bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of Electrical Conductivity of Amniotic Fluid in the Evaluation of Fetal Lung Maturation
Authors: Pachì A. , De Luca F. , Cametti C. , Barresi S. , Berta S.
Published: April 2003

The aim of this work is to evaluate, by means of an alternative method, the fetal lung maturation by means of measurement of the electrical conductivity of the amniotic fluid correlated with the phospholipid content. In vitro measurements were made in some 100 amniotic fluid samples at different gestational stages. A considerable change was observed in the electrical conductivity of the amniotic fluid in the last period of pregnancy, which reflects the increase in phospholipid concentration, when lung maturation is reached. We hope that further investigations, on wider case histories, will enable the application in vivo of the above-mentioned methodology.
Chromosomal Mosaicism for Isochromosome 11q Confined to CVS Direct Preparations
Authors: Sanz R. , Lorda-Sanchez I. , Fernández-Moya J.M. , Cifuentes-Sulzberger S. , Rodríguez de Alba M. , Gonzalez-Gonzalez M.C. , Ibañez M.A. , Robledo M. , Ayuso C. , Ramos C.
Published: April 2003

Objective: To analyse the discrepancy between the karyotype in direct preparations of chorionic villus sampling (CVS) and the fetal karyotype and its possible fetal phenotypic repercussion. Methods: The karyotype was obtained from direct and cultured preparations of CVS. FISH was performed in direct CVS preparations and in four different areas of term placenta. Results: Karyotype and FISH analysis in CVS revealed a 46,XX/47,XX,+i(11q) cell line. Cultured CVS preparations showed a 46,XX karyotype. Cytogenetic studies in term placenta did not reveal the abnormal cell line. Molecular studies did not detect uniparental disomy for chromosome 11 in the fetus. Conclusion: The fetus, at birth, had no phenotypic abnormalities. IUGR was not present during gestation, in accordance with the low proportion of aneuploid cells in term placenta, and UPD for chromosome 11 was not observed.
Short Rib-Polydactyly SyndromeType III: Comparison of Ultrasound, Radiology, and Pathology Findings
Authors: Golombeck Kirstin , Jacobs Volker R. , von Kaisenberg Constantin , Oppermann Hans-Conrad , Reinecke-Lüthge Axel , Weisner Dietrich , Jonat Walter
Published: April 2003

Short rib-polydactyly syndrome (SRPS; types I–IV) is an autosomal recessive, lethal skeletal dysplasia characterized by short-limb dysplasia, narrow thorax, and polydactyly. This syndrome is invariable and can be detected by 2-trimester ultrasound. The underlying gene has not been discovered yet. We report a case of SRPS subtype III Verma-Naumoff-Le Marec that was sonographically detected at 20 weeks’ gestation and compare prenatal ultrasound with postmortem findings from pathology and radiology. Since the risk of recurrence is 25%, early ultrasound for consecutive pregnancies was advised and performed at 11+6 weeks’ gestation in the following pregnancy without any findings. Ultrasound diagnosis in this rare case of SRPS is a valuable tool for identification and early management, since there are no specific biochemical or histopathological markers for this syndrome. Radiological and pathological findings confirmed SRPS type III and assisted in the differential diagnosis of the subtype.
First Trimester Diagnosis of Split Hand/Foot by Transvaginal Ultrasound
Authors: Haak M.C. , Cobben J.M. , van Vugt J.M.G.
Published: April 2003

Objective/Method: First trimester high-frequency transvaginal ultrasonographic examination was performed in a 26-year-old gravida 2, para 1, diagnosed with tetramelic split hands and feet. The couple was informed about the probable 50% recurrence risk of this anomaly. Results: At 12 weeks’ gestation, high-frequency transvaginal ultrasound examination revealed a fetus with biometry consistent with gestational age. Both hands appeared to have a single digit at the ulnar side. On the site of the metacarpal bones, only three bones could be visualized. Both feet showed two digits, with a typical ‘split-foot’ appearance. Nuchal translucency thickness measured 1.0 mm. No additional abnormalities could be found. The child was born in good condition with the described split hand/foot malformation (SHFM). Conclusion: This case report demonstrates prenatal sonographic diagnosis of SHFM in a first trimester fetus. In first trimester there can be several technical advantages and it shortens the period of uncertainty of the parents. Secondly it provides the option of early surgical termination of the pregnancy instead of induction of labour.
Different Policies on Prenatal Ultrasound Screening Programmes and Induced Abortions Explain Regional Variations in Infant Mortality with Congenital Malformations
Authors: Garne Ester , Berghold Andrea , Johnson Zachary , Stoll Claude
Published: April 2003

Objective: To compare the impact of induced abortions (IA) on the mortality of infants with congenital malformations in four European regions with different policies on IA and prenatal ultrasound screening for congenital malformations. Methods: A registry-based collection of data on congenital malformations in four different countries: Ireland (Dublin), Denmark (Funen County), Austria (Styria), and France (Strasbourg). Results: The proportion of infant deaths with malformations ranged from 23 to 44% of all infant deaths with the highest proportion in Dublin, where IA is not allowed and prenatal ultrasound screening not performed. There were highly significant differences in the prevalences of IA (p < 0.001), fetal deaths (p < 0.01), and deaths in infants with congenital malformations (p < 0.001) between the four regions. The differences in total mortality with congenital malformations (IA + fetal deaths + infant deaths) between regions decreased, and only Strasbourg differs significantly from the other three regions. Conclusion: Prenatal ultrasound screening programmes have only a minor impact on total mortality with congenital malformations from 2nd trimester of pregnancy to 1 year of age, but seem to change the time of death which may be important for both the parents and the community.
Maternal Serum CA125 and CA15-3 Antigen Levels in Normal and Pathological Pregnancy
Authors: Bon G.G. , Kenemans P. , Verstraeten A.A. , Go S. , Philipi P.A. , van Kamp G.J. , van Geijn H.P. , van Vugt J.M.G.
Published: April 2003

Objective: To evaluate the value of maternal serum CA125 and CA15-3 concentrations for discriminating pathological from normal pregnancies. Methods: Serum samples from 120 women, in whom pregnancy outcome was pathological, i.e. spontaneous abortion, fetal death, intrauterine growth retardation, chromosomal and structural abnormalities, and (pre)eclampsia, were assessed for CA125 and CA15-3 and compared with levels found in 350 women with a normal pregnancy outcome matched for age and duration of pregnancy. Results: Maternal CA125 serum values were significantly higher in the first and the third trimester of pregnancy (median 23.0 and 21.0 U/ml; p < 0.00001 and p < 0.001, respectively), compared to those in the second trimester (median 14.0 U/ml), but not significantly different from those obtained in pathological pregnancies. Maternal serum CA15-3 values were significantly higher during the third trimester (median 26.0 U/ml) compared to the first and second trimester of pregnancy (median 14.0 and 15.0 U/ml; p < 0.0001); CA15-3 serum levels in normal and pathological pregnancies showed no significant difference. Conclusion: Maternal serum levels of CA125 are higher during the first and third trimester of pregnancy. CA15-3 maternal serum levels are higher during the third trimester compared to the first and second trimester. Maternal CA125 and CA15-3 serum levels showed no relation with a pathological outcome of pregnancy.
Prenatal Detection of Facial Clefts
Authors: Sohan Karen , Freer Mandy , Mercer Nigel , Soothill Peter , Kyle Phillipa
Published: April 2003

Objectives: To determine (1) the antenatal detection rate for isolated cleft lip and/or cleft palate during the routine anomaly scan; (2) the correlation between prenatal diagnosis and postnatal findings, and (3) the association of apparently isolated cleft lip and/or cleft palate with other anomalies, in particular chromosomal abnormalities. Method: A population-based retrospective analysis of all cases of isolated cleft lip and/or cleft during an 8-year period in an academic teaching hospital in the UK. Results: Thirty-nine cases of isolated cleft lip and/or cleft palate were identified among deliveries at the hospital. Twenty-eight cases had a routine anomaly scan. Fourteen cases were detected prenatally (sensitivity 50%). None of the isolated cleft palates was detected, while 14 of 20 cases of cleft lip (70%) were detected. One of the isolated cases of cleft lip was associated with trisomy 21, while 3 of the isolated cleft palate cases were associated with the Pierre Robin syndrome. In all cases, an antenatal diagnosis of cleft was confirmed following delivery or post-mortem examination (specificity 100%). Conclusions: Ultrasound is a useful tool in screening for cleft lip with or without cleft palate, but not for cleft palate alone. Even with an isolated cleft lip, there is an increased risk of chromosomal abnormality. The role of prenatal education and support is extremely important in the preparation of prospective parents and can help alleviate the shock which occurs when there is an unexpected cleft at birth.
Persistent Detection of Alfa-Fetoprotein in the Vagina without Overt Preterm Premature Rupture of the Membranes
Authors: Kishida Tatsuro , Yamada Hideto , Furuta Itsuko , Kobayashi Noriko , Hirayama Emi K. , Ebina Yasuhiko , Cho Kazutoshi , Okuyama Kazuhiko , Fujimoto Seiichiro
Published: April 2003

Objectives: The objectives of this study were to clinically characterize and seek risk factors for the patients with persistently detected alfa-fetoprotein (AFP) in the vagina without showing overt preterm premature rupture of the membranes (PROM). Study Design: The cohort study was undertaken using vaginal-cervical specimens collected consecutively between 21 and 40 weeks of gestation from 25 consenting women. Demographic, obstetric, neonatal, and laboratory data from patients with persistent positive results of AFP kit tests without showing overt leakage of amniotic fluid were compared to those from controls. Cervical levels of interleukin-1α, interleukin-1β, interleukin-6, interleukin-8, and granulocyte elastase were measured by immunoassay, and the uterine cervix was assessed by transvaginal ultrasonography. Statistical analysis involved Fisher’s exact test and Mann-Whitney U test. Results: The incidence of overt preterm PROM was significantly higher in the patients with persistently detected AFP in preterm (3 in 4 cases) than in controls (3 in 21 controls). At sampling before determination of persistently detected AFP, patients with persistently detected AFP had significantly higher cervical levels of interleukin-6 and significantly shorter cervical length than controls. Conclusions: Patients with persistently detected AFP in the vagina without showing overt preterm PROM have the risk of overt preterm PROM. Increased levels of interleukin-6 in cervical specimens and short cervical length may be risk factors for the occurrence of persistent detection of AFP in the preterm.
Mild Ventriculomegaly, Mild Cerebellar Hypoplasia and Dysplastic Choroid Plexus as Early Prenatal Signs of CHARGE Association
Authors: Becker Rolf , Stiemer Beate , Neumann Luitgard , Entezami Michael
Published: April 2003

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.
Biochemical Differentiation of Gestational Compartments in the Midgestational Fetal Rabbit
Authors: Devlieger R. , Gratacós E. , Wu J. , Ardon H. , Vereecken A. , Deprest J.
Published: April 2003

Objectives: The fetal rabbit at midgestation is increasingly being used as a model in fetal diagnosis and therapy. In this study, we aimed to establish a reliable method for identification of the origin of sampled extra-embryonic fluids based on selected biochemical components. Methods: In 6 pregnant does at 22 days of gestation, 18 gestational sacs were sampled for amniotic, allantoic and exocoelomic fluid. These fluids, as well as matching maternal and fetal blood samples, were assayed for levels of sodium, potassium, chloride, bicarbonate, total protein, alkaline phosphatase, γ-glutamyl transferase and progesterone. Results: Levels of sodium and potassium were, respectively, lower and higher in the allantoic fluid when compared to other extra-embryonic spaces. Amniotic fluid had a significantly lower total protein content and higher level of alkaline phosphatase when compared to the exocoelomic fluid. Significant levels of progesterone could only be detected in maternal blood. Conclusions: In the midgestational rabbit, a combined assay of potassium, alkaline phosphatase and progesterone can determine the gestational cavity of origin of the sampled fluid. The obtained gradients for these markers suggest compartment-specific production and/or inter-cavity transfer mechanisms.
Management of Anti-Rhesus-D Antibodies in Pregnancy: A Review from 1994 to 1998
Authors: Cheong Ying C. , Goodrick Jean , Kyle Phillippa M. , Soothill Peter
Published: April 2003

Objective: To review our management of anti-Rhesus-D antibodies in pregnancy over a 5-year period in order to assess possible changes in the management or prognosis which may have developed with time. Method: Retrospective analysis of prospectively collected data from 31 pregnancies with maternal anti-D levels >4 IU/ml and in which the fetus was Rhesus positive. Results: There were a total of 30 amniocenteses, 8 cordocenteses, and 54 fetal blood transfusions performed. When undertaken as the first procedure, the mean gestational age at amniocentesis was 30 weeks as compared with 25 weeks for fetal blood sampling/transfusion (p < 0.05). The median anti-D level at the first procedure was 24 IU/ml for amniocentesis and 64 IU/ml for fetal blood sampling. Of the 54 blood transfusions, 43 were intravascular, 4 were intraperitoneal, and 7 transfusions were both intravascular and intraperitoneal. Conclusions: Intravascular as opposed to intraperitoneal transfusions were found to be the main method of transfusion in the later years in this study, a finding which was expected with improved sonographic equipment. Apart from this, management and prognosis of anti-D red cell isoimmunisation in pregnancy have remained relatively stable since the 1980s. Amniocentesis was useful in the management of such pregnancies, especially as an initial procedure in the cases with a lower initial anti-D level. In this series 90% of the fetuses requiring blood transfusion, but were without hydrops, survived, whereas this was about 70%, if they had become hydropic (this latter figure was reduced by 2 hydropic deaths before 20 weeks’ gestation in the same very severely affected woman).
High-Efficiency Retroviral Transduction of Fetal Liver CD38–CD34++ Cells: Implications for in utero and ex utero Gene Therapy
Authors: Ohkubo Tatsuo , Bárcena Alicia , Smith Clayton A. , Harrison Michael R. , Muench Marcus O.
Published: April 2003

Objectives: Defining methods for the efficient transduction of fetal stem cells could lead to novel fetal therapies for blood cell disorders and other birth defects. In this study, we analyzed the effects of various parameters on the retroviral transduction of primitive hematopoietic progenitors/stem cells isolated from fetal liver. Methods: Candidate stem cells were isolated by fluorescence-activated cell sorting from midtrimester human livers based on the phenotype CD38–CD34++lineage– (lineage = glycophorin A, CD3, CD14, CD19, CD20 and CD56). A murine retroviral vector with a truncated human low-affinity nerve growth factor receptor (ΔNGFR) gene was used to transduce the candidate stem cells. Marker gene expression was monitored by flow cytometry using an anti-NGFR mAb. Candidate stem cells were transduced immediately after isolation or after up to 4 days of culture in serum-deprived medium containing the growth factors kit ligand and granulocyte-macrophage colony-stimulating factor. The effects on transduction efficiency of the addition of 4 µg/ml protamine sulfate and/or centrifugation to concentrate the candidate stem cells and virus were tested. After transduction, the cells were expanded for 10–21 days before determining the frequency of NGFR+ cells among the different hematopoietic progeny. Results: Efficient transduction of candidate stem cells, at an average rate of 46%, was achieved after 3 days of culture with a single exposure to virus. Longer than 3 days of culture or repeated exposure to viral supernatant did not significantly improve the rate of transduction. The use of centrifugation at 1,200 g for 1 h and the addition of protamine sulfate during the transduction procedure were critical to achieving a high rate of transduction. Marker gene expression was observed on the progeny of the transduced cells in conjunction with CD34 (progenitors), glycophorin A (erythrocytes), CD14 (monocytes), CD15 (granulocytes) and CD41 (megakaryocytes). Conclusions: This study demonstrates that the efficient transduction of fetal candidate stem cells can be achieved under defined culture conditions using a retroviral vector. These results encourage further examination of in utero and ex utero gene therapy as a means of treating birth defects.
The Rationale for in utero Repair of Myelomeningocele
Authors: Walsh Danielle S. , Adzick N. Scott , Sutton Leslie N. , Johnson Mark P.
Published: April 2003

Objectives: Despite advances in prenatal diagnosis, management of fetal myelomeningocele has been limited to abortion or supportive postnatal care. The rationale for fetal repair of myelomeningocele and initial clinical outcomes are discussed. Methods: A complete review of the literature concerning fetal myelomeningocele and repair was performed. Results: While myelomeningocele is a primary embryologic disorder, neurologic damage is also secondary to progressive in utero damage to the exposed spinal cord. Animal models with midgestational coverage of the spinal defect demonstrate near normal neurologic function at term. Early clinical results suggest that fetal closure can salvage neurologic function, reverse hindbrain herniation, and diminish the need for ventriculoperitoneal shunting. Conclusions: In utero repair of myelomeningocele may improve neurologic outcomes and reduce hindbrain herniation in selected patients.
Prenatally Diagnosed Microcephaly: A Review of Etiologies
Authors: Dahlgren Leanne , Wilson R. Douglas
Published: April 2003

Objective: To determine the incidence and etiology of prenatally diagnosed microcephaly. Methods: Retrospective review of 10 years at a tertiary obstetrical hospital. The study population consisted of 21 infants with confirmed prenatal and postnatal microcephaly. Results: There were 8 different definite/probable etiologies identified (viral, monochorionic twinning, genetic syndrome, neural tube defect, abnormal karyotype, hypoxic insult, constitutional). Conclusions: Retrospective study indicates the commonest etiologies for prenatal microcephaly are in utero infection, monochorionic twin insult, rare genetic syndrome and chromosomal anomalies.
Doppler Ultrasonography and Implantation: A Critical Review
Authors: Carbillon L. , Perrot N. , Uzan M. , Uzan S.
Published: April 2003

Active perifollicular angiogenesis in ovaries and endometrium is linked with the quality of oocytes and endometrial receptivity, and consequently with the outcome of implantation. Studies evaluating ovarian velocimetry in relation to implantation and pregnancy rates bring essentially into play velocity values or non-quantitative parameters, whose reproducibility is low and which are hardly usable in daily practice. Conversely, the assessment of uterine artery flow by Doppler examination is easy and reproducible, and high impedance at the end of the follicular phase is a good predictive indicator of poor endometrial receptivity. In early pregnancy, Doppler investigation must not be used routinely because its safety is still under discussion; however, the available data show that impedance in the uterine and spiral arteries does not show any significant difference between normal and abnormal pregnancies (missed abortions or anembryonic pregnancies), and the significance of easily detectable intervillous flow is hotly debated, raising fundamental points.
Fetal Autopsy: The Most Important Contribution of Pathology in a Center for Perinatal Medicine
Authors: Tennstedt Cornelia , Hufnagl P. , Körner H. , Vogel M. , Dietel M. , Chaoui R.
Published: April 2003

Objective: The aim of the current paper is to analyze the significant changes in fetal autopsy made by pathologists in a center for perinatal medicine in response to the recent rapid developments in prenatal medicine. Methods: We use our technical equipment (interdisciplinary databank, Intranet, fine instruments, imaging techniques) for prenatal diagnostics and fetal pathology in the Center for Perinatal Medicine at the Charité University Medical Center in Berlin, Germany. Results: Our technical setup has made it possible for us to plan autopsies in such a way that rare malformations in special cases can be documented in sectional planes corresponding to the prenatal sonographic images. Before the postmortem examination the pathologist receives all the prenatal findings obtained for the fetus and is able to examine the prenatal images on a monitor. The final diagnosis in the postmortem report includes the results of modern molecular pathology, clinical-genetic examination of the fetus, cytogenetic and molecular-cytogenetic findings and postmortem x-rays. Conclusions: To be able to incorporate these various results into a final report in routine work the use of an interdisciplinary database is essential. Such a database also ensures faster and more productive interdisciplinary communication in a center of perinatal medicine.
Large Placental Chorioangioma as a Cause of Sudden Intrauterine Fetal Death
Authors: Batukan Cem , Holzgreve Wolfgang , Danzer Enrico , Bruder Elisabeth , Hösli Irene , Tercanli Sevgi
Published: April 2003

A giant placental chorioangioma was diagnosed in a fetus at 22 weeks of gestation by prenatal ultrasound screening and color Doppler imaging. Although no signs of fetal hydrops and cardiac decompensation were observed, the situation of the fetus deteriorated rapidly and in utero fetal death occurred at 26 weeks, attributed to bleeding from ruptured sinusoids within the tumor. Prenatal diagnosis of chorioangioma and treatment modalities are discussed.
Accuracy of Prenatal Diagnosis of Congenital Heart Defects
Authors: Berghella Vincenzo , Pagotto Luciana , Kaufman Marion , Huhta James C. , Wapner Ronald J.
Published: April 2003

Objectives: (1) To evaluate the detection and accuracy of fetal echocardiography for congenital heart defects (CHD). (2) To compare the detection and accuracy of a team of maternal fetal medicine specialists and radiologists (MFM/R) with those of perinatal cardiologists (PC). Methods: All fetal echocardiograms performed from 1/91 to 6/95 were reviewed retrospectively. CHD diagnoses made prenatally were compared with postnatally-confirmed diagnoses. Results: 106 heat defects were correctly detected prenatally. There were 10 incorrect diagnoses, 6 false-negatives and 4 false-positives. Overall detection rate (sensitivity) was 95%, while overall accuracy was 87%. Detection rate for MFM/R and PC were 91 and 97%, respectively (p = 0.4). Accuracy was 74% for MFM/R and 92% for PC (p = 0.02). Conclusions: CHD can be identified reliably by prenatal echocardiography. The involvement of a PC in the prenatal diagnosis of these defects may improve diagnostic accuracy.
Prenatal Echocardiographic Appearance of Arrhythmogenic Right Ventricle Dysplasia: A Case Report
Authors: Rustico M.A. , Benettoni A. , Fontaliran F. , Fontaine G.
Published: April 2003

We report a case of arrhythmogenic right ventricular dysplasia (ARVD) diagnosed prenatally by echocardiography at 24 weeks gestation. The 4-chamber view showed a large outpouched area extending from below the tricuspid valve to the insertion of the moderator band; the affected wall appeared thin and akinetic, with absence of flow at color Doppler investigation and no evidence of cardiovascular failure. The size of the outpouched area was unchanged at subsequent controls (25 and 26 weeks gestation) when frequent extrasystoles occurred, probably of a ventricular origin. The pregnancy was terminated at 27 weeks. The histopathologic examination of the fetal heart showed the presence of clusters of adipocytes interspersed with myocardial fibers, consistent with the diagnosis of ARVD.
DNA-Encoding Enzymatically Active HIV-1 Reverse Transcriptase, but Not the Inactive Mutant, Confers Resistance to Experimental HIV-1 Challenge
Authors: Isaguliants Maria G. , Petrakova Natalia N. , Zuber Bartek , Pokrovskaya Katja , Gizatullin Rinat , Kostyuk Dmitrii A. , Kjerrström Anne , Winberg Gösta , Kochetkov Sergey N. , Hinkula Jorma , Wahren Britta
Published: April 2003

The present study was undertaken to examine the immunogenicity of a single plasmid DNA representing the reverse transcriptase (RT) of HIV-1. Plasmids containing the enzymatically active RT as well as a mutated nonenzymatically active RT with nucleotide (nt)-binding motifs of YMDD and YMLL, respectively, were used to immunize mice. Both constructs induced similar good antibody and T cell responses, with a tendency towards antibody directed to peptides representing the active and mutated sites. Immunized mice were challenged with a murine pseudotype HIV-1/MuLV infected spleen cells. Seven out of 10 mice immunized with RT had no recoverable HIV-1, while 10 individuals immunized with the RT mutant and all the 18 controls had high levels of recoverable HIV-1. This indicates that mutation of RT reduces the desired immunogenicity.
Modern Approach to Diagnosis in Patients with Suspected Deep Vein Thrombosis
Authors: Wells Philip S. , Anderson David R.
Published: April 2003

Deep vein thrombosis is a relatively common disease, amenable to therapy but with a potentially fatal outcome if untreated. The diagnosis can be made in most patients with the non-invasive imaging procedure ultrasonography, but limitations exist. As with all tests, there is a potential for false-positive and false-negative results. The latter are especially an issue for calf vein thrombi, and this in part has led to the concept of serial testing of the proximal venous system rather than imaging the calf. The premise of the repeat (serial) test is that only thrombi that extend to the proximal system are clinically relevant and such thrombi will be detected on the repeat test. However, despite the safety of the serial-testing concept, it is inconvenient and expensive. In the last few years, the diagnostic process has been improved by the validation of a clinical model that accurately categorizes patients as having low, moderate or high probability of deep vein thrombosis. Among the improvements this provides is the elimination of serial testing if the ultrasound is normal and the clinical probability low. The fibrin degradation product D-dimer has been demonstrated to have a high negative predictive value and has also proven useful in diagnostic algorithms. The combination of the D-dimer with clinical model assessment will enable diagnostic-testing strategies that are more safe, effective and convenient for patients.
Use of Glycoprotein IIb/IIIa Inhibitors in Patients with Acute Coronary Syndromes Undergoing Coronary Revascularization
Authors: Popma Jeffrey J.
Published: April 2003

This paper briefly reviews the results from three recent large-scale clinical trials evaluating the benefit of glycoprotein IIb/IIIa inhibitors in patients with acute coronary syndromes (ACS). The available data suggest that these are promising agents for the management of ACS, particularly in improving the immediate outcome in the acute setting.
Morphological Differences between GPIb Antibody-Induced and Shear Stress-Induced Platelet Aggregates
Authors: Nomura Shosaku , Tandon Narendra N. , Nakamura Takashi , Kambayashi Junichi
Published: April 2003

We determined the morphological differences between NNKY5-5-induced and shear stress-induced platelet aggregates using confocal laser scanning microscopy, and investigated the effects of cytokines on these aggregates. As shear stress was increased from low to high, many aggregates were generated. Some platelets and aggregates already exhibited PAC-1 binding at low shear stress. And finally, aggregates were clearly stained by PAC-1 at high shear stress. Low shear stress-induced aggregates included fibrinogen, but not von Willebrand factor (vWF). In contrast, high shear stress-induced aggregates included both fibrinogen and vWF. NNKY5-5-induced platelet aggregates exhibited both PAC-1 binding and fibrinogen, but vWF was not found in them. The aggregate formation in NNKY5-5-treated platelet-rich plasma (PRP) at low shear stress was clearly more pronounced than that in untreated PRP. In addition, aggregates in NNKY5-5-treated PRP formed after 6 min under low shear stress were clearly stained by PAC-1 and included fibrinogen, but vWF was not found in them. In order to investigate the effects of cytokines on platelet activation under NNKY5-5 stimulation, changes in light transmission and light scatter were assessed with an AG-10. G-CSF, IL-6 and thrombopoietin (TPO) each enhanced light scatter compared to control PRP, although IL-3, GM-CSF, erythropoietin and stem cell factor did not. In particular, TPO significantly enhanced the ‘%-T’ and ‘S-Max’ of NNKY5-5-treated PRP. TPO clearly enhanced the aggregate formation with high shear stress or NNKY5-5 stimulation. These results suggest that the glycoprotein Ib (GPIb)-unmediated aggregates can be formed with only fibrinogen, but the GPIb-mediated aggregates by vWF and fibrinogen synergistically. In particular, the latter is formed more firmly, and both aggregates are enhanced by TPO.
Antithrombotic Activity of Orally Administered Low Molecular Weight Heparin (Logiparin) in a Rat Model
Authors: Hiebert Linda M. , Ping Tilly , Wice Sandra M.
Published: April 2003

Previous studies in rats demonstrated that orally administered, unfractionated bovine lung heparin is absorbed and has a dose-dependent antithrombotic effect. The objective of this study was to determine if an oral low molecular weight heparin had a similar antithrombotic effect in the same model. Thrombosis was induced in rats by application of 10% formalin in 65% methanol to the exposed jugular vein. Immediately following, saline, unfractionated heparin (3.3–60 mg/kg) or the low molecular weight heparin, Logiparin (0.025–15 mg/kg; 20–30 rats per group) was placed in the stomach and 4 h later the jugular vein was inspected for a thrombus. Compared to saline, oral Logiparin reduced the incidence of thrombosis at all doses with a dose-dependent effect suggested. A significant increase was observed in the activated partial thromboplastin time and in plasma heparin concentrations, determined by AccuclotTM Heptest® and anti-factor Xa chromogenic assay for rats given oral Logiparin versus saline. A dose-dependent increase in plasma heparin concentration was observed when estimated by the anti-Xa chromogenic assay. Heparin was recovered in 9% of aortic endothelial samples when ≧0.8 mg/kg Logiparin was administered. A 50% reduction in thrombosis was observed at 0.1 mg/kg for oral Logiparin versus 7.5 mg/kg for unfractionated bovine lung heparin indicating that oral Logiparin is an effective antithrombotic agent at doses lower than unfractionated heparin. Orally administered low molecular weight heparin may be useful for the prevention and treatment of thrombosis.
Formation, Inhibition and Clearance of Plasmin in vivo
Authors: Chandler W.L. , Alessi M.C. , Aillaud M.F. , Vague P. , Juhan-Vague I.
Published: April 2003

A 5 µg/kg bolus of tissue plasminogen activator (t-PA) was infused into 11 healthy subjects followed by measurement of t-PA activity and antigen, PAI-1 activity and antigen, t-PA/PAI-1 complex, plasmin/antiplasmin (PAP) complex and D-dimer over 4 h. Infusion of t-PA resulted in a rise in PAP levels in all subjects from a baseline of 2.4 ± 1.1 nmol/l to a peak of 5.1 ± 2.3 nmol/l, but had no effect on plasminogen, antiplasmin or D-dimer levels. Using a kinetic model of plasminogen activation in vivo, the second-order rate constant for t-PA binding to plasminogen was estimated to be 3,100 ± 1,300 mol–1l·s–1, 200–500 times slower than t-PA accelerated by fibrin. The half-life of PAP was 4.5 ± 1.6 h. In healthy subjects, the PAP level in plasma represents the average rate of plasminogen activation over the last 2–8 h.
Why Is the Standardization of Prothrombin Time a Problem?
Authors: Talstad Ingebrigt
Published: April 2003

The standardization of prothrombin time (PT) has been a problem for more than 50 years; by applying the methods to the same clinical material the analysis may explain why. The initial standardization by 2–3 times normal clotting time by Quick’s PT assay did not standardize presently used PT assays. The ratio method accepted by the WHO in 1977 was not satisfactory because the intercept was neglected. The revised ratio method (WHO 1983) recommended calibration between log clotting times of normal and abnormal plasmas and a simplified conversion into ratios. However, by including a high proportion of normal plasmas (up to 1/3 of abnormal) an erroneous calibration material was introduced; the conversion from clotting times into ratios was also more complex than predicted. The international sensitivity index, which is based on the simplified method, produced international normalized ratios (INR) that were not more standardized than not corrected ratios. Not corrected coagulation activities showed poor standardization due to the different sensitivity of assays for the protein induced by vitamin K absence (Pivka) inhibitor. The corresponding therapeutic ranges of INR and coagulation activities covered 70 and 10–20% of their scales, respectively. Standardization by Pivka-corrected coagulation activities against an international reference assay may be the preferable method.
Determination of Phylloquinone and Menaquinones in Food
Authors: Schurgers Leon J. , Vermeer Cees
Published: April 2003

Fluctuations in international normalized ratio values are often ascribed to dietary changes in vitamin K intake. Here we present a database with vitamin K1 and K2 contents of a wide variety of food items. K1 was mainly present in green vegetables and plant margarins, K2 in meat, liver, butter, egg yolk, natto, cheese and curd cheese. To investigate the effect of the food matrix on vitamin K bioavailability, 6 healthy male volunteers consumed either a detergent-solubilized K1 (3.5 µmol) or a meal consisting 400 g of spinach (3.5 µmol K1) and 200 g of natto (3.1 µmol K2). The absorption of pure K1 was faster than that of food-bound K vitamins (serum peak values at 4 h vs. 6 h after ingestion). Moreover, circulating K2 concentrations after the consumption of natto were about 10 times higher than those of K1 after eating spinach. It is concluded that the contribution of K2 vitamins (menaquinones) to the human vitamin K status is presently underestimated, and that their potential interference with oral anticoagulant treatment needs to be investigated.
Conjunctive Effects of the 5HT2 Receptor Antagonist, Sarpogrelate, on Thrombolysis with Modified Tissue Plasminogen Activator in Different Laser-Induced Thrombosis Models
Authors: Yamashita T. , Kitamori K. , Hashimoto M. , Watanabe S. , Giddings J.C. , Yamamoto J.
Published: April 2003

The effect of the serotonin (5HT2) receptor antagonist, sarpogrelate, was compared with that of the selective thrombin inhibitor, argatroban, in modified tissue plasminogen activator (mt-PA)-induced thrombolysis using two laser-induced thrombosis models reflecting different levels of vascular endothelial cell damage. Bolus intravenous infusions of mt-PA (0.1, 0.2, 0.4 mg/kg) induced thrombolysis in a dose-dependent manner. Sarpogrelate (4.7 mg/kg b.i. + 1.0 mg/kg/h i.v.) given together with mt-PA (0.2 mg/kg b.i.) optimally enhanced thrombolysis (p < 0.05) in a helium-neon laser-induced model where endothelial damage was minimal but not in an argon laser model where desquamation of endothelial cells was recognized. In contrast, argatroban (0.5 mg/kg b.i. + 0.1 mg/kg/h i.v.) given with mt-PA (0.2 mg/kg b.i.) significantly enhanced thrombolysis in both laser models. The findings indicate that the effectiveness of sarpogrelate in thrombolytic therapy might depend on the extent of vascular damage.
Economics of Hemophilia Care
Authors: Aledort Louis M.
Published: April 2003

//8R Hemophilia care is delivered differently in many parts of the world. Some areas of the world cannot diagnose hemophilia, other recognize the disease but do not have treatment, while others provide comprehensive care and have a plethora of services, as well as unlimited replacement therapy. The common denominator in determining the extent of care is the availability of financial support for it. Today, even wealthy nations which have provided unlimited support for care are seriously questioning or limiting expenditures on high-cost diseases. Hemophilia, a life-long chronic disease, is one which is very costly, and which serves both as a model of chronic disease care, as well as a target for financial scrutiny. Health care providers, in concert with patients and their families, must be able to provide data to ensure continued financial underwriting.
Antiphospholipid Syndrome: State of the Art with Emphasis on Laboratory Evaluation
Authors: Greaves Michael
Published: April 2003

Antiphospholipid antibodies (aPLs) are associated with arterial and venous thrombosis, recurrent pregnancy loss and thrombocytopenia. Although aPLs have not yet been conclusively shown to be causal in thrombosis and miscarriage, they are useful laboratory markers for the antiphospholipid syndrome (APS). The syndrome can complicate another autoimmune disease, most commonly systemic lupus erythematosus, but more often occurs alone – primary APS. Identification of the syndrome is clinically important because of the risk of recurrent thrombosis and the need for antithrombotic therapy in many cases. Diagnosis and treatment of APS represent significant challenges, however, owing to the protean clinical manifestations and associations, limitations of currently available laboratory tests for aPLs, and the lack of clear evidence-based guidance on optimal management.
Physiological Function of Tissue Factor Pathway Inhibitor and Interaction with Heparins
Authors: Sandset Per Morten , Bendz Bjørn , Hansen John-Bjarne
Published: April 2003

Tissue factor pathway inhibitor (TFPI) is now recognized as a major physiological anticoagulant. Its main role is to modulate factor VIIa/tissue factor catalytic activity. Another important role is to potentiate the effect of heparins. TFPI is released from the vascular endothelium after injection of either unfractionated heparin (UFH) or low-molecular-weight heparins (LMWHs), which may then provide high concentrations of TFPI at sites of tissue damage and ongoing thrombosis. In dilute prothrombin-time-based assays, released TFPI contributes approximately one-third to the anticoagulant effect of heparin, the remaining being accounted for by antithrombin. Released TFPI, but not plasma TFPI, contains the basic carboxy-terminal tail which is important for the anticoagulant effect. UFH and LMWH exert differential effects on intravascular TFPI. UFH, but not LMWH, given in therapeutic doses, is associated with a progressive depletion of TFPI, which is associated with a strong rebound activation of coagulation after cessation of treatment. Such depletion may explain the apparent superior efficacy of LMWH observed in clinical trials.
Which Antiphospholipid Antibodies Should Be Measured in the Antiphospholipid Syndrome?
Authors: Galli Monica
Published: April 2003

The results of an analysis of studies published over a 10-year period that addressed the association between antiphospholipid antibodies (aPLs) and thromboembolic events in patients with the antiphospholipid syndrome (APS) are reported. It would appear that lupus anticoagulants are the strongest risk factor for thromboembolic events in aPL-positive patients. Consequently, it is suggested that all patients with clinical manifestations of APS should be tested for these antibodies. On the other hand, the results do not unequivocally show that measurement of anticardiolipin antibodies is of help in defining the thrombotic risk of patients. Furthermore, they only partially support the notion that anti-β2-glycoprotein I and antiprothrombin antibodies may be independent risk factors for thrombosis.
Death after Joint Replacement
Authors: Frostick Simon P.
Published: April 2003

This review considers the incidence of fatal pulmonary embolism following lower limb arthroplasty and femur neck fracture and considers whether some of these deaths are preventable. The data considered refer to short-term in-hospital prophylaxis with low-molecular-weight heparin and unfractionated heparin. Mortality rates depend on the type of surgery, with fracture of the femur being associated with the highest rates and knee replacement associated with the lowest. Short-term prophylaxis with dalteparin does appear to reduce the risk of death, but the risk extends well beyond discharge from hospital. Studies of extended out-of-hospital prophylaxis in order to reduce deaths related to thromboembolism are warranted.
Low-Molecular-Weight Heparins: Are They Interchangeable?
Authors: van der Heijden Jeroen F. , Prins Martin H. , Büller Harry R.
Published: April 2003

Unfractionated heparin (UFH) has been used as an antithrombotic agent in the treatment of various clinical entities for over 60 years. Low-molecular-weight heparin (LMWH) com- pounds have gradually replaced UFH for these indications as they have several advantages, including subcutaneous administration and the lack of need for laboratory monitoring. Ever since their introduction, there has been discussion about whether LMWH compounds differ in their efficacy and safety. The best answer is given by direct comparison of two or more preparations; however, such trials are very scarce. Comparison using classical meta-analysis is limited as only a small number of trials with the respective low-molecular-weight heparin compounds are available. The objective of the present analysis has been to use a novel way of plotting the odds ratios of the different studies to compare the efficacy and safety of different LMWH compounds in the initial treatment of patients with venous thromboembolism. Classical meta-analysis revealed reductions in safety and efficacy of 30–40% in favour of LMWHs. Contrasting the log odds ratios of efficacy and safety indicated that there is no conclusive evidence that LMWHs have intrinsic different safety and efficacy profiles.
IL–12–Dependent, IFN–γ–Independent Experimental Glomerulonephritis
Authors: Le Hir Michel , Ryffel Bernhard , Schatzmann Ursula
Published: April 2003

There is evidence that crescentic glomerulonephritis initiated in rodents by heterologous antibodies against the glomerular basement membrane (anti–GBM glomerulonephritis) depends on a Th1–type immune reaction. Interleukin 12 (IL–12) is crucial for the development of Th1 helper cells, and interferon gamma (IFN–γ) is a major proinflammatory product of these cells. In order to test the role of the two cytokines in anti–GBM glomerulonephritis we used mice lacking either the p40 chain of IL–12 (IL–12–/–) or the IFN–γ receptor (IFN–γR–/–). Glomerulonephritis was induced by injecting a rabbit anti–GBM serum in mice preimmunized against rabbit IgG. Glomerulonephritis was assessed on the basis of proteinuria, immunofluorescence findings and histology. IL–12–/– mice were completely protected against glomerulonephritis. In contrast, IFN–γR–/– mice were more severely affected than wild–type mice. Similarly, cutaneous delayed–type hypersensitivity, a typical Th1 response, was abolished in the IL–12–/–, mice but increased in the IFN–γR–/– mice. The data obtained in IL–12–/– mice support the view that crescentic glomerulonephritis in this model represents a Th1 response. Since IFN–γ is not required, other products of Th1 cells are likely to mediate glomerulonephritis.
Dilazep Hydrochloride, an Antiplatelet Drug, Inhibits Lipopolysaccharide-Induced Mouse Mesangial Cell IL-6 Secretion and Proliferation
Authors: Gohda Tomohito , Makita Yuichiro , Shike Toshihide , Funabiki Kazuhiko , Shirato Isao , Tomino Yasuhiko
Published: April 2003

Background: Antiplatelet agents have been widely used to reduce proteinuria and to prevent the progression of chronic glomerulonephritis or diabetic nephropathy to end-stage renal failure. Dipyridamole, one type of antiplatelet drug, inhibits the proliferation of glomerular mesangial cells (MCs). The effect of dilazep hydrochloride (dilazep) on these cells is still obscure. The effects of dilazep on cultured MC IL-6 secretion and proliferation were investigated in the present study. Methods: IL-6 secretion from MC induced by bacterial lipopolysaccharide (LPS) were assessed using sandwich ELISA. LPS-induced MC proliferation was detected by 3H-thymidine incorporation and WST-1 assay (similar to MTT assay). Results: Incubation of MCs with various dosages of LPS (0, 1, 10, 50 and 100 ng/ml) induced IL-6 secretion in a dose-dependent manner. However, dilazep significantly inhibited this LPS-induced IL-6 secretion from MCs in a dose- and time-dependent manner. Dilazep also significantly inhibited MC proliferation in a dose-dependent manner. Conclusion: It appears that these effects of dilazep may prevent progression of mesangial proliferative glomerulonephritis.