Are Twin Pregnancies at Higher Risk for Iron and Calcium Deficiency than Singleton Pregnancies?

The aim of this study was to compare the iron and calcium status in singleton and twin pregnancies and to assess whether there is an increased risk for iron and calcium deficiency in twin gestation. The study included 105 singleton and 9 twin pregnancies at or above 35 weeks of gestation. Information on prenatal supplementation with iron or calcium was acquired, and adverse perinatal outcomes were recorded. Biosamples from all 114 mothers and 73 newborns (61 singleton and 12 twin newborns) were finally analyzed. Total iron and calcium concentrations in serum were measured through total reflection X-ray fluorescence analysis. The results indicated no significant differences in maternal serum iron and calcium concentrations between singleton and twin pregnancies. Similarly, iron and calcium concentrations in newborn umbilical cord serum samples were not different between singleton and twin pregnancies. The comparison of total iron and calcium between mothers and umbilical cord serum indicated significantly lower concentrations in the mothers, with the differences being not homogenous but rather pair-specific. A significant positive correlation between maternal serum and umbilical cord serum calcium concentration was noticed. Prenatal iron supplementation was associated with higher iron concentrations in both mothers and newborns, supporting the efficiency of supplementation and the quality of the study methods. Collectively, the data indicate no significant differences in serum iron and calcium concentrations with regard to singleton or twin pregnancies and the efficiency of iron supplementation during pregnancy for increasing iron status.

The role of phoenixin in the proliferation and migration of ectopic epithelial cells in vitro.

AIM: Endometriosis is one of the most common gynecologic diseases in women of reproductive age. The pathophysiology of endometriosis is still not fully understood. Phoenixin (PNX-14) is a newly discovered neuropeptide that regulates the hypothalamo-pituitary-gonadal (HPG) axis and reproductive functions. Recently, we reported that PNX-14, its precursor protein and receptor were expressed in human endometrium. Moreover, PNX-14 serum levels in endometriosis were reduced. This study aimed to evaluate the in vitro biological functions of physiological PNX-14 concentrations on the ectopic endometrium Z12 cells. METHODS: The proliferation and migration of Z12 cells were assessed using the xCELLigence® RTCA DP system following 72 h of stimulation with 0.05 and 0.2 nM of PNX-14. GPR173 and small integral membrane protein 20 (SMIM20) gene expression was evaluated using quantitative polymerase chain reaction (qPCR) and the protein levels of GPR173 were analyzed using Western blot analysis. RESULTS: PNX-14 at the concentration observed in the serum of patients with endometriosis (0.05 nM) reduced GPR173 and increased SMIM20 expression, while protein levels of GPR173 remained unchanged. Cell proliferation was increased by the 0.02 nM PNX-14- the concentration found in healthy subjects. The 0.2 nM of PNX-14 decreased SMIM20 expression with no change to GPR173 expression and reduced ectopic epithelial cell proliferation during the first 5 h after stimulation. However, at 72 h, the proliferation increased. CONCLUSIONS: This study shows that PNX-14 at endometriosis specific concentration desensitized ectopic epithelium to PNX-14, and increased the expression of SMIM20 to restore the physiological levels of PNX-14.

The clinical significance of electronic fetal heart rate monitoring in twins.

OBJECTIVES: Fully effective intrapartum cardiotocographic (CTG) fetal heart monitoring is still missing. Visual analysis is far from credibility. Additional, computerized analysis techniques were proposed however they did not substantially decrease possible risks of fetal asphyxia. In twin pregnancies the problem is even more complicated. Our goal is to find the most valuable parameters in intrapartum CTG surveillance in twins, based on actual FIGO criteria. MATERIAL AND METHODS: Study included 58 women in labor who had been admitted to Delivery Department of tertiary care hospital with twin pregnancy in a period of one year. The features of the CTG (e.g., baseline, oscillation, decelerations, brady- or tachycardia) were grouped to create three variables that were closest to the FIGO CTG scale. All three groups were compared according to neonatal status (Apgar score at 5 min ≥ 7 or < 7; pH value in umbilical artery ≥ 7.20, < 7.20 or < 7.10 and BE (base excess) > or ≤ -12). Fetal status and its acid - base equilibrium was compared either with long term variability (LTV), short term variability (STV), or percentage of the signal loss. RESULTS: Out of 58 twin pregnancies, a total of 116 babies were born. One baby was born dead. From this group, 11 deliveries were natural births and 47 deliveries were C-sections. None of the analyzed features (pH, BE, Apgar, CTG features except tracing length, CTG FIGO categories) were statistically different between groups of singleton and twin pregnancies, except percentage of C-sections. No differences were found either for STV or LTV and fetal status.org CTG categories. CONCLUSIONS: Prior to cardiotocographic tracing of twins during labor, ultrasound examination should be mandatory. Considerable loss of signal in CTG tracing in twins should provoke ultrasonographic confirmation of the fetal status.

Differences in sex hormone levels in the menstrual cycle due to tobacco smoking - myth or reality?

INTRODUCTION: Tobacco smoke contains, among others, polycyclic aromatic hydrocarbons (PAHs), heterocyclic analogues, aromatic amines, N-nitrosamines, volatile hydrocarbons, aldehydes, phenols, miscellaneous organic compounds, metals, and inorganic compounds. Tobacco smoking can harm women's reproductive system and may reduce fertility. The objective of the study was to explore the effect of tobacco smoke on the menstrual cycle due to smoking and second-hand smoke-exposure. MATERIAL AND METHODS: The study was performed on 153 women of reproductive age, who received care at the Gynaecological-Obstetric Clinical Hospital of the Poznan University of Medical Sciences. They were divided into three treatment groups: non-smokers, secondhand smokers, and smokers. Comprehensive assessment of all hormone levels: follicle-stimulating hormone (FSH), luteinizing hormone (LH), 17ß-oestradiol (E2), and progesterone (P), in the various phases of the menstrual cycle and with concomitant determinations of serum cotinine concentrations was performed. The menstrual cycle was observed with ultrasonography. RESULTS: Cigarette smoking may be an important factor in disrupting reproduction: 1. The increase in the oestradiol E2 level was accompanied by significantly lowered serum cotinine concentrations in tobacco smokers; 2. In smoking patients, the serum level of LH significantly increased on the first days of the menstrual cycle; 3. The higher levels of P (in the 14th and 21st days) were assumed to be the result of a longer menstrual cycle. CONCLUSIONS: Active and passive smoking may be an important contributor to reproductive health issues and deserves greater focus in health education programs directed towards women of reproductive age.

Outcome dependent twin growth curves based on birth weight percentiles for Polish population.

OBJECTIVE: The objective of this study is to determine a healthy fetal growth pattern of twins from a Polish population based on an outcome-dependent growth curve. METHODS: The fetal growth data of live-born twin pregnancies between 25th and 40th week gestation in the period of 1 January 2005 to 31 March 2018 from the database of a tertiary care women's hospital in Western Poland was used to calculate birth weight percentiles. The growth curves of singletons from the same database were used as comparison. Because this study aimed for an outcome-dependent approach for the calculation of fetal growth curves, all babies born that may have high risk of unfavorable outcome were excluded. After application of all exclusion criteria, 1317 records referring to 2634 children were included in our analysis. Growth curves of singletons from the same database were used as reference for this study. RESULTS: A linear relationship between 10th, 50th, and 90th percentiles and gestational age were found for twins but not for singletons suggesting the different mechanisms of intrauterine growth between singleton and twin pregnancies. Week-to-week weight gain equal to or higher than 150 g in twins also predict a favorable outcome in absence of other pathologies. CONCLUSION: The calculated outcome-dependent fetal growth curves for twins in this study may help in the accurate diagnosis of small or large twin fetuses for their gestational age in this Western Poland population.

Phoenixin as a New Target in the Development of Strategies for Endometriosis Diagnosis and Treatment.

Small integral membrane protein 20/phoenixin (SMIM20/PNX) and its receptor GPR173 (G Protein-Coupled Receptor 173) play a role in the regulation of the hypothalamic-pituitary-gonadal axis (HPG). The aim of the study was to determine PNX, FSH, LH, and 17ß-estradiol association in women with endometriosis, and the expression of SMIM20/PNX signaling via GPR173. Serum PNX, FSH, LH, and 17ß-estradiol concentrations were measured by enzyme and electrochemiluminescence immunoassay. SMIM20/PNX and GPR173 expression in the eutopic and ectopic endometrium was assessed by qPCR and immunohistochemistry. Reduced PNX level, increased LH/FSH ratio and elevated 17ß-estradiol concentration were found in patients with endometriosis. No differences in SMIM20 expression were observed between the studied endometria. GPR173 expression was lower in ectopic than in eutopic endometria. SMIM20 expression was mainly restricted to stroma. GPR173 was detected in some eutopic and ectopic stromal cells and in eutopic glandular epithelial cells. Discriminant analysis indicates the diagnostic relevance of PNX and LH/FSH ratio in patients with endometriosis. In women with endometriosis, reduced PNX levels and GPR173 expression may be responsible for HPG axis dysregulation. These new insights may contribute to a better understanding of the pathophysiology of endometriosis and provide the basis for a new strategy for diagnosis and treatment of endometriosis.

Effect of Protoberberine-Rich Fraction of Chelidonium majus L. on Endometriosis Regression.

Endometriosis is a gynecological disease defined by the presence of endometrial tissue outside the uterus. To date, the effective treatment of this disease is still based on invasive surgery or laparoscopy. Chelidonium majus L. (Papaveraceae) belongs to medicinal, latex-bearing plants. Extracts from the plant are a rich source of pharmacologically active agents. Protoberberine compounds derived from C. majus possess anticancer and antiproliferative activities. In the present study of a rat model of endometriosis, we investigated the influence of the plant protoberberine-rich fraction (BBR) obtained from the medicinal plant C. majus on the development of endometriosis. To understand of BBR therapeutic potential for endometriosis, metabolomics has been applied to study. BBR was prepared from an ethanolic extract of dry plants C. majus. Rats (n = 16) with confirmed endometriosis were treated with BBR administered orally (1 g/kg) for 14 days. Blood serum samples were collected from all of the animals and metabolites were studied using the NMR method. The metabolomic pattern was compared before and after the protoberberine treatment. The performed analysis showed significant changes in the concentrations of metabolites that are involved in energy homeostasis, including glucose, glutamine, and lactate. Histopathological studies showed no recurrence of endometriosis loci after treatment with BBR. The results of the study found that BBR treatment prevents the recurrence of endometriosis in rats. Moreover, metabolomics profiling can be applied to better understand the mechanisms of action of these protoberberine secondary plant metabolites. Our findings provide new insights into the pharmaceutical activity of natural protoberberine plant compounds.

Outcome dependent growth curves for singleton pregnancies based on birth weight of babies for Polish population.

OBJECTIVES: To create outcome dependent fetal growth curves and birth weight standards that can be analyzed for use in clinic specifically for Western European populations. MATERIAL AND METHODS: We conducted a retrospective study on fetal growth and birth weight trends from live birth singleton pregnancies between 2005 and 2018 at one of the largest tertiary gynecologic-obstetric hospitals in Poland. The inclusion criteria were at least 22 weeks of gestation at birth regardless of delivery mode (vaginal or C-section), no congenital anomalies diagnosed before and after delivery and an Apgar score of at least 7 in the first minute. The final sample had a total of 39,413 cases (18,562 girls and 20,851 boys). We presented 7 (for all fetuses in the 5th, 10th, 25th, 50th, 75th, 90th and 95th percentiles) and 6 (for boys and girls each at 10th, 50th and 90th percentiles) fetal growth curves between 25 and 40 weeks of gestation. Birth weight trends were obtained and analyzed from all babies in the 5th, 10th, 25th, 50th, 75th and 95th percentiles born between 22 to 42 weeks of gestation with also separate trends for boys and girls. RESULTS: The largest differences in fetal growth curves were observed in the 10th and 50th percentiles between 22 and 34 weeks of gestation. A decreasing fetal weight gain pattern was observed between 27 and 30 weeks and after 38 weeks of gestation, the decrease was more drastic in female. A significant increase from 2009 to 2017 was observed in the weight of 50th percentile babies born at or after 35 weeks. We found significant discrepancies between our results and the most used European fetal growth curves particularly in the 10th and 90th percentile weights at 30 weeks. CONCLUSIONS: Separate scales for boys and girls were implied and given the overall difference form commonly used references. We believe there is significant value in using these unique patterns found in fetal growth curves and birth weights of ethnically homogenous population (such as Poland) at everyday clinical practice for more opportunities of safe obstetric care and higher chances of delivering a healthy child.

Threatened preterm labour - analysis of the cytokine profile and progesterone treatment efficiency.

OBJECTIVE: Preterm labour is defined as the onset of labour between weeks 23 and before 37 of gestation. The purpose of this study was to compare the cytokine profile in the blood serum of patients experiencing threatened premature labour, along with the time of delivery and the foetus body mass at birth. METHODS: The study included 89 women hospitalised at the Obstetrics and Gynaecology Clinical Hospital in the period 2011-2013. Group 1 comprised 31 patients diagnosed with threatened preterm labour, prior to tocolytic treatment. Group 2 comprised 32 patients with tocolytic treatment. Group 3 constituted the control group. RESULTS: Groups 1, 2 and 3 differed considerably in terms of the week of delivery of pregnancy (p = 0.006). Analogous results were obtained by analysing the body mass at birth, where a statistically significant difference in body mass at birth was found between Groups 1, 2 and 3 (p < 0.001). Our analysis focussed on the cytokine profile of the women included in the study, but no considerable changes in cytokine concentration were observed. CONCLUSIONS: No statistically significant differences were found in the cytokine profile among those patients in normally progressing pregnancy or in threatened preterm labour. No suitability of progesterone treatment in threatened preterm labour was observed.

Matrix metalloproteinase-2 (MMP-2), MMP-9, tissue inhibitor of matrix metalloproteinases (TIMP-1) and transforming growth factor-ß2 (TGF-ß2) expression in eutopic endometrium of women with peritoneal endometriosis.

INTRODUCTION: The prevalence of endometriosis among reproductive age women is 7-17%; however, these figures reach 20-50% in patients suffering from infertility. Matrix metalloproteinases (MMPs) activity is thought to be particularly essential in the early phases of endometriosis development. Any changes in the equilibrium between MMPs activity and their tissue inhibitors (TIMPs) could be potentially harmful, promoting endometriosis development. The aim of this study was to investigate whether the MMP-2, MMP-9, TIMP-1 or TGF-B2 expression in eutopic endometrium from women with early endometriosis differ when compared with healthy subjects. The results were referred to the serum progesterone levels. MATERIALS AND METHOD: Endometrial biopsy was taken from 42 patients (18 in the study group, 22 in thecontrol group) at the time of hysteroscopy for routine histology and for RT-PCR procedures. Comparison of the quantity of gene products was performed with a programme for densitometry and compared to GADPH product, which was a reference value. RESULTS: The obtained results did not reveal any statistical difference in endometrial expression of MMP-2, MMp-9, TIMP-1, and TGF-ß2 or serum progesterone level between women with endometriosis and without visible signs of this illness. CONCLUSION: Despite the lack of statistical differences, it was observed that both examined metalloproteinases expressed a tendency to higher gene expression in the eutopic endometrium of women with endometriosis. However, both TIMP-1 and TGF-ß2 expressions had the same tendency - higher values in endometriosis patients.

Selection of reliable reference genes in eutopic and ectopic endometrium for quantitative expression studies.

PURPOSE: Physiological changes during menstrual cycle cause the endometrium and endometriosis to develop specific kind of tissues, especially in regard to the gene expression profiles, which may include also housekeeping genes, commonly used as reference genes (RGs) in quantitative studies. Reverse transcription, followed by quantitative polymerase chain reaction (RT-qPCR) is the most precise and commonly used method in gene expression studies. In order to reduce effects of technical approaches and biological variability of gene's expression level, the studies often employ RGs in experimental data normalization. However, the expression of RGs is not always stable and depends on several variables. Thus, the selection of appropriate RG is one of the most significant steps to obtain reliable results in RT-qPCR-based methods. MATERIAL AND METHODS: With the usage of RT-qPCR, we researched the expression of seven genes (ACTB, B2M, G6PD, GAPD, GUSB, HPRT and PPIA) as reliable reference genes in eutopic and ectopic endometrial tissue specimens obtained during standard surgery of women of reproductive age. Stability of expression level was analyzed by the most universal MS Excel plug-ins including: geNorm, NormFinder and BestKeeper. The descriptive statistics were evaluated using Statistica software. RESULTS: The distribution of threshold (Ct) values was not equal. We identified genes with higher expression level (referring to Ct values) such as ACTB and B2M, medium e.g., GAPD and low expression level, e.g., G6PD and HPRT. We demonstrated that the stability of the analyzed reference genes was not homogenous, and different algorithms pointed to PPIA, GAPD and B2M as the most stable ones in eutopic and ectopic endometrium. On the contrary to these, GUSB and G6PD were the most unstable ones. CONCLUSIONS: In RT-qPCR-based analyses of gene expression level in eutopic and ectopic endometrium, we strongly recommend that a minimum of two reference genes are to be used and we determined that the most suitable seem to be PPIA and GAPD.

Recurrent fetal complex ovarian cysts with rupture followed by simple cyst in the neonatal period with no adverse sequelae.

Fetal ovarian cysts are the most frequent type of abdominal tumors in female fetuses with prenatal detection rate of more than 30%. The etiology of fetal ovarian cysts is unclear, but hormonal stimulation as well as presence of maternal diabetes, hypothyroidism, Rh iso-immune hemolytic disease and toxemia has been generally considered responsible for the disease. Complications of fetal ovarian cysts include compression of other viscera, cyst rupture, hemorrhage and, most frequently, ovarian torsion with consequent loss of the ovary. Management is controversial with several options described in the literature, including watchful expectancy, antenatal aspiration of simple cysts to prevent torsion and ovarian loss and finally, resection of all complex cysts in the neonatal period. To date, no case report has described recurrent complex cysts with rupture in the fetal period and recurrence of simple cyst in neonatal period. By presenting this case, we wanted to show that surgical intervention in case of prenatally diagnosed fetal ovarian cyst should be considered postnatally and only in symptomatic or complicated cases.

Prenatal diagnosis of vasa previa.

Vasa previa is a rare condition in which unsupported by the placenta, umbilical cord blood vessels runs within the placental membranes between internal os of the cervix and presenting part of the fetus. We report an antenatal diagnostic procedure and management of a patient with low-lying placenta and velamentous cord insertion near to the internal os with two large fetal blood vessels coursing between the internal cervical os and fetal presenting part. An elective cesarean section was performed at 36 weeks gestation.

[Effects of active and passive smoking during pregnancy on the development of gestational hypertension and fetal hypotrophy]. / Wplyw czynnego i biernego palenia papierosów przez kobiety ciezarne na rozwój nadcisnienia tetniczego w ciazy i hipotrofii u plodu.

In prenatal life the fetus can be exposed to more incentires which affect the mother. In case of both active and passive smoking by pregnant women many pernicious substances contained in tobacco smoke can influence the foetus. The components of tobacco smoke can contribute to various antenatal complications. The aim of present work was to assess if the data given by the patient in anonymous questionnaire go along with the real exposure to components of smoke assessed on the basis of concentration of nicotine metabolite- cotinine in mother's serum and next to assess the risk of development of gestational hypertension and hypotrophy. The biological material was extracted with means of liquid-liquid technique and next we performed laboratory determinations with means of liquid chro- matography with sectrophotometric detection, in which norephedrine is used as inner sample. The study was carried out on 106 women in perinatal period who give birth from single, full term pregnancy. On the basis of questionnaire data considering the concentration of cotinine, the patients were divided into three groups: I group--active smokers (n = 27), II group--passive smokers (n = 32), III group--not exposed to tobacco smoke (n = 41). The average concentration of cotinine in blood serum in the first group was 129.6 ng/ml, second group--5.1 ng/ ml, but in all patients from the control group the concentration of cotinine was below the detection level. Gestational hypertension was diagnosed in 17 women (16%) and hypotrophy of the foetus was diagnosed in 15 patients (14%). Tobacco smoking didn't influence the development of gestational hypertension and hypotrophy showered it is more often observed in women exposed to components of tobacco smoke during pregnancy.