bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Major Impact of Obesity on the Development of Type 2 Diabetes (T2D) in Women With PCOS: A Systematic Review and Meta-Analysis of Observational Studies
Authors: Anagnostis Panagiotis , Paparodis Rodis , Bosdou Julia , Bothou Christina , Goulis Dimitrios G , Macut Djuro P , Dunaif Andrea Elizabeth , Livadas Sarantis 
Published: May 2021

Abstract
Background/Aims: Polycystic ovary syndrome (PCOS) is associated with disordered carbohydrate metabolism and an increased risk for T2D. However, there are limited data on the magnitude of this risk. Furthermore, 50-80% of women with PCOS are obese and obesity is known to have a synergistic deleterious effect on glucose tolerance in affected women. We systematically reviewed the literature regarding the association between PCOS, obesity and T2D risk. Methods: A comprehensive search was conducted in PubMed, CENTRAL and Scopus databases. Data are expressed as relative risk (RR) with 95% confidence intervals (CI). The I2 index was employed for heterogeneity. The available data, did not allow us to analyze the impact of weight status as normal, overweight and obese and as a consequence the studied subjects were stratified as obese (BMI&gt;30 kg/m2) and non-obese (BMI&lt;30kg/m2). Results: Twelve studies fulfilled eligibility criteria, yielding a total of 224,284 participants (45,361 PCOS and 5,717 T2DM cases). Women with PCOS had a higher risk of T2D compared with to unaffected women (RR 3.13, 95% CI, 2.83-3.47, p&lt;0.001; I2 40.1%). When women with PCOS were stratified according to the presence or absence of obesity, the RR for developing T2D in obese compared with non-obese women with PCOS was 4.20 (95% CI 1.97-9.10; p&lt;0.001). Moreover, compared to control women, the RR for developing T2D was significantly increased only in obese PCOS, RR 4.06 (95% CI 2.75-5.98; p&lt;0.001). There was a trend toward significantly increased risk in non-obese PCOS women [RR 2.68 (95% CI 0.97-7.49; p=0.06). Conclusion: Women with PCOS have a &gt;3-fold increased risk of T2D compared to women without PCOS, but this risk is substantially increased by the presence of obesity. Accordingly, weight reduction should be pursued in these women. References: 1. Dunaif A, Segal KR, Futterweit W, Dobrjansky A. Profound peripheral insulin resistance, independent of obesity, in polycystic ovary syndrome. Diabetes. 1989;38(9):1165-1174.2. Legro RS, Kunselman AR, Dodson WC, Dunaif A. Prevalence and predictors of risk for type 2 diabetes mellitus and impaired glucose tolerance in polycystic ovary syndrome: a prospective, controlled study in 254 affected women. J Clin Endocrinol Metab. 1999;84(1):165-169.3. Ehrmann DA, Barnes RB, Rosenfield RL, Cavaghan MK, Imperial J. Prevalence of impaired glucose tolerance and diabetes in women with polycystic ovary syndrome. Diabetes Care. 1999;22(1):141-146.4. Rubin KH, Glintborg D, Nybo M, Abrahamsen B, Andersen M. Development and risk factors of type 2 diabetes in a nationwide population of women with polycystic ovary syndrome. J Clin Endocrinol Metab. 2017;102(10):3848-3857.
Differential Expression Changes in Human Decidua With Term and Preterm Labor: Role for Upstream Targets in the Prostaglandin Pathway
Authors: Hornaday Kylie K , Wood Stephen L , Anderson David , Slater Donna M 
Published: May 2021

Abstract
Introduction: Prostaglandins (PGs) are paracrine mediators thought to be important during pregnancy and labor. Synthesis of PGs is complex with three rate limiting steps, the second of which (PTGS2/cyclooxygenase) is a target for managing preterm labor, but this treatment is not always effective. It is suggested that alternate steps in the PG pathway may play distinct roles during labor and hence the potential as alternate targets for labor management. The first step of PG synthesis involves the action of phospholipase A2 (PLA2) enzymes, of which over a dozen isotypes are known. However, PLA2 expression and role in the decidua (maternal-fetal interface) is unknown. We hypothesize that distinct PLA2 isotypes are present within the decidua and play a role in term and preterm labor. To this end, we conducted an expression profile of PLA2 subtypes and related PG genes within human decidua and determined differential expression patterns with labor at term and preterm. Methods: Decidual samples from term (40wks) and preterm (26-37wks) Cesarean deliveries, were used to assess PG gene expression by RNAseq and qRT-PCR (n=7 preterm non-labor, n=11 preterm labor, n=31 term non-labor, n=31 term labor). Analyses were conducted using student’s t-test and one-way ANOVA to compare labor (non-labor vs labor) or with gestation (preterm vs term) and adjusted for multiple testing using the Benjamini-Hochberg method. Results: RNAseq analysis identified 30 highly expressed PG genes in decidua, of which 12 had not been previously reported in the uterus, including 7 PLA2 isotypes. Confirming previous work, expression of PTGS2 was higher with labor (p=0.011). In contrast, we demonstrate that PLA2 isotype expression is lower with labor, with further differences between term and preterm samples. With term labor, expression of PLA2 isotype 2D was lower compared to non-labour (p=0.047). Meanwhile, preterm labor was associated with lower expression of PLA2 isotype 16 (p=0.025) and 4C (p=0.026), compared to preterm non-labour. Epression of PLA2 subtypes 2A, 15 and 4A late was highest in late labor (p=0.016, 0.004, 0.03, respectively). Conclusion: The presence of multiple PLA2 subtypes within the decidua suggests the potential for fine tuning PG synthesis during pregnancy. Expression of PLA2 isotypes 16 and 4C was uniquely associated with preterm labor, suggesting these isotypes may play a role in the pathogenesis of preterm labor. Further investigation of the functional role of PLA2 isotypes may provide insight to a novel mechanism for preterm labor and identify potential targets for labor management.
Expression of Cannabinoid Receptors in Mouse and Human Intrauterine Tissues: Insights into the Role of Endocannabinoid Signaling in Late Pregnancy
Authors: Urrego Daniela , Hornaday Kylie Karmila , Wood Stephen L , Slater Donna Michelle 
Published: May 2021

Abstract
Introduction: Endocannabinoid signaling (ECS), mediated primarily by cannabinoid receptors (CNR) 1 and 2, is implicated in embryo implantation, decidualization, and placentation, but less is known about its role in late pregnancy or labour. Reports of elevated serum endocannabinoid concentrations during labour suggest that ECS may modulate uterine function leading up to and during parturition. Effects on uterine function likely vary depending on: the type of cannabinoid present, the intrauterine tissue, and CNR1/2 expression. Study of ECS gene expression in late pregnancy is therefore important to determine the contribution of this pathway to the normal and pathologic physiology of labour, for instance in preterm labour. Examining ECS gene expression in pregnancy may also contribute insight into the effects of exogenous cannabinoid consumption (reported in ~5% of pregnancies) on the late-pregnant uterus. To understand when and where cannabinoid signaling may impact uterine functions, we conducted an observational study on pregnant human and mouse intrauterine tissues.
Methods: Human amnion, chorion, decidua, placenta, and upper/lower myometrium (n=6 participants) were biopsied at cesarean delivery (term, non-labour) to determine CNR expression by qPCR. Additional decidua (n=80) and upper/lower myometrium (n=82) samples were similarly obtained with and without labour, at term and preterm, for CNR expression analysis by qPCR. Mouse uteri for CNR expression analysis by qPCR were obtained from timed-mated C57BL6 mice at days 15-20 of pregnancy, in active labour, or post-partum. Human and mouse expression data were analyzed by Student’s t-test, one-way ANOVA (Bonferroni post-hoc), and Pearson’s correlation, as appropriate.
Results: Term non-labour chorion, placenta, myometrium and decidua express CNRs, with 2-fold higher expression of CNR1 versus CNR2. In myometrium and decidua, neither CNR1 or CNR2 expression differed with labour status. CNR1 expression correlated positively with gestational age in decidua (r=0.346, p=0.0266) and lower segment myometrium (r=0.3667, p=0.0270) with labour. In mouse uteri, Cnr1 and Cnr2 expression significantly increased post-partum compared to mid/late pregnancy (vs day 19, p&lt;0.0001 Cnr1; vs days 15-18, p&lt;0.05 Cnr2).
Conclusion: ECS may occur in the human decidua and myometrium throughout pregnancy and labour as CNR1 and CNR2 expression is maintained throughout. In our mouse model, higher Cnr1 and Cnr2 expression post-partum suggests that ECS may play a role in uterine resolution following delivery. Additional RNAseq analysis of the mouse tissues studied is underway to address whether expression of genes involved in production, transport or metabolism of cannabinoids are altered in late pregnancy or with the initiation of parturition.
Homeodomain Transcription Factors: Integral Modulators of Human Decidualization
Authors: Haller Meade , Yin Yan , Ma Liang 
Published: May 2021

Abstract
Ineffective embryo implantation accounts for a significant percentage of female infertility, and often renders IVF procedures unsuccessful. Decidualization, the dramatic uterine morphological response to ovarian hormone exposure, is a prerequisite for embryo implantation. Despite its significance in reproduction, the genetic framework of decidualization was not systematically studied until our recent development of a suitable high-throughput screening tool, immortalized human endometrial stromal cells (hESCs) that carry the yellow fluorescent protein gene under the control of the progesterone-sensitive prolactin promoter (PRL-Y cells). We recently used PRL-Y cells to perform a genome-wide siRNA functional screen and results revealed that 36 members of the homeodomain-containing family of transcription factors (HDTFs) are modulators of human decidualization. To determine which HDTFs are transcriptionally sensitive to ovarian hormone exposure, RT-PCR was performed on wildtype hESCs for the 36 HDTF hits over a 72-hour time course of E2/P4/cAMP exposure. Twenty HDTF hits (55%) were both detectable by PCR and showed variable expression in response to ovarian hormone treatment. Interestingly, all of these homeodomain factors, with a few distinct exceptions, exhibited decreased transcriptional expression in response to ovarian hormone treatment. This suggests that precious energy is used to transcribe these factors during the pre-decidualized phase, and that they may be required to maintain homeostasis during times of low hormone exposure. Because siRNA is not fully efficient, in order to confirm which HDTFs are required for normal decidualization, we generated a doxycyline-inducible Cas9-expressing hESC clone in order to subsequently generate individual knockout hESC lines for each HDTF hit. Cas9 expression was turned on 5 days prior to crRNA and trRNA transfection targeting the first exon of each HDTF. Five days after transfection, the cells were treated for 72 hours with ovarian hormone induction medium before RNA was isolated for gene expression analysis. As a pool of cells prior to any cloning (which likely includes knockout and wildtype cells in different ratios) the results indicate that several HDTFs are required for proper decidualization. The reporter transcripts of PRL and EREG are significantly abrogated or entirely undetectable in certain knockout lines. Interestingly these include some original siRNA HDTF hits whose expression is undetectably low by PCR. Sequencing validation will be necessary to confirm that knocking out such low levels of these transcripts genuinely has the robust effect on the human decidualization reaction that we are witnessing in these results. Together these findings comprise significant initial steps in characterizing the intricate upstream roles of HDTFs in human decidualization and female fertility.
Identification of Cellular Damage in Uteri From Hyperinsulinemic Mice Treated With Unopposed Estradiol
Authors: Kelly Katherine C , Huang Jane , Joshi Aditi A , Tal Aya , Wong Serena , Flannery Clare A 
Published: May 2021

Abstract
Unopposed estradiol and obesity are known risk factors for endometrial adenocarcinoma (EC). Endometrium from women with obesity was found to have an increase in mutations relative to tissue from normal weight women, indicating DNA damage may be accelerated in the setting of obesity. Since obesity is associated with high levels of insulin and anovulatory cycles, we sought to mimic these conditions in a mouse model. We previously found that hyperinsulinemic MKR mice, without the confounder of obesity, have an increased incidence of nuclear atypia in endometrial glands. We hypothesized that hyperinsulinemia and unopposed estradiol have a synergistic effect on inducing abnormal architecture and DNA damage in the endometrium, than either alone.
At 8-10 weeks old, cohorts of MKR (n=20) and WT (n=20) mice underwent ovariectomy and placement of either an estradiol (E2) or placebo (P) pellet. Metabolic profiling included insulin tolerance testing and MR for body composition. At 3 months post-implantation, mice received a partial hysterectomy and second pellet replacement. At 6 months, the remaining uterus was bisected into pieces. A blinded histological analysis was conducted by a gynecology pathologist. A marker of DNA damage due to oxidative stress, 8-oxoguanine-DNA-glycosylase (8-OHdG), was quantified by ELISA. Data was analyzed using Kruskal-Wallis test with multiple test correction, or Fischer’s exact test.
By 6 months, MKR-E2 treated mice had a 27% lower body weight than MKR-P mice (p&lt;0.05), and 31% lower than WT-E2 mice (p&lt;0.01). WT-E2 and WT-P had similar weight, and were similar to MKR-P (p=ns). Percent body fat was similar across all 4 cohorts of mice (p=ns). Since placebo-treated mice had small, atrophied uteri with minimal gland formation, E2 pellet failure was determined by the presence of small, atrophied uteri and occurred in 4 MKR and 3 WT mice at either 3 or 6 months. All other MKR and WT E2 treated mice had enlarged uteri. The frequency of endometrial gland dilation was similar in MKR-E2 and WT-E2 uteri (p=ns), but all MKR mice had moderate-severe dilation, whereas WT mice had 50% mild and 50% moderate-severe dilation (p=0.07). Focal hyperplasia was present in one MKR-E2 mouse, and nuclear atypia was present in one WT-E2 mouse. MKR-P uteri had a 7-fold higher mean 8-OHdG relative to MKR-E2 uteri (5.0±3.7 vs 0.7±1.6, p&lt;0.002). WT-E2 and WT-P uteri had similar 8-OHdG (1.6±0.8 vs 1.8±0.6, p=ns), as did MKR-E2 and WT-E2 uteri (p=ns).
Our findings show that hyperinsulinemia exacerbates the cystic dilation induced by chronic unopposed estradiol, indicating a synergy of insulin and estradiol in promoting abnormal glandular growth in the endometrium. Surprisingly, uterine DNA damage was highest in the setting of hyperinsulinemia alone, in a hormonal state mimicking post-menopause. Further work is needed to understand the effect of estradiol on intrauterine oxidative stress-induced damage.
Long-Term Effects of Late Gestation in Utero Hypoxic Stress on Mood Disorders: Sex and Age Differences
Authors: Mabry Steve , Wilson E Nicole , Rybalchenko Nataliya , Engelland Rachel , Fadeyibi Oluwadarasimi , Osikoya Oluwatobiloba , Cushen Spencer , Goulopoulou Styliani , Cunningham Rebecca Lynn 
Published: May 2021

Abstract
Introduction: In utero insults have been linked with increased fear and anxiety in progeny. In utero hypoxic stress is associated with a multitude of gestational complications such as pregnancy-associated hypertensive disorders and intrauterine growth restriction. Maternal hypertension during pregnancy is also associated with increased mood and anxiety disorders in progeny. However, it is unknown if these associations are due to in utero hypoxic stress. We hypothesized that exposure to late gestational hypoxia will have a long-term impact on anxiety in progeny. Methods: Timed pregnant female Long-Evans rats were exposed to five days (gestational days: 15-20) of chronic intermittent hypoxia (CIH) or room air (normoxia - 21% O2) for 8 hours during their sleep phase. Each CIH cycle was 6 min of 3 min hypoxia (10% O2) and 3 min normoxia (21% O2) for a total of 10 CIH cycles/hour. At weaning (PND 28), progeny was pair-housed with a conspecific of same sex and similar weight. To examine mood and anxiety disorders, we quantified anxiety-related behaviors (time spent in the center of open field arena, marble burying test, social and anti-social behaviors with conspecifics) along with quantifying food intake and circulating sex hormone levels during puberty (postnatal day, PND 40-45) and young adulthood (PND 60-65) in male and female progeny. Results: Gestational CIH did not impact circulating sex hormones or food intake, regardless of sex or age of progeny. However, gestational CIH increased anxiety related behaviors in pubertal females. These effects of gestational CIH on anxiety in pubertal females were not maintained, as these behaviors resolved in young adulthood. Gestational CIH did not impact male progeny, regardless of age. Conclusion: Exposure to CIH during gestation resulted in increased anxiety related behaviors in pubertal female progeny. In utero hypoxia during late gestation may temporarily increase the risk for mood and anxiety disorders in pubertal females.
Long-Term Effects of Late Gestation in Utero Hypoxic Stress on Neurodegeneration: Sex and Age Differences
Authors: Wilson E Nicole , Mabry Steve , Rybalchenko Nataliya , Engelland Rachel , Fadeyibi Oluwadarasimi , Osikoya Oluwatobiloba , Cushen Spencer , Goulopoulou Styliani , Cunningham Rebecca Lynn 
Published: May 2021

Abstract
Introduction: In utero insults have been proposed to lead to the onset of neurodegenerative diseases later in life, such as Parkinson’s disease (PD). In utero hypoxia is associated with a multitude of conditions, such as maternal sleep apnea, preeclampsia, gestational diabetes, and maternal hypertension. Exposure to in utero hypoxia may impact male progeny more than female progeny, which may underlie the male biased sex differences in PD. It is currently unknown whether late gestational hypoxic stress has a long-term effect on brain regions associated with PD, such as the nigrostriatal pathway. We hypothesized that exposure to late gestational hypoxia will result in nigrostriatal impairment in adult male progeny compared to adult female progeny. Methods: Timed pregnant female Long-Evans rats were exposed to five days (gestational days: 15-20) of chronic intermittent hypoxia (CIH) or room air (normoxia - 21% O2) for 8 hours during their sleep phase. Each CIH cycle was 6 min of 3 min hypoxia (10% O2) and 3 min normoxia (21% O2) for a total of 10 CIH cycles/hour. Gestational age at delivery was recorded and neonate’s body weights were measured within 12-16 hours from birth. At weaning (postnatal day, PND 28), progeny was pair-housed with a conspecific of the same sex and similar weight. To examine PD, we focused on PD associated characteristics of oxidative stress in the nigrostriatal pathway and behavioral impairments of motor (open field activity and ultrasonic vocalizations) and cognitive (spatial memory) function during puberty (PND 40-45) and young adulthood (PND 60-65). Results: Gestational CIH had no effect on the duration of gestation, litter size, and neonatal weight at birth. Gestational CIH did not impact circulating oxidative stress, regardless of sex or age of progeny. Offspring gross motor function (open field activity) and cognitive (Morris Water maze) function were unaffected by gestational CIH. In contrast, gestational CIH impaired ultrasonic vocalizations in adult male progeny. Gestational CIH increased the latency to vocalize and decreased the loudness of the vocalizations in adult male progeny. Conclusion: Exposure to CIH during gestation resulted in nigrostriatal impairment in adult male progeny, as evidenced by impaired ultrasonic vocalizations that require a functional nigrostriatal pathway. In utero hypoxia during late gestation may increase the risk for PD in males.
Adrenocortical Carcinoma Treatment in the Netherlands: An Analysis From the Netherlands Cancer Registry From 2014 to 2019
Authors: Steenaard Rebecca , Rutjens Marieke , Haak Harm Reinout 
Published: May 2021

Abstract
Background: Adrenocortical carcinoma (ACC) is a rare disease with often poor prognosis. Previous research has shown that surgery in a Dutch Adrenal Network (DAN) center increases the chance of survival. We aim to explore the determinants and survival of patients with ACC recently treated in the Netherlands both within and outside DAN centers. Methods: We analyzed retrospectively collected data from 172 adult patients with newly diagnosed ACC and 97 patients with recurrence or new metastases, registered between 2014 and 2019 in the Netherlands Cancer Registry. Differences in survival were analyzed with cox-regression analysis. Results: More than half of the new cases presented with advance disease (25.7% stage III, 34.6% stage IV) and the median survival was 29 months. The majority of treatments occurred within a DAN center (87.2% of surgery, compared to 56.4% between 1999 and 2009; and 94.5% of medical treatment). There were no differences in patient characteristics between the centers apart from a relatively high number of patients with stage IV disease outside DAN centers (47.2% vs. 28.7%). Adrenal resection and mitotane therapy both resulted in a significant survival benefit (resection HR 0.29, CI95%[0.17–0.49]; mitotane HR 0.61, CI95%[0.37–0.99], corrected for stage). Still, a remarkable proportion of patients with advanced disease received no mitotane treatment (39.8%). Due to the small number of patients treated outside DAN centers, survival benefits could not be tested. Conclusions: Centralization of ACC care in the Netherlands has improved since the previous report, but a further improvement in centralization of surgery can be made. Adrenal resection and mitotane treatment remain the main form of treatment, with a clear survival benefit. Further research is necessary to determine why mitotane treatment is withheld in a large proportion of patients with advance disease.