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B Holmbom

Publications and source records attributed to B Holmbom.

15 recordsLinked to original sources

Fate of wood extractives in wastewater treatment plants at kraft pulp mills and mechanical pulp mills.

Extensive environmental effects of the forest industry led to implementation of activated sludge treatment of effluents in the 1980s. Although the existence of chlorinated compounds in the effluents has decreased, a discussion about the possible environmental effects of elemental-chlorine-free (ECF) and total-chlorine-free (TCF) bleached pulp mill effluents has arisen, and chronic effects on aquatic organisms have still been found. Recently, studies have mainly focussed on wood extractives and their role in the effects of effluents. Resin acids and unsaturated fatty acids are found to be toxic, and plant sterols are reported to have adverse endocrine effects on water organisms already at low concentrations. In this study, Finnish wastewater treatment plants of an ECF kraft pulp mill, a paper mill, and an integrated TCF kraft pulp and paper mill were sampled in order to ascertain how wastewater treatment plants, and especially activated sludge treatments, remove wood extractives. Concentrations of extractives in discharged wastewaters varied between 0.4 and 11 g/t kraft or mechanical pulp, and the concentrations decreased over 95% during the treatment processes. Of the wood extractives, 1.1-64% were adsorbed to biosludge and 35-99% were degraded or transformed to other forms during the activated sludge treatment. A major part of these compounds were discharged in particles (74-99%). The removal of extractives was efficient even in the effluent treatment plant, which was highly loaded during the sampling period.

Environmental Monitoring↗

Hydrolysis of steryl esters by a lipase (Lip 3) from Candida rugosa.

A well-known lipase, Lip 3 of Candida rugosa, was purified to homogeneity from a commercial lipase preparation, using hydrophobic interaction and anion exchange chromatography. Lip 3, which has been reported to act on cholesteryl esters, was also found to be active on plant-derived steryl esters. Lip 3 had optimal activity at pH 5-7 and below 55 degrees C. It was able to hydrolyse steryl esters totally in a clear micellar aqueous solution. However, the action on a dispersed colloidal steryl ester solution was limited and only about half of the steryl esters were degraded. The degree of hydrolysis was not improved by addition of fresh enzyme. The composition of released fatty acids and sterols was, however, almost identical to that obtained by alkaline hydrolysis, showing that all the different steryl esters were hydrolysed equally and that none of the individual components were responsible for incomplete hydrolysis. Thus, it appeared that the physical state of the colloidal steryl ester dispersion limited the action of Lip 3. Wood resins contain both triglycerides and steryl esters among the hydrophobic components, which create problems in papermaking. The simultaneous enzymatic hydrolysis of triglycerides and steryl ester is therefore of considerable interest and Lip 3 is the first enzyme reported to act on both triglycerides and steryl esters.

Candida↗

Reactivity of Trametes laccases with fatty and resin acids.

Lipophilic extractives commonly referred to as wood pitch or wood resin can have a negative impact on paper machine runnability and product quality. The lipophilic extractives are composed mainly of fatty acids, resin acids, sterols, steryl esters and triglycerides. In this work, the suitability of laccases for the modification of fatty and resin acids was studied, using two model fractions. In the treatments, resin and fatty acid dispersions were treated with two different laccases, i.e. laccases from Trametes hirsuta and T. villosa. Different chromatographic methods were used to elucidate the effects of laccase treatments on the chemistry of the fatty and resin acids. Both laccases were able to modify the fatty and resin acids to some extent. In the case of fatty acids, a decrease in the amount of linoleic, oleic and pinolenic acids was observed, whereas the modification of resin acids resulted in a reduced amount of conjugated resin acids.

Abietanes↗

Expression of the vitellogenin gene in the liver of juvenile whitefish (Coregonus lavaretus L. s.l.) exposed to effluents from pulp and paper mills.

Juvenile whitefish (Coregonus lavaretus L. s.l.) were exposed by caging in the field to diluted effluents from three operating pulp, paper, and paperboard mills in Southern Lake Saimaa, Finland. The expression of the vitellogenin gene, used as a biomarker of estrogenic contamination of effluents, was measured using a Northern blotting method. Increased mRNA levels, the most specific and reliable evidence for estrogen receptor-mediated actions in vivo, were found in fish caged in the vicinity of one of three mills studied. This mill was found to discharge wood-derived compounds, such as sterols and resin acids, into Lake Saimaa in amounts considerably exceeding those from the other two mills. The increased vitellogenin gene expression suggests that the effluent is a source of estrogenic contaminants.

Animals↗

Wood-derived estrogens: studies in vitro with breast cancer cell lines and in vivo in trout.

The wood-derived compound, beta-sitosterol (purity > 90%), was shown to be estrogenic in fish. It induced the expression of the vitellogenin gene in the liver of juvenile and methyltestosterone-treated rainbow trout. Structural similarities to beta-sitosterol notwithstanding, cholesterol, citrostadienol, beta-sitostanol, and 5-androstene-3 beta,17 beta-diol, an estrogenic member of the androstenic steroid group, were inactive. An abietic acid mixture (37% abietic acid, 6% dehydroabietic acid, and a remainder of unknown compounds) showed slight hormonal activity in feed, but it was completely inactive when given intraperitoneally in implants. The estrogenic component of the abietic acid preparation was not identified. In addition, to beta-sitosterol and abietic acid, several other wood-derived compounds including betulin, isorhapontigenin, isorhapontin, and pinosylvin were estrogenic in breast cancer cells (MCF-7 or T-47D). However, betulin and pinosylvin, available in sufficient amounts for in vivo testing, did not induce the expression of the vitellogenin gene. Differences in the primary sequences of human and fish estrogen receptors (hormone as well as DNA-binding regions) or uptake and metabolism of the compounds may explain the discrepancy between the two estrogen bioassays. Wood-derived compounds such as beta-sitosterol, present in pulp and paper mill effluents, may account for the weak estrogenicity of debarking effluent seen at the vitellogenin expression bioassay.

Animals↗

Comparison of triphenyltetrazolium chloride (TTC) staining versus detection of fibronectin in experimental myocardial infarction.

Staining with triphenyltetrazolium chloride (TTC), although controversial, has frequently been used for the delineation of myocardial infarction. This study was performed further to explore the reliability of the TTC method. In 24-h experiments pigs were subjected to closed-chest occlusion of the left anterior descending coronary artery for 30, 60 or 90 min followed by reperfusion with or without superoxide dismutase (SOD) as an adjunct. One TTC-stained slice from each heart was stabilized by microwave irradiation, gelatin-embedded, frozen in hexane chilled with dry ice and cryosectioned. Serial sections were stained with antibodies against fibronectin in order to identify irreversibly injured myocytes and with van Gieson histologically to confirm the necrotic tissue. A close correspondence of the infarct size was found between TTC stained slices and anti-fibronectin stained sections. The infarct size in the van Gieson stained sections also showed good correspondence but the area of infarction tended to be larger. In the experimental group subjected to 30 min ischaemia and with SOD as an adjunct, the estimated infarcted area in the TTC stained slices was significantly smaller than the area estimated from the anti-fibronectin stained sections. In sections viewed in the light microscope an inverse pattern of TTC and anti-fibronectin staining was observed. It was confirmed at the light microscopic level that myocytes containing an abundance of TTC deposits lacked fibronectin whereas myocytes stained with antifibronectin in general lacked TTC staining except for a zone approximately 0.5 mm wide which was located at the intersection between damaged and surviving myocytes where small TTC deposits were present. The width of the stained zone did not differ among the experimental groups. Thus, differences in estimated infarct size by the three methods used reflect problems in correctly delineating the border between living and dead myocardium rather than an interference by SOD on TTC staining.

Animals↗

Enzyme and immunohistochemical assessment of myocardial damage after ischaemia and reperfusion in a closed-chest pig model.

The usefulness of different enzyme and immunohistochemical stains to distinguish reversible and irreversible myocardial cell injury after experimental coronary artery occlusion of varying duration and reperfusion with or without superoxide dismutase as adjunct was investigated. Biopsies or parts of the infarcted and non-infarcted area were rapidly frozen and sectioned in series for enzyme and immunohistochemical evaluation. Sections were stained for the demonstration of phosphorylase, myofibrillar ATPase and mitochondrial oxidative enzymes and also with periodic acid-Schiff, alizarin red S and routine histological stains. Other sections in series were stained with antibodies against fibronectin and the intermediate filament proteins desmin and vimentin. In 49 biopsies a blind quantitative estimation of the area stained for fibronectin, phosphorylase and alizarin red S was performed and evaluated statistically. Phosphorylase, periodic acid-Schiff, fibronectin and alizarin red S allowed delineation of affected myocardium after 30 min of ischaemia followed by reperfusion whereas with the other stains, affected myocardium was readily detectable only after 60 or 90 min of ischaemia followed by reperfusion as well as after 24 h of ischaemia without reperfusion. The immunostaining for fibronectin was very distinct and inversely related to the phosphorylase activity. We show that fibronectin is an excellent marker for damaged cells and that these positively stained myocytes are necrotic as confirmed ultrastructurally. Using alizarin red S as a marker of calcium accumulation in myocytes, a marked discrepancy was observed between the area of fibronectin-containing myocytes and that of myocytes stained by alizarin red S. Calcium accumulation in mitochondria is thus not a prerequisite for myocyte necrosis but does occur only in some of the irreversibly damaged cells. Of special interest is the finding that there was a significant reduction of intracellular calcium in pigs where superoxide dismutase had been used as an adjunct at reperfusion, thus supporting the theory that free radicals do play a role during reperfusion of ischaemic myocardium.

Adenosine Triphosphatases↗

A method for enzyme- and immunohistochemical staining of large frozen specimens.

A method for large specimen cryosectioning is described. Specimens of pig heart ventricles were lightly fixed by microwave irradiation, embedded in 10% gelatin, frozen in hexan chilled with dry ice, and sectioned using an LKB 2250 PMW cryomicrotome. The sections were collected on transparent film and transferred onto glass slides. Standard histological, enzyme- and immuno-histochemical staining techniques were used. The present method allowed cryosectioning with sections of good quality which could be used for enzyme- and immuno-staining. For studies of experimental myocardial infarction, staining for phosphorylase and Periodic Acid Schiff outlined the ischemic area and antibodies against plasma fibronectin and fibrinogen delineated the infarcted myocardium.

Animals↗

Mutagenic compounds in chlorinated pulp bleaching waters and drinking waters.

A review is given of the isolation and identification of the extremely strong bacterial mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone, coded MX, in wood pulp chlorination waters in 1980. This compound is one of the most potent known mutagens, as determined by the Ames test. In 1986 this compound was found also in chlorinated drinking water. MX accounts for a substantial portion of the mutagenicity of chlorinated drinking waters. Several other compounds related to MX have recently been identified in chlorinated waters, such as the geometric isomer (E-MX), the oxidized and reduced forms of MX and the oxidized form of E-MX. Brominated analogues of MX may be formed by chlorination in the presence of bromide ions. Unanswered questions pertaining to the chemistry and genotoxicity of MX and related chlorinated mutagens are examined.

Furans↗

Myomesin and M protein: differential expression in embryonic fibers during pectoral muscle development.

By applying immunocytochemistry using monoclonal antibodies, we found that the myofibrillar M band of both presumptive type-I and -II fibers in the pectoralis major muscle of chickens contains two high-molecular-weight proteins, i.e., myomesin (Mr, 185,000) and M protein (Mr, 165,000), early in embryonic development (7 days in ovo), even though adult type-I fibers lack M protein. The developmental expression of M protein is unusual in that, from 10 to 14 days in ovo, it is gradually suppressed not only in presumptive type-I fibers but also in presumptive type-II fibers formed from primary-generation myotubes. This latter suppression is transient, as M protein is expressed in all adult type-II fibers derived from both the primary- and second-generation myotubes. Myomesin, on the other hand, is continuously expressed in all myotubes throughout development. This finding shows that myomesin and M protein expression is regulated independently in different myotube populations, and that the suppression of M protein in primary-generation myotubes accounts for the delayed accumulation of M protein during development, as previously revealed by biochemical analysis. Presumptive type-I fibers, which form in the deep portion of the muscle, become concentrated in a narrow band known as the red strip.

Aging↗

Cryoultramicrotomy and immunocytochemistry in the analysis of muscle fine structure.

Cryoultramicrotomy, which avoids the use of harsh fixation procedures, deleterious dehydration and plastic embedding can be combined with immunocytochemistry to determine the ultrastructural localization of cellular proteins. Our attempts to use the cryosectioning technique in combination with immunolabelling to bridge the gap between light and electron microscopic analysis of muscle morphology have enabled us to obtain new information on fibre typing at the ultrastructural level. Furthermore, we have obtained a marked improvement in the resolution of myofibrillar structures by using semithin cryosections for fluorescence microscopy. Data are also presented on correlated light and electron microscope immunocytochemistry of myocardial intermediate filaments confirming the presence of longitudinally oriented intermediate filaments of desmin in the region of the intercalated discs of mammalian cardiac myocytes, whereas elsewhere in the myocyte the bulk of intermediate filaments of desmin is concentrated in the intermyofibrillar space at the level of the Z disc.

Animals↗

Fractionation of mutagenic compounds formed during chlorination of humic water.

Mutagenic compounds formed during chlorination of humic water were fractionated by reversed phase high performance liquid chromatography, size exclusion chromatography and thin layer chromatography, in sequence. Following each chromatographic separation the mutagenic compounds were found in one fraction predominantly. A remarkable purification of active compounds was achieved without serious losses of activity, thus enabling further purification and mass spectrometric studies of the main mutagens.

Chlorine↗

Characterisation of volatile organic compounds in stemwood using solid-phase microextraction.

Solid-phase microextraction (SPME), hydrodistillation and dynamic headspace combined with GC and GC-MS were applied and compared for the analysis of volatile organic compounds (VOCs) from coniferous wood. The SPME conditions (type of fibre, size of wood sample, temperature and exposure time) were optimised, and more than 100 VOCs and semi-volatile compounds extracted and identified from the sapwood and heartwood of Norway spruce (Picea abies). The total number of mono- and sesquiterpenes eluted and identified was similar for the SPME and hydrodistillation methods, but more semi-volatile compounds were released by hydrodistillation. By applying dynamic headspace at room temperature, it was possible to analyse only the most volatile compounds. The qualitative composition of VOCs was similar in spruce sapwood and heartwood, although Z-beta-ocimene occurred only in sapwood while fenchol was present only in heartwood. SPME sampling coupled with GC, applied here to the analysis of VOCs released from stemwood of firs for the first time, is a convenient, sensitive, fast, solvent-free and simple method for the determination of wood volatiles. The technique requires much smaller sample amounts compared with hydrodistillation, and the total amount of VOCs extracted and identified is higher than that obtained by hydrodistillation or dynamic headspace. The relative ratios of the main mono- and sesquiterpenes and -terpenoids were similar using the SPME-GC and hydrodistillation methods.

Organic Chemicals↗

Contact allergy to acid and neutral fractions of rosins. Sensitization experiments in guinea pigs and patch testing in patients.

The allergenicity of two different types of rosins (gum rosin and tall oil rosin) was compared. The rosins were divided into their neutral and acid fractions. The neutral fraction of tall oil rosin failed to induce contact sensitivity in animals tested according to the Guinea pig maximization test method (GPMT). The neutral fraction of gum rosin as well as the two acid fractions gave significant responses. Relatively fewer dermatitis patients reacted to the neutral fraction compared with reactions to the unfractionated gum rosin when patch tested. Fewer reactions to tall oil rosin than to gum rosin (p less than 0.05) were observed. It is concluded that tall oil rosin is less allergenic than gum rosin, which may be due to the absence of allergens in its neutral fraction.

Animals↗