Search results for: phosphatases

Authors: Jae-Hyung Jin, Kyung-Tae Lee, Yee-Seul So, Eunji Jeong, Yeonseon Lee, Dongpil Lee, Dong-Gi Lee, Yong-Sun Bahn

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Cryptococcus neoformans causes cryptococcal meningoencephalitis mainly in immunocompromised patients as well as immunocompetent people. But therapeutic options are limited to treat cryptococcosis. Some signaling pathways including cyclic AMP pathway, MAPK pathway, and calcineurin pathway play a central role in the regulation of the growth, differentiation, and virulence of C. neoformans. To understand signaling networks regulating the virulence of C. neoformans, we selected the 114 putative phosphatase genes, one of the major components of signaling networks, in the genome of C. neoformans. We identified putative phosphatases based on annotation in C. neoformans var. grubii genome database provided by the Broad Institute and National Center for Biotechnology Information (NCBI) and performed a BLAST search of phosphatases of Saccharomyces cerevisiae, Aspergillus nidulans, Candida albicans and Fusarium graminearum to Cryptococcus neoformans. We classified putative phosphatases into 14 groups based on InterPro phosphatase domain annotation. Here, we constructed 170 signature-tagged gene-deletion strains through homologous recombination methods for 91 putative phosphatases. We examined their phenotypic traits under 30 different in vitro conditions, including growth, differentiation, stress response, antifungal resistance and virulence-factor production.

Keywords: human fungal pathogen, phosphatase, deletion library, functional genomics

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Authors: Reem Al-Shidhani, Isabelle S. R. Storer, Michael J. Bromley, Lydia Tabernero

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Invasive fungal diseases are an increasing global health concern that contributes to the high mortality rates in immunocompromised patients. The rising of antifungal resistance severely lowers the efficacy of the limited antifungal agents available. New antifungal drugs that target new mechanisms are necessary to tackle the current shortfalls. Amongst post- modifications, phosphorylation is a predominant and an outstanding protein alteration in all eukaryotes. In fungi, protein phosphorylation plays a vital role in many signal transduction pathways, including cell cycle, cell growth, metabolism, transcription, differentiation, proliferation, and virulence. The investigation of Aspergillus fumigatus phosphatases revealed seven genes essential for viability. Inhibiting one of these phosphatases is a new interesting route to develop novel antifungal drugs. In this study, we carried out an early drug discovery process targeting oneessential phosphatase, GlcA. Here, we report the identification of new GlcA inhibitors that show antifungal activity. These important finding open a new avenue to the development of novel antifungals to expand the current narrow arsenal of clinical candidates.

Keywords: invasive fungal diseases, phosphatases, GlcA, competitive inhibitors

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Authors: B. Makoudi, R. Ghanimi, A. Bargaz, M. Mouradi, M. Farissi, A. Kabbaj, J. J. Drevon, C. Ghoulam

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Legume species are able to establish a nitrogen fixing symbiosis with soil rhizobia that allows them, when it operates normally, to ensure their necessary nitrogen nutrition. This biological process needs high phosphorus (P) supply and consequently it is limited under low phosphorus availability. To overcome this constraint, legume-rhizobia symbiosis develops many mechanisms to increase P availability in the rhizosphere and also the efficiency of P fertilizers. The objectives of our research works are to understand the physiological and biochemical mechanisms implemented by legume-rhizobia symbiosis to increase its P use efficiency (PUE) in order to select legume genotypes-rhizobia strains combination more performing for BNF under P deficiency. Our studies were carried out on two grain legume species, common bean (Phaseolus vulgaris) and faba bean (Vicia faba) tested in farmers’ fields and in experimental station fewer than two soil phosphorus levels. Under field conditions, the P deficiency caused a significant decrease of Plant and nodule biomasses in all of the tested varieties with a difference between them. This P limitation increased the contents of available P in the rhizospheric soils that was positively correlated with the increase of phosphatases activities in the nodules and the rhizospheric soil. Some legume genotypes showed a significant increase of their P use efficiency under P deficiency. The P solubilization test showed that some rhizobia strains isolated from Haouz region presented an important capacity to grow on solid and liquid media with tricalcium phosphate as the only P source and their P solubilizing activity was confirmed by the assay of the released P in the liquid medium. Also, this P solubilizing activity was correlated with medium acidification and the excretion of acid phosphatases and phytases in the medium. Thus, we concluded that medium acidification and excretion of phosphatases in the rhizosphere are the prominent reactions for legume-rhizobia symbiosis to improve its P nutrition.

Keywords: legume, phosphorus deficiency, rhizobia, rhizospheric soil

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Authors: B. Makoudi, R. Ghanimi, M. Mouradi, A. Kabbadj, M. Farissi, J. J. Drevon, C. Ghoulam

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This work focuses on the responses of Vicia fabarhizobia symbiosis to phosphorus deficiency and their contribution to tolerate this constraint. The study was carried out on four faba bean varieties, Aguadulce, Alfia, Luz Otono, and Reina Mora submitted to two phosphorus treatments, deficient and sufficient and cultivated under field and greenhouse hydroaeroponic culture. Plants were harvested at flowering stage for growth, nodulation and phosphorus content assessment. Phosphatases in nodules and rhizospheric soil were analyzed. The impact of phosphorus deficiency on yield component was assessed at maturity stage. Under field conditions, phosphorus deficiency affected negatively nodule biomass and nodule phosphorus content with Alfia and Reina Mora showing the highest biomass reduction. The phosphatase activities in nodules and rhizospheric soil were increased under phosphorus deficiency. At maturity stage, under soil low available phosphorus, the pods number and 100 seeds weight were reduced. The genotypic variation was evident for almost all tested parameters.

Keywords: faba bean, phosphorus, rhizobia, yield

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Authors: Neha Sharma, Kanthi K. Kondepudi, Naveen Gupta

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Phytases are phytate specific phosphatases catalyzing the step-wise dephosphorylation of phytate, which acts as an anti-nutritional factor in food due to its strong binding capacity to minerals. In recent years microbial phytases have been explored for improving nutritional quality of food. But the major limitation is acceptability of phytases from these microorganisms. Therefore, efforts are being made to isolate organisms which are generally regarded as safe for human consumption such as Lactic Acid Bacteria (LAB). Phytases from these organisms will have an edge over other phytase sources due to its probiotic attributes. Only few LAB have been reported to give phytase activity that too is generally seen as intracellular. LAB producing extracellular phytase will be more useful as it can degrade phytate more effectively. Moreover, enzyme from such isolate will have application in food processing also. Only few species of Lactobacillus producing extracellular phytase have been reported so far. This study reports the isolation of a probiotic strain of Lactobacillus fermentum spp KA1 which produces extracellular phytase. Conditions for the optimal production of phytase have been optimized and the enzyme production resulted in an approximately 13-fold increase in yield. The phytate degradation potential of extracellular phytase in rice bran has been explored and conditions for optimal degradation were optimized. Under optimal conditions, there was 43.26% release of inorganic phosphate and 6.45% decrease of phytate content.

Keywords: Lactobacillus, phytase, phytate reduction, rice bran

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Authors: Sandeep Kaur, Jenny Vuong, Marcel Julliard, Sean O'Donoghue

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Time-series data are useful for modelling as they can enable model-evaluation. However, when reconstructing models from phosphoproteomic data, often non-exact methods are utilised, as the knowledge regarding the network structure, such as, which kinases and phosphatases lead to the observed phosphorylation state, is incomplete. Thus, such reactions are often hypothesised, which gives rise to uncertainty. Here, we propose a framework, implemented via a web-based tool (as an extension to Minardo), which given time-series phosphoproteomic datasets, can generate κ models. The incompleteness and uncertainty in the generated model and reactions are clearly presented to the user via the visual method. Furthermore, we demonstrate, via a toy EGF signalling model, the use of algorithmic verification to verify κ models. Manually formulated requirements were evaluated with regards to the model, leading to the highlighting of the nodes causing unsatisfiability (i.e. error causing nodes). We aim to integrate such methods into our web-based tool and demonstrate how the identified erroneous nodes can be presented to the user via the visual method. Thus, in this research we present a framework, to enable a user to explore phosphorylation proteomic time-series data in the context of models. The observer can visualise which reactions in the model are highly uncertain, and which nodes cause incorrect simulation outputs. A tool such as this enables an end-user to determine the empirical analysis to perform, to reduce uncertainty in the presented model - thus enabling a better understanding of the underlying system.

Keywords: κ-models, model verification, time-series phosphoproteomic datasets, uncertainty and error visualisation

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Authors: Soumaya Mrabet, Imen Akkari, Amira Atig, Elhem Ben Jazia

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Introduction: Celiac disease (CD) is a chronic autoimmune inflammatory enteropathy caused by gluten. The clinical presentation is very variable. Malabsorption in the MC is responsible for an alteration of the bone metabolism. Our purpose is to study the osteoarticular manifestations related to this condition. Material and methods: It is a retrospective study of 41 cases of CD diagnosed on clinical, immunological, endoscopic and histological arguments, in the Internal Medicine and Gastroenterology Department of Farhat Hached Hospital between September 2005 and January 2016. Results: Osteoarticular manifestations were found in 9 patients (22%) among 41 patients presenting CD. These were 7 women and 2 men with an average age of 35.7 years (25 to 67 years). These manifestations were revelatory of CD in 3 cases. Abdominal pain and diarrhea were present in 6 cases. Inflammatory polyarthralgia of wrists and knees has been reported in 7 patients. Mechanical mono arthralgia was noted in 2 patients. Biological tests revealed microcytic anemia by iron deficiency in 7 cases, hypocalcemia in 5 cases, Hypophosphatemia in 3 cases and elevated alkaline phosphatases in 3 cases. Upper gastrointestinal endoscopy with duodenal biopsy found villous atrophy in all cases. In immunology, Anti-transglutaminase antibodies were positive in all patients, Anti-endomysium in 7 cases. Measurement of bone mineral density (BMD) by biphotonic X-ray absorptiometer with evaluation of the T-score and the Z-score was performed in Twenty patients (48.8%). It was normal in 7 cases (33%) and showed osteopenia in 5 patients (25%) and osteoporosis in 2 patients (10%). All patients were treated with a Gluten-free diet associated with vitamin D and calcium substitution in 5 cases. The evolution was favorable in all cases with reduction of bone pain and normalization of the phosphocalcic balance. Conclusion: The bone impact of CD is frequent but often asymptomatic. Patients with CD should be evaluated by the measurement of bone mineral density and monitored for calcium and vitamin D deficiencies.

Keywords: bone mineral density, celiac disease, osteoarticular manifestations, vitamin D and calcium

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Authors: Kazam Ali, Muhammad Sagheer, Mansoor-Ul- Hasan, Abdul Rashid, Chaudhary Muhammad Shahid Hanif, Fawad Zafar Ahmad Khan, Hafiz Muhammad Aatif

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Natural extracts of two medicinal plants Nicotiana tabacum and Eucalyptus globulus were tested for their toxic and enzyme inhibition effects against three insects species of stored grains Tribolium castaneum, Trogoderma granarium and Sitophilus granarius. Responses of insects varied with exposure periods and dilution levels of acetone extracts of plants. Both plant extracts were lethal to insects but the crude leaf extract of N. tabacum evidenced strong toxic action against three tested insect species. Maximum mortality 36.30% in S. granarius, 25.96% in T. castaneum, and 21.88% in T. granarium were found at 20% dilution level, after 10 days exposure to botanical extract of N. tabacum. The impact of N. tabacum and E. globulus on the activity of esterases; acetylcholinesterase (AChE), α-carboxylesterase (α-CE), β-carboxylesterase (β-CE) and phosphatses; acid phosphatase (AcP), alkaline phosphatase (AlP) of three stored grain insect species were also studied in the survivors of toxicity assay. Whole body homogenates of insects were used for enzyme determination and consumption of high dose rate N. tabacum extract containing diet resulted in maximum 55.33% inhibition of AChE and 26.17% AlP inhibition in T. castaneum, while 44.17% of α-CE and 31.67% inhibition of β-CE activity were noted in S. granarius. Maximum inhibition 23.44% of AcP activity was found in T. granarium exposed to diet treated with the extract of E. globulus. The findings indicate that acetone extracts of N. tabacum and E. globulus are naturally occurring pesticide and facts of the enzyme inhibition relations specify that their effect changes with the insect species.

Keywords: natural extract, medicinal plant, toxic effects, enzyme inhibition, acetone extract

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Authors: Shubhita Mathur, Ritika Kar Bahal, Priyanka Chauhan, Anil K. Tyagi

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Mycobacterium tuberculosis, the causative agent of human tuberculosis, is a major cause of mortality. Bacillus Calmette-Guérin (BCG), the only licensed vaccine available for protection against tuberculosis confers highly variable protection ranging from 0%-80%. Thus, novel vaccine strains need to be evaluated for their potential as a vaccine against tuberculosis. We had previously constructed a triple gene mutant of M. tuberculosis (MtbΔmms), having deletions in genes encoding for phosphatases mptpA, mptpB, and sapM that are involved in host-pathogen interaction. Though vaccination with Mtb∆mms strain induced protection in the lungs of guinea pigs, the mutant strain was not able to control the hematogenous spread of the challenge strain to the spleens. Additionally, inoculation with Mtb∆mms resulted in some pathological damage to the spleens in the early phase of infection. In order to overcome the pathology caused by MtbΔmms in the spleens of guinea pigs and also to control the dissemination of the challenge strain, MtbΔmms was genetically modified by disrupting bioA gene to generate MtbΔmmsb strain. Further, in vivo attenuation of MtbΔmmsb was evaluated, and its protective efficacy was assessed against virulent M. tuberculosis challenge in guinea pigs. Our study demonstrates that Mtb∆mmsb mutant was highly attenuated for growth and virulence in guinea pigs. Vaccination with Mtb∆mmsb mutant generated significant protection in comparison to sham-immunized animals at 4 and 12 weeks post-infection in lungs and spleens of the infected animals. Our findings provide evidence that deletion of genes involved in signal transduction and biotin biosynthesis severely attenuates the pathogen and the single immunization with the auxotroph was able to provide significant protection as compared to sham-immunized animals. The protection imparted by Mtb∆mmsb fell short in comparison to the protection observed in BCG-immunized animals. This study nevertheless indicates the importance of attenuated multiple gene deletion mutants of M. tuberculosis in generating protection against tuberculosis.

Keywords: Mycobacterium tuberculosis, BCG, MtbΔmmsb, bioA, guinea pigs

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Authors: Ayodele A. Otaiku

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Purpose: Nigeria has become the largest importer of rice in Africa and second in the world, 2015. Investigate interactions of organic rice farming on soil quality and health from bio-waste converted to biofertilizer and its environmental impact on rice crop. Methodology: Bio-wastes, poultry waste, organic agriculture wastes, wood ash mixed with microbial inoculant organisms called OBD-Plus microbes (broad spectrum) composted in anaerobic digester to OBD-biofertilizer (2010 - 2012) uses microbes to build humus and other stable carbons. Two field experiments were carried out at Badeggi, Niger state in 2011 and 2012 to evaluate the response of lowland rice production using biofertilizer. The experimental field was laid out in a strip-plot design with five treatments and three replications and at twenty-one day old seedlings of FARO 44 and FARO 52 rice varieties were transplanted. Plots without fertiliser application served as control. Findings: The highest rice grain yield increase of 4.4 t/ha over the control in 2012 against the Nigeria average of lowland rice grain yields of 1.5 t/ha. The utilization of OBD-Biofertilizer can decrease the use of chemical nitrogen fertilizer, prevent the depletion of soil organic matter and reduce environmental pollution. Increasing the floodwater productivity and optimizing the recycling of nutrients cum grazer populations and disease by biocontrols microbes present in the OBD-Biofertilizer. Organic matter in the soil improves by 58% and C/N 15 (2011) and 13.35 (2012). Implications: OBD- Biofertilizer produce plant growth hormones such as indole acetic acid (IAA), glomalin related soil protein and extracellular enzymes as phosphatases that promote soil health and quality. Conclusion: Microorganisms can enhance nutrients use efficiency by increasing root surface area e.g., mycorrhizal, fungi, promoting other beneficial symbioses of the host plant and microbial interactions resulting to increase in soil organic matter. By 2030, climate change is projected to depress cereal production in Africa by 2 to 3 percent. Improved seeds and increased fertilizer use should more than compensate, but this factor will still weigh heavily on efforts to make progress.

Keywords: OBD-plus microbial consortia, OBD-biofertilizer, rice production, soil quality, sustainable agriculture

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