A study of the oxidation of butan-1-ol and propan-2-ol by nicotinamide-adenine dinucleotide catalysed by yeast alcohol dehydrogenase.

1. The kinetics of oxidation of butan-1-ol and propan-2-ol by NAD+, catalysed by yeast alcohol dehydrogenase, were studied at 25 degrees C from pH 5.5 to 10, and at pH 7.05 from 14 degrees to 44 degrees C, 2. Under all conditions studied the results are consistent with a mechanism whereby some dissociation of coenzyme from the active enzyme-NAD+-alcohol ternary complexes occurs, and the mechanism is therefore not strictly compulsory order. 3. A primary 2H isotopic effect on the maximum rates of oxidation of [1-2H2]butan-1-ol and [2H7]propan-2-ol was found at 25 degrees C over the pH range 5.5-10. Further, in stopped-flow experiments at pH 7.05 and 25 degrees C, there was no transient formation of NADH in the oxidation of butan-1-ol and propan-2-ol. The principal rate-limiting step in the oxidation of dependence on pH of the maximum rates of oxidation of butan-1-ol and propan-2-ol is consisten with the possibility that histidine and cysteine residues may affect or control catalysis.     

DEVELOPMENT OF THE EPIPHYLLOUS INFLORESCENCE OF HELWINGIA JAPONICA (HELWINGIACEAE)

The inflorescence of Helwingia japonica (Thunb.) Dietr. is initiated adjacent to the leaf axil on the adaxial side of the base of a leaf primordium during its second plastochron. The inflorescence which develops from the resulting primordium comes to be situated on the midrib of the mature fertile leaf, through the action of a basal, intercalary meristem. In fertile leaves this meristem develops beneath, as well as above, the insertion of the inflorescence primordium on the leaf primordium. The same meristem is present in sterile leaves as well. A separate, adaxial vascular bundle departs from the leaf trace in the base of the petiole and leads to the inflorescence, in the mature fertile leaf. This adaxial vascular bundle is absent in sterile leaves. It is argued that the vascular anatomy does not conclusively confirm the hypothesis that the epiphyllous inflorescence is the congenital fusion product of a leaf and an axillary inflorescence. Instead, it is suggested that the interplay of changes in the position of primordium initiation, and intercalary growth, offers an ontogenetic explanation of the situation, which in turn may be related to the phylogeny of the species in question. It appears to be misguided and futile to look for homologies (i.e., 1:1 correspondences) between fertile and sterile leaves, since 1:1 correspondences do not exist in this case.     

Heterologously expressed polypeptide from the yeast meiotic gene HOP1 binds preferentially to yeast DNA.

HOP1 protein, present in sporulating cells of Saccharomyces cerevisiae and believed to be a component of the synaptonemal complex, has been expressed in Escherichia coli fused to a biotinylated tag protein. Once solubilized from bacterial inclusion bodies, the HOP1 fusion protein was purified by using a combination of avidin-affinity chromatography and gel filtration FPLC and refolded. Sequence comparisons indicate that the HOP1 gene product contains a zinc finger motif, which may confer DNA binding properties, and the recombinant polypeptide was used to assess the putative DNA binding properties of the product of native HOP1 protein using a gel-shift assay. Protein and protein-DNA complexes were detected by exploiting the affinity of streptavidin-alkaline phosphatase for the biotinylated tag protein after Western blotting. The HOP1 fusion protein bound unambiguously to digested genomic yeast DNA. This binding possessed some degree of specificity, was maintained under a wide range of salt concentrations, and was unaffected by the presence of high concentrations of competitor DNA (synthetic poly[dI-dC].poly[dI-dC]). In contrast, no shift was detected when the fusion protein was incubated with digested genomic DNA from Arabidopsis, or with lambda/HindIII DNA. Incubation with digested genomic DNA from Lilium produced a small change in the mobility of the protein. The biotinylated tag protein failed to show any DNA binding activity. Scatchard analysis indicated an apparent yeast genomic DNA:HOP1 fusion protein dissociation constant of K(d) = 5 x 10(-7) M.     

FEMORAL NECK FRACTURES IN THE AGED

Clinical data were reviewed on 99 patients with 102 fractures of the femoral neck on the Orthopedic Service of the San Diego County General Hospital. All were elderly. Twenty-one were dead within two month of the time the fracture was reduced and 36 within a year. In 14 cases the fracture was impacted or undisplaced; there was only one of these in which the fracture did not unite following treatment, and that was for iatrogenic reasons. Sixty-one per cent of the displaced fractures healed; only 32 per cent of the total number of patients with displaced femoral neck fractures walked again.     

Sensitive fluorescence in situ hybridization signal detection in maize using directly labeled probes produced by high concentration DNA polymerase nick translation

The signal produced by fluorescence in situ hybridization (FISH) often is inconsistent among cells and sensitivity is low. Small DNA targets on the chromatin are difficult to detect. We report here an improved nick translation procedure for Texas red and Alexa Fluor 488 direct labeling of FISH probes. Brighter probes can be obtained by adding excess DNA polymerase I. Using such probes, a 30?kb yeast transgene, and the rp1, rp3 and zein multigene clusters were clearly detected.