Effect of periodate oxidation on properties of antibodies

The subject of the present research is the investigation of the influence of sodium periodate on the properties of immunoglobulin G molecules. It is shown that 100 and 300 M of periodate cause a slight enhancement of the sedimentation coefficient which is a result of the higher protein density. However high concentrations (2000 M) of periodate decrease sedimentation coefficient considerably and disturb the protein structure homogeneity. Studies of the immunologic activity in the periodate-treated antibodies by the reaction of passive hemagglutination showed that in low concentrations it did not decrease significantly the activity but with an increase in the concentration up to 2000 M the activity lowered. The conjugation of antibodies with the enzyme markers was fulfilled due to periodate oxidation. The conjugates obtained were successfully used for improving sensitivity of the precipitation line in immunologic tests.     

Effect of dye quality and the method of preparing a Schiff reagent on the staining intensity and DNA-fuchsin absorption spectra in performing the Feulgen reaction

Dependence of the staining and absorption spectra of DNA-fuchsin on the dye quality and modes of preparation of the Schiff-reagent was studied. It is shown that many dyes supplied by different producers do not suit the purposes of quantitative cytochemical investigations. Before use the absorption spectra of the DNA-fuchsin must be taken using the standard object. The use of dyes possessing a multi-peak absorption curve should be avoided.     

Effect of periodate oxidation on the structure and properties of glucose oxidase.

In order to elucidate the molecular structure of glucose oxidase (beta-D-glucose: oxygen 1-oxidoreductase, EC 1.1.3.4) and the roles of its carbohydrate moiety, chemical, physiochemical and immunological experiments were performed with enzyme samples before and after periodate oxidation. Hydrodynamic parameters indicated that the native enzyme was a globular protein with values of 1.21 for the frictional ratio and 43 A for the Stokes radius. The enzyme contained about 12% carbohydrate by weight, of which the main component was mannose. The periodate treatment decreased the carbohydrate content to about 40% of its original value. Slight modifications were detected in the absorbance spectrum and the content of arginyl residue. However, no significant alteration was brought about by this treatment in the catalytic parameters, immunological reactivities of the gross structure, not in the secondary and quaternary structures of the protein moity. Thermal denaturation temperature (about 72.5 degrees C) and the enthalpy of denaturation (about 450 kcal/mol) were common to the native and the periodate-oxodozed enzymes. The native was found to be quite resistant to sodium dodecyl sulfate and fairly stable to urea and heating. The periodate-oxidized enzyme was also stable to heat treatment, but it showed a diminished stability when denaturing agents were present. Kinetic analyses of the thermal inactivation processes showed that the entropy of activation was greatly decreased by the denaturing agents, especially in the case of the periodate-oxidized enzyme. It is concluded that the carbohydrate moiety of the enzyme plays a role in increasing the stability of the protein moiety, but does not directly participate in the catalytic activity, the immunological reactivity, or in maintaining the conformation of the enzyme protein.     

Effect of periodate oxidation on the polysaccharide content and microaggregate stability of rhizosphere and non-rhizosphere soils

Summary The relationship between the stability of soil microaggregates in water and the polysaccharide content was examined in rhizosphere and non-rhizosphere samples from a pot experiment using three soils that had grown peas, barley or grasses. The polysaccharide was oxidised and removed using 168h treatment with 0.02M periodate followed by 6h with 0.1M tetraborate. The decrease in polysaccharide content, measured as change in residual reducing sugars, was compared with the stability of soil microaggregates (ca 45m) in water, determined by a turbidimetric method.Total C, N and polysaccharide contents of rhizosphere soils were greater than those for the bulk soil, but the water stability of aggregates was not increased compared to unplanted controls. Periodate oxidation removed a large proportion (59–95%) of the polysaccharide and increased aggregated disruption, but there was no clear relationship between the two measurements. In rhizosphere soil, polysaccharides appreared to make less contribution to aggregate stability than polysaccharide in the bulk soil. The relatively small effect of rhizosphere polysaccharides is probably related to their presence as comparatively massive plant remains and debris; this contrasts with the decomposed and transformed material in the bulk soil.     

The periodate oxidation of amino acids with reference to studies on glycoproteins

1. All α-amino acids are oxidized by periodate, but at different rates. 2. The rates of oxidation of individual α-amino acids vary with pH. In general, oxidation proceeds more rapidly at alkaline pH. 3. Serine, threonine, cysteine, cystine, methionine, proline, hydroxyproline, tryptophan, tyrosine and histidine are rapidly and extensively oxidized by periodate. 4. Cysteine, cystine, methionine, tryptophan, tyrosine and histidine are oxidized by periodate when they are substituted in the carboxyl and amino groups, as in a polypeptide chain.     

Effect of exercise intensity on 24-h energy expenditure and nutrient oxidation.

The aim of this study was to determine the effects of exercise at different intensities on 24-h energy expenditure (EE) and substrate oxidation. Sixteen adults (8 men and 8 women) were studied on three occasions [sedentary day (Con), a low-intensity exercise day (LI; 400 kcal at 40% of maximal oxygen consumption) and a high-intensity exercise day (HI; 400 kcal at 70% of maximal oxygen consumption)] by using whole room indirect calorimetry. Both 24-h EE and carbohydrate oxidation were significantly elevated on the exercise days (Con < LI = HI), but 24-h fat oxidation was not different across conditions. Muscle enzymatic profile was not consistently related to 24-h fat or carbohydrate oxidation. With further analysis, it was found that, compared with men, women sustained slightly higher rates of 24-h fat oxidation (mg x kg FFM(-1) x min(-1)) and had a muscle enzymatic profile favoring fat oxidation. It is concluded that exercise intensity has no effect on 24-h EE or nutrient oxidation. Additionally, it appears that women may sustain slightly greater 24-h fat oxidation rates during waking and active periods of the day.     

Effect of periodate oxidation on the detection of antibodies against the M-antigen of histoplasmin by enzyme immunoassay (EIA) inhibition

An enzyme immunoassay (EIA) inhibition method was devised that detects antibodies against the M-factor, a major protein antigen in histoplasmin. Purified M-factor still contains a large amount of a carbohydrate antigen impurity, but this can be inactivated by periodate oxidation. The resulting product improves the performance of the EIA to detect anti-M by (a) giving a more specific measurement, (b) increasing the percent inhibition in sera that contain antibodies to M-factor, and (c) increasing the precision in measuring negative reactions. An inhibition of 20% or greater is recommended for recording a result as positive.     

Effect of duration and intensity of infection with Echinostoma audyi on survival of Lymnaea rubiginosa exposed to copper sulfate.

Lymnaea rubiginosa infected with 1 to 5 miracidia of Echinostoma audyi for 1, 2, 3, and 5 weeks, and snails infected with up to 10 or 20 miracidia for 3 weeks, were exposed to copper (Cu) as CuSO4 2 hr and allowed to recover 48 hr. In general, snails ininfected with up to 5 miracidia of E. audyi were no more susceptible to Cu than noninfected snails. Snails infected with up to 10 miracidia for 3 weeks were 1.3 times more susceptible to Cu than noninfected snails. However, snails infected with up to 20 miracidia for 3 weeks were not more susceptible to Cu, possibly because infection with such a large number of parasites eliminated the most susceptible snails from the population prior to exposure to Cu.