Zentralblatt fur Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene. Erste Abteilung Originale. Reihe A: Medizinische Mikrobiologie und Parasitologie最新文献

Eleven pyrimido-pyrimidine derivatives, seven with significant antiviral activity against Mengovirus, five against Coxsackie B1 virus and four antiviral negative compounds were tested for their photosensitizing ability. All seven compounds with antiviral activity in vitro showed an enhanced antiviral action against Mengovirus under irradiation with visible light, a fact that may be caused by photodynamic processes. It was tried to correlate the oxidation potentials of sensitizers with their photodynamic activity. By means of mass-spectrometric investigations, molecular fragmentation was examined following thin layer chromatography (TCL) before and after irradiation. Furthermore, binding affinity to biopolymers (BSA and RNA) was investigated to reveal conformity in differences of antiviral activity. The main results are the following: 1. Generally, strong antiviral activity can be correlated with strong binding affinity. 2. No significant correlation could be detected between oxidation potentials of antiviral compounds and their enhanced antiviral activity under irradiation conditions, although in some cases sensitizer with higher oxidation potentials are more effective than those with lower ones. 3. The lower the photostability of the compounds the higher was the light-induced antiviral activity. 4. No alteration of the molecular ion peak and fragmentation pattern before and after irradiation was indicated by means of mass-spectrometry and TLC using fairly comparable conditions.

A simplified tube O-agglutination test was developed and evaluated for the determination of somatic serogroup (O) antigens of Serratia marcescens. Use was made of Tryptic Soy broth (TSB)-grown O-cells that had been boiled for 1 hour; 0.145 M NaCl proved a satisfactory diluent. Various technical parameters of this test were examined as well. Rabbit anti-O immune sera, that had been elicited with 5 x concentrated, TSB-grown, 1 hour-boiled O-cells of all 15 currently employed O-antigen reference strains of S. marcescens yielded satisfactory O-agglutinin titers. The tube O-agglutination test compared favorably with the indirect hemagglutination technic, although the latter technic yielded significantly higher O-agglutinin titers with merely 7 of the 15 O-antigens of S. marcescens. The tube O-agglutination test permitted detection of higher O-agglutinin titers than a microtiter O-agglutination test utilizing O-cells that had been stained with safranin O. Conversely, titers obtained with TTC-stained O-cells in a microtiter agglutination procedure approximated those yielded by the tube O-agglutination test, but O-cells of the various S. marcescens strains were stained nonuniformly by triphenyltetrazolium chloride (TTC). As before, there existed marked serologic cross-reactivity between O-antigens O6 and O14. A new O6 candidate strain, S. marcescens isolate S 1i, serotype O6:H20, was proposed. Contrary to O-agglutinins of human control serum, the O-agglutinins of rabbit anti-O immune sera proved refractory to treatment with 0.1 M of 2-mercaptoethanol (2-ME) and 0.01 M dithiothreitol (DTT) respectively. Dual absorptions of rabbit anti-O immune sera with killed cells of Staphylococcus aureus Cowan I (protein A), failed to significantly reduce O-agglutinin titers, although human IgG and IgM was bound by protein A. It was tentatively concluded that the 2-ME- and DTT-refractory rabbit anti-S. marcescens O-agglutinins resided in the IgM immunoglobulin class.

The Bacillus cereus protein has been obtained from culture fluid in homogenic form as indicated by SDS-disc electrophoresis and immunodiffusion not described before. The protein has a molecular weight of 100000 daltons. Purification was accomplished by the following steps: (1) removal of ballast nitrous components with DE-32 cellulose at pH 7.2; (2) removal of the proteins from the culture filtrate (deluted four times by water) with DE-32 cellulose at pH 8.6; (3) elution by 0.005 M tris-HCl buffer at pH 7.0 containing 0.5 M NaCl; (4) column rechromatography on DE-32 cellulose at pH 8.6. The isolated protein was identified as a vascular permeability factor acording to the bluing zone in rabbit skin tests or to the bluing lung tissue in mice.

Swine fever is conceived as a disorder of the enzyme systems, that are controled by serine proteases. The virus is replicated in the cells of the lymphomycoid complex, whereby the production of a chymotrypsin is induced. In swine fever the lymphatic glands and the lymph flow are increased. Fifteen normal pigs had chymotrypsin contents in the lymph of the body lymphnodes of 0,4 U/l, nine pigs suffering hog cholera 1,5 U/l. In the intestinal lymphnodes the chymotrypsin concentration was normally 2,9 U/l and in swine fever 3,5 U/l. Chymotrypsin which is present may induce the production of further chymotrypsin. Fourteen pigs suffering from swine fever showed increased peritoneal fluids (50 to 120 ml), whereby chymotrypsin was found in 5 cases. The lymphflow was assumed to be five times higher, when compared to control animals. This entails a seven-fold increase of chymotrypsin which enters the blood stream. In some cases the virus titers are higher in the lymph specimen and peritoneal fluids than in the serum. Increase of chymotrypsin concentration reduces the resistance of the virus in the lymph. Obviously the virus is spread in the body migrating with the lymph flow. However, the increasing chymotrypsin concentration seems to inactivate the virus and lymph retains its defense character. Detection of the fluorescent antigen is correlated with the evidence of the proteolytic precipitating antigen. After infection with the virus of swine vesicular disease increase of chymotrypsin is also evidenced in the lymph but to a lower degree. Therefore in swine fever the lymphnodes cause chymotrypsin formation to an extent which may explain the pathophysiological disorders in those physiological systems, that are controled by serine proteases.

By use of 6-molar guanidinium chloride a potent Coxiella burnetii antigen could be produced for diagnostic purposes from infected yolk sacs, with little technical expense. This treatment did not only a cause remarkably purifying effect (Tab. 1) but also the extraction of soluble cytoplasmic substance. From guanidinium chloride-treated suspensions a highly purified and uniform suspension of cell walls could be separated by Saccharose Density Gradient centrifugation (Fig. 2). Guanidine extracted organisms retained their full antigenic potential with respect to Phase I and Phase II and lacked anticomplementary activity. Such preparations can be used for serological tests like complement fixation reaction or Enzyme Linked Immunosorbent Assay and are particularly suitable for biochemical studies of Phase antigens of Coxiella burnetii.

The authors investigated the procedure which demonstrates, by means of the latex agglutination test (LA) the residual IgM antibody activities in sera preabsorbed with protein A-bearing Staphylococcus aureus and evaluated the usefulness of this technique with the following results. (1) LA (Toxotest-LA, Eikenkagaku Co.) gave clear-cut and reproducible agglutination patterns. Its specificity was comparable to that of HA and its sensitivity was somewhat lower than that of HA. (2) The absorption of 0.2 ml of 1:10 dilution of many test sera with the sediment of 0.5 ml of a 10% bacterial suspension for ten minutes or more provides the suitable condition for the absorption procedure. (3) HA-positive 183 sera revealed no residual antibody activities after absorption. These are supposed to derive from persons with a long-standing infection. Sera from 4 laboratory infection cases, from 2 toxoplasmic lymphadenopathy cases and from a pregnant woman, showed more or less residual activities. In 5 out of 7 sera tested, the residual activities were 2-ME sensitive, indicating the presence of specific IgM antibodies. (4) The specific and the total IgM were reduced slightly by the absorption procedure.

A long term study with bivalent live influenza vaccine was carried out in 18 subjects with no previous history of egg protein hypersensitivity. Experimental conditions included a nine-fold vaccination schedule with collection of serum and nasal fluid. The parameters studied were determination of serium and local antibody formation as well as the demonstration of specific IgE antibodies in serum and nasal fluid. Our major interest was directed towards the question of potential sensitization after repeated doses of non-purified oral vaccine. The close medical follow-up of the subjects revealed no clinical signs of atopic reaction. There were no complaints regarding adverse reactions usually following local application of live influenza vaccines. Determination of total serum IgE rederately elevated levels before, during und after the trial; one vaccinated subject showed high concentrations prior to vaccination with no significant change during the experiment. That individual was ultimately classified atopic with a pronounced hypersensitivity to egg protein. Nevertheless this person tolerated nine doses of vaccine without side reactions and showed no significant increase in total or specific IgE antibodies. Concentrations of IgE in nasal secretion of non-atopic subjects are less than 2U/ml, whereas they are frequently higher in allergic patients and in the presence of nasal IgE levels greater than 4 U/ml one would expect a specific reaction to challenge allergens. In our vaccinees nasal IgE values were consistently within normal range, at no time exceeding 2.6 U/ml, even the atopic subject did not exhibit higher levels in nasal fluid. A correlation between systemic and local IgE antibodies revealed no pathognostic relations; in addition to this, specific IgE-serum-antibodies as measured in the RAST against ovalbumin allergen did not show any correlation to vaccination. These data present good evidence for the innocuity of the vaccine with regard to its repeated application in man.

The influence of Nifurtimox on Trypanosoma brucei, T. congolense, T. equinum, T. equiperdum, T. evansi, T. gambiense, T. lewisi, T. rhodesiense, T. vivax, Leishmania donovani and L. tropica has been studied in animals and in vitro. The drug was active against all of them but there are considerable differences in sensitivity of the various species as well as of different strains of T. rhodesiense. For the treatment of T. rhodesiense a single high dosage was more efficient than the same dose divided into many smaller applications. The latter dose schedule is more suitable for the treatment of T. cruzi and T. lewisi. L. donovani and L. tropica responded to Nifurtimox but only in vitro.

A survey was made on the incidence of coccidial oocysts in 11,365 fecal samples from Japanese broiler houses during five years from 1973 to 1977. The Eimerian species were identified by a combination of two methods, examination of oocyst morphology and chicken passage test. Oocysts were detected from 7,526 (66.2%) of the samples tested during five years. Annual variation was statistically significant at the 5% level. Coccidial incidence during July to September (corresponding to summer season) was found to be higher than that of other seasons. With regard to regional variation, incidence found in the north-eastern half of Japan was lower than that in the south-western half. Oocyst detection rate increased up to 40 days of age in chickens and kept plateau thereafter. Eimeria acervulina predominated throughout the years, seasons, regions and age of chickens. Other species of Eimeria were subjected to wide fluctuation while the survey was carried out.