Phosphoenolpyruvate carboxylase, lactate dehydrogenase, malic enzyme and aspartate aminotransferase activities were found in all strains of selected Neisseria species. Aspartase was found in all strains of the “true neisserias” except two, one N. meningitidis and one N. flavescens , but could not be demonstrated in the “false neisserias”. Some species difference in cofactor requirement was noted. Malate dehydrogenase was not found in N. meningitidis, N. gonorrhoeae , except in one strain which exhibited a trace of activity, or in N. cinerea. In the other Neisseria strains, this enzyme was present, but the activities differed markedly between the species.
Lethally irradiated mice reconstituted with allogeneic bone marrow cells usually survive the irradiation damage, but their immune-status is not completely restored. Thus, such mice show depressed humoral antibody responses to thymus dependent antigens, prolonged allogeneic skin graft survival, and their spleen cells display reduced graft-versus-host (GVH) activity when inoculated into newborn F1 hybrid mice. The present study was carried out in order to explore the reason(s) for the depressed immunological activity of allogeneic irradiation chimaeric mice. It was found that these mice had an elevated level of θ positive cells compared to mice transplanted with syngeneic bone marrow cells, but their spleen cells did not show any detectable specific or third-party GVH response. Spleen cells from allogeneic irradiation chimaeras were not able to participate in a collaborative GVH response with lymph node cells either. Possible reasons for the immunological inertness of θ positive cells in allogeneic irradiation chimaeras are discussed.
An immunofluorescence study of 119 kidney biopsies from a variety of renal affections, including transplants, is presented. The investigation was carried out with special reference to the possible presence of IgE, but simultaneously, other immunoglobulin classes (IgG, IgA, IgM) as well as complement (C 3) and fibronogen were sought for. Out of the 119 biopsies examined, 117 were negative if tested with a monospecific anti-IgE conjugate. The only definitely positive finding of IgE came from two patients with amyloidosis.
By means of the leucocyte migration technique it is demonstrated that the migration indices from 9 patients suffering from essential hypertension are separated significantly from 14 patients suffering from secondary hypertension, diabetic angiopathy, periarteritis nodosa, temporal arteritis, endarteritis obliterans and from 11 normal persons. This means that a simple secondary degenerative damage of the vascular bed is not sufficient to produce measurable cellular antibodies against the vascular wall; accordingly the positive finding in patients with essential hypertension seems to be specific and possibly an important pathogenetic factor in at least some cases of essential hypertension.
The occurrence of humoral antibodies to Strep. 2A in recurrent aphthous stomatitis (RAS) was investigated by titrating sera and by using a double-layer immunofluorescence staining method. The series comprised 24 patients with RAS and 24 persons without anamnestic or clinical signs of RAS. The distribution of endpoint titre in RAS differed significantly in relation to the distribution in the controls (p<0.005). No antibodies to a strain of Streptomyces and a strain of Neisseria used were demonstrated in RAS. The results indicate that Strep. 2A hypersensitivity is a feature of RAS.
Mice were rendered tolerant to a haptenic determinant NIP by cyclophosphamide treatment and subsequent multiple intraperitoneal injections of NIP coupled to mouse serum albumin. Control mice received no antigen. The mice were challenged with immunogenic NIP‐conjugates 20 days after stopping the tolerance inducing treatment. The response to the hapten was reduced, while the response to the carrier was normal. A second challenge showed absence of memory cell development towards the hapten. To study antibody affinity in tolerance rats were rendered tolerant to NNP‐human serum albumin and challenged later with the tolerogen. The rats developed very little anti‐NNP antibodies until 3 months after the tolerance inducing treatment. Then the affinity of antibodies was low in the tolerant group. A partial tolerance lasted one year, which is remarkably long when compared to other works on the length of B cell tolerance. Tolerant animals had only weakly specific antibody to the tolerogen in contrast to the controls. Maturation of antibody affinity was paralleled by increase in specificity in the control rats.
Radioactive protein A is actively taken up by rabbit peritoneal macrophages in the presence of immune rabbit IgG, most probably due to an immune reaction leaving the Fc sites available for macrophage adherence. The low rate of protein A uptake by guinea pig peritoneal macrophages in the presence of normal or immune guinea pig sera is apparently a result of the extensive Fc-interaction with protein A of the IgG in these sera. Delayed hypersensitivity to protein A of the donor animals does not influence the rate of uptake.
Glucokinase and glucose 6‐phosphate dehydrogenase activities are found in species of “true neisserias”, also in the non‐saccharolytic ones. The enzymes are not found in the “false neisserias” (N. catarrhalis, N. ovis and N. caviae). The stimulating effect of glucose on the synthesis of these enzymes, and the electrophoretic behaviour of glucose 6‐phosphate dehydrogenase are compared in the different species.
A method for the quantitative determination of plasma concentrations of ampicillin and cloxacillin in mixture is described. The antibiotics were separated by high voltage paper electrophoresis. The high affinity of cloxacillin for albumin seriously interfered with the electrophoretic separation of the antibiotics and the quantitation of cloxacillin. Therefore, prior to electrophoresis, the plasma proteins were digested with trypsin to release cloxacillin. Subsequent to the electrophoretic separation the antibiotics were developed micro-biologically by agar diffusion using the separated spots on the paper as diffusion centres. The inhibition zones were clearly visible after staining and could be quantitated. The method was evaluated statistically and showed good agreement with the paper disc diffusion method.
A rapid radioimmunoassay (RIA) for the measurement of hepatitis B antigen and antibody is described in which protein A carrying Staphylococcus aureus is used as a solid phase antigamma globulin reagent. The tests are easy to perform, they are completed in two hours, and are easily automatized. The competition principle is used for antigen testing while antibody is measured by direct binding of radiolabelled hepatitis B antigen. Using human reconvalescent antisera, the staphylococcal test for antigen in 25 μl serum has about the same sensitivity as the Ausria test. The staphylococcal test for antibody has at least the sensitivity of the passive hemagglutination test. Both tests are highly specific.
Seventy patients with cystic fibrosis treated as out-patients in a cystic fibrosis clinic have been followed during one year by monthly bacteriological examinations of tracheal secretions. The daily impression obtained in the laboratory is expressed by the mean point prevalence rate of Pseudomonas aeruginosa : 44 per cent, Staphylococcus aureus : 39 per cent, Haemophilus influenzae : 17 per cent, Diplococcus pneumoniae : 8 per cent, and miscellaneous other bacteria, mainly Enterobacteriaceae: 8 per cent. The fluctuations of the bacteriology are described by additional epidemiological terms: At one or more examinations during the study (period prevalence rate), 90 per cent of the patients harboured St. aureus , 64 per cent Ps. aeruginosa (mainly mucoid strains), 64 per cent H. influenzae , 37 per cent D. pneumoniae , and 30 per cent miscellaneous other bacteria, mainly Enterobacteriaceae. This pattern was found in all age groups with minor age-dependent modifications, especially as regards Enterobacteriaceae. Ps. aeruginosa was predominating as regards chronic colonization which reflects the most difficult therapeutic problems, the period prevalence rate being 39 per cent in contrast to St. aureus : 10 per cent, and H. influenzae : 1 per cent. St. aureus was predominating as regards new colonization and recolonization which reflect the problems of prevention, the incidence rate of new colonization and recolonization per risk group being 84 per cent, followed by H. influenzae : 62 per cent, Ps. aeruginosa : 43 per cent, and D. pneumoniae : 30 per cent. The results show that the bacteriological problems in cystic fibrosis are still considerable as regards therapy as well as prevention. Although many species may colonize the respiratory tracts of these patients, the main clinical problems concern Ps. aeruginosa and St. aureus ; the reason why is discussed.
A large number of bacterial strains from the upper respiratory tract were examined for production of imprints of the colonies on blood agar plates. Moraxella strains regularly grew with “corroding” colonies and most cultures produced spreading zones around the colonies after prolonged incubation. Neisseria (Moraxella, Branhamella) catarrhalis , and N. pharyngis colonies produced marked “corrosion” without spreading zones. Micrococcus mucilaginosus, Staphylococcus aureus and S. epidermidis , and some of the strains of Streptococcus pneumoniae and N. meningitidis produced slight to moderate depressions in the agar, with or without moderate “corrosion”. About 10 per cent of the strains of alpha- and beta-haemolytic streptococci failed to produce imprints on the agar. Of the remaining strains the majority produced slight to moderate depressions with or without “corrosion”. About 12 per cent of the strains produced colonies which were deeply imbedded or infiltrated in the agar. The possibility of an association of these phenomena with other physiological characters, as in some Moraxella species, is discussed.
Acta Pathologica Microbiologica Scandinavica Section B Microbiology and ImmunologyVolume 82B, Issue 6 p. 914-916 PROPERDIN ACTING AS A C3 CONVERTASE Dr. Ulf Johnson, Dr. Ulf Johnson Institute of Medical Microbiology, University of Lund, Sölvegatan 23, S-223 62 Lund, Sweden.Search for more papers by this author Dr. Ulf Johnson, Dr. Ulf Johnson Institute of Medical Microbiology, University of Lund, Sölvegatan 23, S-223 62 Lund, Sweden.Search for more papers by this author First published: September 1974 https://doi.org/10.1111/j.1699-0463.1974.tb02392.xAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat REFERENCES Cochrane, C. G. & Müller-Eberhard, H. J.: J. Exp. Med. 127: 371–386, 1968. Ganrot, P. O.: Scand. J. Clin. Lab. Invest. 29: suppl. 124: 39–47, 1972. Götze, O. & Müller-Eberhard, H. J.: J. Exp. Med. 139: 44–57, 1974. Laurell, C. B.: Anal. Biochem. 10: 358–361, 1965. Nilsson, U. & Müller-Eberhard, H. J.: J. Exp. Med. 122: 277–298, 1965. Sjöholm, A.: Scand. J. Immunol. 1974. in press. Volume82B, Issue6September 1974Pages 914-916 ReferencesRelatedInformation
The ultrastructural localization of blood group substance A and its relationship to the cell coat was studied on exfoliated buccal epithelial cells from four male subjects. The antigens were detected by a double layer immunoperoxidase staining method (IP) as well as a double layer immunoferritin (IF) staining method. The cell coat was studied using ruthenium red. There was no variation in results obtained by the IP and IF staining procedures. Positive reacting cells (IP and IF staining) showed either an even or a patchy distribution of the reaction products along the outer part of the cell membrane. Some cells had only very little or no reaction products. The cell coat appeared as an evenly distributed electron dense granular layer at a localization corresponding to that of the antigen.
A sensitive, simple and rapid method for detection of bacteriuria has been developed. By means of a Luminol chemiluminescence method adapted to an Auto Analyzer it has been possible to indicate the presence of low concentrations of bacteria in urine specimens from an unselected clinical material. Less than 1 per cent of the investigated urine samples were false negatives and 7 per cent were false positives according to the Luminol method as compared with viable count. Blood is a disturbing factor. No special treatment of the specimens of urine was needed. The analysing time was only two minutes. Formaldehyde could be added without interfering with the analysis. As the reagents required are inexpensive, and with autoanalyzers available at many hospitals, the Luminol method might be of interest as a mass screening technique for bacteriuria.
Mycobacterium ranae has been used as an experimental model to investigate the feasibility of developing a phage typing system for the subdivision of a mycobacterial species. Typing by means of unadapted phages was of limited value whereas, for one group of strains, a highly discriminative set of adapted phages was produced. The eleven phages used in the study were divisible into four groups, one consisting of three phages from pseudolysogenic strains. Two other groups have been named the Tara and Penso groups, respectively. Two hitherto undescribed phages have been isolated and designated Mx1 and Mx2.
In vitro lymphocyte activation by phytohaemagglutinin was measured as 3H-TdR incorporation by buffy coat cultures from 90 patients with sarcoidosis. Neither the median response nor the geometric mean response in sarcoidosis differed significantly from those in controls. However, when the series was divided into clinical subgroups, patients with hilar adenopathy and parenchymal pulmonary changes had significantly weaker responses. The PHA response was significantly higher in patients with erythema nodosum than in those without this symptom.
An account is given of isolation of DNA from mycobacteria. Mechanical disruption was carried out in a French pressure cell press, followed by isolation of DNA by means of Marmur's method. This DNA contained considerable amounts of polysaccharides which, however, could be removed by precipitation with cetyltrimethylammonium bromide (CTAB). The degree of purity obtained and the GC contents are given.
A colorimetric method has been tested for the determination of aminopeptidase in crude extracts of Streptococcus mitis. This was done in order to evaluate the possibility of using this enzyme as an intracellular marker in studies concerned with the quantitation of cell lysis. No purification of the enzyme was applied since the determination of cell lysis by an intracellular enzyme marker is dependent on the accurate and rapid measurement of the enzyme activity in supernatants from autolyzing cultures. Under the assay conditions used the enzymic reaction proceeded at constant rate, proportional to enzyme concentration over a wide range. The rate of the enzymatic reaction was maximal at 35–40° C. The optimum pH for activity was 7.2. The enzyme was stable in the pH‐range 5.2–9.8. The rate of the reaction was decreased at high substrate concentration.
