Botulinum antitoxins and antibacterial IgM and IgG antibodies in sera of persons immunized with botulinum polytoxoid combined with cholera vaccine. I. Response to botulinum toxoid.

Two groups of subjects were immunized with combined vaccine containing aluminum hydroxide-adsorbed botulinum toxoids, types A, B, and E, and 4 mld of formaldehyde-inactivated V. cholerae Inaba and Ogawa organisms. The first group included laboratory workers who were previously immunized against cholera and had professional contact with botulinum toxins and viable V. cholerae organisms. The second group included young men who were never vaccinated against botulism or cholera. The three-dose immunization schedule with combined vaccine resulted in clear-cut antitoxin response; after the third dose, the A, B, and E antitoxin level ranged from 0-2 to 10 IU/ml. Immunity against botulinum toxins lasted at least one year. Distribution of the antitoxins among IgM and IgG globulin classes resembled that in the case of response to other toxoids; 21 days after the third immunization antitoxin activity was found in IgG globulins.

Botulinum antitoxins and antibacterial IgM and IgG antibodies in sera of persons immunized with botulinum polytoxoid combined with cholera vaccine. II. Response to cholera vaccine.

The response of humans to cholera vaccine was very heterogeneous. The proportion of IgG vibriocidal antibodies was high in persons having previous natural or artificial contact with V. cholerae antigens. Predominance of IgM antibody response was seen in persons vaccinated for the first time. This type of response was sometimes evoked by unspecific stimuli such as botulinum polytoxoid without cholera vaccine. Antibodies passively protecting mice were found both in IgM and IgG globulins but the activity of these antibodies was higher in IgG than in IgM globulins.

Dissolution and relative bioavailability of two carbamazepine preparations for children with epilepsy.

The in-vitro dissolution profiles of two carbamazepine formulations (Tegretol and a generic carbamazepine) have been assessed and the bioavailability of carbamazepine compared in 12 epileptic children at steady-state. Dissolution from the generic formulation (100 and 200 mg tablets) tended to be greater than for the proprietary tablets. However, the bioavailability and pharmacokinetics of carbamazepine when assessed at steady-state were similar for the two formulations. It appears, therefore, that the breakthrough seizures and higher incidence of neurological side-effects observed when children were given generic carbamazepine in place of the proprietary formulation cannot be accounted for by differences in bioavailability or pharmacokinetics.

Effect of adsorbents on IgM and IgG measles antibodies.

Sera from rabbits immunized with L-16 measles virus absorded with monkey blood cells; kaolin and blood cells; and MnCl2 and heparin were examined in haemagglutination inhibition (HI) and neutralization tests. Kaolin and MnCl2 adsorbed primarily HI IgM antibodies from the early immunization period. The adsorbents used had no influence on HI and neutralization IgG antibodies. Human convalescent serum gave similar results, i.e. only IgG antibodies were found and they were not affected by kaolin and MnCl2 with heparin.

Measles antibodies in nasal secretions and sera of children with measles.

Measles antibodies were determined, in the course of measles, in sera and nasal secretions of 54 and 27 children, respectively. The examinations were performed on the 3rd or 4th day (1st period) and between the 10th and 14th day (2nd period) after onset of rash in both sera and nasal secretions and after 25 to 60 days (3rd period) in sera only. Geometric mean titres of antibodies in sera determined by haemmagglutination inhibition (HI) and neutralization tests in the 1st period were 126.3 and 115.3 respectively. For the 2nd period the respective figures were 318.4 and 396.1 and for the 3rd period--388.0 and 445.6. Fractionation on Sephadex G-200 of sera from the 1st period revealed HI and neutralizaing antibodies associated mainly with the IgM serum globulin class. Measles HI and neutralizing antibodies were also found in nasal secretions of all 27 children, but their titres were much lower than in serum. The antibodies determined by indirect immunofluorescence in nasal secretions were associated with IgA in 26 and with IgG immunoglobulin in 15 of the 27 subjects. No IgM antibodies were found.

[Drug resistance of Enterococcus species isolated from the urogenital system]. / Lekoopornosc szczepów Enterococcus sp. izolowanych z ukladu moczowo-plciowego.

Despite low virulence of enterococci, they have become important nosocomial pathogens. This has been correlated with the increased use of broad-spectrum antibiotics, particularly cephalosporins. Many strains of enterococci exhibit multiple drug resistance; the most important being high-level resistance (HLR) to penicillin (MIC > 100 mg/l) and gentamicin (MIC > 500 mg/l and 2000 mg/l) and/or streptomycin (MIC > 2000 mg/l). The investigation was performed on 92 strains, isolated from genito-urinary tract and recognised as Enterococcus sp. All strains were obtained from several microbiological laboratories of Gdansk, Gdynia and Tczew. On biochemical reaction profiles species of enterococci were identified as: E. faecalis (72.8%), E. faecalis varians (9.8%), E. durans (7.6%) and E. faecium (9.8%). The minimal inhibitory concentration (MICs) of penicillin, ampicillin, azlocillin, imipenem, gentamicin, amicacin, ciprofioxacin and vancomycin were determined by the agar dilution method. None of these 92 enterococcal strains was vancomycin resistant. 22.2% of E. faecium and 7.5% of E. faecalis showed high-level resistance to penicillin. None of these strains were produced beta-lactamase. High-level resistance to streptomycin and gentamicin was detected. Both--high-level resistance to streptomycin and gentamicin--were found in 6% E. faecalis; 11.1% E. faecalis varians and 22.2% E. faecium.

[Migration of Al(OH)3 in the bodies of experimental animals]. / Migracja Al(OH)3 w organizmie zwierzat doswiadczalnych.

The aim of this study was to observe aluminium hydroxide distribution in mice given this substance subcutaneously and intraperitoneally in various concentrations (0.03 mg-0.8 mg of Al3+ per dose). Each experimental group consisted of 5-6 mice of which brain, liver and kidney were isolated after 7, 14, 28, and 35 days after injection. Control group was composed of unvaccinated mice. Sera of animals vaccinated subcutaneously with the following doses: 0.2 mg, 0.4 mg and 0.8 mg were also tested. A distinct accumulation of Al3+ in liver after intraperitoneal injection with a tendency of rising after 28 days of observation was seen. The same was observed in kidneys after subcutaneous injection, especially doses of 0.4 mg Al3+ and 0.8 mg Al3+. The aluminium content in sera was high in an early period of observation only, and subsequently its elimination was fast.

[A rapid method of measuring the level of aluminum in biological preparations using aluminon]. / Szybka metoda oznaczania glinu w biopreparatach za pomoca aluminonu.

The aim of this study was to adapt aluminon for determination of Al3+ content in biopreparations adsorbed on Al(OH)3. Aluminum + aluminon complex was identified by spectrophotometry at A535. It was found that the method applied allows to obtain reproducible results. Its sensitivity for Al3+ contains between 85 to 680 micrograms. This method is less laborious in comparison with so far used versenian method.

[Examination of protein MPB 70 properties in BCG substrains used for the production of Polish antituberculosis vaccines]. / Badanie obecnosci bialka MPB 70 w podszczepie BCG uzywanym do produkcji Polskiej szczepionki przeciwgruzliczej.

The aim of the study was examination of MPB70 protein production by different BCG substrains and testing this protein as reagent in allergenic skin test on BCG vaccinated guinea pigs. Three BCG substrains: Danish 1331 (D), Japanese 172 (J) and Polish BCG Moreau (P) were used for the study. The protein MPB70 received from dr Nagai from Osaka University was used as reference preparation. It has been shown that MPB70 protein of molecular weight about 21,500 Da was present only in BCG Moreau (P) and Japanese (J) substrains. In immunoblotting test monoclonal antibodies reacted additionally with 43,000 Da protein in all tested substrains. We suspect that the detected additional molecule was a dimer of MPB70 protein. The development of skin reactions to MPB70 protein was seen in guinea pigs vaccinated with J and P substrains. The peak of alergy to MPB70 protein was observed 9 weeks after vaccination. In sera of vaccinated animals antibodies against MPB70 protein were detected by ELISA method. The possibility of using MPB70 protein for diagnostic purpose is discussed.

[Evaluation of prekallikrein activator (PKA) activity in plasma-derived preparations]. / Ocena aktywnosci aktywatora prekalikreiny (PKA) w preparatach osoczopochodnych.

The study was carried out on 111 series of human immunoglobulin preparations (81 series for intravenous use, 30 series for intramuscular use) produced abroad and in Poland. Kallikrein (KK) and prekallikrein activator (PKA) activities were identified using chromogenic substrate. S-2302 and proteinase inhibitors were tested by double immunodiffusion method. Several biochemical, biological and serological methods were also applied which are valid in the control of intravenous and intramuscular immunoglobulin preparations. It was found that trace amounts of PKA and KK are present in human immunoglobulin preparations for intravenous use such as Bioglobulina and Gamma-Venina (PKA 0.0092-0.0505 mu kat/L, and KK 0,-0.0093 mu kat/L). On the other hand normal human immunoglobulin preparations for intramuscular use had somewhat higher level of KK and PKA and anti-HBs Immunoglobulin preparations (5.50 mu kat/L and 153.91 mu kat/L) and Tetabulina preparations had very high levels of these factors as compared to remaining preparations (4.50 mu kat/L and 316.5 mu kat/L).

[Level of bacterial endotoxins in Haemophilus influenzae type b vaccines conjugated with toxoids (td, Di)]. / Poziom endotoksyny bakteryjnej w szczepionkach Haemophilus influenzae typ b skoniugowanych z toksoidami (Te, Di).

Quantitative evaluation of bacterial endotoxin was performed in the following vaccines: Act-Hib, Hiberix, Hib-Titer. The aim of this study was to assess the accuracy and precision of chromogenic LAL test with S-2423 substrate for this particular biopreparations and after that to determine the amounts of endotoxin as a factor of vaccine safety. Because of the lack of information concerning the presence of endotoxin in Act-Hib vaccine, we also tried to establish the limits for the presence of endotoxin in this type of vaccine. The estimated level of endotoxin was as follows: 110 EU/ml in Act-Hib, 1.64 EU/ml in Hiberix and 2.4 EU/ml in Hib-Titer. The results of this study showed that the amounts of endotoxin was dependent on the molecular size of polysaccharide PRP and on the presence of protein component. The limit of endotoxin presence in Act-Hib vaccine recommended by us is max. 150 EU/ml.

[Physico-chemical studies of aluminum hydroxide in biological preparations preserved in different conditions]. / Ocena fizyko-chemicznego stanu wodorotlenku glinu w biopreparatach przechowywanych w róznych warunkach.

An observation of physico-chemical properties of 0.5% Al(OH)3 and vaccines adsorbed to it (Di, Te, Di-Te, Di-Te-Per) stored for 3-24 months in various pH (5, 7, 8) and temperatures -18 degrees, +37 degrees, +45 degrees and +65 degrees was carried out. For this purpose an analysis of sedimentation rate, microscopic observation and ++roentgenographic analysis were performed. It was found that a decrease of storage temperature of Al(OH)3 gel and vaccines down to -18 degrees C resulted in some changes in structure and morphology of this absorbent (big precipitates) and it led to a significant increase of sedimentation rate.

[Various immunochemical properties of nonspecific bacterial vaccines]. / Niektóre immunochemiczne wlasciwosci nieswoistych szczepionek bakteryjnych.

After critical evaluation of the composition and technology of preparation of nonspecific bacterial vaccines (nsb) some of them were withdrawn from the production (Neoflaminum, Neurovaccinum) and replaced with a new one (Polyvaccinum submite). Part of them were modified (media) and modernised (Panodinum, vaccine according to Delbet). Panodine produced at the present is free from bovine bile, which according to the results of the studies is strongly haemolytic for human and animal red blood cells. Thus it was justified to withdraw this component from Neoflaminum and Panodinum preparations. Purity and degree of lysis of these preparations were evaluated on the basis of ratio between nucleic acid absorbance and protein absorbance A260/A280. This allowed to determine if nsb tested are partially lysed and if their production process is reproducible. Our pilot studies on five patients suffering from infectious asthma indicate that a restoration of phagocytic activity of granulocytes and an increase of leukocytes migration after subsequent Panodine injections take place.

[Use of the LAL test for quantitative determination of Bacteroids fragilis endotoxin]. / Zastosowanie testu LAL do ilosciowego oznaczania endotoksyny Bacteroides fragilis.

The aim of this study was the evaluation of LAL test with chromogenic substrate usefulness for the quantitative detection of B. fragilis endotoxin and the determination of the amount of endotoxin in culture filtrates of the strains of this species. Also, the trial was undertaken to determine the influence of clindamycin on endotoxin release from B. fragilis rods to the culture medium. Four B. fragilis strains were examined: one nonenterotoxigenic (NTBF) and three enterotoxigenic (ETBF). The growth of cultures was determined and endotoxin liberated to the culture medium during growth of strains was detected. BHI broth and BHI broth with addition of sub inhibitory doses (sub-MIC) of clindamycin were applied. Bacterial cultures were incubated for 48 hours at 37 degrees C. Samples of bacterial cultures were collected after 4, 8, 16, 24 and 48 hours of cultivation, and the optical density was measured. Then the samples were centrifuged, supernatants were filtered through 0.45 micron filters and concentrated three times with 5000 D ultrafilters. Prepared samples were kept frozen at -70 degrees C until used. The amount of endotoxin in samples was determined using quantitative LAL test with chromogenic substrate S-2423. The results of the experiments indicate that LAL test is the useful method for determination of B. fragilis endotoxin concentration. This endotoxin activates the enzymatic system present in Limulus polyphemus amebocyte lysate. Endotoxin is shed spontaneously by B. fragilis rods to the culture medium during growth. Clindamycin at subinhibitory concentrations (sub-MIC) inhibits the growth of cultures of examined strains. The antibiotic caused increase in endotoxin amount in culture medium.

[LAL test used for detection of undesirable biologically active substances in biopreparations]. / Test LAL w zastosowaniu do wykrywania niepozadanych biologicznie czynnych substancji w biopreparatach.

Detection of bacterial endotoxin was performed in four groups of biopreparations: IVIG, virus and bacterial vaccines and antibiotics. The 44 samples of biopreparations were tested by the qualitative LAL-test (gel-clot) and some of them (22 samples) by the quantitative, LAL-test (chromogenic end-point). The concentration of endotoxin in 10 samples of IVIG was in the range from 0.457 EU/ml to 19.46 EU/ml. Only 4 of them did not exceed the limit recommended by FDA for human globulins (5 EU/ml). In the 9 samples out of 10 samples of virus vaccines the presence of endotoxin was in the range from 0.06 to 0.15 EU/ml. The concentration of endotoxin in 10 samples of bacterial vaccines determined by gel- clot method was below sensitivity of test (2 EU/ml). In antibiotics we did not find the presence of endotoxin in the range recommended by Ph. Eur. 1995 limits (0.1-0.2 EU/ml). The presented data show the necessity for requirements elaboration for each type of biopreparations.

[Evaluation of contamination of serum derived intravenous preparations]. / Ocena stopnia zanieczyszczenia glinem wybranych preparatów osoczopochodnych do stosowania dozylnego.

The concentration of Al in 4 lots of 5% Albumina and in 14 lots of 5% immunoglobulins were estimated by adsorptive stripping voltammetric method. Mean Al concentrations amounted to 142.6 micrograms/l in 5% Albumina and to 202 micrograms/l in IVIG preparations produced in Poland. Al concentrations in foreign IVIG preparations ranged from 14 to 54 micrograms/l. These data show the need to consider setting an upper limit of microgram Al/l in i.v. drug products produced or registered in Poland.

[The immunochemical characteristics of adult sera after Td vaccination]. / Charakterystyka immunochemiczna surowic osób doroslych szczepionych przeciw blonicy i tezcowi.

The results of immunochemical test of 29 adults sera before and after Td vaccination were presented. The level of 12 protein fractions was in normal range value in 26 of examined sera. There was IgA deficiency in one serum, in two others haptoglobin deficiency and high levels of transferrin and IgM were observed.

[Effect of selenium on peripheral blood in rats chronically exposed to benzene]. / Wplyw selenu na obraz krwi obwodowej u szczurów przewlekle eksponowanych na benzen.

The studies have been performed on 32 adult male rats of Wistar strain, divided into three groups. Control rats have been exposed to benzene vapours, of 1200 mg/m3 concentration, 6 hours daily, for 12 weeks. In the remaining two groups, before the exposure to benzene vapours, the rats had been given, to their stomachs, sodium selenate at the dose of 1.0 and 5.0 microgram/kg (converting to selenium), for 10 consecutive days. In the blood from a tail, before the experiment and after 6 and 12 weeks of exposure to benzene, some hematological indices were determined. In control group and in rats which before the exposure had been administered selenium at the dose of 1.0 microgram/kg, lymphocytopenia, correlating with duration of exposure, and thrombocytopenia were found. In rats which before the exposure had been given selenium at the dose of 5.0 microgram/kg, lymphocytopenia did not occur and thrombocytopenia occurred as late as after 12 months of exposure. The authors think that selenium at the dose of 5.0 microgram/kg, indicates a protective effect on lymphocytic system, before benzene's toxic effects.

[Heat stability of potency of bacterial vaccines and the titer of tetanus antitoxin of Polish production]. / Stabilnosc cieplna mocy szczepionek bakteryjnych oraz miano antytoksyny tezcowej produkcji krajowej.

Described the stability of potency vaccines (DTP, BCG) and immunoglobulins (human's and animal's) at storage and experimental temperatures. Thermal degradation rate and design of loss of potency in time have been determined by Arrhenius equation. Our results were similar to WHO data from preparations which have been made in another countries.

[S-II symptom questionnaire]. / Kwestionariusz objawowy S-II.

"S-II" Symptom Check-list which allows for a fast diagnosis of neurotic disorders. A result of 165 points suggests the incidence of such disorders with the probability of 90%. The methodology of the construction of the check-list intends for the application of questions most common in those ill due to neurotic disorders (owing to the change in frequency) and the most possibly equal amount of questions on the symptoms common to women and men. Thanks to this the norm for women and men is identical. SCL S-II Symptom Check-list is a shortened and actualised version of the "O" Symptom Check-list, developed in 1975. It is similar to the SCL-90 and highly correlated with it, but it does not contain the variables concerning the psychotic symptoms. Thanks to this, its' accuracy (specificity) in the diagnosis of neurotic disorders is high. 4 pairs of questions allow for the judgement of answer reliability. 10 scales were singled out in the questionnaire. They are only of a helpful value and do not allow for a one-sided diagnosis of the type of the disorder, listed in the ICD-10. The scale results can, however make the correct diagnosis easier.