Showing papers on "Albumin published in 1982"

Selective Transport of Polymeric Immunoglobulin-a in Bile - Quantitative Relationships of Monomeric and Polymeric Immunoglobulin-a, Immunoglobulin-m, and Other Proteins in Serum, Bile, and Saliva

Abstract: In 17 adults, serum, hepatic bile, and saliva samples were analyzed for their sedimentation profile of IgA and secretory component (SC), and for their concentrations of albumin, orosomucoid, transferrin, IgG, IgA, alpha 2-macroglobulin (alpha 2M), IgM, and SC. Polymeric IgA(p-IgA) averaged 13% (50-700 micrograms/ml) of total IgA in serum, 70% (43-88%) in bile, and 93% (74-98%) in saliva. Most of the p-IgA in bile sedimented with SC, which also occurred free (8-44%), and with IgM. In bile, albumin (155-1,485 micrograms/ml) was the predominant protein, followed by IgG (32-480 micrograms/ml), and total IgA (37-209 micrograms/ml). In saliva, p-IgA (72-902 micrograms/ml) predominated, followed by albumin (16-385 micrograms/ml) and IgG (9-178 micrograms/ml). Secretion-to-serum albumin-relative concentration ratios (S/S-ARCR = 1 for albumin) in bile averaged 22 for p-IgA, 1.91 for IgM, 1.28 for monomeric IgA (m-IgA), 0.70 for IgG, and 0.57 for alpha 2M, indicating for p-IgA, IgM, and to a lesser extent for m-IgA, a selective excretion into bile. In saliva, a 16-fold greater selective excretion of p-IgA (mean S/S-ARCR = 354) was found. Labeled m- and p-IgA were injected intravenously into five patients. Specific activities indicated that for p-IgA 50% was serum derived in bile, as compared with 2% in saliva, and to 85% for m-IgA in bile. In the patient with the highest excretion of 125I-p-IgA in bile, only 2.8% of the injected dose was recovered in bile within 24 h after injection. Compared with rats and rabbits, the serum-to-bile transport of p-IgA in humans is much smaller.

Isolation and preliminary characterization of a fraction of bilirubin in serum that is firmly bound to protein.

Abstract: We have isolated from pathological sera a bilirubin fraction (delta) that is very tightly, if not covalently, bound to protein, most likely albumin. This delta fraction absorbed at a lambda max of 433 nm in the visible spectrum, between the lambda max of unconjugated (alpha) and that of conjugated (Bc) bilirubin when measured in solutions containing albumin. However, unlike the other bilirubin species, this fraction could not be separated from the proteins in serum by exhaustive ultrafiltration in the presence of caffeine/benzoate solution. In the Jendrassik-Grof diazo procedure for bilirubin analysis, the delta fraction gave a large direct reaction (76-89% of the total reaction). Yet, when relatively hydrophobic azo dyes were formed by reaction of the delta fraction with the diazonium salt of dichloroaniline, only 50% of the dyes were extractable from aqueous solution. On chromatography the rest remained associated with protein. Of the extractable dye, more than 70% was accounted for by two liquid-chromatographic peaks with retentions identical with those of azo dyes formed from unconjugated bilirubin. This delta fraction was not appreciably separated from protein by treatment with strong acid or base, or by prolonged digestion with various enzymes. Finally, in a highly denaturing solvent (urea/mercaptoethanol), this fraction was not dialyzable through a membrane with a 12 000-dalton cutoff.

Human tears: Normal protein pattern and individual protein determinations in adults

Abstract: Proteins in tears are known to be highly heterogeneous. We have tried to take into consideration some of the difficulties encountered in studying tear proteins such as tear collection, the choice of the parameters to be studied, the adequacy of techniques for small sample volumes, etc. Levels of total tear proteins were determined in 101 samples and the electrophoreses were performed. On gel slabs, four major proteins were always found to be present : lactoferrin, lysozyme and two tear specific proteins. Concentrations of some individual proteins, lactoferrin, lysozyme, IgA, IgG and albumin were determined. Comparisons of lactoferrin and lysozyme contents were found to be more reliable when based on total protein rather than on their own concentrations as such.

Effects of ischemia and oxygen radicals on mucosal albumin clearance in intestine

Abstract: Mucosal albumin clearance was measured in jejunal segments of dogs under control conditions, after arterial occlusions of varying duration (15 min-4 h), and during intraluminal perfusion with hypoxanthine-xanthine oxidase. Albumin clearance rates were estimated from the luminal perfusion rate and the activity of protein-bound 125I in the perfusate and plasma. Arterial occlusion of 30-min to 4-h duration produced a significant increase in mucosal albumin clearance. The magnitude of the rise in albumin clearance was directly related to the duration of arterial occlusion. Pretreatment with superoxide dismutase, a superoxide radical scavenger, or allopurinol, a xanthine oxidase inhibitor, did not prevent the increased albumin clearance induced by 1 h of occlusion. Intraluminal perfusion with hypoxanthine-xanthine oxidase significantly increased mucosal albumin clearances. This increase was prevented by superoxide dismutase. The results of this study indicate that arterial occlusions and enzymatically generated superoxide radicals increase mucosal albumin clearance.

Urinary excretion of albumin in normal man: The effect of water loading

Abstract: The urinary excretion of albumin and (β2-microglobulin in response to two different types of oral water load was studied in 18 healthy subjects. Both acute water loading (1 litre of tap water given over 10 min) and chronic water loading (250 ml of water given every 20 min for the 4-h duration of the test) produced a short-lived but significant increase in the urinary excretion of albumin, but not in β2-microglobulin. Albumin excretion returned promptly to basal values in spite of high and sustained urine flow. The decrease in haematocrit, plasma osmolality, plasma sodium concentration and pulse rate and the increase in creatinine clearance suggest that the elevation in albumin excretion might be the result of a transient volume expansion associated with an increase in glomerular filtration rate and in filtration of albumin. However, a washout of proteins from the tubular lumen with preferential reabsorption of small molecular weight proteins (i.e. β2-microglobulin) during high tubular flow cannot be exclu...

The immunological identification of brain proteins on cellulose nitrate in human demyelinating disease.

Abstract: An immunological technique has been employed to identify proteins, separated in polyacrylamide gels, which show changes in brain samples from cases of multiple sclerosis and subacute sclerosing panencephalitis. Sodium dodecylsulphate-treated proteins in particulate and soluble fractions were separated in polyacrylamide slab gels, transferred electrophoretically onto cellulose nitrate sheets, incubated with specific antisera and visualized by an immunoperoxidase method. Protein bands showing changes were identified using antisera raised against the myelin basic and Wolfgram proteins, the neurofilament triplet proteins, tubulin and glial fibrillary acidic protein. In addition to the loss of myelin proteins, decreases in the neurofilament proteins and in tubulin were seen in both multiple sclerosis and subacute sclerosing panencephalitis samples. The distribution of glial fibrillary acidic protein polypeptides in the particulate and soluble fractions of plaque samples appeared to vary according to the degree of fibrosis. Changes in the levels of the myelin-associated glycoprotein, the lower molecular weight component of the Wolfgram protein, albumin and immunoglobulin G, none of which were visualized by protein staining, were also seen. This immunological technique has allowed a closer examination of changes occurring in brain protein spectra in multiple sclerosis and subacute sclerosing panencephalitis.

Proteins of human urine. III. Identification and two-dimensional electrophoretic map positions of some major urinary proteins.

Abstract: We mapped the proteins of human urine by high-resolution two-dimensional electrophoresis, utilizing the ISO-DALT system. Wide-range pH gradients and narrow-range acid gradients were both used in the first-dimension separations. The patterns revealed proteins ranging in relative molecular mass from 10 000 to 90 000. Proteins identified in the map included transferrin, albumin, hemopexin, alpha 2-HS glycoprotein, alpha 1-antitrypsin. Gc globulin, alpha 1-acid glycoprotein, Zn alpha 2-glycoprotein, retinol binding protein, beta 2-microglobulin, the immunoglobulin light chains, and MAUP (most acid urinary protein). The use and utility of internal-charge and molecular-mass standards are described. We used electrophoretic transfer of proteins to nitrocellulose sheets and subsequent detection by immunological methods to identify some proteins.

Mechanisms of proteinuria in human glomerulonephritis.

Abstract: We evaluated glomerular barrier function in 28 patients with glomerulonephritis. Neutral dextrans of graded size were used to characterize the size-selective properties of the barrier. Charge selectivity was characterized by electrofocusing excreted urinary proteins. A fractional IgG clearance (relative to freely permeable inulin), smaller or greater than 100 x 10(-5) was used to distinguish patients with minor (group I, n = 13) and major (group II, n = 15) urinary IgG leakage, respectively. Fractional clearances of smaller dextrans (radii 20-50 A) were similar, but those of larger dextrans (radii 52-60 A) were elevated in group II relative to group I patients. A model of solute transport through a bimodal pore size distribution revealed the values for pore radius in the lower mode to approximate 51-55 A in both group I and group II patients. Pore radius in the upper mode, by contrast, was much larger in group II than in group I patients, approximating 87-97 vs. 72-77 A, respectively. Electrofocusing of urinary protein from group I patients revealed mostly albumin (isoelectric point 5.2). In group II patients, however, immunoglobulin excretion was copious. Moreover, the distribution of anionic, neutral, and cationic species (isoelectric points 5.5-8.5) in urinary and plasma eluates of IgG2 and IgG4 was similar. We conclude that when glomerulonephritis is associated with selective albuminuria, as in group I,, there is an isolated reduction of electrostatic retardation of relatively small anionic proteins. Major urinary IgG leakage (group II), however, appears to result from the development in the glomerular membrane of a subpopulation of enlarged pores that are highly permeable towards proteins of large size and varying charge.

Relationship of serum total calcium to albumin and total protein in dogs.

Abstract: A positive linear relationship was found between total calcium and albumin and between total calcium and total protein in the serum of 209 dogs. Total calcium concentration correlated with the concentration of albumin (r = 0.575; P less than 0.001) and with the concentration of total protein (r = 0.411; P less than 0.001). A correction formula for calcium was derived on the basis of the concentration of albumin: adjusted calcium (mg/dl) = calcium (mg/dl) - albumin (g/dl) + 3.5. The correction formula for calcium, based on the concentration of serum total protein was: adjusted calcium (mg/dl) = calcium (mg/dl) - 0.4 [total serum protein (g/dl)] + 3.3. Hypocalcemia (less than or equal to 8.7 mg/dl) was detected in 32 of the dogs. After adjustment of the measured total calcium for albumin and serum total protein, 29 (91%) of the dogs had calcium concentrations within the normal range. Hypercalcemia was not associated with hyperalbuminemia or hyperproteinemia. In 91% of dogs with disorders of calcium metabolism and in 86% of dogs less than 6 months old, calcium concentrations were outside the 95% confidence intervals for albumin and total protein calculated from the 209 dogs. It was concluded that adjustment of serum total calcium for protein concentration is essential for correct interpretation of calcium values and detection of abnormalities in calcium metabolism.

Study of protein characteristics that influence entry into the cerebrospinal fluid of normal mice and mice with encephalitis.

Abstract: Entry of proteins into the cerebrospinal (CSF) from the blood is partially determined by the size of the protein. To determine whether other characteristics of proteins influence CSF entry, proteins or protein fragments were iodinated, inoculated intravenously, and serum and CSF were sampled at later times. The Fc fragment of immunoglobulin G (IgG) did not enter the CSF significantly better than the Fab fragment suggesting that choroidal Fc receptors are not of importance for selective immunoglobulin entry. To determine the role of protein charge on entry, bovine serum albumin [isoelectric point (pI) = 3.9] was chemically altered to provide an albumin with an average pI of 6 (A-6) and another with a pI of 8.5 (A-8). All albumins were of the same size on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A-8 entered the CSF approximately 10-fold better than the native albumin. A-6 was intermediate, entering approximately twofold better. At the time of increased CSF protein concentration during an acute viral encephalitis these differences were narrowed but not eliminated. It is concluded that charge is an important determinant of protein entry into the CSF.

Blood chemistry homeostasis during prolonged fasting in the northern elephant seal

Abstract: Serum electrolytes, 3 enzymes, and 11 metabolites were monitored for 32-68 days in weaned, naturally fasting elephant seal pups. Serum glucose, urea nitrogen and creatinine levels declined as the fast progressed, whereas total protein, albumin, and globulin levels remained nearly constant. By contrast, triglycerides, cholesterol, uric acid, and bilirubin were quite variable and no definite trends were apparent. Alkaline phosphatase activity appeared to increase during fasting, while serum glutamic oxalacetic transaminase and lactate dehydrogenase remained fairly uniform. Comparisons of averaged blood chemistry values from singly and multiply sampled fasting pups to that of four nursing pups showed significant differences in the levels of blood urea nitrogen, creatinine, cholesterol, bilirubin, and albumin, but sampling uncertainty limited physiological interpretation. Electrolyte levels in all animals were maintained within narrow limits under all conditions with little interindividual variation. These results further document the remarkable homeostasis achieved during prolonged fasting in elephant seals and support the hypothesis that fat is the primary energy substrate during the protracted natural fasts characteristic of this species.

Relationship between platelet aggregation, thromboxane synthesis and albumin concentration in nephrotic syndrome

Abstract: Summary. Increased platelet aggregability on stimulation with sodium arachidonate (NaAA), collagen and ADP was found in a group of 14 patients with nephrotic syndrome when compared with age and sex matched controls. Five of the group also exhibited spontaneous platelet aggregation (SPA), associated with synthesis of thromboxane B2 (TxB2), and which appeared to correlate with a markedly increased serum triglyceride concentration. Thromboxane B2 generation in response to NaAA was increased and reflected both the low serum albumin concentration and the increased platelet aggregation response to this agent. Addition of albumin in vitro decreased the amount of TxB2 generated for a given dose of NaAA and increased NaAA and collagen-induced platelet aggregation thresholds. However, albumin had no significant effect on collagen-induced TxB2 production. The results suggest that the hypoalbuminaemia and associated reduced binding of arachidonic acid and increased synthesis of TxA2 account in part for the increased platelet aggregability seen in the nephrotic syndrome but that other mechanisms are also involved.

Altered plasma drug binding in cancer: Role of α1‐acid glycoprotein and albumin

Abstract: Altered concentrations of serum proteins often accompany malignant disease. The effect of these changes on drug binding was studied with lidocaine, a basic drug, and tolbutamide, an acidic drug. Patients with cancer had increased serum concentrations of the acute-phase protein α1-acid glycoprotein (AAG) and lowered serum concentration of albumin. In association with these changes lidocaine binding was increased at all concentrations studied (predialysis concentrations 2, 6, and 10 μg · ml−1) and that of tolbutamide was decreased at the highest concentration (200 μg · ml−1). Not all of the increase in lidocaine binding was explicable on the basis of increased serum AAG concentration. Estimation of binding parameters with a model with two independent sites showed increased affinity at the high affinity site in cancer patients with no change in the calculated number of binding sites. Therefore, in cancer there is increased lidocaine binding in association with increased AAG concentrations. We also record the novel observation of a change in the intrinsic properties of the high affinity binding site. Clinical Pharmacology and Therapeutics (1982) 32, 295–302; doi:10.1038/clpt.1982.163

Nonenzymatic glycosylation of serum and plasma proteins

Abstract: Albumin and other serum and plasma proteins may be nonenzymatically glycosylated in vitro and in vivo. Chromatographie and colorimetrie measurement of these nonenzymatically glycosylated proteins shows that levels are approximately two to three times higher in diabetic than nondiabetic subjects. In diabetic patients levels appear to reflect glycemia in the preceding several days and correlate with other established parameters of glycemia such as fasting serum glucose, mean of several capillary blood glucose measurements, and glycosylated hemoglobin. Because serum and plasma proteins have shorter half-lives than hemoglobin, nonenzymatically glycosylated proteins are altered more rapidly than glycosylated hemoglobin in response to prolonged decreases or increases in blood glucose levels. Acetylsalicylic acid significantly inhibits in vitro glycosylation of albumin and other serum proteins incubated for up to 7 days in glucose. It is not known whether such inhibition occurs in vivo. Glycosylation of albumin appears to reduce its solubility, and glycosylation of low-density lipoprotein may increase its catabolism. Whether nonenzymatic glycosylation of other serum proteins alters their functions or may contribute to some of the long-term complications of diabetes is unknown at present.

Plasma protein concentrations in interstitial fluid from human aortas.

Abstract: The concentration of plasma proteins was examined in interstitial fluid collected from human aortas, obtained at autopsy, from patients from whom a blood sample had been taken for routine analysis shortly before death. The interstitial fluid was absorbed onto small, preweighed pieces of filter paper which were inserted into natural strip planes in the tunica intima and inner tunica media. After equilibration the papers were removed and weighed to measure the amount of interstitial fluid collected, then analysed by quantitative immunoelectrophoresis for three plasma proteins covering a range of molecular masses ( M r ): low density lipoprotein (LDL) with molecular mass 2.4 x 10 6 , α 2 -macroglobulin ( M r 720 000) and serum albumin ( M r 68 000). In interstitial fluid obtained from normal intima of 20 men and women, the mean levels of the proteins were LDL 215%, α 2 -macroglobulin 115% and albumin 54% of the concentration in the patient’s own plasma. Concentrations were not influenced by depth within the intima, or by distance down the aorta. The patients covered the age range 31-96 years; relative concentrations of LDL, α 2 -macroglobulin and albumin increased in parallel by about 10% per decade. Large samples (up to 7 μl) of interstitial fluid were collected from inner media but they contained no measurable LDL; the levels of α 2 -macroglobulin and albumin were respectively 11 and 18% of plasma concentration. Analysis of interstitial fluid from one sample of normal intima obtained at vascular surgery, and from two freshly killed pigs, suggested that results were not invalidated by the use of autopsy material. Direct comparison of whole intimal tissue and interstitial fluid provided no evidence of preferential binding of LDL in tissue. There was no evidence of preferential adsorbtion of LDL by the filter paper. Interstitial fluid of six samples of normal intima from four patients was compared with normal plasma by two-dimensional immunoelectrophoresis. The peaks produced in the antiserum to LDL were identical in mobility, shape and staining properties. Thus the concentration of LDL found in interstitial fluid from normal aortic intima was more than twice the plasma concentration, and the relation between concentration and molecular mass was the inverse of that reported for peripheral interstitial fluid and lymph. Endothelium appears to trap LDL in the intima instead of keeping it out, and this raises fundamental questions about the function of arterial endothelium.

Immunosuppressive effect of acute-phase reactant proteins in vitro and its relevance to cancer.

Abstract: Acute-phase reactant proteins reach abnormally high levels in patients with cancer, and correlate with the extent of disease. In this study, several acute-phase glycoproteins, and serum albumin as a control, were tested at different concentrations for their ability to modify the blastogenic response of lymphocytes from 30 normal donors to PHA and the chemotactic response of monocytes from 15 normal donors to casein. In high concentrations approximating those found in cancer patients, but not in normal concentrations, haptoglobin and fibrinogen inhibited both functions to different degrees. Orosomucoid inhibited only monocyte chemotaxis, while ceruloplasmin and α1-antitrypsin affected neither function.

Immune reactions in the lungs of asymptomatic dairy farmers.

Abstract: To test the hypothesis that asymptomatic farmers with serum precipitins to farmers' lung disease antigens also have an immune reaction involving the lungs, we did bronchoalveolar lavage (BAL) in 3 groups of dairy farmers. Group 1:7 patients with acute farmers' lung disease. Group 2: 10 asymptomatic farmers with serum precipitins to Micropolyspora faeni. Group 3:9 normal farmers without serum precipitins. Group 1 patients had a large number of cells (90.4 ± 20 × 106) (mean ± SEM) with 72 ± 5.8% lymphocytes in their lavage fluid. Their lavage also had high immunoglobulins A and G/albumin ratios. Six subjects in Group 2 had an increased number of cells (54.1 ± 14.1 × 106) and a high percentage of lymphocytes (52.5 ± 6.6%) in their BAL. Two subjects in Group 3 had similar alterations in their lavage fluid: 60.0 and 69.6 × 106 cells with 20 and 37.5% lymphocytes, respectively. The other subjects in Groups 2 and 3 had normal lavages. Proliferative responses of BAL lymphocytes to phytohemagglutinin was similar i...

Concentration of immunoglobulins in human whole saliva: effect of physiological stimulation.

Abstract: The concentrations of total protein, albumin, IgA, IgG and IgM in human whole salivas were measured with a solid phase radioimmunoassay before and after physiological stimulation of salivary secretion. The geometric mean concentrations (mg/l) before stimulation were: total protein 1600, albumin 60, IgA 140, IgG 16 and IgM 4.1. Physiological stimulation of salivary secretion caused an increase of the total protein concentration to 2400 mg/l, had little effect on the concentrations of albumin and IgG, but lowered the concentrations of IgA and IgM to 56 mg/l and 1.7 mg/l, respectively. These findings indicate that a considerable portion of salivary IgA and IgM are produced locally depending on selective transport and that the release from local storage sites is not increased during stimulation as much as the total volume of the saliva.

Preferential binding of chlordecone to the protein and high density lipoprotein fractions of plasma from humans and other species.

Abstract: The preferential distribution of the relatively nonpolar pesticide chlordecone (CD) to liver rather than to fat tissues in humans suggests that it may be transported in plasma differently from other organochlorine pesticides. The plasma binding of [14C] CD was investigated in vitro in human, rat, and pig plasma and in vivo in rat plasma. Protein and lipoprotein fractions were separated by serial ultracentrifugation. Heparin-manganese precipitation and agarose gel electrophoresis were also carried out to determine whether separation techniques altered CD binding to plasma components. In human plasma, the distribution of [14C] CD among proteins and high density, low density, and very low density lipoproteins (HDL, LDL, and VLDL) was 46, 30, 20, and 6%, respectively. The distribution of cholesterol in the same plasma fractions was 4, 20, 63, and 7%, respectively. In the pig and rat the order of binding was similar to that in humans, with protein greater than or equal to HDL greater than LDL greater than or equal to VLDL. Separation by heparin-Mn precipitation confirmed the results obtained by ultracentrifugation. The distribution of [14C] CD in rat lipoprotein was similar whether the CD was administered in vivo or incubated with plasma in vitro, with approximately 80% bound to HDL, 11% to LDL, and 9% to VLDL in either case. Agarose gel electrophoresis of plasma-bound [14C] CD indicated that albumin was the major component of the protein fraction responsible for CD binding. Preferential binding of CD by albumin and HDL may explain its unusual tissue distribution compared to other organochlorine pesticides such as aldrin and dieldrin, which bind preferentially to VLDL and LDL and distribute preferentially to fat tissues.

Familial Euthyroid Thyroxine Excess: Characterization of Abnormal Intermediate Affinity Thyroxine Binding to Albumin*

Abstract: The abnormal high capacity T4 binding site of familial euthyroid T4 excess was separable from prealbumin and T4-binding globulin but not from albumin. We therefore compared T4 binding by albumin preparations isolated from the sera of normal and affected subjects. By equilibrium dialysis, albumin from affected subjects showed an extra T4 binding site (Kd ∼ 50 nM) in addition to the T4 binding sites of normal albumin (Kd ∼ 4 μM). Comparison of the estimated capacity of the additional site (200 μM) with the molar concentration of albumin suggested that only about one third of albumin molecules from affected subjects contained the extra binding site. Estimates of affinity and capacity were used to derive combining powers for the diverse classes of serum T4 binding sites. From these estimates, it appears that the presence of the abnormal site accounts for the approximate doubling of normal mean total T4 (from ∼ 100 nM or 7.7 μg/dl to ∼ 200 nM or 15.5 μg/dl), in order to maintain a normal free T4 in the face of...