Tetrazolium [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction by mycoplasmas.

We investigated 22 mycoplasma and acholeplasma species for their ability to reduce tetrazolium salts by using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The test results were evaluated visually, as well as spectrophotometrically, by using an enzyme-linked immunosorbent assay reader. Our results were very similar to the results obtained when the tetrazolium salt reduction assay described by Aluotto et al. was used. However, the MTT reduction assay appeared to be better because it is faster, more objective and sensitive, easier to evaluate, and less expensive; in addition, it allows quantitative determinations. By using regression analysis a linear correlation between formazan production and the number of colony-forming units was demonstrated for all of the species investigated, indicating that the MTT assay can also be used for growth, toxicity, or chemosensitivity tests for the mycoplasma species that are capable of reducing tetrazolium salts.     

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 human MUC2 intestinal mucin has cysteine-rich subdomains located both upstream and downstream of its central repetitive region.

The human MUC2 mucin is a large secretory glycoconjugate that coats the epithelia of the intestines, airways, and other mucus membrane-containing organs. Previous work has shown that this mucin contains an extended tandem repeat-containing domain rich in Thr and Pro. In the present work we describe two additional regions of this mucin located both upstream and downstream of the tandem repeat array. The carboxyl-terminal domain contains 984 residues and can be divided into mucin-like (139 residues) and cysteine-rich (845 residues) subdomains. This latter subdomain exhibits varying degrees of sequence similarity to a wide range of mucins and mucin-like proteins including those isolated from rats, pigs, cows, and frogs. We also report here the sequence of 1270 residues lying immediately upstream of the tandem repeats. This region contains a repetitive, mucin-like subdomain and a second cysteine-rich stretch of more than 700 residues. Both cysteine-rich subdomains of this mucin have sequence similarity with von Willebrand factor, a serum protein that exists as a disulfide-linked polymer. This suggests that these cysteine-rich subdomains are important in the catenation of mucin monomers into oligomers, the structures that confer viscoelasticity upon mucus.     

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     

Conformation and formation tendency of the cyclotetrapeptide cyclo (D-Pro-D-Pro-L-Pro-L-Pro): experimental results and molecular modeling studies

The title compound represents the smallest member of cyclic proline peptides corresponding to the general formula c(DDLL-Pro4)n with a strictly D,D,L,L double-alternating sequence of the chiral amino acid residues. The cyclopeptides with n greater than or equal to 2 could be synthesized from both DDLL-Pro4 (1) and DLLD-Pro4 (2). The cyclic monomer (n = 1) resulted only from 2, whereas not even a trace could be found by cyclization of 1. The peptide exists in a strongly strained Ci symmetrical conformation (x-ray analysis) with alternating cis and trans peptide bonds (ctct form I). The cis peptide bonds deviate from planarity (omega = 22 degrees); two of the pyrrolidine rings show a "South" conformation (phi = -94 degrees), whereas the other residues exhibit C alpha-endo puckering (phi = -124 degrees). Two of the psi angles surprisingly occur at +41 degrees (anti-cis'), the others are located in the trans' region. A quantitative ring opening occurs with trifluoroacetic acid at room temperature. In solution the existence of an isomeric ctcc sequence (form Ia) is indicated. Dreiding model studies also suggested a favorable conformation with a tctc sequence (form II). Consequently, we performed molecular mechanics calculations, based on the CHARMM force field and semiempirical quantum mechanical AM1 calculations (MOPAC program). Pronounced differences in the backbone parameters were found using these two methods. However, the theoretical studies evidenced the experimentally obtained differences in the cyclization tendencies of the linear precursors.     

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.

     

Reflex control of vascular capacitance during hypoxia, hypercapnia, or hypoxic hypercapnia

We tested the hypothesis that the changes in venous tone induced by changes in arterial blood oxygen or carbon dioxide require intact cardiovascular reflexes. Mongrel dogs were anesthetized with sodium pentobarbital and paralyzed with veruronium bromide. Cardiac output and central blood volume were measured by indocyanine green dilution. Mean circulatory filling pressure, an index of venous tone at constant blood volume, was estimated from the central venous pressure during transient electrical fibrillation of the heart. With intact reflexes, hypoxia (arterial PaO2 = 38 mmHg), hypercapnia (PaCO2 = 72 mmHg), or hypoxic hypercapnia (PaO2 = 41; PaCO2 = 69 mmHg) (1 mmHg = 133.32 Pa) significantly increased the mean circulatory filling pressure and cardiac output. Hypoxia, but not normoxic hypercapnia, increased the mean systemic arterial pressure and maintained the control level of total peripheral resistance. With reflexes blocked with hexamethonium and atropine, systemic arterial pressure supported with a constant infusion of norepinephrine, and the mean circulatory filling pressure restored toward control with 5 mL/kg blood, each experimental gas mixture caused a decrease in total peripheral resistance and arterial pressure, while the mean circulatory filling pressure and cardiac output were unchanged or increased slightly. We conclude that hypoxia, hypercapnia, and hypoxic hypercapnia have little direct influence on vascular capacitance, but with reflexes intact, there is a significant reflex increase in mean circulatory filling pressure.     

A sequence-specific protease cleaves a maternal, cortical protein during early embryogenesis in Sciara coprophila (Diptera)

One of the first events of egg activation in Sciara coprophila (Diptera) is the disappearance of an abundant maternal 38-kDa protein (p38) and the simultaneous emergence of an abundant 35-kDa protein (p35). Western blotting experiments using monoclonal antibodies directed against p38 reveal that p38 and p35 are serologically related and indicate that maternal p38 is transformed into p35 during early development. This transition is possibly accompanied by a conformational change in the part of the protein that is common to both protein species. The processing of p38 to p35 can be mimicked by trypsin treatment in vitro, suggesting that a trypsin-like protease is responsible for this conversion in vivo. Immunostaining indicates that the p38 class of antigens is evently distributed in the periplasm of early cleavage embryos. After the arrival of nuclei in the periplasm, the antigens become associated with the infolding cellular membranes. A similar membrane association of actin can be observed with anti-actin antibodies. Nevertheless, p38 and actin are clearly distinct from each other. We presume that p38 is a product of a maternal effect gene necessary for early dipteran development.     

Adaptation of microbial activities to the environmental conditions in alpine soils

Summary The adaptation of soil microorganisms to different environmental conditions was investigated in the Austrian Central Alps (Hohe. Tauern). The floristic composition of the soil fungi at different sites was determined and the CO₂-release from soils taken from different altitudes was measured at different temperatures. The results showed a decreasing diversity of soil fungi with increasing altitude and a change in the dominating species at different altitudes and/or with vegetation patterns. The relative rates of CO₂-release from soils from different altitudes did not differ at different incubation temperatures. It was concluded that, among soil fungi the selection of species is a more effective mechanism for the adaption to changed environmental conditions than metabolic adaptations.This study was supported in part by the Österreichische MaB-Hochgebirgsprogramm Hohe TauernDedicated to Dr. K.F. Springer