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Query: UMLS:C0432222 (SEM)
 47,337 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The technique originally described by Kumon et al. for the labeling of cell surface antigens under the SEM has been adapted to the study of surface immunoglobulins on human lymphocytes. Briefly, the peripheral blood lymphocytes are: 1) separated in a Ficoll Hypaque gradient, 2) rinsed with culture medium, 3) allowed to attach to plastic coverslips, 4) prefixed with 1% glutaraldehyde for 10 minutes, 5) incubated with goat anti-human immunoglobulins for 30 minutes at 24 degrees C, 6) rinsed, 7) incubated with a rabbit anti-goat IgG coupled with purified bacteriophage T4, 8) rinsed, 9) postfixed in glutaraldehyde for several hours, and 10) dried from ethanol at -75 degrees C and under 10(-2) Torr. Results from normal human lymphocytes and cells from cases of Chronic Lymphoblastic Leukemia (CLL) indicate that the method makes it possible to recognize cells with surface immunoglobulins and permits some correlation between the surface architecture of the cells and the presence of surface Ig. It also permits the study under the SEM of T-derived lymphocytes and of null cells, the unlabeled cell population, without resorting to techniques for lymphocyte subclass separation which may alter surface morphology.
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PMID:Labeling of lymphocyte surface immunoglobulins with T4 as marker for scanning electron microscopy. 52 28

Previous studies have shown an abnormality of the spermidine-to-spermine (Spd/Spm) ratio in whole blood of cystic fibrosis homo-and heterozygotes. To investigate Spd and Spm distribution amoung blood components as a possible cause of the abnormality, blood was fractionated using Rabinowitz's glass bead technique and Boyum's Ficoll-Hypaque method. Free (unconjugated) polyamines were extracted with perchloric acid and quantitated on an amino acid analyzer. In controls, mean +/- SEM concentrations in nmoles/10(9) cells of Spd and Spm, respectively, were 1.02 +/- 0.08 and 0.894 +/- 0.28 for erythrocytes; 126 +/- 31 and 357 +/- 105 for lymphocytes; 36 +/- 16 and 240 +/- 33 for granulocytes; and less than 0.5 and less than 0.5 nmoles/ml for plasma. When converted to the concentration in whole blood, it was found that greater than 90% of Spd and over 70% of Spm was associated with erythrocytes. While the higher cellular concentration in leukocytes was not unexpected, the fact that Spd and Spm in whole blood were primarily associated with erythrocytes was a new finding. Comparison with controls revealed that the Spd/Spm ratio in both whole blood and erythrocytes was significantly higher in the group of cystic fibrosis patients.
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PMID:Distribution of spermidine and spermine in blood from cystic fibrosis patients and control subjects. 95 66

The relationship between age and blood magnesium (Mg) parameters has not been defined. Mg measurements in plasma, red blood cells (RBCs), and mononuclear blood cells (MBCs) were made in 104 normal volunteers (43 males and 61 females, ages 11-75 years). MBCs were separated from blood using a discontinuous Ficoll-Hypaque gradient. The mean values (+/- SEM) were as follows: plasma Mg 1.63 +/- 0.01 mEq/L, RBC Mg 4.55 +/- 0.06 mEq/L, MBC Mg content 72.8 +/- 1.0 fg/cell, and MBC Mg concentration 19.6 +/- 0.3 mEq/L. We compared these parameters with age (intervals of 10 years) using analysis of variance (ANOVA) and found no significant differences (p greater than 0.05). Thus, plasma, RBC, and MBC Mg parameters do not vary significantly between the ages of 11 and 75 years.
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PMID:Blood magnesium parameters do not differ with age. 221 87

Effects of ionic (Hypaque-76) and nonionic (Isovue-370 and Omnipaque-350) contrast media on oxyhemoglobin dissociation of normal human red blood cells were evaluated. In series 1, 4-mL venous blood samples were obtained from 15 normal human volunteers. One blood sample served as control, and 1 mL of either of the three contrast media was added in vitro to the other 4-mL blood samples. P50 values were estimated from the linear portion of the oxyhemoglobin dissociation curve obtained by tonometry. Determinations of P50 were performed at either pH 7.4 or 7.2. At pH 7.4, P50 in the absence of contrast media was 26.3 +/- 0.4 mm Hg (mean +/- SEM). The contrast media caused comparable decreases in P50 from this value (Hypaque-76, 20.0 +/- 0.5 mm Hg; Omnipaque-350, 21.6 +/- 0.4 mm Hg; Isovue-370, 20.7 +/- 0.4 mm Hg). Reducing pH to 7.2 in the absence of contrast media increased P50 to 33.3 +/- 1.0 mm Hg, evidence of the Bohr effect. The presence of contrast media either completely abolished (Hypaque-76 and Omnipaque-350) or markedly attenuated (Isovue-370) this effect. In series 2 (five patients), blood samples were withdrawn from the external iliac artery during injection of Isovue-370 (60-78 mL) into the proximal abdominal aorta to evaluate peripheral vascular disease. Measurement of P50 of these samples yielded findings consistent with those of series 1. The present findings demonstrate that both ionic and nonionic contrast media increase the affinity of hemoglobin for oxygen and, therefore, that they may inhibit oxygen delivery to body tissues.
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PMID:Contrast media adversely affect oxyhemoglobin dissociation. 236 33

Bone marrow samples from normal adults and children with nonhematologic malignancies not involving the marrow, acute lymphoblastic leukemia (ALL) in continued remission, and immune cytopenias were studied by two-color immunofluorescence (IF) and flow cytometry to characterize common acute lymphoblastic leukemia antigen (CALLA)-positive marrow lymphoid cells. Marrow was separated by Ficoll/Hypaque centrifugation followed by passage over a monoclonal antibody affinity column to remove myeloid cells prior to IF staining. A higher proportion of CALLA-positive cells was found in the pediatric marrows (mean, 33.7% +/- 6.3% SEM) than in the adult marrows (mean, 4.5% +/- 1.6% SEM). Two subpopulations of CALLA-positive cells identified by cell sorting to be of lymphoid morphology were found in both adult and pediatric marrows. A small subpopulation comprising 12.3% of the total CALLA-positive cells was characterized by a high intensity of CALLA, terminal deoxynucleotidyl transferase (TdT), and MY10 expression, but low B1, common leukocyte antigen, and peanut agglutinin receptor expression. The remainder of the CALLA-positive cells displayed low intensity of CALLA expression, positivity for the common leukocyte antigen, B1 and peanut agglutinin, and negative reaction with TdT and MY10. Both CALLA-positive subpopulations were positive for HLA-DR and the pan-B cell marker B4, but negative for cytoplasmic immunoglobulin, B2 and Leu 1. BrdU labelling studies showed that a similar proportion of cells in each subpopulation was in S phase. A slightly higher proportion of the strongly CALLA-positive cells possessed the morphologic features of high nuclear/cytoplasmic (N/C) ratio and prominent nucleoli. These studies suggest that a discrete maturation step occurs among CALLA-positive marrow lymphoid cells, resulting in the loss of TdT and MY10 expression, but gradual acquisition of the B cell marker B1 and the common leukocyte antigen. The presence of B4 antigen in nearly all CALLA-positive cells suggests that both subpopulations of normal CALLA-positive marrow cells are committed to the B cell lineage.
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PMID:Subpopulations of common acute lymphoblastic leukemia antigen-positive lymphoid cells in normal bone marrow identified by hematopoietic differentiation antigens. 294 98

To characterize the insulin-like growth factor I (IGF-I) receptor on human erythrocytes, cells were purified from peripheral blood by Ficoll-Hypaque gradient centrifugation and incubated with [125I]IGF-I. Specific binding was maximal at pH 8.0 after 24 h at 4 C and increased linearly with cell number to 3.9 +/- 0.2% (+/- SEM) for 3.0 X 10(9) cells/ml. The Scatchard plot of the binding data was linear, with 7 fmol [125I]IGF-I bound/10(9) cells and an affinity constant (K) of 1.8 X 10(9) M-1. Unlabeled IGF-I inhibited tracer binding half-maximally at 6 ng/ml. Multiplication-stimulating activity (or rat IGF-II) was 40% as potent (ED50, 15 ng/ml), whereas insulin and proinsulin were 30- to 500-fold less potent. A monoclonal antibody to the IGF-I receptor (alpha IR-3) inhibited IGF-I binding by 50% at a 1:1000 dilution and by 80% at a 1:250 dilution. Insulin binding was unaffected by the same dilutions. IGF-I receptor phosphorylation was studied in erythrocyte ghosts prepared by hypotonic lysis and solubilized in 1% Triton. The extract was preincubated with and without 100 ng/ml IGF-I or porcine insulin and incubated with [gamma-32P]ATP in the presence of Mn2+, and the receptor was identified by immunoprecipitation with alpha IR-3 antibody and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. IGF-I stimulated 4-fold the incorporation of 32P into a protein of 95,000 mol wt, which was immunoprecipitated by alpha IR-3; insulin produced a 2-fold stimulation of this protein. This protein corresponds to the beta-subunit of the IGF-I receptor. These data demonstrate that human erythrocytes have specific receptors for IGF-I, and that this IGF-I receptor, like the insulin receptor, undergoes ligand-stimulated autophosphorylation. Thus, analysis of erythrocyte IGF-I binding and receptor phosphorylation may be useful tools for the study of patients with a variety of growth disorders.
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PMID:The human erythrocyte insulin-like growth factor I receptor: characterization and demonstration of ligand-stimulated autophosphorylation. 300 55

Multiple intravenous injections of contrast media are used in radiology, but information regarding corresponding changes in the serum iodine concentrations and osmolality is lacking. We measured the changes in serum iodine concentration and serum osmolality in rabbits after a series of intravenous injections of contrast media. Hypaque-76 (1 cc/kg) was injected intravenously in awake rabbits at 10-minute intervals for 1 hour and arterial blood sampled at midpoint times between injections. During the 1-hour period of seven serial injections, mean serum iodine concentration at 5 minutes was 2.3 mg I/cc (+/- 0.1 SEM, n = 14); at 35 minutes, 5.1 mg I/cc (+/- 0.2); and at 65 minutes, 6.7 mg I/cc (+/- 0.4). The mean peak concentration was 6.8 mg I/cc (+/- 0.4). Serum osmolality underwent a mean increase of 16 mosm/kg during the injection period. Hypertonic mannitol injections produced a smaller increase in osmolality (10 mosm/kg). Isotonic saline injections in control animals produced no change in osmolality.
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PMID:Effect of multiple intravenous injections of diatrizoate on iodine concentrations and osmolality. An experimental study in rabbits. 310 30

Macrophages have been identified in the developing corpus luteum in several species, including man, and also constitute approximately 90% of cells in the peritoneal cavity. We studied the effect of peritoneal macrophages or blood monocytes on progesterone (P) synthesis by human granulosa cells from preovulatory follicles obtained at laparoscopy of 14 women undergoing in vitro fertilization. Pooled granulosa cells from follicles with mature ova were isolated by Ficoll-Hypaque gradient centrifugation. Washed granulosa cells (0.75 X 10(5)/ml) were incubated in Dulbecco's Minimum Essential Medium containing 20% calf serum with varying concentrations of pelvic macrophages (0.8-29 X 10(5)/ml) or fresh and mature blood monocytes (0.25-2.5 X 10(5)/ml). P production was determined by RIA of medium at 24-h intervals for 24-48 h. In situ concentrations of pelvic macrophages from 8 patients with tubal infertility increased cumulative P production to 140 +/- 17.8% (mean +/- SEM) of the control values. A similar increase (182 +/- 62.7%) was found with macrophages from 6 patients with endometriosis or unexplained infertility. Both fresh and mature monocytes stimulated P production to 225% and 261% of control values, respectively. Indomethacin (10(-4) M) or monoclonal antibody to somatomedin-C did not prevent stimulation of P production. These results suggest that peritoneal macrophages may exert luteotropic effects on cumulus cells while the ovulated oocyte resides in the tube, and incoming monocytes may be important in stimulating luteal cells in the developing corpus luteum.
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PMID:Peritoneal macrophages modulate human granulosa-luteal cell progesterone production. 404 79

Investigation of the pathogenesis of inflammatory diseases has involved assessment of polymorphonuclear leukocyte (PMN) activity and a variety of techniques for separating PMNs from whole blood have been described. In this study the effects of Percoll gradient, Ficoll-Hypaque/Dextran sedimentation (FH/DS) and Mono-Poly Resolving Medium (M-PRM) on activation and function of PMNs were compared. All three separation techniques gave similar cell yield and purity. The mean (+/- SEM) percentage of cells demonstrating pseudopodia formation for Percoll, FH/DS and M-PRM were 39 +/- 9, 57 +/- 6 and 63 +/- 5, respectively, while superoxide release from resting cells was 1.9 +/- 0.9, 7.2 +/- 3.5 and 11 +/- 4.8 pmols per 10(6) cells min-1, respectively, indicating that activation of cells during separation may be less with Percoll compared to the other methods. The functional capacity of the cells to respond to a stimulus was similar for all methods as indicated by similar EC50 values for chemotaxis to zymosan-activated serum and similar superoxide production induced by tetradecanoyl phorbol acetate. All three separation techniques produce functionally active PMNs of high purity but the use of Percoll gradients may be preferable when a quick method of separation which causes minimum pre-activation of PMNs is required.
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PMID:Effects of different density gradient separation techniques on neutrophil function. 752 31

The aim of this study was to prospectively analyze the possible association of delayed gastric emptying and postoperative pancreatic complications after pancreaticoduodenectomy. Although hospital mortality after pancreaticoduodenectomy is minimal, morbidity is still high; delayed gastric emptying is one of the most frequent complications. Thirty-nine consecutive patients undergoing pancreaticoduodenectomy were included in this study: 14 females and 25 males (median age 65 years; range, 7-82). Delayed gastric emptying was defined as the need for a nasogastric tube or recurrent vomiting that prevented normal feeding on the 10th postoperative day. Blood analysis was performed on postoperative days 4, 6, and 10; Gastrografin examination on day 6; CT scan on days 2 and 5; and drain amylases were measured on day 5. Pancreatitis was defined as pancreatitis changes in CT scan interpreted by an experienced radiologist without knowing other data. Pancreatic fistula was defined according to the recent international recommendations. We had no mortality. Twelve patients (31%) developed delayed gastric emptying. Surgical (9/12 vs. 5/27; P = 0.001) but not medical complications occurred more often in the delayed gastric emptying group. Of the single complications, postoperative CT-detected pancreatitis (6/12 vs. 4/27; P = 0.03) and postoperative pancreatic fistula (5/12 vs. 1/27; P = 0.0007) were significantly associated with delayed gastric emptying compared with the patients without delayed gastric emptying. This pancreatitis was already detected in CT scan on day 2 in most patients (6/10, 60%). In delayed gastric emptying patients, the only parameters in blood analysis that differed significantly from patients without this complication were serum amylase activity (mean +/- SEM, 715 +/- 205 vs. 152 +/- 70 IU/L; P = 0.02), blood leukocyte count (16 +/- 2 vs. 9 +/- 0.6 x 10(9)/L; P = 0.007) and serum C-reactive protein (CRP) concentration (144 +/- 28 vs. 51 +/- 14 mg/L, P = 0.01). Postoperative pancreatic (subclinical) fistula was also associated with postoperative pancreatitis (6/10 vs. 0/29; P = 0.003). Preoperative coronary artery disease (OR = 16; 95% CI, 1.0-241; P = 0.05) and soft pancreatic texture at operation (OR = 9; 95% CI, 1.4-52; P = 0.02) were significant risk factors for the development of postoperative pancreatitis. The diagnosis of delayed gastric emptying after pancreaticoduodenectomy often follows postoperative pancreatitis. Delayed gastric emptying is also associated with postoperative pancreatic fistula, for which this pancreatitis seems to be a risk factor. Preoperative coronary artery disease and soft texture of the pancreas are significant risk factors for postoperative CT-detected pancreatitis.
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PMID:Postoperative acute pancreatitis as a major determinant of postoperative delayed gastric emptying after pancreaticoduodenectomy. 1696 32


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