Effect of the Viscosity and Application Time of Different Phosphoric Acids on the Bond Strength and Nanoleakage of Fiber Posts to Root Dentin.

PURPOSE: To evaluate the effect of application time and viscosity of meta-phosphoric acid (MPA) and ortho-phosphoric acid (OPA) on the push-out resin-dentin bond strength (PBS) and nanoleakage (NL) at the adhesive-root dentin interface. METHODS AND MATERIALS: Ninety-six roots of premolars were endodontically prepared and randomly assigned into groups according to the (i) acid (OPA and MPA), (ii) viscosity (gel and liquid), and (iii) application time (7 and 15 seconds). Fiber posts were cemented to the roots, which were then transversally sectioned into serial slices. The slices (cervical, medium, and apical) were subjected to PBS or NL. Analysis of the dentin etching pattern was performed using scanning electron microscopy (SEM). The PBS (MPa) and NL (%) data were subjected to three-way repeated-measures analysis of variance (ANOVA) and Tukey's test (α=0.05). RESULTS: The cross-product interaction was significant for both PBS and NL (p<0.001). No significant difference in the PBS was observed with the use of OPA compared to MPA (p>0.05). Higher PBS values were observed in the groups etched for 15 seconds compared to 7 seconds, regardless of the acid, viscosity, or root third (p<0.05). Lower NL was observed for MPA etching when compared to OPA etching (p<0.05), and these values were not affected by increasing the application time or acid viscosity (p>0.05). A more pronounced etching pattern was observed with OPA than MPA regardless of the acid viscosity. CONCLUSION: The use of 40% meta-phosphoric acid promoted adequate bond strength without increasing dentin demineralization or void spaces in the hybrid layer.

In silico identification of tetraspanins in monopisthocotylean (Platyhelminthes: Monogenea) parasites of fish.

Tetraspanins are a superfamily of transmembrane proteins that in flatworms have structural roles in the development, maturation or stability of the tegument. Several tetraspanins are considered as potential candidates for vaccines or drugs against helminths. Monopisthocotylean monogeneans are ectoparasites of fish that are health hazards for farmed fish. The aim of this study was to identify in silico putative tetraspanins in the genomic datasets of four monopisthocotylean species. The analysis predicted and classified 40 tetraspanins in Rhabdosynochus viridisi, 39 in Scutogyrus longicornis, 22 in Gyrodactylus salaris and 13 in Neobenedenia melleni, belonging to 13 orthologous groups. The high divergence of tetraspanins made it difficult to annotate their function. However, a conserved group was identified in different metazoan taxa. According to this study, metazoan tetraspanins can be divided into 17 monophyletic groups. Of the 114 monogenean tetraspanins, only seven were phylogenetically close to tetraspanins from non-platyhelminth metazoans, which suggests that this group of proteins shows rapid sequence divergence. The similarity of the monopisthocotylean tetraspanins was highest with trematodes, followed by cestodes and then free-living platyhelminths. In total, 27 monopisthocotylean-specific and 34 flatworm-specific tetraspanins were identified. Four monogenean tetraspanins were orthologous to TSP-1, which is a candidate for the development of vaccines and a potential pharmacological target in trematodes and cestodes. Although studies of tetraspanins in parasitic flatworms are scarce, this is an interesting group of proteins for the development of new methods to control monogeneans.

Exhausted T cells in systemic lupus erythematosus patients in long-standing remission.

The mechanisms that drive systemic lupus erythematosus (SLE) patients to achieve remission are unknown; one possible explanation might be T cell exhaustion. The aim of the present study was to measure CD4+ and CD8+ T cell exhaustion in SLE patients in prolonged remission (PR-SLE) and compared them with patients with active SLE (Act-SLE) and healthy subjects. We included 15 PR-SLE patients, 15 Act-SLE and 29 healthy subjects. T cell exhaustion was determined by flow cytometry according to the expression of programmed cell death 1 (PD)-1, T cell immunoglobulin and mucin 3 (Tim-3), natural killer cell receptor (2B4), eomesodermin (EOMES) and T-box transcription factor TBX21 (T-bet) in CD4+ and CD8+ T cells. Dimensionality reduction using the T-distributed stochastic neighbor-embedding algorithm and clustering analysis was used for the identification of relevant populations. Percentages of CD3+ , CD4+ and CD8+ T cells were similar among groups. We identified five subpopulations of CD8+ and seven of CD4+ cells. The CD4+ T-bet+ CD45RO+ cells identified in the unsupervised analysis were significantly increased in PR-SLE versus Act-SLE [median = 0·20, interquartile range (IQR) = 1·74-30·50 versus 1·68, IQR = 0·4-2·83; P < 0·01]. CD4+ EOMES+ cells were also increased in PR-SLE versus Act-SLE (5·24, IQR = 3·38-14·70 versus 1·39, IQR = 0·48-2·87; P < 0·001). CD8+ EOMES+ cells were increased in PR-SLE versus Act-SLE (37·6, IQR = 24·9-53·2 versus 8·13, IQR = 2·33-20·5; P < 0·001). Exhausted and activated T cells presented an increased frequency of PD-1, CD57 and EOMES in SLE patients versus healthy subjects. Some subpopulations of T cells expressing markers associated with exhaustion are increased in patients in remission, supporting T cell exhaustion as a tolerance mechanism in SLE. Exhaustion of specific populations of T cells might represent a potential therapeutic tool that will contribute to the goal of achieving sustained remission in these patients.

Clinical characteristics of systemic lupus erythematosus patients in long-term remission without treatment.

OBJECTIVE: To describe the clinical and serological characteristics of patients with SLE who reached a state of sustained remission for more than 10 years in the absence of treatment. METHODS: From a retrospective cohort of 2121 patients, 44 cases with sustained remission (PtRem) were identified and compared with 88 patients whose course has been chronically active (PtAct).The clinical and serological characteristics were analyzed, as well as the treatment of each group at the beginning of the disease and during its evolution. RESULTS: Older age at disease onset was associated with a tendency to reach a state of prolonged remission. These patients also had a higher frequency of thrombocytopenia at the beginning of the disease 34.1% vs 10.2% (p < 0.001). PtAct had a significantly higher initial SLEDAI compared with cases (10.4 ± 5.6 vs 14.1 ± 5.8; p < 0.001). PtRem had a higher initial frequency of anti-ß2 GP1 IgG antibodies. Also, 25% of these patients were serologically active. We did not find differences in the initial treatment between both groups. The accumulated damage measured by SLICC/ACR damage index at the end of the study was significantly less in the patients who remained in prolonged remission. CONCLUSIONS: Although patients with SLE who achieve prolonged remission have some different characteristics at baseline compared with PtAct, it is not possible to identify a characteristic phenotype for the former. Achieving a state of prolonged remission should always be the goal in patients with SLE. Key Points • SLE patients can reach a very prolonged state of remission, free of treatment, including antimalarials, for at least 10 years. • Venous thromboembolism and thrombocytopenia are commonly present in patients that achieved remission. • The presence of serological markers of activity, even after 10 years in remission, is a risk factor for relapse.

Correction: Clinical characteristics of systemic lupus erythematosus patients in long-term remission without treatment.

The original published version of the above article contained errors in Key Points and Conclusion sections.

Pneumonia in patients with systemic lupus erythematosus: Epidemiology, microbiology and outcomes.

Background and objective Pneumonia remains the main cause of mortality in patients with systemic lupus erythematosus (SLE). The aim of the study was to establish the clinical characteristics, microbiology and risk factors for poor prognosis in patients with SLE and pneumonia. Methods We reviewed medical records of patients with SLE (American College of Rheumatology criteria) and pneumonia who attended the emergency room in a single tertiary care center (January 2010-March 2015). We collected demographics, treatment and disease activity (SLEDAI-2K) data. Severity scales of pneumonia (CURB-65 (acronym for risk factors measured: confusion, urea nitrogen, respiratory rate, blood pressure, 65 years of age and older) and Pneumonia Severity Index (PSI)) were obtained. A negative composite outcome was defined as need for mechanical ventilation, septic shock or death secondary to pneumonia up to 30 days after discharge. We conducted a univariate and multivariable analysis. Results We studied 158 patients (76% women) with 187 episodes of pneumonia. There were no differences in age, SLE duration, SLE activity, treatment or comorbidities between patients with negative composite outcome vs the other group. In 53 episodes, patients presented with a negative composite outcome. Of these, 46 (24.6%) required intubation, 13 (7%) developed shock and 12 (6.4%) died. The most common bacteria isolated was S. aureus, and we observed a high percentage of nonhabitual microorganisms. Fifteen percent of patients who presented with a negative outcome had low values on CURB-65 and PSI scales. Conclusion Patients with SLE and pneumonia have a high risk of complications and present with a high percentage of nonhabitual microorganisms. Severity scales for pneumonia can misclassify as low risk SLE patients with poor prognosis.

A new species of Hemibrycon (Characiformes, Characidae, Stevardiinae) from the upper Magdalena River basin in Colombia.

Hemibrycon iqueima sp. nov., is described from small streams in the Magdalena drainage at the foothills of the western slope of the Eastern Cordillera of the Colombian Andes, Suarez municipality, Tolima Department, Colombia. The new species is distinguished from its congeners in the Magdalena-Cauca River basin by a combination of characters related to snout-anal-fin origin length, head length, dorsal-pectoral fin distance, dorsal-fin-hypural distance, postorbital distance, orbital diameter, snout length, number of total vertebrae, pre-dorsal scales, scale rows between anal-fin origin and lateral line, number of branched rays of the anal fin, maxillary teeth number and number and arrangement of hooks on the branched rays of the pectoral and dorsal fins. In addition, the validity of this species is supported by previous molecular analyses that included specimens of the new species that had been erroneously identified. Phylogenetic relationships between the new species and congeners from Pacific coast basins are discussed.

Controversia en cuanto al uso del icatibant como profilaxis en el angioedema / Controversy to use icatibant for prophylaxis in angioedema

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Mitochondrial dysfunction related to cell damage induced by 3-hydroxykynurenine and 3-hydroxyanthranilic acid: Non-dependent-effect of early reactive oxygen species production.

The kynurenines 3-hydroxyanthranilic acid (3-HANA) and its precursor 3-hydroxykynurenine (3-HK) are metabolites derived from tryptophan degradation. 3-HK, has been related to diverse neurodegenerative diseases including Huntington's, Alzheimer's and Parkinson's diseases that share mitochondrial metabolic dysregulation. Nevertheless, the direct effect of these kynurenines on mitochondrial function has not been investigated despite it could be regulated by their redox properties that are controversial. A body of literature has suggested a ROS mediated cell death induced by 3-HK and 3-HANA. On the other hand, some works have supported that both kynurenines have antioxidant effects. Therefore, the aim of this study was to investigate 3-HK and 3-HANA effects on mitochondrial and cellular function in rat cultured cortical astrocytes (rCCA) and in animals intrastriatally injected with these kynurenines as well as to determinate the ROS role on these effects. First, we evaluated 3-HK and 3-HANA effect on cellular function, ROS production and mitochondrial membrane potential in vivo and in vitro in rCCA. Our results show that both kynurenines decreased MTT reduction in a concentration-dependent manner together with mitochondrial membrane potential. These observations were accompanied with increased cell death in rCCA and in circling behavior and morphological changes of injected animals. Interestingly, we found that ROS production was not increased in both in vitro and in vivo experiments, and accordingly lipid peroxidation (LP) was neither increased in striatal tissue of animals injected with both kynurenines. The lack of effect on these oxidative markers is in agreement with the ·OH and ONOO(-) scavenging capacity of both kynurenines detected by chemical combinatorial assays. Altogether, these data indicate that both kynurenines exert toxic effects through mechanisms that include impairment of cellular energy metabolism which are not related to early ROS production.

Evaluation of specific absorbed fractions from internal photon sources in the ICRP Reference Male Phantom.

In order to estimate the dose caused by internal radiation, it is necessary to know the specific absorbed fraction (SAF) values; through this work these values have been calculated using the Adult Male Reference Computational Phantom (RCP-AM) from the Publication 110 of the International Commission Radiologic Protection and the Monte Carlo transport code MCNPX. These values were calculated for a combination of 980 pairs of source and target organs, for a total of 12 energies. The results were validated and compared with the results reported by other authors: Hadid et al. (RCP-AM), Petoussi-Henss and Zankl (Golem) and the Oak Ridge National Laboratory (ORNL) stylised model reported by Cristy and Eckerman. Mostly, the SAF values calculated with the RCP-AM do not present significant differences in relation to its previous model Golem. When comparing the SAF values of RCP-AM with that of the ORNL stylised model, huge differences were found. These differences can be explained by the shape of the organs and their relative positions, which are more realistic in the voxelised phantoms.

Protein kinase A regulatory subunit isoforms regulate growth and differentiation in Mucor circinelloides: essential role of PKAR4.

The protein kinase A (PKA) signaling pathway plays a role in regulating growth and differentiation in the dimorphic fungus Mucor circinelloides. PKA holoenzyme is comprised of two catalytic (C) and two regulatory (R) subunits. In M. circinelloides, four genes encode the PKAR1, PKAR2, PKAR3, and PKAR4 isoforms of R subunits. We have constructed null mutants and demonstrate that each isoform has a different role in growth and differentiation. The most striking finding is that pkaR4 is an essential gene, because only heterokaryons were obtained in knockout experiments. Heterokaryons with low levels of wild-type nuclei showed an impediment in the emission of the germ tube, suggesting a pivotal role of this gene in germ tube emergence. The remaining null strains showed different alterations in germ tube emergence, sporulation, and volume of the mother cell. The pkaR2 null mutant showed an accelerated germ tube emission and was the only mutant that germinated under anaerobic conditions when glycine was used as a nitrogen source, suggesting that pkaR2 participates in germ tube emergence by repressing it. From the measurement of the mRNA and protein levels of each isoform in the wild-type and knockout strains, it can be concluded that the expression of each subunit has its own mechanism of differential regulation. The PKAR1 and PKAR2 isoforms are posttranslationally modified by ubiquitylation, suggesting another regulation point in the specificity of the signal transduction. The results indicate that each R isoform has a different role in M. circinelloides physiology, controlling the dimorphism and contributing to the specificity of cyclic AMP (cAMP)-PKA pathway.

Synthesis and biological activity of 23-ethylidene-26-hydroxy-22-oxocholestane derivatives from spirostanic sapogenins.

The synthesis and biological evaluation of three new cholestane frameworks of the type: (25R)-3ß,16ß-Diacetoxy-23-ethylidene-26-hydroxy-22-oxocholestane, starting from spirostanic sapogenins of the 25R series, is described. The compounds were obtained by the reductive cleavage of the F ring of 22-oxo-23(1),26-epoxycholestane derivatives using 9-BBN. These modified derivatives exhibit cytotoxic activity against CEM and MCF7 cells and are able to induce apoptosis in them. Its effect on the cell cycle was determined.

Water stress induces up-regulation of DOF1 and MIF1 transcription factors and down-regulation of proteins involved in secondary metabolism in amaranth roots (Amaranthus hypochondriacus L.).

Roots are the primary sites of water stress perception in plants. The aim of this work was to study differential expression of proteins and transcripts in amaranth roots (Amaranthus hypochondriacus L.) when the plants were grown under drought stress. Changes in protein abundance within the roots were examined using two-dimensional electrophoresis and LC/ESI-MS/MS, and the differential expression of transcripts was evaluated with suppression subtractive hybridisation (SSH). Induction of drought stress decreased relative water content in leaves and increased solutes such as proline and total soluble sugars in roots. Differentially expressed proteins such as SOD(Cu-Zn) , heat shock proteins, signalling-related and glycine-rich proteins were identified. Up-regulated transcripts were those related to defence, stress, signalling (Ser, Tyr-kinases and phosphatases) and water transport (aquaporins and nodulins). More noteworthy was identification of the transcription factors DOF1, which has been related to several plant-specific biological processes, and MIF1, whose constitutive expression has been related to root growth reduction and dwarfism. The down-regulated genes/proteins identified were related to cell differentiation (WOX5A) and secondary metabolism (caffeic acid O-methyltransferase, isoflavone reductase-like protein and two different S-adenosylmethionine synthetases). Amaranth root response to drought stress appears to involve a coordinated response of osmolyte accumulation, up-regulation of proteins that control damage from reactive oxygen species, up-regulation of a family of heat shock proteins that stabilise other proteins and up-regulation of transcription factors related to plant growth control.

[Whipple's disease: a case report and review of the literature]. / Enfermedad de Whipple. Presentación de un caso y revisión de la literatura.

Whipple's disease is a systemic infectious condition caused by a gram-positive bacteria belonging to the family of actinomycetes called Tropherima whipplei. It presents most often in males and 13% of patients present with severe diarrhea, fever, adenopathy, weight loss and pigmentation of the skin. Diagnosis is made through duodenal biopsy by which shows foamy macrophages PAS positive. In cases where the histology is negative, we can make the identification of the infectious agent by polymerase-chain reaction (PCR). We describe a 47 years old woman with Whipple's disease whose diagnosis was made through the duodenum and colon biopsies showing infiltration by PAS-positive macrophages.

Altitudinal variation in fish assemblage diversity in streams of the central Andes of Colombia.

This study documents differences in fish assemblages for 32 freshwater streams located between 258 and 2242 m a.s.l. on the eastern slopes of the central range of the Colombian Andes. A total of 2049 fishes belonging to 62 species, 34 genera and 16 families were collected. Species richness declined rapidly with altitude; nearly 90% of the species were recorded between 250 and 1250 m a.s.l. Three of the four physico-chemical variables, of the water, temperature, dissolved oxygen and pH, explained 53.5% of the variation in species richness along the altitudinal gradient, with temperature the most important (37.6%). An analysis of species composition showed that the distinctiveness of the fish fauna increased with elevation, with the greatest turnover observed between 1000 and 1750 m a.s.l. On this altitudinal gradient, turnover was dominated by the loss of species rather than gain, and dominance by just a few species was greater at higher elevations. Turnover was also observed along the altitudinal gradient in the structure of the three functional groups (torrential, pool and pelagic species). The study focused on understanding the pattern of diversity of fish communities inhabiting the Andes in Colombia. Anthropogenic effects on the altitudinal distribution of fish species in the region, however, are largely unknown and would require further investigations.

Comparación de cuatro métodos de medición de la tasa de filtración glomerular con depuración de inulina en individuos sanos y en pacientes con insuficiencia renal / Comparison of Four Methods for Measuring Glomerular Filtration Rate by Inulin Clearance in Healthy Individuals and Patients with Renal Failure

Introducción. La medición de la función renal es importante para el diagnostico y estratificación de las enfermedad renales. Varios métodos han sido empleados para predecir la tasa de filtración glomerular, sin embargo los resultados han sido variables según la población estudiada. En este estudio se compararon 4 métodos de medición de la función renal con la depuración de inulina. Métodos. Se realizó depuración de inulina, tecnecio y creatinina y se calculó la filtración glomerular con las fórmulas de Cockcroft-Gault y Levey en 51 sujetos con función renal estable. El análisis estadístico se realizó mediante el coeficiente de correlación de Pearson y con análisis de concordancia (método de Bland y Altman). Resultados. Se incluyeron 51 sujetos de los cuales 35 (68.6%) se encontraban con algún grado de insuficiencia renal. Los 4 métodos evaluados mostraron una correlación significativa con la depuración de inulina. Sin embargo, todos mostraron considerable falta de concordancia, con límites inferiores que variaban desde 15 hasta 42 ml/min, para las comparaciones de Inulina con tecnecio e inulina con la fórmula de Levey, respectivamente, y límites de concordancia superiores que podían ir desde 20 y hasta 56 ml/min, para las comparaciones de Inulina con tecnecio e Inulina con la fórmula de Levey, respectivamente. Conclusiòn. La medición de la tasa de filtración glomerular efectuada a través de diferentes métodos muestra un amplio rango de variación al compararlos con la depuración de inulina, lo cual debe de considerarse en la práctica clínica diaria al evaluar la función renal (AU)

Background. A proper measurement of renal function is important for diagnosis and stratification of kidney disease. Several methods have been used to predict glomerular filtration rate, however the results have been variable depending on the population studied. We aimed to compare the performances of 4 glomerular filtration rate tests with inulin clearance in patients with chronic renal insufficiency and in healthysubjects. Methods. Inulin clearances performed in 51 individuals with stable renal function were selected. For each of them, we computed 4 estimates: the 24-hour creatinine clearance, technetium (99mTc-DTPA) clearance, Cockcroft-Gault and Levey formulas. Their respective performance was assessed by correlation (Pearson's correlation coefficient) and agreement (Bland and Altman method). Results. Each glomerular filtration rate test closely correlated with inulin clearance. Nevertheless, all GFR tests displayed considerable lack of agreement with lower limits ranging from 15 to 42 ml / min, for comparison with inulin-technetium and inulin with Levey formula, respectively and upper limits of agreement that could range from 20 to 56 ml / min, for comparison with inulin-technetium and Inulin with Levey formula,respectively. Conclusion. The measurement of glomerular filtration rate determined via different methods shows a wide range of variation when compared with inulin clearance, which should be considered in daily clinical practice during the evaluation of renal function (AU)

[Comparison of four methods for measuring glomerular filtration rate by inulin clearance in healthy individuals and patients with renal failure]. / Comparación de cuatro métodos de medición de la tasa de filtración glomerular con depuración de inulina en individuos sanos y en pacientes con insuficiencia renal.

BACKGROUND: A proper measurement of renal function is important for diagnosis and stratification of kidney disease. Several methods have been used to predict glomerular filtration rate, however the results have been variable depending on the population studied. We aimed to compare the performances of 4 glomerular filtration rate tests with inulin clearance in patients with chronic renal insufficiency and in healthy subjects. METHODS: Inulin clearances performed in 51 individuals with stable renal function were selected. For each of them, we computed 4 estimates: the 24-hour creatinine clearance, technetium (99mTc-DTPA) clearance, Cockcroft-Gault and Levey formulas. Their respective performance was assessed by correlation (Pearson's correlation coefficient) and agreement (Bland and Altman method). RESULTS: Each glomerular filtration rate test closely correlated with inulin clearance. Nevertheless, all GFR tests displayed considerable lack of agreement with lower limits ranging from 15 to 42 ml / min, for comparison with inulin-technetium and inulin with Levey formula, respectively and upper limits of agreement that could range from 20 to 56 ml / min, for comparison with inulin-technetium and Inulin with Levey formula,respectively. CONCLUSION: The measurement of glomerular filtration rate determined via different methods shows a wide range of variation when compared with inulin clearance, which should be considered in daily clinical practice during the evaluation of renal function.

Contribution of the saprobic fungi Trametes versicolor and Trichoderma harzianum and the arbuscular mycorrhizal fungi Glomus deserticola and G. claroideum to arsenic tolerance of Eucalyptus globulus.

The presence of high concentrations of arsenic (As) decreased the shoot and root dry weight, chlorophyll and P and Mg content of Eucalyptus globulus colonized with the arbuscular mycorrhizal (AM) fungi Glomus deserticola or G. claroideum, but these parameters were higher than in non-AM plants. As increased the percentage of AM length colonization and succinate dehydrogenase (SDH) activity in the root of E. globulus. Trichoderma harzianum, but not Trametes versicolor, increased the shoot and root dry weight, chlorophyll content, the percentage of AM root length colonization and SDH activity of E. globulus in presence of all As concentrations applied to soil when was inoculated together with G. claroideum. AM fungi increased shoot As and P concentration of E. globulus to higher level than the non-AM inoculated controls. The contribution of the AM and saprobe fungi to the translocation of As from root to shoot of E. globulus is discussed.

Interactions of Trametes versicolor, Coriolopsis rigida and the arbuscular mycorrhizal fungus Glomus deserticola on the copper tolerance of Eucalyptus globulus.

The presence of high levels of Cu in soil decreases the shoot and root dry weights of Eucalyptus globulus. However, higher plant tolerance of Cu has been observed in the presence of the arbuscular mycorrhizal (AM) fungus Glomus deserticola. The hyphal length of G. deserticola was sensitive to low Cu concentrations, and the percentage of AM root colonisation and the metabolic activity of the AM fungus were also decreased by Cu. Therefore, a direct effect of Cu on the development of the AM fungus inside and outside the root cannot be ruled out. E. globulus colonised by G. deserticola had higher metal concentrations in the roots and shoots than do non-mycorrhizal plants; however, the absence of a higher root to shoot metal ratio in the mycorrhizal plants (1.70+/-0.11) indicated that G. deserticola did not play a filtering/sequestering role against Cu. The saprobe fungi Coriolopsis rigida and Trametes versicolor were able to remove Cu ions from the asparagine-glucose growth media. However, plants inoculated with C. rigida and T. versicolor did not accumulate more Cu than non-inoculated controls, and the growth of the plant was not increased in the presence of these fungi. However, C. rigida increased the shoot dry weight, AM root length colonisation, and metabolic mycelial activity of plants colonised with G. deserticola in the presence of Cu; only this saprobe-AM fungus combination increased the tolerance of E. globulus to Cu. Inoculation with G. deserticola and C. rigida increased the E. globulus Cu uptake to levels reached by hyperaccumulative plants.