Α-globulin Fraction Research Articles

EFFECTS OF INANITION, SEMISTARVATION, AND PROTEIN-FREE DIETS ON SERUM PROTEINS.

The effects of inanition, semistarvation, and protein-free diets on the partition of serum proteins in adult, male, Sprague-Dawley rats was investigated following weight losses of 12, 26, and 33%. The quantitative changes in the serum protein fractions under the various types of nutritional stress showed significant differences. The albumin, α1- and γ-globulin fractions decreased significantly in the rats on the starvation regimen, while the semistarvation diet caused major alterations in the α1- and ß-globulins. Feeding a protein-free diet elicited the greatest decline in total serum proteins, reflecting the changes in the albumin, α1-, and α2-globulin fractions. The conclusion was drawn that the electrophoretic fractions of serum do not respond in a uniform and nonspecific manner to diverse nutritional stresses.

ISOLATION OF AN EASILY PRECIPITABLE ALPHA1-GLYCOPROTEIN OF HUMAN SERUM.

IN the course of investigations concerning the purification of human serum haptoglobin1 we found in the α1-globulin fraction obtained by preparative zone electrophoresis at pH. 8.6 a glycoprotein hitherto not described. Because of its unusual ability to precipitate at relatively low concentrations of neutral salts and ethanol we propose to characterize this α1-glycoprotein by the term ‘easily precipitable’.

Serum Glycoprotein Levels During Wound Tissue Regeneration

Abstract The level of protein-bound hexoses was compared in several fractions of electrophoretically separated globulins in the sera of normal and wounded rats. Changes in the concentration of protein components which were not apparent from the amount of total protein of different fractions could be identified. Although wounding resulted in no change from the normal protein level of one α-globulin fraction, the protein-bound hexoses decreased significantly below the normal. In another α-globulin fraction from wounded animals, a parallel increase in both protein and hexose above the normal was observed. The total proteins in the β-globulin fraction were not affected by wounding, even though there was an appreciable increase in hexose in the wounded over the normal level. In spite of the marked decrease in the amount of protein in the γ-globulins of wounded rats, no significant difference in the hexoses associated with this fraction was observed. Other globulin fractions were unchanged during wound tissue regeneration.

Changes in the Serum Protein Levels During Wound Tissue Regeneration

Abstract The effect of wounding and formation of regenerating wound tissue on the level of the serum proteins was studied in rats on a protein-free diet. It was found that the level of the globulin component having the third greatest electrophoretic mobility, probably part of the α2-globulin fraction, rose about 60% within 3 days after wounding and did not begin to return to normal until about 10 days later. Concomitantly, the level of the globulin with least mobility, presumably the -globulin component, decreased about 35% and also began to return to normal within 2 weeks after wounding. Varying the rate of healing, by the addition of protein or methionine to the diet, had no effect on the changes in the serum globulin levels. The newly formed α2-globulins were shown to be distinct from fetuin.

Production of embryonal alpha-globulin by transplantable mouse hepatomas.

It was previously reported that one strain of mouse hepatoma (strain XXIIa) contained an antigenic substance which was absent from liver, spleen, kidney, lung or serum of normal adult mice (1, 13). This antigen was isolated in an immunologically pure state (2, 3). It reacted only with anti-hepatoma sera and did not precipitate with antisera against liver, spleen, kidney or against normal adult mouse sera. The present work, however, demonstrates that this antigen is a normal constituent of the embryo and of newborn mouse serum, and that it disappears from the serum as the mice become older. It also shows that the same antigen reappears in the serum of adult hepatoma-bearing mice as a result of its synthesis in the tumor tissue followed by its secretion into the blood. Judged by its electrophoretic mobility this antigen is related to the α-globulin fraction of mouse serum.

Age changes in plasma proteins of the Mongolian gerbil.

The effect of animal age on plasma protein composition was studied in the Mongolian gerbil, Meriones unguiculatus. Values for plasma protein fractions separated by electrophoresis were obtained from bromophenol blue-stained paper strips by elution and by scanning techniques, both of which yielded similar results. Total plasma proteins increased with age from 1 day to 28 months. Maximum albumin concentrations were attained at 3 months and maximum (adult) γ-globulin-fibrinogen concentrations at 28 months. A fast α2-globulin fraction was found in plasma from the youngest age group (1–3 days) but absent in older groups. No sex-related differences in plasma proteins were found for gerbils at 3 and 6 months. Age-related changes in gerbil plasma proteins are similar to changes found during the life span of many other mammalian species.

DECREASED ANTICOAGULANT TOLERANCE DURING METHANDROSTENOLONE THERAPY.

Administration of methandrostenolont (δ1 -17α-methyltestosterone) in a dosage of 10 mg a day to patients on maintenance therapy with warfarin sodium or phenylindanedione was found to produce, within two weeks, a significant decrease in the anticoagulant requirement.The response to a single dose of 20 mg of warfarin sodium in healthy subjects was found to be increased when the test subjects had taken methandrostenolone 10 mg daily for 10 days. Liver function studies in healthy subjects, who received methandrostenolone in a similar dosage for 10 days, revealed a significant increase of the α2-globulin fraction, a significant decrease of alkaline-phosphatase activity, and a significant decrease of bromsulfalein clearance.

Localization of serum leucine aminopeptidase activity by paper electrophoresis.

The electrophoretic behavior of serum leucine aminopeptidase (LAP) in normal individuals, in pregnancy, and in patients with hepatobiliary disease was investigated with paper electrophoresis technics. In normal serum LAP migrated as a single component in the α1-globulin fraction. In patients with elevated serum LAP activity due to hepatobiliary or pancreatic disease supplementary peaks of enzymatic activity were demonstrable in other protein fractions. Thus, in jaundiced patients with carcinoma of the head of the pancreas, liver metastases, biliary atresia, acute hepatitis, and drug-induced jaundice, additional activity was observed in the β-globulin fraction. In some patients with hepatitis additional activity was observed in the α2-globulin as well as β-globulin. In common duct stone additional activity in the albumin fraction was a consistent finding. In early pregnancy the electrophoretic pattern of LAP was similar to that observed in normal serum, whereas during the third trimester LAP activity was demonstrable only in the α2-globulin.

The nature of pig antibodies against Brucella suis.

(1) In immune pig serum, agglutinins againstBrucella suis were found to be distributed in the γ-and β-globulin fraction. Immunochemical determination also showed that gamma globulin antigen is present in the β-globulin fraction and some even in the α-globulin fraction. (2) It was demonstrated by two ultra-centrifugation methods that agglutinins againstBrucella suis belong both to 7 S type and 18 S type. The individual types were separated by chromatography. (3) Fractions containing antibodies were prepared from immune serum by precipitation with alcohol, sodium sulphate and rivanol. In contrast to preparation by zone electrophoresis, none of these methods succeeded in including all the serum antibody activity in the product. (4) The stability of the antibody activity of two different molecular types of anti-brucellar antibodies differs at pH 2.5–3. The activity of 18 S antibody is largely destroyed under these conditions, while that of the 7 S type maintained.

Fractionation of plasma parathyroid hormone activity.

Components of rat and human plasma obtained by ammonium sulfate and cold ethanol fractionation procedures have been assayed for their calciummobilizing activity in parathyroidectomized rats. The determinations were made on fractions of rat plasma to which parathyroid extract (Lilly) was added either in vitro or in vivo. Similar experiments were carried out on fractions of human plasma with and without added parathyroid extract. Evidence is presented to suggest that endogenous rat plasma parathyroid hormone activity is normally associated with the α-globulin fraction. Cold ethanol fractionation of normal human plasma and of human plasma to which had been added parathyroid extract, resulted in localization of hormonal activity in the beta and gamma globulin fraction. The normal level of circulating parathyroid hormone activity in human plasma has been estimated to be of the order of 40 U.S.P. units per 100 ml.

Nature of thyroxine-binding globulin in human serum.

THE term thyroxine-binding globulin is often used to denote a serum protein fraction with a mobility intermediate between those of α1 - and α2-globulins in paper or moving-boundary electrophoresis in veronal buffer, pH 8.6, ionic strength 0.05–0.10 (ref. 1). However, with the introduction of supporting media such as agar and starch gels and the use of different buffers, ionic strength or pH for paper electrophoresis, binding of thyroid hormones has also been detected in the ‘prealbumin’ zone2. Although the proteins of the ‘prealbumin’ zone obtained under different conditions are not likely to be identical, a strong thyroxine-binding property was detected in human serum prealbumin rich in tryptophan3, as obtained from Prof. Schultze4. From chemical and immunological observations I concluded that the tryptophanrich prealbumin, present as a complex in the α-globulin fraction, might be responsible for much of thyroxine-binding in human serum5. Since then, an α-globulin-type of thyroxine-binding protein has been obtained in this laboratory (unpublished results); the nature of this protein does not support the hypothesis that the tryptophan-rich prealbumin is involved in binding thyroxine. One possible explanation for this discrepancy is that Schultze's prealbumin preparation contained traces (1–2 per cent) of thyroxine-binding globulin which were responsible for the production of antibodies to it present in antisera against this prealbumin. In fact, multiple precipitation lines were observed in immunological studies using rabbit anti-human prealbumin serum5. The experiments to be described support this explanation and throw further light on the nature of the thyroxine-binding globulin in human serum.

Effect of induced hypoxemia on serum protein fractions. A clinical pathophysiological study

Concerning the metabolism of proteins in hypoxemia, in 1920, Graubitz, Tscherkes and Melnikova found increases in the total nitrogen and NH3 of the urine when carbonmonoxide hypoxemia was induced in animals, which indicated an increase in protein breakdown. However, no report has appeared in the literature on the changes in human serum proteins under acute induced hypoxemia. Therefore, the author subjected normal individuals and patients with various diseases to low oxygen respiration with the intention of studying its influence on the serum proteins. Methods and Subjects Human subjects in a resting and fasting state were made to inhale a 10% oxygen gas mixture for 15 minutes. Blood samples were obtained from. the femoral artery before and at the end of the inhalation. Also, in some cases, blood of the hepatic vein was obtained by the hepatic catheterization method. Serum protein fractionation of these samples was done by paper electrophoresis. The results obtained were analysed and their relations to the data of blood gas and carbohydrate metabolites obtained by coworkers, were studied. A total of 27 subjects were studied, which included healthy individuals (16 cases) and patients with mild or moderate pulmonary tuberculosis, hypertension and arteriosclerosis. Hepatic catheterization was per-formed on 14 cases which included healthy individuals (10 cases) and patients with pulmonary tuberculosis, gastric ulcer or mild cholecystopathy. Low oxygen administration was done simultaneously on 6 of these cases. RESULTS I. SYSTEMIC ARTERIAL BLOOD 1) Changes in the serum protein fractions : The low oxygen inhalation caused a significant decrease of the albumin fraction and a significant increase of the aα1 globulin fraction. All globulin fractions showed negative correlations with the albumin fraction before the low oxygen inhalation and the correlations were also seen after the inhalation in all fractions except the α1 globulin fraction. In general, the albumin fraction was most easily affected by the hypoxemia and the major change of serum proteins caused by the low oxygen respiration was the decrease in the albumin.

Electrophoretical Studies on Soluble Proteins from the Tissue of Organs

For the purpose of electrophoretically studying the relation between the general and the local abnormal metabolism in malignant tumor, changes in the levels of proteins, glycoprotein and protein-bound polysaccharide from the tissue were followed up in comparison with findings of serum proteins in the tissue of the stomach removed from cases of gastric cancer and ulcer and also in the tissue of the stomach excised from gastric sarcoma-implanted and Yoshida sarcoma-implanted rats. The following conclusions were thus reached:1. The characteristic findings of proteins from the tissue of cases of tumor were a very remarkable decrease in the albumin fraction, an increase in the α1-globulin fraction and the α1-glycoprotein fraction and a significant increase in the protein-bound polysaccharide level. Though these findings showed the same tendency as observed in the findings of serum proteins, they appeared earlier than the latter and were considered to be characteristic of the tissue from cases of malignant tumor.2. This pronounced decrease in the albumin fraction was a main factor in a decrease in the serum albumin and total protein levels.3. An increase in the protein-bound polysaccharide level seemed to be chiefly responsible for an increase in the serum protein-bound polysaccharide level.4. An increase in the α2-globulin fraction and the α2-glycoprotein fraction which was observed in serum was a finding which was equally met with in cases of phthisis where proliferation or inflammation or degeneration of cells occurred. The increase seemed to be parallel to the extent of changes in these phenomena.5. In other words, the foregoing findings can be regarded as indicating (1) the general and local abnormal metabolisms in the cancer-carrying organism, (2) the close relationship between both metabolisms and (3) the process in which local changes in malignant tumor gradually pass into findings of serum proteins.

Metabolic Abnormality of Serum in Diabetic Disease

With a view to studying abnormalities of metabolism in diabetes, measurement was paperelectrophoretically made of serum protein, compound protein, cholesterol, esterified cholesterol and protein-bound polysaccahride levels in diabetic patients and rabbits with alloxan-induced diabetes.Observation was made on the diabetic patients in order to detect the presence of signs of arteriosclerosis and also to seek a relation between sick periods. Comparison of the results between rabbits with alloxan-induced diabetes and diabetic patients was made with the following results:1. In diabetic patients the total protein level and albumin fractions generally showed a decrease. The extent of that decrease was a little greater in those who showed signs of arteriosclerosis. Those patients with signs of arteriosclerosis howed a remarkable increase in the α2-globulin fraction. The longer was the period of illness, the greater was a decrease in the albumin fraction and an increase in the α2- and β-globulin fractions. These changes had the same tendency, though different in extent, in rabbits with alloxan-induced arteriosclerosis.2. There was a decrease in the α-lipoprotein fraction and an increase in the β-lipoprotein fraction. Both the cholesterol and esterified cholesterol levels showed an increase. That increase was marked in patients with signs of arteriosclerosis. An insignificant decrease in the ester/cholesterol ratio occurred in these patients. The longer was the sick period, the greater was that increase. The same tendency was observed in rabbits with alloxan-induced arteriosclerosis. In this case, the relationship between the β-lipoprotein fraction and the cholesterol level was found in diabetic patients and not in rabbits with alloxan-induced arteriosclerosis.3. Glycoprotein fractions (albumin fractions and α- and β-fractions) which had a great electrophoretic mobility exhibited an increase in general, and an increase in the α2-glycoprotein fraction in particular was striking. There was an increase in the protein-bound polysaccahride level along with that increase. The longer was the period of sickness, the greater was the increase in the protein-bound polysaccahride level. On the contrary, rabbits with alloxan-induced arteriosclerosis showed a decrease in the α-glycoprotein fraction, but exhibited an increase in the protein-bound polysaccharide level.

Clinical and Biochemical Studies on Serum Protein and Compound Protein

Protein, lipoprotein (Lp) and gfycoprotein (Gp) were studied electrophoretically in liver disease and experimental liver impairment which was induced in rabbits. Total protein (T.p), cholesterol (Ch) and protein-bound polysaccharide (PBP) were also studied.Rabbits used in this experiment were injected with Indian ink and CCl4. Liver diseases studied were hepatitis, liver cancer and liver cirrhosis.1. Changes, which were related to the extent or progress of liver impairment when the rabbits were treated with Indian ink, were a decrease in the T. p and albumin levels and a corresponding increase in the γ-globulin and α1-globulin fractions. In the case of the Lp level, there was an increase in the total Ch level and a decrease in the β- and γ-Lp fractions. The esterified cholesterol level was in parallel to the total cholesterol level. In the case of the Gp level, the PBP level did not undergo so great changes. A tendency for the α1-Gp fraction to increase was interesting.2. In the case of CCl4 injection, changes which were related to the extent of liver impairment were that the T. p level showed an increase after a temporary decrease and that the albumin fraction followed opposite courses to the T. p level. The γ-globulin fraction was corresponding to the albumin fraction and showed almost the same tendency as observed in the case of Indian ink. An increase in the α2-globulin fraction was observed in patients who had died. In the case of the Lp level, the total and esterified cholesterol levels showed a sudden decrease after the injection, and the esterified cholesterol/total cholesterol ratio also exhibited a decrease. On the other hand, β-Lp fraction increased after it decreased. In the case of the Gp level, the PBP level did not undergo so great changes. It showed a temporary increase in some cases, but its decreasing tendency was noticed. A tendency forthe α1-Gp fraction to increase was observed.3. In clinical experiments a decrease in the T. p level and the albumin fraction as well as an increase in the γ-globulin fraction was perceived in many instances. The decreasing rate of the T. p level was the smallest in liver cancer, followed by liver cirrhosis and hepatitis. The decreasing rate of the albumin fraction was greater in liver cirrhosis than in liver cancer, and the α2-globulin fraction showed a high value in liver cancer. In the case of hepatitis, a difference according to the intensity of its symptoms was reflected in the T. p level. As foa the Lp level, an increase in the total cholesterol and esterified cholesterol levels and the β-Lp fraction occurred in the advanced stage of icterus in hepatitis, while there was a decrease in these levels in cases of great liver function impairment. The total cholesterol level was high in cases of liver cirrhosis which followed favorable courses, whereasit was low in cases which followed unfavorable courses ann especially in cases of death. It showed a decrease in the progressive stage of liver, cancer. In the case of the Gp level, the PBP level was high in cases of hepatitis where jaundice was marked, while it was low in cases where liver function impairment was great. In the stadium acme of hepatitis, there was an increase in the α1-Gp fraction. In liver cancer, the PBP level and the α1- and α2-Gp fractions showed an increase and these exhibited a greater increase along with the progress of this disease. The PBP level was low in cases of liver cirrhosis which followed unfavorable courses, while it was high in cases Which followed favorable courses. The high α1- and α2-Gp fractions were reduced by medical treatment, whereas the low ones showed a tendency to be increased.

Electrophoretical Studies on Proteins from the Tissue of Internal Organs

Soluble protein from the liver tissue was studied by paper electrophoresis in an attempt to throw light on metabolic abnormalities in cancer from the standpoint of changes in tissue proteins from cancer-bearing organisms and the relationship between changes in the tissue and serum proteins. It was considered to serve the above purpose most properly to know changes in tissue proteins from the liver in metabolic abnormalities because the liver is an organ which plays a major part in intermediary metabolism.In part I, investigations were made into basic conditions, e. g., tissue collection, extracting solutions, and the effects of blood contained in samples, the effects of trituration time on the extraction of soluble proteins, the extracting time, the removal of turbidity from extracting solutions, the concentration of proteins in samples, etc. In part II, normal rats and ascites-induced rats were compared with Yoshida-sarcoma-implanted rats on the basis of the results obtained in part I.1. Fractions of protein and compound protein from saline-extracted liver tissues were not satisfactorily separated as those of serum protein were. The electrophoresis diagrams of the proteins were entirely different from those of serum protein.2. The amount of tissue protein from the liver showed a decrease in sarcoma-implanted rats. This decrease was due not to proliferation of the sarcoma, but to a decrease in fraction A corresponding to the albumin plus α1-globulin fraction of serum.3. The total PBP level of sarcoma-implanted rats decreased in the liver tissue, contrary to that in serum. Parallelism existed between the level and the tissue glycoprotein fraction A (corresponding to the albumin plus α1-globulin fraction of serum glycoprotein).4. Like in the case of serum, the liver cholesterol level showed an increase, and there was a relationship between both. Parallelism was also found between the level and the tissue lipoprotein fraction C.

Electrophoretic studies of serum proteins and glycoproteins during early stages of experimental tuberculosis.

SummaryThe systemic effects of acute experimental tuberculosis as reflected by alterations in concentration and distribution of the proteins and protein-bound carbohydrates of serum were investigated by filter paper electrophoresis. Electrophoretic analyses were performed on serum samples from groups of adult female guinea pigs sacrificed 8, 15, 19, 29, and 50 days following inoculation with 0.1 mg M. tuberculosis. Significant increases occurred in total serum glycoprotein levels and in the protein component of the α1-globulin fraction 8 days post-infection. A marked elevation of the polysaccharide moiety of the α2-globulin fraction and a significant decline in albumin protein was observed 15 days following infection. With the exception of significant increases in the components of the γ-globulin fraction in the latter stages of the study, major alterations in the other subfractions of serum had occurred by 15 days postinfection. It is suggested that the early alterations in electrophoretic patterns repre...

STUDIES ON A RTERIOSCLEROSIS WITH RESPECT TO ELASTASE-INHIBITOR IN THE SERUM

The writer has presented his studies on arteriosclerosis from an angle of elastase which is closely connected with the proliferation and destruction of elastin in the artery. Special attention has been given to an elasta e-inhibitor in the serum and the findings are as follows : 1. Optimal pH of elastase was from 8.4 to 8.9.2. The Ca-Salt inhibits elastase activity.3. Elastase-inhibitor exists in the serum of both men and rabbits. This material is for the most part found in both albumin fraction and α1-globulin fraction of the serum protein; it is unstable under heating and its activity is lost by heating (over60°C); this material has little to do with the quantity of the serum total protein, nor is it influenced by administering heparin in jection.4. In the case of the rabbits which were fed adeps lanae, the more serious arteriosclerosis is, the less serum elastase-inhibitor exists.5. Elastase-inhibitor in the group which shows serious sclerosis in eyeground exists less than in the group which shows no sclerosis in eyeground.6. The decline of elastase-inhibitor in the abnormal electrocardiographic group was stronger than in the normal group.7. The decline of serum elastase-inhibitor is seen in various diseases which were closely related to arteriosclerosis. And in the human aorta, the decline of elastase-inhibitor was greater where sclerosis was clear than in a normal part.8. From said findings. it is inferred that elastase-inhibitor plays a considerable part in clarifying cause of arterioselerosis.

Dietary Protein and Serum Electrophoretic Patterns of the Adult Rat.

SummaryThe effects of inanition, acute and chronic protein depletion and of repletion with several levels of protein on distribution of serum proteins in the adult rat have been investigated by filter paper electrophoresis. Decreases in the albumin, α1-, α2- and β-globulin fractions were observed following depletion regimens. Purified diets containing 17% or more of casein not only restored all values to normal, but elicited significant increases in total serum protein concentrations. A differential response of the α2-globulin fraction upon repletion occurred between groups previously subjected to acute and to chronic depletion. Significantly increased A/G ratios were found in groups repleted with a diet of natural products. Rate of depletion was found to exert a pronounced effect on hematocrit values.

Prealbumin as a complex in the alpha-globulin fraction in human serum.

Prealbumin as a complex in the alpha-globulin fraction in human serum.