Mild ventriculomegaly, mild cerebellar hypoplasia and dysplastic choroid plexus as early prenatal signs of CHARGE association.

CHARGE association has been diagnosed postnatally in increasing numbers since the first description in 1979. The acronym CHARGE includes the abnormalities ocular Colobomas (iris, retina or nervus opticus), Heart disease, Atresia of choanae, Retarded growth and development and/or central nervous system anomalies, Genital hypoplasia, Ear anomalies and deafness. So far, no prenatal diagnosis of the CHARGE association has been described; only one case report presents prenatal symptoms detected at 31 gestational weeks. In our case, prenatally detected mild cerebral ventriculomegaly and dysplasia of choroid plexus were abnormalities visible as early as 15+6 weeks as well as mild cerebellar hypoplasia at 21+1 weeks. At 28+6 weeks, in addition polyhydramnios could be found. The combination of the 'benign' central-nervous findings raised suspicion of a severe congenital malformation at 21+1 weeks which was confirmed postnatally in the form of diagnosis of CHARGE association.

Prenatal diagnosis of familial absent pulmonary valve syndrome: case report and review of the literature.

We report on a case of absent pulmonary valve syndrome in a woman with a history of one healthy child and one child with tetralogy of Fallot with absent pulmonary valve. The diagnosis was missed at the first ultrasound examination performed at 13 + 5 weeks of gestation and correctly diagnosed at 21 + 5 weeks. Re-evaluation of the ultrasound examination recorded at 13 + 5 weeks exhibited severe insufficiency of the pulmonary valve at this time. However, neither dilatation of the right and left pulmonary arteries nor asymmetry of the ventricles were present at that time. The pregnancy was terminated at 22 + 1 weeks of gestation when autopsy confirmed the diagnosis of absent pulmonary valve syndrome. Karyotyping of the fetus after termination of pregnancy revealed normal chromosomes. Echocardiography of the parents and the healthy sibling revealed normal results.

Sonographic markers of exencephaly at 9 + 3 weeks of gestation.

Following the introduction of transvaginal sonography, several first trimester diagnoses of the exencephaly anencephaly sequence have been reported, with the earliest being described at 10 weeks of gestation. We report a case with a high index of suspicion for exencephaly at 9 weeks and 3 days because of three sonographic features: the cranial pole of the embryo was smaller than the chest, the cranial pole bulged dorsally and the surface of the cranium was irregular. The diagnosis was confirmed by a repeat ultrasound examination at 11 + 4 weeks and by autopsy after termination of pregnancy.

Histochemical evaluation of placental dipeptidyl peptidase IV (CD26) in pre-eclampsia: enzyme activity in villous trophoblast indicates an enhanced likelihood of gestational hypertensive disorders.

PROBLEM: To determine whether differences in placental dipeptidyl peptidase IV (DPP IV, CD26) activities occurred in hypertensive complicated pregnancies as compared with uncomplicated pregnancies. METHOD: DPP IV activity was detected with H-Gly-Pro-4M2NA as the substrate in placental cryostat sections from 65 patients with gestational hypertension and 67 patients with uncomplicated pregnancies. The graduated intensities of the reaction product in the villous trophoblast were scored semiquantitatively by light microscopy and were related to the relative frequencies of hypertensive disorders (proportional odds model). After detection of enzyme activity, the same tissue samples were homogenized and used for kinetic fluorometric measurements. RESULTS: Enhanced villous trophoblastic DPP IV activity was significantly associated with an increased frequency of proteinuric hypertension in pregnant women (cumulative odds ratio theta1 = 1.6; P < 0.01). CONCLUSION: This study demonstrates for the first time that increased villous trophoblastic DPP IV activity indicates an increased likelihood of the presence and of the severity of hypertensive disorders in pregnancy.

Binding of antibodies against high and low molecular weight cytokeratin proteins in the human placenta with special reference to infarcts, proliferation and differentiation processes.

Recent immunocytochemical studies have shown that placental villous trophoblasts contain the high molecular weight cytokeratin (CK) proteins 5/6 and 17. In the case of CK 17, trophoblastic immunostaining was positive in villi covered by fibrinoid. CKs 5/6 and 17 are expressed by hyperproliferative cells. The aim of this investigation was to examine the location of these CKs in placental infarcts, known to be demarcated by fibrinoid and hyperproliferative trophoblasts. The results were compared with those obtained by immunostaining against Ki-67, tenascin and alpha 1-, alpha 6- and beta 1-integrins, which are involved in cell proliferation, differentiation and regenerative processes. Furthermore, the expression of the single CKs 7, 8, 10, 13, 14, 18 and 19 was investigated by immunocytochemistry and immunoblotting. While low and high molecular weight CKs were present in villous and extravillous trophoblasts, only low molecular weight CKs were detected in vascular and extravascular placental smooth muscle cells. Placental infarcts revealed different immunoreactivities in the infarct margin and centre: high molecular CKs, tenascin, Ki-67 and oncofoetal fibronectin predominated in the infarct margin, low molecular CKs, fibrin and integrins in the centre. The expression of tenascin and a defined change in the expression of CK 17 indicates villous repair and hyperproliferative mechanisms in placental infarcts.

Interleukin-8 in urine: a new diagnostic parameter for intra-amniotic infection after premature rupture of the membranes.

Interleukin-8 (IL-8), a 72 amino acid peptide secreted by cells of the immune system and of the amnion, chorion and decidua, was measured in women in late pregnancy. IL-8 was detected in the urine of 91 of 104 women with premature rupture of the fetal membranes, with values exceeding 1000 ng/L in cases of severe intra-amniotic infection. Women with urinary tract infections were excluded. The routine measurement of IL-8 in urine, together with C-reactive protein in serum, thus provides a low risk and technically simple approach to the assessment of intra-amniotic infection.

Altered urinary interleukin-8/creatinine ratio in peptic ulcer disease: pathological and diagnostic implications.

OBJECTIVES AND METHODS: To assess its clinicopathological and diagnostic significance, interleukin-8 (IL-8) was measured by radioimmunoassay in fasting urine specimens and in gastric mucosal incubates taken from 54 patients with dyspepsia. The presence of Helicobacter pylori, the activity of gastritis, and urine creatinine levels were also assessed. RESULTS: The median urinary IL-8/creatinine ratio was 0.1 x 10(-6) in patients with current peptic ulcers (n = 13) and 0.2 x 10(-6) in patients with a history of ulcers (n = 8), compared with 0.4 x 10(-6) (p < 0.0001) in patients without ulcers who were infected with H. pylori (n = 20) or not infected (n = 13). The activity of gastritis had a positive correlation with gastric IL-8 (r = 0.5870, p < 0.01) and a negative correlation with urinary IL-8/creatinine ratio (r = -0.6447, p < 0.005). The improvement in the activity of gastritis in 20 patients given anti-H. pylori triple therapy was associated with a significant fall in gastric mucosal IL-8 and a rise in urinary IL-8/creatinine ratio. CONCLUSIONS: An inverse relationship seems to exist between urinary IL-8 and the activity of gastritis and gastric IL-8. This may represent another concept in the pathogenesis of peptic ulcers and can assist in the noninvasive diagnosis of peptic ulcer disease.

Histochemical evaluation of placental angiotensinase A in pre-eclampsia: enzyme activity in villous trophoblast indicates an enhanced likelihood of gestational proteinuric hypertension.

The aim of this study was to examine whether differences in placental angiotensinase A (glutamyl aminopeptidase, EC 3.4.11.7) activities occurred in hypertensive complications of pregnancy compared with uncomplicated pregnancies. Biochemical and semiquantitative histochemical methods were used and compared for their applicability. Angiotensinase A activity was detected using L-alpha glutamyl-4-methoxy-2-naphthylamide (alpha-Glu-MNA) as substrate and Fast Blue B salt for simultaneous azo-coupling in cryostat sections of placental tissue samples from 32 patients with pre-eclampsia, 11 patients with pregnancy-induced hypertension and 44 participants with uncomplicated pregnancies. The graduated intensity of reaction product in the villous trophoblast and in fetal blood vessels was evaluated semiquantitatively in a double-blind study by light microscopy (semiquantitative score method). Score levels were related to relative frequencies of hypertensive disorders (proportional odds model) and correlated to the severity of gestational hypertension (Spearman's rank correlation). After detection of enzyme activity, the same tissue samples were homogenized and used for kinetic fluorometric measurements under the same substrate and buffer conditions as in enzyme histochemistry. Enhanced villous trophoblastic angiotensinase A activity was significantly associated with an increased frequency of pre-eclampsia in pregnant women (cumulative odds ratio x 0(1) 6.37; P < 0.001) and showed significant correlations with the severity of gestational hypertensive disorders, represented by systolic (r = 0.31; P < 0.05) and diastolic (r = 0.34; P < 0.05 blood pressure and by concomitant proteinuria (r = 044; P < 0.01). Histochemical evaluation of fetal blood vessels and biochemical measurements revealed no statistically significant results. In conclusion this study demonstrates for the first time that increased villous trophoblastic angiotensinase A activity indicates an increased likelihood of the presence of pre-eclampsia and the severity of hypertensive disorders in pregnancy.

Successful emergency splenectomy during pregnancy in a patient with life-threatening idiopathic thrombocytopenia. Case report.

Thrombocytopenia can be a pathophysiological feature of pregnancy. In the case reported, the thrombocyte count was reduced to 1% of normal (1 x 10(9) thrombocytes/l) at 28 weeks of gestation. In chronological order, the patient showed epistaxis, macrohematuria and gingival, conjunctival, intracerebral and pulmonary bleeding. The latter was life-threatening. An emergency splenectomy was undertaken, without complications. The operation was followed by a massive increase in the thrombocyte count, reaching 200 x 10(9)/l four days later. Unfortunately, a premature rupture of the membranes, with signs of amnion infection, occurred on the seventh day. A Cesarean section was undertaken (30 weeks of gestation), without complications. Both mother and baby are in good health 10 months later. The newborn had a normal thrombocyte count at delivery and thereafter. The life-threatening hemorrhage of the mother, the delivery of an unaffected baby and the relatively quick remission after splenectomy suggest an upregulated destruction of thrombocytes by the maternal spleen. The increased level of Macrophage-Colony Stimulating Factor (M-CSF), a normal feature of pregnancy, has the potential to augment thrombocyte destruction by activating macrophages. The production of anti-thrombocyte antibodies, especially if localized in the spleen, could result in increased thrombocyte sequestration by macrophages with severe effects focused on the mother.

Antibodies to cytokeratins bind to epitopes in human uterine smooth muscle cells in normal and pathological pregnancies.

Cytokeratin antibodies have been widely used for the identification of trophoblast cells in the placental bed, following their invasion from the developing conceptus. Their identification centres upon the expression of cytokeratin in epithelial cells, from which trophoblast cells are derived. Our recent observations indicate that this strict relationship may be more complex than was thought. Cryostat and paraffin sections of human decidua and myometrium, taken from the placental bed and the uterotomy cut, were examined immunocytochemically for cytokeratins using ten antibody clones selected to identify different cytokeratin proteins and antigenic epitopes. Biopsy specimens were obtained from normal and pathological pregnancies (pre-eclampsia, fetal retardation, amnioninfection, hysterorrhexis, placenta praevia) at the time of caesarean section (26-41 weeks of pregnancy). Antibodies against nine clones, CAM 5.2, MNF 116, AE1/AE3, CK5, KS-B17.2, CY-90, M20, E3, and 34 beta E12 identified, as expected, syncytial giant cells and mononuclear trophoblasts within the placental bed and glandular epithelial cells throughout the uterus. In addition, they stained numerous fusiform cells that were classified by established criteria to represent smooth muscle cells, both within blood vessels and myometrium. No staining differences were observed between normal and pathological disorders. These results indicate that cytokeratin antibodies CAM 5.2, MNF 116 and AE1/AE3, and other antibodies targeting proteins 8 and 18, cross-react with epitopes expressed in cells other than giant trophoblastic cells and mononuclear trophoblasts in the uterus and, thus, caution has to be used when such antibodies are used for the diagnostic characterization of tissues related to the placental bed.

[Prenatal diagnosis of fetal polymicrogyria--case report]. / Pränatale Diagnose einer fetalen Polymikrogyrie--Fallbericht.

Migration disorders of foetal neurons are rare conditions which are normally diagnosed after birth and may be followed by severe alterations of neurological development. We describe a case of foetal hemilateral polymicrogyria in combination with hemihypoplasia of the brain, the symptoms of which were diagnosed in the 23rd week of pregnancy, leading to termination of pregnancy. To the best of our knowledge, this is the first case report of antenatal diagnosis of this disorder by vaginal sonography.

[Intrauterine meconium peritonitis. A rare cause of non-immunologic hydrops fetalis]. / Intrauterine Mekoniumperitonitis. Eine seltene Ursache des nichtimmunologischen Hydrops fetalis.

A case of foetal non-immune hydrops fetalis is presented. A second gravida was referred to our department at 31 weeks' gestation after normal pregnancy course. Sonography revealed marked foetal ascites as well as a polyhydramnion. Prenatal examinations did not yield a safe diagnosis. The foetal ascites was punctured via a pigtail-catheter that was left in the foetal abdomen. This procedure was supposed to facilitate the development of foetal lungs. At 33 weeks' gestation, after spontaneous rupture of membranes, a boy was spontaneously delivered. Postpartal radiography pointed to bowel perforation. Surgery on the first day of life showed a severe meconium peritonitis following a perforation of the ileum. 22 months after this operation, the boy is physically and mentally well developed. Until now, there is no sign of mucoviscidosis. We believe that early diagnosis, the possibility of prenatal therapy and the close collaboration of disciplines made the survival of this child possible.

Matrix production of smooth muscle cells from rat aorta in vitro.

Immunofluorescence microscopic methods served to demonstrate the production of the following matrix components in cultures of vascular smooth muscle cells from rat aorta: fibronectin; nidogen; heparan sulphate-proteoglycan (HS-PG); laminin; and collagen types I, III, IV, V, and VI. A time-dependence of synthesis and secretion could be shown for a number of components of the extracellular matrix (ECM), such as laminin. The results revealed the following estimated quantitative differences of the collagen types: type I > type III > types V and VI. A filamentous/fibrillar matrix and also occasionally a typical basal lamina could be demonstrated electron microscopically around the smooth muscle cells.

Electrical and morphological evidence for heterogeneous populations of cultured bovine trabecular meshwork cells.

Although trabecular meshwork cells are presumed to play an important role in determining ocular aqueous outflow resistance, little is known about their membrane transport characteristics. As in vivo access by microelectrodes is difficult, we used cell culture techniques to facilitate membrane voltage recording from cultured bovine trabecular meshwork cells. Phase-contrast microscopy revealed the presence of epithelial-like and spindle-shaped cell types. The mean membrane voltage for epithelial cells was -49.7 +/- 0.8 mV (S.E.M., n = 143) and for spindle cells was -70.9 +/- 1.9 mV (S.E.M., n = 48). These cells possess an electrogenic Na+/K(+)-ATPase and a K+ conductance. K transference numbers (tk) for [K+] from 5 to 80 mM were 0.50 for epithelial cells and 0.71 for spindle cells. The epithelial cells lack the electrogenic Na+/HCO3-symport, thereby enabling their differentiation from corneal endothelial cells and confirming previous reports of differences between these cell types. A proportion of spindle cells demonstrated spontaneous and induced fluctuations of membrane voltage. One millimolar Ba2+ (n = 9) induced an immediate depolarization of membrane voltage, with the onset of 'overshooting' action potentials, which were dependent on extracellular Ca2+ and Na+ but were not blocked by tetrodotoxin, 10(-6) M. Spindle cells showed parallel alignment of intracellular smooth muscle specific alpha-isoactin filaments, whereas epithelial cells showed specks of non-fibrous staining. Electron microscopy revealed that epithelial cells had the characteristics of metabolically active cells, with few intermediate filaments (10-12 nm) and microfilaments (6-7 nm) and short cytoplasmic processes. Spindle cells had long cytoplasmic processes and abundant intermediate- and microfilaments. These data provide further evidence for multiple bovine trabecular cell types. The smooth muscle-like spindle cell may represent the previously proposed contractile element of the angle and its action could conceivably alter ocular outflow resistance.

Membrane voltage recordings in a cell line derived from human ciliary muscle.

A smooth muscle cell line (H7CM) was established from the ciliary muscle of a 1-day-old human infant. The cultured cells had a normal female karyotype (46 XX) and could be maintained in cell culture for at least 11 generations. A common feature of confluent cultures was the presence of abundant bundles of 6-7 nm microfilaments associated with dense bodies. Both the ultrastructural appearance and the presence of smooth muscle-specific alpha-isoactin (also present in the human ciliary muscle in situ) support the smooth muscle origin of the H7CM cell line. Continuous membrane voltage (Vm) recordings were obtained in confluent monolayers of H7CM cells using glass microelectrodes. Resting Vm in 105 impalements averaged -66.2 +/- 0.7 mV (mean +/- standard error of the mean). In this system, rapid membrane transients induced by changing of the superfusing test solutions were detectable. Relative K+ conductance was characterized, and the contribution of electrogenic sodium/potassium adenosine triphosphatase to Vm was investigated. Under control conditions, H7CM cells were electrically quiescent. However, action potentials could be induced by application of 10 mM barium. Barium-induced action potentials were not abolished by removal of extracellular Na+ nor were they inhibited by the presence of tetrodotoxin. However, they were blocked by verapamil, fulfilling criteria believed to be typical for smooth muscle cells. Acetylcholine, carbachol, and to a lesser extent pilocarpine induced a reversible Vm depolarization. The effect of acetylcholine was blocked by atropine, implying muscarinic receptor involvement in the Vm response. Collectively, these findings show the potential usefulness of cultured ciliary muscle cells in understanding further the cellular mechanisms underlying drug-induced contraction of the human ciliary muscle.

Cervical ripening of the rat in dependence on endocrine milieu; effects of antigestagens.

The circumstances which cause the ripening of the cervix during the initial phase of parturition are still unexplored. In addition, complications during labour such as cervical dystocia still necessitate cesarian section. Therefore we examined the biomechanical alterations of the cervix during the last trimester of pregnancy. We developed a special method to measure dilatation and resistance of the rat cervix. After testing several substances, the antigestagen Mifepristone turned out to be most effective in causing cervical softening. Further investigations with light and scanning electron microscopy showed an increase of collagen and a possible loosening of collagen fibers after antigestagen treatment. Therefore it might be possible to use Mifepristone, perhaps in combination with a prostaglandin to promote cervical ripening when certain obstetric problems arise.

Morphological evaluation of steatosis in monolayer cultures (MDCK cells) after treatment with gentamicin and valproic acid.

An attempt was made to develop a method for the morphological and morphometric evaluation of fat inclusions in monolayer cell cultures. For this purpose we used electron microscopy (SEM and TEM) and Sudan black staining methods followed by morphometric evaluation techniques. MDCK (Madin-Darby-canine-kidney) cells were treated with valproic acid (VPA; 10, 100, 1000 micrograms/ml medium for 22 hrs) and gentamicin (1, 10, 50 micrograms/ml for 60 min). Sudan black staining revealed a dose-dependent increase in fat deposits after application of the two substances. SEM inspection showed differences in the size and distribution of the fat inclusions. TEM findings only partly allowed us to differentiate clearly between VPA- and gentamicin-induced fat inclusions.