Determination of products of acetohydroxy acid synthase by the colorimetric method, revisited

The enzyme acetohydroxy acid synthase (AHAS, EC 4.1.3.18) catalyzes two competing reactions of physiological importance: condensation of two molecules of pyruvate to form acetolactate (AL) or condensation of pyruvate and 2-ketobutyrate to form acetohydroxybutyrate (AHB). The activity of AHAS is most frequently analyzed using the Westerfeld method, in which the acetoin formed upon decarboxylation of AL is determined by colorimetric reaction with creatine and alpha-naphthol. However, there has been confusion as to the interpretation of the results of this assay in the presence of both substrates, conditions which lead to formation of both AL and AHB. By applying this assay to enzymatically prepared samples of AL and AHB which have also been analyzed by two other independent methods, we show here that the color yield for AHB in the commonly used assay is 35-40% that for equivalent amounts of acetoin or AL. The relative color yield is not significantly affected by varying the time or temperature of various steps in the color-forming reaction. This information could in principle be used, together with an independent specific assay for AHB, to determine the composition of an AHAS product mixture; it would, however, be less accurate than a simultaneous chromatographic method.     

A comparison of vector population indices for forecasting barley yellow dwarf virus in autumn sown cereal crops

Two approaches based on the concept of a vector population index are considered as possible deterministic elements for an empirical forecast of barley yellow dwarf virus (BYDV) in autumn sown cereals. The first, an aerial vector index, is a further elaboration of the infectivity index proposed by Plumb, Lennon & Gutteridge (1981), which assumes that virus damage is a function of the number of infective migrant alatae of the two main aphid vectors, Rhopalosiphum padi L. and Sitobion avenae F., integrated over time from crop planting or emergence. The new formulation, however, excludes holocyclic alate morphs (i.e. males and gynoparae) of the former species, which, although generally abundant in autumn, are nevertheless perceived as relatively unimportant virus vectors since they colonise only the alternative woody host, Prunus padus (the bird-cherry tree). The second approach, a crop vector index, is a more fundamental departure which argues that field populations of viruliferous aphids, both alatae and apterae, which have already colonised cereals, may be a better criterion of potential virus spread than the density of aerial migrant vectors. This index retains a similar integral form, but evaluates crop exposure to BYDV as accumulated infectious aphid-days. A method is described whereby this function can be derived from irregular or infrequent aphid samples in the crop. Both methods, unlike Plumb's (1976) original concept, produced indices which were significantly related to subsequent virus infection and yield loss in winter barley at Long Ashton (S.W. England, UK), 1978–1986. The best models were obtained with the crop vector index, fitted to observed virus infection by generalised linear regression using a complementary log-log link function, or to observed yield loss by simple linear regression using a log transformation of yield (r = 0.84 in each case; compared with r-values > 0.65 for the aerial vector index, and > 0.35 for Plumb's (1976) index). However, the residual errors and hence confidence limits of these fitted regressions were too large for predicting damage that was significantly less than a reasonable economic damage threshold for BYDV control. Analyses of the separate components of each index showed a good general relationship between aphid infectivity and the severity of crop infection, confirming the epidemiological importance of this factor. The functional expressions of aphid density, however, were not significant. This evident weakness in the models, and alternative approaches to BYDV forecasting are discussed.     

Effect of simultaneous application of heat and pressure on the survival of bacterial spores

C.G. MALLIDIS AND D. DRIZOU. 1991. The effect of simultaneous application of moderate hydrostatic pressure (10–300 atm) and heat on the survival of the Bacillus stearothermophilus spores in a flow-through system was investigated. A high heterogeneity of the sensitization of spores to heat by pressure was found. A higher degree of reduction of heat resistance was observed at the low than at the high temperatures tested. The simultaneous application of moderate pressure and heat can not be applied for the preservation of liquid foods due to the extreme heterogeneity of spore sensitization to heat by pressure.     

Chromosome studies in the red howler monkey, Alouatta seniculus stramineus (Platyrrhini, Primates): description of an X1X2Y1Y2/X1X1X2X2 sex-chromosome system and karyological comparisons with other subspecies

In the red howler monkey, Alouatta seniculus stramineus (2n = 47, 48, or 49), variations in diploid chromosome number are due to different numbers of microchromosomes. Males exhibit a Y;autosome translocation involving the short arm of an individual biarmed autosome. Consequently, the sex-chromosome constitution in the male is X1X2Y1Y2, with X1 representing the original X chromosome, X2 the biarmed autosome (No. 7), Y1 the Y;7p translocation product, and Y2 the acrocentric homolog of 7q. In the first meiotic division, a quadrivalent with a chain configuration can be observed in spermatocytes. Females have an X1X1X2X2 sex-chromosome constitution. Chromosome heteromorphisms were observed in pair 13, due to a pericentric inversion, and pair 19, due to the presence of constitutive heterochromatin. Microchromosomes, which varied in number between individuals, were also heterochromatic. NOR-staining was observed at two separate sites on a single chromosome pair (No. 10). A comparison of A.s. stramineus with A.s. macconnelli shows that these two subspecies have identical diploid chromosome numbers (47, 48, or 49), again due to a varying number of microchromosomes, and that they share a similar sex-chromosome constitution. Their karyotypes, however, are not identical, but can be derived from each other by a reciprocal translocation. Further comparisons with other A. seniculus subspecies reported in the literature indicate that this taxon is not karyologically uniform and that substantial chromosome shuffling has occurred between populations that have been considered to be subspecies by taxonomic criteria based on their morphometric attributes.     

Assignment of a human serum glycoprotein SP-40,40 gene (CLI) to chromosome 8.

SP-40,40 is a serum glycoprotein consisting of two different subunits (alpha and beta) assembled into a dimer by disulfide bonds. Northern blot hybridization, using total RNA from several cell lines, showed that SP-40,40 is expressed in glioblastoma and testicular tumor cells, as well as hepatoma cells. Spot blot hybridization of flow-sorted human chromosomes, using a SP-40,40 cDNA fragment as a probe, localized the gene for SP-40,40 to human chromosome 8. This gene has been given the designation CLI, for complement lysis inhibitor, by the Human Gene Nomenclature Committee.