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BACKGROUND: Peanut is one of the most important oil and protein crops, and it exhibits wide cultivar variations in shoot Cd accumulation ability. However, the mechanism of Cd accumulation in peanut shoots has not been well understood. In this study, the root proteomics of two cultivars differing in seed Cd accumulation, Fenghua 1 (F, low Cd cultivar) and Silihong (S, high Cd cultivar), were investigated under 0 (CK) and 2 µM Cd conditions. RESULTS: A total of 4676 proteins were identified by proteomics screening. Of them, 375, 1762, 1276 and 771 proteins were identified to be differentially expressed proteins (DEPs) for comparison of FCd/FCK, SCd/SCK, FCK/SCK and FCd/SCd, respectively. Silihong is more sensitive to Cd exposure than Fenghua 1 in terms of root proteomics. A total of 30 and 86 DEPs were identified to be related with heavy metal transport and cell wall modification, respectively. The up-regulation of ABCB25, ABCC14, ABCC2, PDR1 and V-ATPases by Cd exposure in Silihong might enhance vacuolar sequestration of Cd and its efflux from symplast to apoplast. The higher Cd accumulation in the root CWs of Silihong might be resulted from its higher capability of CW modification, in which many proteins such as IRX10L, BGLU12-like, BGLU42, EXLB1, XTH30, XTH6, XYL7, PAL3, COMT, CAD1, and CCR1 were involved. CONCLUSIONS: The vacuolar sequestration and efflux of Cd as well as its adsorption in CW might be the principal mechanism of cadmium detoxification in Silihong. The higher capacity of Cd accumulation and translocation of Silihong is an inherent characteristics in which ACA8 and ZIP1 might be involved.
Assuntos
Arachis/metabolismo , Cádmio/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Proteômica , Transporte Biológico , Cádmio/toxicidade , Sementes/metabolismo , Especificidade da Espécie , Vacúolos/metabolismoRESUMO
The roles of virus-derived small RNAs (vsRNAs) have been studied in plants and insects. However, the generation and function of small RNAs from cytoplasmic RNA viruses in mammalian cells remain unexplored. This study describes four vsRNAs that were detected in enterovirus 71-infected cells using next-generation sequencing and northern blots. Viral infection produced substantial levels (>10(5) copy numbers per cell) of vsRNA1, one of the four vsRNAs. We also demonstrated that Dicer is involved in vsRNA1 generation in infected cells. vsRNA1 overexpression inhibited viral translation and internal ribosomal entry site (IRES) activity in infected cells. Conversely, blocking vsRNA1 enhanced viral yield and viral protein synthesis. We also present evidence that vsRNA1 targets stem-loop II of the viral 5' untranslated region and inhibits the activity of the IRES through this sequence-specific targeting. Our study demonstrates the ability of a cytoplasmic RNA virus to generate functional vsRNA in mammalian cells. In addition, we also demonstrate a potential novel mechanism for a positive-stranded RNA virus to regulate viral translation: generating a vsRNA that targets the IRES.
Assuntos
Regiões 5' não Traduzidas , Enterovirus Humano A/genética , Regulação Viral da Expressão Gênica , Biossíntese de Proteínas , Pequeno RNA não Traduzido/metabolismo , RNA Viral/metabolismo , Linhagem Celular Tumoral , Regulação para Baixo , Humanos , Ribonuclease III/metabolismo , Proteínas Virais/biossínteseRESUMO
Far-upstream element-binding protein 2 (FBP2) is an internal ribosomal entry site (IRES) trans-acting factor (ITAF) that negatively regulates enterovirus 71 (EV71) translation. This study shows that EV71 infection cleaved FBP2. Live EV71 and the EV71 replicon (but not UV-inactivated virus particles) induced FBP2 cleavage, suggesting that viral replication results in FBP2 cleavage. The results also showed that virus-induced proteasome, autophagy, and caspase activity co-contribute to EV71-induced FBP2 cleavage. Using FLAG-fused FBP2, we mapped the potential cleavage fragments of FBP2 in infected cells. We also found that FBP2 altered its function when its carboxyl terminus was cleaved. This study presents a mechanism for virus-induced cellular events to cleave a negative regulator for viral IRES-driven translation.
Assuntos
Enterovirus Humano A/metabolismo , Infecções por Enterovirus/metabolismo , Biossíntese de Proteínas/fisiologia , Proteínas de Ligação a RNA/metabolismo , Transativadores/metabolismo , Internalização do Vírus , Western Blotting , Linhagem Celular Tumoral , Humanos , Plasmídeos/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , RNA Interferente Pequeno/genética , Replicação Viral/fisiologiaRESUMO
PURPOSE: To investigate the efficacy of plasmin and sulfur hexafluoride (SF(6)) on the vitreoretinal junction, as well as the long-term safety in the eye and effect on the recipient's general health after application in the eye. METHODS: The study design included four groups of rabbits with three animals in each group. Group 1 received an intravitreal injection (IVI) of plasmin and SF(6) in the right eye; group 2 received an IVI of plasmin in the right eye; group 3 received an IVI of SF(6) in the right eye; and group 4 received an IVI of balanced salt solution in the right eye, which served as a normal control. Long-term safety (up to approximately three months) after plasmin and/or SF(6) injection was evaluated morphologically by clinical examination, histology, and immunohistochemistry, and functionally by electroretinograms (ERGs). General health evaluations after intravitreal injection included the assessment of weight gain, food intake, body temperature, and complete blood count analysis. RESULTS: Plasmin plus SF(6) injection resulted in complete posterior vitreous detachment (PVD), whereas plasmin or SF(6) injection alone resulted in only partial PVD. Balanced salt solution did not induce PVD. Eighty days after intravitreal injection, there were no major differences among the eyes of the three groups of animals compared with the normal control animals upon clinical evaluation, or regarding retinal morphology and ERGs. The lenses examined remained clear for up to 80 days following the intravitreal injection of plasmin plus SF(6), except one eye in the plasmin-treated group. ERGs decreased transiently one week after intravitreal injection in groups 1 through 3, but animals recovered fully to normal status afterward. General health was not affected after the injection of plasmin plus SF(6). CONCLUSIONS: Efficient vitreoretinal separation could be achieved, and an acceptable long-term safety profile was noted after plasmin plus SF(6) injection in the eye. No major ocular toxicity or systemic toxicity was found after the injection of plasmin plus SF(6). These results provide good support for the future clinical use of plasmin plus SF(6) for treatment of a variety of vitreoretinopathies.
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Fibrinolisina/administração & dosagem , Cristalino/efeitos dos fármacos , Retina/efeitos dos fármacos , Hexafluoreto de Enxofre/administração & dosagem , Corpo Vítreo/efeitos dos fármacos , Descolamento do Vítreo/induzido quimicamente , Animais , Combinação de Medicamentos , Eletrorretinografia , Imuno-Histoquímica , Injeções Intravítreas , Cristalino/citologia , Microscopia Eletrônica de Varredura , Proteólise/efeitos dos fármacos , Coelhos , Retina/citologia , Corpo Vítreo/citologiaRESUMO
Currently, clinical indications for the application of gadolinium-based contrast agents (GBCA) in magnetic resonance imaging (MRI) are increasingly being questioned. Consequently, this study aimed to evaluate the additional diagnostic value of contrast enhancement in MRI of the hand in patients with rheumatoid arthritis (RA). Thirty-one patients with RA (mean age, 50 ± 14 years (range, 18-72 years)) underwent morphologic MRI scans on a clinical 3 T scanner. MRI studies were analyzed based on (1) the Rheumatoid Arthritis Magnetic Resonance Imaging Score (RAMRIS) and (2) the GBCA-free RAMRIS version, termed RAMRIS Sine-Gadolinium-For-Experts (RAMRIS-SAFE), in which synovitis and tenosynovitis were assessed using the short-tau inversion-recovery sequence instead of the post-contrast T1-weighted sequence. The synovitis subscores in terms of Spearman's ρ, as based on RAMRIS and RAMRIS-SAFE, were almost perfect (ρ = 0.937; p < 0.001), while the tenosynovitis subscores were less strongly correlated (ρ = 0.380 p = 0.035). Correlation between the total RAMRIS and RAMRIS-SAFE was also almost perfect (ρ = 0.976; p < 0.001). Inter-rater reliability in terms of Cohen's κ was high (0.963 ≤ κ ≤ 0.925). In conclusion, RAMRIS-SAFE as the GBCA-free version of the well-established RAMRIS is a patient-friendly and resource-efficient alternative for assessing disease-related joint changes in RA. As patients with RA are subject to repetitive GBCA applications, non-contrast imaging protocols should be considered.
RESUMO
The development and characterization of an enhanced composite skin substitute based on collagen and poly(epsilon-caprolactone) are reported. Considering the features of excellent biocompatibility, easy-manipulated property and exempt from cross-linking related toxicity observed in the 1:20 biocomposites, skin substitutes were developed by seeding human single-donor keratinocytes and fibroblasts alone on both sides of the 1:20 biocomposite to allow for separation of two cell types and preserving cell signals transmission via micro-pores with a porosity of 28.8 +/- 16.1 microm. The bi-layered skin substitute exhibited both differentiated epidermis and fibrous dermis in vitro. Less Keratinocyte Growth Factor production was measured in the co-cultured skin model compared to fibroblast alone condition indicating a favorable microenvironment for epidermal homeostasis. Moreover, fast wound closure, epidermal differentiation, and abundant dermal collagen deposition were observed in composite skin in vivo. In summary, the beneficial characteristics of the new skin substitutes exploited the potential for pharmaceutical screening and clinical application.
Assuntos
Colágeno/química , Poliésteres/química , Pele Artificial , Animais , Técnicas de Cocultura , Fator 7 de Crescimento de Fibroblastos/biossíntese , Fibroblastos/metabolismo , Fibroblastos/fisiologia , Humanos , Queratinócitos/metabolismo , Queratinócitos/fisiologia , Teste de Materiais , Camundongos , Camundongos Nus , Porosidade , Doadores de TecidosRESUMO
Picornaviruses cause several diseases, not only in humans but also in various animal hosts. For instance, human enteroviruses can cause hand-foot-and-mouth disease, herpangina, myocarditis, acute flaccid paralysis, acute hemorrhagic conjunctivitis, severe neurological complications, including brainstem encephalitis, meningitis and poliomyelitis, and even death. The interaction between the virus and the host is important for viral replication, virulence and pathogenicity. This article reviews studies of the functions of viral and host factors that are involved in the life cycle of picornavirus. The interactions of viral capsid proteins with host cell receptors is discussed first, and the mechanisms by which the viral and host cell factors are involved in viral replication, viral translation and the switch from translation to RNA replication are then addressed. Understanding how cellular proteins interact with viral RNA or viral proteins, as well as the roles of each in viral infection, will provide insights for the design of novel antiviral agents based on these interactions.
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Infecções por Picornaviridae/virologia , Picornaviridae/fisiologia , Animais , Capsídeo/química , Genoma Viral , Humanos , Modelos Biológicos , Picornaviridae/genética , Infecções por Picornaviridae/metabolismo , Biossíntese de Proteínas , Interferência de RNA , RNA Viral/química , Proteínas Virais/química , Replicação ViralRESUMO
OBJECTIVE: In 2005, the Taiwan Institute of Occupational Safety and Health started an on-site consulting program for the medical supplies manufacturing industry, which use ethylene oxide (EO) as a sterilant, with the goal of enhancing occupational hygiene practices and controlling EO-related risks. This study presents EO exposure assessment results and examines the effectiveness of control measures. METHODS: Detailed surveys, including exposure monitoring, were conducted at 10 factories. Airborne EO was collected using an HBr-coated charcoal tube and analyzed using GC/MS. RESULTS: Sterilizer operators had an average short-term EO exposure level of 27.61 ppm during unloading; mean time-weighted average workshift exposure was 7.35 ppm. High EO concentrations were also present throughout the facilities. Specifically, mean EO concentrations in the aeration area, near the sterilizer and in the warehouse were 10.19, 5.75, and 8.78 ppm, respectively. These findings indicate that immediate controls are needed, and that EO emissions from sterilized products during storage cannot be overlooked. Worker short-term exposures during unloading was inversely correlated (p < 0.05) with the numbers of poststerilization purge cycle applied. The specific controls implemented and their usefulness is discussed. CONCLUSION: Increasing the number of poststerilization purge cycles is a simple approach to eliminating extremely high exposure during unloading. Improvements to ventilation, particularly in the aeration area and warehouse, were also effective in minimizing worker exposures. Use of effective respirator is recommended until the EO exposure levels, averaging 3.41 ppm after the controls, fall below the permissible exposure limit.
Assuntos
Poluentes Ocupacionais do Ar/análise , Óxido de Etileno/análise , Exposição por Inalação/prevenção & controle , Exposição Ocupacional/prevenção & controle , Esterilização , Poluentes Ocupacionais do Ar/toxicidade , Monitoramento Ambiental , Óxido de Etileno/toxicidade , Humanos , Indústrias , TaiwanRESUMO
PURPOSE: To investigate the rate of lens subluxation following plasmin and/or SF6 injections in eyes, and whether a subsequent elevated level of vascular endothelial growth factor (VEGF) and vitreous tap would aggravate subluxation. METHODS: Four groups of rabbits were used. Group 1 received an intravitreal injection (IVI) of plasmin and SF6 in the right eye; group 2 received an IVI of plasmin in the right eye; group 3 received an IVI of SF6 in the right eye; and group 4 received an IVI of balanced salt solution in the right eye. After treatment, IVIs of VEGF were given and vitreous tap was performed three times, followed by clinical observation of lens subluxation and scanning electronic microscope evaluation of the zonular fibers. RESULTS: After IVIs of plasmin and SF6, and VEGF and vitreous tap had been performed one to three times, lens subluxation was noted in 0%, 43%, 71%, 71%, and 86% of the eyes in group 1. After IVIs of plasmin, VEGF, and vitreous tap had been performed one to three times, lens subluxation was noted in 11%, 22%, 44%, 44%, and 67% of the eyes in group 2. The eyes in group 3 and 4 did not show signs of lens subluxation after VEGF IVIs and vitreous tap. Histology confirmed zonular fiber damage in the eyes treated with plasmin. CONCLUSIONS: The incidence of lens subluxation increased following plasmin injections in the eyes, and this was aggravated by the subsequent high VEGF level in the eyes and vitreous tapping. Zonular fibers were disrupted following plasmin treatment. These effects should be kept in mind when using plasmin enzymes in patients with vitreoretinal abnormalities.
Assuntos
Fibrinolisina/farmacologia , Subluxação do Cristalino/patologia , Cristalino/efeitos dos fármacos , Hexafluoreto de Enxofre/farmacologia , Animais , Injeções Intravítreas , Subluxação do Cristalino/metabolismo , Cristalino/patologia , Microscopia Eletrônica de Varredura , Coelhos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/patologia , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/farmacologiaRESUMO
In this paper, we numerically study an enhancement of breakdown voltage in AlGaN/GaN high-electron-mobility transistors (HEMTs) by using the AlGaN/GaN/AlGaN quantum-well (QW) electron-blocking layer (EBL) structure. This concept is based on the superior confinement of two-dimensional electron gases (2-DEGs) provided by the QW EBL, resulting in a significant improvement of breakdown voltage and a remarkable suppression of spilling electrons. The electron mobility of 2-DEG is hence enhanced as well. The dependence of thickness and composition of QW EBL on the device breakdown is also evaluated and discussed.
RESUMO
PURPOSE: To investigate the clearance of vascular endothelial growth factor (VEGF) after the induction of posterior vitreous detachment by plasmin and/or SF(6). METHODS: The study design included four groups of rabbits: group 1 received an intravitreal injection of plasmin and SF(6) in the right eye, group 2 received an intravitreal injection of plasmin in the right eye, group 3 received an intravitreal injection of SF(6) in the right eye, and group 4 received an intravitreal injection of balanced salt solution in the right eye. Intravitreal injection of human VEGF (50 µL, 10 ng/µL) was performed in study eyes and control eyes 1 month after plasmin and/or SF(6) injection. Serum and vitreous samples were collected on days 1, 3, and 7 after VEGF injection to determine the serum and vitreous concentrations of VEGF. RESULTS: One day after VEGF injection, residual human VEGF concentration in the vitreous cavity was significantly lower in the plasmin- and SF(6)-treated eyes (group 1) and the plasmin-treated eyes (group 2) when compared with the control eyes (group 4) (P = 0.047 and 0.027, respectively). Three days after VEGF injection, the residual VEGF concentration in the vitreous cavity was still significantly lower in the plasmin- and SF(6)-treated eyes (group 1) when compared with the control eyes (group 4) (P = 0.025). CONCLUSIONS: Eyes treated with plasmin exhibit a more rapid clearance of exogenous VEGF than control eyes. This finding suggests a novel treatment for retinopathies associated with vitreous traction and VEGF elevation.
Assuntos
Fibrinolisina/farmacologia , Hexafluoreto de Enxofre/farmacologia , Fator A de Crescimento do Endotélio Vascular/farmacocinética , Corpo Vítreo/metabolismo , Descolamento do Vítreo/metabolismo , Animais , Combinação de Medicamentos , Eletrorretinografia , Ensaio de Imunoadsorção Enzimática , Injeções Intravítreas , Microscopia Eletrônica de Varredura , Coelhos , Corpo Vítreo/efeitos dos fármacos , Descolamento do Vítreo/induzido quimicamenteRESUMO
Enterovirus 71 (EV71) is a neurotropic pathogen that can cause severe neural diseases and complications on infected patients. Clinical observations showed that EV71-induced immune responses may be associated with virus induced neurogenic pulmonary edema. Here reviewed studies that discovered several host molecules as potential factors for EV71 virulence.
Assuntos
Enterovirus Humano A/patogenicidade , Infecções por Enterovirus/virologia , Doenças do Sistema Nervoso/virologia , Apoptose , Infecções por Enterovirus/complicações , Interações Hospedeiro-Patógeno , Humanos , Sistema Nervoso/virologia , Sepse/complicações , Sepse/virologiaRESUMO
PURPOSE. To test the long-term effects and systemic exposure level after single or multiple bevacizumab (Avastin) intravitreal injections in newborn rabbit eyes. METHODS. Four groups of newborn New Zealand rabbits received a single intravitreal bevacizumab injection at a concentration of 1.25 mg/0.05 mL at the ages of 2 (group 1), 4 (group 2), 6 (group 3), and 12 (group 5) weeks. The other group of rabbits (group 4) received three consecutive injections of bevacizumab at a concentration of 1.25 mg/0.05 mL at weeks 2, 6, and 10. Eight days after injection, the serum concentration of bevacizumab was determined in groups 1, 2, 3, and 5. Morphologic and functional changes were evaluated 12 months after bevacizumab injection. RESULTS. Twelve months after either single or multiple intravitreal injections of bevacizumab, newborn rabbit eyes showed no significant differences compared with control eyes on examination with funduscopy, histopathology, or electroretinogram. The serum concentrations when the injections were performed at the ages of 2 (19.4 +/- 8.1 microg/mL) and 4 (10.2 +/- 2.3 microg/mL) weeks were significantly higher than the serum level detected when the injection was performed at 12 weeks of age (2.8 +/- 1.2 microg/mL, P = 0.02 and P = 0.024, respectively). CONCLUSIONS. After 1 year, single and three consecutive intravitreal injections of 1.25 mg bevacizumab in newborn rabbit eyes are well tolerated. Systemic exposure is higher when the injection is performed at an early age.
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Inibidores da Angiogênese/farmacocinética , Inibidores da Angiogênese/toxicidade , Anticorpos Monoclonais/farmacocinética , Anticorpos Monoclonais/toxicidade , Retina/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais Humanizados , Bevacizumab , Disponibilidade Biológica , Eletrorretinografia , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Proteína Glial Fibrilar Ácida/metabolismo , Injeções , Proteínas de Neurofilamentos/metabolismo , Opsinas/metabolismo , Coelhos , Retina/metabolismo , Retina/ultraestrutura , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Corpo VítreoRESUMO
Carbohydrate structures between retinal neurons and retinal pigment epithelium (RPE) play an important role in maintaining the integrity of retinal adhesion to underlying RPE, and in retinal detachment pathogenesis. Since relevant knowledge is still in the primary stage, glycotopes on the adult retina of mongrel canines (dog), micropigs and Sprague-Dawley rats were examined by lectino-histochemistry, using a panel of 16 different lectins. Paraffin sections of eyes were stained with biotinylated lectins, and visualized by streptavidin-peroxidase and diaminobenzidine staining. Mapping the affinity profiles, it is concluded that: (i) all sections of the retina reacted well with Morniga M, suggesting that N-linked glycans are present in all layers of the retina; (ii) no detectable human blood group ABH active glycotopes were found among retinal layers; (iii) outer and inner segments contained glycoconjugates rich in ligands reacting with T (alpha) (Galbeta1-3GalNAcalpha1-Ser/Thr) and Tn (GalNAcalpha1-Ser/Thr) specific lectins; (iv) cone cells of retina specifically bound peanut agglutinin (PNA), which recognizes T (alpha) residues and could be used as a specific marker for these photoreceptors; (v) the retinas of rat, dog and pig, had a similar binding profile but with different intensity; (vi) each retinal layer had its own binding characteristic. This information may provide useful background knowledge for normal retinal physiology and miscellaneous retinal diseases, including retinal detachment (RD) and age-related macular degeneration (ARMD).