Uterine smooth muscle tumors with uncertain malignant potential: analysis following fertility-saving procedures.

OBJECTIVE: The aim of this study was to analyze the clinical and reproductive outcomes of patients treated with myomectomy who were histologically diagnosed with uterine smooth muscle tumor of uncertain malignant potential (STUMP). METHODS: Patients who were diagnosed with STUMP and underwent a myomectomy at our institution between October 2003 and October 2019 were identified. Variables of interest obtained from the institution's database included patient age, relevant medical history, pre-operative appearance of the tumor on ultrasound, parameters of the surgical procedure, histopathological analysis of the tumor, post-operative clinical course, and course of follow-up, including reinterventions and fertility outcomes. RESULTS: There were a total of 46 patients that fulfilled the criteria of STUMP. The median patient age was 36 years (range, 18-48 years) and the mean follow-up was 47.6 months (range, 7-149 months). Thirty-four patients underwent primary laparoscopic procedures. Power morcellation was used for specimen extraction in 19 cases (55.9% of laparoscopic procedures). Endobag retrieval was used in nine patients and six procedures were converted to an open approach due to the suspicious peri-operative appearance of the tumor. Five patients underwent elective laparotomy due to the size and/or number of tumors; three patients had vaginal myomectomy; two patients had the tumor removed during planned cesarean section; and two underwent hysteroscopic resection.There were 13 reinterventions (five myomectomies and eight hysterectomies) with benign histology in 11 cases and STUMP histology in two cases (4.3% of all patients). We did not observe any recurrence as leiomyosarcoma or other uterine malignancy. We did not observe any deaths related to the diagnosis. Twenty-two pregnancies were recorded among 17 women, which resulted in 18 uncomplicated deliveries (17 by cesarean section and one vaginal), two missed abortions, and two pregnancy terminations. CONCLUSIONS: Our study found that uterus-saving procedures and fertility-preservation strategies in women with STUMP are feasible, safe, and seem to be associated with a low risk of malignant recurrence, even while maintaining the mini-invasive laparoscopic approach.

Periphyton (microbiome) of filamentous algae studied by polarization microscopy, aided also by some contrasting optical methods.

Polarization microscopy, possibly together with some contrast techniques (dark field and color phase contrast), was used to study the periphyton (microbiome) growing on filamentous green algae. The material containing filamentous algae with periphyton on the surface was collected in the villages of Sýkorice and Zbecno (Krivoklátsko Protected Landscape Area). The objects were studied in a LOMO MIN-8 St. Petersburg polarizing microscope and a Carl Zeiss Jena NfpK laboratory microscope equipped with an In Ph 160 basic body with variable dark field or color phase contrast and a Nikon D70 DSLR digital camera. Cells of filamentous algae of the genera Cladophora, Vaucheria, and Oedogonium were studied and the periphyton attached to them formed by cyanobacteria of the genera Chamaesiphon and Pleurocapsa and algae of the genera Characium, including diatoms of the genera Eunotia and Synedra. In all cases, the cell walls of the host algae showed a very strong birefringence. In contrast, the walls of cyanobacteria of the genera Chamaesiphon and Pleurocapsa were characterized by a much weaker birefringence (Pleurocapsa somewhat thicker), and the diatom frustules of the genera Eunotia and Synedra were almost without a birefringence. Strongly birefringent granules were found in the cytoplasm of the green alga of the genus Characium, which forms periphyton on the filamentous green algae of the genus Vaucheria. The periphyton on the filamentous alga of the genus Oedogonium, formed by cyanobacteria of the genus Pleurocapsa and diatoms of the genera Eunotia and Synedra, deposited in a massive layer of mucus containing birefringent crystals, showed a particularly strong birefringence. At the end of the vegetation of filamentous algae, their parts and remnants of periphyton (diatom frustules and crystals) became part of the detritus at the bottom of the culture vessel. The use of polarization microscopy in the study of filamentous algae with periphyton on the surface allows us not only to determine the birefringence of the observed structures, but also to partially deduce their chemical composition, or regular arrangement of particles, so-called shape birefringence.

Hysteroscopic findings after laparoscopic and open myomectomy with or without uterine cavity breach: historical cohort study.

INTRODUCTION: The aim of this study was to evaluate hysteroscopic findings after laparoscopic and laparotomic myomectomy with a focus on the presence of postoperative intrauterine adhesions in groups of patients with and without perioperative uterine cavity breach (UCB). MATERIAL AND METHODS: This is a historical cohort study. Our database was searched to identify patients with UCB during myomectomy and matched the same number of patients after myomectomy without UCB to create a control group. All relevant data were retrieved from our medical records. In both groups, the results of follow-up hysteroscopy were analyzed. RESULTS: Low prevalence of intrauterine adhesions after myomectomy was observed in only 3.5% of the 170 patients in our samples. No significant difference in the occurrence of synechiae between the patients with and without UCB was found (2 vs. 4, RR 0.5, 95% CI 0.1-2.7, p = .341), nor was the difference in other hysteroscopic findings. Follow-up hysteroscopy was performed with slender optics and expandable casing system without need of any anesthesia in 87.1% of cases. CONCLUSIONS: According to our findings, the prevalence of post-myomectomy intrauterine adhesions after myomectomy is low. Our study did not demonstrate that UCB during myomectomy is a risk factor for the formation of intrauterine synechiae.

Hysteroscopic management of endometrial polyps and submucous leiomyomas in women using a levonorgestrel-releasing intrauterine system.

INTRODUCTION: This study was designed to evaluate the feasibility and effectiveness of hysteroscopy in the management of symptoms related to endometrial polyps and submucous leiomyomas in women using a levonorgestrel-releasing intrauterine system (LNG-IUS). MATERIAL AND METHODS: Twenty-three LNG-IUS users presenting with endometrial polyps and/or submucous leiomyomas and irregular uterine bleeding were recruited for hysteroscopic examination and surgery. Intrauterine pathology was investigated and treated by hysteroscopic resection with the LNG-IUS in situ, and the effect of the procedure on symptoms was evaluated after three to six months. RESULTS: Intrauterine pathology was successfully resected by hysteroscopy in 23 (100.0%) out of 23 cases. Following hysteroscopy, 18 (78.3%) women reported amenorrhea, one (4.3%) regular spotting, three (13.0%) irregular spotting and one (4.3%) patient resumed normal menstrual cycle. We conclude that 19 (82.6%) patients were postoperatively asymptomatic. All procedures were uncomplicated and 4 (17.4%) were carried out without general anesthesia as office procedures. CONCLUSION: Endometrial polyps and submucous leiomyomas can develop in LNG-IUS users, and this can cause irregular uterine bleeding. Hysteroscopic resection of these pathologies is a feasible method in the clinical management of symptoms.

Simultaneous use of polarization microscopy and dark field techniques in the study of microorganisms.

Simultaneous application of polarization microscopy and dark field techniques has been used to study the internal structure of microbial cells. The dark field technique displays subtle cell structures like glowing objects on a dark background. In the polarizing microscope, cross polarizing filters along with the first-order quartz compensator and a rotary table show the maximum birefringence of the individual structures. The material containing microorganisms was collected in the villages of Sýkorice and Zbecno (Krivoklátsko Protected Landscape Area). The objects were studied in a laboratory microscope Carl Zeiss Jena type NfpK equipped with In Ph 160 basic body with variable dark field, special condenser with interchangeable diaphragm apertures, a rotary table, Meopta Praha polarizer, analyzer, first-order quartz compensator from LOMO Sankt Petersburg, and a digital Nikon D 70 DSLR camera. Three orders of microorganisms were studied: Siphonocladales, Chlorococcales, and Peritricha. Anisotropic structures in different amounts and sizes (e.g., granules and microfibrils) or in different configurations (e.g., cell walls or pellicle) have been found in all Protista organisms under study. Filamentous algae of the genus Cladophora (Cladophoraceae, Siphonocladales, Ulvophyceae) featured a strongly birefringent cell wall (shape birefringence) surrounded by less birefringent periphyton (microbial biofilm), at the edges of which cyanobacterial fibers could be recognized-a very important finding. The coccal algae of the genus Scenedesmus (Scenedesmataceae, Chlorococcales, Chlorophyceae) exhibited not only strongly birefringent granules, but also strongly birefringent microfibrils in the cytoplasm outside the strongly birefringent cell walls-very important finding. Of all the studied microorganisms, the weakest birefringence was shown in the surface membrane (pellicle) of the Vorticella (Vorticellidae, Peritricha, Ciliata). On the other hand, the ring of cilia on the top of the body had a somewhat stronger birefringence-an important finding. In conclusion, the dark field technique provides a high contrast image in the microscope and, if supplemented simultaneously by polarization microscopy, will allow us to partially infer the composition of the examined structures.

Simultaneous use of Interphako interference contrast and polarization microscopy in the study of microorganisms.

Simultaneous application of polarization microscopy and Interphako interference contrast has been used to study the internal structure of algal cells. The interference contrast technique showed fine cell structures (important is the selection of interference colors according to the Mach-Zehnder interferometer setting). In a polarization microscope, the crossed polarization filters together with the first-order quartz compensator mounted turntable showed the maximum birefringence of the individual structures. Material containing green algae was collected in the villages Sýkorice and Zbecno, Protected Landscape Area (PLA) Krivoklátsko. The objects were studied in a Carl Zeiss Jena NfpK laboratory microscope equipped with an In 160 base body with an Interphako In contrast interference module including a Mach-Zehnder interferometer with variable phase contrast, a special condenser with interchangeable aperture plates, a turntable, a Meopta Praha polarizer, a LOMO Sankt Petersburg analyzer, and a quartz compensator with first-order red and the digital camera DSLR Nikon D 70. Green algae of three orders were studied: Siphonocladales, Zygnematales, and Desmidiales. Anisotropic structures were found in all studied representatives of the green algae of the phylum Chlorophyta. Especially their cell walls showed strong birefringence (in all representatives of these orders). On the other hand, a representative of the order Siphonocladales (the genus Cladophora, Cladophoraceae, Ulvophyceae) was rarely found to display weak birefringent granules of storage substances due to the setting of the Mach-Zehnder interferometer and the use of the first-order compensator (interference colors are intensified). In addition, a very weak birefringence of periphyton cells (microbial biofilm) was found. In the study of the second algae of the genus Spirogyra (Zygnemataceae, Zygnematales, Conjugatophyceae), a strongly birefringent connecting wall between algal cells was found in contrast to the weaker birefringence of the peripheral wall. It was the use of Interphako interference contrast together with polarization filters and a first-order quartz compensator that particularly emphasized the central part of the connecting wall. In the study of the twinned Pleurotaenium algae (Desmidiaceae, Desmidiales, Conjugatophyceae), a strongly birefringent wall was found along the periphery of the cell with a nucleus in the middle part (isthmus). In this narrowing in the center of the cell, a sharply delimited birefringent edge of the cell wall is visible, especially when using Interphako interference contrast along with crossed polarization filters and a first-order quartz compensator. In conclusion, Interphako interference contrast provides a high degree of image contrast in a microscope and, if suitably simultaneously complemented by polarization microscopy (including a first-order quartz compensator), it will allow us to infer some of the composition of the investigated structures. However, working with Interphako interference contrast is considerably more difficult (setting Mach-Zehnder interferometer) than using other contrast techniques (positive and negative phase contrast, color contrast, relief contrast, and dark field).

Malignant endometrial polyp in woman with the levonorgestrel intrauterine system - a case report.

The levonorgestrel-releasing intrauterine system (LNG-IUS) is widely used and recommended as a reliable contraceptive. It also acts by opposing the effects of estrogen on the endometrium, thereby preventing development of endometrial hyperplasia and its possible malignant transformation. This case describes a 52-year-old multiparous amenorrhoeic patient who was seen in the gynecology outpatient department for a routine control 46 months after the insertion LNG-IUS as contraception. Hysteroscopy with a target biopsy following suspicious ultrasound scan confirmed well-differentiated endometrioid adenocarcinoma. Ultrasound scan prior to inserting LNG-IUS revealed normal 5 mm thin endometrium with the sharp edges. Uterine bleeding before the LNG-IUS insertion was regular and not excessive and the woman has remained amenorrhoeic after the LNG-IUS insertion. We present a case of the growth of a polyp-shaped endometrial carcinoma in a LNG-IUS asymptomatic user.

Concomitant use of polarization and negative phase contrast microscopy for the study of microorganisms.

A simultaneous application of negative phase contrast and polarization microscopy was used to study the internal structure of microbial cells. Negative phase contrast allowed us to display the fine cell structures with a refractive index of light approaching that of the environment, e.g., the cytoplasm, and converted an invisible phase image to a visible amplitude one. In the polarizing microscope, cross-polarizing filters, together with first-order quartz compensator and a turntable, showed maximum birefringence of individual structures. Material containing algae was collected in ponds in the villages Sýkorice and Zbecno (Protected Landscape Area Krivoklátsko). Objects were studied in a laboratory microscope (Carl Zeiss Jena, type NfpK), equipped with a basic body In Ph 160 with an exchangeable module Ph, LOMO St. Petersburg turntable mounted on a centering holder of our own construction and a Nikon D 70 digital SLR camera. Anisotropic granules were found only in the members of two orders of algae (Euglenales, Euglenophyceae and Chlorococcales, Chlorophyceae). They always showed strong birefringence and differed in both number and size. An important finding concerned thin pellicles in genus Euglena (Euglenales, Euglenophyceae) which exhibited weak birefringence. In genus Pediastrum (Chlorococcales, Chlorophyceae), these granules were found only in living coenobium cells. In contrast, dead coenobium cells contained many granules without birefringence-an important finding. Another important finding included birefringent lamellar structure of the transverse cell wall and weak birefringence of pyrenoids in filamentous algae of genus Spirogyra (Zygnematales, Conjugatophyceae). It was clearly displayed by the negative phase contrast and has not been documented by other methods. This method can also record the very weak birefringence of the frustule of a diatom of genus Pinnularia (Naviculales, Bacillariophyceae), which was further reinforced by the use of quartz compensator-an important finding. Simultaneous use of negative phase contrast and polarization microscopy allowed us to study not only birefringent granules of storage substances in microorganisms, but also the individual lamellae of the cell walls of filamentous algae and very thin frustule walls in diatoms. These can be visualized only by this contrast method, which provides a higher resolution (subjective opinion only) than other methods such as positive phase contrast or relief contrast.

Occurrence of Microsporidium sp. and other pathogens in Ips amitinus (Coleoptera: Curculionidae).

A new microsporidium is reported from the small spruce bark beetle, Ips amitinus: Microsporidium sp. with uninucleate oval spores measuring 3.5 × 2.5 µm; infecting cells of the midgut epithelium, midgut muscles, the fat body, the Malpighian tubules, and the gonads of adult beetles collected in Austria. Seven other pathogens were found in beetles collected from Austria, the Czech Republic, and Finland. Six of them were already known from I. amitinus. Nosema cf. typographi is recorded for the first time in the overwintering generation of I. amitinus from the Czech Republic.

Impact of maternal diabetes type 1 on proliferative potential, differentiation and apoptotic activity in villous capillaries of term placenta.

INTRODUCTION: Maternal diabetes mellitus changes morphology and impairs function of placental capillaries. Here, quantitative parameters characterizing cell proliferation using detection of Ki67, differentiation reflected by nestin expression and apoptosis in placental capillary bed with active caspase 3 as a marker were compared in normal term placentas and placentas from pregnancies complicated by Type 1 maternal diabetes mellitus. METHODS: Specimens of sixteen diabetic placentas and eight control placentas were collected by systematic uniform random sampling. Immunohistochemical detections of Ki67, nestin, and active caspase 3 were performed in histological sections of five haphazardly chosen blocks per placenta. Twenty fields of view per section, i.e. one hundred fields of view per placenta, were used for analysis of proliferation as well as of apoptosis, and in approximately 70 capillary cross-sections per placenta the nestin-positive segments of their circumference were measured. RESULTS: The percentage of Ki67-positive cells counted in the capillary wall was significantly lower in diabetic group. The counts of Ki67-labelled nuclei per villous area unit were significantly lower in cytotrophoblast and capillary wall of terminal villi in diabetic placenta. The proportion of nestin-labeled segments of capillary circumference was significantly higher in placentas of diabetic group. No differences in the numbers of apoptotic cells were found between studied groups. DISCUSSION: The results show that the term placenta in Type 1 diabetes has lower potential to enlarge the surface area of structures involved in maternofetal transport, and that the villous capillary bed displays delayed differentiation. Those factors may participate in decreased ability of diabetic placenta to comply with fetal requirements in the final stage of pregnancy.

Effect of metal ions on autofluorescence of the dry rot fungus Serpula lacrymans grown on spruce wood.

This work describes autofluorescence of the mycelium of the dry rot fungus Serpula lacrymans grown on spruce wood blocks impregnated with various metals. Live mycelium, as opposed to dead mycelium, exhibited yellow autofluorescence upon blue excitation, blue fluorescence with ultraviolet (UV) excitation, orange-red and light-blue fluorescence with violet excitation, and red fluorescence with green excitation. Distinctive autofluorescence was observed in the fungal cell wall and in granula localized in the cytoplasm. In dead mycelium, the intensity of autofluorescence decreased and the signal was diffused throughout the cytoplasm. Metal treatment affected both the color and intensity of autofluorescence and also the morphology of the mycelium. The strongest yellow signal was observed with blue excitation in Cd-treated samples, in conjunction with increased branching and the formation of mycelial loops and protrusions. For the first time, we describe pink autofluorescence that was observed in Mn-, Zn-, and Cu-treated samples with UV, violet or. blue excitation. The lowest signals were obtained in Cu- and Fe-treated samples. Chitin, an important part of the fungal cell wall exhibited intensive primary fluorescence with UV, violet, blue, and green excitation.

Concomitant use of polarization and positive phase contrast microscopy for the study of microbial cells.

Polarization and positive phase contrast microscope were concomitantly used in the study of the internal structure of microbial cells. Positive phase contrast allowed us to view even the fine cell structure with a refractive index approaching that of the surrounding environment, e.g., the cytoplasm, and transferred the invisible phase image to a visible amplitude image. With polarization microscopy, crossed polarizing filters together with compensators and a rotary stage showed the birefringence of different cell structures. Material containing algae was collected in ponds in Sýkorice and Zbecno villages (Krivoklát region). The objects were studied in laboratory microscopes LOMO MIN-8 Sankt Petersburg and Polmi A Carl Zeiss Jena fitted with special optics for positive phase contrast, polarizers, analyzers, compensators, rotary stages, and digital SLR camera Nikon D 70 for image capture. Anisotropic granules were found in the cells of flagellates of the order Euglenales, in green algae of the orders Chlorococcales and Chlorellales, and in desmid algae of the order Desmidiales. The cell walls of filamentous algae of the orders Zygnematales and Ulotrichales were found to exhibit significant birefringence; in addition, relatively small amounts of small granules were found in the cytoplasm. A typical shape-related birefringence of the cylindrical walls and the septa between the cells differed in intensity, which was especially apparent when using a Zeiss compensator RI-c during its successive double setting. In conclusion, the anisotropic granules found in the investigated algae mostly showed strong birefringence and varied in number, size, and location of the cells. Representatives of the order Chlorococcales contained the highest number of granules per cell, and the size of these granules was almost double than that of the other monitored microorganisms. Very strong birefringence was exhibited by cell walls of filamentous algae; it differed in the intensity between the cylindrical peripheral wall and the partitions between the cells. Positive phase contrast enabled us to study the morphological relationship of various fine structures in the cell (poorly visible in conventional microscope) to anisotropic structures that have been well defined by polarization microscopy.

Anisotropic structures of some microorganisms studied by polarization microscopy.

Polarization microscopy has been used to study the internal structures of microbial cells and in terms of the birefringence of these structures and its possible relation to the cell function and composition. Cyanobacteria of the genus Phormidium were found to contain no anisotropic structures, while other microorganisms were found to contain them, albeit to a different extent, size, and number. The flagellate Euglena was found to contain two large anisotropic bodies, whereas the flagellate of the genus Phacus belonging to the same systematic group Euglenales was observed to contain only one large anisotropic body (storage substances--paramylon). On the other hand, green algae of the genus Scenedesmus, whose cells form four--celled coenobia, contained clusters of small anisotropic granules composed also of storage substances (volutin). Minute anisotropic granules (storage substances) in two smaller clusters were found also in diatoms of the genus Navicula, whereas the green alga of the genus Mougeotia was revealed to contain, in addition to minute anisotropic granules (storage substances) occurring in low numbers in the cytoplasm, also a strongly birefringent cell wall (shape birefringence). Cells of the amoeba of the genus Naegleria and heliozoans of the genus Heterophrys were observed to contain only isolated tiny anisotropic granules (storage substances).

Steroid profiling in pregnancy: a focus on the human fetus.

In this review we focused on steroid metabolomics in human fetuses and newborns and its role in the physiology and pathophysiology of human pregnancy and subsequent stages of human life, and on the physiological relevance of steroids influencing the nervous systems with regards to their concentrations in the fetus. Steroid profiling provides valuable data for the diagnostics of diseases related to altered steroidogenesis in the fetal and maternal compartments and placenta. We outlined a potential use of steroid metabolomics for the prediction of reproductive disorders, misbalance of hypothalamic-pituitary-adrenal axis, and impaired insulin sensitivity in subsequent stages of human life. A possible role of steroids exhibiting a non-genomic effect in the development of gestational diabetes and in the neuroprotection via negative modulation of AMPA/kainate receptors was also indicated. Increasing progesterone synthesis and catabolism, declining production of tocolytic 5ß-pregnane steroids, and rising activities of steroid sulfotransferases with the approaching term may be of importance in sustaining pregnancy. An increasing trend was demonstrated with advancing gestation toward the production of ketones (and 3ß-hydroxyl groups in the case of 3α-hydroxy-steroids) was demonstrated in the fetus on the expense of 3α-hydroxy-, 17ß-hydroxy-, and 20α-hydroxy-groups weakening in the sequence C17, C3, and C20. There was higher production of active progestogen but lower production of active estrogen and GABAergic steroids with the approaching term. Rising activities of placental CYP19A1 and oxidative isoforms of HSD17B, and of fetal CYP3A7 with advancing gestation may protect the fetus from hyperestrogenization. This article is part of a Special Issue entitled 'Pregnancy and Steroids'.

Simple sets for digital microphotography used and tested in the study of microorganisms.

Four simple sets for digital microphotography are described that have been tested with the Carl Zeiss Jena, Meopta Prague, Lambda Prague, and LOMO Sankt Petersburg microscopes and with DSLR Nikon D 70 and Nikon D 300 cameras. They permit precise image focusing in the camera using a prism Zeiss. The sets make use of commonly available extensions Zeiss, Praktica and reductions Nikon-Praktica manufactured by ROWI (without a lens) or HAMA (with a lens). An extension has further been designed and manufactured for connecting the DSLR Nikon D 300 camera fitted with the HAMA reduction (only with a lens) and a focusing extensible prism with Zeiss Jena light measurement. It permits a precise image focusing of low light intensity objects (autofluorescence or low-contrast or moving objects when using positive or negative phase contrast). The sets are applicable to all microscopes constructed according to German DIN industrial standards.

Should hysteroscopy be provided for patients who have undergone instrumental intrauterine intervention after delivery?

We investigated the frequency of pathology, especially intrauterine adhesions, after instrumental evacuation within 24 h of delivery in a prospective observational intervention study on 100 women where a 'see and treat' hysteroscopy was performed after three to four months. There were two possible etiology groups: intrauterine adhesions [classified according by European Society for Gynaecological Endoscopy (ESGE) grades I-IV] and residual tissue (classified as minimal and considerable). Adhesions were found in 18% of patients, as follows: ESGE I-II in 13% and ESGE III-IV in 5%. Residual tissue was present in 33%, as follows: minimal in 23% and considerable in 10%. There were 6% who had both mild adhesions and minimal residual tissue, while 43% of the women had normal intrauterine findings. Of the women, 32% were symptomatic (spotting, bleeding). Only residual tissue correlated with symptoms (r=0.376; p<0.001). There is a high prevalence of acquired intrauterine pathology (57%) in women who require early instrumental evacuation.

Steroid metabolome in fetal and maternal body fluids in human late pregnancy.

Despite the extensive research during the last six decades the fundamental questions concerning the role of steroids in the initiation of human parturition and origin and function of some steroids in pregnancy were not definitely answered. Based on steroid metabolomic data found in the literature and our so far unpublished results, we attempted to bring new insights concerning the role of steroids in the sustaining and termination of human pregnancy, and predictive value of these substances for estimation of term. We also aimed to explain enigmas concerning the biosynthesis of progesterone and its bioactive catabolites considering the conjunctions between placental production of CRH, synthesis of bioactive steroids produced by fetal adrenal, localization of placental oxidoreductases and sustaining of human pregnancy. Evaluation of data available in the literature, including our recent findings as well as our new unpublished data indicates increasing progesterone synthesis and its concurrently increasing catabolism with approaching parturition, confirms declining production of pregnancy sustaining 5ß-pregnane steroids providing uterine quiescence in late pregnancy, increased sulfation of further neuroinhibiting and pregnancy sustaining steroids. In contrast to the established concept considering LDL cholesterol as the primary substrate for progesterone synthesis in pregnancy, our data demonstrates the functioning of alternative mechanism for progesterone synthesis, which is based on the utilization of fetal pregnenolone sulfate for progesterone production in placenta. Close relationships were found between localization of placental oxidoreductases and consistently higher levels of sex hormones, neuroactive steroids and their metabolites in the oxidized form in the fetus and in the reduced form in the maternal compartment.

Steroid metabolome in plasma from the umbilical artery, umbilical vein, maternal cubital vein and in amniotic fluid in normal and preterm labor.

The boost in placental production of CRH in late pregnancy is specific for human. CRH receptors are expressed in the fetal zone of the fetal adrenal (FZFA). Hence, we evaluated the associations between the steroid metabolome and gestational age (GA). The levels of 69 steroids and steroid polar conjugates such as 3beta-hydroxy-5-ene steroids (3betaOH5S), 3-oxo-4-ene steroids (3O4S), progesterone 5alpha/beta-reduced metabolites, 20alpha-hydroxy-metabolites of C21 steroids, C19 5alpha/beta-reduced metabolites, 7alpha/beta-hydroxy-metabolites of 3betaOH5S, estrogens and 16alpha-hydroxy-metabolites of 3betaOH5S and 3O4S, were measured by GC-MS in plasma from the umbilical artery (UA), umbilical vein (UV), and maternal cubital vein (MV) and in amniotic fluid (AF) in 12 women at normal labor and 38 women at preterm labor due to pathologies unrelated to steroid status. Using multivariate regression, prediction models for GA were completed for the individual body fluids. The conjugated 3betaOH5S (the key products of the FZFA), estrogens, some polar conjugates of progesterone 5alpha/beta-reduced metabolites and some steroid 7alpha/beta- and 16alpha-hydroxy-metabolites showed strong positive correlations with the GA. The predictivity decreased in the following sequence UV (R=0.950), UA (R=0.945), MV (R=0.895), and AF (R=0.891). Although the predictivity of steroids in maternal blood was slightly less effective when compared with the UV and UA, it was the best solution for further practice.

The distribution of placental oxidoreductase isoforms provides different milieus of steroids influencing pregnancy in the maternal and fetal compartment.

Using information based on the steroid metabolome in maternal and fetal body fluids, we attempted to ascertain whether there is a common mechanism, which is based on the placental distribution of various isoforms of 17ß-hydroxysteroid dehydrogenases and aldo-keto reductases. This system simultaneously provides a higher proportion of active progestogens in fetal circulation and a higher proportion of active estrogens and GABAergic steroids in the maternal compartment. The data obtained using gas chromatography-mass spectrometry completely support the aforementioned hypothesis. We confirmed a common trend to higher ratios of steroids with hydroxy-groups in the 3α-, 17ß-, and 20α-positions to the corresponding 3-oxo-, 17-oxo-, and 20-oxo-metabolites, respectively, in the maternal blood when compared with the fetal circulation, and the same tendency was obvious in the 3α-hydroxy/3ß-hydroxy steroid ratios. A decreasing trend was observed in the ratios of active estrogens and neuro-inhibitory steroids to their inactive counterparts in fetal and maternal body fluids. This was probably associated with a limited capacity of placental oxidoreductases in the converting of estrone to estradiol during the transplacental passage. Although we observed a decreasing trend in pregnancy-sustaining steroids with increasing gestational age, we recorded rising levels of estradiol and particularly of estriol, regardless of the limited capacity of placental oxidoreductases. Besides the estradiol, which is generally known as an active estrogen, estriol may be of importance for the termination of pregnancy with respect to its excessive concentrations near term which allows its binding to estrogen receptors.

ABO fetomaternal compatibility poses a risk for massive fetomaternal transplacental hemorrhage.

OBJECTIVE: Severe fetomaternal transplacental hemorrhage increases the risk of fetal anemia. In the third trimester, the syncytiotrophoblast becomes thinner, especially in areas where it comes into intimate contact with villous capillaries, and forms a vasculosyncytial membrane. Our aim was to determine whether ABO compatibility puts the fetus at a greater risk of severe fetomaternal hemorrhage. DESIGN: Case study. SETTING: A tertiary care center. Sample and methods. Between 2003 and 2007, we evaluated eight cases of severe fetomaternal transfusion. The Kleihauer-Betke test was used for diagnosis of fetomaternal hemorrhage. We evaluated blood group compatibility between the mother and fetus and assessed the perinatal outcome. The Fischer's factorial test was used for testing a hypothesis. RESULTS: The incidence of adverse outcomes following transplacental hemorrhage was 75% (six of eight). There were two perinatal deaths and four infants were affected by post-hypoxic damage of varying severity. Fetomaternal ABO compatibility was present in seven of the eight cases. The risk of severe fetomaternal hemorrhage was significantly increased when there was ABO compatibility between the mother and fetus. This was associated with a very poor perinatal outcome. CONCLUSION: We recommend that resuscitation in utero by intrauterine transfusion should be considered before the 33rd week of gestation in cases of severe fetal anemia. In later gestation, urgent cesarean section is required with adequate resuscitation of the newborn.