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678 result(s) for "Raymond, Philippe"
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موسوعة تاريخ أوربا العام
يتناول كتاب (موسوعة تاريخ أوربا العام) والذي قام بتأليفه (بيار غريمال) في (ثلاث مجلدات) صفحة من القطع المتوسط موضوع (تاريخ أوروبا) مستعرضا أبرز المحتويات التالية : الفصل الأول : مميزات الإطار الجغرافي العام وعوائقه، الفصل الثاني : نماذج من المساحات الإقليمية المميزة، الفصل الثالث : محاور الاستيطان الكثيف للمساحات ومراكز العواصم، الفصل الرابع : أوروبا الحبرية أو عصر المسيحية منتصف القرن الثاني عشر.
Detection of Foodborne Viruses in Dates Using ISO 15216 Methodology
Foodborne viruses such as human norovirus (HuNoV) and hepatitis A virus (HAV) are the major causes of foodborne illnesses worldwide. These viruses have a low infectious dose and are persistent in the environment and food for weeks. Ready-to-eat (RTE) low moisture foods (LMFs) undergo minimal pathogen reduction processes. In recent years, multiple foodborne HAV outbreaks involving hundreds of individuals were associated with the consumption of dates, indicating that they could be important vehicles for foodborne infection. There is no standard method for the extraction and detection of foodborne viruses from dates, but herein we have compared the efficiency of three different protocols based on the ISO 15216 method in the extraction of murine norovirus (MNV) from whole Medjool dates and successfully employed the best performing method in the extraction of HAV, HuNoV GI, and GII and determined the limit of detection (LOD95) of 61, 148, and 184 genomic equivalent (gEq) per 25 g, respectively. Finally, we tested the adopted method on various varieties of dates including pitted ones and reported the detection of HuNoV GI and GII from four naturally contaminated date varieties. This ISO 15216 protocol could be employed for surveillance purposes and outbreak management related to dates.
Evaluation of Porcine Gastric Mucin-Based Method for Extraction of Noroviruses from Seaweed Salad
Human noroviruses (HuNov) are the major cause of foodborne illness globally. Several HuNoV outbreaks have been linked to contaminated ready-to-eat seaweed products. Standard protocols such as the ISO 15216 show limited efficiency in extracting foodborne viruses from seaweed products. Therefore, we evaluated the efficiency of an extraction protocol based on porcine gastric mucin conjugated magnetic beads (PGM-MBs) to recover HuNoVs from Wakame seaweed salad. Compared to other HuNoV extraction methods, the PGM-MB method was more efficient. We then aimed to further improve this protocol by modifying several factors such as the buffers, pH, bead concentration, centrifugation and incubation time. The optimized PGM-MB method yielded 19 ± 3% and 17 ± 4% recovery, for HuNoV GI and GII, respectively. The limit of detection (LOD95) for Wakame seaweed salad was 131 and 56 genomic equivalents per 25 g for HuNoV GI and GII. Although some variability in recovery efficiency was observed between the PGM sources, the optimized PGM-MB protocol effectively extracts HuNoVs from Wakame seaweed salads of various brands and other commodities such as dates, green onions, and salted seaweed. These results support the implementation of the optimized PGM-MB method as a viable alternative for HuNoV surveillance in complex food matrices.
Both Plant and Bacterial Nitrate Reductases Contribute to Nitric Oxide Production in Medicago truncatula Nitrogen-Fixing Nodules
Nitric oxide (NO) is a signaling and defense molecule of major importance in living organisms. In the model legume Medicago truncatula, NO production has been detected in the nitrogen fixation zone of the nodule, but the systems responsible for its synthesis are yet unknown and its role in symbiosis is far from being elucidated. In this work, using pharmacological and genetic approaches, we explored the enzymatic source of NO production in M. truncatula-Sinorhizobium meliloti nodules under normoxic and hypoxic conditions. When transferred from normoxia to hypoxia, nodule NO production was rapidly increased, indicating that NO production capacity is present in functioning nodules and may be promptly up-regulated in response to decreased oxygen availability. Contrary to roots and leaves, nodule NO production was stimulated by nitrate and nitrite and inhibited by tungstate, a nitrate reductase inhibitor. Nodules obtained with either plant nitrate reductase RNA interference double knockdown (MtNR1/2) or bacterial nitrate reductase-deficient (napA) and nitrite reductase-deficient (nirK) mutants, or both, exhibited reduced nitrate or nitrite reductase activities and NO production levels. Moreover, NO production in nodules was found to be inhibited by electron transfer chain inhibitors, and nodule energy state (ATP-ADP ratio) was significantly reduced when nodules were incubated in the presence of tungstate. Our data indicate that both plant and bacterial nitrate reductase and electron transfer chains are involved in NO synthesis. We propose the existence of a nitrate-NO respiration process in nodules that could play a role in the maintenance of the energy status required for nitrogen fixation under oxygen-limiting conditions.
Impact of Capsid and Genomic Integrity Tests on Norovirus Extraction Recovery Rates
Human norovirus (HuNoV) is the leading pathogen responsible for food-borne illnesses. However, both infectious and non-infectious HuNoV can be detected by RT-qPCR. This study evaluated the efficiency of different capsid integrity treatments coupled with RT-qPCR or a long-range viral RNA (long RT-qPCR) detection to reduce the recovery rates of heat inactivated noroviruses and fragmented RNA. The three capsid treatments evaluated (RNase, the intercalating agent PMAxx and PtCl4) reduced the recovery of heat inactivated HuNoV and murine norovirus (MNV) spiked on lettuce, when combined with the ISO 15216-1:2017 extraction protocols. However, PtCl4 also reduced non-heat-treated noroviruses recovery as estimated by RT-qPCR. The PMAxx and RNase treatments had a similar effect on MNV only. The most efficient approaches, the RNase and PMAxx treatments, reduced the heat-inactivated HuNoV recovery rates estimated using RT-qPCR by 2 and >3 log, respectively. The long RT-qPCR detection approach also reduced the recovery rates of heat inactivated HuNoV and MNV by 1.0 and 0.5 log, respectively. Since the long-range viral RNA amplification could be applied to verify or confirm RT-qPCR results, it also provides some advantages by reducing the risk of false positive HuNoV results.
Impact of Nanoparticle-Based TiO2 Surfaces on Norovirus Capsids and Genome Integrity
Human noroviruses (HuNoVs) are among the main causes of acute gastroenteritis worldwide. HuNoVs can survive for several days up to weeks at room temperature in the environment, on food, and on food handling and processing surfaces. As a result, this could lead to viral spread through the ingestion of food in contact with contaminated surfaces. The development of stable surface materials with antiviral activity might be useful to reduce viral outbreaks. Metal-based compounds, including photoactivated titanium nanoparticles (TiO2 NPs), are known for their antiviral activity. In this study, we tested the impact of 2000 µg/mL TiO2 NPs, with or without UV activation, on HuNoV GII and murine norovirus. Their recovery rates were reduced by 99.6%. We also evaluated a new TiO2 NP-coating process on a polystyrene surface. This process provided a homogenous coated surface with TiO2 NPs ranging between 5 nm and 15 nm. Without photoactivation, this TiO2 NP-coated polystyrene surface reduced the recovery rates of intact HuNoV GII by more than 94%. When a capsid integrity treatment with PtCl4 or a longer reverse transcription polymerase chain detection approach was used to evaluate virus integrity following contact with the TiO2 NP-coated polystyrene, the HuNoV GII recovery yield reduction varied between 97 and 100%. These results support the hypothesis that TiO2 NP-coated surfaces have the potential to prevent viral transmission associated with contaminated food surfaces.
A New Substrate Cycle in Plants. Evidence for a High Glucose-Phosphate-to-Glucose Turnover from in Vivo Steady-State and Pulse-Labeling Experiments with 13CGlucose and 14CGlucose
Substrate (futile) cycling involving carbohydrate turnover has been widely reported in plant tissues, although its extent, mechanisms, and functions are not well known. In this study, two complementary approaches, short and steady-state labeling experiments, were used to analyze glucose metabolism in maize (Zea mays) root tips. Unidirectional rates of synthesis for storage compounds (starch, Suc, and cell wall polysaccharides) were determined by short labeling experiments using [U-14C]glucose and compared with net synthesis fluxes to determine the rate of glucose production from these storage compounds. Steady-state labeling with [1-13C]glucose and [U-13C]glucose showed that the redistribution of label between carbon C-1 and C-6 in glucose is close to that in cytosolic hexose-P. These results indicate a high resynthesis flux of glucose from hexose-P that is not accounted for by glucose recycling from storage compounds, thus suggesting the occurrence of a direct glucose-P-to-glucose conversion. An enzyme assay confirmed the presence of substantial glucose-6-phosphatase activity in maize root tips. This new glucose-P-to-glucose cycle was shown to consume around 40% of the ATP generated in the cell, whereas Suc cycling consumes at most 3% to 6% of the ATP produced. The rate of glucose-P cycling differs by a factor of 3 between a maize W22 line and the hybrid maize cv Dea, and is significantly decreased by a carbohydrate starvation pretreatment.
Molecular and Biochemical Characterization of the Involvement of Cyclin-Dependent Kinase A during the Early Development of Tomato Fruit
Abstract Following fruit set, the early development of tomato (Lycopersicon esculentum Mill.) fruit comprises two distinct phases: a cell division phase and a consecutive phase of cell expansion until the onset of ripening. In this study, we analyzed cytological and molecular changes characterizing these early phases of tomato fruit development. First we investigated the spatial and temporal regulation of the mitotic activity during fruit development. The DNA content of isolated nuclei from the different fruit tissues was determined by flow cytometry analysis. The results confirm the data of mitotic activity measurements and show that cell differentiation, leading to expanded cells, is characterized by endoreduplication. Second, we isolated two cDNAs, named Lyces;CDKA1 (accession no. Y17225) and Lyces;CDKA2 (accession no. Y17226), encoding tomato homologs of the cyclin-dependent kinase (CDK) p34cdc2. Tomato CDKA gene expression was followed at both the transcriptional and translational levels during fruit development. The transcripts for Lyces;CDKA1 and Lyces;CDKA2 and the corresponding CDKA proteins are predominantly accumulated during the phase of cell division between anthesis and 5 d post anthesis (DPA). In whole fruits, the maximum CDK activity was obtained between 5 and 10 DPA. The determination of the kinase activity using protein extracts from the different fruit tissues was in agreement with mitotic activity analysis. It showed the particular disappearance of the activity in the gel tissue as early as 15 DPA. The overall data of CDK activity measurements suggest a strong post-translational regulation of CDK at the temporal and spatial levels during early tomato fruit development.
Changes in the expression and the enzymic properties of the 20S proteasome in sugar-starved maize roots. Evidence for an in vivo oxidation of the proteasome
The 20S proteasome (multicatalytic proteinase) was purified from maize (Zea mays L. cv DEA 1992) roots through a five-step procedure. After biochemical characterization, it was shown to be similar to most eukaryotic proteasomes. We investigated the involvement of the 20S proteasome in the response to carbon starvation in excised maize root tips. Using polyclonal antibodies, we showed that the amount of proteasome increased in 24-h-carbon-starved root tips compared with freshly excised tips, whereas the mRNA levels of α3 and β6 subunits of 20S proteasome decreased. Moreover, in carbon-starved tissues, chymotrypsin-like and caseinolytic activities of the 20S proteasome were found to increase, whereas trypsin-like activities decreased. The measurement of specific activities and kinetic parameters of 20S proteasome purified from 24-h-starved root tips suggested that it was subjected to posttranslational modifications. Using dinitrophenylhydrazine, a carbonyl-specific reagent, we observed an increase in carbonyl residues in 20S proteasome purified from starved root tips. This means that 20S proteasome was oxidized during starvation treatment. Moreover, an in vitro mild oxidative treatment of 20S proteasome from non-starved material resulted in the activation of chymotrypsin-like, peptidyl-glutamyl-peptide hydrolase and caseinolytic-specific activities and in the inhibition of trypsin-like specific activities, similar to that observed for proteasome from starved root tips. Our results provide the first evidence, to our knowledge, for an in vivo carbonylation of the 20S proteasome. They suggest that sugar deprivation induces an oxidative stress, and that oxidized 20S proteasome could be associated to the degradation of oxidatively damaged proteins in carbon starvation situations.