Immunostaining of human spermatozoa with tubulin domain-specific monoclonal antibodies. Recognition of a unique beta-tubulin epitope in the sperm head

Summary Four monoclonal antibodies that discriminate between structural domains of alpha-(TU-01, TU-04) or beta-(TU-06, TU-12) tubulin and a polyclonal anti-tubulin antibody were used for immunostaining of human spermatozoa using immunofluorescence microscopy. Specificity of antibodies was confirmed by immunoblotting experiments. Antibodies TU-01 and TU-06 uniformly stained the whole tail and the neck, whereas antibodies TU-04, TU-12 showed differential distribution of corresponding epitopes in the stable arrays of flagellar microtubules. Of the monoclonal antibodies used, only TU-12 against the antigenic determinant on C-terminal domain of -tubulin showed strong reactivity with the equatorial segment of the head. The results document a differential exposure of tubulin epitopes at the single-cell level and suggest the existence of distinct tubulin populations in various structural compartments of the human spermatozoon.     

Kiwifruit micropropagation through callus shoot-bud induction

Summary A profitable system for the establishment of morphogenic callus cultures and indirect shoot induction and development was accomplished from nodal shoot segments obtained from adult and micropropagated plants of kiwifruit (Actinidia deliciosa [Chev.] Liang and Ferguson, var.deliciosa) cv. “Hayward”. The effects of medium composition, cytokinin levels, dilution of salts, and type of callus derived from the cultured primary explants were studied. Medium composition as well as type of callus greatly affected organogenic responses.     

Oenanthe aquatica (L.)Poir.: Seed reproduction,population structure,habitat conditions and distribution in Czechoslovakia

The paper summarizes data concerning the biology and ecology ofOenanthe aquatica. The species is commonly distributed all over Czechoslovakia, from lowlands to the submontane belt, especially in fishpond and river basins.O. aquatica shows no special relationship to the chemical and physical soil properties, and is well adapted to habitats with a changing water level. The reproduction ofO. aquatica depends on emerging of the bottom. The most developed populations are formed by biennial plants and arise in the year following summer or autumn drainage.     

A contribution to optimal control of fed-batch biochemical processes

The problem of looking for high efficient modern control strategies in fermentation technology is very urgent, nowdays. Particular attention should be paid to the processes in fed-batch mode. Both, optimal feedforward and feedback control approaches are suggested. A contribution is considered to have been made in the feedback control where continuous and discrete versions are treated as well. The control laws are carried out by a variation calculus problem and a polynomial pole placement synthesis solution, respectively. All the algorithms result in an optimal substrate feed rate profile. On the basis of recursive least squares identification of the model coefficients an adaptive discrete-time control strategy is proposed. Some satisfying simulation results are dealt with.     

Agonist binding to purified glycine receptor reconstituted into giant liposomes elicits two types of chloride currents.

Using 'inside-out' membrane patches obtained from reconstituted giant liposomes containing purified glycine receptor from rat spinal cord, we have detected chloride currents elicited in response to the presence of the agonists glycine or beta-alanine. Regardless of the agonist employed, two different patterns of single channel currents could be detected, which differ in their main conductance, complexity of substates and opening frequency. In agreement with the expectations of glycine receptor heterogeneity suggested recently at the mRNA and cDNA level, our results indicate the existence of functionally different glycine receptors in the adult rat spinal cord.     

Proteins under extreme physical conditions

Life on earth is ubiquitous within the limits from -5 to 110 degrees C for temperature, 0.1 to 120 MPa for hydrostatic pressure, 1.0 to 0.6 for water activity and pH 1 to 12. In general, mutative adaptation of proteins to changing environmental conditions tends to maintain 'corresponding states' regarding overall topology, flexibility and hydration. Due to the minute changes in the free energy of stabilization responsible for enhanced stability, nature provides a wide variety of different adaptative strategies. In the case of thermophilic proteins, improved packing densities are crucial. In halophilic proteins, decreased hydrophobicity and clustered surface charges serve to increase water and salt binding required for solubilization at high salt concentration. In the case of barophiles, high-pressure adaptation is expected to be less important than adaptation to low temperatures governing the deep sea. Nothing is known with respect to the mechanisms underlying psychrophilic and acidophilic/alkalophilic adaptation.     

Members of the alpha-amylase inhibitors family from wheat endosperm are major allergens associated with baker's asthma

We have identified the major antigens or IgE binding components from wheat flour. Thirty-five sera from patients with baker's asthma were used to analyze the reaction with wheat salt-soluble proteins. We found a 15 kDa SDS-PAGE band which reacted with all sera tested. Purified members of the alpha-amylase inhibitor family, which are the main components of the 15 kDa band, were recognized by specific IgE when tested with a pool of reactive sera. Immunodetection after two-dimensional electrophoretic fractionation of crude inhibitor preparations from wheat endosperms also detected several inhibitor subunits as major low-molecular-weight allergens.     

Distribution of cells in different stages of the cell cycle and some age-related physiological characteristics in dependence on the dilution rate in chemostat cultures ofCandida utilis

Position of cells in their cell cycle was determined microscopically in chemostat cultures ofCandida utilis. Proportion of cells in phase G₁ decreased in a linear manner from 86% to 58% with dilution rate. Proportion of cells in phase S increased in the same range ofD from 5.6 to 13.5% and in the (G₂+M) phase from 8.4 to 28.5%, again linearly. Differential centrifugation was used to separate chemostat cultures to mother and daughter cells. Analyses showed that, relative to mother cells, daughter cells contain 2.1–11.9% more protein and 25.5–34.6% more RNA in dry matter. Their mass is 34.4–5.6% lower and volume is 154–19% smaller.     

The effect of soluble glucan derivates on spleen colony-forming units in sublethally irradiated mice

We examined the effects of 5 soluble derivatives of yeast glucan on the formation of exogenous (CFU-S) and endogenous (E-CFU) colony-forming units in the spleens of sublethally irradiated (⁶⁰Co, 6.5–7.0 Gy) mice of two inbred strains. For the estimation of CFU-S, glucans were administered intravenously either to donors or recipients of spleen cells 24 h prior to irradiation or removal of the spleen. The number of CFU-S was increased when both the donors and recipients were treated with glucan; the highest increase was obtained with glucans S, P and K. All glucan preparations increased significantly also the number of E-CFU even when administered 90 min after irradiation. There exist differences in the response to the stimulatory effect of glucans among individual mouse strains. Thus, for example, the stimulatory effect of glucan KM on the E-CFU number was significantly more pronounced in strain A/Ph than in strain C57B1/6.     

Genetic stability of the antagonistic character ofEnterococcus faecalis ssp.liquefaciens and the detection of a new inhibitory bacteriocin-like substance

The inhibitory capacity of strain S-48 ofEnterococcus faecalis ssp.liquefaciens was studied. The strain produces a broad-spectrum peptide antibiotic (AS-48) that has been characterized elsewhere. The isolation of mutants from S-48 after mutagenic treatment revealed another inhibitory substance which remained masked in the wild strain. The protein nature and restricted spectrum of this substance points to its being a bacteriocin (Bc-48).