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1.
Opt Express ; 31(1): 31-43, 2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36606947

RESUMO

We propose a new, to the best of our knowledge, method of incoherent optical frequency selection called three-pack frequency-selective incoherent holography. Compressed holography is reconstructed using phase shift intercepts and spatial transfer function convolution in the form of separation without loss of magnification or resolution. The frequency-selective reconstruction process removes the conjugate and DC terms along with the interception of the object wave. This work attempts three-dimensional reconstruction and selected-frequency phase extraction of axial slices in submicron steps, and the experimental results show the potential of the proposed method in areas such as compressed holography, extended field of view, and slice tomography.

2.
J Math Biol ; 86(5): 80, 2023 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-37093296

RESUMO

In this paper, we first formulate a system of ODEs-PDE to model diseases with latency-age and differential infectivity. Then, based on the ways how latent individuals leave the latent stage, one ODE and two DDE models are derived. We only focus on the global stability of the models. All the models have some similarities in the existence of equilibria. Each model has a threshold dynamics for global stability, which is completely characterized by the basic reproduction number. The approach is the Lyapunov direct method. We propose an idea on constructing Lyapunov functionals for the two DDE and the original ODEs-PDE models. During verifying the negative (semi-)definiteness of derivatives of the Lyapunov functionals along solutions, a novel positive definite function and a new inequality are used. The idea here is also helpful in applying the Lyapunov direct method to prove the global stability of some epidemic models with age structure or delays.


Assuntos
Epidemias , Modelos Biológicos , Humanos , Número Básico de Reprodução
3.
Angew Chem Int Ed Engl ; 61(38): e202210573, 2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-35909225

RESUMO

Sustainable processes for semi-hydrogenation of alkynes/alkadienes impurities in alkenes feedstocks are in great demand in industry as the utilization of excessive hydrogen, high temperature and unsatisfactory alkenes selectivity of the current thermo-catalytic route, however, their development is still challenging. Herein, we innovate a light-assisted semi-hydrogenation process in gas-feed fixed bed reactor, with water as hydrogen atom source by in situ photocatalysis. Using Pd/TiO2 as model catalyst, this process shows an excellent catalytic performance for the semi-hydrogenation of 1,3-butadiene, with 100 % of butenes selectivity at ≈99 % of conversion over 180 h of reaction at ambient temperature driven by 66 mW cm-2 of irradiation intensity. This light-driven, H2 -free, ambient temperature semi-hydrogenation process, with superior performance to that of thermocatalytic route, shows attractive to bring an evolution in industrial hydrogenation technology to an economical and safe way.

4.
Lab Invest ; 101(10): 1371-1381, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34239033

RESUMO

Sepsis is an acute inflammatory reaction and a cause of acute respiratory distress syndrome (ARDS). In the present study, we explored the roles and underlying mechanism of the lncRNA Nuclear enriched abundant transcript 1 (NEAT1) in ARDS. The expression levels of genes, proteins and pro-inflammatory cytokines in patients with ARDS, LPS-stimulated cells and septic mouse models were quantified using qPCR, western blotting and ELISA assays, respectively. The molecular targeting relationship was validated by conducting a dual-luciferase reporter assay. Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) assay. The cell cycle phase was determined by flow cytometry assay. The expression levels of NEAT1 and pro-inflammatory cytokines were higher in patients with ARDS and septic models than in controls. Knockdown of NEAT1 significantly increased cell proliferation and cycle progression and prolonged mouse survival in vitro and in vivo. Mechanistically, miR-27a was identified as a downstream target of NEAT1 and directly inhibited PTEN expression. Further rescue experiments revealed that inhibition of miR-27a impeded the promoting effects of NEAT1 silence on cell proliferation and cycle progression, whereas inhibition of PTEN markedly weakened the inhibitory effects of NEAT1 overexpression on cell proliferation and cycle progression. Altogether, our study revealed that NEAT1 plays a promoting role in the progression of ARDS via the NEAT1/miR-27a/PTEN regulatory network, providing new insight into the pathologic mechanism behind ARDS.


Assuntos
MicroRNAs/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , RNA Longo não Codificante , Síndrome do Desconforto Respiratório/metabolismo , Sepse/metabolismo , Adulto , Animais , Linhagem Celular , Modelos Animais de Doenças , Técnicas de Silenciamento de Genes , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/genética , PTEN Fosfo-Hidrolase/genética , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Transdução de Sinais/genética
5.
J Biol Chem ; 294(42): 15544-15556, 2019 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-31492755

RESUMO

The Escherichia coli cAMP receptor protein, CRP, is a homodimeric global transcription activator that employs multiple mechanisms to modulate the expression of hundreds of genes. These mechanisms require different interfacial interactions among CRP, RNA, and DNA of varying sequences. The involvement of such a multiplicity of interfaces requires a tight control to ensure the desired phenotype. CRP-dependent promoters can be grouped into three classes. For decades scientists in the field have been puzzled over the differences in mechanisms between class I and II promoters. Using a new crystal structure, IR spectroscopy, and computational analysis, we defined the energy landscapes of WT and 14 mutated CRPs to determine how a homodimeric protein can distinguish nonpalindromic DNA sequences and facilitate communication between residues located in three different activation regions (AR) in CRP that are ∼30 Šapart. We showed that each mutation imparts differential effects on stability among the subunits and domains in CRP. Consequently, the energetic landscapes of subunits and domains are different, and CRP is asymmetric. Hence, the same mutation can exert different effects on ARs in class I or II promoters. The effect of a mutation is transmitted through a network by long-distance communication not necessarily relying on physical contacts between adjacent residues. The mechanism is simply the sum of the consequences of modulating the synchrony of dynamic motions of residues at a distance, leading to differential effects on ARs in different subunits. The computational analysis is applicable to any system and potentially with predictive capability.


Assuntos
Proteína Receptora de AMP Cíclico/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Regiões Promotoras Genéticas , Sítios de Ligação , Proteína Receptora de AMP Cíclico/química , Proteína Receptora de AMP Cíclico/genética , Dimerização , Escherichia coli/química , Escherichia coli/genética , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Domínios Proteicos , Subunidades Proteicas/química , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo
6.
Opt Express ; 28(4): 5179-5188, 2020 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-32121743

RESUMO

In this paper, pure silk protein was extracted from Bombyx mori silks and fabricated into a new kind of disordered bio-microfiber structure using electrospinning technology. Coherent random lasing emission with low threshold was achieved in the silk fibroin fibers. The random lasing emission wavelength can be tuned in the range of 33 nm by controlling the pump location with different scattering strengths. Therefore, the bio-microfiber random lasers can be a wide spectral light source when the system is doped with a gain or energy transfer medium with a large fluorescence emission band. Application of the random lasers of the bio-microfibers as a low-coherence light source in speckle-free imaging had also been studied.


Assuntos
Fibroínas/química , Lasers , Luz , Animais , Bombyx , Fibroínas/ultraestrutura , Processamento de Imagem Assistida por Computador , Dispositivos Ópticos
7.
Sensors (Basel) ; 18(10)2018 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-30340312

RESUMO

A pulse-width multiplexing method for reducing the nuisance-alarm rate of a phase-sensitive optical time-domain reflectometer ( ϕ -OTDR) is described. In this method, light pulses of different pulse-widths are injected into the sensing fiber; the data acquired at different pulse-widths are regarded as the outputs of different sensors; and these data are then processed by a multisensor data fusion algorithm. In laboratory tests with a sensing fiber on a vibrating table, the effects of pulse-width on the signal-to-noise ratio (SNR) of the ϕ -OTDR data are observed. Furthermore, by utilizing the SNR as the feature in a feature-layer algorithm based on Dempster⁻Shafer evidential theory, a four-pulse-width multiplexing ϕ -OTDR system is constructed, and the nuisance-alarm rate is reduced by about 70%. These experimental results show that the proposed method has great potential for perimeter protection, since the nuisance-alarm rate is significantly reduced by using a simple configuration.

8.
J Theor Biol ; 424: 73-83, 2017 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-28479003

RESUMO

The epidemic characteristics of two classic SIS epidemic models, including the epidemic size, peak and turning point, are investigated. The two SIS models are with bilinear and standard incidences, respectively. For the SIS models, the susceptible individuals generally can be divided into two classes. One consists of the individuals who had not been infected by the infection, the other are individuals who have been infected and recovered from the infection. Based on this fact, the classic SIS epidemic models need to be reformulated in order to analyze the turning points of the epidemic for various cumulative cases in detail. The obtained results illustrate how to determine the epidemic characteristics of the two models, and demonstrate their dependence on the initial conditions and the relative parameters of the models.


Assuntos
Doenças Transmissíveis/mortalidade , Doenças Transmissíveis/transmissão , Infecções/mortalidade , Infecções/transmissão , Modelos Biológicos , Animais , Humanos
9.
Rapid Commun Mass Spectrom ; 31(17): 1424-1430, 2017 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-28586540

RESUMO

RATIONALE: Non-thermal plasma (NTP) has proven to be an effective approach for the removal of volatile organic compounds (VOCs). However, harmful organic by-products, produced during NTP-mediated removal of VOCs, hinder practical applications of this technology. It is necessary to characterize the organic by-products to assess their health risks. METHODS: A method is proposed for analyzing and evaluating organic by-products for NTP-mediated removal of VOCs in this work. NTP generated by a coaxial dielectric barrier discharge (DBD) reactor was used for the removal of a model VOC, toluene, in nitrogen. Organic products were characterized using a real-time proton transfer reaction time-of-flight mass spectrometry (PTR-TOF-MS) apparatus. RESULTS: The PTR-TOF-MS apparatus has been shown to be effective for real-time high-sensitivity detection of trace VOCs. The main observed organic compounds, with concentrations on the order of ppb/ppm, were hydrogen cyanide, acetonitrile, propanenitrile, benzene, benzonitrile, and benzyl nitrile, etc. CONCLUSIONS: A health-related index (HRI) was introduced to assess the health risks associated with these organic products. The HRI was not correlated with the removal efficiency (η), with higher η possibly yielding higher HRI, associated with higher health risks. Specific input energy (SIE) was a key factor affecting the formation of the observed organic products and their HRI values. We conclude that in practical applications, SIE, HRI, and η must be balanced.

10.
Phys Chem Chem Phys ; 19(33): 21938-21945, 2017 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-28612874

RESUMO

The self-assembly of a copolymer thin film, whose molecular structure is composed of one hydrophobic branch and two hydrophilic branches, gives a cylindrical structure consisting of a hydrophobic core and a hydrophilic outer surface. The confinement-induced loss of entropy leads the copolymer to self-assemble into a hexagonal arrangement of cylindrical structures. This hexagonal arrangement is of two induced self-assembly structures under one-dimensional confinement in thin films, i.e., micellar structures with uniform density of an individual core, as well as lamellar structures with two separated hydrophobic layers and one hydrophilic layer. When the repulsive force of the confinement is greater for the hydrophobic polymeric component than for the hydrophilic polymeric component, the self-assembled density of the cores is broad. A proportional change in the individual volume suggests interactions between the walls and the hydrophobic core, which plays a vital role in the formation of the self-assembled structure. A basic understanding of the one-dimensional confinement investigated in this study serves to elucidate the more complex two- and three-dimensional confinements and provides further insights for the design of nanomaterials with novel morphologies.

11.
Phys Chem Chem Phys ; 19(46): 31011-31023, 2017 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-28967926

RESUMO

Different from the self-assembly of neutral polymers, polyelectrolytes self-assemble into smaller aggregates with a more loosely assembled structure, which results from the repulsive forces acting between similar electrical compositions with the introduction of ions. The Y-shaped weak polyelectrolytes self-assemble into a core-shell type cylindrical structure with a hexagonal arrangement in a thin film, whose thickness is smaller than the gyration radius of the polymer chain. The corresponding formation mechanism consists of enrichment of the same components, adjustment of the shape of the aggregate, and the subsequent separation into individual aggregates. With the increase in the thickness of the thin film until it exceeds the gyration radius of the polymer chain, combined with the greater freedom of movement along the direction of thin film thickness, the self-assembled structure changes into a micellar structure. Under confinement, the repulsive force to the polymeric components is weakened by the repulsive forces among polyelectrolyte components with like charges, and this helps in generating aggregates with more uniform size and density distribution. In particular, when the repulsive force between the walls and the core forming components is greater than that between the walls and the shell forming components, such asymmetric confinement produces a crossed-cylindrical structure with nearly perpendicular arrangement of two cylinder arrays. Similarly, a novel three-crossed cylinder morphology is self-assembled upon removal of confinement.

12.
J Theor Biol ; 365: 337-46, 2015 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-25451526

RESUMO

A mathematical model for HCV infection is established, in which the effect of dendritic cells (DC) and cytotoxic T lymphocytes (CTL) on HCV infection is considered. The basic reproduction numbers of chronic HCV infection and immune control are found. The obtained results show that the infection dies out finally as the basic reproduction number of HCV infection is less than unity, and the infection becomes chronic as it is greater than unity. In the presence of chronic infection, the existence of immune control equilibrium is discussed completely, which illustrates that the backward bifurcation may occur under certain conditions, and that the two quantities, the sizes of the activated DC and the removed CTL during their average life-terms, play a critical role in controlling chronic HCV infection and immune response. The occurrence of backward bifurcation implies that there may be bistability for the model, i.e., the outcome of infection depends on the initial situation. By choosing the activated rate of non-activated DC or the cross-representation rate of activated DC as bifurcation number, Hopf bifurcation for certain condition shows the existence of periodic solution of the model. Again, numerical simulations suggest the dynamical complexity of the model including the instability of immune control equilibrium and the existence of stable periodic solution.


Assuntos
Hepacivirus/fisiologia , Hepatite C Crônica/imunologia , Hepatite C Crônica/virologia , Imunidade , Modelos Imunológicos , Simulação por Computador , Humanos , Análise Numérica Assistida por Computador
13.
Soft Matter ; 11(22): 4366-74, 2015 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-25871011

RESUMO

We designed twelve types of weak polyelectrolytes (i.e., PEO-b-PMMA copolymers (BCP) in multi-arm structures, where six include EO blocks as joint points and the other six have MMA blocks as joint points). All of the BCPs with EO as the joint points form disordered phases with the exception of long-chained and four-armed BCP. The main mesophases of all of the BCPs with MMA as joint points are micelle-like and bicontinuous phases. In particular, the short-chained BCP with four-arms and EO segments outside form a new phase type (i.e., crossed lamellar phase). Using MesoDyn, we provide a comprehensive representation of the micelle and crossed lamellar phase formation mechanisms based on both thermodynamic and dynamic analyses. A shear force on a micelle-like phase could promote a hexagonal columnar phase, which is a good technique for generating an ordered arrangement of nanotube arrays. Blending homopolymers with the same constituents could promote uniformity of the micelle size and decrease the polydispersity, especially for blends with a high BCP concentration, which may provide a new approach for regulating the properties of materials.


Assuntos
Modelos Moleculares , Polietilenoglicóis/química , Polimetil Metacrilato/química , Micelas
14.
Soft Matter ; 11(22): 4356-65, 2015 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-25875915

RESUMO

Combined with quantum calculations and mesoscale simulations, the self-assembly of twelve lipid-structured PEO-b-PMMA copolymers (BCPs) with six types of molecular topologies was investigated. The BCPs with MMA species as the connecting center of the other arms present ample mesoscale structures, such as micelles and lamellae or curved lamellar phases, and even macrophase separation occurs for the long-chained BCPs. The excluded volume effect of confinements helps form vesicle-like structures, which proved to be a possible method of confinement to regulate phase morphologies or segment distributions and, ultimately, the properties of materials. An analysis of the phase formation process of short-chained BCP with two hydrophilic EO segments and one hydrophobic MMA segment indicated that four stages were found in both neutral and non-neutral wall confinement, all of which present a hexagonal columnar phase. Surprisingly, when the repulsion effect of the wall to the EO segment is greater than that of the MMA segment, such BCP self assembles into a crossed columnar phase, and the intersection angle of the orientation of these two sets of cylinder arrays is 75 degrees, which can be used to produce heterogeneous nanotube arrays. For the short-chained BCP with four arms joined at MMA species and EO segments in the outer region, we found a novel method of exchanging the repulsive preference of the wall to the EO or MMA species that can control the adsorption or desorption of the lamellar phase with the interval of EO or MMA segments.

15.
Phys Chem Chem Phys ; 17(19): 12492-9, 2015 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-25898853

RESUMO

Twelve poly(ethylene oxide)-block-poly(methyl methacrylate) (PEO-b-PMMA) copolymers with lipid-like structures were designed and investigated by MesoDyn simulation. Spherical and worm-like micelles as well as bicontinuous, lamellar and defected lamellar phases were obtained. A special structure, designated B2412, with two lipid structures connected by their heads, was found to undergo four stages prior to forming a spherical micelle phase. Two possible assembly mechanisms were found via thermodynamic and dynamic process analyses; namely, the fusion and fission of micelles in dynamic equilibrium during the adjustment stage. Water can be encapsulated into these micelles, which can affect their size, particularly in low concentration aqueous solutions. The assignment of weak negative charges to the hydrophilic EO blocks resulted in a clear effect on micelle size. Surprisingly, the largest effect was observed with EO blocks with -0.5 e, wherein an ordered perfect hexagonal phase was formed. The obtained results can be applied in numerous fields of study, including adsorption, catalysis, controlled release and drug delivery.


Assuntos
Lipídeos/química , Modelos Moleculares , Polietilenoglicóis/química , Polimetil Metacrilato/química , Elétrons , Micelas , Conformação Molecular , Termodinâmica
16.
Acta Crystallogr Sect E Struct Rep Online ; 70(Pt 3): m79, 2014 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-24764945

RESUMO

In the title complex, [Cu(C6H4N5)2], the Cu(II) ion lies on an inversion center and is coordinated by two chelating 5-(pyridin-2-yl)tetra-zolide ligands in a slightly distorted square-planar coordination geometry. In the crystal, π-π stacking inter-actions, with centroid-centroid distances in the range 3.4301 (14)-3.4387 (13) Å, link the complex mol-ecules along [101].

17.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue ; 36(8): 853-859, 2024 Aug.
Artigo em Zh | MEDLINE | ID: mdl-39238410

RESUMO

OBJECTIVE: To construct Risk factor assessment table for hyperoxemia in patients after cardiopulmonary bypass heart surgery based on Delphi method, providing a basis for early prediction and assessment of the risk of hyperoxemia in patients after cardiac surgery. METHODS: A research team was established. Based on the characteristics of extracorporeal circulation cardiac surgery, the Chinese and English literature published by each database until October 2022 was retrieved and the opinions of relevant professional clinicians were combined to screen the risk factors of hyperoxemia in patients after cardiopulmonary bypass heart surgery, and the preliminary draft of the Risk factor assessment table for hyperoxemia in patients after cardiopulmonary bypass heart surgery was drawn up. The Delphi method was used to conduct two rounds of expert letter consultation to supplement and improve the initial draft and finally established the final draft of the Risk factor assessment table for hyperoxemia in patients after cardiopulmonary bypass heart surgery. RESULTS: The preliminary draft of the Risk factor assessment table for hyperoxemia in patients after cardiopulmonary bypass heart surgery was constructed according to the literature review and the opinions of relevant professional clinicians, which contained 4 dimensions and 21 items. A total of 14 experts were consulted by letter, including 5 senior titles and 9 associate senior titles. Six of them major in critical care and the other eight major in cardiovascular surgery. The effective response rates for the two rounds of questionnaire surveys were 100% and 85.71%, expert familiarity levels were 0.81 and 0.80, judgment coefficients were 0.94 and 0.92, respectively. Expert authority coefficients were both 0.86. Coefficients of variation for the importance and feasibility items in the two rounds ranged from 0.13 to 0.32 and 0.11 to 0.32, 0.06 to 0.26 and 0.06 to 0.35, respectively. The Kendall's W for importance and feasibility in the two rounds were 0.264 and 0.162, and 0.258 and 0.144 respectively, indicating statistically significant (all P < 0.05). After two rounds of expert consultations, a comprehensive evaluation and selection process resulted in the final establishment of the Risk factor assessment table for hyperoxemia in patients after cardiopulmonary bypass heart surgery, consisting of 4 dimensions and 23 items, which included general data, past history, operation-related data and postoperative data. CONCLUSIONS: The Risk factor assessment table for hyperoxemia in patients after cardiopulmonary bypass heart surgery based on the Delphi method is highly scientific and feasible, which can provide reference for clinical assessments of the risk of hyperoxemia in such patients.


Assuntos
Ponte Cardiopulmonar , Humanos , Ponte Cardiopulmonar/efeitos adversos , Fatores de Risco , Medição de Risco/métodos , Técnica Delphi , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Hiperóxia/diagnóstico , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/etiologia
18.
Front Neurol ; 15: 1451116, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39148699

RESUMO

Background: Estimated pulse wave velocity (ePWV), which measures vascular aging, is an independent predictor of cardiovascular death. Nevertheless, the relationship between ePWV and all-cause mortality among patients suffering from non-traumatic subarachnoid hemorrhages (NSAH) remains obscure. Consequently, the objective of this study is to ascertain whether ePWV exerts influence on the prognosis of individuals afflicted with NSAH. Methods: Through the Medical Information Mart for Intensive Care IV (MIMIC-IV) database, 644 eligible participants were included. The Kaplan-Meier survival curve method was employed to assess the disparity in survival status between the low and high ePWV cohorts. The Cox proportional hazard model was employed to investigate the association between ePWV and inpatient mortality among critically ill patients diagnosed with NSAH. The Restricted Cubic Spline (RCS) model was employed to examine the dose-response correlation. Subsequently, multivariate Cox regression analysis was performed to identify independent prognostic factors. Lastly, the impact of ePWV on inpatient mortality across various subgroups was evaluated through stratified analysis. Results: Participants were categorized into two groups, delineated by their ePWV levels: a low ePWV level group and a high ePWV level group. Survival analysis unveiled that individuals with high ePWV exhibited a diminished survival rate compared to their counterparts with low ePWV. Following adjustment, low ePWV was significantly linked with a reduced risk of inpatient mortality among patients with NSAH (HR = 0.54, 95% CI = 0.32-0.89, p = 0.016). Simultaneously, analysis employing the RCS model further substantiated a linear escalation in the risk of inpatient mortality with increasing ePWV values. Conclusion: Elevated ePWV levels have been identified as an independent risk factor for the rise in inpatient mortality among NSAH patients and as a significant predictor of the clinical outcome of NSAH.

19.
MAbs ; 16(1): 2394230, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39192463

RESUMO

We previously described an in vitro single-chain fragment (scFv) library platform originally designed to generate antibodies with excellent developability properties. The platform design was based on the use of clinical antibodies as scaffolds into which replicated natural complementarity-determining regions purged of sequence liabilities were inserted, and the use of phage and yeast display to carry out antibody selection. In addition to being developable, antibodies generated using our platform were extremely diverse, with most campaigns yielding sub-nanomolar binders. Here, we describe a platform advancement that incorporates Fab phage display followed by single-chain antibody-binding fragment Fab (scFab) yeast display. The scFab single-gene format provides balanced expression of light and heavy chains, with enhanced conversion to IgG, thereby combining the advantages of scFvs and Fabs. A meticulously engineered, quality-controlled Fab phage library was created using design principles similar to those used to create the scFv library. A diverse panel of binding scFabs, with high conversion efficiency to IgG, was isolated against two targets. This study highlights the compatibility of phage and yeast display with a Fab semi-synthetic library design, offering an efficient approach to generate drug-like antibodies directly, facilitating their conversion to potential therapeutic candidates.


Assuntos
Afinidade de Anticorpos , Fragmentos Fab das Imunoglobulinas , Biblioteca de Peptídeos , Anticorpos de Cadeia Única , Fragmentos Fab das Imunoglobulinas/genética , Fragmentos Fab das Imunoglobulinas/imunologia , Fragmentos Fab das Imunoglobulinas/química , Humanos , Anticorpos de Cadeia Única/genética , Anticorpos de Cadeia Única/imunologia , Anticorpos de Cadeia Única/química , Afinidade de Anticorpos/imunologia , Técnicas de Visualização da Superfície Celular/métodos , Imunoglobulina G/genética , Imunoglobulina G/imunologia , Imunoglobulina G/química
20.
Plant Biotechnol J ; 11(6): 671-80, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23521778

RESUMO

Targeted gene regulation via designed transcription factors has great potential for precise phenotypic modification and acceleration of novel crop trait development. To this end, designed transcriptional activators have been constructed by fusing transcriptional activation domains to DNA-binding proteins. In this study, a transcriptional activator from the herpes simplex virus, VP16, was used to identify plant regulatory proteins. Transcriptional activation domains were identified from each protein and fused with zinc finger DNA-binding proteins (ZFPs) to generate designed transcriptional activators. In addition, specific sequences within each transcriptional activation domain were modified to mimic the VP16 contact motif that interacts directly with RNA polymerase II core transcription factors. To evaluate these designed transcriptional activators, test systems were built in yeast and tobacco comprising reporter genes driven by promoters containing ZFP-binding sites upstream of the transcriptional start site. In yeast, transcriptional domains from the plant proteins ERF2 and PTI4 activated MEL1 reporter gene expression to levels similar to VP16 and the modified sequences displayed even greater levels of activation. Following stable transformation of the tobacco reporter system with transcriptional activators derived from ERF2, GUS reporter gene transcript accumulation was equal to or greater than those derived from VP16. Moreover, a modified ERF2 domain displayed significantly enhanced transcriptional activation compared with VP16 and with the unmodified ERF2 sequence. These results demonstrate that plant sequences capable of facilitating transcriptional activation can be found and, when fused to DNA-binding proteins, can enhance gene expression.


Assuntos
Regulação da Expressão Gênica de Plantas , Nicotiana/genética , Engenharia de Proteínas , Fatores de Transcrição/metabolismo , Ativação Transcricional/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Cromossomos de Plantas/genética , Evolução Molecular , Genes Reporter , Proteína Vmw65 do Vírus do Herpes Simples/metabolismo , Dados de Sequência Molecular , Proteínas de Plantas/química , Estrutura Terciária de Proteína , Proteoma/metabolismo , Saccharomyces cerevisiae/metabolismo , Alinhamento de Sequência , Transcrição Gênica
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