Oropharyngeal leprosy in art, history, and medicine.

Advanced lesions of the face, nasopharynx, and oropharynx have played an important role in the medical and social history of Hansen's disease. Renaissance artists included detailed portrayals of these lesions in some of their paintings, a testimony not only to their artistic skill and powers of observation but also to the common presence of these patients in European cities and towns of the period. The disease is now understood as a broad immunologic spectrum of host responses to Mycobacterium leprae, with a variety of clinical and pathologic manifestations in nerve, soft tissues, and bone. This review incorporates the findings of 2 extraordinary studies (one from Europe and the other from Japan) of pharyngeal and facial lesions. In the 1950s, studies of skeletal remains from the churchyard of a Danish leprosarium revealed a triad of maxillofacial lesions unique to leprosy and designated facies leprosa. In pre-World War II Japan, before effective treatment had been discovered, a prominent otorhinolaryngologist studying oropharyngeal and nasopharyngeal lesions prepared watercolor illustrations of the natural progression of untreated Hansen's disease. As a result of effective antimicrobial therapy, such advanced lesions are now rarely seen, but the presenting signs and symptoms of leprosy still occasionally arise in the nasal and oral mucosa. The nasopharynx and oropharynx may be important early sites of inoculation and infection by M leprae, and they require additional emphasis in worldwide efforts toward early diagnosis and treatment of Hansen's disease.

On the beta-glucuronidase binding protein (BGBP) of microorganisms. Its purification, the antiserum preparation against that and its localization in leproma and the other infectious lesions shown by immunohistologic method.

Our previous studies suggested that M. leprae (ML) grow in peripheral nerves and lepra cells because ML metabolize hyaluronic acid (HA), and use its component for their growth by the aid of host enzyme combined to the bacilli derived beta-glucuronidase binding protein (BGBP). In this study, therefore, we examined the method to purify BGBP from a mycobacterium HI-75 originally separated from a leproma and cultured by modified Ogawa's medium containing split products of HA (glucuronic acid and N-acetylglucosamine). The distribution of BGBP in leproma and the other lesions consisting of hepatitis B virus infected liver and M. avium-intracellulare infected lung tissue were also immunohistologically examined. As the result, the best method to get BGBP was preparatory electrophoresis in the final step of the purification and not the molecular sieving. The BGBP was actually proven in leproma and the other infected tissues as described, indicating the abilities of these microorganisms to utilize the metabolic machinery of the host with the similar ways to that of ML.

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.

Liberated intracellular pathogen--leprosy model.

Thirty-three mycobacterial strains, 30 by culture and 3 directly from tissues, isolated from lepromatous leprosy and leprosy infected armadillos, were compared by numerial taxonomy and by antibodies from lepromatous patients. An additional 17 strains of the M-A-I-S complex were similarly compared and all strains were compared by rabbit antibodies induced by tissue bacilli from armadillos from culture HZ-15 and by members of the M-A-I-S complex. The results are discussed in terms of the identification of M. leprae against a background of prior long-held hypotheses as to the characteristics of this bacillus.

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.

Histochemistry of B663 pigmentation: ceroid-like pigmentation in macrophages.

Histochemical studies were made of pigmented cutaneous lesions from three cases of lepromatous leprosy treated with B663 to determine the nature and histogenesis of the brown pigmentation which develops as a side effect of the drug. One case of DDS-treated leprosy and four cases of untreated leprosy were also investigated histochemically as controls. The brown pigmentation of the skin is due to deposition of a ceroid-like substance in the macrophages, which is a yellowish-brown, acid-fast lipid pigment. It is insoluble in fat solvents and accepts lipid dyes even after lipid extraction by fat solvents. The macrophages in the B663-treated leprosy contain more neutral fat and less phospholipid than the untreated lepromatous leprosy tissues. Ceroid in the macrophages probably originated from unsaturated fatty acids of the leprosy bacilli through oxidation or their binding with the drug. Crystals of the drug were not found in the macrophages in this series, even on the tissues embedded in carbowax or frozen sections.

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.