[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.

[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.

[Effects of aluminum and deferoxamine on concentration of essential elements in animals experimentally intoxicated with aluminum]. / Wplyw glinu i deferoksaminy na zawartosc biopierwiastków u zwierzat doswiadczalnych poddanych intoksykacji glinem.

To assess the effect of aluminium intoxication in tissues of experimental animals on Ca, Mg, Zn, Cu, and Fe concentration, aluminium nitrate was administered intraperitoneally to mice at a daily dose of 0.27 mMol/kg for 5 weeks. Concentration of Al, Ca, Mg, Zn, Cu, and Fe were analyzed by atomic absorption spectrometry. The Al content in liver and tibia was significantly higher in treated mice in comparison with control group. In Al loaded tissues the significant increase of all tested essential elements was found. To evaluate the results of DFO treatment on essential elements, mice received 6 times intraperitoneally 3.5 mMol/kg of DFO. This treatment had generally no effect on reduction of Al concentration in tibia and liver, as well as on changes in essential elements concentration.

[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.

[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.

[Aluminum as an adjuvant in vaccines and post-vaccine reactions]. / Glin jako adiuwant w szczepionkach i reakcje poszczepienne.

Aluminium compounds have been widely used as adjuvants in prophylactic and therapeutic vaccines. Adjuvants are able to stimulate the immune system in a nonspecific manner, i.e. high antibody level can be obtained with minimal dose of the antigen and with reduced number of inoculations. Adjuvants use has been mostly empirically determined by such factors as efficacy and safety. The mechanism of action of the aluminium adjuvants is not completely understood and is very complex. The basic factors of the mode of action: 1) the complex of antigen and aluminium gel is more immunogenic in structure than free antigen, 2) effect "depot"--The antigen stimulus last longer, 3) the production of local granulomas. Vaccines adsorbed onto aluminium salts are a more frequent cause of local post-vaccinal reactions than plain vaccines. 5-10% those vaccinated can develop a nodule lasting several weeks at the injection site. In some rare cases the nodules may become inflammatory and even turn into an aseptic abscess. The nodules persisting more than 6 weeks may indicate development of aluminium hypersensitivity. Finally aluminium adjuvant immunogens induce the production of IgE antibodies.

The level of endotoxin contamination in biopreparations.

Bacterial endotoxins as contamination of biopreparations have been estimated by chromogenic LAL test. Study on some compounds (aluminium hydroxide, formaldehyde and merthiolate) being components of vaccines showed no effect on the result of LAL test. The level of endotoxins in virus vaccines with the limits defined in producers certificate was adequate, the level of endotoxin was also low in virus vaccines of undefined requirements. The concentration of endotoxin in bacterial vaccines was differentiated. Considering the results of our experiments, as well as the fact, that the requirements for endotoxin contamination of bacterial vaccines are not available it seems necessary to establish the limits for these group of biopreparations.

Effect of aluminium hydroxide administration on normal mice: tissue distribution and ultrastructural localization of aluminium in liver.

In order to assess the risk of parenteral aluminium (Al) exposure, we evaluated the effects of intraperitoneal administration of aluminium hydroxide, a compound widely used in medicine. Mice (strain Pzh:SFIS) received intraperitoneally, every two weeks 1 mg Al or 0.1 mg Al for five days a week. Controls received injections of saline. Al concentrations in liver, bone and brain were evaluated by electrothermal atomic absorption spectrometry after exposure to 2 mg, 4 mg, and 6 mg Al. The concentration was the highest in liver and occurred after exposure to only 2 mg Al (265.1 +/- 27.7 mg/kg, 233.5 +/- 28.0 mg/kg). Generally further accumulation was not dose- and treatment-dependent. The only exception was a significant Al increase in the liver after exposure to 6 mg Al, injected 0.1 mg Al five days/week. Development of resorption granulomas was observed in the liver, Al being revealed by Morin fluorescence in constituent macrophages and giant cells. By electron probe X-ray microanalysis, Al was identified predominantly in lysosomes of macrophages and Kupffer cells. In tibia of mice, a dose-dependent Al accumulation was observed. The highest level of Al concentration after the 6 mg treatment was 23.5 +/- 3.82 mg/kg and 25.06 +/- 2.3 mg/kg. The Al concentration in the brain of mice had not changed significantly during Al treatment.

Effects of various aluminium compounds given orally to mice on Al tissue distribution and tissue concentrations of essential elements.

To evaluate the risk of gastrointestinal long-term aluminium (Al) exposure, aluminium distribution and the levels of the following essential elements: Ca, Mg, Zn, Cu, and Fe in tissue were studied. Aluminium was administered in drinking water as aluminium chloride, dihydroxyaluminium sodium carbonate or aluminium hydroxide. Mice (strain Pzh:SFIS) were exposed to a total dose of 700 mg Al in long-term treatment (for each Al compound n = 15). Concentrations of Al, Ca, Mg, Zn, Cu, and Fe in stomach, kidneys, bone and liver were analyzed by atomic absorption spectrometry. After AlCl3 treatment, aluminium was found to accumulate in all tested tissues. A significant decrease in Fe concentration in liver and Zn in kidneys was observed in comparison to concentrations of these elements in the control group. In the Al(OH)3-treated group, accumulation of aluminium was observed in bone only and decline of Fe concentration in stomach and Cu in liver and kidney. In the NaAl(OH)2CO3-treated group the increase in Al concentration was significant in bone; there was no change in concentration of essential elements in the examined tissues. The observed aluminium accumulation was not accompanied by changes in Ca and Mg concentration except for bone. This study showed that oral administration as a route of Al exposure can result in diverging accumulation of aluminium in tissues, the concentration depending on the chemical form.