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Parkinson's disease (PD) is a severe neurodegenerative disease that disturbs human health. In the laboratory researches about PD, the mice model induced by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was widely used. However, there has been controversy about the model effectiveness to simulate PD symptoms and pathology, and the time-varying development of behavioral and pathological characteristic after MPTP treatment remains unclear. In order to solve these problems, we designed a series of experiments to evaluate this PD model at different time points. We constructed the subacute PD mouse model by intraperitoneal injection of MPTP for 5 consecutive days. The rotarod test, open field test and the immunohistochemical staining of tyrosine hydroxylase were conducted at -5, 1, 5, 7, 14, 21 and 28 days after the last injection of MPTP. The results showed that 5 days after the last MPTP administration, typical motor disorders with significant balance function damage in rotarod test began to appear and remained stable throughout the entire experiment. Simultaneously, we also observed the loss of tyrosine hydroxylase (TH) positive cells in the substantia nigra compacta and reduction of TH content in the striatum but this pathological change in the substantia nigra compacta reversed 21 days after injection. Besides, the spontaneous movement of mice in open field test remained unchanged by MPTP. This research indicated the time-dependence of MPTP neurotoxicity that impair the motor function and histological features and confirmed the symptom occurrence time after MPTP injection, which provides a reference for the future research about MPTP-induced PD.
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Precise quantification of human cells in preclinical animal models by a sensitive and specific approach is warranted. The probe-based quantitative PCR (qPCR) assay as a sensitive and swift approach is suitable for the quantification of human cells by targeting human-specific DNA sequences. In this study, we developed an efficient qPCR assay targeting human-specific DNA in ST6GALNAC3 (termed ST6GAL-qPCR) for the quantification of human cells in preclinical animal models. ST6GAL-qPCR probe was synthesized with FAM and non-fluorescent quencher-minor groove binder conjugated to the 5' and 3' end of the probe, respectively. Genomic DNA from human, rhesus monkeys, cynomolgus monkeys, New Zealand White rabbits, SD rats, C57BL/6, and BALB/c mice were utilized for analyzing the specificity and sensitivity of the ST6GAL-qPCR assay. The ST6GAL-qPCR assay targeted human-specific DNA was cloned to pUCM-T vector and released by EcoR I/Hind III digestion for generating a calibration curve. Cell mixing experiment was performed to validate the ST6GAL-qPCR assay by analysis of 0.1%, 0.01%, and 0.001% of human leukocytes mixed with murine thymocytes. The ST6GAL-qPCR assay detected human DNA rather than DNA from the tested animal species. The amplification efficiency of the ST6GAL-qPCR assay was 93% and the linearity of calibration curve was R2 = 0.999. The ST6GAL-qPCR assay detected as low as 5 copies of human-specific DNA and is efficient to specially amplify as low as 30-pg human DNA in the presence of 1 µg of DNA from the tested species, respectively. The ST6GAL-qPCR assay was able to quantify as low as 0.01% of human leukocytes within murine thymocytes. This ST6GAL-qPCR assay can be used as an efficient approach for the quantification of human cells in preclinical animal models.
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PURPOSE: To explore the essential roles of phosphorylation in mediating the proliferation of T. gondii in its cell lytic life. METHODS: We profiled the phosphoproteome data of T. gondii residing in HFF cells for 2 h and 6 h, representing the early- and late-stages of proliferation (ESP and LSP) within its first generation of division. RESULTS: We identified 70 phosphoproteins, among which 8 phosphoproteins were quantified with the phosphorylation level significantly regulated. While only two of the eight phosphoproteins, GRA7 and TGGT1_242070, were significantly down-regulated at the transcriptional level in the group of LSP vs. ESP. Moreover, GO terms correlated with host membrane component were significantly enriched in the category of cellular component, suggesting phosphoprotein played important roles in acquiring essential substance from host cell via manipulating host membrane. Further GO analysis in the categories of molecular function and biological process and pathway analysis revealed that the cellular processes of glucose and lipid metabolism were regulated by T. gondii phosphoproteins such as PMCAA1, LIPIN, Pyk1 and ALD. Additionally, several phosphoproteins were enriched at the central nodes in the protein-protein interaction network, which may have essential roles in T. gondii proliferation including GAP45, MLC1, fructose-1,6-bisphosphate aldolase, GRAs and so on. CONCLUSION: This study revealed the main cellular processes and key phosphoproteins crucial for the intracellular proliferation of T. gondii, which would provide clues to explore the roles of phosphorylation in regulating the development of tachyzoites and new insight into the mechanism of T. gondii development in vitro.
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Fenómenos Biológicos , Toxoplasma , Animales , Toxoplasma/fisiología , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Fosforilación , Proliferación CelularRESUMEN
The mechanisms responsible for stem growth in peanut (Arachis hypogaea L.) cultivars with varying plant heights remain unclear, despite the significant impact of plant height on peanut yield. Therefore, this study aimed to investigate the underlying mechanisms of peanut stem growth using phenotypic, physiological, transcriptomic, and metabolomic analyses. The findings revealed that the tallest cultivar, HY33, exhibited the highest rate of stem growth and accumulated the most stem dry matter, followed by the intermediate cultivar, SH108, while the dwarf cultivar, Df216, displayed the lowest values. Furthermore, SH108 exhibited a higher harvest index, as well as superior pod and kernel yields compared to both HY33 and Df216. Transcriptome and metabolome analyses identified differentially expressed genes (DEGs) and differentially expressed metabolites (DEMs) associated with phenylpropanoid and flavonoid biosynthesis. Notably, downregulated DEGs in Df216/HY33 and Df216/SH108 included phenylalanine ammonia-lyase (PAL), caffeoyl-CoA O-methyltransferase (COMT), and ferulate-5-hydroxylase (F5H), while downregulated DEMs included p-coumaryl alcohol, chlorogenic acid, and L-epicatechin. Compared to HY33, the reduced activities of PAL, COMT, and F5H resulted in a decreased stem lignin content in Df216. Additionally, downregulated DEGs involved in gibberellin (GA) and brassinosteroid (BR) biosynthesis were identified in Df216/HY33, which contributed to the lowest levels of GA1, GA3, and BR contents in Df216. The results suggest that the dwarf phenotype arises from impaired GA and BR biosynthesis and signaling, resulting in a slower stem growth rate and reduced lignin accumulation.
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Arachis , Transcriptoma , Transcriptoma/genética , Arachis/metabolismo , Lignina/metabolismo , Perfilación de la Expresión Génica , Metabolómica , Regulación de la Expresión Génica de las PlantasRESUMEN
T and B lymphocytes are crucial players in cellular and humoral immune responses. The development, activation and differentiation of T and B lymphocytes are regulated by the best characterized PI3K-PI (3,4,5) P3-AKT phosphoinositide signalling pathway. As a branch of the phosphoinositide signalling pathway, the lipid phosphatase INPP4B inhibits AKT activation through degrading the phosphoinositide signalling messenger PI (3,4) P2. However, the role of Inpp4b in T and B lymphocytes remains elusive. Here, we reported that Inpp4b was highly expressed in human and murine T- and B-1 lymphocytes. Despite its higher expression in T lymphocytes, neither T cell development and homeostasis nor in vitro T cell activation and CD4+ T cell differentiation were altered upon loss of Inpp4b. Interestingly, combined direct phenotype analysis of Inpp4b conventional knockout mice and adoptive transfer studies revealed that ablation of Inpp4b intrinsically reduced peritoneal B-1 cells rather B-2 cells. Moreover, Inpp4b deficiency led to impaired thymus independent (TI) and thymus dependent (TD) antigens-induced antibody production. Further in vitro analysis revealed that CD40-mediated B cell proliferation was impaired upon ablation of Inpp4b. Our findings reveal that Inpp4b is required in regulating B-1 cell numbers and B cell-mediated antibody production.
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Subgrupos de Linfocitos B , Proteínas Proto-Oncogénicas c-akt , Humanos , Ratones , Animales , Proteínas Proto-Oncogénicas c-akt/metabolismo , Formación de Anticuerpos , Monoéster Fosfórico Hidrolasas/genética , Monoéster Fosfórico Hidrolasas/metabolismo , Antígenos , Fosfatidilinositoles , Recuento de CélulasRESUMEN
ABSTRACT Introduction In a scenario of continuous improvement of telecommunications network infrastructure and the maturation of related industrial chains, sports-oriented cell phone applications can meet the needs of different types of user groups due to their characteristics - close to the human body, able to collect users' health, favorite sports, location, and other personal characteristic data in real-time. With the support of cloud computing, big data, and other techniques, the regularity of users' sports health can be analyzed. Objective Explore the effect of obese college students' use of cell phone apps on their exercise routines. Methods 53 obese college students volunteered for the research project. They were investigated by a questionnaire about their use of apps to aid exercise and health. Then, the participants' exercise status was monitored, including weekly exercise time, exercise frequency, and exercise items, among other pertinent data. Finally, physical health data collected before and after the experiment were analyzed and statistically compared. Results Some obese college students did not acquire the habit of adhering to physical exercise, their psychological resilience and ability to fight frustration proved deficient, and the effect of an exercise assisted by cell phone apps left to be desired. However, for some obese college students who determined to exercise, the regular moderate exercise showed an improvement in physical health. Conclusion Exercise-focused health maintenance apps can stimulate obese college students' enthusiasm for exercise, making the exercise process more enjoyable, controlled, and scientific. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.
RESUMO Introdução Num cenário de melhoria contínua da infraestrutura da rede de telecomunicações e o amadurecimento das cadeias industriais relacionadas, os aplicativos de celulares voltados ao esporte podem atender às necessidades de diferentes tipos de grupos de usuários devido às suas próprias características - próximas ao corpo humano, capazes de coletar a saúde dos usuários, esportes prediletos, localização e outros dados de características pessoais em tempo real. Com o suporte da computação em nuvem, big data e outras técnicas, pode-se analisar a regularidade da saúde esportiva dos usuários. Objetivo Explorar o efeito do uso de aplicativos de celulares por estudantes universitários obesos em suas rotinas de exercícios físicos. Métodos 53 estudantes universitários obesos foram voluntários do projeto de pesquisa. Foram investigados por um questionário sobre a utilização de aplicativos voltados para auxiliar o exercício físico e a saúde. Em seguida, a situação de exercício físico dos participantes foi monitorada, incluindo tempo de exercício semanal, frequência de exercícios, itens de exercícios, entre outros dados pertinentes. Finalmente, os dados de saúde física coletados antes e depois do experimento foram analisados e comparados estatisticamente. Resultados Alguns estudantes universitários obesos não adquiriram o hábito de aderirem aos exercícios físicos, sua resistência psicológica e sua capacidade de combate à frustração mostraram-se deficientes, e o efeito do exercício assistido por aplicativos de celulares deixou a desejar. Porém, para alguns estudantes universitários obesos com determinação em se exercitar, o exercício regular e moderado evidenciou uma melhora sobre a saúde física. Conclusão Os aplicativos focados em exercícios para a manutenção da saúde podem estimular o entusiasmo dos estudantes universitários obesos no exercício físico, tornando o processo de exercício mais prazeroso, controlado e científico. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.
RESUMEN Introducción En un escenario de mejora continua de la infraestructura de las redes de telecomunicaciones y de maduración de las cadenas industriales relacionadas, las aplicaciones de telefonía móvil orientadas al deporte pueden satisfacer las necesidades de distintos tipos de grupos de usuarios gracias a sus propias características: cercanas al cuerpo humano, capaces de recoger en tiempo real los datos de salud, deportes favoritos, localización y otras características personales de los usuarios. Con el apoyo de la computación en nube, big data y otras técnicas, se puede analizar la regularidad de la salud deportiva de los usuarios. Objetivo Explorar el efecto del uso de aplicaciones para teléfonos móviles por estudiantes universitarios obesos en sus rutinas de ejercicio. Métodos 53 estudiantes universitarios obesos participaron como voluntarios en el proyecto de investigación. Fueron investigados mediante un cuestionario sobre el uso de apps destinadas a ayudar al ejercicio físico y la salud. A continuación, se realizó un seguimiento del estado de ejercicio de los participantes, incluido el tiempo de ejercicio semanal, la frecuencia de ejercicio, los elementos de ejercicio, entre otros datos pertinentes. Por último, se analizaron y compararon estadísticamente los datos de salud física recogidos antes y después del experimento. Resultados Algunos universitarios obesos no adquirieron el hábito de realizar ejercicio físico, su resistencia psicológica y su capacidad para luchar contra la frustración resultaron deficientes, y el efecto del ejercicio asistido por aplicaciones de telefonía móvil dejó que desear. Sin embargo, para algunos estudiantes universitarios obesos con determinación para hacer ejercicio, el ejercicio regular y moderado evidenció una mejora en la salud física. Conclusión Las aplicaciones para el mantenimiento de la salud centradas en el ejercicio pueden estimular el entusiasmo de los estudiantes universitarios obesos por el ejercicio físico, haciendo que el proceso de ejercicio sea más placentero, controlado y científico. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.
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Wheat has a specific preference for NO3 - and shows toxicity symptoms under high NH4 + concentrations. Increasing the nitrate supply may alleviate ammonium stress. Nevertheless, the mechanisms underlying the nitrate regulation of wheat root growth to alleviate ammonium toxicity remain unclear. In this study, we integrated physiological and weighted gene co-expression network analysis (WGCNA) to identify the hub genes involved in nitrate alleviation of ammonium toxicity at the wheat seedling stage. Five NH4 +/NO3 - ratio treatments, including 100/0 (Na), 75/25 (Nr1), 50/50 (Nr2), 25/75 (Nr3), and 0/100 (Nn) were tested in this study. The results showed that sole ammonium treatment (Na) increased the lateral root number but reduced root biomass. Increasing the nitrate supply significantly increased the root biomass. Increasing nitrate levels decreased abscisic acid (ABA) content and increased auxin (IAA) content. Furthermore, we identified two modules (blue and turquoise) using transcriptome data that were significantly related to root physiological growth indicators. TraesCS6A02G178000 and TraesCS2B02G056300 were identified as hub genes in the two modules which coded for plastidic ATP/ADP-transporter and WRKY62 transcription factors, respectively. Additionally, network analysis showed that in the blue module, TraesCS6A02G178000 interacts with downregulated genes that coded for indolin-2-one monooxygenase, SRG1, DETOXIFICATION, and wall-associated receptor kinase. In the turquoise module, TraesCS2B02G056300 was highly related to the genes that encoded ERD4, ERF109, CIGR2, and WD40 proteins, and transcription factors including WRKY24, WRKY22, MYB30, and JAMYB, which were all upregulated by increasing nitrate supply. These studies suggest that increasing the nitrate supply could improve root growth and alleviate ammonium toxicity through physiological and molecular regulation networks, including ROS, hormonal crosstalk, and transcription factors.
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B-1 lymphocytes exhibit specialized roles in host defense against multiple pathogens. Despite the fact that CD19+CD93+B220lo/- B cells have been identified as B-1 progenitors, the definition for B-1 progenitors remains to be elucidated as CD19+CD93+B220+ B cells are capable to give rise to B-1 cells. Given that transcription factor Bhlhe41 is highly and preferentially expressed in B-1 cells and regulates B-1a cell development, we generated a transgenic mouse model, Bhlhe41dTomato-Cre , for fate mapping and functional analysis of B-1 cells. Bhlhe41dTomato-Cre mice efficiently traced Bhlhe41 expression, which was mainly restricted to B-1 cells in B-cell lineage. We showed an efficient and specific Cre-mediated DNA recombination in adult B-1 cells and neonatal B-1 progenitors rather than B-2 cells by flow cytometric analysis of Bhlhe41 dTomato-Cre/+ Rosa26 EYFP mice. Treatment of Bhlhe41 dTomato-Cre/+ Rosa26 iDTR mice with diphtheria toxin revealed a robust efficacy of B-1 cell depletion. Interestingly, using Bhlhe41 dTomato-Cre mice, we demonstrated that neonatal B-1 progenitors (CD19+CD93+B220lo/-) expressed Bhlhe41 and were identical to well-defined transitional B-1a progenitors (CD19+CD93+B220lo/-CD5+), which only gave rise to peritoneal B-1a cells. Moreover, we identified a novel population of neonatal splenic CD19hidTomato+B220hiCD43loCD5lo B cells, which differentiated to peritoneal B-1a and B-1b cells. Bhlhe41 deficiency impaired the balance between CD19hidTomato+B220lo/-CD5hi and CD19hidTomato+B220hiCD5lo cells. Hence, we identified neonatal CD19hidTomato+B220hiCD43loCD5lo B cells as novel transitional B-1 progenitors. Bhlhe41 dTomato-Cre/+ mouse can be used for fate mapping and functional studies of B-1 cells in host-immune responses.
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Subgrupos de Linfocitos B , Animales , Antígenos CD19/genética , Antígenos CD19/metabolismo , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , ADN/metabolismo , Toxina Diftérica/metabolismo , Modelos Animales de Enfermedad , Integrasas , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Factores de Transcripción/metabolismoRESUMEN
Toxoplasma gondii, an opportunistic pathogenic protozoan, exhibits a strong predilection to infect the brain, causing severe neurological diseases, such as toxoplasmic encephalitis (TE), in immunocompromised patients. Microglia, the resident immune cells in the brain, is reported to play important roles in regulating the neuroinflammation mediated by T. gondii infection. Here we demonstrated that the tachyzoites of T. gondii RH strain could significantly upregulate the expression levels of microglial M1 phenotype markers including IL-1ß, IL-6, TNF-α, iNOS and IL18 in activated murine BV2 microglia cells, which were regulated by T. gondii rhoptry protein 18 (TgROP18). Moreover, we found that TgROP18 could enhance the expression of M1 phenotype markers in activated murine BV2 microglia cells via activating NF-κB signal pathway. Additionally, TgROP18 was suggested to interact with the host p65 in activated murine BV2 microglia cells and induce the phosphorylation of p65 at S536. In summary, the present study demonstrated that TgROP18 could promote the activated microglia to polarize to M1 phenotype and enhanced the expression of pro-inflammatory factors via activating NF-κB signal pathway, which could contribute to elucidating the mechanism underlying the neuroinflammation mediated by activated microglia in the brain with T. gondii infection.
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Microglía , Toxoplasma , Animales , Biomarcadores , Lipopolisacáridos/metabolismo , Lipopolisacáridos/farmacología , Ratones , Microglía/metabolismo , FN-kappa B/metabolismo , Transducción de SeñalRESUMEN
Human basophils are terminally differentiated granulocytes that are least abundant in the peripheral blood but play important roles in allergic diseases. Studies on human basophils are limited by the high cost on the isolation of human basophils by magnetic-activated cell sorting (MACS) for negative depletion of non-basophils, followed by CD123-based positive selection of basophils. Moreover, such CD123-based purification of basophils may be limited by blocking of the binding of IL-3/anti-CD123 to the surface CD123. Here we identified SSClow CD4- CD127- HLA-DR- CRTH2high as unique markers for the identification of human basophils through stringent flow cytometric analysis of leukocytes from buffy coat. We established an efficient and cost-effective method for isolating human basophils from buffy coat based on positive magnetic selection of CRTH2+ cells followed by flow cytometric sorting of SSClow CD4- CD127- HLA-DR- CRTH2high cells. Approximately 1 to 1.5 million basophils were isolated from one buffy coat with a purity of >97%. Basophils purified by this method were viable and efficiently responded to key regulators of basophils including IL-3 and anti-IgE. This method can be used for purifying human basophils for subsequent functional studies.
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Basófilos , Subunidad alfa del Receptor de Interleucina-3 , Análisis Costo-Beneficio , Antígenos HLA-DR , Humanos , Interleucina-3/metabolismo , Subunidad alfa del Receptor de Interleucina-3/metabolismoRESUMEN
Allergic diseases are caused by dysregulated Th2 immune responses involving multiple effector cells including basophils. Short chain fatty acids (SCFAs), mainly acetate, propionate and butyrate, exert immunomodulatory functions via activation of its receptors GPR41 and GPR43, and inhibition of the histone deacetylases (HDACs) activity. In allergic diseases, SCFAs suppress the activity of mast cells, eosinophils and type 2 innate lymphoid cells (ILC2) but enhance the function of Th2 cells. Here, we aimed to elucidate the function of SCFAs on human basophils. Human basophils were purified from healthy donors by flow cytometric sorting. The surface proteins, apoptosis and degranulation of basophils were analyzed by flow cytometric analysis. The mRNA expression was assayed using real-time PCR. Interleukin 4 (IL-4) and IL-13 were measured by ELISA. Histone acetylation was examined by western blot. GPR41 was expressed by basophils and was enhanced by IL-3. Acetate induced intracellular calcium influx in basophils which was suppressed by blocking GPR41. Propionate and butyrate, but not acetate, induced the expression of CD69 and IL-13. In addition, propionate and butyrate enhanced IgE-mediated basophil degranulation but inhibited basophil survival and IL-4 secretion. Propionate and butyrate induced histone acetylation of basophils and suppression of HDACs activity mimicked the effects of propionate and butyrate on human basophils. Our findings demonstrate that propionate and butyrate may play a complex role in regulating basophil apoptosis, activation and degranulation via inhibiting HDACs activity. The in vivo effects of SCFAs on the regulation of basophil-associated allergic diseases need to be further explored.
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Apoptosis/efectos de los fármacos , Basófilos/efectos de los fármacos , Butiratos/farmacología , Histonas/metabolismo , Interleucina-13/metabolismo , Propionatos/farmacología , Apoptosis/inmunología , Basófilos/inmunología , Basófilos/metabolismo , Células Cultivadas , Eosinófilos/efectos de los fármacos , Eosinófilos/inmunología , Eosinófilos/metabolismo , Ácidos Grasos Volátiles , Expresión Génica/efectos de los fármacos , Expresión Génica/inmunología , Humanos , Hipersensibilidad/inmunología , Hipersensibilidad/metabolismo , Inmunidad Innata/efectos de los fármacos , Inmunidad Innata/inmunología , Mastocitos/efectos de los fármacos , Mastocitos/inmunología , Mastocitos/metabolismo , Células Th2/efectos de los fármacos , Células Th2/inmunología , Células Th2/metabolismoRESUMEN
A series of novel (E)-4-oxo-2-crotonamide derivatives were designed and synthesized to find potent antituberculosis agents. All the target compounds were evaluated for their in vitro activity against Mycobacterium tuberculosis H37Rv(MTB). Results reveal that 4-phenyl moiety at part A and short methyl group at part C were found to be favorable. Most of the derivatives displayed promising activity against MTB with MIC ranging from 0.125 to 4⯵g/mL. Especially, compound IIIa16 was found to have the best activity with MIC of 0.125⯵g/mL against MTB and with MIC in the range of 0.05-0.48⯵g/mL against drug-resistant clinical MTB isolates.
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Antituberculosos/química , Antituberculosos/farmacología , Crotonatos/farmacología , Diseño de Fármacos , Amidas/química , Antituberculosos/síntesis química , Crotonatos/química , Evaluación Preclínica de Medicamentos , Pruebas de Sensibilidad Microbiana , Mycobacterium tuberculosis/efectos de los fármacos , Relación Estructura-ActividadRESUMEN
We studied the pharmacokinetics, biodistribution and metabolism of phospho-sulindac (PS), a novel agent efficacious in the treatment of dry eye, formulated in nanoparticles (PS-NPs) following its topical administration to the eye of New Zealand White rabbits. The nanoparticles were spherical with effective diameterâ¯=â¯108.9⯱â¯41.7â¯nm, zeta potentialâ¯=â¯-21.70⯱â¯3.78â¯mV, drug loadingâ¯=â¯7%, and entrapment efficiencyâ¯=â¯46.4%. Of the total PS delivered topically to the eye, >95% was retained in the anterior segment, predominantly in the cornea (Cmaxâ¯=â¯101.3⯵M; Tmaxâ¯=â¯1â¯h; T1/2â¯=â¯2.6â¯h; area AUC0-16hâ¯=â¯164.4⯵M·h) and conjunctiva (Cmaxâ¯=â¯89.4⯵M; Tmaxâ¯=â¯0.25â¯h; T1/2â¯=â¯3.1â¯h; AUC0-16hâ¯=â¯63.5⯵M·h), the tissues most affected by dry eye disease. No PS or its metabolites were detected in the systemic circulation. PS was metabolized to PS sulfide and PS sulfone; all three molecules were hydrolyzed to sulindac, which was converted to sulindac sulfide and sulindac sulfone. A solution formulation of PS provided lower PS levels in ocular tissues but higher levels of PS metabolites, compared to PS-NPs. Therefore, NPs represent an effective formulation for the topical ocular administration of PS for anterior segment diseases, such as dry eye disease.
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Sistemas de Liberación de Medicamentos , Ojo/metabolismo , Nanopartículas/administración & dosificación , Compuestos Organofosforados/administración & dosificación , Sulindac/análogos & derivados , Administración Intravenosa , Administración Tópica , Animales , Masculino , Nanopartículas/química , Compuestos Organofosforados/sangre , Compuestos Organofosforados/química , Compuestos Organofosforados/farmacocinética , Conejos , Sulindac/administración & dosificación , Sulindac/sangre , Sulindac/química , Sulindac/farmacocinética , Distribución TisularRESUMEN
The manipulation of bile acid (BA) homeostasis by blocking the ileal apical Na+-dependent bile salt transporter (ASBT/SLC10A2) may have therapeutic effects in nonalcoholic fatty liver disease. We developed a novel ASBT inhibitor, an N-(3,4-o-dichlorophenyl)-2-(3-trifluoromethoxy) benzamide derivative referred to as IMB17-15, and investigated its therapeutic effects and the molecular mechanisms underlying the effects. Syrian golden hamsters were challenged with high-fat diet (HFD) to induce NAFLD and were subsequently administered 400 mg/kg IMB17-15 by gavage daily for 21 days. Serum, liver, and fecal samples were collected for further analysis. Plasma concentration-time profiles of IMB17-15 were also constructed. The human hepatocyte cell line HL-7702 was treated with Oleic acid (OA) with or without IMB17-15. Western blotting and real-time PCR were used to study the molecular mechanisms of IMB17-15. We found that IMB17-15 inhibited ASBT and subsequently suppressed ileal farnesoid X receptor (FXR) and FXR-activated fibroblast growth factor15/19 (FGF15/19) expression, which reduced the hepatic phosphorylated extracellular regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK) levels and upregulated the cholesterol 7α-hydroxylase (CYP7A1) activity. Additionally, IMB17-15 stimulated adenosine monophosphate (AMP)-activated protein kinase (AMPKα) phosphorylation and enhanced peroxisome proliferator activated receptor α (PPARα) expression and thus promoted triglyceride (TG) oxidation and high-density lipoprotein cholesterol (HDL-c) metabolism through an ASBT-independent mechanism. In conclusion, a novel ASBT inhibitor known as IMB17-15 protected hamsters against HFD-induced NFALD by manipulating BA and lipid homeostasis. IMB17-15 also reduced lipid deposition in human hepatic cell lines, indicating that it may be useful as a therapy for NAFLD patients.
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Benzamidas/uso terapéutico , Enfermedad del Hígado Graso no Alcohólico/prevención & control , Transportadores de Anión Orgánico Sodio-Dependiente/antagonistas & inhibidores , Sulfonamidas/uso terapéutico , Simportadores/antagonistas & inhibidores , Animales , Benzamidas/farmacocinética , Benzamidas/toxicidad , Línea Celular , Citocinas/metabolismo , Dieta Alta en Grasa , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Hígado/patología , Masculino , Mesocricetus , Ratones , Enfermedad del Hígado Graso no Alcohólico/genética , Enfermedad del Hígado Graso no Alcohólico/patología , Sulfonamidas/farmacocinética , Sulfonamidas/toxicidadRESUMEN
The apical sodium--dependent bile acid transporter (ASBT) is the main transporter to promote re-absorption of bile acids from the intestinal tract into the enterohepatic circulation. Inhibition of ASBT could increase the excretion of bile acids, thus increasing bile acid synthesis and consequently cholesterol consumption. Therefore, ASBT is an attractive target for developing new cholesterol-lowering drugs. In this report, a series of 1-(2,4-bifluorophenyl)-7-dialkylamino-1,8-naphthyridine-3-carboxamides were designed as inhibitors of ASBT. Most of them demonstrated potency against ASBT transport of bile acids. In particular, compound 4a1 was found to have the best activity, resulting in 80.1% inhibition of ASBT at 10 µmol/L.
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Mesoporous silica-encapsulated gold nanorods (GNRs@mSiO(2)) have great potential both in photothermal therapy and drug delivery. In this paper, we firstly developed GNRs@mSiO(2) as a synergistic therapy tool for delivery heat and drug to the tumorigenic region. We studied the ablation of tumor both in vitro and in vivo by the combination of photothermal therapy and chemotherapy using doxorubicin (DOX)-loaded GNRs@mSiO(2). Significantly greater cell killing was observed when A549 cells incubated with DOX-loaded GNRs@mSiO(2) were irradiated with near-infrared (NIR) illumination, attributable to both GNRs@mSiO(2)-mediated photothermal ablation and cytotoxicity of light-triggered DOX release. We then performed in vivo therapy studies and observed a promising tumor treatment. Compared with chemotherapy or photothermal treatment alone, the combined treatment showed a synergistic effect, resulting in higher therapeutic efficacy. Furthermore, the lower systematic toxicity of GNRs@mSiO(2) has been validated.
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Oro/química , Nanotubos , Neoplasias Experimentales/terapia , Animales , Antineoplásicos/uso terapéutico , Línea Celular Tumoral , Terapia Combinada , Sistemas de Liberación de Medicamentos , Humanos , Hipertermia Inducida , Masculino , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica de TransmisiónRESUMEN
In this paper, a facile solvothermal synthesis method was developed to prepare monodisperse magnetites anchored onto multi-walled carbon nanotubes (MWNTs). The pristine MWNTs were treated with a mixture of concentrated sulfuric and nitric acids. The oxidized MWNTs (o-MWNTs) had abundant carboxylic groups on the surface, which have a strong ability to chelate metal ions like Fe3+. The obtained MWNTs-Fe3O4 nanomaterials allowed π-π stacking of fluorescein isothiocyanate (FITC) to monitor inside living cancer cells by fluorescence imaging. Cells labeled with MWNTs-Fe3O4 nanomaterials could be efficiently manipulated by a magnetic field due to the large magnetic moment of the iron oxide domain in the nanocomposites. The MWNTs-Fe3O4 nanomaterials have been demonstrated to be effective for in vivo photothermal treatment of tumors using mice implanted with human U87 tumors as a model.
RESUMEN
Phage-displayed TGN peptide-decorated polymeric micelle-like polyplexes based on pegylated poly(2-(dimethylamino) ethyl methacrylate) (PEG-PDMAEMA) were prepared for efficient brain-targeted gene delivery. The diblock copolymers Methoxy-PEG-PDMAEMA and Maleimide-PEG-PDMAEMA were synthesized by the atom transfer radical polymerization method. The TGN ligand, a 12-amino acid peptide that could facilitate blood-brain barrier (BBB) targeting, was conjugated to the PEG terminus of the copolymer via a maleimide-mediated covalent binding procedure. TGN-PEG-PDMAEMA was complexed with plasmid DNA to yield polyplexes. The physiochemical properties of the polyplexes, such as morphology, particle size, zeta potential, cytotoxicity and DNA complex formation ability, were studied prior to the successful in vitro and in vivo transfection. The TGN-PEG-PDMAEMA/DNA polyplexes maintained their stable nano-size, were characterized by good condensation capacity and low toxicity and even provided higher cellular uptake than the unmodified polyplexes (PEG-PDMAEMA/DNA polyplexes). Confocal microscopy studies showed that the DNA of TGN-PEG-PDMAEMA/DNA polyplexes entered into the nuclei through the endosome/lysosome pathway. The transfection efficiency of TGN-modified polyplexes was higher than that of unmodified polyplexes both in vitro and in vivo. The results obtained from frozen sections indicated the widespread expression of an exogenous gene in the mouse brain after intravenous injection. Therefore, the results demonstrate that the TGN-decorated PEG-PDMAEMA developed in this study could be utilized as a potential vehicle for gene delivery to the brain.