Manganese accumulation in rice: implications for photosynthetic functioning

In order to gain fundamental insights into the nature of the adaptation to Mn excess, the characterisation of the photosynthetic apparatus in Mn-treated rice was carried out in 21-day-old plants. We found 17- and 11-fold increases in Mn in the leaf tissues and in thylakoid, respectively, when the plants were grown hydroponically in nutrient solutions with Mn concentrations between 0.125 and 32 mg l(-1) (2.3 and 582.5 microM). Net photosynthesis and the photosynthetic capacity decreased after the 0.5 and 2 mg l(-1) (9.1 and 36.4 microM) Mn treatment, respectively. The stomatal conductance displayed a similar trend to that of photosynthetic capacity. The levels of basal chlorophyll fluorescence and the ratio between variable and maximum chlorophyll fluorescence did not vary significantly among treatments, but the photochemical quenching and the quantum yield of non-cyclic electron transport increased until the 2 mg l(-1) (36.4 microM) Mn treatment. The lipid matrix of thylakoids revealed a global increase in the proportions of phospholipids, relative to galactolipids. This pattern was coupled with diminishing levels of monogalactosyldiacylglycerol. The relative ratio between total carotenoids and total chlorophylls decreased until the last Mn treatment, yet the levels of carotenes, zeaxanthin, and violaxanthin plus antheraxanthin displayed different patterns. It was further found that the de-epoxidation state involving the components of the xanthophylls cycle increased until the 8 mg l(-1) (145.6 microM) Mn treatment. The levels of the photosynthetic electron carriers displayed different patterns, with plastocyanin and the high and low forms of cytochrome b559 remaining steady, whereas cytochromes b563 and f increased until the 8 mg l(-1) (145.6 microM) Mn treatment and the quinone pool increased until the highest Mn treatment. It was concluded that Mn-mediated inhibition of rice photosynthesis barely implicates stomatal conductance, as well as the distribution of energy within the photosystems. In this context, alterations to the relative proportions of the different acyl lipids and isoprenoids, as well as to the accumulations of the photosynthetic electron carriers, seem to play a major role.     

Evaluation of a Western blot test,Helico blot 2.1, in the diagnosis of Helicobacter pylori infection in a pediatric population

Background. Noninvasive diagnostic tests are useful as screening tools for Helicobacter pylori infection in pediatric populations. The aim of this study was to evaluate performance of the immunoblot assay, Helico Blot 2.1, for the diagnosis of H. pylori infection in symptomatic children. Materials and Methods. Immunoblot assay was used for detection of IgG antibodies to specific H. pylori proteins and to a recombinant H. pylori antigen, CIM marker. The study was performed on sera collected from 134 symptomatic, untreated children (mean age, 9.1 ± 3.2 years; range, 1–14 years). H. pylori infection status was determined by culture, histology and rapid urease test. Results. Immunoblot assay yielded a positive result in 71 of the 72 infected patients (sensitivity 98.6%) and in eight of the 62 noninfected ones (specificity 87.1%). The predictive values for a positive and a negative result were 89.9% and 98.2%, respectively. The performance of the CIM band alone, as a marker for H. pylori infection status, was also evaluated. This band was present on the blot of 71 infected patients and on four of the 62 H. pylori‐negative patients. The sensitivity, specificity, PPV and NPV of the CIM antigen were 98.6%, 93.5%, 94.7% and 98.3%, respectively. Conclusions. The immunoblot assay Helico Blot 2.1 is a suitable noninvasive test for the serodiagnosis of H. pylori infection in children. The good level of performance demonstrated by the novel recombinant antigen CIM suggests it may be a useful contribution to the qualitative and quantitative performance of the Helico Blot 2.1 in pediatric populations.     

Characterization of azo reduction activity in a novel ascomycete yeast strain

Several model azo dyes are reductively cleaved by growing cultures of an ascomycete yeast species, Issatchenkia occidentalis. In liquid media containing 0.2 mM dye and 2% glucose in a mineral salts base, more than 80% of the dyes are removed in 15 h, essentially under microaerophilic conditions. Under anoxic conditions, decolorization does not occur, even in the presence of pregrown cells. Kinetic assays of azo reduction activities in quasi-resting cells demonstrated the following: (i) while the optimum pH depends on dye structure, the optimum pH range was observed in the acidic range; (ii) the maximum decolorizing activity occurs in the late exponential phase; and (iii) the temperature profile approaches the typical bell-shaped curve. These results indirectly suggest the involvement of an enzyme activity in azo dye reduction. The decolorizing activity of I. occidentalis is still observed, although at a lower level, when the cells switch to aerobic respiration at the expense of ethanol after glucose exhaustion in the culture medium. Decolorization ceased when all the ethanol was consumed; this observation, along with other lines of evidence, suggests that azo dye reduction depends on cell growth. Anthraquinone-2-sulfonate, a redox mediator, enhances the reduction rates of the N,N-dimethylaniline-based dyes and reduces those of the 2-naphthol-based dyes, an effect which seems to be compatible with a thermodynamic factor. The dye reduction products were tested as carbon and nitrogen sources. 1-Amino-2-naphthol was used as a carbon and nitrogen source, and N,N-dimethyl-p-phenylenediamine was used only as a nitrogen source. Sulfanilic and metanilic acids did not support growth either as a carbon or nitrogen source.     

Evaluation of rapid test Assure Helicobacter pylori for diagnosis of H. pylori in pediatric population

Non-invasive tests are needed to assess Helicobacter pylori infection, especially to screen a pediatric population. Assure H. pylori Rapid Test (Genelabs Diagnostics, Singapore) is an immunochromatographic assay device intended for the rapid detection of antibodies to H. pylori in human serum, plasma or whole blood. The aim of this study was to evaluate the performance of the rapid test, Assure H. pylori, in the diagnosis of H. pylori infection in children, using a Portuguese pediatric population. The study group included 130 children with age ranging from 1 to 14 years old (average age 9.2+/-3.1 years). According to the gold standard, 70 of the 130 patients studied were H. pylori positive and 60 were H. pylori negative. Using Assure H. pylori Rapid Test (Genelabs Diagnostics, Singapore), 53 sera had a positive result after 15 min (resulting in 17 false negatives) and 57 sera a negative result (resulting in 3 false positives). The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the test were 75.7%, 95.0%, 94.6% and 77.0% respectively. When a longer read time of 45 min is considered, the rapid test revealed a good performance (sensitivity 98.6% and specificity 95%) in the evaluation of the H. pylori infection in a pediatric population. In conclusion, the test showed a good performance, suggesting its applicability as a screening method for the H. pylori infection.     

Interspecific Associations between Cycloneda sanguinea and Two Aphid Species (Aphis gossypii and Hyadaphis foeniculi) in Sole-Crop and Fennel-Cotton Intercropping Systems

Aphids cause significant damage to crop plants. Studies regarding predator-prey relationships in fennel (Foeniculum vulgare Mill.) and cotton (Gossypium hirsutum L.) crops are important for understanding essential ecological interactions in the context of intercropping and for establishing pest management programs for aphids. This study evaluated the association among Hyadaphis foeniculi (Passerini) (Hemiptera: Aphididae), Aphis gossypii Glover (Hemiptera: Aphididae) and Cycloneda sanguinea (L.) (Coleoptera: Coccinellidae) in cotton with coloured fibres, fennel and cotton intercropped with fennel. Association analysis was used to investigate whether the presence or absence of prey and predator species can indicate possible interactions between aphids and ladybugs. Significant associations among both apterous and alate H. foeniculi and C. sanguinea were observed in both the fennel and fennel-cotton intercropping systems. The similarity analysis showed that the presence of aphids and ladybugs in the same system is significantly dependent on the type of crop. A substantial amount of evidence indicates that the presence of the ladybug C. sanguinea, is associated with apterous or alate A. gossypii and H. foeniculi in fennel-cotton intercropping system. We recommend that future research vising integrated aphid management taking into account these associations for take decisions.     

Mitogen activated protein kinases SakAHOG1 and MpkC collaborate for Aspergillus fumigatus virulence

Here, we investigated which stress responses were influenced by the MpkC and SakA mitogen‐activated protein kinases of the high‐osmolarity glycerol (HOG) pathway in the fungal pathogen Aspergillus fumigatus. The ΔsakA and the double ΔmpkC ΔsakA mutants were more sensitive to osmotic and oxidative stresses, and to cell wall damaging agents. Both MpkC::GFP and SakA::GFP translocated to the nucleus upon osmotic stress and cell wall damage, with SakA::GFP showing a quicker response. The phosphorylation state of MpkA was determined post exposure to high concentrations of congo red and Sorbitol. In the wild‐type strain, MpkA phosphorylation levels progressively increased in both treatments. In contrast, the ΔsakA mutant had reduced MpkA phosphorylation, and surprisingly, the double ΔmpkC ΔsakA had no detectable MpkA phosphorylation. A. fumigatus ΔsakA and ΔmpkC were virulent in mouse survival experiments, but they had a 40% reduction in fungal burden. In contrast, the ΔmpkC ΔsakA double mutant showed highly attenuated virulence, with approximately 50% mice surviving and a 75% reduction in fungal burden. We propose that both cell wall integrity (CWI) and HOG pathways collaborate, and that MpkC could act by modulating SakA activity upon exposure to several types of stresses and during CW biosynthesis.     

High osmolarity glycerol response PtcB phosphatase is important for Aspergillus fumigatus virulence

Aspergillus fumigatus is a fungal pathogen that is capable of adapting to different host niches and to avoid host defenses. An enhanced understanding of how, and which, A. fumigatus signal transduction pathways are engaged in the regulation of these processes is essential for the development of improved disease control strategies. Protein phosphatases are central to numerous signal transduction pathways. To comprehend the functions of protein phosphatases in A. fumigatus, 32 phosphatase catalytic subunit encoding genes were identified. We have recognized PtcB as one of the phosphatases involved in the high osmolarity glycerol response (HOG) pathway. The ΔptcB mutant has both increased phosphorylation of the p38 MAPK (SakA) and expression of osmo‐dependent genes. The ΔptcB strain was more sensitive to cell wall damaging agents, had increased chitin and β‐1,3‐glucan, and impaired biofilm formation. The ΔptcB strain was avirulent in a murine model of invasive pulmonary aspergillosis. These results stress the importance of the HOG pathway in the regulation of pathogenicity determinants and virulence in A. fumigatus.