Mevalonate activating enzymes and phosphatases in higher plants

The pH optima of mevalonate kinase and phosphatases in green leaves, cotyledons and chloroplasts of French bean, and in green leaves and chloroplasts of maize, have been studied. Whereas in chloroplasts the pH optimum for mevalonate kinase is at pH 7·5 with little or no activity at pH 5·5, there is with leaf and cotyledon preparations appreciable activity at the lower pH. Under some circumstances isoelectric focusing studies have given fractions showing mevalonate kinase activity at only pH 7·5 or 5·5. Acid phosphatase and ATPase activity in preparations is maximal at pH 5·5 and is much reduced in the presence of high levels of phosphate. Other investigations reported concern the stability of mevalonate kinase and phosphatase activity at pH 5·5 and 7·5 on ageing of extracts, and the activity of mevalonate kinase on greening of etiolated French bean cotyledons. The influence of metal cofactors and fluoride on mevalonate kinase and phosphatase are reported.     

Trypanosoma congolense: mechanical removal of the surface coat in vitro

By shaking suspensions of Trypanosoma congolense in isotonic buffer the surface coat could be separated from the cell body. The release of radioactivity from trypanosomes, selectively labeled in the surface coat by diazoniobenzenesulfonate, was used to follow the kinetics of coat detachment. The proteins in the supernatants of shaken trypanosomes were analyzed by sodium dodecyl sulfate—polyacrylamide gel electrophoresis. The shaking conditions had to be carefully controlled to avoid complete rupture of trypanosomes. Otherwise the coat protein was rapidly degraded by endogenous proteases. The influence of several parameters on the yield of coat release and the degree of degradation of the coat protein was investigated, including the ratio of trypanosome suspension volume to shaking vessel volume, vessel surface, temperature, shaking frequency, and preincubation of the trypanosomes at 0 C. By combining these parameters an optimal scheme was developed which allowed the separation of more than 90% of the coat protein from T. congolense, the detached protein showing no degradation at all. These results could be confirmed by electron microscopy of shaken and unshaken trypanosomes.     

Evaluation of genetic risks of alkylating agents IV. Quantitative determination of alkylated amino acids in haemoglobin as a measure of the dose after-treatment of mice with methyl methanesulfonate.

The present study explores the possibilities of using specific amino acids in haemoglobin for tissue dosimetry of alkylating agents. The well-known directly alkylating compound methyl methanesulfonate has been used as a model compound.In one experiment ³H-labelled methyl methanesulfonate was given to mice intraperitoneally at three dose levels. The degree of alkylation of haemoglobin exhibited a linear dependence on the quantity of methyl methanesulfonate injected. The degree of alkylation of guanine-N-7 in DNA indicated a slight positive deviation from linearity at high doses.After a single injection the degree of alkylation of cysteine-S and histidine-N-3 in haemoglobin decreased linearly with time reaching the value zero after about 40 days (the life-time of the erythrocytes in the mouse). This demonstrates a stability of these alkylated products, which is fundamental to their use as integral dose monitors.In a second experiment mice were treated with methyl methanesulfonate once a week over a period of 8 weeks. The experiment demonstrated an accumulation of alkylated groups in haemoglobin in agreement with expectation.A method for the quantitative determination of S-methylcysteine in a protein hydrolysate by gas chromatography was developed.     

Leishmania mexicana: inhibition and stimulation of phagosome-lysosome fusion in infected macrophages

The polyanionic compound poly-d-glutamic acid was found to inhibit significantly the fusion of secondary lysosomes to phagosomes containing Leishmania mexicana mexicana amastigotes for at least 96 hr. This process was viewed both by dark-field vital fluorescence microscopy and transmission electron microscopy. In poly-d-glutamic acid-treated macrophages parasites multiplied at a significantly greater rate than in untreated macrophages. Conversely, the secondary amine chloroquine caused a marked reduction in parasite growth. When L. m. mexicana promastigotes were substituted for amastigotes these results were strikingly more pronounced.     

Simple and fast purification of Escherichia coli adenylate kinase

Adenylate kinase from E. coli (strains CR341 and CR341 T28, a temperature-sensitive mutant) was purified by a two-step chromatographic procedure. The enzyme from crude extracts of both mutant and parent strain was bound to blue-Sepharose at pH 7.5, thereafter specifically eluted with 0.05 mM P1,P5-di(adenosine-5')pentaphosphate. A second chromatography on Sephadex G-100 yielded pure enzyme. E. coli adenylate kinase was strongly inhibited by P1,P5-di(adenosine-5')pentaphosphate (Ki 0.6 microM for adenylate kinase of strain CR341 and 2.1 microM in the case of mutant enzyme). After denaturation in 6 M guanidinium hydrochloride both mutant and parent adenylate kinase returned rapidly to the native, active state by dilution of guanidinium hydrochloride.     

The influence of spatial scale on quantifying insect dispersal: an analysis of butterfly data

Abstract. 1. A linear relationship between mean movement distances recorded and the size of the study area was found in a comparison of five mark–release–recapture studies of the meadow brown butterfly.
2. The scale impact on mean butterfly movement distances was also apparent when comparing the results for different butterfly species reported in 21 mark–release–recapture studies.     

Synchrony in population counts predicts butterfly movement frequencies

1. Measuring functional connectivity, the ability of species to move between resource patches, is essential for conservation in fragmented landscapes. However, current methods are highly time consuming and expensive. 2. Population synchrony‐ the correlation in time series of counts between two long‐term population monitoring sites, has been suggested as a possible proxy measure of functional connectivity. To date, population synchrony has been shown to correlate with proxies for movement frequency such as the coverage of suitable habitat types in intervening landscapes, and also least cost distances around hostile land cover types. 3. This provides tentative evidence that synchrony is directly driven by movements of the focal species, but an alternative explanation is that these land cover types affect the movement of interacting species (e.g. natural enemies of the focal species) which can also drive synchronous population dynamics. Therefore, what is needed is a test directly relating population synchrony to movement frequencies of a focal species. 4. Here we use data from a 21 year mark‐release‐recapture study and show that population synchrony does indeed predict movements of a focal butterfly species Boloria eunomia (Esper). 5. There is growing evidence that population synchrony can be a useful conservation tool to measure functional connectivity.