Gel filtration and neutron activation analysis methods were combined in work aimed at the development of methods for the determination
of metals firmly bound to protein in human blood serum. The values obtained are semiquantative. A purely instrumental technique
afforded data on Cu, Fe, Zn, Al and Mn but values for the latter two only were determined since Cu, Fe and Zn are already
well known. The concentrations for Al and Mn were found to be 300 ppb and 0.4 ppb, respectively. Despite the high resolution
of Ge(Li) detectors, no other metals were found. For the determination of any others, radiochemical separations are necessary
and were used to obtain data on the following elements: As 3 ppb, Ag 3 ppb, Au 0.003 ppb, Cr 1 ppb, Sb 1 ppb, Co, Cd, Mo and
Sn could not be detected and upper limits were estimated to be 0.1 ppb, 2 ppb, 2 ppb, and 1 ppb, respectively.
A radioimmunoanalytical (RIA) method was elaborated for the determination of ferritin in human blood serum in clinical practice. Placental ferritin separated from the human after-delivery placental and antibodies against the human placental ferritin obtained by the immunization of rabbits with this antigen were used. The whole complex of basic conditions and parameters of the RIA method was tested including the estimation of the region of normal values and clinical tests. The method elaborated was compared with the commercial kit Ferritin RIA Amersham, code IM 1051, chosen as reference kit. The results of the determination of control parameters as well as ferritin levels obtained by the method elaborated exert good agreement with the reference kit and correspond to requirements for routine RIA practice.
Differential scanning calorimetry (DSC) has been employed to study the thermal denaturation processes of the main protein
fractions of blood serum. These processes have been compared for albumins (nondefatted (HSA) and fatty acid free (HSAf)),
α,β-globulins, γ-globulins, and their mixtures in aqueous (pH 6.5) and buffer (pH 7.2) solutions. The results have indicated
that α,β-globulins inhibit γ-globulins’ aggregation in buffer solutions. The decrease of stability of HSA and HSAf aqueous
solutions has been observed in the presence of γ-globulins. The mixtures of albumins and γ-globulins have revealed the tendency
to ready aggregation in water. Moreover, the results have suggested that neither γ-globulins nor albumins severely change
the stability of α,β-globulins.
Authors:Viera Revajová, D. Magic, M. Levkut, Ĺ. Bindas, M. Horváth, R. Kašteĺ, J. Pistl, and J. Šajbidor
Oral administration of n-3 polyunsaturated fatty acids (PUFA) to piglets slightly enhances the immune response. As compared to the control, in the experimental piglets the absolute values of monocytes in the peripheral blood were significantly increased (P < 0.05), while the metabolic activity of phagocytes and the number of lymphocytes within the individual subpopulations were slightly higher. The level of growth factors, determined on the basis of somatomedin in the blood serum, was significantly higher in the experimental group (P < 0.05). n-3 PUFA interfere with the synthesis of prostaglandins and influence the metabolism of fatty acids. This finding may contribute to the therapy of inflammatory processes influencing immune and growth factors in piglets.
Authors:Márta Erdélyi, M. Mézes, and Györgyi Virág
Glutathione peroxidase enzyme superfamily plays significant role in the elimination of reactive oxygen free radicals in the animals. Many characteristics of these proteins have been revealed already, but their regulation is still not known. Several data suggest that some environmental factors have certain regulatory effect, while others propose strict genetic regulation.In this report we present four different environmental induction models in which New Zealand white rabbits were used as experimental animals. In three models, free radical load of different origin, lipidperoxide load, application of a glutathione depletor or a prooxidant agent, was introduced. Beside these negative models a positive model was also constructed in which additive selenium was supplied.Glutathione peroxidase activity was measured in blood serum, erythrocyte haemolysate and liver. Reduced glutathione, and malondialdehyde concentration in the liver were also determined.According to the results, the established models are capable for analysing the enzyme activity´environmental interactions.
Authors:Stefania Iwańska, Danuta Strusińska, and W. Zalewski
When evaluating the effects of yeast culture (Saccharomyces cerevisiae1026) supplied with or without a vitamin premix and mineral bioplexes on some intermediates and end-products involved in the synthesis of milk constituents in 30 early-lactation Black and White Lowland cows, no significant differences were found in the glucose level, mineral contents and enzyme activities of the blood serum. The effect of yeast culture on the availability of minerals for milk synthesis depended upon the dynamics of degradation of mineral bioplexes in the rumen and the cows' mineral status. The insignificant increase found in blood total protein content and the simultaneous small differences in blood urea nitrogen (BUN) and milk urea nitrogen (MUN) values in cows supplied with the yeast culture were probably associated with a high ammonia incorporation into microbial protein in the rumen, which increased protein supply for milk protein synthesis and decreased the nitrogen loss.
Authors:S. Fekete, Tibor Gaál, M. Hegedűs, F. Janza, and L. Zöldág
The nutritive value of dog foods declared by the manufacturer as nutritionally complete and balanced can be best assessed by feeding trials with dogs. A protocol of a feeding trial has been developed and tested with working dogs fed two different commercial complete and balanced diets for 8 weeks. The parameters used for evaluating the effect of diets were general health status, body and hair coat condition, change of body weight, haematological parameters (white blood cell (WBC) count, red blood cell (RBC) count, haemoglobin, packed cell volume), and biochemical parameters in blood serum (alanine aminotransferase, urea, albumin). The trial protocol proved to be appropriate to monitor the dogs' nutritional status and to reveal differences between diets. This method of evaluation is recommended for use in supporting the nutritional claims (labelling) of dog foods.
Authors:J. Juskiewicz, M. Wróblewska, K. Zhaki, Z. Zdunczyk, and L. Hussein
The objective of the experiment was to determine whether small amounts of proanthocyanidins (0.1 and 0.3%) may increase the antioxidative properties of the rat diet without exerting an antinutritional effect. Proanthocyanidins of faba bean seed coats were extracted with a mixture of acetone and water (70:30) and lyophilized. The amount of proanthocyanidins was two- or fourfold higher in the experimental diets as compared to the control diet. The addition of proanthocyanidin extract had no significant effect on the coefficients of digestibility of crude protein, daily nitrogen retention and the coefficient of biological value of diet protein. In the blood serum of rats fed diets supplemented with proanthocyanidin extract, there was a slightly higher content of vitamin E and alanine aminotransferase activity, while the content of vitamin A and aspartate aminotransferase activity were similar to those of the control group. In the contents of the rat gut (caecum), a lower activity of â -glucuronidase was found as compared to the control group, whereas â -galactosidase was unaffected. The addition of proanthocyanidin extract to diet caused a decrease in the malondialdehyde content in the heart, kidneys, erythrocytes and blood plasma of rats. The results obtained indicate that the amount of proanthocyanidins used did not exert any antinutritional effects, but extended the pool of diet antioxidants and beneficially affected the activity of the large bowel microflora.
Authors:Małgorzata Dżugan, Marcin Lis, Maria Droba, and Jerzy Niedziółka
The aim of the study was to determine the toxicity of cadmium ions in chick embryos, using plasma hydrolytic enzyme as its biomarker. Hatching eggs (n = 300) from Ross 308 broilers were incubated under standard conditions. On day 4 of incubation, 50 μl of saline solution, containing Cd ions at a concentration from 0 (control group) to 24 μg, was injected in ovo into the egg albumen. The results indicate that the administration of cadmium at doses exceeding 1 μg/egg caused a gradual decrease in hatchability, with an LD50 of 3.9 μg/egg. The greatest differences between the groups in the enzymatic activities studied were found for N-acetyl-β-D-glucosaminidase (NAG), β-D-mannosidase (β-MAN) and arylsulphatase (ARYL). Compared to the control group, in the blood serum of chicks from the groups receiving 3, 6 and 12 μg Cd/egg the NAG activity increased by 79, 108 and 54% and β-MAN activity by 33, 119 and 108%, respectively. Exposure to cadmium at a dose of 1 to 6 μg per egg caused an about 60% increase in ARYL activity while a dose of 12 μg decreased the activity by about 35% below the level observed in the control group. These findings show that cadmium has a similar toxicity mechanism in mammals and birds, which opens the possibility of using NAG activity as a biomarker of the cytotoxic effect of cadmium in birds.
Authors:A. Al-Ammar, E. Reitznerová, and R. Barnes
Two sample treatment methods are evaluated to provide accurate boron determination at low concentrations in biological and botanical samples. The first approach is a hot 1M nitric acid extraction of boron from the sample. The second technique uses wet digestion with concentrated sulfuric acid. The accuracy of the procedures was demonstrated with botanical and animal reference materials (Corn Bran RM 8433 and Whole Egg Powder RM 8415). At least three results are given for each reference material. Two are direct measurements of B using independent 10B and 11B calibration curves with a Be internal reference, and the third is obtained by isotope dilution mass spectrometry (IDMS). The 10B and 11B values are consistent for both acid treatment procedures. The IDMS results also are consistent. Overall results for Whole Egg Powder and Corn Bran RM's match the best-estimate values within their confidence intervals. These results demonstrate the ability to measure B accurately at the 0.3 µg/g concentration range. Thus, low-level B samples can be analyzed with accuracy and precision by the two approaches. These methods introduce very little dissolved organic carbon (DOC) in the final solution and allow the use of large (2 g) sample aliquots. Direct introduction of biological fluids including whole blood serum also was evaluated critically for the determination of B.