Carbohydrate specificity of the receptor sites of mistletoe toxic lectin-I.

The carbohydrate specificity of mistletoe toxic lectin-I (ML-I) was studied by haemagglutination-inhibition assay. The results indicated that ML-I has a broad range of affinity for Gal alpha,beta linked sequences. The galabiose (E, Gal alpha 1----4Gal) sequence, a receptor of the uropathogenic E. coli ligand, was one of the best disaccharide inhibitors tested. The lectin also exhibits affinity for Lac(Gal beta 1----4Glc), T(Gal beta 1----3GalNAc), I/II(Gal beta 1----3/4GlcNAc) and B(Gal alpha 1----3Gal) sequences. Gal alpha 1----4Gal and Gal beta 1----4Glc are frequently occurring sequences of many glycosphingolipids located at the mammalian cell membranes, such as intestinal and red blood cell membranes, for ligand binding and toxin attachment. This finding provides important information concerning the possible mechanism of intoxication of cells by the mistletoe preparation.     

The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein.

The SEC17 gene of Saccharomyces cerevisiae is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus. Here we report that the product of the SEC17 gene has the exact biochemical properties expected for a yeast homologue of the mammalian transport factor, alpha-SNAP. The DNA sequence of SEC17 codes for a protein of predicted molecular mass of 33 kDa. Immunoblotting indicates that Sec17p fractionates as a peripheral membrane protein and is mostly soluble when overexpressed, suggesting the presence of a saturable membrane receptor for Sec17p. Sec17p was purified from yeast cytosol using a SNAP-dependent in vitro mammalian Golgi transport assay. Kinetic analysis using this assay shows Sec17p acts temporally close to the fusion of transport vesicles with the medial Golgi compartment. In yeast extracts, Sec17p binds to Sec18p with a 1:1 stoichiometry. The interaction between Sec17p and Sec18p requires an activity provided by yeast membranes, and this putative membrane receptor activity is not extracted by high salt treatment of membranes.     

Vertical profiles of CH4 concentrations,dissolved substrates and processes involved in CH4 production in a flooded Italian rice field

Vertical profiles were measured in soil cores taken from flooded rice fields in the Po valley during July and August 1990. Methane concentrations generally increased with depth and reached maximum values of 150–500 μM in 5–13 cm depth. However, the shape of the profiles was very different when studying different soil cores. The CH₄ content of gas bubbles showed a similar variability which apparently was due to spatial rather than temporal inhomogeneities. Similar inhomogeneities were observed in the vertical profiles of acetate, propionate, lactate, and formate which showed maximum values of 1500, 66, 135, and 153, μM, respectively. However, maxima and minima of the vertical profiles of the different substates usually coincided in one particular soil core. Large inhomogeneities in the vertical profiles were also observed for the rates of total CH₄ production, however, the percentage contribution of H₂/CO₂ to CH₄ production was relatively homogeneous at 24 ± 7% (SD). Similarly, the H₂ content of gas bubbles was relatively constant at 93.3 ± 9.6 ppmv when randomly sampled in the rice field at different times of the day. A small contribution (6%) of H₂/CO₂ to acetate production was also observed. Vertical profiles of the respiratory index (RI) for [2-¹⁴C] acetate showed that acetate was predominantly degraded by methanogenesis in 5–11 cm depth, but by respiration in the surface soil (3 cm depth) and in soil layers below 13–16 cm depth which coincided with a transition of the colour (grey to reddish) and the physical characteristics (porosity, density) of the soil. The observations indicate that the microbial community which degrades organic matter to CH₄ is in itself relatively homogenous, but operates at highly variable rates within the soil structure. Author for correspondence     

Chronopharmacokinetics and Cardiovascular Effects of Nifedipine

Orcadian phase dependency in pharmacokinetics and hemodynamic effects on blood pressure and heart rate of different galenic formulations of nifedipine (immediate-release, sustained-release, and i.v. solution) were studied in healthy subjects or in hypertensive patients. Pharmacokinetics of immediate-release but not sustained-release and i.v. nifedipine were dependent on time of day: immediate-release nifedipine had higher C_max (peak concentration) and shorter t_max (time-to-peak concentration) after morning than evening application, and bioavailibility in the evening was reduced by about 40%. Orcadian rhythm in estimated hepatic blood flow as determined by indocyanine green kinetics may contribute to these chronokinetics. A circadian time dependency was also found in nifedipine-induced effects on blood pressure and heart rate as monitored by 24-h ambulatory blood pressure measurements. In conclusion, the dose response relationship of oral nifedipine is influenced by the circadian organization of the cardiovascular system as well as by the galenic drug formulation.     

Neue Aspekte zur zugzeitlichen Frugivorie der Gartengrasmücke (Sylvia borin)

Zusammenfassung Freilandbeobachtungen und frühere Laboruntersuchungen (Berthold 1976) führten bisher zu einer widersprüchlichen Bewertung der zugzeitlichen Frugivorie bei der Gartengrasmücke. Deshalb wurden das Wahlverhalten gegenüber verschiedenen Beeren und Früchten in deren natürlichem Gehänge in einer Voliere geprüft sowie die Auswirkung unterschiedlich frugivorer Ernährung auf Körpermasse und tägliche Nahrungsaufnahme unter kontrollierten Laborbedingungen untersucht. Zwischen dem Präferenzverhalten gegenüber bestimmten Beeren und Früchten und deren Profitabilität bzw. Nährstoffgehalt bestand kein eindeutiger Zusammenhang. Demnach ist die Wahl zwischen verschiedenen Vegetabilien nicht einfach abhängig von ihrer Größe oder dem Nährstoffgehalt. Erhebliche Unterschiede zwischen den getesteten Beeren und Früchten zeigten sich bei der täglichen Aufnahmerate und den Auswirkungen auf die Körpermasse. Während bestimmte Beeren nur als Zusatzfutter zu einem immer, wenn auch geringfügig notwendigen, animalischen Standardfutter verabreicht werden konnten, waren die Gartengrasmücken mit Schwarzem Holunder und Feige auch bei ausschließlicher Gabe nicht nur in der Lage, ihre Körpermasse zu halten, sondern konnten dabei sogar normale Fettdeposition vollziehen. Offensichtlich sind bestimmte Vegetabilien für die Gartengrasmücke wesentlich bedeutsamer als bisher angenommen. Die Ergebnisse stimmen somit mit Beobachtungen aus dem Freiland überein. Die beobachteten qualitativen Unterschiede zwischen den Früchten und deren Wirkung auf die Körpermasse der Versuchsvögel ließen sich nicht allein mit dem Nährstoffgehalt der Beeren erklären. Vermutlich spielen die spezifische Qualität der Nährstoffe (z. B. Fettsäurezusammensetzung) und möglicherweise auch sekundäre Pflanzenstoffe eine wichtige Rolle.

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The significance of seasonal frugivory in migratory Garden WarblersSylvia borin

Summary In contrast to field observation, exhibiting an important role of seasonal frugivory,Berthold (1976) concluded that fruits are only a supplementary food, and he rejected the hypothesis that songbirds shift to a fruit diet during migration in order to render or to accelerate fat deposition.We investigated the preference of Garden Warblers upon various berries and pulpy fruits in an outdoor aviary. Furthermore, the effects of frugivory on body mass and food intake is measured in caged specimens under controlled laboratory conditions.In almost all feeding trials a significant selection between the two alternative fruit diets are obvious. However, no clearcut relationship between profitability or nutritive content of the various fruits and food choice is evident.In the experiments under controlled laboratory conditions the effects of varying extends of frugivory on body mass and food intake of captive Garden Warblers reveal a considerable difference between various berries and fruits. In some berries, the birds were not able to compensate for a reduction of the standard animal food by increasing frugivory. With Black Elder berries or figs, however, they did not only compensate for to maintain body mass rather they were able to gain mass during migratory fat deposition, even kept on an exclusive fruit diet.These results clearly exhibit a significant role of fruit diets in migrating Garden Warblers, with particular regards to Black Elder berries and figs, respectively.The observed differences between the various fruit diets do not correspond to the crude nutritive content of the pulp. Presumably, the specific quality of nutrients (for instance, fatty acid composition) or even secondary plant compounds have to be considered.The experimental results agree with field observations, indicating the importance of frugivory for the Garden Warbler, and they contradict Berthold's suggestion that fruits are only a least important supplementary food to an always necessary animal food.

     

Defects in fruiting body development caused by Tn5 lac insertions in Myxococcus xanthus.

Mutations caused by insertions of Tn5 lac that block development are rare. At least six of the eight mutations examined appeared to be regulatory. Three of these were found to disrupt social motility, suggesting a particular importance for this function. One other occurred in a known cell-cell interaction gene, bsgA, and the remaining two were located in genes operative early in the developmental program.     

Physical and chemical scavenging of singlet molecular oxygen by tocopherols

Singlet molecular oxygen (1O2) arising from the thermal decomposition of the endoperoxide of 3,3'-(1,4-naphthylidene) dipropionate was used to assess the effectiveness of alpha-, beta-, gamma-, and delta-tocopherol in the physical quenching as well as the chemical reaction of 1O2. The relative physical quenching efficiencies of the tocopherol homologs were found to decrease in the order of alpha greater than or equal to beta greater than gamma greater than delta-tocopherol. The ability of physical quenching depends on a free hydroxyl group in position 6 of the chromane ring. Chemical reactivity of the tocopherol homologs with 1O2 was low, accounting for 0.1-1.5% of physical quenching with beta-tocopherol showing particularly low reactivity, resulting in the sequence alpha greater than gamma greater than delta greater than beta-tocopherol. Tocopheryl quinones were products of all tocopherol homologs, and in addition a quinone epoxide was a major product from gamma-tocopherol. This quinone epoxide was not cleaved by rat liver microsomal epoxide hydrolase; however, it reacted further with 1O2. It is concluded that methylation in position 5 of the chromane ring enhances physical quenching of 1O2, whereas chemical reactivity is favored by a methylated position 7. In view of the fact that beta-tocopherol is as effective as alpha-tocopherol in physical quenching of 1O2 but shows very low chemical reactivity, this tocopherol homolog might be particularly suitable for biological conditions in which an accumulation of oxidation products might weaken the antioxidant defense.     

Avian alcohol dehydrogenase. Characterization of the quail enzyme, functional interpretations, and relationships to the different classes of mammalian alcohol dehydrogenase

The primary structure of the major quail liver alcohol dehydrogenase was determined. It is a long-chain, zinc-containing alcohol dehydrogenase of the type occurring also in mammals and hence allows judgement of the gene duplications giving rise to the classes of the human alcohol dehydrogenase system. The avian form is most closely related to the class I mammalian enzyme (72-75% residue identity), least related to class II (60% identity), and intermediately related to class III (64-65% identity). This pattern distinguishes the mammalian enzyme classes and separates classes I and II in particular. In addition to the generally larger similarities with class I, the avian enzyme exhibits certain residue patterns otherwise typical of the other classes, including an extra Trp residue, present in both class II and III but not in class I, with a corresponding increase in the UV absorbance. The avian enzyme further shows that a Gly residue at position 260 previously considered strictly conserved in alcohol dehydrogenases can be exchanged with Lys. However, zinc-binding residues, coenzyme-binding residues, and to a large extent substrate-binding residues are unchanged in the avian enzyme, suggesting its functional properties to be related to those of the class I mammalian alcohol dehydrogenases. In contrast, the areas of subunit interactions in the dimers differ substantially. These results show that (a) the vertebrate enzyme classes are of distant origin, (b) the submammalian enzyme exhibits partly mixed properties in relation to the classes, and (c) the three mammalian enzyme classes are not as equidistantly related as initially apparent but suggest origins from two sublevels.     

Synthesis and structural stability of helichrome as an artificial hemeproteins

A detailed procedure is described for the synthesis of helichrome, which is the first successful example of polypeptide-based artificial hemeprotein. The segment synthesis-condensation approach used for the assembly of small proteins has proven to be extremely useful for protein mimetics as well. The final deprotection was performed using the TMSOTf-thioanisole method instead of the less-convenient hydrogen fluoride method. The unfolding transition of the alpha-helical conformation of helichrome induced by guanidine hydrochloride was studied to understand the stability and dynamics of the folded structure. The resulting parameters (C0.5 = 5.2 M and delta GH2O = -4.4 kcal mol-1) characterizing helichrome denaturation were comparable to that of native globular proteins.