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Understanding the response of Chlorophyll a concentration (Chl-a, an indicator of phytoplankton biomass) to environmental factors is critical for eutrophication management. Light and nutrients often act as two main limiting environmental factors in large and shallow lakes. However, the limitation status is usually not considered explicitly when building empirical relationships even though the growth limitation is the possible mechanism controlling the behaviors of these relationships. Here we chose a typical large and shallow eutrophic lake (Lake Okeechobee) to study the response of Chl-a concentration under different growth limitation conditions. Using an existed decision tree model followed by Carlson's trophic state index, monitoring data from 1994 to 2020 were classified into light-limitation, nitrogen-limitation, or phosphorus-limitation. The spatio-temporal patterns of limitation status were revealed. By subdivision of observations according to these growth limitation classes, our results demonstrated three main findings. First, algae responded differently between light limitation and nutrient limitation. Chl-a concentrations were lower with smaller variability when light was limiting than those when nutrient was limiting. In addition, the evolution of Chl-a in enduring nutrient limitation events were more dynamic. Second, limitation-specific regressions provided a more straightforward interpretation compared with those without consideration of limitation status. Chl-a â¼ nutrient relationship based on limitation classification displayed a higher R2 with a positive slope. This positive slope indicates the sensitivity of Chl-a to that specific nutrient. Moreover, response of Chl-a to phosphorus was successfully detected by identifying P-limited samples. Otherwise, the Chl-a â¼ TP response would be muted since nitrogen is the main limiting nutrient in Lake Okeechobee. Third, a spatial heterogeneity of Chl-a â¼ TN relationship was revealed by Bayesian hierarchical modelling. This indicates the necessity of focusing more on hot spots where Chl-a displays a higher sensitivity to increase of nutrient. Our findings demonstrate the advantage of developing the limitation-specific and zone-specific relationships between algal biomass and environmental factors.
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Clorofila , Lagos , Teorema de Bayes , Clorofila A , EutrofizaçãoRESUMO
BACKGROUND: Nanomaterials in plant protection promise many benefits over conventional pesticide products. Nano-enabled pesticides may alter the functionality or risk profile of active ingredients. Cationic nanochitin whiskers (NC) possess strong biological activity against wheat aphids. However, toxicity and synergistic effects of NC with chemical pesticides against pest insects has not been systemically reported. This study investigated the insecticidal enhancement by NC with Omethoate (40% EC), Imidacloprid (10% WP), and Acetamiprid (40% WG) for pest control using wheat aphid as piercing-sucking mouthparts insect. Fluorescein isothiocyanate labelled NC was used to monitor the uptake and transportation pathway of NC inside the target insects. Toxicity of NC was tested with Sprague-Dawley (SD) rat. Our findings provide a theoretical basis for future application of NC in plant protection against pest insects. RESULTS: NCs synthesized by acidic hydrolysis were rod-like nanoparticles in a range of 50-150 nm in length and 30-50 nm in width, which examined by electron microscopy and dynamic light scattering methods. The charge density and zeta potential were about 63 mmol/kg and + 36.4 mV, respectively. By absorption and/or contact action of 30-50 mg/L of NC suspension, the corrected mortality of wheat aphids reached up to 80% or above after 12 h treatment, NC could be distributed through digestive system and relocated from mouth to other tissues inside the insect body. When associated with dilutions of conventional pesticides, the corrected mortality were significantly increased up to 95% or above. The dosage of the chemical pesticide and nanochitin in the mixtures (1:1 by volume) were all reduced to half. The acute oral toxicity Lethal Dose 50% (LD50) to SD rat is greater than 5000 mg/kg BW (body weight) in male and female, acute dermal toxicity LD50 is greater than 2000 mg/kg BW of NC. CONCLUSIONS: NC has a strong promotive effect on insecticidal effectiveness of chemical insecticides. It was easily absorbed by plant, transported and distributed from mouth to other tissues of the insects while sucking plant fluid. Low acute oral and dermal toxicity to SD rat indicated that it is safe to apply in agriculture and food industry. NCs has a great potential for water-based nanopesticide formulation to reduce chemical pesticide use for future agro-environmental sustainability.
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Quitina/química , Inseticidas/farmacologia , Vibrissas/química , Animais , Afídeos , Feminino , Controle de Insetos , Masculino , Camundongos , Mortalidade , Neonicotinoides , Nitrocompostos , Praguicidas , Ratos , Ratos Sprague-Dawley , TriticumRESUMO
BACKGROUND: Myocardial ischemia-reperfusion (MI/R) injury is a significant clinical problem without effective therapy. Unbiased omics approaches may reveal key MI/R mediators to initiate MI/R injury. METHODS: We used a dynamic transcriptome analysis of mouse heart exposed to various MI/R periods to identify S100a8/a9 as an early mediator. Using loss/gain-of-function approaches to understand the role of S100a8/a9 in MI/R injury, we explored the mechanisms through transcriptome and functional experiment. Dynamic serum S100a8/a9 levels were measured in patients with acute myocardial infarction before and after percutaneous coronary intervention. Patients were prospectively followed for the occurrence of major adverse cardiovascular events. RESULTS: S100a8/a9 was identified as the most significantly upregulated gene during the early reperfusion stage. Knockout of S100a9 markedly decreased cardiomyocyte death and improved heart function, whereas hematopoietic overexpression of S100a9 exacerbated MI/R injury. Transcriptome/functional studies revealed that S100a8/a9 caused mitochondrial respiratory dysfunction in cardiomyocytes. Mechanistically, S100a8/a9 downregulated NDUF gene expression with subsequent mitochondrial complex I inhibition via Toll-like receptor 4/Erk-mediated Pparg coactivator 1 alpha/nuclear respiratory factor 1 signaling suppression. Administration of S100a9 neutralizing antibody significantly reduced MI/R injury and improved cardiac function. Finally, we demonstrated that serum S100a8/a9 levels were significantly increased 1 day after percutaneous coronary intervention in patients with acute myocardial infarction, and elevated S100a8/a9 levels were associated with the incidence of major adverse cardiovascular events. CONCLUSIONS: Our study identified S100a8/a9 as a master regulator causing cardiomyocyte death in the early stage of MI/R injury via the suppression of mitochondrial function. Targeting S100a8/a9-intiated signaling may represent a novel therapeutic intervention against MI/R injury. CLINICAL TRIAL REGISTRATION: URL: https://www.clinicaltrials.gov. Unique identifier: NCT03752515.
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Apoptose , Calgranulina B/metabolismo , Mitocôndrias/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Animais , Anticorpos Neutralizantes/administração & dosagem , Calgranulina A/sangue , Calgranulina B/genética , Calgranulina B/imunologia , Modelos Animais de Doenças , Complexo I de Transporte de Elétrons/antagonistas & inibidores , Complexo I de Transporte de Elétrons/metabolismo , Insuficiência Cardíaca/etiologia , Humanos , Potencial da Membrana Mitocondrial , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Infarto do Miocárdio/diagnóstico , Infarto do Miocárdio/cirurgia , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , Intervenção Coronária Percutânea , Transdução de SinaisRESUMO
BACKGROUND: Hypertensive ventricular remodeling is a common cause of heart failure. However, the molecular mechanisms regulating ventricular remodeling remain poorly understood. METHODS: We used a discovery-driven/nonbiased approach to identify increased activating transcription factor 3 (ATF3) expression in hypertensive heart. We used loss/gain of function approaches to understand the role of ATF3 in heart failure. We also examined the mechanisms through transcriptome, chromatin immunoprecipitation sequencing analysis, and in vivo and in vitro experiments. RESULTS: ATF3 expression increased in murine hypertensive heart and human hypertrophic heart. Cardiac fibroblast cells are the primary cell type expressing high ATF3 levels in response to hypertensive stimuli. ATF3 knockout (ATF3KO) markedly exaggerated hypertensive ventricular remodeling, a state rescued by lentivirus-mediated/miRNA-aided cardiac fibroblast-selective ATF3 overexpression. Conversely, conditional cardiac fibroblast cell-specific ATF3 transgenic overexpression significantly ameliorated ventricular remodeling and heart failure. We identified Map2K3 as a novel ATF3 target. ATF3 binds with the Map2K3 promoter, recruiting HDAC1, resulting in Map2K3 gene-associated histone deacetylation, thereby inhibiting Map2K3 expression. Genetic Map2K3 knockdown rescued the profibrotic/hypertrophic phenotype in ATF3KO cells. Last, we demonstrated that p38 is the downstream molecule of Map2K3 mediating the profibrotic/hypertrophic effects in ATF3KO animals. Inhibition of p38 signaling reduced transforming growth factor-ß signaling-related profibrotic and hypertrophic gene expression, and blocked exaggerated cardiac remodeling in ATF3KO cells. CONCLUSIONS: Our study provides the first evidence that ATF3 upregulation in cardiac fibroblasts in response to hypertensive stimuli protects the heart by suppressing Map2K3 expression and subsequent p38-transforming growth factor-ß signaling. These results suggest that positive modulation of cardiac fibroblast ATF3 may represent a novel therapeutic approach against hypertensive cardiac remodeling.
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Fator 3 Ativador da Transcrição/metabolismo , Fibroblastos/enzimologia , Insuficiência Cardíaca/prevenção & controle , Hipertrofia Ventricular Esquerda/prevenção & controle , MAP Quinase Quinase 3/metabolismo , Miocárdio/enzimologia , Função Ventricular Esquerda , Remodelação Ventricular , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Acetilação , Fator 3 Ativador da Transcrição/deficiência , Fator 3 Ativador da Transcrição/genética , Angiotensina II , Animais , Sítios de Ligação , Células Cultivadas , Modelos Animais de Doenças , Fibroblastos/patologia , Fibrose , Predisposição Genética para Doença , Insuficiência Cardíaca/enzimologia , Insuficiência Cardíaca/etiologia , Insuficiência Cardíaca/fisiopatologia , Histona Desacetilase 1/metabolismo , Histonas/metabolismo , Humanos , Hipertensão/induzido quimicamente , Hipertrofia Ventricular Esquerda/enzimologia , Hipertrofia Ventricular Esquerda/etiologia , Hipertrofia Ventricular Esquerda/fisiopatologia , MAP Quinase Quinase 3/genética , Masculino , Camundongos Knockout , Miocárdio/patologia , Fenótipo , Regiões Promotoras Genéticas , Inibidores de Proteínas Quinases/farmacologia , Transdução de Sinais , Fatores de Tempo , Fator de Crescimento Transformador beta/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidoresRESUMO
PURPOSE: We sought to determine the impact of levosimendan on mortality following cardiac surgery based on large-scale randomized controlled trials (RCTs). METHODS: We searched PubMed, Web of Science, Cochrane databases, and ClinicalTrials.gov for RCTs published up to December 2017, on levosimendan for patients undergoing cardiac surgery. RESULTS: A total of 25 RCTs enrolling 2960 patients met the inclusion criteria; data from 15 placebo-controlled randomized trials were included for meta-analysis. Pooled analysis showed that the all-cause mortality rate was 6.4% (71 of 1106) in the levosimendan group and 8.4% (93 of 1108) in the placebo group (odds ratio [OR], 0.76; 95% confidence interval [CI], 0.55-1.04; P = 0.09). There were no significant differences between the two groups in the rates of myocardial infarction (OR: 0.91; 95% CI, 0.68-1.21; P = 0.52), serious adverse events (OR: 0.84; 95% CI, 0.66-1.07; P = 0.17), hypotension (OR: 1.69; 95% CI, 0.94-3.03; P = 0.08), and low cardiac output syndrome (OR: 0.47; 95% CI, 0.22-1.02; P = 0.05). CONCLUSION: Levosimendan did not result in a reduction in mortality in adult cardiac surgery patients. Well designed, adequately powered, multicenter trials are necessary to determine the role of levosimendan in adult cardiac surgery.
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Baixo Débito Cardíaco/mortalidade , Baixo Débito Cardíaco/prevenção & controle , Procedimentos Cirúrgicos Cardíacos , Bases de Dados Bibliográficas , Hidrazonas/administração & dosagem , Inibidores de Fosfodiesterase/administração & dosagem , Complicações Pós-Operatórias/mortalidade , Complicações Pós-Operatórias/prevenção & controle , Piridazinas/administração & dosagem , Ensaios Clínicos Controlados Aleatórios como Assunto , Humanos , SimendanaRESUMO
OBJECTIVE: Interleukin (IL)-1ß and IL-18 are key proinflammatory cytokines that play important roles in the pathophysiology of vein graft remodeling. However, the mechanism of IL-1ß/IL-18 production and its role in the development of graft remodeling remain unclear. APPROACH AND RESULTS: IL-1ß/IL-18 were rapidly expressed in venous interposition grafts. Vascular smooth muscle cell (VSMC) death and monocytic inflammasome activation occurred in grafted veins. Necrotic VSMCs induced the expression of IL-1ß, IL-18, and other inflammasome-associated proteins in monocytes, which was partially inhibited by their antagonist, recombinant IL-1ra-Fc-IL-18bp. Activated monocytes stimulated proliferation of VSMCs by activating cell growth-related signaling molecules (AKT, STAT3, ERK1/2, and mTOR [AKT/protein kinase B, signal transducer and activator of transcription 3, extracellular signal-regulated kinase 1/2, mammalian target of rapamycin]) and increasing production of platelet-derived growth factor-bb; these effects were suppressed by IL-1ra-Fc-IL-18bp. Activated monocytes also promoted migration of VSMCs, which was independent of IL-1ß/IL-18 signaling. Importantly, administration of IL-1ra-Fc-IL-18bp inhibited activation of cell growth-related signaling molecules, VSMC proliferation, and vein graft thickening in vivo. CONCLUSIONS: Our work identified an interaction among necrotic VSMCs, monocytes, and viable VSMCs through IL-1ß/IL-18 signaling, which might be exploited as a therapeutic target in vein graft remodeling.
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Implante de Prótese Vascular , Artérias Carótidas/cirurgia , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Proteína Antagonista do Receptor de Interleucina 1/farmacologia , Interleucina-18/fisiologia , Interleucina-1beta/fisiologia , Monócitos/citologia , Músculo Liso Vascular/citologia , Miócitos de Músculo Liso/citologia , Neointima , Proteínas Recombinantes de Fusão/farmacologia , Transdução de Sinais/fisiologia , Veia Cava Superior/transplante , Animais , Apoptose , Linhagem Celular Tumoral , Células Cultivadas , Meios de Cultivo Condicionados/farmacologia , Humanos , Inflamassomos/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/uso terapêutico , Proteína Antagonista do Receptor de Interleucina 1/uso terapêutico , Interleucina-18/biossíntese , Interleucina-18/genética , Interleucina-1beta/biossíntese , Interleucina-1beta/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Monócitos/metabolismo , Miócitos de Músculo Liso/metabolismo , Necrose , RNA Mensageiro/biossíntese , Proteínas Recombinantes de Fusão/uso terapêutico , Veia Safena/citologia , Organismos Livres de Patógenos Específicos , Veia Cava Superior/metabolismoRESUMO
Hydrophobic, monodisperse LaPO4: Ce³âº nanoparticles, LaPO4:Ce³âº/LaPO4 and LaPO4:Ce³âº/LaPO4: Ce³âº/ LaPO4 core/shell structure nanocrystals were synthesized via a high-temperature organic solution approach. The as-synthesized samples were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR) and photoluminescence spectroscopy (PL). The results show that: all the samples are a monoclinic phase, Because of the size small nanoparticles, the diffraction peaks of the sample occurs width phenomenon. The LaPO4:Ce³âº nano- crystals exhibits the characteristic emission of Ce³âº ions, the photoluminescence intensity increases first and then decreases with the increasing doping concentration of Ce³âº ions, and the best doping amount is 6 at %, with the increasing doping amount, the photoluminescence intensity decreases which may caused by the concentration quenching. Compared to LaPO4:Ce³âº nanoparti- cles, the photoluminescence intensity of LaPO4:Ce³âº/LaPO4 and LaPO4: Ce³âº/LaPO4: Ce³âº/LaPO4 core/shell structure nanocrystals improves about 41% and 95% respectively, this may be a synergy of larger particle size of nanocrystals and surface passivation effect. FTIR spectra data shows that the absorption peak at 1545 and 1461 cm⻹ corresponded to the asymmetric and symmetric stretching vibration of --COOâ», the separation, Δ, between the two peaks is 84 cm⻹, The mechanism of the sample surface modification by the organics might be that the oxygen atoms of the carboxyl are coordinated with the lanthunum ions by a bidentate mode.
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Eusocial insects have evolved specific defensive strategies to protect their colonies. In termite colonies, soldiers perform a colony-level defense by displaying mechanical biting, head-banging and mandible opening-closing behaviors. However, few studies have been reported on the factors modulating defensive behaviors in termites. Owing to JH (juvenile hormone) being involved in soldier differentiation, JH was speculated to affect defensive behaviors in termite soldiers. To determine the effect of JH on the defensive behaviors of termite soldiers, we performed a JHA-feeding and RaSsp1-silencing experiment and then tested the changes in defense-related behaviors, alarm pheromones and key JH signaling genes. The observed result was that after feeding workers with JHA, soldiers displayed the following: (1) decreased biting events and increased head-banging events; (2) a reduced expression of RaSsp1 and increased expression of Met (methoprene-tolerant, the nuclear receptor of JH) and Kr-h1 (the JH-inducible transcription factor Krüppel homolog 1); and (3) a decreased concentration of alarm pheromones, including α-pinene, ß-pinene and limonene (+, -). Further study showed that soldiers silenced for RaSsp1 also exhibited (1) decreased biting events and increased head-banging events and (2) increased expression of Met and Kr-h1. In addition, soldiers stimulated by the alarm pheromone limonene displayed an increase in the frequency of mandible opening-closing and biting behavior. All of these results show that JHA influenced the defensive behaviors of termite soldiers, possibly via downregulating RaSsp1 expression, up-regulating Met and Kr-h1 and stimulating the secretion of alarm pheromones, suggesting that the JH pathway plays important roles in modulating social behaviors in termite colonies.
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Acyl-coenzyme A:cholesterol acyltransferase 2 (ACAT2) is important for cholesterol ester synthesis and secretion. A previous study revealed that ACAT2 gene promoter activity was upregulated by hepatocyte nuclear factor 4α (HNF4α) through two sites around -247 and -311 of ACAT2 gene promoter. Here, we identified two novel cis-elements, site I (-1006 to -898) and site II (-38 to -29), which are important for HNF4α effect. In HepG2 cells, mutation of site I decreased ACAT2 gene promoter activity to one-fifth of that of the wild type, while mutation of site II reduced promoter activity to less than one-tenth of that of the wild type. In 293T cells, mutation of these two cis-elements profoundly impaired the HNF4α induction effect. When either of these two elements was inserted into pGL3-promoter, HNF4α induced promoter activity through the inserted element, while mutation of the element impaired HNF4α induction effect. In electrophoretic mobility shift assay and chromatin immunoprecipitation experiment, HNF4α bound to these two elements. Thus, the two cis-elements are important for HNF4α effect on ACAT2 gene transcription. We also showed that HNF4α positively regulates ACAT2 gene expression at mRNA level. Overexpression of HNF4α increased ACAT2 expression, whereas knockdown of HNF4α decreased ACAT2 expression. Peroxisome proliferator-activated receptor gamma coactivator 1α (PCG1α), a coactivator of HNF4α, increased ACAT2 expression, while small heterodimer partner (SHP), a corepressor of HNF4α, decreased ACAT2 expression. These results provide more insights into transcriptional regulation of ACAT2 expression.
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Fator 4 Nuclear de Hepatócito/metabolismo , Esterol O-Aciltransferase/genética , Sequência de Bases , Sítios de Ligação/genética , Linhagem Celular , Primers do DNA/genética , Regulação Enzimológica da Expressão Gênica , Técnicas de Silenciamento de Genes , Teste de Complementação Genética , Células HeLa , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Células Hep G2 , Fator 4 Nuclear de Hepatócito/antagonistas & inibidores , Fator 4 Nuclear de Hepatócito/genética , Humanos , Mutação , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Regiões Promotoras Genéticas , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Esterol O-Aciltransferase 2RESUMO
Highly enantioselective rhodium-catalyzed hydrogenation of (Z)-N-sulfonyl-α-dehydroamido boronic esters is realized for the first time using a JosiPhos-type ligand. This method has enabled convenient synthesis of a series of enantio-enriched N-sulfonyl-α-amido boronic esters in good yields and excellent enantioselectivities (up to 99% ee).
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Exploring an efficient and green pretreatment method is an important prerequisite for the development of biorefinery. It is well known that locusts can degrade gramineous lignocellulose efficiently. Locusts can be used as a potential resource for studying plant cell wall degradation, but there are few relative studies about locusts so far. Herein, some new discoveries were revealed about elucidating the process of biodegradation of gramineous lignocellulose in Locusta migratoria manilensis. The enzyme activity related to lignocellulose degradation and the content of cellulose, hemicellulose, and lignin in the different gut segments of locusts fed corn leaves were measured in this study. A series of characterization analyses were conducted on corn leaves and locust feces, which included field emission scanning electron microscopy (FE-SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction pattern (XRD), and thermogravimetric (TG) analysis. These results showed that the highest activities of carboxymethyl cellulase (CMCase), filter paper cellulase (FPA), and xylanase were obtained in the foregut of locusts, which strongly indicated that the foregut was the main lignocellulose degradation segment in locusts; furthermore, the majority of nutritional components were absorbed in the midgut of locusts. The activity of CMCase was significantly higher than that of xylanase, and manganese peroxidase (MnPase) activity was lowest, which might be due to the basic nutrition of locusts being cellulose and hemicellulose and not lignin based on the results of FE-SEM, FTIR, XRD, and TG analysis. Overall, these results provided a valuable insight into lignocellulosic degradation mechanisms for understanding gramineous plant cell wall deconstruction and recalcitrance in locusts, which could be useful in the development of new enzymatic pretreatment processes mimicking the locust digestive system for the biochemical conversion of lignocellulosic biomass to fuels and chemicals.
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Termite soldiers are a unique caste among social insects, and their differentiation can be induced by Juvenile hormone (JH) from workers through two molts (worker-presoldier-soldier). However, the molecular mechanism underlying the worker-to-soldier transformation in termites is poorly understood. To explore the mechanism of soldier differentiation induced by JH, the gene soldier caste-specific protein 1 (RaSsp1, NCBI accession no: MT861054.1) in R. aculabialis was cloned, and its function was studied. This gene was highly expressed in the soldier caste, and the protein RsSsp1 was similar to the JHBP (JH-binding protein) domain-containing protein by Predict Protein online. In addition, JHIII could be anchored in the hydrophobic cage of RaSsp1 as the epoxide of the JHBP-bound JH according to the protein ligand molecular docking online tool AutoDock. The functional studies indicated that knocking down of the RaSsp1 shorted the presoldier's head capsule, reduced mandible size, delayed molting time and decreased molting rate (from worker to presoldier) at the beginning of worker gut-purging. Furthermore, knocking down of the RaSsp1 had a more pronounced effect on soldier differentiation (from presoldier to soldier), and manifested in significantly shorter mandibles, rounder head capsules, and lower molting rate (from worker to presoldier) at the beginning of presoldier gut-purging. Correspondingly, the expressions of JH receptor Methoprene-tolerant (Met), the JH-inducible transcription factor Krüppel homolog1 (Kr-h1) and ecdysone signal genes Broad-complex (Br-C) were downregulated when knocking down the RaSsp1 at the above two stages. All these results that RaSsp1 may be involved in soldier differentiation from workers by binding and transporting JH.
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A modified Perkow reaction, named Perkow-Shi reaction, was developed based on the one-pot α-tosyloxylation of ketones following by addition of P(iii)-reagents and 4 Å molecular sieves. Diversity of enol phosphates, as well as enol phosphonates, enol phosphinates, and enol phosphoramidates were synthesized in high yields directly from the ubiquitously available ketones instead of the unfavourable α-chloroketones under a mild and environmental friendly condition.
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BACKGROUND & AIMS: miR-122 is the most abundant microRNA in the liver and regulates metabolic pathways including cholesterol biosynthesis, fatty acid synthesis, and oxidation. However, little is known about mechanisms that regulate the expression of miR-122 in the liver. The aim of this study was to identify key transcriptional regulators for miR-122 expression through intensively studying its primary transcript and promoter region. METHODS: Bioinformatics analysis, Northern blotting, RT-PCR, and 5'/3' RACE were performed to analyze miR-122 primary transcript structure, its promoter region, and potential transacting factor binding sites. Reporter gene assays integrated with truncation and site-mutation in miR-122 promoter were performed to determine the trans-activation effect of HNF4α to miR-122-promoter in vitro. ChIP and EMSA assays were performed to determine HNF4α binding to miR-122 promoter. Finally, forced expression and RNAi were performed to verify the regulatory roles of HNF4 to miR-122 expression in vitro and in vivo. RESULTS: Here, we show that miR-122 is processed from a long spliced primary transcript directed by a distal upstream promoter region conserved across species. We dissected this promoter region and identified putative binding sites for liver-enriched transcriptional factors that contribute to the regulation of miR-122 expression, including a putative binding site for hepatocyte nuclear factor 4α (HNF4α). We demonstrate that HNF4α binds to the miR-122 promoter region through the conserved DR-I element. We observed the DR-1-element-dependent activation effect of HNF4α on the conserved miR-122 promoter and the activation could be further enhanced by the addition of PGC1α. Using overexpression and knockdown strategies, we show that HNF4α positively regulates miR122 expression in both Huh7 cells and the mouse liver. CONCLUSIONS: Our results suggest that HNF4α is a key regulator of miR-122 expression in the liver.
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Regulação da Expressão Gênica , Fator 4 Nuclear de Hepatócito/metabolismo , MicroRNAs/genética , Fatores de Transcrição/metabolismo , Transcrição Gênica , Animais , Imunoprecipitação da Cromatina , Ensaio de Desvio de Mobilidade Eletroforética , Células HeLa , Células Hep G2 , Fator 4 Nuclear de Hepatócito/genética , Fator 4 Nuclear de Hepatócito/fisiologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , MicroRNAs/metabolismo , Regiões Promotoras Genéticas , Análise de Sequência de DNA , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologiaRESUMO
Substance P, an 11-residue neuropeptide, can be rapidly internalized through specific interaction with the neurokinin-1 receptor. Therefore, we designed and synthesized the substance P targeted camptothecin (CPT) conjugates via a releasable disulfide carbonate linker. All the conjugates exhibited comparable or stronger cytotoxicity to cancer cells that highly over-express neurokinin-1 receptor than free CPT. More importantly, the selectivity of conjugates was significantly improved compared with CPT. Our results indicated that these conjugates can be promising candidates for new chemotherapeutic drugs. In addition, increasing CPT loading or attachment of CPT to the C-terminal hexapeptide of substance P are useful strategies to enhance the therapeutic efficacy of substance P targeted conjugates.
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Antineoplásicos/síntese química , Camptotecina/síntese química , Sistemas de Liberação de Medicamentos , Substância P/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Camptotecina/química , Camptotecina/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Preparações de Ação Retardada , Humanos , Estrutura Molecular , Neoplasias , Neurotransmissores/químicaRESUMO
Mycoplasma hyopneumoniae causes swine respiratory disease worldwide. Due to the difficulty of isolating and cultivating M. hyopneumoniae, very few attenuated strains have been successfully isolated, which hampers the development of attenuated vaccines. In order to produce an attenuated M. hyopneumoniae strain, we used the highly virulent M. hyopneumoniae strain ES-2, which was serially passaged in vitro 200 times to produce the attenuated strain ES-2L, and its virulence was evidenced to be low in an animal experiment. In order to elucidate the mechanisms underlying virulence attenuation, we performed whole-genome sequencing of both strains and conducted comparative genomic analyses of strain ES-2 and its attenuated form ES-2L. Strain ES-2L showed three large fragment deletion regions including a total of 18 deleted genes, compared with strain ES-2. Analysis of single-nucleotide polymorphisms (SNPs) and indels indicated that 22 dels were located in 19 predicted coding sequences. In addition to these indels, 348 single-nucleotide variations (SNVs) were identified between strains ES-2L and ES-2. These SNVs mapped to 99 genes where they appeared to induce amino acid substitutions and translation stops. The deleted genes and SNVs may be associated with decreased virulence of strain ES-2L. Our work provides a foundation for further examining virulence factors of M. hyopneumoniae and for the development of attenuated vaccines.
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Carabidae are one of the most species-rich families of beetles, comprising more than 40,000 described species worldwide. Forty-three complete or partial mitochondrial genomes (mitogenomes) from this family have been published in GenBank to date. In this study, we sequenced a nearly complete mitogenome of Amara aulica (Carabidae), using a next-generation sequencing method. This mitogenome was 16,646 bp in length, which encoded the typical 13 mitochondrial protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a putative control region. Combining with the published mitogenomes of Carabidae and five outgroup species from Trachypachidae, Gyrinidae and Dytiscidae, we performed phylogenetic estimates under maximum likelihood and Bayesian inference criteria to investigate the phylogenetic relationships of carabid beetles. The results showed that the family Carabidae was a non-monophyletic assemblage. The subfamilies Cicindelinae, Elaphrinae, Carabinae, Trechinae and Harpalinae were recovered as monophyletic groups. Moreover, the clade (Trechinae + (Brachininae + Harpalinae)) was consistently recovered in all analyses.
Assuntos
Besouros/genética , Genoma Mitocondrial , Mitocôndrias/genética , Animais , Teorema de Bayes , Besouros/classificação , Genes de RNAr , Sequenciamento de Nucleotídeos em Larga Escala , Mitocôndrias/metabolismo , Proteínas Mitocondriais/genética , Filogenia , RNA de Transferência/genética , Alinhamento de SequênciaRESUMO
Aberrant expression of microRNAs (miRNAs) in various human cancers suggests a role for miRNAs in tumorigenesis. Dicer is an essential component of the miRNA machinery, which mediates the processing of miRNAs. However, little is known about the role of Dicer in tumor proliferation and drug resistance. In this study, we found that knockdown of Dicer by siRNA led to significant G1 arrest and increased sensitivity to the DNA damaging agent, cisplatin, in breast cancer cell line MCF-7. Moreover, we found down-regulation of miR-21, a well-recognized miRNA frequently involved in a wide variety of cancers and up-regulation of cell cycle-dependent kinase inhibitor (CKI) p21 and p27. These data demonstrate that knockdown of Dicer inhibits human breast carcinoma cell growth and suggests a promising combination of anti-Dicer strategy and traditional chemotherapy to improve cancer treatment efficiency.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/enzimologia , Cisplatino/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Ribonuclease III/metabolismo , Western Blotting , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Feminino , Fase G1/genética , Humanos , MicroRNAs/metabolismo , RNA Interferente Pequeno , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ribonuclease III/genética , TransfecçãoRESUMO
Polycrystalline SrRu(1-x)Mn(x)O(3) (0≤x≤1) perovskite oxides have been prepared by a conventional solid-state reaction technique. Magnetic and magnetotransport properties are measured using a superconducting quantum interference device (SQUID, Quantum Design MPMS) over a temperature range of 4-300 K. The substitution of Mn ions for Ru drives the system from a ferromagnetic state, SrRuO(3), to an antiferromagnetic state, SrMnO(3), which is basically similar to observations in single-crystal SrRu(1-x)Mn(x)O(3) (Cao et al 2005 Phys. Rev. B 71 035104). However, the measurement of dc magnetization and ac susceptibility indicates that magnetic phase transition with x is more complicated and pronounced than those in single crystals. The phase transition process as temperature is reduced covers paramagnetism-antiferromagnetism (PM-AFM), paramagnetism-ferromagnetism (PM-FM) and ferromagnetism-cluster glass-spin glass (FM/CG/SG) etc. In particular, we observe a large low-temperature magnetoresistance (MR) of -41% for the sample x = 0.55, which is the largest MR measured in Mn-doped SrRuO(3). The experiment has verified that the large MR stems predominantly from a unique spin glass state in the polycrystalline alloy. These results substantiate that Ru-based oxides doped with 3d/4d transition metals have the potential for use in spintronics devices due to their adjustable phase transition, depending upon the level and nature of 3d/4d ion doping.
RESUMO
Nonsense-mediated RNA decay (NMD), an mRNA quality control mechanism, triggers degradation of mRNAs that contain premature termination codon (PTC) within their coding regions. NMD is a relatively conservative process that involves many trans-acting factors. The key domains for their function in NMD are conserved in evolution. These trans-acting factors are classified as different groups by their function in NMD. In addition, the mRNP formation is dynamic in NMD process because of sequential recruitment and interaction of these factors. To gain an insight into the mechanism of NMD, we dissect the mechanism of NMD based on the information on the structure, the regulation and interaction of these factors.