Toxic Formaldehyde Research Articles

Degradation of plastic polyoxymethylene brackets and the subsequent release of toxic formaldehyde

Heat, acids, alkalis, oxygen, abrasion, enzymes, and radiation are all viable mechanisms for the chemical breakdown of polyoxymethylene (POM), a plastic material used in some esthetic orthodontic brackets. The aim of this study was to establish the thermal characteristics of POM brackets and the chemical by-products in the as-received bracket, during thermal analyses of the bracket, and after abrasion of the slot and base of the bracket. Plastic brackets and control rods made of POM were evaluated ex vivo by thermal and chemical analyses. POM brackets produce toxic formaldehyde gas whether heated or mechanically abraded. Patients who wear these brackets are being exposed to, at the very least, a potential irritant. Thermal analyses showed that the melting temperature of a POM bracket was approximately 178 degrees C, comparable to that for a commercial rod of POM. Both POM products started to degrade at approximately 250 degrees C, and, by 420 degrees C, both products had completely decomposed into their fundamental molecular structure, formaldehyde. A colorimetric assay with a fuchsin-aldehyde reagent (Schiff's reagent) showed that aldehydes were present in the effluent from thermal heating, from mechanical abrasion of the bracket's slot or base, and even from the as-received bracket. The only difference between the 6 to 8-week assays and the 12 to 14-week assays was the intensification of the color, which occurred because of an increase in aldehyde formation and the concomitant increase in double-bond formation. These observations are consistent with a report in the tribologic (ie, friction and wear) literature, which documented the degradation of POM when it rubs against steel. Because formaldehyde's inherent uses as a disinfectant base and embalming fluid preclude its beneficial presence in the human body, further use of POM for orthodontic brackets, crowns for children, and other prosthetic appliances is contraindicated because even radiography will promote its degradation.

Formaldehyde toxicity is altered by calcium and organic matter

Summary Formaldehyde is one of the shortlist substances permitted for use in aquaculture. Toxicity of formaldehyde and therapeutical success is influenced by water parameters. However, there is a lack of information on the interaction of organic carbon and calcium (Ca2+) on the toxicity of formaldehyde. Laboratory experiments were conducted to test the combined effects of Ca2+ and natural organic matter (NOM) on formaldehyde toxicity. Zebrafish embryos were exposed to five concentrations of formaldehyde for 144 h. Juveniles exposed to high concentrations died; the embryos exposed to low concentrations were unable to hatch, particularly visible in the groups without NOM. The toxicity-reducing effects of NOM are not attributable to the presence of Ca2+.

Vitamin E against oxidative damage caused by formaldehyde in frontal cortex and hippocampus: Biochemical and histological studies

Formaldehyde (FA) can cause severe central nervous system impairment. But, there are only a few studies about biochemical and histopathological changes of frontal cortex and hippocampal tissue caused by FA toxicity. The aim of our study was to investigate these changes occurring after chronic formaldehyde toxicity in frontal cortex and hippocampal tissues, and protective effect of Vitamin E (vit E) against oxidative damage. Eighteen rats were divided into three groups: (1) control, (2) treated with FA (FAt), and (3) treated with FA and vit E (FAt + vit E) groups. After the treatment, the animals were sacrificed and frontal cortex and hippocampal tissues were removed for biochemical and histopathological investigation. FA significantly increased tissue malondialdehyde (MDA) and protein carbonyl (PC) levels and also decreased superoxide dismutase (SOD) and catalase (CAT) enzyme activities in frontal cortex and hippocampal tissue compared to control. Vit E treatment decreased MDA and PC levels and prevented inhibition of SOD and CAT enzymes in the tissues. In the FAt group, the neurons of both tissues became extensively dark and degenerated with picnotic nuclei. The morphology of neurons in FAt + vit E group was protected well, but not as neurons of the control group. The number of neurons in frontal cortex and hippocampal tissue of FAt group was significantly less than both control and FAt + vit E groups. It was concluded that vit E treatment might be beneficial in preventing FA-induced oxidative frontal cortex and hippocampal tissue damage, therefore, shows potential for clinical use.

Assimilation, dissimilation, and detoxification of formaldehyde, a central metabolic intermediate of methylotrophic metabolism

Methanol is a valuable raw material used in the manufacture of useful chemicals as well as a potential source of energy to replace coal and petroleum. Biotechnological interest in the microbial utilization of methanol has increased because it is an ideal carbon source and can be produced from renewable biomass. Formaldehyde, a cytotoxic compound, is a central metabolic intermediate in methanol metabolism. Therefore, microorganisms utilizing methanol have adopted several metabolic strategies to cope with the toxicity of formaldehyde. Formaldehyde is initially detoxified through trapping by some cofactors, such as glutathione, mycothiol, tetrahydrofolate, and tetrahydromethanopterin, before being oxidized to CO2. Alternatively, free formaldehyde can be trapped by sugar phosphates as the first reaction in the C1 assimilation pathways: the xylulose monophosphate pathway for yeasts and the ribulose monophosphate (RuMP) pathway for bacteria. In yeasts, although formaldehyde generation and consumption takes place in the peroxisome, the cytosolic formaldehyde oxidation pathway also plays a role in formaldehyde detoxification as well as energy formation. The key enzymes of the RuMP pathway are found in a variety of microorganisms including bacteria and archaea. Regulation of the genes encoding these enzymes and their catalytic mechanisms depend on the physiological traits of these organisms during evolution.

Different cytoprotective effect of antioxidants and change in the iron regulatory system in rodent cells exposed to paraquat or formaldehyde

To study the mechanism of toxicity of paraquat and formaldehyde, the response of oxidant-exposed cultured NIH3T3 cells to antioxidants or an iron chelator was investigated. Paraquat-induced cell death was reduced by treatment with 10 microM pyrrolidine dithiocarbamate (PDTC) and 10 microM desferrioxamine (DFO), but not with N-acetyl-L-cysteine (NAC). Cells were protected from formaldehyde-induced cytotoxicity by 1 mM NAC, but not by PDTC or DFO. Moreover, paraquat modulated the cellular iron regulatory system. Paraquat induced a time-dependent increase in the binding of iron regulatory protein 1 (IRP1) to iron-responsive element (IRE), and the enhanced IRP1 activity continued over 24 h. On the other hand, no induction of increased IRP1 binding to IRE was observed in rodent cells exposed to formaldehyde. Previously, we observed stimulation of EpRE-mediated ferritin mRNA expression in the cells exposed to hydrogen peroxide. However, paraquat did not induce any transcriptional activation of ferritin genes. These results suggest that intracellular iron may be involved in paraquat-mediated cytotoxicity and the influence of paraquat on iron metabolism differs from that of hydrogen peroxide.

Fomepizole as a therapeutic strategy in paediatric methanol poisoning. A case report and review of the literature

Methanol poisoning is not frequently observed in children; however, without treatment, serious intoxication can be complicated by visual impairment, coma, metabolic acidosis, respiratory and circulatory insufficiency and death. Treatment in a paediatric intensive care is therefore compulsory. Methanol is metabolised in the liver by alcohol dehydrogenase to the toxic metabolites formaldehyde and formic acid. Classically, ethanol is given as a competitive inhibitor in order to avoid the formation of these compounds. We report on the use of fomepizole (4-methylpyrazole),a new and potent inhibitor of alcohol dehydrogenase, in a 3-year-old boy after the intake of a toxic amount of methanol. The course was uneventful and the use of fomepizole was not accompanied by any side-effects. An overview is given of all cases of paediatric poisoning in which fomepizole was used. Fomepizole seems to be a safe and valid alternative to ethanol in cases of paediatric methanol poisoning.

Formaldehyde degradation in an anaerobic packed-bed bioreactor

The development of appropriate technologies for the treatment of formaldehyde discharged into the environment is important to minimize its impact. Aerobic systems have been employed, although alternative anaerobic treatments have also been widely studied, mainly due to their low energy consumption and sludge production. However, toxic substances can lead to disturbances in anaerobic reactors. Some research has already been developed on formaldehyde anaerobic biological treatment, but no consensus has yet been reached about its behavior nor has the most efficient system been identified. Aiming at finding supporting evidence for this issue, therefore, this study investigated the degradation and toxicity of formaldehyde in a Horizontal-Flow Anaerobic Immobilized Sludge Reactor. Formaldehyde concentrations of 26.2–1158.6 mg HCHO/L were applied in the reactor, resulting in formaldehyde and chemical oxygen demand removal efficiencies of 99.7% and 92%, respectively. Volatile fatty acids with up to five carbons, found during the degradation of formaldehyde, are believed to indicate that the degradation followed routes unlike those suggested in the literature, which reports the formation of intermediates such as methanol and formic acid. The Monod kinetic model adhered to the experimental data well, with apparent kinetic parameters estimated as r max app=2.79×10 −3 mg HCHO/mg SSV h and K s app=242.8 mg HCHO/L.

Involvement of SSAO-mediated deamination in adipose glucose transport and weight gain in obese diabetic KKAy mice.

Semicarbazide-sensitive amine oxidase (SSAO) is located on outer surfaces of adipocytes and endothelial and vascular smooth muscle cells. This enzyme catalyzes deamination of methylamine and aminoacetone, leading to production of toxic formaldehyde and methylglyoxal, respectively, as well as hydrogen peroxide and ammonium. Several lines of evidence suggest that increased SSAO activity is related to chronic inflammation and vascular disorders related to diabetic complications. We found that a highly potent and selective SSAO inhibitor, (E)-2-(4-fluorophenethyl)-3-fluoroallylamine (FPFA), was capable of reducing numbers of atherosclerotic lesions as well as weight gain in obese KKAy mice fed an atherogenic diet. SSAO inhibitors cause a moderate and long-lasting hyperglycemia. Such an increase in serum glucose is a result of reduction of glucose uptake by adipocytes. SSAO-mediated deamination of endogenous methylamine substrates induces adipocyte glucose uptake and lipogenesis. Highly selective SSAO inhibitors can effectively block induced glucose uptake. The results suggest that increased SSAO-mediated deamination may be concomitantly related to obesity and vascular disorders associated with type 2 diabetes.

Channelling and formation of 'active' formaldehyde in dimethylglycine oxidase.

Here we report crystal structures of dimethylglycine oxidase (DMGO) from the bacterium Arthrobacter globiformis, a bifunctional enzyme that catalyzes the oxidation of N,N-dimethyl glycine and the formation of 5,10-methylene tetrahydrofolate. The N-terminal region binds FAD covalently and oxidizes dimethylglycine to a labile iminium intermediate. The C-terminal region binds tetrahydrofolate, comprises three domains arranged in a ring-like structure and is related to the T-protein of the glycine cleavage system. The complex with folinic acid indicates that this enzyme selectively activates the N10 amino group for initial attack on the substrate. Dead-end reactions with oxidized folate are avoided by the strict stereochemical constraints imposed by the folate-binding funnel. The active sites in DMGO are approximately 40 A apart, connected by a large irregular internal cavity. The tetrahydrofolate-binding funnel serves as a transient entry-exit port, and access to the internal cavity is controlled kinetically by tetrahydrofolate binding. The internal cavity enables sequestration of the reactive iminium intermediate prior to reaction with tetrahydrofolate and avoids formation of toxic formaldehyde. This mode of channelling in DMGO is distinct from other channelling mechanisms.

Enzyme-catalyzed polymerization to functional polymers

In this article, described are our recent advances in enzymatic polymerization, defined as chemical polymer syntheses in vitro (in test tubes) via non-biosynthetic pathways catalyzed by an isolated enzyme. The major target macromolecules formed via the enzymatic polymerizations in this article are polyesters and polyphenols. For synthesis of polyesters, hydrolases are used as catalyst; hydrolases, enzymes catalyzing a bond-cleavage reaction by water, induce the reverse reaction of hydrolysis, leading to polymer production by a bond-forming reaction. Specific enzyme catalysis provides a novel synthetic route for useful polyesters, many of which are difficult to be synthesized by conventional methodologies. Peroxidase and laccase act as catalyst for oxidative polymerization of various phenol derivatives to produce a new class of phenolic polymers without use of toxic formaldehyde under mild reaction conditions. Artificial urushi has been developed by laccase-catalyzed curing of new urushiol analogues.

Formaldehyde cytotoxicity in three human cell types assessed in three different assays

International standards for preclinical screening of the cytotoxicity of dental materials so far recommend the use of established cell lines. The aim of this study was to assess the relative susceptibility of human dental pulp fibroblasts (HPF), human buccal epithelial cells (HBE) and HeLa cervix cancer cells exposed to identical cytotoxic challenges. Formaldehyde, which may be released from dental materials such as dental composites, glassionomer cements, and endodontic sealers, was used as test chemical. Cytotoxicity data including dose–response relations and TC 50 values were assessed in three different assays: BrdU incorporation, neutral red uptake and MTT assays. HBE and HPF demonstrated statistically significant lower TC 50 values in both the neutral red and the BrdU assay in comparison to HeLa cells. In the MTT assay no statistically significant differences were observed between the cell types. In the two target-tissue cell types (HPF and HBE) the Neutral Red assay revealed lower TC 50 values in comparison to the BrdU assay. In HeLa cells no statistically significant differences were observed between the assays. In conclusion, the present study confirms that cytotoxicity data obtained by cell culture studies are influenced by both cell culture model and choice of assay. Under identical experimental conditions, human target tissue cells appeared to be more sensitive to formaldehyde toxicity than human HeLa cancer cells.

Self-Association of m-Cresol in Aqueous Organic Solvents: Relation to Enzymatic Polymerization Reaction

Oxidative polymerization of phenols using horseradish peroxidase (HRP) in aqueous organic solvents (1,4-dioxane-water and methanol-water), which is an attractive pathway for synthesis of a new class of polyphenols withoutuse of toxic formaldehyde, was remarkably influenced by the water content. The present study is focused on the self-association of m-cresol in such a solvent to understand the effect of solvent on its polymerization. The change of the molecular weight of the polymer with varying the water content was found to be related to the molecular clustering in the solution, observed through the difference UV absorption spectroscopy and the mass spectrometry for clusters. In mixed solvents of 1,4-dioxane and aqueous 0. 1 M phosphate buffer (pH 7), number-average molecular weight had a maximum at the aqueous buffer content of 50 vol %, whereas in methanol-aqueous buffer mixtures, the molecular weight had a small maximum at 20 vol % aqueous buffer content and gradually decreased with increase of the aqueous buffer content. As for the m-cresol clustering, the effect of the water content was also in significant contrast between the above aqueous organic solvent systems. The m-cresol clusters were not formed in 1,4-dioxane, but its clustering was remarkably promoted by an increase of the water content. On the other hand, the m-cresol clusters were formed favorably in methanol and were readily hydrated with increase of the water content in the solution. These effects of the water content on the clustering are related to the enzymatic polymerization. The polymer would be obtained in high yields when the m-cresol clusters were formed in the solution; however, the hydration of the m-cresol cluster would restrain the polymerization.

Derivation of ambient water quality criteria for formaldehyde

This paper describes the derivation of aquatic life water quality criteria for formaldehyde, developed in accordance with United States Environmental Protection Agency's (USEPA's) Guidelines for Deriving Numerical National Water Quality Criteria for the Protection of Aquatic Organisms and Their Uses. The initial step in deriving water quality criteria was to conduct an extensive literature search to assemble available acute and chronic toxicity data for formaldehyde. The literature search identified a large amount of information on acute toxicity of formaldehyde to fish and aquatic invertebrates. These acute data were evaluated with respect to data quality, and poor quality or uncertain data were excluded from the data base. The resulting data base met the USEPA requirements for criteria derivation by having data for at least one species in at least eight different taxonomic families. One shortcoming of the literature-derived data base, however, was that few studies involved analytical confirmation of nominal formaldehyde concentrations and reported toxicity endpoints. Also, there were relatively few data on chronic toxicity. The acute toxicity data set consisted of data for 12 species of fish, 3 species of amphibians, and 11 species of invertebrates. These data were sufficient, according to USEPA guidelines, to calculate a final acute value (FAV) of 9.15 mg/l, and an acute aquatic life water quality criterion (one-half the FAV) of 4.58 mg/l. A final acute–chronic ratio (ACR) was calculated using available chronic toxicity data and USEPA-recommended conservative default assumptions to account for missing data. Using the FAV and the final ACR (5.69), the final chronic aquatic life water quality criterion was determined to be 1.61 mg/l.

Embryo toxicity and teratogenicity of formaldehyde.

C-14 formaldehyde crosses the placenta and enters fetal tissues. The incorporated radioactivity is higher in fetal organs (i.e., brain and liver) than in maternal tissues. The incorporation mechanism has not been studied fully, but formaldehyde enters the single-carbon cycle and is incorporated as a methyl group into nucleic acids and proteins. Also, formaldehyde reacts chemically with organic compounds (e.g., deoxyribonucleic acid, nucleosides, nucleotides, proteins, amino acids) by addition and condensation reactions, thus forming adducts and deoxyribonucleic acid-protein crosslinks. The following questions must be addressed: What adducts (e.g., N-methyl amino acids) are formed in the blood following formaldehyde inhalation? What role do N-methyl-amino adducts play in alkylation of nuclear and mitochondrial deoxyribonucleic acid, as well as mitochondrial peroxidation? The fact that the free formaldehyde pool in blood is not affected following exposure to the chemical does not mean that formaldehyde is not involved in altering cell and deoxyribonucleic acid characteristics beyond the nasal cavity. The teratogenic effect of formaldehyde in the English literature has been sought, beginning on the 6th day of pregnancy (i.e., rodents) (Saillenfait AM, et al. Food Chem Toxicol 1989, pp 545-48; Martin WJ. Reprod Toxicol 1990, pp 237-39; Ulsamer AG, et al. Hazard Assessment of Chemicals; Academic Press, 1984, pp 337-400; and U.S. Department of Health and Human Services. Toxicological Profile of Formaldehyde; ATSDR, 1999 [references 1-4, respectively, herein]). The exposure regimen is critical and may account for the differences in outcomes. Pregnant rats were exposed (a) prior to mating, (b) during mating, (c) or during the entire gestation period. These regimens (a) increased embryo mortality; (b) increased fetal anomalies (i.e., cryptochordism and aberrant ossification centers); (c) decreased concentrations of ascorbic acid; and (d) caused abnormalities in enzymes of mitochondria, lysosomes, and the endoplasmic reticulum. The alterations in enzymatic activity persisted 4 mo following birth. In addition, formaldehyde caused metabolic acidosis, which was augmented by iron deficiency. Furthermore, newborns exposed to formaldehyde in utero had abnormal performances in open-field tests. Disparities in teratogenic effects of toxic chemicals are not unusual. For example, chlorpyrifos has not produced teratogenic effects in rats when mothers are exposed on days 6-15 (Katakura Y, et al. Br J Ind Med 1993, pp 176-82 [reference 5 herein]) of gestation (Breslin WJ, et al. Fund Appl Toxicol 1996, pp 119-30; and Hanley TR, et al. Toxicol Sci 2000, pp 100-08 [references 6 and 7, respectively, herein]). However, either changing the endpoints for measurement or exposing neonates during periods of neurogenesis (days 1-14 following birth) and during subsequent developmental periods produced adverse effects. These effects included neuroapoptosis, decreased deoxyribonucleic acid and ribonucleic acid synthesis, abnormalities in adenylyl cyclase cascade, and neurobehavioral effects (Johnson DE, et al. Brain Res Bull 1998, pp 143-47; Lassiter TL, et al. Toxicol Sci 1999, pp 92-100; Chakraborti TK, et al. Pharmacol Biochem Behav 1993, pp 219-24; Whitney KD, et al. Toxicol Appl Pharm 1995, pp 53-62; Chanda SM, et al. Pharmacol Biochem Behav 1996, pp 771-76; Dam K, et al. Devel Brain Res 1998, pp 39-45; Campbell CG, et al. Brain Res Bull 1997, pp 179-89; and Xong X, et al. Toxicol Appl Pharm 1997, pp 158-74 [references 8-15, respectively, herein]). Furthermore, the terata caused by thalidomide is a graphic human example in which the animal model and timing of exposure were key factors (Parman T, et al. Natl Med 1999, pp 582-85; and Brenner CA, et al. Mol Human Repro 1998, pp 887-92 [references 16 and 17, respectively, herein]). Thus, it appears that more sensitive endpoints (e.g., enzyme activity, generation of reactive oxygen species, timing of exposure) for the measurement of toxic effects of environmental agents on embryos, fetuses, and neonates are more coherent than are gross terata observations. The perinatal period from the end of organogenesis to the end of the neonatal period in humans approximates the 28th day of gestation to 4 wk postpartum. Therefore, researchers must investigate similar stages of development (e.g., neurogenesis occurs in the 3rd trimester in humans and neonatal days occur during days 1-14 in rats and mice, whereas guinea pigs behave more like humans). Finally, screening for teratogenic events should also include exposure of females before mating or shortly following mating. Such a regimen is fruitful inasmuch as environmental agents cause adverse effec

The formaldehyde metabolic detoxification enzyme systems and molecular cytotoxic mechanism in isolated rat hepatocytes

The toxicity and carcinogenicity of formaldehyde (HCHO) has been attributed to its ability to form adducts with DNA and proteins. A marked decrease in mitochondrial membrane potential and inhibition of mitochondrial respiration that was accompanied by reactive oxygen species formation occurred when isolated rat hepatocytes were incubated with low concentrations of HCHO in a dose-dependent manner. Hepatocyte GSH was also depleted by HCHO in a dose-dependent manner. At higher HCHO concentrations, lipid peroxidation ensued followed by cell death. Cytotoxicity studies were conducted in which isolated hepatocytes exposed to HCHO were treated with inhibitors of HCHO metabolising enzymes. There was a marked increase in HCHO cytotoxicity when either alcohol dehydrogenase or aldehyde dehydrogenase was inhibited. Inhibition of GSH-dependent HCHO dehydrogenase activity by prior depletion of GSH markedly increased hepatocyte susceptibility to HCHO. In each case, cytotoxicity was dose-dependent and corresponded with a decrease in hepatocyte HCHO metabolism and increased lipid peroxidation. Antioxidants and iron chelators protected against HCHO cytotoxicity. Cytotoxicity was also prevented, when cyclosporine or carnitine was added to prevent the opening of the mitochondrial permeability transition pore which further suggests that HCHO targets the mitochondria. Thus, HCHO-metabolising gene polymorphisms would be expected to have toxicological consequences on an individual's susceptibility to HCHO toxicity and carcinogenesis.

Formaldehyde toxicity in anaerobic systems

Formaldehyde toxicity during acetoclastic methanogenesis exerted a dual character: It was in part reversible and in part irreversible. Toxicity was only reversible at very low formaldehyde concentration. The degree of irreversible toxicity was proportionally related to the initial sludge concentration, but was independent of the initial specific acetoclastic activity. Formaldehyde conversion seemed to be dependent on the concentration of the various trophic groups present in the anaerobic granular sludge used. Hence biomass diversity may play an important role in the outcome of toxicity studies and may furthermore contribute to the explanation of the controversies with respect to toxicity values reported in literature.

Histological distribution of class III alcohol dehydrogenase in human brain

The distributions of class III alcohol dehydrogenase (ADH), a glutathione-dependent formaldehyde dehydrogenase, and class I ADH in the human brain were examined immunohistochemically. The most intense immunostaining of class III ADH was observed in the dendrites and cytoplasm of cerebellar Purkinje cells. Scattered cerebral cortical neurons in layers IV and V, and some hippocampal pyramidal neurons were also immunopositive. The neuronal distribution of class III ADH resembled that of the vulnerable neurons in patients with hypoxic encephalopathy, which in view of the intense staining in the Purkinje cells, raises the possibility that this enzyme contributes to the hypoxia and cerebellar degeneration suffered by chronic alcoholics. Perivascular and subependymal astrocytes, which contribute to the maintenance of the cerebral cellular milieu and isolate the brain from the systemic circulation and cerebrospinal fluid, were also class III ADH positive. As the substrates of this enzyme include intrinsic toxic formaldehyde, inflammatory intermediate of 20-hydroxy-leukoteiene B4, and possibly ethanol, the distribution of class III ADH immunostaining indicates this enzyme contributes to the defence of the brain against degenerative processes. The finding that, unlike ependymal cells, subependymal astrocytes were class III ADH positive, suggests this enzyme may be useful for differentiating astrocytes and ependymal cells.

Continuous anaerobic treatment of wastewaters containing formaldehyde and urea

The toxicity of formaldehyde (FA) in batch assays, using volatile fatty acids (VFA) as co-substrate, and the continuous anaerobic treatment of wastewaters containing FA in upflow anaerobic sludge blanket (UASB) reactors was investigated. In batch studies, FA exerted a 50% methanogenic toxicity on VFA at concentrations of around 100 mg/l, 2.5 times lower than values reported with sucrose. Although at FA concentrations higher than 200 mg/l methanogenesis was completely inhibited, a partial recovery of the bacterial activity was observed after 250 h when the FA had been removed from the medium. The continuous anaerobic degradation of FA at concentrations up to 2 g/l, using 1.6 g/l of glucose as co-substrate, was studied in a UASB reactor. A stable and efficient operation was observed at organic loading rates (OLR) of 6.0 g COD/l·d and with a COD/FA ratio as low as 1.4. A synthetic substrate with the same characteristics as the effluents produced during fibreboard adhesives manufacturing (based on urea-FA), i.e. 0.95 g FA/l and 0.35 g urea/l, was treated in a UASB reactor. The applied OLR and nitrogen loading rate (NLR) were 3.45 g COD/l·d and 0.58 g N/l·d, respectively. COD removal efficiencies were maintained at 90–95%, FA and urea being completely degraded.

Toxicity Effects of Formaldehyde on Methanol Degrading Sludge and Its Anaerobic Conversion in Biobed® Expanded Granular Sludge Bed (EGSB) Reactors

Methanogenesis from formaldehyde mainly occurred via intermediates as confirmed by the increased concentrations of methanol and H2 in the liquid and gas phases respectively during formaldehyde conversion. While formaldehyde was readily transformed, the methane production rate was immediately and strongly inhibited. Formaldehyde toxicity was in part reversible since the methane production rate recovered after formaldehyde depletion. This recovery can not be explained by biomass growth. The toxicity of formaldehyde was also in part irreversible, since the degree of recovery was not complete. This loss in the methane production rate likely can be attributed to biomass decay as suggested by its linear relation with the amount of formaldehyde dosed, as well as by scanning electron microscopy observations. The addition of the same amount of formaldehyde either in a slug or in a continuous mode caused the same loss in the methane production rate. Thus for the treatment of formaldehyde-containing streams, a balance between loss in the rate (formaldehyde-related decay) and bacterial growth should be attained. By combining good biomass retention and internal dilution of the wastewater, industrial streams containing formaldehyde can still be treated anaerobically.

How overproduction of foreign proteins affects physiology of the recombinant strains ofHansenula polymorpha

Changes in the activity of key enzymes of the methanol utilization pathway of the recombinant strains of methylotrophic yeastHansenula polymorpha R22-2B and LAC-56 were studied at different rates of chemostat growth on methanol containing mineral media. It was shown that the strain R22-2B, initially having a 10-fold increased activity of dihydroxyacetone kinase (DHAK, a key enzyme of formaldehyde assimilation) acquired increased activity of formaldehyde dehydrogenase (FADH, a key enzyme of formaldehyde dissimilation) which resulted in the enhanced oxidation of formaldehyde to CO2. Strain LAC-56, overproducingEscherichia coli β-galactosidase, acquired the decreased intracellular concentration of ATP which resulted in the decrease of the efficiency of formaldehyde assimilation catalyzed by DHAK and resulted in accumulation of toxic formaldehyde. As a consequence some biochemical responses occurred in cells that were directed to a diminishing of the toxic effect of accumulated formaldehyde, namely, the decreasing of methanol oxidase activity (to reduce the rate of formaldehyde synthesis), and the increasing of FADH activity (to increase the rate of formaldehyde oxidation).