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Biomedical subjects

O K Skinsnes

Publications and source records attributed to O K Skinsnes.

At least 19 recordsLinked to original sources

Light- and electron-microscopic study of M. leprae-infected armadillo nerves.

Lesions in peripheral nerves of armadillos experimentally infected with Mycobacterium leprae were studied by light- and electron-microscopy. Bacilli could be found clearly inside axons of unmyelinated nerve fibers. Heavily bacillated Schwann cells were seen embracing unmyelinated axons with interrupted cytoplasmic membranes. This indicated the initiation of rupture of those cells which were responsible for the liberation of bacilli into the axons. The nerve lesions were divided into three grades according to their severity: grade I showed lesions focalized in the perineurium; grade II lesions were scattered inside nerve tissue; and in grade III lesions the nerve tissues were diffusely affected. No regressive changes, such as fibrosis or scar formation, were seen in the nerve lesions. Bacillated macrophages were not as foamy as those of human lesions, indicating that these bacillated cells were younger or more easily disrupted with a higher turnover than the cells in human lesions. This would promote the spread of lesions in armadillos, and would explain the less foamy appearance of the cells. We found bacilli inside lymphatics surrounding the nerves, substantiating the opinion that lesions spread to peripheral nerves not only by a hematogenous route but also by the lymphatics.

Animals

Acid mucopolysaccharide metabolism in leprosy. 3. Hyaluronic acid mycobacterial growth enhancement, and growth suppression by saccharic acid and vitamin C as inhibitors of beta-glucuronidase.

A series of pilot studies are presented utilizing mouse and human infections with M. leprae and mouse infections with M. lepraemurium relating to the previously reported finding that hyaluronic acid seems to be a major nutrient substrate for these bacilli. The "feeding" of hyaluronic acid to the bacilli enhanced the growth of M. leprae in mouse abdominal walls and increased the Morphologic Index of M. lepraemurium infection. Saccharic acid, an inhibitor of beta-glucuronidase previously reported as present in these leprosy bacilli, caused marked regression of advanced M. lepraemurium infection, inhii. Ascorbic acid (vitamin C), also an inhibitor of beta-glucuronidase, given at a level of 1.5 gm/day for 4.5 months to one lepromatous patient without other treatment and for up to 24 months to four other lepromatous patients receiving DDS, was accompanied by lesion regression and changes in bacillary morphology similar to those seen in the inhibitor treated mice. If these observations are confirmed the possible use of beta-glucuronidase inhibitors as a useful adjunct to other leprosy therapy is raised as is also the likelihood of developing new therapies.

Animals

Cytochrome-linked respiration in host grown M. leprae isolated from an Armadillo (Dasypus novemcinctus, L.).

The bacilli were isolated from an armadillo (Dasypus novemcinctus, L.) and cytochrome systems as well as oxidation of succinate and NADH by M. leprae were studied. Cell-free extracts of M. leprae contained cytochromes of the a + a3, b, c and o type. Whole cell suspensions catalyzed the oxidation of succinate. The process was unaffected by rotenone but was markedly inhibited by thenoyltrifluoroacetone, antimycin A and cyanide. Cell-free preparations of M. leprae also oxidized NADH with oxygen as the terminal electron acceptor. Although NADH oxidation was completely inhibited by rotenone, the process was inhibited to only 50% by 5 millimols cyanide. The results indicated that complete respiratory system is present in M. leprae isolated from leprous tissues of an armadillo. The effect of inhibitors on succinate and NADH oxidations showed that the respiration in host-grown M. leprae is mediated through the cytochrome system with oxygen as the final electron acceptor.

Animals

In vitro cultivation of leprosy bacilli in hyaluronic acid-based medium. 2. Progress and developing concept of the role of hyaluronic acid suggested by culture and armadillo infection studies.

Progress is summarized relating to the verification, identification of M. leprae and understanding of the process of adaptation the pathogen passes through before in vitro growth takes place. It is recognized that hyaluronic acid apparently does not serve as a source of energy but the possibility is presented that it plays a role in the reconstruction of M. leprae cell walls made "leaky" by constant intracellular life. This apparently occurs, in culture, initially by the development of coccoid forms which after a period of weeks finally give rise to acid-fast bacilli. If these understandings are correct and the bacillary cell walls are vitated by enzyme and other action occasioned by intra-macrophage existence, then cell wall antigenicity may also be vitiated or altered by intracellular parasitism and restored by in vitro cultivation. The possible importance of this hypothesis in the understanding of immunologic responses in leprosy, and in the possibilities for therapeutic use and vaccine development are discussed.

Adaptation, Physiological

Acid-fast properties and pyridine extraction of M. leprae.

The reportedly unique pyridine extractability of acid-fastness as an identifying characteristic for M. leprae was examined in the leprosy bacilli and in eight other strains of mycobacteria. The initial findings were, in general, in accord with previous reports except that M. smegmatis and M. phlei likewise demonstrated two hour pyridine extractability of acid-fastness. Perhaps, more significantly, it was found that this characteristic in M. leprae is related to aged, probably nonviable bacilli. Some other strains of mycobacteria when tested in aged cultures showed the same phenomenon while M. leprae cultivated in vitro in a recently developed medium resisted pyridine extraction up to three weeks of growth, but thereafter as the culture aged pyridine extractability became characteristic. It is concluded that this pyridine extractability of acid-fastness is a characteristic of aging or nonviable bacilli. As such it is not definitive in the determination of whether or not in vitro cultivation of M. leprae has been achieved.

Bacteriological Techniques