RESUMEN
Stress granules (SGs) are functional messenger ribonucleoprotein aggregates, and their assembly is an important cellular process required for remodeling the signaling network to cope with extensive environmental stresses. SG formation is a stepwise process that involves the formation of a stable core followed by a less stable outer shell, and this process is often hampered by faulty regulation of protein phosphorylation. It remains unclear, however, which kinase activity is essential for SG formation. Here, we screened small molecule library of kinase inhibitors using a well-validated fluorogenic SG probe. Our screen, time-lapse microscopy, and biochemical analyses identified an ATP-mimetic SG inhibitor that selectively interferes with the fusion and growth, rather than the initial assembly, of SG core structures into the large assemblies. Thus, SGs utilize ATP-dependent chemical reactions to achieve their functional architectures.
Asunto(s)
Adenosina Trifosfato/metabolismo , Células Cultivadas , Fluorescencia , Células HEK293 , Células HeLa , Humanos , Inhibidores de Proteínas Quinasas/farmacología , Bibliotecas de Moléculas Pequeñas/farmacología , Estrés Fisiológico/efectos de los fármacosRESUMEN
Rapid endoplasmic reticulum (ER) stress-induced export (RESET) is undoubtedly beneficial in that it eliminates misfolded prion protein (PrP) from a stressed ER. Considering that RESET induces rapid endocytosis of misfolded PrP for degradation, it is questionable whether RESET is beneficial when its product amount overwhelms the capacity of subsequent clearance pathways. We require a strategy to monitor the endocytic flux rate of misfolded PrPs. Here, we stabilized misfolded PrPs by inserting red fluorescent protein (RFP) and indirectly determined this rate by monitoring the lysosomal free RFP. We discovered a surveillance mechanism that limits endocytosis of misfolded PrPs through plasma membrane quality control (pmQC). pmQC was regulated by the thiol-disulfide status of misfolded PrPs and consequently accumulates nonpathogenic PrP variants at the plasma membrane. This variant alleviated prion proteotoxicity induced by persistent RESET. Thus, PrP endocytosis is regulated by pmQC to ensure the safety of endolysosomal pathway from persistent internalization of misfolded PrP.-Lee, D., Lee, S., Shin, Y., Song, Y., Kang, S.-W. Thiol-disulfide status regulates quality control of prion protein at the plasma membrane.
Asunto(s)
Membrana Celular/efectos de los fármacos , Disulfuros/farmacología , Proteínas Priónicas/metabolismo , Compuestos de Sulfhidrilo/farmacología , Línea Celular Tumoral , Membrana Celular/metabolismo , Endocitosis/efectos de los fármacos , Retículo Endoplásmico/efectos de los fármacos , Estrés del Retículo Endoplásmico/efectos de los fármacos , Células HeLa , Humanos , Lisosomas/efectos de los fármacos , Lisosomas/metabolismo , Enfermedades por Prión/metabolismo , Pliegue de Proteína/efectos de los fármacos , Transporte de Proteínas/efectos de los fármacos , Control de CalidadRESUMEN
DISCLAIMER: In an effort to expedite the publication of articles, AJHP is posting manuscripts online as soon as possible after acceptance. Accepted manuscripts have been peer-reviewed and copyedited, but are posted online before technical formatting and author proofing. These manuscripts are not the final version of record and will be replaced with the final article (formatted per AJHP style and proofed by the authors) at a later time. PURPOSE: To evaluate the effect of oncology services rendered by clinical pharmacists on reducing chemotherapy-induced nausea and vomiting (CINV) and improving overall treatment experiences. METHODS: A systematic review and meta-analysis were conducted using studies retrieved from PubMed, Embase, Cochrane Central Register of Controlled Trials (CENTRAL), and Research Information Sharing Service (RISS). The incidence and severity of CINV were evaluated as primary outcomes. Secondary outcomes were patient adherence, patient satisfaction, quality of life (QoL), emergency department (ED) visits, hospitalizations, and costs. RESULTS: A total of 12 studies were selected for systematic review, with 8 studies eligible for meta-analysis. We found that clinical pharmacy services contributed to preventing and alleviating CINV as well as improving patient's medication adherence, treatment satisfaction, and QoL, reducing hospital visits, and achieving cost savings. In the meta-analysis, pharmacists' interventions were notably effective in reducing the incidence of nausea (odds ratio [OR], 1.917; 95% CI, 1.243-2.955; P = 0.003) and vomiting (OR, 2.491; 95% CI, 1.199-5.177; P = 0.014) during overall treatments periods relative to results in control groups. In addition, the impact of clinical pharmacy services on CINV control was greater during the delayed phase compared to the acute phase. CONCLUSION: This study demonstrated the important role of clinical pharmacy services in controlling CINV and enhancing the overall treatment experience for patients with cancer. Further studies with standardized pharmacists' services and outcome measures are needed to validate our findings.
RESUMEN
BACKGROUND/AIM: Neurogenesis is an important process in the recovery from neurological damage caused by ischemic lesions. Endogenous neurogenesis is insufficient to restore neuronal damage following cerebral ischemia. Dexmedetomidine (DEX) exerts neuroprotective effects against cerebral ischemia and ischemia/reperfusion injury. DEX promotes neurogenesis, including neuronal proliferation and maturation in the hippocampus. In a previous study, we showed that early neurogenesis increased 3 days after bilateral common carotid artery occlusion (BCCAO). In this study, we investigated the effect of DEX on neurogenesis 3 days after BCCAO. MATERIALS AND METHODS: Male Sprague-Dawley (SD) rats (7-8 weeks old) were used as a BCCAO model. Right and left common carotid arteries of the rats were occluded using 4-0 silk sutures. Two hours after surgery, an intracranial DEX injection was administered to rats that underwent surgery using a stereotaxic injector. Brains were obtained from control and BCCAO rats 3 days after surgery. Immunohistochemistry was performed on the cortex and dentate gyrus of the hippocampus using a NeuN antibody. Western blot was performed with HIF1α and brain-derived neurotrophic factor (BDNF) antibodies. RESULTS: The number of mature neurons decreased 3 days after BCCAO, but DEX treatment alleviated neural loss in the parietal cortex and hippocampus. Up-regulation of BDNF was also observed after dexmedetomidine treatment. CONCLUSION: Stereotaxic injection of dexmedetomidine alleviates neural loss following BCCAO by up-regulating BDNF expression.
Asunto(s)
Isquemia Encefálica , Dexmedetomidina , Ratas , Masculino , Animales , Factor Neurotrófico Derivado del Encéfalo/genética , Ratas Sprague-Dawley , Regulación hacia Arriba , Dexmedetomidina/farmacología , Dexmedetomidina/metabolismo , Isquemia Encefálica/tratamiento farmacológico , Isquemia Encefálica/etiología , Isquemia Encefálica/metabolismo , Hipocampo/metabolismo , Arteria Carótida Común/metabolismoRESUMEN
Misfolded glycosylphosphatidylinositol-anchored prion protein (PrP) is primarily degraded in lysosomes but is often rapidly removed from the cell surface before endocytosis in a preemptive manner. However, this mechanism is poorly understood. In this study, we discovered a disease-causing prion mutation (Q212P) that exceptionally promoted the extracellular release of PrP. Spatiotemporal analyses combined with genome editing identified the role of sheddase ADAM10 in Q212P shedding from the cell surface. ADAM10 was observed to catalytically interacts with Q212P but non-catalytically with wild-type PrP (wtPrP). This intrinsic difference in the interaction of ADAM10 between Q212P and wtPrP allowed Q212P to selectively access the sheddase activity of ADAM10 in a redox-sensitive manner. In addition, redox perturbation instigated the latent misfolding propensity of Q212P and disrupted the catalytic interaction between PrP and ADAM10, resulting in the accumulation of misfolded PrP on the cell surface. Upon recovery, active ADAM10 was able to reversibly release the surface Q212P. However, it might prove detrimental if unregulated resulting in unexpected proteotoxicity. This study provides a molecular basis of the mutant-selective shedding of PrP by demonstrating the catalytic interaction of ADAM10 with Q212P.
Asunto(s)
Proteínas Priónicas , Priones , Proteína ADAM10/genética , Proteína ADAM10/metabolismo , Glicosilfosfatidilinositoles , Oxidación-Reducción , Proteínas Priónicas/genética , Proteínas Priónicas/metabolismo , Priones/genética , Priones/metabolismoRESUMEN
Protein import across the endoplasmic reticulum membrane is physiologically regulated in a substrate-selective manner to ensure the protection of stressed ER from the overload of misfolded proteins. However, it is poorly understood how different types of substrates are accurately distinguished and disqualified during translocational regulation. In this study, we found poorly assembled translocon-associated protein (TRAP) complexes in stressed ER. Immunoaffinity purification identified calnexin in the TRAP complex in which poor assembly inhibited membrane insertion of the prion protein (PrP) in a transmembrane sequence-selective manner, through translocational regulation. This reaction was induced selectively by redox perturbation, rather than calcium depletion, in the ER. The liberation of ERp57 from calnexin appeared to be the reason for the redox sensitivity. Stress-independent disruption of the TRAP complex prevented a pathogenic transmembrane form of PrP (ctmPrP) from accumulating in the ER. This study uncovered a previously unappreciated role for calnexin in assisting the redox-sensitive function of the TRAP complex and provided insights into the ER stress-induced reassembly of translocon auxiliary components as a key mechanism by which protein translocation acquires substrate selectivity.
Asunto(s)
Proteínas de Unión al Calcio/metabolismo , Estrés del Retículo Endoplásmico/genética , Retículo Endoplásmico/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas Priónicas/química , Proteínas Priónicas/metabolismo , Receptores Citoplasmáticos y Nucleares/metabolismo , Receptores de Péptidos/metabolismo , Animales , Sistemas CRISPR-Cas , Proteínas de Unión al Calcio/genética , Calnexina/metabolismo , Línea Celular , Cricetinae , Edición Génica , Humanos , Glicoproteínas de Membrana/genética , Proteínas Mutantes/química , Proteínas Mutantes/metabolismo , Oxidación-Reducción , Proteínas Priónicas/genética , Transporte de Proteínas , Receptores Citoplasmáticos y Nucleares/genética , Receptores de Péptidos/genética , Especificidad por SustratoRESUMEN
Slot-die coating plays an important role in printed electronics, which are fabricated by stacking thin films and patterns. As electronic devices are being required to have higher performance, the importance of coating uniformity cannot be overestimated in the slot-die coating. The coating uniformity consists of two directions: nozzle direction, which is affected by the interior design of the head, and machine direction, which is majorly related to exterior operating conditions. In this research, the empirical design procedure of a slot-die head is proposed. The internal resistance values of the slot-die were calculated using the experimental parameter obtained through a coating experiment, and the acquired resistances were reflected in the modeling of the interior design of the slot-die head. The hanger type reservoir was adapted to minimize the consumption of ink. After fabricating the slot-die head, coating experiments were carried out using PEDOT:PSS ink. The resulting deviation of the coating thickness was within ±1.7%, thus proving that the proposed design predicted the uniformity of the actual thickness of the coating correctly. The significance of the slot-die design method presented in this paper is that it is based on the experimental equation that can be readily applied to the printed electronics industry.
RESUMEN
In this study, antibody responses after equine influenza vaccination were investigated among 1,098 horses in Korea using the hemagglutination inhibition (HI) assay. The equine influenza viruses, A/equine/South Africa/4/03 (H3N8) and A/equine/Wildeshausen/1/08 (H3N8), were used as antigens in the HI assay. The mean seropositive rates were 91.7% (geometric mean antibody levels (GMT), 56.8) and 93.6% (GMT, 105.2) for A/equine/South Africa/4/03 and A/equine/Wildeshausen/1/08, respectively. Yearlings and two-year-olds in training exhibited lower positive rates (68.1% (GMT, 14) and 61.7% (GMT, 11.9), respectively, with different antigens) than average. Horses two years old or younger may require more attention in vaccination against equine influenza according to the vaccination regime, because they could be a target of the equine influenza virus.
Asunto(s)
Anticuerpos Antivirales/sangre , Enfermedades de los Caballos/prevención & control , Vacunas contra la Influenza/inmunología , Infecciones por Orthomyxoviridae/veterinaria , Vacunación , Envejecimiento , Animales , Pollos , Eritrocitos , Femenino , Enfermedades de los Caballos/sangre , Caballos , Subtipo H3N8 del Virus de la Influenza A/inmunología , Masculino , Infecciones por Orthomyxoviridae/sangre , Infecciones por Orthomyxoviridae/epidemiología , Infecciones por Orthomyxoviridae/prevención & control , República de Corea/epidemiologíaRESUMEN
Hepatitis C virus (HCV) RNA replication requires cellular factors as well as viral non-structural proteins (NS protein). Using small interfering RNA (siRNA) library screening, we previously identified c-Fos as a host factor involved in HCV propagation. In the present study, we demonstrated that silencing of c-Fos expression resulted in decrease of HCV propagation in cell culture grown HCV (HCVcc)-infected cells; whereas overexpression of c-Fos significantly increased HCV propagation. We further confirmed the positive role of c-Fos in HCV propagation by both HCV-luciferase reporter assay and immunofluorescence analysis. We showed that c-Fos level was upregulated by HCV infection. Furthermore, phorbol 12-myristate 13-acetate (PMA)-induced c-Fos level was synergistically increased by HCV infection. These data suggest that c-Fos acts as a positive regulator of HCV propagation and may contribute to HCV-associated pathogenesis.