Conglutinin, immunoconglutinins and heterophile antibodies in the sera of apparently healthy moose (Alces alces) and caribou (Rangifer tarandus) in Quebec

Serum samples from apparently healthy wild populations of moose and caribou in the province of Quebec, Canada were screened for the presence of conglutinin (K), immunoconglutinins (IKS) and heterophile antibodies. The conglutinating factor in moose and caribou sera was characterized utilizing the necessity of calcium ions for its reaction with sensitized sheep erythrocytes which had been alexinated with horse complement. The conglutinating substance in these animals did not require calcium ions for its activity. The conglutinating activity in both moose and caribou sera was characterized due to IKS as those present in sheep, dog and rabbit sera. Both moose and caribou had non-agglutinating type of heterophile antibodies. Their titres varied from 0 to 80. None of the animals tested had K in their blood. The titre of IKS varied from 0 to 640 with a mean value of 41 in moose, whereas it varied from 0 to 80 with a mean value of 18 in caribou. About 75% of all the animals in both the groups were positive for IKS. The specificity of IKS was demonstrated by the total removal of its activity on absorption with alexinated cells. Presence of IKS in these animals is suggestive of latent infection(s) possibly of bacterial, viral or parasitic origin.     

Isolation and characterization of a mitochondrial D-amino acid oxidase from Neurospora crassa.

D-Amino acid oxidase (EC 1.4.3.3) activity in homogenates of Neurospora crassa strain SY7A was found to sediment with the mitochondrial fraction. Digitonin fractionation studies on purified mitochondria have indicated a matrix localization of the enzyme. Additionally, a peroxidase (EC 1.11.1.7) activity, which may remove hydrogen peroxide formed as a product of D-amino acid oxidation, was also found in the mitochondrial matrix. Partial purification (20- to 30-fold) of the mitochondrial D-amino acid oxidase was achieved. The enzyme exhibited a pH optimum between 9.0 and 9.2, temperature optimum between 20 and 30 degrees C, and a molecular weight of 118 000 +/- 6000 as determined by gel electrophoresis and 125 000 as determined by gel chromatography.     

Macrobenthic species composition and diversity in the Godthaabsfjord system, SW Greenland

The southwest Greenland coast is made up of large and deep sill fjords. On the shelf, a number of shallow banks separated by deep troughs are located 20–50 km from the coast. We collected three 0.1-m² van Veen grabs at nine stations along a transect spanning from the inner Godthaabsfjord influenced by glaciers, across the shallow Fyllas Bank and out to the slope of the continental shelf at approximately 1,000 m depth. Along this transect, we explored patterns of macrobenthic diversity, species composition, abundance and biomass. The sampled stations were very different in terms of environmental variables, resulting in large differences in species composition primarily related to differences in depth, silt–clay fraction and chl a content of the sediment (BIO-ENV analysis). Habitat differences also reduced species spatial ranges and the majority of species were found at only one (49%) or two (20%) stations and, consequently, species turnover or beta diversity was high and correlated to differences in depth, silt–clay fraction and median sediment grain size. Species richness and diversity were lowest in sites exposed to sediment disturbance: near the glaciers in the inner fjord (physical disturbance by mineral sedimentation) and at selected stations on the shelf (bioturbation by burrowing sand eel). Alpha diversity and richness were only weakly correlated to environmental parameters, indicating that alpha richness and diversity are influenced by several factors or that relationships are non-linear as was found for species richness and silt–clay fraction.     

Improved Flotation Technique for Microscopy of In Situ Soil and Sediment Microorganisms

An improved flotation method for microscopy of in situ soil and sediment microorganisms was developed. Microbial cells were released into gellike flotation films that were stripped from soil and sediment aggregates as these aggregates were submerged in 0.5% solutions of polyvinylpyrrolidone. The use of polyvinylpyrrolidone solutions instead of water facilitated the release of films from saturated samples such as aquifer sediments as well as from typical surface soils. In situ microbial morphological characteristics could then be surveyed rapidly by light microscopy of films stained with acridine orange. This method effectively determined the ranges of morphological diversity in a variety of sample types. It also detected microcolonies and other spatial relationships among microbial cells. Only a small fraction (3.4 to 10.1%) of the microflora was released into the flotation films, but plating and direct evaluations by microscopy showed that this fraction was representative of the total population.     

Metal retention and distribution in the sediment of a constructed wetland for industrial wastewater treatment

A free water surface wetland was built in 2002 to treat wastewater from a tool factory containing metals (Cr, Ni, Zn and Fe), nutrients and organic matter. Until 2006, the last reported period, the wetland retained metals and stored them primarily in the bottom sediment and in the biomass of macrophytes secondarily. The aim of this work was to study metal retention and distribution in the sediment of a constructed wetland for industrial wastewater treatment. Total concentrations and fractions (exchangeable, carbonate-bound, Fe-Mn oxides-bound, organic matter-bound and residual) of metals in sediment were analyzed in this treatment wetland, in order to estimate the fate of metals over time. Metal concentrations were significantly higher in the inlet than in the outlet sediment; concentrations in the latter remained without significant differences throughout the testing period. Metal concentrations and redox potential decreased with depth within the sediment. The lowest metal concentrations and pH and the highest redox values were attained in spring, in agreement with the period of maximum macrophyte growth. Ni and Zn were mainly stored associated with the carbonate fraction; Cr was mainly associated with the Fe-Mn oxides fraction, while Fe was mainly associated with the residual fraction, probably as pyrite. The incoming wastewater composition containing high pH, carbonate, calcium and Fe concentrations favored the observed association in the surface sediment. It would be expected that sediment will continue retaining metals in fractions that will not release them into the water while the chemical and environmental conditions remain unchanged.     

Assessment of factors influencing direct enumeration of viruses within estuarine sediments

Accurate enumeration of viruses within environmental samples is critical for investigations of the ecological role of viruses and viral infection within microbial communities. This report evaluates differences in viral and bacterial direct counts between estuarine sediment samples which were either immediately processed onboard ship or frozen at -20 degrees C and later processed. Viral and bacterial abundances were recorded at three stations spanning the length of the Chesapeake Bay in April and June 2003 within three sediment fractions: pore water (PW), whole sediment (WS), and sediment after pore water removal (AP). No significant difference in viral abundance was apparent between extracts from fresh or frozen sediments. In contrast, bacterial abundance was significantly lower in the samples subjected to freezing. Both bacterial and viral abundance showed significant differences between sediment fractions (PW, WS, or AP) regardless of the fresh or frozen status. Although pore water viral abundance has been used in the past as a measurement of viral abundance in sediments, this fraction accounted for only ca. 5% of the total sediment viral abundance across all samples. The effect of refrigerated storage of sediment viral extracts was also examined and showed that, within the first 2 h, viral abundance decreased ca. 30% in formalin-fixed extracts and 66% in unfixed extracts. Finally, the reliability of direct viral enumeration via epifluorescence microscopy was tested by using DNase treatment of WS extractions. These tests indicated that a large fraction (>86%) of the small SYBR gold fluorescing particles are likely viruses.     

Assessment of Factors Influencing Direct Enumeration of Viruses within Estuarine Sediments

Accurate enumeration of viruses within environmental samples is critical for investigations of the ecological role of viruses and viral infection within microbial communities. This report evaluates differences in viral and bacterial direct counts between estuarine sediment samples which were either immediately processed onboard ship or frozen at −20°C and later processed. Viral and bacterial abundances were recorded at three stations spanning the length of the Chesapeake Bay in April and June 2003 within three sediment fractions: pore water (PW), whole sediment (WS), and sediment after pore water removal (AP). No significant difference in viral abundance was apparent between extracts from fresh or frozen sediments. In contrast, bacterial abundance was significantly lower in the samples subjected to freezing. Both bacterial and viral abundance showed significant differences between sediment fractions (PW, WS, or AP) regardless of the fresh or frozen status. Although pore water viral abundance has been used in the past as a measurement of viral abundance in sediments, this fraction accounted for only ca. 5% of the total sediment viral abundance across all samples. The effect of refrigerated storage of sediment viral extracts was also examined and showed that, within the first 2 h, viral abundance decreased ca. 30% in formalin-fixed extracts and 66% in unfixed extracts. Finally, the reliability of direct viral enumeration via epifluorescence microscopy was tested by using DNase treatment of WS extractions. These tests indicated that a large fraction (>86%) of the small SYBR gold fluorescing particles are likely viruses.     

A test of normalization methods for marine sediment,including a new post-extraction normalization (PEN) technique

Chemical analyses of sediment are used for assessing the ability of sediment to support a healthy benthos (sediment quality) and for determining contaminant source and dispersion in aquatic systems. Total sediment analysis is used for sediment quality assessment, whereas source identification and dispersion requires normalised contaminant data. Normalized contaminant data are obtained by physical fractionation (size-normalization) of the sediment and analyses of a constant size fraction (usually the 62.5 m fraction), whereas elemental normalization uses the total sediment analysis normalized to a conservative element. Elemental normalization is preferable, as it is cheaper and less time consuming than size-normalization techniques. In addition, some contaminants associated with oxides and oxyhydroxides in the coarse fraction are excluded in fine fraction analyses. Five techniques used to normalize sedimentary contaminant data were tested in the current study, including a new post-extraction normalization method where total sediment data are normalized to the residue after digestion, on the assumption that this fraction acts as a diluent only. Results of the tests indicated that simple normalization to the mud fraction provides useful dispersion information, but that the post-extraction normalization method produced a superior indication of source. Limited source and dispersion information was gleamed from the elemental-normalization (Al, Fe) approach, whereas the size-normalization technique provided the clearest indication of source and dispersion. Simple mud normalization and post-extraction normaliaation methods should be considered because only one analysis provides sediment quality, as well as source and dispersion information. However, for detailed information on source and dispersion, size normalization is recommended.     

Subcellular Localization and Differentiation-Induced Redistribution of the Protein Tyrosine Phosphatase PTP-BL in Neuroblastoma Cells

1.In cells of epithelial origin the protein tyrosine phosphatase PTP-BL is predominantly localized at the apical membrane of polarized cells. This large submembranous multidomain PTP is also expressed in cells of neuronal origin. We studied the localization of PTP-BL in mouse neuroblastoma cells utilizing EGFP-tagged versions of the protein. 2. In proliferating Neuro-2a cells, immunofluorescence and immuno-electron microscopy revealed a submembranous FERM domain-dependent localization at cell-cell boundaries for EGFP-PTP-BL. Additionally, significant amounts of EGFP-PTP-BL are located in the cytoplasm as well as in nuclei. Upon serum depletion-induced differentiation of Neuro-2a cells, a partial shift of EGFP-PTP-BL from a cortical localization to cytoskeleton-like F-actin-positive structures is observed. Parallel biochemical studies corroborate this finding and reveal a serum depletion-induced shift of EFGP-PTP-BL from a membrane(-associated) fraction to an NP40-soluble cytoskeletal fraction. 3. Different pools of PTP-BL-containing protein complexes can be discerned in neuronal cells, reflecting distinct molecular microenvironments in which PTP-BL may exert its function.     

25-Hydroxylation of 1 alpha-hydroxyvitamin D-3 in rat and human liver

1 alpha-Hydroxyvitamin D-3 25-hydroxylase activity was measured in subcellular fractions of rat and human liver. The formation of 1,25-dihydroxyvitamin D-3 was determined by high pressure liquid chromatography. In rat liver 1 alpha-hydroxyvitamin D-3 25-hydroxylase activities were found in the purified nuclei, the heavy mitochondrial fraction and the microsomal fraction. The enrichment of 25-hydroxylase activity was highest in the heavy mitochondrial fraction. With this fraction a minimum amount (about 0.5 mg) of protein was required before formation of 1,25-dihydroxyvitamin D-3 could be detected. Above this amount the reaction was linear with amount of protein up to at least 2 mg/ml. The reaction was also linear with time up to 60 min. An apparent Km value of 2 X 10(-5) M was found. The mitochondrial 25-hydroxylase was stimulated by addition of cytosolic protein or bovine serum albumin. The degree of stimulation was dependent on the amount of mitochondrial protein present in the incubation mixture. Maximal stimulation was seen with 0.2 mg/ml of either protein in the presence of 0.5 mg mitochondrial protein. The stimulating effect remained after heating the protein for 5 min at 100 degrees C. The cytosolic protein did not stimulate a reconstituted mitochondrial 1 alpha-hydroxyvitamin D-3 25-hydroxylase. The mitochondrial vitamin D-3 25-hydroxylase was inhibited both by cytosolic protein and by bovine serum albumin. Human liver revealed only one 1 alpha-hydroxyvitamin D-3 25-hydroxylase activity located to the heavy mitochondrial fraction. The results are in agreement with previous studies on the localization of vitamin D-3 25-hydroxylase in rat and human liver. The difference in localization of the 25-hydroxylase between rat and human liver implies that studies on the regulation of the microsomal 25-hydroxylase in rat liver may not be relevant to the situation in human liver.     

UniFrac: a New Phylogenetic Method for Comparing Microbial Communities

We introduce here a new method for computing differences between microbial communities based on phylogenetic information. This method, UniFrac, measures the phylogenetic distance between sets of taxa in a phylogenetic tree as the fraction of the branch length of the tree that leads to descendants from either one environment or the other, but not both. UniFrac can be used to determine whether communities are significantly different, to compare many communities simultaneously using clustering and ordination techniques, and to measure the relative contributions of different factors, such as chemistry and geography, to similarities between samples. We demonstrate the utility of UniFrac by applying it to published 16S rRNA gene libraries from cultured isolates and environmental clones of bacteria in marine sediment, water, and ice. Our results reveal that (i) cultured isolates from ice, water, and sediment resemble each other and environmental clone sequences from sea ice, but not environmental clone sequences from sediment and water; (ii) the geographical location does not correlate strongly with bacterial community differences in ice and sediment from the Arctic and Antarctic; and (iii) bacterial communities differ between terrestrially impacted seawater (whether polar or temperate) and warm oligotrophic seawater, whereas those in individual seawater samples are not more similar to each other than to those in sediment or ice samples. These results illustrate that UniFrac provides a new way of characterizing microbial communities, using the wealth of environmental rRNA sequences, and allows quantitative insight into the factors that underlie the distribution of lineages among environments.     

Distribution of sodium-plus-potassium-stimulated adenosine-triphosphatase activity in isolated nerve-ending particles

1. A rapid method for the isolation of nerve-ending particles from brain is described. This involved the centrifugation of the large-granule fraction over a discontinuous density gradient consisting of 3% (w/v) and 13% (w/v) Ficoll dissolved in 0.32m-sucrose. The results of the biochemical as well as morphological identification of nerve-ending particles are given. 2. Approx. 20% of the (Na(+)+K(+))-stimulated adenosine-triphosphatase activity originally present in the cerebral grey-matter suspension was recovered in the fraction consisting principally of large nerve-ending particles (approx. 1mu in diameter). The activity of the adenosine triphosphatase/mg. of protein in the nerve-ending fraction approximated to that in the small-granule fraction after the treatment with glycol ether diamine-tetra-acetic acid. The conclusion was drawn that the synaptic structure, supposedly the limiting membrane of the nerve-ending particle, is one of the feasible sites of localization of the (Na(+)+K(+))-stimulated adenosine-triphosphatase activity in cerebral tissues. Adenosine triphosphatase in purified cerebral mitochondria was not stimulated by Na(+). 3. No qualitative differences were found between the (Na(+)+K(+))-stimulated adenosine-triphosphatase activities exhibited by the nerve-ending particles and by the cerebral small-granule fraction with respect to pH-dependence, cation requirements and susceptibility to ouabain.     

UniFrac: a new phylogenetic method for comparing microbial communities

We introduce here a new method for computing differences between microbial communities based on phylogenetic information. This method, UniFrac, measures the phylogenetic distance between sets of taxa in a phylogenetic tree as the fraction of the branch length of the tree that leads to descendants from either one environment or the other, but not both. UniFrac can be used to determine whether communities are significantly different, to compare many communities simultaneously using clustering and ordination techniques, and to measure the relative contributions of different factors, such as chemistry and geography, to similarities between samples. We demonstrate the utility of UniFrac by applying it to published 16S rRNA gene libraries from cultured isolates and environmental clones of bacteria in marine sediment, water, and ice. Our results reveal that (i) cultured isolates from ice, water, and sediment resemble each other and environmental clone sequences from sea ice, but not environmental clone sequences from sediment and water; (ii) the geographical location does not correlate strongly with bacterial community differences in ice and sediment from the Arctic and Antarctic; and (iii) bacterial communities differ between terrestrially impacted seawater (whether polar or temperate) and warm oligotrophic seawater, whereas those in individual seawater samples are not more similar to each other than to those in sediment or ice samples. These results illustrate that UniFrac provides a new way of characterizing microbial communities, using the wealth of environmental rRNA sequences, and allows quantitative insight into the factors that underlie the distribution of lineages among environments.     

Studies on the nature of microsome-bound substances involved in the biosynthesis of acidic glycoproteins of guinea-pig serum

1. In further studies on the biosynthesis of components of fraction I, an acidic glycoprotein-containing fraction from guinea-pig serum, an investigation was made of the substances bound to liver microsomes that had earlier been implicated to participate in the synthesis of components of fraction I present in serum. These substances were normally liberated by ultrasonic vibrations. Antisera to subfractions of fraction I were used for characterization. 2. At pH8.6, most of the microsome-bound substances showed electrophoretic mobilities lower than components of fraction I but similar to components of sialic acid-free fraction I. 3. The sialic acid/protein ratios of immune precipitates formed by microsome extracts were similar to those of precipitates formed by sialic acid-free fraction I. 4. On chromatography on Sephadex G-150, most of the microsomal substances were eluted at an essentially similar volume to the main components of fraction I. 5. It was concluded that most of the microsome-bound substances lack sialic acid residues, and, as appreciable degradation of completed molecules is unlikely, these substances appear to be precursors of serum glycoprotein molecules with incomplete prosthetic groups. 6. Evidence was obtained on the submicrosomal localization of incomplete and complete serum glycoprotein molecules.     

High-molecular-weight serum protein complexes differentially promote cell migration and the focal adhesion localization of the urokinase receptor in human glioma cells

The distribution of the urokinase-type plasminogen activator receptor (uPAR) on human glioma cells was examined as a function of culture conditions, using immunofluorescence and immunophotoelectron microscopy. Both uPAR colocalization with focal adhesion proteins and glioma cell motility were maximal in medium containing whole serum or a serum fraction retained by a 500,000 mol wt cutoff centrifugal concentration filter. High motility also took place in medium containing a serum fraction passed by the 500,000 cutoff filter but retained by a 100,000 cutoff filter and in minimal medium containing added vitronectin; however, under these conditions only a small percentage of the otherwise abundant focal adhesions contained colocalized uPAR. Glioma cells in minimal medium with added laminin migrated with a highly elongated morphology but without either classical focal adhesions or well-defined uPAR labeling. In contrast, glioma cells in minimal medium with no additions did not migrate, nor did they adhere well or display defined labeling patterns for focal adhesion proteins or uPAR. The results indicate that high-molecular-weight serum protein complexes promote both uPAR-focal adhesion colocalization and cell migration in glioma cells. However, conditions can be selected in which migration takes place with minimal uPAR-focal adhesion localization, as well as in the absence of apparent focal adhesions.     

Carbon isotope fractionation during acetoclastic methanogenesis by Methanosaeta concilii in culture and a lake sediment

The isotope enrichment factors (epsilon) in Methanosaeta concilii and in a lake sediment, where acetate was consumed only by Methanosaeta spp., were clearly less negative than the epsilon usually observed for Methanosarcina spp. The fraction of methane produced from acetate in the sediment, as determined by using stable isotope signatures, was 10 to 15% lower when the appropriate epsilon of Methanosaeta spp. was used.     

Determination of non-cholesterol sterols in serum and HDL fraction by LC/MS-MS: Significance of matrix-related interferences

BackgroundNon-cholesterol sterols (NCS) are promising biomarkers for estimation of cholesterol homeostasis properties. In addition, determination of NCS in high-density lipoprotein (HDL) fraction (HDL-NCS) could provide information on cholesterol efflux. However, matrix effects interfere in liquid chromatography-mass spectrometry (LC-MS) analysis of NCS, thereby impairing the method sensitivity. The aims of this study were development, optimization and validation of LC-MS method for quantification of NCS in serum and HDL-NCS. Additionally, matrix effect interferences and methods application in individual serum samples were examined.MethodsHDL precipitating reagent was used for HDL isolation. Matrix effect was examined by comparing different surrogates by simple regression analysis. Validation was conducted according to the FDA-ICH guideline. 20 healthy volunteers were recruited for testing of method application.ResultsThe observed matrix effect was 30%, and matrix comparison showed that cholesterol was the dominant contributor to the matrix effect. Cholesterol concentration was adjusted by construction of the calibration curve for serum and HDL fraction (5 mmol/L and 2.5 mmol/L, respectively). The intraand interrun variabilities for NCSs were 4.7-10.3% for serum NCS and 3.6-13.6% for HDLNCS and 4.6-9.5% for serum NCSs and 2.5-9.8% for HDL-NCS, respectively. Recovery studies showed satisfactory results for NCSs: 89.8-113.1% for serum NCS and 85.3-95.8% for HDL-NCS.ConclusionsThe method was successfully developed and optimized. The matrix interference was solved by customising calibration curves for each method and sample type. The measurement of NCS in HDL fraction was proposed for the first time as potentially useful procedure in biomedical researches.     

秋茄(Kandelia candel (L))根系分泌低分子量有机酸及其对重金属生物有效性的影响

应用电感耦合等离子体质谱(ICP-Ms)与高效液相色谱(HPLC)分别对福建漳江口红树林湿地不同土壤结构(砂质与泥质)根际与非根际沉积物中重金属(Cu,Pb,Cd,Zn)含量,以及生长于砂质与泥质滩涂上的红树植物秋茄(Kandelia candel(L)Druce))幼苗根系分泌物中的低分子量有机酸进行测定。在室内模拟秋茄根系分泌的低分子量有机酸,作为重金属提取剂提取沉积物中可溶解态与碳酸盐结合态重金属,并与欧盟标准物质局(BCR)连续提取法相比较,探讨红树根系分泌的低分子量有机酸对红树林沉积物重金属生物有效性的影响。研究结果表明:漳江口红树林泥质沉积物中重金属含量大于砂质沉积物,根际沉积物大于非根际沉积物。两样地沉积物中重金属的赋存形态主要以铁锰氧化物结合态为主,根际沉积物中可交换态与碳酸盐结合态重金属含量均大于非根际沉积物。秋茄根系分泌低分子量有机酸为甲酸,丁酸,苹果酸,柠檬酸,乳酸。不同土壤结构对秋茄根系分泌的苹果酸,柠檬酸,乳酸有显著影响(P<0.05)。以低分子量有机酸作为提取剂对沉积物中可溶解态与碳酸盐结合态重金属的提取率表现为:柠檬酸>混合酸>苹果酸>乳酸>乙酸,低分子量有机酸对红树林沉积物重金属的生物有效性有促进作用。     

On the latency and nature of phenoloxidase present in the left colleterial gland of the cockroach Periplaneta americana

The phenoloxidase system responsible for the sclerotization of cockroach ootheca is found to be present as an inactive form in the left colleterial gland of Periplaneta americana. The supernatant fraction obtained by centrifugation of the milky white secretions contained the inactive phenoloxidase which required both sodium dodecyl sulfate (SDS) and the insoluble sediment for exhibiting enzyme activity. Bovine serum albumin could replace the sediment in the activation process. Proteins separated from the supernatant fraction by molecular sieve chromatography on Sephadex G-25 did not require either albumin or the sediment, but required SDS for exhibiting the phenoloxidase activity. Among the detergents tested, SDS (anionic) and cetylpyridinium chloride (cationic) activated the phenoloxidase, but CHAPS (zwitterionic) or nonionic detergents failed to activate the enzyme. The activation caused by SDS occurred well below the critical micellar concentration of SDS indicating that SDS is causing the activation by binding to the protein and altering its conformation. Chloroform-methanol extracts of vestibulum or right gland could replace SDS confirming the presence of endogenous activator(s) of phenoloxidase system. A variety of exogenously added lipids could activate the latent enzyme, among which linoleate, oleate, laurate, linolenate, phosphatidylethanolamine, and phosphatidylglycerol proved to be the effective activators of the latent phenoloxidase. Partially purified phenoloxidase was found to be extremely labile and lost its activity on a) freezing and thawing, b) dialysis, and c) heating for 10 min at 55 degrees C. It exhibited a pH optimum of 7 and was inhibited drastically by phenylthiourea and diethyldithiocarbamate. It readily oxidized a number of o-diphenols such as 3,4-dihydroxybenzylalcohol, 3,4-dihydroxyphenethyl alcohol, catechol, N-acetyldopamine, N-acetylnorepinephrine, dopa, dopamine, etc., but failed to oxidize both 3,4-dihydroxybenzoic acid and 3,4-dihydroxybenzaldehyde. It neither converted the typical laccase substrate syringaldazine to its quinone methide product, nor oxidized the p-diphenols, hydroquinone and methylhydroquinone. Therefore, the enzyme participating in the quinone tanning of cockroach ootheca appears to be a typical o-diphenol oxidase and not a laccase as previously thought.     

Hartmannella culbertsoni: enzymatic, ultrastructural, and cytochemical characteristics of peroxisomes in a density gradient

Isopycnic density gradient centrifugation techniques demonstrated that catalase (EC 1.11.1.6) and urate oxidase (EC 1.7.3.3) had similar distribution patterns with a peak at equilibrium density 1.22 suggesting that both enzymes were associated with a single population of subcellular particles. Catalase (EC 1.11.1.6) was shown cytochemically to be associated with peroxisomes in the sediment of the catalase-rich fractions. Protein showed a bimodal distribution with a soluble peak at density 1.10 and a particulate peak at density 1.20. The particulate protein peak corresponded to the mitochondrial peak. Acid phosphatase (EC 3.1.3.2) had an equilibrium density of 1.10. Acid phosphatase (EC 3.1.3.2) localization and ultrastructural examination of the acid phosphatase-rich fraction revealed that activity was associated with vacuoles. No primary lysosomes were identified.