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BACKGROUND: The lateral flow immunoassay (LFIA) is widely employed as a point-of-care testing (POCT) technique. However, its limited sensitivity hinders its application in detecting biomarkers with low abundance. Recently, the utilization of nanozymes has been implemented to enhance the sensitivity of LFIA by catalyzing the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB). The catalytic performance of nanozymes plays a crucial role in influencing the sensitivity of LFIA. RESULTS: The Cornus officinalis Sieb. et Zucc-Pd@Pt (CO-Pd@Pt) nanozyme with good peroxidase-like activity was synthesized herein through a facile one-pot method employing Cornus officinalis Sieb. et Zucc extract as a reducing agent. The morphology and composition of the CO-Pd@Pt nanozyme were characterized using TEM, SEM, XRD, and XPS. As a proof of concept, the as-synthesized CO-Pd@Pt nanozyme was utilized in LFIA (CO-Pd@Pt-LFIA) for the detection of human chorionic gonadotropin (hCG). Compared to conventional gold nanoparticles-based LFIA (AuNPs-LFIA), CO-Pd@Pt-LFIA demonstrated a significant enhancement in the limit of detection (LOD, 0.08 mIU/mL), which is approximately 160 times lower than that of AuNPs-LFIA. Furthermore, experiments evaluating accuracy, precision, selectivity, interference, and stability have confirmed the practical applicability of CO-Pd@Pt-LFIA for hCG content determination. SIGNIFICANCE: The present study presents a novel approach for the synthesis of bimetallic nanozymes through environmentally friendly methods, utilizing plant extracts as both protective and reducing agents. Additionally, an easily implementable technique is proposed to enhance signal detection in lateral flow immunoassays.
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Paládio , Platina , Paládio/química , Platina/química , Imunoensaio/métodos , Humanos , Nanopartículas Metálicas/química , Limite de Detecção , Peroxidase/química , Peroxidase/metabolismo , Benzidinas/química , Catálise , OxirreduçãoRESUMO
The aryl hydrocarbon receptor (AhR) is a transcription factor that is activated by various ligands, including pollutants, microorganisms, and metabolic substances. It is expressed extensively in pulmonary and intestinal epithelial cells, where it contributes to barrier defense. The expression of AhR is pivotal in regulating the inflammatory response to microorganisms. However, dysregulated AhR expression can result in endocrine disorders, leading to immunotoxicity and potentially promoting the development of carcinoma. This review focuses on the crucial role of the AhR in facilitating and limiting the proliferation of pathogens, specifically in relation to the host cell type and the species of etiological agents involved in microbial pathogen infections. The activation of AhR is enhanced through the IDO1-AhR-IDO1 positive feedback loop, which is manipulated by viruses. AhR primarily promotes the infection of SARS-CoV-2 by inducing the expression of angiotensin-converting enzyme 2 (ACE2) and the secretion of pro-inflammatory cytokines. AhR also plays a significant role in regulating various types of T-cells, including CD4+ T cells and CD8+ T cells, in the context of pulmonary infections. The AhR pathway plays a crucial role in regulating immune responses within the respiratory and intestinal barriers when they are invaded by viruses, bacteria, parasites, and fungi. Additionally, we propose that targeting the agonist and antagonist of AhR signaling pathways could serve as a promising therapeutic approach for combating pathogen infections, especially in light of the growing prevalence of drug resistance to multiple antibiotics.
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COVID-19 , Inflamação , Receptores de Hidrocarboneto Arílico , Animais , Humanos , Fatores de Transcrição Hélice-Alça-Hélice Básicos , COVID-19/imunologia , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Inflamação/imunologia , Inflamação/metabolismo , Receptores de Hidrocarboneto Arílico/metabolismo , SARS-CoV-2/fisiologia , SARS-CoV-2/imunologia , Transdução de SinaisRESUMO
The development of nanomaterials that mimic oxidase-like activities has recently attracted an increasing amount of attention. Obtaining highly active and cost-effective oxidase mimics has posed a significant challenge in this area of research. In this study, we successfully synthesized nickel-doped ferrous disulfide nanocubes (Ni-FeS2) via a facile one-step method. Characterization by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed that Ni was predominantly distributed within the surface layer of the Ni-FeS2 nanocubes. The incorporation of nickel in density functional theory (DFT) calculations effectively reduced the d-band center of Fe, resulting in weakened adsorption to intermediates and thereby enhancing its catalytic efficiency. Moreover, we developed a novel approach based on Ni-FeS2 (the Ni-FeS2 method) for detecting reducing substances, which exhibited good sensitivity toward ascorbic acid (AA), glutathione (GSH), and cysteine (Cys). Remarkably, the established Ni-FeS2 method was successfully employed for in vitro assessment of total antioxidant capacity (TAC) in cellular and organ samples, thereby enabling discrimination between normal, senescent, and malignant cells as well as distinguishing among healthy liver tissue, cancerous liver tissue, and metastatic organs.
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Antioxidantes , Técnicas Biossensoriais , Ferro , Sulfetos , Oxirredutases , Níquel , GlutationaRESUMO
Group 2 innate lymphoid cells (ILC2s), characterized by a lack of antigen receptors, have been regarded as an important component of type 2 pulmonary immunity. Analogous to Th2 cells, ILC2s are capable of releasing type 2 cytokines and amphiregulin, thus playing an essential role in a variety of diseases, such as allergic diseases and virus-induced respiratory diseases. Interferons (IFNs), an important family of cytokines with potent antiviral effects, can be triggered by microbial products, microbial exposure, and pathogen infections. Interestingly, the past few years have witnessed encouraging progress in revealing the important role of IFNs and IFN-producing cells in modulating ILC2 responses in allergic lung inflammation and respiratory viral infections. This review underscores recent progress in understanding the role of IFNs and IFN-producing cells in shaping ILC2 responses and discusses disease phenotypes, mechanisms, and therapeutic targets in the context of allergic lung inflammation and infections with viruses, including influenza virus, rhinovirus (RV), respiratory syncytial virus (RSV), and severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2).
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COVID-19 , Interferons , Humanos , Imunidade Inata , Linfócitos , SARS-CoV-2 , Citocinas , PulmãoRESUMO
To inhibit the growth of bacteria, the DA-PPI nanozyme with enhanced peroxidase-like activity was synthesized. The DA-PPI nanozyme was obtained by depositing high-affinity element iridium (Ir) on the surface of Pd-Pt dendritic structures. The morphology and composition of DA-PPI nanozyme were characterized using SEM, TEM, and XPS. The kinetic results showed that the DA-PPI nanozyme possessed a higher peroxidase-like activity than that of Pd-Pt dendritic structures. The PL, ESR, and DFT were employed to explain the high peroxidase activity. As a proof of concept, the DA-PPI nanozyme could effectively inhibit E. coli (G-) and S. aureus (G+) due to its high peroxidase-like activity. The study provides a new idea for the design of high active nanozymes and their application in the field of antibacterial.
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The design of highly active nanozymes and the establishment of ultra-sensitive bioassays remain a challenge. Therefore, it is necessary to synthesize highly active nanozymes. In this work, a Pd-Pt-Ru (PPR) nanozyme was prepared by atomic coating of the bimetallic nanozyme Pd-Pt. The steady-state kinetics showed that the PPR nanozyme had excellent peroxidase-like activity. Based on this concept, the as-prepared PPR nanozyme was applied to the detection of ascorbic acid (AA) and hydrogen peroxide (H2O2). The linear ranges for ascorbic acid and hydrogen peroxide were 2-12 µM and 5-40 mM, respectively. The limits of detection (LOD) are 1.13 µM and 2.79 mM, respectively. Ascorbic acid was used as a typical model to assay the total antioxidant capacity (TAC) of foods and several herbs. The Fructus Corni extract showed the highest reducing ability. The corresponding extracts were applied for the green synthesis of silver nanoparticles with a size of 167 nm. This study provides a method for the design of highly active nanozymes and the expansion of their applications.
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Antioxidantes , Nanopartículas Metálicas , Peróxido de Hidrogênio , Prata , Ácido Ascórbico , PeroxidasesRESUMO
Pyroptosis is a form of cell death mediated by inflammasomes and gasdermins, and the relevance of pyroptosis to neurodegenerative diseases is currently receiving increasing attention. Alzheimer's disease (AD) is a chronic progressive neurodegenerative disease that is closely associated with neuroinflammation. Its main pathological features include ß-amyloid (Aß) deposition, Tau protein hyperphosphorylation and neuronal loss. Aß, tau-induced microglia pyroptosis and polarization leading to neuroinflammation play an important role in the pathogenesis of AD. Studying the pathogenesis and treatment of AD based on cellular pyroptosis has become a new direction in AD research. In this paper, we review the research progress of pyroptosis and will focus on the pathogenic roles of pyroptosis in AD and the role of targeted inhibition of inflammasome-dependent pyroptosis in AD treatment. These results deepen our understanding of the pathogenesis of AD and provide ideas for the development of new drugs based on the regulation of pyroptosis in AD patients.
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Ureaplasma urealyticum (U. urealyticum, Uu) is a common sexually transmitted pathogen that is responsible for diseases such as non-gonococcal urethritis, chorioamnionitis, and neonatal respiratory diseases. The rapid emergence of multidrug-resistant bacteria threatens the effective treatment of Uu infections. Considering this, vaccination could be an efficacious medical intervention to prevent Uu infection and disease. As a highly conserved molecular chaperone, DnaJ is expressed and upregulated by pathogens soon after infection. Here, we assessed the vaccine potential of recombinant Uu-DnaJ in a mouse model and dendritic cells. Results showed that intramuscular administration of DnaJ induced robust humoral- and T helper (Th) 1 cell-mediated immune responses and protected against genital tract infection, inflammation, and the pathologic sequelae after Uu infection. Importantly, the DnaJ protein also induced the maturation of mouse bone marrow-derived dendritic cells (BMDCs), ultimately promoting naïve T cell differentiation toward the Th1 phenotype. In addition, adoptive immunization of DnaJ-pulsed BMDCs elicited antigen-specific Immunoglobulin G2 (IgG2) antibodies as well as a Th1-biased cellular response in mice. These results support DnaJ as a promising vaccine candidate to control Uu infections. KEY POINTS: ⢠A novel recombinant vaccine was constructed against U. urealyticum infection. ⢠Antigen-specific humoral and cellular immune responses after DnaJ vaccination. ⢠Dendritic cells are activated by Uu-DnaJ, which results in a Th1-biased immune response.
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Infecções por Ureaplasma , Vacinas , Gravidez , Feminino , Camundongos , Animais , Ureaplasma urealyticum/genética , Infecções por Ureaplasma/prevenção & controle , Infecções por Ureaplasma/microbiologia , Células Th1 , Ativação LinfocitáriaRESUMO
Mycosis, especially superficial fungal infections (SFIs), has been a serious threat to humans in recent years. Evodiamine (EVO), as an effective component of the Traditional Chinese Medicine Evodia rutaecarpa, has good antibacterial effects and low toxicity. In order to find out the potential therapeutic agents against SFIs, a series of EVO derivatives were synthesized and systematic evaluations of antifungal activity were carried out. Among them, compound A7 exhibited great antifungal activity with the values of MIC100 were 38, 38 and 2 µg/mL, respectively, against T. rubrum, T. mentagrophytes and C. albicans, and even stronger than that of ketoconazole (KCZ) with the values of MIC100 were 106, 106 and 3 µg/mL, respectively. Further antifungal evaluations in vitro verified that compound A7 indeed had favorable antifungal activity. Moreover, compound A7 could exert excellent antifungal effect on T. rubrum-infected guinea pigs, suggesting that A7 was an attractive molecule and could be a potential lead compound for the development of anti-fungal agents, and providing a great promising therapeutic strategy for fungal disease.
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Antifúngicos , Micoses , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida albicans , Cobaias , Humanos , Testes de Sensibilidade Microbiana , Micoses/tratamento farmacológico , Quinazolinas/farmacologiaRESUMO
Natural product evodiamine is one of the most privileged scaffolds in drug discovery and is suitable for derivatization, which can be conducted quickly for structure optimization and structure-activity relationship research. In this work, a comprehensive SAR study on evodiamine scaffold with N14-3'-fluorophenyl substituted was completed, and compounds with high anti-tumor activity and good inhibitory effect on Top1 and Top2 were screened out. Tested evodiamine derivatives exhibited excellent broad-spectrum anti-tumor activity. Among them, compound 8b revealed 55.15% and 55.50% inhibition for Top1 and Top2 at 25 µM, as well as 0.16 and 0.13 µM IC50 value for MGC-803 and SGC-7901 cells, respectively; compound 9a revealed 70.50% and 71.81% inhibition for Top1 and Top2 at 25 µM, as well as 0.22 and 0.27 µM IC50 value for MGC-803 and SGC-7901 cells, respectively. The further biological evaluation showed that they could functionally induce apoptosis, significantly arrest the cell cycle at the G2/M phase, and markedly inhibit cell proliferation, migration and invasion. In addition, compound 9a performed a tumor inhibitory rate of 36.35% and showed no apparent toxicity in vivo. Overall, these optimized protocols will advance the progression of cancer chemotherapy and can be used to expand the options for screening therapeutic cancer drugs.
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Antineoplásicos , Neoplasias , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose , Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , DNA Topoisomerases Tipo II , Ensaios de Seleção de Medicamentos Antitumorais , Estrutura Molecular , Quinazolinas/química , Quinazolinas/farmacologia , Relação Estrutura-AtividadeRESUMO
Ureaplasma urealyticum and Ureaplasma parvum are the most common Ureaplasma species causing repeated or persistent infection of the urogenital tract. The host can mobilize innate and adaptive immunity to defend and eliminate pathogens. However, under certain conditions, the host's immune protection cannot completely clear Ureaplasma species. Ureaplasma species have evolved a complex and sophisticated escape mechanism in the long-term defense against host immune protection. This article summarizes the research progress on Ureaplasma species' immune escape mechanism from several aspects such as evading host autophagy mechanism, antagonizing host nutritional immunity and regulating host cell gene expression.
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Interações Hospedeiro-Parasita , Evasão da Resposta Imune , Infecções por Ureaplasma , Ureaplasma , Interações Hospedeiro-Parasita/imunologia , Humanos , Evasão da Resposta Imune/imunologia , Pesquisa/tendências , Ureaplasma/imunologia , Infecções por Ureaplasma/imunologiaRESUMO
Nucleotide exchange factor (GrpE), a highly conserved antigen, is rapidly expressed and upregulated when Ureaplasma urealyticum infects a host, which could act as a candidative vaccine if it can induce an anti-U. urealyticum immune reaction. Here, we evaluated the vaccine potential of recombinant GrpE protein adjuvanted by Freund's adjuvant (FA), to protect against U. urealyticum genital tract infection in a mouse model. After booster immunization in mice with FA, the GrpE can induced both humoral and cellular immune response after intramuscular injection into BALB/c mice. A strong humoral immune response was detected in the GrpE-immunized mice characterized by production of high titers of antigen-specific serum IgG (IgG1, IgG2a, and IgG3) antibodies. At the same time, the GrpE also induced a Th1-biased cytokine spectrum with high levels of IFN-γ and TNF-α after re-stimulation with immunogen GrpE in vitro, suggesting that GrpE could trigger the Th1 response when used for vaccination in the presence of FA. Although GrpE vaccination in the presence of a Th1-type adjuvant-induced had readily detectable Th1 responses, there wasn't increase inflammation in response to the infection. More importantly, the robust immune responses in mice after immunization with GrpE showed a significantly reduced U. urealyticum burden in cervical tissues. Histopathological analysis confirmed that tissues of GrpE-immunized BALB/c mice were protected against the pathological effects of U. urealyticum infection. In conclusion, this study preliminarily reveals GrpE protein as a promising new candidate vaccine for preventing U. urealyticum reproductive tract infection.
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Proteínas de Bactérias/imunologia , Colo do Útero/microbiologia , Proteínas de Choque Térmico/imunologia , Infecções do Sistema Genital/imunologia , Células Th1/imunologia , Infecções por Ureaplasma/imunologia , Ureaplasma urealyticum/fisiologia , Vacinas/imunologia , Animais , Células Cultivadas , Citocinas/metabolismo , Resistência à Doença , Feminino , Humanos , Imunidade Humoral , Imunização , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Group 2 innate lymphoid cells (ILC2s) are an important component of the innate immune system that execute important effector functions at barrier surfaces, such as lung and skin. Like T helper type 2 cells, ILC2s are able to release high amounts of type 2 cytokines that are essential in inducing allergic inflammation and eliminating helminth infections. The past few years have contributed to our better understanding of the interactions between ILC2s and other cells of the immune system via soluble factors or in a cell-cell contact manner. Myeloid cells, including mononuclear leukocytes and polymorphonuclear leukocytes, are excellent sensors of tissue damage and infection and can influence ILC2 responses in the process of allergic inflammation. In this review, we summarize recent insights on how myeloid cell subsets regulate ILC2 activation with focus on soluble factors in the context of allergic inflammation.
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Asma/imunologia , Dermatite Atópica/imunologia , Imunidade Inata/imunologia , Células Mieloides/imunologia , Fator de Transcrição GATA3/metabolismo , Humanos , Inflamação/imunologia , Interleucina-13/imunologia , Interleucina-4/imunologia , Interleucina-5/imunologia , Células Th2/imunologiaRESUMO
PURPOSE: To study the effect of curcumin on proliferation and invasion of the human retinoblastoma cells and its potential mechanism. METHODS: A cell line of retinoblastoma (WERI-Rb-1) was treated with various concentrations of curcumin (0-40 µM). Cell number was counted with CCK8 kit, and cell migration was assessed using the Transwell assay. Immunoblotting was performed to detect the proteins of metalloproteinase-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) as well as nuclear translocation of nuclear factor-κB (NF-κB, p65). RESULTS: Proliferation and migration of WERI-Rb-1 cells were significantly inhibited by curcumin in a concentration-dependent manner (0-40 µM). Protein expressions of MMP-2, MMP-9 and VEGF in the WERI-Rb-1 cells were also significantly inhibited by curcumin in a concentration-dependent manner (0-40 µM). Furthermore, nuclear translocation of NF-κB (p65) was significantly inhibited by curcumin in time-dependent manner (6-24 h). CONCLUSION: Curcumin inhibited proliferation and migration of WERI-Rb-1 cells, a cell line of human retinoblastoma, which might be through modulating NF-κB and its downstream proteins including VEGF, MMP-2, and MMP-9.
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Curcumina , Neoplasias da Retina , Retinoblastoma , Linhagem Celular Tumoral , Proliferação de Células , Curcumina/farmacologia , Humanos , Metaloproteinase 2 da Matriz , Metaloproteinase 9 da Matriz , NF-kappa B , Neoplasias da Retina/tratamento farmacológico , Retinoblastoma/tratamento farmacológico , Fator A de Crescimento do Endotélio VascularRESUMO
BACKGROUND: Chlamydia pneumoniae (C. pneumoniae) is pathogenic to humans, by causing pulmonary inflammation or bronchitis in both adolescents and young adults. However, the molecular signals linking C. pneumoniae components to inflammation remain elusive. This study was to investigate the effect of Chlamydia-specific Cpn0423 of C. pneumoniae on C. pneumoniae-mediated inflammation. RESULTS: Cpn0423 was detected outside of C. pneumoniae inclusions, which induced production of several cytokines including macrophage inflammatory protein-2 (MIP-2) and interleukins (ILs). Production of the Cpn0423-induced cytokines was markedly reduced in cells pretreated with NOD2-siRNA, but not with negative control oligonucleotides. Mice treated with Cpn0423 through intranasal administration exhibited pulmonary inflammation as evidenced by infiltration of inflammatory cells, increased inflammatory scores in the lung histology, recruitment of neutrophils and increased cytokines levels in the BALF. CONCLUSION: Cpn0423 could be sensed by NOD2, which was identified as an essential element in a pathway contributing to the development of C. pneumoniae -mediated inflammation.
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Proteínas de Bactérias/imunologia , Infecções por Chlamydophila/imunologia , Chlamydophila pneumoniae/imunologia , Mediadores da Inflamação/imunologia , Proteína Adaptadora de Sinalização NOD2/imunologia , Pneumonia Bacteriana/microbiologia , Animais , Proteínas de Bactérias/genética , Quimiocina CXCL2/genética , Quimiocina CXCL2/imunologia , Infecções por Chlamydophila/genética , Infecções por Chlamydophila/microbiologia , Chlamydophila pneumoniae/genética , Humanos , Interleucinas/genética , Interleucinas/imunologia , Pulmão/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína Adaptadora de Sinalização NOD2/genética , Pneumonia Bacteriana/genética , Pneumonia Bacteriana/imunologiaRESUMO
Objective To prepare the polyclonal antibody against human alpha thalassemia/mental retardation syndrome X-linked (ATRX) C-terminal and study the distribution and expression of ATRX protein in human cervical cancer tissues. Methods The antiserum was obtained from the BALB/c mice immunized with 6His-ATRX-C2193-2492 protein and then purified by the saturated ammonium sulfate precipitation and affinity chromatography. The titer of anti-ATRX polyclonal antibody was determined by ELISA. Its specificity was identified by SDS-PAGE analysis and Western blotting. The expression and location of ATRX in human cervical tissues were analyzed by immunohistochemistry. Results The titer of the polyclonal antibody against 6His-ATRX-C2193-2492 protein was about 1:12 800. The antibody could recognize 6His-ATRX-C2193-2492 protein specifically. With the polyclonal antibody, the target protein was found mainly in the nucleus of para-carcinoma tissues, and it was also expressed in the nucleus of cervical cancer tissue cells, but the expression in the latter was obviously lower. Conclusion The polyclonal antibody against 6His-ATRX-C2193-2492 protein has been produced successfully and used to detect ATRX protein in human cervical cancer tissues.
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Anticorpos/imunologia , DNA Helicases/análise , Deficiência Intelectual Ligada ao Cromossomo X/diagnóstico , Proteínas Nucleares/análise , Talassemia alfa/diagnóstico , Animais , DNA Helicases/genética , DNA Helicases/imunologia , Feminino , Humanos , Imunização , Imuno-Histoquímica , Deficiência Intelectual Ligada ao Cromossomo X/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Nucleares/genética , Proteínas Nucleares/imunologia , Proteína Nuclear Ligada ao X , Talassemia alfa/imunologiaRESUMO
Tuberculosis (TB), the leading cause of death among infectious diseases worldwide, is caused by Mycobacterium tuberculosis (M. tuberculosis). Early accurate diagnosis means earlier prevention, treatment and control of TB. To confirm efficient diagnostic antigens for M. tuberculosis, the serodiagnosis value of four recombinant proteins including Rv0220, Rv2958c, Rv2994 and Rv3347c was evaluated in this study. The specificities and sensitivities of four recombinant proteins were determined based on enzyme-linked immunosorbent assay (ELISA) by screening sera from smear-positive pulmonary TB patients (n = 92), uninfected individuals (n = 60) and patients with Mycoplasma pneumoniae (n = 32) that potentially cross-react with M. tuberculosis. The ELISAs showed that Rv0220, Rv2958c, Rv2994 and Rv3347c exhibited high specificities and sensitivities in detecting immunoglobulin G (IgG) antibody, with 98.3/91.3%, 91.7/85.9%, 93.3/89.1% and 93.3/80.4% respectively. According to the receiver-operating characteristic (ROC) analysis, the area under the ROC of the target proteins was 0.988, 0.969, 0.929 and 0.945 respectively. Western blot was established to evaluate the immunoreactivities of target proteins to mice and human sera. Results demonstrated that Rv0220, Rv2958c, Rv2994 and Rv3347c could specifically recognize TB-positive sera and the sera of mice immunized with the corresponding protein. Thus, Rv0220, Rv2958c, Rv2994 and Rv3347c were valuable potential diagnostic antigens for M. tuberculosis.
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Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Mycobacterium tuberculosis/imunologia , Testes Sorológicos/métodos , Tuberculose/diagnóstico , Animais , Antígenos de Bactérias/genética , Western Blotting , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina G/sangue , Camundongos , Mycobacterium tuberculosis/genética , Curva ROC , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Sensibilidade e EspecificidadeRESUMO
Syphilis is a multistage disease caused by the invasive spirochete Treponema pallidum subsp. pallidum, and accurate diagnosis is important for the prevention and treatment of syphilis. Here, to identify appropriate diagnostic antigens for serodiagnosis of syphilis, 6 recombinant proteins were expressed in Escherichia coli and purified, including flagellins (FlaB1 [Tp0868], FlaB2 [Tp0792], and FlaB3 [Tp0870]), Tp0463, Tp0751, and Tp1038. The sensitivities were determined by screening sera from individuals with primary (n=82), secondary (n=115), latent (n=105), and congenital (n=65) syphilis. The specificities were determined by screening sera from uninfected controls (n=30) and potentially cross-reactive infections including Lyme disease (n=30), leptospirosis (n=5), and hepatitis B (n=30). Our data showed that FlaB1, FlaB2, FlaB3, Tp0463, and Tp1038 exhibited higher overall sensitivities and specificities for detecting IgG antibody, with 95.4% and 98.9%, 92.6% and 95.8%, 95.1% and 95.8%, 92.6% and 97.9%, and 95.9% and 98.9%, respectively. In contrast, Tp0751 demonstrated only an overall sensitivity of 39.2%. For comparison, the sensitivity and specificity of Architect Syphilis TP were determined to be 98.1% and 93.7%, respectively. In addition, FlaB1, FlaB2, FlaB3, and Tp0463 demonstrated excellent performance for detecting IgM antibody in primary and congenital syphilis, with sensitivities of 76.8% and 83.1%, 72.0% and 87.7%, 74.4% and 89.2%, and 64.6% and 75.3%, respectively. These results indicate that FlaB1, FlaB2, FlaB3, and Tp0463 could be as novel diagnostic candidates for serodiagnosis of syphilis.
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Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Flagelina/imunologia , Testes Sorológicos/métodos , Sífilis/diagnóstico , Treponema pallidum/imunologia , Antígenos de Bactérias/genética , Escherichia coli/genética , Flagelina/genética , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Ureaplasma urealyticum is a major pathogen associated with many diseases. The ability of U. urealyticum to protect itself from oxidative stress is likely to be important for its pathogenesis and survival, but its oxidative stress tolerance mechanisms remain unclear. This study investigates the antioxidant activity of a ferritin-like protein from U. urealyticum. RESULTS: The uuferritin gene, which was up regulated when U. urealyticum was subjected to oxidative stress, was cloned from U. urealyticum and the corresponding recombinant protein uuferritin was purified. Uuferritin protein reduced the levels of hydroxyl radicals generated by the Fenton reaction as a consequence of its ferroxidase activity, and thus the protein protected DNA from oxidative damage. Furthermore, oxidation-sensitive Escherichia coli mutants transformed with pTrc99a-uuferritin showed significantly improved tolerance to oxidative stress compared to E. coli mutants transformed with an empty pTrc99a vector. CONCLUSIONS: The present work shows that uuferritin protein confers resistance to oxidative stress in vitro and in E. coli. The protective role of uuferritin provides a foundation for understanding the mechanisms of oxidative stress tolerance in U. urealyticum.