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1.
Protein Expr Purif ; 215: 106410, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38040273

RESUMO

Cytokeratin 19 fragment (CYFRA21-1) serves as a crucial tumor marker in the context of lung cancer patients, playing a pivotal role as a calibrator in the realm of in vitro diagnostics. Nevertheless, during practical application, it has come to light that the recombinantly synthesized full-length CYFRA21-1 antigen exhibits suboptimal stability at the requisite concentration, while the utilization of natural antigens incurs a substantial cost. To address this issue, our investigation harnessed a strategic approach whereby the soluble fragment of cytokeratin 19 (Aa244-400) was integrated into the pET32a vector, subsequently being expressed within E. coli through a fusion with the TrxA protein. This process involved induction of protein expression through 0.2 mM IPTG at 16 °C for a duration of 16 h. After induction, the target protein was purified through Ni affinity and ion exchange chromatography. Subsequent characterization of the targeted protein was executed through the SEC-HPLC technique. The attained CYFRA21-1 antigen, as generated within this study, was effectively incorporated into a chemiluminescence-based in vitro diagnostic detection kit. The results indicate that the fusion protein exhibited commendable reactivity and stability, manifesting a deviation of less than 10 % following incubation at 37 °C for 7 days. Importantly, the production yield achieved a notable magnitude of 300 mg/L, thus rendering it a cost-effective and scalable alternative to natural antigens for clinical diagnostic applications.


Assuntos
Queratina-19 , Neoplasias Pulmonares , Humanos , Queratina-19/genética , Queratina-19/análise , Escherichia coli/genética , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/análise , Proteínas
2.
Inorg Chem ; 63(25): 11802-11811, 2024 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-38861686

RESUMO

Removing trace acetylene from the ethylene stream through selective hydrogenation is a crucial process in the production of polymer-grade ethylene. However, achieving high selectivity while maintaining high activity remains a significant challenge, especially for nonprecious metal catalysts. Herein, the trade-off between activity and selectivity is solved by synergizing enhanced dispersion and hydrogen spillover. Specifically, a bubbling method is proposed for preparing SiO2-supported copper and/or bismuth carbonate with high dispersion, which is then employed to synthesize highly dispersed Bi-modified CuxC-Cu catalyst. The catalyst displays outstanding catalytic performance for acetylene selective hydrogenation, achieving acetylene conversion of 100% and ethylene selectivity of 91.1% at 100 °C. The high activity originates from the enhanced dispersion, and the exceptional selectivity is due to the enhanced spillover capacity of active hydrogen from CuxC to Cu, which is promoted by the Bi addition. The results offer an avenue to design efficient catalysts for selective hydrogenation from nonprecious metals.

3.
Inorg Chem ; 63(12): 5611-5622, 2024 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-38477101

RESUMO

The significant threat posed by the high toxicity of heavy metals and antibiotics in water pollutants has prompted a growing emphasis on the development of highly efficient removal methods for these pollutants. In this paper, flexible electrospinning polyacrylonitrile (PAN) nanofiber-supported CdBi2S4 was synthesized via a hydrothermal method, followed by amination treatment with diethylenetriamine (DETA). The as-prepared CdBi2S4/NH2-PAN nanofiber, enriched with sulfur vacancies, demonstrated outstanding visible-light trapping ability and a suitable band gap, leading to efficient separation and transport of photogenerated carriers, ultimately resulting in exceptional photocatalytic capability. The optimal 3-CdBi2S4/NH2-PAN nanofiber achieved impressive reduction rates of 92.26% for Cr(VI) and 96.45% for tetracycline hydrochloride (TCH) within 120 min, which were much higher than those for CdS/NH2-PAN, Bi2S3/NH2-PAN, and CdBi2S4/PAN nanofibers. After five cycles, the removal rate of the CdBi2S4/NH2-PAN nanofiber consistently remained above 90%. Their ease of separation and recovery from the application environment contributes to their practicality. Additionally, compared with conventional suspended particle catalyzers, the composite nanofiber exhibited remarkable flexibility and self-supporting properties.

4.
Inorg Chem ; 63(8): 3974-3985, 2024 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-38346714

RESUMO

Designing transition-metal oxides for catalytically removing the highly toxic benzene holds significance in addressing indoor/outdoor environmental pollution issues. Herein, we successfully synthesized ultrathin LayCoOx nanosheets (thickness of ∼1.8 nm) with high porosity, using a straightforward coprecipitation method. Comprehensive characterization techniques were employed to analyze the synthesized LayCoOx catalysts, revealing their low crystallinity, high surface area, and abundant porosity. Catalytic benzene oxidation tests demonstrated that the La0.029CoOx-300 nanosheet exhibited the most optimal performance. This catalyst enabled complete benzene degradation at a relatively low temperature of 220 °C, even under a high space velocity (SV) of 20,000 h-1, and displayed remarkable durability throughout various catalytic assessments, including SV variations, exposure to water vapor, recycling, and long time-on-stream tests. Characterization analyses confirmed the enhanced interactions between Co and doped La, the presence of abundant adsorbed oxygen, and the extensive exposure of Co3+ species in La0.029CoOx-300 nanosheets. Theoretical calculations further revealed that La doping was beneficial for the formation of oxygen vacancies and the adsorption of more hydroxyl groups. These features strongly promoted the adsorption and activation of oxygen, thereby accelerating the benzene oxidation processes. This work underscores the advantages of doping rare-earth elements into transition-metal oxides as a cost-effective yet efficient strategy for purifying industrial exhausts.

5.
Small ; 19(25): e2300456, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36932874

RESUMO

In the context of thin-film nanocomposite membranes with interlayer (TFNi), nanoparticles are deposited uniformly onto the support prior to the formation of the polyamide (PA) layer. The successful implementation of this approach relies on the ability of nanoparticles to meet strict requirements regarding their sizes, dispersibility, and compatibility. Nevertheless, the synthesis of covalent organic frameworks (COFs) that are well-dispersed, uniformly morphological, and exhibit improved affinity to the PA network, while preventing agglomeration, remains a significant challenge. In this work, a simple and efficient method is presented for the synthesis of well-dispersed, uniformly morphological, and amine-functionalized 2D imine-linked COFs regardless of the ligand composition, group type, or framework pore size, by utilizing a polyethyleneimine (PEI) shielded covalent self-assembly strategy. Subsequently, the as-prepared COFs are incorporated into TFNi for the recycling of pharmaceutical synthetic organic solvents. After optimization, the membrane exhibits a high rejection rate and a favorable solvent flux, making it a reliable method for efficient organic recovery and the concentration of active pharmaceutical ingredient (API) from the mother liquor through an organic solvent forward osmosis (OSFO) process. Notably, this study represents the first investigation of the impact of COF nanoparticles in TFNi on OSFO performance.

6.
Inorg Chem ; 62(33): 13544-13553, 2023 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-37561968

RESUMO

In catalytic oxidation reactions, the presence of environmental water poses challenges to the performance of Pt catalysts. This study aims to overcome this challenge by introducing hydroxyl groups onto the surface of Pt catalysts using the pyrolysis reduction method. Two silica supports were employed to investigate the impact of hydroxyl groups: SiO2-OH with hydroxyl groups and SiO2-C without hydroxyl groups. Structural characterization confirmed the presence of Pt-Ox, Pt-OHx, and Pt0 species in the Pt/SiO2-OH catalysts, while only Pt-Ox and Pt0 species were observed in the Pt/SiO2-C catalysts. Catalytic performance tests demonstrated the remarkable capacity of the 0.5 wt % Pt/SiO2-OH catalyst, achieving complete conversion of benzene at 160 °C under a high space velocity of 60,000 h-1. Notably, the catalytic oxidation capacity of the Pt/SiO2-OH catalyst remained largely unaffected even in the presence of 10 vol % water vapor. Moreover, the catalyst exhibited exceptional recyclability and stability, maintaining its performance over 16 repeated cycles and a continuous operation time of 70 h. Theoretical calculations revealed that the construction of Pt-OHx sites on the catalyst surface was beneficial for modulating the d-band structure, which in turn enhanced the adsorption and activation of reactants. This finding highlights the efficacy of decorating the Pt surface with hydroxyl groups as an effective strategy for improving the water resistance, catalytic activity, and long-term stability of Pt catalysts.

7.
Sensors (Basel) ; 23(10)2023 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-37430629

RESUMO

The quick and accurate characterization of commercial electrochemical double-layer capacitor (EDLC) cells, especially their capacitance and direct-current equivalent series internal resistance (DCESR), is of great significance for the design, maintenance, and monitoring of EDLCs used in areas of energy, sensors, electric power, construction machinery, rail transit, automobile transportation, and military. In this study, the capacitance and DCESR of three commercial EDLC cells with similar performance were determined and compared by following the three commonly-used standards of IEC 62391, Maxwell, and QC/T741-2014, which are significantly different in test procedures and calculation methods. The analysis of the test procedures and results demonstrated that the IEC 62391 standard has the disadvantages of a large testing current, long testing time, and a complex and inaccurate DCESR calculation, whereas the Maxwell standard has the disadvantages of a large testing current, a small capacitance, and large DCESR testing results, and furthermore the QC/T 741 standard has the disadvantages of a high resolution requirement for the equipment and small DCESR results. Therefore, an improved method was proposed to determine the capacitance and DCESR of EDLC cells by short-time constant voltage charging and discharging interruption methods, respectively, with the advantages of high accuracy, low equipment requirements, short testing time, and the easy calculation of DCESR over the original three standards.

8.
Nanotechnology ; 32(14): 145718, 2021 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-33333494

RESUMO

Defect engineering has been widely used in experiments to modulate the electrocatalytic properties of molybdenum disulfide (MoS2). However, the effect of vacancy concentration on the vacancy distribution, electronic properties, and hydrogen evolution reaction (HER) activity remains elusive. Herein, we perform density functional theory (DFT) studies to investigate defective MoS2 with different numbers of sulfur vacancies. In the case of low S-vacancy concentration, the vacancies prefer to agglomerate rather than being dispersed, while at the higher-vacancy concentration, the combination of local point defect and clustered vacancy chain is preferred. The coupling between S-vacancies leads to decreased band gap and increased Mo-H adsorption strength with increasing vacancy concentration. The optimal HER activity is identified to occur below vacancy concentration of 12.50%. Our work provides an atomic-level understanding about the role of S-vacancies in the HER performance of MoS2, and offers useful guidelines for the design of defective MoS2 and other TMDs electrocatalysts.

9.
Anal Chem ; 91(22): 14530-14537, 2019 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-31617350

RESUMO

In this study, an accurately and digitally regulated allosteric nanoswitch based on the conformational control of two DNA hairpins was developed. By switching between UV irradiation and blue light conditions, the second molecular beacon (H#2) would bind/separate with a repression sequence (RES) via the introduced PTG molecules (a photosensitive azobenzene derivative), resulting in the target aptamer sequence in the first molecular beacon (H#1) not being able/being able to hold the stem-loop configuration, hence losing/regaining the ability to bind with the target. Importantly, we successfully monitor conformation changes of the nanoswitch by an elegant mathematical model for connecting Ki (the dissociation constant between RES and H#2) with Kd (the overall equilibrium constant of the nanoswitch binding the target), hence realizing "observing" DNA structure across dimensions from "structural visualization" to digitization and, accurately, digitally regulating DNA structure from digitization to "structural visualization".


Assuntos
Aptâmeros de Nucleotídeos/química , DNA/química , Nanoestruturas/química , Compostos Azo/química , Compostos Azo/efeitos da radiação , DNA/metabolismo , DNA/efeitos da radiação , Sequências Repetidas Invertidas , Ligantes , Modelos Químicos , Conformação de Ácido Nucleico/efeitos da radiação , Trombina/metabolismo , Raios Ultravioleta
10.
J Virol ; 91(19)2017 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-28701399

RESUMO

Herpes simplex virus (HSV) infection is restricted to epithelial cells and neurons and is controlled by CD8 T cells. These cells both traffic to epithelial sites of recurrent lytic infection and to ganglia and persist at the dermal-epidermal junction for up to 12 weeks after lesion resolution. We previously showed that cutaneous lymphocyte-associated antigen (CLA), a functional E-selectin ligand (ESL), is selectively expressed on circulating HSV-2-specific CD8 T cells. CLA/ESL mediates adhesion of T cells to inflamed vascular endothelium. Later stages in T-cell homing involve chemokines (Ch) and lymphocyte chemokine receptors (ChR) for vascular wall arrest and diapedesis. Several candidate ChR have been implicated in skin homing. We measured cell surface ChR on HSV-specific human peripheral blood CD8 T cells and extended our studies to HSV-1. We observed preferential cell surface expression of CCR10 and CXCR3 by HSV-specific CD8 T cells compared to CD8 T cells specific for control viruses, Epstein-Barr virus (EBV) and cytomegalovirus (CMV), and compared to bulk memory CD8 T cells. CXCR3 ligand mRNA levels were selectively increased in skin biopsy specimens from persons with recurrent HSV-2, while the mRNA levels of the CCR10 ligand CCL27 were equivalent in lesion and control skin. Our data are consistent with a model in which CCL27 drives baseline recruitment of HSV-specific CD8 T cells expressing CCR10, while interferon-responsive CXCR3 ligands recruit additional cells in response to virus-driven inflammation.IMPORTANCE HSV-2 causes very localized recurrent infections in the skin and genital mucosa. Virus-specific CD8 T cells home to the site of recurrent infection and participate in viral clearance. The exit of T cells from the blood involves the use of chemokine receptors on the T-cell surface and chemokines that are present in infected tissue. In this study, circulating HSV-2-specific CD8 T cells were identified using specific fluorescent tetramer reagents, and their expression of several candidate skin-homing-associated chemokine receptors was measured using flow cytometry. We found that two chemokine receptors, CXCR3 and CCR10, are upregulated on HSV-specific CD8 T cells in blood. The chemokines corresponding to these receptors are also expressed in infected tissues. Vaccine strategies to prime CD8 T cells to home to HSV lesions should elicit these chemokine receptors if possible to increase the homing of vaccine-primed cells to sites of infection.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Quimiocina CCL27/imunologia , Herpes Simples/imunologia , Herpesvirus Humano 2/imunologia , Ativação Linfocitária/imunologia , Receptores CCR10/imunologia , Receptores CXCR3/imunologia , Antígenos de Diferenciação de Linfócitos T/imunologia , Linfócitos T CD8-Positivos/metabolismo , Quimiocina CCL27/genética , Citomegalovirus/imunologia , Feminino , Citometria de Fluxo , Herpes Simples/virologia , Herpesvirus Humano 4/imunologia , Humanos , Memória Imunológica/imunologia , Masculino , Glicoproteínas de Membrana/imunologia , RNA Mensageiro/genética , Receptores CCR10/biossíntese , Receptores CCR10/genética , Receptores CXCR3/biossíntese , Receptores CXCR3/genética , Pele/virologia
11.
J Immunol ; 196(5): 2205-2218, 2016 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-26810224

RESUMO

The Alphaherpesvirinae subfamily includes HSV types 1 and 2 and the sequence-divergent pathogen varicella zoster virus (VZV). T cells, controlled by TCR and HLA molecules that tolerate limited epitope amino acid variation, might cross-react between these microbes. We show that memory PBMC expansion with either HSV or VZV enriches for CD4 T cell lines that recognize the other agent at the whole-virus, protein, and peptide levels, consistent with bidirectional cross-reactivity. HSV-specific CD4 T cells recovered from HSV-seronegative persons can be explained, in part, by such VZV cross-reactivity. HSV-1-reactive CD8 T cells also cross-react with VZV-infected cells, full-length VZV proteins, and VZV peptides, as well as kill VZV-infected dermal fibroblasts. Mono- and cross-reactive CD8 T cells use distinct TCRB CDR3 sequences. Cross-reactivity to VZV is reconstituted by cloning and expressing TCRA/TCRB receptors from T cells that are initially isolated using HSV reagents. Overall, we define 13 novel CD4 and CD8 HSV-VZV cross-reactive epitopes and strongly imply additional cross-reactive peptide sets. Viral proteins can harbor both CD4 and CD8 HSV/VZV cross-reactive epitopes. Quantitative estimates of HSV/VZV cross-reactivity for both CD4 and CD8 T cells vary from 10 to 50%. Based on these findings, we hypothesize that host herpesvirus immune history may influence the pathogenesis and clinical outcome of subsequent infections or vaccinations for related pathogens and that cross-reactive epitopes and TCRs may be useful for multi-alphaherpesvirus vaccine design and adoptive cellular therapy.


Assuntos
Alphaherpesvirinae/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Reações Cruzadas/imunologia , Infecções por Herpesviridae/imunologia , Apresentação de Antígeno/imunologia , Antígenos Virais/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Citocinas/metabolismo , Epitopos de Linfócito T/imunologia , Infecções por Herpesviridae/genética , Infecções por Herpesviridae/virologia , Herpesvirus Humano 1/imunologia , Herpesvirus Humano 2/imunologia , Humanos , Peptídeos/imunologia , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Proteínas Virais/imunologia
12.
J Ind Microbiol Biotechnol ; 45(4): 253-269, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29399712

RESUMO

In this paper, a novel riboregulator Switch System of Gene Expression including an OFF-TO-ON switch and an ON-TO-OFF switch was designed to regulate the expression state of target genes between "ON" and "OFF" by switching the identifiability of ribosome recognition site (RBS) based on the thermodynamic stability of different RNA-RNA hybridizations between RBS and small noncoding RNAs. The proposed riboregulator switch system was employed for the fermentative production of succinic acid using an engineered strain of E. coli JW1021, during which the expression of mgtC gene was controlled at "ON" state and that of pepc and ecaA genes were controlled at the "OFF" state in the lag phase and switched to the "OFF" and "ON" state once the strain enters the logarithmic phase. The results showed that using the strain of JW1021, the yield and productivity of succinic acid can reach 0.91 g g-1 and 3.25 g L-1 h-1, respectively, much higher than those using the strains without harboring the riboregulator switch system.


Assuntos
Escherichia coli/metabolismo , Fermentação , Regulação Bacteriana da Expressão Gênica , Microbiologia Industrial/métodos , Ácido Succínico/metabolismo , Reatores Biológicos , Farmacorresistência Bacteriana , Escherichia coli/genética , Expressão Gênica , Glucose/química , Canamicina/química , Cinética , Hibridização de Ácido Nucleico , Oligonucleotídeos/genética , RNA/química , Termodinâmica
13.
Metab Eng ; 42: 85-97, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28603040

RESUMO

Artificial control of bio-functions through regulating gene expression is one of the most important and attractive technologies to build novel living systems that are useful in the areas of chemical synthesis, nanotechnology, pharmacology, cell biology. Here, we present a novel real-time control system of gene regulation that includes an enhancement element by introducing duplex DNA aptamers upstream promoter and a repression element by introducing a RNA aptamer upstream ribosome binding site. With the presence of ligands corresponding to the DNA aptamers, the expression of the target gene can be potentially enhanced at the transcriptional level by strengthening the recognition capability of RNAP to the recognition region and speeding up the separation efficiency of the unwinding region due to the induced DNA bubble around the thrombin-bound aptamers; while with the presence of RNA aptamer ligand, the gene expression can be repressed at the translational level by weakening the recognition capability of ribosome to RBS due to the shielding of RBS by the formed aptamer-ligand complex upstream RBS. The effectiveness and potential utility of the developed gene regulation system were demonstrated by regulating the expression of ecaA gene in the cell-free systems. The realistic metabolic engineering application of the system has also tested by regulating the expression of mgtC gene and thrombin cDNA in Escherichia coli JD1021 for controlling metabolic flux and improving thrombin production, verifying that the real-time control system of gene regulation is able to realize the dynamic regulation of gene expression with potential applications in bacterial physiology studies and metabolic engineering.


Assuntos
Aptâmeros de Nucleotídeos/genética , Aptâmeros de Nucleotídeos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Engenharia Metabólica/métodos , Humanos
14.
Genes Immun ; 17(7): 412-418, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27853144

RESUMO

Variation at HLA and KIR loci is associated with the severity of viral infections. To assess associations of genital HSV-2 infection with human HLA and KIR genetic loci, we measured the frequencies of genital herpes simplex virus (HSV) DNA detection and of genital lesions in HSV-2 seropositive persons. We followed 267 HSV-2 seropositive persons who collected daily genital swabs and recorded lesions for ⩾30 days. All persons were laboratory-documented as HIV-seronegative, and all were Caucasian by self-report. HSV detection rate and lesion frequency were compared by genotype using Poisson regression. Overall, HSV was detected on 19.1% of days and lesions on 11.6% of days. The presence of HLA-A*01 was directly associated with HSV detection frequency, whereas the presence of HLA-C*12 was inversely associated with HSV detection frequency. The presence of HLA-A*01 was directly associated with lesion rate, while HLA-A*26, -C*01 and -DQB1*0106 were associated with decreased lesions. We observed an interaction between the absence of both 2DS4del and HLA-Bw4 and higher lesion rate. Heterozygosity of HLA was also associated with reduced lesion frequency. Immune control of genital HSV infection relies on multiple interacting immunogenetic elements, including epistatic interactions between HLA and KIR.


Assuntos
Genes MHC da Classe II , Genes MHC Classe I , Herpes Genital/virologia , Herpesvirus Humano 2/fisiologia , Receptores KIR/metabolismo , Eliminação de Partículas Virais , Adulto , Idoso , Estudos de Coortes , Feminino , Genótipo , Herpes Genital/patologia , Heterozigoto , Humanos , Desequilíbrio de Ligação , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Recidiva , Adulto Jovem
15.
J Ind Microbiol Biotechnol ; 43(4): 505-16, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26711444

RESUMO

In this study, a novel engineering Escherichia coli strain (CBMG111) with the expression of mgtCB gene was constructed for the enhanced fermentative production of succinic acid by utilizing the synergetic effect of mgtC gene to improve the growth of strains at the environment of low Mg(2+) concentration and mgtB to enhance the transport of Mg(2+) into cells. After the effect of the expression of the individual genes (mgtA, mgtB, mgtC) on the growth of E. coli was clarified, the fermentative production of succinic acid by CBMG111 was studied with the low-price mixture of Mg(OH)2 and NH3·H2O as the alkaline neutralizer and the biomass hydrolysates as the carbon sources, which demonstrated that the expression of mgtCB gene can significantly increase the productivity of succinic acid (2.97 g L(-1) h(-1)) compared with that by using the engineering strain with the overexpression of mgtA gene.


Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Reatores Biológicos , Escherichia coli/genética , Mutação , Ácido Succínico/metabolismo , Antiporters/genética , Antiporters/metabolismo , Escherichia coli/metabolismo , Fermentação , Magnésio/metabolismo
16.
J Infect Dis ; 212(7): 1022-31, 2015 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-25784732

RESUMO

BACKGROUND: The live, attenuated varicella vaccine strain (vOka) is the only licensed therapeutic vaccine. Boost of varicella zoster virus (VZV)-specific cellular immunity is a likely mechanism of action. We examined memory CD4(+) T-cell responses to each VZV protein at baseline and after zoster vaccination. METHODS: Serial blood samples were collected from 12 subjects vaccinated with Zostavax and immunogenicity confirmed by ex vivo VZV-specific T-cell and antibody assays. CD4(+) T-cell lines enriched for VZV specificity were generated and probed for proliferative responses to every VZV protein and selected peptide sets. RESULTS: Zoster vaccination increased the median magnitude (2.3-fold) and breadth (4.2-fold) of VZV-specific CD4(+) T cells one month post-vaccination. Both measures declined by 6 months. The most prevalent responses at baseline included VZV open reading frames (ORFs) 68, 4, 37, and 63. After vaccination, responses to ORFs 40, 67, 9, 59, 12, 62, and 18 were also prevalent. The immunogenicity of ORF9 and ORF18 were confirmed using peptides, defining a large number of discrete CD4 T-cell epitopes. CONCLUSIONS: The breadth and magnitude of the VZV-specific CD4(+) T-cell response increase after zoster vaccination. In addition to glycoprotein E (ORF68), we identified antigenic ORFs that may be useful components of subunit vaccines.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Vacina contra Herpes Zoster/imunologia , Herpes Zoster/imunologia , Herpesvirus Humano 3/imunologia , Vacinação , Sequência de Aminoácidos , Epitopos de Linfócito T/imunologia , Herpes Zoster/prevenção & controle , Humanos , Imunidade Celular , Pessoa de Meia-Idade , Dados de Sequência Molecular , Vacinas Atenuadas/imunologia
17.
Appl Microbiol Biotechnol ; 99(20): 8575-86, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26092756

RESUMO

For making a complex synthetic gene network function as designed, the parameters in the network have to be extensively tuned. In this study, a simple and general approach to rapidly tune gene networks in Escherichia coli was developed, which uses the hypermutable simple sequence repeats embedded in the spacer region between the ribosome binding site and the initiation codon. It was found that the change of sequence length and compositions of the repeated base pairs in 5'UTR contributes together to the changeable expression levels of the target gene. The mechanism of this phenomenon is that the transcriptional process makes greater impact on the expression level when compared to the translational process, which is utilized to successfully predict sample gene expression levels over a 50-fold range. The utility of the approach to regulate heterologous ecaA and pepc gene expression in the engineered E. coli for improving succinic acid yield and production has also been demonstrated. When the expression level of ecaA gene was 3.53-fold of the control and the expression level of pepc gene was 1.06-fold of the control, the highest yield of succinic acid and productivity were achieved, which was 0.87 g g(-1) and 2.01 g L(-1) h(-1), respectively.


Assuntos
Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Engenharia Metabólica/métodos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Ácido Succínico/metabolismo , Regiões 5' não Traduzidas
18.
medRxiv ; 2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38352384

RESUMO

The skin at the site of HSV-2 reactivation is enriched for HSV-2-specific T cells. To evaluate whether an immunotherapeutic vaccine could elicit skin-based memory T cells, we studied skin biopsies and HSV-2-reactive CD4+ T cells from peripheral blood mononuclear cells (PBMCs) by T cell receptor ß (TRB) sequencing before and after vaccination with a replication-incompetent whole virus HSV-2 vaccine candidate (HSV529). The representation of HSV-2-reactive CD4+ TRB sequences from PBMCs in the skin TRB repertoire increased after the first vaccine dose. We found sustained expansion after vaccination of unique, skin-based T-cell clonotypes that were not detected in HSV-2-reactive CD4+ T cells isolated from PBMCs. In one participant a switch in immunodominance occurred with the emergence of a T cell receptor (TCR) αß pair after vaccination that was not detected in blood. This TCRαß was shown to be HSV-2-reactive by expression of a synthetic TCR in a Jurkat-based NR4A1 reporter system. The skin in areas of HSV-2 reactivation possesses an oligoclonal TRB repertoire that is distinct from the circulation. Defining the influence of therapeutic vaccination on the HSV-2-specific TRB repertoire requires tissue-based evaluation.

19.
Org Lett ; 25(31): 5774-5778, 2023 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-37503938

RESUMO

In this work, an N-substituted diketopyrrolopyrrole (DPP) derivative Ph-DPP was synthesized, showing interaction toward Lewis alkaline anions such as F-. The typical electron-transfer-dominated anion-π interaction product Ph-DPP•- and unexpected isomer product i-Ph-DPP were both observed, and their formation mechanism was studied by density functional theory calculations, suggesting that a deprotonation initiation route is favored, which gives interesting insight for understanding the debatable role of F- in such non-covalent intermolecular interactions.

20.
Clin Pharmacol Drug Dev ; 12(11): 1069-1075, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37246720

RESUMO

Celecoxib is a sulfanilamide nonsteroidal anti-inflammatory drug that can selectively inhibit cyclooxygenase-2 to inhibit prostaglandin production, achieving anti-inflammatory and analgesic effects. This study investigated the pharmacokinetics, safety, and bioequivalence of a single oral dose of celecoxib capsule (the test or reference preparation) in healthy volunteers under fasting and fed conditions. A single-center, randomized, open, single-dose, double-cycle crossover self-control design was conducted: 40 healthy volunteers were enrolled in the fasting and fed groups, respectively. A completely randomized method was used, with one group taking the test celecoxib preparation (T) and the other taking the reference celecoxib preparation (R). During the administration period, the safety of the drug was evaluated simultaneously, and venous blood was collected at the corresponding time points. The concentration of celecoxib in plasma was measured by liquid chromatography-tandem mass spectrometry. The main pharmacokinetic parameters were logarithmically converted and analyzed for variance. The 90% confidence interval for the bioavailability of the T compared to the R was calculated using maximum drug plasma concentration, area under the plasma concentration-time curve from time zero to the last quantifiable concentration point, and area under the plasma concentration-time curve from time zero to infinity for a single oral dose in volunteers, and the data obtained were all between 80% and 125%, indicating that the T and R have bioequivalence and good safety during fasting and fed administration.


Assuntos
Anti-Inflamatórios , Celecoxib , População do Leste Asiático , Humanos , Anti-Inflamatórios/farmacocinética , Celecoxib/farmacocinética , Voluntários Saudáveis , Equivalência Terapêutica
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