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1.
J Cell Mol Med ; 28(7): e18193, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38506205

RESUMEN

Colorectal cancer (CRC) liver metastasis, albeit a stage-IV disease, is completely curable by surgical resection in selected patients. In addressing the molecular basics of this phenomenon, differentially expressed genes at primary and liver metastatic sites were screened by RNA sequencing with the use of paraffin-embedded surgical specimens. Chemokine C-C motif ligand 1 (CCL1), a chemotactic factor for a ligand of the chemokine C-C motif receptor 8 (CCR8), was isolated as one of the differentially expressed genes. Histological analysis revealed that the number of CCL1-positive cells, mainly tumour associated macrophages (TAMs) located in the stroma of CRC, decreased significantly at liver metastatic sites, while the expression level of CCR8 on CRC remained unchanged. To explore the biological significance of the CCL1-CCR8 axis in CRC, CCR8-positive CRC cell line Colo320DM was used to assess the effect of the CCL1-CCR8 axis on major signalling pathways, epithelial mesenchymal transition induction and cell motility. Upon stimulation of recombinant CCL1 (rCCL1), phosphorylation of AKT was observed in Colo320DM cells; on the other hand, the corresponding significant increase in MMP-2 levels demonstrated by RT-qPCR was nullified by siRNA (siCCR8). In the scratch test, rCCL1 treatment significantly increased the motility of Colo320DM cells, which was similarly nullified by siCCR8. Thus, the activation of the CCL1-CCR8 axis is a positive regulator of CRC tumour progression. Reduced CCL1 expression of TAMs at liver metastatic sites may partly explain the unique slow tumour progression of CRC, thus providing for a grace period for radical resection of metastatic lesions.


Asunto(s)
Neoplasias Colorrectales , Hígado , Humanos , Quimiocina CCL1 , Ligandos , Hígado/metabolismo , Quimiocinas , Receptores de Quimiocina/metabolismo , Neoplasias Colorrectales/genética
2.
Genes Cells ; 28(5): 390-397, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-36861495

RESUMEN

We evaluated the efficacy of simultaneous multiple-gene knockout in human culture cells. By simple co-transfection of HeLa cells with a mixture of pX330-based targeting plasmids together with a puromycin resistance plasmid, followed by transient selection of puromycin-resistant cells, Cas9/single-guide RNA (sgRNA)-transduced polyclonal cell populations were selected and grown. Western blot analyses revealed co-transfection of up to seven targeting plasmids for p38α, p38ß, JNK1, JNK2, Mnk1, ERK1, and mLST8 genes, drastically reduced protein expression of these genes in the polyclonal population. Analyses of a randomly isolated group of 25 clones revealed knockout efficiencies for the seven targeted genes ranging between 68% and 100%, and in six clones (24%), all targeted genes were disrupted. Deep sequencing analyses of the individual target sites revealed that, in most cases, Cas9/sgRNA-induced nonhomologous end joining resulted in deletion or insertion of only a few base pairs at the break points. These results demonstrate that simple co-transfection-based simultaneous targeting offers an easy, rapid, and efficient method to generate multiplex gene-knockout cell lines.


Asunto(s)
Sistemas CRISPR-Cas , Edición Génica , Humanos , Sistemas CRISPR-Cas/genética , Células HeLa , Técnicas de Inactivación de Genes , Plásmidos , Transfección , Edición Génica/métodos
3.
Surg Today ; 54(2): 205-209, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-37516666

RESUMEN

We reported previously that a large vertical interval between the hepatic segment of the inferior vena cava (IVC) and right atrium (RA), referred to as the IVC-RA gap, was associated with more intraoperative bleeding during hemi-hepatectomy. We conducted a computational fluid dynamics (CFD) study to clarify the impact of fluid dynamics resulting from morphologic variations around the liver. The subjects were 10 patients/donors with a large IVC-RA gap and 10 patients/donors with a small IVC-RA gap. Three-dimensional reconstructions of the IVC and hepatic vessels were created from CT images for the CFD study. Median pressure in the middle hepatic vein was significantly higher in the large-gap group than in the small-gap group (P = 0.008). Differences in hepatic vein pressure caused by morphologic variation in the IVC might be one of the mechanisms of intraoperative bleeding from the hepatic veins.


Asunto(s)
Venas Hepáticas , Vena Cava Inferior , Humanos , Vena Cava Inferior/anatomía & histología , Venas Hepáticas/anatomía & histología , Hidrodinámica , Hígado/diagnóstico por imagen , Hepatectomía/métodos
4.
Surg Today ; 2024 Mar 13.
Artículo en Inglés | MEDLINE | ID: mdl-38478124

RESUMEN

PURPOSE: Post-transplant biliary stricture (PBS) is a common and important complication following orthotopic liver transplantation (LT). This study clarified the incidence of PBS and identified its risk factors. METHODS: We retrospectively reviewed the medical records of 67 patients who underwent living-donor LT (LDLT) at our institute between June 2010 and July 2022 and analyzed their clinical characteristics, prognosis, and risk factors for PBS. RESULTS: Of the 67 patients, 26 (38.8%) developed PBS during the observation period. Multivariate analyses revealed the following independent risk factors for PBS formation: increased red cell transfusion volume per body weight (> 0.2 U/kg; hazard ratio [HR], 3.8; P = 0.002), increased portal vein pressure (PVP) at the end of LT (> 16 mmHg; HR, 2.88; P = 0.032), postoperative biliary leakage (HR, 4.58; P = 0.014), and prolonged warm ischemia time (WIT) (> 48 min; HR, 4.53; P = 0.008). In patients with PBS, the cumulative incidence of becoming stent free was significantly higher in patients with a WIT ≤ 48 min than in those with a WIT > 48 min (P = 0.038). CONCLUSION: Prolonged WIT is associated with intractable PBS following LDLT.

5.
Histochem Cell Biol ; 160(5): 407-418, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37532885

RESUMEN

The liver is known to possess remarkable regenerative potential, but persistent inflammation or severe acute injury can lead to liver fibrosis and incomplete regeneration, ultimately resulting in liver failure. Recent studies have shown that the axis of two types of CXCL12 receptors, CXCR4 and CXCR7, plays a crucial role in liver fibrosis and regeneration. The present study aimed to investigate the regulatory factors involved in CXCR4 expression in injured liver. Immunohistochemical screening of liver tissue samples collected during liver transplantation revealed a reciprocal expression pattern between CXCR4 and MeCP2. An in vitro system involving cultured cell lines and H2O2 treatment was established to study the impact of oxidative stress on signaling pathways and epigenetic alterations that affect CXCR4 mRNA expression. Operating through distinct signaling pathways, H2O2 treatment induced a dose-dependent increase in CXCR4 expression in both hepatocyte- and intrahepatic cholangiocyte-derived cells. Treatment of the cells with trichostatin and azacytidine modulated CXCR4 expression in hepatocytes by modifying the methylation status of CpG dinucleotides located in a pair of TA repeats adjacent to the TATA box of the CXCR4 gene promoter. Only MeCP2 bound to oligonucleotides representing the TATA box region when the cytosine residues within the sequence were methylated, as revealed by electrophoretic mobility shift assay (EMSA). Methylation-specific PCR analysis of microdissected samples revealed a correlation between the loss of CpG methylation and the upregulation of CXCR4 in injured hepatocytes, replicating the findings from the in vitro study. Besides the conventional MEK/ERK and NF-κB signaling pathways that activate CXCR4 in intrahepatic cholangiocytes, the unique epigenetic modifications observed in hepatocytes might also contribute to a shift in the CXCR4-CXCR7 balance towards CXCR4, leading to irreversible liver injury and fibrosis. This study highlights the importance of epigenetic modifications in regulating CXCR4 expression in liver injury and fibrosis.


Asunto(s)
Peróxido de Hidrógeno , Receptores CXCR4 , Humanos , Receptores CXCR4/genética , Receptores CXCR4/metabolismo , Hepatocitos/metabolismo , Cirrosis Hepática , Regiones Promotoras Genéticas , Desmetilación , Expresión Génica , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Quimiocina CXCL12/farmacología
6.
World J Surg ; 47(1): 260-268, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-36261603

RESUMEN

BACKGROUND: Incisional hernia (IH) is a common surgical complication, with an incidence of 6-31% following major abdominal surgery. This study aimed to investigate the impact of intramuscular adipose tissue content (IMAC) on the incidence of IH in patients who underwent hepatic resection. METHODS: Data of 205 patients who underwent open hepatic resection between 2007 and 2019 at Ehime University Hospital were retrospectively analyzed. Patient characteristics, perioperative findings, and body composition were compared between patients with IH and those without IH. The quantity and quality of skeletal muscle, calculated as skeletal muscle index and IMAC, were evaluated using preoperative computerized tomography images. RESULTS: Forty (19.5%) patients were diagnosed with IH. The cumulative incidence rates were 15.6% at 1 year and 19.6% at 3 years. On univariate analysis, body mass index, areas of subcutaneous and visceral fat, and IMAC were significantly higher in the IH group than in the non-IH group (p = 0.0023, 0.0070, 0.0047, and 0.0080, respectively). No significant difference in skeletal muscle index was found between the groups (p = 0.3548). The incidence of diabetes mellitus, intraoperative transfusion, and postoperative wound infection was significantly higher in the IH group than in the non-IH group (p = 0.0361, 0.0078, and 0.0299, respectively). On multivariate analysis, a high IMAC and wound infection were independent risk factors for IH (adjusted odds ratio, 2.83 and 4.52, respectively; p = 0.0152 and 0.0164, respectively). CONCLUSION: IMAC can predict the incidence of IH in patients undergoing hepatic resection.


Asunto(s)
Hernia Incisional , Humanos , Hernia Incisional/diagnóstico por imagen , Hernia Incisional/epidemiología , Hernia Incisional/etiología , Estudios Retrospectivos , Tejido Adiposo
7.
Cytopathology ; 34(4): 308-317, 2023 07.
Artículo en Inglés | MEDLINE | ID: mdl-37051774

RESUMEN

OBJECTIVE: Artificial intelligence (AI)-based cytopathology studies conducted using deep learning have enabled cell detection and classification. Liquid-based cytology (LBC) has facilitated the standardisation of specimen preparation; however, cytomorphology varies according to the LBC processing technique used. In this study, we elucidated the relationship between two LBC techniques and cell detection and classification using a deep learning model. METHODS: Cytological specimens were prepared using the ThinPrep and SurePath methods. The accuracy of cell detection and cell classification was examined using the one- and five-cell models, which were trained with one and five cell types, respectively. RESULTS: When the same LBC processing techniques were used for the training and detection preparations, the cell detection and classification rates were high. The model trained on ThinPrep preparations was more accurate than that trained on SurePath. When the preparation types used for training and detection were different, the accuracy of cell detection and classification was significantly reduced (P < 0.01). The model trained on both ThinPrep and SurePath preparations exhibited slightly reduced cell detection and classification rates but was highly accurate. CONCLUSIONS: For the two LBC processing techniques, cytomorphology varied according to cell type; this difference affects the accuracy of cell detection and classification by deep learning. Therefore, for highly accurate cell detection and classification using AI, the same processing technique must be used for both training and detection. Our assessment also suggests that a deep learning model should be constructed using specimens prepared via a variety of processing techniques to construct a globally applicable AI model.


Asunto(s)
Inteligencia Artificial , Aprendizaje Profundo , Humanos , Técnicas Citológicas/métodos , Citodiagnóstico/métodos
8.
Tohoku J Exp Med ; 260(1): 1-11, 2023 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-36823184

RESUMEN

Numerous studies have investigated the impact of the coronavirus disease 2019 (COVID-19) pandemic on mental health among university students within a year of its onset, but few have examined the impact of a prolonged pandemic on university life. This study aimed to evaluate the impact of the COVID-19 pandemic on the mental health of students in a large university community. Online questionnaire surveys were conducted on students from March 24 to April 14 (first survey, n = 3,357) and December 2-23, 2021 (second survey, n = 2,604). The questionnaires included items on demographic data, living conditions, and mental health status as measured using the Patient Health Questionnaire-9 for depressive symptoms and the Generalized Anxiety Disorder-7 scale for anxiety symptoms. The results showed that, compared with undergraduate students, graduate students, except those in Medicine, Dentistry, and Pharmaceutical Science courses, had more anxiety symptoms. Furthermore, among undergraduate students, depressive and anxiety symptoms were significantly higher in fourth- than in first-year students. Logistic regression analyses of data from both surveys revealed the seven risk factors associated with depressive or anxiety symptoms that affected the mental health of university students throughout the COVID-19 pandemic: 1) female or nonbinary gender, 2) graduate student, 3) quarantine experience due to COVID-19, 4) isolation from friends and acquaintances, 5) disorganized pattern of daily life, 6) worse financial situation, and 7) no availability of consultations regarding health, life, and finances. These findings suggest that mental health measures for university students need to be designed specific to each course.


Asunto(s)
COVID-19 , Humanos , Femenino , COVID-19/epidemiología , Pandemias , Estudios Transversales , Salud Mental , SARS-CoV-2 , Universidades , Japón/epidemiología , Depresión/psicología , Ansiedad/epidemiología , Ansiedad/etiología , Estudiantes
9.
Pancreatology ; 22(5): 651-655, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35487869

RESUMEN

BACKGROUND: /Objectives: Postoperative pancreatic fistula (POPF) is a serious complication after pancreaticoduodenectomy (PD). Thus, identification of the risk factors for POPF is urgently needed. In this study, we aimed to identify whether arterial lactate (LCT) levels following PD might be a marker of the potential risk of POPF. METHODS: Between September 2009 and December 2020, 151 patients who underwent elective PD were retrospectively enrolled. Patient characteristics, perioperative clinicopathological variables, postoperative blood biochemistry data were analyzed in univariable and multivariable analyses. Pancreatic fistula of Grade B and C was considered as POPF. RESULTS: Patients were divided into the POPF group (n = 33, 21.9%) and non-POPF group (n = 118, 78.1%). Higher body mass index (p = 0.017), increased estimated blood loss (p = 0.047), soft textured pancreas (p = 0.007), smaller main pancreatic duct (p = 0.016), higher LCT levels (p < 0.001), higher aspartate aminotransferase levels (p = 0.023) and higher procalcitonin levels (p = 0.024) were significantly associated with POPF. Receiver operating characteristic curve analysis revealed that 2.1 mmol/L was the optimal cut-off value of LCT (sensitivity = 78.8%, specificity = 61.2%) for predicting POPF occurrence. Univariate and multivariate analyses confirmed that an LCT of ≥2.1 mmol/L was independently associated with the risk of POPF following PD (odds ratio = 6.78, 95% confidence interval = 2.22-20.74; p = 0.001). CONCLUSIONS: Higher LCT is a predictive marker for POPF following PD.


Asunto(s)
Fístula Pancreática , Pancreaticoduodenectomía , Humanos , Lactatos , Páncreas/patología , Fístula Pancreática/diagnóstico , Fístula Pancreática/epidemiología , Fístula Pancreática/etiología , Pancreaticoduodenectomía/efectos adversos , Complicaciones Posoperatorias/etiología , Estudios Retrospectivos , Factores de Riesgo
10.
Zoolog Sci ; 39(6): 562-569, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36495491

RESUMEN

Juvenile hormone (JH) has two major functions in insects, i.e., suppression of metamorphosis in the larval or nymphal stage and promotion of reproduction in the adult stage. Krüppel homolog 1 (Kr-h1), a C2H2 zinc-finger type transcription factor, is reported to act downstream of the JH receptor complex. In the present study, the function of Kr-h1 was examined in adults and nymphs of Riptortus pedestris by RNA interference (RNAi). After injection of adults with dsRNA of Kr-h1, the expression level of Kr-h1 was significantly decreased in the abdomen. Kr-h1 dsRNA-injection resulted in a lower proportion of individuals with developed ovaries, but the difference was not statistically significant. The transcript levels of cyanoprotein-α and vitellogenin-1, which are JH-inducible genes encoding yolk proteins, were not affected in the abdomen by Kr-h1 knockdown. Kr-h1 dsRNA-injection was effective for suppression of Kr-h1 expression in nymphs. Some Kr-h1 dsRNA-injected fifth (final) instar nymphs had morphological defects in the wing bud. Moreover, they had several adult morphological features, including ocelli in the head, connexivum in the abdomen, coloring of the dorsal abdomen, and genitals. The nymphs possessing adult features did not emerge as adults during 1 month. These results demonstrated that Kr-h1 is necessary for maintaining nymphal characters in R. pedestris. The function of Kr-h1 in ovarian development remains unclear in R. pedestris.


Asunto(s)
Heterópteros , Hormonas Juveniles , Animales , Hormonas Juveniles/metabolismo , Factores de Transcripción/genética , Factores de Transcripción de Tipo Kruppel/genética , Factores de Transcripción de Tipo Kruppel/metabolismo , Regulación del Desarrollo de la Expresión Génica , Metamorfosis Biológica , Heterópteros/genética , Interferencia de ARN , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo
11.
Langenbecks Arch Surg ; 407(4): 1585-1594, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-34997276

RESUMEN

PURPOSE: The aim of the present study on living donor liver transplantation (LDLT) using a right-lobe graft without the middle hepatic vein (MHV) was to investigate the clinical impact of MHV tributary reconstruction using our criteria and techniques. METHODS: The medical records of 40 patients who underwent adult LDLT using a right-lobe graft without the MHV between April 2008 and December 2020 were retrospectively reviewed. In this cohort, the criterion for MHV tributary reconstruction was estimated drainage volume of each MHV tributary greater than 100 mL. The drainage vein of segment 8 (V8) was reconstructed as the common orifice of the right hepatic vein and V8 using a venous patch graft, and that of segment 5 was reconstructed using artificial vascular grafts. The outcomes were compared between the groups with and without MHV tributary reconstruction. Factors associated with postoperative massive ascites were also investigated. RESULTS: Twenty patients underwent MHV tributary reconstruction. There were no significant differences in the amount of postoperative ascites, Clavien-Dindo classification ≥ III postoperative complications, and 90-day in-hospital mortality between the groups (P = 0.678, P = 1.000, and P = 0.244, respectively). On multivariate analyses, a low-estimated functional graft-to-recipient weight ratio, which was calculated using estimated graft volume minus the territory of MHV tributaries that was not reconstructed, was identified as an independent predictor of postoperative massive ascites (odds ratio, 40.479; 95% confidence interval, 3.823-428.622). CONCLUSION: The present study suggests that selective MHV tributary reconstruction might be useful for achieving successful graft function.


Asunto(s)
Trasplante de Hígado , Donadores Vivos , Adulto , Ascitis , Venas Hepáticas/cirugía , Humanos , Hígado , Trasplante de Hígado/métodos , Estudios Retrospectivos
12.
Surg Today ; 52(4): 721-725, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-34853880

RESUMEN

The perioperative management and technical details of laparoscopic clamp-crushing enucleation for low-malignant-potential pancreatic neuroendocrine neoplasms (PNENs) located close to the main pancreatic duct (MPD) in the body/tail of the pancreas using a perioperative MPD stent are reported. The procedure was performed in two patients with PNEN (13 and 10 mm in diameter) in the body/tail of the pancreas. A naso-pancreatic stent (NPS) was placed preoperatively in both patients. Resection was performed using Maryland-type bipolar forceps. The surgical duration was 139 and 55 min, and the estimated blood loss was 5 and 0 mL, respectively. One patient was discharged uneventfully on postoperative day (POD) 12. The other patient developed a grade B pancreatic fistula, but was discharged on POD 22. Laparoscopic clamp-crushing enucleation with an NPS might be a viable treatment option for tumors located close to the MPD.


Asunto(s)
Laparoscopía , Neoplasias Pancreáticas , Humanos , Laparoscopía/métodos , Páncreas/cirugía , Pancreatectomía/métodos , Conductos Pancreáticos/patología , Conductos Pancreáticos/cirugía , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/cirugía , Stents
13.
Water Sci Technol ; 85(10): 2827-2839, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-35638790

RESUMEN

To increase the adsorption capability of Hg2+ from aqueous media, we prepared sodium-type fine zeolite grains with various particle sizes (denoted as ZE1, ZE2 and ZE3). The particle sizes of ZE1, ZE2 and ZE3 were 16.363 ± 0.365, 1.454 ± 0.357 and 0.607 ± 0.377 µm, respectively. Moreover, the CEC, specific surface area and pore volume were in the order ZE1 (42 mmol/g and 23.5 m2/g) < ZE2 (72 mmol/g and 67.1 m2/g) < ZE3 (135 mmol/g and 176.6 m2/g). Subsequently, the Hg2+ adsorption capability was investigated. The performance of tested agents on Hg2+ adsorbed was in the order ZE1 (5.0 mg/g) < ZE2 (9.4 mg/g) < ZE3 (20.2 mg/g). It was concluded that fine crystalline zeolite was important in enhancing the adsorption capability of Hg2+. In addition, the mechanism of adsorption of Hg2+ on the ZE samples was evaluated. Our results suggested that Hg2+ was exchanged with sodium ions in the interlayers of ZE samples with correlation coefficients of 0.966-0.979. Our findings revealed that these ZE samples constitute potential agents for the adsorption of Hg2+ from aqueous media.


Asunto(s)
Mercurio , Zeolitas , Adsorción , Iones , Mercurio/química , Sodio , Zeolitas/química
14.
Gan To Kagaku Ryoho ; 49(12): 1365-1367, 2022 Dec.
Artículo en Japonés | MEDLINE | ID: mdl-36539251

RESUMEN

A 57-year-old man was treated with lenvatinib for unresectable hepatocellular carcinoma(HCC). Thereafter, the tumor marker levels decreased, and the tumor became resectable. The patient underwent portal vein embolization followed by laparoscopic extended left lobectomy. The patient's postoperative course was uneventful, and the tumor marker levels remained within the normal range. No recurrence was observed 3 months after surgery. In recent years, the use of systemic chemotherapy with drugs, such as lenvatinib, followed by conversion surgery has been reported in some cases of unresectable HCC. The present case reports successful conversion surgery following lenvatinib treatment.


Asunto(s)
Carcinoma Hepatocelular , Neoplasias Hepáticas , Quinolinas , Masculino , Humanos , Persona de Mediana Edad , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/cirugía , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/cirugía , Compuestos de Fenilurea/uso terapéutico , Quinolinas/uso terapéutico , Biomarcadores de Tumor
15.
Anal Chem ; 92(24): 16058-16065, 2020 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-33172270

RESUMEN

Thermal ionization mass spectrometry (TIMS) was used to directly quantify an ultratrace of radioactive 90Sr in microliter droplet samples. No chemical separation was required in removing isobaric interferences on M = 90 such as 90Zr and organic molecules in the mass spectrum because the difference in evaporation and ionization (emission) temperature among organic molecules, Zr and Sr, allows us to control the emission manner and significantly suppress the isobaric interferences. Direct quantification was achieved by improving the intercalibration of Faraday cups and ion counting in an isotope dilution (ID) method. Furthermore, the use of a total evaporation method (TE) enhanced the detection efficiency by the complete collection of the 90Sr ion beam from the samples and minimized the complexity of the fractionation effect in the isotope ratio calculation. In this study, 1 fg of 90Sr (equal to activity of 5 millibecquerel (mBq)) in a very low-volume sample with 108 times greater isobaric interference from 90Zr was successfully measured using the proposed ID-TE-TIMS method. The limit of detection was 0.029 fg (equal to 0.15 mBq) without any preconcentration. To demonstrate the wide usability of this method, low-volume samples of tears, eyelashes, saliva, environmental standards, and water samples (i.e., seawater and ground water) were analyzed within 1 h. The relationship of the measured values between this ID-TE-TIMS method and a radiometric analysis was shown to have good linearity.

16.
Doc Ophthalmol ; 139(1): 33-44, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-30937682

RESUMEN

PURPOSE: To investigate the relationship between adaptation time and the parameters of electroretinography (ERG) and pupillography in healthy subjects. METHODS: Forty-six eyes of 23 healthy women (mean age 21.7 years) were enrolled. ERG and pupillography were tested in each of the right and left 23 eyes, respectively. ERG with a skin electrode was used to determine amplitude and implicit time by the records of rod-, flash-, cone-, and flicker-responses with white light (0.01-30 cd s/m2). Infrared pupillography was used to record the pupillary light reflex to 1-s stimulation of red light (100 cd/m2). Cone- and flicker- (rod-, flash- and pupil) responses were recorded after light (dark) adaptation at 1, 5, 10, 15, and 20 min. RESULTS: Amplitude (µV) was significantly different between 1 min and ≥ 5 or ≥ 10 min after adaptation in b-wave of cone- or rod-response, respectively. Implicit time (ms) differed significantly between 1 min and ≥ 5 min after adaptation with b-wave of cone- and rod-response. There were significant differences between 1 min and ≥ 10 or ≥ 5 min after dark adaptation in parameter of minimum pupil diameter (mm) or constriction rate (%), respectively. CONCLUSIONS: Cone-driven ERG can be recorded, even in 5 min of light adaptation time without any special light condition, whereas rod-driven ERG and pupillary response results can be obtained in 10 min or longer of dark adaptation time in complete darkness.


Asunto(s)
Adaptación Ocular/fisiología , Células Fotorreceptoras de Vertebrados/fisiología , Pupila/fisiología , Reflejo Pupilar/fisiología , Adulto , Electrorretinografía/métodos , Femenino , Voluntarios Sanos , Humanos , Estimulación Luminosa , Factores de Tiempo , Adulto Joven
17.
Biochem J ; 474(10): 1669-1687, 2017 05 04.
Artículo en Inglés | MEDLINE | ID: mdl-28298474

RESUMEN

Although tumor protein D52 (TPD52) family proteins were first identified nearly 20 years ago, their molecular regulatory mechanisms remain unclear. Therefore, we investigated the post-transcriptional regulation of TPD52 family genes. An RNA immunoprecipitation (RIP) assay showed the potential binding ability of TPD52 family mRNAs to several RNA-binding proteins, and an RNA degradation assay revealed that TPD52 is subject to more prominent post-transcriptional regulation than are TPD53 and TPD54. We subsequently focused on the 3'-untranslated region (3'-UTR) of TPD52 as a cis-acting element in post-transcriptional gene regulation. Several deletion mutants of the 3'-UTR of TPD52 mRNA were constructed and ligated to the 3'-end of a reporter green fluorescence protein gene. An RNA degradation assay revealed that a minimal cis-acting region, located in the 78-280 region of the 5'-proximal region of the 3'-UTR, stabilized the reporter mRNA. Biotin pull-down and RIP assays revealed specific binding of the region to T-cell intracellular antigen 1 (TIA-1) and TIA-1-related protein (TIAR). Knockdown of TIA-1/TIAR decreased not only the expression, but also the stability of TPD52 mRNA; it also decreased the expression and stability of the reporter gene ligated to the 3'-end of the 78-280 fragment. Stimulation of transforming growth factor-ß and epidermal growth factor decreased the binding ability of these factors, resulting in decreased mRNA stability. These results indicate that the 78-280 fragment and TIA-1/TIAR concordantly contribute to mRNA stability as a cis-acting element and trans-acting factor(s), respectively. Thus, we here report the specific interactions between these elements in the post-transcriptional regulation of the TPD52 gene.


Asunto(s)
Regulación Neoplásica de la Expresión Génica , Proteínas de Neoplasias/metabolismo , Proteínas de Unión a Poli(A)/metabolismo , ARN Mensajero/metabolismo , ARN Neoplásico/metabolismo , Proteínas de Unión al ARN/metabolismo , Elementos de Respuesta , Regiones no Traducidas 3' , Línea Celular Tumoral , Células Cultivadas , Técnicas de Silenciamiento del Gen , Genes Reporteros , Humanos , Inmunoprecipitación , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/química , Proteínas de Neoplasias/genética , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/química , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/metabolismo , Proteínas de Unión a Poli(A)/antagonistas & inhibidores , Proteínas de Unión a Poli(A)/genética , Interferencia de ARN , Estabilidad del ARN , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/química , ARN Neoplásico/antagonistas & inhibidores , ARN Neoplásico/química , Proteínas de Unión al ARN/antagonistas & inhibidores , Proteínas de Unión al ARN/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Eliminación de Secuencia , Antígeno Intracelular 1 de las Células T
18.
Biosci Biotechnol Biochem ; 81(3): 534-540, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28051915

RESUMEN

To develop new whitening agents from natural products, we screened 80 compounds derived from crude drugs in Kampo medicine in a melanin synthesis inhibition assay using murine B16 melanoma cells. The screen revealed that treatment with alisol B, a triterpene from Alismatis rhizoma, significantly decreased both melanin content and cellular tyrosinase activity in B16 cells. However, alisol B did not directly inhibit mushroom tyrosinase activity in vitro. Therefore, we investigated the mechanism underlying the inhibitory effect of alisol B on melanogenesis. Alisol B suppressed mRNA induction of tyrosinase and its transcription factor, microphthalmia-associated transcription factor (MITF). Furthermore, alisol B reduced the phosphorylation of CREB and maintained the activation of ERK1/2. These results suggest that the reduction in melanin production by alisol B is due to the downregulation of MITF through the suppression of CREB and activation of ERK and that alisol B may be useful as a new whitening agent.


Asunto(s)
Alisma/química , Colestenonas/farmacología , Melaninas/biosíntesis , Melanoma Experimental/metabolismo , Preparaciones para Aclaramiento de la Piel/farmacología , Animales , Línea Celular Tumoral , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Evaluación Preclínica de Medicamentos/métodos , Melaninas/antagonistas & inhibidores , Melanoma Experimental/tratamiento farmacológico , Melanoma Experimental/patología , Ratones , Factor de Transcripción Asociado a Microftalmía/genética , Factor de Transcripción Asociado a Microftalmía/metabolismo , Monofenol Monooxigenasa/genética , Monofenol Monooxigenasa/metabolismo , Fosforilación/efectos de los fármacos , Rizoma/química , Transducción de Señal/efectos de los fármacos
19.
Planta Med ; 83(9): 812-818, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28158891

RESUMEN

The study of the chemical constituents of branches and twigs of Cratoxylum cochinchinense collected in Singapore led to the isolation and structural elucidation of four new xanthones, named cratoxanthone A (1), B (2), C (3), and D (4), together with six known xanthones (5-10) and one known dihydroanthracenone (11). Eight xanthones (including 1 and 2) and 11 were tested for their antiproliferative activity in three human carcinoma cell lines (lung adenocarcinoma A549, colorectal carcinoma Colo205, and epidermoid carcinoma KB) and a human acute lymphoblastic leukemia B cell line (NALM-6), and the mitochondrial membrane potential was determined in KB cells. New xanthones 1 and 2 attenuated NALM-6 cell proliferation with IC50 values of 17.78 and 8.27 µM, respectively. Furthermore, KB cells treated with these compounds had significantly decreased mitochondrial membrane potentials. Notably, the proliferation of A549 cells was specifically inhibited by 11, but not the xanthones.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Proliferación Celular/efectos de los fármacos , Clusiaceae/química , Extractos Vegetales/farmacología , Xantonas/farmacología , Antineoplásicos Fitogénicos/aislamiento & purificación , Línea Celular Tumoral , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Brotes de la Planta/química , Singapur , Xantonas/aislamiento & purificación
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