Study of properties of structural subunits of IgM immunoglobulin obtained by reduction with 2-mercaptoethanol or by oxidative sulphitolysis

IgM was isolated from pig serum by isoelectric precipitation and gel filtration. Different methods of breaking down the disulphide bonds and of isolating subunits of the IgM molecule—oxidative sulphitolysis and reduction by 0.1m 2-mercaptoethanol in the absence of a disaggregating agent, oxidative sulphitolysis in the presence of 6m urea and reduction by 0.3m 2-mercaptoethanol in medium containing 6m and 8m urea—were compared. Degraded material was separated by gel filtration on Sephadex G-100 or G-200 in 0.05m formic acid with 6m or 8m urea. Oxidative sulphitolysis or reduction by 0.1m 2-mercaptoethanol without a disaggregating agent did not yield pure H andl chains. Oxidative sulphitolysis was the more effective. Oxidative sulphitolysis in 6m urea medium severely damaged the material. Reduction of IgM by 0.3m 2-mercaptoethanol in 6m or 8m urea also altered its immunochemical properties. The possible presence of light chains in the heavy chain fraction cannot likewise be excluded in this case. The results are in agreement with experiments showing that the molecular weight of the IgM heavy chain is greater than that of the IgG heavy chain.     

Study of the effect of volatile substances from cereal roots

We traced the liberation and biological effect of volatile substances released from the roots of cereals,i. e. barley, wheat, rye and oats, on seedlings of the same and other plant species. Experiments were carried out in a closed glass apparatus with a static or circulating atmosphere in which the CO₂ and O₂ were permanently absorbed and supplemented, respectively. In some experiments the air was bubbled through water or through solutions of boric acid, barium hydroxide and potassium permanganate. The roots of all four cereals tested released volatile substances with a biological activity which appeared to be non-specific with respect to plant species. The effect of volatile substances was partially decreased by bubbling through water, barium hydroxide and boric acid and was completely removed after passing through the solution of potassium permanganate. Volatile substances liberated from roots of barley inhibited elongation of roots and coleoptile, decreased SH-group content and caused excessive formation of root hairs as well as inhibition of both dry matter production and respiration of roots of rye seedlings. Ethylene was found in the atmosphere of experimental vessels.     

Some properties of pea enation mosaic virus isolated from field pea and broad bean plants in Bohemia

Pea enation mosaic virus (PEMV) was isolated from disea sed field pea (Pisum sativum L.ssp. arvense A.Gr.) and broad bean (Faba vulgaris Moench) plants grown as filed crops at Bohumilice in Bohemia. The virus proved to be pathogenic for the following plant species:Pisum sativum L. cv. Raman,Faba vulgaris Moench,Lens culinaris Med.,Vicia sativa L.,Lathyrus odoratus L.,Glycine soja L.,Phaseolus vulgaris L.,Chenopodium amaranticolor Coste andReyn,Nicotiana clevelandi Gray,Trifolium incarnatum L. The dilution end point of the isolate was higher in pea plants (10^?4) than in broad bean plants (10^?2). The thermal inactivation point was 65–68° and the longevityin vitro between 10 and 14 days. According to the host range, symptoms on pea plants and physical properties the virus isolate studied resembles some isolates described in the U.S.A. and represents a PEMV strain different from those reported so far in Czechoslovakia.     

Underground dry weight in the grassland communities ofArrhenatheretum elatioris alopecuretosum pratensis R. Tx. 1937 andMesobrometum erecti stipetosum Vicherek 1960

The meadow plant communities,Arrhenatheretum elatioris alopecuretosum pratensis R. Tx. 1937 andMesobrometum erecti stipetosum Vicherek 1960, were chosen for investigations of the underground plant parts. Apparent differences in underground dry weight and its seasonal changes in both the communities were observed. Differences in the soil environment in different periods of the year are reflected in the character of time changes in underground dry weight. The soil environment affects not only the total underground biomass and their changes in time, but also the activity of soil microflora and, consequently, the decomposition rate of dead underground plant parts.     

Antibacterially active substances

Synthesis of two hydroxy-derivatives of nalidixic acid as a result of microbial transformation was demonstrated in certain species of the genusAspergillus. Aspergillus alliaceus produced 7-hydroxy-nalidixic acid andAspergillus niger 6-hydroxy-nalidixic acid. It was demonstrated that the antibacterial activity of both hydroxy-derivatives (tested inEscherichia coli) was lower than that of the initial nalidixis acid.     

Electrophoretic investigation of proteins in different root zones ofvicia faba L.

The differentiation of tissues is closely connected with the proteosynthesis. One can therfore assume that tissues with different types of cell growth (meristematic or elongation growth) and with different degrees of differentiation are different in their protein composition. In order to compare the protein composition of different plant organs, the method of disc electrophoresis on a polyacrylamide gel has been used by some authors. As compared with other methods used up to now, e. g. isolating proteins on DEAE cellulose or in Sephadex, this method does not need so much material and its resolution ability seems to be higher. It is also quicker and enables the study of several samples simultaneously. Its disadvantage is that proteins can be identified mainly by means of Rf and their quantity, measured from the intensity of staining of individual fractions in the gel, which may be misleading due to different sorption capacity of different proteins (Fri?Fri?ová 1967). None the less, it is good for comparison of protein composition of individual parts of the plant body. Different methods have been used to compare protein composition of individual growth zones in roots.Barsky,Ivanov andPushakova (1965) used luminiscence microscopy and found that in maize roots it is not possible to find substantial differences by this method.Morgan andReith (1954) arrived at similar conclusions. On the other hand,Steward et al. (1965) andMorris (1966) found qualitative differences in protein composition of different parts of pea roots using acrylamide electrophoresis. The results of the last named authors show considerable discrepancies in details, due perhaps to a different method of extraction (buffer, pH, purifying method). We have used acrylamide gel electrophoresis for investigating proteins in precisely defined growth zones of theVicia faba root.     

Use of Carbowax as a high molecular weight nonprotein substance for cultivation of lymphoid cell and study of their morphology

Lymphoid cells prepared from rabbit popliteal lymph nodes were cultivated in medium in which the high molecular weight component was replaced by Carbowax 20 M. This substance, in 0.2% concentration, proved to be a substitute for the high molecular weight component in short-term cultivation, and the viability results were similar to those in medium containing normal rabbit serum or 0.5% human serum albumin. In addition, mitoses and the formation of large basophilic cells resembling the blastoid cells induced during cultivation with phytohaemagglutinin were observed in this medium. The results are discussed.     

Secondary responsein vitro to antigen ΦX 174 phage

In a model secondary reactionin vitro, a correlation was demonstrated between the size of the antigen dose used for the prestimulation of spleen tissue donors and the type of antibodies formed in the anamnestic reaction. After a small dose of antigen ΦX 174, the antibody response three days after prestimulation was of the 19 S (IgM) type, but later secondary contactin vitro (after four months) did not produce a 19 S anamnestic reaction. After large primary doses of antigen, a short interval between primary and secondary contact led to the formation of 19 and 7 S type antibodies, while after a long interval only 7 S (IgG) type antibodies were formed. The results are discussed in relation to differences in the size of the antigen dose needed to induce short-term 19 S and long-term 7 S immunological memory.