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1.
Clin J Gastroenterol ; 16(6): 871-876, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37581719

RESUMEN

Cat scratch disease (CSD) is associated with Bartonella henselae (B. henselae) infection caused by cat scratches or bites. It typically presents with lymphadenitis and fever. However, there are atypical cases such as hepatosplenic CSD, which presents with specific lesions in the liver and spleen. Hemophagocytic lymphohistiocytosis (HLH) is a rare and severe multisystem disorder triggered by infections, cancers, or autoimmune diseases. We experienced a rare case of hepatosplenic CSD with HLH in a non-immunocompromised adult. A 78-year-old woman complained of fever and fatigue. Laboratory tests revealed anemia and liver dysfunction; abdominal contrast-enhanced computed tomography (CT) revealed splenomegaly and nodular hypodense areas in the spleen. In addition, the levels of ferritin and serum soluble IL-2R were markedly elevated, so clinical diagnosis of HLH was made. Positron emission tomography/CT revealed diffuse fluorodeoxyglucose uptake in the liver and spleen suggesting malignant lymphoma, while the pathological findings from liver biopsy suggested infectious diseases. Although she had no cat bites and scratches, she had many cats; therefore, serum B. henselae antibody titers were measured. The B. henselae IgG and IgM titer were 1:128 and 1:20; thus, she was diagnosed with hepatosplenic CSD. Patients with hepatosplenic nodular lesions and contact with cats should be considered for this disease.


Asunto(s)
Bartonella henselae , Enfermedad por Rasguño de Gato , Hepatopatías , Linfohistiocitosis Hemofagocítica , Adulto , Femenino , Humanos , Gatos , Animales , Anciano , Enfermedad por Rasguño de Gato/complicaciones , Enfermedad por Rasguño de Gato/diagnóstico , Enfermedad por Rasguño de Gato/patología , Linfohistiocitosis Hemofagocítica/complicaciones , Linfohistiocitosis Hemofagocítica/diagnóstico , Hepatopatías/complicaciones , Hepatopatías/diagnóstico por imagen , Anticuerpos Antibacterianos
2.
Medicine (Baltimore) ; 102(25): e34118, 2023 Jun 23.
Artículo en Inglés | MEDLINE | ID: mdl-37352037

RESUMEN

RATIONALE: Tocilizumab, a humanized anti-interleukin-6 (IL-6) receptor monoclonal antibody, is used for the treatment of adult-onset Still disease (AOSD). Despite its efficacy in many clinical situations, concerns have been raised regarding intestinal mucosal injury in patients receiving tocilizumab. PATIENT CONCERNS: A 64-year-old woman with a history of AOSD was admitted to our hospital with hematochezia. She had AOSD for 15 years and underwent treatment with biweekly tocilizumab 9 months prior to admission. Colonoscopy revealed a large punched-out ulcer in the terminal ileum. On pathological evaluation, nonspecific enteritis with lymphocytes and eosinophils were seen. Based on the location and shape of the lesion, we suspected intestinal Behçet's disease. However, the ulcer reduced in size over time by discontinuation of tocilizumab without additional drug treatment, indicating that it was a drug-induced ulcer. DIAGNOSIS: The patient was diagnosed with tocilizumab-induced small intestinal ulcer. INTERVENTIONS: The patient treated with the discontinuation of tocilizumab. OUTCOMES: The discontinuation of tocilizumab resulted in ulcer scarring. There was no recurrence of hematochezia. LESSONS: Tocilizumab can cause deep ulcerative lesions in the terminal ileum, which may resemble intestinal Behçet's disease. It is important to continuously monitor abdominal symptoms during tocilizumab therapy and aggressively perform colonoscopy when hematochezia or abdominal pain is observed.


Asunto(s)
Síndrome de Behçet , Enfermedades Intestinales , Adulto , Femenino , Humanos , Persona de Mediana Edad , Síndrome de Behçet/tratamiento farmacológico , Úlcera/inducido químicamente , Úlcera/diagnóstico , Úlcera/tratamiento farmacológico , Enfermedades Intestinales/inducido químicamente , Enfermedades Intestinales/diagnóstico , Enfermedades Intestinales/tratamiento farmacológico , Anticuerpos Monoclonales/uso terapéutico , Íleon/patología , Hemorragia Gastrointestinal/tratamiento farmacológico
3.
Sci Signal ; 16(773): eabn0782, 2023 02 21.
Artículo en Inglés | MEDLINE | ID: mdl-36809024

RESUMEN

Insulin regulates various cellular metabolic processes by activating specific isoforms of the Akt family of kinases. Here, we elucidated metabolic pathways that are regulated in an Akt2-dependent manner. We constructed a transomics network by quantifying phosphorylated Akt substrates, metabolites, and transcripts in C2C12 skeletal muscle cells with acute, optogenetically induced activation of Akt2. We found that Akt2-specific activation predominantly affected Akt substrate phosphorylation and metabolite regulation rather than transcript regulation. The transomics network revealed that Akt2 regulated the lower glycolysis pathway and nucleotide metabolism and cooperated with Akt2-independent signaling to promote the rate-limiting steps in these processes, such as the first step of glycolysis, glucose uptake, and the activation of the pyrimidine metabolic enzyme CAD. Together, our findings reveal the mechanism of Akt2-dependent metabolic pathway regulation, paving the way for Akt2-targeting therapeutics in diabetes and metabolic disorders.


Asunto(s)
Optogenética , Proteínas Proto-Oncogénicas c-akt , Proteínas Proto-Oncogénicas c-akt/metabolismo , Músculo Esquelético/metabolismo , Transducción de Señal , Fosforilación , Insulina/metabolismo , Redes y Vías Metabólicas
4.
Intern Med ; 62(10): 1501-1506, 2023 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-36171126

RESUMEN

Eosinophilic granulomatosis with polyangiitis (EGPA) is a systemic disorder characterized by tissue eosinophilic infiltration and vasculitis. Although EGPA causes multiple organ damage, it causes cholecystitis less frequently. We herein report a case of acute cholecystitis associated with EGPA in which successful treatment with glucocorticoid therapy allowed surgery to be avoided. EGPA can present as acute cholecystitis. It is important not to overlook acute cholecystitis associated with EGPA in patients with abdominal pain with peripheral eosinophilia. Furthermore, in cases of mild cholecystitis associated with EGPA that are diagnosed preoperatively, cholecystectomy might be avoided with conservative glucocorticoid treatment.


Asunto(s)
Colecistitis Aguda , Colecistitis , Síndrome de Churg-Strauss , Eosinofilia , Granulomatosis con Poliangitis , Humanos , Granulomatosis con Poliangitis/complicaciones , Granulomatosis con Poliangitis/tratamiento farmacológico , Granulomatosis con Poliangitis/diagnóstico , Síndrome de Churg-Strauss/diagnóstico , Glucocorticoides/uso terapéutico , Colecistitis Aguda/complicaciones , Colecistitis Aguda/tratamiento farmacológico , Colecistitis/complicaciones , Colecistitis/tratamiento farmacológico , Eosinofilia/complicaciones , Eosinofilia/tratamiento farmacológico
5.
Cells ; 11(19)2022 10 05.
Artículo en Inglés | MEDLINE | ID: mdl-36231098

RESUMEN

The application of NIR to optogenetic systems is in great demand due to its superior properties enabling in vivo deep tissue penetration. Irradiation of NIR to tissue samples or cells rapidly generates heat locally. The resultant elevation in temperature affects cells at the molecular level because of the activation of the heat shock pathway and ROS generation. Nevertheless, few reports have presented detailed comparisons of the effects of the temperature change rate on signaling pathway biomolecules, especially those of rapid heat changes. Aiming at broadening the understanding of temperature sensitivity, we investigated seven insulin signaling pathway biomolecules (INSR, IRS1, Akt, GSK3ß, p70S6K, FoxO1, and ERK1/2) in three cell lines (C2C12, HepG2, and Fao) at temperatures between 25 and 45 °C. The results show that, except for INSR, pAkt(T308), and FoxO1, biomolecules are sensitive to rapid temperature changes at temperatures higher than 42 °C, at which they are significantly phosphorylated. At 25 °C, around a 50% reduction in phosphorylation occurred. Moreover, p70S6K is sensitive over time. It dephosphorylates quickly (5 min) and then phosphorylates over time. Our findings extend the temperature range to 45 °C, while providing additional time course information about the signaling pathway biomolecule response necessary to advance NIR optogenetic research.


Asunto(s)
Insulina , Proteínas Quinasas S6 Ribosómicas 70-kDa , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Insulina/metabolismo , Optogenética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Especies Reactivas de Oxígeno , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismo , Transducción de Señal , Temperatura
6.
Nat Commun ; 13(1): 4288, 2022 08 10.
Artículo en Inglés | MEDLINE | ID: mdl-35948553

RESUMEN

Photoluminescent gold clusters are functionally variable chemical modules by ligand design. Chemical modification of protective ligands and introduction of different metals into the gold clusters lead to discover unique chemical and physical properties based on their significantly perturbed electronic structures. Here we report the synthesis of carbon-centered Au(I)-Ag(I) clusters with high phosphorescence quantum yields using N-heterocyclic carbene ligands. Specifically, a heterometallic cluster [(C)(AuI-L)6AgI2]4+, where L denotes benzimidazolylidene-based carbene ligands featuring N-pyridyl substituents, shows a significantly high phosphorescence quantum yield (Φ = 0.88). Theoretical calculations suggest that the carbene ligands accelerate the radiative decay by affecting the spin-orbit coupling, and the benzimidazolylidene ligands further suppress the non-radiative pathway. Furthermore, these clusters with carbene ligands are taken up into cells, emit phosphorescence and translocate to a particular organelle. Such well-defined, highly phosphorescent C-centered Au(I)-Ag(I) clusters will enable ligand-specific, organelle-selective phosphorescence imaging and dynamic analysis of molecular distribution and translocation pathways in cells.


Asunto(s)
Oro , Metano , Oro/química , Ligandos , Metano/análogos & derivados , Metano/química , Orgánulos
7.
Cancer Diagn Progn ; 2(4): 471-481, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35813008

RESUMEN

AIM: The aim of this study was to clarify the usefulness of the Liver Imaging Reporting and Data System (LI-RADS) for predicting a patient's prognosis after transcatheter arterial chemoembolization (TACE) combined with radiofrequency ablation (TACE-RFA) for hepatocellular carcinoma (HCC) of Barcelona-Clinic Liver Cancer (BCLC) stage 0 or A. PATIENTS AND METHODS: We retrospectively analyzed cases of patients with HCC who underwent TACE-RFA (Jan 2005 to Dec 2015). Nodules were categorized based on their LI-RADS v2018 core. The LI-RADS category was assigned to each nodule using dynamic contrast-enhanced computed tomography. LR-3, LR-4 and LR-5 nodules were extracted. The overall (OS) and recurrence-free (RFS) survival was assessed among patients with BCLC 0 and BCLC A disease. RESULTS: Of the 64 nodules extracted, 22 were LR-3 or -4 (mean±standard deviation=14.8±6.7 mm) and 42 were LR-5 (17.1±6.9 mm). Regarding OS, there was no significant difference between those with LR-3 or -4 and LR-5 (p=0.278). In terms of RFS, there was a significant difference between those with LR-3 or -4 and those with LR-5 (p=0.03). In particular, patients with BCLC A with LR-5 nodules had significantly poorer RFS than those with LR-3 or -4 (p=0.016) nodules. CONCLUSION: For patients with BCLC A, LR-3 or -4 nodules are associated with a better prognosis than LR-5 nodules.

8.
Cells ; 12(1)2022 12 20.
Artículo en Inglés | MEDLINE | ID: mdl-36611804

RESUMEN

The plasma membrane (PM) serves multiple functions to support cell activities with its heterogeneous molecular distribution. Fatty acids (FAs) are hydrophobic components of the PM whose saturation and length determine the membrane's physical properties. The FA distribution contributes to the PM's lateral heterogeneity. However, the distribution of PM FAs is poorly understood. Here, we proposed the FA cluster hypothesis, which suggested that FAs on the PM exist as clusters. By the optogenetic tool translocating the endoplasmic reticulum (ER), we were able to manipulate the distribution of PM FAs. We used time-of-flight combined secondary ion mass spectrometry (TOF-SIMS) to image PM FAs and discovered that PM FAs were presented and distributed as clusters and are also manipulated as clusters. We also found the existence of multi-FA clusters formed by the colocalization of more than one FA. Our optogenetic tool also decreased the clustering degree of FA clusters and the formation probability of multi-FA clusters. This research opens up new avenues and perspectives to study PM heterogeneity from an FA perspective. This research also suggests a possible treatment for diseases caused by PM lipid aggregation and furnished a convenient tool for therapeutic development.


Asunto(s)
Ácidos Grasos , Espectrometría de Masa de Ion Secundario , Ácidos Grasos/metabolismo , Espectrometría de Masa de Ion Secundario/métodos , Optogenética , Membrana Celular/metabolismo , Diagnóstico por Imagen
9.
World J Oncol ; 12(5): 165-172, 2021 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-34804279

RESUMEN

BACKGROUND: Although lenvatinib treatment has a favorable efficacy for unresectable hepatocellular carcinoma (HCC), it is associated with adverse events (AEs) that must be closely monitored and managed. Thrombocytopenia is one of the major AEs. The aim of this study was to clarify whether thrombocytopenia can be predicted by the plasma concentration of lenvatinib. METHODS: This was a single-center retrospective observational study. Twenty-three patients with unresectable HCC and pharmacokinetics data at the initial lenvatinib administration between May 2018 and September 2020 at Oita University Hospital were enrolled. The AEs during the 4 weeks after the initiation of treatment were evaluated, and the correlations between the thrombocytopenia and the plasma concentration of lenvatinib were examined. Spearman's correlation was used to evaluate the correlation between two continuous variables. RESULTS: The rate of platelet count decrease correlated with the maximum plasma concentration (Cmax) (r = 0.65, P = 0.001), whereas it did not with the minimum plasma concentration (Cmin) (r = 0.29, P = 0.206). After stepwise multiple linear regression analysis, the starting dose of lenvatinib and the serum albumin concentration were identified as independent explanatory variables. Next, a formula for predicting the Cmax using these two variables was created. The predicted Cmax was strongly correlated with the Cmax (r = 0.87, P < 0.0001) and the rate of platelet count decrease (r = 0.67, P = 0.001). CONCLUSIONS: This study identified the usefulness of the drug Cmax to predict the rate of platelet count decrease within 4 weeks after the initiation of treatment. Although it is difficult to measure the plasma concentration of lenvatinib in community hospitals, the predicted Cmax is useful for predicting the rate of platelet count decrease with this treatment.

10.
Ther Drug Monit ; 43(5): 664-671, 2021 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-34521802

RESUMEN

BACKGROUND: Lenvatinib is increasingly being selected as the first-line treatment for unresectable hepatocellular carcinoma (HCC) based on the results of the REFLECT trial. However, early discontinuation of lenvatinib because of adverse effects is a frequent occurrence. Hence, lenvatinib is a difficult drug for use in the clinical setting. One of the causes is that the dose of lenvatinib is mainly determined by body weight alone, despite high interindividual variability. To overcome this problem, a dosing regimen of lenvatinib based on a population pharmacokinetic (PPK) model for HCC patients is proposed. The aim of this study was to develop a high-throughput quantification method for lenvatinib using ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) that can be applied to a PPK analysis of HCC patients in the future. METHODS: After a simple solid-phase extraction step using a 96-well plate, lenvatinib was analyzed by UHPLC-MS/MS in a positive electrospray ionization mode. RESULTS: The novel method fulfilled the requirements of the US Food and Drug Administration guidance on bioanalytical method validation. The calibration curve was linear over the 0.2-1000 ng/mL concentration range. The average recovery rate was 98.63 ± 4.55% (mean ± SD). The precision was below 6.05%, and the accuracy was within 12.96% for all quality control levels. The matrix effect varied between 103.33% and 134.61%. This assay was successfully applied to the measurement of plasma concentrations in 6 HCC patients receiving lenvatinib. CONCLUSIONS: A novel high-throughput UHPLC-MS/MS assay for quantification of lenvatinib in human plasma was successfully developed. This method can be applied to PPK analysis for patients receiving lenvatinib in the clinical setting.


Asunto(s)
Carcinoma Hepatocelular , Neoplasias Hepáticas , Compuestos de Fenilurea/farmacocinética , Quinolinas/farmacocinética , Carcinoma Hepatocelular/tratamiento farmacológico , Cromatografía Líquida de Alta Presión , Ensayos Analíticos de Alto Rendimiento , Humanos , Neoplasias Hepáticas/tratamiento farmacológico , Compuestos de Fenilurea/administración & dosificación , Quinolinas/administración & dosificación , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem , Estados Unidos
11.
Obes Sci Pract ; 7(1): 91-99, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33680496

RESUMEN

AIM: Antioxidant therapy for with vitamin E appears to be effective for the treatment of nonalcoholic fatty liver disease (NAFLD). However, the mechanism of action and optimal therapeutic dosage is unclear. The present study was undertaken to examine whether the effects of α-tocopherol (α-Toc) on NAFLD are dose-dependent in a diet-induced obese model. METHODS: Male mice were fed standard chow, high-fat (HF) diet, HF diet with low-dose, or with high dose of α-Toc supplementation. Histological findings, triglyceride content, and the levels of protein expression related to fatty acid synthesis/oxidation such as carnitine palmitoyltransferase I (CPT-1) of liver were evaluated. In addition, 2-tetradecylglycidic acid (TDGA), a CPT-1 inhibitor, was administered to mice fed HF diet with low-dose of α-Toc. Finally, HepG2 cells in fat-loaded environment were treated with 0-50 µM α-Toc. RESULTS: Treatment of low-dose of α-Toc decreased HF-induced hepatic fat accumulation, but this finding was not observed in treatment of high dose of α-Toc. HF-induced reduction of CPT-1 was attenuated with low-dose of α-Toc but not with high dose of α-Toc. TDGA suppressed the improvement of histological findings in liver induced by low-dose of α-Toc treatment. CPT-1 expression in HepG2 cells increased in response to low-dose of α-Toc, but not in high dose. CONCLUSIONS: Dual action of α-Toc on CPT-1 protein levels was observed. The effect of vitamin E on NAFLD may be not be dose-dependent.

12.
Adv Exp Med Biol ; 1293: 247-263, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33398818

RESUMEN

In multicellular organisms, living cells cooperate with each other to exert coordinated complex functions by responding to extracellular chemical or physical stimuli via proteins on the plasma membrane. Conventionally, chemical signal transduction or mechano-transduction has been investigated by chemical, genetic, or physical perturbation; however, these methods cannot manipulate biomolecular reactions at high spatiotemporal resolution. In contrast, recent advances in optogenetic perturbation approaches have succeeded in controlling signal transduction with external light. The methods have enabled spatiotemporal perturbation of the signaling, providing functional roles of the specific proteins. In this chapter, we summarize recent advances in the optogenetic tools that modulate the function of a receptor protein. While most optogenetic systems have been devised for controlling ion channel conductivities, the present review focuses on the other membrane proteins involved in chemical transduction or mechano-transduction. We describe the properties of natural or artificial photoreceptor proteins used in optogenetic systems. Then, we discuss the strategies for controlling the receptor protein functions by external light. Future prospects of optogenetic tool development are discussed.


Asunto(s)
Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/efectos de la radiación , Optogenética/métodos , Receptores de Superficie Celular/metabolismo , Receptores de Superficie Celular/efectos de la radiación , Transducción de Señal/efectos de la radiación
13.
Sci Rep ; 10(1): 18768, 2020 10 30.
Artículo en Inglés | MEDLINE | ID: mdl-33127939

RESUMEN

Non-alcoholic fatty liver disease (NAFLD) is a significant problem because its prevalence is increasing worldwide. Recent animal studies have identified gut microbiota as a potentially important player in the pathogenesis of NAFLD. Previously, we reported that the administration of branched-chain amino acids (BCAAs) reduces hepatic fat accumulation in experimental animal models. This study aimed to clarify how changes in the intestinal microbial flora following the administration of BCAAs affect a high-fat diet (HF)-induced fat accumulation in the liver. We examined whether the administration of BCAAs alters the development of hepatic fat accumulation as well as intestinal microbial flora. The oral administration of BCAAs (3% kcal) induced a significant increase in Ruminococcus flavefaciens (R. flavefaciens) and portal acetic acid levels, and it reduced hepatic fat accumulation in HF-fed rats. In addition, BCAAs reduced the expression of the lipogenesis-related genes FAS and ACC in the liver. Furthermore, we observed that R. flavefaciens is essential for promoting a BCAA-induced reduction in hepatic fat accumulation. These data suggest that BCAA treatment induces the proliferation of intestinal flora including R. flavefaciens and that portal acetic acid synthesized from intestinal flora improves NAFLD by downregulating the expression of FAS and ACC in the liver.


Asunto(s)
Ácido Acético/metabolismo , Aminoácidos de Cadena Ramificada/administración & dosificación , Suplementos Dietéticos , Grasas/metabolismo , Microbioma Gastrointestinal , Hígado/metabolismo , Animales , Dieta Alta en Grasa , Lipogénesis , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Enfermedad del Hígado Graso no Alcohólico/patología , Ratas
14.
Int J Mol Sci ; 21(18)2020 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-32906768

RESUMEN

In vivo bioluminescence imaging (BLI), which is based on luminescence emitted by the luciferase-luciferin reaction, has enabled continuous monitoring of various biochemical processes in living animals. Bright luminescence with a high signal-to-background ratio, ideally red or near-infrared light as the emission maximum, is necessary for in vivo animal experiments. Various attempts have been undertaken to achieve this goal, including genetic engineering of luciferase, chemical modulation of luciferin, and utilization of bioluminescence resonance energy transfer (BRET). In this review, we overview a recent advance in the development of a bioluminescence system for in vivo BLI. We also specifically examine the improvement in bioluminescence intensity by mutagenic or chemical modulation on several beetle and marine luciferase bioluminescence systems. We further describe that intramolecular BRET enhances luminescence emission, with recent attempts for the development of red-shifted bioluminescence system, showing great potency in in vivo BLI. Perspectives for future improvement of bioluminescence systems are discussed.


Asunto(s)
Diagnóstico por Imagen/métodos , Sustancias Luminiscentes/química , Mediciones Luminiscentes/métodos , Animales , Pruebas Diagnósticas de Rutina/métodos , Transferencia Resonante de Energía de Fluorescencia/métodos , Humanos , Rayos Infrarrojos , Luciferasas/química , Luciferasas/metabolismo , Luminiscencia , Sustancias Luminiscentes/metabolismo , Espectroscopía Infrarroja Corta/métodos
15.
ACS Chem Biol ; 14(10): 2206-2214, 2019 10 18.
Artículo en Inglés | MEDLINE | ID: mdl-31503442

RESUMEN

Precise integration of individual cell behaviors is indispensable for collective tissue morphogenesis and maintenance of tissue integrity. Organized multicellular behavior is achieved via mechanical coupling of individual cellular contractility, mediated by cell adhesion molecules at the cell-cell interface. Conventionally, gene depletion or laser microsurgery has been used for functional analysis of intercellular mechanotransduction. Nevertheless, these methods are insufficient to investigate either the spatiotemporal dynamics or the biomolecular contribution in cell-cell mechanical coupling within collective multicellular behaviors. Herein, we present our effort in adaption of PhoCl for attenuation of cell-to-cell tension transmission mediated by E-cadherin. To release intercellular contractile tension applied on E-cadherin molecules with external light, a genetically encoded photocleavable module called PhoCl was inserted into the intracellular domain of E-cadherin, thereby creating photocleavable cadherin (PC-cadherin). In response to light illumination, the PC-cadherin cleaved into two fragments inside cells, resulting in attenuating mechanotransduction at intercellular junctions in living epithelial cells. Light-induced perturbation of the intercellular tension balance with surrounding cells changed the cell shape in an epithelial cell sheet. The method is expected to enable optical manipulation of force-mediated cell-to-cell communications in various multicellular behaviors, which contributes to a deeper understanding of embryogenesis and oncogenesis.


Asunto(s)
Antígenos CD/metabolismo , Cadherinas/metabolismo , Mecanotransducción Celular/efectos de la radiación , Proteínas Recombinantes de Fusión/metabolismo , Actomiosina/metabolismo , Animales , Anticuerpos Monoclonales/inmunología , Antígenos CD/inmunología , Antígenos CD/efectos de la radiación , Cadherinas/inmunología , Cadherinas/efectos de la radiación , Comunicación Celular , Perros , Células Epiteliales/citología , Fluorescencia , Humanos , Luz , Proteínas Luminiscentes/metabolismo , Proteínas Luminiscentes/efectos de la radiación , Células MCF-7 , Células de Riñón Canino Madin Darby , Microscopía Confocal , Microscopía Fluorescente , Proteínas Recombinantes de Fusión/efectos de la radiación , Proteína Fluorescente Roja
16.
Anal Sci ; 35(8): 835-838, 2019 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-31281129

RESUMEN

Glucose transporter 4 (GLUT4) is an insulin-regulated glucose transporter, which is vital for blood glucose homeostasis. To clarify the physiological roles of GLUT4, quantitative measurement of GLUT4 exocytosis is indispensable. Herein, we show a rapid detection system for GLUT4 on the cell surface using spontaneous split-luciferase reconstitution. Upon insulin-induced GLUT4 exocytosis, GLUT4 was exposed outside, where luciferase is reconstituted and emitted luminescence. Pretreatment with inhibitors reduced the insulin-induced signal elevation. The results indicate that the developed method is applicable to high-throughput analysis on GLUT4 trafficking, which will greatly accelerate comprehensive research on the physiological roles of GLUT4.


Asunto(s)
Exocitosis , Prueba de Complementación Genética , Transportador de Glucosa de Tipo 4/análisis , Luciferasas/metabolismo , Membrana Celular/metabolismo , Transportador de Glucosa de Tipo 4/metabolismo , Células HEK293 , Ensayos Analíticos de Alto Rendimiento , Humanos , Luciferasas/genética
17.
Nat Commun ; 10(1): 2407, 2019 06 03.
Artículo en Inglés | MEDLINE | ID: mdl-31160592

RESUMEN

The senses play critical roles in helping animals evaluate foods, including fruits that can change both in colour and scent during ripening to attract frugivores. Although numerous studies have assessed the impact of colour on fruit selection, comparatively little is known about fruit scent and how olfactory and visual data are integrated during foraging. We combine 25 months of behavioural data on 75 wild, white-faced capuchins (Cebus imitator) with measurements of fruit colours and scents from 18 dietary plant species. We show that frequency of fruit-directed olfactory behaviour is positively correlated with increases in the volume of fruit odours produced during ripening. Monkeys with red-green colour blindness sniffed fruits more often, indicating that increased reliance on olfaction is a behavioural strategy that mitigates decreased capacity to detect red-green colour contrast. These results demonstrate a complex interaction among fruit traits, sensory capacities and foraging strategies, which help explain variation in primate behaviour.


Asunto(s)
Conducta Animal/fisiología , Defectos de la Visión Cromática/fisiopatología , Visión de Colores/fisiología , Preferencias Alimentarias/fisiología , Frutas , Olfato/fisiología , Animales , Cebus , Odorantes
18.
Hepatol Res ; 48(4): 255-263, 2018 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-29080280

RESUMEN

AIMS: PCR-Invader is a highly sensitive assay for detecting non-structural protein 5A (NS5A) resistance-associated variants (RAVs) of hepatitis C virus (HCV). Here, we validated the accuracy of the semiquantitative PCR-Invader (SQ-PI) assay compared to direct sequencing (DS) for identifying NS5A RAVs, and we evaluated the treatment efficacy of daclatasvir plus asunaprevir (DCV + ASV) for patients judged to be non-positive for NS5A RAVs by SQ-PI. METHODS: Detection rates of NS5A RAVs by SQ-PI and DS were compared for 204 patients with HCV genotype 1b. Patients with non-positive results for NS5A RAVs by SQ-PI were treated by DCV + ASV, and the efficacy of this treatment was examined. RESULTS: All samples judged as negative for NS5A RAVs by SQ-PI were similarly judged by DS. However, 29.7% of samples judged as negative for Y93H by DS were judged as weakly positive or positive by SQ-PI. Among 108 patients who were judged as negative by SQ-PI and treated by DCV + ASV, rates of sustained virologic response at 24 weeks (SVR24) were 96.3% in intention-to-treat analysis and 99.0% in patients who completed treatment. Among patients who were weakly positive for Y93H on SQ-PI, the SVR24 rate was 95.0% (19/20). This rate was 100% (78/78) in patients who were negative for Y93H on SQ-PI and completed treatment. CONCLUSION: Treatment efficacy of DCV + ASV was extremely high among patients who were non-positive for NS5A RAVs on SQ-PI, especially for patients with negative results.

19.
Sci Rep ; 7(1): 17443, 2017 12 12.
Artículo en Inglés | MEDLINE | ID: mdl-29234079

RESUMEN

Post-translational modification by the Small Ubiquitin-related Modifier (SUMO) is indispensable for diverse biological mechanisms. Although various attempts have been made to discover novel SUMO substrate proteins to unveil the roles of SUMOylation, the reversibility of SUMOylation, and the differences in the SUMOylation level still makes it difficult to explore infrequently-SUMOylated proteins in mammalian cells. Here, we developed a method to screen for mammalian SUMOylated proteins using the reconstitution of split fluorescent protein fragments in living mammalian cells. Briefly, the cells harboring cDNAs of SUMOylated proteins were identified by the reconstituted fluorescence emission and separated by cell sorting. The method successfully identified 36 unreported SUMO2-substrate candidates with distinct intracellular localizations and functions. Of the candidates, we found Atac2, a histone acetyltransferase, was SUMOylated at a lysine 408, and further modified by multiple SUMOs without isoform specificity. Because the present method is applicable to other SUMO isoforms and mammalian cell-types, it could contribute to a deeper understanding of the role of SUMOylation in various biological contexts.


Asunto(s)
Técnicas Genéticas , Proteínas/metabolismo , Sumoilación , Células 3T3 , Animales , Western Blotting , Encéfalo/metabolismo , Células HEK293 , Células HeLa , Humanos , Inmunoprecipitación , Ratones , Microscopía Confocal/métodos , Microscopía Fluorescente/métodos , Proteínas/química , Proteínas/genética , Sumoilación/fisiología
20.
J Am Chem Soc ; 138(43): 14347-14353, 2016 11 02.
Artículo en Inglés | MEDLINE | ID: mdl-27726387

RESUMEN

The significant contribution of conventional living polymerization to polymer science assures that living supramolecular polymerization will also lead to a variety of novel phenomena and applications. However, the monomer scope still remains limited in terms of the self-assembly energy landscape; a kinetic trap that retards spontaneous nucleation has to be coupled with a supramolecular polymerization pathway, which is challenging to achieve by molecular design. Herein, we report a rational approach to addressing this issue. We combined the supramolecular polymerization and photoisomerization processes to build the energy landscape, wherein the monomer can be activated/deactivated by light irradiation. In this way, the supramolecular polymerization and kinetic trap can be independently designed in the energy landscape. When the "dormant" monomer was activated by light in the presence of the seed of the supramolecular polymer, the "activated" free monomer was polymerized at the termini of the seed in a chain-growth manner. As a result, we achieved supramolecular polymers with controlled lengths and a narrow polydispersity. Although photoisomerization has been extensively employed in supramolecular polymer chemistry, most studies have focused on the stimuli responsiveness. In this respect, the present study would provoke supramolecular chemists to revisit stimuli-responsive supramolecular polymer systems as potential candidates for devising living supramolecular polymerization.

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