Sealing of ruptured amniotic membranes with intra-amniotic platelet-cryoprecipitate plug.
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Hepatitis B virus (HBV) DNA contains consensus elements for transactivating proteins whose binding activity in other systems is regulated by inflammatory cytokines. Because HBV replicates within an environment of provoked inflammation, we speculated that the HBV core/pregenomic promoter may be regulated by cytokines produced in response to infection. To evaluate this hypothesis, the HBV core/pregenomic (C/P) promoter and associated cis-acting elements were placed upstream of a luciferase-encoding plasmid. This reporter construct was transfected into cytokine-sensitive hepatoma cells permissive for HBV replication, which were exposed to stimulated mononuclear cell-conditioned medium or human recombinant cytokines. Conditioned medium reduced luciferase expression by 80%. Tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), and interferon alfa (IFN-alpha) each reduced luciferase activity by 40%. Combinations of TNF-alpha and interferons mimicked the extent of conditioned medium inhibition. Non-specific effects from diminished cellular viability or growth were not responsible for decreased luciferase activity. Retention of HBV DNA 330 basepairs upstream of the C/P transcription start site was required to maintain the TNF-alpha effect. A 60% reduction in HBV replicative forms within intracellular core particles was demonstrated with TNF-alpha treatment of Hep G2 cells stably transfected with HBV DNA. The inhibitory action of these cytokines implicates a noncytolytic mechanism by which antigen-nonspecific immune responses in part regulate HBV replication in infected hepatocytes. This function may be beneficial in accelerating viral clearance, but in alternative circumstances could contribute to viral persistence by attenuating immunogen recognition.
OBJECTIVE: To compare the diagnostic and prognostic performance of maternal blood C-reactive protein, white blood cell count (WBC), and temperature with that of amniotic fluid (AF) WBC in preterm labor. METHODS: One hundred two women with preterm labor and intact membranes were studied. Maternal blood was collected to measure C-reactive protein concentration and WBC, and maternal temperature was also measured. Amniotic fluid obtained by amniocentesis was cultured and WBC determined. Receiver operating characteristic curve, logistic regression, and survival techniques were used for analysis. RESULTS: Patients with acute histologic chorioamnionitis had significantly higher median C-reactive protein concentration, WBC, temperature, and AF WBC than patients without this lesion (P < .05). Receiver operating characteristic curve and survival analysis demonstrated that an elevated C-reactive protein, WBC, or AF WBC was strongly associated with the likelihood of histologic chorioamnionitis, shorter interval to delivery, clinical chorioamnionitis, and neonatal morbidity (P < .05 for each). Of all the tests, AF WBC was the best independent predictor of a positive AF culture (odds ratio [OR] 16.8), interval to delivery (hazard ratio 5.7), clinical chorioamnionitis (OR 15.2), neonatal sepsis (OR 16.8), and significant neonatal complications (OR 7.4), after other confounding variables were adjusted (P < .05 for each). CONCLUSION: An elevated C-reactive protein, WBC, or AF WBC identified patients with intrauterine infection and adverse perinatal outcomes. Amniotic fluid WBC was a better independent predictor of these outcomes than C-reactive protein, WBC, or temperature.
The P3(00) event-related brain potential (ERP) was elicited with auditory and visual stimuli using an oddball task in which the subject indicated with a button-press response the occurrence of a target stimulus that occurred randomly on 20% of the trials and refrained from responding to a standard stimulus. A total of four trial blocks were collected for each stimulus modality condition. P3 amplitude from the target stimuli decreased reliably across trial blocks for both the auditory and visual stimulus condition, with no interaction obtained between stimulus modality and trial block. P3 latency was shorter for the auditory compared to visual stimulus conditions, but did not vary with trial block. No changes in amplitude or latency independent of the P3 effects were obtained for the other ERP components with trial block, although the usual modality differences were observed. The results suggest that P3 components elicited by auditory and visual stimuli both habituate for actively discriminated target stimuli. The theoretical implications are discussed in the context of previous findings.
OBJECTIVE: Periventricular leukomalacia, a common brain white matter lesion in preterm neonates, is a major risk factor for cerebral palsy. Recently, cytokines (i.e., tumor necrosis factor and interleukin-1(beta)) have been implicated as mediators for the development of periventricular leukomalacia. The purpose of this study was to examine the relationship between umbilical cord plasma levels of tumor necrosis factor-alpha, interleukin-1(beta), interleukin-6, and interleukin-1 receptor antagonist and the occurrence of periventricular leukomalacia in preterm neonates. STUDY DESIGN: Umbilical cord blood was collected from 172 consecutive preterm births (25 to 36 weeks). Periventricular leukomalacia-associated lesions were diagnosed by brain ultrasonography within the first 3 days of life. Tumor necrosis factor-alpha, interleukin-1(beta) interleukin-6, and interleukin-1 receptor antagonist were measured by sensitive and specific enzyme-linked immunoassay methods. Umbilical cord arterial pH was measured at birth. Statistical analysis was performed with multiple logistic regression and receiver operating characteristic curve analysis. RESULTS: Periventricular leukomalacia-associated lesions were present in 14.5% (25/172) of infants. Plasma concentrations of interleukin-6 but not of tumor necrosis factor-alpha, interleukin-1(beta), and interleukin-1 receptor antagonist were significantly higher in neonates with periventricular leukomalacia-associated lesions than in those without these lesions (median 718, range < 226 to 32,000 pg/ml vs median < 226, range < 226 to 43,670 pg/ml; p < 0.0001). An interleukin-6 value > or = 400 pg/ml had a sensitivity of 72% (18/25) and a specificity of 74% (108/147) in the identification of periventricular leukomalacia-associated lesions. Multivariate analysis showed that umbilical cord interleukin-6 was an independent risk factor for periventricular leukomalacia (odds ratio 6.2, p < 0.002) after correction for known confounding variables (i.e., gestational age at birth, umbilical artery pH, chorioamnionitis). CONCLUSIONS: Interleukin-6 concentrations in umbilical cord plasma are elevated in neonates with periventricular leukomalacia-associated lesions. Our data support the hypothesis that periventricular leukomalacia may be the result of cytokine-mediated brain injury.
OBJECTIVES: The response to infection by human first-trimester trophoblasts is a poorly understood event. This study was undertaken to determine whether first-trimester trophoblasts are capable of responding to an infection stimulus and mediating an immune response. STUDY DESIGN: HTR-8/SVneo cells were exposed to lipopolysaccharide (1 microgram/ml) or media alone for either 0, 2, 4, 6, 8, or 24 hours. Northern analysis was conducted by use of a panel of antisense cytokine probes. Enzyme-linked immunosorbent assays specific for either interleukin-1 alpha, interleukin-6, interleukin-8, or transforming growth factor-beta 1 were conducted on corresponding cell culture supernatants, and the kinetics of expression were determined. RESULTS: Interleukin-1 alpha, interleukin-6, interleukin-8, and transforming growth factor-beta 1 transcription occurred maximally between 2 and 8 hours of culture in media containing lipopolysaccharide, with a subsequent diminution of response. Enzyme-linked immunosorbent assay analysis corroborated lipopolysaccharide induction seen at the level of transcription, with significant posttranslational expression of these cytokines being detected between 2 and 24 hours in culture (p < 0.01). CONCLUSIONS: Expression of the proinflammatory cytokines interleukin-1 alpha, interleukin-6, interleukin-8 and transforming growth factor-beta 1 strongly support the contention that human first-trimester trophoblasts are capable of responding to an infection stimulus and eliciting an immune response through cytokine-based immune signaling.
BACKGROUND: PEG disruptions during conversions to skin-level gastrostomy devices have been described, but specific risk factors have not been reported. In this study, possible risk factors for tract disruption in a pediatric population were identified, and management of complications described. METHODS: The medical records of patients who underwent gastrostomy conversions during 1994 were reviewed. Statistical analysis was performed using two-tailed student's t test, and risk ratios with 95% confidence limits were calculated. RESULTS: Gastrostomy tract disruption occurred in 6 to 30 (20%) of tube conversions. Complicated and uncomplicated cases did not differ with regard to age, sex, primary or associated diagnoses, pregastrostomy or postgastrostomy nutritional status, tract maturity, or percutaneous gastrostomy tube type. The use of an 18F obturator-type skin-level gastrostomy tube increased the risk for gastric separation 4.8-fold. Tract disruptions were managed by fluoroscopic gastrostomy tube replacement, repeat PEG, or exploratory laparotomy with open gastrostomy. CONCLUSIONS: The use of obturator-type skin-level gastrostomy tubes was associated with an increased risk of tract disruption. Fluoroscopic verification of intragastric placement is warranted after initial conversions to skin-level gastrostomy tubes.
OBJECTIVE: To compare the diagnostic performance of maternal blood C-reactive protein, white blood cell count (WBC), and amniotic fluid (AF) WBC in the identification of positive AF culture, histologic and clinical chorioamnionitis, and neonatal morbidity in women with preterm premature rupture of membranes (PROM). METHODS: Maternal blood was collected for the determination of C-reactive protein and WBC at the time of amniocentesis from 90 women with preterm PROM. Amniotic fluid was cultured for aerobic and anaerobic bacteria as well as mycoplasmas. Amniotic fluid WBC was determined for research purposes. Receiver operating characteristic curve and logistic regression were used for statistical analysis. RESULTS: The prevalence of positive AF culture was 28% (25 of 90). Women with positive AF culture and clinical chorioamnionitis had significantly higher median C-reactive protein, WBC, and AF WBC than did women without these conditions (P < .05), whereas women with histologic chorioamnionitis and significant neonatal morbidity had higher median C-reactive protein and AF WBC, but not WBC, than those without the conditions (P < .05). An AF WBC of at least 20 cells per mm3 had a greater sensitivity than C-reactive protein (cutoff, 0.7 mg/dL) and WBC (cutoff, 13,000 cells per mm3) in the detection of positive AF culture and histologic chorioamnionitis. Logistic regression analysis indicated that among AF WBC, C-reactive protein, and WBC, AF WBC was the best predictor of positive AF culture (odds ratio [OR] 24.2, 95% confidence interval [CI] 6.0, 97.5, P < .001), histologic (OR 74.0, 95% CI 7.4, 736.3, P < .001) and clinical chorioamnionitis (OR 8.9, 95% CI 0.9, 85.6, P = .057), and neonatal morbidity (OR 4.3, 95% CI 1.1, 16.6, P < .05). CONCLUSION: Amniotic fluid WBC performs better than C-reactive protein and maternal blood WBC in the diagnosis of positive AF culture, histologic and clinical chorioamnionitis, and neonatal morbidity in women with preterm PROM.
The traditional paradigm that prostaglandins (PGS) are of central importance in the initiation of labor has been challenged. A group of investigators has recently reported that the amniotic fluid concentrations of PGE(2) and PGF(2 alpha) increase only late in the course of labor implying that "the accumulation of prostaglandins in amniotic fluid is an after-effect of labor and not indicative of a role of these compounds in the initiation of human parturition." The present study was conducted to determine whether amniotic fluid prostaglandin concentrations increase prior to the onset of human labor, the central question in this controversy. Three amniocenteses were performed in 17 women with intrahepatic cholestasis of pregnancy -- the first two prior to the onset of labor and the third during early spontaneous labor. PGE(2) and PGF(2 alpha) were measured with sensitive and specific radioimmunoassays. Amniotic fluid concentrations of PGE(2) and PGF(2 alpha) increased prior to the onset of spontaneous labor. An additional increase in the concentrations of PGE(2) and PGF(2 alpha) was found in samples obtained in early labor. We conclude that an increase in prostaglandin bioavailability precedes the onset of spontaneous human parturition.
The study was designed to report our experience with in utero percutaneous umbilical cord ligation for selective feticide in pre-viable, abnormal, monochorionic multiple gestations. The selection criteria prior to offering percutaneous umbilical cord ligation included normal anatomy in the other fetus(es), normal karyotype and polyhydramnios, or uterine contractions. The procedure was performed under combined endoscopic and sonographic guidance. Fifteen patients were referred for evaluation and possible treatment. Two patients were excluded because of misdiagnosis. The mean gestational age at the time of the procedure was 21 weeks (range 16-25 weeks). There were ten acardiac twins, one discordant twin with acrania, and two cases of twin-twin transfusion syndrome with a hydropic twin. Percutaneous umbilical cord ligation was accomplished in 11/13 (84%) cases. An average of 8.5 weeks (range 0-18.5 weeks) was gained after percutaneous umbilical cord ligation, and five of 11 patients delivered at or after 30 weeks. Seven of 11 patients undergoing ligation procedures had living children. Premature rupture of membranes occurred within 3 weeks in 4/13 (30%) of cases. In utero fetal demise remote from the procedure occurred in three ligated patients undergoing ligation, in two of which intraoperative bleeding occurred. One fetus with cystic fibrosis died in the newborn period. Percutaneous umbilical cord ligation is an evolving alternative technique for the management of abnormal monochorionic multiple gestations. The advantages of percutaneous umbilical cord ligation over other approaches include complete extravascular interruption of the blood flow in all vessels of the cord. This may prevent postmortem feto-fetal hemorrhage or possible embolic phenomena that may result in death of the normal twin, as seen with intravascular techniques. Percutaneous umbilical cord ligation prior to spontaneous death of a co-twin may also avert the neurological and other complications that may occur in the surviving twin. Technological advances should decrease the observed complications and improve the success rate of the procedure.
Clinically significant chorioangiomas, i.e. those greater than 4 cm in diameter, are associated with an overall fetal loss rate of approximately 40%. Maternal complications may also be present. Most patients diagnosed with chorioangiomas have been managed expectantly. A patient with a large chorioangioma was referred to our institution, with a fetus that was hydropic and showed sonographic signs of over heart failure. Fetal anemia, marked erythropoiesis and hypoalbuminemia were documented on cordocentesis. The blood supply to the mass consisted of an artery and a vein of 9 mm in diameter. The vascular supply to the tumor was ablated via operative fetoscopy by suture ligation of the arterial supply, after subchorionic dissection of the vessel. The remaining blood supply was electrocoagulated with bipolar cautery. Although the procedure was technically successful, the fetus died on the 3rd postoperative day. This case illustrates ablation of the blood supply of placental chorioangiomas as a potential management alternative for those patients with large chorioangiomas. Early identification and treatment may result in a successful outcome.
BACKGROUND: There are no specific recommendations for management of dyslipidaemia in patients with chronic renal failure (CRF) on haemodialysis, in which atherosclerosis is a common cause of morbidity and mortality. The aim of the present study was to analyse different approaches based on low-density lipoprotein (LDL) cholesterol (measured and calculated with a formula), non-high-density lipoprotein (HDL) cholesterol, and HDL cholesterol levels in the clinical management of dyslipidaemia in haemodialysis patients. METHODS: Calculated LDL cholesterol by the Friedewald formula was compared with measured LDL cholesterol after separation by ultracentrifugation in 101 male patients with CRF on haemodialysis and in 101 healthy control subjects. RESULTS: Calculated LDL cholesterol coincided with measured LDL cholesterol, with less than 10% error, in 54 patients (53.4%) and in 75 controls (74.2%). Calculated LDL cholesterol was overestimated, with an error of 10% or more with respect to measured LDL cholesterol, in 37.6% of patients and in 23.7% of controls, and underestimated in 8.9% and 1. 9% respectively. Despite a good correlation between calculated and measured LDL cholesterol, the intraclass correlation coefficients demonstrate a poor concordance between calculated and measured LDL cholesterol, both in patients and controls. Only 17 patients were at non-HDL cholesterol level risk defined as higher than 4.28 mmol/l. HDL cholesterol levels lower than 0.9 mmol/l were found in 70% of patients and in 23% of controls. CONCLUSIONS: LDL or non-HDL cholesterol levels may not be appropriate for management of lipoprotein abnormalities in CRF patients. Further studies must clarify whether HDL cholesterol may be the best lipoprotein parameter for evaluating cardiovascular risk in these patients.
A PCR-based sequence-tagged site (STS) content mapping strategy has been used to generate a physical map with 90% coverage of the 120-Mb euchromatic portion of the Drosophila genome. To facilitate map completion, the bulk of the STS markers was chosen in a nonrandom fashion. To ensure that all contigs were localized in relation to each other and the genome, these contig-building procedures were performed in conjunction with a large-scale in situ hybridization analysis of randomly selected clones from a Drosophila genomic library that had been generated in a P1 cloning vector. To date, the map consists of 649 contigs with an STS localized on average every 50 kb. This is the first whole genome that has been mapped based on a library constructed with large inserts in a vector that is maintained in Escherichia coli as a single-copy plasmid.
PROBLEM: GRO alpha/MGSA is a new member of the chemokine superfamily CXC(alpha) and is produced by a variety of cells including macrophages, fibroblasts, epithelial, and endothelial cells, and keratinocytes. This chemokine has chemoattractant activity and may participate in neutrophil recruitment and activation during the course of intrauterine infection. This study was conducted to investigate the effect of labor and microbial invasion of the amniotic cavity (MIAC) on amniotic fluid, fetal, and maternal plasma GRO alpha concentrations. METHOD: A cross-sectional study was designed using parameters that included gestational age, results of amniotic fluid (AF) cultures, and labor status at the time of amniocentesis. Fluid was retrieved by transabdominal amniocentesis. MIAC was defined as a positive amniotic fluid culture for bacteria. Umbilical cord blood was retrieved at the time of delivery. Amniotic fluid, maternal and fetal plasma GRO alpha concentrations were measured with a sensitive and specific ELISA (Quantikine, R&D Systems, Minneapolis, MN). RESULTS: 1) GRO alpha was detectable in amniotic fluid, umbilical cord, and maternal plasma samples; 2) GRO alpha concentrations in amniotic fluid increased with advancing gestational age; 3) Both term and preterm gestations with MIAC were associated with higher amniotic fluid GRO alpha concentrations than those with sterile amniotic fluid, independent of the labor status (term, MIAC, labor: median 2.7 ng/ml, range 1.4-12.7 vs. term, no MIAC, labor: median 2.1 ng/ml, range 0.7-3.4, vs term, no MIAC, no labor: median 1.9 ng/ml, range 1.8-4.2; P < 0.005; preterm: MIAC median 5 ng/ml, range 0.6-47.9 vs. no MIAC: median 2.3 ng/ml, range 0.5-10; P < 0.008); 4) A strong correlation was found between umbilical cord plasma GRO alpha concentrations and neonatal neutrophil count, and between GRO alpha concentrations and white blood cell count in the amniotic fluid (r = 0.67, P < 0.0005 and r = 0.38, P < 0.001, respectively). CONCLUSION: GRO alpha is a physiologic constituent of amniotic fluid and cord blood. Amniotic fluid GRO alpha concentrations increase with gestational age. Intrauterine infection both preterm and at term is associated with an increase in GRO alpha concentrations of amniotic fluid, suggesting that GRO alpha may play an important role in recruitment of neutrophils into the amniotic cavity.
PROBLEM: Comparatively little is known about the capacity of first trimester trophoblasts to respond to an infection and coordinate an immune response. This study characterizes the LPS induction of G-CSF and RANTES in a first trimester trophoblast cell line. METHODS: HTR-8/SV neo cells were exposed to LPS (1 micrograms/ml) for 0, 2, 4, 6, 8, and 24 hours in DMEM-Ham's F-12 media supplemented with 10% fetal bovine serum. Cytokine levels in culture supernatants were measured by ELISA. Northern analysis of total RNA was conducted using antisense cytokine probes. RESULTS: Levels of immunoreactive G-CSF and RANTES from LPS induced cultures at 24 hours were 10-fold and 8.5-fold greater than cytokine levels from non-induced cells at 24 hours, respectively (P < 0.01). Under LPS induction, maximal rates of G-CSF and RANTES transcription occurred at 24 hours and 8 hours, respectively. CONCLUSION: The LPS induction of proinflammatory cytokines in a first trimester trophoblast cell line supports the contention that first trimester trophoblasts participate in cytokine based immune signaling in response to infection.
Isolated magnesium malabsorption (congenital hypomagnesemia) has been reported in approximately 30 patients worldwide. Patients typically present by 6 months of age with convulsions and diarrhea. We report an unusual case of isolated magnesium malabsorption in an older boy with no diarrhea.
We report our experience of two years (February 1993-February 1995) in the endoscopic treatment with self-expanding prosthesis (Ultraflex) of 21 patients with inoperable malignant oesophageal stenosis. 23 prosthesis were implanted enabling swallowing in less than 24 h. There were no major complications. In two occasions the prosthesis was obstructed by tumoral growth needing replacement. The prosthesis was occluded by solid food in four cases, which were resolved endoscopically.
BACKGROUND: Non-immune fetal hydrops is a heterogeneous disorder with a mortality rate of 50-98%. Resolution of non-immune fetal hydrops is rare but has been reported to occur spontaneously or after targeted therapeutic measures. CASE: A euthyroid gravida with Graves disease presented with a history of three prior perinatal deaths between 26 and 28 weeks' gestation, all associated with fetal hydrops. In the current pregnancy, the fetus developed hydrops at 24 weeks' gestation. Fetal hyperthyroidism, with high-output cardiac failure, was diagnosed with fetal blood sampling. After maternal therapy with propylthiouracil, resolution of the non-immune hydrops were documented and a healthy neonate subsequently delivered to term. The neonate developed transient hyperthyroidism after delivery, which required treatment for 10 weeks. CONCLUSION: Non-immune hydrops occurring as a result of fetal hyperthyroidism with high output cardiac failure is treatable with propylthiouracil.