RESUMEN
BACKGROUND: Peritoneal metastasis is a frequent cause of death in patients with gastric cancer. The aim of this study was to identify molecules responsible for mediating peritoneal metastasis of gastric cancer. METHODS: Transcriptome and bioinformatics analyses were conducted to identify molecules associated with peritoneal metastasis. The therapeutic effects of intraperitoneally administered small interfering (si) RNA were evaluated using mouse xenograft models. Expression of mRNA and protein was determined in gastric tissues from patients with gastric cancer. RESULTS: Synaptotagmin XIII (SYT13) was expressed at significantly higher levels in patients with peritoneal recurrence, but not in those with hepatic or distant lymph node recurrence. Inhibition of SYT13 expression in a gastric cancer cell line transfected with SYT13-specific siRNA (siSYT13) was associated with decreased invasion and migration ability of the cells, but not with proliferation and apoptosis. Intraperitoneal administration of siSYT13 significantly inhibited the growth of peritoneal nodules and prolonged survival in mice. In an analysis of 200 patients with gastric cancer, SYT13 expression in primary gastric cancer tissues was significantly greater in patients with peritoneal recurrence or metastasis. A high level of SYT13 expression in primary gastric cancer tissues was an independent risk factor for peritoneal recurrence. CONCLUSION: SYT13 expression in gastric cancer is associated with perioneal metatases and is a potential target for treatment.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias Peritoneales/secundario , Neoplasias Gástricas/patología , Sinaptotagminas/metabolismo , Anciano , Animales , Biomarcadores de Tumor/antagonistas & inhibidores , Línea Celular Tumoral , Biología Computacional , Femenino , Estudios de Seguimiento , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad , Trasplante de Neoplasias , Neoplasias Peritoneales/metabolismo , Neoplasias Peritoneales/prevención & control , Interferencia de ARN , ARN Interferente Pequeño/uso terapéutico , Tratamiento con ARN de Interferencia , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/terapia , Sinaptotagminas/antagonistas & inhibidores , TranscriptomaRESUMEN
OBJECTIVE: Joint instability induced by anterior cruciate ligament (ACL) transection is commonly considered as a predisposing factor for osteoarthritis (OA) of the knee; however, the influence of re-stabilization on the protection of articular cartilage is unclear. The aim of this study was to evaluate the effect of joint re-stabilization on articular cartilage using an instability and re-stabilization ACL transection model. DESIGN: To induce different models of joint instability, our laboratory created a controlled abnormal joint movement (CAJM) group and an anterior cruciate ligament transection group (ACL-T). Seventy-five Wistar male rats were randomly assigned to the CAJM (n = 30), ACL-T (n = 30), or no treatment (INTACT) group (n = 15). Cartilage changes were assessed with soft X-ray analysis, histological and immunohistochemistry analysis, and real-time polymerase chain reaction (PCR) analysis at 2, 4, and 12 weeks. RESULTS: Joint instability, as indicated by the difference in anterior displacement between the CAJM and ACL-T groups (P < 0.001), and cartilage degeneration, as evaluated according to the Osteoarthritis Research Society International (OARSI) score, were significantly higher in the ACL-T group than the CAJM group at 12 weeks (P < 0.001). Moreover, joint re-stabilization maintained cartilage structure (thickness [P < 0.001], surface roughness [P < 0.001], and glycosaminoglycan stainability [P < 0.001]) and suppressed tumor necrosis factor-alpha (TNF-α) and caspase-3 at 4 weeks after surgery. CONCLUSION: Re-stabilization of joint instability may suppress inflammatory cytokines, thereby delaying the progression of OA. Joint instability is a substantial contributor to cartilage degeneration.
Asunto(s)
Cartílago Articular/patología , Inestabilidad de la Articulación/prevención & control , Animales , Ligamento Cruzado Anterior/patología , Modelos Animales de Enfermedad , Inestabilidad de la Articulación/complicaciones , Masculino , Osteoartritis de la Rodilla/etiología , Osteoartritis de la Rodilla/prevención & control , Ratas , Ratas WistarRESUMEN
Despite improvements in surgical techniques, perioperative management, and multidisciplinary therapy, treatment outcomes of patients with esophageal squamous cell carcinoma (ESCC) remain poor. Therefore, development of novel molecular biomarkers, which either predict patient survival or become therapeutic targets, is urgently required. In the present study, to facilitate early detection of ESCC and predict its clinical course, we investigated the relationship of the serum level of melanoma-associated antigen (MAGE)-D4 to patients' clinicopathological characteristics. Using quantitative real-time reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assays, we determined the levels of MAGE-D4 mRNA and protein in cell lysates and conditioned medium of cultures, respectively, of nine ESCC cell lines. Further, we determined MAGE-D4 levels in serum samples collected from 44 patients with ESCC who underwent radical esophagectomy without neoadjuvant therapy as well as from 40 healthy volunteers. Samples of conditioned medium and cell lysates contained comparable levels of MAGE-D4 that correlated closely with the levels of MAGE-D4 mRNA. Preoperative MAGE-D4 levels in the sera of 44 patients with ESCC, which varied from 0 to 2,354 pg/mL (314 ± 505 pg/mL, mean ± standard deviation), were significantly higher compared with those of healthy volunteers. By setting the cutoff at the highest value for healthy volunteers (50 pg/mL), the MAGE-D4-positive group of patients was more likely to have shorter disease-specific and disease-free survival compared with those of the MAGE-D4-negative group, although the differences were not statistically significant. Our results indicate that the elevation of preoperative serum MAGE-D4 levels in some patients with ESCC was possibly caused by excess production of MAGE-D4 by tumor cells followed by its release into the circulation. Clinical implications of serum MAGE-D4 levels should be validated in a large population of patients with ESCC.
Asunto(s)
Antígenos de Neoplasias/sangre , Biomarcadores de Tumor/sangre , Carcinoma de Células Escamosas/sangre , Neoplasias Esofágicas/sangre , Adulto , Anciano , Anciano de 80 o más Años , Antígenos de Neoplasias/genética , Biomarcadores de Tumor/genética , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/cirugía , Estudios de Casos y Controles , Línea Celular Tumoral , Supervivencia sin Enfermedad , Ensayo de Inmunoadsorción Enzimática , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/cirugía , Carcinoma de Células Escamosas de Esófago , Esofagectomía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , ARN Mensajero/sangre , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
To pursue an urgently needed treatment target for esophageal cancer (EC), we investigated the function of the recently discovered melanoma-associated antigen (MAGE)-D4 in squamous cell EC. MAGE-D4 messenger RNA (mRNA) expression was analyzed in nine EC cell lines using quantitative reverse transcription polymerase chain reaction. In 65 surgical specimens of squamous cell EC with no prior neoadjuvant therapy, MAGE-D4â mRNA expression in EC tissues and corresponding normal tissues was analyzed and compared, and evaluated in terms of clinicopathological factors. In representative cases, MAGE-D4 protein distribution was analyzed immunohistochemically. The heterogeneity of MAGE-D4â mRNA expression was confirmed in EC cell lines by quantitative reverse transcription polymerase chain reaction. In surgical specimens, MAGE-D4â mRNA expression was significantly higher in EC tissues than in corresponding normal tissues (P < 0.001). Patients with the highest MAGE-D4â mRNA expression in EC tissues (top quartile, n = 17) had significantly shorter overall survival than patients with low expression (2-year survival: 44% and 73%, respectively, P = 0.006). Univariate analysis identified age (≥65 years), lymphatic involvement, and high MAGE-D4â mRNA expression as significant prognostic factors; high MAGE-D4â mRNA expression was also an independent prognostic factor in multivariable analysis (hazard ratio: 2.194; P = 0.039) and was significantly associated with Brinkman index (P = 0.008) and preoperative carcinoembryonic antigen level (P = 0.002). Immunohistochemical MAGE-D4b expression was consistent with MAGE-D4â mRNA profiling. Our results suggest that MAGE-D4 overexpression influences tumor progression, and MADE-D4 can be a prognostic marker and a potential molecular target in squamous cell EC.
Asunto(s)
Antígenos de Neoplasias/metabolismo , Biomarcadores de Tumor/metabolismo , Carcinoma de Células Escamosas/metabolismo , Neoplasias Esofágicas/metabolismo , ARN Mensajero/metabolismo , Anciano , Anciano de 80 o más Años , Antígenos de Neoplasias/genética , Biomarcadores de Tumor/genética , Carcinoma de Células Escamosas/genética , Línea Celular Tumoral , Progresión de la Enfermedad , Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas de Esófago , Femenino , Humanos , Masculino , Persona de Mediana Edad , PronósticoRESUMEN
This case report describes an exceptional case of ectopic adrenal cortex tissue (EACT) in the spermatic cord associated with ipsilateral cryptorchidism in an adult. While both EACT and cryptorchidism are fairly common congenital anomalies in boys, adult cases are uncommon. Although the spermatic cord is a known site of EACT, the reports of its association with cryptorchidism have been limited to child cases. During surgery, undescended testis was discovered and incidentally ectopic adrenal tissue along the spermatic cord was also identified. This combination of developmental aberrations in the adult has not been described, and the clinicopathological findings are reported with a brief literature review.
Asunto(s)
Corteza Suprarrenal , Coristoma/complicaciones , Criptorquidismo/complicaciones , Cordón Espermático/patología , Coristoma/patología , Coristoma/cirugía , Criptorquidismo/patología , Criptorquidismo/cirugía , Enfermedades de los Genitales Masculinos/complicaciones , Enfermedades de los Genitales Masculinos/patología , Humanos , Masculino , Persona de Mediana Edad , Cordón Espermático/cirugía , Resultado del Tratamiento , Procedimientos Quirúrgicos Urológicos MasculinosRESUMEN
Capecitabine is a fluoropyrimidine-based drug that offers physicians a more convenient treatment for advanced colorectal cancer (CRC), with manageable toxicity and antitumor activity comparable to that of continuous-infusion therapies with 5-fluorouracil (5-FU). However, there are no validated and established predictive factors for clinical outcome of capecitabine efficacy in CRC. The gene expressions of the pyrimidine metabolism enzymes dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP) and thymidylate synthase (TS) have previously been shown to be response determinants of fluoropyrimidine-based drugs in various tumors. Therefore, we investigated whether intratumoral mRNA expression levels of these genes are also associated with the clinical outcome of patients with metastatic CRC treated with first-line capecitabine. Thirty-seven patients with metastatic CRC were enrolled in this study and treated with single agent capecitabine. The intratumoral mRNA levels of DPD, TP and TS were assessed from paraffin-embedded tissue samples using laser-capture-microdissection methods and quantitative real-time PCR. There were 20 women and 17 men with a median age of 61 years (range 49-74). The median progression-free survival was 6.7 months (95% CI, 4.8-11.6 months), with a median follow-up of 14.4 months (range 1.3-18.7 months). Complete response was observed in 1 (3%), partial response in 6 (20%), stable disease in 14 (47%) and progressive disease in 9 (30%) patients (response was inevaluable in 7 patients). Higher gene expression levels of DPD were associated with resistance to capecitabine (P=0.032; Kruskal-Wallis test). Patients with a lower mRNA amount of DPD (Asunto(s)
Neoplasias Colorrectales/tratamiento farmacológico
, Desoxicitidina/análogos & derivados
, Dihidrouracilo Deshidrogenasa (NADP)/metabolismo
, Fluorouracilo/análogos & derivados
, Regulación Neoplásica de la Expresión Génica
, Anciano
, Antineoplásicos/farmacología
, Capecitabina
, Desoxicitidina/farmacología
, Progresión de la Enfermedad
, Supervivencia sin Enfermedad
, Femenino
, Fluorouracilo/farmacología
, Humanos
, Masculino
, Persona de Mediana Edad
, Metástasis de la Neoplasia
, Proyectos Piloto
, ARN Mensajero/metabolismo
, Resultado del Tratamiento
RESUMEN
Thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD) and thymidine phosphorylase (TP) are predictive markers for tumor response to 5-fluorouracil-based therapies. To determine whether gene expression values measured in primary cancer tissue would be useful for prediction of response of lymph node metastases, the expressions of these genes were quantitatively analyzed in 35 pairs of primary colorectal cancer (CRC) and corresponding lymph node metastases using real-time PCR. DPD and TP mRNA levels were significantly lower in the primary colorectal tumor and lymph node metastases compared with the normal adjacent stroma tissue (p<0.01), whereas TS mRNA levels were significantly higher in the primary tumor and lymph node metastases than in the normal adjacent tissue (p<0.001). Median gene expression levels of TP and TS did not differ significantly between primary colorectal tumor and corresponding lymph node metastasis but median DPD gene expression levels in the lymph node metastases were significantly higher compared to matched primary colorectal tumors (p=0.015). There was a significant correlation for DPD, TP and TS gene expression levels between primary colorectal tumor specimens and the matched lymph node metastasis. These results suggest that biopsies of the tumor of origin may be valid for determining predictive markers for chemotherapy response in patients with metastatic CRC.
Asunto(s)
Antimetabolitos Antineoplásicos/metabolismo , Neoplasias Colorrectales/metabolismo , Fluorouracilo/metabolismo , Regulación Neoplásica de la Expresión Génica , Anciano , Anciano de 80 o más Años , Antimetabolitos Antineoplásicos/uso terapéutico , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/genética , Dihidrouracilo Deshidrogenasa (NADP)/genética , Dihidrouracilo Deshidrogenasa (NADP)/metabolismo , Femenino , Fluorouracilo/uso terapéutico , Humanos , Ganglios Linfáticos/metabolismo , Metástasis Linfática , Masculino , Persona de Mediana Edad , ARN Mensajero/metabolismo , Timidina Fosforilasa/genética , Timidina Fosforilasa/metabolismo , Timidilato Sintasa/genética , Timidilato Sintasa/metabolismoRESUMEN
BACKGROUND: The objective of this study was to examine the clinical outcomes of immediate breast reconstruction using perforator flaps from different donor sites, and to characterize the trends among these flaps. METHODS: We retrospectively reviewed 136 consecutive patients who underwent immediate breast reconstruction using free flaps after skin-sparing mastectomy (SSM) or nipple-sparing mastectomy (NSM). The whole breast was pathologically analyzed in 5-mm sections. Breast reconstruction was performed using the deep inferior epigastric perforator (DIEP) flap, gluteal artery perforator (GAP) flap, and posterior medial thigh perforator (PMTP) flap. Patient characteristics were compared among donor sites. RESULTS: NSM was converted to SSM because of intraoperative subareolar tumor positivity in 7 of 107 patients. Eleven patients had positive margins in permanent sections. All but one patient had a positive horizontal margin in the peripheral direction. The 5-year recurrence-free survival rate was 91.9%. The locoregional recurrence rate was 5.1% with a mean follow-up observation period of 75 months. DEIP, GAP, and PMTP flaps were used in 64 (47.1%), 38 (27.9%), and 34 (25.0%) patients, retrospectively. DIEP flaps were used in older patients and those with a higher body mass index. GAP flaps were used in younger patients. DIEP and GAP flaps were used for larger breasts, and PMTP flaps for smaller breasts. CONCLUSION: NSM or SSM with immediate perforator flap breast reconstruction is an oncologically acceptable surgical option. We believe that age, desire to have children, body mass index, and excised breast volume are valuable factors for selecting the optimal donor site.
Asunto(s)
Neoplasias de la Mama/cirugía , Mamoplastia/métodos , Mastectomía Subcutánea/métodos , Colgajo Perforante/trasplante , Sitio Donante de Trasplante/cirugía , Centros Médicos Académicos , Adulto , Análisis de Varianza , Neoplasias de la Mama/patología , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Humanos , Japón , Persona de Mediana Edad , Recurrencia Local de Neoplasia/patología , Recurrencia Local de Neoplasia/fisiopatología , Satisfacción del Paciente/estadística & datos numéricos , Colgajo Perforante/irrigación sanguínea , Cuidados Posoperatorios/métodos , Estudios Retrospectivos , Medición de Riesgo , Estadísticas no Paramétricas , Factores de Tiempo , Trasplante Autólogo , Cicatrización de Heridas/fisiología , Adulto JovenRESUMEN
BACKGROUND: Although sentinel lymph node biopsy (SLNB) is a standard staging method for assessing nodal status of breast cancer patients, SLNB after neoadjuvant chemotherapy (NAC) remains controversial. The aim of this study was to validate the practicality and accuracy of SLNB by our modified Indigocarmine blue dye methods following NAC. METHODS: One hundred consecutive cases with breast cancers treated by NAC were enrolled in this study. After NAC, all patients underwent SLNB performed by our modified Indigocarmine blue dye methods without radioisotope, followed by back-up axillary lymph node dissection (ALND). RESULTS: Sentinel nodes (SNs) were identified in 94 cases (identification rate, 94%); the accuracy was 94.7% (89/94 cases); and the false negative rate (FNR) 13.5% (5/37 cases). For cases with vs. without clinically evident metastatic nodes before NAC, the identification rate was 92.4% (61/66 cases) vs. 97.1% (33/34 cases); the accuracy 91.8% (56/61 cases) vs. 97.0% (32/33 cases) and the FNR 16.1% (5/31 cases) vs. 0% (0/6 case), respectively. There were six patients without identified SNs, three of them had metastatic nodes. False negatives occurred in five cases; in four, fewer than two sentinel nodes had been removed. CONCLUSION: Following NAC, the accuracy of SLNB by modified Indigocarmine blue dye methods is adequate compared with other tracers. In patients in whom no SNs have been identified, lymphatic metastasis is likely and therefore ALND is recommended. For patients with cN0 prior to NAC, SLNB by modified Indigocarmine blue dye methods is clinically feasible, though controversial for patients with positive nodes.
Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Escisión del Ganglio Linfático , Ganglios Linfáticos/patología , Biopsia del Ganglio Linfático Centinela/métodos , Adulto , Anciano , Antineoplásicos , Axila , Quimioterapia Adyuvante , Colorantes , Reacciones Falso Negativas , Estudios de Factibilidad , Humanos , Carmin de Índigo , Ganglios Linfáticos/cirugía , Metástasis Linfática , Persona de Mediana Edad , Terapia Neoadyuvante , Estadificación de Neoplasias , Estudios ProspectivosRESUMEN
BACKGROUND: The impact of neoadjuvant chemotherapy (NACT) on immediate free flap breast reconstruction remains controversial. Furthermore, the oncological outcomes of immediate free flap breast reconstruction after skin-sparing mastectomy (SSM) following NACT remain unclear. This study aimed to investigate the surgical complications and oncological outcomes of immediate perforator flap reconstruction after SSM following NACT. METHODS: A total of 201 consecutive patients with indications for immediate perforator flap reconstruction after SSM were included between 2004 and 2012. Surgical and oncological outcomes were compared between patients with and without NACT. RESULTS: There were 38 patients in the NACT group and 163 in the non-NACT control group. The median age of the NACT group was 39.5 years, which was significantly younger than the control group (43.0 years; P < 0.05). Patients in the NACT group also had more advanced and aggressive disease (P < 0.05). There was no significant difference in the frequency of surgical complications between the groups, no difference in the type of complications, and no significant difference in the frequencies of major and minor complications. No patients in the NACT group had delayed adjuvant therapy. Eight patients (4%) developed recurrences, with a median follow-up time of 3.0 years. Local recurrences occurred in three control patients but no patients in the NACT group. CONCLUSION: NACT does not affect short-term or interim outcomes after immediate perforator flap reconstruction and may thus represent a safe and practical treatment option for the multidisciplinary treatment of breast cancer.
Asunto(s)
Neoplasias de la Mama/cirugía , Carcinoma Ductal de Mama/cirugía , Carcinoma Intraductal no Infiltrante/cirugía , Carcinoma Lobular/cirugía , Mamoplastia/métodos , Mastectomía Subcutánea/métodos , Terapia Neoadyuvante , Colgajo Perforante , Adulto , Anticuerpos Monoclonales Humanizados/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Carboplatino/uso terapéutico , Carcinoma Ductal de Mama/tratamiento farmacológico , Quimioterapia Adyuvante , Estudios de Cohortes , Ciclofosfamida/uso terapéutico , Docetaxel , Epirrubicina/uso terapéutico , Etopósido/uso terapéutico , Femenino , Fluorouracilo/uso terapéutico , Humanos , Persona de Mediana Edad , Paclitaxel/administración & dosificación , Estudios Retrospectivos , Taxoides/administración & dosificación , Trastuzumab , Resultado del Tratamiento , Adulto JovenRESUMEN
Mutagenesis studies were carried out to examine the effects of replacement of either the nucleophile Glu-236 or the acid/base Glu-128 residue of the F/10 xylanase by a His residue. To our surprise, the affinity for the p-nitrophenyl-beta-D-xylobioside substrate was increased by 10(3)-fold in the case of the mutant E128H enzyme compared with that of the wild-type F/10 xylanase. The catalytic activity of the mutant enzymes was low, despite the fact that the distance between the nucleophilic atom (an oxygen in the native xylanase and a nitrogen in the mutant) and the alpha-carbon was barely changed. Thus, the alteration of the acid/base functionality (Glu-128 to His mutation) provided a significantly favorable interaction within the E128H enzyme/substrate complex in the ground state, accompanying a reduction in the stabilization effect in the transition state.
Asunto(s)
Disacáridos/metabolismo , Streptomyces/enzimología , Xilosidasas/metabolismo , Endo-1,4-beta Xilanasas , Ácido Glutámico/genética , Ácido Glutámico/metabolismo , Histidina/genética , Histidina/metabolismo , Cinética , Mutagénesis Sitio-Dirigida , Streptomyces/genética , Xilosidasas/genéticaRESUMEN
Although the amino acid homology in the catalytic domain of FXYN xylanase from Streptomyces olivaceoviridis E-86 and Cex xylanase from Cellulomonas fimi is only 50%, an active chimeric enzyme was obtained by replacing module 10 in FXYN with module 10 from Cex. In the family F/10 xylanases, module 10 is an important region as it includes an acid/base catalyst and a substrate binding residue. In FXYN, module 10 consists of 15 amino acid residues, while in Cex it consists of 14 amino acid residues. The Km and kcat values of the chimeric xylanase FCF-C10 for PNP-xylobioside (PNP-X2) were 10-fold less than those for FXYN. CD spectral data indicated that the structure of the chimeric enzyme was similar to that of FXYN. Based on the comparison of the amino acid sequences of FXYN and Cex in module 10, we constructed four mutants of FXYN. When D133 or S135 of FXYN was deleted, the kinetic properties were not changed from those of FXYN. By deletion of both D133 and S135, the Km value for PNP-X2 decreased from the 2.0 mM of FXYN to 0.6 mM and the kcat value decreased from the 20 s(-1) of FXYN to 8.7 s(-1). Insertion of Q140 into the doubly deleted mutant further reduced the Km value to 0.3 mM and the kcat value to 3.8 s(-1). These values are close to those for the chimeric enzyme FCF-C10. These results indicate that module 10 itself is able to accommodate changes in the sequence position of amino acids which are critical for enzyme function. Since changes of the spatial position of these amino acids would be expected to result in enzyme inactivation, module 10 must have some flexibility in its tertiary structure. The structure of module 10 itself also affects the substrate specificity of the enzyme.
Asunto(s)
Endo-1,4-beta Xilanasas , Streptomyces/enzimología , Xilosidasas/genética , beta-Glucosidasa/genética , Secuencia de Aminoácidos , Sitios de Unión , Dicroismo Circular , Glicósidos/metabolismo , Enlace de Hidrógeno , Cinética , Modelos Moleculares , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Mutación , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Proteínas Recombinantes de Fusión , Alineación de Secuencia , Especificidad por Sustrato , Xilano Endo-1,3-beta-Xilosidasa , Xilosidasas/química , beta-Glucosidasa/químicaRESUMEN
Endogenous lipid droplets were prepared by subjecting fat cells to hypotonic shock and to Triton X-100 treatment. The structure of the endogenous lipid droplet fraction was examined by scanning and transmission electron microscopies. Neither intact fat cells nor disrupted cell membranes were detectable in the endogenous lipid droplet fraction. With this endogenous substrate, epinephrine elicited lipolysis with either hormone-sensitive lipase or lipoprotein lipase, but no cyclic AMP-protein kinase mediated stimulation of lipolysis was observed. On the other hand, epinephrine did not stimulate lipolysis when triolein emulsified with arabic gum was used as substrate. With the latter exogenous substrate, however, cyclic AMP-protein kinase was found to stimulate lipolysis with hormone-sensitive lipase as enzyme. These results agree with the proposal of Wise and Jungas that the epinephrine-stimulated increase of hydrolysis of endogenous fat is not mediated through cyclic AMP-protein kinase. A possible mechanism of hydrolysis of endogenous fat by induction of lipolysis by epinephrine in fat cells is discussed.
Asunto(s)
Tejido Adiposo/metabolismo , Epinefrina/farmacología , Lipólisis/efectos de los fármacos , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/ultraestructura , Animales , Metabolismo de los Lípidos , Masculino , Microscopía Electrónica , Microscopía Electrónica de Rastreo , Ratas , Ratas EndogámicasRESUMEN
The aim of this study was to compare the morphological development of dorsal root ganglion neurons in embryonic and early postnatal chicks and rats. The three-dimensional architecture of neurons was observed in ganglia in situ and in dissociated neurons by scanning electron microscopy after removal of the capsule and connective tissue. The percentages of neurons at different developmental stages were determined. The general morphological changes in the chick resembled those in the rat but the timing was different. In both chick and rat, the majority of neurons were bipolar at early stages of development (embryonic day 6 in chick and day 14 in rats) and later underwent pseudo-unipolarization to become mostly unipolar neurons at hatching or birth. This maturation event started at an earlier stage in chick embryos than in rats, with 57% unipolar neurons in chick and only 7% in rat on embryonic day 14. However, just after hatching or birth, at day 22 of development, a larger proportion of immature unipolar neurons remained in chicks (13%) than in rats (3%). We conclude that these differences should be taken into consideration in designing experiments on dorsal root ganglion neurons grown in tissue culture.
Asunto(s)
Animales Recién Nacidos/crecimiento & desarrollo , Embrión de Pollo/embriología , Ganglios Espinales/embriología , Neuronas Motoras/ultraestructura , Ratas/embriología , Animales , Recuento de Células , Femenino , Ganglios Espinales/ultraestructura , Microscopía Electrónica de Rastreo , Embarazo , Especificidad de la EspecieRESUMEN
The effect of two cationic drugs (chlorpromazine and isoxsuprine) on the suspension viscosity of human erythrocytes were examined, comparing with the effect of anionic drugs. (1) As increasing the drug concentrations, the cationic drugs transformed the erythrocytes to stomatocytes, then to spherostomatocytes, while trinitrobenzene sulfonate, dehydroepiandrosterone sulfate and lysolecithin induced echinocytes, as well known. (2) The suspension viscosity decreased in parallel with the appearance of spherostomatocytes, but it increased in echinocytosis. (3) The membrane fluidity, measured by spin label method, was not a major determinant for the suspension viscosity in these cases, because of no systematic correlation. (4) The rheoscopic observation under shear force demonstrated that the spherostomatocytes deformed easily to ellipsoid with smooth cell surface, while the echinocytes less easily deformed to ellipsoid on which the small spikes persisted at higher shear. These distinct difference in deformed shape under high shear force could be related to the decreased suspension viscosity of spherostomatocytes. (5) In addition, the transformation to spherostomatocytes, thus the decreased viscosity, was primarily determined by the intramembraneous drug concentration.
Asunto(s)
Clorpromazina/farmacología , Eritrocitos/efectos de los fármacos , Isoxsuprina/farmacología , Viscosidad Sanguínea , Eritrocitos/fisiología , Humanos , Suspensiones , ViscosidadRESUMEN
INTRODUCTION: We report herein a new method of transumbilical laparoscopic surgery using a GelPort through an umbilical zigzag skin incision. The method involves collaborating with plastic surgeons to ensure the procedure was minimally invasive. MATERIALS AND SURGICAL TECHNIQUE: After marking a zigzag skin incision in the umbilical region, the skin was incised along this line. Then, a GelPort double-ring wound retractor was inserted through the incision, which enlarged the diameter of the fascial opening to 6 cm. The Gelport was latched on the wound retractor ring, following the inflation of the pneumoperitoneum by CO (2). One or more additional ports were inserted as necessary. All operations were performed in the standard fashion. The specimen was easily extracted from the abdomen through the umbilical incision, and anastomosis was performed. Using the above method, we performed the following procedures: one total gastrectomy, one distal gastrectomy, three gastric local resections, five right hemicolectomies, two high anterior resections, three cholecystectomies, and seven transabdominal preperitoneal hernioplasties. All cases were accomplished without any complications using this method. The wounds of the umbilical region were almost "scarless" in all cases. DISCUSSION: We developed an umbilical zigzag skin incision technique to perform abdominal laparoscopic operations using a GelPort, with a minimal number of skin incisions. We consider that our method reduces the technical difficulties associated with laparoscopic surgery and maintains cosmesis.
Asunto(s)
Colectomía/métodos , Gastrectomía/métodos , Herniorrafia/métodos , Laparoscopía/métodos , Ombligo/cirugía , Colecistectomía Laparoscópica/instrumentación , Colecistectomía Laparoscópica/métodos , Colectomía/instrumentación , Gastrectomía/instrumentación , Herniorrafia/instrumentación , Humanos , Laparoscopía/instrumentaciónRESUMEN
A left brachial plexus and axillary artery of bonobo (Pan paniscus) were examined, and the interrelation between the brachial plexus and the axillary artery was discussed. This is the first report of the brachial plexus and the axillary artery of bonobo. The bonobo brachial plexus formed very similar pattern to that of other ape species and human. On the other hand, the branches of the bonobo axillary artery had uncommon architecture in comparison with human case. The axillary artery did not penetrate the brachial plexus and passes through all way along anterior to the brachial plexus. Only 4.9% of human forelimbs have this pattern. Moreover, the brachial artery runs through superficially anterior to branches of the brachial plexus.
Asunto(s)
Arteria Axilar/anatomía & histología , Plexo Braquial/anatomía & histología , Pan paniscus/anatomía & histología , Animales , Arteria Braquial/anatomía & histología , Plexo Braquial/irrigación sanguínea , Miembro Anterior/irrigación sanguínea , Procesamiento de Imagen Asistido por Computador , MasculinoRESUMEN
BACKGROUND: The use of radioisotopes (RIs) is regulated and not all institutions have nuclear medicine facilities for sentinel node biopsy (SNB). We previously reported blue dye-assisted four-node axillary sampling (4NAS/dye) to be a suitable method for detecting sentinel nodes (SNs) without RIs. Here, we present an interim report on an observational study of this technique. METHODS: From May 2003 to June 2008, 234 early breast cancer patients underwent SNB with 4NAS/dye. Lymphatic mapping was performed by injection of patent blue, and axillary sampling was performed until 4 SNs were detected. Patients with metastatic SNs underwent axillary lymph node dissection (ALND) at levels I and II, while SN-negative patients did not undergo further axillary procedures. RESULTS: The SN identification rate was 99%. In total, 44 patients were diagnosed with metastatic disease by using the 4NAS/dye technique and underwent ALND; the remaining 189 patients did not undergo ALND (the SNB group). After a median follow-up period of 54 months, only 1 patient (0.5%) in the SNB group developed axillary recurrence. For the 4NAS/dye procedure, blue SNs were harvested in 220 patients (94%) and only unstained SNs were harvested in 13 patients (6%). Among the 44 patients with SN metastases, foci were found in blue SNs in 37 patients (84%), while they were found in only unstained SNs in 7 patients (16%). CONCLUSIONS: SNB with 4NAS/dye is a safe and reliable technique for treatment of early breast cancer patients. This technique may be particularly useful for surgeons who do not have access to radioisotope facilities.