Escherichia coli cells lacking methylation-blocking factor (leucine-responsive regulatory protein) have precise timing of initiation of DNA replication in the cell cycle.

A protein that is required for specific methylation inhibition of two GATC sites in the papBA pilin promoter region, known as methylation-blocking factor (Mbf) and recently shown to be identical to the leucine-responsive regulatory protein (Lrp), is not responsible for the delayed methylation at oriC implicated in an eclipse period following initiation of DNA replication. Cells containing a transposon mutation within the mbf (lrp) gene initiate DNA replication at the correct time during the cell cycle, whereas cells with increased amounts of the Dam methyltransferase initiate DNA replication randomly throughout the cell cycle.     

Cytotoxic activity of tumor necrosis factor is mediated by early damage of mitochondrial functions. Evidence for the involvement of mitochondrial radical generation.

Structural mitochondrial damage accompanies the cytotoxic effects of several drugs including tumor necrosis factor (TNF). Using various inhibitors of mitochondrial electron transport we have investigated the mechanism of TNF-mediated cytotoxicity in L929 and WEHI 164 clone 13 mouse fibrosarcoma cells. Inhibitors with different sites of action modulated TNF cytotoxicity, however, with contrasting effects on final cell viability. Inhibition of mitochondrial electron transport at complex III (cytochrome c reductase) by antimycin A resulted in a marked potentiation of TNF-mediated injury. In contrast, when the electron flow to ubiquinone was blocked, either at complex I (NADH-ubiquinone oxidoreductase) with amytal or at complex II (succinate-ubiquinone reductase) with thenoyltrifluoroacetone, cells were markedly protected against TNF cytotoxicity. Neither uncouplers nor inhibitors of oxidative phosphorylation nor complex IV (cytochrome c oxidase) inhibitors significantly interfered with TNF-mediated effects, ruling out the involvement of energy-coupled phenomena. In addition, the toxic effects of TNF were counteracted by the addition of antioxidants and iron chelators. Furthermore, we analyzed the direct effect of TNF on mitochondrial morphology and functions. Treatment of L929 cells with TNF led to an early degeneration of the mitochondrial ultrastructure without any pronounced damage of other cellular organelles. Analysis of the mitochondrial electron flow revealed that TNF treatment led to a rapid inhibition of the mitochondria to oxidize succinate and NADH-linked substrates. The inhibition of electron transport was dose-dependent and became readily detectable 60 min after the start of TNF treatment, thus preceding the onset of cell death by at least 3-6 h. In contrast, only minor effects were observed on complex IV activity. The different effects observed with the mitochondrial respiratory chain inhibitors provide suggestive evidence that mitochondrial production of oxygen radicals mainly generated at the ubisemiquinone site is a causal mechanism of TNF cytotoxicity. This conclusion is further supported by the protective effect of antioxidants as well as the selective pattern of damage of mitochondrial chain components and characteristic alterations of the mitochondrial ultrastructure.     

How predatory mites cope with the web of their tetranychid prey: a functional view on dorsal chaetotaxy in the Phytoseiidae

It has been proposed from behavioural observations that the ability of Phytoseiidae to penetrate the web is related to chaetotaxy. As phytoseiid species differ widely in chaetotaxy, as well as in their ability to cope with complex webs, this hypothesis was tested by studying relations between these two variables. In this paper we develop a simple method to quantify the degree to which the soma is protected from contact with the silk and apply it to phytoseiid species for which behavioural performance in the web is known from the literature. It is found that the better protected species experience less hinder from complex webs, such as produced byTetranychus spp. Moreover, they appear to be more successful in exterminating local populations ofTetranychus spp. Thus, setal patterns in the Phytoseiidae may well be a good indicator for selecting candidates for biological control ofTetranychus spp. Finally, we propose a parsimonious scenario for the evolution of dorsal chaetotaxy in the Phytoseiidae.     

Root turnover as determinant of the cycling of C,N, and P in a dry heathland ecosystem

Root production and turnover were studied using sequential core sampling and observations in permanent minirhizotrons in the field in three dry heathland stands dominated by the evergreen dwarfshrub Calluna vulgaris and the grasses Deschampsia flexuosa and Molinia caerulea, respectively. Root biomass production, estimated by core sampling, amounted to 160 (Calluna), 180 (Deschampsia) and 1380 (Molinia) g m^-2 yr^-1, respectively. Root biomass turnover rate in Calluna (0.64 yr^-1) was lower compared with the grasses (Deschampsia: 0.96 yr^-1; Molinia 1.68yr^-1)). Root length turnover rate was 0.75–0.77 yr^-1 (Deschampsia) and 1.17–1.49 yr^-1 (Molinia), respectively. No resorption of N and P from senescing roots was observed in either species. Input of organic N into the soil due to root turnover, estimated using the core sampling data, amounted to 1.8 g N m^-2 yr^-1(^Calluna), 1.7 g N m^-2 yr^-1 (Deschampsia) and 19.7 g N m^-2 yr^-1 (Molinia), respectively. The organic P input was 0.05, 0.07 and 0.55 g P M^-2 yr^-1, respectively. Using the minirhizotron turnover estimates these values were20–22% (Deschampsia) and 11–30% (Molinia) lower.When the biomass turnover data were used, it appeared that in the Molinia stand root turnover contributed 67% to total litter production, 87% to total litter nitrogen loss and 84% to total litter phosphorus loss. For Calluna and Deschampsia these percentages were about three and two times lower, respectively.This study shows that (1) Root turnover is a key factor in ecosystem C, N, and P cycling; and that (2) The relative importance of root turnover differs between species.     

The carbohydrases of Bacillus stearothermophilus NCIB 11412 and NCIB 10814

Summary Two strains (NCIB 11412 and NCIB 10814) of the thermophilic organism Bacillus stearothermophilus were found to produce complex carbohydrase systems. The enzyme activities in each system include -amylase as the major component, maltase, pullulanase, a minor amylase and cyclodextrinase. The latter three activities are produced in low yield in both strains. A crude enzyme preparation from each strain possessed maltogenic properties on hydrolysis of soluble starch. Following rigorous purification procedures, the purified major -amylase from either strain did not produce maltose as a major end-product of starch hydrolysis. However, a partially purified mixture of pullulanase, minor amylase and cyclodextrinase activities from NCIB 11412 and NCIB 10814 produced 56.4% and 62.0% maltose, respectively, from soluble starch.     

Germination and early establishment of lower salt-marsh species in grazed and mown salt marsh

Abstract. Lower salt-marsh species occur more in the grazed than in the mown sites of a salt marsh in Schiermonnikoog in the Netherlands. This was demonstrated by a sowing experiment which determined which characteristic of the stand structure, height of the canopy or percentage of bare soil, is responsible for this difference. The total number of seedlings which emerged was negatively related to the height of the canopy and positively to the percentage of bare soil. The survival of seedlings of Suaeda maritima and Plantago maritima could be explained by the height of the canopy and for the latter species also by the percentage of bare soil. The survival of Aster tripolium and Plantago maritima was higher in grazed than in mown sites. Since the amount of bare soil was higher than seemed necessary for germination and survival, it was concluded that the taller canopy was responsible for the absence of lower salt-marsh species in the mown sites.     

Purification and Properties of Deoxyribonucleic Acid Binding Factor Isolated from the Surface of Streptococcus sanguis Cells

Deoxyribonucleic acid (DNA) binding factor (BF) was found in surface fluids from competent and noncompetent cells of Streptococcus sanguis strains Challis, Wicky, and Blackburn. Fluids from noncompetent cells exhibited about 10% BF activity compared with extracts from competent cells. BF from competent Wicky cells was purified to homogeneity by electrophoresis and immunodiffusion. Purified BF preparations exhibited slight endonucleolytic activity, directed mainly against single-stranded DNA. Nucleolytic and DNA binding activities present in purified BF could be separated by polyacrylamide gel electrophoresis. Purified BF was sensitive to proteolytic enzymes and to phospholipase D, and its activity was stimulated in the presence of low Triton X-100 concentrations. The protein component of BF is a single, monomeric polypeptide with a molecular weight of 56,000 and an isoelectric point of pH 5.8. Binding of purified BF to DNA was a very rapid process at the optimum temperature, pH, and ionic strength and led to the formation of fast-sedimenting complexes. Purified BF was tested for several properties. It exhibited higher affinity to single- than to double-stranded DNA. It bound poorly to glucosylated phage T4 and single-stranded, synthetic polydeoxyribonucleotides and did not bind to RNA. It protected single-stranded DNA against nuclease S₁ action but did not protect native DNA against deoxyribonuclease I action. No evidence was found for unwinding activity, using double-stranded DNA as a substrate.     

Phytoplankton/zooplankton relationships during spring in the brackish Lake Grevelingen 1976–1978 (III)

Summary Rotifer species of the genusSynchaeta represent the dominating zooplankton organisms during early spring (BAKKERet al., 1977).Reproductive capacity of the rotifers (sensu EDMONDSON, 1965) was always significantly correlated with algal densities, cryptomonad flagellates being the main food. The slope of the curve representing the feeding relationSynchaeta-Cryptomonas was significantly steeper than forSynchaeta and the diatomSceletonema. Positive influences of increasing temperature forSynchaeta cf.vorax were only found within the range of 2–7°C. When temperature rose above 10°CS. vorax was succeeded byS.triophthalma. Negative temperatures, occurring in brackish lakes during periods of strong frost, caused a decrease of the egg ratio (numbers of eggs per animal) and of the population density.Development times ofS. vorax eggs were determined (VAN DAMet al., 1978) in order to calculate birth rates and production. Egg development time amounted to 3 days at 5°C and to 2 days at 10°C. Birth rates declined strongly when maximum population densities were reached. Mortality was approached by subtracting the actual rate of increase of population density from the birth rate. Sometimes negative mortalities were found, presumably caused by the development of resting eggs.Population dynamics of the rotifers were related to phytoplankton biomass and to primary productivity. Grazing effects of the rotifers on the algal population became evident during the final stage of the spring bloom (April). In the beginning (February–March) the mortality of the algal population could not be explained by grazing, other factors (cf. JASSBY and GOLDMAN, 1974) being responsible.