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[Biological assay, reliability test, error analysis and pooled calculation for assessing leucomycin yield].

A biological assay for determining leucomycin yield was established by using a curillinear protocol (Sarcina lutea) and (2.2) dosage or (3.3) dosage protocol (Bacillus subtillis 63501). At the same time, according to the principles of biostatistics and the methods of variance analysis, reliability tests, error analysis and pooled calculations were carried out, so as to hold the confidence limit below the internationally accepted level of 5%.

Biological Assay↗

[Shigella endotoxin protein--its electrophoretic and serological properties].

The electrophoretic analysis of lipid A-associated protein (LAP), obtained from S. sonnei, in polyacrylamide gel in the presence of sodium dodecyl sulfate and urea has revealed the heterogeneity of the preparation; it has found to contain three main components with molecular weights of 43, 38 and 18 KD and some minor components with molecular weights of 49, 45 35, 30, 29, 27, 5, 21 and 14 KD. The electrophoretic mobility of the main protein components in the isolated preparation of LAP coincides with that of endotoxin components. The dissociation of proteins and lipopolysaccharide in the process of boiling the endotoxin in 2% sodium dodecyl sulfate is indicative of the noncovalent binding of these components. LAP contained in the endotoxin, in contrast to isolated LAP, is resistant to trypsin and proteinase K. The enzyme immunoassay (EIA) system with the use of LAP as a component of its solid phase has been developed, which makes it possible to carry out the quantitative determination of antibodies to this protein. The EIA system shows high sensitivity in the determination of anti-LAP IgG antibodies: in hyperimmune rabbit sera their titer is 1:250,000-1:800,000. As shown by the method of competitive EIA, the antigenic affinity of LAP of different origin corresponds to the degree of taxonomic propinquity of microorganisms: the maximal degree of cross reactions is observed between LAP obtained from S. sonnei, S. flexneri and Escherichia coli, while their affinity to Salmonella typhi is considerably less; remote microbial species (Bacterium bifidum and Sarcina marcescens) give practically no cross reactions.

Animals↗

Synthesis and some biological properties of 6-aminopenicillanic derivatives.

The properties of 33 6-aminopenicillanic derivates were described, the majority of which and particularly all 3,4-disubstituted phenyl derivatives, are new compounds. These derivatives were synthesized by three different methods consisting in treating of 6-aminopenicillanic acid (6-APA) with appropriate phenylthio -- and phenoxyalkanecarboxylic acids. The method consisting in use of aryloxyketenes is new and claimed by the patent applications. All synthesized compounds were screened for their biological activity against 21 different strains of pathogenic microorganisms of species: Staphylococcus, Sarcina, Streptococcus, Escherichia, Proteus, Klebsiella, Neisseria, Pseudomonas, Bacillus subtilis and Candida albicans. In the present paper only the most interesting results of microbiological tests and some data concerning pharmacological investigations performed on rats, were presented. Among selected compounds both substances 30 and 32 showed animals weight growth stimulating activity.

Animals↗

[The aerobic air microflora in airplanes on various international routes].

Aerobic microflora (bacteria, fungi), in the cock pits of the TAROM company (Boeing 707 and Il 62 M) airships flying on various international routes and airports was studied during November 1988-January 1989. 157-8,800 bacteria and 78-1,336 fungi per m3 air were recorded. Except for Staphylococcus aureus (hemolytic and non hemolytic) the greatest part of the isolated microorganisms was nonpathogenic for man: Bacillus, Corynebacterium, Neisseria, Staphylococcus epidermidis, Sarcina, Aspergillus, Penicillium etc. Several airships on the Asian airports contained a higher amount of bacteria and fungi but not higher than in the living rooms. Likewise, in high altitude flights, the microorganism amount was less than on the ground. The taxonomic spectrum of the bacteria and fungi isolated was almost identical on all the 9 international airports, thus suggesting the homogeneous and international character of saprophyte and pathogenic air microflora by means of the passenger and goods air flights.

Air Microbiology↗

[Fundamental and clinical investigation of clindamycin-2-phosphate in otorhinolaryngological field (author's transl)].

Some laboratory examinations were made with clindamycin-2-phosphate (CLDM-2-phos.). The drug was applied clinically to several otorhinolaryngological infections and the good results were obtained as follows. 1) Observing by biophotometer, the growth of Staphbylococcus aureus FDA 209P was inhibited well by the serum (diluted 10-fold) 30 minutes, 1, 2, 4 hours after 300 mg of CLDM-2-PHOS. were injected intramuscularly. 2) Serum concentration was determined by thin-layer method with Sarcina lutea ATCC 9341 as the test organism. The peak of serum level was obtained at 1 hour after a single intramuscular injection of 5 mg/kg or 10 mg/kg dosis of CLDM-2-phosphate. The peak of blood concentration was 5.4 mug/ml, 5.6 mug/ml respectively. 3) CLDM-2-phosphate concentration in tissues was estimated one hour after intramuscular injection of CLDM-2-Phosphate. The concentrations in blood and palatine tonsilla were 5.0 mug/ml and 2.9 mug/ml respectively after the intramuscular injection of 10 mg/kg. The concentrations in blood and mucous membrane of maxillar sinusitis were 4.1 mug/ml and 2.5 mug/ml respectively after the intramuscular injection of 10 mg/kg. 4) CLDM-2-phosphate was administered by intramuscular injection into 30 cases with various infections in otorhinolaryngological field. The clinical results were excellent in 22 cases (73.3%), effective in 3 cases (10.0%) and ineffective in 5 cases (16.7%). The effectiveness was 83.3%. 5) No side effects were observed in 30 cases.

Acute Disease↗

Antimicrobial activity of julifloricine isolated from Prosopis juliflora.

Antimicrobial activity of julifloricine, an alkaloid isolated from Prosopis juliflora, was studied in vitro against 40 microorganisms which included 31 bacteria, two Candida species, five dermatophytic fungi and two viruses. Significant inhibitory effect was noted against Gram positive bacteria. The minimal inhibitory concentration (MIC) for Staphylococcus aureus, S. epidermidis, S. citreus, Streptococcus pyogenes and Sarcina lutea was 1 microgram/ml and against S. faecalis, S. pneumoniae, S. lactis, Corynebacterium diphtheriae, C. hofmannii and Bacillus subtilis, 5 micrograms/ml. Its effect was compared with those of identical concentrations of benzyl penicillin, gentamicin and trimethoprim. The inhibitory effect of julifloricine on Gram negative bacteria such as the species of Salmonella, Shigella, Klebsiella, Proteus, Pseudomonas, Enterobacter, Aeromonas and Vibrio was almost insignificant. Julifloricine as compared to micoanzole was found superior against C. tropicalis and responded equally to C. albicans. As compared to econazole, it was found less effective against both C. albicans and C. tropicalis. This alkaloid was found inactive against dermatophytic fungi (up to a dose of 10 micrograms/ml) and viruses which included Herpes simplex 1 and Newcastle disease virus. Julifloricine up to a doses of 1000 micrograms/25 g of mice was found nonlethal.

Alkaloids↗

Food-borne amines and amides as potential precursors of endogenous carcinogens.

This paper reviews the experimental results of our research in the past several years and other related papers that have been directed toward the occurrence, biotransformation and epidemiological significance of carcinogenic N-nitroso compounds in biosphere. Endogenous carcinogens are a group of cancer-causing compounds produced in vivo from harmless precursors. This category has been exemplified by the well-known carcinogens, N-nitroso compounds. The significance of naturally occurring amines and amides as precursors of carcinogenic N-nitroso compounds in vivo and their implication in the incidence of human cancer have been investigated and emphasized. Extremely high levels of trimethylamine-N-oxide and dimethylamine were detected in squids and other seafoods. More than 90% of trimethylamine-N-oxide were converted to dimethylamine and trimethylamine on pyrolysis. Low levels of dimethylamine and methylamine were also detected in the fermented soybean products, wines and sauces. Both dimethylamine and trimethylamine are excellent precursors of dimethylnitrosamine. Several naturally occurring aromatic amines especially 2-carboline derivatives such as harman, norharman, harmaline, harmalol, harmine and harmol are mutagenic and become more mutagenic to Salmonella typhimurium after nitrosation. Appreciable amounts of piperidine were detected in the popular spice white and black pepper powders. Under acidic condition, piperidine reacts readily with nitrite to form carcinogenic N-nitroso-piperidine. N-Nitrosophenacetin was formed from the reaction of nitrite with the amide drug phenacetin. This new compound showed strong mutagenicity to Salmonella typhimurium and Sarcina lutea and strong teratogenic activity to Leghorn chicken embryos. Studies have shown that the majority of N-nitroso compounds in the body come from in vivo conversion. Most investigators believe that this endogenous pool of N-nitroso compounds may prove to be a major exposure route in man. The presence of naturally occurring amines and amides in the diet then becomes one of the crucial limiting steps in the formation of endogenous N-nitroso compounds in vivo.

Amides↗

Plasma penicillin-G parameters in man after intraduodenal or intramuscular administration.

Concentrations of penicillin-G produced in serum by either direct intraduodenal (i.d.) or intramuscular (i.m.) administration were compared in 9 healthy volunteers. Serum specimens were assayed by the Sarcina lutea cup plate method. The peak serum levels achieved by i.d. application were similar to those observed after intramuscular dose. However, higher concentrations were better sustained after i.m. penicillin. We concluded that intraduodenal administration of penicillin-G could be considered as an alternative for i.m. injection.

Absorption↗

Comparison of liquid chromatographic and bioassay procedures for determining depletion of intramuscularly injected tylosin.

Crossbred pigs weighing 80-110 kg were injected intramuscularly in the ham with 8.8 mg/kg tylosin. Animals were slaughtered in groups of 3 at intervals of 4 h, and 1, 2, 4, and 8 days after injection, and samples of blood, injected muscle, uninjected muscle, liver, and kidney were analyzed by liquid chromatography (LC) and by bioassay using Sarcina lutea as the test organism. The LC method was far more sensitive with a detection limit of less than 0.1 ppm, while the detection limit by bioassay was about 0.5 ppm in tissue. Results by bioassay and LC sometimes differed considerably for tissue samples. Residues in all tissues were below the tolerance limit of 0.2 ppm at 24 h, except in the injected muscle in one animal. Residues were not detected in any tissue of any animal at 48 h after treatment.

Animals↗

[Antimicrobial activity of some medicinal species from the Dakar markets].

After drawing up an inventory of Dakar markets popular medicinals plants, antimicrobial activity of 43 species have been screened. With the agar diffusion method, crude extracts of 24 species show an activity against Sarcina lutea or Escherichia coli. The antimicrobial spectrum of these extracts against 11 germs has been evaluated with the agar dilution method. 13 species show M.I.C. lower than 10 mg/ml against some germs. This preliminary study select some plants of potential interest for development of new antibacterial.

Anti-Bacterial Agents↗

Connections between Ascaridia galli and the bacterial flora in the intestine of hens.

Parasitological dissections of 502 intestinal tracts of hens deriving from big private chicken-farms have been done. In the jejunum of 146 hosts (ext. 29.1%) from 1 to 21 individuals of A. galli were detected. Using bacterial selective media and biochemical tests, the microorganisms from the hen's intestinal tracts as well as from the cuticle surface of the nematodes were identified. Among them were: grampositive (+) Lactobacillus, Bacillus, Staphylococcus, Streptococcus, Micrococcus, Sarcina, Clostridium, Corynebacterium; gramnegative (-) Enterobacteriaceae, Pseudomonas, Pasteurella, and fungi Candida and others. The lower frequency of microorganisms and the smaller amount of bacteria in the intestinal content in infected hens than in uninfected show that A. galli has antibacterial properties.

Animals↗

Bacillus stearothermophilus disk assay for detection of residual penicillins in milk: collaborative study.

A collaborative study was performed on a Bacillus stearothermophilus paper disk method designed to detect residual levels of 4 antibiotic drugs in whole market milk. This method is a modification of an earlier procedure developed for the International Dairy Federation. Whole milk samples spiked at low levels with ampicillin, cephapirin, cloxacillin, and penicillin G were sent frozen to 11 collaborating laboratories with instructions to assay them promptly according to the method provided. Five of the laboratories reported inconclusive results due to technical difficulties encountered with the method. The 6 remaining laboratories all detected levels of 0.005-0.008 microgram or unit/mL for penicillin G, ampicillin, and cephapirin and 0.05-0.08 microgram/mL for cloxacillin. The most commonly used official methods, the Sarcina lutea (Micrococcus luteus) cylinder plate method and the Bacillus subtilis paper disk method, can detect levels of 0.01 and 0.05 unit penicillin G/mL, respectively. The B. stearothermophilus method is rapid, simple to perform, and more sensitive than present official methods. The method has been adopted as official first action for the detection of penicillins in milk.

Animals↗

N-nitrosophenacetin: its synthesis, characterization, mutagenicity, and teratogenicity.

Reaction of phenacetin (CAS: 62-44-2; p-acetophenetidide) with nitrous fumes (N2O3) in glacial acetic acid at 0-5 degrees C yields N-nitrosophenacetin (NP), 2-nitrophenacetin, N-nitroso-2-nitrophenacetin (NNP), and other compounds. Both NP and NNP are fairly stable at low temperature (-30 degrees C) but extremely labile at ambient temperature. NP (median lethal dose to Sprague-Dawley rat: 21 mg/kg body wt) is 80 times more toxic than its parent compound phenacetin and is directly mutagenic to bacterial cells including Salmonella typhimurium and Sarcina lutea. The mutagenicity of NP is comparable to that of N-methyl-N'-nitro-N-nitrosoguanidine [(MNNG) CAS: 70-25-7; 1-methyl-3-nitro-1-nitrosoguanidine] and requires no microsomal metabolic activation. The teratogenic potential of NP was studied in White Leghorn chick embryos given a single dose of 5-15 micrograms/egg on day 6 of incubation. A low incidence of exencephaly and eyelid defect and a high incidence of feather and claw malformations were found in the treated group; no such malformed embryos were found in the control group. The teratogenicity of NP was found to be weaker than that of MNNG, but stronger than that of N-methyl-N-nitrosourea (CAS: 684-93-5), dimethylnitrosamine (CAS: 62-75-9; N-nitrosodimethylamine), and diethylnitrosamine (CAS: 15-18-5; N-nitrosodiethylamine).

Abnormalities, Drug-Induced↗

[Airborne microflora found in some stations of the metro in the Hungarian capital of Budapest (author's transl)].

Both the microflora and some physical parameters of the air in three underground railway (metro) stations were analysed. The air samples taken in the first two stations contained considerably more microbes, namely three times as many, than the sample from station No. 3. In the samples from the first two stations both the pathogenic and anthropogenic germs occured more frequently than in the samples from the third station. The normal flora consisted of gram-positive cocci/Streptococci, Micrococci, Sarcina. Roughly 10 per cent of the microbes proved to be spore-formers. Selective investigations led to the detection of Staph. aureus, Enterococci, Haemophilus influenzae, Escherichiae, Klebsiella and Proteus bacteria. The reason why the stations of line 2 were found to be contaminated to a greater degree is probably because of the higher passenger traffic, the smaller space, as well as the stronger airflow due to the artificial ventilation and the piston effect of the trains. The effect of the artificial environmental conditions on the staff of the metro company still needs further investigation. The effect on the passengers is considered to be less pronounced.

Air Microbiology↗

Determination of penicillin G, ampicillin, and cephapirin residues in tissues.

A cylinder plate microbiological method was developed for the rapid, quantitative determination of penicillin G, ampicillin, and cephapirin in animal tissues. The method uses agar plates seeded with stable spores of Bacillus stearothermophilus var. calidolactis and incubated 4 h at 64 degrees C. Standard curves were obtained for the following ranges of concentration of antibiotics in tissues: 0.02-0.32 IU penicillin G/g, 0.0125-0.2 micrograms ampicillin/g, and 0.02-0.32 micrograms cephapirin/g. The proposed method is suitable not only for penicillin residue analysis, for which the sensitivity has been greatly improved compared with the Sarcina lutea method, but also for depletion studies on these antibiotics, which are commonly used to treat diseases in food-producing animals.

Ampicillin↗

Oral penicillin prophylaxis in children with imparied splenic function: a study of compliance.

One measure used to prevent overwhelming sepsis due to Streptococcus pneumoniae in children with defective splenic function is oral penicillin prophylaxis. However, a frequently cited argument against this approach is the likelihood of poor compliance. Compliance was studied by examining urine specimens for penicillin by the Sarcina lutea disc diffusion technique in 22 surgically asplenic children, two patients following bone marrow transplantation, and 38 infants and young children with sickle cell disease. Multiple specimens (mean 3.5 per patient) were examined in 43 of the children. Overall, 125/188 (66%) of the urine samples contained penicillin, indicating compliance within the previous 12 to 24 hours. Compliance tended to improve on subsequent clinic visits. These relatively good results were attributed to an intensive educational program in which repetitive efforts are made to counsel patients and parents about the risks of life-threatening infection. Poor compliance should no longer be invoked as a reason not to study the efficacy of prophylactic penicillin in functionally or surgically asplenic subjects.

Administration, Oral↗

[Preservation of lyophilized cultures of saprophytic microorganisms].

All of the tested saprophytic microbial cultures remained viable upon 11--16 years of storage in the lyophilized state at 3--6 degrees. The cultures belonged to the following taxonomic groups: Acetobacter, Achromobacter, Azotobacter, Bacillus, Chromatium, Escherichia, Flavobacterium, Lactobacillus, Micrococcus, Proteus, Propionibacterium, Pseudomonas, Rhizobium, Sarcina, Serratia, Streptococcus, Torula, Mycobacterium, Actinomyces. The number of viable cells in the tubes decredased only slightly within this time and remained at the same level as immediately after lyophilization.

Bacterial Physiological Phenomena↗