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BACKGROUND: Treatment for haemophilia A has expanded from plasma-derived factor VIII (pdFVIII) and standard half-life (SHL) recombinant FVIII (rFVIII) to extended half-life (EHL) rFVIII and non-factor products that mimic FVIII activity. OBJECTIVE: To determine amounts of clotting factor concentrates (CFCs) and emicizumab dispensed and associated healthcare expenditures in Japanese patients with haemophilia A. METHODS: This retrospective, non-interventional, observational study analysed data from 2016 to 2020 from a large-scale, hospital-based administrative database. Patients had haemophilia A without inhibitors and ≥ 2 prescriptions of the same CFC or emicizumab. RESULTS: In total, 974 patients with haemophilia A (median age, 30.0 years; median follow-up, 3.7 years) were included. Outpatient use of EHL rFVIII and emicizumab increased, although pdFVIII/SHL rFVIII were still used over the study. Median annual total healthcare expenditures/patient increased from ¥9,200,230 in 2016 to ¥19,748,221 in 2020. Overall, the median annual drug expenditure/patient increased from ¥8,723,120 in 2016 to ¥18,051,689 in 2020. Drug expenditure was highest with emicizumab, with an increase in median annual expenditure/patient from 2018 (n = 4, ¥26,030,206) to 2020 (n = 107, ¥45,430,408). Overall, 233 patients (23.9%) switched from an SHL to an EHL product. Although amounts of FVIII prescribed increased in the 3 months after switching, overall, there was no noticeable difference before and after switching. Median total healthcare and FVIII product expenditures increased following switching. CONCLUSIONS: Prescribing of EHL products increased over the study and healthcare expenditures increased for patients who switched from SHL to EHL rFVIII products.
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Wood formation involves consecutive developmental steps, including cell division of vascular cambium, xylem cell expansion, secondary cell wall (SCW) deposition, and programmed cell death. In this study, we identified PagMYB31 as a coordinator regulating these processes in Populus alba × Populus glandulosa and built a PagMYB31-mediated transcriptional regulatory network. PagMYB31 mutation caused fewer layers of cambial cells, larger fusiform initials, ray initials, vessels, fiber and ray cells, and enhanced xylem cell SCW thickening, showing that PagMYB31 positively regulates cambial cell proliferation and negatively regulates xylem cell expansion and SCW biosynthesis. PagMYB31 repressed xylem cell expansion and SCW thickening through directly inhibiting wall-modifying enzyme genes and the transcription factor genes that activate the whole SCW biosynthetic program, respectively. In cambium, PagMYB31 could promote cambial activity through TRACHEARY ELEMENT DIFFERENTIATION INHIBITORY FACTOR (TDIF)/PHLOEM INTERCALATED WITH XYLEM (PXY) signaling by directly regulating CLAVATA3/ESR-RELATED (CLE) genes, and it could also directly activate WUSCHEL HOMEOBOX RELATED4 (PagWOX4), forming a feedforward regulation. We also observed that PagMYB31 could either promote cell proliferation through the MYB31-MYB72-WOX4 module or inhibit cambial activity through the MYB31-MYB72-VASCULAR CAMBIUM-RELATED MADS2 (VCM2)/PIN-FORMED5 (PIN5) modules, suggesting its role in maintaining the homeostasis of vascular cambium. PagMYB31 could be a potential target to manipulate different developmental stages of wood formation.
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Câmbio , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Populus , Fatores de Transcrição , Xilema , Populus/genética , Populus/crescimento & desenvolvimento , Populus/metabolismo , Xilema/metabolismo , Xilema/genética , Xilema/crescimento & desenvolvimento , Câmbio/genética , Câmbio/crescimento & desenvolvimento , Câmbio/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Parede Celular/metabolismo , Proliferação de Células , Madeira/crescimento & desenvolvimento , Madeira/metabolismo , Madeira/genéticaRESUMO
Introduction: Until extended half-life (EHL) factor IX (FIX) concentrates became available in Japan in 2010, patients with hemophilia B received intravenous FIX replacement therapy with standard half-life (SHL) FIX concentrates. Purpose: To investigate the amount of factor dispensed and the associated medical expenditures for the treatment of hemophilia B in the real-world clinical setting in Japan. Methods: This retrospective study comprised patients with hemophilia B (N=197) who had filled prescriptions for FIX concentrates reported in Japan's Medical Data Vision database from 2015 to 2019. Patients were included if they had 2 or more prescriptions for the same FIX concentrates within the first 6 months of the study period and the interval between prescriptions was at least 2 weeks. Results: Since 2015, there was a decrease in the proportion of patients using SHL FIX concentrates and a corresponding increase in international units of dispensed EHL FIX concentrates. Median annualized dispensed dosages (IU/kg body weight) of EHL FIX concentrates were lower than for SHL concentrates for outpatient use only. Annual total health care expenditures per patient and annual expenditures for prescribed FIX concentrates increased each year during the study period. Following a switch from an SHL to an EHL concentrate, the median amount of prescribed FIX concentrate decreased slightly, although median total health care expenditures and FIX concentrate expenditures increased. Conclusion: In the real-world setting in Japan, medical expenditures and the proportion of patients prescribed EHL FIX concentrates for the treatment of hemophilia B have increased.
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Effective separation of lignin macromolecules from the xylose pre-hydrolysates (XPH) during the xylose production, thus optimizing the separation and purification process of xylose, is of great significance for reducing the production costs, achieving the high value-added utilization of lignin and increasing the industrial revenue. In this study, a simple and robust method (pH adjustment) for the separation of lignin from XPH was proposed and systematically compared with the conventional acid-promoted lignin precipitation method. The results showed that the lignin removal ratio (up to 60.34 %) of this simple method was higher than that of the conventional method, and the proposed method eliminated the necessity of heating and specialized equipment, which greatly reduced the separation cost. Meanwhile, this simple method does not destroy the components in XPH (especially xylose), ensuring the yield of the target product. On the other hand, the obtained lignin was nano-scale with less condensed structures, which also possessed small molecular weights with narrow distribution, excellent antioxidant activity (8-14 times higher than commercial antioxidants) and UV protection properties. In conclusion, the proposed simple separation method could effectively separate lignin from XPH at low cost, and the obtained lignin had potential commercial applications, which would further enhance the overall profitability of industrial production.
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Lignina , Xilose , Lignina/química , Xilose/química , Hidrólise , Bebidas AlcoólicasRESUMO
In this work, a green and robust pretreatment which integrated acetic acid-catalyzed hydrothermal and wet mechanical pretreatment, was developed to efficiently produce high yield (up to 40.12%) of xylooligosaccharides and digestible substrates from Caffeoyl Shikimate Esterase down-regulated and control poplar wood. Subsequently, superhigh yield (more than 95%) of glucose and residual lignin were obtained after a moderate enzymatic hydrolysis. The residual lignin fraction exhibited a well-preserved ß-O-4 linkages (42.06/100Ar) and high S/G ratio (6.42). Subsequently, lignin-derived porous carbon was successfully synthesized, and it exhibited a high specific capacitance of 273.8 F g-1 at 1.0 A g-1 and long cycling stability (remained 98.5% after 10,000 cycles at 5.0 A g-1) compared to control poplar wood, demonstrating that special advantage of this genetically-modified poplar in this integrated process. This work developed an energy-saving and eco-friendly pretreatment technology as a waste-free route for converting different lignocellulosic biomass to multiple products.
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Esterases , Lignina , Hidrólise , MadeiraRESUMO
Aim: To evaluate treatment patterns of novel therapies (inotuzumab ozogamicin (inotuzumab), blinatumomab, and tisagenlecleucel) in patients with acute lymphoblastic leukemia (ALL) in a Japanese real-world setting. Patients & Methods: Patients with ALL diagnoses from a Japanese claims database were examined. Results: We included 194 patients (97 patients were prescribed inotuzumab; 97 patients were prescribed blinatumomab; and no patient was prescribed tisagenlecleucel); 81.4% in the inotuzumab group and 78.4% in the blinatumomab group were prescribed chemotherapy prior to the initiation of those drugs. Most patients were prescribed subsequent treatment (60.8 and 58.8%, respectively). A small number of patients were prescribed sequential treatment of inotuzumab-to-blinatumomab or blinatumomab-to-inotuzumab (20.3 and 10.5%, respectively). Conclusion: This study revealed inotuzumab and blinatumomab treatment features in Japan.
In acute lymphoblastic leukemia (ALL), the increase in leukemic cells prevents the production of normal blood cells. As a result, people with ALL become more susceptible to anemia, fatigue, infections, fever, bruising and bleeding easily. ALL progresses rapidly without treatment. In recent years, new therapeutic drugs, including inotuzumab and blinatumomab, have become available; however, it remains unclear how they have been used in clinical practice. In this report, we assess how they are used in clinical practice using a large database to collect the clinical data of ALL patients. To see the treatment pattern, we found that most of the patients (81.4% of patients who received inotuzumab and 78.4% of those who received blinatumomab) had received chemotherapy before starting treatment with inotuzumab or blinatumomab. After patients ended treatment with inotuzumab or blinatumomab, 60.8% of patients who received inotuzumab and 58.8% of those who received blinatumomab received the next therapies, including chemotherapy. However, a small number of patients had received inotuzumab-to-blinatumomab or blinatumomab-to-inotuzumab (20.3 and 10.5%, respectively). These findings show the real-world treatment patterns of inotuzumab and blinatumomab; that is, both inotuzumab and blinatumomab are more likely to be prescribed to patients who might not have enough efficacy from prior chemotherapy or might have had to stop chemotherapy early due to side effects. Overall, there is clinical meaningful information from our findings of how inotuzumab and blinatumomab have been used for treatment of ALL and this information could improve the clinical practice of ALL in Japan.
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Anticorpos Biespecíficos , Leucemia-Linfoma Linfoblástico de Células Precursoras , Humanos , Japão/epidemiologia , Inotuzumab Ozogamicina/uso terapêutico , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Anticorpos Biespecíficos/efeitos adversos , Indução de RemissãoRESUMO
In this work, a rapid one-pot hydrated deep eutectic solvent (DES) pretreatment was proposed to facilitate the conversion of carbohydrates from lignocellulosic biomass to monosaccharides. Specifically, the pure and hydrated DES based on benzyl triethylammonium chloride (BTEAC), formic acid (FA) and water was used to pretreat bamboo shoot shells (BSS) by microwave heating. The pretreated solid residues were enzymatically saccharified to produce fermentable sugars, and the hydrolyzed carbohydrates and lignin remained in the hydrolyzate. The results showed that the yield of monosaccharides from the hydrated DES hydrolyzate (193.7-228.4 g/kg) was significantly higher than that (45.9-66.1 g/kg) of pure DES. The 30% hydrated DES pretreatment achieved the best glucose yield (89.03%) and a total monosaccharides yield of 555.4 g/kg, which corresponded to a conversion ratio of carbohydrates to monosaccharides of 87.0%. The proposed process is a robust method for the efficiently convert carbohydrates from BSS into monosaccharides.
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Carboidratos , Solventes Eutéticos Profundos , Lignina/química , Glucose/química , Monossacarídeos , Hidrólise , Biomassa , Solventes/químicaRESUMO
Cattle and buffalo served as the first and second largest dairy animals, respectively, providing 96% milk products worldwide. Understanding the mechanisms underlying milk synthesis is critical to develop the technique to improve milk production. Thiolases, also known as acetyl-coenzyme A acetyltransferases (ACAT), are an enzyme family that plays vital roles in lipid metabolism, including ACAT1, ACAT2, ACAA1, ACAA2, and HADHB. Our present study showed that these five members were orthologous in six livestock species including buffalo and cattle. Transcriptomic data analyses derived from different lactations stages showed that ACAA1 displayed different expression patterns between buffalo and cattle. Immunohistochemistry staining revealed that ACAA1 were dominantly located in the mammary epithelial cells of these two dairy animals. Knockdown of ACAA1 inhibited mammary epithelial cell proliferation and triglyceride and ß-casein secretion by regulating related gene expressions in cattle and buffalo. In contrast, ACAA1 overexpression promoted cell proliferation and triglyceride secretion. Finally, three novel SNPs (g.-681A>T, g.-23117C>T, and g.-24348G>T) were detected and showed significant association with milk production traits of Mediterranean buffaloes. In addition, g.-681A>T mutation located in the promoter region changed transcriptional activity significantly. Our findings suggested that ACAA1 play a key role in regulating buffalo and cattle milk synthesis and provided basic information to further understand the dairy animal lactation physiology.
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INTRODUCTION: A framework that extracts oncological outcomes from large-scale databases using artificial intelligence (AI) is not well established. Thus, we aimed to develop AI models to extract outcomes in patients with lung cancer using unstructured text data from electronic health records of multiple hospitals. METHODS: We constructed AI models (Bidirectional Encoder Representations from Transformers [BERT], Naïve Bayes, and Longformer) for tumor evaluation using the University of Miyazaki Hospital (UMH) database. This data included both structured and unstructured data from progress notes, radiology reports, and discharge summaries. The BERT model was applied to the Life Data Initiative (LDI) data set of six hospitals. Study outcomes included the performance of AI models and time to progression of disease (TTP) for each line of treatment based on the treatment response extracted by AI models. RESULTS: For the UMH data set, the BERT model exhibited higher precision accuracy compared to the Naïve Bayes or the Longformer models, respectively (precision [0.42 vs. 0.47 or 0.22], recall [0.63 vs. 0.46 or 0.33] and F1 scores [0.50 vs. 0.46 or 0.27]). When this BERT model was applied to LDI data, prediction accuracy remained quite similar. The Kaplan-Meier plots of TTP (months) showed similar trends for the first (median 14.9 [95% confidence interval 11.5, 21.1] and 16.8 [12.6, 21.8]), the second (7.8 [6.7, 10.7] and 7.8 [6.7, 10.7]), and the later lines of treatment for the predicted data by the BERT model and the manually curated data. CONCLUSION: We developed AI models to extract treatment responses in patients with lung cancer using a large EHR database; however, the model requires further improvement.
The use of artificial intelligence (AI) to derive health outcomes from large electronic health records is not well established. Thus, we built three different AI models: Bidirectional Encoder Representations from Transformers (BERT), Naïve Bayes, and Longformer to serve this purpose. Initially, we developed these models based on data from the University of Miyazaki Hospital (UMH) and later improved them using the Life Data Initiative (LDI) data set of six hospitals. The performance of the BERT model was better than the other two, and it showed similar results when it was applied to the LDI data set. The KaplanMeier plots of time to progression of disease for the predicted data by the BERT model showed similar trends to those for the manually curated data. In summary, we developed an AI model to extract health outcomes using a large electronic health database in this study; however, the performance of the AI model could be improved using more training data.
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Inteligência Artificial , Neoplasias Pulmonares , Humanos , Teorema de Bayes , População do Leste Asiático , Registros Eletrônicos de SaúdeRESUMO
Bamboo processing residue, which is rich in parenchyma cells, was treated as huge waste in bamboo processing industry, such as reassemble bamboo and bamboo flooring. Herein, autohydrolysis and rapid different deep eutectic solvents (DES) delignification strategy were consecutively performed to remove hemicelluloses and lignin from bamboo processing residue. The xylooligosaccharides (XOS) with high yield (34.35%) was achieved in the autohydrolysis process. Results showed that alkaline DES pretreatment resulted in the highest glucose yield (88.22%) and relatively high delignification rate (83.75%) as well as well-preserved lignin structures. However, the lignin fractions obtained under acidic DES conditions were tending to assemble into lignin nanoparticles (LNPs) and having excellent antioxidant activity as compared to those obtained from alkaline DES system. In brief, the combination of autohydrolysis and rapid DES delignification can achieve orientated fractionation of the components from the industrialized bamboo.
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Solventes Eutéticos Profundos , Lignina , Biomassa , Fracionamento Químico , Hidrólise , Lignina/química , Solventes/químicaRESUMO
An aspirational pretreatment method for efficient fractionation and tailored valorization of large industrial biomass can ensure the realizability of sustainable biorefinery strategies. In this study, an ultrafast alkaline deep eutectic solvents (DES) pretreatment strategy was developed to efficiently extract the lignin nanoparticles and retain cellulose residues that could be readily enzymatic saccharified to obtain fermentative glucose for the bioenergy production from industrial xylose residue. Results showed that the DES pretreatment had excellent delignification performance and the regenerated DES lignin nanoparticles exhibited well-preserved structures and excellent antioxidant activity, as well as low molecular weights and relatively uniform size distribution, which could facilitate downstream catalytic degradation for production of chemicals and preparation of lignin-based materials. Under the optimal condition (DES pretreatment: 80 °C, 10 min; saccharification: 10 FPU/g, 5 wt%, 100 mg/g Tween 80), the glucose yield of 90.12% could be achieved, which was dramatically increased compared to raw materials.
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Lignina , Xilose , Biomassa , Solventes Eutéticos Profundos , Glucose , Hidrólise , Lignina/química , Solventes/químicaRESUMO
A synergistic pretreatment that realizing effective fractionation and targeted valorization can guarantee the implementability to future biorefinery scenario. In the present study, a stepwise approach using hydrothermal and deep eutectic solvents (DES) pretreatment was developed to preferentially dissociate hemicelluloses and further remove lignin from poplar, while retaining a cellulose-rich substrate that can be easily digested via enzymatic saccharification to obtain glucose. Results showed that the hydrothermal filtrate is mainly composed of xylooligosaccharide (XOS), monosaccharides, byproducts, and xylan-type hemicelluloses, which have homogenous structures and uniform molecular weights distribution as well as excellent antioxidant activity. Subsequent DES pretreatment further removed the lignin barriers, leading to a remarkable increase in the saccharification efficiency from 15.72% to 96.33% under optimum conditions for enzymatic hydrolysis. In short, the integrated pretreatment is effective for dissociating and chemical conversion of poplar wood, which was reasonable to promote the frontier of highly available biorefinery.
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Celulose , Madeira , Biomassa , Hidrólise , Lignina , Polissacarídeos , SolventesRESUMO
Ethanol organosolv pretreatment is a green and effective deconstruction process for main components in lignocellulose biomass. Herein, balsa wood was firstly subjected to a modified ethanol/water solution (EWS) pretreatment with different Lewis acids catalysts (AlCl3, CuCl2, FeCl3) at 140-180 °C. The delignification ratios and structural characteristics of the dissociated lignin, enzymatic hydrolysis of cellulose in the pretreated substrates as well as the degradation products from hemicellulose during the pretreatment process were comprehensively investigated. Results showed that dissociation and depolymerization of lignin fragments was robust in AlCl3-catalyzed pretreatment than those by CuCl2 and FeCl3-catalyzed pretreatment. In detail, the results showed that the optimal delignification ratio and removal of the hemicelluloses occurred in AlCl3-catalyzed pretreatment. Moreover, the structural characterizations of lignin fractions by 2D-HSQC, 31P NMR and GPC also revealed that the obtained lignin has the advantages of small and homogeneous molecules as well as abundant functional groups. As a result of adequate removal of hemicellulose and lignin, the enzymatic digestibility of cellulose in the pretreated residue was significantly elevated. In short, the above findings are also in line with the concept of maximizing the utilization of bioresources, which will be beneficial for value-added applications of balsa wood in the biorefinery.
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Celulose/química , Etanol/química , Ácidos de Lewis/química , Lignina/química , Biomassa , Catálise , Espectroscopia de Ressonância MagnéticaRESUMO
Arthrobacter sp. B6 is a Gram-positive, non-motile, facultative aerobic bacterium, isolated from the arsenic-contaminated aquifer sediment in the Datong basin, China. This strain displays high resistance to arsenic, and can dynamically transform arsenic under aerobic condition. Here, we described the high quality draft genome sequence, annotations and the features of Arthrobacter sp. B6. The G + C content of the genome is 64.67%. This strain has a genome size of 4,663,437 bp; the genome is arranged in 8 scaffolds that contain 25 contigs. From the sequences, 3956 protein-coding genes, 264 pseudo genes and 89 tRNA/rRNA-encoding genes were identified. The genome analysis of this strain helps to better understand the mechanism by which the microbe efficiently tolerates arsenic in the arsenic-contaminated environment.
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Chondroitin sulfate (CS) plays important roles in the complement system. However, the CS structure is complicated due to different sources and the number and positions of sulfate groups. The objective of this study was to prepare different low molecular weight chondroitin sulfates (LMWCSs) and to investigate the biological activity in anti-complement capacity. A series of LMWCSs was prepared from different sources and characterized by ultraviolet-visible (UV) spectroscopy, high-performance liquid chromatography (HPLC), size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) and nuclear magnetic resonance (NMR) spectroscopy. Hemolytic, anti-complement 3 deposition capacity and cell viability assays were carried out to investigate the biological activities in vitro. The results showed that LMWCS prepared from shark cartilage with the oxidative degradation method (LMWCS-S-O) had the best anti-complement capacity. LMWCS-S-O could inhibit the alternative pathway of the complement system and protect chondrocytes from cell death. The attenuating effect of LMWCS-S-O on Osteoarthritis (OA) was investigated by destabilization of the medial meniscus (DMM) model in vivo. Functional wind-up, histological and C5b-9 analyses were used to evaluate the treatment effect on the OA model. In vivo results showed that LMWCS-S-O could attenuate OA. LMWCS-S-O with a high content of ΔDi-2,6diS and ΔDi-6S could be used for attenuating OA through regulating the complement system.
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Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacologia , Osteoartrite , Animais , Sobrevivência Celular/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Sulfatos de Condroitina/uso terapêutico , Modelos Animais de Doenças , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peso Molecular , Osteoartrite/tratamento farmacológico , Distribuição Aleatória , Resultado do TratamentoRESUMO
RNAi-based genetically engineered (GE) crops for the management of insect pests are likely to be commercialized by the end of this decade. Without a workable framework for conducting the ecological risk assessment (ERA) and a standardized ERA protocol, however, the utility of RNAi transgenic crops in pest management remains uncertain. The overall goal of this study is to assess the risks of RNAi-based GE crops on a non-target soil micro-arthropod, Sinella curviseta, which could be exposed to plant-protected dsRNAs deposited in crop residues. Based on the preliminary research, we hypothesized that insecticidal dsRNAs targeting at the western corn rootworm, Diabrotica virgifera virgifera, a billion-dollar insect pest, has no adverse impacts on S. curviseta, a soil decomposer. Following a tiered approach, we tested this risk hypothesis using a well-designed dietary RNAi toxicity assay. To create the worst-case scenario, the full-length cDNA of v-ATPase subunit A from S. curviseta were cloned and a 400 bp fragment representing the highest sequence similarity between target pest and non-target arthropods was selected as the template to synthesize insecticidal dsRNAs. Specifically, 10-days-old S. curviseta larvae were subjected to artificial diets containing v-ATPase A dsRNAs from both D. v. virgifera (dsDVV) and S. curviseta (dsSC), respectively, a dsRNA control, ß-glucuronidase, from plant (dsGUS), and a vehicle control, H2O. The endpoint measurements included gene expression profiles, survival, and life history traits, such as developmental time, fecundity, hatching rate, and body length. Although, S. curviseta larvae developed significantly faster under the treatments of dsDVV and dsSC than the vehicle control, the combined results from both temporal RNAi effect study and dietary RNAi toxicity assay support the risk hypothesis, suggesting that the impacts of ingested arthropod-active dsRNAs on this representative soil decomposer are negligible.
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This study reports the use of diethylenetriamine as background electrolyte for the simultaneous separation of hyaluronan acid, chondroitin sulfate, dermatan sulfate and heparin. The analytes were baseline separated by using an uncoated fused silica capillary at 37°C with a run time of 23min. The migration order, with hyaluronan acid at first and heparin at last, was related to the sulfation degree. The effect of salt concentration on resolution and migration order was also investigated. The developed method was applied to the simultaneous determination of hyaluronan acid and chondroitin sulfate in mouse plasma. Interferences in plasma were removed by protein precipitation and glycosaminoglycans were further purified by ethanol precipitation. The method was validated over the concentration range from 50 to 600µg/mL for hyaluronan acid and 500 to 6000µg/mL for chondroitin sulfate in mouse plasma. Results from assay validations showed that the method was selective and robust.
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Sulfatos de Condroitina/análise , Dermatan Sulfato/análise , Eletroforese Capilar/métodos , Heparina/análise , Ácido Hialurônico/análise , Plasma/química , Animais , Análise Química do Sangue/métodos , Camundongos , Poliaminas/químicaRESUMO
Few diazotrophs have been found to belong to the family Cytophagaceae so far. In the present study, a Gram-negative, rod-shaped bacterium that forms red colonies, was isolated from sands of the Takalamakan desert. It was designated H4XT. Phylogenetic and biochemical analysis indicated that the isolate is a new species of the genus Pontibacter. The 16S rRNA gene of H4XT displays 94.2-96.8% sequence similarities to those of other strains in Pontibacter. The major respiratory quinone is menaquinone-7 (MK-7). The DNA G+C content is 46.6 mol%. The major cellular fatty acids are iso-C15â¶0, C16â¶1ω5c, summed feature 3 (containing C16â¶1ω6c and/or C16â¶1ω7c) and summed feature 4 (comprising anteiso-C17â¶1B and/or iso-C17â¶1I). The major polar lipids are phosphatidylethanolamine (PE), one aminophospholipid (APL) and some unknown phospholipids (PLs). It is interesting to see that this bacterium can grow very well in a nitrogen-free medium. PCR amplification suggested that the bacterium possesses at least one type of nitrogenase gene. Acetylene reduction assay showed that H4XT actually possesses nitrogen-fixing activity. Therefore, it can be concluded that H4XT is a new diazotroph. We thus referred it to as Pontibacter diazotrophicus sp. nov. The type strain is H4XT (â=âCCTCC AB 2013049Tâ=âNRRL B-59974T).
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Cytophagaceae/classificação , Cytophagaceae/metabolismo , Ácidos Graxos/metabolismo , Técnicas de Tipagem Bacteriana , Cytophagaceae/genética , DNA Bacteriano/genética , Fixação de Nitrogênio , Filogenia , RNA Ribossômico 16S/genéticaRESUMO
A Gram-staining-negative, rod-shaped and pink bacterium was isolated from the soil of a Populus euphratica forest in the Taklamakan desert, Xinjiang, China. It was designated strain H9X(T). A 16S rRNA gene sequence homology search indicated that the isolate was most closely related to the family Cytophagaceae. The 16S rRNA gene of strain H9X(T) displayed 94.2-96.3â% sequence identities to those of type strains of other species of the genus Pontibacter. It only possessed menaquinone-7. The major cellular fatty acids of the novel isolate were iso-C15â:â0, C16â:â1ω5c summed feature 3 (containing C16â:â1ω6c and/or C16â:â1ω7c) and summed feature 4 (comprising anteiso-C17â:â1 B and/or iso-C17â:â1 I). The major polar lipids were phosphatidylethanolamine, one unknown aminophospholipid, one unknown glycophospholipid and several unknown phospholipids. The DNA G+C content of this bacterium was 55.2 mol%. Based on the phenotypic and genotypic data presented, it can be concluded that this isolate represents a novel species of the genus Pontibacter, for which the name Pontibacter yuliensis sp. nov. is proposed. The type strain is H9X(T) (â=âCCTCC AB 2013047(T)â=âKCTC 32396(T)).
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Cytophagaceae/classificação , Clima Desértico , Filogenia , Populus/microbiologia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Cytophagaceae/genética , Cytophagaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Dados de Sequência Molecular , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Árvores/microbiologia , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
The establishment of non-diazotrophic cultures of the filamentous marine cyanobacterium Trichodesmium erythraeum IMS101 enabled the first detailed investigation of the process leading to the development of its unique nitrogen-fixing cell type, the diazocyte. Trichome heterogeneity was apparent already within 3-8 h, while the differentiation of mature diazocytes, containing the nitrogenase enzyme, required 27 h after the removal of combined nitrogen. The distribution of 'pro-diazocytes' within the trichomes correlates with the localization of mature diazocytes, which suggests that pattern regulation is an early event during diazocyte development. The development was initially identified as changes in the subcellular ultrastructure, most notably the degradation of glycogen granules and gas vacuoles. These changes were preceded by the induced expression of the global nitrogen regulator ntcA at an early stage of combined nitrogen deprivation, followed by elevated expression of genes related to nitrogen metabolism and their corresponding proteins. The strongest induction (10-fold) was related to the transcription of the respiratory gene coxB2, apparent already at an early stage, which suggests an important role for respiration and the subsequent energy generation in the subcellular changes found, and in the creation of the reducing environment required for nitrogen fixation in diazocytes.