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OBJECTIVES: Disseminated fibrin-rich microthrombi have been reported in patients who died from COVID-19. Our objective is to determine whether the fibrin clot structure and function differ between critically ill patients with or without COVID-19 and to correlate the structure with clinical coagulation biomarkers. DESIGN: A cross-sectional observational study. Platelet poor plasma was used to analyze fibrin clot structure; the functional implications were determined by quantifying clot turbidity and porosity. SETTING: ICU at an academic medical center and an academic laboratory. PATIENTS: Patients admitted from July 1 to August 1, 2020, to the ICU with severe acute respiratory syndrome coronavirus 2 infection confirmed by reverse transcription-polymerase chain reaction or patients admitted to the ICU with sepsis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood was collected from 36 patients including 26 ICU patients with COVID-19 and 10 ICU patients with sepsis but without COVID-19 at a median of 11 days after ICU admission (interquartile range, 3-16). The cohorts were similar in age, gender, body mass index, comorbidities, Sequential Organ Failure Assessment (SOFA) score, and mortality. More patients with COVID-19 (100% vs 70%; p = 0.003) required anticoagulation. Ex vivo fibrin clots formed from patients with COVID-19 appeared to be denser and to have smaller pores than those from patients with sepsis but without COVID-19 (percent area of fluorescent fibrin 48.1% [SD, 16%] vs 24.9% [SD, 18.8%]; p = 0.049). The turbidity and flow-through assays corroborated these data; fibrin clots had a higher maximum turbidity in patients with COVID-19 compared with patients without COVID-19 (0.168 vs 0.089 OD units; p = 0.003), and it took longer for buffer to flow through these clots (216 vs 103 min; p = 0.003). In patients with COVID-19, d-dimer levels were positively correlated with percent area of fluorescent fibrin (ρ = 0.714, p = 0.047). Denser clots (assessed by turbidity and thromboelastography) and higher SOFA scores were independently associated with delayed clot lysis. CONCLUSIONS: We found aberrant fibrin clot structure and function in critically ill patients with COVID-19. These findings may contribute to the poor outcomes observed in COVID-19 patients with widespread fibrin deposition.
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COVID-19 , Sepsis , Tromboembolia , Trombosis , Enfermedad Crítica , Estudios Transversales , Fibrina , Fibrinólisis , HumanosRESUMEN
Silver nanoparticles (AgNPs) are considered as emerging contaminants because of their high toxicity and increasing environmental impact. Removal of discharged AgNPs from water is crucial for mitigating the health and environmental risks. However, developing facile, economical, and environment-friendly approaches remains challenging. Herein, an Fe3O4-Mg(OH)2 nanocomposite, as a novel magnetic scavenger for AgNPs, was prepared by loading Fe3O4 nanoparticles on Mg(OH)2 nanoplates in a one-pot synthesis. Batch removal experiments revealed that the maximum removal capacities for the two model AgNPs (citrate- or polyvinylpyrrolidone-coated AgNPs) were 476 and 442 mg/g, respectively, corresponding to partition coefficients 8.03 and 4.89 mg/g/µM. Removal feasibilities over a wide pH range of 5-11 and in real water matrices and scavenger reusability with five cycles were also confirmed. Both Fe3O4 and Mg(OH)2 components contributed to the removal; however, their nanocomposites exhibited an enhanced performance because of the high specific surface area and pore volume. Chemical adsorption and electrostatic attraction between the coatings on the AgNPs and the two components in the nanocomposite was considered to be responsible for the removal. Overall, the facile synthesis, convenient magnetic separation, and high removal performance highlight the great potential of the Fe3O4-Mg(OH)2 nanocomposite for practical applications.
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Nanopartículas del Metal , Nanocompuestos , Adsorción , Plata , AguaRESUMEN
The widespread use of silver nanoparticles (AgNPs) inevitably leads to the environmental release of AgNPs. The released AgNPs can pose ecological risks because of their specific toxicity. However, they can also be used as secondary sources of silver metal. Herein, hierarchical mesoporous calcite (HMC) was prepared and used to remove and recover AgNPs from an aqueous solution. The batch experiments show that the HMC has high removal percentages for polyvinylpyrrolidone- and poly (vinyl alcohol)-coated AgNPs (PVP- and PVA-AgNPs) over a wide pH range of 6-10. The adsorption isotherms indicate that the maximum removal capacities are 55 and 19 mg g-1 for PVP-AgNPs and PVA-AgNPs, respectively, corresponding to partition coefficients (PCs) of 0.55 and 0.77 mg g-1 µM-1. Furthermore, the removal performance is also not impaired by coexisting anions, such as Cl-, NO3-, SO42-, and CO32-. Their removal mechanisms can be ascribed to the electrostatic attraction and chemical adsorption between the HMC and polymer-coated AgNPs. Calcium ions on the HMC surface serve as active sites for coordination with the oxygen-bearing functional groups of AgNP coatings. Moreover, the AgNPs adsorbed onto HMC show high catalytic activity and good reusability for the reduction of the organic pollutant 4-nitrophenol. This work may pave the way not only to remove metal nanopollutants from waters but also to convert them into functional materials.
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Nanopartículas del Metal , Plata , Adsorción , Carbonato de Calcio , PolímerosRESUMEN
We have previously shown that ritonavir (RTV), a highly active anti-retroviral therapy (HAART) drug, can cause endothelial dysfunction through oxidative stress. Several antioxidants including ginsenoside Rb1, a compound with antioxidant effect, can effectively block this side effect of RTV in endothelial cells. In the current study, we explored a mechanism by which ginsenoside Rb1 could protect these cells via binding of estrogen receptors (ERs). We found that several human endothelial cell lines differentially expressed ER-ß and had very low levels of ER-α. RTV treatment significantly increased the production of reactive oxygen species (ROS) and decreased the expression of endothelial nitric oxidase synthase (eNOS) and superoxide dismutase (SOD) in HUVECs, while Rb1 effectively blocked these effects of RTV. These effects of Rb1 were effectively inhibited by silencing ER-ß, indicating that ginsenoside Rb1 requires ER-ß for its antioxidant activity in inhibiting the deleterious effect of RTV in human endothelial cells. Furthermore, Rb1 specifically activated ER-ß transactivation activity by ER-ß luciferase reporter assay. Rb1 competitively bound to ER-ß, which was determined by the high sensitive fluorescent polarization assay.
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Células Endoteliales/metabolismo , Receptor beta de Estrógeno/genética , Ginsenósidos/farmacología , Óxido Nítrico Sintasa de Tipo III/metabolismo , Ritonavir/efectos adversos , Superóxido Dismutasa/metabolismo , Línea Celular , Regulación hacia Abajo , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Receptor beta de Estrógeno/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Humanos , Óxido Nítrico Sintasa de Tipo III/genética , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/genética , Activación Transcripcional , Regulación hacia ArribaRESUMEN
BACKGROUND Entacapone (ENT), a clinical drug for the treatment of Parkinson's disease, has been shown to have antioxidant effects, but little is known about its antioxidant mechanisms. The objective of the current study was to determine the antioxidant activity of ENT against different species of oxidants and compared it with that of vitamin C and vitamin E. We also determined the effect of ENT on oxidative stress-induced cell death in human umbilical vein endothelial cells (HUVECs). MATERIAL AND METHODS The total antioxidant activities of ENT, vitamin C and vitamin E were determined with a standard DPPH-scavenging assay. Specific assays to determine ENT's scavenging activity on hypochlorous acid (HOCl), peroxynitrite (ONOO-), and hydrogen peroxide (H2O2), and the chelating effect on Fe(II) were used. H2O2-induced cell death in HUVECs was determined with the MTT assay. RESULTS ENT (10 and 20 µM) scavenged 60% and 83% of DPPH activity, respectively. These percentages were greater than those resulting from using the same concentrations of vitamin C and vitamin E. ENT's HOCl-scavenging activity was concentration-dependent and 8 to 20 times stronger than those of vitamin C and vitamin E. ENT's ONOO--scavenging activity was 8% to 30% stronger than that of vitamin C. However, ENT, vitamin C, and vitamin E were not able to directly scavenge H2O2, and did not show any chelating effect on Fe(II). Importantly ENT, but not vitamin C or vitamin E, inhibited H2O2-induced cell death in HUVECs. CONCLUSIONS ENT is an antioxidant that can scavenge toxic HOCl and ONOO- species and inhibit oxidative stress-induced cell death more effectively than vitamin C and vitamin E. ENT may have new clinical applications as an antioxidant in the treatment of ROS-induced diseases including cardiovascular disease, cancer, and neurodegenerative diseases.
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Antioxidantes/farmacología , Ácido Ascórbico/farmacología , Catecoles/farmacología , Depuradores de Radicales Libres/farmacología , Ácido Hipocloroso/farmacología , Nitrilos/farmacología , Estrés Oxidativo/efectos de los fármacos , Ácido Peroxinitroso/farmacología , Vitamina E/farmacología , Muerte Celular/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/farmacología , Quelantes del Hierro/farmacología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Uric acid is the final oxidation product of purine metabolism in humans. Xanthine oxidoreductase (XOR) catalyzes oxidative hydroxylation of hypoxanthine to xanthine to uric acid, accompanying the production of reactive oxygen species (ROS). Uric acid usually forms ions and salts known as urates and acid urates in serum. Clinically, overproduction or under-excretion of uric acid results in the elevated level of serum uric acid (SUA), termed hyperuricemia, which has long been established as the major etiologic factor in gout. Accordingly, urate-lowering drugs such as allopurinol, an XOR-inhibitor, are extensively used for the treatment of gout. In recent years, the prevalence of hyperuricemia has significantly increased and more clinical investigations have confirmed that hyperuricemia is an independent risk factor for cardiovascular disease, hypertension, diabetes, and many other diseases. Urate-lowering therapy may also play a critical role in the management of these diseases. However, current XOR-inhibitor drugs such as allopurinol and febuxostat may have significant adverse effects. Therefore, there has been great effort to develop new XOR-inhibitor drugs with less or no toxicity for the long-term treatment or prevention of these hyperuricemia-related diseases. In this review, we discuss the mechanism of uric acid homeostasis and alterations, updated prevalence, therapeutic outcomes, and molecular pathophysiology of hyperuricemia-related diseases. We also summarize current discoveries in the development of new XOR inhibitors.
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Inhibidores Enzimáticos/farmacología , Hiperuricemia/enzimología , Hiperuricemia/terapia , Xantina Deshidrogenasa/antagonistas & inhibidores , Xantina Deshidrogenasa/metabolismo , Alopurinol/farmacología , Animales , Inhibidores Enzimáticos/uso terapéutico , Febuxostat/farmacología , Humanos , Especies Reactivas de Oxígeno/metabolismo , Factores de RiesgoRESUMEN
Cyclophilin A (CypA) was shown to be upregulated in human cholangiocarcinoma (CCA) tissues. Suppression of intracellular CypA (inCypA) significantly reduces cell proliferation in vitro and tumor growth in nude mice. In the present study, the effect and potential mechanism of secreted CypA (sCypA) on cell proliferation of CCA cell lines were further investigated. CCA cells were treated with sCypA-containing conditioned media (CM) or with purified recombinant human CypA (rhCypA). Cell proliferation, cell cycle, ERK1/2, p38 MAPK, NF-κB, and STAT3 activities were examined by MTS assay, flow cytometry, and Western blot. sCypA was detected in CM from MMNK1 (an immortalized human cholangiocyte cell line) and six CCA cell lines. The sCypA levels corresponded to the inCypA levels indicating the intracellular origin of sCypA. Both sCypA-containing CM and rhCypA significantly increased proliferation of CCA cells. CD147 depletion by shRNA-knockdown or neutralizing with a CD147-monoclonal antibody significantly reduced sCypA-, and rhCypA-mediated cell proliferation. Upon rhCypA treatment, ERK1/2 was rapidly phosphorylated; whereas neutralizing CD147 inhibited ERK1/2 phosphorylation. Cell cycle analysis showed a significant increase in S phase and decrease in G1 population in rhCypA-treated cells. The expression levels of cyclin D1 and phosphorylated-retinoblastoma protein in the rhCypA-treated cells were increased compared with those in the non-treated control cells. p38 MAPK pathway was shown to be suppressed in siCypA-treated cells. In summary, CypA is secreted from CCA cells and enhances cell proliferation in an autocrine/paracrine manner, at least via direct binding with CD147, which may activate the ERK1/2 and p38 MAPK signaling pathways.
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Basigina/biosíntesis , Neoplasias de los Conductos Biliares/genética , Colangiocarcinoma/genética , Ciclofilina A/genética , Proteínas Quinasas p38 Activadas por Mitógenos/biosíntesis , Animales , Basigina/genética , Neoplasias de los Conductos Biliares/patología , Línea Celular Tumoral , Proliferación Celular/genética , Colangiocarcinoma/patología , Medios de Cultivo Condicionados , Ciclofilina A/biosíntesis , Ciclofilina A/metabolismo , Puntos de Control de la Fase G1 del Ciclo Celular/genética , Regulación Neoplásica de la Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , Sistema de Señalización de MAP Quinasas/genética , Ratones , ARN Mensajero/biosíntesis , ARN Interferente Pequeño/genética , Proteínas Quinasas p38 Activadas por Mitógenos/genéticaRESUMEN
Pancreatic ductal adenocarcinoma (PDAC) remains a lethal malignancy, largely due to the paucity of reliable biomarkers for early detection and therapeutic targeting. Existing blood protein biomarkers for PDAC often suffer from replicability issues, arising from inherent limitations such as unmeasured confounding factors in conventional epidemiologic study designs. To circumvent these limitations, we use genetic instruments to identify proteins with genetically predicted levels to be associated with PDAC risk. Leveraging genome and plasma proteome data from the INTERVAL study, we established and validated models to predict protein levels using genetic variants. By examining 8,275 PDAC cases and 6,723 controls, we identified 40 associated proteins, of which 16 are novel. Functionally validating these candidates by focusing on 2 selected novel protein-encoding genes, GOLM1 and B4GALT1, we demonstrated their pivotal roles in driving PDAC cell proliferation, migration, and invasion. Furthermore, we also identified potential drug repurposing opportunities for treating PDAC. SIGNIFICANCE: PDAC is a notoriously difficult-to-treat malignancy, and our limited understanding of causal protein markers hampers progress in developing effective early detection strategies and treatments. Our study identifies novel causal proteins using genetic instruments and subsequently functionally validates selected novel proteins. This dual approach enhances our understanding of PDAC etiology and potentially opens new avenues for therapeutic interventions.
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Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Humanos , Proteoma , Carcinoma Ductal Pancreático/diagnóstico , Carcinoma Ductal Pancreático/genética , Glicosiltransferasas , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/genética , Biomarcadores , Proteínas de la MembranaRESUMEN
ß-thymosins, including thymosin ß4 (Tß4), Tß10, and Tß15, are a family of highly conserved 5 kDa peptides. They are involved not only in normal cell migration, but also in tumor metastasis. However, the molecular mechanisms of ß-thymosins to regulate cell migration and other functions are not fully understood. Recently, this important area is under active investigation worldwide. Many new discoveries have been made from molecular biology and cell culture models as well as animal models and human diseases. This timely review provides the most updated information about functional roles and molecular mechanisms of ß-thymosins in normal tissues and disease conditions.
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Movimiento Celular/fisiología , Neoplasias/metabolismo , Neoplasias/patología , Timosina/metabolismo , Actinas/metabolismo , Animales , Transición Epitelial-Mesenquimal , Humanos , Metástasis de la Neoplasia , Timosina/químicaRESUMEN
Sieve and take: A biomimetic strategy was designed to fabricate two-dimensional silica sieve plates (SSP) by use of catanionic surfactants as composite template and L-tartrate with hydroxyl and carboxyl groups as regulator. Tartrate was found to combine two capabilities in the formation of SSP structures: the connection of adjacent silica structures through H bonding and the separation of adjacent structures through electrostatic repulsion.
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BACKGROUND: Thymosin ß10 (Tß10) expression is associated with malignant phenotypes in many cancers. However, the role and mechanisms of Tß10 in liver fluke-associated cholangiocarcinoma (CCA) are not fully understood. In this study, we investigated the expression of Tß10 in CCA tumor tissues and cell lines as well as molecular mechanisms of Tß10 in tumor metastasis of CCA cell lines. METHODS: Tß10 expression was determined by real time RT-PCR or immunocytochemistry. Tß10 silence or overexpression in CCA cells was achieved using gene delivery techniques. Cell migration was assessed using modified Boyden chamber and wound healing assay. The effect of silencing Tß10 on CCA tumor metastasis was determined in nude mice. Phosphorylation of ERK1/2 and the expression of EGR1, Snail and matrix metalloproteinases (MMPs) were studied. RESULTS: Ten pairs of CCA tissues (primary and metastatic tumors) and 5 CCA cell lines were studied. With real time RT-PCR and immunostaining analysis, Tß10 was highly expressed in primary tumors of CCA; while it was relatively low in the metastatic tumors. Five CCA cell lines showed differential expression levels of Tß10. Silence of Tß10 significantly increased cell migration, invasion and wound healing of CCA cells in vitro; reversely, overexpression of Tß10 reduced cell migration compared with control cells (P<0.05). In addition, silence of Tß10 in CCA cells increased liver metastasis in a nude mouse model of CCA implantation into the spleen. Furthermore, silence of Tß10 activated ERK1/2 and increased the expression of Snail and MMPs in CCA cell lines. Ras-GTPase inhibitor, FPT inhibitor III, effectively blocked Tß10 silence-associated ERK1/2 activation, Snail expression and cell migration. CONCLUSIONS: Low expression of Tß10 is associated with metastatic phenotype of CCA in vitro and in vivo, which may be mediated by the activation of Ras, ERK1/2 and upregulation of Snail and MMPs. This study suggests a new molecular pathway of CCA pathogenesis and a novel strategy to treat or prevent CCA metastasis.
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Movimiento Celular/genética , Colangiocarcinoma/genética , Colangiocarcinoma/patología , Timosina/genética , Animales , Línea Celular Tumoral , Colangiocarcinoma/etiología , Colangiocarcinoma/metabolismo , Modelos Animales de Enfermedad , Fasciola hepatica , Fascioliasis/complicaciones , Expresión Génica , Silenciador del Gen , Humanos , Ratones , Ratones Desnudos , Metástasis de la Neoplasia , Interferencia de ARN , Timosina/metabolismoRESUMEN
Recent advances in human genomics and biotechnologies have profound impacts on medical research and clinical practice. Individual genomic information, including DNA sequences and gene expression profiles, can be used for prediction, prevention, diagnosis, and treatment for many complex diseases. Personalized medicine attempts to tailor medical care to individual patients by incorporating their genomic information. In a case of pancreatic cancer, the fourth leading cause of cancer death in the United States, alteration in many genes as well as molecular profiles in blood, pancreas tissue, and pancreas juice has recently been discovered to be closely associated with tumorigenesis or prognosis of the cancer. This review aims to summarize recent advances of important genes, proteins, and microRNAs that play a critical role in the pathogenesis of pancreatic cancer, and to provide implications for personalized medicine in pancreatic cancer.
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Regulación Neoplásica de la Expresión Génica/genética , Genómica/métodos , MicroARNs/metabolismo , Proteínas de Neoplasias/metabolismo , Oncogenes/genética , Neoplasias Pancreáticas/genética , Medicina de Precisión/métodos , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/metabolismo , Genómica/tendencias , Humanos , MicroARNs/genética , Neoplasias Pancreáticas/metabolismo , Medicina de Precisión/tendenciasRESUMEN
A major challenge for radioactive wastewater treatment and associated environmental remediation is how to simultaneously remove cationic and anionic radionuclides. Herein, a series of Mn3O4@polyaniline (Mn3O4@PANI) nanocomposites were successfully prepared and used to remove U(VI) and I- from aqueous solution, two highly concomitant species in nuclear pollution settings. Batch adsorption experiments reveal that the component Mn3O4 is predominantly responsible for U(VI) removal, but PANI for I-. The nanocomposite with 24.2 wt% Mn3O4 possesses high removal percentages (> 85%) either for U(VI) or I- over a wide pH range, fast removal kinetics, and excellent adsorption selectivity at high concentrations of competing ions. Benefiting from the contributions of the two components and the high adsorption affinities, the nanocomposite achieves the simultaneous removal to coexisting U(VI) and I-, with a maximum adsorption capacity 102.6 mg/g for U(VI) and 126.1 mg/g for I-. X-ray photoelectron spectroscopy (XPS) results reveal that the U(VI) adsorption occurs via coordination bonding with Mn-O, -NH- , and =N- groups in the nanocomposite, whereas I- adsorption proceeds mainly through I anionic species exchange with Cl- and interactions with π-bonds in PANI, as well as the electrostatic attraction onto Mn3O4. Considering the excellent performance and multiple active sites, the Mn3O4@PANI nanocomposite is promising to remove practical radioactive U(VI) and I-.
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Nanocompuestos , Uranio , Yoduros , Uranio/análisis , Dominio Catalítico , Cationes , Nanocompuestos/química , Adsorción , CinéticaRESUMEN
Organic dyes and heavy metals often coexist in industrial effluents, and their simultaneous removal is a grand challenge. Herein, a hydrochar and MgAl layered double hydroxide (HC-MgAlLDH) nanocomposite was prepared via a facile one-step hydrothermal route, and applied to remove anionic Congo red (CR), cationic Methylene blue (MB) and Pb(II) from aqueous solutions. The nanocomposite was formed by interweaving amorphous HC and crystalline MgAlLDH nanoplates and possessed more functional groups, lower zeta potential and larger specific surface area than uncomposited MgAlLDH. Batch removal experiments showed that the components HC and LDH dominated the CR and MB removals, respectively, whereas Pb(II) removal was conjointly controlled by the two components. The maximum Langmuir removal capacities of the nanocomposite to sole CR, MB, or Pb(II) were 348.78, 256.54 or 33.55 mg/g. In binary and ternary systems, the removal capacities of CR and MB only slightly decreased, while the capacity of Pb(II) increased by 41.13-88.61%. The increase was related to the coordination of Pb(II) with the sulfur-containing groups in dyes and the precipitation of PbSO4. Therefore, the simultaneous removal of CR, MB and Pb(II) was involved in a synergistic effect, including electrostatic adsorption, π-π interaction, coordination and precipitation. The present work shows that the HC-MgAlLDH nanocomposite has great potential for wastewater integrative treatment.
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Pancreatic ductal adenocarcinoma (PDAC) remains an extremely aggressive disease characterized by rapidly acquired multi-drug resistance, including to first-line chemotherapeutic agent gemcitabine. Autophagy is a process that is often exploited by cancer and is one of several intrinsic factors associated with resistance to gemcitabine. We have previously found that miR-198 acts as a tumor suppressor in PDAC through the targeting of factors including Valosin-containing protein (VCP). VCP has been reported to play an important role in autophagic flux. In this study, we investigated whether the repression of VCP through miR-198 administration disrupts the autophagy process and sensitizes PDAC cells to gemcitabine treatment in vitro. Moreover, we used LGA-PEI (LPNP) nanoparticles to effectively administer miR-198 to tumors in vivo, inducing tumor sensitization to gemcitabine and leading to a significant reduction in tumor burden and metastases and a concomitant downregulation of VCP expression and autophagy maturation. Our results indicate a potential therapeutic strategy for targeting gemcitabine resistant PDAC and establishes the use of LPNPs for effective therapeutic delivery of nucleic acids in vitro and in vivo.
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Despite substantial advances in targeting mutant KRAS, tumor resistance to KRAS inhibitors (KRASi) remains a major barrier to progress. Here, we report proteostasis reprogramming as a key convergence point of multiple KRASi-resistance mechanisms. Inactivation of oncogenic KRAS down-regulated both the heat shock response and the inositol-requiring enzyme 1α (IRE1α) branch of the unfolded protein response, causing severe proteostasis disturbances. However, IRE1α was selectively reactivated in an ER stress-independent manner in acquired KRASi-resistant tumors, restoring proteostasis. Oncogenic KRAS promoted IRE1α protein stability through extracellular signal-regulated kinase (ERK)-dependent phosphorylation of IRE1α, leading to IRE1α disassociation from 3-hydroxy-3-methylglutaryl reductase degradation (HRD1) E3-ligase. In KRASi-resistant tumors, both reactivated ERK and hyperactivated AKT restored IRE1α phosphorylation and stability. Suppression of IRE1α overcame resistance to KRASi. This study reveals a druggable mechanism that leads to proteostasis reprogramming and facilitates KRASi resistance.
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Antineoplásicos , Resistencia a Antineoplásicos , Endorribonucleasas , Inhibidores Enzimáticos , Quinasas MAP Reguladas por Señal Extracelular , Factores de Transcripción del Choque Térmico , Neoplasias , Proteostasis , Proteínas Proto-Oncogénicas p21(ras) , Humanos , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Proteínas Serina-Treonina Quinasas , Proteínas Proto-Oncogénicas p21(ras)/antagonistas & inhibidores , Proteínas Proto-Oncogénicas p21(ras)/genética , Inhibidores Enzimáticos/farmacología , Antineoplásicos/farmacología , Factores de Transcripción del Choque Térmico/metabolismoRESUMEN
BACKGROUND: Several HIV protease mutations, which are resistant to clinical HIV protease inhibitors (PIs), have been identified. There is a great need for second-generation PIs with different chemical structures and/or with an alternative mode of inhibition. Ginkgolic acid is a natural herbal substance and a major component of the lipid fraction in the nutshells of the Ginkgo biloba tree. The objective of this study was to determine whether ginkgolic acid could inhibit HIV protease activity in a cell free system and HIV infection in human cells. MATERIAL/METHODS: Purified ginkgolic acid and recombinant HIV-1 HXB2 KIIA protease were used for the HIV protease activity assay. Human peripheral blood mononuclear cells (PBMCs) were used for HIV infection (HIV-1SF162 virus), determined by a p24gag ELISA. Cytotoxicity was also determined. RESULTS: Ginkgolic acid (31.2 µg/ml) inhibited HIV protease activity by 60%, compared with the negative control, and the effect was concentration-dependent. In addition, ginkgolic acid treatment (50 and 100 µg/ml) effectively inhibited the HIV infection at day 7 in a concentration-dependent manner. Ginkgolic acid at a concentration of up to 150 µg/ml demonstrated very limited cytotoxicity. CONCLUSIONS: Ginkgolic acid effectively inhibits HIV protease activity in a cell free system and HIV infection in PBMCs without significant cytotoxicity. Ginkgolic acid may inhibit HIV protease through different mechanisms than current FDA-approved HIV PI drugs. These properties of ginkgolic acid make it a promising therapy for HIV infection, especially as the clinical problem of viral resistance to HIV PIs continues to grow.
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Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , Inhibidores de la Proteasa del VIH/farmacología , Inhibidores de la Proteasa del VIH/uso terapéutico , Proteasa del VIH/metabolismo , Salicilatos/farmacología , Salicilatos/uso terapéutico , Muerte Celular/efectos de los fármacos , Sistema Libre de Células , Relación Dosis-Respuesta a Droga , Ginkgólidos/química , Ginkgólidos/farmacología , Ginkgólidos/uso terapéutico , Proteasa del VIH/efectos de los fármacos , Inhibidores de la Proteasa del VIH/química , Humanos , Células Jurkat , Lactonas/química , Lactonas/farmacología , Lactonas/uso terapéutico , Salicilatos/químicaRESUMEN
Most cancers harbor a diverse collection of cell types including a typically heterogeneous cancer cell fraction. To reconstruct cell-intrinsic and heterotypic interactions driving tumor progression, we combine the XDec deconvolution method with cell-type-specific gene expression correlation analysis into the XDec-CHI method. XDec-CHI identifies intra- and inter-cellular pathways using correlation and places them in the context of specific tumor subtypes, as defined by the state of constituent cancer cells. We make the method web-accessible for analysis of publicly accessible pancreatic ductal adenocarcinoma, breast, head and neck, glioblastoma, and glioma tumors. We apply the method to TCGA and ICGC datasets to identify immune-suppressive interactions within PDAC tumors that are relevant for immunotherapies targeting PD-L1. Subtype-specific interactions derived from correlative analyses validated in co-culture experiments suggest PDAC subtypes have distinct therapeutic weaknesses, with Basal-like and MSLN-high Classical B tumors most likely to respond to therapies targeting PD-L1.
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A major objective in the exploration of Mars is to test the hypothesis that the planet has ever hosted life. Biogenic compounds, especially biominerals, are believed to serve as biomarkers in Raman-assisted remote sensing missions. However, the prerequisite for the development of these minerals as biomarkers is the uniqueness of their biogenesis. Herein, tetragonal bipyramidal weddellite, a type of calcium oxalate, is successfully achieved by UV-photolyzing pyruvic acid (PA). The as-prepared products are identified and characterized by micro-Raman spectroscopy and field emission scanning electron microscopy. Persistent mineralization of weddellite is observed with altering key experimental parameters, including pH, Ca2+ and PA concentrations. In particular, the initial concentration of PA can significantly influence the morphology of weddellite crystal. Oxalate acid is commonly of biological origin; thus calcium oxalate is considered to be a biomarker. However, our results reveal that calcium oxalate can be harvested by a UV photolysis pathway. Moreover, prebiotic sources of organics (e.g., PA, glycine, alanine, and aspartic acid) have been proven to be available through abiotic pathways. Therefore, our results may provide a new abiotic pathway of calcium oxalate formation. Considering that calcium oxalate minerals have been taken as biosignatures for the origin and early evolution of life on Earth and astrobiological investigations, its formation and accumulation by the photolysis of abiological organic compounds should be taken into account.
Asunto(s)
Medio Ambiente Extraterrestre , Marte , Biomarcadores , Oxalato de Calcio/química , Planeta Tierra , ExobiologíaRESUMEN
Mesothelin (MSLN) overexpression in pancreatic cancer (PC) leads to enhanced cell survival/proliferation and tumor progression. After screening for a number of growth factors/cytokines, we found that the MSLN expression correlated closely with interleukin (IL)-6 in human PC specimens and cell lines. Stably overexpressing MSLN in different PC cell lines (MIA-MSLN and Panc1-MSLN) led to higher IL-6 production. Silencing MSLN by small interfering RNA (siRNA) significantly reduced IL-6 levels. Blocking the observed constitutive activation of nuclear factor-kappaB (NF-κB) with IKK inhibitor wedelolactone in MIA-MSLN cells also reduced IL-6. Silencing IL-6 by siRNA reduced cell proliferation, cell cycle progression and induced apoptosis with significant decrease of c-myc/bcl-2. Interestingly, recombinant IL-6-induced proliferation of MIA-MSLN cells but not MIA-V cells. Although messenger RNA/protein levels of IL-6R did not vary, soluble IL-6R (sIL-6R) was significantly elevated in MIA-MSLN and was reduced by treatment with the TACE/ADAM17 inhibitor TAPI-1, indicating intramembrane IL-6R cleavage and IL-6 trans-signaling may be operative in MIA-MSLN cells. Blocking the IL-6/sIL-6R axis using sIL-6R antibody abrogated basal proliferation/survival as well as recombinant human IL-6-induced cell proliferation. Our data suggest that MSLN-activated NF-κB induces elevated IL-6 expression, which acts as a growth factor to support PC cell survival/proliferation through a novel auto/paracrine IL-6/sIL-6R trans-signaling. In addition, using a panel of PC cells with varying MSLN/IL-6 expressions, we showed that MSLN/IL-6 axis is a major survival axis in PC supporting tumor cell growth under anchorage-dependent and independent conditions. The close correlation between MSLN and IL-6 provides a new rationale for combination therapy for effective control of MSLN-overexpressing PCs.