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Inadvertent immunisation of seronegative women with RA27/3 rubella virus live-attenuated vaccine several weeks before and after conception is described. Whereas in 5 cases the vaccine virus was not transmitted vertically, in 1 case vaccination led to the development of persistent fetal infection with prolonged virus shedding for more than 8 months. Sequence analysis carried out on isolates from amniotic fluid, from cord blood leukocytes as well as from infantile urine confirmed an infection by the vaccine strain. At birth, the newborn infant exhibited none of the symptoms compatible with the congenital rubella syndrome and signs indicative for development of late onset disease are not apparent. This observation constitutes the first unequivocal documented case of rubella vaccine virus related to persistent fetal infection.

Ketamine is a drug widely used for analgesia and sedation of children for diagnostic and therapeutic procedures. The authors investigated in a randomized controlled clinical trial if diazepam premedication would have a beneficial effect on side effects related to ketamine anesthesia for bone marrow punctures (BMPs) in children with acute lymphoblastic leukemia (ALL). Sixteen children 4 years or older at the time of BMP were eligible. The first 2 BMPs after complete remission was obtained were studied. BMPs were performed under ketamine anesthesia (1.0-1.5 mg/kg i.v.), as usual. Patients were randomized to receive 1 h before the first BMP blinded, either diazepam or placebo orally and before the second BMP the other way round. Blood pressure, heart rate, and oxygen saturation were monitored, and patients were observed for signs of anxiety, pain, and other side effects. The patients were interviewed after each BMP and asked for their preference 1 week after the second BMP. Ketamine anesthesia appeared as safe and effective after diazepam premedication as after placebo premedication. From the interviews and questionnaires, it was clear that half of the children preferred diazepam premedication because of less awful dreaming and more gradual falling asleep and waking up. Diazepam premedication may be useful for selected children with ALL receiving ketamine anesthesia for BMPs.

The low density lipoprotein receptor plays an important role in the uptake of maternal plasma lipoproteins for placental steroid metabolism. Using an RNase protection assay, the expression of low density lipoprotein receptor mRNA was measured in human placentas from pregnancies in the second and third trimester and from pregnancies with intrauterine growth retardation. Pregnancies complicated by intrauterine growth retardation showed a significantly higher expression (1.28), compared with the term and preterm pregnancies (P0.05). The increased low density lipoprotein receptor expression in placentas from pregnancies with intrauterine growth retardation could be a compensatory mechanism for the lower circulating low density lipoprotein concentrations in women with intrauterine growth retardation.

We report two cases of premature rupture of membranes and oligohydramnios (gestational age: 23rd/24th week). In both pregnancies artificial instillation of amniotic fluid (AIF) was performed once a week. We hypothesize that fetal head compression with impaired cerebral perfusion due to oligohydramnios causes periventricular leukomalacia. Prolongation of gestation of 36/23 days without neurological lesion support this method. Surveillance of these pregnancies in a perinatal center has to include very early detection of chorioamnionitis. If not so, gained prolongation of pregnancy with a lower rate of cerebral damage becomes harmful due to infection. Clinical benefits and disadvantages of this technique have to be evaluated in a prospective randomized trial.

We describe a giant bilateral vascular mass in the skin of the chest diagnosed by ultrasound investigation in a fetus of 20 gestational weeks. Ultrasound and colour Doppler investigations detected no signs of early congestive heart failure but rapid and excessive enlargement of the vascular mass. Indications of increased fetal blood volume were found. The fetus had additional minor anomalies. Early in utero manifestation suggests a severe vascular malformation.

A 28-year-old woman was referred at 33 weeks of gestation with suspected fetal intestinal atresia. Sonography showed a large extra-abdominal mass on the right of the normal umbilical cord insertion. Following Cesarean section at 36 weeks and immediate surgical treatment, the malformation was not definable either as an omphalocele or as gastroschisis. This reported case involves a previously undocumented malformation of the fetal abdominal wall described as a 'hernia' of the fetal abdominal wall.

In two Dutch subjects who had been on holiday in the tropics, a woman aged 32 and a man of Surinam descent aged 52 years, and in two men aged 21 and 38 years who had arrived from the tropics in the Netherlands, one recently and one 15 years previously, import skin diseases were diagnosed: larva migrans cutanea, cutaneous leishmaniasis, mycetoma and lobomycosis. The diagnosis was based on the anamnesis, the clinical picture and histopathological findings. The patients were cured by administration of antimicrobial agents and (or) excision. When travellers or immigrants from the tropics present with skin lesions, an imported skin disease should be considered.

C-type natriuretic peptide (CNP) is the third member of the natriuretic peptide family that is involved in the regulation of blood pressure, renal function and volume homeostasis. In man CNP is highly expressed in endothelial cells, distinct brain areas and kidney. Maternal CNP plasma concentration remains stable in normal pregnancy with advancing gestational age. Using a radio-immunoassay we measured for the first time CNP plasma levels of women in the active phase of labour, during elective cesarean section in spinal anesthesia and in the postpartum period. Healthy pregnant women in the third trimenon were used as controls (7.36 +/- 3.0 pg/ml). CNP is significantly increased in labour (11.48 +/- 1.2 pg/ml) as well as after volume load before cesarean section (14.25 +/- 4.5 pg/ml). The postpartum concentration (9.3 +/- 2.4 pg/ml) was in the range of the controls. We conclude that CNP in plasma is increased during spontaneous labour. Its possible role in volume regulation with increased levels after rapid volume load but normal levels in puerperium with physiologically enhanced diuresis and natriuresis remains unclear.

Objective: To measure plasma levels of C-type natriuretic peptide in women with normal pregnancies and those complicated by gestational hypertension and preeclampsia. Methods: We collected venous plasma samples from 20 women with normal pregnancies, 15 with gestational hypertension, and nine with preeclampsia. Gestational ages ranged from 26–39 weeks. We measured C-type natriuretic peptide concentrations by radioimmunoassay. Statistical comparisons were made by one-way analysis of variance and Tukey test. Results: The mean plasma concentration of C-type natriuretic peptide in the women with normal pregnancy was 7.1 ± 2.5 pg/mL, in those with gestational hypertension 9.6 ± 4.2 pg/mL, and in those with preeclampsia 8.1 ± 2.4 pg/mL. There were no statistically significant differences between groups. The statistical power to detect a difference of 3 pg/mL at the 5% significance level was 78% for the group with gestational hypertension and 64% for the preeclamptic group. Conclusion: Our results suggest that C-type natriuretic peptide plasma levels are not significantly different in normal pregnancies and those complicated by gestational hypertension or preeclampsia. In hypertensive pregnancy disorders, C-type natriuretic peptide showed a distinct course compared with atrial natriuretic peptide and brain natriuretic peptide.

A prematurely born infant developed rectal blood loss several hours after birth, after his first formula feeding. Discontinuing the feeding resolved symptoms, but after resuming feeding rectal blood loss reappeared. There were no signs of necrotizing enterocolitis. Suspecting cow's milk allergy, the feeding was changed to a casein-based protein hydrolysate, without effect. Meanwhile, laboratory tests indicated cow's milk allergy. Symptoms only resolved after introducing an amino acid-based formula supporting a definite diagnosis of cow's milk-induced allergic colitis. This is the first description of a premature infant with symptoms of allergic colitis, appearing within hours after birth, suggestive of intrauterine sensitization. The exact mechanisms of sensitization remain obscure. CONCLUSION: Cow's milk-induced allergic colitis can occur after the first feed, even in a prematurely born neonate. This is most probably due to intrauterine sensitization, and should be included in the differential diagnosis of rectal blood loss.

Vaginal colonization with Mycoplasma hominis and Ureaplasma urealyticum has been implicated as a cause of prematurity. Several mechanisms to induce preterm labor have been discussed. The investigation of expressed and secreted enzymes requires a feasible method for culturing and further processing of these bacteria. We describe a simple technique for culturing of Mycoplasma hominis and Ureaplasma urealyticum without contamination with other microorganisms and isolating DNA and supernatant. PCR amplification of a chromosomal Mycoplasma fragment was performed as positive control.

C-type natriuretic peptide (CNP) is a recently identified member of the family of natriuretic peptides that plays an important role in the regulation of blood pressure, renal function and volume homeostasis. Its effects are mainly hypotensive and natriuretic. CNP is also considered to be an autocrine/paracrine regulator of the endothelium as well as a neuropeptide. Little is known about its role as a circulating substance. There are only few data of CNP plasma levels and changes in disease. Using a highly specific radioimmunoassay we established for the first time CNP plasma levels of normal pregnancies 5.9 +/- 1.5-8.6 +/- 2.1 pg/ml) and a non-pregnant control group (8.1 +/- 1.7 pg/ml). In contrast to atrial natriuretic peptide and brain natriuretic peptide we have not found significant differences of the plasma levels during normal pregnancy and no difference to the non-pregnant control group.

In an earlier study, we generated a large number of Mycobacterium leprae-responsive and M. leprae-nonresponsive T cell clones (TCC) from the lesional skin of immunologic unstable borderline leprosy patients. In that study, we divided TCC into type 1- and type 2-like on the basis of their IFN-gamma and IL-4 expression. To explore whether other cytokines are coproduced along with IFN-gamma and IL-4, we investigated the secretion of a panel of other cytokines (TNF-alpha, IL-5, IL-6, IL-10, and IL-13) by a large number of these TCC. Upon analysis of 139 M. leprae-responsive TCC, we observed a positive correlation in the coproduction of IFN-gamma/TNF-alpha (r = 0.81), and in that of IL-4/IL-5 (r = 0.83), IL-4/IL-13 (r = 0.80), and IL-5/IL-13 (r = 0.82). Polarized type 1-like TCC produced dominantly IFN-gamma/TNF-alpha, and polarized type 2-like TCC predominantly IL-4/IL-5/IL-13. Most type 0-like TCC produced both sets of cytokines. In contrast, type 1- and type 2-like subsets of M. leprae-nonresponsive TCC (n = 58) did not show the same coexpression of these cytokines. Furthermore, when the differential expression of a broad panel of cytokines by individual M. leprae-responsive TCC is considered, it appeared that additional phenotypes could be recognized. These results suggested that distinct isotypes of type 1- and type 2-like T cells, based on the secretion of a panel of cytokines, may reflect M. leprae-specific characteristics.

The study aims to test the influence of vaccination with polyvalent vaccine prototypes on the vibriosis resistance of Penaeus monodon. Vaccinated P. monodon post-larvae were challenged with a virulent Vibrio alginolyticus strain 10, 20, and 30 days post-vaccination. Results showed that vaccination significantly enhances the resistance of shrimp to vibriosis. This effect decreased by age of the post-larvae, particularly for the glucan group.

Borderline leprosy patients often undergo acute changes in immune reactivity that manifest as reversal reaction (RR) in the course of the disease. RR is associated with an exacerbated local delayed-type cellular immune response to Mycobacterium leprae and is responsible for severe tissue damage. We investigated whether RR episodes are associated with a change in T cell subsets in the lesional skin with regard to their cytokine secretion profiles. M. leprae-responsive T cell lines and thereafter T cell clones (TCC) were generated from the lesional skin of seven untreated borderline leprosy patients (with or without RR) and again from three of these patients experiencing RR during treatment. The phenotypes of the M. leprae-responsive TCC were either CD4+, CD8+, CD4-/CD8+/TCR gammadelta+, or CD4-/CD8-/TCR gammadelta+, although most of them were CD4+. Regardless of the clinical status of the untreated patients, a major subset of the M. leprae-responsive TCC was type 0-like and produced both IFN-gamma and IL-4. Interestingly, in all three patients who experienced a (re)occurrence of RR during treatment after the first analysis, a clear shift to polarized IFN-gamma production by the M. leprae-responsive TCC (type 1-like) was observed. This shift in T cell subsets was also reflected in the observed decrease in serum IgG and IgM levels of the same patients during RR. These finding indicate that CD4+ M. leprae-responsive T cells with a polarized type 1-like phenotype might be responsible for the immune-mediated tissue damage occurring during RR.

IL-12 secretion by APC is critical for the development of protective Th1-type responses in mycobacterial (Mycobacterium avium and Mycobacterium tuberculosis) infections in mice. We have studied the role of IL-12 and IL-2 in the generation of Mycobacterium leprae-specific T cell responses in humans. Leprosy patients were defined as low/nonresponders or high responders based on the level of T cell proliferation in M. leprae-stimulated PBMC. In high responders, M. leprae-induced proliferation was markedly suppressed by neutralizing anti-IL-12 mAb (inhibition 55 +/- 6%). Neutralization of IL-2 activity resulted in an inhibition of 77 +/- 4%. Given the importance of endogenous IL-2 and IL-12 in M. leprae-induced responses, we investigated the ability of rIL-2 and rIL-12 to reverse T cell unresponsiveness in low/nonresponder patients. Interestingly, rIL-12 and rIL-2 strongly synergized in restoring both M. leprae-specific T cell proliferation and IFN-gamma secretion almost completely to the level of responder patients. A similar synergy between rIL-2 and rIL-12 was also observed in high responders when suboptimal M. leprae concentrations were used for T cell stimulation. Our data demonstrate a crucial role for endogenous IL-12 and IL-2 in M. leprae-induced T cell activation. Most importantly, we show that rIL-2 and rIL-12 act in synergy to overcome Ag-specific Th1 cell unresponsiveness. These findings may be applicable to the design of antimicrobial and antitumor vaccines.

This paper focuses on the intersection point between the strategic and operational levels of war. It recommends that aerial coercion become a formal part of Joint Doctrine because (1) Engagement and Enlargement created new military requirements that remain uncovered, (2) there already exist well-developed methods of aerial coercion available for use, and (3) the problems that bedeviled aerial coercion are no longer insurmountable. This paper focuses on the intersection point between the strategic and operational levels of war. It recommends that aerial coercion become a formal part of Joint Doctrine because (1) Engagement and Enlargement created new military requirements that remain uncovered, (2) there already are well-developed methods of aerial coercion available for doctrinal use, and (3) the problems that bedeviled aerial coercion in the past are no longer insurmountable.

To investigate the influence of recommendations on the quality of postoperative pain management in children.Prospective.University Hospital Groningen, the Netherlands.After interdisciplinary recommendations on postoperative pain were developed, the quality of postoperative pain management was investigated before implementation (phase I; n = 50 children aged 0-14 who underwent elective surgery), three months after the implementation (phase II; n = 51), and nine months later (phase III; n = 50). Quality was defined by a pain score (for ages 0 to 4 with the 'Children's Hospital of Eastern Ontario pain scale' (CHEOPS) and for ages 4-14 with the Oucher scale) and the prescription of analgetics: kind, dose, frequency, prescription by anaesthetist and doctor on the ward. Pain was scored every 2 hours during the first 24 hours after surgery. A CHEOPS scoreor = 6 an Oucher scoreor = 50 was defined as adequate; higher scores were defined as inadequate.Pain measurement showed a statistically significant improvement of pain scores in time (phase II and III compared with phase I: odds ratio: 2.5; 95% confidence interval: 1.03-6.00; p0.01). Searching for factors that could be responsible for this improvement, like medication, we found no statistically significant differences in everyday practice in phase II and III compared with phase I. However, children who could score their pain by self-report (Oucher) showed the best results in all 3 phases of the study. The youngest children, i.e. less than 6 months old, showed inadequate results during the whole study. The greatest improvement in time during the first 12 hours was seen in the group of children older than 6 months. The recommendations were followed more strictly in younger children, and when continuous morphine was given.Pain scores in children improved after the introduction of recommendations on postoperative pain. However, the improvement could not be attributed to factors like medication. Factors like a change in attitude towards pain could be responsible for this change.

The pathogenesis of venous leg ulcers is still not known. One hypothesis states that pericapillary fibrin cuffs might play an important role. These fibrin cuffs might not be broken down because of an inadequate fibrinolytic activity.To stimulate fibrinolysis, tissue type plasminogen activator (t-PA) containing ointment was applied over 12 weeks on chronic venous leg ulcers of six patients. Three of the six ulcers healed within this period. Biopsy specimens for immunofluorescence studies were taken from the ulcer base and margin before and at the end of treatment and, if the ulcer had healed, from the healed area.Deposition of pericapillary fibrin was seen around the capillaries of all investigated ulcers at the start of the study. Pericapillary fibrin was still present with nearly undiminished intensity at the end of the study even though 3 of the ulcers healed.It is most likely that the healing of these ulcers was not improved by the fibrinolytic activity of t-PA, but caused by other wound healing properties of t-PA.

This report argues that current members of U.S. aerospace forces do not have a language or lexicon to differentiate one theory of airpower from another. To solve this problem, this report (1) describes the prison house of language that thwarted airpower theorists in the past, (2) provides a model to analyze airpower theory properly, and (3) reviews different theories of airpower, with a primary emphasis on five post 1960 theorists.

Using a discriminance analysis we investigated the interactions between parameters of the vaginal flora and subclinical infection regarding preterm birth. The prospective study includes 222 single pregnancies, 114 of them with preterm labour or premature rupture of the membranes (PROM). The analysis includes information of a vaginal/ cervical smear (total number of pathogenic germs, species and groups of pathogenic germs), vaginal pH, maternal white blood cell count (WBC), C-reactive protein (CRP) and temperature. A normal vaginal flora was found in only 19%. Pregnancies with preterm labour have a higher quantity of pathogenic germs, whereas there is no difference of the germ group distribution. Moreover, pregnancies with preterm labour and 2 or 3 pathogenic germs have a higher rate of preterm birth. The discriminance analysis shows that only the affiliation the risk group with preterm labour and the parameters CRP and WBC have a significant selectivity for consequent preterm birth, but not the parameters containing information of the vaginal flora. The same is valid for the group with preterm labour, where only the parameters PROM and WBC are able to select significantly for preterm birth.

The faulty origin of the left coronary artery from the pulmonary artery is with an incidence of 1:300,000 newborns a very rare heart defect. We report a case of a pregnancy with two intrauterine blood transfusions in the 30th and 32nd week of gestation because of Rh-incompatibility and fetal anaemia. Dopplersonographic and echocardiographic parameters were normal. In the 32nd week of gestation delivery was induced (birth weight 2240 g, cord pH value 7.35, Apgar-score 8/9/9). Under a therapy with respiration, blood exchange transfusion and cardiotonic drugs the newborn died in the second week. The autopsy showed a general immaturity, a haemosiderosis of spleen, liver and lungs, a marked cellular jaundice and signs of a multi-organ-failure. The sinus of the pulmonary valve was the origin of the left coronary artery. In case of a seriously impaired pumping action and after exclusion of other heart defects the Bland-White-Garland-Syndrome (BWGS) has to be considered. Although a prenatal diagnosis of BWGS is with high resolution-ultrasound possible, the early postnatal diagnosis seems to be more relevant.

Ascending infections of the female genital tract as a cause of preterm birth have become major importance regarding the prevention of preterm birth. Measurements of the vaginal pH value are able to verify a alkalinisation of the vagina caused by a atypical vaginal flora. In a prospective study we measured the vaginal pH value using indicator paper and evaluated the results of a vaginal and cervical smear of 162 single pregnancies from 26 to 30 weeks of gestation. In contrast to normal pregnancies there is a relation between a pathological pH value4.5 and consequent preterm birth in pregnancies with preterm labour but also in pregnancies with other risk factors without symptoms of preterm labour.

We investigated 69 healthy women with normal pregnancy and delivery in order to establish normal values for the second trimester of gestation. In a cross-sectional study we measured the pulsatility index (PI), the maximal systolic velocity (V-MAX) and the mean velocity (V-MEAN) in the uterine arteries (UAA), the umbilical artery (UA), the fetal thoracic aorta (FTA) and the middle cerebral artery (MCA) from the 14th to the 24th week of gestation. A regression analysis shows a significant slope of the PI of UAA and UA (p0.01), whereas in contrast to the UA there is no significant difference of the median values of the UAA-PI from the 18th week of gestation. With progressive gestation V-MEAN of UAA increases more (p0.001) than V-MAX (p0.05) as a result of the increasing diastolic blood flow. In the UA there is also a significant rising trend of V-MAX and V-MEAN (p0.0001). No significant changes of the PI of FTA and MCA are seen, although there is a slight increasing trend. After the 20th week in the MCA a marked tendency to high PI-values is observed, whereas there is a change between different flowpatterns prior to the 20th week. V-MAX and V-MEAN of FTA increases significantly, but there is no change in the MCA. Our results for UAA and UA correspond to the known morphologic changes in the utero- and fetoplacental vessels in the second trimester and are comparable to other results. Particularly the physiology of the cerebral perfusion needs to be investigated more intensively. These normal values provide a good basis for early and complex assessment of a disturbed perfusion in the second trimester.

The three-dimensional structures of two cytochromes c′ have been determined in order to analyse the common features of proteins of this family and their relationship with other four-helix bundle structures. The structure of cytochrome c′ from Alcaligenes sp was determined by molecular replacement supplemented with the iron anomalous scattering and the use of a single isomorphous heavy-atom derivative, and was refined using synchrotron data to 1.8 A resolution. The final model, comprising 956 protein atoms (one monomer) and 89 water molecules, has a final R value of 0.188 for all data in the range 20.0–1.8 A resolution (14 673 reflections). The structure of the cytochrome c′ from Alcaligenes denitrificans is isomorphous and essentially identical (r.m.s. deviation for all atoms 0.36 A). Although its amino-acid sequence has not been determined chemically, only four differences from that of Alcaligenes sp cytochrome c′ were identified by the X-ray analysis. The final model for Alcaligenes denitrificans cytochrome c', comprising 953 protein atoms and 75 water molecules, gave a final R factor of 0.167 for all data in the range 20.0–2.15 A (8220 reflections). The cytochrome c′ monomer forms a classic four-helix bundle, determined by the packing of hydrophobic side chains around the enclosed haem group. There are very few cross-linking hydrogen bonds between the helices, the principal side-chain hydrogen bonding involving one of the haem propionates and a conserved Arg residue. The cytochrome c′ dimer is created by a crystallographic twofold axis. Monomer–monomer contacts primarily involve the two A helices, with size complementarity of side chains in a central solvent-excluded portion of the interface and hydrogen bonding at the periphery. Both species have a pyroglutamic acid N-terminal residue. The haem iron is five-coordinate, 0.32 A out of the haem plane towards the fifth ligand, His120. The unusual magnetic properties of the Fe atom may be linked to a conserved basic residue, Arg124, adjacent to His120.

To study the usefulness of haemolymph for shrimp health assessment characteristics of both the cellular and humoral composition of growing juvenile Penaeus monodon (n = 16) were determined. Haemolymph was obtained by a puncture of the ventral part of the haemocoel. Using routine staining methods five different cell types could be distinguished by light microscopy. Electron microscopy studies revealed granulocytes, semigranulocytes and hyalinocytes. Mean total haemocyte count was 50.9 × 106 cells/ml whereas mean haemolymph plasma protein content was 79.9 g/l consisting of one major and one or two minor fractions.

We examined 114 women with preterm labor using discriminance analysis to assess predictive values in terms of prolongation of gestational age and gestational age at the time of delivery. We used parameters like medical history, clinical features, infections, Doppler sonography and cardiotocography. Maternal temperature, cardiotocographic findings, premature rupture of membranes, number of abortions and pregnancy terminations and cervical dilatation at the time of admission contribute significantly to predict prolongation of pregnancy (or = as well as7 days). Our results from this analysis showed sensitivity and specificity of 0.70 and 0.97 and positive and negative predictive values of 0.89 and 0.91, respectively. We observed significant differences concerning parameters like preterm rupture of membranes, cervical dilatation, pathological bacteria in the vagina and pathological Doppler values between subsequent preterm and term deliveries in pregnancies with a prolongation of7 days was. Upon reclassifying our results, we obtained sensitivity and specificity of 0.69 and 0.89 and positive and negative predictive values of 0.78 and 0.84, respectively. Early prognostic assessment of preterm labor can be made with discriminance analysis. The results inspire us to perform a prospective examination of the therapy plan that we have obtained.

In a prospective study we investigated 100 twin pregnancies in order to assess the predictive value of doppler-sonographic measurements regarding the pregnancy outcome. We used the pulsatility index of the uterine arteries, the umbilical artery, the fetal aorta thoracica and the fetal middle cerebral artery. The measurements were obtained at various gestational ages between 24 and 32 weeks. Significant differences between unselected twin pregnancies and normal single pregnancies were not found. Moreover, there is no relation between the severity of an impaired perfusion and an adverse outcome of the pregnancy. The prediction of fetal distress (hypoxia, preterm birth, poor fetal growth, acidosis, and disturbed neonatal adaption) with doppler-score has a sensitivity of 25% and a positive predictive value of 63%. From this rather insufficient result we conclude that doppler screening in unselected twin pregnancies hardly proves to be useful.

Background: Obesity is associated with a prothrombotic state, which may contribute to the increased risk of thrombotic events. Objective: To assess the effects of (pre)adipocyte-derived adipokines on fibrinogen, plasminogen activator inhibitor-1 (PAI-1) and tissue factor (TF) production by hepatocytes. Methods: HepG2 hepatocytes were incubated with conditioned media (CM) derived from preadipocytes and adipocytes, which had been untreated or prestimulated with tumor necrosis factor (TNF)-α, interleukin (IL)-1β or IL-6. After 24 h, supernatants and cell lysates were harvested for measurement of fibrinogen, PAI-1 and TF. Results: (Pre)adipocyte CM significantly enhanced the production of PAI-1 by HepG2 cells 2.5- to 4.4-fold. CM from cytokine-stimulated (pre)adipocytes significantly induced fibrinogen secretion 1.5- to 4.2-fold. TF production was not affected by the CM. After specific depletion of TNF-α, IL-1β or IL-6 from the CM, IL-6 was shown to be the most prominent stimulus of fibrinogen secretion and IL-1β of PAI-1 secretion. In addition, fibrinogen, PAI-1 and tissue factor production was evaluated by direct stimulation of HepG2 cells with TNF-α, IL-1β or IL-6. IL-6 enhanced fibrinogen synthesis 4.3-fold (P

To inventory the changes in leprosy epidemiology in the Netherlands.Retrospective.Academic Medical Centre (Amsterdam) and University Hospital Dijkzigt (Rotterdam), the Netherlands.The medical records of all new leprosy patients in the period 1970-1991 were analysed.Between 1970 and 1991, 622 new leprosy patients were registered; 371 men (59.6%) and 251 women (40.4%). Most patients came from Surinam (73.3%) and Indonesia (7.2%). The mean time lapse between onset and treatment in the Netherlands was 10.1 years. Switching from monotherapy to combination therapy (1979) had no effect on the incidence of reversal reactions (cellular hypersensitivity in immunologically unstable patients), but did affect the incidence of erythema nodosum leprosum during the treatment.Leprosy in the Netherlands is an important disease, mainly from Surinam. The main advantage of combination therapy is the shortened duration of treatment. The treatment of choice is the one recommended by the WHO, the combination therapy with rifampicin administration once a month, because of the few adverse effects.

A pregnancy with a partial or complete hydatidiform mole and a coexistent living fetus is a rare condition. Most cases are consistent with a twin pregnancy, with distinct borders between placenta and mole on sonography. We report a case with a single placenta with cystic structures resembling histologically a complete hydatidiform mole. The pregnancy resulted in a diploid female fetus in the 34th week of gestation with marked hypotrophy. The non-molar part of the placenta showed a complex alteration of villous maturity resulting in intrauterine growth retardation but a living fetus. No malignancy occurred post partum. The clinical and pathogenetic aspects of this rare entity are discussed.

The aim of the present study was determination of the complex character of hemodynamic disturbances in uteroplacental, fetoplacental and fetal vessels as well as their influence on the outcome of pregnancies suspected to have IUGR on ultrasound examination. A special form of Doppler scoring was implemented for documentation of the hemodynamic disturbances. Of 82 single pregnancies with sonographically suspected IUGR, 30 women gave birth to eutrophic babies whereas 52 mothers delivered hypotrophic newborns. However, 43 cases of the hypotrophic babies had birth weight below the 5th percentile. The median values of Doppler indices for those with eutrophic babies revealed no significant differences compared to normal collective. However, significantly elevated RI and PI values in both uterine arteries, the umbilical artery and fetal descending aorta of pregnancies which resulted in hypotrophic babies indicated hemodynamic disturbance. The umbilicoplacental and uteroplacental vasculature compartments were almost equal involved with a proportion of 39% or 30% respectively. Birth weightor = 5th percentile was associated with a further increase of pathological values in this both vascular areas. Significant rang correlations between values of Doppler indices and clinical parameters indicated a direct association between perfusion disturbances and fetal outcome of the pregnancies studied. The present Doppler score, which is a summary of all separately evaluated examined blood vessels, is suitable for clinical implementation and showed a good correlation to the clinical parameters in our study. Therefore Doppler perfusion measurements should play an important role in the clarification of suspected fetal growth retardation.

Moles and coexisting living fetus are rare. Most cases are twin gestations. We report a case of a single placenta with focal hydatidiform mole of the placenta and a coexisting living fetus. The fetus with hypotrophy was born in the 34th weeks of gestation with a normal diploid female karyotype. The placenta showed a complex alteration of the non molar part with consecutive vascular hyperplasia. The differential diagnosis of molar pregnancies and coexistent living fetuses are discussed. Postpartal are cytogenetic examination of fetal tissue (skin biopsy) and both parts of the placenta, the molar and non molar, necessary. In each case should be a postpartal measurement of beta-HCG in serum of the mother to rule out a gestational trophoblastic disease.

Background: Venous hypertension of the hand is an unusual complication of an arteriovenous shunt in patients receiving dialysis. Objective: We investigated whether in venous hypertension of the hand pericapillary cuffs are present and whether they consist of fibrin and other components. Methods: Biopsy specimens were taken from three patients with a side-to-side shunt and venous hypertension of the hand, from three patients with an end-to-side shunt without venous hypertension, and from three control subjects. The specimens were stained with antibodies against fibrinogen, fibrin, plasminogen activator inhibitor-I, factor VIII-related antigen, and collagen type IV. Results: Pericapillary cuffs consisting of fibrin only were observed in patients with venous hypertension of the hand. In this fibrin network, factor VIII-related antigen and collagen type IV were also present. The finding of plasminogen activator inhibitor-I in the pericapillary cuff in the patient with the most severe signs of venous hypertension may indicate that breakdown of this fibrin cuff is impaired. Conclusion: Pericapillary cuffs may be of pathogenetic significance in venous hypertension of the hand.

The granulomatous skin lesions in leprosy are thought to be initiated by the immune response to certain antigens of the causative agent, Mycobacterium leprae. The antigen 85 complex is one of the major targets in the immune response to M. leprae infection. In the present study, a panel of previously characterized monoclonal antibodies (MAbs) (3A8, Rb2, A4g4, A2h11, Pe12, and A3c12) reacting with different epitopes of the 85 complex proteins of Mycobacterium tuberculosis and M. leprae was employed in a comparative immunohistological analysis to demonstrate the in situ expression of 85 complex antigenic epitopes in leprosy lesions across the clinical spectrum and in M. leprae-infected armadillo liver tissues. These MAbs showed a heterogeneous staining pattern in a given leprosy lesion. In highly bacilliferous borderline and lepromatous leprosy lesions, MAbs Rb2, A4g4, A2h11, and Pe12 stained clear rod-shaped M. leprae bacilli within macrophages, and the degree of staining correlated with the bacillary index of the lesion. On the other hand, MAbs 3A8 and A3c12 staining was mostly seen as a diffuse staining pattern within interstitial spaces and on the membranes of the infiltrated cells but not the bacilli. In paucibacillary borderline and tuberculoid leprosy lesions, only 3A8, Rb2, and A3c12 showed distinct staining in association with infiltrates in the granuloma. None of these MAbs showed any detectable reaction with control nonleprosy skin lesions, while MAb A3c12 positively stained the granulomas of both leprosy and control specimens. In situ reactivity of these MAbs with M. leprae-infected armadillo liver tissues also showed a heterogeneous staining pattern. Interestingly, a clear difference in expression of these epitopes was observed between armadillo tissues and human leprosy lesions. By immunogold ultracytochemistry, we further showed the differential localization of these MAb-reactive epitopes on the cell surface, in the cytosol, and at the vicinity of M. leprae within Kupffer cells of armadillo liver tissues. Our results indicate that these antigenic epitopes of the antigen 85 complex are differentially expressed in leprosy lesions and infected armadillo tissues and that they could be target determinants in the immunopathological responses during M. leprae infection.

In recent years the knowledge of pain in pediatric patients has grown. Attitudinal changes and the development of assessment-scales also contributed to a better understanding of childhood pain. This article provides a guidance for the pharmacological management of pain in children.

The phosphocarrier protein IIIGlc is an integral component of the bacterial phosphotransferase (PTS) system. Unphosphorylated IIIGlc inhibits non-PTS carbohydrate transport systems by binding to diverse target proteins. The crystal structure at 2.6 A resolution of one of the targets, glycerol kinase (GK), in complex with unphosphorylated IIIGlc, glycerol, and adenosine diphosphate was determined. GK contains a region that is topologically identical to the adenosine triphosphate binding domains of hexokinase, the 70-kD heat shock cognate, and actin. IIIGlc binds far from the catalytic site of GK, indicating that long-range conformational changes mediate the inhibition of GK by IIIGlc. GK and IIIGlc are bound by hydrophobic and electrostatic interactions, with only one hydrogen bond involving an uncharged group. The phosphorylation site of IIIGlc, His90, is buried in a hydrophobic environment formed by the active site region of IIIGlc and a 3(10) helix of GK, suggesting that phosphorylation prevents IIIGlc binding to GK by directly disrupting protein-protein interactions.

The aim this study to compare perfusion parameters, measured by Doppler sonography, between normal pregnancies and pregnancies with symptoms of preterm labor. The patient collective comprised of 3 groups: 62 normal pregnancies (group 1), 23 preterm labor before tocolysis (group 2) and 114 preterm labor up to 48 hours under tocolysis (group 3). In each case pulsatility index was measured in the uterine artery on the placental side and non-placental side, umbilical artery, fetal descending aorta and middle cerebral artery. In the contrary to patients in group 1, patients in group 2 and 3 showed significantly elevated PI values in uterine artery placental side and non-placental side, respectively. The median of PI for both uterine arteries in both groups was significantly higher compared to that of group 1. However, 1/3 of patients with preterm labor showed pathologic uterine perfusion before or during tocolysis. The values of PI in umbilical artery, fetal descending aorta and middle cerebral artery did not reveal significant difference in all group. Nevertheless, 30% of pregnancies with preterm labor have pathological perfusion in the fetal descending aorta. We conclude that some patients with symptoms of preterm labor have pathological perfusion in uterine arteries and fetal descending aorta not induced by uterine contractions. We believe that the complex assessment of uterine perfusion should include the median PI value of both uterine arteries in order to have a reliable information. A short-term intravenous tocolysis with betamimetics does not seem to have a positive influence on the hemodynamic insufficiency.

The granulomatous skin lesions of human leprosy are known to be due to the cutaneous immune reaction to various mycobacterial antigens. In the present study, by immunohistochemical analysis using a previously characterized monoclonal antibody (MoAb) 3A8 we have demonstrated a selective expression of the 3A8 epitope of mycobacterial 30-kDa proteins, the major secreted proteins of mycobacteria, in various forms of leprosy lesions across the clinical spectrum. The localization of MoAb 3A8 staining is confined to the areas of cellular infiltrates of the lesions. In tuberculoid lesions the intense 3A8 staining was seen mostly in association with the membrane of the dermal cellular infiltrates whereas in highly bacilliferous lepromatous lesions the staining seems to be diffused with granular appearance but not in the form of bacteria. In patients with reversal reaction the staining was specifically extended to cells infiltrating the epidermis. MoAb 3A8 did not show any reactivity with inflammatory skin lesions of patients other than those with leprosy. Since the 3A8 epitope of 30-kDa proteins has been shown to be present in all cellular compartments of the mycobacteria and in the actively secreted BCG 85 antigen complex, MoAb 3A8 reactive protein(s) in leprosy lesions may be derived either from degraded somatic mycobacterial products or from antigens actively secreted by live bacilli. The latter could be true in the cases of untreated lepromatous lesions with high bacterial load since live M. leprae has also been considered to secrete corresponding 30-kDa proteins similar to other closely related mycobacteria. By double immunoenzyme staining we clearly demonstrate the expression of 3A8 epitope on CD68+ macrophages in the granulomas of tuberculoid leprosy, whereas in highly bacilliferous lepromatous lesions most of the double staining was seen in a diffuse pattern within the interstitial space of the cellular infiltrate as well as in the cytoplasm of CD68+ macrophages. In lesions from reversal reaction the 3A8 epitope is more strongly expressed on CDla+ dendritic Langerhans cells (LC) both in the epidermis and in the dermis as compared with other types of leprosy. This provides evidence for the involvement of LC in handling of mycobacterial antigenic epitopes in leprosy lesions. Further, immunoenzyme double staining revealed that the expression of this mycobacterial 3A8 epitope on antigen presenting cells such as CD68+ macrophages and CDla+ LC is present in juxtaposition with CD3+ T cells including the alpha beta and gamma delta receptor-bearing T cells in the granuloma.(ABSTRACT TRUNCATED AT 400 WORDS)

In 15 cases of singleton pregnancies with a sonographically supported diagnosis of intra-uterine growth retardation, the effects of maternal oxygen inhalation on uterine and feto-placental circulation have been investigated. For this purpose, the resistance and pulsitility indices (RI and PI) were measured in uterine artery placental site and uterine artery non-placental site, in the umbilical artery, in the descending thoracic aorta and in the median cerebral artery immediately before and after a half-hour oxygen inhalation in each of these IUR pregnancies. The median PI of uterine artery placental site decreased from 0.90 to 0.82 after oxygen inhalation, that of uterine artery non-placental site dropped significantly from 1.26 to 0.75 (p less than 0.05). The number of pathological findings in uterine artery non-placental site was significantly reduced (p less than 0.01). The median RI of the umbilical artery showed a minor decrease. Pathological perfusion in the umbilical artery was not noticeably affected either. The pathological median RI of the descending thoracic aorta was clearly reduced from 1.0 to 0.93. The median RI of the median cerebral artery showed a significant increase from 0.78 to 0.84 (p less than 0.05). The number of pathological findings was reduced by 50%. Compared with only two vessel indices (uterine artery non-placental site, umbilical artery) prior to oxygen inhalation, there was a significant correlation of all vessel indices with subsequent birth weight after oxygen inhalation.(ABSTRACT TRUNCATED AT 250 WORDS)

It was the aim of the present study to confirm and to pursue our previous nuclear medical investigations (Ruckhaberle et al. 1986) by using the Doppler ultrasound method to measure the uteroplacental, fetoplacental and fetal blood flow velocity waveforms in preterm labor. Apart from confirming impaired hemodynamics, we were especially interested to establish whether maternal oxygen inhalation could bring about any improvement of the impairment and result in a better pregnancy outcome.

The engineered disulfide bridge between residues 21 and 142 of phage T4 lysozyme spans the active-site cleft and can be used as a switch to control the activity of the enzyme (Matsumura, M. & Matthews, B.W., 1989, Science 243, 792-794). In the oxidized form the disulfide increases the melting temperature of the protein by 11 degrees C at pH 2. The crystal structure of this mutant lysozyme has been determined in both the reduced and oxidized forms. In the reduced form, the crystal structure of the mutant is shown to be extremely similar to that of wild type. In the oxidized form, however, the formation of the disulfide bridge causes the alpha-carbons of Cys 21 and Cys 142, on opposite sides of the active-site cleft, to move toward each other by 2.5 A. In association with this movement, the amino-terminal domain of the protein undergoes a rigid-body rotation of 5.1 degrees relative to the carboxy-terminal domain. This rotation occurs about an axis passing through the junction of the amino-terminal and carboxy-terminal domains and is also close to the axis that best fits the apparent thermal motion of the amino-terminal domain seen previously in crystals of wild-type lysozyme. Even though the engineered Cys 21-Cys 142 disulfide links together the amino-terminal and carboxy-terminal domains of T4 lysozyme, it does not reduce the apparent mobility of the one domain relative to the other. The pronounced "hinge-bending" mobility of the amino-terminal domain that is suggested by the crystallographic thermal parameters of wild-type lysozyme persists in the oxidized (and reduced) mutant structures. In the immediate vicinity of the introduced disulfide bridge the mutant structure is more mobile (or disordered) than wild type, so much so that the exact conformation of Cys 21 remains obscure. As with the previously described disulfide bridge between residues 9 and 164 of T4 lysozyme (Pjura, P.E., Matsumura, M., Wozniak, J.A., & Matthews, B.W., 1990, Biochemistry 29, 2592-2598), the engineered cross-link substantially enhances the stability of the protein without making the folded structure more rigid.

With the introduction of reproducible serological tests it was hoped that relapses in leprosy patients, after discontinuing treatment, could be detected before damaging reactions occurred and before the patients became infectious. The possible value of an ELISA using a semisynthetic analogue of phenolic glycolipid-I to detect antibodies to this antigen in order to predict a relapse in multibacillary patients was investigated. In contrast to that reported for paucibacillary patients, this test was useful to detect early relapses in multibacillary patients. In 3 out of 4 multibacillary patients who relapsed, the ELISA-values were increased. The decreased ELISA-values in the one relapsed patient could be attributed to the corticosteroid therapy. In the multibacillary patients who did not relapse after RFT, the ELISA-values were consistently low or decreased. In only one patient did the ELISA-values increase following his release from treatment and this patient was clinically suspected of developing a relapse.