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Serum growth factors cause rapid stimulation of protein synthesis and dephosphorylation of eIF-2 in serum deprived Ehrlich cells.

In Ehrlich ascites tumor cells maintained in serum-free medium for 16 h the rate of protein synthesis was about 50% of the rate in control (well-fed) cells. The addition of 10% calf serum led to a 1.5- to 2-fold stimulation of protein synthesis within 10 min. Stimulation was effected through a non-transcriptional mechanism which operated at the level of polypeptide chain initiation. The effect was due to non-dialyzable serum growth factors which were sensitive to treatment with dithiothreitol and iodoacetamide. Replacing the 16-h-conditioned serum-free medium with fresh serum-free medium stimulated protein synthesis about 30% in serum-deprived cells, and the effect of these low molecular weight nutrients was additive with the effect of serum factors. Phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF-2 alpha) inhibits protein synthesis by competitively inhibiting the guanine nucleotide exchange factor (GEF), and modulation of the extent of phosphorylation of eIF-2 alpha has been suggested as a probable regulatory mechanism in serum-deprived mammalian cells. We measured the ratio of phosphorylated to total eIF-2 alpha in serum-deprived cells. The ratio was elevated in serum-deprived cells compared to control (serum-fed) cells. eIF-2 was rapidly dephosphorylated in response to serum refeeding and returned to near control levels after 10 min. The rapidity of this response and the close temporal correlation between eIF-2 dephosphorylation and increased rate of protein synthesis provide evidence that eIF-2 plays an important role in the regulation of protein synthesis by serum growth factors.

Animals↗

Prospective study on anti-hepatitis C virus-positive patients with persistently normal serum alanine transaminase with or without detectable serum hepatitis C virus RNA.

A significant proportion of patients with detectable antibodies to hepatitis C virus have normal serum alanine transaminase levels. Our aim was to study the outcome of this group. Between 1992 and 1999, 135 consecutive anti-HCV-positive patients with persistently normal ALT were followed for 3.6 +/- 2.3 years (0.5 to 8.5 years), 108 had a liver biopsy at inclusion, and 24 had a second liver biopsy 3.5 +/- 1.0 years later. Serum HCV RNA was detectable with PCR in 94 patients (69%) and not detectable in 41 patients (31%). Patients with and without detectable serum HCV RNA had similar epidemiological characteristics. Serum ALT levels and anti-HCV ratio were lower (P =.001), and histological lesions had lower grade and stage in patients without detectable serum HCV RNA (P =.001). Liver HCV RNA was not detectable with PCR in the 12-serum HCV RNA-negative patients tested. During follow-up, all patients without detectable serum HCV RNA remained HCV RNA-negative and kept normal serum ALT; all patients with detectable serum HCV RNA remained HCV RNA-positive, 20 (21%) had a slight fluctuation of serum ALT above the upper limit of normal. No significant changes were observed in the liver lesions of the 24 patients who underwent a second liver biopsy. In anti-HCV-positive patients with persistently normal serum ALT, histological lesions are significantly lower in HCV RNA-negative than in HCV RNA-positive patients. During follow-up, the HCV RNA status of patients remained unchanged; 21% of the patients with detectable serum HCV RNA had slight increase in serum ALT levels, but histological lesions remained stable.

Adult↗

Relation of serum ascorbic acid to serum lipids and lipoproteins in US adults.

OBJECTIVE: To examine the relation of serum ascorbic acid level to serum lipid and lipoprotein levels among a random sample of the US adult population. METHODS: Using linear regression, the relation of serum ascorbic acid level to serum lipid and lipoprotein levels was examined among 5,412 women and 5,116 men enrolled in the Second National Health and Nutrition Examination Survey (NHANES II), 1976-1980. Age, race, body mass index, level of physical activity, level of education, alcohol intake, and dietary energy, cholesterol, and fat intakes, and other potential confounders were included in the multivariate models. RESULTS: Serum ascorbic acid level was independently associated with high-density lipoprotein cholesterol (HDL-C) among women; each 1 mg/dl increase in serum ascorbic acid level (range 0.1 to 2.7 mg/dl) was associated with a 2 mg/dl increase in HDL-C level (p = 0.001). Because other investigators have demonstrated an inverse relation between ascorbic acid intake or blood levels and total serum cholesterol in individuals with elevated total serum cholesterol levels, we analyzed four subgroups of NHANES II participants with total serum cholesterol levels > 200 mg/dl. Among women with total serum cholesterol levels > or = 200 mg/dl, each 1 mg/dl increase in serum ascorbic acid level was independently associated with an increase of 2 to 3 mg/dl in HDL-C level (p < or = 0.05). Serum ascorbic acid level was not significantly associated with other serum lipids or lipoproteins. CONCLUSIONS: If the observed associations are linked causally, they would suggest that ascorbic acid is a factor in cholesterol homeostasis among women and may be particularly important for women at increased risk for coronary heart disease.

Adolescent↗

Increased endothelial monolayer permeability is induced by serum from women with preeclampsia but not by serum from women with normal pregnancy or that are not pregnant.

OBJECTIVE: To determine if endothelial monolayer permeability could be altered by serum from preeclampsia (PE). METHODS: Confluent normal endothelial cells (ECs) were incubated with 20% serum from nonpregnant females, normal and PE pregnancies or combined with antioxidant superoxide dismutase (SOD) for 8 hr. Confluent PE ECs were incubated with 20% serum from normal pregnancies. EC barrier function of monolayer permeability was accessed by measuring EC electrical resistance (ER) and the leakage of horseradish peroxidase (HRP) passing through EC filters. Plasma concentrations of IL-8 and lipid peroxides by MDA were also measured. We determined 1) if serum from PE could affect EC permeable function; 2) if antioxidant and serum from normal pregnancies could preserve PE EC barrier function; 3) if lipid peroxides and cytokine IL-8 were increased in PE blood samples. Data are presented as mean+/-SE. ANOVA was used for statistical analysis. A p level less than 0.05 was considered statistically different. RESULTS: 1) ER was significantly decreased and HRP passage was significantly increased in ECs incubated with serum from PE compared to serum from non-pregnant and normal pregnant females (ER: 36.30+/-2.60 vs. 51.30+/-4.00 and 53.90+/-5.80 Omega x cm2, p<0.01; HRP: 0.100+/-0.020 vs. 0.014+/-0.002 and 0.022+/-0.007 DeltaOD470 nm, p<0.01, respectively). 2) ER was improved in PE ECs incubated with serum from normal pregnancies compared to controls, 52.28+/-3.13 vs. 34.50+/-3.80 Omega x cm2, p<0.01. 3) SOD attenuated decreased EC ER induced by PE serum, 55.58+/-3.61 Omega x cm2 (SOD+PE serum) vs. 42.34+/-3.24 (control) and 35.46+/-2.44 (PE serum), p<0.01, respectively. 4) Both MDA and IL-8 concentrations were higher in plasma or serum samples from PE than those in samples from nonpregnancies and normal pregnancies, MDA: 28.65+/-1.45 vs. 22.40+/-1.47 and 25.53+/-0.89 micromol/mL, p<0.01; IL-8: 5.35+/-1.08 vs. 1.69+/-0.47 and 2.28+/-0.73 pg/mL, p<0.05, respectively. Conclusions. 1) Sera from PE but not from nonpregnant women or normal pregnancies increase EC monolayer permeability. 2) Increased lipid peroxides and IL-8 are candidates altering EC barrier function. 3) Antioxidant SOD preserves increased EC monolayer permeability induced by PE serum, suggesting that EC oxidative stress may be associated with altered EC barrier function in preeclampsia.

Adult↗

Serum fatty acids as biomarkers of fat intake predict serum cholesterol concentrations in a population-based survey of New Zealand adolescents and adults.

BACKGROUND: The results of randomized controlled trials indicate that the amount and type of dietary fat are important predictors of serum cholesterol concentrations. However, the results of observational studies show weak or no association between dietary fat intake and serum cholesterol. Serum fatty acids are valid biomarkers of fat intake and may improve dietary estimates. OBJECTIVE: The objective was to ascertain whether serum fatty acids are associated with serum cholesterol concentrations in New Zealand adolescents and adults. DESIGN: The current study was a cross-sectional, national, population-based survey of 2793 New Zealanders aged > or =15 y who participated in the 1997 National Nutrition Survey. The fatty acid composition of serum cholesterol esters, phospholipids, and triacylglycerols was measured. RESULTS: A 1-SD increase in myristic acid (14:0) in serum cholesterol ester, phospholipids, and triacylglycerol corresponded with increases in serum cholesterol of 0.19, 0.13, and 0.10 mmol/L, respectively, after adjustment of the regression analysis for sex, age, body mass index, ethnicity, and smoking. The mean difference in cholesterol concentrations between persons in the highest and the lowest quintiles of serum cholesteryl-myristate was 0.48 mmol/L (P for trend < 0.001). A 1-SD increase in the proportion of linoleic acid (18:2n-6) in serum cholesterol ester, phospholipids, and triacylglycerol corresponded with decreases in serum cholesterol of 0.07, 0.07, and 0.05 mmol/L, respectively. The difference in mean serum cholesterol between the highest and lowest quintiles of cholesteryl-linoleate was 0.18 mmol/L (P for trend = 0.019). CONCLUSION: Saturated and polyunsaturated fat intakes, measured by using fatty acid biomarkers, are important predictors of serum cholesterol concentrations in New Zealand.

Adolescent↗

Relationship between fasting serum glucose, age, body mass index and serum 25 hydroxyvitamin D in postmenopausal women.

OBJECTIVE: Because it has been reported that vitamin D, given to mother or infant, can prevent type I diabetes in children, that diabetes is more common in adults with low serum vitamin D and that insulin secretion and action are related to vitamin D levels in healthy young adults we examined the relationship between serum vitamin D metabolites and fasting serum glucose in patients attending our outpatient clinics. DESIGN: Retrospective examination of convenience sample of postmenopausal women attending our osteoporosis clinics. PATIENTS: A total of 753 postmenopausal women attending a university hospital outpatient clinic and not on any treatment known to affect glucose metabolism. MEASUREMENTS: Body weight and height, serum 25-hydroxyvitamin D [25(OH)D], serum 1,25-dihydroxyvitamin D [1,25(OH)2D], serum PTH and fasting serum glucose. RESULTS: On simple correlation fasting serum glucose was a positive function of age (P < 0.05), weight (P < 0.001) and body mass index (BMI) (P < 0.001) and a negative function of serum 25(OH)D (P < 0.001), but it was not significantly related to either serum 1,25(OH)2D, PTH or creatinine. When fasting serum glucose was regressed simultaneously on age, BMI and 25(OH)D, glucose was still an inverse function of 25(OH)D (P = 0.006). CONCLUSIONS: Fasting serum glucose increased as 25(OH)D levels fell throughout the range of serum 25(OH)D measured but the greatest increase was observed in those with 25(OH)D below 40 nmol/l.

Adult↗

Effects of human serum on the growth and metabolism of Neisseria gonorrhoeae: an alternative view of serum.

Humans are the sole reservoir of Neisseria gonorrhoeae, an organism which undergoes a marked increase in metabolic rate after exposure to a low-molecular-weight, heat-stable component(s) of human serum. Further studies on the effect of serum on gonococcal metabolism were undertaken. Gonococcal broth (GCB) is commonly used for in vitro cultivation of gonococci. Gonococci suspended in GCB plus 10% serum exhibited oxygen consumption rates of 139% (P less than 0.01) and 456% (P less than 0.01) of those suspended in GCB or Hanks balanced salt solution, respectively. A twofold increase in growth rate also resulted from the addition of 10% serum to GCB. Gonococcal 14C-labeled adenine incorporation increased threefold with 10% serum supplementation of Hanks balanced salt solution. Dialysis of serum in 1,000-molecular-weight exclusion tubing removed the stimulatory factor(s). Neither correction of anion-cation concentrations altered by dialysis nor addition of substances of known importance to the metabolism of gonococci (i.e., lactate, pyruvate, cysteine, ATP, AMP, NADPH, amino acids, malate, and glutathione) to dialyzed serum reconstituted stimulatory capacity. The effect of serum on gonococcal glucose-catabolic pathways was measured by modified radiospirometry. An apparent threefold increase in Entner-Doudoroff and pentose phosphate pathway activities was induced by 10% serum, as was the increased shunting of glucose-derived glyceraldehyde-3-phosphate into these pathways. These metabolic changes did not allow specific identification of the serum stimulatory factor(s). Acetate, the major by-product of gonococcal glucose catabolism, inhibited gonococcal oxygen consumption as previously reported. A high-molecular-weight serum component, probably albumin, reversed acetate-mediated inhibition of gonococcal oxygen consumption, identifying a second mechanism by which serum increases gonococcal metabolism. These results suggest that supplementation of growth media with serum should be considered to provide N. gonorrhoeae with conditions more consistent with its normal environment.

Blood Physiological Phenomena↗

High serum sex hormone-binding globulin (SHBG) and low serum non-SHBG-bound testosterone in boys with idiopathic hypopituitarism: effect of recombinant human growth hormone treatment.

We measured serum sex hormone-binding globulin (SHBG), total testosterone (T), non-SHBG-bound T, albumin-bound T, free T, and SHBG-bound T in 19 prepubertal boys with hypopituitarism. Serum SHBG decreased with age with a slope similar to that in 91 normal prepubertal boys at higher level, and therefore, it reached similar values at a later age. Serum SHBG was significantly higher in hypopituitary prepubertal boys [mean, 123 +/- 12 (+/- SE) nmol/L] than in normal prepubertal boys (76 +/- 4; P less than 0.001) despite the fact that their mean age was also higher (10.0 +/- 4 vs. 7.1 +/- 4.1 yr; P less than 0.001). In 4 boys with isolated hypogonadotropic hypogonadism (Kallman's syndrome), aged 15.6 +/- 1.5 yr, serum SHBG was 21 +/- 14 nmol/L, a value below the 95% confidence limit of the regression line in GH-deficient boys. The affinity constants of association of the SHBG-DHT complex were similar in hypopituitary and normal boys. Eleven of the 19 hypopituitary boys (mean chronological age, 8.3 +/- 2.5 yr; mean bone age, 4.1 +/- 2.1 yr) were treated with recombinant hGH (0.5 U/kg BW.week) for 1 yr. Their mean serum SHBG level before treatment was 154 +/- 14 nmol/L, and it decreased gradually to 106 +/- 5 nmol/L (P less than 0.01) after 12 months of treatment. The tendency toward normalization of serum SHBG during treatment suggested that GH deficiency was responsible for the high serum SHBG levels. Serum SHBG correlated negatively with age in both treated hypopituitary and normal boys, but the slope of the regression line was significantly steeper in treated hypopituitary boys (P less than 0.01). On the other hand, the mean serum non-SHBG-bound T level was 0.10 +/- 0.02 (+/- SE) nmol/L in hypopituitary boys, significantly lower than that in normal boys (0.21 +/- 0.02 nmol/L; P less than 0.02). Since serum total T concentrations were similar in the two groups, the higher serum SHBG concentration resulted in lower serum bioavailable T levels in the hypopituitary boys. These changes might explain the poor response to T treatment reported in GH-deficient patients. The lower serum non-SHBG-bound T concentrations in the GH-deficient boys suggest there may be delayed exposure of central nervous system structures to increased levels of sex hormones, which, in turn, may slow body maturation. This mechanism might play a role in the delay of puberty that occurs in patients with isolated GH deficiency.

Adolescent↗

Serum thyroglobulin autoantibodies: prevalence, influence on serum thyroglobulin measurement, and prognostic significance in patients with differentiated thyroid carcinoma.

The prevalence of circulating thyroid autoantibodies (TgAb or antithyroid peroxidase) was increased nearly 3-fold in patients with differentiated thyroid cancers (DTC) compared with the general population (40% vs. 14%, respectively). Serum TgAb (with or without antithyroid peroxidase) was present in 25% of DTC patients and 10% of the general population. Serial postsurgical serum TgAb and serum Tg patterns correlated with the presence or absence of disease. Measurements of serum Tg were made in 87 TgAb-positive sera by a RIA and two immunometric assay (IMA) methods to study TgAb interference. TgAb interference, defined as a significant intermethod discordance (>41.7% coefficient of variation) between the Tg RIA and Tg IMA values relative to TgAb-negative sera, was found in 69% of the TgAb-positive sera. TgAb interference was characterized by higher Tg RIA vs. IMA values and was, in general, more frequent and severe in sera containing high TgAb concentrations. However, some sera displayed marked interference when serum TgAb was low (1-2 IU/mL), whereas other sera with very high TgAb values (>1000 IU/mL) displayed no interference. An agglutination method was found to be too insensitive to detect low TgAb concentrations (1-10 IU/mL) causing interference. Exogenous Tg recovery tests were an unreliable means for detecting TgAb interference. Specifically, the exogenous Tg recovered varied with the type and amount of Tg added and the duration of incubation employed. Further, recoveries of more than 80% were found for some sera displaying gross serum RIA/IMA discordances. The measurement of serum Tg in DTC patients with circulating TgAb is currently problematic. It is important to use a Tg method that provides measurements that are concordant with tumor status. IMA methods are prone to underestimate serum when TgAb is present, increasing the risk that persistent or metastatic DTC will be missed. The RIA method used in this study provided more clinically appropriate serum Tg values in the group of TgAb-positive patients with metastatic DTC. Furthermore, as serial serum TgAb measurements paralleled serial serum Tg RIA measurements, TgAb concentrations may be an additional clinically useful tumor marker parameter for following TgAb-positive patients. Disparities between serial serum Tg and TgAb measurements might alert the physician to the possibility of TgAb interference with the serum Tg measurement and prompt a more cautious use of such data for clinical decision-making.

Adolescent↗

Serum lipids, acute phase proteins and serum cholinesterase in normal subjects.

Although serum cholinesterase (CHE) is elevated in some hyperlipidaemic subjects, the relationship between serum CHE and lipids in normolipidaemic subjects is scanty. Furthermore, serum CHE is reduced in conditions in which there is an acute phase response. Serum CHE activity was measured in 46 normal individuals (22 males and 24 females). There was no significant difference between the activity of serum CHE in males or females being 6.2 +/- 1.8 U1(-1) vs. 6.4 +/- 1.5 U1(-1) respectively (mean +/- SD). There was, however, a significant correlation between serum CHE and subject age (Spearman rho 0.35, p < 0.05). There was also a significant correlation between serum CHE and serum nonfasting triglyceride concentration (rho 0.34, p < 0.05) and also apolipoprotein B (rho 0.38, p < 0.05) but not serum cholesterol or HDL-cholesterol. Five serum acute phase proteins were measured namely serum alpha-1 antichymotrypsin (ACT), alpha-1-acid-glycoprotein (AGP), alpha-2-macroglobulin (AMG), C-reactive protein (CRP), haptoglobin (HAP). Only serum AGP showed a significant negative correlation with serum CHE (rho - 0.43, p < 0.02).

Acute-Phase Proteins↗

The effect of a test meal on serum group I pepsinogens (PG I) and serum gastrin in persons with normal gastric H+ secretion and in persons with achlorhydria.

Thirteen persons with normal pentagastrin-stimulated gastric H+ secretion and 17 with achlorhydria were studied with a liquid test meal after an overnight fast. Blood was drawn before and every 30 min for 180 min after start of the meal. Serum gastrin, serum PG I, and serum vitamin B12 were determined by radioassay methods. Serum PG I was significantly lower in the achlorhydric subjects than in the normal secretors. The meal induced a slight and late rise in serum PG I in the control group. In contrast, the meal caused a slight fall in the achlorhydric persons. Basal serum gastrin was significantly higher in the achlorhydric group, in whom the meal also caused a significant fall in serum gastrin, which contrasts sharply with the rise in the control group. Although serum gastrin fell significantly in the achlorhydric group, a meal-induced rise in serum gastrin was observed in some of the achlorhydric subjects with basal serum gastrin below 100 pmol/l. Serum vitamin B12 was reduced in 8 of the 17 persons with achlorhydria, and in these 8 subjects serum PG I was significantly lower than in those with achlorhydria and normal serum vitamin B12.

Achlorhydria↗

Parental history of allergy, maternal serum IgE & cord serum IgE.

BACKGROUND: Allergic reactions are responsible for much illness, irritation and disquiet in the life of the child. Recognition and management of allergy in childhood is of great importance to prevent major allergic syndromes. AIM: To correlate cord serum IgE with parental history of allergy and maternal serum IgE. SETTINGS AND DESIGN: In consultation with the attending gynecologist a survey of 950 pregnant women from different parts of Mumbai, visiting gynecology clinics was carried out. METHOD: 100 pregnant women were randomly recruited for the study. Allergic history of both the parents was collected. Blood samples of mothers and matched cord blood samples were collected. Total serum IgE levels were determined using solid-phase immunoradiometric assay. Total serum IgA from each cord blood sample was checked. OBSERVATION & RESULTS: (1) Positive correlation was observed between cord serum IgE and matched maternal serum IgE, with mothers having history of allergy. (2) There was no relation (P=0.99) between cord serum IgE and matched maternal serum IgE, where mothers had no history of allergy. (3) A significant elevation in cord serum IgE was found only with maternal but not with paternal history of allergy. (4) No elevation was found in cord serum IgE (0.5 IU/mL). (6) All cord serum samples had IgA< 0.06 IU/mL ruling out the possibility of contamination with maternal blood. CONCLUSION: The maternal history of allergic disease is associated with an elevated cord serum IgE among newborns. Cord serum IgE is a simple, non-invasive and convenient test, which can supplement the maternal history to provide more discriminative information about the allergic status of the newborn.

Female↗

Serum supplements and serum-free media: applicability for microcarrier culture of animal cells.

Several different combinations of serum supplements and also serum-free media were examined for their ability to support the growth of MRC-5 and Vero cells on Cytodex microcarriers. Greater economy of serum was achieved for routine microcarrier culture by selecting the type of serum supplement, on the basis of whether the supplement was to support attachment and growth of cells at low densities or growth of cells at later stages in the culture cycle. Further economy was achieved by altering the serum concentration according to the requirements of each stage of the culture cycle. More consistent results were obtained when batches of serum were selected on the basis of quality control tests with microcarrier culture as well as with monolayer culture. Low serum and serum-free media were able to support the growth of cells in microcarrier cultures. A mixture of DME/Medium 199 (50/50) containing foetal calf serum (0.5%, v/v), BSA and EGF supported growth of MRC-5 and Vero cells to nearly the same extent as DME/Medium 199 containing 10% (v/v) foetal calf serum. Serum-free media was used to achieve cell yields at least one half of those attained with serum-supplemented media. Maximum growth was obtained when DME/Medium 199 was supplemented with fibronectin, BSA, insulin, transferrin, putrescine, fetuin and EFG. Fetuin was omitted and replaced by dexamethasone and trace metals with only a slight reduction in cell yield. Fibronectin was essential in all serum-free media.

Animals↗

Noninvasive monitoring using serum amyloid A and serum neopterin in cardiac transplantation.

The monitoring of allograft function for cardiac transplant patients still relies on endomyocardial routine biopsies. We investigated the diagnostic value of noninvasive monitoring using the parameters serum amyloid A protein and serum neopterin. The circulating levels of the acute phase reactant, amyloid A protein, and the macrophage product, neopterin, were measured serially in 13 patients after cardiac transplantation. The mean period of observation was 240 days. Nine acute cardiac allograft rejections, five cases of viral infection and eight cases of bacterial infection occurred. The levels of serum amyloid A protein and serum neopterin remained low (x = 6.0 mg/dL and 12.6 nmol/L, respectively) during the periods of stable graft function. In contrast, both parameters were significantly elevated (p < 0.01) during the rejection episodes (x = 12.7 mg/dL and 38.0 nmol/L for serum amyloid A protein and serum neopterin, respectively). For a reliable differentiation between rejection and stable graft function, serum amyloid A protein had a diagnostic accuracy of 84% (with a cut-off level of 10 mg/dL) and serum neopterin had one of 75% (with a cut-off level of 23 nmol/L). However, significant increases in the circulating levels of serum amyloid A protein and serum neopterin were also observed during bacterial (x = 14.9 and 88 nmol/L, respectively) and viral (x = 6.2 mg/dL and 44 nmol/L, respectively) infections. The detection of immunological complications after cardiac transplantation using serial measurements of serum amyloid A protein and serum neopterin is possible. These parameters can be used to help in judging both the need and the optimal timing for the otherwise frequent endomyocardial biopsies.

Antilymphocyte Serum↗