Detection of circulating placental protein 5 in patients with trophoblastic tumors.
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A two site solid phase radioimmunoassay for detection of common food antigens is described. Bovine serum albumin, beta-lactoglobulin and ovalbumin can be detected in normal human serum at levels ranging from 0.1 to > 1000 ng/ml; sensitivity is not impaired by the presence of low levels of antibodies. Thirty min to 3 h after oral intake of milk, beta-lactoglobulin could be detected in the sera of 3 normal individuals, at a concentration of 0.1-3 ng/ml. This assay should prove useful in assessing the importance of macromolecular absorption in food allergy and in other gastrointestinal diseases.
Serum obtained from healthy volunteers 6-7 h after consumption of 60-95 g of ethanol contained cytotoxic activity against mouse A9 cells and all of six human cell lines tested. Affinity chromatography of such sera demonstrated that at least some of the cytotoxic molecules consisted of altered albumin. Complexes formed by the reaction of 14C-acetaldehyde with 125I-labelled human serum albumin in vitro were also cytotoxic. After treatment with a reducing agent, sodium borohydride, the cytotoxicity of both post-alcohol serum and the acetaldehyde-albumin complexes fell sharply, suggesting that the cytotoxic activity resided in the unstable Schiff bases formed during the first stage of reaction between the acetaldehyde and proteins. A detailed analysis of the reaction between the double-labelled acetaldehyde-albumin complexes and K562 cells revealed that the cytotoxic activity resulted from the release of acetaldehyde from such complexes and the preferential binding of the free acetaldehyde to the target cells.
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The extended interaction of aldoses with proteins or lipids results in nonenzymatic glycation and oxidation, ultimately forming AGEs, the presence of which in the plasma and vessel wall is associated with diabetic vascular complications. We show here that AGE albumin in the intravascular space interacts with the vessel wall via binding to an integral membrane protein, receptor for AGE (RAGE), a member of the immunoglobulin superfamily, resulting in clearance from the plasma and induction of interleukin 6 mRNA. Intravenously infused 125I-AGE albumin showed a rapid phase of plasma clearance with deposition in several organs. Rapid removal of 125I-AGE albumin from the plasma was prevented by administration of a soluble, truncated form of RAGE, which blocked binding of 125I-labeled AGE albumin to cultured endothelial cells and mononuclear phagocytes, as well as by pretreatment with anti-RAGE IgG. Ultrastructural studies with AGE albumin-colloidal gold conjugates perfused in situ showed that in murine coronary vasculature this probe was taken up by endothelial plasmalemmal vesicles followed by transport either to the abluminal surface or by accumulation in intracellular vesicular structures reminiscent of endosomes and lysosomes. Consequences of AGE-RAGE interaction included induction of interleukin 6 mRNA expression in mice. These data indicate that RAGE mediates the interaction of AGEs with the vessel wall, both for removal of these glycated proteins from the plasma and for changes in gene expression.
Plasma volume and plasma protein were measured on days 5, 12, 19, and 21 of gestation in rats fed 25% casein either ad libitum or 55% food restricted, or fed 6% casein ad libitum (protein restricted). Both forms of undernutrition prevented the large increase in plasma volume observed in well-fed rats by day 19. The pattern of plasma volume change, with maximum mean values on day 19 (compared to the other measurement days), was not altered by undernutrition. Differences in plasma protein concentration or total circulating protein did not explain plasma volume changes.
OBJECTIVE: To investigate protein complement 3a (C3a) and protein complement 3 (C3) plasma levels in trauma patients directly after the injury, in relation to the patients' outcome, the development of sepsis, or the injury severity, as determined by either the Polytrauma Score (PTS), the Injury Severity Score (ISS), or the Trauma and Injury Severity Score (TRISS). DESIGN: Prospective study. SETTING: Surgical intensive care unit in a university hospital. PATIENTS: Thirty-four patients with multiple trauma. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: C3a and C3 concentrations, as well as the C3a/C3 ratio, were determined at the time of the accident (T0), at the emergency admission (T1), 8 hrs after the accident (T2), and every 8 hrs until day 3, every 12 hrs until day 6, and once daily on days 7 and 8. The C3a plasma concentrations and the C3a/C3 ratios of nonsurvivors were significantly greater at T0 or T1 as compared with those survivors (p = .008 or .033). Patients who developed sepsis had higher C3a plasma levels at the scene of accident than patients without complications. However, these differences did not reach statistical significance (p = .051), although a clear trend was apparent. Patients were grouped according to the severity of injury, as determined by either the PTS, ISS, or TRISS. We found significant differences in the both the mean C3a values and the C3a/C3 ratio among the different groups, during the first 8 hrs after the injury. In addition, a significant correlation was observed between the C3a concentration or the C3a/C3 ratio at T0 to T2 and either the ISS (r2 = .49), PTS (r2 = .22) or the TRISS (r2 = .45), which was similar to correlations between injury severity scores themselves (r2 = .36 to .58). CONCLUSIONS: Complement activation occurs immediately after the injury. The degree of activation is a hallmark for the outcome of a patient. Determination of C3a concentrations, at the scene of the accident, may prove helpful to assess the severity of the injury and to determine the prognosis. The amount of C3a and the C3a/C3 ratio may be useful as additional parameters to the existing trauma scoring systems, such as, PTS, ISS, and TRISS.
The clinical scoring systems of atopic dermatitis were analysed and compared. Some biological parameters that can correlate with the clinical score were also reviewed. After the definition of the disease based on validated clinical criteria, the second necessity was the availability of reliable severity scores to allow clinicians to verify the course of the disease and the efficacy of treatments. After many proposals, the SCORAD (SCORing Atopic Dermatitis), that required more than three years of work, was the first one that was validated. SCORAD is freely available from an internet site and can be easily calculated using dedicated software. EASI (Eczema Area and Severity Index) score has also been validated but it has been modified twice. Simpler systems include SASSAD (Six Area, Six Sign Atopic Dermatitis) and TIS score (Three-Item Severity score). In parallel, biological parameters were investigated. Eosinophil cationic protein, circulating basophils, major basic protein, soluble E-selectin, antistaphylococcal enterotoxin B, immunoglobulin E titres and macrophage-derived chemokine, can correlate significantly with the clinical score. The clinicians will not benefit directly from laboratory techniques and will employ clinical scores.
Serum human chorionic gonadotrophin (hCG), pregnancy-specific beta 1-glycoprotein (SP 1) and placental proteins 5 (PP5) levels have been measured by radioimmunoassay in 20 patients (116 samples) with hydatidiform mole, one patient (nine samples) with invasive mole and 10 patients (103 samples) with choriocarcinoma. Measurement of both serum SP1 and hCG are useful in the monitoring of these diseases. The presence of PP5 in hydatidiform mole and its absence in choriocarcinoma support the hypothesis that PP5 is closely associated with the invasive activity of malignant trophoblast.
Maternal serum levels of human chorionic gonadotrophin (hCG), Schwangerschaftsprotein 1 (SP1) and pregnancy-associated plasma protein A (PAPP-A) were measured in an unselected group of 624 women presenting with amenorrhoea and vaginal bleeding with or without abdominal pain to an emergency gynaecological ultrasound clinic. Abdominal sector scanning was used to assess uterine contents. Pregnancy was confirmed by ultrasound in 406 pregnancies. Histological confirmation was obtained in each case of pregnancy failure. A live fetus was demonstrated in 259 women of whom six subsequently miscarried; one of these had markedly depressed serum hCG and PAPP-A, but normal SP1 levels, and two had oligohydramnios. Of the 147 women without ultrasound evidence of fetal heart action 67 had a correct ultrasound diagnosis of anembryonic pregnancy. The predictive value of a depressed serum hCG level was 70% in this group, and 31% in samples taken at less than or equal to 7 weeks. The predictive value of a normal hCG level was 96%. In 34 women missed miscarriage was diagnosed readily by ultrasound; all but five had depressed hCG and PAPP-A levels. A clinical diagnosis of a complete or incomplete miscarriage was made in 45 women and easily confirmed by ultrasound. All of them had depressed hCG, SP1 and PAPP-A levels. These results indicate that the diagnostic value of ultrasound in threatened miscarriage is often better than that of biochemical tests.
The kinetics and nature of the nondialyzable cytotoxic activity which appeared in the serum after the consumption of 1.2 g ethyl alcohol per kilogram body weight over 45 min was studied in six healthy volunteers and eight patients with histologically proven alcohol-related cirrhosis of the liver. Whereas the cytotoxic activity in the dialyzed serum showed a single peak with a maximum value 8 hr after the start of ethanol consumption in the healthy volunteers, it showed two peaks with maximum values at 2 and 8 hr in the patients with cirrhosis. Studies of the fractions obtained by Sephacryl-S-300 gel filtration of the 2-hr postalcohol serum samples revealed substantial cytotoxic activity in the fractions containing both the albumin peak and the IgG peak in the patients with cirrhosis and only in the fractions containing the albumin peak in the healthy volunteers. Experiments with pure IgG preparations obtained from prealcohol and 2-hr postalcohol sera by chromatography on Q-Sepharose Fast Flow anion-exchange resin showed considerable cytotoxic activity in the preparations from the patients with cirrhosis and little or no cytotoxic activity in those from the healthy volunteers. Thus, the early peak of the biphasic serum cytotoxicity curve seen after ethanol consumption by patients with cirrhosis appeared to be caused by the development of a substantial cytotoxic activity in the IgG molecules during the first 2 hr.(ABSTRACT TRUNCATED AT 250 WORDS)
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The systemic inflammatory response of the body to invading microorganisms, termed sepsis, leads to profound activation of the complement system. Pathophysiological concepts suggest that complement activation occurs very early in this syndrome. Thus, we discuss whether the determination of concentrations of the complement components C3a, C5a, and C3 in plasma as well as of the C3a/C3 ratio might be helpful to diagnose sepsis early. For this purpose, 33 patients from an intensive care unit were monitored for 10 days. In comparison with healthy donors, C3a levels and the C3a/C3 ratio of intensive-care-unit patients were significantly elevated (P < 0.0001) on admission. In contrast, C3 levels were significantly reduced (P < 0.0001) but increased during the study. C5a levels in the plasma of healthy donors and patients were identical. Twenty-two of 33 patients fulfilled microbiological and clinical criteria of sepsis. Eleven patients had signs of systemic inflammatory response syndrome but no microbiological evidence of sepsis. The groups could be differentiated from each other by their C3a levels or their C3a/C3 ratios during the first 24 h after the clinical onset of sepsis (P < 0.05). Septic patients in shock had higher C3a levels than normotensive septic patients, although the differences were not significant. Nonsurvivors had significantly higher C3a levels on admission than survivors (P = 0.0185). No differences were found between septic patients who developed adult respiratory distress syndrome and those who did not. Thus, determination of C3a concentrations in plasma may prove useful (i) to diagnose sepsis early, (ii) to differentiate between patients with sepsis and those with systemic inflammatory response syndrome, and (iii) to assess prognosis.
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We investigated expression of the adipose-specific serine protease adipsin in genetically obese Zucker rats and whether adrenalectomy modifies expression. Adipsin mRNA levels were determined by slot blot and Northern blot analysis of total RNA samples extracted from epididymal adipose tissue and isolated retroperitoneal adipocytes of obese (fa/fa) and homozygous lean (Fa/Fa) Zucker rats. Both 30-day-old and 4-mo-old animals were analyzed in experiment 1. In experiment 2, 10-wk-old obese and lean rats were either bilaterally adrenalectomized or sham operated, and adipsin mRNA levels were determined on tissue and cell samples 2 wk postsurgery. In both experiments, serum adipsin protein was determined by Western blot analysis and plasma insulin by radioimmunoassay. The data show that both adipsin mRNA and adipsin protein are reduced in obese compared with lean rats and that adrenalectomy restores these values toward normal in obese rats. The data thus suggest that adrenal steroids are involved in modulating adipsin expression in obese Zucker rats and that insulin may be an intermediary factor in such modulation.
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Fetal and neonatal ovine plasma protein and lactate dehydrogenase concentrations have been measured between 124 and 146 days gestation in th chronically catheterized fetal sheep and in the newborn lamb. Fetal plasma protein concentration ranged from 24.7 to 77.0 mg . ml-1 and increased at a rate of 1.65 mg . ml-1 . day-1. In contrast, in the first 10 days of neonatal life plasma protein concentration fell steadilyu at a rate of 2.64 mg . ml-1 . day-1. Fetal plasma lactate dehydrogenase concentration increased at a rate of 0.014 IU . ml-1 . day-1 during the period of study. The increase correlated significantly with the rise of fetal plasma protein concentration. In contrast, in the newborn lamb no significant change in plasma lactate dehydrogenase concentration was observed during ther first 10 days after birth.
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