A rapid colorimetric assay for heparinase activity.

A rapid, sensitive, assay for enzymes that degrade heparin is described. The procedure is based on the interference of heparin with color development during the interaction of protein with the dye Coomassie brilliant blue. The loss of this property when the glycosaminoglycan is degraded by heparinase can be used to quantify activity of the enzyme in pure form, or in complex biological samples such as tissue homogenates or serum. The assay is also suitable for studying dependence of heparinase activity under conditions such as varying pH and temperature.     

Flora von Oesterreich-Ungarn

Ohne Zusammenfassung     

Sulphamerazine toxicity in cut-throat trout broodfish Salmo clarki (Richardson)

Treatment of cut-throat trout broodfish Salmo clarki (Richardson) with Sulphamerazine at 220 mg/kg (10 g/100 1b) of fish/day for 14 days resulted in severe kidney histopathology and increased mortality among males. Experimental data presented showed that cut-throat trout broodfish were extremely sensitive to Sulphamerazine toxicity. Hydropic degeneration of renal tubule epithelium and haemorrhage into tubule lumens were observed in kidneys of both male and female trout, but was more severe in the former. Death, which occurred only in males, was correlated with spawning stress and impaired renal function.     

Feasible mechanisms for algal digestion in the king angelfish

To determine the ability of the king angelfish Holacanthus passer to digest algae, three algal species were immersed in acidic conditions similar to that found in the stomach of fish. Only one of them was not susceptible to acidic lysis; two were affected after 40 and 60 min at pH 2·0. King angelfish have an expanded region of the intestine called here the hindgut chamber (HC) containing populations of micro-organisms. Some of these micro-organisms have the capacity to grow in cellulose, agar, and alginic acid; the main components of algal cell walls. Micro-organisms grew in carboxymethylcellulose cultures under aerobic and micro-aerobic conditions. The HC is highly vascularized, which could increase absorptive efficiency of material digested in it.     

Heat treatment results in a loss of transgene-encoded activities in several tobacco lines.

Heat treatment (37 degrees C) of transgenic tobacco (Nicotiana tabacum) plants led to a reversible reduction or complete loss of transgene-encoded activities in about 40% of 10 independent transformants carrying the luciferase-coding region fused to the 355 cauliflower mosaic virus or the soybean small subunit promoter and the nopaline synthase promoter driving the neomycin phosphotransferase gene, whereas the other lines had temperature-tolerant activities. Temperature sensitivity or tolerance of transgene-encoded activities was heritable. In some of the lines, temperature sensitivity of the transgene-encoded activities depended on the stage of development, occurring in either seedlings (40% luciferase and 50% neomycin phosphotransferase) or adult plants (both 40%). The phenomenon did not correlate with copy numbers or the homo- or hemizygous state of the transgenes. In lines harboring a temperature-sensitive luciferase activity, reduction of bioluminescence was observed after 2 to 3 h at 37 degrees C. Activity was regained after 2 h of subsequent cultivation at 25 degrees C. Irrespective of the reaction to the heat treatment, the level of luciferase RNA was slightly increased at 37 degrees C. Only in lines showing temperature sensitivity of transgene-encoded activities was the amount of luciferase and neomycin phosphotransferase strongly reduced. In sterile culture, heat treatment for 15 d did not cause visible damage or changes in plant morphology. In all plants tested a slight induction of the heat-shock response was observed at 37 degrees C.     

Externally attached radio transmitters do not affect the parental care behaviour of rock bass

Turning, pectoral fin and caudal fin rates and time spent on the nest of male rock bass Ambloplites rupestris , engaged in parental care, were not affected after the attachment of external radio transmitters. Reproductive success was similar between treatment and control fish. Micro external radio transmitters can be used on small fishes for studying parental care duration and post-care movement without altering their behaviour.     

Interactions between two catalytically distinct MCM subgroups are essential for coordinated ATP hydrolysis and DNA replication.

The six MCM (minichromosome maintenance) proteins are essential DNA replication factors that each contain a putative ATP binding motif and together form a heterohexameric complex. We show that these motifs are required for viability in vivo and coordinated ATP hydrolysis in vitro. Mutational analysis discriminates between two functionally distinct MCM protein subgroups: Mcm4p, 6p, and 7p contribute canonical ATP binding motifs essential for catalysis, whereas the related motifs in Mcm2p, 3p, and 5p serve a regulatory function. Reconstitution experiments indicate that specific functional interactions between these two subgroups are required for robust ATP hydrolysis. Our observations show parallels between the MCM complex and the F1-ATPase, and we discuss how ATP hydrolysis by the MCM complex might be coupled to DNA strand separation.