Uptake of microperoxidase by segmenting rat ova in vitro

Summary Uptake and incidence of microperoxidase (Sigma) in the rat ova during cleavage at the 1-, 2-, 8-cell and blastocyst stages were studied after 30 min incubation with the enzyme (molecular weight of 1,900 and size of molecule of 2 nm).Evidence was furnished of the presence of a small amount of reaction product of microperoxidase in the zona pellucida and its local concentration in some parts of the perivitelline space. The larger part of surface of the ovum, however, was free of microperoxidase. In the cytoplasm microperoxidase was found in pinocytotic vesicles, less frequently in large vacuoles and starting with the eight-cell stage in secondary lysosomes. The largest amount of microperoxidase was ascertained at the stage of blastocyst, chiefly in the cells of the trophoblast. In the cells of the embryoblast, on the contrary, microperoxidase was found but occasionally. The reaction product of microperoxidase was also present in the intercellular space of ova in negligible amount, and likewise on the side of blastocysts' cavity. In comparison with the ingestion of horseradish peroxidase the incidence of microperoxidase in the segmenting rat ova was less frequent.     

Origin of sclerotia-like cells in submerged Claviceps purpurea producing clavine alkaloids

Ultrathin sectioning of submerged mycelium of Claviceps purpurea Tul. producing clavine alkaloids revealed yeast-like budding resulting in asexual sporesblastospores. These deciduous spores were born by extended hyphal cells and retained the same ultrastructure of cell organelles. Both the extended hyphae and the blastospores resembled the cells of ergot sclerotial tissue. A surface culture of C. purpurea Tul. producing no alkaloids was used as a reference.     

The effect of splenectomy, produced chemically by means of ethylpalmitate, on the formation of heteroagglutinins in the rat]

The formation of heteroagglutinins against human O-erythrocytes was pursued in Wistar rats splenectomized "chemically" by means of intravenous injections of ethyl palmitite. Contrary to some data in literature the initial inhibition in the formation of antibodies was replaced by a significant titre increase ranging from the 8th to the 14th day following immunization. The tween 20 contained in the emulsion of ethyl palmitic did not influence the formation of heteroagglutinins. The examinations show that a possible use of palmitic ethyl for immunosuppression is questionable.     

The occurrence of beet mild yellowing virus in Czechoslovakia

The occurrence of beet mild yellowing virus (BMYV) on feeding- and sugar-beet in Czechoslovakia has been proved. The virus was transmitted by aphidMyzus persicae (Sulz.) on indicator plantsSinapis alba L.,Capsella bursa-pastoris Medik, andClaytonia perfoliata Donn and from these plants back to sugar-beet cv. ‘Dobrovická A.’ A weed plantRaphanus raphanistrum L. was identified as a new natural host plant of BMYV. The virus was identified in ten of twelve biologically examined samples of beet with BMYV-like symptoms, which were collected at various places in Czechoslovakia     

Gylcerol-3-phosphate shuttle and its function in intermediary metabolism of hamster brown-adipose tissue.

1. Brown adipose tissue of the hamster possesses high specific activities of soluble, cytoplasmic NAD-linked, as well as mitochondrial flavin-coupled, glycerol-3-phosphate dehydrogenases. The ratio of the two enzyme activities is high (close to 1), when compared with other tissues of the hamster. 2. In the presence of rotenone, NADH is oxidised very poorly by homogenates of brown adipose tissue. A high rate of oxidation is obtained upon further addition of dihydroxyacetone phosphate, which itself is negligible oxidised. When followed fluorimetrically glycerol 3-phosphate can also be observed to induce NADH oxidation, but only after a significant lag time. Similar results are obtained with isolated mitochondria plus high-speed supernatant. With high-speed supernatant alone, only dihydroxyacetone phosphate has any effect, whereas with isolated mitochondria neither dihydroxyacetone phosphate nor glycerol 3-phosphate induce any NADH disappearance. 3. Respiration induced by NADH plus dihydroxyacetone phosphate in homogenates equals 56% of the respiration induced by glycerol 3-phosphate alone. 4. Respiration induced by NADH plus dihydroxyacetone phosphate, as well as that induced by glycerol 3-phosphate, is inhibited by the same concentrations of inhibitors as are required for inhibition of the mitochondrial dehydrogenase i.e. EDTA, long-chain unsaturated fatty acids, long-chain fatty acyl CoA esters. 5. In isolated brown adipocytes in the presence of rotenone, norepinephrine significantly inhibits respiration induced by glycerol 3-phosphate. 6. The results obtained are discussed with respect to the role of glycerol 3-phosphate as an electron sink for cytosolic reducing equivalents to maintain a low level of extramitochondrial NADH. A means of maintaining a level of glycerol 3-phosphate adequate for triglyceride synthesis is also considered.     

Proteolytic activity associated with human erythrocyte membranes. Self-digestion of isolated human erythrocyte membranes.

At least two kinds of enzymes are active in the proteolytic self-digestion of erythrocyte membranes. The specific activities of these enzymes do not decrease with repeated washings of purified stroma. The effects of a variety of inhibitors on the membrane preparation's capacity to digest 125-I-labelled casein, covalently linked to latex beads, have been examined. Pepstatin-inhibitable enzyme, active at low pH, digests the membrane extensively to small polypeptide fragments. Spectrin, located at the internal part of the membrane, is readily degraded. Diisopropylfluorophosphate-inhibitable enzyme, active at pH 8-9, has only limited digestive capacity. Some of the membrane components, such as the small molecular weight glycoproteins, are resistant to digestion. The restricted capacity of digestion is due to the membrane molecular arrangement; increased disaggregation removes the restriction and increases the activity. Spectrin is not digested unless the membrane topography is disrupted by NP-40 neutral detergent. These observations suggest that the enzymes active at basic pH are located external to the cell. Intact cells do possess a limited capacity to degrade 125-I-labelled casein when their surfaces are brought into contact with substrate-coated beads.     

Penicillinamidohydrolase inEscherichia coli

Substrate specificity of the bacterial penicillinamidohydrolase (penicillinacylase, EC 3.5.1.11) fromEscherichia coli was determined by measuring initial rates of enzyme hydrolysis of different substrates within zero order kinetics. SomeN-phenylacetyl derivatives of amino acids and amides of phenylacetic acid and phenoxyacetic acid of different substituted amides of these acids or amides, structurally and chemically similar to these compounds, served as substrates. Significant differences in ratios of initial Tates of the enzyme hydrolysis of different substrates were found when using a toluenized suspension of bacterial cells or a crude enzyme preparation, in spite of the fact that the enzyme is localized between the cell wall and cytoplasmic membrane, in the so-called periplasmic space.N-phenylacetyl derivatives are the most rapidly hydrolyzed substrates. Beta-phenylpropionamide and 4-phenylbutyramide were not utilized as substrates. The substrate specificity of the enzyme is discussed with respect to a possible use of certain colourless compounds as substrates, hydrolysis of which yields chromophor products suitable for a simple and rapid assay of the enzyme activity.     

Accumulation of Natural Radionuclides in the Bottom Sediments and by Aquatic Organisms of Streams

The accumulation of natural radioisotopes by aquatic organisms and bottom sediments was studied in two small rivers, one uncontaminated and the other polluted by effluents of uranium ore mining and milling. Parabolic regressions between the water and uppermost sediment content of both uranium and total beta activity (corrected for ⁴⁰K content) is presented and demonstrates the water cumulative capacity of organically rich sediments. For ²²⁶Ra no significant differences between sediments with different organic content were found. The ²²⁶Ra content of bottom sediments is expressed as a power function of radium and calcium concentration in the water. In the given reaches, 53% and 85% of uranium and over 90% of ²²⁶Ra and total beta activity (corrected for ⁴⁰K) was accumulated in the upper two centimetres of sediments and biomass of aquatic vegetation. Filamentous algae, plankton, aquatic bryophyta and macrophyta from the present and other published data showed rather higher cumulation capacities as compared with bottom sediments.