The Metabolism of Serum Proteins in Neonatal Rabbits

1. Incorporation of S³⁵-labeled amino acids into serum proteins has been studied in neonatal and developing rabbits. It was found that, per unit weight, neonatal rabbits synthesized only about 1/36 of the gamma globulin, 1/7 of the beta globulin, ½ of the alpha globulin, and ⅛ of the albumin that an adult synthesized. The growing rabbit developed the ability to synthesize various serum proteins at different times. 2. Plasma volumes and serum protein concentrations were determined at different times during the growth period of the rabbit. Plasma volumes were found to be 1 and ½ times larger in newborn animals than in adults, with a gradual decline to the adult level. The total serum protein concentration at birth was about 60 to 65 per cent of the adult value and gradually increased with growth as the plasma volume decreased. 3. Half-lives of homologous albumin and gamma globulin were studied. The half-life of albumin in neonates was nearly twice as long as the half-life in adults, the latter value being reached at 1 month of age. The half-life of gamma globulin in neonates was more than twice as long as the half-life in adults and reached adult values at 2 to 3 months. 4. Attempts were made to alter serum protein metabolism. Gamma globulin synthesis early in life was augmented with antigen injections.     

Variation in blood serum proteins and association with somatic cell count in dairy cattle from multi-breed herds

Blood serum proteins are significant indicators of animal health. Nevertheless, several factors should be considered to appropriately interpret their concentrations in blood. Therefore, the objectives of this study were (1) to assess the effect of herd productivity, breed, age and stage of lactation on serum proteins and (2) to investigate association between serum proteins and somatic cell count (SCC) in dairy cattle. Milk and blood samples were collected from 1508 cows of six different breeds (Holstein Friesian, Brown Swiss, Jersey, Simmental, Rendena and Alpine Grey) that were housed in 41 multi-breed herds. Milk samples were analyzed for composition and SCC, while blood samples were analyzed for serum proteins (i.e. total protein, albumin, globulin and albumin-to-globulin ratio (A : G)). Herds were classified as low or high production, according to the cow’s average daily milk energy yield adjusted for breed, days in milk (DIM) and parity. Data were analyzed using a linear mixed model that included the fixed effects of DIM, parity, SCS, breed, herd productivity and the random effect of the Herd-test date within productivity level. Cows in high producing herds (characterized also by greater use of concentrates in the diet) had greater serum albumin concentrations. Breed differences were reported for all traits, highlighting a possible genetic mechanism. The specialized breed Jersey and the two dual-purpose local breeds (Alpine Grey and Rendena) had the lowest globulin concentration and greatest A : G. Changes in serum proteins were observed through lactation. Total protein reached the highest concentration during the 4th month of lactation. Blood albumin increased with DIM following a quadratic pattern, while globulin decreased linearly. As a consequence, A : G increased linearly during lactation. Older cows had greater total protein and globulin concentrations, while albumin concentration seemed to be not particularly affected by age. A linear relationship between serum proteins and SCS was observed. High milk SCS was associated with greater total protein and globulin concentrations in blood. The rise in globulin concentration, together with a decrease in albumin concentrations, resulted in a decline in A : G as SCS of milk increased. In conclusion, such non-genetic factors must be considered to appropriately interpret serum proteins as potential animal welfare indicator and their evaluation represents an important first-step for future analysis based on the integration of metabolomics, genetic and genomic information for improving the robustness of dairy cows.     

ALTERATIONS IN SERUM PROTEIN LEVELS IN ACUTE HUMAN POLIOMYELITIS: RATIONALE FOR THERAPY

Serial determinations of serum protein levels in acute human poliomyelitis revealed a progressive drop of the serum albumin level which bore close relationship to the amount of clinical paralysis. This loss of serum albumin began about the third day after onset of clinical symptoms and progressed to the tenth day or longer. The more severe the clinical involvement, the less was the tendency to spontaneous correction of the albumin deficiency. Declining serum albumin levels were concomitant with progressively rising serum globulin values.When pooled irradiated human blood plasma was administered, the depleted serum albumin levels were stabilized or made to approach normal, depending upon the severity of clinical involvement. It is felt that the administration of blood plasma resulted in definite clinical benefit with regard to the severity, extent, and duration of paralysis.     

Metabolic interactions between animal cells through permeable intercellular junctions.

The isolated perfused rat liver was used to study the degradation of ¹²⁵I-labelled protein supplied in the perfusion medium. Formaldehyde-denatured proteins (human serum albumin, bovine serum albumin and especially rat liver phosphoenolpyruvate carboxykinase (GTP)) were taken up by the liver and degraded at high rates. Native human serum albumin was not degraded at significant rates by the perfused liver, while native phosphoenolpyruvate carboxykinase (GTP) was catabolised at about one-fourth the rate of the denatured enzyme. The degradation rate of denatured human serum albumin increased markedly as protein was added up to 0.7 mg, and more gradually with further increases in added protein. The biphasic nature of concentration dependence probably reflects the contribution of different cell types in the liver. Autoradiographic examination of serial biopsies taken during perfusion of the liver with formaldehyde-denatured, ¹²⁵I-labelled bovine serum albumin showed that at the cellular level the radioactivity was located predominantly in Kupffer and other non-parenchymal cells; and at the subcellular level the radioactivity was largely in endocytic vesicles, lysosomes and occasionally in the sinusoidal spaces. No significant radioactivity was found associated with other cytoplasmic organelles or the nucleus. It is concluded that lysosomes of the non-parenchymal cells are primarily responsible for the degradation of denatured extracellular protein that enters the liver.     

Serum Protein Electrophoresis in Tuberculosis

Serial serum protein electrophoretic determinations at two-month intervals were carried out on 84 tuberculous patients undergoing treatment in a sanatorium. An attempt was made to correlate changes in the albumin/alpha-2 globulin ratios and gamma globulin levels with clinical and roentgenographic status. It was observed that before treatment of the tuberculous process the albumin/alpha-2 globulin ratios were low and the gamma globulin levels were increased. As improvement occurred, albumin/alpha-2 globulin ratios increased and gamma globulin values decreased. A poor prognosis was indicated by decreasing albumin/alpha-2 globulin ratios. Electrophoretograms are of particular value in assessing the effect of surgery, especially when roentgenographic studies are not informative. They are also a valuable guide in deciding upon the duration of therapy necessary for individual cases.     

The separation of intracellular serum albumin from rat liver

1. Antibody precipitation of serum albumin from rat liver extracts yields impure preparations of the protein. 2. When rat liver is labelled with l-[1-(14)C]leucine, antibody precipitation of albumin leads to material that is contaminated with a protein or proteins of very high specific radioactivity. Only 10-25% of the radioactivity of the antibody precipitate is associated with serum albumin. 3. A chromatographic procedure is described that can be used to separate radiochemically pure serum albumin from antibody precipitates obtained from extracts of rat liver. 4. Extracellular albumin secreted by liver slices yields a precipitate with antibody which contains much less radioactive impurity. About 70-90% of the radioactivity is associated with serum albumin. Serum albumin separated by antibody precipitation from rat serum labelled in vivo was not contaminated with the radiochemical impurities associated with intracellular albumin. 5. A simple method is described of obtaining the content of serum albumin in rat liver extracts by the technique of isotope dilution and ion-exchange chromatography.     

Atlantic salmon (Salmo salar) serum albumin: cDNA sequence, evolution, and tissue expression

Atlantic salmon serum albumin is one of the most abundant proteins in salmon liver, representing 1.6% of all clones in a cDNA library made from salmon liver RNA. The DNA from a number of clones was sequenced to reveal an open reading frame of 1,827 bases encoding a 608-amino-acid protein. The sequenced 5' untranslated region is 69 bases long and the 3' untranslated region contains two putative polyadenylation signals and poly(A) tail. Sequence analysis of different clones indicates the presence of a second cDNA for salmon serum albumin. Multiple alignments of salmon serum albumin deduced amino acid sequence with Xenopus laevis, rat, bovine, and human serum albumins shows significant conservation of cysteine residues. The triple domain structure of serum albumin proteins is maintained. Unlike mammalian systems where serum albumin expression appears to be specific to liver only, salmon serum albumin is expressed in muscle also.     

Attempted Passive Immunization of Young Calves Against Eimeria bovis

SYNOPSIS. Two experiments attempted to produce passive immunity against Eimeria bovis coccidiosis in Holstein-Friesian calves. Immune serum concentrated by a freezing technique, or serum globulin obtained by a precipitating technique from immune calves, was injected intravenously or intraperitoneally into young calves. Four calves received concentrated immune serum injected intravenously on the day of oral inoculation with sporulated oocysts and again 7 and 14 days later. Four calves were given intravenous injections with some of the same serum on the 7th and 14th days after inoculation and 4 others were given a single similar injection with the same serum 14 days after inoculation.
Three calves in a second experiment received intraperitoneal injections of serum globulins in increasing amounts every 3 days for 2 weeks. The calves were then orally inoculated with sporulated oocysts one week after the last globulin injection. Some calves receiving immune serum had an anaphylactoid reaction characterized by increased respiration rate, dyspnea, coughing, and salivation; however, all affected calves recovered spontaneously within 2 hours. Calves receiving serum globulin had no reactions.
Coccidiosis developed in all of the calves in spite of the injection of immune serum or globulin presumed to carry the immune factor. There was no detectable difference in the rate of oocyst discharge or in clinical symptoms between treated and control calves; therefore, no evidence of passive immunity was observed.     

Serum proteins in young hedgehogs

Four serum protein components are present at birth: α-, β- and γ-globulins and albumin.
The preferential transfer of γ rather than β-globulin, which occurs both prenatally and postnatally in this species, is reflected in the initial amounts of these proteins present at birth and in the relatively rapid increase in the concentration of γ-globulin which occurs postnatally. By five weeks of age seven protein components are discernible in the serum, which is comparable in this respect to adult serum.
The albumin/globulin ratio changes from about 1.5 at birth to about 0.3-0.4 in the active adult.     

THE EFFECT OF PURE PROTEIN SOLUTIONS AND OF BLOOD SERUM ON THE DIFFUSIBILITY OF CALCIUM

1. A comparative study has been made of the diffusibility of calcium in solutions of crystalline egg albumin, serum globulin, and human blood serum. 2. In all three of these solutions, at pH 7.4, molal Ca concentrations within the membrane are greater than the calcium concentrations in the outside solutions, quite in accordance with the Donnan theory. 3. At pH 7.4, the ratio of See PDF for Structure varies directly with the protein concentration whether the solution be one of egg albumin, serum globulin, or blood serum. This is also in accordance with the Donnan theory. 4. On the acid side of the isoelectric point of the proteins, the concentration of Ca outside becomes greater than the concentration in the solution of blood serum or pure protein, as is demanded by the Donnan theory. 5. The magnitude of the Ca ratios on the alkaline and acid sides of the isoelectric points is probably the resultant of the Donnan equilibrium and the formation of complex Ca-protein ions. Northrop and Kunitz have shown the probability of the existence of such ions in the case of Zn⁺⁺, K⁺, and Li⁺, where satisfactory electrodes have been developed for E.M.F. measurements.     

Influence of hydrocortisone acetate administered to the lactating rat on protein and lactose content in milk and serum protein, glucose and insulin levels in dams and pups

The influence of i.m. administration to the mother of hydrocortisone acetate (doses of 0.4, 0.8 or 2.0 mg/100 g body weight/day) during the first 15 days of lactation on milk protein and lactose composition and serum levels of protein, glucose and insulin in dams and pups is studied. Total serum proteins and albumin/globulin ratio in dams were unchanged by treatment. The daily injection of 0.4 or 0.8 mg/100 g body weight failed to alter serum levels of glucose or insulin in dams, whereas a dose of 2.0 mg/100 g body weight led to a rise in glucemia (from 118 +/- 3.2 to 133 +/- 5.3) which was accompanied by a sharp change in insulinemia (from 40.7 +/- 4.1 to 83.6 +/- 6.9). All three doses raised protein levels in milk. The smallest increase was recorded with 2.0 mg/100 g body weight; this dose also reduced milk lactose content. Total serum proteins in pups rose slightly but nonsignificantly, and no significant effects were noted on albumin/globulin ratio or serum glucose and insulin levels.     

Albumin and transferrin synthesis during development in the rat

In this study, the incorporation of [(14)C]leucine into albumin and transferrin in early rat foetuses, vitelline plus amniotic membranes, chorioallantoic placenta and perinatal rat liver slices was measured and used to detect and compare the rates of synthesis of the two proteins. Albumin synthesis was detected in the body of foetuses from 13 days gestation onwards. Transferrin synthesis was detected only after day 15. Transferrin synthesis was demonstrable in the membranes but not in the chorioallantoic placenta of all the animals investigated, i.e. from 13 to 19 days gestation. Synthesis of albumin and transferrin by the liver of near-term and postnatal animals was shown to correlate with published data on the parenchymal cell number/unit wet wt. of liver. Near-term foetuses synthesized relatively more transferrin than albumin when compared with 10-day postnatal animals. The serum concentrations of the two plasma proteins were also determined. These increased before term whereas the rate of synthesis of albumin and transferrin declined. Postnatally, plasma albumin concentration increased but transferrin concentration decreased, yet the rates of synthesis of both proteins by the liver increased with age. This lack of correlation between the rates of synthesis of the two proteins and their respective plasma concentrations could be explained in part by their increased stability after birth. There was also evidence that the liver haemopoietic cells took up transferrin although they do not synthesize the protein. Thus the decrease in this population of cells during development could also contribute to the discrepancy between liver synthesis and serum concentrations of transferrin.     

The Lack of Selective Reincorporation into Tissue Protein of the Amino Acids Derived from the Catabolism of Serum Protein

Homologous S³⁵-labeled albumin, gamma globulin, and alpha-beta globulin were transfused into rabbits and the specific activities of the electrophoretic fractions of the sera of the recipients were determined at various time intervals up to 12 days after injection. Detectable reincorporation into a fraction other than that transfused was found only in the gamma globulin fraction after albumin injection. This activity rose between 2 and 12 days and reached a level of 2 to 3 per cent of the extrapolated zero time activity of the albumin fraction. When homologous serum protein doubly labeled with I¹³¹ and S³⁵ was transfused into mice, marked drops in the ratios of I¹³¹ to S³⁵ in the serum and tissue proteins were observed between 1 and 48 hours after injection. On the basis of a determination of the absolute and relative amounts of I¹³¹ and S³⁵ found in the various tissue and serum proteins, the amount of reincorporation of S³⁵ into each protein was calculated. The relative amounts of reincorporation of S³⁵ among the various tissues were remarkably similar to the relative amounts of incorporation of S³⁵ after the injection of labeled free amino acids. It is concluded that serum protein does not form a major direct source of amino acids to the tissues but feeds them indirectly through the extracellular pool.     

Rat sulfite oxidase antibodies cross-react with two gene family-related proteins: albumin and vitamin D-binding protein.

Screening lambda cDNA libraries from rat liver with antibody to native rat liver sulfite oxidase (RLSO) showed cross-reaction with two proteins that belong to the same gene family: serum albumin and vitamin D-binding protein. Antibodies raised against native RLSO or sodium dodecyl sulfate-denatured protein cross-reacted with these proteins by Western blot analysis. The relative effectiveness of RLSO antibody binding was estimated to be 1/5 for rat serum albumin and 1/10 for rat vitamin D-binding protein. This result was not caused by contaminating proteins in the RLSO used for immunization as the RLSO preparation did not react with rat serum albumin antibody. RLSO antibodies, selected for their ability to bind rat serum albumin immobilized on nitrocellulose, recognized both rat serum albumin and RLSO. RLSO antibody, with albumin-reactive antibody removed, still recognized vitamin D-binding protein, suggesting that multiple determinants specific to each protein are involved in the cross-reaction. Comparison of RLSO antibody binding to the rat and human proteins indicated that the determinants were species-specific. cDNA clones identified by screening cDNA libraries with RLSO antibody demonstrated that these determinants reside in the C-terminal domain of these proteins. These results suggest that these proteins contain some common immunological features and may be evolutionarily related.     

Changes in serum and plasma proteins and in IgG and IgM antibodies in calves experimentally infected with sarcocystis from dogs.

Fifteen calves were used in three experiments to determine changes in serum and plasma proteins and IgG and IgM levels after oral inoculation with Sarcocystis sporocysts from dogs. Total serum or plasma protein levels in inoculated calves decreased during the acute phase of infection (4 to 5 weeks after inoculation) and then increased and became greater than the levels of control calves 7 to 8 weeks after inoculation. The initial decrease in total protein reflected reduced serum albumin and the subsequent increase reflected increased immunoglobulin levels. Immunoglobulin levels increased in both IgM and IgG fractions. Specific antibody activity against Sarcocystis antigen was 6 to 9 times greater in the IgM than in the IgC fraction 5 weeks after inoculation, but IgG activity became approximately 17 to 27 times greater than IgM activity 10 to 13 weeks after inoculation.     

A study of the uterine protein variations during the estrus cycle in the cow: a comparison with the serum proteins

To investigate uterine protein changes during the estrus cycle in the bovine, 115 pluriparous genital tracts and blood samples were collected from the abattoir in Urmia. Genital tracts were considered healthy based on gross examination of the uterus and uterine histopathological findings. The phase of the estrus cycle was determined by the examination of the structures present on the ovaries and the uterine tonicity. Of the collected samples, 24 were pro-estrus, 21 estrus, 24 met-estrus and 46 diestrus. The uterus was incised and uterine fluid was collected by gentle scraping of the uterine mucosa with a curette. The total protein concentration, protein profiles (on agarose gel electrophoresis) in the uterine fluid were evaluated and compared with those of the serum. Total protein, alpha1, alpha2, beta1 and beta2 globulin values in the uterus were significantly higher than those of the serum (P<0.05), while, the albumin, gamma1 and gamma2 globulin values in the serum were higher than those of the uterus throughout the cycle. During pro-estrus, uterine fluid beta2 (1.96 g/dl) and serum gamma1 (1.07 g/dl) and gamma2 (1.27 g/dl) globulins were higher than those in the other phases of the cycle. During estrus, serum total protein was lower than the other phases (4.92 g/dl), which was considered to be due to a reduction in serum alpha1 (0.25 g/dl), gamma1 (0.65 g/dl) and gamma2 (0.64 g/dl) globulins in this phase. In met-estrus uterine fluid beta1 globulin was in the lowest (1.19 g/dl) and serum gamma2 globulin at a high level (1.24 g/dl). It was concluded that uterine proteins as well as serum proteins fluctuate during the estrus cycle and, except for the albumin and gamma globulins, its protein content is higher than the serum. During the follicular phase of the cycle uterine alpha globulins are higher than those in other phases, with an elevation in beta1 and a reduction in beta2 and gamma globulin values during estrus, which may reflect the preparation of the uterus for receiving spermatozoa for the conception in this phase.     

Maintenance of plasma-derived proteins at much lower concentrations in the uterine lumen of the rabbit: a kinetic study of passage.

Human serum albumin (HSA) and human gamma globulin (HGG) in serum and uterine fluid of nonpregnant rabbits at various times after an i.v. injection (100 mg/kg) were measured by a radial immunodiffusion test using specific antisera. The HSA concentration in uterine fluid rose to a peak at 12 hr when it was 11% of the serum concentration and then declined, whereas HGG reached a peak at 18 hr (3.2% of serum level) and decreased thereafter. The HSA passed 2 1/2 times faster than HGG, but both proteins equilibrated with uterine fluid in about 12-18 hr. Steady state levels of HSA and HGG indicated that uterine fluid: serum ratios were 1:10 and 1:20, respectively. Similar ratios were found for total protein and rabbit serum albumin (1:10) and rabbit gamma globulin (1:20). Therefore, except when there is a local immune response, the uterine lumen contains only about 5% of the serum antibody concentration. Available data in the mouse, rat and dog also indicate disparity between serum and uterine fluid protein levels.     

The effect of soluble rat liver proteins on the activity of microsomal stearoyl-CoA and linoleoyl-CoA desaturase.

1. The influence of bovine serum albumin and soluble rat liver proteins on the activity of rat liver microsomal delta9 and delta6 desaturases has been studied. 2. In the absence of bovine serum albumin, the delta9 desaturase which converts stearoyl-CoA into oleoyl-CoA, shows a non-linear correlation between enzyme activity and protein concentration. 3. Optimum concentrations of bovine serum albumin have three main effects on the enzyme activity: (i) establishes a linear relationship between enzyme activity and protein concentration, (ii) stimulates the enzyme activity 2--3-fold and (iii) raises the optimum substrate concentration from 10 to 100 muM. 4. A highly purified soluble liver protein of molecular weight 24 000 also stimulated the enzyme activity and brought about a linear relationship between enzyme activity and protein concentration. 5. It was concluded that the non-linear kinetics were due to limiting amounts of substrate binding protein in the microsomal preparations. 6. The delta6 desaturase which converts linoleoyl-CoA into gamma-linolenoyl-CoA was also stimulated by bovine serum albumin and soluble liver proteins. 7. The significance of the fatty acid-binding proteins is discussed.     

The Fibrinogen Concentration in Blood of Dairy Cows and its Influence on the Interpretation of the Glutaraldehyde and Formol-Gel Test Reactions

It has earlier been shown that the formol-gel test on serum and glutaraldehyde test on whole blood are simple and rapid methods for evaluation or the immunoglobulin status in the cow. Both tests function as coagulation tests in which aldehyde groups oross-link basic blood globulins at their NH2-groups, forming polymerisates. The glutaraldehyde has in whole blood the capacity to polymerize not only immunoglobulins but also fibrinogen. This investigation was made in order to study whether the fibrinogen level may influence the result of the glutaraldehyde test, so revealing any differences between the results of that and the formol-gel test carried out on serum. In 92 cows with a variety of clinical disorders (most of them with inflammatory processes) the total protein, albumin, total globulin concentration and albumin/globulin ratio in serum and fibrinogen concentration in plasma were recorded. The material was grouped according to glutaraldehyde and formol-gel test reactions. It is shown that increases in the fibrinogen level have an effect on the results of the glutaraldehyde test. A positive glutaraldehyde test in more acute processes is ascribed to a heavy rise of plasma fibrinogen in its capacity of acute-phase protein. A positive glutaraldehyde test in chronic diseases may be viewed as a result of interaction between high immunoglobulin concentrations and elevated fibrinogen concentration. In conclusion the fibrinogen and immunoglobulin status of blood is important to assess in many diseases of cattle. The semiquantitative tests described for field use can separately, or especially in parallel use, provide valuable information about the character and development of a disease and may be regarded as good substitutes for the sedimentation rate (SR), which is not demonstrable in cattle. kw|Keywords|k]bovine fibrinogen; k]bovine serum proteins; k]formol-gel reaction; k]glutaraldehyde test; k]acute and chronic inflammations     

Carbohydrate, protein and glycoprotein metabolism by the guinea-pig liver in chronic metribuzin poisoning

Male guinea-pigs weighing 400-600 g, 8 months old, were given metribuzin directly into the gastric lumen over a period of 30 days (20 animals) or 90 days (20 animals) 6 times a week. In the liver of the poisoned animals, the glycogen level and the AspAT and AlAT activities, while in the serum the total protein and the fractions albumin, alpha 1-globulin and gamma-globulin significantly decreased; serum glucose and the serum fractions alpha 2-globulin and beta-globulin, each showed an increase. The glycogen level in the liver, total protein, glucose as well as the alpha 1 and alpha 2 globulin fractions in the serum showed not appreciable difference between 30 and 90 days of intoxication. After 90 days of metribuzin treatment AspAT and AlAT dropped in the liver and rose in the serum, in comparison to the 30-day values. As to the parameters of glycoprotein metabolism, the intoxicated animals showed a significant decrease and increase in concentration of hexosamines and sialic acids in the liver and serum, respectively. Metribuzin intoxication also cause a significant decrease in activity of glucosamine phosphate isomerase and significant increase in activity of glycosidases in the liver. The results suggest that metribuzin disturbs the metabolism of carbohydrates, proteins and glycoproteins in the guinea-pig liver.