Verhalten der Gewichtsdifferenz von selektiv wachstumsretardierten, monochorialen Zwillingen

Problemstellung: Monochoriale Zwillinge (MZ) machen nur ca. 1/3 aller Mehrlinge aus. Im Vergleich zu dichorialen weisen sie jedoch eine höhere Morbidität und Mortalität auf bedingt durch das Transfusionssyndrom, mehr Fehlbildungen und Frühgeburten. In letzter Zeit kristallisiert sich eine weitere Entität heraus, die selektive intrauterine Wachstumsretardierung (sIUWR). Ziel der Studie war es, das Verhalten dieses diskordanten Wachstums im Verlaufe der Schwangerschaft zu untersuchen. Patienten und Methode: Die Diagnose einer monochorialen Situation basiert auf der Darstellung nur einer Plazenta, dem T-Sign sowie gleichgeschlechtiger Feten. Es wurden nur biamniote MZ eingeschlossen. Von einer sIUWR spricht man, wenn folgende Kriterien erfüllt sind: 1) ein Fetus wachstumsretardiert (Abdomenumfang <5. Perzentile) und 2) Gewichtsdifferenz beider Feten >20%. Letztere berechnet sich aus der Differenz der geschätzten fetalen Gewichte dividiert durch das Gewicht des Größeren in% (Delta%). Es wurden die Delta%-Werte bei Diagnosestellung und die letzten Werte vor Geburt verglichen. Ergebnisse: Retrospektiv wurden 23 Fälle mit sIUWR eingeschlossen, davon wurden 13 Fälle longitudinal beurteilt. Das mediane Gestationsalter bei Geburt lag bei 31,4 (16,7–38,6) Wochen. Die Frühgeburtsrate betrug 85,7%, die perinatale Mortalität 11,9%. Der initiale Delta%-Wert lag bei 28±10% und stieg signifikant auf 36,4±9,9%. In allen Fällen nahm der Delta%-Wert zu. Von den Co-Zwillingen waren postnatal 20% auch small for gestational age. Schlussfolgerungen: Bei MZ mit sIUWR ist nach unseren Erfahrungen damit zu rechnen, dass die Gewichtsdifferenz mit zunehmendem Gestationsalter steigt. Diese Zunahme erklärt sich durch eine progressive Verlangsamung der Wachstumsgeschwindigkeit des kleineren Feten. Leider lässt sich anhand der kleinen Fallzahl nicht eruieren, ob dieser D%-Wert und der Grad des Anstieges eine prognostische Bedeutung aufweist.

The umbilical cord in Cantrell: Pentalogy or Hexalogy?

Objective: Pentalogy of Cantrell (PC) is a rare congenital defect associated with five midline anomalies. The type of cardiac malformation and the size of the abdominal wall defect is often responsible for the high mortality. Of interest, the embryonic period in which PC develops is similar to that of the umbilical cord’s (UC) formation. The aim of the following study was to investigate the relationship between UC anomalies and PC. Methods: Charts of four cases with PC from 2002–08 were retrospectively reviewed for associated UC anomalies. UC anomalies were defined as single umbilical artery (SUA), short cord (during 1st trimester less than CRL or less than 30cm at term) or atypical UC coiling pattern. Results: We identified four cases: 3 singletons and one monochorionic diamniotic twin pregnancy with TRAP sequence. All cases showed a normal karyotype. All but one demonstrated the classical pulsatile omphalocele with ectopia cordis and all others anomalies of PC. One case was characterized by a major cranial omphalocele without ectopia cordis and no UC anomaly. This fetus was delivered by Cesarean at term and successfully operated on d1. In all other cases the parents requested ToP. Among the three cases with ectopia cordis, two had a short UC with SUA and one a short three-vessel cord; all these three UC were markedly uncoiled. Conclusions: Our data suggest a strong association between Cantrell and the development of the UC, in particular in cases with ectopia cordis. One might speculate that hemodynamic alterations of the feto-placental blood flow because of the cardiac malformation or structural changes at the umbilical ring (omphalocele) influence the development of the UC. More observations are needed to decide if Cantrell is a ‘‘hexalogy’’ instead of pentalogy.

Hyperaldosteronism in pregnancy

Aldosterone is a key regulator of electrolyte and water homeostasis and plays a central role in blood pressure regulation. Hormonal changes during pregnancy, among them increased progesterone and aldosterone production, lead to the required plasma volume expansion of the maternal body as an accommodation mechanism for fetus growth. This review discusses the regulation of aldosterone production by aldosterone synthase (CYP11B2); the impact on aldosterone secretion due to the presence of a chimeric gene originating from a crossover between CYP11B1 and CYP11B2 in glucocorticoid remediable aldosteronism (GRA) - the inherited form of hypertension; enhanced aldosterone production in aldosterone-producing adenoma (APA); and idiopathic hyperaldosteronism (IHA). Features of hyperaldosteronism are also found in patients with apparent mineralocorticoid excess (AME), in which glucocorticoids exacerbate activation of the mineralocorticoid receptor (MR) because of a defect in the 11beta-hydroxysteroid dehydrogenase type 2 enzyme. Regulation of aldosterone production and tissue-specific activation of the mineralocorticoid receptor are prerequisites for optimal control of body fluids and blood pressure during pregnancy and contribute largely to the wellbeing of the mother-to-be.

Cloning and characterization of two bistructural S-layer-RTX proteins from Campylobacter rectus

Campylobacter rectus is an important periodontal pathogen in humans. A surface-layer (S-layer) protein and a cytotoxic activity have been characterized and are thought to be its major virulence factors. The cytotoxic activity was suggested to be due to a pore-forming protein toxin belonging to the RTX (repeats in the structural toxins) family. In the present work, two closely related genes, csxA and csxB (for C. rectus S-layer and RTX protein) were cloned from C. rectus and characterized. The Csx proteins appear to be bifunctional and possess two structurally different domains. The N-terminal part shows similarity with S-layer protein, especially SapA and SapB of C. fetus and Crs of C. rectus. The C-terminal part comprising most of CsxA and CsxB is a domain with 48 and 59 glycine-rich canonical nonapeptide repeats, respectively, arranged in three blocks. Purified recombinant Csx peptides bind Ca2+. These are characteristic traits of RTX toxin proteins. The S-layer and RTX domains of Csx are separated by a proline-rich stretch of 48 amino acids. All C. rectus isolates studied contained copies of either the csxA or csxB gene or both; csx genes were absent from all other Campylobacter and Helicobacter species examined. Serum of a patient with acute gingivitis showed a strong reaction to recombinant Csx protein on immunoblots.

Controversies of fetal cardiac intervention

Remarkable advances in ultrasound imaging technology have made it possible to diagnose fetal cardiovascular lesions as early as 12-14 weeks of gestation and to assess their physiological relevance by echocardiography. Moreover, invasive techniques have been developed and refined to relieve significant congenital heart disease (CHD), such as critical aortic and pulmonary stenoses in the pediatric population including neonates. Recognition of the fact that certain CHDs can evolve in utero, and early intervention may improve the outcome by altering the natural history of such conditions has led to the evolution of a new fetal therapy, i.e. fetal cardiac intervention. Two entities, pulmonary valvar atresia and intact ventricular septum (PA/IVS) and hypoplastic left heart syndrome (HLHS), are associated with significant morbidity and mortality even with postnatal surgical therapy. These cases are believed to occur due to restricted blood flow, leading to impaired growth and function of the right or left ventricle. Therefore, several centers started the approach of antenatal intervention with the primary goal of improving the blood flow through the stenotic/atretic valve orifices to allow growth of cardiac structures. Even though centers with a reasonable number of cases seem to have improved the technique and the immediate outcome of fetal interventions, the field is challenged by ethical issues as the intervention puts both the mother and the fetus at risk. Moreover, the perceived benefits of prenatal treatment have to be weighed against steadily improving postnatal surgical and hybrid procedures, which have been shown to reduce morbidity and mortality for these complex heart defects. This review is an attempt to provide a balanced opinion and an update on fetal cardiac intervention.

A1.40 Adiponektin and resistin levels in pregnant patients with rheumatoid arthritis

BACKGROUND Pregnancy induces a modulation of the maternal immune system in order to install tolerance towards the semiallogeneic fetus. This change of the maternal immune systems influences some autoimmune diseases such as rheumatoid arthritis (RA) in a positive way. Our previous study showed that genes of the adipocytokine pathway were differently regulated by pregnancy as well as by RA. The objective of this study was to analyse the association between pregnancy induced improvement of RA and changes of adipocytokine levels. MATERIAL AND METHODS Adiponectin and resistin levels were measured in sera of pregnant (n = 29) and non-pregnant (n = 24) RA patients as well as in pregnant (n = 26) and non-pregnant (n = 9) healthy controls by ELISA. Pregnant RA patients were analysed before conception, once at each trimester and 8 weeks postpartum. Disease activity was measured by CRP and DAS28-CRP. RESULTS Resistin levels were higher in non-pregnant RA patients than in healthy controls. Resistin levels increased during pregnancy and decreased postpartum in both healthy subjects and RA patients. However, RA patients with active disease during pregnancy showed higher resistin levels at the third trimester than healthy women. There was a positive correlation between resistin levels and CRP. Adiponektin levels increased at the second trimester of pregnancy and decreased thereafter in both healthy subject and RA patients. There was no difference between patients and healthy subjects. Adiponektin levels of RA patients negatively correlated with CRP. CONCLUSION Pregnancy induces an increase of both the resistin and the adiponectin levels. Resistin levels are further influenced by active disease. By contrast, the increase of the adiponectin levels at the second trimester might play a role in the modulation of disease activity of RA.

The small SLC43 family: facilitator system l amino acid transporters and the orphan EEG1

The SLC43 family is composed of only three genes coding for the plasma membrane facilitator system l amino acid transporters LAT3 (SLC43A1; TC 2.A.1.44.1) and LAT4 (SLC43A2; TC 2.A.1.44.2), and the orphan protein EEG1 (SLC43A3; TC 2.A.1.44.3). Besides the known mechanism of transport of LAT3 and LAT4, their physiological roles still remain quite obscure. Morphants suggested a role of LAT3 in renal podocyte development in zebrafish. Expression in liver and skeletal muscle, and up-regulation by starvation suggest a role of LAT3 in the flux of branched-chain amino acids (BCAAs) from liver and skeletal muscle to the bloodstream. Finally, LAT3 is up-regulated in androgen-dependent cancers, suggesting a role in mTORC1 signaling in this type of tumors. In addition, LAT4 might contribute to the transfer of BCAAs from mother to fetus. Unfortunately, the EEG1 mouse model (EEG1(Y221∗)) described here has not yet offered a clue to the physiological role of this orphan protein.

Use of a Th1 Stimulator Adjuvant for Vaccination against Neospora caninum Infection in the Pregnant Mouse Model

Vertical transmission from an infected cow to its fetus accounts for the vast majority of new Neospora caninum infections in cattle. A vaccine composed of a chimeric antigen named recNcMIC3-1-R, based on predicted immunogenic domains of the two microneme proteins NcMIC1 and NcMIC3, the rhoptry protein NcROP2, and emulsified in saponin adjuvants, significantly reduced the cerebral infection in non-pregnant BALB/c mice. Protection was associated with a mixed Th1/Th2-type cytokine response. However, the same vaccine formulation elicited a Th2-type immune response in pregnant mice and did not prevent vertical transmission or disease, neither in dams nor in offspring mice. In this study, an alternative vaccine formulation containing recNcMIC3-1-R emulsified in Freund’s incomplete adjuvant, a stimulator of the cellular immunity, was investigated. No protection against vertical transmission and cerebral infection in the pregnant mice and a very limited protective effect in the non-pregnant mice were observed. The vaccine induced a Th1-type immune response characterized by high IgG2a titres and strong IFN-γ expression, which appeared detrimental to pregnancy.

Stellenwert von Gerinnungsstörungen bei habituellen Aborten

Habituelle Aborte Ein Spontanabort ereignet sich bei etwa 15 % aller klinisch festgestellten Schwangerschaften. Vom betroffenen Paar wird er ausnahmslos als äußerst traumatisch erlebt. Insbesondere gilt dies beim habituellen Abort (≥ 3 Aborte in Folge), der etwa 1 % der Schwangerschaften betrifft. In der Hoffnung, weitere Aborte zu verhindern, werden entsprechend große Anstrengungen unternommen, die jeweilige Ursache zu eruieren. Gerinnungsphysiologische Einflüsse Pathophysiologisch spielen nebst organischen und zytogenetischen Anomalien beim Fetus vermutlich auch gerinnungsphysiologische Einflüsse eine ursächliche Rolle, insbesondere erworbene und hereditäre prokoagulatorische Störungen. Diese können das im Rahmen der Schwangerschaft schon physiologisch erhöhte Gerinnungspotenzial zusätzlich verstärken und damit die Blutversorgung des Fetus potenziell behindern, was mit der Gefahr seiner Abstoßung einhergeht. Thrombophilie Auch wenn der diesbezügliche Beweis im Einzelfall schwierig zu erbringen ist, erscheint eine ungünstige Beeinflussung des Abortrisikos durch erworbene und hereditäre Thrombophilien plausibel. Daraus ergibt sich unschwer die Folgerung oder Hoffnung, dass antiaggregatorische und antikoagulatorische Maßnahmen eine günstige Wirkung haben könnten. Der vorliegende Beitrag geht auf die bekannten sowie teils auch nur vermuteten pathophysiologischen Mechanismen und die sich daraus ergebenden therapeutischen bzw. präventiven Möglichkeiten ein.

The long non-coding RNA NEAT1 is increased in IUGR placentas, leading to potential new hypotheses of IUGR origin/development.

INTRODUCTION Intrauterine Growth Restriction (IUGR) is a multifactorial disease defined by an inability of the fetus to reach its growth potential. IUGR not only increases the risk of neonatal mortality/morbidity, but also the risk of metabolic syndrome during adulthood. Certain placental proteins have been shown to be implicated in IUGR development, such as proteins from the GH/IGF axis and angiogenesis/apoptosis processes. METHODS Twelve patients with term IUGR pregnancy (birth weight < 10th percentile) and 12 CTRLs were included. mRNA was extracted from the fetal part of the placenta and submitted to a subtraction method (Clontech PCR-Select cDNA Subtraction). RESULTS One candidate gene identified was the long non-coding RNA NEAT1 (nuclear paraspeckle assembly transcript 1). NEAT1 is the core component of a subnuclear structure called paraspeckle. This structure is responsible for the retention of hyperedited mRNAs in the nucleus. Overall, NEAT1 mRNA expression was 4.14 (±1.16)-fold increased in IUGR vs. CTRL placentas (P = 0.009). NEAT1 was exclusively localized in the nuclei of the villous trophoblasts and was expressed in more nuclei and with greater intensity in IUGR placentas than in CTRLs. PSPC1, one of the three main proteins of the paraspeckle, co-localized with NEAT1 in the villous trophoblasts. The expression of NEAT1_2 mRNA, the long isoform of NEAT1, was only modestly increased in IUGR vs. CTRL placentas. DISCUSSION/CONCLUSION The increase in NEAT1 and its co-localization with PSPC1 suggests an increase in paraspeckles in IUGR villous trophoblasts. This could lead to an increased retention of important mRNAs in villous trophoblasts nuclei. Given that the villous trophoblasts are crucial for the barrier function of the placenta, this could in part explain placental dysfunction in idiopathic IUGR fetuses.

Characterization of a novel CYP19A1 (aromatase) R192H mutation causing virilization of a 46,XX newborn, undervirilization of the 46,XY brother, but no virilization of the mother during pregnancies.

BACKGROUND P450 aromatase (CYP19A1) is essential for the biosynthesis of estrogens from androgen precursors. Mutations in the coding region of CYP19A1 lead to autosomal recessive aromatase deficiency. To date over 20 subjects have been reported with aromatase deficiency which may manifest during fetal life with maternal virilization and virilization of the external genitalia of a female fetus due to low aromatase activity in the steroid metabolizing fetal-placental unit and thus high androgen levels. During infancy, girls often have ovarian cysts and thereafter fail to enter puberty showing signs of variable degree of androgen excess. Moreover, impact on growth, skeletal maturation and other metabolic parameters is seen in both sexes. OBJECTIVE AND HYPOTHESIS We found a novel homozygous CYP19A1 mutation in a 46,XX girl who was born at term to consanguineous parents. Although the mother did not virilize during pregnancy, the baby was found to have a complex genital anomaly at birth (enlarged genital tubercle, fusion of labioscrotal folds) with elevated androgens at birth, normalizing thereafter. Presence of 46,XX karyotype and female internal genital organs (uterus, vagina) together with biochemical findings and follow-up showing regression of clitoral hypertrophy, as well as elevated FSH suggested aromatase deficiency. Interestingly, her older brother presented with mild hypospadias and bilateral cryptorchidism and was found to carry the same homozygous CYP19A1 mutation. To confirm the clinical diagnosis, genetic, functional and computational studies were performed. METHODS AND RESULTS Genetic analysis revealed a homozygous R192H mutation in the CYP19A1 gene. This novel mutation was characterized for its enzymatic activity (Km, Vmax) in a cell model and found to have markedly reduced catalytic activity when compared to wild-type aromatase; thus explaining the phenotype. Computational studies suggest that R192H disrupts the substrate access channel in CYP19A1 that may affect binding of substrates and exit of catalytic products. CONCLUSION R192H is a novel CYP19A1 mutation which causes a severe phenotype of aromatase deficiency in a 46,XX newborn and maybe hypospadias and cryptorchidism in a 46,XY, but no maternal androgen excess during pregnancy.

Influence of transrectal and transabdominal ultrasound examination on salivary cortisol, heart rate, and heart rate variability in mares

Pregnancy diagnostics in equine reproduction are routinely performed using transrectal ultrasonography, although it is also possible to visualize the fetus by transabdominal ultrasound examinations from the 90th day of gestation onward. We hypothesized that ultrasound examinations may stress the mare and that the gestational stage status and lactation may influence the mare's stress reaction. To investigate the stress reaction, 25 thoroughbred mares of different age, pregnancy and lactational status underwent a transrectal examination. In pregnant mares, an additional transabdominal examination was performed. Salivary cortisol concentration, mean heart rate, and heart rate variability of mares were assessed to evaluate the reactions of hypothalamic–pituitary–adrenal (HPA) axis and of the autonomic nervous system. Significant differences were observed between lactating and nonlactating mares; with a lower responsiveness to stress in lactating mares. The transrectal ultrasound examination in nonlactating mares induced a significant increase in salivary cortisol (P < 0.05), and in the heart rate variability parameter, ratio of low to high frequencies (P < 0.05). This reflects an activation of the HPA axis and a shift to more sympathetic dominance. In contrast, a transabdominally performed pregnancy check did not induce an activation of the HPA axis over basal level but increased the mean heart rate and low to high frequency ratio. The results of this study indicate that checks of advanced pregnancies can be easily performed by transabdominal ultrasonography. With regard to animal welfare, this technique should be preferred during midgestation in nonlactating mares.

Laboratory diagnosis of ruminant abortion in Europe.

Abortion in ruminants is a major cause of economic loss worldwide, and the management and control of outbreaks is important in limiting their spread, and in preventing zoonotic infections. Given that rapid and accurate laboratory diagnosis is central to controlling abortion outbreaks, the submission of tissue samples to laboratories offering the most appropriate tests is essential. Direct antigen and/or DNA detection methods are the currently preferred methods of reaching an aetiological diagnosis, and ideally these results are confirmed by the demonstration of corresponding macroscopic and/or histopathological lesions in the fetus and/or the placenta. However, the costs of laboratory examinations may be considerable and, even under optimal conditions, the percentage of aetiological diagnoses reached can be relatively low. This review focuses on the most commonly occurring and important abortifacient pathogens of ruminant species in Europe highlighting their epizootic and zoonotic potential. The performance characteristics of the various diagnostic methods used, including their specific advantages and limitations, are discussed.

PLACENTAL GLUCOSE TRANSPORTER (GLUT)-1 REGULATION IN PREECLAMPSIA

PLACENTAL GLUCOSE TRANSPORTER (GLUT)-1 REGULATION IN PREECLAMPSIA Camilla Marini a,b, Benjamin P. Lüscher a,b, Marianne J€orger-Messerli a,b, Ruth Sager a,b, Xiao Huang c, Jürg Gertsch c, Matthias A. Hediger c, Christiane Albrecht c, Marc U. Baumann a,c, Daniel V. Surbek a,c a Department of Obstetrics and Gynecology, University Hospital of Bern, Bern, Switzerland, Switzerland; b Department of Clinical Research, University of Bern, Bern, Switzerland, Switzerland; c Institute for Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland, Switzerland Objectives: Glucose is a primary energy source for the fetus. The absence of significant gluconeogenesis in the fetus means that the fetal up-take of this vital nutrient is dependent on maternal supply and subsequent transplacental transport. Altered expression and/or function of placental transporters may affect the intrauterine environment and could compromise fetal and mother well-being. We speculated that pre-eclampsia (PE) impairs the placental glucose transport system. Methods: Placentae were obtained after elective caesarean sections following normal pregnancies and pre-eclamptic pregnancies. Syncytial basal membrane (BM) and apical microvillus membrane (MVM) fractions were prepared using differential ultra-centrifugation and magnesium precipitation. Protein expression was assessed by western blot analysis. mRNA levels in whole villous tissue lysate were quantified by real-time PCR. To assess glucose transport activity a radiolabeled substrate up-take assay and a transepithelial transport model using primary cytotrophoblasts were established. Results: GLUT1 mRNA expression was not changed in PE when compared to control, whereas protein expression was significantly down-regulated. Glucose up-take into syncytial microvesicles was reduced in PE compared to control. In a transepithelial transport model, phloretinmediated inhibition of GLUT1 at the apical side of primary cytotrophoblasts showed a 44% of reduction of transepithelial glucose transport at IC50. Conclusions: GLUT1 is down-regulated on protein and functional level in PE compared to control. Altering glucose transport activity by inhibition of apical GLUT-1 indicates that transplacental glucose transport might be regulated on the apical side of the syncytiotrophoblast. These results might help to understand better the regulation of GLUT1 transporter and maybe in future to develop preventive strategies to modulate the fetal programming and thereby reduce the incidence of disease for both the mother and her child later in life.

Placental Uric Acid Transport System: Glucose Transporter 9 (SLC2A9)

Placental Uric Acid Transport System: Glucose Transporter 9 (SLC2A9). INTRODUCTION: Pre-eclampsia, a pregnancy-specific disease, contributes substantially to perinatal morbidity and mortality of both the mother and her child. Pre-eclampsia is often associated with high maternal urate serum levels, which in turn has been shown to play a role in the pathogenesis of this disease. The aim of this study was to investigate the glucose transporter GLUT9-mediated placental uric acid transport system. METHODS: In this study western blot, immunofluorescence techniques as well as a transepithelial transport (Transwell) model were used to assess GLUT9 protein expression and, respectively, uric acid transport activity. Electrophysiological techniques and transmission electron microscopy (TEM) were used to characterize the properties and the structure of GLUT9. RESULTS: Uric acid is transported across a BeWo choriocarcinoma cell monolayer with 530 pmol/min. We could successfully overexpress and for the first time purify the GLUT9b isoform using the Xenopus laevis oocytes expression system. Chloride seems to modulate the urate transport system. TEM revealed that GLUT9b isoform is present as monomer and dimmer in the Xenopus laevis overexpression model. A class average of all the particles allowed us to develop a first model of human GLUT9b structure, which was derived from the published crystal structure of the bacterial homologue of GLUT1-4. CONCLUSIONS: In vitro the “materno-fetal” transport of uric acid is slow indicating that in vivo the fetus might be protected from short-term fluctuations of maternal urate serum levels. The low-resolution structure obtained from TEM validates the proposed homology model regarding the structure of human GLUT9b. In ongoing studies this model is used to perform virtual screening to identify novel modulators of the urate transport system enabling the development of novel therapies in pregnancy complications.