RESUMEN
High-speed atomic force microscopy (HS-AFM) is an indispensable technique in the field of biology owing to its imaging capability with high spatiotemporal resolution. Furthermore, recent developments established tip-scan stand-alone HS-AFM combined with an optical microscope, drastically improving its versatility. It has considerable potential to contribute to not only biology but also various research fields. A great candidate is a photoactive material, such as an azo-polymer, which is important for optical applications because of its unique nanoscale motion under light irradiation. Here, we demonstrate the in situ observation of nanoscale azo-polymer motion by combining tip-scan HS-AFM with an optical system, allowing HS-AFM observations precisely aligned with a focused laser position. We observed the dynamic evolution of unique morphologies in azo-polymer films. Moreover, real-time topographic line profile analyses facilitated precise investigations of the morphological changes. This important demonstration would pave the way for the application of HS-AFM in a wide range of research fields.
RESUMEN
Increasing evidence indicates that enhanced peripheral carbonyl stress markers exist in subtype of schizophrenia, although it may not be the primary cause. This study aimed to investigate whether plasma concentrations of methylglyoxal, 3-deoxy-glucosone, and glyoxal, which are reactive intermediates of protein metabolism in carbonyl stress, are changed in patients with schizophrenia and can function as potential biomarkers for schizophrenia with enhanced carbonyl stress. Plasma concentrations of these di-carbonyls were simultaneously estimated in 40 patients with schizophrenia and 40 healthy controls. As a result, no statistically significant differences were observed in mean plasma concentrations of three di-carbonyls between patients and controls. However, a remarkable increase in methylglyoxal concentrations was observed in four patients but not in controls. This increase was not found with regard to 3-deoxyglucosone and glyoxal both of patients and controls. Our correlation analysis showed that both the plasma methylglyoxal and glyoxal concentrations were significantly correlated with 3-deoxyglucosone concentrations in 40 patients and 40 controls. However, the plasma methylglyoxal concentrations did not show any significant correlation with the glyoxal concentrations in the patients or the controls. In four patients with extremely high methylglyoxal levels, the plasma methylglyoxal and glyoxal concentrations were not correlated to the 3-deoxyglucosone concentrations. Methylglyoxal is a physiological substrate of the glyoxalase system, and the accelerated accumulation of this compound lowers the glyoxalase I activity. These results suggested that this increase in four patients with high methylglyoxal levels may indicate the presence of a subtype of chronic schizophrenia that is associated with enhanced carbonyl stress.
Asunto(s)
Piruvaldehído/sangre , Esquizofrenia/sangre , Adulto , Biomarcadores/sangre , Estudios de Casos y Controles , Enfermedad Crónica , Desoxiglucosa/análogos & derivados , Desoxiglucosa/sangre , Femenino , Glioxal/sangre , Hospitalización , Humanos , Masculino , Persona de Mediana Edad , Estrés FisiológicoRESUMEN
Recent studies have revealed that newly emerging transformed cells are often apically extruded from epithelial tissues. During this process, normal epithelial cells can recognize and actively eliminate transformed cells, a process called epithelial defence against cancer (EDAC). Here, we show that mitochondrial membrane potential is diminished in RasV12-transformed cells when they are surrounded by normal cells. In addition, glucose uptake is elevated, leading to higher lactate production. The mitochondrial dysfunction is driven by upregulation of pyruvate dehydrogenase kinase 4 (PDK4), which positively regulates elimination of RasV12-transformed cells. Furthermore, EDAC from the surrounding normal cells, involving filamin, drives the Warburg-effect-like metabolic alteration. Moreover, using a cell-competition mouse model, we demonstrate that PDK-mediated metabolic changes promote the elimination of RasV12-transformed cells from intestinal epithelia. These data indicate that non-cell-autonomous metabolic modulation is a crucial regulator for cell competition, shedding light on the unexplored events at the initial stage of carcinogenesis.
Asunto(s)
Comunicación Celular , Transformación Celular Neoplásica/metabolismo , Metabolismo Energético , Células Epiteliales/metabolismo , Animales , Línea Celular Transformada , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/patología , Técnicas de Cocultivo , Proteínas del Citoesqueleto/genética , Proteínas del Citoesqueleto/metabolismo , Perros , Femenino , Genes ras , Glucosa/metabolismo , Glucólisis , Ácido Láctico/metabolismo , Células de Riñón Canino Madin Darby , Masculino , Potencial de la Membrana Mitocondrial , Ratones Endogámicos C57BL , Ratones Transgénicos , Mitocondrias/metabolismo , Proteínas Quinasas/genética , Proteínas Quinasas/metabolismo , Interferencia de ARN , Transducción de Señal , Técnicas de Cultivo de Tejidos , TransfecciónRESUMEN
We encountered 2 patients with unresectable advanced gallbladder cancer whose performance status improved within a short time following the administration of gemcitabine and cisplatin. Both patients were able to maintain a good QOL while continuing treatment.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Vesícula Biliar/tratamiento farmacológico , Anciano , Cisplatino/administración & dosificación , Desoxicitidina/administración & dosificación , Desoxicitidina/análogos & derivados , Resultado Fatal , Femenino , Humanos , Masculino , Persona de Mediana Edad , Resultado del Tratamiento , GemcitabinaRESUMEN
Mutations in the adenomatous polyposis coli (APC) gene contribute to the progression of colorectal tumorigenesis. Despite the importance, few studies regarding the localization of this protein on surgically resected human colorectal cancer specimens using immunohistochemistry have been reported so far because of the unavailability of the antibodies for this use. The goal of this study has been to provide the APC protein expression and to validate the APC molecular studies. We took advantage of an immunohistochemistry procedure of applying the unique detergent-mediated antigen retrieval technique to frozen sections and examined the expressions of one amino (N)-terminal (AC4) and one carboxy (C)-terminal APC antibody (HG2). Further, we compared the stainings of APC antibodies with those of the E-cadherin antibody using a quantitative image analysis. E-cadherin is a critical morphogenetic regulator during embryogenesis and recent evidence strongly suggests that downregulation of E-cadherin expression in cancers is associated with a high rate of invasion and metastasis. The analysis indicated statistically that normal epithelia showed stronger staining than cancer cells ( P<0.05). Further, in normal epithelia, the amino (N)-terminal APC antibody (AC4) showed a positive correlation with another carboxy (C)-terminal APC antibody (HG2). E-cadherin showed no positive correlation with other APCs in either the normal epithelia or cancer cells. This study verified reduced expressions of APCs and E-cadherin proteins in colorectal cancer cells. This suggests that the normal APC and E-cadherin protein expressions in benign epithelium are progressively and independently lost in the sporadic colorectal cancers.
Asunto(s)
Proteína de la Poliposis Adenomatosa del Colon/análisis , Cadherinas/análisis , Neoplasias Colorrectales/química , Proteína de la Poliposis Adenomatosa del Colon/genética , Adulto , Anciano , Anciano de 80 o más Años , Cadherinas/genética , Neoplasias Colorrectales/genética , Epitelio/química , Femenino , Expresión Génica , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana EdadRESUMEN
Deletions involving chromosome 6q have been reported in a number of human cancers such as ovarian and breast tumours as well as haematopoietic malignancies. It seems that this region might contain tumour-suppressor genes. Putative natural killer cell lymphomas/leukaemias (NKLL) represent a group of recently characterized haematolymphoid malignancies sharing an immunophenotype of CD3/Leu4- CD3epsilon+ CD56+, a genotype of germline T-cell receptor genes, and have a close association with Epstein-Barr virus (EBV). Deletion at 6q21-q25 was demonstrated in three recently reported cases of NKLL. Here we investigated the possible involvement of 6q deletions in the pathogenesis, and especially the tumorigenesis of NKLL. The regions of D6S1574 (6p25), DS276 (6p12), D6S257 (6q11), D6S434 (6q14), D6S287 (6q15), D6S292 (6q21), D6S308 (6q22), D6S264 (6q25), and D6S446 (6q26) were analysed by PCR in 25 cases of NKLL, including seven cases with chronic NK leukaemia, six with acute NK leukaemia and 12 with NK lymphoma. 6q deletions, especially 6q15-25, were frequently detected, but 6p deletions were not detected in any cases. Analysis of 6q21 showed possible deletion in two of seven cases (29%) with chronic NK leukaemia, three of six (50%) with acute leukaemia, and 12 of 12 (100%) with NK lymphoma. The frequency of deletion increased in clinical phases. In three cases with lymphoma, fluorescence in situ hybridisation was performed, which confirmed 6q21 deletion in two cases, although 6q telomeric and centromeric regions were preserved. The other case failed to show deletion. Our results suggest that 6q deletion, especially 6q21-25, might be involved in NKLL tumorigenesis, and support the presence of the tumour suppressor genes associated with the development of NKLL.
Asunto(s)
Deleción Cromosómica , Cromosomas Humanos Par 6/genética , Células Asesinas Naturales/patología , Leucemia de Células T/genética , Linfoma de Células T/genética , Adolescente , Adulto , Anciano , Niño , Preescolar , Mapeo Cromosómico , Infecciones por Virus de Epstein-Barr/complicaciones , Femenino , Humanos , Hibridación Fluorescente in Situ , Células Asesinas Naturales/virología , Leucemia de Células T/etiología , Leucemia de Células T/virología , Pérdida de Heterocigocidad , Linfoma de Células T/etiología , Linfoma de Células T/virología , Masculino , Persona de Mediana EdadRESUMEN
Tumor multicentricity is occasionally observed in esophageal squamous cell carcinoma (SCC). We studied five surgically resected superficial multifocal esophageal SCCs for p53 gene mutation and genetic instability, using DNA extracted from microdissected areas. A total of 38 target areas (TAs) were analyzed in SCC, dysplasia, basal cell hyperplasia (BCH) and normal squamous epithelium. Analysis of the replication error (RER) at 10 microsatellite loci showed microsatellite instability in all TAs, as well as in normal squamous epithelium. p53 gene mutation was identified in 28.9% (11/38 TAs). All cases showed a common missense mutation in exon 8 at codon 273 (CGT-->CAT, Arg-->His), which was DNA contact mutation in the S10 beta strand. In association with microsatellite alterations, 7 of 9 TAs with p53 mutation in exon 8 at codon 273 also showed loss of heterozygosity (LOH) of p53 gene. LOH of p53 gene was detected in 83.8% (31/37 TAs). LOH at D2S123 on 2p16 near MSH2 gene and at D3S1611 on 3p22 near MLH1 gene was detected in 65.4% (17/26) and 71.4% (10/14) TAs, respectively. Frequencies of LOH at p53 and D2S123 were similar in non-cancerous areas and SCCs. LOH of p53 and D2S123 were found in 50% (5/10 TAs) of non-cancerous areas and 60% (9/15 TAs) of SCCs. Our results suggest that genetic instability induces esophageal tumor multicentricity, and that p53 gene contact mutation together with LOH are early events of the multistage carcinogenesis of multifocal primary esophageal SCC.