Early changes in mitochondrial monoamine oxidase activity of rat liver during 2-acetylaminofluorene feeding

The activities of mitochondrial type A and B monoamine oxidase were determined in the liver of rats fed a diet containing 2-acetylaminofluorene (AAF). Three days after the initiation of AAF-feeding, there was a significant decrease of type B monoamine oxidase activity without affect on type A enzyme. The decreased activity of type B monoamine oxidase, which reached a minimum after three weeks, was sustained for as long as AAF-feeding was continued. Sex-related difference in response to AAF was seen in the rat with respect to the onset and the intensity of the decreased type B monoamine oxidase activity, male rats being more sensitive to the carcinogen than female rats. In contrast to the in vivo effect, AAF showed a potent inhibitory effect on type A monoamine oxidase, rather than on type B enzyme, when added in vitro. The pI₅₀ values were estimated to be 7.5 against type A monoamine oxidase and 4.1 against type B enzyme, respectively. The in vitro inhibition of both types of monoamine oxidase by AAF was competitive. The K_i values for AAF were calculated to be 9.51 · 10^?9 M for type A monoamine oxidase and 1.30 · 10^?5 M for type B enzyme, respectively. In accordance with the potent inhibitory effect of AAF on type A monoamine oxidase in vitro, a single administration of the carcinogen, at a dose of 50 mg/kg, resulted in a marked and temporal decrease of the enzyme activity in the mitochondria of male rat liver. Recovery of the decreased type B monoamine oxidase activity was slow, and the enzyme activity did not return to control levels, even if rats were fed the basal diet for 2 or 4 weeks after the cessation of AAF-feeding.     

Sequential polymerization of flagellin A and flagellin B into Caulobacter flagella

The mode of polymerization of two species of flagellins, flagellin A and flagellin B, in polar flagella of Caulobacter crescentus was examined. By immunological staining we found that 1 to 1.2 μm of the portion of the flagellar filament proximal to the cell was composed of flagellin B, whereas about 5 μm of the distal portion was composed of flagellin A. This result, together with the previous observation that a flagellin B-less mutant cannot form normal flagella but instead forms stubs in spite of their high level of flagellin A synthesis, indicates that flagellin B is very important for the formation of complete flagella and/or for the initiation of filament formation from the hook.     

Automated high-performance liquid chromatographic method for determination of furosemide in dog plasma

An automated high-performance liquid chromatographic method for the determination of the diuretic drug furosemide has been established. Dog plasma was injected directly into a two-column system with a BSA—ODS (ODS column coated with bovine serum albumin) precolumn and a C₁₈ analytical column for the separation of furosemide. The two columns were automatically switched. Furosemide remained trapped on the precolumn while proteins were eluted to waste. After column switching, furosemide was washed onto the analytical column and analysed without interference. The greatest advantage of the method is its easy performance without manual sample preparation; it requires no extraction or deproteinization. The method allows determination of 0.1–10 μg/ml of furosemide with accuracy and precision comparable with previously reported values. The coefficients of variation obtained from replicate measurements of 1 μg/ml and 5 μg/ml samples were 1.65% and 2.40%, respectively. This method was used to measure the plasma levels of furosemide in beagle dogs to whom the drugs was administered, as a reference, in a toxicological study.     

Solution properties of synthetic polypeptides. V. Helix–coil transition in poly(β-benzyl L-aspartate)

Simple approximate expressions have been derived from the theory of Zimm and Bragg for use in the analysis of experimental data on the helix-coil transition in polypeptide. On the basis of the resulting expressions practical procedures are proposed to determine two basic parameters characterizing a thermally induced transition, i.e., helix initiation parameter σ and enthalpy change for helix formation, ΔH. They have been applied to the data for poly(β-benzyl L -aspartate) (PBLA) with the result: σ = 1.6 × 10^?4 and ΔH = ?450 cal/mole for PBLA in m-cresol; σ = 0.6 × 10^?4 and ΔH = 260 cal/mole for PBLA in chloroform containing 5.7 vol-% of dichloroacetic acid. This result gives evidence that σ may change not only from one polypeptide to another but also for a given polypeptide in different solvents. The change in limiting viscosity number [η] accompanying the transition was measured in the same solvents. The curve of [η] versus helical content had a relatively monotonic shape for the chloroformdichloroacetic acid solutions as compared with that for the m-cresol solutions, indicating that [η] depended largely on σ. Provided that [η] is a direct measure of the mean-square radius of gyration, 〈S²〉, the results are consistent with the theoretical predictions of Nagai and of Miller and Flory for 〈S²〉.     

Induced single fertilization in maize

 Bicellular pollen with one vegetative nucleus and one diploid arrested generative cell (”monospermic” pollen) was induced by trifluralin treatment of diploid maize plants at 7–9 days before flowering. The arrested generative cell (seemingly a diploid sperm cell) fused with the central cell of diploid plants and produced shriveled endosperm resembling that of a 2n×4n cross in maize. Dual pollination experiments with a purple embryo marker revealed single fertilization events in which the union of one sperm cell with the egg occurs but there is no union of a second sperm cell with the central cell. Singly fertilized ovules survived at least 4 days. Furthermore, many viable triploid plants were obtained. This technique therefore appears to have the potential for manipulating ploidy level in crops and may become useful in investigating fertilization mechanisms of angiosperms. Received: 1 October 1996 / Revision accepted: 8 January 1997     

Adjuvant actions of polyclonal lymphocyte activators: III. Two distinct types of T-initiating adjuvant action demonstrated under different experimental conditions

We demonstrated that each of various polyclonal lymphocyte activators (PLA) exhibits two types of adjuvant action to initiate the carrier-specific helper T-cell response to otherwise nonimmunogenic antigen. Type 1 action was characterized as that to initiate the T-cell response to subcutaneous injection of soluble bovine γ-globulin (BGG), and type 2 as that to initiate the response to intravenous injection of aggregated BGG. Each of various PLA showed these two types of adjuvant action in a dissociated fashion. The capsular polysaccharide of Klebsiella pneumoniae (CPS-K) showed both types of action to the highest degrees. Lipopolysaccharide of Escherichia coli exhibited type 2 action as markedly as CPS-K, but failed to show type 1 action. Concanavalin A showed definite type 1 action, but not type 2 action. Polyadenylic-uridylic acid showed definite type 2 action, but not type 1 action. Type 1 and type 2 actions of dextran sulfate were minimal. A hypothetical view is presented to consider that type 1 adjuvant action is directed to two mutually independent sites whereas type 2 action is directed to one site.