Histochemical demonstration of the intestinal hetero-β-galactosidase (glucosidase)

Summary The histochemical demonstration of hetero--galactosidase (glucosidase) has been attempted in sections and zymograms of rabbit, monkey and human intestine and of rat kidney.The leakage of this enzyme from unfixed sections was prevented by the use of cold microtome sections adherent to semipermeable membranes. Methods with -D-glucosides and galactosides of 6-Br-2-naphthol (postincubation azocoupling with Fast Blue B as well as simultaneous azocoupling with hexazonium-p-rosaniline), of -naphthol (simultaneous azocoupling with hexazonium-p-rosaniline) and of 4-Cl-5-Br-3-indolyl (with ferricyanide, phenazonium methosulfate or nitro BT and without any oxidation agent) were used an evaluated concerning the specificity, localization ability and inhibition of enzyme activity. Pretreatment of sections with distilled water or saline and inhibition by p-Cl-mercuribenzoate, glucono- and galactono-lactones were used for the characterization of the demonstrated enzyme activity.6-Br-2-naphthyl--D-glucoside is the most specific substrate for hetero--galactosidase. It is not split by lactase and acid -galactosidase. Only lysosomal -glucosidase can interfere. Because the latter enzyme is membrane-bound the difference in color intensity between untreated and prewashed sections are due to hetero--galactosidase. Only localization on the cellular (not intracellular) level can be achieved, however.The simultaneous azocoupling method with -naphthyl--D-glucoside and hexazonium-p-rosaniline enables a very good localization of hetero--galactosidase in the rabbit intestine. Due to a great inhibition exerted by hexazonium-p-rosaniline on the enzyme activity the method is unsuitable for the detection of hetero--galactosidase in zymograms and in the human intestine. Interference of lactase (or lactase-phlorizine hydrolase complex) is to be considered. The lysosomal -glucosidase does not seem to interfere.Indigogenic methods are not sensitive either. With ferricyanide as an oxidation agent it was not possible to detect the activity of hetero--galactosidase in zymograms and in sections. This is possibly due to overoxidation of indigo. The same holds true for phenazonium methosulfate used for the processing of zymograms. However, it was possible to reveal the activity of hetero--galactosidase in sections of the rabbit and monkey intestine with phenazonium methosulfate as oxidation agent. Nitro BT enhanced the coloration both in zymograms and in sections. In the latter case diffusion artifacts cannot be prevented, however. The interference of lactase, lysosomal -galactosidase and possibly of lysosomal -glucosidase (depending on the glycoside used) is always to be considered.Hetero--galactosidase was localized in the cytoplasm (particularly in the supranuclear region) of differentiated enterocytes covering the villi of the rabbit (the highest activity), monkey and human (the lowest activity) intestine. In crypt enterocytes and in cells of Brunner's glands the activity was lower. The occurrence of a low activity of hetero--galactosidase in the brush border of enterocytes of the rabbit intestine was also demonstrated.A proximodistal gradient was observed in the rabbit and monkey intestine, the upper jejunum displaying the highest activity.In jejunal biopsies of patients with celiac sprue (in the acute stage of the disease) the activity of hetero--galactosidase was lowered. No changes of activity were observed in jejunal biopsies of patients with isolated deficiencies of lactase or sucrase.In the rat kidney the enzyme was demonstrated particularly in the cytoplasm of cells of proximal convoluted tubules.     

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.     

Taxonomisch-chorologische Studie über die ArtLeucanthemum rotundifolium (W. K.) DC.

Die Arbeit befasst sich mit der Nomenklatur, Morphologie, Phytogeographie, Chorologie, Phytozönologie und Ökologie der ArtLeucanthemum rotundifolium (W. K.) DC. innerhalb ihres Gesamtareals. Auf Grund der Analyse aller untersuchten Faktoren werden die Endergebnisse vom taxonomischen Gesichtspunkte vorgelegt.L. rotundifolium wird mit der am nächsten verwandten ArtL. vulgare Lam. s. l. verglichen.     

Effect of root tip amputation on spiral oscillations of the growing hypocotyl with radicle of the pea (Pisum sativum L.)

In order to interpret the mechanism of elongation growth of the hypocotyl with radicle in the first stages of germination of the pea seed (Pisum sativum L.) a cybernetic model utilizing feed-back as a mechanism of correcting the wrong direction of growth was proposed (Spurný 1966, 1967). In the present study, the effect of amputation of the root tip as the control centre on the trajectory of the growing radicle was investigated. The results have shown that the hypocotyl grows, elongates—the rate of growth being slightly lower than that of the standard, but that no spiral oscillations at all are executed by the organ after amputation of the root tip. This finding appears to confirm the applicability of the proposed cybernetic model, for amputation of the root tip means that not only the control block is eliminated, but also that the channel of feed-back impulses to zone of elongation is interrupted.     

Ultrastructure of the surface of multiple scars inSaccharomycodes ludwigii

The use of induced primuline fluorescence led to the discovery of a new type of yeast scars (multiple scars) in the generaKloeckera, Saccharomycodes, Nadsonia andHanseniaspora. The structure and ultrastructure of their surface was studied by electron microscopy, using carbon replicas and isolated cell walls.     

Ribosomes in growing and non-growing bacterial cells

1. 1.
   It was found that growing cells always contained more different groups of ribosomes and their total amount was greater than in non-growing cells or spores, in which only two ribosome groups with different sedimentation constants, according to the species of microorganism, were consistently found.     

Contribution to the structural characterization of gamma globulin from pig colostrum

From the results obtained in the present work it is concluded that gamma globulin of the 7 S type (γG) which represents the main immunoglobulin component of pig colostrum, differs from serum γG globulin by the presence of another type of polypeptide chain, designed L₂; the latter was detected after S-sulfonation in starch gel electrophoresis as the fastest moving component and its immunoelectrophoretic pattern shows the presence of two precipitating components. Preparation of soluble heavy and light chains enabled us to study them imunochemically. The heavy chain is represented by two zones; the fainter one was not detected in comparative analysis of the heavy chain of serum γG globulin. The L₁ chain of colostral gamma globulin and the light chain of serum gamma globulin seem to contain three precipitating components, one of which appears due to aging of the chain solution in the presence of glycine. The material from colostrum seems to contain a larger amount of this component than serum. Further it was shown that the component arising in this way is antigenically closely related to the heavy chain.     

Study of the growth and development ofChlorella populations in the culture as a whole

Colour tests for study of the nuclear apparatus ofChlorella cells are described. It can be concluded from the results thatChlorella cell nucleus contains a relatively large nucleolus; the peripheral layers of the nucleus are rich in chromatin only in the nuclear division phase. The nucleus divides by a simplified form of mitosis, characterized morphologically by simple constriction.     

Die Anwendung einer ‘deduktiven Methode syntaxonomischer Klassifikation’ bei der Bearbeitung der strassenbegleitenden Pflanzengesellschaften Nordostböhmens

Zusammenfassung In der vorgelegten Arbeit wurde die sogenante deduktive Klassifikationsmethode der anthropogenen Pflanzengesellschaften behandelt. Diese Methode geht vom System der höheren Vegetationseinheiten der Braun-Blanquet Schule aus. Sie ermöglicht die syntaxonomische Bearbeitung zahlreicher anthropogener Gesellschaften unserer Kulturlandschaft, die nur aus Arten mit breiterer ökologischer und zönologischer Amplitude zusammengesetzt sind, und zwar ohne unerwünschte Verwechslung der Bestimmungsmerkmale einzelner Rangstufen des Systems (Klassen-, Ordnungs-, Verbands- und Assoziationskenn-und trennarten). Die Grundprinzipien der Methode wurden schon früher von Kopecký & Hejný (1971, 1973, 1974) und Kopecký (1976) erläutert. Die vorliegende Arbeit bringt eine präzisierte Auslegung der Methode auf Grund neuer Erfahrungen, die bei der Bearbeitung der spontan entstandenen strassenbegleitenden Gesellschaften derMolinio-Arrhenatheretea Tx. 1937 in Nordostböhmen gewonnen wurden. Aus dem reichen Aufnahmematerial werden nur einige Aufnahmen der Basalgesellschaften mitAlopecurus pratensis und der von ihnen abgeleiteten Derivatgesellschaften mitDactylis glomerata undAgropyron repens fur die anschauliche Demonstration der Methode beigegeben (Tab. 1 und 2). Die Aufnahmen der übrigen Gesellschafts-typen werden in einer anderen Arbeit (Kopecký 1978) veröffentlicht.

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Summary Use of a so-called deductive method in syntaxonomic classification of roadside plant communities in NE Bohemia.The present paper explains the so-called deductive method of syntaxonomic classification of plant communities, derived from the system of syntaxa of higher rank of the Braun-Blanquet approach. This method enables the synthesis of vegetations units of the varied anthropogenic plant communities in the man-made landscape, and their arrangement within the existing system, without changing the classification and determination criteria of both the higher and lower syntaxa in this system. Basic principles of this method have been outlined earlier by Kopecký & Hejný (1971, 1973, 1974) and Kopecký (1976). Following the newly obtained data and experience, this paper brings further details and more precise explanation of the method. Recently, the deductive method has been applied to the classification of turf communities which spontaneously develop along the roads in NE Bohemia. In order to illustrate the principles of the method, selected relevés of basal communities withAlopecurus pratensis (Table 1), and respective communities derived from these basal communities (Table 2) are presented. A complete set of vegetation relevés of all other roadside communities will be published elsewhere (Kopecký 1978).

Die wissenschaftlichen Pflanzennamen sind nach Rothmaler et al. (1966) angeführt.