The nature of molybdenum-cofactor.

In vitro assembly of Neurospora crassa NADPH-nitrate reductase (EC1.6.6.2) could be effected by combing the nitrate induced Neurospora crassa mutant nit-1 with the extract of any known molybdenum-containing enzyme. The process involves the participation of a molybdenum-cofactor contributed by the molybdenum-enzyme fraction. This paper emphasizes two points: Firstly, the indispensable role played by EDTA in the viability of Mo-cofactor and secondly, the nature of Mo-cofactor predicated by our previous work is supported by concrete experimental results. Recent experiments with Chelax-100 column provide evidence that the in vitro formation of Neurospora NADPH-nitrate reductase involves EDTA and the latter may take part in the formation of a molybdenum, labile sulfide and EDTA complex. In addition to 10(-2) M sodium molybdate, both EDTA and reducing agent are required to activate the cofactor in the Chelax-100 column eluate. The cofactor is of low molecular weight and devoid of protein as was predicated. To substantiate those predications, concrete experimental results are provided.     

The physical nature of life.

Life evolved from the primeval world of physics. Sensory systems inform animals of the natural environment, enabling them to conduct responsively. The discovery of weak, DC bioelectric fields in the vicinity of aquatic organisms and the role they play in guiding sharks and rays to their prey have led to the recognition of fundamental, hitherto less well known, physical aspects of sensory biology. The inferred cybernetic algorithm of electric-field orientation in sharks and rays is highly effective and extremely robust. In orienting to the weak DC electric fields of ocean currents and to the earth's magnetic field, sharks and rays unwittingly practise the motional-electric principles that Einstein had in mind when he introduced the special theory of relativity. At the sense-organ, receptor-membrane, and ion-channel levels, the elasmobranch ampullae of Lorenzini operate on the basis of graded positive feedback driven by negative conductance, supposedly employing voltage-sensitive ion channels as the active, excitable elements. The electric sense of sharks and rays presents an exquisite implementation of the very biophysical principles that also govern the graded, much richer than all-or-none, integrative brain processes of animal and man.     

Evidence for the nature of the link between the arabinogalactan and peptidoglycan of mycobacterial cell walls

The long-posed question of the nature of the link between the mycolylarabinogalactan and the underlying peptidoglycan of the cell walls of Mycobacterium sp. has been addressed. The insoluble cell wall matrix of Mycobacterium leprae, Mycobacterium tuberculosis, and Mycobacterium bovis was partially hydrolyzed with acid either before or after per-O-methylation and the resulting oligosaccharides further derivatized and analyzed by gas chromatography/mass spectrometry. The structures of fragments arising from the reducing end of arabinogalactan demonstrated the existence of the terminal sequence----5)-D-Galf-(1----4)-L-Rhap-(1---3)-D-GlcNAc. Other analyses confirmed the presence of muramyl-6-P within the peptidoglycan of these mycobacteria. Based on the acid lability of the 3-linked GlcNAc unit, the presence of about equimolar amounts of Rhap-(1----3)-D-GlcNAc and muramyl-6-P in an isolated cell wall fragment, and 31P NMR analysis, it was concluded that the GlcNAc residue of the terminal triglycosyl unit of arabinogalactan is joined by 1-O-phosphoryl linkage to the 6-position of some muramyl residues within the peptidoglycan. Thus, it is reasoned that the massive mycolylarabinogalactan of mycobacteria, responsible for aspects of disease pathogenesis and much of the antibody response in infections, is attached to the peptidoglycan framework by the actinomycete-specific diglycosylphosphoryl bridge, L-Rhap-(1----3)-D-GlcNAc-(1----P, perhaps thereby providing a unique target for site-directed chemotherapy of mycobacterial infections.     

The evolution of immunosuppressive cell populations in experimental mycobacterial infection.

Immunosuppressor activity of considerable potency and complexity was generated during the course of chronic, progressive infection of C3H/Anf mice by Mycobacterium lepraemurium. From the 5th through 10th week after inoculation, spleen cells from infected mice mildly but reproducibly suppressed the direct plaque-forming cell response of normal spleen cell cultures to sheep erythrocytes. Suppression at this stage of infection was mediated by cells with macrophage-like characteristics. A marked increase in splenic suppressor activity at 10 to 11 weeks was associated with the appearance of a second suppressor cell subpopulation composed of T lymphocytes. The appearance of these cells was closely related in time to the onset of rapid splenic enlargement and a loss of cutaneous delayed type hypersensitivity to antigens of M. lepraemurium in mice at 10 to 11 weeks of infection. Suppressor cells were not present in peripheral lymph nodes until terminal infection at 22 to 25 weeks. Suppressor spleen cells depressed the T-dependent antibody response most severely, but there was also a direct effect upon B cells as shown by moderate suppression of responses to TNP-LPS and DNP-Ficoll. Spleen cells from 14-week-infected mice generated a soluble suppressor factor(s) that induces depression of moderate severity, however, the immunosuppression by intact cells was far greater.     

Clinical aspects of atypical mycobacterial infection.

Summary: Atypical mycobacterial infections may give rise to various clinical difficulties. Case reports of six patients--three adult patients with pulmonary lesions, two children with cervical lymphadentis and one patient in whom the atypical mycobacterium appeared to be present as a commensal--illustrate these difficulties. Determination of the significance of the organism and differentiation of condition from tuberculosis and others requires consideration of the clinical picture, the results of skin testing, histologic features and cultural characteristics. Three patients, including the two children, were treated with a combination of surgery and chemotherapy, with satisfactory results. An elderly patient with chronic bronchitis and a pulmonary infection due to M. kansasii was treated successfully with antiuberculosis agents alone. Chemotherapy is being tried on a fifth patient with cavitary disease due to M. intracellulare, but is seems likely that an operation will also be required.     

Superficial mycobacterial lymphadenitis in Saskatchewan.

A total of 43 bacteriologically verified cases of superficial mycobacterial lymphadenitis were reported in Saskatchewan between 1981 and 1986; 35 (81%) were due to Mycobacterium tuberculosis. Among the eight cases (19%) due to nontuberculous mycobacteria the agent most frequently isolated was M. avium-intracellulare. Five additional cases were smear-positive and culture-negative. Direct smears of node tissue or aspirate were positive for acid-fast bacilli in 7 (88%) of the 8 cases of nontuberculous mycobacterial lymphadenitis but in only 16 (46%) of the 35 cases due to M. tuberculosis. Superficial tuberculous lymphadenitis was most frequent in female North American Indian or Asian-born adults and most commonly involved the cervical nodes. Nontuberculous mycobacterial lymphadenitis was most frequent in female white children, and most commonly involved the submandibular nodes. The cases of both tuberculous and nontuberculous mycobacterial lymphadenitis were spread throughout the province. There was an urban concentration of cases of tuberculous lymphadenitis in those of Asian origin. It is important to distinguish between superficial mycobacterial lymphadenitis due to M. tuberculosis and that due to nontuberculous mycobacteria for treatment and management purposes.