Ethanol production by anaerobic thermophilic bacteria: regulation of lactate dehydrogenase activity in Clostridium thermohydrosulfuricum

Summary The enzyme lactate dehydrogenase (LDH) in Clostridium thermohydrosulfuricum is controlled by the type and the concentration of the substrate. In batch fermentations an increase of the initial concentration of glucose leads to an increase in the activity of LDH. This increase in activity is related to the accumulation of fructose 1,6-diphosphate (F 1,6-DP), an intermediate of the Embden-Meyerhof-Parnas (EMP) pathway, which stimulates the enzyme by increasing its affinity for pyruvate and NADH. The K _mvalues of LDH for pyruvate and NADH, which are 2.5×10^-3 M and 9.1×10^-5 M respectively in absence of F 1,6-DP, fall considerably in the presence of this substrate. In presence of 0.2 mM of F 1,6-DP we observed a K _mof 3.3×10^-4 M for pyruvate and 4.1×10^-5 M for NADH.     

Primary attraction and random landing in host‐selection by wood‐feeding insects: a matter of scale?

1 Most plant‐feeding insects show some degree of specialization and use a variety of cues to locate their host. Two main mechanisms of host location, primary attraction and random landing, have been investigated for such insects. 2 Research has led to contradictory conclusions about those hypotheses, especially for wood‐feeding insects; however, recent studies suggest that both mechanisms may take place in a single taxon but at different scales. 3 We developed a field experiment to test the hypothesis that primary attraction occurs at larger scale and random landing at finer scale in wood‐feeding insects. Landing rates, measured using sticky traps, were compared first between patches and then between individual trees according to their distance to a baited central tree. 4 Polynomial functions describing landing rate to distance relationships were compared with a function produced by a null model describing what should occur under the random landing hypothesis. Scolytidae and Cerambycidae (Coleoptera) responded to volatiles at the patch scale, supporting the primary attraction hypothesis, but the landing patterns of some groups at finer scale matched closely the predictions of our null model, giving support to the random landing hypothesis. 5 Our results show that the primary attraction and random landing hypotheses are not mutually exclusive and that prelanding use of host‐produced volatile is scale‐dependant. Scale considerations should thus be included in the study of prelanding host‐selection of wood‐feeding insects.     

Use of molecular techniques for analysis of Ia structure-function relationships

An overview of a strategy for the molecular analysis of class II major histocompatibility complex (Ia) gene product structure-function relationships is presented, and results obtained to date by using this approach are summarized. The A beta, A alpha, E alpha, and E beta genes have been cloned and sequenced to yield information on gene organization and primary protein sequence. Comparison of sequences from allelic forms of these genes show the NH2-terminal domain to be the locus of most intraspecies polymorphism. Transfection of I-A alpha and A beta genes into B lymphoma cells or L cells has generated cells expressing the transfected gene products on their membrane. Such Ia+ transfectants present antigen to various T cells, which use the expressed I-A as a restriction element. Exon shuffling has shown the beta 1 domain of A beta to play a predominant role in such restricted antigen recognition. Preliminary data refining this analysis to sites within beta 1, as well as data on control of alpha: beta chain association, are reviewed, and future prospects for use of this approach in resolving questions of immunological interest are discussed.     

Location of respiration activity in filamentous bacteria by image analysis

Respiration activity was located in Streptomyces ambofaciens using formazan crystals and image analysis. The technique was validated by comparing it to a global respiration rate measurement method. A uniform distribution of formazan crystals along the hyphae was found in cultivations in flasks and fermenter, throughout the cultures, although respiration is globally decreased when cultures age.     

Fluorescence-assisted mismatch analysis (FAMA) for exhaustive screening of the α-galactosidase A gene and detection of carriers in Fabry disease

We used the fluorescence-assisted mismatch analysis (FAMA) method to screen rapidly the α-galactosidase A gene in patients with Fabry disease in order to identify unknown mutations and help define genotype-phenotype correlations in this X-linked lysosomal storage disorder. Chemical cleavage at mismatches on heteroduplex DNA end-labeled with strand-specific fluorescent dyes, reliably detects sequence changes in DNA fragments of up to 1.5 kb and locates them precisely. Exhaustive scanning of the α-galactosidase gene was accomplished on four polymerase chain reaction-generated amplicons, covering all seven exons, the exon-intron boundaries, and 700 bp of 5′-flanking sequence. Mutations were identified in each of the 15 patients studied from nine unrelated kindreds. Among the seven previously undescribed sequence changes, three are obviously pathogenic because they lead to premature protein termination. The other four, a splice-site mutation and three missense mutations, were the only changes found upon complete scanning of the gene and its promoter. In addition, FAMA also detects female heterozygous carriers more dependably than direct sequencing, and thus provides a valuable diagnostic test. In Fabry disease, this molecular criterion is especially important for genetic counseling since heterozygotes can be asymptomatic and their enzymatic values within the normal range. Received: 9 April 1996 / Revised: 8 July 1996     

Quantitative characterization of cellular differentiation of Streptomyces ambofaciensin submerged culture by image analysis

Image analysis methods were developed for light and epifluorescence microscopy of Streptomyces ambofaciens undergoing differentiation in submerged culture. Grey level images were obtained with an integration controlled CCD camera and allowed three parameters to be measured: occurence of empty zones in mycelium, number of septations, mycelium thickness.