Enzymes of the jejunal enterochromaffine cells in man

Summary A study of enzymatic equipment of enterochromaffine cells (e.c.) in jenual biopsies obtained with a Crosby capsule in normal humans and patients with nontropical sprue was undertaken. The following enzymes were demonstrated: alkaline phosphatase and adenosine triphosphatase (cell membrane), acid phosphatase (corpuscular), non-specific esterase (diffuse and corpuscular, predominantly eserine resistant, in corpuscular localization E 600 resistant), DPN- and TPN-diaphorases and dehydrogenases of lactic acid, malic acid, isocitric acid, glucoso-6-phosphoric acid, succinic acid, -hydroxybutyric acid and -glycerophosphoric acid. Enzyme activities were not equal in all cells suggesting some type of secretory cycle. In most patients with untreated nontropical sprue or with the disease in relapse e.c. were more numerous and hypertrophic with elevated activities of non-specific esterase and acid phosphatase. Implications of these results are briefly discussed.With 8 Figures in the Text, of which 2 in Colour     

Factors affecting the accuracy of chlorophylla andb determination by means of their paper chromatographic separation and colorimetric measurement in eluates

Quantitative determination of chlorophyll a and β can be made by paper chromatography of acetone extracts of plant material with colorimetric measurement of the eluates from the separated zones. From the suitable solvent systems which give adequate separation of the pigments at a distance of 20 cm. from the start,Hager's mixture (1955) separates the chlorophylls better than the toluene-isopropanol (400: 1 v/v.) mixture, which, however, is better for the separation of carotenoids. Twice the amount of chlorophyll is separated on Whatman 31 ET paper, equally well and with the same time of development, as on Whatman No. 3 paper, on which it is possible to separate a maximum of about 15 μg of chlorophyll pigments per 1 em. start length. Losses on elution are, however, higher on using Whatman 31 ET paper. In plants with a high chlorophyllase activity, the error of determining chlorophyll a andb is greatly reduced if the leaves are placed for 1 min. in boiling water before extraction. For elution of chlorophylla andb from paper it is better to use anhydrous acetone, for chlorophyllides 80% acetone. A comparison of the procedure investigated with the method of two-wave length spectrophotometric measurement of crude acetone extracts showed that in view of the average 10% loss, the chromatographic method is hardly suitable for determining the absolute amounts of chlorophylla andb, although the relation (a/b) can be determined with similar precision by both methods. Moreover, in view of the greater amount of work involved the chromatographic method can only be recommended for confirming the results of spectrophotometrie determination. Quantitative determination of chlorophylls from the area of the spot or from the "R_F" value can only be of an informative character.     

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.     

Corticotropin-Releasing Hormone Affects Short Immobilization Stress-Induced Changes in Lung Cytosolic and Membrane Glucocorticoid Binding Sites

Glucocorticoids act via glucocorticoid receptors (GR), typically localized in the cytosol (cGR). Rapid action is probably mediated via membrane receptors (mGR). In corticotropin-releasing hormone knockouts (CRH-KO), basal plasma glucocorticoid levels do differ from wild type levels (WT), but are approximately ten times lower during exposure to immobilization stress (IMMO) in comparison to WT. We tested the following hypotheses: (1) the mice lung tissue GR basal numbers would not be changed in CRH-KO (because of similar glucocorticoid levels), (2) the number of GR would be changed in WT but not in KO during short (30, 90, and 120 min) IMMO (because of higher increase of glucocorticoid levels in WT). The basal levels of cGR were not changed in CRH-KO (compared to WT), while mGR were significantly lower (62 %) in CRH-KO. In WT, there was the only decrease (to 32 %) in cGR after 120 min when we also found an increase in mGR in WT (to 201 %). In CRH-KO, IMMO caused gradual decrease in cGR (to 52 % after 30 min, to 46 % after 90 min, and to 32 % after 120 min). In CRH-KO, the only increase in mGR appeared already at 30 min of IMMO. These data suggest, on the contrary to our hypotheses, that CRH-KO are more susceptible to GR changes in early phases of stress.     

Improved localization of thiamine pyrophosphatase activity in mounted cryostat sections using gel fixation and gel incubation media

Summary The presence of 1% agar in the fixation and substrate solutions for the histochemical demonstration of thiamine pyrophosphatase (4.4 mM TPP; 3.6 mM Pb²⁺; 0.025 Tris-maleate buffer, pH 7.2) clearly facilitates the localization of the enzyme in Golgi apparatus in cold microtome sections prepared from unfixed specimens.     

Isolation and characterization of wheat peroxidase isoenzyme B1

Two pure peroxidase isoenzymes B1 and D4 were isolated from the upper parts of 10-day-old wheat seedlings by means of gel and ion-exchange chromatography. Their MWs were 85000 and 24000 respectively. B1 was unstable and under various conditions it was converted to another isoenzyme, electrophoretically identical with D4. B1 contains about 40% of neutral sugars: 17.2% arabinose, 15.3% galactose, 5% glucose and traces of mannose. D4 is free of neutral sugars. None of the isoenzymes contained amino sugars. B1 oxidizes ferulic and p-coumaric acids. This oxidation has two pH optima of 4.4 and 5.4–5.6 and is inhibited by high concentrations of substrates, cyanide and azide. B1 oxidizes IAA in the presence of phenolic cofactor and Mn²⁺ ions. IAA oxidation has two pH optima of 4.5 and 5.6 and is inhibited by high substrate concentration, cyanide and azide, and by a number of indole derivatives. The main products of IAA oxidation are 3-methyleneoxindole and indole-3-methanol. o- and p- diphenols induce a lag period prior to IAA oxidation. Ferulic acid is oxidized during this lag period, probably to a dimer. B1 is able to produce H₂O₂ from oxygen. Mn²⁺ ions, a phenolic cofactor and an electron donor (IAA or NADH) are needed. B1 oxidizes α-keto-γ- methylmercaptobutyric acid to ethylene. D4 has a low peroxidatic activity and is inactive as an IAA oxidase. Thus B1 is probably an active cell wall-bound peroxidase isoenzyme, whereas D4 is its decomposition product.     

Habitat requirements, short-term population dynamics and coexistence of native and invasive Impatiens species: a field study

The genus Impatiens (Balsaminaceae) includes three widespread species in the Czech Republic, central Europe: the native I. noli-tangere, and two invasive species, I. parviflora and I. glandulifera, differing in the dynamics of invasion. They all occur in similar habitats and share basic life-history characteristics, which make them a suitable model for studying species traits associated with invasiveness. In this study we investigated differences in habitat requirements of these Impatiens species, their coexistence and short-term population dynamics in the field. We established 84 1 × 1 m permanent plots in five localities where all three species co-occurred. In each plot vascular plant species were determined, their cover estimated and all individuals of Impatiens species counted. Site characteristics including tree canopy cover, soil moisture, nitrogen and carbon content, and slope were measured directly. Nutrients, light, humidity and soil reaction were estimated using Ellenberg indicator values. The presence of I. noli-tangere was strongly correlated with high soil moisture, that of I. parviflora with high tree canopy cover and low soil moisture. Impatiens glandulifera exhibited a unimodal response to tree canopy cover, avoiding both very shaded and fully open sites. The current-year abundances of all species were negatively related to those of congeneric species. These results suggest that the coexistence of Impatiens species in the same habitat is due to microsite differentiation. Further spread of I. glandulifera to new habitats, and reduction of the native I. noli-tangere niche, can be expected in areas where the latter species co-occurs with competitively strong invasive congeners.     

Production of microsporidia pathogenic to the Colorado potato beetle (Leptinotarsa decemlineata) in alternate hosts

The microsporida Nosema gastroideae and N. equestris, which are highly pathogenic for Leptinotarsa, have been successfully produced in some other chrysomelid species, Gastrophysa polygoni and G. viridula. As the principal target host, Leptinotarsa is very susceptible to these pathogens, and death occurs before massive sporulation by the microsporidia. By contrast, the infected larvae of G. polygoni or G. viridula are able to develop until the adult stage when most of the tissues become filled with spores. In addition, the larvae and adults of these species can be reared in the laboratory on Polygonum aviculare and Rumex obtusifolius. These plants have longer vegetative periods and are better sources of food than potato leaves. In both species of Gastrophysa the yields of spores related to unit weight were about five times higher than in Leptinotarsa. In the adults of G. viridula there was up to 4.8 × 10⁶ spores mg^?1 body weight of N. gastroideae, or 9.1 × 10⁶ spores mg^?1 of N. equestris. The higher content of microsporidian spores facilitates their purification and isolation.     

Using dot blot with immunochemical detection to evaluate global changes in SUMO-2/3 conjugation

Here we describe a non-invasive method for rapid and highly reproducible genotyping of transgenic mammals with ubiquitous expression of fluorophore reporters. Hair samples from transgenic mice and pigs with systemic expression of the fluorophore reporter Venus were analyzed with a fluorescence microscope in few minutes. The hair samples can be preserved for long-term storage at ambient temperature conditions. This non-invasive method is useful for genotyping of transgenic large animals and contributes to animal welfare by reducing stress and discomfort of the animals during sample collection.