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Comparison of cellulose extraction methods for analysis of stable-isotope ratios of carbon and oxygen in plant material.

The Jayme-Wise and diglyme-HCl methods for extracting cellulose from plant material for stable-isotope analysis differ considerably in ease of use, with the latter requiring significantly less time and specialized equipment. However, the diglyme-HCl method leaves a small lignin residue in the crude cellulose that may affect stable-isotope values, whereas alpha-cellulose produced by the Jayme-Wise method is relatively pure. We examined whether adding a bleaching step to the diglyme-HCl method could produce cellulose of comparable purity to alpha-cellulose by comparing the yield, percent carbon, and carbon (delta13C) and oxygen (delta18O) stable isotope ratios of the two celluloses. We tested each method on the wood of five species that differ in ease of delignification, Eucalyptus maculata Hook., E. botryoides Sm., E. resinifera Sm., Pinus pinaster Ait. and Callitris glaucophylla J. Thompson & L.A.S. Johnson, as well as the foliage of C. glaucophylla. For hardwoods such as the eucalypts, the diglyme-HCl method without bleaching produced cellulose with delta13C and delta18O ratios similar to alpha-cellulose. For the softwood, C. glaucophylla, 3 h of bleaching with acidified chlorite following treatment with diglyme-HCl produced cellulose with delta13C and delta18O ratios similar to alpha-cellulose. Bleached and unbleached crude celluloses and alpha-cellulose of P. pinaster were similar in delta18O, but not delta13C. Both types of crude cellulose produced from the foliage of C. glaucophylla had significantly different isotope ratios from alpha-cellulose. Overall, the diglyme-HCl method, with or without bleaching, appears to be a simple, fast method for extracting alpha-cellulose from hardwoods for stable-isotope analyses, but its suitability for softwoods and foliage needs to be evaluated depending on the species.

Carbon Isotopes↗

Effect of cellulose derivatives on alginate microspheres prepared by emulsification.

Generally discrete and spherical calcium alginate microspheres with a high drug encapsulation efficiency were readily prepared by an emulsification process. They were found to release drug rapidly. In the present study, co-polymer in the form of cellulose derivatives was added to sodium alginate in an attempt to modify the drug release profiles of the microspheres. The effects of cellulose derivatives on the morphology and drug encapsulation efficiency of the microspheres were also evaluated. The cellulose derivatives increased the degree of agglomeration of the microspheres. Small and spherical microspheres were produced from cellulose derivatives of low viscosity while larger microspheres which tended to be elongated were produced from cellulose derivatives of high viscosity. The drug encapsulation efficiency and the drug release profiles were influenced by the chemical nature of the cellulose derivative as well as its viscosity. The efficiency of drug encapsulation generally increased while the rate of drug release decreased with increasing viscosity of the cellulose derivatives. Less hydrophilic cellulose derivatives such as methyl cellulose and hydroxypropylmethyl cellulose were found to increase the efficiency of encapsulating sulphaguanidine, while more hydrophilic cellulose derivatives such as hydroxypropyl cellulose and carboxymethyl cellulose had the opposite effect. Among the cellulose derivatives used, only hydroxypropyl cellulose retarded the drug release of the microspheres.

Alginates↗

A clinical study on different cellulosic dialysis membranes.

A controlled clinical study was performed over a period of 8 weeks in two dialysis centres (Rostock, GDR, and Munich, FRG). The aim was to compare a dialysis membrane made of modified cellulose (Hemophan) with classical regenerated cellulose (Cuprophan). Dialysers containing these membranes, together with a cellulose acetate dialyser, were therefore incorporated in a cross-over programme and clinical and biochemical investigations undertaken. The efficacy of the modified cellulosic membrane with respect to urea and creatinine clearance was shown to be comparable to that of regenerated cellulose and cellulose acetate. However, modified cellulose showed an increased clearance for inorganic phosphate, significantly different from that demonstrated by both regenerated cellulose and cellulose acetate. Blood compatibility studies, which included the assessment of C3a activation and the reduction of white blood cell (WBC) and platelet count, clearly demonstrated that in comparison to regenerated cellulose, modified cellulose resulted in significantly less complement activation and WBC reduction. Similarly in comparison to cellulose acetate, modified cellulose showed reduced complement-activating and WBC-reducing properties. The reason for the improved blood compatibility of modified cellulose is not, as was originally assumed, related to binding of complement-inhibiting heparin, but appears instead to be due to the substitution of hydroxyl groups of regenerated cellulose.

Biocompatible Materials↗

A functional cellulose synthase from ascidian epidermis.

Among animals, urochordates (e.g., ascidians) are unique in their ability to biosynthesize cellulose. In ascidians cellulose is synthesized in the epidermis and incorporated into a protective coat know as the tunic. A putative cellulose synthase-like gene was first identified in the genome sequences of the ascidian Ciona intestinalis. We describe here a cellulose synthase gene from the ascidian Ciona savignyi that is expressed in the epidermis. The predicted C. savignyi cellulose synthase amino acid sequence showed conserved features found in all cellulose synthases, including plants, but was most similar to cellulose synthases from bacteria, fungi, and Dictyostelium discoidium. However, unlike other known cellulose synthases, the predicted C. savignyi polypeptide has a degenerate cellulase-like region near the carboxyl-terminal end. An expression construct carrying the C. savignyi cDNA was found to restore cellulose biosynthesis to a cellulose synthase (CelA) minus mutant of Agrobacterium tumefaciens, showing that the predicted protein has cellulose synthase activity. The lack of cellulose biosynthesis in all other groups of metazoans and the similarity of the C. savignyi cellulose synthase to enzymes from cellulose-producing organisms support the hypothesis that the urochordates acquired the cellulose biosynthetic pathway by horizontal transfer.

Agrobacterium tumefaciens↗

Synthetically modified cellulose: an alternative to synthetic membranes for use in haemodialysis?

Renal replacement therapy relies predominantly on the use of cellulose-based membranes. Such membranes have a biocompatibility profile which is inferior to membranes manufactured from synthetic polymers. Synthetically modified cellulose (SMC) is a new, low-flux haemodialysis membrane in which hydroxyl groups have been replaced with benzyl groups. The biocompatibility profile characterized by changes in white cell and platelet counts and the activation of complement components (C3a, C5a and C5b-9) have been studied in vivo and compared with those of cellulose acetate, unmodified cellulose (Cuprophan ) and low-flux polysulphone (Fresenius Polysulfone) in the same group of patients. For SMC, the white cell count at 15 min declined to 65.6% of pretreatment level, compared with 63.8% for the cellulose acetate, 79.6% for low-flux polysulphone and 28.1% for Cuprophan, thereafter returning to pretreatment levels. Both modified cellulose membranes were superior to unmodified cellulose (P = 0.001); the differences between the modified cellulose membranes were not significant statistically. The changes induced by all three cellulose-based membranes exceeded those for low-flux polysulphone (P = 0.001). Associated with the neutropenia was a reduction in platelet count, but this was independent of membrane type. The mean time-averaged concentrations of C3a(des Arg) over 150 min were 1168 ng ml(-1) (SMC), 1030 ng ml(-1) (cellulose acetate), 1297 ng ml(-1) (Cuprophan) and 790 ng ml(-1) (low-flux polysulphone). Equivalent values for C5a(des Arg) were 6.12 (SMC), 2.98 (cellulose acetate), 11.03 (Cuprophan) and 1.33 ng ml(-1) (low-flux polysulphone). C5b-9 values were 385 (SMC), 386 (cellulose acetate), 177 (Cuprophan) and 185 ng ml(-1) (low-flux polysulphone). For each of the complement components the differences between the membranes were significant [P = 0.0009 (C3a(des Arg)), P = 0.0001 (c5a(des Arg) and C5b-9)]. The levels of C5b-9 generated during dialysis also showed a significant positive correlation compared to C5a for all membranes considered as a single group (Pearson's correlation coefficient = 0.870, P = 0.0001). It is concluded that the modification of the cellobiosic unit is a promising approach to improve the biocompatibility profile of cellulose-based membranes. The two different methods of modification lead to similar improvements in biocompatibility compared with unmodified cellulose, but as yet do not match that of low-flux polysulphone.

Biocompatible Materials↗

Influence of cellulose structure on its digestibility in the rat.

The effect of modifying the structural properties of a purified cellulose on its subsequent digestion in the rat was determined. The three structurally unique types of cellulose used in the study were a commercial microcrystalline cellulose, a ball-milled cellulose, and an acid-swollen cellulose. The ball-milled cellulose and the acid-swollen cellulose were prepared from the microcrystalline cellulose starting material. Differences in structure between the types of cellulose were determined by X-ray diffraction measurements and by analysis of their in vitro enzymatic saccharification. The extent of fermentation of each type of cellulose within the rat intestinal tract was determined. Each of the three types of cellulose had a unique structure with respect to its measured crystallinity index and its rate of degradation in vitro by cellulase enzymes. The measured in vivo digestion coefficient for microcrystalline, ball-milled, and acid-swollen cellulose was 8.8, 12.2 and 20.3%, respectively. This represents a significant difference (P less than 0.05) in the extent of fermentation of the different structural types of cellulose within the rat intestine. The results demonstrate that modifying the structural properties of dietary cellulose can affect in vivo response.

Animals↗

Artificial fiber complexes composed of cellulose and guar gum or psyllium may be better sources of soluble fiber for rats than comparable fiber mixtures.

Three fiber complexes composed of cellulose and the same weight of guar gum, cornstarch or psyllium were constructed as three-dimensional networks of cellulose fiber filled with the soluble components. Physiological properties of these fiber complexes were examined in comparison with a mixture of powdered cellulose and the corresponding polysaccharides at a ratio of 1:1 in rats. Another group of rats was fed a fiber-free diet. Body weight gain in the guar gum-cellulose mixture group, but not in guar gum-cellulose complex group, was lower than that of the fiber-free group. The cecal walls in the groups fed the guar gum-cellulose and psyllium-cellulose mixture diets were heavier than those in the corresponding fiber complex-fed groups. Weights of the small intestinal wall and ileal mucosa were also greater in the guar gum-cellulose mixture-fed group than in the guar gum-cellulose complex-fed and the fiber-free diet-fed groups. Fermentable energy in dietary fiber estimated from fecal energy excretion, short-chain fatty acid contents and in vitro short-chain fatty acid production rates in the cecum of the complex-fed groups were similar to those in the fiber mixture-fed groups. However, butyric acid content and production rate in the group fed the guar gum-cellulose complex were markedly higher than in the other groups. The lowering effects of guar gum-cellulose and psyllium-cellulose mixtures on plasma cholesterol concentration tended to be greater than those of guar gum-cellulose and psyllium-cellulose complexes. These findings demonstrate that the artificial fiber complexes supply soluble fibers without increasing mechanical stress to the intestines and decreasing cecal fermentation.

Animals↗

Regulation of cellulose-inducible structures of Clostridium cellulovorans.

Scanning electron microscopy was used to detect ultrastructural protuberances on the cellulolytic anaerobe Clostridium cellulovorans. Numerous ultrastructural protuberances were observed on cellulose-grown cells, but few were detected on glucose-, fructose-, cellobiose-, or carboxymethylcellulose (CMC)-grown cells. Formation of these protuberances was detected within 2 h of incubation in cellulose medium, but 4 h incubation was required before numerous structures were observed on the cells. When a soluble carbohydrate or CMC was mixed with cellulose-grown cells, the ultrastructural protuberances could no longer be detected. In fact, no protuberances were observed within 5 min following the addition of glucose, cellobiose, or methylglucose to cellulose-grown cells. The presence of these protuberances corresponded with the binding of the Bandeiraea simplicifolia BSI-B4 isolectin to the cell. Cellulose-grown cells had a greater level of observable lectin binding than cellobiose-grown cells, and lectin binding was not detected on glucose- or fructose-grown cells. In addition, lectin binding ability was lost by cellulose-grown cells following the addition of glucose, fructose, or methylglucose to the cellulose medium. A cellulose-affinity protein fraction expressing cellulase activity was also detected in cell extracts of cellobiose- or cellulose-grown cultures. However, this protein fraction was not detected in extracts of glucose-grown cultures, and was rapidly lost (within 5 min) following the addition of glucose to cellulose-grown cultures. The ability of C. cellulovorans to adhere to cellulose was also affected by the energy substrate, but not in the same manner as the protuberance formation or the cellulase-containing protein fraction. Rather, cellobiose-, cellulose-, and CMC-grown cultures adhered to cellulose, but this adherence was not affected by addition of glucose to the medium. This is the first report that soluble carbohydrates caused the rapid loss of some cellulose-inducible systems of C. cellulovorans.

Bacterial Adhesion↗

Evaluation of the properties of ethylcellulose-cellulose triacetate microcapsules containing theophylline prepared by different microencapsulation techniques.

Using cellulose triacetate as an added complementary coating material in preparing sustained-release ethylcellulose-cellulose triacetate microcapsules of theophylline, three microencapsulation techniques were investigated. Ethylcellulose-cellulose triacetate composite microcapsules, ethylcellulose-cellulose triacetate dual-walled microcapsules and ethylcellulose microcapsules containing cellulose triacetate matrices were prepared using the non-solvent addition phase separation method. The effects of cellulose triacetate on the release of theophylline from the different ethylcellulose-cellulose triacetate microcapsules were obtained from dissolution studies. The results showed that the release rates of ethylcellulose-cellulose triacetate microcapsules were slower than those obtained from the ethylcellulose microcapsules prepared with similar core to wall ratios. The ethylcellulose microcapsules containing cellulose triacetate matrices had longer release half-times and smaller surface areas than the other capsule preparation. The release patterns of theophylline from the different ethylcellulose-cellulose triacetate microcapsules fitted first-order kinetics. Scanning electron micrographs showed that the surfaces of various ethylcellulose-cellulose triacetate microcapsules were different from those of theophylline, cellulose triacetate matrices of cellulose triacetate microcapsules, and that the surface morphology of ethylcellulose-cellulose triacetate microcapsules was affected by the preparative method.

Capsules↗

Cross-polarization/magic-angle spinning 13C nuclear magnetic resonance study of cellulose I-ethylenediamine complex.

Complete assignments of the cross-polarization/magic-angle spinning (CP/MAS) 13C nuclear magnetic resonance (NMR) spectrum of the cellulose I-ethylenediamine (EDA) complex, which is the intermediate of the reaction from cellulose I to cellulose III(I), were performed. In this paper, we used the 13C-enriched cellulose that was biosynthesized by Acetobacter xylinum ATCC10245 strain from culture medium containing D-(2-13C), D-(3-13C), or D-(5-13C)glucose as a carbon source. After conversion into cellulose I-EDA complex by sufficient EDA treatment, the CP/MAS 13C NMR spectra of the 13C-enriched cellulose I-EDA complexes were measured. As a result, 13C resonance lines of each carbon of the cellulose moiety in the complex appeared as a singlet, suggesting that all glucose residues of the complex are magnetically equivalent. The difference in chemical shifts for each carbon between cellulose I-EDA and cellulose I(alpha), I(beta), and III(I), respectively, suggests that the conformation of the cellulose chains for cellulose I-EDA differs from that for cellulose I(alpha), I(beta), and III(I). In addition, fitting analysis of the 13C spectrum of Valonia cellulose I-EDA complex revealed that the complex contains one EDA molecule per two glucose residues in the cellulose chain.

Journal Article↗

Kinetics of Cellulose Digestion by Fibrobacter succinogenes S85.

Growing cultures of Fibrobacter succinogenes S85 digested cellulose at a rapid rate, but nongrowing cells and cell extracts did not have detectable crystalline cellulase activity. Cells that had been growing exponentially on cellobiose initiated cellulose digestion and succinate production immediately, and cellulose-dependent succinate production could be used as an index of enzyme activity against crystalline cellulose. Cells incubated with cellulose never produced detectable cellobiose, and cells that were preincubated for a short time with thiocellobiose lost their ability to digest cellulose (competitive inhibition [K(infi)] of only 0.2 mg/ml or 0.56 mM). Based on these results, the crystalline cellulases of F. succinogenes were very sensitive to feedback inhibition. Different cellulose sources bound different amounts of Congo red, and the binding capacity was HCl-regenerated cellulose > ball-milled cellulose > Sigmacel > Avicel > filter paper. Congo red binding capacity was highly correlated with the maximum rates of metabolism of cellulose digestion and inversely related to K(infm). Congo red (250 (mu)g/ml) did not inhibit the growth of F. succinogenes S85 on cellobiose, but this concentration of Congo red inhibited the rate of ball-milled cellulose digestion. A Lineweaver-Burk plot of ball-milled cellulose digestion rate versus the amount of cellulose indicated that Congo red was a competitive inhibitor of cellulose digestion (K(infi) was 250 (mu)g/ml).

Journal Article↗

Molecular directionality in cellulose polymorphs.

The recently developed technique of reductive amination, followed by gold labeling, was applied to visualize the reducing ends of cellulose microcrystals from cellulose I, cellulose II, and cellulose III(I). In these crystals, which were also characterized by electron diffraction, the labeling proved that the chains were organized in a parallel fashion in cellulose I from ramie and Valonia and also in cellulose III(I) from Valonia. In microcrystals of cellulose II from mercerized ramie, the labeling method showed that the chains were packed into an antiparallel mode. These results are discussed in terms of the fine structure of cellulose I where neighboring microfibrils of opposite polarity are visualized. The mercerization process whereby cellulose I is converted into cellulose II is therefore best described in terms of an intermingling of the cellulose chains from neighboring microfibrils of opposite polarity. As opposed to the case of mercerization the conversion of cellulose I into cellulose III(I) does not require the participation of neighboring microfibrils since the crystalline domains are converted individually.

Boehmeria↗

Xyloglucan sidechains modulate binding to cellulose during in vitro binding assays as predicted by conformational dynamics simulations.

Cross-links between cellulose microfibrils and xyloglucan (XG) molecules play a major role in defining the structural properties of plant cell walls and the regulation of growth and development of dicotyledonous plants. How these cross-links are established and how they are regulated has yet to be determined. In a previous study, preliminary data were presented which suggested that the different sidechains of XG may play a role in controlling cellulose microfibril-XG interactions. In this study, this question is addressed directly by analyzing to what extent the different sidechains of pea cell wall XG and nasturtium seed storage XG affect their binding to cellulose microfibrils. Of particular importance to this study are the chemical data indicating that pea XG possesses a trisaccharide sidechain, which is not found in nasturtium XG. To this end, conformational dynamic simulations have been used to predict whether oligosaccharides representative of pea and nasturtium XG can adopt a hypothesized cellulose-binding conformation and which of these XGs exhibits a preferential ability to bind cellulose. Extensive analysis of the conformational forms populated during 300 K and high-temperature Monte Carlo simulations established that a planar, sterically accessible, glucan backbone is essential for optimal cellulose-binding. For the trisaccharide sidechain-containing oligosaccharide as found in pea XG, sidechain orientation appeared to regulate the gradual acquisition of this hypothesized cellulose binding conformation. Thus, conformational forms were identified that included the twisted backbone (non-planar) putative solution form of XG, forms in which the trisaccharide sidechain orientation enables increased backbone planarity and steric accessibility, and finally a planar, sterically accessible, backbone. By applying these conformational requirements for cellulose binding, it has been determined that pea XG possesses a two- to threefold occurrence of the cellulose binding conformation than nasturtium XG. Based on this finding, it was predicted that pea XG would bind to cellulose at a higher rate than nasturtium XG. In vitro binding assays showed that pea XG-avicel binding does indeed occur at a twofold higher rate than nasturtium XG-avicel binding. The enhanced ability of pea cell wall XG over nasturtium seed storage XG to associate with cellulose is consistent with a structural role of the former during epicotyl growth where efficient association with cellulose is a requirement. In contrast, the relatively low ability of nasturtium XG to bind cellulose is consistent with the need to enhance the accessibility of this polymer to glycanases during germination. These findings suggest potential roles for XG sidechain substitution, enabling XG to function in a variety of different biological contexts.

Binding Sites↗

Effects of guar gum and cellulose on digesta passage rate, ileal microbial populations, energy and protein digestibility, and performance of grower pigs.

Dietary guar gum and cellulose were studied as purified soluble and insoluble nonstarch polysaccharide (NSP) sources, respectively. A control diet containing 14% cornstarch was formulated. A 7% guar gum, a 7% cellulose, and a 7% guar gum + 7% cellulose diet were formulated by adding the NSP to the control diet at the expense of cornstarch (wt/wt), forming a 2 x 2 factorial arrangement. The objectives were to determine whether guar gum and cellulose altered 1) the passage rate of digesta through the small intestine and total tract; 2) the digestibility of energy and CP, characteristics of the digesta, and microbial populations in the ileum; 3) plasma glucose and ghrelin concentrations; and 4) short-term voluntary feed intake and growth performance of grower pigs. In Exp. 1, 12 pigs (27.0 +/- 1.5 kg of BW) were fitted with an ileal T-cannula and were used in a 2-period change-over design, providing 6 observations per diet. Each period included 18 d: a 12-d acclimation period followed by 2-d feces, 3-d digesta, and 1-d venous blood collection periods. In Exp. 1, guar gum and cellulose slowed the passage rate of digesta through the small intestine by 26 and 18%, respectively (P < 0.05). Guar gum increased total tract retention time of the digesta by 14% (P < 0.05). Guar gum and cellulose increased the viscosity of ileal digesta by 72 and 76%, respectively (P < 0.05). Cellulose reduced ileal energy and CP digestibility (P < 0.05), but guar gum only tended to decrease ileal energy digestibility (P < 0.10). Guar gum and cellulose reduced total tract energy and CP digestibility (P < 0.05). At 60 min after feeding, guar gum decreased plasma glucose by 10% (P < 0.10). Guar gum interacted with cellulose to reduce plasma ghrelin before and after feeding (P < 0.05). Guar gum and cellulose interacted to increase ileal bifidobacteria and enterobacteria (P < 0.05); however, guar gum, but not cellulose, increased ileal clostridia (P < 0.05). In Exp. 2, 20 individually housed grower pigs (5 pigs per diet) had free access to the 4 diets used in Exp. 1 for 14 d. Guar gum and cellulose decreased ADG and reduced ADFI on d 0 to 14 (P < 0.05). In summary, increasing purified NSP in the diet reduced the passage rate of digesta, energy and protein digestibility, and feed intake, but increased ileal bifidobacteria and enterobacteria populations. The effects of cellulose were similar to those of guar gum. In conclusion, monitoring of dietary NSP is a critical factor to achieve predictable digestible nutrient intake and intestinal bacterial populations.

Animal Feed↗

[Effect of oxidized cellulose on fibrin formation and blood platelets].

BACKGROUND: Haemostyptic materials initiate and hasten blood clotting at the site of their application. The properties of haemostyptic materials are used for treatment of capillary and parenchymatous haemorrhage along with surgical treatment. Celluloses one of the biopolymers studied for a long time, suitable because of its biocompatibility and non-toxicity. METHODS AND RESULTS: In the submitted study the authors used microdispersed calcium-sodium salt of oxidized cellulose which is formed by oxidation of cellulose in position C6 (patent Alltracel Pharmaceuticals). The authors investigated the effect of oxidized cellulose on fibrin formation and platelets. Using the optic method of the surface plasmon resonance they investigated the initial stage of interaction between fibrinogen and oxidized cellulose. Oxidized cellulose retards and reduces the interaction of the immobilized fibrin monomer with fibrinogen. Fibrin formation was investigated spectrophotometrically at 350 nm. In the presence of cellulose the period of formation of fibrin gel was prolonged and its turbidity increased, depending on the concentration of the cellulose used. The platelet activation by cellulose was assessed by measuring the released serotonin. For the activation of platelets by cellulose the presence of plasma is necessary, rinsed platelets were not activated by cellulose. It was revealed that direct, interaction of rinsed platelets or fibrinogen with cellulose plays a secondary role. CONCLUSIONS: These data and the retarded activation of platelets in plasma with factor XII deficiency indicate that due to negatively charged oxidized cellulose probably activation of the contact coagulation system occurs and this leads to the activation of platelets and fibrin formation.

Blood Platelets↗

Characterization of a novel cellulose synthesis inhibitor.

The physiological effects of an experimental herbicide and cellulose synthesis inhibitor, N2-(1-ethyl-3-phenylpropyl)-6-(1-fluoro-1-methylethyl)-1,3,5-triazine-2,4-diamine, called AE F150944, are described. In the aminotriazine molecular class, AE F150944 is structurally distinct from other known cellulose synthesis inhibitors. It specifically inhibits crystalline cellulose synthesis in plants without affecting other processes that were tested. The effects of AE F150944 on dicotyledonous plants were tested on cultured mesophyll cells of Zinnia elegans L. cv. Envy, which can be selectively induced to expand via primary wall synthesis or to differentiate into tracheary elements via secondary wall synthesis. The IC50 values during primary and secondary wall synthesis in Z. elegans were 3.91 x 10(-8) M and 3.67 x 10(-9) M, respectively. The IC50 in suspension cultures of the monocot Sorghum halapense (L.) Pers., which were dividing and synthesizing primary walls, was 1.67 x 10(-10) M. At maximally inhibitory concentrations, 18-33% residual crystalline cellulose synthesis activity remained, with the most residual activity observed during primary wall synthesis in Z. elegans. Addition to Z. elegans cells of two other cellulose synthesis inhibitors, 1 microM 2,6-dichlorobenzonitrile and isoxaben, along with AE F150944 did not eliminate the residual cellulose synthesis, indicating little synergy between the three inhibitors. In differentiating tracheary elements, AE F150944 inhibited the deposition of detectable cellulose into patterned secondary wall thickenings, which was correlated with delocalization of lignin as described previously for 2, 6-dichlorobenzonitrile. Freeze-fracture electron microscopy showed that the plasma membrane below the patterned thickenings of AE F150944-treated tracheary elements was depleted of cellulose-synthase-containing rosettes, which appeared to be inserted intact into the plasma membrane followed by their rapid disaggregation. AE F150944 also inhibited cellulose-dependent growth in the rosette-containing alga, Spirogyra pratensis, but it did not inhibit cellulose synthesis in Acetobacter xylinum or Dictyostelium discoideum, both of which synthesize cellulose via linear terminal complexes. Therefore, AE F150944 may inhibit crystalline cellulose synthesis by destabilizing plasma membrane rosettes.

Asteraceae↗

A fish hepatoma cell line (PLHC-1) as a tool to study cytotoxicity and CYP1A induction properties of cellulose and wood chip extracts.

Cytotoxicity and CYP1A induction properties of celluloses and wood chips were studied with a teleost liver cell line, PLHC-1. Cells were exposed to acetone extracts of celluloses produced using new bleaching techniques (elemental chlorine free, ECF; totally chlorine free, TCF) in two sulphate mills or without any bleaching (unbleached, UB) in a sulphite mill. In another set of exposures, celluloses (ECF and TCF bleached) and wood chips (from pine and birch) were collected from a sulphate mill, extracted with acetone, and the extracts used to treat the cells. After exposure, O-deethylation of 7-ethoxyresorufin (EROD, a measure of cytochrome P4501A (CYP1A) catalytic activity), and total protein content, a measure of cytotoxicity, were assayed. The presence of the CYP1A protein in the exposed cells was assessed by immunoblotting. The cellulose and wood chip extracts were able to cause both cytotoxicity and EROD induction in the PLHC-1 cells. In the exposures conducted with the material from three different mills, the celluloses made of birch were more cytotoxic and more potent inducers of EROD activity than were the celluloses of pine. Further, UB celluloses increased EROD activity and caused cytotoxicity at lower doses than material bleached with modern bleaching techniques. In the exposures made with material from one single mill, there were no clear trends between the celluloses made of pine or birch. Wood chips of pine, however, were more cytotoxic than wood chips of birch. Especially with pine wood chips, cytotoxicity interfered with the induction of EROD activity, thus complicating the evaluation of CYP1A induction. CYP1A protein content was not detected in cells exposed to extracts of celluloses or wood chips, possibly due to low amounts of protein available for the assay. Wood and pulp processing, like bleaching, may change the chemical composition of the raw material in a way that reduces the potency for biological effects of the final product, cellulose. This could explain why both UB celluloses and wood chips were more potent in the cells than ECF or TCF bleached celluloses. In this study the PLHC-1 cell line showed its potential for use in evaluating the biological activity existing in pulp and paper mill products and raw materials. The identity and source of the compounds that were able to affect the PLHC-1 cell line remain to be determined.

Animals↗

Surface forces measurements of spin-coated cellulose thin films with different crystallinity.

A systematic study of the surface forces between a cellulose sphere and cellulose thin films of varying crystallinity has been conducted as a function of ionic strength and pH. Semicrystalline cellulose II surfaces and amorphous cellulose films were prepared by spin-coating of the precursor cellulose solutions onto oxidized silicon wafers before regeneration in water. Crystalline cellulose I surfaces were prepared by spin-coating wafers with aqueous suspensions of sulfate-stabilized cellulose I nanocrystals. These preparation methods produced thin, smooth films suitable for surface forces measurements. The interaction with the cellulose I was monotonically repulsive at pH 3.5, 5.8, and 8.5 and at 0.1, 1, and 10 mM ionic strengths. This was attributed to the presence of strongly ionizable sulfur-containing groups on the cellulose nanocrystal surfaces. The amorphous film typically showed a steric interaction up to 100 nm away from the interface that was independent of the solution conditions. A range of surface forces were successfully measured on the semicrystalline cellulose II films; attractive and repulsive regimes were observed, depending on pH and ionic strength, and were interpreted in terms of van der Waals and electrostatic interactions. Clearly, the forces acting near cellulose surfaces are very dependent on the way the cellulose surface has been prepared.

Cellulose↗