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1.
Hortic Res ; 11(3): uhae016, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38495032

RESUMEN

Artificially enhancing photosynthesis is critical for improving crop yields and fruit qualities. Nanomaterials have demonstrated great potential to enhance photosynthetic efficiency; however, the mechanisms underlying their effects are poorly understood. This study revealed that the electron transfer pathway participated in nitrogen-doped carbon dots (N-CDs)-induced photosynthetic efficiency enhancement (24.29%), resulting in the improvements of apple fruit qualities (soluble sugar content: 11.43%) in the orchard. We also found that N-CDs alleviated mterf5 mutant-modulated photosystem II (PSII) defects, but not psa3 mutant-modulated photosystem I (PSI) defects, suggesting that the N-CDs-targeting sites were located between PSII and PSI. Measurements of chlorophyll fluorescence parameters suggested that plastoquinone (PQ), the mobile electron carrier in the photosynthesis electron transfer chain (PETC), was the photosynthesis component that N-CDs targeted. In vitro experiments demonstrated that plastoquinone-9 (PQ-9) could accept electrons from light-excited N-CDs to produce the reduced plastoquinone 9 (PQH2-9). These findings suggested that N-CDs, as electron donors, offer a PQ-9-involved complement of PETC to improve photosynthesis and thereby fruit quality. Our study uncovered a mechanism by which nanomaterials enhanced plant photosynthesis and provided some insights that will be useful in the design of efficient nanomaterials for agricultural/horticultural applications.

2.
Int J Biol Macromol ; 264(Pt 2): 130689, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38458287

RESUMEN

African Swine Fever Virus (ASFV) is a highly contagious pathogen posing a serious threat to the global swine industry. Despite this, there is currently no effective vaccine against this virus. Within ASFV's core shell structure, p37, a product of polyprotein pp220, shares sequence similarity with SUMO-1 proteases. Localization studies show p37 in various nuclear regions during early infection, shifting to the cytoplasm later on. Research indicates active export of p37 from the nucleus, mediated by CRM1-dependent and -independent pathways. Hydrophobic amino acids in p37 are crucial for these pathways, highlighting their importance throughout the ASFV replication cycle. Additionally, p37 serves as the first nucleocytoplasmic shuttle protein encoded by ASFV, participating in the intranuclear material transport process during ASFV infection of host cells. In this study, we successfully screened five murine monoclonal antibodies targeting p37. Through the truncated expression method, we identified four dominant antigenic epitopes of p37 for the first time. Furthermore, utilizing alanine scanning technology, we determined the key amino acid residues for each epitope. This research not only provides essential information for a deeper understanding of the protein's function but also establishes a significant theoretical foundation for the design and development of ASFV vaccines.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Porcinos , Animales , Ratones , Anticuerpos Monoclonales , Proteínas Virales/química , Fiebre Porcina Africana/prevención & control
3.
Biomed Pharmacother ; 171: 116099, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38171244

RESUMEN

Chronic myelogenous leukemia (CML) is characterized by the constitutive activation of BCR-ABL tyrosine kinase. Imatinib was approved for CML therapy, however, BCR-ABL-dependent drug resistance, especially BCR-ABL-T315I mutation, restricts its clinical application. In this study, we reported anthraquinone lactone AS1041, a synthesized derivative of marine natural compound Aspergiolide A, showed anti-leukemia effect in vitro and in vivo by promoting cell senescence. Mechanistic study revealed the pro-senescence effect of AS1041 was dependent on oxidative stress-induced DNA damage, and the resultant activation of P53/P21 and P16INK4a/Rb. Also, AS1041 promoted ubiquitin proteasome system (UPS)-mediated BCR-ABL degradation, which also contributed to AS1041-induced senescence. In vivo, AS1041-induced senescence promoted tumor growth inhibition. In summary, the in vitro and in vivo antitumor effect of AS1041 suggests it can serve as a pro-senescence agent for alternative antileukemia therapy and imatinib-resistant cancer therapy by enhancing cellular oxidative stress and BCR-ABL degradation.


Asunto(s)
Antraquinonas , Resistencia a Antineoplásicos , Leucemia Mielógena Crónica BCR-ABL Positiva , Humanos , Mesilato de Imatinib/farmacología , Apoptosis , Proliferación Celular , Proteínas de Fusión bcr-abl/metabolismo , Estrés Oxidativo , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Daño del ADN , Inhibidores de Proteínas Quinasas/farmacología
4.
J Biol Chem ; 299(8): 104990, 2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-37392850

RESUMEN

Cycloartenyl ferulate (CF) is abundant in brown rice with multiple biologic functions. It has been reported to possess antitumor activity; however, the related mechanism of action of CF has not been clarified. Herein, we unexpectedly uncover the immunological regulation effects of CF and its molecular mechanism. We discovered that CF directly enhanced the killing capacity of natural killer (NK) cells for various cancer cells in vitro. In vivo, CF also improved cancer surveillance in mouse models of lymphoma clearance and metastatic melanoma dependent on NK cells. In addition, CF promoted anticancer efficacy of the anti-PD1 antibody with improvement of tumor immune microenvironment. Mechanistically, we first unveiled that CF acted on the canonical JAK1/2-STAT1 signaling pathway to enhance the immunity of the NK cells by selectively binding to interferon γ receptor 1. Collectively, our results indicate that CF is a promising immunoregulation agent worthy of attention in clinical application in the future. Due to broad biological significance of interferon γ, our findings also provide a capability to understand the diverse functions of CF.


Asunto(s)
Ácidos Cumáricos , Células Asesinas Naturales , Neoplasias , Receptores de Interferón , Animales , Ratones , Interferón gamma/inmunología , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/inmunología , Neoplasias/inmunología , Microambiente Tumoral , Ácidos Cumáricos/farmacología , Receptores de Interferón/inmunología , Receptor de Interferón gamma
5.
Int J Mol Sci ; 24(2)2023 Jan 09.
Artículo en Inglés | MEDLINE | ID: mdl-36674804

RESUMEN

The nuclear receptors-liver X receptors (LXR α and ß) are potential therapeutic targets in cardiovascular and neurodegenerative diseases because of their key role in the regulation of lipid homeostasis and inflammatory processes. Specific oxy(phyto)sterols differentially modulate the transcriptional activity of LXRs providing opportunities to develop compounds with improved therapeutic characteristics. We isolated oxyphytosterols from Sargassum fusiforme and synthesized sidechain oxidized sterol derivatives. Five 24-oxidized sterols demonstrated a high potency for LXRα/ß activation in luciferase reporter assays and induction of LXR-target genes APOE, ABCA1 and ABCG1 involved in cellular cholesterol turnover in cultured cells: methyl 3ß-hydroxychol-5-en-24-oate (S1), methyl (3ß)-3-aldehydeoxychol-5-en-24-oate (S2), 24-ketocholesterol (S6), (3ß,22E)-3-hydroxycholesta-5,22-dien-24-one (N10) and fucosterol-24,28 epoxide (N12). These compounds induced SREBF1 but not SREBP1c-mediated lipogenic genes such as SCD1, ACACA and FASN in HepG2 cells or astrocytoma cells. Moreover, S2 and S6 enhanced cholesterol efflux from HepG2 cells. All five oxysterols induced production of the endogenous LXR agonists 24(S)-hydroxycholesterol by upregulating the CYP46A1, encoding the enzyme converting cholesterol into 24(S)-hydroxycholesterol; S1 and S6 may also act via the upregulation of desmosterol production. Thus, we identified five novel LXR-activating 24-oxidized sterols with a potential for therapeutic applications in neurodegenerative and cardiovascular diseases.


Asunto(s)
Enfermedades Neurodegenerativas , Fitosteroles , Humanos , Receptores X del Hígado , Esteroles/farmacología , Receptores Nucleares Huérfanos/genética , Hidroxicolesteroles , Enfermedades Neurodegenerativas/tratamiento farmacológico , Colesterol
6.
Anal Biochem ; 660: 114953, 2023 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-36243135

RESUMEN

Human papillomaviruse type 16 (HPV16) is a high-risk serotype. As the main protective antigen protein, L1 protein is also the target protein for diagnosis. A simple label free electrochemical immunosensor (ECIS) was fabricated for ultrasensitive detection of HPV16 L1 protein in this work. Quasi-spherical Ag@Au core-shell nanoparticles on graphene oxide (Ag@AuNPs-GO) was developed as current response amplifier and characterized by UV-Vis Spectroscopy, Transmission Electron Microscopy and energy dispersive X-ray spectroscopy. Staphylococcal protein A was decorated on the modified electrode and utilized to immobilized the Fc portion of the monoclonal antibody specific for HPV16 L1 protein. Cyclic Voltammetry, Differential Pulse Voltammetry and Electrochemical Impedance Spectroscopy were used to verify the electrochemical performance and interfacial kinetic property. The increased concentration of HPV16 L1 protein led to slow electron transport and linearly decreased differential pulse voltammetry peak current with a detection limit of 0.002 ng mL-1 and a wide linear relationship in the range of 0.005-400 ng mL-1at a regression coefficient (R2) of 0.9948. Furthermore, this ECIS demonstrated acceptable accuracy with good reproducibility, stability and selectivity, suggesting a promising immunological strategy for HPV typing and early screening.


Asunto(s)
Alphapapillomavirus , Técnicas Biosensibles , Grafito , Nanopartículas del Metal , Humanos , Oro/química , Técnicas Biosensibles/métodos , Nanopartículas del Metal/química , Inmunoensayo/métodos , Reproducibilidad de los Resultados , Grafito/química , Técnicas Electroquímicas/métodos , Límite de Detección
7.
J Sci Food Agric ; 103(6): 2997-3005, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36448538

RESUMEN

BACKGROUND: Astaxanthin is a carotenoid with strong antioxidant property. In addition, it has anti-cancer, anti-tumor, anti-inflammatory and many other functions. The micro-organisms that mainly produce astaxanthin are Haematococcus pluvialis and Phaffia rhodozyma. Compared with H. pluvialis, P. rhodozyma has shorter fermentation cycle and easier to control culture conditions, but the yield of astaxanthin in P. rhodozyma is low. This article studied how to improve the astaxanthin production of P. rhodozyma. RESULTS: The results showed that when 10 mL L-1 soybean oil was added to the culture medium, astaxanthin production increased significantly, reaching 7.35 mg L-1 , which was 1.4 times that of the control group, and lycopene and ß-carotene contents also increased significantly. Through targeted metabolite analysis, the fatty acids in P. rhodozyma significantly increased under the soybean oil stimulation, especially the fatty acids closely related to the formation of astaxanthin esters, included palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1n9), linoleic acid (C18:2n6), α-linolenic acid (C18:3n3) and γ-linolenic acid (C18:3n6), thereby increasing the astaxanthin esters content. CONCLUSION: It showed that the addition of soybean oil can promote the accumulation of astaxanthin by promoting the increase of astaxanthin ester content. © 2022 Society of Chemical Industry.


Asunto(s)
Basidiomycota , Aceite de Soja , Aceite de Soja/metabolismo , Xantófilas/metabolismo , Basidiomycota/metabolismo , Ácidos Grasos/metabolismo
8.
Luminescence ; 37(8): 1300-1308, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35637545

RESUMEN

Bacitracin zinc (BAC), a polypeptide antibiotic, is utilized as a feed additive due to its ability to promote growth in animals. However, the abuse of BAC can lead to a great threat to food safety. Therefore, there is an urgent need to develop a rapid and sensitive detection method. In this study, a monoclonal antibody (mAb) against BAC with excellent sensitivity and specificity was obtained. For the first time, quantum dots (QDs) were conjugated with the prepared mAb against BAC and rabbit anti-mouse antibody to fabricate a direct and an indirect competitive fluorescence-linked immunosorbent assay (dc-FLISA and ic-FLISA) to detect BAC. The IC50 of dc-FLISA and ic-FLISA were 0.28 ng/ml and 0.17 ng/ml, respectively. The limits of detection were 0.0016 ng/ml and 0.001 ng/ml, respectively, and the detection ranges were 0.0016-46.50 ng/ml and 0.001-35.65 ng/ml, respectively. In addition, the recovery rate of the two methods ranged from 93.5% to 112.0%, and the coefficient of variation (CV) was less than 10%. Therefore, the methods developed in this work have the merits of low cost, simple operation, and high sensitivity, which provide an effective analytical tool for BAC residue detection in feed samples.


Asunto(s)
Puntos Cuánticos , Animales , Anticuerpos Monoclonales/química , Bacitracina , Ensayo de Inmunoadsorción Enzimática/métodos , Inmunoadsorbentes/química , Límite de Detección , Puntos Cuánticos/química , Conejos
9.
Antioxidants (Basel) ; 11(4)2022 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-35453471

RESUMEN

Natural bromophenols are important secondary metabolites in marine algae. Derivatives of these bromophenol are potential candidates for the drug development due to their biological activities, such as antioxidant, anticancer, anti-diabetic and anti-inflammatory activity. In our present study, we have designed and synthesized a series of new methylated and acetylated bromophenol derivatives from easily available materials using simple operation procedures and evaluated their antioxidant and anticancer activities on the cellular level. The results showed that 2.,3-dibromo-1-(((2-bromo-4,5-dimethoxybenzyl)oxy)methyl)-4,5-dimethoxybenzene (3b-9) and (oxybis(methylene))bis(4-bromo-6-methoxy-3,1-phenylene) diacetate (4b-3) compounds ameliorated H2O2-induced oxidative damage and ROS generation in HaCaT keratinocytes. Compounds 2.,3-dibromo-1-(((2-bromo-4,5-dimethoxybenzyl)oxy)methyl)-4,5-dimethoxybenzene (3b-9) and (oxybis(methylene) )bis(4-bromo-6-methoxy-3,1-phenylene) diacetate (4b-3) also increased the TrxR1 and HO-1 expression while not affecting Nrf2 expression in HaCaT. In addition, compounds (oxybis(methylene)bis(2-bromo-6-methoxy-4,1-phenylene) diacetate (4b-4) inhibited the viability and induced apoptosis of leukemia K562 cells while not affecting the cell cycle distribution. The present work indicated that some of these bromophenol derivatives possess significant antioxidant and anticancer potential, which merits further investigation.

10.
Front Cell Infect Microbiol ; 12: 816238, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35252031

RESUMEN

Hepatitis C virus (HCV) infection is a global public health threat. Reaching the World Health Organization's objective for eliminating viral hepatitis by 2030 will require a precise disease diagnosis. While immunoassays and qPCR play a significant role in detecting HCV, rapid and accurate point-of-care testing is important for pathogen identification. This study establishes a reverse transcription recombinase-aided amplification-lateral flow dipstick (RT-RAA-LFD) assay to detect HCV. The intact workflow was completed within 30 min, and the detection limit for synthesized C/E1 plasmid gene-containing plasmid was 10 copies/µl. In addition, the test showed good specificity, with no cross-reactivity observed for hepatitis A virus, hepatitis B virus, HIV, syphilis, and human papillomavirus virus. Using extracted RNAs from 46 anti-HCV antibody-positive samples, RT-RAA-LFD showed 100% positive and negative concordance rates with qPCR. In summary, the RT-RAA-LFD assay established in this study is suitable for the rapid clinical detection of HCV at the community level and in remote areas.


Asunto(s)
Hepatitis C , Transcripción Reversa , Hepacivirus/genética , Hepatitis C/diagnóstico , Humanos , Técnicas de Amplificación de Ácido Nucleico , Recombinasas/metabolismo , Sensibilidad y Especificidad
11.
Mar Drugs ; 19(11)2021 Nov 16.
Artículo en Inglés | MEDLINE | ID: mdl-34822512

RESUMEN

Angiogenesis, including the growth of new capillary blood vessels from existing ones and the malignant tumors cells formed vasculogenic mimicry, is quite important for the tumor metastasis. Anti-angiogenesis is one of the significant therapies in tumor treatment, while the clinical angiogenesis inhibitors usually exhibit endothelial cells dysfunction and drug resistance. Bis(2,3,6-tribromo-4,5-dihydroxybenzyl)ether (BTDE), a marine algae-derived bromophenol compound, has shown various biological activities, however, its anti-angiogenesis function remains unknown. The present study illustrated that BTDE had anti-angiogenesis effect in vitro through inhibiting human umbilical vein endothelial cells migration, invasion, tube formation, and the activity of matrix metalloproteinases 9 (MMP9), and in vivo BTDE also blocked intersegmental vessel formation in zebrafish embryos. Moreover, BTDE inhibited the migration, invasion, and vasculogenic mimicry formation of lung cancer cell A549. All these results indicated that BTDE could be used as a potential candidate in anti-angiogenesis for the treatment of cancer.


Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Microalgas , Fenoles/farmacología , Células A549/efectos de los fármacos , Inhibidores de la Angiogénesis/química , Animales , Organismos Acuáticos , Proliferación Celular/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Humanos , Neovascularización Fisiológica/efectos de los fármacos , Fenoles/química
12.
Antioxidants (Basel) ; 10(9)2021 Sep 09.
Artículo en Inglés | MEDLINE | ID: mdl-34573068

RESUMEN

Excessive reactive oxygen species (ROS) promotes the oxidative stress of keratinocytes, eventually causing cell damage. The natural bromophenol bis (2,3,6-tribromo-4,5-dihydroxybenzyl) ether (BTDE) from marine red algae has been reported to have a varied bioactivity; however, its antioxidant effect has yet to be investigated systemically. Our present work aimed to explore the antioxidant effect of BTDE both on the molecular and cellular models and also to illustrate the antioxidant mechanisms. Our results showed that BTDE could effectively scavenge ABTS free radicals and protect HaCaT cells from damage induced by H2O2. Mechanism studies in HaCaT cells demonstrated that BTDE attenuated hydrogen peroxide (H2O2)-induced ROS production, reduced the malondialdehyde (MDA) level, decreased the oxidized glutathione (GSSG)/glutathione (GSH) ratio, and increased the antioxidant enzyme superoxide dismutase (SOD). Moreover, BTDE could inhibit the expression of Kelch-like epichlorohydrin-associated protein 1 (Keap1) and increase the expression of both nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream proteins TrXR1, HO-1, and NQO1. BTDE also activated the upstream signaling pathway of Nrf2 such as AKT pathway, while not activating the ERK or AMPKα pathways. In general, BTDE is a promising antioxidant to protect HaCaT cells against oxidative damage via Nrf2-mediated pathways.

13.
Int J Biol Macromol ; 180: 203-211, 2021 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-33737177

RESUMEN

African swine fever virus (ASFV), a re-emerging DNA virus, causes a highly contagious disease for domestic pigs. It is running rife worldwide and threatening the global swine industry. Protein p54 is an attractive candidate for ASFV diagnostic and vaccine design. In this work, we designed a peptide to mimic the N-terminal domain (NTD) of ASFV p54 and pretested it with sera from ASFV-infected pigs. The peptide could be well recognized by the sera, implying that the NTD of p54 contained some potential linear B cell epitopes. Then, the conjugates of the peptide with bovine serum albumin were used as the immunogen to generate monoclonal antibodies (mAbs). A total of six mAbs specific to the NTD of ASFV p54 protein were developed. Five of them well reacted with ASFV HLJ/18 strain and recognized a same linear B cell epitope 5FFQPV9. Furthermore, epitope 5FFQPV9 could be well recognized by ASFV-positive sera from natural infected pigs, suggesting that it was a natural linear B-cell epitope. Conservation analysis indicated that epitope 5FFQPV9 were highly conserved among ASFV epidemic isolates belonging to genotype I and II. Alanine-scanning mutagenesis further revealed that the residues (6F to 9V) of epitope 5FFQPV9 were the core binding sites for antibody recognition. This is the first research to characterize specific mAbs against NTD of p54 protein. These findings may help further understand the function of p54 protein and the improvement of ASFV diagnosis.


Asunto(s)
Virus de la Fiebre Porcina Africana/inmunología , Anticuerpos Monoclonales/inmunología , Anticuerpos Antivirales/inmunología , Antígenos Virales/inmunología , Dominios Proteicos/inmunología , Proteínas Estructurales Virales/inmunología , Virus de la Fiebre Porcina Africana/genética , Secuencia de Aminoácidos , Animales , Sitios de Unión/inmunología , Epítopos de Linfocito B/inmunología , Genotipo , Células HEK293 , Humanos , Imitación Molecular , Péptidos/química , Filogenia , Albúmina Sérica Bovina , Porcinos , Transfección , Proteínas Estructurales Virales/química , Proteínas Estructurales Virales/genética
14.
Int J Biol Macromol ; 174: 587-595, 2021 Mar 31.
Artículo en Inglés | MEDLINE | ID: mdl-33493568

RESUMEN

Persistent infection with high-risk mucosal human papillomavirus (HPV) types has much association with the development of cervical cancer. The major capsid protein L1 has been confirmed to be a major candidate antigen for the development of vaccines. Here, the HPV18 L1 protein was successfully expressed and purified, then nine anti-HPV18 L1 monoclonal antibodies were prepared. Four neutralizing monoclonal antibodies (NmAbs) were identified by using hemagglutination inhibition assay and pseudovirus based neutralization assay. The results of Dot-ELISA, Western blot and indirect immunofluorescence assay showed that the neutralizing antibodies could cross-react with HPV16/18/45/31/33/58/35/39 L1. The mimotopes on HPV18/45 L1 proteins were identified and analyzed by using both phage display and Bioinformatics tool. The B cell epitopes 43-54 aa and 116-126 aa of HPV18 L1 protein, the B cell epitope 381-389 aa of HPV45 L1 protein, and the mimotopes epitope of HPV45 L1 protein were identified by peptide-ELISA and competitive ELISA. The results of PyMOL and Pepitope server analysis indicated that epitopes recognized by NmAbs 7F4, 5A6, 3G11, and 2F5 are located on the surface of L1 VLPs. The results of this study enriched the library of HPV neutralizing antibodies, revealed the mechanism of antibody neutralization, might open new perspectives on the antibody-antigen reaction and have important implications for the development of novel HPV vaccines.


Asunto(s)
Alphapapillomavirus/inmunología , Anticuerpos Neutralizantes/análisis , Proteínas de la Cápside/administración & dosificación , Epítopos/inmunología , Animales , Anticuerpos Monoclonales/análisis , Proteínas de la Cápside/química , Proteínas de la Cápside/inmunología , Línea Celular , Epítopos de Linfocito B/inmunología , Femenino , Células HEK293 , Papillomavirus Humano 18/inmunología , Humanos , Inmunización , Imitación Molecular , Pruebas de Neutralización , Biblioteca de Péptidos
15.
J Org Chem ; 84(7): 4451-4457, 2019 04 05.
Artículo en Inglés | MEDLINE | ID: mdl-30865441

RESUMEN

The core structure of marine natural products aspergiolides A (1a) and B (1b) was achieved via a concise, two-step procedure with satisfactory yield. Based on this protocol, a natural products mimic library containing 25 structural simplified analogues of 1a was then constructed. Several prepared analogues showed potential cytotoxic activity against five different tumor cell lines, and compound 7bb, in particular, exhibited cytotoxicity comparable to that of 1a.


Asunto(s)
Antraquinonas/química , Células A549 , Antraquinonas/síntesis química , Antraquinonas/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Antineoplásicos/farmacología , Línea Celular Tumoral , Ensayos de Selección de Medicamentos Antitumorales , Células HL-60 , Células HeLa , Humanos , Células K562 , Modelos Moleculares
16.
Int J Biol Macromol ; 130: 300-306, 2019 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-30811967

RESUMEN

Porcine reproductive and respiratory syndrome virus (PRRSV) nucleocapsid protein (N) is the immunodominant region of PRRSV viral proteins. However, B cell epitopes present on N protein have not been well characterized. In this study, The ORF7 gene was amplified by RT-PCR and inserted into the expression vector pET-28a, the constructed pET-28a-N was transformed into Escherichia coli BL21. After expression, purification and identification, the recombined N protein was used as the target to generate monoclonal antibody (mAb). Strains of hybridoma cells secreting anti-N mAbs were obtained by the hybridoma technique. Three of them (named 1G4, 1C6, 3D11) were specifically reacted with PRRSV by IPMA and IFA. To identify the B-cell epitopes within PRRSV N protein, six serial overlapping synthesized peptides (P1-P6) covering the whole region of N were used to define the epitopes recognized by 1G4, 1C6, 3D11. Importantly, after the identification of dot-ELISA and indirect ELISA, we found that 1-15aa was a new epitope which had never been reported before and that it was highly conserved among some HP-PRRSV isolates of type 2 PRRSV. The results of this study might open new perspectives on the detection of PRRSV and have important implications for studing the structure of the N protein.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Mapeo Epitopo , Epítopos de Linfocito B/inmunología , Virus del Síndrome Respiratorio y Reproductivo Porcino/inmunología , Proteínas Virales/inmunología , Secuencia de Aminoácidos , Anticuerpos Monoclonales/química , Línea Celular Tumoral , Humanos , Modelos Moleculares , Conformación Proteica
17.
Int J Biol Macromol ; 118(Pt A): 1354-1361, 2018 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-29959021

RESUMEN

Persistent infection with human papillomavirus type16 (HPV16) has much association with the development of cervical cancer. L1 is the major capsid protein of HPV, it has been well investigated as a potential vaccine candidate. However, B cell epitopes present on L1 have not been well characterized. To identify the potential B-cell antigenic epitopes within HPV16 L1 protein, sixteen serial overlapping truncations (H1-H16) covering the whole region were expressed in E. coli and used in mice immunization. The mice antisera were tested in ELISA binding, IFA and HI assays. Finally, four fragments (H2, H4, H11, H12) were found to contain B cell epitopes of HPV16 L1 protein in ELISA and IFA assays, three fragments (H2, H3, H9) might contain neutralizing epitopes of HPV16 L1 protein in HI assay. Among them, H11 and H12 fragments contain B cell epitopes have never been reported before, and H3 was found as hemagglutination inhibition epitope for the first time. This work provides new insights to B cell epitopes on HPV16 L1 protein. Several new epitopes were identified and may provide some guidance for HPV16 subunit vaccine design. The results of this study might open new perspectives on the antibody-antigen reaction and have important implications for the development of epitopes-based protective HPV16 vaccines.


Asunto(s)
Proteínas de la Cápside/inmunología , Mapeo Epitopo/métodos , Epítopos de Linfocito B/inmunología , Papillomavirus Humano 16/inmunología , Proteínas Oncogénicas Virales/inmunología , Animales , Femenino , Humanos , Ratones Endogámicos BALB C
18.
Mar Drugs ; 15(11)2017 Nov 04.
Artículo en Inglés | MEDLINE | ID: mdl-29113054

RESUMEN

AS1041 is a novel synthesized anthraquinone lactone derivative of marine natural compound aspergiolide A (ASP-A) with new structure skeleton and marked cytotoxicity in cancer cells. To study its cytotoxicity in detail, we evaluated its activity on human K562 chronic myelogenous leukemia cells and investigated the related molecule mechanisms. AS1041 significantly inhibited the proliferation and colony formation of K562 cells. Moreover, AS1041 arrested cell cycle progression at G2/M phase in a concentration-dependent manner, and also caused concentration- and time-dependent induction of apoptosis. In addition, the molecular mechanisms investigation showed that AS1041 did not localize in the cellular nucleus and did not affect topoisomerases I or II. However, AS1041 could inactivate extracellular signal-regulated kinase (ERK) and contribute to AS1041-induced apoptosis. We concluded that AS1041 was cytotoxic to K562 leukemia cells and the cytotoxicity related to the cell cycle arrest, apoptosis induction, and ERK inhibition. These results implied that AS1041 was a novel derivative of ASP-A with significant cytotoxicity to chronic myelogenous leukemia cells and may have therapeutic potential for the treatment of cancer and leukemia.


Asunto(s)
Antraquinonas/química , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Lactonas/química , Antraquinonas/farmacología , Antineoplásicos/farmacología , Proliferación Celular , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Humanos , Células K562 , Lactonas/farmacología , Estructura Molecular
19.
J Nat Prod ; 79(7): 1783-90, 2016 07 22.
Artículo en Inglés | MEDLINE | ID: mdl-27359163

RESUMEN

Six new pyridone alkaloids, named penipyridones A-F (1-6), were isolated from the fermentation broth of an Antarctic moss-derived fungus, Penicillium funiculosum GWT2-24. Their structures were elucidated from extensive NMR and MS data. Although they possess the same major chromophore and some of them presented almost mirror ECD spectra, their absolute configurations were found to be uniformly S, as evidenced by X-ray single-crystal diffraction analysis, stereocontrolled total synthesis, and chemical conversions. TDDFT-ECD calculations of compounds 3 and 6 revealed that subtle conformational changes are responsible for the significantly different ECD curves. None of the compounds were cytotoxic (IC50 > 50 µM), while compounds 1, 2, 5, and 7 elicited lipid-lowering activity in HepG2 hepatocytes.


Asunto(s)
Alcaloides/aislamiento & purificación , Hipolipemiantes/aislamiento & purificación , Penicillium/química , Piridonas/aislamiento & purificación , Alcaloides/química , Alcaloides/farmacología , Regiones Antárticas , Cristalografía por Rayos X , Células Hep G2 , Humanos , Hipolipemiantes/química , Hipolipemiantes/farmacología , Conformación Molecular , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Piridonas/química , Piridonas/farmacología
20.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 32(4): 442-5, 2016 Apr.
Artículo en Chino | MEDLINE | ID: mdl-27053606

RESUMEN

OBJECTIVE: To predict the B cell epitopes of human papillomavirus type 16 (HPV-16) L1 protein. METHODS: After fetching the sequence of HPV-16 L1 from the protein data bank of NCBI, we used Protean software of DNAStar package to analyze the secondary structures, flexibility, hydrophilicity, surface accessibility and antigenicity of the protein. Then average antigen index (AI) of dominant regions was calculated using Wu Yuzhang's method. The potential B cell epitopes of HPV-16 L1 were predicted based on a comprehensive consideration of the above parameters. Finally, the homologies of the epitopes were analyzed with BLAST online. RESULTS: The B cell epitopes of HPV-16 L1 might exist at amino acids NO. 51-58, 87-97, 214-220, 290-296, 335-341, 351-366, 408-418, 430-442 and 475-496. Analysis of homologies indicated that the dominant B cell epitopes of the HPV-16 L1 protein might present at amino acids NO. 51-58, 335-341, 351-366, 408-418, 430-442 and 475-496. The sequences, such as 51(PIKKPNNN)58, 351(SETTYKNTNFKEYLRH)366, 408(PPPGGTLEDTY)418 and 430(KHTPPAPKEDPLK)442, were unique to HPV-16 L1, while amino acids 475(KAKPKFTLGKRKATPTTSSTST)496 were identical to amino acids in HPV-16 E1, and the amino acids 335(DTTRSTN)341 were identical to amino acids in other types of HPV L1. CONCLUSION: The B cell epitopes of HPV-16 L1 were predicted using multiple schemes. The results will provide a foundation for the further study and development of broadly protective HPV-16 vaccines.


Asunto(s)
Proteínas de la Cápside/química , Proteínas de la Cápside/inmunología , Epítopos de Linfocito B/inmunología , Papillomavirus Humano 16/inmunología , Proteínas Oncogénicas Virales/química , Proteínas Oncogénicas Virales/inmunología , Infecciones por Papillomavirus/virología , Proteínas de la Cápside/genética , Mapeo Epitopo , Epítopos de Linfocito B/genética , Papillomavirus Humano 16/química , Papillomavirus Humano 16/genética , Humanos , Proteínas Oncogénicas Virales/genética , Infecciones por Papillomavirus/genética , Infecciones por Papillomavirus/inmunología
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