Evidence for a fucose-binding protein in boar spermatozoa

A fucose binding protein was detected in boar spermatozoa by means of a specifically developed modified enzyme-linked-lectin-assay using glycosylated peroxidase derivatives. The distribution of the fucose binding protein was assessed by means of fluorescence microscopy with fluoresceinyl-glycosylated peroxidase. Fucose binding was particularly prominent at the apical region of the sperm head. In order to gain more insight into the precise localization of the carbohydrate binding protein electron microscopical studies were performed using fucosyl peroxidase coupled to colloidal gold. In ultrathin sections as well as in specimens prepared in toto for TEM an intensive binding of fucosylperoxidase-colloidal gold was predominantly found at the apical part of the acrosome appearing as a crescent-like area. In some cases this binding pattern was replaced by a triangle-like intensive labelling at the equatorial segment as revealed clearly by specimens prepared in toto. By SDS-PAGE of the SDS-extractable sperm-proteins, followed by transblotting to nitrocellulose and visualization with the fucosylperoxidase by enzymatic amplification with 4-chloro-1-naphthol mainly one protein with the reduced molecular weight of approximately 53 kdal and some small proteins with apparent molecular weights less than 20 kdal was found to be responsible for the fucose-binding ability of porcine spermatozoa.     

Evidence for a fucose-binding protein in boar spermatozoa

Summary A fucose binding, protein was detected in boar spermatozoa by means of a specifically developed modified enzyme-linked-lectin-assay using glycosylated peroxidase derivatives. The distribution of the fucose binding protein was assessed by means of fluorescence microscopy with fluoresceinyl-glycosylated peroxidase. Fucose binding was particularly prominent at the apical region of the sperm head. In order to gain more insight into the precise localization of the carbohydrate binding, protein electron microscopical studies were performed using fucosyl peroxidase coupled to colloidal gold. In ultrathin sections as well as in specimens, prepared in toto for TEM an intensive binding of fucosylperoxidase-colloidal gold was predominantly found at the apical part of the acrosome appearing as a crescent-like area. In some cases this binding pattern was replaced by a triangle-like intensive labelling at the equatorial segment as revealed clearly by specimens prepared in toto. By SDS-PAGE of the SDS-extractable sperm-proteins, followed by transblotting to nitrocellulose and visualization with the fucosylperoxidase by enzymatic amplification with 4-chloro-1-naphthol mainly one protein with the reduced molecular weight of approximately 53 kdal and some small proteins with apparent molecular weights less than 20 kdal was found to be responsible for the fucose-binding ability of porcine spermatozoa.     

Fluoreszenzmarkierung von Tumor- und Leber-DNS der Ratte

Zusammenfassung Es wird eine Methode zur Fluoreszenzmarkierung von DNS, die aus Guérin-Rattentumoren und Rattenlebern isoliert wurde, mitgeteilt. Zur Kopplung wurde 1-Dimethylamino-naphthalin-sulfochlorid-5 (DIS) verwendet. Die Ratten intravenös applizierte markierte DNS stellt sich im Gewebsschnitt durch ihre gelbe bzw. intensiv gelbgrüne Fluoreszenz dar.

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Summary This paper informs of a method of fluorescence marking of DNA isolated from Guérin rat tumors and rat livers. For coupling we used 1-dimethylaminonaphthalene-sulphochloride-5 (DIS). The marked DNA injected intravenously into rats presented itself by yellow or intense yellowish green fluorescence.

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Zum hundertjährigen Bestehen des Lehrstuhls für Pathologie der Universität Rostock.     

The xylem sap of maple (Acer platanoides) trees--sap obtained by a novel method shows changes with season and height

Xylem sap of log pieces of maple trees was collected by a novelpressure/decompression method developed recently for the mechanicaldrying of timber. Seasonal changes and spatial distributionsof the osmotic potential, the pH and the concentrations of majorsolutes and of the plant stress-hormone abscisic acid (ABA)were investigated. Sucrose and quebrachitol were the main components of the xylemsap. The sucrose concentration varied between 10 mM and 25 mMduring the winter months and declined to a minimum during theperiod of bud burst. Quebrachitol was found in concentrationsof up to 7 mM with a high variability throughout the year. Highconcentrations of ABA were measured during the summer seasonand in mid-winter. Rainfall caused an increase of ABA in somesamples. The osmotic potential of the xylem sap increased with the heightof the collection site. The pH of the sap decreased by approximatelyone unit between the base of the trunk and the crown. The increaseof the osmotic potential was mainly due to sucrose, quebrachitoland potassium. Malate contributed to the decrease of the pH.ABA of the xylem sap increased with decreasing moisture contentof the wood. Key words: Pressure/decompression method, xylem sap, abscisic acid, sucrose, quebrachitol, Acer platanoides     

Composition of Orchid Scents Attracting Euglossine Bees

Fragrance composition of flowers from 101 plant species, especially orchids, were analyzed. Several compounds, including allo-aromadendrene, β-bourbonene, α-copaene, α-cubebene, 1,2-dimethoxybenzene, 1,3,5-trimethoxybenzene, epoxygeranyl acetate, 7,11-epoxymegastigma-5(6)-en-9-one, two γ-lactones, germacradienol, germacrene D, humulene, methoxy-phenyl-ethyl acetate, myrcene epoxide, sabinene, styrene, and undecatriene were detected for the first time in orchids. Fragrance composition of flowers pollinated by male euglossine bee species (perfume flowers) of four plant families are compared and contrasted with those of orchid species with other pollination systems. Melittophilous, but highly specialized orchids, produce fragrances rich in different sesquiterpenes and other rare compounds. In the species that are exclusively pollinated by fragrance-collecting male euglossine bees, the fragrances are primary attractants that serve both as an attractant and as a reward. The unusually intensely smelling flowers mostly produce esters and monoterpenes. The fragrances of euglossophilous flowers of the three plant families investigated are composed of nearly the same sets of chemical compounds, suggesting convergent evolution. Typically, euglossophilous plant species produce large amounts of few fragrant substances while melittophilous species often produce rare compounds in small amounts.     

Electron microscopic localization of a fucose-binding protein in acrosome reacted boar spermatozoa by the fucosyl-peroxidase-gold method

In this study we have examined the behaviour and the localization of the fucose-binding protein (FBP) in boar spermatozoa during ionophore induced acrosome reaction (AR) by means of normal TEM and specimen preparation in toto. During early stages of AR the FBP is first localized at the border between equatorial segment and anterior acrosome. With the propagation of the AR the FBP is dramatically expressed and visible over the entire surface of the acrosome and equatorial segment. TEM pictures of this stages show that the FBP is associated with the OAM. At later stages of AR, when acrosomal ghost formation occur, the FBP is associated with the acrosomal ghost, and equatorial segment and to a very low degree also with the IAM. It is concluded from this data that the FBP is responsible for the specific binding of the ghost-sperm unit to the zona pellucida.     

Effect of autovaccination on the course of urinary tract infection in animal experiment

Autovaccination of rats with chronic pyelonephritis carried out approximately two months after the onset of infection does not result in an improved histological picture in the infected but can prevent destructive processes in the controlateral kidney. Cyclophosphamide administered in three doses of 30 mg/kg simultaneously with autovaccination slightly modulates the immune response to the infectious strain. The temporal relationship between immunization and cyclophosphamide administration determines the mode of action of cyclophosphamide. In the present experiment, the concept of Miller has not proved to be applicable. Cyclophosphamide administration causes a distinct increase in inflammatory processes in the kidney. Should an enhancement phenomenon be involved in the bacterial infection of the kidney, of which we have found no proof, cyclophosphamide therapy as it was used in the present study would not result in its removal and thus in improved elimination of the infectious organism. Additional experiments are required to determine whether animals subjected to autovaccination are protected against a new episode of urinary tract infection.     

Reduction of molecular gas diffusion through gaskets in leaf gas exchange cuvettes by leaf‐mediated pores

There is an ongoing debate on how to correct leaf gas exchange measurements for the unavoidable diffusion leakage that occurs when measurements are done in non‐ambient CO₂ concentrations. In this study, we present a theory on how the CO₂ diffusion gradient over the gasket is affected by leaf‐mediated pores (LMP) and how LMP reduce diffusive exchange across the gaskets. Recent discussions have so far neglected the processes in the quasi‐laminar boundary layer around the gasket. Counter intuitively, LMP reduce the leakage through gaskets, which can be explained by assuming that the boundary layer at the exterior of the cuvette is enriched with air from the inside of the cuvette. The effect can thus be reduced by reducing the boundary layer thickness. The theory clarifies conflicting results from earlier studies. We developed leaf adaptor frames that eliminate LMP during measurements on delicate plant material such as grass leaves with circular cross section, and the effectiveness is shown with respiration measurements on a harp of Deschampsia flexuosa leaves. We conclude that the best solution for measurements with portable photosynthesis systems is to avoid LMP rather than trying to correct for the effects.