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Cocamidopropyl betaine (CAPB) is a surfactant derived from coconut oil that is widely used in cosmetics and personal products for several purposes, such as a surfactant, foam booster, mildness, and viscosity control. Cocamidopropyl betaine is used at concentrations up to 30% in cosmetics. The acute toxicity, skin irritation, eye irritation, skin sensitization, repeated dose toxicity, genotoxicity, carcinogenicity, and phototoxicity of cocamidopropyl betaine were evaluated. Cocamidopropyl betaine was observed to induce mild skin irritation, eye irritation and skin sensitization. The NOAEL of cocamidopropyl betaine was determined to be 250 mg/kg/day based on the results of a 92-day repeated-dose oral toxicity study in rats. The systemic exposure dose of cocamidopropyl betaine was estimated to range from 0.00120 to 0.93195 mg/kg/day when used in cosmetic products. The margin of safety of cocamidopropyl betaine was calculated to be greater than 100 when used at a maximum concentration of 6% in leave-on products and 30% in rinse-off products, suggesting that its use in cosmetic products is safe under current usage conditions.
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Taste sensors using photonics, termed artificial photonic tongues, have emerged as a promising platform for intuitive taste discrimination. However, the need for complex binding protocols for each taste profile limits their applicability to a narrow range of taste molecules. Here, we introduce an intriguing "binding-free" approach to molecular taste sensing using plasmonics, eliminating the requirement for physical or chemical binding protocols. We develop a wafer-scale plasmonic metasurface constructed by coating metallic nanoparticles in a scalable manner onto a metallic mirror. This metasurface functions to detect molecular refractive indices and surface tensions via 2D projection optical images of an array of liquid droplets containing the taste molecules on top, which can immediately visualize and distinguish between the five basic tastes of molecules (including their mixtures) as well as other additional spicy and alcoholic tastes. We anticipate that this intuitive and rapid taste-sensing approach has the potential to establish a user-friendly and portable taste-sensing platform.
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Adaptive multicolor filters have emerged as key components for ensuring color accuracy and resolution in outdoor visual devices. However, the current state of this technology is still in its infancy and largely reliant on liquid crystal devices that require high voltage and bulky structural designs. Here, we present a multicolor nanofilter consisting of multilayered 'active' plasmonic nanocomposites, wherein metallic nanoparticles are embedded within a conductive polymer nanofilm. These nanocomposites are fabricated with a total thickness below 100 nm using a 'lithography-free' method at the wafer level, and they inherently exhibit three prominent optical modes, accompanying scattering phenomena that produce distinct dichroic reflection and transmission colors. Here, a pivotal achievement is that all these colors are electrically manipulated with an applied external voltage of less than 1 V with 3.5 s of switching speed, encompassing the entire visible spectrum. Furthermore, this electrically programmable multicolor function enables the effective and dynamic modulation of the color temperature of white light across the warm-to-cool spectrum (3250 K-6250 K). This transformative capability is exceptionally valuable for enhancing the performance of outdoor optical devices that are independent of factors such as the sun's elevation and prevailing weather conditions.
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Underwater adhesion processes in nature promise controllable assembly of functional nanoparticles for industrial mass production; However, their artificial strategies have faced challenges to uniformly transfer nanoparticles into a monolayer, particularly those below 100 nm in size, over large areas. Here a scalable "one-shot" self-limiting nanoparticle transfer technique is presented, enabling the efficient transport of nanoparticles from water in microscopic volumes to an entire 2-inch wafer in a remarkably short time of 10 seconds to reach near-maximal surface coverage (≈40%) in a 2D mono-layered fashion. Employing proton engineering in electrostatic assembly accelerates the diffusion of nanoparticles (over 50 µm2/s), resulting in a hundredfold faster coating speed than the previously reported results in the literature. This charge-sensitive process further enables "pick-and-place" nanoparticle patterning at the wafer scale, with large flexibility in surface materials, including flexible metal oxides and 3D-printed polymers. As a result, the fabrication of wafer-scale disordered plasmonic metasurfaces in seconds is successfully demonstrated. These metasurfaces exhibit consistent resonating colors across diverse material and geometrical platforms, showcasing their potential for applications in full-color painting and optical encryption devices.
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PURPOSE: In this study, we aimed to determine the clinicopathologic, radiologic, and molecular significance of the tumor invasiveness to further stratify the patients with high-grade (HG) upper tract urothelial carcinoma (UTUC) who can be treated less aggressively. MATERIALS AND METHODS: Clinicopathologic and radiologic characteristics of 166 surgically resected HG UTUC (48 noninvasive, and 118 invasive) cases were evaluated. Six noninvasive UTUC cases with intratumoral tumor grade heterogeneity were selected for whole-exome sequencing (WES) to understand the underlying molecular pathophysiology. Barcode-tagging sequencing was done for validation of the target genes from WES data. RESULTS: Patients with noninvasive UTUC showed no cancer-specific death with better cancer-specific survival (p < 0.001) and recurrence-free survival (p < 0.001) compared to the patients with invasive UTUC. Compared to the invasive UTUC, noninvasive UTUC was correlated to a low grade (LG) on the preoperative abdominal computed tomography (CT) grading system (p < 0.001), histologic intratumoral tumor grade heterogeneity (p=0.018), discrepancy in preoperative urine cytology diagnosis (p=0.018), and absence of urothelial carcinoma in situ (p < 0.001). WES of the heterogeneous components showed mutually shared HRAS and FGFR3 mutations shared between the HG and LG components. HRAS mutation was associated with the lower grade on preoperative abdominal CT and intratumoral tumor grade heterogeneity (p=0.045 and p < 0.001, respectively), whereas FGFR3 mutation was correlated to the absence of carcinoma in situ (p < 0.001). CONCLUSION: According to our comprehensive analysis, HG noninvasive UTUC can be preoperatively suspected based on distinct preoperative radiologic, cytologic, histologic, and molecular features. Noninvasive HG UTUC shows excellent prognosis and thus should be treated less aggressively.
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Invasividad Neoplásica , Humanos , Masculino , Femenino , Anciano , Persona de Mediana Edad , Secuenciación del Exoma , Carcinoma de Células Transicionales/genética , Carcinoma de Células Transicionales/patología , Carcinoma de Células Transicionales/diagnóstico , Mutación , Pronóstico , Anciano de 80 o más Años , Clasificación del Tumor , Neoplasias Urológicas/diagnóstico , Neoplasias Urológicas/genética , Neoplasias Urológicas/patología , Biomarcadores de Tumor/genética , Receptor Tipo 3 de Factor de Crecimiento de Fibroblastos/genética , Relevancia ClínicaRESUMEN
The misuse of poisonous mushrooms containing amatoxins causes acute liver failure (ALF) in patients and is a cause of significant mortality. Although the toxic mechanisms of α-amanitin (α-AMA) and its interactions with RNA polymerase II (RNAP II) have been studied, α-AMA effector proteins that can interact with α-AMA in hepatocytes have not been systematically studied. Limited proteolysis-coupled mass spectrometry (LiP-MS) is an advanced technology that can quickly identify protein-ligand interactions based on global comparative proteomics. This study identified the α-AMA effector proteins found in human hepatocytes, following the detection of conformotypic peptides using LiP-MS coupled with tandem mass tag (TMT) technology. Proteins that are classified into protein processing in the endoplasmic reticulum and the ribosome during the KEGG pathway can be identified through affinity evaluation, according to α-AMA concentration-dependent LiP-MS and LiP-MS in hepatocytes derived from humans and mice, respectively. The possibility of interaction between α-AMA and proteins containing conformotypic peptides was evaluated through molecular docking studies. The results of this study suggest a novel path for α-AMA to induce hepatotoxicity through interactions with various proteins involved in protein synthesis, as well as with RNAP II.
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Alfa-Amanitina , Hepatocitos , Humanos , Ratones , Animales , Alfa-Amanitina/metabolismo , Alfa-Amanitina/toxicidad , Proteolisis , Simulación del Acoplamiento Molecular , Hepatocitos/metabolismo , Espectrometría de MasasRESUMEN
Mucoepidermoid carcinomas (MECs) are the most common salivary gland malignancy and have a diverse histology. Many histologic variants of MEC have now been confirmed with characteristic molecular alterations involving CRTC1::MAML2 or CRTC1::MAML3 translocations. We here report a series of 7 trabecular variants of MEC which showed a predominant trabecular or nested pattern with either focal glandular differentiation or clear cell change and keloid-like fibrosis in the background. In addition, these tumors were either negative or showed only focal positivity for p63. Such features are not characteristic of known disease entities and resulted in an initial misdiagnosis of adenocarcinoma, not otherwise specified, or low-grade to intermediate-grade MEC with uncertainty. The patients' ages in our cohort ranged from 26 to 55 years with a female predominance (5/7). The tumors were located in the parotid gland (n=3), base of tongue (n=2), hard palate (n=1), and parapharyngeal space (n=1), with a median size of 1.5 cm. All 7 cases showed an MAML2 split pattern on fluorescence in situ hybridization analysis, and both RNA and whole-genome sequencing presented CRTC1::MAML2 translocation. All 7 cases showed a solid-predominant histology, and 3 cases displayed extracapsular extension. There were no other signs of high-grade histology and no recurrences or deaths occurred over a follow-up period of up to 79 months. We thus propose a unique trabecular variant of MEC that has atypical histologic and immunohistochemical features.
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Carcinoma Mucoepidermoide , Neoplasias de las Glándulas Salivales , Humanos , Femenino , Masculino , Carcinoma Mucoepidermoide/genética , Hibridación Fluorescente in Situ , Neoplasias de las Glándulas Salivales/genética , Factores de Transcripción/genética , Translocación Genética , Transactivadores/genéticaRESUMEN
MicroRNAs (miRNAs) are small non-coding RNAs known to play a significant role in the regulation of gene expression in various living organisms including fish. MiR-155 is known to enhance immunity in cells and several reports have demonstrated the antiviral properties of miR-155 in mammals. In this study, we investigated the antiviral role of miR-155 in Epithelioma papulosum cyprini (EPC) cells with viral hemorrhagic septicemia virus (VHSV) infection. EPC cells were transfected with miR-155 mimic and then infected with VHSV at different MOIs (0.01 and 0.001). The cytopathogenic effect (CPE) was observed at 0, 24, 48, and 72 h post infection (h.p.i). CPE progression appeared at 48 h.p.i in mock groups (VHSV only infected groups) and the VHSV infection group transfected with miR-155 inhibitors. On the other hand, the groups transfected with the miR-155 mimic did not show any CPE formation after infection with VHSV. The supernatant was collected at 24, 48 and 72 h.p.i., and the viral titers were measured by plaque assay. The viral titers increased at 48 and 72 h.p.i in groups infected only with VHSV. In contrast, the groups transfected with miR-155 did not show any increase in the virus titer and had a similar titer to 0 h.p.i. Furthermore, the real-time RT-PCR of immune gene expression showed upregulation of Mx1 and ISG15 at 0, 24, and 48 h.p.i in groups transfected with miR-155, while the genes were upregulated at 48 h.p.i in groups infected only with VHSV. Based on these results, miR-155 can induce the overexpression of type I interferon-related immune genes in EPCs and inhibit the viral replication of VHSV. Therefore, these results suggest that miR-155 could possess an antiviral effect against VHSV.
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Carcinoma , Enfermedades de los Peces , Septicemia Hemorrágica Viral , MicroARNs , Novirhabdovirus , Animales , Antivirales , Línea Celular Tumoral , MicroARNs/genética , MicroARNs/metabolismo , Novirhabdovirus/fisiología , Mamíferos/metabolismoRESUMEN
In the human body, proteins secreted into peripheral blood vessels are known as the secretome, and they represent the physiological or pathological status of cells. The unique response of cells to toxin exposure can be confirmed via secretome analysis, which can be used to discover toxic mechanisms or exposure markers. Alpha-amanitin (α-AMA) is the most widely studied amatoxin and inhibits transcription and protein synthesis by directly interacting with RNA polymerase II. However, secretory proteins released during hepatic failure caused by α-AMA have not been fully characterized. In this study, we analyzed the secretome of α-AMA-treated Huh-7 cells and mice using a comparative proteomics technique. Overall, 1440 and 208 proteins were quantified in cell media and mouse serum, respectively. Based on the bioinformatics results for the commonly downregulated proteins in cell media and mouse serum, we identified complement component 3 (C3) as a marker for α-AMA-induced hepatotoxicity. Through western blot in cell secretome and C3 ELISA assays in mouse serum, we validated α-AMA-induced downregulation of C3. In conclusion, using comparative proteomics and molecular biology techniques, we found that α-AMA-induced hepatotoxicity reduced C3 levels in the secretome. We expect that this study will aid in identifying new toxic mechanisms, therapeutic targets, and exposure markers of α-AMA-induced hepatotoxicity. Supplementary Information: The online version contains supplementary material available at 10.1007/s43188-022-00163-z.
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Physical shadow growth is a vacuum deposition technique that permits a wide variety of 3D-shaped nanoparticles and structures to be fabricated from a large library of materials. Recent advances in the control of the shadow effect at the nanoscale expand the scope of nanomaterials from spherical nanoparticles to complex 3D shaped hybrid nanoparticles and structures. In particular, plasmonically active nanomaterials can be engineered in their shape and material composition so that they exhibit unique physical and chemical properties. Here, the recent progress in the development of shadow growth techniques to realize hybrid plasmonic nanomaterials is discussed. The review describes how fabrication permits the material response to be engineered and highlights novel functions. Potential fields of application with a focus on photonic devices, biomedical, and chiral spectroscopic applications are discussed.
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The well-known hepatotoxicity mechanism resulting from alpha-amanitin (α-AMA) exposure arises from RNA polymerase II (RNAP II) inhibition. RNAP â ¡ inhibition occurs through the dysregulation of mRNA synthesis. However, the signaling pathways in hepatocytes that arise from α-AMA have not yet been fully elucidated. Here, we identified that the RAS/RAF/ERK signaling pathway was activated through quantitative phosphoproteomic and molecular biological analyses in Huh-7 cells. Bioinformatics analysis showed that α-AMA exposure increased protein phosphorylation in a time-dependent α-AMA exposure. In addition, phosphorylation increased not only the components of the ERK signaling pathway but also U2AF65 and SPF45, known splicing factors. Therefore, we propose a novel mechanism of α-AMA as follows. The RAS/RAF/ERK signaling pathway involved in aberrant splicing events is activated by α-AMA exposure followed by aberrant splicing events leading to cell death in Huh-7 cells.
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Alfa-Amanitina , ARN Polimerasa II , Alfa-Amanitina/farmacología , Sistema de Señalización de MAP Quinasas/fisiología , Fosforilación , Factores de Empalme de ARN , ARN MensajeroRESUMEN
The quantification of microRNAs (miRNAs) is important because the miRNA expression level is closely associated with the occurrence and development of diseases. Here, we report a simple nuclease protection transcription assay which combines nuclease protection assays and transcription-assisted light-up aptamer amplification for detecting miRNAs with great sensitivity.
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Aptámeros de Nucleótidos/genética , Técnicas Biosensibles , MicroARNs/genética , Técnicas de Amplificación de Ácido Nucleico , HumanosRESUMEN
A new strategy for developing versatile nanostructured surfaces utilizing the swelling of polymers in solvents is described. The self-stratified coating on 3D printed acrylonitrile-butadiene-styrene (ABS) copolymers with nanoparticles enables mechanically durable superhydrophobic characteristics. Unlike other methods, it was capable to produce superhydrophobicity on complex 3D structured surfaces. Mechanically durable superhydrophobic coatings that can withstand an abrasion cycle were obtained. Partial embedding of the nanoparticles into the ABS surface due to the swelling and self-stratification is considered as the reason for the increased mechanical strength of the coating. Utilizing this idea, the original concept of power-free physical sensors responding to changes in temperature, pressure, and surface tension was proposed.
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Alpha-amanitin (α-AMA) is a cyclic peptide and one of the most lethal mushroom amatoxins found in Amanita phalloides. α-AMA is known to cause hepatotoxicity through RNA polymerase II inhibition, which acts in RNA and DNA translocation. To investigate the toxic signature of α-AMA beyond known mechanisms, we used quantitative nanoflow liquid chromatography-tandem mass spectrometry analysis coupled with tandem mass tag labeling to examine proteome dynamics in Huh-7 human hepatoma cells treated with toxic concentrations of α-AMA. Among the 1828 proteins identified, we quantified 1563 proteins, which revealed that four subunits in the T-complex protein 1-ring complex protein decreased depending on the α-AMA concentration. We conducted bioinformatics analyses of the quantified proteins to characterize the toxic signature of α-AMA in hepatoma cells. This is the first report of global changes in proteome abundance with variations in α-AMA concentration, and our findings suggest a novel molecular regulation mechanism for hepatotoxicity.
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Alfa-Amanitina/toxicidad , Chaperonina con TCP-1/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Hígado/efectos de los fármacos , Intoxicación por Setas/metabolismo , Proteoma , Proteómica , Línea Celular Tumoral , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Cromatografía Liquida , Biología Computacional , Humanos , Hígado/metabolismo , Mapas de Interacción de Proteínas , Espectrometría de Masas en TándemRESUMEN
Vanishing white matter (VWM) disease is a genetic leukodystrophy leading to severe neurological disease and early death. VWM is caused by bi-allelic mutations in any of the five genes encoding the subunits of the eukaryotic translation factor 2B (EIF2B). Previous studies have attempted to investigate the molecular mechanism of VWN by constructing models for each subunit of EIF2B that causes VWM disease. The underlying molecular mechanisms of the way in which mutations in EIF2B3 result in VWM are largely unknown. Based on our recent results, we generated an eif2b3 knockout (eif2b3-/-) zebrafish model and performed quantitative proteomic analysis between the wild-type (WT) and eif2b3-/- zebrafish, and identified 25 differentially expressed proteins. Four proteins were significantly upregulated, and 21 proteins were significantly downregulated in eif2b3-/- zebrafish compared to WT. Lon protease and the neutral amino acid transporter SLC1A4 were significantly increased in eif2b3-/- zebrafish, and crystallin proteins were significantly decreased. The differential expression of proteins was confirmed by the evaluation of mRNA levels in eif2b3-/- zebrafish, using whole-mount in situ hybridization analysis. This study identified proteins which candidates as key regulators of the progression of VWN disease, using quantitative proteomic analysis in the first EIF2B3 animal model of VWN disease.
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Sistema de Transporte de Aminoácidos ASC/metabolismo , Leucoencefalopatías/metabolismo , Proteoma/metabolismo , Proteínas de Pez Cebra/metabolismo , Pez Cebra/metabolismo , Sistema de Transporte de Aminoácidos ASC/genética , Animales , Animales Modificados Genéticamente , Modelos Animales de Enfermedad , Factor 2B Eucariótico de Iniciación/deficiencia , Factor 2B Eucariótico de Iniciación/metabolismo , Humanos , Leucoencefalopatías/genética , Leucoencefalopatías/patología , Proteoma/genética , Proteómica , Pez Cebra/genética , Proteínas de Pez Cebra/genéticaRESUMEN
Obesity reduces adipocyte mitochondrial function, and expanding adipocyte oxidative capacity is an emerging strategy to improve systemic metabolism. Here, we report that serine/threonine-protein kinase 3 (STK3) and STK4 are key physiological suppressors of mitochondrial capacity in brown, beige and white adipose tissues. Levels of STK3 and STK4, kinases in the Hippo signalling pathway, are greater in white than brown adipose tissues, and levels in brown adipose tissue are suppressed by cold exposure and greatly elevated by surgical denervation. Genetic inactivation of Stk3 and Stk4 increases mitochondrial mass and function, stabilizes uncoupling protein 1 in beige adipose tissue and confers resistance to metabolic dysfunction induced by high-fat diet feeding. Mechanistically, STK3 and STK4 increase adipocyte mitophagy in part by regulating the phosphorylation and dimerization status of the mitophagy receptor BNIP3. STK3 and STK4 expression levels are elevated in human obesity, and pharmacological inhibition improves metabolic profiles in a mouse model of obesity, suggesting STK3 and STK4 as potential targets for treating obesity-related diseases.
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Adipocitos/metabolismo , Metabolismo Energético , Mitofagia , Proteínas Serina-Treonina Quinasas/metabolismo , Transducción de Señal , Tejido Adiposo Pardo/metabolismo , Tejido Adiposo Blanco/metabolismo , Animales , Línea Celular , Humanos , Péptidos y Proteínas de Señalización Intracelular , Ratones , Ratones Noqueados , Obesidad/prevención & control , Obesidad/terapia , Proteínas Serina-Treonina Quinasas/genética , Serina-Treonina Quinasa 3RESUMEN
Systemic toxicity, particularly, developmental defects of humidifier disinfectant chemicals that have caused lung injuries in Korean children, remains to be elucidated. This study evaluated the mechanisms of the adverse effects of 5-chloro-2-methyl-4-isothiazoline-3-one/2methyl-4-isothiazolin-3-one (CMIT/MIT), one of the main biocides of the Korean tragedy, and identify the most susceptible developmental stage when exposed in early life. To this end, the study was designed to analyze several endpoints (morphology, heart rate, behavior, global DNA methylation, gene expressions of DNA methyl-transferases (dnmts) and protein profiling) in exposed zebrafish (Danio rerio) embryos at various developmental stages. The results showed that CMIT/MIT exposure causes bent tail, pericardial edema, altered heart rates, global DNA hypermethylation and significant alterations in the locomotion behavior. Consistent with the morphological and physiological endpoints, proteomics profiling with bioinformatics analysis suggested that the suppression of cardiac muscle contractions and energy metabolism (oxidative phosphorylation) were possible pivotal underlying mechanisms of the CMIT/MIT mediated adverse effects. Briefly, multi-level endpoint analysis indicated the most susceptible window of exposure to be ≤ 6 hpf followed by ≤ 48 hpf for CMIT/MIT. These results could potentially be translated to a risk assessment of the developmental exposure effects to the humidifier disinfectants.
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Desinfectantes , Lesión Pulmonar , Animales , Niño , Desinfectantes/toxicidad , Embrión no Mamífero , Humanos , Proteómica , Pez CebraRESUMEN
BACKGROUND: Gamisoyo-San decoction (GSS), a traditional Chinese medicine, has been used to treat various gastrointestinal (GI) symptoms and diseases such as functional dyspepsia. The purpose of this study was to investigate the effect of GSS on GI motility functions in mice. METHODS: Percent intestinal transit rate (ITR%) and gastric emptying (GE) values were measured using Evans Blue and phenol red, respectively, in normal mice and in mice with experimentally induced GI motility dysfunction (GMD). RESULTS: In normal mice, GSS (0.01-1 g/kg) induced higher GE values than non-treated controls. Also, GSS could increase GE in loperamide-induced and cisplatin-induced GE delay models. In addition, GSS increased ITR% in a dose-dependent manner. Loperamide decreased ITR% and GSS recovered this loperamide-induced decrease in ITR%. To examine the effect of GSS on GMD, we used acetic acid (AA)-induced and streptozotocin (STZ)-induced mouse GMD models. The AA mouse model showed a significant decrease in ITR%. However, intragastric treatment with GSS significantly recovered this inhibition. Furthermore, STZ-induced diabetic mice showed a significant reduction in ITR%, which was also significantly inhibited by GSS. CONCLUSION: These results demonstrate that GSS can modulate bowel activity and that it could be used as a gastroprokinetic agent in the treatment of GI motility diseases.
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Medicamentos Herbarios Chinos/farmacología , Fármacos Gastrointestinales/farmacología , Enfermedades Gastrointestinales/tratamiento farmacológico , Tránsito Gastrointestinal/efectos de los fármacos , Ácido Acético/toxicidad , Animales , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/uso terapéutico , Fármacos Gastrointestinales/uso terapéutico , Enfermedades Gastrointestinales/inducido químicamente , Enfermedades Gastrointestinales/fisiopatología , Humanos , Masculino , Ratones , Ratones Endogámicos ICR , Estreptozocina/toxicidad , Resultado del TratamientoRESUMEN
BACKGROUND: The Gamisoyo-san (GSS) has been used for -improving the gastrointestinal (GI) symptoms. The purpose of this study was to investigate the effects of GSS, a traditional Chinese herbal medicine, on the pacemaker potentials of mouse small intestinal interstitial cells of Cajal (ICCs). METHODS: ICCs from the small intestines were dissociated and cultured. Whole-cell patch-clamp configuration was used to record pacemaker potentials and membrane currents. RESULTS: GSS depolarized ICC pacemaker potentials in a dose-dependent manner. Pretreatment with 4-diphenylacetoxypiperidinium iodide completely inhibited GSS-induced pacemaker potential depolarizations. Intracellular GDP-ß-S inhibited GSS-induced effects, and in the presence of U-73122, GSS-induced effects were inhibited. Also, GSS in the presence of a Ca2+-free solution or thapsigargin did not depolarize pacemaker potentials. However, in the presence of calphostin C, GSS slightly depolarized pacemaker potentials. Furthermore, GSS inhibited both transient receptor potential melastatin7 and Ca2+-activated Cl- channel (anoctamin1) currents. CONCLUSION: GSS depolarized pacemaker potentials of ICCs via G protein and muscarinic M3 receptor signaling pathways and through internal or external Ca2+-, phospholipase C-, and protein kinase C-dependent and transient receptor potential melastatin 7-, and anoctamin 1-independent pathways. The study shows that GSS may regulate GI tract motility, suggesting that GSS could be a basis for developing novel prokinetic agents for treating GI motility dysfunctions.