Physcomitrella patens Reute mCherry as a tool for efficient crossing within and between ecotypes.

Physcomitrella patens is a monoecious moss that is predominantly selfing in the wild. Laboratory crossing techniques have been established and crosses between the sequenced Gransden ecotype and the genetically divergent Villersexel ecotype were used for genetic mapping. The recently introduced ecotype Reute has a high fertility rate and is genetically more closely related to the Gransden ecotype than the Villersexel ecotype. Reute sexual reproduction phenology is similar to Gransden, which should allow successful crossing. Using the Reute ecotype and an existing Gransden mutant as a test case, we applied a normalised crossing approach to demonstrate crossing potential between these ecotypes. Also, using a standard transformation approach, we generated Reute fluorescent strains expressing mCherry that allow an easy detection of crossed offspring (sporophyte). We show that Reute can be successfully crossed with a self-infertile DR5:DsRed2 mutant generated in the Gransden background. Using newly established Reute fluorescent strains, we show that they can efficiently fertilise Reute as well as Gransden wild type. The resulting progeny display Mendelian 1:1 segregation of the fluorescent marker(s), demonstrating the suitability of such strains for genetic crossing. Overall our results demonstrate that Reute is highly suitable for genetic crossing. The Reute mCherry strain can be used as a suitable background for offspring selection after crossing.

The clinical significance of antenatal pathological Doppler findings in the fetal middle cerebral artery in cases with peripheral reduced diastolic doppler flow but no absence of end-diastolic flow in the umbilical artery or fetal aorta.

PURPOSE: The effects of antepartum pathological Doppler findings in the fetal middle cerebral artery in cases with simultaneously peripheral reduced diastolic Doppler flow on perinatal outcome and the odds ratio of perinatal risks were studied. METHODS: 214 patients were examined by color Doppler ultrasound in relationship to complications in gestation and labor and fetal outcome. One thousand and seventy Doppler flow measurements of the middle cerebral artery, the umbilical artery and the fetal aorta between 28 and 40 weeks of gestation were performed. Sensitivity and odds ratio of synchronous cerebral pathological and peripheral pathological Doppler blood flow with regard to the prediction of intrauterine growth retardation, rate of cesarean section, preterm delivery and newborn depression was calculated. In all Doppler measurements there were no cases with absence of end-diastolic flow. RESULTS: Preterm delivery rate and intrauterine growth retardation rate were significantly higher in cases of synchronous cerebral pathological and peripheral pathological Doppler blood flow as in cases of isolated reduced peripheral blood flow (p<0.001; odds ratio 13.2 and 16.6). CONCLUSION: Pregnancies with no absence of end-diastolic flow in the fetal aorta or umbilical artery, but with reduced diastolic flow in these vessels and simultaneous pathological Doppler findings in the fetal middle cerebral artery are high risk pregnancies, above all in respect to intrauterine growth retardation, preterm delivery and newborn depression. Surveillance of pregnant women should be performed in a perinatal centre.

The clinical significance of antenatal pathological Doppler findings in fetal middle cerebral artery compared to umbilical artery and fetal aorta.

PURPOSE: To compare the diagnostic capacity of fetal Doppler velocimetry in fetal middle cerebral artery to umbilical artery and fetal aorta to the prediction of fetal outcome. METHODS: 229 patients between 28 weeks of gestation (weeks) and 40 weeks were examined by Doppler ultrasound in relationship to complications in gestation and labor, and fetal outcome. One thousand two hundred and seventy doppler blood flow velocity waveforms in the middle cerebral artery, umbilical artery and fetal aorta were recorded. Sensitivity of these vessels with regard to the prediction of intrauterine growth retardation, rate of cesarean section, preterm delivery and new-born depression was calculated. In all Doppler measurements there were no cases with absence of end-diastolic flow. RESULTS: The differences between pathological fetal outcome were not statistically significant, but diagnostic capacity of the middle cerebral artery velocimetry with regard to the prediction of pathological fetal outcome was on average 11% below the diagnostic capacity of the fetal aorta and umbilical artery. CONCLUSION: There is no benefit in examining fetal middle cerebral artery Doppler measurements in clinical routine in patients with normal velocity waveforms in the umbilical artery and fetal aorta.

Reference ranges and standard percentile-curves for the Doppler indices RI and S/D ratio of the fetal middle cerebral artery. Color Doppler measurements in a perinatal centre.

PURPOSE: To perform reference ranges and standard percentile-curves for the Doppler indices resistance index (RI) and systolic/diastolic (S/D) ratio of the fetal middle cerebral artery, periodical color doppler sonographic measurements of 70 pregnant women in the 3rd trimester of pregnancy were done. METHODS: 600 Doppler flow measurements of the fetal middle cerebral artery between 28 and 40 weeks of gestation were performed. The patients had no previous obstetric complications, no apparent medical problems and no complications in pregnancy and labor. Percentiles curves were performed for the RI and the S/D ratio from the 10th percentile up to the 90th percentile. RESULTS: In the 3rd trimester of pregnancy fetal cerebral circulation shows an increase of the diastolic component and simultaneous decrease in cerebral resistance. The average S/D ratio in week 29 is 8.0, in week 34 6.0 and in the 40th week 3.5. The RI decrement is from 0.88 to 0.67. Other groups demonstrated similar ranges however absolute data are different. CONCLUSION: The increase in the diastolic component in the middle cerebral artery of the last third of the pregnancy demands reference ranges by using percentile curves. Knowledge of the reference range helps to discriminate between a normal fetal situation and disease. Because of different absolute ranges in the literature each perinatal centre should develop their own data.

Evaluation of the secretion of the atrial natriuretic factor (ANF) after laparotomy.

BACKGROUND: ANF is a potent diuretic, natriuretic and vasorelaxant hormone. The objective of the present study was to examine the effect of opioid receptor stimulation by morphine after surgery on endogenous ANF production and diuresis. METHODS: Prospectively, 11 women undergoing surgery for either uterine leiomyomas, chronic pelvic discomfort or desire for definitive contraception by laparotomy were evaluated. Venous samples were collected at fixed times. Concentrations of ANF were measured by commercially available radioimmunoassay test kits. Statistical analysis was performed by the Friedman Two way ANOVA. Kruskal-Wallis 1-way ANOVA and Mann-Whitney U-Wilcoxon Rank Sum W Test. The level of significance was set at probability below 0.05. RESULTS: There were statistically significant changes in the serum levels of ANF (p=0.0028), in pain score (p<0.0001) and urinary flow rate (p<0.0001) after operation, while the diastolic (p=0.0671) and systolic (p=0.0543) blood pressure showed slightly significant changes. CONCLUSION: Our results show that i.v. administered morphine induces a potent diuretic effect via activation of opioid receptors and suggest that this effect is due to the enhanced release of ANF. However the mechanism by which morphine induces the ANF release remains to be evaluated.

Heterologous mullerian mixed tumor after whole body irradiation because of Hodgkin's disease in stage IV.

PURPOSE: With an incidence of 1.5% of all malignant diseases of the uterus as specified in the literature, the mullerian mixed tumor is a rarity amongst the malignancies of the female genital tract. METHODS: A retrospective analysis of an individual case report with the occurrence of heterologous mullerian mixed tumor years after irradiation because of Hodgkin's disease. RESULTS: This case reports describes the occurrence of a mullerian mixed tumor 12 years after the treatment of Hodgkin's disease by whole body irradiation. To our knowledge, the incidence of a mullerian mixed tumor after the treatment of Hodgkin's disease has rarely been described up to now in the literature. CONCLUSION: This case report appears to indicate the possible carcinogenic potency of radiotherapy when administered many years before. A causal connection between the administration of whole body irradiation and the development of a mullerian mixed tumor cannot be established.

Atrial natriuretic factor (ANF) after laparoscopy and morphine application for pain therapy.

OBJECTIVE: ANF is a potent diuretic, natriuretic and vasorelaxant hormone. The objective of the present study was to examine the effect of opioid receptor stimulation by morphine on endogenous ANF production and diuresis after surgery. METHODS: Prospectively, nine women undergoing surgery for either uterine leiomyomas, chronic pelvic discomfort or desire for definitive contraception by laparoscopy were evaluated. Venous samples were collected at fixed times. Concentrations of ANF were measured by commercially available radioimmunassay test kits. Statistical analysis was performed by the Friedman Two-way ANOVA, Kruskal-Wallis 1-way ANOVA and Mann-Whitney U-Wilcoxon Rank Sum W Test. The level of significance was set at a probability below 0.05. RESULTS: There were no statistically significant changes in serum levels of ANF (p = 0.98), in diastolic blood pressure (p = 0.14) or pain score (p = 0.86) after surgery. Systolic blood pressure (p = 0.0032), pulse rate (p = 0.019) and urinary flow rate (p < 0.0001) showed significant changes during observation. CONCLUSION: Our results show that i.v. administered morphine induces a potent diuretic effect via activation of opioid receptors but it can not be suggested that this effect is due to an enhanced release of ANF.

Postpartum ovarian vein thrombosis.

OBJECTIVE: Ovarian vein thrombosis (OVT) is known as a rare but serious postpartum complication. The condition is often clinically not distinguishable from endometritis, appendicitis or pyelonephritis. OVT may cause sepsis, septic pulmonary thromboembolism, and thrombosis of the inferior vena cava and the renal veins, and is potentially fatal. The objective of this study was to report the clinical findings and outcome of two patients with diagnosed ovarian vein thrombosis after delivery managed at this institution. METHOD: Two patients fit the study criteria of documented ovarian vein thrombosis after delivery. An imaging diagnosis (CT) of ovarian vein thrombosis was required for final study inclusion. RESULTS: We present two patients with ovarian vein thrombosis. The symptoms of one patient disappeared two days after beginning heparin and antibiotic therapy. The control-CT 93 days after the diagnosis of POVT showed unsuspected ovarian veins. The other patient suffered from POVT 13 days after spontaneous delivery. Because of lethal embolisms she died during the operation for embolectomy. CONCLUSION: On the basis of our series and other recent series, OVT may likely be more common than previously thought and may become clinically apparent only when complicated by infection, expansion of the thrombus or pulmonary embolism. POVT is a potentially fatal condition most commonly seen as a complication of pelvic surgery or inflammatory disease.

[Intrauterine growth retardation--perinatal mortality and postnatal morbidity in a perinatal center]. / Die intrauterine Wachstumsretardierung--Perinatale Mortalität und postnatale Morbidität an einem Perinatalzentrum.

OBJECTIVES: The intrauterine growth retardation (IUGR) contributes specially to perinatal mortality and morbidity. An effective treatment is not yet available. The purpose of this study is to analyse the perinatal data of a cohort with IUGR-infants including mortality and morbidity and to describe prematurity and IUGR as a combination of high risk. STUDY DESIGN: We have analysed the perinatal data of a cohort of 220 patients with IUGR-infants including mortality and morbidity. Regularly we examined fetal ultrasonographic growth, doppler measurements of fetal and maternal vessels and CTG, as well as indicated amniocentesis, placentesis and cordocentesis. Entry criteria were: normal menstrual period before pregnancy, clear gestational age, complete history of pregnancy. RESULTS: At the Department of Gynaecology & Obstetrics, University Hospital, Homburg/Saar, the incidence of IUGR (< 10th Percentile) in premature babies and newborns was 13.1% over five years (1993-1997). The percentage of premature babies with IUGR was 18.6%. The overall mortality was 4.1%, the perinatal mortality was 3.6% and the neonatal mortality was 2.7%. In 1.4% intrauterine foetal death was observed, all associated with a birthweight below the 3th percentile. The average duration of pregnancy was 34 + 4 weeks. A high perinatal mortality of 13.1% and an overall mortality of 14.7% was observed. 11.4% of the premature babies with IUGR showed deformities and about 2% presented chromosome aberrations. 46.4% of children have been transferred to the Neonatal Intensive Care Unit. 57.1% of the postnatal complications have been related to the lungs, 26.5% to the cardiovascular system and 14.3% to the cerebrum. Maternal complications were referred to: SIH/EPH-gestosis (65.4%), HELLP-syndrome (5.8%), nicotine abuse (31.4%), pregnancy anaemia (17.3%) and gestational diabetes with insuline therapy (7.7%). On the placental site the most noticeable conditions have been placental insufficiency (40.6%) and placental infarction (28.7%). CONCLUSION: Children with IUGR are exposed to high perinatal mortality and postnatal morbidity. Premature babies in association with an IUGR are at high risk. The surveillance of the pregnant women and the new-born children should be performed in a perinatal centre.

Immunohistochemical detection of apoptosis in the human term and post-term placenta.

PURPOSE: To study the incidence of apoptosis in human term and post-term placenta and to determine its presence in different areas of placentas of uncomplicated pregnancies. METHODS: A total of 15 placentas, 8 obtained from spontaneous deliveries and elective caesarean sections at term (37-41 weeks of pregnancy) and 7 from spontaneous deliveries and elective caesarean sections post-term (> 41 weeks of pregnancy) were included in this study. Apoptosis was identified by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling method (TUNEL, Boehringer, Mannheim, Germany) in paraffin-embedded sections. RESULTS: Apoptosis was predominantly detected in trophoblast and stromal tissue. There were no significant differences in the incidence of apoptosis in different parts of placental tissue. A significant increase of apoptosis was seen in both trophoblast and stromal cells of post-term placentas (p < 0.05; p < 0.005). CONCLUSIONS: Apoptosis could be detected in the human term and post-term placenta, with increasing incidence in post-term placental tissue, suggesting a possible role of apoptosis in the mechanism of parturition and placental senescence.

Immunohistochemical analysis of DNA mismatch-repair enzyme hMSH-2 and Ki-67 in breast carcinomas.

BACKGROUND: The human Mut-S-Homologon-2 (hMSH-2) gene product is a member of a highly conserved family of proteins involved in postrepiclation mismatch repair. MATERIALS AND METHODS: We have analyzed hMSH-2 expression in normal breast tissue (n = 10) and breast carcinomas (n = 30). hMSH-2 protein was investigated immunohistochemically on frozen sections using a specific mouse monoclonal antibody (clone FE11). hMSH-2 labelling pattern was compared with the staining pattern of the proliferation marker Ki-67. A hMSH-2-immunoreactivity score (hMSH-2-IRS) for the semiquantitative analysis of hMSH-2 expression is presented. RESULTS: In normal breast tissue, we only found weak nuclear immunoreactivity for hMSH-2 in 70%, while the remaining 30% were hMSH-2 negative (mean hMSH-2- IRS: 1.00; SD: +/- 0.82). All breast carcinomas analyzed revealed moderate to strong nuclear immunoreactivity (mean hMSH-2-IRS: 7.67; SD: +/- 3.55). hMSH-2 staining was heterogeneous, with visual differences between individual tumour cells. Expression of hMSH-2 protein was consistently and strongly upregulated in tumour cells of breast carcinomas as compared to normal breast tissue. No visual correlation in comparing the labelling patterns for hMSH-2 with the labeliing patterns for Ki-67 (mean percentage of Ki-67 positive tumour cells: 24.33%; SD: +/- 15.35) was observed in breast carcinomas. CONCLUSION: Our findings indicate that (a) hMSH-2 is expressed in normal human breast tissue; (b) expression of hMSH-2 may be of importance for the genetic stability of breast carcinomas in vivo.

Immunohistochemical evaluation of apoptosis in placentae from normal and intrauterine growth-restricted pregnancies.

OBJECTIVE: To investigate the extent of apoptosis within the human placenta in tissues from normotensive term pregnancies and those complicated by intrauterine growth-restriction (IUGR). METHODS: A total of 18 placentas, 10 obtained from uncomplicated term gestations and 8 from intrauterine growth restricted fetuses were included in this study. Apoptosis was identified using a terminal deoxynucleotidyl transferase-mediated deoxyuridine triophosphate nick end-labeling technique (TUNEL, Boehringer, Mannheim, Germany) in paraffin-embedded sections. RESULTS: Apoptosis was predominantly detected in the villous trophoblast and stromal tissue. There were no differences in the incidence of apoptosis in different parts of the same placenta. The apoptotic index in placental tissue from uncomplicated pregnancies was 0.93 +/- 0.12. Significantly more apoptotic nuclei were detected in the placental tissue from IUGR gestation (4.2 +/- 2.96, p < 0.01). CONCLUSION: These results might point toward a possible role of apoptosis in the pathophysiology of intrauterine growth-restriction.

Immunohistochemical analysis of DNA 'mismatch-repair' enzyme human Mut-S-Homologon-2 in ovarian carcinomas.

The human Mut-S-Homologon-2 (hMSH-2) gene product is a member of a highly conserved family of proteins involved in postreplication mismatch repair. We have analysed hMSH-2 expression in normal ovarian tissue (n = 15) and ovarian carcinomas (n = 40). hMSH-2 protein was investigated immunohistochemically on frozen sections using a highly sensitive streptavidin-peroxidase technique and a specific mouse monoclonal antibody (clone FE11). A hMSH-2-immunoreactivity score (hMSH-2-IRS) for semiquantitative analysis of hMSH-2 expression is presented. In normal ovarian tissue, we only found weak nuclear immunoreactivity for hMSH-2 in 60%, while the remaining 40% were hMSH-2 negative (mean hMSH-2-IRS: 0.73; SD: +/-0.70). All ovarian carcinomas analysed revealed moderate to strong nuclear immunoreactivity (mean hMSH-2-IRS: 8.05; SD: +/-3.65). hMSH-2 staining was heterogeneous, with visual differences between individual tumour cells. Expression of hMSH-2 protein was consistently and strongly upregulated in tumour cells of ovarian carcinomas as compared to normal ovarian tissue. No statistically significant correlation in comparing the labelling patterns for hMSH-2 with the labelling patterns for Ki-67 (mean percentage of Ki-67 positive tumour cells: 25.88%; SD: +/-18.43) was observed in ovarian carcinomas. Furthermore, no statistical significant correlations between hMSH-2-IRS and histological grading (p = 0.47), histological type of carcinoma (p = 0.706) or FIGO-classification (p = 0.054) were found. Our findings indicate that (a) hMSH-2 is expressed in normal human ovarian tissue, (b) expression of hMSH-2 is increased in ovarian carcinomas, (c) expression of hMSH-2 may be of importance for the genetic stability of ovarian carcinomas in vivo, (d) hMSH-2 mutations may not cause microsatellite instability in ovarian carcinomas, (e) hMSH-2 may contribute to mechanisms responsible for resistance to anticancer drugs.