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Biomedical subjects

S S Sohi

Publications and source records attributed to S S Sohi.

16 recordsLinked to original sources

An in vitro system for testing Bacillus thuringiensis toxins: the lawn assay.

A cell assay system was developed that allows Bacillus thuringiensis delta-endotoxins activated at high pH (10.5) to be tested in vitro without causing alkaline injury to target cells. The assay is carried out on a lawn of gel-suspended cells, requires only 1 microliter of sample per dose, and is quantitative, rapid, and sensitive. The threshold dose for toxicity of B. thuringiensis subsp. kurstaki HD-73 with IPRI-CF-1 cells was 24 pg protein. The assay is also very useful for identifying antibodies which inhibit toxicity and for detecting beta-exotoxin.

Animals

Replication and serial passage of a singly enveloped baculovirus of Orgyia leucostigma in homologous cell lines.

A singly enveloped nuclear polyhedrosis virus (SNPV) of the white-marked tussock moth, Orgyia leucostigma, was successfully grown in four continuous cell lines developed from minced neonate larvae of this insect. Level of infection in two cell lines, IPRI-OL-12 and IPRI-OL-13, was 65-90%, but in the other two, IPRI-OL-4 and IPRI-OL-9, it was about 3%. Polyhedral inclusion bodies (PIBs) appeared in the nuclei of cells within 24 h postinoculation. Cytopathological changes and morphogenesis of the virus, as revealed by light and electron microscopy, were in general typical of an SNPV. However, some of the PIBs contained very few virions, and some were fractured. Rate zonal centrifugation of alkali-released occluded virions further confirmed the singly enveloped characteristics of the virus. The SNPV was serially passaged 60 times each in OL-12 and OL-13 cells. Percentage infected cells and PIB production stayed generally high throughout serial passaging in OL-13 cells, but declined sharply after the 41st passage in OL-12 cells. PIBs from the 4th passage of the virus in OL-12 cells were tested and found to be pathogenic to O. leucostigma larvae.

Animals

Bacillus thuringiensis delta-endotoxin: evidence that toxin acts at the surface of susceptible cells.

Enzymically activated delta-endotoxin of Bacillus thuringiensis covalently bound to Sephadex beads, has the same effect on insect cells in tissue culture as free toxin. The effect is prevented by antitoxin antibody and heat denaturation and is not due to a nonspecific protein effect, the beads, or toxin released from the beads. The toxin, therefore, probably acts at the cell surface.

Antitoxins

Development of an attached strain from a continuous insect cell line.

A continuous attached cell strain has been developed from the IPRI-CF-124 line of the spruce budworm, Choristoneura fumiferana. This was done by discarding suspended cells at each passage, rinsing attached cells with 0.05% trypsin and using only the strongly attached cells for subculturing. The method is very effective in that the proportion of attached cells increased from 6% in the parent cell line to 97% in the new cell strain after 20 passages. The attachment and growth properties are stable after storage of cells in liquid nitrogen. The new cell strain is designated IPRI-CF-124T and has a population doubling time comparable to that of the parent cell line.

Cell Adhesion

Chromosomal characterisation of five lepidopteran cell lines of Malacosoma disstria (Lasiocampidae) and Christoneura fumiferana (Tortricidae).

Chromosome number and morphology have been examined in four established cell lines (Md63, Md66, Md108, and Md109) of the forest tent caterpillar, Malacosoma disstria Hübner, and one (Cf124) of the spruce budworm, Choristoneura fumiferana (Clemens). Chromosome number distributions of Md63 (mode = 112) Md108 (mode = 103), Md109 (mode= 103), and Cf124 (mode = 110) overlap sufficiently to prevent identification of individual lines by number alone. However, Md66 is exceptional in possessing a modal number of 157. One large chromosome occurs in cells of all lines. The presence of this chromosome, the lack of any distinct polyploid series among chromosome numbers encountered, and the general inverse relationship between number and size of chromosomes, suggest that the high level of heteroploidy characteristic of these and other lepidopteran cell lines reflects not only a possible polyploid origin but also extensive chromosomal rearrangement and fragmentation. Tolerance for such change is attributed to the holokinetic organisation of lepidopteran chromosomes. A distinct heteropycnotic body is present in about 10% of Cf124 cell nuclei, and can be used as a marker for this line. This body may represent the sex chromatin normally encountered in somatic cells of female C. fumiferana.

Cell Line