Prealbumin Fraction Research Articles

PROTEIN ELECTROPHORESIS AND REFERENCE INTERVALS OF SERUM PROTEINS OF A CAPTIVE POPULATION OF ROTI ISLAND SNAKE-NECKED TURTLES (CHELODINA MCCORDI).

Protein electrophoresis (PEP) is an important tool in mammals to characterize specific dysproteinemias and detect acute and chronic inflammatory responses. In reptiles, PEP is the gold standard method for globulin fraction determination and albumin measurement. In this study, preliminary reference intervals were established for serum PEP in 22 clinically healthy adult Roti Island snake-necked turtles (Chelodina mccordi), a critically endangered species, kept in captivity and sampled over two monsoon seasons. The species has a prominent prealbumin fraction and β-globulins were the dominant globulin fraction. Significant differences between females and males were found in prealbumin (P < 0.01), albumin (P = 0.02), α1-globulin (P = 0.05) and γ-globulin (P = 0.01). Gravid females had significantly lower total protein (P < 0.01), prealbumin (P < 0.01), albumin (P < 0.01) and albumin:globulin ratio (P = 0.01). These preliminary reference intervals should aid in clinical investigation in this species as well as further research studies seeking to understand the application of PEP in reptilian species.

PLASMA PROTEIN ELECTROPHORESIS IN THE WHITE STORK (CICONIA CICONIA): AGREEMENT BETWEEN AGAROSE GEL VERSUS CAPILLARY ZONE METHODS AND DEVELOPMENT OF REFERENCE INTERVALS.

The white stork (Ciconia ciconia) is a ciconiiform species widely represented in zoological institutions. Plasma protein electrophoresis is widely used in avian patients for assessment of inflammatory conditions, but reference intervals for this testing modality are lacking for the white stork. The two main electrophoretic methods are agarose gel electrophoresis (AGE) and capillary zone electrophoresis (CZE). This study assessed fresh plasma samples of healthy adult white storks (n = 30). Statistical analyses were performed to evaluate agreement between AGE and CZE. Typical electrophoretic fractions were obtained from both methods (prealbumin, albumin, α1, α2, β, γ1, and γ2). The AGE and CZE methods were not equivalent for determining major electrophoretic fractions (except β-globulins) and albumin:globulin ratio on plasma samples. An additional prealbumin fraction was seen with CZE. Reference intervals were established for each method as the smallest n group was 27 individuals for a given value; most values had normal distribution, and robust or parametric methods were used on the data.

Changes of total protein and protein fractions in broiler chickens during the fattening period.

Background and Aim:Blood proteins in birds serve as an important indicator in the evaluation of health status and represent a basis in general biochemistry allowing the identification of metabolic alterations. The objective of this study was to evaluate the protein profile in broiler chickens extended by the concentrations of serum protein fractions at different periods of fattening.Materials and Methods:Into the evaluation, we included 24 clinically healthy Ross 308 line meat-type chickens at the age of 2 days. Blood samples were taken on day 4, 18, 32, and 46 of fattening always from six randomly selected chickens. Chickens were fed with a commercial starter, grower, and finisher feeds. The concentrations of total serum protein and protein fractions were evaluated.Results:Various significant changes in the proportion of the individual protein fractions were found during the observed period except for the beta-globulins in all protein fractions and the albumin/globulin (A/G) ratio. At the beginning of the fattening period, the relative concentrations of albumin, α1-globulins, and A/G ratio were significantly lower and the values of α2- and γ-globulins significantly higher (p<0.05). The values of pre-albumin fraction were found as a small band preceding the albumin fraction differed significantly between the different age groups of chickens (p<0.05). The total serum protein concentrations showed higher values in older broilers; the significantly highest mean value was recorded on day 32 of fattening.Conclusion:The results suggest that fattening and age of broilers influences not only the production patterns, metabolic processes, and lipid and mineral profile but also the parameters of protein profile. However, seeing that some contradictory data exist regarding the number and size of globulin fractions in chickens, further analyses are needed.

Plasma Protein Electrophoresis in Birds: Comparison of a Semiautomated Agarose Gel System With an Automated Capillary System

Plasma agarose gel electrophoresis (AGE) is recognized as a very reliable diagnostic tool in avian medicine. Within the last 10 years, new electrophoresis techniques such as capillary zone electrophoresis (CZE) have emerged in human laboratory medicine but have never been investigated in birds. To investigate the use of CZE in birds and to compare it with AGE, plasma samples from 30 roosters (Gallus gallus), 20 black kites (Milvus migrans), and 10 racing pigeons (Columba livia) were analyzed by both AGE and CZE. For the 3 species studied, values determined by AGE and CZE were well correlated for albumin and beta and gamma fractions whereas other values differed significantly. Values for alpha-3 fraction in the rooster, alpha-1 fraction in the black kite, and alpha fractions in the pigeon obtained by AGE were very well correlated with the prealbumin fraction values obtained by CZE. Repeatability and reproducibility appeared higher with CZE than with AGE. Although the interpretation of CZE electrophoresis patterns seems to produce results similar to those obtained with AGE, some proteins present in the alpha fraction measured with AGE migrated to the prealbumin fraction found with CZE. Although CZE requires the use of specific reference intervals and a much higher sample volume, this method has many advantages when compared with AGE, including better repeatability and reproducibility and higher analysis output.

An appraisal of blood-cerebrospinal fluid barrier dysfunction during the course of Guillain Barré syndrome

Elevated cerebrospinal fluid (CSF) total protein (TP) concentration (mainly due to a dysfunctional blood-CSF barrier (B-CSFB)) with normal cell count is a hallmark for the diagnosis of Guillain-Barriota syndrome (GBS). This work presents the evaluation of B-CSFB dysfunction with respect to the course, severity, and clinical features of GBS. A sample of CSF was collected on admission from 68 patients of both genders (15 children and 53 adults) diagnosed with GBS. A follow-up CSF sample was obtained approximately 15 days after admission. TP concentration was determined in the CSF and 7.5% polycrylamide gel electrophoresis was employed for serum and CSF protein fractioning. A low percentage of prealbumin fraction was considered a test of impaired B-CSFB. Elevated TP concentration and lower prealbumin were observed in almost 70% of the patients on admission, but this percentage was lower (52.4%) during the first week from onset of symptoms. Both variables were directly associated with the time of evolution of the disease and also with a greater clinical severity. Follow-up CSF studies showed higher CSF TP and lower prealbumin percentages, while deceased patients did not display this response pattern in the follow-up CSF. B-CSFB dysfunction was present in only half of the patients with GBS during the first week from onset and it was directly associated with progression and clinical severity; nevertheless, a low B-CSFB dysfunction response during follow-up was associated with a lethal outcome, suggesting it could also serve a 'protective' effect during regeneration.

Selenium toxicosis assessment ( in vivo and in vitro) and the protective role of vitamin B12 in male quail ( Coturnix Coturnix)

The present study was undertaken to elucidate the toxicity induced by sodium selenite in male quail through in vivo and in vitro studies and the role played by vitamin B12 in alleviating selenium toxicity. Administration of selenite orally for 1 month induced hepatic oxidative damage. Selenite decreased body weight gain and increased relative liver weight. Selenite reduced hemoglobin and iron concentrations and elevated total bilirubin concentration. Serum transaminases and alkaline phosphatase activities were increased in selenium-intoxicated quails. Total protein concentration was decreased associated with the appearance of prealbumin fraction, an increased γ-globulin and a decreased α- and β-globulins. The highest level of selenium was found in liver followed by kidney, testis, faeces and blood. Supplementation of vitamin B12 orally for 1 month simultaneously with selenite caused less marked biological alteration in the investigated parameters. In vitro study using isolated quail hepatocytes incubated with sodium selenite showed a dose-dependent response for toxicity markers. These results suggest that selenosis can be reduced by vitamin B12 supplementation.

Plasma Protein Electrophoresis of the Atlantic Loggerhead Sea Turtle, Carretta carretta

The objective of this study was to determine reference intervals for plasma protein fractions of normal appearing, wild Atlantic loggerhead sea turtles, Caretta caretta. Blood was collected into heparinized vacutainer tubes from the following groups of turtles: 1) ten adult males; 2) eleven adult females; 3) ten juve- nile males; and 4) ten juvenile females. Plasma was removed and total protein content of each sample was determined using the biuret method. Plasma proteins were separated using gel electrophoresis and scanned using a laser densitometer. Reference ranges for albumin, alpha, beta, and gamma globulins were established for age and gender classes and statistically analyzed. Significant differences were found between beta globu- lins of adult and juveniles and between juvenile males and females. A subgroup of turtles had electrophoretograms with beta-gamma bridging and a single adult male loggerhead had a prealbumin fraction; however, these subgroups of turtles were excluded from statistical analysis.

Avaliação do perfil eletroforético das proteínas séricas em matrizes pesadas (Gallus Gallus Domesticus) da linhagem Avian Farm

O objetivo do presente trabalho foi determinar o perfil eletroforético das proteínas séricas em matrizes pesadas (Gallus Gallus Domesticus) sadias, da linhagem Avian Farm. Foram utilizadas 15 matrizes com 63 semanas de idade, provenientes do município de Conchas, São Paulo. Utilizou-se o método de biureto para a obtenção dos valores da proteína sérica total e a separação das frações protéicas pela técnica da eletroforese em gel de agarose, e a leitura do filme realizada por densitometria em 520nM. Obteve-se um total de sete frações, sendo que a beta1 - globulina e beta2 - globulina encontradas não foram citadas pelos autores consultados na literatura. A fração pré-albumina foi identificada em apenas seis das 15 amostras examinadas. Observou-se em cinco matrizes a divisão da gama - globulina em duas frações, denominadas gama - 1 e gama - 2, de acordo com suas mobilidades eletroforéticas. A relação albumina/globulina (A/G) encontrada corrobora com os autores citados, demonstrando que esta diminui com o aumento da idade.

Effects of fasting and 3,3',4,4',5,5'-hexabromobiphenyl on plasma transport of thyroxine and retinol: fasting reverses elevation of retinol.

Male Wistar rats were injected ip with 0 or 20 mg/kg 3,3',4,4',5,5'-hexabromobiphenyl and blood samples were collected 1, 3, 6, 7, and 8 d later. At 8 d after the injection, serum retinol was increased 30% and serum thyroxine was decreased 26% relative to control values. These effects were apparently unrelated to transthyretin in that the biphenyl did not alter the proportion of thyroxine binding in vitro to the prealbumin fraction of serum proteins. Separate groups of control and HBBP-injected rats did not receive food on d 7 (i.e., 24-h fast) and d 8 after injection (i.e., 48-h fast). Fasting decreased the serum retinol and thyroxine concentrations as well as the proportion of thyroxine binding in vitro to the prealbumin fraction of serum. The decreases in retinol and thyroxine concentrations associated with fasting are therefore ascribed to a decrease in the concentration of transthyretin in circulation.

Cerebrospinal fluid proteins in muscular dystrophy patients

We analyzed and quantified the microcomponents of protein fractions in the cerebrospinal fluid of patients with various types of muscular dystrophy. The degenerative pattern is characterized by an increase in the prealbumin fraction and a decrease in the gamma-globulin fraction as shown in the Duchenne and congenital muscular dystrophy. The increase in CSF IgG, gamma-globulin fraction, and myelin basic protein is shown in the myotonic dystrophy. In addition to the lowness of IQ, and the abnormality of EEG and brain CT, abnormal CSF proteins obviously suggest the presence of CNS involvement in muscular dystrophy.

A hemolytic lipoprotein containing lysophosphatidylcholine produced in incubated mouse plasma

A factor capable of lysing erythrocytes arose in the plasma of various strains of mice when the plasma was incubated at 37°C. The hemolytic activity and the amount of lysophosphatidylcholine (lysoPC) in the plasma reached a maximum at 3 h of incubation. Both the hemolytic activity and the increment portion of lysoPC were recovered from the prealbumin fraction on agarose gel electrophoresis. Serum albumin, an inhibitor of the hemolytic action of lysoPC, inhibited the hemolytic action of the this factor. These and other results indicate that the hemolytic factor is a lipoprotein that contains lysoPC as an active site. The molecular weight of active fraction was distributed from 100000 to 200000. The finally purified fraction contained 65% protein composed of a polypeptide with a molecular weight of 25000. The lipid composition of this fraction was 12.4% lysoPC, 63.9% phosphatidylcholine (PC), 5.4% sphingomyelin, 2.1% triacylglycerols and 16.2% cholesterol. This hemolytic lipoprotein was also found in the prealbumin fraction when electrophoresis of plasma of the lysoPC-administered mice was carried out. However, such lipoprotein was not formed by simple addition of lysoPC to the nonincubated plasma.

Common antigenicity between a human thymic epithelial cell product and a thymus-dependent serum factor.

Specific immunofluorescence of human thymic epithelial cytoplasm was obtained with antibodies to supernatant of thymic epithelial cultures, and with anti-prealbumin antibodies. These antibodies also reacted with normal serum but not with serum from Di George patients. The data indicates that thymic epithelium and a component of the prealbumin fraction of normal serum share a common antigen believed to be thymic hormone.

Human prealbumin fraction: Effects on cell-mediated immunity and tumor rejection

The effect of human prealbumin fraction on allogenic cell-mediated immunity in primary sensitization cultures of murine spleen cells was studied by 3H-thymidine uptake and specific 51Cr release assays. Prealbumin caused a dose-dependent augmentation of these responses. Human serum albumin, bovine serum albumin, and calf-thymosin fraction 5 had little effect. Prealbumin was active when added on day 0 or 1 but not thereafter. Prealbumin added to effector cells from immunized mice did not change their lytic activity. Prealbumin, but not human serum albumin or thymosin fraction 5, augmented secondary cell-mediated immunity in culture after primary immunization in mice. A slow growing mammary tumor line, which originated as a spontaneous mammary tumor in a DBA/2a HaDD breeder mouse, initially grows in 100% of DBA/2J mice but is then rejected in 10–20% of them. When prealbumin (50 μg/day) was given subcutaneously for 2 weeks to DBA/2J mice and the tumor implanted 2 weeks later, 78% of the mice rejected the tumor and were then resistant to a rechallenge.

Protein composition in single follicles, homogenates and fine-needle aspiration biopsies from normal and diseased human thyroid

The protein composition of the thyroid colloid was analysed by microgel electrophoresis and densitometry in 41 euthyroid patients. The colloid samples were obtained from single follicles by micropuncture, from homogenates of microbiopsies or from aspiration biopsies. Fourteen of the patients had morphologically normal thyroid tissue, 18 had atoxic nodular goitre and 19 of the patients had atoxic adenoma. Ten of the patients with nodular goitre had prior to the investigation received lithium therapy for psychiatric disorders. The main component of the thyroid colloid was 19S thyroglobulin (TG), but larger iodoproteins (S-TG) and smaller protein fractions, an albumin-like protein and a pre-albumin fraction, were also present in varying relative amounts. Analyses of homogenates of microbiopsies from normal thyroid tissue demonstrated the same protein composition as observed in single follicles. In colloid samples from atoxic nodular goitre the lighter protein fractions were absent in most of the samples. Analyses of homogenates or aspiration biopsies could not demonstrate this alteration in the protein composition in nodular goitre. Lithium therapy resulted in a significantly lower amount of the lighter protein fractions but unchanged amounts of the globulin fractions in atoxic nodular goitre.

Proteinuria in long term streptozotocin diabetes in rats

The total protein excretion rate of Munich-Wistar rats studied 80–90 days after the induction of diabetes with streptozotocin was more than double that of age-matched controls. The urine proteins consisted of only albumin and pre-albumin fractions. Dextran/inulin clearance ratios were not higher in the diabetic rats, and it is concluded that the proteinuria is not due to an increased glomerular permeability to protein. Kidney size was increased 1.7 fold and body weight was 60% of control. Glomerular filtration rate uncorrected for kidney or body weight was also not different from controls.

Polyacrylamide gel electrophoresis of cerebrospinal fluid proteins in children with nontumoral hydrocephalus.

The changes occurring in the electrophoretic pattern of the cerebrospinal fluid (CSF) proteins in 37 children with nontumoral hydrocephalus, and the protein pattern of 12 children without organic neurological disease, were studied with polyacrylamide gel electrophoresis. In nontumoral hydrocephalus protein was frequently increased in lumbar and ventricular fluid, in this condition, in intraventricular obstruction, the protein fractions showed a decrease in the prealbumin and an increase in the gamma-globulin fractions, and in extraventricular obstruction, the protein fractions showed a decrease in the prealbumin and an increase in the gamma-globulin and alpha 2-globulin fractions. With CSF proteins within normal ranges we found that in intraventricular obstruction, the proteins of the ventricular-CSF showed an increase in the prealbumin and/or albumin fractions, and in extraventricular obstruction, the proteins of the ventricular CSF showed an increase in the prealbumin and beta-globulin fractions, and the proteins of the lumbar CSF showed an increase in the prealbumin fraction. The diagnostic implications of these findings are discussed.

Prealbumin subfractions in brain tumors--with special reference to ontogenic changes (author's transl)

Prealbumin (PA) and its subfractions were analyzed to find their significance in the central nervous system from ontological and oncological points of view, using 7.5% polyacrylamide gel disc electrophoresis according to Ornstein51) & Davis.14) Tissues of 11 cases of normal human cerebrum, ranging from 5 or 6 months gestation to 64 years old, and 26 cases of brain tumor which included 12 gliomas, 8 meningiomas and 6 neurinomas were analyzed. Body fluids of 11 cases of brain tumor cysts, 12 ventricular cerebrospinal fluid with various neurological diseases, and 20 sera from the normal and the diseased were analyzed. 1. Tissue PA was subfractionated into five major peaks in the normal human brain, which we labeled PI through PV from the anodal side to the cathodal. In most of the brain tumors more than five peaks were subfractionated. 2. Concentration of PA subfractions changed in the developmental course in human brain. Among them, PIII and PIV changed remarkably in the fetal brain, namely, higher PIII and lower PIV levels were noted when compared to those of the adult brain. PI and PII showed no change during life. 3. Brain tumor had the same tendency as fetal brain regarding concentration of PIII and PIV subfractions. 4. PA III/IV ratios were compared as an index for showing some similarity in fetal brain and brain tumor tissues, and some tendency between them were observed. 5. PA fraction has been named as “neuronin” by Bowen et al.7)8)9)58) and as “SPR protein” by Kawakita32), and some of its subfractions have been identified, for instance, PI as S 100, PIV as 14-3-2 or antigen α, PV as a sensitive index for hypoxia or neurotubulin protein. PIII as well as PII still have not been identified. Further analysis of these proteins, immunochemically as well as electrophoretically, might answer the question of whether or not there are oncofetal proteins in the central nervous system.

The binding of thyroxine and tri-iodothyronine to plasma proteins in the chicken at the physiological pH

Amounts of 125I-labelled thyroxine (T4) and triiodothyronine (T3), representing approximately 50 per cent of the plasma concentration, were incubated with chicken plasma or serum, the proteins were separated by polyacrylamide gel electrophoresis at pH 7.4 and the distribution of the hormones determined. Seventy to 75 per cent of T4 migrated with albumin, about 10 per cent with the alpha-globulin and most of the remainder with a pre-albumin fraction. The percentage attached to the pre-albumin tended to increase with age. Neither barbiturate nor 8-anilino-1-naphthalene sulphonic acid (ANS) affected the distribution. About 50 per cent of T3 was associated with albumin, 30 to 40 per cent with the main alpha-globulin and most of the remainder with an adjacent globulin band. ANS reduced the percentage attached to albumin and increased that associated with the alpha-globulin. Barbiturate had no effect.

Thymic hormone-like restoration by human prealbumin of azathioprine sensitivity of spleen cells from thymectomized mice.

Gel filtration of fresh human serum revealed that over 95% of the thymic-hormone-like activity was present in the fraction representing the total albumin and prealbumin. Further studies demonstrated that the activity resided in the prealbumin fraction; albumin was inactive. Prealbumin was isolated from Cohn fraction IV-1 of pooled human plasma by extraction with aqueous buffer, fractionation with (NH4)2SO4, gel filtration on Sephadex G-150, and preparative disc gel electrophoresis. The final product was homogeneous by analytical gel electrophoresis was 40,000 times more active than the starting material in an azathioprine-sensitive rosette assay. Physical and chemical characterization studies showed that the isolated product was identical to authentic human plasma prealbumin. Isolation of the prealbumin fraction from sera of adult thymectomized mice revealed that the rosette activity was substantially lower than that isolated from sera of normal mice, suggesting a thymic dependence of the prealbumin activity. In vitro and in vivo bioassays of the fraction obtained prior to the final step of the purification procedure support the conclusion that prealbumin exhibits thymic hormone-like activity.

On the distribution of thyroglobin and larger iodoproteins in single rat thyroid follicles.

The distribution of the large iodoproteins in the lumen of thyroid follicles was tested. Samples of the colloid in rat thyroid follicles were obtained in vivo by micropuncture of superficial follicles in the isthmus of the gland. One sample was collected per follicle and several follicles were punctured in each gland. The protein composition of the samples was analysed by microgel electrophoresis. The protein separation patterns were recorded by microdensitometry after staining of the proteins and the amount of protein in the fractions was estimated from the densitometric recordings. Thyroglobulin (TG) and the aggregates of TG--here designated the S-TG fractions--were present in all samples of colloid. It was observed that there was a significant positive co-variation between TG and S-TG when the colloid contained only TG, S-TG and a prealbumin fraction. When the albumin-like protein was present in the samples the co-variation was less significant. The results show that the relative amount of the aggregates of TG is constant in the albumin free colloid and that these aggregates most probably are randomly distributed in the follicle lumen. The results further indicate that the aggregates of TG are non-randomly distributed in the larger peripheral follicles when the colloid contains the albumin-like protein.