThe in vitro contractions were recorded from the uterine segments obtained from adult rats (in estrous phase) and also from human myometrial tissue. Effect of lignocaine on spontaneous uterine contractions was recorded in the absence or presence of antagonists. Effect of sodium nitroprusside (SNP, NO donor) on uterine contractility was assessed. The NO2− was assayed (indicator of NO activity) from the supernatant after exposing the myometrial tissue to lignocaine in the absence or the presence of L-NAME or hemoglobin. Key findingsLignocaine (100μM) increased the amplitude of uterine contractions by 75% with no alterations in frequency. Similar magnitude of increase was seen with human myometrial tissue also. The spontaneous activities were absent in Ca2+-free or in nifedipine (10μM) containing medium. Heparin (IP3 blocker, 10IU/ml), but not the indomethacin (10μM) blocked the lignocaine-induced augmentation. L-NAME (NOS inhibitor, 10μM) or methylene blue (guanylyl cyclase inhibitor, 100μM) partially blocked the lignocaine-induced augmentation. SNP (30μM) increased the amplitude of spontaneous uterine contractions. Lignocaine increased the NO2− content (indicator of NO activity) of uterine tissue and the increase was blocked by L-NAME or hemoglobin. SignificancePresent observations indicate that lignocaine augments the amplitude of uterine contractions via Ca2+-dependent mechanisms involving NO-G cyclase-dependent mechanisms.

EP19.05: The dynamic changes in upper and lower uterine segment in 3rd stage of labour and blood loss during 3rd stage of labour

The myometrial contraction is an important mechanism to reduce blood loss in 3rd and 4th stage of labour. In this study we evaluated the effect of the dynamic changes in different uterine segments on blood loss in the 3rd stage of labour. In a cohort study anterior (AUS) and posterior upper (PUS) and lower (ALS), (PLS) uterine myometrium thickness were measured in the mid region of the upper and lower uterine segments at one minute time intervals by ultrasound in the 3rd stage of labour in 80 term pregnancies and the thickness changes was compared with the amount of blood loss during 3rd stage of labour. The mean gestational age was 39.17 +/- 0.79 weeks. The mean duration of 3rd stage was 5.65 +/- 2.23 minutes. The mean blood loss in 3rd stage of labour was 371.03 +/- 659.84. The mean hemoglobin concentration was 12.32 +/- 0.97 g/dl and 10.98 +/- 1.44 g/dl before and after delivery respectively. In nulliparas the amount of blood loss in the 3rd stage of labour was significantly more than multiparas (P=0.01). The mean myometrial thickness in AUS increased linearly while in ALS decreased linearly during the process of placental expulsion. There was no significant difference in AUS, PUS, ALS and PLS dynamic changes between nulliparas and multiparas. There was no linear correlation between dynamic changes in different uterine segments and blood loss in the 3rd stage of labour. (AUS: R=0.13, P=0.23, PUS: R=0.03, P=0.73, ALS: R=0.01, P=0.92,PLS: R=0.14, P=0.23) There was a linear increase in AUS thickness and a linear decrease in ALS in 3rd stage of labour which enhanced placental expulsion but had no linear correlation with amount of blood loss.

The effect of anatomical placental location on the third stage of labor.

According to the lower uterine segment and trophotropism hypotheses of placental implantation, placentation occurs in uterine segments with a high blood supply. The present study was designed to determine the clinical implications of these hypotheses by assessing the dynamics of the third stage of labor. Two-hundred and ninety-two prospectively followed pregnant women were assessed at the time of delivery. The uterine locations of the placentas were determined according to the position of the main placental volume and mass under a sagittal view obtained using a centrally located ultrasonography probe. Depending on the location of the placental mass, the patients were assigned to three groups: anterior, posterior, and fundal. The placenta was located in the anterior, posterior, and fundal wall in 52.7% (154/292), 37.7% (110/292), and 9.6% (28/292) of cases, respectively. The duration of the third stage of labor was 6.26 ± 3.89, 6.47 ± 3.68, and 7.42 ± 3.67 min in the anterior, posterior, and fundal placental groups, respectively (P = 0.06). The placenta was separated peripherally in 25% (n = 73) of the patients in each group and centrally in 75% (n = 219). The delta hematocrit value (hematocrit change) was 2.99 ± 4.39, 3.15 ± 6.52, and 2.76 ± 3.04 in the anterior, posterior, and fundal groups, respectively (P = 0.99). This study showed that the location of the placenta did not affect the dynamics of the third stage of labor. Clin. Anat. 30:508-511, 2017. © 2017 Wiley Periodicals, Inc.

Histomorphometrics and quantitative unbiased stereology in canine uteri treated with medroxyprogesterone acetate

This article describes the effects of MPA use on the canine uterus using stereological methods. Entire reproductive tracts were removed from normal healthy canine bitches (Canis lupus familiaris) and grouped as: nulliparous (n = 11), multiparous (n = 11) and MPA-treated (n = 11; nulliparous; two treatments; 5 mg/kg). 1 cm samples were cut from the corpus, horn and uterine tube and fixed in 10% formaldehyde. Sections of each were mounted on slides and stained with hematoxylin-eosin. We assessed the fraction area for components of endometrium and myometrium and VV (volume density) and SV (surface density) of the gland and stroma using the M36 test system provided by the STEPanizer Stereological Tool. No gross histological differences were observed between study groups in the uterine tube, uterine corpus and horn. The wall of the uterine corpus and horn in MPA-treated bitches was characterized as being thicker than in the other groups. A cross-section of the uterine corpus revealed no differences between components of uterine wall in the corpus and horn; however, differences were observed in the volume density [VV; %] in variables such as: VV[str.vasc/uterus] (nulliparous vs. multiparous; p = 0.0019) and VV[str.supravasc/uterus] (multiparous vs. nulliparous and MPA; p = 0.0035). In the endometrial gland, differences were detected in SV[gland/endom] (multiparous vs. MPA, p = 0.0442). In the uterine horn, differences were only observed in the variable VV[lumen.gland/endom] (multiparous vs. MPA; p = 0.0019). This study shows quantitative changes in the architecture of the endometrium and myometrium in all the uterine segments, mainly morphological endometrial gland changes of the uterine corpus, increasing the surface area per unit of volume; however, these changes usually do not differ quantitatively from those observed in the uterus of multiparous bitches.

Expression of vascular endothelial growth factor (VEGF) and factor VIII in the gilt placenta and its relation to fetal development

Vascular endothelial growth factor (VEGF) and von Willebrand factor (Factor VIII) are important components involved in the regulation of vascular development and identification of endothelial cells in many tissues. This study aimed to evaluate the presence of these substances in the placenta of pig fetuses located in different uterine regions and at different gestational ages and correlate them with fetal development. One hundred seventy-five pig fetuses from fifteen gilts slaughtered at 50, 80 and 106 days of pregnancy were used. Each uterine horn was divided into three segments, the apex, base and middle region, and also into left and right sides. The fetuses were sexed before determining their weight and anatomical measurements. The weights of the placentas were obtained for the calculation of placental efficiency, and VEGF and factor VIII were determined by immunohistochemistry. There was no significant interaction between gestational age, uterine segment or side and fetal sex in any of the variables studied. Higher VEGF and factor VIII concentrations were found at 80 and 105 days of pregnancy, and there was no significant difference between the right and left sides of the uterus, uterine segments or fetal sex. Positive correlations between VEGF and fetal weights were observed at 80 and 105 days of pregnancy, whereas factor VIII showed positive correlations with the weight and length of fetuses and placental weight and efficiency throughout pregnancy. It was concluded that VEGF and factor VIII are important growth factors associated with fetal development in pigs and are identified in all uterine segments. The concentration of these substances increases until the middle third of pregnancy which suggests that most of the uterine vascular development occurs before this stage.

Identifying specific proteins involved in eggshell membrane formation using gene expression analysis and bioinformatics

BackgroundThe avian eggshell membranes surround the egg white and provide a structural foundation for calcification of the eggshell which is essential for avian reproduction; moreover, it is also a natural biomaterial with many potential industrial and biomedical applications. Due to the insoluble and stable nature of the eggshell membrane fibres, their formation and protein constituents remain poorly characterized. The purpose of this study was to identify genes encoding eggshell membrane proteins, particularly those responsible for its structural features, by analyzing the transcriptome of the white isthmus segment of the oviduct, which is the specialized region responsible for the fabrication of the membrane fibres.ResultsThe Del-Mar 14 K chicken microarray was used to investigate up-regulated expression of transcripts in the white isthmus (WI) compared with the adjacent magnum (Ma) and uterine (Ut) segments of the hen oviduct. Analysis revealed 135 clones hybridizing to over-expressed transcripts (WI/Ma + WI/Ut), and corresponding to 107 NCBI annotated non-redundant Gallus gallus gene IDs. This combined analysis revealed that the structural proteins highly over-expressed in the white isthmus include collagen X (COL10A1), fibrillin-1 (FBN1) and cysteine rich eggshell membrane protein (CREMP). These results validate previous proteomics studies which have identified collagen X (α-1) and CREMP in soluble eggshell extracts. Genes encoding collagen-processing enzymes such as lysyl oxidase homologs 1, 2 and 3 (LOXL1, LOXL2 and LOXL3), prolyl 4 hydroxylase subunit α-2 and beta polypeptide (P4HA2 and P4HB) as well as peptidyl-prolyl cis-trans isomerase C (PPIC) were also over-expressed. Additionally, genes encoding proteins known to regulate disulfide cross-linking, including sulfhydryl oxidase (QSOX1) and thioredoxin (TXN), were identified which suggests that coordinated up-regulation of genes in the white isthmus is associated with eggshell membrane fibre formation.ConclusionsThe present study has identified genes associated with the processing of collagen, other structural proteins, and disulfide-mediated cross-linking during eggshell membrane formation in the white isthmus. Identification of these genes will provide new insight into eggshell membrane structure and mechanisms of formation that will assist in the development of selection strategies to improve eggshell quality and food safety of the table egg.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-2013-3) contains supplementary material, which is available to authorized users.

Ultrasonographic wall thickness measurement of the upper and lower uterine segments in the prediction of the progress of preterm labour

To assess the role of ultrasonographic measurement of the upper and lower uterine segments wall thickness in predicting the progress of preterm labour in patients presenting with preterm labour pains.Fifty pregnant women presenting at Obstetrics Department - Suez Canal University, Egypt with regular lower abdominal pains and diagnosed as having preterm labour were enrolled in the study.Measurements of the upper and lower uterine segments wall thickness by transabdominal ultrasonography in-between contractions and with full bladder were taken. The upper/lower uterine wall thickness ratio was calculated and correlated to the progress of the preterm labour and to the response to tocolytics.The ultrasonographic upper/lower uterine wall thickness ratio was directly related to the progress of preterm delivery (PTD). The change in this ratio is correlated inversely with the response to tocolysis. Using the ROC curve, when the upper/lower uterine wall thickness ratio was ≤ 1.26 the sensitivity was 94.74 and the specificity was 100.00, and when the ratio was ≤ 1.52 the sensitivity was 100.00 and the specificity was 83.33.These data may serve as a baseline ultrasonographic reference values for further studies in prediction the progress of preterm labour in patients presenting with preterm labour pains.

PGF2α modulates the output of chemokines and pro-inflammatory cytokines in myometrial cells from term pregnant women through divergent signaling pathways.

Prostaglandin F2α (PGF2α) plays a critical role in the initiation and process of parturition. Since human labor has been described as an inflammatory event, we investigated the role of PGF2α in the inflammatory process using cultured human uterine smooth muscle cells (HUSMCs) isolated from term pregnant women as a model. Using a multiplex assay, HUSMCs treated with PGF2α changed their output of a number of cytokines and chemokines, with a distinct response pattern that differed between HUSMCs isolated from the upper and lower segment region of the uterus. Confirmatory enzyme-linked immunosorbent assays (ELISAs) showed that PGF2α stimulated increased output of interleukin (IL) 1β, IL6, IL8 (CXCL8) and monocyte chemotactic protein-1 (MCP1, also known as chemokine (c-c motif) ligand 2, CCL2) by HUSMCs isolated from both upper and lower uterine segments. In contrast, PGF2α inhibited tumor necrosis factor α (TNFα) release by HUMSCs from the lower uterine segment while the output of TNFα was undetectable in the upper segment. Small interfering (si) RNA mediated knockdown of the PGF2α receptor prevented the changes in cytokine and chemokine output by the HUSMCs. Since the PGF2α receptor (PTGFR) couples via the Gq protein and subsequently activates the phospholipase C (PLC) and protein kinase C (PKC) signaling pathways, we examined the role of these pathways in PGF2α modulation of the cytokines. Inhibition of PLC and PKC reversed the effects of PGF2α. PGF2α activated multiple signaling pathways including extracellular signal-regulated kinases (ERK) 1/2, phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K), P38, calcineurin/nuclear factor of activated T-cells (NFAT) and NF-κB signaling. Inhibition of ERK reversed PGF2α-induced IL1β, IL6 and CCL2 output, while inhibition of PI3K blocked the effect of PGF2α on IL6, CXCL8 and CCL2 output and inhibition of NF-κB reversed PGF2α-induced IL1β and CCL2 output. NFAT was involved in PGF2α modulation of CCL2 and TNFα output. In conclusion, our results support a role of PGF2α in creating an inflammatory environment during the late stage of human pregnancy.

Dynamic Changes in the Myometrium during the Third Stage of Labor, Evaluated Using Two-Dimensional Ultrasound, in Women with Normal and Abnormal Third Stage of Labor and in Women with Obstetric Complications

Objective: To investigate dynamic changes in myometrial thickness during the third stage of labor. Methods: Myometrial thickness was measured using ultrasound at one-minute time intervals during the third stage of labor in the mid-region of the upper and lower uterine segments in 151 patients including: women with a long third stage of labor (n = 30), postpartum hemorrhage (n = 4), preterm delivery (n = 7) and clinical chorioamnionitis (n = 4). Differences between myometrial thickness of the uterine segments and as a function of time were evaluated. Results: There was a significant linear increase in the mean myometrial thickness of the upper uterine segments, as well as a significant linear decrease in the mean myometrial thickness of the lower uterine segments until the expulsion of the placenta (p < 0.001). The ratio of the measurements of the upper to the lower uterine segments increased significantly as a function of time (p < 0.0001). In women with postpartum hemorrhage, preterm delivery, and clinical chorioamnionitis, an uncoordinated pattern among the uterine segments was observed. Conclusion: A well-coordinated activity between the upper and lower uterine segments is demonstrated in normal placental delivery. In some clinical conditions this pattern is not observed, increasing the time for placental delivery and the risk of postpartum hemorrhage.

Effect of carbetocin, oxytocin and prostaglandin E2 and F2α on intrauterine pressure in cows in dioestrus and oestrus.

To describe the physiological activity of the myometrium in oestrus and dioestrus and the induced activity after medication in cows with particular reference to segmental differences. Six cows were given the pharmaceuticals carbetocin, oxytocin and prostaglandin (PG) F2α (dinoprost) intramuscularly and PGE2 intravenously. The physiological myometrial activity was recorded for 15 minutes and the induced activity for 105 minutes by using a transcervically attached pressure probe containing six pressure microtransducers. Lower pressures were measured in dioestrus compared to oestrus before (dioestrus 3.2±8.88 mmHg, oestrus 12.4±13.23 mmHg, p<0.0001) and after the drug administration. Carbetocin provoked the longest lasting effect (60 minutes in dioestrus, 75 minutes in oestrus) followed by PGE2 (45 minutes in dioestrus, 60 minutes in oestrus), PGF2α (30 minutes each) and oxytocin (15 minutes in oestrus only). In contrast to the other drugs carbetocin did not cause any pressure decrease beneath the base level after the primary pressure increase in dioestrus. In dioestrus the pressure before drug administration was significantly higher in the cervix (3.6±19.40 mmHg) and the uterine body (7.1±36.10 mmHg) than in the uterine horn (1.1±7.21 mmHg). Conversely, in oestrus the pressure in the uterine horn (16.6±17.73 mmHg) was significantly higher than in the uterine body (6.2±16.59 mmHg) and the cervix (10.4±17.91 mmHg). Drug administration in dioestrus caused a cornual pressure increase and the pressure in the uterine body decreased, whereas in oestrus the pressure increased in all uterine segments. The physiological frequency of the pressure waves in dioestrus was 5.2±3.02 in 15 minutes compared to 7.5±2.89 in 15 minutes in oestrus. No traceable changes of the contraction frequency were found after medication. Carbetocin caused the most enduring increase in intrauterine pressure in dioestrus and oestrus and may therefore be indicated best for therapeutic use. The tested drugs had the same effects on the various uterine segments and no effect on the contraction frequency.

P24.06: Early appearance of pathologic uterine ring associated with persistent abnormal fetal posture

A pathologic ring of the uterus, rare in modern obstetrics, is a constrictive junction of the thin lower uterine segment and the thick upper segment. Most reports describe the pathologic rings associated with labour dystocia and diagnosed after active labour or at the time of Caesarean delivery. A case is presented involving a pathologic uterine ring suspected since late second trimester, identified using fetal ultrasonography and fetal Magnetic Resonance Imaging (MRI), and confirmed at Caesarean delivery. Serial fetal ultrasound examinations have shown that the lower uterine segment, also complicated with uterine synechiae, was filled with most of amniotic fluid and the right arm of the fetus and the upper uterine segment was crowded with the rest parts of the fetus. Until 30+2 weeks of gestational age, although limited, movements of the right arm across the pathologic uterine ring and the right wrist joint were noted. Fetal MRI confirmed shoulder presentation of the fetus with the corresponding arm prolapsed downward into the thinned lower uterine segment and showed a constriction at the junction of the upper and lower uterine segments. Caesarean delivery with classic incision was performed because of preterm rupture of membranes with uncontrolled preterm labour occurred at 32+5 weeks of gestational age. At the time of Caesarean delivery, a transverse thickened muscular band was noted. A female infant, weighing 1,840 g, was delivered with Apgar scores of 4 and 8 at 1 and 5 minutes. The infant's right arm looked slightly edematous, but moved freely without limitations of joints. Typically, the diagnosis of pathologic uterine rings, with the lack of identifiable signs, was unavoidably delayed after active labour. However, if a uterine constriction ring is suspected early in pregnancy and is not due to the process of labour and longstanding, fetal ultrasonography combined with fetal MRI might help decide the timing and mode of delivery, and the method of uterine incision, if Caesarean delivery is indicated.

The First Clinical Trial of Uterus Transplantation: Surgical Technique and Outcome.

Uterus transplantation may become a treatment for uterine factor infertility, which affects > 200,000 women only in Europe. The only option for these women, with no functional uterus, to gain genetic motherhood is by a surrogate mother. The latter procedure is not allowed in most countries due to ethical, religious or legal reasons. Our team has developed a procedure for uterus transplantation from live donor, through extensive research in several animal species. In May 2012 we were granted human IRB approval for a clinical study of 9 human transplantation cases with the uterus from live donors. The recipients and donors had been investigated for >12 months before the trial. This included IVF treatment. The donors were mothers (n=5), close relatives (n=3) or family friend and the ages were 35-62 years. Eight recipients were born with no uterus (MRKH syndrome) and one had been hysterectomized 7 years prior to the study because of cervical cancer. The ages of the recipients were between 27 and 38 years. The surgery of the donor involved isolation of the uterus with vascular pedicles including the uterine vessels and segments of the internal iliac vessels. The surgery of the recipient involved bilateral end-to-side anastomosis to the external iliacs, vagino-vagino anastomosis and uterine fixation to several pelvic ligaments. The surgical procedures will be discussed in detail. Durations of donor and recipient surgery ranged from 10 to 13 h and 4 to 6 h, respectively. The recipients received standard induction and initial triple maintenance immunosuppression.The 2nd uterus donor was diagnosed with a uterovaginal fistula on postoperative day 10. All other donor surgeries went without complications. No immediate perioperative complication occured among the recipients. However, one uterus had to be removed around 72 h after surgery due to bilateral uterine vessel thrombosis. Another uterus was removed 3.5 months after surgery due to persistent intrauterine infection and later development of an intrauterine abscess. Menstruation started after around 1-2 months in 7 recipients. In conclusion, the results demonstrate the surgical feasibility of live uterus transplantation. It remains to be determined what the pregnancy potential is in a transplanted uterus. Pregnancy results of our study population are expected in 2014.

Vascularization of broad ligament of uterus and its relationship with fetal and placental development in gilts

The objective of this study was to evaluate the influence of vascular architecture of broad ligament of the uterus on fetal and placental development in gilts. Fifteen gilts DB-90 (DanBred) were divided into three groups according to gestational age at slaughter (50, 80, and 106 days). After slaughter, fetuses and placentas were collected, weighed, and measured. The uterine arterial system was detached by latex repletion for quantification of the number and diameter of the terminal vessels in different regions of the uterine horns (apex, middle region, and base). Fetal and placental measurements were statistically analyzed and correlated with the number and diameter of arteries in each uterine segment. No correlation was observed (P > 0.10) between the number and diameter of arteries destined to the uterus with the number or weight of fetuses or placental weight in any gestational group. It was observed (P < 0.05) that more vessels destined to the medium region of the uterine horns, independent of the gestational age or uterus side. At the 80th day of gestation, fetuses located at the base of the uterus have (P < 0.05) smaller cephalic and thoracic perimeters. It was concluded that there were differences in vascularization of broad ligament that irrigates the different uterine segments, but this was not sufficient to influence the development of fetuses in gilts. The middle region of the uterine horns was the segment with a greater number of vessels, regardless of gestational age.

Uterine artery smooth muscle contractions are mediated by PKC and NO signaling mechanisms in diet‐induced obese pregnant rats (700.8)

Pre‐existing obesity increases the risk of maternal and fetal complications during gestation and is associated with systemic vascular dysfunction. We hypothesized that diet‐induced obesity would impair uterine artery contractile responses in pregnant rats. Female rats were fed for 3 weeks prior to pregnancy and throughout gestation either normal chow (LN‐Preg, n=8) or normal chow plus lard and liquid sucrose (OB‐Preg, n=8). Contractile responses to phenylephrine (PE, 10‐8 – 3x10‐5 M) were measured in isolated uterine arterial segments in late pregnancy, in the presence and absence of inhibitors of ERK1/2 (PD98059, 10‐5 M), Rho kinase (ROCK, Y27632: 10‐6 M), protein kinase C (PKC, BIM: 10‐6 M) and nitric oxide synthase (NOS, L‐NAME: 10‐4 M). OB‐Preg uterine arteries had reduced sensitivity to PE compared to LN‐Preg arteries. Inhibition of PKC reduced sensitivity to PE in OB‐Preg but not in LN‐Preg uterine arteries (‐logEC50, OB‐Preg, PE: 5.75 ± 0.087 vs. PE+BIM: 5.35 ± 0.128, p&lt;0.05; LN‐Preg, PE: 6.07 ± 0.086, PE+BIM: 5.99 ± 0.119, p&gt;0.05), whereas inhibition of ERK1/2 or ROCK reduced sensitivity to PE in both groups. NOS inhibition increased contractile responses to PE in OB‐Preg arteries to a greater extend as compared to LN‐Preg vessels and abolished the differences between groups (‐logEC50 in the presence of L‐NAME, OB‐Preg, 6.19 ± 0.103 vs. LN‐Preg, 6.19 ± 0.079, p&gt;0.05). Pre‐existing obesity reduces uterine artery contractile responses to α1 adrenergic stimulation in late pregnancy and this response is mediated by PKC and NO signaling mechanisms.Grant Funding Source: Supported by NIH, SWHR

Vascular Tissue Contractility Changes Following Late Gestational Exposure to Multi-Walled Carbon Nanotubes or their Dispersing Vehicle in Sprague Dawley Rats.

Multi-walled carbon nanotubes (MWCNTs) are increasingly used in industry and in nanomedicine raising safety concerns, especially during unique life-stages such as pregnancy. We hypothesized that MWCNT exposure during pregnancy will increase vascular tissue contractile responses by increasing Rho kinase signaling. Pregnant (17-19 gestational days) and non-pregnant Sprague Dawley rats were exposed to 100 μg/kg of MWCNTs by intratracheal instillation or intravenous administration. Vasoactive responses of uterine, mesenteric, aortic and umbilical vessels were studied 24 hours post-exposure by wire myography. The contractile responses of the vessel segments were different between the pregnant and non-pregnant rats, following MWCNT exposure. Maximum stress generation in the uterine artery segments from the pregnant rats following pulmonary MWCNT exposure was increased in response to angiotensin II by 4.9 mN/mm2 (+118%), as compared to the naïve response and by 2.6 mN/mm2 (+40.7%) as compared to the vehicle exposed group. Following MWCNT exposure, serotonin induced approximately 4 mN/mm2 increase in stress generation of the mesenteric artery from both pregnant and non-pregnant rats as compared to the vehicle response. A significant contribution of the dispersion medium was identified as inducing changes in the contractile properties following both pulmonary and intravenous exposure to MWCNTs. Wire myographic studies in the presence of a Rho kinase inhibitor and RhoA and Rho kinase mRNA/protein expression of rat aortic endothelial cells were unaltered following exposure to MWCNTs, suggesting absent/minimal contribution of Rho kinase to the enhanced contractile responses following MWCNT exposure. The reactivity of the umbilical vein was not changed; however, mean fetal weight gain was reduced with dispersion media and MWCNT exposure by both routes. These results suggest a susceptibility of the vasculature during gestation to MWCNT and their dispersion media-induced vasoconstriction, predisposing reduced fetal growth during pregnancy.

Characterization of electrical and mechanical activities of rabbit uterus associated with the presence of capacitated and non-capacitated spermatozoa

To investigate the effects capacitated spermatozoa may exert upon motility of the rabbit uterus, both contractility and electrical activity (frequency and intensity) were measured in 3 distinctive uterine segments of anaesthetized does: horn (UH), uterotubal junction (UTJ) and tube (UT) after 1) natural mating, 2) infusion of either seminal plasma or PBS, 3) infusion of either capacitated or non-capacitated spermatozoa. Basal values were: 17.1, 15.7, 16.4 g (contractility, P&gt;0.05); 3.5, 3.5, 3.4 Hz (frequency, P&gt;0.05); 0.49, 0.50, 0.57 μV (intensity, P&gt;0.05) for UH, UTJ, UT, respectively. Seminal plasma caused an increase (P&lt;0.05) in the UH contractility: 26.3 vs. 11.7 (natural mating) and 17.0 g (PBS); it also caused a decrease (P&lt;0.05) in electrical intensity at the UTJ: 0.24 vs. 0.67 (natural mating) and 0.58 μV (PBS). The presence of either capacitated or non-capacitated spermatozoa caused no changes in contractility and electrical frequency in any of the uterine segments. However, there was a change in electrical intensity at UTJ (0.37 vs. 0.57 μV for non-capacitated and capacitated spermatozoa, respectively; P&lt;0.05). There were also differences between segments by treatment: UTJ (0.37) vs. UT (0.59 μV) for non-capacitated; UH (0.46) vs. UT (0.71 μV) for capacitated spermatozoa (P&lt;0.05). In conclusion, use of this experimental model showed that uterine electrical activity was slightly modified by the presence of capacitated spermatozoa.

Gene expression of BDNF and its receptors, TrkB and p75 in the uterus and oviduct of pregnant and non-pregnant ewes

To compare genes expression of BDNF and its receptors, TrkB and p75 between pregnant and non-pregnant uterine and oviductal tissues of ewes, semi-quantitative RT-PCR was performed. BDNF, TrkB and p75 genes are normally expressed in the ovine uterus and oviduct. The relative amounts of BDNF mRNA were increased in the uterine segments of the pregnant samples with compared to non-pregnant samples. This increasing was only significant (P<0.05) in the inter-caruncle segment and in the cervix, which was not detectable in non-pregnant samples. In contrast, the relative amounts of BDNF mRNA were decreased in the oviductal segments of the pregnant samples and were undetectable in the isthmus. Variations of the relative amounts of TrkB and p75 mRNAs between the pregnant and non-pregnant samples were not significant (P>0.05) although TrkB mRNA in the caruncle and cervix and p75 mRNA in the cervix of pregnant samples were too less to be detected by RT-PCR method. It is probably that these changes of BDNF/receptors genes expression are necessary for normal pregnancy, and abnormal variations in the expression of these genes may be involved in the pathophysiological conditions.