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Showing papers on "Bovine serum albumin published in 1982"


Journal ArticleDOI
TL;DR: It was concluded that this does not necessarily lead to protein preferential hydration and stabilization in NaCl, NaCH3COO, and Na2SO4, since these salts have high surface tension increments.
Abstract: The preferential interactions of proteins with solvent components were studied in concentrated salt by densimetric measurements. Proteins were found to be preferentially hydrated in NaCl, NaCH3COO, and Na2SO4. The resulting unfavorable free-energy change was related to the effects of these salts on solubility and stability of the proteins. This unfavorable free-energy change was correlated with the large, positive surface tension increment of these salts, i.e., their perturbation of surface free energy. On the other hand, KSCN, CaCl2, and MgCl2 showed considerable binding to bovine serum albumin, which could be related to their destabilizing and salting-in effects on macromolecules. Since the last two salts have high surface tension increments, it was concluded that this does not necessarily lead to protein preferential hydration and stabilization.

666 citations


Journal ArticleDOI
TL;DR: The Hp values determined by this method were quite parallel to the results obtained by polyacrylamide gel disc electrophoresis, but remarkably large in cattle suffering from severe inflammatory diseases, such as mastitis, pyometra and traumatic reticulitis.
Abstract: A modification of the method of Tarukoski was employed for the determination of bovine serum haptoglobin (Hp). A mixture of serum sample and diluted methemoglobin (Hb+) solution (30 mg/100 ml) was incubated with o-dianisidine reagent, pH 4.1, at 37°C for 45 min. Peroxidase activity of free Hb was found almost lost in the mixture, in which HpHb complex activity was well maintained. Relatively reliable values were obtained even in hemolyzed serum samples as long as the contaminating Hb was less than 150 mg/100 ml. The Hp values determined by this method were quite parallel to the results obtained by polyacrylamide gel disc electrophoresis. They were small or none in normal cattle, but remarkably large (80-130mg% HbBC) in cattle suffering from severe inflammatory diseases, such as mastitis, pyometra and traumatic reticulitis.

296 citations


Journal ArticleDOI
TL;DR: The features of this system permit pathobiologic investigations of bronchial epithelial cells, e.g., aging, differentiation, and carcinogenesis using conditions that isolate the results from the influence of serum, feeder cells, and other undefined factors.
Abstract: Defined culture conditions for routine clonal growth of normal human adult bronchial epithelial cells have been developed. Serum and feeder cell requirements were abrogated by: (a) optimizing the calcium concentration in nutrient medium, MCDB 151; (b) supplementing with purified factors (epidermal growth factor, 5 ng/ml; insulin, 5 micrograms/ml; transferrin, 10 microgram/ml; hydrocortisone, phosphoethanolamine and ethanolamine, each at 5 x 10(-7) M; and trace elements); and (c) coating the surface of the culture dish with a mixture of fibronectin, collagen, and bovine serum albumin. Endothelial cell growth supplement (100 micrograms/ml) and retinoic acid (3 x 10(-10) M) further enchanced growth, whereas cholera toxin was nonmitogenic and serum supplementation (greater than 2%) markedly reduced the growth rate. Using the defined system, dissociated cultures of bronchial epithelial cells, obtained from more than 15 donors, have been subcultured at clonal densities with a colony forming efficiency of 3 to 4%. In addition, high density cultures have been subcultured more than five times with four to six population doublings per passage. The features of this system permit pathobiologic investigations of bronchial epithelial cells, e.g., aging, differentiation, and carcinogenesis using conditions that isolate the results from the influence of serum, feeder cells, and other undefined factors.

277 citations


Journal ArticleDOI
TL;DR: Differences in the course of alkaline denaturation indicate that only a fraction of strain-specific proteinases is capable to convey long-range effects in the host, suggesting a role of the secretory proteinases in the persistence of yeasts on mucous membranes.
Abstract: Randomly selected strains of Candida albicans were grown with bovine serum albumin (BSA) as a single nitrogen source. From all strains tested, culture supernatant contained carboxyl proteinase (E.C.3.4.23) as has been shown that with hemoglobin as a substrate and by specific inhibition with pepstatin-A. According to the separation pattern of BSA fragments, secretory proteinases from C. albicans belong to at least three groups. We have purified the partially proteolytic enzyme of strain 113 and have compared its properties with those of the totally proteolytic enzyme of strain CBS 2730. Both enzymes have virtually identical molecular weight (ca. 44,000) and cross-react immunologically; they differ in pH optimum, isoelectric point, substrate specificity, and resistance against alkali. IgG1, which is the prevalent immunoglobulin of human serum, was not cleaved by enzyme 113. Immunoglobulins A1, A2 and secretory component were cleaved by both enzymes, which points to a role of the secretory proteinases in the persistence of yeasts on mucous membranes. Differences in the course of alkaline denaturation indicate that only a fraction of strain-specific proteinases is capable to convey long-range effects in the host.

240 citations



Journal ArticleDOI
01 Nov 1982-Cell
TL;DR: Results suggest that thrombospondin and fibronectin may interact during platelet-substratum adhesion or during platelets-platelet aggregation, or both.

166 citations


Journal ArticleDOI
TL;DR: The persistence of leukotriene A4 in the presence of albumin confirms and extends recent observations on its uniform and predictable influence on eicosanoid stability and can no longer be viewed as inert considering its capacity to modify the stability of several, structurally diverse eICosanoids.

152 citations


Journal ArticleDOI
TL;DR: An inhibition immunoassay method developed with the use of microtitre plates coated with a pyridinoline--gelatin conjugate and rabbit antisera directed against pyridsinoline linked to bovine serum albumin is described.
Abstract: An inhibition immunoassay method for the determination of pyridinoline was developed with the use of microtitre plates coated with a pyridinoline--gelatin conjugate and rabbit antisera directed against pyridinoline linked to bovine serum albumin. The sensitivity of the assay is about 2pmol of pyridinoline, and the presence of related pyridinium and lysine-derived compounds does not significantly interfere with the procedure. Its application to tissue and human urine samples is described.

127 citations


Journal ArticleDOI
TL;DR: A hormonally defined medium is described which facilitates the survival of small neurons in primary cultures of mouse cerebellum, showing a tendency to grow in cell clusters with a dense network of processes.

126 citations


Journal ArticleDOI
TL;DR: The collagen matrix culture system allows sustained growth of primary cultures of both normal and neoplastic mammary epithelium in serum-free conditions and will be useful in identifying and investigating the role of hormones, growth factors, and nutritional factors in regulating the growth of mammaries.
Abstract: Freshly isolated normal and tumor mouse mammary epithelial cells embedded within a collagen gel matrix undergo sustained growth when cultured for as long as 3 wk in a serum-free medium composed of a 1:1 (vol/vol) mixture of Hepesbuffered Ham's F12 and Dulbecco's modified Eagle's medium supplemented with insulin, epidermal growth factor (EGF), transferrin, bovine serum albumin fraction V, and cholera toxin. Of these additives, only insulin, EGF, and albumin are required for the growth of most normal cells. Albumin is not always an absolute requirement for growth but greatly enhances it. Lithium has been found to stimulate the growth of normal cells and can replace EGF. The collagen matrix culture system allows sustained growth of primary cultures of both normal and neoplastic mammary epithelium in serum-free conditions. This serum-free system will be useful in identifying and investigating the role of hormones, growth factors, and nutritional factors in regulating the growth of mammary epithelial cells.

125 citations


Journal ArticleDOI
TL;DR: It is possible that the frequent identification of cellular actin in a number of enveloped viruses may be attributed to the interaction of actin and M protein or its equivalent.
Abstract: Evidence for an interaction of the membrane (M) protein of Newcastle disease and Sendai viruses with cellular actin was obtained by three different techniques. M protein linked to Sepharose 4B was found to bind actin, but not myoglobin or bovine serum albumin, and to selectively remove actin from a mixture of these three proteins. Sedimentation of a mixture of M protein and F-actin through a sucrose gradient resulted in sedimentation of M protein with actin. Control proteins, bovine serum albumin and cytochrome c, did not sediment with actin. In circular dichroism studies, M protein added to actin in a 1:1 complex resulted in a significant increase in negative ellipticity at 220 nm, which corresponds to an increase in alpha-helix and a decrease in beta-structure and random coil. This is indicative of an interaction between M protein and actin. It is possible that the frequent identification of cellular actin in a number of enveloped viruses may be attributed to the interaction of actin and M protein or its equivalent.

Journal ArticleDOI
TL;DR: The data indicate that the antibody response to the pure polysaccharides is thymus independent and that this response can be changed into a thymUS-dependent response by covalent conjugation of the poly Saccharide to aThymus-dependent protein.
Abstract: The nature and kinetics of the serum antibody response to pneumococcal type 3 and meningococcal group C polysaccharides and their protein conjugates were studied in mice. Bovine serum albumin and diphtheria and tetanus toxoids were used as carrier proteins. The purified polysaccharides induced only immunoglobulin M (IgM) antibodies in thymus-bearing as well as congenic athymic (nude) mice. The polysaccharides covalently conjugated to proteins produced IgM and IgG antibodies in normal mice, but only IgM antibodies in nude mice. A second dose of the polysaccharide-protein conjugates resulted in a booster effect in the IgG response to the polysaccharides. Moreover, memory B-cells, generated after a primary injection with the polysaccharide-protein conjugates, could be triggered to the production of IgG antibodies after a second injection with the pure polysaccharides alone. These data indicate that the antibody response to the pure polysaccharides is thymus independent and that this response can be changed into a thymus-dependent response by covalent conjugation of the polysaccharide to a thymus-dependent protein.

Journal ArticleDOI
TL;DR: Using the radioimmunoassay it could be shown that endogenous LTC4 is released in a dose‐dependent manner by human polymorphonuclear leucocytes stimulated with the divalent cation ionophore A23187.

Journal ArticleDOI
TL;DR: Results indicate that the LTA molecule is able to bind via its polyanionic backbone to positively charged residues of surface proteins of cells of S. pyogenes.
Abstract: The orientation of lipoteichoic acid (LTA) molecules on the surface of bacterial cells undoubtedly is determined by the ability of the LTA, during its transit through the cell wall, to bind via its polyglycerophosphate backbone or its glycolipid moieties to other constituents of the cytoplasmic membrane and the cell wall. We have investigated the possibility that LTA may become anchored to the cell surface by binding through its polyanionic backbone to positively charged regions of cell wall proteins. LTA was found to prevent the precipitation of partially purified HCl extracts of several strains of streptococci as well as a structurally defined streptococcal M protein molecule (pep M24) in 83% solutions of ethanol. The formation of complexes between LTA and M protein was demonstrated further by immunoelectrophoresis of pep M24 protein with increasing concentrations of radiolabeled LTA and by using antiserum against pep M24 to develop precipitin arcs. Pep M24 electrophoresed alone produced a single precipitin arc close to the origin. In contrast, when electrophoresed as a mixture with LTA or deacylated LTA, the M protein produced a second precipitin arc toward the anode coinciding with the area of migration of the radioactive LTA. Increasing concentrations of LTA or deacylated LTA shifted increasing amounts of the pep M24 antigen to the region of the second arc. Maleylation of M protein to block the positively charged free amino groups before mixing it with LTA prevented the formation of complexes. The complexes formed by the M protein with LTA, but not with deacylated LTA, showed the capacity to bind bovine serum albumin; LTA had been shown previously to bind to the fatty acid binding sites on bovine serum albumin. These results indicate that the LTA molecule is able to bind via its polyanionic backbone to positively charged residues of surface proteins of cells of S. pyogenes. The implications of such interaction as to the orientation of LTA molecules on the surface of cells of S. pyogenes are discussed.

Journal ArticleDOI
TL;DR: Bovine serum albumin (BSA), either untreated or defatted, was found to be better than polyvinylpyrrolidone, ovalbumin, or alpha-lactalbumin, both at stimulating and maintaining motility levels and at reducing the tendency of the washed spermatozoa to stick to glass.
Abstract: Epididymal spermatozoa from rabbit and ram were washed either once or twice using an efficient washing procedure and were then diluted in various media to a final concentration of approximately 1.4 x 10(7) cells/ml and incubated at 30 degrees C for up to 12 hours. Bovine serum albumin (BSA), either untreated or defatted, was found to be better than polyvinylpyrrolidone, ovalbumin, or alpha-lactalbumin, both at stimulating and maintaining motility levels and at reducing the tendency of the washed spermatozoa to stick to glass. BSA was effective in all media tested, being independent of Ca2+, PO4(3-), HCO3-, and ionic strength). BSA has a reversible stimulatory effect on motility. If BSA was added to sperm suspensions 3 1/2 hours after they had been washed and diluted in protein-free medium, motility was stimulated to levels not significantly lower than those observed in samples that had been washed and diluted in the presence of BSA. However, samples washed into BSA and then washed free of it behaved essentially as though they had never been in contact with protein. The motility, survival, and response to BSA of twice-washed spermatozoa were the same as those of once-washed spermatozoa, showing that epididymal plasma factors are not required for survival in vitro. It was concluded that dilution is not essentially detrimental to rabbit and ram spermatozoa. However, severe dilution of semen may result in levels of male reproductive tract fluids insufficient either to stimulate motility or to prevent sticking of motile cells to container surfaces. Few motile spermatozoa are recovered from samples of such diluted semen.

Journal ArticleDOI
01 Feb 1982-Cell
TL;DR: The genome-linked protein (VPg) of poliovirus has been chemically synthesized, coupled to bovine serum albumin carrier and injected into rabbits and an antibody response was elicited not only by the full-length synthetic VPg peptide, but also by a synthetic 14-amino acid carboxy-terminal peptide.

Journal ArticleDOI
TL;DR: It is shown that protein phosphorylation is a necessary step in the process that leads to activation of K+ channels by serotonin in neuron R15 and that this response is mediated by cAMP.
Abstract: Previous work has shown that serotonin induces an increase in membrane K+ conductance in Aplysia neuron R15 and that this response is mediated by cAMP. The present study examines the role of protein phosphorylation in the response to serotonin. A specific inhibitor of cAMP-dependent protein kinase was injected intracellularly into neuron R15. The injection blocked the serotonin-induced increase in K+ conductance completely for at least 4 hours. The blockage was selective because the cell's response to dopamine was not inhibited. Furthermore, the blockage was specifically produced by protein kinase inhibitor because injection of other proteins (alpha-bungarotoxin and bovine serum albumin) did not affect the serotonin response. The serotonin response recovered fully 5-13 hours after the injection, presumably as a result of intracellular proteolysis of the protein kinase inhibitor. The results indicate that protein phosphorylation is a necessary step in the process that leads to activation of K+ channels by serotonin in neuron R15.

Journal ArticleDOI
TL;DR: This stabilized (chiral) LTA4 was converted to LTB4 by an epoxide hydrolase activity in the 100,000 x g supernatant fraction from sonified rat basophilic leukemia cells.

Journal ArticleDOI
TL;DR: No significant stimulatory effect was shown by transferrin, thrombin, L-glutamine, putrescine, selenous acid, dexamethasone, 7S nerve growth factor, or multiplication-stimulating activity.
Abstract: The ability of purified growth factors, insulin, ascorbate, and several other compounds to stimulate DNA synthesis by rabbit articular chondrocytes was studied in monolayer culture. Platelet-derived growth factor (1 U/ml), pituitary fibroblast growth factor (1-100 ng/ml), and epidermal growth factor (1-50 ng/ml) were stimulatory in a basal medium supplemented with 1% heat-inactivated fetal bovine serum. Insulin, 1-50 micrograms/ml, has small growth-promoting effects but acted synergistically with platelet-derived, pituitary fibroblast, and epidermal growth factors. Increasing concentrations of serum up to 10% enhanced the growth-promoting action of the purified factors, but not of insulin. There were indications of cooperation between insulin and bovine serum albumin and dexamethasone. Ascorbate (0.2 mM) reduced or had little growth-promoting action in the basal medium. At 5 and 10% serum concentrations, however, ascorbate promoted DNA synthesis as effectively as the purified growth factors. No significant stimulatory effect was shown by transferrin, thrombin, L-glutamine, putrescine, selenous acid, dexamethasone, 7S nerve growth factor, or multiplication-stimulating activity.

Journal ArticleDOI
TL;DR: Latex immunoassay, a sensitive method based on latex particle agglutination, is used here for the determination of retinol-binding protein in human biological fluids and results are similar to that described previously for beta 2-microglobulin.
Abstract: Latex immunoassay, a sensitive method based on latex particle agglutination, is used here for the determination of retinol-binding protein in human biological fluids. The assay, similar to that described previously for beta 2-microglobulin (Clin. Chem. 27:832-837, 1981), consists of incubating the sample at 37 degrees C for 30 min with antibody-coated particles, then quantifying the resulting agglutination by particle-counting or turbidimetry. Latex particles coated with antibody are stabilized just before use by dispersing them in a solution of bovine serum albumin at pH 10. The standard curve ranges from 0.5 to 32 micrograms/L; recovery averages 102% in urine and 93.5% in serum; between- and within-assay CVs range from 5.1 to 11.7%. The correlation coefficients of latex immunoassay with rocket immunoelectrophoresis for analysis of retinol-binding protein in 26 urines and with radial immunodiffusion in 30 sera are 0.99 and 0.91, respectively. In healthy subjects, the mean urinary excretion of retinol-binding protein is 52.5 micrograms/g of creatinine (SD = 59.2 micrograms/g of creatinine; n = 150) and the concentration in serum averages 46 mg/L (SD = 10.4 mg/L, n = 22).

Journal ArticleDOI
TL;DR: Horseradish peroxidase, assayed with o-phenylenediamine, is irreversibly inactivated when incubated in phosphate buffer, 100 mmol/L, at pH 5. The inactivation was independent of commercial source of the enzyme or the kind of chromogenic assay used.
Abstract: Horseradish peroxidase, assayed with o-phenylenediamine, is irreversibly inactivated when incubated in phosphate buffer, 100 mmol/L, at pH 5. The inactivation depends on both duration and incubation and phosphate concentration. Phosphate was the most potent inactivator and citrate the least potent of a series of buffers tested. The inactivation is not attributable to ionic strength per se or to Na+ or K+. The observed inactivation did not occur at high concentrations (2500 nmol/L, 0.1 g/L) of enzyme; however, this "protective" effect could not be reproduced by adding bovine serum albumin or a surfactant (Tween 20) to lower concentrations of enzyme. The inactivation was independent of commercial source of the enzyme or the kind of chromogenic assay used. On the basis of this information, we optimized the assay so that it gave eightfold greater absorbance values than those reported by others. The improved assay was sensitive to as little as 0.4 pmol/L (16 ng/L) of peroxidase, and was linear over the range of 0.4 to 5 pmol/L (16-200 ng/L).

Journal ArticleDOI
TL;DR: Two serum‐free chemically defined media are developed that support the growth in culture of human diploid fibroblasts to the same extent as Eagle's basal medium supplemented with 10% fetal bovine serum (FBS).
Abstract: We have developed two serum-free chemically defined media (RITC 78-6 and RITC 80-7) that support the growth in culture of human diploid fibroblasts to the same extent as Eagle's basal medium (BME) supplemented with 10% fetal bovine serum (FBS). These two media contain modified Eagle's minimum essential medium (MEM) supplemented with nonessential amino acids, various trace metals, organic compounds and growth factors [insulin, mouse epidermal growth factor (m-EGF), transferrin and triiodothyronine (T3)]. RITC 80-7 medium differs from RITC 78-6 in that it contains thymidine, hypoxanthine, and vitamin B12 and supports the long-term serial cultivation of human diploid cultures. The addition of commercial bovine serum albumin (BSA, 5 g/liter) to the medium enhances cell growth. This effect is not observed if BSA is first delipidized, but reconstitution of BSA with certain lipids restores its ability to promote growth. BSA has an inhibitory effect on cellular attachment but this is overcome when fibronectin (FN, 10 mg/liter is added to the medium.

Journal ArticleDOI
TL;DR: In this article, Ross et al. showed that the radius of the effective hard spherical particle representing BSA varies with the net electrical charge of the BSA molecule in a manner which may be calculated from electrostatic theory.
Abstract: The light scattering of bovine serum albumin (BSA) has been measured at protein concentration up to 90 g/L and at pH values between 4.4 and 7.6. The dependence of scattering on both protein concentration and pH may be quantitatively accounted for by a simple extension of the hard-sphere model for protein solutions [Ross, P. D. & Minton, A. P. (1977) J. Mol. Biol.112, 437–452] allowing for electrostatic repulsions between molecules. According to the extended model, the radius of the effective hard spherical particle representing BSA varies with the net electrical charge of the BSA molecule in a manner which may be calculated from electrostatic theory.

Journal Article
TL;DR: Human T cell growth factor has been purified more than 800-fold from serum-free lymphocyte conditioned media by utilizing ion exchange chromatography with DEAE-Sepharose, gel filtration with Ultrogel AcA54, and preparative SDS-polyacrylamide gel electrophoresis.
Abstract: Human T cell growth factor (TCGF) has been purified more than 800-fold from serum-free lymphocyte conditioned media by utilizing ion exchange chromatography with DEAE-Sepharose, gel filtration with Ultrogel AcA54, and preparative SDS-polyacrylamide gel electrophoresis. This mitogenic protein, which is released by T cells into the incubating media after exposure to a lectin (PHA), has a m.w. of 13,000 as determined by SDS-PAGE and 20,000 to 25,000 on gel filtration and has an isoelectric point of 6.8. The material extracted from acrylamide gels is a single band (or two nearly superimposed bands) when rerun on analytical SDS gels. Partially purified material is unstable even at -70 degrees C and requires the addition of bovine serum albumin or polyethylene glycol to maintain biologic activity. It is sensitive to proteolytic digestion but resistant to nucleases and thiol-reducing agents such as dithiothreitol. Human TCGF can be reversibly denatured with urea or SDS. Material extracted from acrylamide gels has been shown to sustain T lymphoblasts in tissue culture. In contrast to lectins, certain antigens, and crude lymphocyte conditioned media, purified TCGF does not initiate lymphocyte blastogenesis but is a highly selective mitogen for T cells previously activated by exposure to lectins or antigens.

01 Jan 1982
TL;DR: The observed inactivation of horseradish peroxidase did not occur at high concentrations and could not be reproduced by adding bovine serum albumin or a surfactant to lower concentrations of enzyme, independent of commercial source of the enzyme or the kind of chromogenic assay used.
Abstract: Horseradish peroxidase, assayed with o-phenylenediamine, is irreversibly inactivated when incubated in phosphate buffer, 100 mmol/L, at pH 5. The inactivation depends on both duration of incubation and phosphate concentration. Phosphate was the most potent inactivator and citrate the least potent of a series of buffers tested. The inactivation is not attributable to ionic strength per se or to Na+ or K+. The observed inactivation did not occur at high concentrations (2500 nmol/L, 0.1 g/L) of enzyme; however, this “protective” effect could not be reproduced by adding bovine serum albumin or a surfactant (Tween 20) to lower concentrations of enzyme. The inactivation was independent of commercial source of the enzyme or the kind of chromogenic assay used. On the basis of this information, we optimized the assay so that it gave eightfold greater absorbance values than those reported by others. The improved assay was sensitive to as little as 0.4 pmol/L (16 ng/L) of peroxidase, and was linear over the range of 0.4 to 5 pmol/L (16-200 ng/L).

Journal ArticleDOI
TL;DR: The results emphasize the similarity of the structural requirements for the inhibition of matrix synthesis and the stimulation of resorption by active vitamin D metabolites in bone and support the importance of the 1-hydroxyl function to the biologic activity of vitamin D in the skeleton.
Abstract: The effects of selected vitamin D3 metabolites and analogs on bone collagen synthesis in vitro were examined in organ cultures of neonatal mouse calvarial bone. The incorporation of [3H]proline into the collagenase-digestible fraction of newly synthesized protein was progressively inhibited by 1α,25-dihydroxyvitamin D3 (1α,25-(OH)2D3) (10−12 M to 10−7 M) in 24-h cultures, and incorporation into noncollagen protein was also blunted at the higher doses employed. The synthetic analog 1α-hydroxyvitamin D3 (1α-OHD3) was almost 300-fold less potent an inhibitor of collagen synthesis than was 1α,25(OH)2D3, and the natural metabolites 25-hydroxyvitamin D3 (25OHD3) and 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3), 1000-fold less potent, although the dose-response curve for each of these compounds was not parallel with that for 1α,25(OH)2D3. The 24S,25(OH)2D3 enantiomer was four-fold less potent than 24R,25-(OH)2D3 or 25OHD3, and vitamin D3 showed less than 2% the activity of 25OHD3. The responses were unaffected by the substitution of 0.4% bovine albumin for 5% horse serum in the medium, and no stimulation of collagen synthesis was observed in response to 25-hydroxylated metabolites between 2×10−14 and 2×10−6 M or in cultures treated for up to 96 h with 24R,25(OH)2D3 (2×10−10M). The overall results emphasize the similarity of the structural requirements for the inhibition of matrix synthesis and the stimulation of resorption by active vitamin D metabolites in bone. In addition, these studies support the importance of the 1-hydroxyl function to the biologic activity of vitamin D in the skeleton.

Journal Article
TL;DR: 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.
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.

Journal ArticleDOI
TL;DR: A direct radioimmunoassay for serotonin (5-hydroxytryptamine) in cerebrospinal fluid, platelet-poor plasma, and serum is described and polyethylene glycol, alone or in combination with anti-rabbit immunoglobulins, is used to separate bound and unbound tritiated serotonin.
Abstract: We describe a direct radioimmunoassay for serotonin (5-hydroxytryptamine) in cerebrospinal fluid, platelet-poor plasma, and serum. We raised antisera in rabbits against serotonin diazotized to a conjugate of bovine albumin and D,L-p-aminophenylalanine. Polyethylene glycol, alone or in combination with anti-rabbit immunoglobulins, is used to separate bound and unbound tritiated serotonin. The minimum concentration of serotonin detectable is 2 nmol/L in a 200-microL sample. Within-day precision (CV) is 4.3%, between-day precision 7.7%. Analytical recoveries of serotonin are 109% and 101% for cerebrospinal fluid and plasma, respectively. Tryptophan, 5-hydroxytryptophan, 5-hydroxyindoleacetic acid, and 5-hydroxytryptophol do not interfere with the assay. However, 5-methoxytryptamine and tryptamine cross react. Of samples of cerebrospinal fluid from patients with disc herniations (n = 21) or low-pressure hydrocephalus (n = 10), one-third had concentrations of 2-4 nmol/L and two-thirds were below the minimum detectable concentration. The observed range for the concentration of serotonin in plasma of 14 normal subjects was 5-14 nmol/L (mean +/- SD, 9 +/- 3 nmol/L). The observed ranges for serotonin in serum were: for 10 women 520-900 (mean +/- SD: 695 +/- 110) nmol/L and for 10 men 380-680 (520 +/- 94) nmol/L.

Journal ArticleDOI
TL;DR: It is concluded that charge is an important determinant of protein entry into the CSF.
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.

Journal Article
TL;DR: Bacterial culture filtrates and peptides, which were potent chemotaxins for leukocytes from other species, failed to affect bovine leukotaxis, indicating that bovines have chemotactic requirements distinct from those of other species.
Abstract: A system was described for studying bovine leukotaxis. Chemotaxis was readily observed toward bovine serum activated by zymosan of agarose. However, bacterial culture filtrates and peptides, which were potent chemotaxins for leukocytes from other species, failed to affect bovine leukotaxis. Using conditions suitable for studying binding to leukocytes from other species, specific binding of a radiolabeled chemotactic peptide to bovine leukocytes was not apparent. These data indicate that bovine leukocytes have chemotactic requirements distinct from those of other species.