Synthesis of phage-specific transfer RNA molecules by vibriophage phi 149

32P-Labelled tRNA was isolated from uninfected and phage phi 149-infected Vibrio cholerae cells. These tRNA preparations were then hybridised with DNA isolated from phage phi 149. Significant hybridisation was observed only with tRNA from phage phi 149-infected cells. This strongly suggests that infection of classical vibrio with phage phi 149 results in the synthesis of phage-specific tRNA molecules.     

Reaction of organocuprate reagents with protected 1,2-anhydro sugars. Stereocontrolled synthesis of 2-deoxy-C-glycosyl compounds

The reaction of protected 1,2-anhydro-α-d-gluco- and β-d-manno-pyranoses with alkyl and phenyl organocuprates afforded the corresponding C-glycosyl compounds in acceptable to high yield. Complete stereocontrol was obtained, leading respectively to the β-d or the α-d anomer. With the perbenzylated manno derivative, deoxygenation at C-2 was achieved in high yield, affording 2-deoxy-α-d-C-glycosyl compounds.     

Target size analysis by radiation inactivation: a large capacity tube rack for irradiation in a Gammacell 220

Target size analysis by radiation inactivation is now a well-established method to study structure-function relationships in biologically active macromolecules without prior purification or even solubilization. Recently, it was reported that a relatively low-dose-rate but commonly available gamma source such as the Gammacell 220 (Atomic Energy of Canada, Ltd.) can be used to carry out radiation inactivation experiments providing it is appropriately calibrated with enzymes of known radiation sensitivities (G. Beauregard and M. Potier (1982) Anal. Biochem. 122, 379-384). In this report, a tube rack designed to fit into the irradiation chamber of the Gammacell 220 which allows five experiments (at 30 tubes per experiment) to be carried out simultaneously with both standard and unknown samples is described. The dose rates delivered at different positions in the rack were determined by irradiating rat liver cytosolic neuraminidase, an enzyme of known radiation sensitivity. A better than 2.7% agreement was obtained between experimental dose rate and computed values from isodose curves previously published by other authors (O. A. Curzio and H. O. Quaranta (1982) Int. J. Appl. Radiat. Isot. 33, 1-3).     

Imipramine associations with plasma components and its uptake by cultured human cells

It has been proposed that in vivo variability in response to certain hydrophobic chemicals or drugs, such as imipramine, may be due in part to the varying plasma lipid levels in patients. The distribution of [3H]imipramine into the lipoproteins of human plasma was therefore studied. Differential density centrifugation of plasma containing [3H]imipramine resulted in flotation of very low density, low density and high density lipoproteins (VLDL, LDL, HDL) and approximately one-third of the total 3H radioactivity. Twelve percent of the radioactivity was present in the sedimented fraction which included most of the plasma proteins. There appeared to be little specific binding of [3H]imipramine to VLDL or LDL, as shown by ultracentrifugation, dialysis and column chromatography. [3H]Imipramine was readily incorporated into cultured human fibroblasts;o no differences were observed in cellular uptake whether it was added to the medium in plasma, LDL or HDL. Also, no differences in uptake of [3H]imipramine by LDL-receptor positive and receptor negative cells were noted. These experiments indicate that LDL is not a major vehicle for the transport of this drug and that both the bound and free fractions are available for cellular uptake.     

Teleonomical optimization of a fractal model of the pulmonary arterial bed

Modeling the pulmonary arterial tree (PAT) is considered here as an optimal synthesis problem. Firstly, a class of candidate models is specified: the three-dimensional symmetric dichotomous fractal trees of elastic tubes described by Womersley's equations. Secondly, the parameters are shown to be constrained by interactions of PAT with the rest of the body; these constraints are used to limit the volume of the parametric space to which attention will be directed in the synthesis step. Thirdly, a teleonomical hypothesis is proposed: a naturally selected PAT must have a minimal input impedance under conditions keeping total arterial volume and distensibility as small as possible. This hypothesis is translated in mathematical terms and the resulting cost-function minimized in the limited parametric volume. The optimal model has parameter values and an impedance spectrum corresponding satisfactorily with real data. Moreover this model gives a clear picture of the internal hemodynamic behavior of PAT as an impedance matching device.     

The exchange hypothesis for the formation of chromatid aberrations: an experimental test using bleomycin

The association between bleomycin-induced chromatid aberrations and BUdR-label exchange between sister chromatids was investigated in order to evaluate Revell's exchange hypothesis for the formation of chromatid aberrations. The results of this study indicate that a larger than expected proportion of chromatid breaks can be accounted for by the exchange hypothesis though not all breaks are the result of incomplete exchange.     

Cross-resistance studies with V79 Chinese hamster cells adapted to the mutagenic or clastogenic effect of N-methyl-N′-nitro-N-nitrosoguanidine

When V79 cells are exposed to a single low dose of MNNG or MNU they acquire resistance to the mutagenic or to the clastogenic effect of the agents. Here the effect of MNNG pretreatment on mutagenesis (6-thioguanine resistance) and aberration formation in cells challenged with various mutagens/clastogens is reported. MNNG-adapted cells were resistant to the mutagenic effects of MNU and, to a lower extent, of EMS. No mutagenic adaptation was observed when MNNG-pretreated cells were challenged with MMS, ENU, MMC or UV.

Cells pretreated with a dose of MNNG which makes them resistant to the clastogenic effect of this compound were also resistant to the clastogenic activity of other methylating agents (MNU, MMS), but not so with respect to ethylating agents (EMS, ENU). Cycloheximide abolished the aberration-reducing effect of pretreatment. However, when given before the challenge dose of MNNG, MNU or MMS, it drastically enhanced the aberration frequency in both pretreated and non-pretreated cells. No significant enhancement of aberration frequency by cycloheximide was found for ethylating agents.

The results indicate that clastogenic adaptation is due to inducible cellular functions. It is concluded that mutagenic and clastogenic adaptation are probably caused by different adaptive repair pathways.     

The centrifugal horizontal cells in the lobula plate of the blowfly, Phaenicia sericata

Intracellular recordings combined with iontophoretic injection of Procion Yellow M4RAN were used to study the anatomy and physiology of the centrifugal horizontal cells (CH-cells) in the lobula plate of the blowfly, Phaenicia sericata.Anatomy: The CH-cells comprise a set of two homolateral, giant visual interneurones (DCH, VCH) at the rostral surface of each lobula plate. Their extensive arborizations in the lobula plate possess bulbous swellings (boutons terminaux). The arborization of one cell (DCH) covers the dorsal, and the arborization of the other cell (VCH) the ventral half of the lobula plate. Their axons run jointly with those of the horizontal cells through the chiasma internum and the optic peduncle. Their protocerebral arborization possesses spines; they form a dense network together with the axonal arborization of the horizontal cells, a second type of giant homolateral cell most sensitive to horizontal motion. The protocerebral arborization of the CH-cells gives rise to a cell body fibre which traverses the protocerebrum dorsally to the oesophageal canal. The cell body lies on the contralateral side laterally and slightly dorsally to the oesophageal canal in the frontal cell body layer.Physiology: The CH-cells respond with graded potentials to rotatory movements of their surround. Cells in the right lobula plate respond with excitation (excitatory postsynaptic potentials, membrane depolarization) to clockwise motion (contralateral regressive, ipsilateral progressive), and with inhibition (inhibitory postsynaptic potentials, membrane hyperpolarization) to counterclockwise motion in either or both receptive fields; CH-cells respond to motion presented to the ipsilateral and/or contralateral eye. Cells of the left lobula plate respond correspondingly to the reverse directions of motion. Vertical pattern motion and stationary patterns are ineffective.The heterolateral H1-neurone elicits excitatory postsynaptic potentials in the DCH-cell; these postsynaptic potentials are tightly correlated 1:1 to the preceding H1-action potentíal. The delay between the peak of the action potential and the beginning of the DCH-postsynaptic potential is 1.15 msec, agreeing very well with the value reported previously for the blowfly, Calliphora (Hausen, 1976a). The synaptic input and output connections of the CH-cells are discussed.     

Ligation of highly modified bacteriophage DNA

After digestion by TaqI or nicking by DNAase I, five highly modified bacteriophage DNAs were tested as substrates for T4 DNA ligase. The DNAs used were from phages T4, XP12, PBS1, SP82, and SP15, which contain as a major base either glucosylated 5-hydroxymethylcytosine, 5-methylcytosine, uracil, 5-hydroxymethyluracil, or phosphoglucuronated, glucosylated 5-(4′,5′-dihydroxypentyl)uracil, respectively. The relative ability of cohesive-ended TaqI fragments of these DNAs and of normal, λ DNA to be ligated was as follows: $\text{λ}\text{DNA}\text{=}\text{XP}\text{12}\text{DNA}\text{>}\text{SP}\text{82}\text{DNA}\text{?}\text{nonglucosylated}\text{T}\text{4}\text{DNA}\text{>}\text{T}\text{4}\text{DNA}\text{=}\text{PBS}\text{1}\text{DNA}\text{?}\text{SP}\text{15}\text{DNA}$. TaqI-T4 DNA fragments were also inefficiently ligated by Escherichia coli DNA ligase. However, annealing-independent ligation of DNAase I-nicked T4, PBS1, and λ DNAs was equally efficient. We conclude that the poor ligation of TaqI fragments of T4 and PBS1 DNAs was due to the hydroxymethylation (and glucosylation) of cytosine residues at T4's cohesive ends and the substitution of uracil residues for thymine residues adjacent to PBS1's cohesive ends destabilizing the annealing of the restriction fragments. Only SP15 DNA with its negatively charged, modified base was unable to serve as a substrate for T4 DNA ligase in an annealing-independent reaction; therefore, its modification directly interfered with enzyme binding or catalysis.