RESUMEN
MAIN CONCLUSION: The Cas13a-based multiplex RNA targeting system can be engineered to confer resistance to RNA viruses, whereas the number and expression levels of gRNAs have no significant effect on viral interference. The CRISPR-Cas systems provide adaptive immunity to bacterial and archaeal species against invading phages and foreign plasmids. The class 2 type VI CRISPR/Cas effector Cas13a has been harnessed to confer the protection against RNA viruses in diverse eukaryotic species. However, whether the number and expression levels of guide RNAs (gRNAs) have effects on the efficiency of RNA virus inhibition is unknown. Here, we repurpose CRISPR/Cas13a in combination with an endogenous tRNA-processing system (polycistronic tRNA-gRNA) to target four genes of potato virus Y (PVY) with varying expression levels. We expressed Cas13a and four different gRNAs in potato lines, and the transgenic plants expressing multiple gRNAs displayed similar suppression of PVY accumulation and reduced disease symptoms as those expressing a single gRNA. Moreover, PTG/Cas13a-transformed plants with different expression levels of multiple gRNAs displayed similar resistance to PVY strains. Collectively, this study suggests that the Cas13a-based multiplex RNA targeting system can be utilized to engineer resistance to RNA viruses in plants, whereas the number and expression levels of gRNAs have no significant effect on CRISPR/Cas13a-mediated viral interference in plants.
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Potyvirus , Potyvirus/genética , ARN , Sistemas CRISPR-Cas/genética , Plantas Modificadas Genéticamente/genética , Procesamiento Postranscripcional del ARNRESUMEN
OBJECTIVES: Early screening and dynamic monitoring of pulmonary vascular disease (PVD) in bronchopulmonary dysplasia (BPD) high-risk infants is of great clinical significance. Pulmonary artery acceleration time (PAAT) is a reliable and non-invasive method for assessing PVD in children over 1 year, but to date, few studies have used PAAT to assess pulmonary hemodynamics of preterm infants, especially those with BPD. Through dynamic monitoring the main hemodynamic indicators reflected PVD after birth, this study aimed to assess the value of PAAT in evaluating early PVD in BPD infants. METHODS: All 81 preterm infants at risk of BPD were divided into BPD and non-BPD groups according to whether BPD occurred. Clinical characteristics, PAAT, right ventricular ejection time (RVET) and other main hemodynamic indicators at four different time points after birth were studied and compared. RESULTS: PAAT and PAAT/RVET increased gradually within 72 h after birth in the BPD group (p < .05), but the curve tended to be flat over time after 72 h (p > .05). At PMA32 and 36 weeks, the PAAT (49.7 ± 4.8 vs. 54.8 ± 5.7, p = .001; 50.0 ± 5.3 vs. 57.0 ± 5.3, p = .001) and PAAT/RVET (.33 ± .04 vs. .35 ± .03, p = .001; .34 ± .03 vs. .37 ± .04, p = .001) in BPD group were significantly lower than those in the non-BPD group. CONCLUSIONS: PAAT and PAAT/RVET in the BPD group infants showed different change patterns compared to non-BPD group infants. PAAT can be used as a noninvasive and reliable screening method for screening and dynamic monitoring of PVD in BPD high-risk infants.
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Displasia Broncopulmonar , Enfermedades Vasculares , Lactante , Niño , Recién Nacido , Humanos , Recien Nacido Prematuro , Arteria Pulmonar , AceleraciónRESUMEN
An unsophisticated fluorescence-enabled strategy is brought forward to process the highly sensitive fluorescence detection of Salmonella typhimurium (S. typhimurium) which based on polyethyleneimine (PEI)-templated silver/copper nanoclusters (Ag/CuNCs) (λ excitation = 334 nm and λ emission = 466 nm) with cryonase-assisted target recycling amplification. The Ag/CuNCs nanoclusters are synthesized as fluorescent materials due to their strong and stable fluorescence characteristics and are modified with S. typhimurium aptamers to form aptamer-Ag/CuNCs probes. The probes can be adsorbed on the surface of quenching agents-polydopamine nanospheres (PDANSs), thereby inducing fluorescence quenching of the probes. Once the aptamers are bound to the target, the aptamers/targets complexes are separated from the PDANSs surface, and the Ag/CuNCs recover the fluorescence signal. The released complexes will immediately be transformed into a substrate digested by cryonase (an enzyme that can digest all types of nucleic acids), and the released targets are bound to another aptamers to initiate the next round of cleavage. This reaction will be repeated continuously until all relevant aptamers are consumed and all Ag/CuNCs are released, resulting in a significant amplification of the fluorescence signal and improved sensitivity. Using Ag/CuNCs as fluorescent probes combined with cryonase-assisted amplification strategy, the fluorescence aptasensor is constructed with detection limits as low as 3.8 CFU mL-1, which is tenfold better than without the cryonase assistance. The method developed has been applied to milk, orange juice, chicken, and egg white samples with excellent selectivity and accuracy providing an approach for the early and rapid detection of S. typhimurium in food.
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Cobre , Salmonella typhimurium , Animales , Plata , Pollos , Colorantes Fluorescentes , OligonucleótidosRESUMEN
The total viable count (TVC) of bacteria is an important index to evaluate the freshness and safety of dishes. To improve the accuracy and robustness of spectroscopic detection of total viable bacteria count in a complex system, a new method based on a near-infrared (NIR) hyperspectral hybrid model and Support Vector Machine (SVM) algorithms was developed to directly determine the total viable count in intact beef dish samples in this study. Diffuse reflectance data of intact and crushed samples were tested by NIR hyperspectral and processed using Multiplicative Scattering Correction (MSC) and Competitive Adaptive Reweighted Sampling (CARS). Kennard-Stone (KS) and Samples Set Partitioning Based on Joint X-Y Distance (SPXY) algorithms were used to select the optimal number of standard samples transferred by the model combined with root mean square error. The crushed samples were transferred into the complete samples prediction model through the Direct Standardization (DS) algorithm. The spectral hybrid model of crushed samples and full samples was established. The results showed that the Determination Coefficient of Calibration (RP2) value of the total samples prediction set increased from 0.5088 to 0.8068, and the value of the Root Mean Square Error of Prediction (RMSEP) decreased from 0.2454 to 0.1691 log10 CFU/g. After establishing the hybrid model, the RMSEP value decreased by 9.23% more than before, and the values of Relative Percent Deviation (RPD) and Reaction Error Relation (RER) increased by 12.12% and 10.09, respectively. The results of this study showed that TVC instewed beef samples can be non-destructively determined based on the DS model transfer method combined with the hybrid model strategy. This study provided a reference for solving the problem of poor accuracy and reliability of prediction models in heterogeneous samples.
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Algoritmos , Espectroscopía Infrarroja Corta , Animales , Bovinos , Espectroscopía Infrarroja Corta/métodos , Reproducibilidad de los Resultados , Análisis de los Mínimos Cuadrados , Máquina de Vectores de SoporteRESUMEN
This study aims to examine the changes in myocardial microcirculation in rats in a high-altitude hypoxic environment via computed tomography (CT) myocardial perfusion imaging technology. Rats in two groups were raised in different environments from 4 weeks of age for a period of 24 weeks. At 28 weeks of age, both groups underwent CT myocardial perfusion scanning, and the following myocardial perfusion parameters were measured: time to peak (TTP), mean transit time (MTT), blood flow (BF), and blood volume (BV). Following the scan, the rats were sacrificed, the cardiac index and right ventricular hypertrophy index were obtained, and hematoxylin-eosin (HE) staining was utilized to observe the pathological changes in the myocardium. In the group of rats that are subject to a high-altitude hypoxic environment for 24 weeks (the high-altitude group), the TTP and MTT values were increased (P < 0.05), the BF and BV values were lower (P < 0.05), the right heart mass was higher (P < 0.05) than that in the low-altitude group. As shown by the pathological results of HE staining, the gap between cardiomyocytes in the high-altitude group was widened, the arrangement of cardiomyocytes was irregular, and the cells were filled with a few fat vacuoles. The myocardial microcirculation is altered in a high-altitude hypoxic environment. In particular, the myocardium is in a state of inadequate perfusion, the BF in the myocardium slows down, and the right heart displays compensatory hypertrophy.
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Altitud , Imagen de Perfusión Miocárdica , Ratas , Animales , Microcirculación , Tomografía Computarizada por Rayos X/métodos , Hipoxia , Miocardio , Imagen de PerfusiónRESUMEN
Plant-parasitic nematodes cause huge agricultural economic losses. Two major families of Bacillus thuringiensis crystal proteins, Cry5 and Cry6, show nematicidal activity. Previous work showed that binding to midgut receptors is a limiting step in Cry toxin mode of action. In the case of Cry5Ba, certain Caenorhabditis elegans glycolipids were identified as receptors of this toxin. However, the receptors for Cry6 toxin remain unknown. In this study, the C. elegans CUB-like-domain containing protein RBT-1, released by phosphatidylinositol-specific phospholipase C (PI-PLC), was identified as a Cry6Aa binding protein by affinity chromatography. RBT-1 contained a predicted glycosylphosphatidylinositol (GPI) anchor site and was shown to locate in lipid rafts in the surface of the midgut cells. Western ligand blot assays and ELISA binding analysis confirmed the binding interaction between Cry6Aa and RBT-1 showing high affinity and specificity. In addition, the mutation of rbt-1 gene decreased the susceptibility of C. elegans to Cry6Aa but not that of Cry5Ba. Furthermore, RBT-1 mediated the uptake of Cry6Aa into C. elegans gut cells, and was shown to be involved in triggering pore-formation activity, indicating that RBT-1 is required for the interaction of Cry6Aa with the nematode midgut cells. These results support that RBT-1 is a functional receptor for Cry6Aa.
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Bacillus thuringiensis/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Endotoxinas/metabolismo , Proteínas Hemolisinas/metabolismo , Receptores de Superficie Celular/metabolismo , Animales , Toxinas de Bacillus thuringiensis , Caenorhabditis elegans/genética , Caenorhabditis elegans/microbiología , Proteínas de Caenorhabditis elegans/genética , Microdominios de Membrana/genética , Microdominios de Membrana/metabolismo , Mutación , Oligosacáridos/genética , Oligosacáridos/metabolismo , Receptores de Superficie Celular/genéticaRESUMEN
Allantoin is an important fine chemical that can be widely used in pharmaceutical, cosmetic and agricultural industries. Currently, allantoin is mainly produced by plant extraction or chemical synthesis. Due to the cost and environmental concerns, biosynthesis of allantoin from renewable feedstock is much more desirable. However, microbial production of allantoin from simple carbon sources has not yet been achieved so far. In this study, de novo biosynthesis of allantoin was achieved by constructing an artificial biosynthetic pathway. First, screening of efficient urate oxidases and xanthine dehydrogenases enabled allantoin production from hypoxanthine, a natural intermediate in purine metabolic pathway in Escherichia coli. Then, assemble of the entire pathway resulted in 13.9 mg/L allantoin from glucose in shake flask experiments. The titer was further improved to 639.8 mg/L by enhancing the supply of the precursor, redistribution of carbon flux, and reduction of acetate. Finally, scale-up production of allantoin was conducted in a 1-L fermentor under fed-batch culture conditions, which enabled the synthesis of 2360 mg/L allantoin, representing a 170-fold increase compared with the initial strain. This study not only demonstrates the potential for industrial production of allantoin, but also provides a bacterial platform for synthesis of other purines-derived high-value chemicals.
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Alantoína , Escherichia coli , Alantoína/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Ingeniería Metabólica/métodos , Redes y Vías Metabólicas , Urato Oxidasa/genética , Urato Oxidasa/metabolismoRESUMEN
OBJECTIVE: This study aimed to investigate the diagnostic value of doing a second ultrasound-guided fine-needle aspiration (US-FNA) for thyroid nodules of different sizes that could not be diagnosed by the first US-FNA. METHODS: One hundred and forty-three patients (162 nodules) were diagnosed with suspected malignant thyroid nodules in a routine ultrasound examination, but since the diagnosis could not be confirmed by the cytology of the samples collected in the first US-FNA, the patients underwent US-FNA again 3 months later. The ultrasound results, cytology results, and postoperative pathology of these nodules were collected. The nodules were divided into three groups according to the largest diameter (L) of the thyroid nodules: Group 1, L < 0.5 cm, 26 nodules; Group 2, L = 0.5-1.0 cm, 76 nodules; and Group 3, L > 1.0 cm, 60 nodules. RESULTS: In the second US-FNA, the overall diagnosis rate of the 162 thyroid nodules that could not be given a definitive diagnosis by the first US-FNA was 51.8% (84/162). The definitive diagnosis rates of the nodules in Groups 1, 2, and 3 were 30.8% (8/26), 67.1% (51/76), and 41.7% (25/60), respectively. The diagnosis rate was the highest in Group 2, and the differences between this group and the other two groups were statistically significant (χ2 = 10.489, 8.801, p < 0.05 for both). The diagnostic accuracy rates of Groups 1, 2, and 3 were 100% (8/8), 96.1% (49/51), and 92% (23/25), respectively. CONCLUSION: Second US-FNA is highly recommended for such nodules.
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Neoplasias de la Tiroides , Nódulo Tiroideo , Biopsia con Aguja Fina , Humanos , Biopsia Guiada por Imagen , Estudios Retrospectivos , Neoplasias de la Tiroides/patología , Nódulo Tiroideo/diagnóstico por imagen , Nódulo Tiroideo/patología , Ultrasonografía Intervencional/métodosRESUMEN
Pore-forming toxins (PFTs) are important weapons of multiple bacterial pathogens to establish their infections. PFTs generally form pores in the plasma membrane of target cells; however, the intracellular pathogenic processes triggered after pore-formation remain poorly understood. Using Caenorhabditis elegans as a model and Bacillus thuringiensis nematicidal Cry PFTs, we show here that the localized PFT attack causes a systemic mitochondrial damage, important for the PFT toxicity. We find that PFTs punch pores only in gut cells of nematodes, but unexpectedly mitochondrial disruption is able to occur in distal unperforated regions, such as the head and muscle tissues. We demonstrate that PFTs affect the activity of the mitochondrial respiratory chain (MRC) complex I resulting in the loss of mitochondrial membrane potential (ΔΨm ), which causes further mitochondrial fragmentation and the reduction of total mitochondrial content. Worms with decreased ΔΨm or inhibited MRC activity show higher sensitivity to PFTs. The inhibition of mitochondrial fission or the increase of mitochondrial content markedly improves the survival of animals treated with PFTs. These findings suggest that mitochondrial changes underpin PFT-mediated toxicity against nematodes and that systemic mitochondrial disruption caused by localized pore-formation represents a conserved key intracellular event in the mode of action of PFTs.
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Toxinas Bacterianas , Caenorhabditis elegans , Animales , Bacterias , Membrana CelularRESUMEN
Human astrovirus (HAstV) is one of the most common causative agents of acute gastroenteritis in children with an infection rate estimated to range from 2% to 9% worldwide. This study was aimed at investigating the molecular and clinical epidemiological features of human astrovirus infections in children under 5 years old with acute gastroenteritis in Shandong province, China from July 2017 to June 2018. In total, 376 fecal samples and the corresponding clinical information were collected and analyzed. HAstV infections were detected in all age groups with an overall positive rate of 8.51%. In addition to acute diarrhea, the main clinical manifestations were fever, abdominal pain, vomiting, and dehydration, in which fever was the most common complication. Infections could be seen throughout the year with a peak in the colder season. Four genotypes were detected in which HAstV-1 was the most prevalent genotype with a prevalence of 78.12%, followed by HAstV-5 (9.38%), MLB-1 (9.38%), and MLB-2 (3.12%). HAstV-1 strains were classified as lineage 1a, 1b, and 1d, in which lineage 1a strains were the most prevalent followed by lineage 1b and lineage 1d strains. All HAstV-5 strains were classified as lineage 5b and no other lineages were detected. The results showed that HAstV infection was an important cause of acute gastroenteritis among children under 5 years old in Shandong province. Given that their disease spectrum had been broadened, HAstVs should be paid more attention, not only as a causative agent of acute gastroenteritis but also as a potential pathogen of unexpected diseases.
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Infecciones por Astroviridae/epidemiología , Infecciones por Astroviridae/virología , Gastroenteritis/epidemiología , Gastroenteritis/virología , Mamastrovirus/genética , Enfermedad Aguda , Infecciones por Astroviridae/diagnóstico , Preescolar , China/epidemiología , Heces/virología , Femenino , Gastroenteritis/diagnóstico , Genotipo , Humanos , Lactante , Recién Nacido , Masculino , Mamastrovirus/clasificación , Epidemiología Molecular , Filogenia , Prevalencia , ARN Viral/genética , Estaciones del AñoRESUMEN
Rotaviruses are important causative agents of acute gastroenteritis in children. In China, rotavirus infection has a prevalence rate of 30% and is therefore considered a serious public health problem. This study was carried out to investigate the clinical and molecular epidemiological characteristics of rotavirus infections in children under 5 years old with acute diarrhea in Shandong province, China. From July 2017 to June 2018, a total of 1211 fecal specimens were tested, and the prevalence of rotavirus infection was 32.12%. The mean age of the infected children was 12.2 ± 10.9 months, and the highest infection rate was observed in children aged 7-12 months, with a rate of 41.64%. G9P[8] (76.61%) was the most prevalent genotype combination, followed by G2P[4] (7.20%), G3P[8] (3.60%), and G9P[4] (2.06%). In addition to diarrhea, vomiting, fever, and dehydration were the most common clinical signs. In general, there was no significant difference in clinical manifestations among different age groups. However, the clinical manifestations differed significantly between vaccinated and unvaccinated children. Vaccinated children showed lower incidence and frequency of vomiting, lower incidence and degree of dehydration, and lower incidence of severe cases than unvaccinated children. These findings suggest that it is necessary to continuously monitor changes in the characteristics of rotavirus infections. Moreover, the introduction of vaccines into the national immunization program to prevent and control rotavirus infection is needed in China.
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Epidemiología Molecular , Infecciones por Rotavirus/epidemiología , Infecciones por Rotavirus/virología , Rotavirus/genética , Preescolar , China/epidemiología , Diarrea/virología , Femenino , Gastroenteritis/virología , Genotipo , Humanos , Programas de Inmunización , Incidencia , Lactante , Masculino , Prevalencia , Infecciones por Rotavirus/fisiopatología , Infecciones por Rotavirus/prevención & control , Vacunas contra Rotavirus , VacunaciónRESUMEN
MicroRNAs (miRNAs) are important posttranscriptional regulators of gene expression. However, comprehensive expression profiles of miRNAs during mammalian spermatogenesis are lacking. Herein, we sequenced small RNAs in highly purified mouse spermatogenic cells at different stages. We found that a family of X-linked miRNAs named spermatogenesis-related miRNAs (spermiRs) is predominantly expressed in the early meiotic phases and has a conserved testis-specific high expression pattern in different mammals. We identified one spermiR homolog in opossum; this homolog might originate from THER1, a retrotransposon that is active in marsupials but extinct in current placental mammals. SpermiRs have expanded rapidly with mammalian evolution and are diverged into two clades, spermiR-L and spermiR-R, which are likely to have been generated at least in part by tandem duplication mediated by flanking retrotransposable elements. Notably, despite having undergone highly frequent lineage-specific duplication events, the sequences encoding all spermiR family members are strictly located between two protein-coding genes, Slitrk2 and Fmr1. Moreover, spermiR-Ls and spermiR-Rs have evolved different expression patterns during spermatogenesis in different mammals. Intriguingly, the seed sequences of spermiRs, which are critical for the recognition of target genes, are highly divergent within and among mammals, whereas spermiR target genes largely overlap. When miR-741, the most highly expressed spermiR, is knocked out in cultured mouse spermatogonial stem cells (SSCs), another spermiR, miR-465a-5p, is dramatically upregulated and becomes the most abundant miRNA. Notably, miR-741-/- SSCs grow normally, and the genome-wide expression levels of mRNAs remain unchanged. All these observations indicate functional compensation between spermiR family members and strong coevolution between spermiRs and their targets.
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Evolución Molecular , Genes Ligados a X , Mamíferos/genética , MicroARNs/genética , Espermatozoides/metabolismo , Animales , Secuencia de Bases , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/genética , Masculino , Mamíferos/metabolismo , Proteínas de la Membrana/genética , Ratones , Familia de Multigenes , Proteínas del Tejido Nervioso/genética , Testículo/metabolismoRESUMEN
Small noncoding RNAs (sncRNAs) regulate many genes in eukaryotic cells. Hua enhancer 1 (Hen1) is a 2'-O-methyltransferase that adds a methyl group to the 2'-OH of the 3'-terminal nucleotide of sncRNAs. The types and properties of sncRNAs may vary among different species, and the domain composition, structure, and function of Hen1 proteins differ accordingly. In mammals, Hen1 specifically methylates sncRNAs called P-element-induced wimpy testis-interacting RNAs (piRNAs). However, other types of sncRNAs that are methylated by Hen1 have not yet been reported, and the structures and the substrates of mammalian Hen1 remain unknown. Here, we report that mouse Hen1 (mHen1) performs 3'-end methylation of classical piRNAs, as well as those of most noncanonical piRNAs derived from rRNAs, small nuclear RNAs and tRNAs in murine spermatogonial stem cells. Moreover, we found that a distinct class of tRNA-derived sncRNAs are mHen1 substrates. We further determined the crystal structure of the putative methyltransferase domain of human Hen1 (HsHen1) in complex with its cofactor AdoMet at 2.0 Å resolution. We observed that HsHen1 has an active site similar to that of plant Hen1. We further found that the putative catalytic domain of HsHen1 alone exhibits no activity. However, an FXPP motif at its N terminus conferred full activity to this domain, and additional binding assays suggested that the FXPP motif is important for substrate binding. Our findings shed light on its methylation substrates in mouse spermatogonial stem cells and the substrate-recognition mechanism of mammalian Hen1.
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Dominio Catalítico , Metiltransferasas/química , Metiltransferasas/metabolismo , Espermatogonias/citología , Células Madre/metabolismo , Secuencias de Aminoácidos , Animales , Secuencia Conservada , Humanos , Masculino , Metilación , Ratones , Modelos Moleculares , Unión Proteica , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , ARN Nuclear Pequeño/genética , ARN Nuclear Pequeño/metabolismo , Especificidad por SustratoRESUMEN
CRISPR/Cas systems provide bacteria and archaea with molecular immunity against invading phages and foreign plasmids. The class 2 type VI CRISPR/Cas effector Cas13a is an RNA-targeting CRISPR effector that provides protection against RNA phages. Here we report the repurposing of CRISPR/Cas13a to protect potato plants from a eukaryotic virus, Potato virus Y (PVY). Transgenic potato lines expressing Cas13a/sgRNA (small guide RNA) constructs showed suppressed PVY accumulation and disease symptoms. The levels of viral resistance correlated with the expression levels of the Cas13a/sgRNA construct in the plants. Our data further demonstrate that appropriately designed sgRNAs can specifically interfere with multiple PVY strains, while having no effect on unrelated viruses such as PVA or Potato virus S. Our findings provide a novel and highly efficient strategy for engineering crops with resistances to viral diseases.
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Sistemas CRISPR-Cas , Resistencia a la Enfermedad/genética , Enfermedades de las Plantas/genética , Potyvirus/patogenicidad , Solanum tuberosum/genética , Enfermedades de las Plantas/virología , ARN Guía de Kinetoplastida/genética , Solanum tuberosum/virologíaRESUMEN
In this paper, we report the synthesis, the structural and optical characterization of CdSe/CdS//CdS nanorods (NRs) and their exploitation in nanorod-based light-emitting diodes (NR-LEDs). Two kinds of NRs of CdSe/CdS and CdSe/CdS//CdS were incorporated into the structure of solution-processed hybrid NR-LEDs. Compared to CdSe/CdS, the efficiencies of CdSe/CdS//CdS NR-based LEDs are overwhelmingly higher, specifically showing unprecedented values of peak current efficiency of 19.8 cd/A and external quantum efficiency of 15.7%. Such excellent results are likely attributable to a unique structure in CdSe/CdS//CdS NRs with a relatively high quantum yield, thick CdS outer shell, and rod structure which minimize nonradiative energy transfer between closely packed NRs in emitting layer.
RESUMEN
Cell death plays an important role in host-pathogen interactions. Crystal proteins (toxins) are essential components of Bacillus thuringiensis (Bt) biological pesticides because of their specific toxicity against insects and nematodes. However, the mode of action by which crystal toxins to induce cell death is not completely understood. Here we show that crystal toxin triggers cell death by necrosis signaling pathway using crystal toxin Cry6Aa-Caenorhabditis elegans toxin-host interaction system, which involves an increase in concentrations of cytoplasmic calcium, lysosomal lyses, uptake of propidium iodide, and burst of death fluorescence. We find that a deficiency in the necrosis pathway confers tolerance to Cry6Aa toxin. Intriguingly, the necrosis pathway is specifically triggered by Cry6Aa, not by Cry5Ba, whose amino acid sequence is different from that of Cry6Aa. Furthermore, Cry6Aa-induced necrosis pathway requires aspartic protease (ASP-1). In addition, ASP-1 protects Cry6Aa from over-degradation in C. elegans. This is the first demonstration that deficiency in necrosis pathway confers tolerance to Bt crystal protein, and that Cry6A triggers necrosis represents a newly added necrosis paradigm in the C. elegans. Understanding this model could lead to new strategies for nematode control.
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Proteasas de Ácido Aspártico/metabolismo , Proteínas Bacterianas/metabolismo , Caenorhabditis elegans/enzimología , Endotoxinas/metabolismo , Proteínas Hemolisinas/metabolismo , Secuencia de Aminoácidos , Animales , Proteasas de Ácido Aspártico/química , Toxinas de Bacillus thuringiensis , Calorimetría , Electroforesis en Gel Bidimensional , Ensayo de Inmunoadsorción Enzimática , Interacciones Huésped-Patógeno , Immunoblotting , Espectrometría de Masas , Datos de Secuencia Molecular , Necrosis , Control Biológico de Vectores/métodosRESUMEN
Invasive species often cause enormous economic and ecological damage, and this is especially true for invasive plants in the Asteraceae family. Arbuscular mycorrhizal fungi (AMF) play an important role in the successful invasion by exotic plant species because of their ability to promote growth and influence interspecific competition. However, few studies have evaluated the effects of invasive Asteraceae species on AMF diversity and how feedback mechanisms during competition with native species subsequently affect the accumulation of nutrient resources. Two exotic Asteraceae, Ambrosia artemisiifolia and Bidens pilosa, were monitored during competition with a native grass species, Setaria viridis, which is being replaced by these exotic species in natural areas around the study site. From these species continuously maintained in a field plot for 5 years, we collected the rhizosphere soil and cloned and identified soil AMF. Furthermore, AM fungal spores were isolated from rhizosphere soil of the two invasive species and used as inoculum in greenhouse experiments, to compare growth and nutrient accumulation during competition. The results indicate that although the AMF diversity in the rhizosphere soil of A. artemisiifolia and B. pilosa differed, the three most abundant species (Septoglomus viscosum, Septoglomus constrictum, Glomus perpusillum) were identical. The addition of AMF inoculum changed the competition between the plants, increasing the competitive ability of the invasives and decreasing that of the native. The results show a similar AMF community composition between A. artemisiifolia and B. pilosa, increased AMF root colonization of the invasive species during competition, AMF-enhanced N accumulation, and AMF-facilitated competitive growth of the invasive species.
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Ambrosia/crecimiento & desarrollo , Bidens/crecimiento & desarrollo , Glomeromycota/fisiología , Micorrizas/fisiología , Setaria (Planta)/crecimiento & desarrollo , Microbiología del Suelo , Ambrosia/microbiología , Bidens/microbiología , China , Especies IntroducidasRESUMEN
OBJECTIVES: This study aimed to evaluate the diagnostic value of acoustic radiation force impulse (ARFI) imaging and strain elastography in differentiating malignant from benign thyroid nodules. METHODS: This study included 128 patients (104 female and 24 male; mean age ± standard deviation, 48 ± 11 years; range, 23-76 years) with 152 pathologically proven thyroid nodules. Conventional sonography, strain ratio (SR) of strain elastography, virtual touch tissue imaging (VTI) grade, and virtual touch tissue quantification (shear wave velocity [SWV]) from ARFI imaging data were generated for each nodule. The diagnostic performance of the three elastography analysis methods was analyzed and compared by multiple receiver operating characteristic curve analysis. The interobserver agreement for VTI grade was also assessed. RESULTS: The best cutoff values for VTI grade, mean SWV, and mean SR for predicting malignant thyroid nodules were greater than or equal to grade χ, 2.87 m/s and 2.37, respectively. The area under the receiver operating characteristic curve for VTI grade, mean SWV, and mean SR was 0.854, 0.869 and 0.849, respectively (P > .05), and the accuracy was 83.55, 81.58 and 80.26%, respectively (P > .05). The accuracy of the combined use of conventional sonography and ARFI imaging for VTI grade, mean SWV, and mean SR was 98.03, 95.39 and 96.71%, respectively, which was slightly higher than that of conventional sonography (P > .05). The interobserver agreement produced a κ-value of 0.98 (95% confidence interval, 0.959-1.000). CONCLUSIONS: Both strain elastography and ARFI imaging have high sensitivity and specificity for differentiating malignant from benign thyroid nodules. Combined with sonography, these techniques can improve the accuracy of thyroid nodule diagnosis.
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Diagnóstico por Imagen de Elasticidad/métodos , Glándula Tiroides/diagnóstico por imagen , Neoplasias de la Tiroides/diagnóstico por imagen , Nódulo Tiroideo/diagnóstico por imagen , Adulto , Anciano , Diagnóstico Diferencial , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Pyruvate kinase M2 (PKM2) plays a pivotal role in the growth, survival and metabolic reprogramming of cancer cells. Here, we presented for the first time that tanshinone â ¡A inhibited human esophagus cancer cell growth through miR-122-mediated PKM2 down-regulation pathway. Tanshinone â ¡A inhibited cell proliferation and induced cell cycle arrest in S phase in human Ec109 cells. As expected, tanshinone â ¡A down-regulated PKM2 mRNA and protein expression in Ec109 cells. Given these findings, we further investigated microRNAs regulation of PKM2 and confirmed miR-122 for targeting PKM2. Moreover, we found that tanshinone â ¡A-induced up-regulation of miR-122 expression inhibited PKM2 expression in Ec109 cells. Meanwhile, tanshinone â ¡A inhibited proliferation through miR122-medated PKM2 down-regulation. It was demonstrated that the anticancer activity of tanshinone â ¡A was targeted at metabolic regulation of miR-122/PKM2 in human esophagus cancer cells. Taken together, our results revealed tanshinone â ¡A targeting at PKM2-mediated metabolic reprogramming play an important role in inhibition of esophageal cancer cell growth.
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Abietanos/farmacología , Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Neoplasias Esofágicas/tratamiento farmacológico , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , MicroARNs/biosíntesis , Proteínas de Neoplasias/biosíntesis , Piruvato Quinasa/biosíntesis , ARN Neoplásico/biosíntesis , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/patología , Células HeLa , HumanosRESUMEN
It has been reported that the trimethylation of histone 3 on lysine 27 (H3K27me3) is required for enhancer of zeste homology 2 (EZH2)-mediated repression of various genes essential for tumorigenesis and tumor development. Here, we reported the expression of EZH2 and H3K27me3 in esophageal squamous cell carcinoma (ESCC) specimens was higher than the pericarcinoma esophageal specimens. Their expression was positively associated with the poor prognosis of ESCC patients. EZH2 expression, histological grade and distant lymph node metastasis were all independent factors for poor prognosis of ESCC. In addition, enforced expression of EZH2 in esophageal cancer-derived cells could increase the overall H3K27me3 level. Our results suggested the expression of EZH2 and H3K27me3 could serve as biomarkers in the prediction of ESCC patients' survival and ESCC metastasis.