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[The significance of heat activation for the testing of bioindicators on surviving microorganisms exposed to formaldehyde].

Heat activation is a special phenomenon: After an additional heat treatment, a larger share of the bacterial spores which had been exposed to formaldehyde proves to be viable than without such heat activation. Model studies have been performed to test the effects of heat activation on the examination of bioindicators and test objects for surviving organisms. Test objects (cotton threads of 1 cm length) contaminated with spores of Bacillus stearothermophilus were used for these trials. The test objects were exposed to a 2% formaldehyde solution at 60 degrees C. After periods of action of 30, 45, 60 ... and 105 min, formaldehyde adhering to the test objects was neutralized. For testing these objects for surviving organisms, they were placed into a nutrient medium and incubated for 40 days at 56 degrees C. The investigation consisted of 2 parallel test series which only differed in one single point. In one series, the test objects were incubated at 56 degrees C as soon as they had been placed into the nutrient solution. In the other series, the test objects were exposed to a temperature of 95 degrees C for 1 h (heat activation) before starting incubation. The culture tubes were checked daily to see whether signs of growth (turbidity and deposits) could be observed. The frequencies of test objects with surviving organisms depending on the period of action of formaldehyde and the period of incubation determined in this way are based on the examination of 72 test objects each. Without heat activation, the share of test objects on which surviving test organisms could be detected, increased slowly with the period of incubation. Only after 30 days the counts did not increase any more when continuing the incubation (cf. Fig. 1). In the test series in which the spores had been subjected to heat activation before the incubation period, useful results were obtained already after 3 days. They only changed slightly when incubation was continued. Moreover, the frequency of test objects on which surviving organisms could be detected was always considerably higher than without heat activation. When the frequency of test objects with surviving organisms was plotted against the period of action of formaldehyde (cf. Fig. 2A), S-shaped curves resulted.(ABSTRACT TRUNCATED AT 400 WORDS)

Formaldehyde↗

Effects of formaldehyde on the mucous membranes and lungs. A study of an industrial population.

One hundred nine workers and 254 control subjects were studied to evaluate the effects of formaldehyde on the mucous membranes and lungs. A modified, respiratory symptom questionnaire and spirometry were administered to all study participants before and after their work shift, and formaldehyde levels were determined for each test subject. Over the course of the monitored work shift, test subjects demonstrated a dose-dependent excess of irritant symptoms and a statistically significant decline in certain lung function parameters. Analysis of test and control subject data combined revealed a correlation between formaldehyde exposure and these pulmonary changes. Baseline spirometry values were not significantly different between test and control groups, and formaldehyde-exposed workers did not report an excess of respiratory symptoms. Formaldehyde is a dose-dependent irritant of the eyes and mucous membranes at low-level exposures. It can exert a small, across-shift effect on airways but after a mean exposure of ten years does not appear to cause permanent respiratory impairment.

Adult↗

Cytologic examination of the nasal mucosa in formaldehyde-exposed workers.

A cross-sectional study of 80 individuals at three locations was undertaken to evaluate the health effects of long-term exposure to formaldehyde in a phenol-formaldehyde impregnating process and to develop a useful protocol for health surveillance of formaldehyde-exposed workers. Results of physical examinations showed a statistically significant prevalence of mucosal irritation in formaldehyde-exposed workers, particularly those with recent exposure. Cytologic examination of exfoliated nasal cells showed atypical squamous metaplasia, which was found to be a function of age. There was no statistical relationship to formaldehyde exposure.

Adult↗

[Permeability of the blood-brain barrier to intra-arterial formaldehyde].

Formaldehyde concentration was assessed in the brain, cerebrospinal liquor, arterial and venous blood of intact animals and following its intraarterial injections. It is concluded that formaldehyde is capable of penetrating through the blood-brain barrier, with the degree of permeability depending on blood formaldehyde concentration. The distribution of formaldehyde in the blood-brain-cerebrospinal liquor system suggests the presence of both protein-bound and unbound formaldehyde forms in the organism.

Animals↗

[Changes in the sorption properties of LDH protein in the rat brain after the single administration of a formaldehyde solution].

The study of the activity, protein charge and isoenzyme LDG spectrum after a single injection of formaldehyde solution has demonstrated a rise in the protein negative charge, which in the absence of ischemia was observed for 20 min. A more than 5-min increase in LDG protein charge observed in ischemia and independent of changes in LDG isoenzyme spectrum was significantly less expressed in ischemic brain after a preliminary injection of formaldehyde solution. With cerebral blood flow preserved, an injection of formaldehyde resulted in the alteration of LDG spectrum, namely a rise in LDG3+4+5 content. Formaldehyde injection prior to ischemia prevented the development of a similar effect, which was significantly more expressed in ischemic brain without a preliminary formaldehyde injection.

Absorption↗

Some aspects of residual formaldehyde testing when reusing haemodialysers.

To assess the adequacy of quantifying residual formaldehyde concentrations when reusing, four semi quantitative methods of concentration estimation (Clinitest tablets, Schiff's reagent, Formalert Formotest) were compared. Two of these methods (Schiff's reagent and Clinitest) were inadequate in detecting low concentrations of formaldehyde and were associated with false positives from interference by chemicals contained in the dialysate. When using the two reagents specifically manufactured for this purpose, we also observed false positives with one (Formotest) while the other was capable of detecting formaldehyde concentrations as low as 4.5 mg/l. To minimise patients' exposure and the formation of anti-N like antibodies resulting from formaldehyde damage of red cells, it would be prudent when reusing, to select the most sensitive of these semi-quantitative techniques for routine use, to perform regular screening for anti-N antibodies and to periodically check formaldehyde levels by the use of the highly specific Hantzsch reaction.

Formaldehyde↗

[Formaldehyde-induced tight binding of protein to RNA in particles of rod-like virus].

The formaldehyde-induced formation of tightly bound RNA-protein complexes of rod-like plant viruses was studied. The preparations of tobacco mosaic virus and closely related cucumber virus 4 were incubated with 1.5% formaldehyde for 20-50 hrs at 50 degrees C. Then the viral particles were disrupted, free protein was removed and viral RNA was centrifuged in the linear gradient of Cs2SO4. The RNAs from the formaldehyde-untreated viruses and RNA from the formaldehyde-treated tobacco masaic virus had the density of 1.65-1.66 g/cm3, while RNA from the formaldehyde-treated cucumber virus had the density of 1.57-1.42 g/cm3, depending on the incubation time. This is indicative of the protein binding to RNA. Treatment of the cucumber virus complex with pronase resulted in a liberation of free RNA with the density of 1.66 g/cm3; incubation for 2 min at 100 degrees C in a dissociating mixture (2% sodium dodecyl sulfate + 0.2% mercaptoethanol) did not cause the dissociation of the complex. Polyacrylamide gel electrophoresis showed that the most part of the protein molecules are bound within the complex not by covalent protein-protein cross-links.

Formaldehyde↗

[The action of products of the formaldehyde reaction with different amines on nucleic acids and their components].

The kinetics and equilibrium of the reaction between nucleic acids components and the products of formaldehyde interaction with ethanolamine and different amino acids has been studied. These parameters were found to be similar for all the products used. The destabilization of the N-glycosidic bond in deoxyadenosine caused by formaldehyde derivatives of different amines was studied. The rate of the cleavage of the N-glycosidic bond under the action of formaldehyde derivatives of glycine and ethanolamine was found to be 10 times greater than that under the action of formaldehyde derivatives of other amines. It is shown that DNA preparations with different content of adenine can be obtained by adding the product of formaldehyde reaction with glycine to DNA.

Amino Acids↗

Formaldehyde dehydrogenase from Pseudomonas putida. Purification and some properties.

Formaldehyde dehydrogenase was isolated and purified in an overall yield of 12% from cell-free extract of Pseudomonas putida C-83 by chromatographies on columns of DEAE-cellulose, DEAE-Sephadex A-50, and hydroxyapatite. The purified enzyme was homogeneous as judged by disc gel electrophoresis and was most active at pH 7.8 using formaldehyde as a substrate. The enzyme was also active toward acetaldehyde, propionaldehyde, glyoxal, and pyruvaldehyde, though the reaction rates were low. The enzyme was NAD+-linked but did not require the external addition of glutathione, in contrast with the usual formaldehyde dehydrogenase from liver mitochondria, baker's yeast, and some bacteria. The enzyme was markedly inhibited by Ni2+, Pd2+, Hg2+, p-chloromercuribenzoate, and phenylmethanesulfonyl fluoride. The molecular weight of the enzyme was estimated to be 150,000 by the gel filtration method, and analysis by SDS-polyacrylamide gel electrophoresis indicated that the enzyme was composed of two subunit monomers. Kinetic analysis gave Km values of 67 microM for formaldehyde and 56 microM for NAD+, and suggested that the reaction proceeds by a "Ping-pong" mechanism. The enzyme catalyzed the oxidation of formaldehyde accompanied by the stoichiometric reduction of NAD+, but no reverse reaction was observed.

Aldehyde Oxidoreductases↗

[Estimation of various quantitative methods for the determination of native and formaldehyde-treated proteins].

Quantitative estimation of native and treated with 4% formaldehyde albumin, pepsin, lysozyme, histone, gelatin and other proteins was carried out using four procedures--biurete, Lowry-Folin, ninhydrin and coomassy R-250. Chromogeneity of proteins in corresponding color reactions was expressed as OD per 100 micrograms of nitrogen estimated by means of Kjeldahl micromethod. The proteins were treated with formaldehyde in corresponding buffers at 20 degrees within 7 days, non-bound or loosely bound formaldehyde was dialysed. Native proteins were dissimilar in their chromogeneity; these differences were the highest for Lowry-Folin and coomassy procedures. Formalinization affected the protein chromogenencity depending on a protein nature, conditions of formaldehyde treatment and on the procedure of estimation used. As shown by analysis of the data obtained the alterations of chromogeneity did not reflect the rate of reactive group blocking by formaldehyde in a protein molecule. The quantitative methods should be used very carefully in estimation of formalinized proteins.

Albumins↗

[Disinfection and sterilization of thermolabile instruments with gaseous formaldehyde].

For several years formaldehyde-gas-underpressure procedures are increasingly used for disinfection and sterilization of medical thermolabile instruments. In many places, however, uncertainty and sceptism, if these methods are sufficient in the medical field, are existing. This is understandable, since no generally test instructions are available. The principal suitability of gaseous formaldehyde for disinfection and sterilization, however, had been demonstrated by several investigators. Precondition for reliable efficacy during routine use is an apparatus which is equipped with feed-back techniques and which guarantees the bactericidal and fungicidal activity required for medical use. Decontamination procedures on the basis of gaseous formaldehyde under normal conditions of temperature and pressure must be regarded as unsafe. St. faecalis and the spores of B. stearothermophilus showed the highest formaldehyde resistance within the group of common biological testorganisms. In order to test the efficacy of corresponding procedures testorganisms have to be placed into testdevices with small lumina. The penetration of these devices to formaldehyde and vapour should be similar to that of the longest instruments with the smallest lumina, which are intended to be decontaminated by the testprocedure.

Bacillus subtilis↗

Use of formaldehyde-PBS for serum dilution in the microscopic agglutination test (MAT) for leptospirosis.

Altogether, 431 sera (381 positive and 50 negative sera) were tested against different Leptospira serovars in the microscopic agglutination test (CMAT) using PBS with and without formaldehyde for comparative purposes. For the preparation of serum dilutions with formaldehyde-PBS, formaldehyde was added to PBS at a final concentration of 0.4%. When retested after storage, 234 from the 381 formerly positive sera gave positive, 68 doubtful and 79 negative results in the MAT. Out of the 234 sera with positive reaction in MAT, 212 (90.6%) showed positive reactions in the MAT with formaldehyde as well, 19 (8.1%) doubtful reactions and 3 (1.3%) became negative. All sera with negative reaction in the routine MAT were found to be negative in the MAT with formaldehyde as well.

Agglutination Tests↗

[Characteristics of formaldehyde reactions with nucleic acids and their structural components in the presence of primary or secondary amines].

Peculiarities of reversible interaction of formaldehyde with DNA, adenine and some cytosine derivatives in the presence of primary or secondary amines have been studied. Different characters of the examined formaldehyde reactions have been demonstrated. Thus, melting of DNA by formaldehyde at neutral pH is accelerated only in the presence of primary amines. Using UV-spectra data analysis it was concluded that products of cytidine and adenine modification by formaldehyde or methylol derivatives of secondary amines have similar structure. Cytidine and adenine modifications when treated with formaldehyde and primary amine mixture were considerably different and resulted in products with different structures. This is probably related to interactions within two positions of cytosine or adenine residues. A hypothetical scheme of formation and structure of modification products are considered.

Adenine↗

[Mechanism of the cytotoxic action of formaldehyde on cultured mammalian cells].

Induction and repair of DNA lesions cell inactivation and repair of potentially lethal damages (PLD) were studied after the treatment of cultured cells with formaldehyde. Formaldehyde induced the appearance of a rapidly sedimentating DNA--membrane complex. This complex may contain up to 50% of choline and no more than 3-5% of leucine or lysine incorporated in the acid insoluble cell fraction, Inhibition of DNA synthesis, induction of single strand DNA breaks and/or alkali-labile sites increased with the raise of formaldehyde concentration. A good correlation is observed between with the raise of formaldehyde concentration. A good correlation is observed between with the raise of formaldehyde concentration. A good correlation is observed between the increasing DNA quantities in the rapid sedimentation complex and the cell lethality.

Animals↗

[Formaldehyde sediment in incubators following disinfection].

Measurements in incubators revealed the presence of formaldehyde concentrations involving a health risk for premature and normal newborns kept and cared for in incubators. Prior to measurements, the incubators had been disinfected by means of formaldehyde vapours in an "Aseptor" disinfecting cabinet (Drägerwerk AG, Lübeck) and then ventilated in strict adherence to operating instructions. The elevated formaldehyde concentrations found had been due to residues of paraformaldehyde and urotropin on the surfaces of the disinfected apparatus, liberating formaldehyde by hydrolysis depending on temperature and relative humidity. There should be a basic reconsideration of the present practice of incubator disinfection. From experiments with activated-carbon filters in incubators it would seem that there is a chance of reducing such formaldehyde concentrations.

Disinfection↗

[Kinetics of formaldehyde splitting off from the hydroxymethylated amino groups of nitrogen bases incorporated into double-helical DNA].

We studied the kinetics of formaldehyde dissociation from hydroxymethylated amino groups of nitrous bases on native DNA. Compared with monomers, the rate constant of formaldehyde dissociation from such bases integrate in the double helix proved to be 20 times smaller for adenine and 4 times smaller for cytidine within the temperature range of 15 to 40 degrees C. The kinetic pattern suggests that the dissociation of formaldehyde from hydroxymethylated amino groups does not occur in the direction of the base plane nor through a full fluctuational opening of base pairs. It is presumed that formaldehyde dissociation from modified amino groups is due to softer fluctuational changes which however, make it possible for formaldehyde to attack amino groups perpendicularly to the base plane.

Chemical Phenomena↗

Induction of squamous cell carcinomas of the rat nasal cavity by inhalation exposure to formaldehyde vapor.

Groups of 120 male and 120 female rats were exposed by inhalation to 0, 2, 6, or 15 ppm formaldehyde vapor 6 hr/day, 5 days/week, for 18 months of a 24-month study. The present communication describes interim findings based on data available after 18 months of exposure. Squamous cell carcinomas occurred in the nasal cavities of 36 rats exposed to 15 ppm formaldehyde. The tumors ranged from small early carcinomas of the nasal turbinate to large invasive osteolytic neoplasms which extended into the subcutis of the premaxilla. Similar tumors were not detected in rats exposed for 18 months to 2 or 6 ppm or in mice exposed to 2, 6, or 15 ppm formaldehyde. Rhinitis, epithelial dysplasia, and squamous metaplasia occurred in rats from all exposure levels of formaldehyde; however, the severity and extent of the lesions were dose related. In contrast, papillary hyperplasia and squamous atypia occurred only in animals exposed to 15 ppm formaldehyde.

Animals↗

Cytogenetic effects of formaldehyde exposure in students of mortuary science.

The effect of low-level exposure to formaldehyde on oral, nasal, and lymphoycte biological markers was studied prospectively in a group of 29 mortician students who were about to take a course in embalming. During the 85-day study period, the subjects performed an average of 6.9 embalmings and had average cumulative formaldehyde exposures of 14.8 ppm-h, with an average air concentration of 1.4 ppm during embalming. Since the average time spent embalming was 125 min, formaldehyde exposures calculated as an 8-h time-weighted average were 0.33 ppm on days when embalmings were done, which was less than the Occupational Safety and Health Administration permissible exposure limit of 0.75 ppm. Epithelial cells from the buccal area of the mouth showed a 12-fold increase in micronucleus frequency during the study period, from 0.046 +/- 0.17/1000 cells preexposure to 0.60 +/- 1.27/1000 cells at the end of the course (P < 0.05). Nasal epithelial micronuclei increased 22%, from 0.41 +/- 0.52/1000 cells to 0.50 +/- 0.67/1000 cells (P = 0.26). In blood cells, the frequency of micronucleated lymphocytes increased 28%, from 4.95 +/- 1.72/1000 cells to 6.36 +/- 2.03/1000 cells (P < 0.05), while sister chromatid exchanges decreased 7.5% (P < 0.05). A dose-response relationship was observed between cumulative exposure to formaldehyde and increases in buccal micronuclei in the 22 male subjects but not in the 7 female subjects. We conclude that low-level exposure to formaldehyde is associated with cytogenetic changes in epithelial cells of the mouth and in blood lymphocytes. These cytogenetic effects may be useful as markers of biologically effective dose.

Adult↗