Junctional Zone Research Articles

P-476 Metformin ameliorated testosterone-induced ferroptosis via inhibiting ferritinophagy in trophoblasts of polycystic ovary syndrome (PCOS) placentas

Abstract Study question Does trophoblasts of PCOS patients with aberrant iron metabolism, and how does metformin ameliorate androgen-induced ferroptosis in trophoblasts in vivo and in vitro? Summary answer Metformin ameliorates placental development and attenuates spontaneous miscarriage rate in early gestation by inhibiting ferritinophagy in trophoblasts of PCOS patients with hyperandrogenism. What is known already PCOS is a common reproductive endocrine disease in women of reproductive age, and a large proportion of patients present with hyperandrogenism. Studies have found that the incidences of spontaneous miscarriage and adverse pregnancy outcomes in PCOS patients are higher than those in healthy controls, which placental dysfunction is an important factor. It is related to mitochondrial dysfunction, oxidative stress and metabolic disorders. Ferroptosis is an iron-dependent and phospholipid peroxidation-mediated cell death. Studies have shown that iron ions participate in a variety of biochemical reactions in humans, and its abnormal metabolism may lead to some diseases especially in the reproductive system. Study design, size, duration Villi at gestational weeks of 6-8 were collected from 18 healthy controls and 18 women with PCOS. Human trophoblast stem cells (hTSCs) were isolated from fresh villi (6-8 weeks, n = 3) and cultured with testosterone, ferroptosis or autophagy inhibitors, metformin, or NCOA4 small interfering RNA. A dihydrotestosterone (DHT)-induced PCOS C57BL/6 mouse model was treated with or without metformin (3 months before mating) (n = 20 per group). Mouse placentas were obtained at embryonic days 11.5 (n = 50 per group). Participants/materials, setting, methods The levels of iron ions and lipid peroxidation were measured by PGSK staining, BODIPYC11 staining and MDA assay kit. The expressions of ferroptosis-related genes and proteins were assessed by qPCR, western blot and immunofluorescence. The binding of androgen receptor (AR) and NCOA4 to the putative androgen response element (ARE) was examined by ChIP, EMSA and luciferase assay. The influence of metformin on ferritinophagy in placentas of PCOS mice was confirmed by western blot and immunofluorescence. Main results and the role of chance The levels of iron ions and lipid peroxidation in villi at gestational weeks of 6-8 were higher in PCOS patients than in healthy controls. In vitro experiments, testosterone-induced ferroptosis was ameliorated by ferroptosis inhibitors, and ferritinophagy was mitigated by autophagy inhibitors or NCOA4 knockdown in primary cultured hTSCs. Testosterone induced ARs into the nucleus, where the recruitment of AR homodimers along with NCOA4 to the positive ARE on the SQSTM1 promoter, leading to autophagy induction and degradation of ferritin heavy chain 1 (FTH1) in autophagosomes, resulting in the release of ferritin bound irons as free ferrous irons. Metformin ameliorated testosterone-induced ferroptosis in hTSCs to a certain extent by inhibiting ferritinophagy. Compared to those of control mice, the placentas of DHT-treated PCOS mice showed increased levels of iron ions and lipid peroxidation, and altered expressions of ferroptosis-related proteins. Hyperandrogenism also induced ferritinophagy in labyrinthine zone of placentas in PCOS mice. Metformin ameliorated placental development was associated with decreased iron ions accumulation by inhibiting ferritinophagy in PCOS mouse placentas, which observably attenuated embryo absorption rate and spontaneous miscarriage incidence in the early gestation of PCOS mice. Limitations, reasons for caution The numbers of PCOS patients and mouse placentas in this study were small. The placental labyrinthine zone was not separated from the endocrine junctional zone and maternal decidua for molecular analyses. In addition, it would be better to isolated and cultured mouse TSCs to explore mechanisms on ferritinophagy in further. Wider implications of the findings This study provides further evidence for metformin as a PCOS treatment, which ameliorates placental development and attenuates spontaneous miscarriage rate in early gestation. Trial registration number not applicable

175-LB: Altered Glycogen Cell Number in Placental mTOR Signaling Deficiency Model in Mice

Fetal Growth Restriction (FGR) is a determinant for the development of chronic diseases, such as type II diabetes in adulthood. FGR is caused by the failure of the placenta to provide adequate nutrients and oxygen for fetal development. mTOR regulates cell metabolism and growth, and it is reduced in the FGR placenta. We previously reported that placental mTOR ablation induces insulin resistance and glucose intolerance in adult mice. We hypothesize that mTOR signaling regulates glucose utilization in the placenta, which stores glucose reservoirs as glycogen. Appropriate localization and breakdown of glycogen cells is crucial for normal fetal growth. We leveraged our mouse model lacking placental mTOR signaling to ask whether glycogen cell number was altered. First, we found that critical glycogenesis and glycogenolysis enzymes, as well as the glucagon receptor, were expressed in placental tissue, indicating the capability for glycogen cell synthesis and breakdown, like glycogen-rich tissues such as the liver. Next, we show increased glycogen cell counts in the junctional zone of the mTORKO placenta compared to littermate controls by IHC. However, whole placental glycogen content, measured by ELISA, demonstrates no changes between KO and control. These data suggest that mTOR signaling modulates the localization and migration of glycogen cells during high metabolic needs. Glucagon mRNA is expressed in the placenta, with significant increases in the male KO placenta, indicating the placenta may produce its own glucagon. However, IHC staining indicates increased glucagon levels in female KO placenta in the labyrinth zone. Together, these data suggest that the placenta may have the capability to regulate its own glycogen breakdown by altering glucagon expression. Our results indicate that the FGR placenta induced by mTOR deletion has altered glycogen cell content and mislocalization, which mimics the phenotypes of placenta impacted by gestational diabetes, suggesting a shared mechanistic pathway. Disclosure B. Clifton: None. M. Beetch: None. E. Alejandro: None. Funding National Institutes of Health (R01DK115720)

MRI for Diagnosing Intrauterine Adhesions

ObjectivesTo prospectively assess the diagnostic accuracy of MRI and transvaginal ultrasound (TVS) as well as the prognostic value of MRI for intrauterine adhesions (IUAs), using hysteroscopy as the reference standard. DesignProspective observational study. SettingTertiary medical centre. Patient(s)Ninety-two women with amenorrhea, hypomenorrhea, subfertility, or recurrent pregnancy loss who underwent MRI and in whom Asherman’s syndrome was suspected upon TVS. Intervention(s)MRI and TVS were conducted approximately 1 week before hysteroscopy. MethodsNinety-two patients suspected of having Asherman’s syndrome were examined by MRI and TVS within 7 days of an upcoming hysteroscopy. All hysteroscopy procedures were performed during the early proliferative phase of the menstrual cycle. All hysteroscopic diagnoses were performed by an experienced expert. All MRIs were read by 2 experienced, blinded radiologists. ResultsMRI was highly accurate (94.57%), sensitive (98.8%), and specific (42.9%) for diagnosing IUAs with a positive predictive value of 95.5% and a negative predictive value of 75%. The diagnostic values of MRI and TVS were significantly different according to McNemar tests. Junctional zone signal and junctional zone alterations correlated with the stage of IUAs. ConclusionMRI is markedly superior to TVS in terms of diagnostic accuracy for IUAs, with total agreement with hysteroscopic findings. However, the main advantage of MRI is that, unlike TVS and hysterosalpingography, it can be used to assess the risk of hysteroscopy and to predict postoperative recovery and future pregnancy based on the uterine junctional zone.

Uterine junctional zone and adenomyosis: comparison of MRI, transvaginal ultrasound and histology.

The uterine junctional zone is the subendometrial area in the myometrium that contributes to peristalsis and aids in spermatozoa and blastocyst transport. Alterations in the appearance of the junctional zone on transvaginal sonography (TVS) or magnetic resonance imaging (MRI) are associated with adenomyosis. The lack of standardization of description of its appearance and ill-defined boundaries on both histology and imaging hamper understanding of the junctional zone and limit its role in the diagnosis of adenomyosis. The objectives of this review were to investigate the accordance in definition of the junctional zone across different diagnostic approaches and to examine how imaging findings can be linked to histological findings in the context of diagnosis of adenomyosis. A comprehensive literature review was conducted of articles describing the appearance on imaging and the histological structure of the uterine junctional zone. Our review suggests that the junctional zone is distinguished from the middle and outer myometrium by gradual changes in smooth-muscle cell density, extracellular space, connective tissue, water content and vascular properties. However, while the signal intensity from the junctional zone to the middle myometrium changes abruptly on MRI, the histopathological changes are gradual and its border may be difficult or impossible to distinguish on two-dimensional TVS. Moreover, the thickness of the junctional zone measured on MRI is larger than that measured on TVS. Thus, these two imaging modalities reflect this zone differently. Although a thickened junctional zone is often used to diagnose adenomyosis on MRI, the presence of adenomyosis can be described more accurately as interruptions of the junctional zone by endometrial tissue, which leads to direct signs on imaging such as subendometrial lines and buds on two- and three-dimensional TVS or bright foci on MRI. The histopathological criteria for diagnosis are based on enlargement of the uterus with severe adenomyosis, and might not reflect its early stages. Clinicians should be aware that findings on MRI cannot be extrapolated readily to ultrasound. An understanding of this is necessary when investigating the uterine junctional zone as a functional unit and the association between visualization of direct features of adenomyosis in the junctional zone and clinical symptoms. © 2022 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Prediction of adenomyosis diagnosis based on MRI

ObjectiveDevelopment of a multivariate prediction model based on MRI and clinical parameters for histological adenomyosis diagnosis. Materials and methodsThis single centre retrospective cohort study took place in the gynaecological department of a referral hospital. In all, 296 women undergoing hysterectomy with preoperative pelvic MRI between 2007–2022 were included. MRI scans were retrospectively assessed for adenomyosis markers (junctional zone [JZ] parameters, high signal intensity [HSI] foci in a blinded fashion. A multivariate regression model for histopathological adenomyosis diagnosis was developed based on MRI and clinical variables from univariate analysis with p<0.1 and factors deemed clinically relevant. Results131/296 women (44.3%) had histopathological adenomyosis. Patients had comparable age at hysterectomy, BMI and clinical symptoms, p>0.05. Adenomyosis patients more often had: undergone a curettage (22.1% vs. 8.9%, p=0.002), a higher mean JZ thickness (9.40 vs. 8.35mm, p<.001), maximal JZ thickness (16.00 vs. 13.40mm, p<.001), mean JZ/myometrium ratio (0.56 vs. 0.49, p=.040), and JZ differential (8.60 vs. 8.15mm, p=.003). Presence of HSI foci was the strongest predictor for adenomyosis (39.7% vs. 8.9%, p<.001). Based on the parameters age and BMI, history of curettage, dysmenorrhoea, abnormal uterine bleeding (AUB), mean JZ, JZ differential≥5mm, JZ/myometrium ratio>40, and presence of HSI foci, a predictive model was created with a good area under the curve (AUC) of .776. ConclusionsThis is the first study to create a diagnostic tool based on MRI and clinical parameters for adenomyosis diagnosis. After sufficient external validation, this model could function as a useful clinical decision-making tool in women with suspected adenomyosis.

LncRNA Tuna is activated in cadmium-induced placental insufficiency and drives the NRF2-mediated oxidative stress response.

Cadmium (Cd) is a toxic heavy metal found throughout the environment and one of the top ten toxicants of major public health concern identified by the World Health Organization. In utero Cd exposure causes fetal growth restriction, malformation, and spontaneous abortion; however, the mechanisms by which Cd impacts these outcomes are poorly understood. Cd accumulates in the placenta, suggesting that these negative outcomes may be a consequence of disrupted placental function and placental insufficiency. To understand the impact of Cd on gene expression within the placenta, we developed a mouse model of Cd-induced fetal growth restriction through maternal consumption of CdCl2 and performed RNA-seq on control and CdCl2 exposed placentae. The top differentially expressed transcript was the Tcl1 Upstream Neuron-Associated (Tuna) long non-coding RNA, which was upregulated over 25-fold in CdCl2 exposed placentae. Tuna has been shown to be critical for neural stem cell differentiation. However, within the placenta, there is no evidence that Tuna is normally expressed or functional at any developmental stage. To determine the spatial expression of Cd-activated Tuna within the placenta, we used in situ hybridization as well as placental layer-specific RNA isolation and analysis. Both methods confirmed the absence of Tuna expression in control samples and determined that Cd-induced Tuna expression is specific to the junctional zone. Since many lncRNAs regulate gene expression, we hypothesized that Tuna forms part of the mechanism of Cd-induced transcriptomic changes. To test this, we over-expressed Tuna in cultured choriocarcinoma cells and compared gene expression profiles to those of control and CdCl2 exposed cells. We demonstrate significant overlap between genes activated by Tuna overexpression and genes activated by CdCl2 exposure, with enrichment in the NRF2-mediated oxidative stress response. Herein we analyze the NRF2 pathway and show that Tuna increases NRF2/NRF2 both at the transcript and protein levels. Tuna drives increased NRF2 target gene expression, a result that is abrogated with the use of an NRF2 inhibitor, confirming that Tuna activates oxidative stress response genes through this pathway. This work identifies the lncRNA Tuna as a potential novel player in Cd-induced placental insufficiency.

Expanded directly binds conserved regions of Fat to restrain growth via the Hippo pathway.

The Hippo pathway is a conserved and critical regulator of tissue growth. The FERM protein Expanded is a key signaling hub that promotes activation of the Hippo pathway, thereby inhibiting the transcriptional co-activator Yorkie. Previous work identified the polarity determinant Crumbs as a primary regulator of Expanded. Here, we show that the giant cadherin Fat also regulates Expanded directly and independently of Crumbs. We show that direct binding between Expanded and a highly conserved region of the Fat cytoplasmic domain recruits Expanded to the apicolateral junctional zone and stabilizes Expanded. In vivo deletion of Expanded binding regions in Fat causes loss of apical Expanded and promotes tissue overgrowth. Unexpectedly, we find Fat can bind its ligand Dachsous via interactions of their cytoplasmic domains, in addition to the known extracellular interactions. Importantly, Expanded is stabilized by Fat independently of Dachsous binding. These data provide new mechanistic insights into how Fat regulates Expanded, and how Hippo signaling is regulated during organ growth.

277 HPV8 E6 induced STAT3 activation leads to hair follicle junctional zone keratinocyte stem cell proliferation and expansion in actinic keratoses

277 HPV8 E6 induced STAT3 activation leads to hair follicle junctional zone keratinocyte stem cell proliferation and expansion in actinic keratoses

Intraoperative Computed Tomography Scanner-Guided Junctional Zone Spinal Surgery in Fluoroscopic Limitation Zone-An Early Experience

Objective: Confirmation of the level for surgery at the junctional zones of the spine is associated with unique difficulties. C-Arm has its limitations in these areas. The use of intraoperative CT scan with navigation greatly helps in the surgery at junctional zones. These junctional zones include the craniocervical junction, cervico-dorsal junction, and sacroiliac junction. In 2019, we installed an Airo Brain Lab 32-slice MDCT in our facility and researched to evaluate its advantages in junctional zone fluoroscopy-restricted spine operations. Material and Methods: We performed 20 complex surgeries in all these 3 junctional zones with the use of an intraoperative computed tomography scanner. 3 patients had craniovertebral junction fixation under intra-op CT while 6 had lateral mass fixation. 3 patients had cervicodorsal junction fixation and 4 patients had lumbosacral fixation. Results: 2 out of 3patients with cervicodorsal fixation had correction of screws trajectory after usage of intra-op CT and 1 patient with lumbosacral fixation. All patients were discharged home with neurology better or the same as compared to pre-op. status while 13 patients had a favorable outcome. Conclusion: Intraoperative computed tomography scanner is opening new horizons for complex spinal surgeries. Keywords: , ,, .

The effect of tauroursodeoxycholic Acid (TUDCA) treatment on placental endoplasmic reticulum (ER) stress in a rat model of advanced maternal age.

Advanced maternal age (≥35 years) is associated with an increased risk of pregnancy complications such as fetal growth restriction and preeclampsia. We previously demonstrated poor pregnancy outcomes (reduced fetal body weight), altered vascular function, and increased expression of endoplasmic reticulum (ER) stress markers (phospho-eIF2α and CHOP) in mesenteric arteries from a rat model of advanced maternal age. Further, treatment of aged dams during pregnancy with an ER stress inhibitor, tauroursodeoxycholic acid (TUDCA) increased fetal body weight (both male and female), tended to improve uterine artery function, and reduced expression of phospho-eIF2α and CHOP in systemic arteries. Placental ER stress has been linked to poor pregnancy outcomes in complicated pregnancies but whether placental ER stress is evident in advanced maternal age is not known. In addition, sex-specific changes in the placental labyrinth and junctional zones from male and female offspring in advanced maternal age have not been investigated. Therefore, the current study aimed to investigate the effect of TUDCA intervention on placental ER stress. We hypothesize that placental ER stress is increased in a rat model of advanced maternal age that is alleviated by TUDCA intervention for both sexes. Placental ER stress markers (GRP78, phospho-eIF2α, ATF-4, CHOP, ATF-6α, and sXBP-1) were quantified by Western blot in placentas from male and female offspring; the labyrinth and junction zones were analyzed separately. In the placental labyrinth zone from male offspring, only GRP78 (p = 0.007) was increased in aged dams compared to young dams; TUDCA treatment reduced the placental expression of GRP78 in aged dams (p = 0.003). In addition, TUDCA reduced the levels of phospho-eIF2α (p = 0.021), ATF-4 (p = 0.016), and CHOP (p = 0.012) in aged dams but no effect was observed in young TUDCA-treated dams. In the placental labyrinth zone from female offspring, an increased level of phospho-eIF2α (p = 0.005) was observed in aged dams compared to young dams, and TUDCA treatment had no effect in both young and aged groups. In the placental junctional zone from male and female offspring, no changes in the expression of GRP78, phospho-eIF2α, ATF-4, CHOP, and ATF-6α was observed with or without TUDCA treatment in both young and aged groups, however, a reduced expression of sXBP-1 protein was observed in from both male (p = 0.001) and female (p = 0.031) placentas from aged-TUDCA treated dams compared to aged control. In conclusion, our data highlight the complexity and sex-specificity of ER stress responses in advanced maternal age with TUDCA treatment maintaining ER stress proteins to basal levels and improving fetal growth in both male and female offspring.

Gse1, a component of the CoREST complex, is required for placenta development in the mouse

Gse1 is a component of the CoREST complex that acts as an H3K4 and H3K9 demethylase and regulates gene expression. Here, we examined the expression and role of Gse1 in mouse development. Gse1 is expressed in male and female germ cells and plays both maternal and zygotic roles. Thus, maternal deletion of Gse1 results in a high incidence of prenatal death, and zygotic deletion leads to embryonic lethality from embryonic day 12.5 (E12.5) and perinatal death. Gse1 is expressed in the junctional zone and the labyrinth of the developing placenta. Gse1 mutant (Gse1Δex3/Δex3) placenta begins to exhibit histological defects from E14.5, being deficient in MCT4+ syncytiotrophoblast II. The number of various cell types was largely maintained in the mutant placenta at E10.5, but several genes were upregulated in giant trophoblasts at E10.5. Placenta-specific deletion of Gse1 with Tat-Cre suggested that defects in Gse1Δex3/Δex3 embryos are due to placental function deficiency. These results suggest that Gse1 is required for placental development in mice, and in turn, is essential for embryonic development.

Low-dose letrozole – an effective option for women with symptomatic adenomyosis awaiting IVF: a pilot randomized controlled trial

Research questionCan low-dose letrozole reduce dysmenorrhoea, menorrhagia and sonographic features in symptomatic women with adenomyosis awaiting IVF? DesignThis was a longitudinal randomized prospective pilot study to explore the effectiveness of low-dose letrozole and compare it with a gonadotropin releasing hormone (GnRH) agonist in reducing dysmenorrhoea, menorrhagia and sonographic features in symptomatic women with adenomyosis awaiting IVF. The women were treated for 3 months, either with the GnRH agonist goserelin 3.6 mg/month (n = 77) or the aromatase inhibitor letrozole 2.5 mg three times weekly (n = 79). Dysmenorrhoea and menorrhagia were evaluated at randomization and followed up monthly using a visual analogue score (VAS) and pictorial blood loss assessment chart (PBAC), respectively. A quantitative scoring method was used to assess the improvement of sonographic features after 3 months of treatment. ResultsBoth groups reported a marked improvement in symptoms after 3 months of treatment. In both the letrozole and GnRH agonist groups, VAS and PBAC scores decreased significantly over the 3 months (letrozole: P = 0.0001 and P = 0.0001 for VAS and PBAC, respectively; GnRH agonist: P = 0.0001 and P = 0.0001 for VAS and PBAC, respectively). Participants on letrozole had regular menstruation cycles, while most of the women who received the GnRH agonist were amenorrhoeic, with only four women reporting mild bleeding. Haemoglobin concentrations also improved after both treatments (letrozole P = 0.0001, GnRH agonist P = 0.0001). A quantitative assessment of sonographic features showed significant improvements following both treatments (diffuse adenomyosis of the myometrium: letrozole P = 0.015, GnRH agonist P = 0.039; diffuse adenomyosis of the junctional zone: letrozole P = 0.025, GnRH agonist P = 0.001). Women with adenomyoma also responded well to both therapies (letrozole P = 0.049, GnRH agonist P = 0.024), whereas the letrozole group responded comparatively better in focal adenomyosis when the outer myometrium was involved (letrozole P < 0.001, GnRH agonist P = 0.26). No noticeable side effects were observed in women receiving letrozole therapy. Additionally, letrozole therapy was found to be more cost-effective than GnRH agonist treatment. ConclusionsLow-dose letrozole treatment is a low-cost alternative to a GnRH agonist, with comparable effects in improving the symptoms and sonographic features of adenomyosis in women awaiting IVF.

Effects of Stevioside on the Expressions of GLUT 1, GLUT 3, and GLUT 4 Proteins in Diabetic Rat Placenta.

The placenta provides maternal-fetal nutrient transport. The primary source of energy for fetus development is glucose and maternal-fetal glucose transport occurs through glucose transporters (GLUTs). Stevioside, a component of Stevia rebaudiana Bertoni, is used for medicinal and commercial purposes. We aim to determine the effects of stevioside on GLUT 1, GLUT 3, and GLUT 4 proteins expressions in diabetic rat placentas. The rats are divided into four groups. A single dose of streptozotocin (STZ) is administered to form the diabetic groups. Pregnant rats receive stevioside to form the stevioside and diabetic + stevioside groups. According to immunohistochemistry results, GLUT 1 protein is found in both the labyrinth and junctional zones. GLUT 3 protein is limited in the labyrinth zone. GLUT 4 protein is detected in trophoblast cells. According to Western blotting results, on the 15th and 20th days of pregnancy, there is no difference in the expression of GLUT 1 protein between groups. On the 20th day of pregnancy, the expression of GLUT 3 protein in the diabetic group is statistically higher compared to the control group. On the 15th day and 20th day of pregnancy, the expression of GLUT 4 protein in the diabetic group is statistically lower compared to the control group. Insulin levels in blood samples derived from rat abdominal aorta are determined by the ELISA method. According to the ELISA results, there is no difference in insulin protein concentration between groups. Stevioside treatment reduces GLUT 1 protein expression under diabetic conditions.

Maternal nano-titanium dioxide inhalation alters fetoplacental outcomes in a sexually dimorphic manner.

The placenta plays a critical role in nutrient-waste exchange between the maternal and fetal circulations, thus functioning as an interface that profoundly impacts fetal growth and development. The placenta has long been considered an asexual organ, but, due to its embryonic origin it shares the same sex as the fetus. Exposures to toxicant such as diesel exhaust, have been shown to result in sexually dimorphic outcomes like decreased placental mass in exposed females. Therefore, we hypothesize that maternal nano-TiO2 inhalation exposure during gestation alters placental hemodynamics in a sexually dimorphic manner. Pregnant Sprague-Dawley rats were exposed from gestational day 10-19 to nano-TiO2 aerosols (12.17 ± 1.69mg/m3) or filtered air (sham-control). Dams were euthanized on GD20, and fetal tissue was collected based on fetal sex: whole placentas, placental junctional zone (JZ), and placental labyrinth zone (LZ). Fetal mass, placental mass, and placental zone percent areas were assessed for sex-based differences. Exposed fetal females were significantly smaller compared to their exposed male counterparts (2.65 ± 0.03g vs 2.78 ± 0.04g). Nano-TiO2 exposed fetal females had a significantly decreased percent junctional zone area compared to the sham-control females (24.37 ± 1.30% vs 30.39 ± 1.54%). The percent labyrinth zone area was significantly increased for nano-TiO2 females compared to sham-control females (75.63 ± 1.30% vs 69.61 ± 1.54%). Placental flow and hemodynamics were assessed with a variety of vasoactive substances. It was found that nano-TiO2 exposed fetal females only had a significant decrease in outflow pressure in the presence of the thromboxane (TXA2) mimetic, U46619, compared to sham-control fetal females (3.97 ± 1.30mm Hg vs 9.10 ± 1.07mm Hg) and nano-TiO2 fetal males (9.96 ± 0.66mm Hg). Maternal nano-TiO2 inhalation exposure has a greater effect on fetal female mass, placental zone mass and area, and adversely impacts placental vasoreactivity. This may influence the female growth and development later in life, future studies need to further study the impact of maternal nano-TiO2 inhalation exposure on zone specific mechanisms.

Adenomyosis: In-Vitro Fertilization Does Not Improve Pregnancy Outcome as Compared to Conventional Infertility Treatment

Adenomyosis is a benign invasion of endometrial glands and stroma into the uterine myometrium. [1, 2]. It can present with heavy and painful menstrual bleeding or be asymptomatic [2-4]. Over last three decades the diagnosis of adenomyosis has been feasible after the introduction of MRI and 3D-TVS. Although there are no international consensus on the diagnostic criteria for adenomyosis with either ultrasound or MRI, the Morphological Uterus Sonographic Assessment (MUSA) consensus statement provides several important ultrasounds features for diagnosis of adenomyosis [5-8]. Heterogeneous and hypoechogenic areas in the myometrium, areas with or without anechoic lacunae or cysts of varying size, linear striation radiating out from the endometrium into the myometrium, poor definition of the junctional zone (JZ), and pseudo-widening of the endometrium (enlargement of uterus with asymmetric thickening of the anterior or posterior walls).

Lectin histochemistry reveals two cytotrophoblast differentiation pathways during placental development in the feline (Feliscatus)

IntroductionPlacental glycosylation has been examined on eight feline placentae ranging from approximately 15 to 60 days post-conception as little is known about changes in glycan distribution in this species. MethodsSpecimens were resin embedded and lectin histochemistry was applied to semi-thin sections using a panel of 24 lectins and an avidin-biotin revealing system. ResultsAbundant tri-tetraantennary complex N-glycan and α-galactosyl residues found in the syncytium in early pregnancy were greatly reduced in mid-pregnancy, though retained at the invasion front in the syncytium (N-glycan) or cytotrophoblast layer (αGal). Some other glycans were also uniquely present in invading cells. Abundant polylactosamine was found in the infolding basal lamina of syncytiotrophoblast and the apical villous cytotrophoblast membrane. Syncytial secretory granules often clustered near the apical membrane abutting maternal vessels. Decidual cells selectively expressed β-galactosyl residues throughout pregnancy and highly branched N-glycan levels increased over time. DiscussionGlycan distribution changes significantly over pregnancy, probably relating to the development of transport and invasive properties of trophoblast which in the endotheliochorial placenta reaches the level of the maternal vessels. Highly branched complex N-glycans, often associated with invasive cells, N-Acetylgalactosamine and terminal α-galactosyl residues are present at the invasion front abutting the junctional zone of the endometrium. Abundant polylactosamine on the syncytiotrophoblast basal lamina may reflect the presence of specialised adhesive interactions, while clustering of glycosylated granules apically is probably associated with secretion and absorption of material via maternal vasculature. It is suggested that lamellar and invasive cytotrophoblast represent distinct differentiation pathways. 246 words.

A rapid wide complex tachycardia

A rapid wide complex tachycardia

Diagnostic Accuracy of GATA6 Immunostaining in Sebaceous Tumors of the Skin

The accurate diagnosis of skin adnexal neoplasms is sometimes challenging but is necessary because medical management and follow-up may differ between tumors. GATA6 transcription factor has been identified as a new marker of the upper folliculosebaceous compartment (lower infundibulum, junctional zone and isthmus, and upper sebaceous gland) in the human skin. We aimed to determine the diagnostic accuracy of GATA6 immunostaining to diagnose sebaceous tumors compared with that to diagnose other adnexal and nonadnexal cutaneous neoplasms. We conducted a retrospective, evaluator-nonblinded study comparing the reference standard (diagnosis by an expert dermatopathologist) with GATA6 immunostaining to identify sebaceous tumors in a cohort containing 234 different tumors. The GATA6 expression score was significatively higher in sebaceous than that in nonsebaceous tumors. In addition, tumors originating from the upper hair follicle showed positive results for GATA6 staining; however, they showed lower GATA6 expression scores. Detection of sebaceous tumors using GATA6 positivity had a sensitivity of 95.7% (95% CI, 85.8-99.2), specificity of 80.8% (95% CI, 74.5-85.8), positive predictive value of 55.6% (95% CI, 44.7-65.9), and negative predictive value of 98.7% (95% CI, 95.4-99.8). GATA6 showed similar sensitivity to adipophilin, the reference marker; however, the specificity of GATA6 was higher, as observed in a cohort of 106 tumors enriched in squamous cell carcinomas with clear-cell histology. In addition, GATA6 positivity was assessed in 39 sebaceous carcinomas and compared with epithelial membrane antigen (EMA), CK7, and androgen receptor (AR) staining results. Although CK7 staining displayed lower diagnostic performances, GATA6 staining showed comparable results as EMA and AR. Finally, we found GATA6 expression in skin metastases of gastrointestinal origin, whereas GATA6 was absent in metastases originating from breast or lung cancers. Overall, our work identified GATA6 immunostaining as a new diagnostic tool for sebaceous tumors.

The trophoblast giant cells of cricetid rodents.

Giant cells are a prominent feature of placentation in cricetid rodents. Once thought to be maternal in origin, they are now known to be trophoblast giant cells (TGCs). The large size of cricetid TGCs and their nuclei reflects a high degree of polyploidy. While some TGCs are found at fixed locations, others migrate throughout the placenta and deep into the uterus where they sometimes survive postpartum. Herein, we review the distribution of TGCs in the placenta of cricetids, including our own data from the New World subfamily Sigmodontinae, and attempt a comparison between the TGCs of cricetid and murid rodents. In both families, parietal TGCs are found in the parietal yolk sac and as a layer between the junctional zone and decidua. In cricetids alone, large numbers of TGCs, likely from the same lineage, accumulate at the edge of the placental disk. Common to murids and cricetids is a haemotrichorial placental barrier where the maternal-facing layer consists of cytotrophoblasts characterized as sinusoidal TGCs. The maternal channels of the labyrinth are supplied by trophoblast-lined canals. Whereas in the mouse these are lined largely by canal TGCs, in cricetids canal TGCs are interspersed with syncytiotrophoblast. Transformation of the uterine spiral arteries occurs in both murids and cricetids and spiral artery TGCs line segments of the arteries that have lost their endothelium and smooth muscle. Since polyploidization of TGCs can amplify selective genomic regions required for specific functions, we argue that the TGCs of cricetids deserve further study and suggest avenues for future research.

The AKT1-FOXO4 axis reciprocally regulates hemochorial placentation.

Hemochorial placentation involves the differentiation of invasive trophoblast cells, specialized cells that possess the capacity to exit the placenta and invade into the uterus where they restructure the vasculature. Invasive trophoblast cells arise from a well-defined compartment within the placenta, referred to as the junctional zone in rat and the extravillous trophoblast cell column in human. In this study, we investigated roles for AKT1, a serine/threonine kinase, in placental development using a genome-edited/loss-of-function rat model. Disruption of AKT1 resulted in placental, fetal and postnatal growth restriction. Forkhead box O4 (Foxo4), which encodes a transcription factor and known AKT substrate, was abundantly expressed in the junctional zone and in invasive trophoblast cells of the rat placentation site. Foxo4 gene disruption using genome editing resulted in placentomegaly, including an enlarged junctional zone. AKT1 and FOXO4 regulate the expression of many of the same transcripts expressed by trophoblast cells, but in opposite directions. In summary, we have identified AKT1 and FOXO4 as part of a regulatory network that reciprocally controls critical indices of hemochorial placenta development.