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Biomedical subjects

R E Kingsley

Publications and source records attributed to R E Kingsley.

9 recordsLinked to original sources

Colchicine treatment in the preparation of varicella-zoster virus inocula.

Two methods, ultrasonic and colchicine treatment, for preparing varicella-zoster virus (VZV) inocula were compared. Ultrasonic treatment was unsuccessful in achieving the release of detectable, infectious, cell-free VZV from infected human foreskin fibroblast (SF) cells. Sonication of infected SF cells resulted in losses of virus infectivity in direct correlation with losses in cell viability (r = 0.82). In the second method of generating VZV inocula, VZV-infected SF cells were treated for 15 min with 15-20 ng of colchicine per cell. Colchicine treatment maintained short-term survival of virus infectivity, but resulted in a dramatic decrease in long-term cell viability. In addition, colchicine-treated cells could not support the replication of VZV.

Cell Survival

Sensory and sympathetic innervation of the mammalian cornea. A retrograde tracing study.

The method of retrograde transport of horseradish peroxidase-wheat germ agglutinin (HRP-WGA) was used to study the locations, numbers and somata diameters of corneal afferent and efferent neurons in four different mammalian species. A 2% solution of HRP-WGA was applied to the central cornea of the rat, rabbit, cat and monkey and the animals were perfusion-fixed 48-96 hr later. HRP-WGA-labeled sensory neurons were distributed relatively uniformly throughout the ophthalmic region of the ipsilateral trigeminal ganglion of the rat, cat and monkey. In marked contrast, labeled cells in the rabbit trigeminal ganglion were clustered in a sharply defined longitudinal column located in the midregion of the ophthalmic area. Occasional cells in some cats and monkeys were observed near, and possibly within, the maxillary region of the ganglion. There was no evidence for a dorsoventral somatotopy of corneal afferent neurons in any species. The majority of the labeled afferent somata were small or medium in size, although some larger diameter neurons were also observed. Modest numbers of labeled neurons were observed in the ipsilateral superior cervical ganglion (SCG) of the rabbit and cat; however, only occasional labeled neurons were observed in the SCG of the rat, and none were seen in the monkey. The labeled SCG cells, when present, were concentrated in the rostral half of the ganglion, although many cells in the cat SCG were also found further caudally. No labeled neurons were found in the middle cervical or stellate ganglia of any animal. The results of this study have revealed the existence of subtle interspecies differences in the organization of the mammalian corneal afferent and efferent innervations.

Animals

Preparation of cultured mammalian cells for transmission and scanning electron microscopy using Aclar film.

Common methods for the preparation of cultured cells for concurrent light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) are not completely satisfactory. This article describes how we grow mammalian cells on plastic disks made from Aclar film. Aclar is a transparent fluorinated-chlorinated thermoplastic that contains no volatile components and is, for all practical purposes, chemically inert. Cells adhere to it readily and remain attached after fixation, dehydration, and critical-point drying or embedding. The film also accepts heavy metal coating by ionic bombardment and is extremely stable in the vacuum of the SEM. LM observations are unhindered by Aclar, since the film is as transparent as glass. Fluorescence microscopy is possible with this film, since it exhibits no detectable autofluorescence. During SEM observation, the film has great dimensional stability, and the cells and heavy metal coating remain attached to the Aclar even under high-resolution operating conditions. TEM processing of specimens grown on Aclar is simplified by the fact that Aclar does not stick to the epoxy resins used in EM. Furthermore, Aclar is easily sectioned and does not damage knives used in ultramicrotomy. The use of Aclar film considerably simplifies the preparation of cultured cells for all types of microscopy. This method is particularly useful in correlating surface features between SEM and TEM observations.

Animals

A proportional controlled thermoregulator circuit of simple design.

This paper describes a proportional controlled thermoregulator circuit. The instrument controls D.C. current in the heating device, precluding the introduction of radiated interference by the heater. This circuit is capable of regulating the temperature of a typical decerebrated cat to within 0.1 degrees C. A dual channel temperature regulator can be constructed for about $ 300.

Animals