Serum proteins in the leopard seal, Hydrurga leptonyx, in Prydz Bay, Eastern Antarctica and the coast of NSW, Australia

Blood protein analysis including total serum protein and albumin by chemical methods, fibrinogen estimation and serum protein electrophoresis (SPE) was performed on the leopard seal, Hydrurga leptonyx. The most commonly observed SPE pattern was eight fractions designated albumin, alpha(1a), alpha(1b), alpha(2a), alpha(2b), beta(1), beta(2) and gamma-globulin. Significantly higher total serum protein and albumin concentrations, as determined by chemical methods, and significantly higher alpha(2)-globulin concentrations, determined by SPE, were seen in free-ranging male seals compared to females, whilst significantly higher beta-globulin concentrations were seen in female seals. Season of sampling influenced fibrinogen and beta(2)-globulin concentrations, whereas there were no significant differences in any protein concentrations with moult status. Qualitative comparison of SPE traces of leopard seals in Antarctica with "sick" individuals in NSW, Australia revealed obvious differences, as did quantitative comparison of protein concentrations where differences in alpha(1), alpha(2), beta(1), beta(2), and gamma-globulin concentrations were seen. These findings suggest that SPE is a useful tool for investigating serum proteins in the leopard seal, with applications for the investigation of "sick" individuals and the assessment of variation in homeostasis. This technique could also be used to identify the presence of environmental stressors, subclinical disease and physiological variation within specific seal populations.     

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.     

Seasonal changes in blood serum protein fractions and in activity of AspAT and AlAT in Arabian brood mares and their foals

In 34 pure breed Arabian horses divided into four groups (Gr. I--10 pregnant mares, Gr. II--7 barren mares, Gr. III--10 foals born in 1981, Gr. IV--7 foals born in 1982) seasonal changes in total blood serum protein, its electrophoretic fractions and the activity of AspAT and AlAT were studied. Seasonal cyclicity was found in all groups in the amount of total serum proteins, and alpha 2- and beta 1-globulin fractions. Cyclicity was found in the level of albumin and activity of AspAT in three groups, not Gr. II, and in gamma-globulin, not Gr. IV. beta 2-globulin and AlAT cyclicity was found in two groups and alpha 1-globulin cyclicity was found only in Gr. II. Out of nine indices studied, cyclicity was found in eight of them in pregnant mares, Gr. I; in seven in older foals, Gr. III; in six in the young foals, Gr. IV; and in five in barren mares, Gr. II.     

Hematological, protein electrophoresis and cholinesterase values of free-living nestling peregrine falcons in Spain

Protein electrophoresis, hematological and cholinesterase values were determined in 32 nestling free-living peregrine falcons (Falco peregrinus) (15- to 27-days-old) in order to establish normal reference values for this population. The following values (mean +/- SD) were observed: prealbumin 0.31 +/- 0.04 g/dl, albumin 1.25 +/- 0.06 g/dl, alpha1 and alpha2-globulin 0.23 +/- 0.02 and 0.16 +/- 0.02 g/dl respectively, beta-globulin 1.02 +/- 0.05 g/dl, gamma-globulin 0.060 +/- 0.08 g/dl, total protein 3.79 +/- 0.18 g/dl, 21.26 +/- 1.30 white blood cells/microl (1 x 10(3)), 2.17 +/- 0.07 red blood cells/microl (1 x 10(6)), packed cell volume 37.58 +/- 0.82%, hemoglobin 20.96 +/- 0.29 g/dl, heterophils 61.14 +/- 2.50% and cholinesterase 1,184 +/- 75 IU/L. There were no difference in any of these parameters among males and females. The hematological values obtained could be considered as representative values in free-living nestling peregrine falcons.     

Changes in serum protein profile of healthy adult Nigerians after three decades.

Serum protein profile was recently determined in 55 healthy adult Nigerians and compared with values obtained 3 decades ago. The total serum protein levels were similar, but an increase in titre was recorded in albumin, alpha 1-globulin and beta-globulin. The gamma-globulin level fell markedly in the present study, suggesting improvement in health care and quality of life in the last 3 decades. Changes seen in the serum protein pattern of healthy adult Nigerians were within the normal range compared with values recorded in other Africans and Black Americans. These changes should be considered in the interpretation of laboratory results.     

Serum proteins of the free-ranging European bison

The paper presents the values of total protein (TP) and protein fractions in the serum of 57 European bisons from the Bia?owieza Primeval Forest, 47 of these animals being free-ranging and 10 living in enclosures. It was found that the amount of TP and amount of alpha 1- and gamma-globulin fractions are significantly lower in adult females than males. The amount of TP and gamma-globulin fraction increase significantly with age in all examined Europeans bisons, whereas the amount of alpha 1- and alpha 2-globulin fractions increase only in free-ranging animals.     

Formation and morphology of struvite and newberyite in aqueous solutions at 25 and 37 degrees C

The influence of the initial reactant concentrations (c(i)(Mg)tot = 5.0 x 10(6) to 5.0 x 10(-1) mol dm(-3), c(i)(P)tot = c(i)(NH4)tot = 1.0 x 10(-3) to 5.0 x 10(-1) mol dm(-3)) and temperature (25 and 37 degrees C) on the composition and morphology of the precipitates formed in the system MgCl2-NH4H2PO4-NaOH-H2O at initial pHi = 7.40 has been investigated. Precipitation diagrams are presented showing the concentration regions within which different morphologies of solid phase have been formed. The solid phases aged for 24 hours were characterized by means of optical microscopy, FT-IR spectrophotometry, X-ray diffractometry and thermogravimetry. It was found that struvite was a predominant phase formed within the concentration region examined and newberyite was obtained only in the region where pH(24h) < 6.5. The influence of the initial pH on the formation and transformation of these two compounds were studied in the region 5.0 < or = pHi < or = 9.0 and the results are discussed.     

Agarose Gel Electrophoretic Fractionation of Serum Proteins in Adult Cattle. II. A Study Of Cows with Different Diseases

In 283 dairy cows suffering from internal disorders the serum proteins were studied by agarose gel electrophoresis supplemented by total protein and albumin determinations. The clinical diagnoses could be grouped according to protein pattern. Group 1 (abomasal displacement and traumatic muscle injury) did not appreciably affect the serum protein concentrations and represented primarily non-inflammatory diseases or diseases of non-infectious origin. Group 2 (leukosis) occupied an exceptional position, with heavy lowering of albumin unaccompanied by a corresponding rise of total globulin. The γ-globulin concentration was significantly lowered. Group 3 (acute traumatic peritonitis) represented an acute inflammatory process with increase chiefly of the α-globulins, while the γ-globulin concentration was normal. Group 4 (chronic traumatic peritonitis, summer mastitis, chronic subclinical mastitis, chronic laminitis and polyarthritis, urinary tract infection, abscess, and sub-acute-chronic pneumonia) comprised diseases chiefly of chronic inflammatory character and with infectious origin. Especially characteristic was the heavy rise of globulins in general and of γ-globulin in particular. In most cases there was a large increase also of the α- and p-globulin fractions.     

Water proton relaxation rate enhancements and association constants for Mn(II) to serum proteins determined by NMR T1 measurements

The water proton relaxation rate enhancement of Mn(II) bound to bovine serum albumin (BSA) and the association constant for manganese to BSA have already been determined, but such determinations have not been done for human serum albumin (HSA) and other human serum proteins and also for human serum. In this work, NMR T1 values in aqueous solutions of serum proteins and serum were measured versus increasing concentration of Mn(II). Proton relaxation rate enhancements (epsilon*) caused by different manganese concentrations were determined for each solution and 1/epsilon* was fitted against concentrations of Mn(II). Proton relaxation rate enhancements (epsilonb) of Mn(II) bound to albumin, gamma-globulin, (alpha+beta)-globulins and serum were found to be 13.69, 3.09, 8.62, and 10.87, respectively. Free and bound manganese fractions, resulted from each addition of Mn(II) to the sample, were determined by using corresponding (epsilon*) and the epsilonb values. Association constants for Mn(II) to HSA and gamma-globulin were calculated as 1.84 x 10(4) M(-1) and 2.35 x 10(4) M(-1), respectively. Present data suggest that the proton relaxation rate enhancement of Mn(II) in serum is caused by Mn(II) bound to various serum constituents. Data also suggest that association constants for Mn(II) to gamma-globulin are nearly the same as that to HSA.     

Segmental barrier properties of the pulmonary microvascular bed.

We determined liquid flux across single pulmonary microvessels of dog, ferret, and rat by our split-drop technique (J. Appl. Physiol. 64: 2562-2567, 1988). Data are reported from 58 lungs excised under halothane or pentobarbital sodium anesthesia and then blood perfused. We stopped blood flow at known vascular pressures and then micropunctured microvessels to inject oil, which we split with albumin solution. From measurements of vessel diameter and split oil drop length, we calculated Jv, the liquid transport rate per unit surface area [x 10(-6) ml/(cm2.s)]. At constant vascular pressure, Jv was not significantly different after different periods of oil-endothelium contact and at different sites within a single vessel. From measurements of Jv at different vascular pressures, we determined Lp, the hydraulic conductivity [x 10(-7) ml/(cm2.s.cmH2O)], and Pzf, the zero filtration pressure. From determinations of Pzf at different albumin concentrations, we quantified sigma alb, the albumin reflection coefficient. Lp and Pzf did not differ among venules of the same lung. However, in venules, Lp was 40% higher and sigma alb 25% lower than in arterioles (P less than 0.01). We conclude that 1) micropuncture procedures incidental to our split-drop technique do not progressively deteriorate the experimental microvessel and 2) in lung, permeability is higher in venules than in arterioles.     

Normal biochemistry values in baboons (Papio C. cinocephalus).

1. Analysis of biochemical parameters was carried out on material pooled from 10 baboons, adult male and female, of the Papio c. cinocephalus strain. 2. The values determined were for the common metabolites and enzymes utilized for specific studies in general metabolism (urea, glucose, cholesterol, phospholipids, bilirubin, uric acid, creatinine, alkaline phosphatase, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactic dehydrogenase, leucine amino peptidase, gamma glutamyl transpeptidase, total creatine kinase and amylase). Total protein and its fractions (ratio albumin/globulin, albumin, alpha 1-globulin and alpha 2-globulin) were also determined. 3. Results of these studies were compared with values for normal human adults. 4. The differences obtained between the respective ranges can be explained by the different physiology of the two species and, possibly, may also be caused by the use of human substrates in the enzyme assays. 5. Through the determination of normal values for the baboon, this study provides values for this species as an experimental animal in biomedical research.     

Experimental determination of net protein charge, [A]tot, and Ka of nonvolatile buffers in bird plasma.

The quantitative mechanistic acid-base approach to clinical assessment of acid-base status requires species-specific values for [A]tot (the total concentration of nonvolatile buffers in plasma) and Ka (the effective dissociation constant for weak acids in plasma). The aim of this study was to determine [A]tot and Ka values for plasma in domestic pigeons. Plasma from 12 healthy commercial domestic pigeons was tonometered with 20% CO2 at 37 degrees C. Plasma pH, Pco2, and plasma concentrations of strong cations (Na, K, Ca), strong anions (Cl, L-lactate), and nonvolatile buffer ions (total protein, albumin, phosphate) were measured over a pH range of 6.8-7.7. Strong ion difference (SID) (SID5=Na+K+Ca-Cl-lactate) was used to calculate [A]tot and Ka from the measured pH and Pco2 and SID5. Mean (+/-SD) values for bird plasma were as follows: [A]tot=7.76+/-2.15 mmol/l (equivalent to 0.32 mmol/g of total protein, 0.51 mmol/g of albumin, 0.23 mmol/g of total solids); Ka=2.15+/-1.15x10(-7); and pKa=6.67. The net protein charge at normal pH (7.43) was estimated to be 6 meq/l; this value indicates that pigeon plasma has a much lower anion gap value than mammals after adjusting for high mean L-lactate concentrations induced by restraint during blood sampling. This finding indicates that plasma proteins in pigeons have a much lower net anion charge than mammalian plasma protein. An incidental finding was that total protein concentration measured by a multianalyzer system was consistently lower than the value for total solids measured by refractometer.     

STIMULATION OF SYNAPTOSOMAL PROLINE UPTAKE BY SERUM ALBUMIN

Abstract— The accumulation of proline by synaptosomal fractions from adult brain cortices is strongly stimulated by serum albumin. Albumin at a concentration of 1 mg/ml increased the recovery of proline from the fraction by approx 2-fold. This effect was not due to any contaminant but rather due to the albumin molecule itself. Several other proteins tested failed to stimulate synaptosomal accumulation of proline, indicating that the stimulation due to albumin is not a general protein effect. The possibility that albumin stimulates synaptosomal proline accumulation by virtue of its fatty acid binding property is discussed.     

Some Effects of Protein Deficiency in Young Growing Pigs

No significant differences between the serum protein concentrations of the totally starved pigs and control animals given a milk protein supplemented diet were found. Serum lipid levels rose in the totally starved group. During the first 42 days of the experimental period sharp falls in serum protein concentrations were noted in the protein deprived pigs. The losses were greatest in the albumin and two β-globulin fractions, γ-globulin levels rose but at a much slower rate than in the control group. In the pigs which were refed a protein containing diet during the second 42 day period, the concentrations of all the serum protein fractions except for γ-globulin had reached control levels by the end of the investigation. The pigs which continued with the protein free diet showed further losses of albumin, a reduction in some α-globulin fractions and a cessation of net γ-globulin synthesis. The trends observed by measurements of protein bound carbohydrate were similar to those obtained from polypeptide determinations indicating quantitative rather than qualitative changes in the serum proteins. The reduction in the serum lipid concentrations of the pigs undergoing protein starvation was largely accounted for by losses of α- and β-lipoproteins. The protein deprived pigs maintained their initial body weight, while a continuous fall in weight was found in the totally starved group and a continuous increase in the control pigs.     

Competitive interactions of serum protein fractions during complex formation with polycations

Interaction of gamma-globulin with quaternized poly-4-vinylpyridine in water solutions at pH 7 has been studied. Formation of soluble stable cooperative complexes has been observed in a wide range of component ratios. Protein globules are distributed unevenly between adsorbing polycations. Soluble complexes are rod-like particles assembled from the globules which are stabilized by polycation chains. Complex formation in the system gamma-G + PE is similar to that in the system BSA + PE. Competitive interaction of serum protein fractions was studied at the interacting with polycation. It has been shown that selectivity at binding protein fractions is observed in both artificially prepared systems (BSA + gamma-G, beta1-G + gamma-G, BSA + gamma-G + beta1-G), and in serum and whole blood. In those ratios where uneven distribution of protein molecules is observed the soluble complexes protein-PE are formed by separate distribution of individual proteins at the matrix. Decrease of PE concentration in the systems results in the formation of a soluble complex of mixed composition. When an insoluble complex is formed in the system serum-PE selective sorbtion of beta 2-globulin fractions is observed. The reasons for the selective sorbtion of various protein fractions are described, structural models of the soluble complexes protein-PE are suggested.     

Stimulatory effect of prostaglandin F(2alpha) on Na-dependent phosphate transport in osteoblast-like cells

Prostaglandin F(2alpha) (PGF(2alpha)) has been reported to activate protein kinase C (PKC) through both phospholipase (PL) C and D, resulting in the proliferation of osteoblast-like cells. In addition, it has also been reported that Erk mitogen-activated protein kinase is also involved in the mechanism of PGF(2alpha)-induced proliferation of these cells. Recently, we have reported that several growth factors stimulate Na-dependent phosphate transport (Pi transport) activity of osteoblast-like cells, which has been recognized to play an important role in their mineralization. In the present study, we investigated the effect of PGF(2alpha) on Pi transport in MC3T3-E1 osteoblast-like cells. PGF(2alpha) stimulated Na-dependent Pi transport dose dependently in the range between 1nM and 10 micro M in MC3T3-E1 cells. The effect was time dependent up to 24h. Kinetic analysis revealed that PGF(2alpha) induces newly synthesized Pi transporter. Pretreatment with actinomycin D and cycloheximide suppressed PGF(2alpha)-induced enhancement of Pi transport. Combined effect of PMA and PGF(2alpha) was not additive in Pi transport. Calphostin C, a PKC inhibitor, dose-dependently suppressed Pi transport induced by PGF(2alpha). On the contrary, U0126, which inhibits an upstream kinase of Erk (MEK), did not affect PGF(2alpha)-induced enhancement of Pi transport. In conclusion, PGF(2alpha) stimulates Pi transport through activation of PKC in osteoblast-like cells.     

Immunochemical study of the specific antigens of human amniotic fluid]

Four specific antigens (trophoblastic beta 1-globulin, placental lactogen, alpha 1- and alpha 2-globulins of human placenta) were identified using antisera to the native amniotic fluid. Five antigens with the mobility of prealbumins, alpha 1-globulins, alpha 2-globulins and beta 2-globulins which bear no resemblance with the previously studied antigens were identified using antisera to the acid fraction of the amniotic fluid. Both the prealbumins and alpha 2-globulin were found in the blood serum of foetuses of different age and of newborn infants; these proteins were absent from the blood serum of pregnant women and donors. They received the names of embryonic prealbumine 1, embryonic prealbumine 2 and embryonic alpha 2-globulin. The protein with the mobility of alpha 1-globulins was found in the amniotic fluid of foetuses and in the blood serum of pregnant women only and received the name of amniotic alpha 1-globulin. The concentration of the antigens in question was studied in the developing foetuses and in the blood serum of pregnant women at different stages of pregnancy.     

Binding of 1alpha,25-dihydroxyvitamin D(3) to annexin II: effect of vitamin D metabolites and calcium

We have recently reported that annexin II serves as a membrane receptor for 1alpha,25-(OH)(2)D(3) and mediates the rapid effect of the hormone on intracellular calcium. The purpose of these studies was to characterize the binding of the hormone to annexin II, determine the specificity of binding, and assess the effect of calcium on binding. The binding of [(14)C]-1alpha,25-(OH)(2)D(3) bromoacetate to purified annexin II was inhibited by 1alpha, 25-(OH)(2)D(3) in a concentration-dependent manner. Binding of the radiolabeled ligand to annexin II was markedly diminished by 1alpha, 25-(OH)(2)D(3) at 24 microM, 18 microM, and 12 microM and blunted by 6 microM and 3 microM. At a concentration of 12 microM, 1beta, 25-(OH)(2)D(3) also diminished the binding of [(14)C]-1alpha, 25-(OH)(2)D(3) bromoacetate to annexin II, but cholecalciferol, 25-(OH)D(3), and 24,25-(OH)(2)D(3) did not. Saturation analyses of the binding of [(3)H]-1alpha,25-(OH)(2)D(3) to purified annexin II showed a K(D) of 5.5 x 10(-9) M, whereas [(3)H]-1beta,25-(OH)(2)D(3) exhibited a K(D) of 6.0 x 10(-9) M. Calcium, which binds to the carboxy terminal domain of annexin II, had a concentration-dependent effect on [(14)C]-1alpha,25-(OH)(2)D(3) bromoacetate binding to annexin II, with 600 nM calcium being able to inhibit binding of the radiolabeled analog. The inhibitory effect of calcium was prevented by EDTA. Homocysteine, which binds to the amino terminal domain of annexin II, had no effect on the binding of the bromoacetate analog to the protein. The data indicate that 1alpha,25-(OH)(2)D(3) binding to annexin II is specific and suggest that the binding site may be located on the carboxy terminal domain of the protein. The ability of 1beta,25-(OH)(2)D(3) to inhibit the binding of [(14)C]-1alpha, 25(OH)(2)D(3) bromoacetate to annexin II provides a biochemical explanation for the ability of the 1beta-epimer to inhibit the rapid actions of the hormone in vitro.     

Prolactin directly enhanced Na+/K+- and Ca2+-ATPase activities in the duodenum of female rats

Prolactin has recently been shown to directly stimulate 2 components of the active duodenal calcium transport in female rats, i.e., solvent drag-induced and transcellular-active calcium transport. Since the basolateral Na(+)/K(+)- and Ca(2+)-ATPases, respectively, play important roles in these 2 transport mechanisms, the present study aimed to examine the direct actions of prolactin on the activities of both transporters in sexually mature female Wistar rats. The results showed that 200, 400, and 800 ng/mL prolactin produced a significant increase in the total ATPase activity of duodenal crude homogenate in a dose-dependent manner within 60 min (i.e., from a control value of 1.53 +/- 0.13 to 2.29 +/- 0.21 (p < 0.05), 2.68 +/- 0.19 (p < 0.01), and 3.92 +/- 0.33 (p < 0.001) micromol Pi x (mg protein)(-1) x min(-1), respectively). Activity of Na+/K+-ATPase was increased by 800 ng/mL prolactin from 0.17 +/- 0.03 to 1.18 +/- 0.29 micromol Pi x (mg protein)(-1) x min(-1) (p < 0.01). Prolactin at doses of 400 and 600 ng/mL also significantly increased the activities of Ca(2+)-ATPase in crude homogenate from a control value of 0.84 +/- 0.03 to 1.75 +/- 0.29 (p < 0.05), and 2.30 +/- 0.37 (p < 0.001) micromol Pi x (mg protein)(-1) x min(-1). When the crude homogenate was purified for the basolateral membrane, the Na(+)/K(+)-ATPase activities were elevated 10-fold. In the purified homogenate, 800 ng/mL prolactin increased Na(+)/K(+)-ATPase activity from 1.79 +/- 0.38 to 2.63 +/- 0.44 micromol Pi x (mg protein)(-1) x min(-1) (p < 0.05), and Ca(2+)-ATPase activity from 0.08 +/- 0.14 to 2.03 +/- 0.23 micromol Pi x (mg protein)(-1) x min-1 (p < 0.001). Because the apical calcium entry was the first important step for the transcellular active calcium transport, the brush border calcium uptake was also investigated in this study. We found that, 8 min after being directly exposed to 800 ng/mL prolactin, the brush border calcium uptake into the duodenal epithelial cells was increased from 0.31 +/- 0.02 to 0.80 +/- 0.28 nmol x (mg protein)(-1) (p < 0.05). It was concluded that prolactin directly and rapidly enhanced the brush border calcium uptake as well as the activities of the basolateral Na(+)/K(+)- and Ca(2+)-ATPases in the duodenal epithelium of female rats. These findings explained the mechanisms by which prolactin stimulated duodenal active calcium absorption.