Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.4.21.4 (trypsin)
42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In vivo grown M. lepraemurium suspensions were inoculated into a basal medium containing cholesterol and lecithin. Slow growing strains of mycobacteria were cultured regularly in these media. The presence of free cholesterol or cholesterol in serum or cholesterol in trypsin-digested egg yolk was essential for growth. The primary cultures were difficult to obtain, but the strains were easily subcultured. A heavy inoculum was necessary to obtain primary cultures in the liquid media, no growth occurred on semisolid agar slants. Similarly slow-growing primary cultures were obtained on Ogawa egg yolk media. Growth developed in a considerably shorter time if Ogawa's medium was enriched with 0.4% yeast extract (Difco). The cultures obtained on Ogawa egg yolk media were successfully subcultered in liquid cholesterol-lecithin media. The relation of the cultured strains of mycobacteria to the pathology of murine leprosy is not yet clear. The dynamics of cholesterol metabolism in the macrophages related to murine leprosy is discussed.
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PMID:In vitro cultivation of mycobacteria in cholesterol lecithin media from lepromas of rats infected with Mycobacterium lepraemurium. 36 91

E, EA and EAC rosetting techniques and Ig fluorescence were used in a study of receptor sites in cryostat sections of lesions through the spectrum of leprosy, and for comparison in some other mycobacterial and granulomatous lesions. Anti-C3, and trypsin were used as blocking agents. Lymphocytes in borderline lepromatous leprosy produced EA adherence and IgG fluorescence indicating B type cells. Lymphocytes in tuberculoid leprosy produced neither E or EA adherence and no fluorescence; these cells were presumed to be T cells. EAC and EA adherence was more marked in areas of macrophage infiltration, where there were few lymphocytes, than over the lympocytes themselves. Two distinct patterns emerged: (i) EA binding together with IgG fluorescence was seen in active lepromatous leprosy and could be localised to the surface of individual macrophages, and (ii) EAC binding together with IgM fluorescence was seen in the granuloma of tuberculoid leprosy and sarcoidosis, but could not be definitely related to cell surface; rather it was diffusely spread over the whole granuloma; EAC adherence was diminished by anti-C3 serum. Trypsin removed EA binding completely, but only diminished EAC adherence. It is suggested that the EA pattern indicates immunoglobulin receptors on macrophage and lymphocyte surfaces: and that the EAC binding (which is stronger than EA) involves C3 and IgM receptors at extracellular sites as well as C3 receptor sites on epithelioid cell surfaces. EA and EAC binding were enhanced in borderline tuberculoid leprosy in reaction and erythema nodosum leprosum, suggesting that immunoglobulin and complement receptor sites increase in number with enhanced hypersensitivity.
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PMID:Surface markers on lymphocytes and cells of the mononuclear phagocyte series in skin sections in leprosy. 72 93

Acid-fast bacilli multiplied in liquid culture media containing hyaluronic acid when inoculated with mycobacteria from a lepromatous leprosy nodule. The culture was readily subcultured at ten day intervals in the homologue media, but failed to grow in the Dubos, Middlebrook and Lowenstein media. These findings confirm the results of Skinsnes et al (1975). Identification of this culture is not yet available, however it gives positive immunofluorescence with authentic anti-M. leprae serum. The obtained culture also grows as a chromogenic culture at 34 degrees C on a simple medium prepared from trypsin digested human umbilical cord, yeast extract powder and glycerol. This medium can be sterilized in an autoclave, but filter sterilized sheep, bovine or horse serum must be added aseptically as an essential ingredient. The medium does not differ considerably from the hyaluronic acid medium proposed by Skinsnes et al, but it is easier to prepare, it is inexpensive and permits a logarithmic growth within seven days of the so far unidentified culture isolated from leprotic nodules.
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PMID:A simplified hyaluronic acid based culture medium for mycobacteria isolated from human lepromata. 79 28

The oxidation of 3,4-dihydroxyphenylalanine (DOPA) was studied by spectrophotometric methods at pH 6.8. In the presence of L- or D-DOPA, a color development occurred in the presence of the following substances as measured by increase in absorption both at 540 nm and 480 nm: hyaluronic acid, trypsinized human skin and umbilical cord extract, trypsin treated rat tissue from subcutaneous rat leproma, trypsin treated M. lepraemurium isolated from rat lepromata, and trypsinized M. leprae isolated from non-treated lepromatous leprosy cases. Normal human skin and connective tissue extract and nontrypsinized connective tissue of rat leprosy granuloma did not oxidize DOPA. While the trypsin-treated partially purified M. leprae suspension oxidized DOPA at both wave-lengths, the hyaluronidase-treated same suspension of M. leprae failed to oxidize these phenolic compounds. Mushroom tyrosinase oxidized D-DOPA, L-DOPA, epinephrine and norepinephrine at 480 nm. Hyaluronic acid also oxidized epinephrine and norepinephrine at both wave-lengths. Since it is known that M. leprae in the human host is closely associated with the presence of the acid mucopolysaccharides of the skin, and since acid mucopolysaccharides and skin constituents strongly oxidized DOPA, and since the hyaluronidase treated M. leprae failed to oxidize DOPA, it became evident that hyaluronic acid and not M. leprae is responsible for DOPA oxidation, and phenolase activity is not associated with the metabolism of M. leprae. Evidence is presented that DOPA is not a unique characteristic of the human leprosy bacillus. For instance, trypsin-treated murine leprosy bacilli from the rat strongly oxidized DOPA. The reaction of DOPA oxidation, therefore, must be rejected as a test for the identification of M. leprae. The obtained results confirmed the pertinent findings of Skinsnes and his co-workers.
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PMID:Oxidation of 3,4-dihydroxyphenylalanine by connective tissue constituents. Identification of Mycobacterium leprae not related to phenolase activity. 82 25

We prepared antigens by precipitating with 80% ammonium sulfate supernatants of human and armadillo antigen at a concentration of 160 X 10(6) bacteria per ml. The precipitate was resuspended, dialyzed and filtered. The antigen obtained was inactivated with trypsin during 30 minutes. The tests made with these antigens were negative for the 48-hour test in lepromatous patients and highly positive in normal persons who were contacts of leprosy patients.
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PMID:Comparative study of the 48-hour response to soluble antigens obtained from human and armadillo leprosy material in lepromatous leprosy patients and normal persons, contacts of leprosy patients. 94 41

Two antigens were tested and compared in relation to the 48-h Fernandez reaction. They were obtained from standard human and from standard armadillo lepromin. All the tests were negative in patients with lepromatous leprosy and highly positive in those with tuberculoid leprosy and in lepromin-positive contacts. There was total agreement in all tests done with the two types of antigen. The antigenic component has the following basic properties: it precipitates with 80% saturated ammonium sulfate; it is not destroyed by autoclaving or by treatment with 0.4% phenol; it is non-dialysable; and it is destroyed by treatment with trypsin.
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PMID:Specificity of the 48-hour reaction to Mitsuda antigen. Use of a soluble antigen from human and armadillo lepromin. 108 6

A microagglutination test using trypsin-treated and Coomassie blue-stained Trypanosoma cruzi epimastigote antigen was adapted for the diagnosis of Chagas' disease. When incorporated in the test, 2-mercaptoethanol treatment of chagasic sera had no influence on antibody titer. In contrast, titers in sera from patients with visceral leishmaniasis, African trypanosomiasis, and autoimmune disorders, subjected to similar treatment, showed remarkable decline. Accordingly, a lower cut-off point for Chagas' disease serological negativity could be taken resulting in a higher sensitivity (95.6%); the specificity was 94.7%. Similar specificities were obtained with Leishmania donovani chagasi and L. d. donovani antigens applied to homologous visceral leishmaniasis and heterologous Chagas' sera. Of 316 nonchagasic sera, only 3 with leptospirosis and 1 with leprosy showed seropositive titers prior to and after 2-mercaptoethanol treatment.
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PMID:Trypsin-treated and coomassie blue-stained epimastigote antigen in a microagglutination test for Chagas' disease. 244 Mar 29

Because the correct diagnosis of indeterminate leprosy (IL) requires the finding of acid-fast bacilli in skin lesions from clinically and histopathologically suggestive cases, it is necessary to develop a reliable method for this purpose. This paper presents a simple procedure, available to every general laboratory, which consists in obtaining 2 suspensions: SI, by mincing and grinding the tissue in phosphate-buffered saline; and SII, after treating SI with NaOH solution and digesting with trypsin. In 22 IL skin biopsies, bacilli were directly observed in only 3 with the Ziehl-Neelsen (ZN) stain; and with the peroxidase-antiperoxidase method it was impossible to differentiate between nonspecific precipitate and true positive reactions. In contrast, 18 positive results from the same 22 samples were obtained when both SI and SII were evaluated with ZN stain. The logarithmic bacterial index was also increased in at least 7 cases.
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PMID:Demonstration of acid-fast bacilli in skin biopsies from indeterminate leprosy cases. 306 61

Macrophage cultures pulsed with viable Mycobacterium leprae were assessed for erythrocyte rosetting in three groups of individuals, i.e., normal subjects, and tuberculoid and lepromatous patients. Of these, only the lepromatous group showed a reduction in rosetting ability after infection with M. leprae. The specificity of such a reduction pattern was confirmed by using various mycobacteria to infect the macrophages. A threshold effect was noted in all three groups. Although a reduction was obtained in the amount of rosetting of macrophages from lepromatous patients with 10(4) acid-fast bacilli per culture, tuberculoid and normal macrophages resisted such an effect with as large a dose as 20 X 10(6) to 30 X 10(6) and 30 X 10(6) bacilli per culture, respectively. The M. leprae-caused alterations in macrophages from lepromatous patients were reversible by treatment with trypsin and colchicine. Cytochalasin B and Tween 80 were unable to alter the pattern. Treatment of cells with neuraminidase was inconclusive since it enhanced rosetting values of both control and infected cultures. These manipulations were significant in elucidating the target point of the host (macrophage) and parasite (M. leprae) interaction and in delineation of the external and internal effects upon the macrophages. Both M. leprae and macrophages were participants in Fc reduction, as treatment of the former with rifampicin and of the latter with cyclocheximide significantly augmented the rosetting ability. In conclusion, it appears that M. leprae, upon entering a lepromatous macrophage, initiates the production of a protein which acts via the microtubules to alter membrane topography. It is possible that the altered membrane prevents effective macrophage-lymphocyte interaction. This could be one of the mechanisms by which cell-mediated immunity is suppressed in lepromatous leprosy.
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PMID:Alterations in the membrane of macrophages from leprosy patients. 634 87

Peripheral blood derived macrophages from lepromatous leprosy patients were unable to interact with lymphocytes in the presence of M. leprae. This lack of interaction is probably not associated with membrane HLA-Dr antigens since trypsin and colchicine restored M. leprae induced depression in the latter but were unable to bring about a positive interaction. Two possible defects exist therefore in the lepromatous macrophage. These are an innate inability to process and present M. leprae antigens to lymphocytes and an induced inability to express some membrane receptors, an event detrimental to the normal functioning of a macrophage.
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PMID:Antigen specific macrophage-lymphocyte interaction in lepromatous leprosy. 637 10


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