Spatial variability of soil properties under Pinus canariensis canopy in two contrasting soil textures

The aim of this study was to investigate how the biomass of extramatrical mycorrhizal mycelia (EMM) is influenced by the addition of different phosphorus (P), potassium (K) and PK fertilizers in peatland forests with variable P and K availability. Four fertilizers were used: apatite, biotite, Rauta-PK? (apatite and ferrosulphate) and a test fertilizer (50% apatite and 50% recycled iron phosphate). Forest plots with four different types of nutrient balance (deficient P and deficient K, deficient P and sufficient K, sufficient P and deficient K, and sufficient P and sufficient K) were studied. The effects on EMM biomass and ectomycorrhizal (EM) biomass in roots were estimated by ergosterol and phospholipid fatty acid (PLFA) analysis using in-growth mesh bags. Nutrients and rare-earth elements in EM roots surrounding the mesh bags were quantified and used as indicators of nutrient transport by the EMM in the mesh bags. The biomass of EMM was enhanced by P, K and PK deficiency of the trees, and EM fungal biomass in the roots was increased by P deficiency. The test fertilizer enhanced EMM biomass in all the plots studied, whereas the other fertilizers did not have any significant effect. No significant interactions between the P and K availability of host and mycelial fertilizer response could be detected. Deficiency of P or K or both in needles did not affect the concentrations of rare-earth elements in the tree roots. Earlier results from laboratory experiments have shown reduced carbon allocation to EM fungi under K deficiency, but this was not the case in these mature forests. Instead, we observed increased EMM biomass in response to both P and K deficiency.     

Fine-scale distribution of pine ectomycorrhizas and their extramatrical mycelium

In order to clarify the functional role of individual ectomycorrhizal (EcM) fungal species in the field, we need to relate their abundance and distribution as mycorrhizas to their abundance and distribution as extramatrical mycelium (EMM). We divided each of four 20 cm x 20 cm x 2 cm slices of pine forest soil into 100 cubes of 2 cm x 2 cm. For each cube, ectomycorrhizas were identified and the presence of EMM of the EcM fungi recorded as ectomycorrhizas was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis of ITS rDNA. Ectomycorrhizas and EMM of seven EcM species were mapped. Spatial segregation of mycorrhizas and EMM was evident and some species produced their EMM in different soil layers from their mycorrhizas. The spatial relationship between mycorrhizas and their EMM generally conformed to their reported exploration types, but EMM of smooth types (e.g. Lactarius rufus) was more frequent than expected. Different EcM fungi foraged at different spatial scales.     

The production and turnover of extramatrical mycelium of ectomycorrhizal fungi in forest soils: role in carbon cycling

There is growing evidence of the importance of extramatrical mycelium (EMM) of mycorrhizal fungi in carbon (C) cycling in ecosystems. However, our understanding has until recently been mainly based on laboratory experiments, and knowledge of such basic parameters as variations in mycelial production, standing biomass and turnover as well as the regulatory mechanisms behind such variations in forest soils is limited. Presently, the production of EMM by ectomycorrhizal (EM) fungi has been estimated at ~140 different forest sites to be up to several hundreds of kg per ha per year, but the published data are biased towards Picea abies in Scandinavia. Little is known about the standing biomass and turnover of EMM in other systems, and its influence on the C stored or lost from soils. Here, focussing on ectomycorrhizas, we discuss the factors that regulate the production and turnover of EMM and its role in soil C dynamics, identifying important gaps in this knowledge. C availability seems to be the key factor determining EMM production and possibly its standing biomass in forests but direct effects of mineral nutrient availability on the EMM can be important. There is great uncertainty about the rate of turnover of EMM. There is increasing evidence that residues of EM fungi play a major role in the formation of stable N and C in SOM, which highlights the need to include mycorrhizal effects in models of global soil C stores.     

A Monte Carlo study of the inferential properties of three methods of shape comparison

Three inferential morphometric methods, Euclidean distance matrix analysis (EDMA), Bookstein's edge-matching method (EMM), and the Procrustes method, were applied to facial landmark data. A Monte Carlo simulation was conducted with three sample sizes, ranging from n = 10 to 50, to assess type I error rates and the power of the tests to detect group differences for two- and three-dimensional representations of forms. Type I error rates for EMM were at or below nominal levels in both two and three dimensions. Procrustes in 2D and EDMA in 2D and 3D produced inflated type I error rates in all conditions, but approached acceptable levels with moderate cell sizes. Procrustes maintained error rates below the nominal levels in 2D. The power of EMM was high compared with the other methods in both 2D and 3D, but, conflicting EMM decisions were provided depending on which pair (2D) or triad (3D) of landmarks were selected as reference points. EDMA and Procrustes were more powerful in 2D data than for 3D data. Interpretation of these results must take into account that the data used in this simulation were selected because they represent real data that might have been collected during a study or experiment. These data had characteristics which violated assumptions central to the methods here with unequal variances about landmarks, correlated errors, and correlated landmark locations; therefore these results may not generalize to all conditions, such as cases with no violations of assumptions. This simulation demonstrates, however, limitations of each procedure that should be considered when making inferences about shape comparisons. © 1996 Wiley-Liss, Inc.     

Non-specificity of Laboratory Test for Diagnosis of Multiple Sclerosis

It has been claimed that the inhibiting effect of linoleic acid on the macrophage electrophoretic mobility test provides a specific laboratory method for the diagnosis of multiple sclerosis (M.S.) and may also enable susceptible relatives of M.S. patients to be identified. Three trials of the method under double-blind conditions have failed to confirm that the test is diagnostically useful.     

A method for lake ecosystem health assessment: an Ecological Modeling Method (EMM) and its application

Ecosystem health is a newly proposed concept that sets new goals for environmental management. Its definition, indexing and assessment methods are still being perfected. An Ecological Modeling Method (EMM) for lake ecosystem health assessment is proposed in this paper. The EMM's procedures are: (1) to analyze the ecosystem structure of a lake in order to determine the structure and complexity of the lake's ecological model; (2) to develop a model having ecological health indicators, by designing a conceptual diagram, establishing model equations, estimating model parameters and being integrated with ecological indicators; (3) to compare the simulated and observed values of important state variables and process rates (i.e. model calibration) in order to evaluate the applicability of the model to lake ecosystem health assessment; (4) to calculate ecosystem health indicators based on the developed model; and (5) to assess lake ecosystem health according to the values of the ecosystem health indicators. The EMM was applied, as a case study, to the ecosystem health assessment of a eutrophic Chinese lake (Lake Chao) between April 1987 and March 1988. A relative order of health states from poor to good was determined as follows: August–October 1987 > April–May 1987 > June–July 1987 > November–December 1987 > January–March 1988. These results compared quite favourably with the actual current conditions at Lake Chao. The EMM method, therefore, was suitable in assessing lake ecosystem health at Lake Chao.     

Complex effects of mammalian grazing on extramatrical mycelial biomass in the Scandes forest‐tundra ecotone

Mycorrhizal associations are widespread in high‐latitude ecosystems and are potentially of great importance for global carbon dynamics. Although large herbivores play a key part in shaping subarctic plant communities, their impact on mycorrhizal dynamics is largely unknown. We measured extramatrical mycelial (EMM) biomass during one growing season in 16‐year‐old herbivore exclosures and unenclosed control plots (ambient), at three mountain birch forests and two shrub heath sites, in the Scandes forest‐tundra ecotone. We also used high‐throughput amplicon sequencing for taxonomic identification to investigate differences in fungal species composition. At the birch forest sites, EMM biomass was significantly higher in exclosures (1.36 ± 0.43 g C/m²) than in ambient conditions (0.66 ± 0.17 g C/m²) and was positively influenced by soil thawing degree‐days. At the shrub heath sites, there was no significant effect on EMM biomass (exclosures: 0.72 ± 0.09 g C/m²; ambient plots: 1.43 ± 0.94). However, EMM biomass was negatively related to Betula nana abundance, which was greater in exclosures, suggesting that grazing affected EMM biomass positively. We found no significant treatment effects on fungal diversity but the most abundant ectomycorrhizal lineage/cortinarius, showed a near‐significant positive effect of herbivore exclusion (p = .08), indicating that herbivory also affects fungal community composition. These results suggest that herbivory can influence fungal biomass in highly context‐dependent ways in subarctic ecosystems. Considering the importance of root‐associated fungi for ecosystem carbon balance, these findings could have far‐reaching implications.     

Enhanced cellulase production through random mutagenesis of Talaromyces pinophilus OPC4-1 and fermentation optimization

Reducing cellulase cost remains a major challenge for lignocellulose to fuel and chemical industries. In this study, mutants of a novel wild-type cellulolytic fungal strain Talaromyces pinophilus OPC4-1 were developed by consecutive UV irradiation, N-methyl-N`-nitro-N-nitrosoguanidine (NTG) and ethylmethane sulfonate (EMS) treatment. A potential mutant EMM was obtained and displayed enhanced cellulase production. Using Solka Floc cellulose as the substrate, through fed-batch fermentation, mutant strain T. pinophilus EMM generated crude enzymes with an FPase activity of 27.0 IU/mL and yield of 900 IU/g substrate. When corncob powder was used, strain EMM produced crude enzymes with an FPase activity of 7.3 IU/mL and yield of 243.3 IU/g substrate. In addition, EMM crude enzymes contained 29.2 and 16.3 IU/mL β-glucosidase on Solka Floc cellulose and corncob power, respectively. The crude enzymes consequently displayed strong biomass hydrolysis performance. For corncob hydrolysis, without supplement of any commercial enzymes, glucose yields of 591.7 and 548.6 mg/g biomass were obtained using enzymes produced from Solka Floc cellulose and corncob powder, respectively. It was 553.9 mg/g biomass using the commercial enzyme mixture of Celluclast 1.5 L and Novozyme 188. Strain T. pinophilus EMM was therefore a potential fungus for on-site enzyme production in biorefinery processes.     

Biomass and compositional responses of ectomycorrhizal fungal hyphae to elevated CO2 and nitrogen fertilization

The extramatrical mycelia (EMM) of ectomycorrhizal fungi make up a large proportion of the microbial diversity and biomass in temperate forest soils. Thus, their response to elevated CO(2) can have large effects on plant nutrient acquisition and carbon movement through forests. Here, the effects of CO(2) and nitrogen (N) fertilization on EMM biomass and community structure in Pinus taeda forest plots were examined using sand-filled mesh bags buried in the field, the contents of which were analyzed by phospholipid fatty acid (PLFA) and DNA sequencing. A total of 2138 sequences comprising 295 taxa were recovered; most (83.5%) were from ectomycorrhizal fungal taxa. No biomass increase was detected in elevated CO(2) plots relative to control plots, but individual taxa responded to both CO(2) and N fertilization, four of the six most abundant taxa were less frequent in N-fertilized plots. Thelephoroid and athelioid taxa were both frequent and abundant as EMM, and thelephoroid richness was extremely high. Russula and Cortinariaceae taxa were less abundant and boletoid taxa were more abundant as EMM relative to ectomycorrhizas. The EMM community, sampled across seasons and years, was dynamic with a high degree of interspecific variation in response to CO(2) enrichment and N fertilization.     

Enzyme-Linked Lectinsorbent Assay Measures N-Acetyl-D-Glucosamine in Matrix of Biofilm Produced by Staphylococcus epidermidis

An enzyme-linked lectinsorbent assay (ELLA) was developed for quantification of in situ biofilm produced by Staphylococcus epidermidis in polystyrene 96-well tissue culture plates with phosphatase-labeled wheat germ agglutinin (WGA) as a specific probe for the GlcNAcβ-1,4 _n component of exocellular matrix material (EMM) that is responsible for intercellular adhesion and accumulation. The ELLA and the modified Christensen dye assay were used to test 13 laboratory strains of coagulase-negative staphylococci and 10 clinical isolates of S. epidermidis. Four biofilm-positive laboratory strains of S. epidermidis were positive by both tests, and six biofilm-negative strains were negative by both. One strain of S. hemolyticus was positive by the ELLA only. Two of the 10 clinical isolates of S. epidermidis were positive by both assays, two were negative by both, and the remaining were positive by the ELLA only. The ELLA was objective, reproducible, specific, sensitive, and useful for screening strains for their capacity to adhere to plastic, produce EMM, and form biofilm. Received: 15 February 1997 / Accepted: 16 April 1997     

Multimodality infarct identification for optimal image-guided intramyocardial cell injections

BackgroundIntramyocardial cell injections in the context of cardiac regenerative therapy can currently be performed using electromechanical mapping (EMM) provided by the NOGA®XP catheter injection system. The gold standard technique to determine infarct size and location, however, is late gadolinium enhanced magnetic resonance imaging (LGE-MRI). In this article we describe a practical and accurate technique to co-register LGE-MRI and NOGA®XP datasets during the injection procedures to ultimately perform image-guided injections to the border zone of the infarct determined by LGE-MRI.ResultsImage registration was successful in all datasets, and resulted in a mean registration error of 3.22 ± 1.86 mm between the MRI surface mesh and EMM points. Visual assessment revealed that the locations and the transmural extent of the infarctions measured by LGE-MRI only partly overlap with the infarct areas identified by the EMM parameters.ConclusionsThe 3D CartBox image registration toolbox enables registration of EMM on pre-procedurally acquired MRI during the catheter injection procedure. This allows the operator to perform real-time image-guided cell injections into the border zone of the infarct as assessed by LGE-MRI. The 3D CartBox thereby enables, for the first time, standardisation of the injection location for cardiac regenerative therapy.

Electronic supplementary material

The online version of this article (doi:10.1007/s12471-014-0604-2) contains supplementary material, which is available to authorized users.     

Estimating human long bone cross-sectional geometric properties: a comparison of noninvasive methods

Cross-sectional properties (areas, second moments of area) have been used extensively for reconstructing the mechanical loading history of long bone shafts. In the absence of a fortuitous break or available computed tomography (CT) facilities, the endosteal and/or periosteal boundaries of a bone may be approximated using alternative noninvasive methods. The present study tests whether cross-sectional geometric properties of human lower limb bones can be adequately estimated using two such techniques: the ellipse model method (EMM), which uses biplanar radiography alone, and the latex cast method (LCM), which involves molding of the subperiosteal contour in combination with biplanar radiography to estimate the contour of the medullary canal. Results of both methods are compared with "true" cross-sectional properties calculated by direct sectioning. The study sample includes matched femora and tibiae of 50 Pecos Pueblo Amerindians. Bone areas and second moments of area were calculated for the midshaft femur and tibia and proximal femoral diaphysis in each individual. Percent differences between methods were derived to evaluate directional (systematic) and absolute (random) error. Multiple regression was also used to investigate the sources of error associated with each method. The results indicate that while the LCM shows generally good correspondence to the true cross-sectional properties, the EMM generally overestimates true parameters. Regression equations are provided to correct this overestimation, and, when applied to another sample, are shown to significantly improve estimates for the femoral midshaft, although corrections are less successful for the other section locations. Our results suggest that the LCM is an adequate substitute for estimating cross-sectional properties when direct sectioning and CT are not feasible. The EMM is a reasonable alternative, although the bias inherent in the method should be corrected if possible, especially when the results of the study are to be compared with data collected using different methods.     

Wood ash application in a managed Norway spruce plantation did not affect ectomycorrhizal diversity or N retention capacity

Ectomycorrhizal (ECM) fungi are key players in N cycling in coniferous forests, and forest management such as application of wood ash can affect their functionality. The aim of this study was to determine the effects of wood ash application on ECM fungal mycelial production, capacity to retain N, diversity and community composition. In-growth mesh bags were installed in control and treated plots. After 6 months, ¹⁵N labeled ammonium and nitrate were applied into the mesh bags, and 24 h later extramatrical mycelium (EMM) was extracted and analyzed. Wood ash had no effects on EMM in-growth, N retention capacity, diversity or community composition. In contrast, there were significant seasonal differences in the amount of EMM produced. These results demonstrate that applying up to 6 t ha⁻¹ of wood ash in this type of plantation forest is a safe management practice that does not increase N leaching or negatively affect ECM fungi.     

Effects of Soil Components and Liming Effect of CCA-Wood Ash upon Leaching of Cu,Cr, and As from CCA-Wood Ash in Ultisol Soil

The burning of wood that has been chemically treated with chromated copper arsenate (CCA) produces an ash containing high concentrations of copper, chromium, and arsenic (CCA-metals). The rainwater-leaching of these metals from burn sites can produce increased soil and water contamination. Soil systems have varying natural abilities to retard leaching and they also impact metals speciation and toxicity through sorption, conversion, and sedimentation-related mechanisms. Recent regulations restricting the use of CCA-treatment have resulted in increased quantities of CCA-treated lumber entering the waste stream, making studies of metals leaching from CCA-wood ash and soil/CCA-wood ash systems important areas of investigation.

Wood ash composition, soil composition, and CCA-metals speciation are all important factors determining the degree of the metal mobility in a soil system containing metals leached from CCA-wood ash. The CCA-metals composition of CCA-wood ash was determined by analytical methods. Both pH and batch leaching studies were used to postulate mobility mechanisms within the CCA-wood ash/soil system. The contrasting effects of untreated-wood ash and CCA-wood ash on soil components are presented in order to assess the potential for enhancement of immobilization mechanisms that increase the soil system retardance of CCA-metals mobility.

Results of this investigation show that the Ultisol test soil retards the mobility of As and Cr compared to CCA-wood ash alone, while Cu mobility is increased in the presence of the Ultisol test soil. Experimental results show that the alkalinity of a CCA-wood ash/Ultisol soil system is lower than that of an untreated-wood ash/Ultisol soil system. This indicates a difference in chemical composition and the potential consumption of hydroxyl ions during treatment by the retardance mechanisms affecting Cr and As mobility. Therefore, the study of these contaminated soil systems and the potential enhancement of immobilization mechanisms are important areas of investigation.     

ENU mutagenesis in the mouse electrophoretic specific-locus test. 2. Mutational studies of mature oocytes.

Experiments were conducted using the biochemical specific-locus test to assess the mutagenicity of N-ethyl-N-nitrosourea (ENU) in mature oocytes of mice. C57Bl/6J females were treated with 100 mg/kg ENU by intraperitoneal injection and mated to untreated DBA males for 1 week following treatment. 1447 progeny were screened for evidence of mutations affecting the electrophoretic mobility of 32 different proteins; two mutants were detected by electrophoretic analyses. These results provide evidence that ENU is a germ-cell mutagen in mouse mature oocytes, although the frequency of mutants is somewhat lower than that obtained from spermatogonia treated with the same dose.     

Effects of intradistrict school mobility and high student turnover rates on early reading achievement

The primary aim of this study was to assess the relations between concurrent, cumulative, and contextual intradistrict school mobility and early reading achievement. Longitudinal administrative school records were used for an entire cohort of students in a large urban district from first through third grade. Findings indicated that students with a concurrent intradistrict school move had lower reading achievement scores at the end of each grade compared to children who did not change schools. Cumulative intradistrict school mobility was also associated with poor reading achievement by the end of third grade. Students enrolled in schools with high turnover rates demonstrated worse reading achievement after accounting for individual school mobility experiences, and this effect worsened as children reached third grade.     

Comparison of protein binding to DNA in vivo and in vitro: defining an effective intracellular target.

We have quantitatively evaluated the affinity of a set of target sites for the integration host factor (IHF) protein of Escherichia coli by their performance as competitors in an electrophoretic mobility shift assay. We also determined how well each of these sites is filled by IHF in vivo. The data show that several natural sites have an affinity not much greater than that required for intracellular occupancy. The data also indicate that very little of the IHF in a cell is present as free protein available for binding, suggesting that binding to non-specific targets dominates the operation of this system. The correlation between in vitro affinity and in vivo occupancy provides a ready means to assess the likely physiological significance of putative IHF sites. It also provides a general method to assess the importance of non-specific interactions by DNA binding proteins inside a cell.     

Exploration type-specific standard values of extramatrical mycelium – a step towards quantifying ectomycorrhizal space occupation and biomass in natural soil

The present study aims at providing standard values for the exploration type (ET)-specific quantification of extramatrical mycelium (EMM) of ectomycorrhizal fungi applicable to ecological field studies. These values were established from mycelial systems of ectomycorrhizae (ECM) synthesized in rhizotrons with near-to-natural peat substrate. Based on image analysis, the “Specific Potential Mycelial Space Occupation” (sPMSO), i.e. the ET-specific complete area that is covered by the EMM systems (mm² cm⁻¹ ECM⁻¹), and the “Specific Actual Mycelial Space Occupation” (sAMSO), i.e. the projection area of mycelial systems (mm² cm⁻¹ ECM⁻¹), were analyzed as an extension of a previously described approach. The “Specific Extramatrical Mycelial Length” (sEML) [m cm⁻¹ ECM⁻¹] and the “Specific Extramatrical Mycelial Biomass” (sEMB) (μg cm⁻¹ ECM⁻¹) were calculated for each of the ET via the proportion of hyphal projected area, hyphal length and biomass, the latter two being derived from previous measurements on Piloderma croceum, a “Medium-Distance” (MD)-ET. Both sPMSO and sAMSO were highest for the “Long-Distance” (LD)-ET, whereas those of the “Short-Distance” (SD)-ET and MD-ET were similar, although showing high variation. In contrast, mycelial density per occupied area of the MD-ET was twice as high as that of the LD-ET. Proportional to the sAMSO, the EMM length and biomass differed considerably between the three ET with values of the MD-ET being 1.9 times higher than those of SD-ET, and those of the LD-ET being 15 times higher than those of the SD-ET. These standards in relation to ECM length may ease quantification of mycelial space occupation and biomass in a relatively simple way. Thereby, the ET-specific contribution of EMM can be distinguished—also of non-cultivable species—and up-scaling to large-scale estimation of cost/benefit relations is possible.     

Site fertility and soil water-table level affect fungal biomass production and community composition in boreal peatland forests

A substantial amount of below-ground carbon (C) is suggested to be associated with fungi, which may significantly affect the soil C balance in forested ecosystems. Ergosterol from in-growth mesh bags and litterbags was used to estimate fungal biomass production and community composition in drained peatland forests with differing fertility. Extramatrical mycelia (EMM) biomass production was generally higher in the nutrient-poor site, increased with deeper water table level and decreased along the length of the recovery time. EMM biomass production was of the same magnitude as in mineral-soil forests. Saprotrophic fungal biomass production was higher in the nutrient-rich site. Both ectomycorrhizal (ECM) and saprotrophic fungal community composition changed according to site fertility and water table level. ECM fungal community composition with different exploration types may explain the differences in fungal biomass production between peatland forests. Melanin-rich Hyaloscypha may indicate decreased turnover of biomass in nutrient-rich young peatland forest. Genera Lactarius and Laccaria may be important in nutrient rich and Piloderma in the nutrient-poor conditions, respectively. Furthermore, Paxillus involutus and Cortinarius sp. may be important generalists in all sites and responsible for EMM biomass production during the first summer months. Saprotrophs showed a functionally more diverse fungal community in the nutrient-rich site.     

Diagnostic and economic evaluation of new biomarkersfor Alzheimer¿s disease: the research protocol of a prospective cohort study

ABSTRACT: BACKGROUND: New research criteria for the diagnosis of Alzheimer's disease (AD) have recently been developed to enable an early diagnosis of AD pathophysiologyby relying on emerging biomarkers. To enable efficient allocation of health care resources, evidence is needed to support decision makers on the adoption of emerging biomarkers in clinical practice. The research goals are to 1) assess the diagnostic test accuracy of current clinical diagnostic work-up and emerging biomarkers in MRI, PET and CSF, 2) perform a cost-consequence analysis and 3) assess long-term cost-effectiveness by an economic model.Methods/designIn a cohort design 223 consecutive patients suspected of having a primary neurodegenerative disease are approached in four academic memory clinics and followed for two years. Clinical data and data on quality of life, costs and emerging biomarkers are gathered.Diagnostic test accuracy is determined by relating the clinical practice and new research criteria diagnoses to the reference diagnosis. The clinical practice diagnosis at baseline is reflected by a consensus procedure among experts using clinical information only (no biomarkers). The diagnosis based on the new research criteria is reflected by decision rules that combine clinical and biomarker information. The reference diagnosis is determined by a consensus procedure among experts based on clinical information on the course of symptoms over a two-year time period.A decision analytic model is built combining available evidence from different resources among which (accuracy) results from the study, literature and expert opinion to assess long-term cost-effectiveness of the emerging biomarkers. DISCUSSION: Several other multi-centre trials study the relative value of new biomarkers for early evaluation of AD and related disorders. The uniqueness of this study is the assessment of resource utilization and quality of life to enable an economic evaluation. The study results are generalizable to a population of patients who are referred to a memory clinic due to their memory problems.Trial registrationNCT01450891.