Calcium Oxalate Crystals in Eucalypt Ectomycorrhizae: Morphochemical Characterization

Ectomycorrhizal fungi are ubiquitous in forest ecosystems, benefitting plants principally by increasing the uptake of water and nutrients such as calcium from the soil. Previous work has demonstrated accumulation of crystallites in eucalypt ectomycorrhizas, but detailed morphological and chemical characterization of these crystals has not been performed. In this work, cross sections of acetic acid-treated and cleared ectomycorrhizal fragments were visualized by polarized light microscopy to evaluate the location of crystals within cortical root cells. Ectomycorrhizal sections were also observed by scanning electron microscopy (SEM) coupled with energy dispersive x-ray (EDS) microprobe analysis. The predominant forms of crystals were crystal sand (granules) and concretions. Calcium, carbon and oxygen were detected by EDS as constituent elements and similar elemental profiles were observed between both crystal morphologies. All analyzed crystalline structures were characterized as calcium oxalate crystals. This is the first report of the stoichiometry and morphology of crystals occurring in eucalypt ectomycorrhizas in tropical soils. The data corroborates the role of ectomycorrhizae in the uptake and accumulation of calcium in the form of calcium oxalate crystals in hybrid eucalypt plants.     

Herpes simplex type 1 activates glycolysis through engagement of the enzyme 6-phosphofructo-1-kinase (PFK-1)

Viruses such as HIV, HCV, Mayaro and HCMV affect cellular metabolic pathways, including glycolysis. Although some studies have suggested that the inhibition of glycolysis affects HSV-1 replication and that HSV-1-infected eyes have increased lactate production, the mechanisms by which HSV-1 induces glycolysis have never been investigated in detail. In this study, we observed an increase in glucose uptake, lactate efflux and ATP content in HSV-1-infected cells. HSV-1 triggered a MOI-dependent increase in the activity of phosphofructokinase-1 (PFK-1), a key rate-limiting enzyme of the glycolytic pathway. After HSV-1 infection, we observed increased PFK-1 expression, which increased PFK-1 total activity, and the phosphorylation of this enzyme at serine residues. HSV-1-induced glycolysis was associated with increased ATP content, and these events were critical for viral replication. In summary, our results suggest that HSV-1 triggers glycolysis through a different mechanism than other herpesviruses, such as HCMV. Thus, this study contributes to a better understanding of HSV-1 pathogenesis and provides insights into novel targets for antiviral therapy. HIGHLIGHTS: ?HSV-1 activates glycolysis by PFK-1 activation. ?In HSV-1-infected cells PFK-1 synthesis is up-regulated and phosphorylated at serine residues. ?PFK-1 knockdown impairs HSV-1 replication. ?HSV-1-mediated glycolysis activation increases ATP content.     

Effect of Gregarina sp. parasitism on the susceptibility of Blattella germanica to some control agents

Gregarines are enteric parasites of invertebrates but little is known about the negative effects of this parasitism on host species. The present study evaluates the influence of the parasitism of Gregarina sp. on the survival of Blattella germanica and methods for elimination of gregarine infection in laboratory rearing systems. Insects were dissected and the infection was detected in 80% of a sample of 50 adults. Diseased cockroaches had swollen abdomens, slower movement at high incidences of the protozoan, and short antennas. Dead cockroaches showed darkened body and putrid smell, indicating septicaemia. Infected insects were more susceptible than healthy cockroaches when treated with Metarhizium anisopliae and triflumuron.     

Metabolic profiling by ultra-performance liquid chromatography-mass spectrometry and parallel factor analysis for the determination of disease biomarkers in Eucalyptus

In this article, we present and discuss an alternative for data analysis of the metabolic profiles of both healthy Eucalyptus globulus and those infected with the Mycosphaerella leaf disease. The crude extracts were analyzed by reversed-phase ultra performance liquid chromatography-mass spectrometry. In order to glean the most useful information from these complex measurements, parallel factor analysis (PARAFAC) was employed for pattern recognition. After PARAFAC modeling, inspection of the scores and loadings graph allowed distinction of the healthy from the infected E. globulus samples and determination of biomarkers related to the biotic stress. The assessment of the monoisotopic masses and the fragmentation patterns allowed the identification of these biomarkers. It is hoped that the proposed method can be used for the diagnosis of diseases in plants, as well as to provide additional insight into the plant’s defense mechanism. Potentially, this may demonstrate the advantages of employing high order chemometric techniques in metabolomic data analysis.     

Retinol-induced elevation of ornithine decarboxylase activity in cultured rat Sertoli cells is attenuated by free radical scavenger and by iron chelator

We investigated retinol effects in ornithine decarboxylase activity in Sertoli cells. We also tested the hypothesis that free radical scavengers and iron chelators may attenuate the effect of retinol. Sertoli cells isolated from 15-day-old Wistar rats were previously cultured for 48 h and then treated with retinol by 24 h with or without mannitol (1 mM) or 1,10 phenanthroline (100 M). We measured ornithine decarboxylase and catalase activities and malondialdehyde concentrations in response to retinol treatment. In response to 7 M retinol treatment ornithine decarboxylase activity increased 30%. Retinol-induced ornithine decarboxylase activity was significantly decreased by addition of free radical scavenger (mannitol) or iron chelator (1,10 phenanthroline). In addition the same effect was observed in catalase increased activity and in malondialdehyde concentrations. These results suggest that retinol treatment induced ornithine decarboxylase and catalase activity and increased malondialdehyde concentration. These effects appear to be mediate by ROS.     

Refolding and characterization of methionine adenosyltransferase from Euglena gracilis

Methionine adenosyltransferase from Euglena gracilis (MATX) is a recently discovered member of the MAT family of proteins that synthesize S-adenosylmethionine. Heterologous overexpression of MATX in Escherichia coli rendered the protein mostly in inclusion bodies under all conditions tested. Therefore, a refolding and purification procedure from these aggregates was developed to characterize the enzyme. Maximal recovery was obtained using inclusion bodies devoid of extraneous proteins by washing under mild urea (2M) and detergent (5%) concentrations. Refolding was achieved in two steps following solubilization in the presence of Mg(2+); chaotrope dilution to <1M and dialysis under reducing conditions. Purified MATX is a homodimer that exhibits Michaelis kinetics with a V(max) of 1.46 μmol/min/mg and K(m) values of approximately 85 and 260 μM for methionine and ATP, respectively. The activity is dependent on Mg(2+) and K(+) ions, but is not stimulated by dimethylsulfoxide. MATX exhibits tripolyphosphatase activity that is stimulated in the presence of S-adenosylmethionine. Far-UV circular dichroism revealed β-sheet and random coil as the main secondary structure elements of the protein. The high level of sequence conservation allowed construction of a structural model that preserved the main features of the MAT family, the major changes involving the N-terminal domain.     

Fungi isolated from olive ecosystems and screening of their potential biotechnological use

This study investigated the fungi diversity of fresh olive (Olea europaea L.) fruits, olive paste (crushed olives) and olive pomace (solid waste) and screened and quantified enzymatic activities with biotechnological applications. Fungi were randomly isolated from olive cultivars from Castilla La Mancha region (Spain). Identification included comparison of their polymerase chain reaction (PCR) amplicons of the ITS1-5.8S-ITS2 ribosomal DNA region, followed by nucleotide sequence analysis. Fourteen different species with DNA sequences of different similarities were identified, belonging to seven different genera (Aspergillus, Penicillium, Rhizomucor, Mucor, Rhizopus, Lichtheimia and Galactomyces). Aspergillus fumigatus, followed by Galactomyces geotrichum, Penicillium commune and Rhizomucor variabilis var. regularior were the most frequent species. Specific enzyme screening was assayed on agar plates, using cellobiose, carboxymethylcellulose (CMC), polygalacturonic acid and CaCl(2)/Tween 80 as substrates for β-glucosidase, carboxymethylcellulase (CMCase), polygalacturonase and lipase, respectively. Species exhibiting the best activities were: Aspergillus fumigatus (for β-glucosidase, CMCase and lipase); Rhizopus oryzae (for β-glucosidase and lipase); Rhizomucor variabilis (for β-glucosidase, CMCase and polygalacturonase); Mucor fragilis (β-glucosidase, CMCase and lipase); Galactomyces geotrichum (for β-glucosidase, polygalacturonase and lipase) and Penicillium commune and Penicillium crustosum (for lipase). The species that had shown the best enzymatic activities were grown on hemicellulose, cellulose and pectin and some activities were quantified (xylanase, cellulase, β-glucosidase and pectinase). An isolate of A. fumigatus and one of A. niger showed the best cellulase and xylanase activities, while no species presented good pectinase and β-glucosidase activities. The selected species with potential enzymatic activities could be used for future applications of industrial interest.     

Beauveria Bassiana Pathogenicity to the Citrus Rust Mite Phyllocoptruta Oleivora

The pathogenicity of Beauveria bassiana to one of the major pests of citrus crops, Phyllocoptruta oleivora, was assessed by inoculating mites with different concentrations of conidia (1×10⁶, 5×10⁶, 1×10⁷, 5×10⁷ and 1×10⁸). Treated mites were kept at controlled conditions (25 ± 0.5°C, 12 h photoperiod and 98% relative humidity) and mite survivorship was evaluated daily. Mortality was found to increase in time and was dependent on the conidia concentration, with values ranging from 24 to 91% for the lowest and highest conidia concentration, respectively. The calculated LC₅₀ on the fifth day was 4.23×10⁶ conidia/ml. Mean lethal time was 3.98, 9.79, 3.09 and 2.74 days for 5×10⁶, 1×10⁷, 5×10⁷ and 1×10⁸ conidia/ml, respectively. Conidia were found to adhere all over the mite body surface, especially at the anal region, where vegetative mycelium was found entering the mite body. We noticed the formation of small crystals inside the mite’s body that were produced during colonization of the body cavity by the fungus. This is the first report of B. bassiana pathogenicity for this species.     

Augmented cell survival in eutopic endometrium from women with endometriosis: Expression of c-myc,TGF-beta1 and bax genes

Background  

Endometriosis is a common gynaecological disorder characterized by the presence of endometrial tissue outside of the uterus. The fragments in normal menstruation are composed of necrotic and living cells, which do not survive in ectopic locations because of programmed cell death. The aim of this study was to evaluate if the balance between cell proliferation and apoptosis is changed in eutopic endometrium from women with endometriosis throughout the menstrual cycle by studying bax (pro-apoptotic), c-myc (regulator of cell cycle) and TGF-beta1 (involved in cell differentiation) genes.