Fluorescence-microscopical demonstration of a population of gastro-intestinal nerve fibres with a selective affinity for Quinacrine

Summary A population of nerve fibres in the gastro-intestinal tract of mice showing a high affinity for quinacrine was revealed by fluorescence microscopy. Similar results were obtained in rats and guinea pigs. Whole-mounts of sheets of the smooth muscle layer following incubation in 10^-6-10^-7 M quinacrine for 15–60 min revealed fine fluorescent varicose nerve fibers in the myenteric plexus of Auerbach both around nerve cell bodies and in the interconnecting strands. Many fibers were also present between the strands of the plexus, especially running parallel to the circular muscle layer. Such fibers were not seen in similarly quinacrine-incubated irides. A proportion of the cell bodies in Auerbach's plexus also showed quinacrine accumulation. These cells were apparently smaller neurons, sometimes with fluorescent processes. Intraperitoneal injections of quinacrine failed to demonstrate nerve fibers, but some cell bodies in Auerbach's plexus were positive. Subsequent paraformaldehyde treatment for monoamine visualization showed persistent adrenergic nerve terminals in the intestine and iris. These nerves seemed to be fewer and had a more yellow fluorescence than normally. The identity of the quinacrine-positive fibers is discussed with respect to recent suggestions that purinergic, substance P, enkephalin, and somatosin-containing nerves, in addition to adrenergic and cholinergic nerves, are present in the gut wall.Supported by the Swedish Medical Research Council (04X-03185). Magnus Bergvalls Stiftelse and Karolinska Institutets Fonder. For generous gifts of Mepacrine we thank Winthrop, Skärholmen, Stockholm, Sweden. The skilful technical assistance of Miss Gerd Boetius and Miss Maud Eriksson is gratefully acknowledged     

RNA POLYMERASE ACTIVITY IN VARIOUS CLASSES OF NUCLEI FROM DIFFERENT REGIONS OF RAT BRAIN DURING POSTNATAL DEVELOPMENT

—The changes in the wet weight and the numbers of cell nuclei recovered from the cerebral hemispheres, the cerebellum and the brain stem of rats from the period of 5–30 days after birth have been determined. In parallel a study has been made of the RNA polymerase activity, both in the unfractionated nuclei from these regions and in the nuclei separated by zonal centrifugation. In general there is a considerable decline in activity during this period, which occurs in all class of nuclei although not to the same extent. The most dense nuclei from the cerebellum retain relatively high activity at 20 days after birth, possibly due to the contribution of the microneuronal nuclei.     

The effect of convulsions induced by flurothyl on ribonucleic acid synthesis in rat cerebral cortex during the recovery phase.

The effect of convulsions, induced by flurothyl, on RNA synthesis in purified unfractionated nuclei and the cytoplasm of rat cerebral cortex was studied by using a double-label technique involving injection of [3H]- and [14C]-orotate intracisternally. 2. Intact RNA was extracted in 80% yield by an enzymic method by using a proteinase in the presence of sodium dodecyl sulphate followed by deoxyribonuclease. Electrophoresis on 1.5% polyacrylamide-0.5% agarose gels revealed the presence of giant nuclear RNA of size up to approx. 300X 10(6) daltons and mRNA of maximal mol.wt. 9 X 10(6)-16 X 10(6). 3. Nuclear RNA synthesis was decreased to 27% in the first 15 min after convulsions but rapidly increased, so that at 1 1/2 h it was 124% of the control, and at 6 h 147%. 4. Labelling of cytoplasmic RNA was decreased to 15% at 15 min after convulsions but had not recovered to control values by 6 h. 5. Analysis of radioactive gel patterns and the 3H/14C ratio at six time-points (15 min-6h) showed that the major effect was inhibition of the processing of heterogeneous nuclear RNA resulting in a sharp decline in the export of newly synthesized RNA from the nucleus. 6. Cytoplasmic RNA patterns indicated that specific messengers were synthesized at different times during the recovery of the cell after convulsions.     

Effect of tortuous extracellular pathways on resistance measurements

There are many instances in which we are limited to measuring macroscopic quantities such as a bulk flow or an average field. In biology, wer are frequently interested in using such macroscopic measurements, for example, the total current from a tissue, to determine the microscopic properties of the cells or tubules of the tissue. The microstructure of the tissue will generally increase the resistance to flow over what would be measured in an unstructured medium. This paper derives a fairly general expression for the relationship between effective resistance to macroscopic flow and the specific resistance of the medium conducting the microscopic flow. This expression, called a tortuosity factor, is defined entirely in terms of measurable morphometric and geometric parameters of the tissue.     

Toward a second generation of fluorocarbon emulsions. Methodologies: synthesis of fluorocarbons, formulation, preparation and control of emulsions

The fluorocarbons to be used as intravascular gas carriers may be prepared: by fluorination of hydrocarbons, by means of gaseous fluorine or of metallic fluorides or by electrolysis in anhydrous hydrogen fluoride; by selective synthesis from small fluorinated molecules. Selective synthesis seems preferable because of the high purity of the compounds thus obtained, allowing fairly simple purification and detoxification procedures. The adequacy of fluorocarbons for intravascular use (limited by the values of some of their crucial physicochemical properties such as vapour pressure, gas dissolving power, viscosity) was found dependent of their molecular weight range, preferably between 460 and 540 dalton. Ionic balance, osmolarity, viscosity, mean particle size and particle size distribution are among the most important criteria to be respected in the preparation of biocompatible emulsions. Several physicochemical methods were developed for selecting the best ingredients and the best emulsification procedures and for evaluating the stability of the emulsions to storage and to thermal stress. The combination of these physicochemical measurements may be used as a methodology for the optimisation and control of fluorocarbon emulsions as well as a way of evaluating new ingredients and formulations.