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Mixed endometrial stromal and smooth muscle tumor of the uterus is a rare occurrence, and it is truly challenging to diagnose or dif- ferentiate mesenchymal tumors of the uterine corpus, due to their many overlapping features. In most cases, the gross pathology of mixed endometrial stromal and smooth muscle tumor differs from that of pure endometrial stromal and pure smooth muscle tumors. A 59-year-old postmenopausal woman presented with vaginal spotting, low abdominal pain, and an uterine mass. Subsequent pelvic magnetic resonance imaging revealed a 4.0x3.8x3.4-cm sized uterine mass with enhancement. The mass showed restricted diffusion on diffusion-weighted images, and thus, was suspected to be uterine sarcoma rather than degenerative leiomyoma. Levels of tumor markers, CA 125, CA 19-9, and SCC, were within their normal ranges. The patient underwent exploratory laparotomy. Morphological and immunohistochemical evaluations were performed, and a final diagnosis of mixed endometrial stromal and smooth muscle tumor of the uterus was rendered. Her postoperative course was uneventful, and aromatase inhibitor adjuvant therapy was administered.
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Neoplasias Endometriales/patología , Tumores Estromáticos Endometriales/patología , Neoplasias Complejas y Mixtas/patología , Tumor de Músculo Liso/patología , Desmina/metabolismo , Neoplasias Endometriales/metabolismo , Tumores Estromáticos Endometriales/metabolismo , Femenino , Humanos , Persona de Mediana Edad , Neoplasias Complejas y Mixtas/metabolismo , Neprilisina/metabolismo , Tumor de Músculo Liso/metabolismoRESUMEN
OBJECTIVE: Cartilage regenerative procedures using the cell-based tissue engineering approach involving mesenchymal stem cells (MSCs) have been receiving increased interest because of their potential for altering the progression of osteoarthritis (OA) by repairing cartilage lesions. The aim of this study was to investigate the clinical and magnetic resonance imaging (MRI) outcomes of MSC implantation in OA knees and to determine the association between clinical and MRI outcomes. DESIGN: Twenty patients (24 knees) who underwent arthroscopic MSC implantation for cartilage lesions in their OA knees were evaluated at 2 years after surgery. Clinical outcomes were evaluated according to the International Knee Documentation Committee (IKDC) score and the Tegner activity scale, and cartilage repair was assessed according to the MRI Osteoarthritis Knee Score (MOAKS) and Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) score. RESULTS: The clinical outcomes significantly improved (P < 0.001 for both). The cartilage lesion grades (as described in MOAKS [grades for size of cartilage-loss area and percentage of full-thickness cartilage loss]) at follow-up MRI were significantly better than the preoperative values (P < 0.001 for both). The clinical outcomes at final follow-up were significantly correlated with the MOAKS and MOCART score at follow-up MRI (P < 0.05 for all). CONCLUSIONS: Considering the encouraging clinical and MRI outcomes obtained and the significant correlations noted between the clinical and MRI outcomes, MSC implantation seems to be useful for repairing cartilage lesions in OA knees. However, a larger sample size and long-term studies are needed to confirm our findings.
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Cartílago Articular/patología , Trasplante de Células Madre Mesenquimatosas/métodos , Osteoartritis de la Rodilla/terapia , Anciano , Artroscopía , Estudios de Cohortes , Femenino , Adhesivo de Tejido de Fibrina/uso terapéutico , Humanos , Escala de Puntuación de Rodilla de Lysholm , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/patología , Osteoartritis de la Rodilla/fisiopatología , Satisfacción del Paciente , Estudios Prospectivos , Índice de Severidad de la Enfermedad , Adhesivos Tisulares/uso terapéutico , Resultado del TratamientoRESUMEN
The aim of this study was to evaluate the reliability of using tumour grade and cell type on preoperative endometrial biopsy for the selection of patients for conservative hormone treatment. We retrospectively reviewed results of 643 patients with endometrial carcinoma for tumour grade and 817 for tumour cell type who underwent endometrial biopsy followed by surgery. Of the 357 patients with a grade 1 tumour on preoperative endometrial biopsy, 58 (16.2%) were upgraded based on a final pathology report from hysterectomy specimens. For grade 1, the preoperative endometrial biopsy showed a sensitivity of 80.4%, a specificity of 78.6%, a positive predictive value (PPV) of 83.8% and a negative predictive value (NPV) of 74.5%. Of the 672 patients with the endometrioid cell type on preoperative biopsy, 46 (5.6%) showed a different cell type on final pathology. For the endometrioid cell type, preoperative endometrial biopsy had a sensitivity of 91.3%, a specificity of 64.9%, a PPV of 93.2% and an NPV of 58.6%. This weak predictive value should be considered when selecting patients for conservative hormone treatment.
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Carcinoma Endometrioide/patología , Neoplasias Endometriales/patología , Endometrio/patología , Biopsia/estadística & datos numéricos , Femenino , Humanos , Cuidados Preoperatorios , Estudios RetrospectivosRESUMEN
BACKGROUND: The objective of this study is to construct a preoperative nomogram predicting lymph node metastasis (LNM) in early-cervical cancer patients. METHODS: Between 2009 and 2012, 493 early-cervical cancer patients received hysterectomy and pelvic/para-aortic lymphadenectomy. Patients who were diagnosed during 2009-2010 were assigned to a model-development cohort (n=304) and the others were assigned to a validation cohort (n=189). A multivariate logistic model was created from preoperative clinicopathologic data, from which a nomogram was developed and validated. A predicted probability of LNM<5% was defined as low risk. RESULTS: Age, tumour size assessed by magnetic resonance imaging, and LNM assessed by positron emission tomography/computed tomography were independent predictors of nodal metastasis. The nomogram incorporating these three predictors demonstrated good discrimination and calibration (concordance index=0.878; 95% confidence interval (CI), 0.833-0.917). In the validation cohort, the discrimination accuracy was 0.825 (95% CI, 0.736-0.895). In the model-development cohort, 34% of them were classified as low risk and negative predictive value (NPV) was 99.0%. In the validation cohort, 38% were identified as low risk and NPV was 95.8%. Integrating the model-development and validation cohorts, negative likelihood ratio was 0.094 (95% CI, 0.036-0.248). CONCLUSION: A robust nomogram predicting LNM in early cervical cancer was developed. This model may improve clinical trial design and help physicians to decide whether lymphadenectomy should be performed.
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Ganglios Linfáticos/patología , Nomogramas , Neoplasias del Cuello Uterino/patología , Adulto , Anciano , Estudios de Cohortes , Femenino , Humanos , Modelos Logísticos , Ganglios Linfáticos/cirugía , Metástasis Linfática , Imagen por Resonancia Magnética , Persona de Mediana Edad , Imagen Multimodal , Análisis Multivariante , Estadificación de Neoplasias , Tomografía de Emisión de Positrones , Valor Predictivo de las Pruebas , Neoplasias del Cuello Uterino/cirugía , Adulto JovenRESUMEN
E2F-1 is a downstream regulator of the Rb pathway and is a transcription factor that plays a key role in the control of cell cycle progression. Deregulation of E2F-1 expression and Rb pathway is involved in carcinogenesis. The aim of this study was to evaluate E2F-1 expression and Rb pathway alteration and to elucidate their correlation with clinical and pathologic parameters in epithelial ovarian cancer (EOC). We investigated overexpression of E2F-1 and alterations of p16(INK4a), cyclin D1, CDK4, and pRb using immunohistochemistry and tissue microarray methods in 72 EOC patients. Overexpression of E2F-1 was detected in 45.8% of samples. The overall abnormal expression frequencies of p16(INK4a), cyclin D1, CDK4, and pRb were 33.3%, 11.1%, 12.5%, and 38.9%, respectively. E2F-1 overexpression was not associated with alteration of the Rb pathway. E2F-1 overexpression was correlated with FIGO stage, histologic grade, and mitotic index; it was a valuable prognostic variable along with FIGO stage in the multivariated analysis. The results suggest that E2F-1 has a growth-promoting effect in EOC and that E2F-1 overexpression may provide a useful prognostic indicator for EOC.
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Factor de Transcripción E2F1/genética , Regulación Neoplásica de la Expresión Génica , Predisposición Genética a la Enfermedad , Neoplasias Glandulares y Epiteliales/genética , Neoplasias Ováricas/genética , Distribución de Chi-Cuadrado , Intervalos de Confianza , Ciclina D1/genética , Ciclina D1/metabolismo , Quinasa 4 Dependiente de la Ciclina/genética , Quinasa 4 Dependiente de la Ciclina/metabolismo , Factor de Transcripción E2F1/metabolismo , Femenino , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Análisis Multivariante , Neoplasias Glandulares y Epiteliales/mortalidad , Neoplasias Glandulares y Epiteliales/patología , Neoplasias Ováricas/mortalidad , Neoplasias Ováricas/patología , Probabilidad , Pronóstico , Estudios Retrospectivos , Medición de Riesgo , Sensibilidad y Especificidad , Análisis de Supervivencia , Técnicas de Cultivo de TejidosRESUMEN
OBJECTIVES: Patient-specific (PS) implantation surgical technology has been introduced in recent years and a gradual increase in the associated number of surgical cases has been observed. PS technology uses a patient's own geometry in designing a medical device to provide minimal bone resection with improvement in the prosthetic bone coverage. However, whether PS unicompartmental knee arthroplasty (UKA) provides a better biomechanical effect than standard off-the-shelf prostheses for UKA has not yet been determined, and still remains controversial in both biomechanical and clinical fields. Therefore, the aim of this study was to compare the biomechanical effect between PS and standard off-the-shelf prostheses for UKA. METHODS: The contact stresses on the polyethylene (PE) insert, articular cartilage and lateral meniscus were evaluated in PS and standard off-the-shelf prostheses for UKA using a validated finite element model. Gait cycle loading was applied to evaluate the biomechanical effect in the PS and standard UKAs. RESULTS: The contact stresses on the PE insert were similar for both the PS and standard UKAs. Compared with the standard UKA, the PS UKA did not show any biomechanical effect on the medial PE insert. However, the contact stresses on the articular cartilage and the meniscus in the lateral compartment following the PS UKA exhibited closer values to the healthy knee joint compared with the standard UKA. CONCLUSION: The PS UKA provided mechanics closer to those of the normal knee joint. The decreased contact stress on the opposite compartment may reduce the overall risk of progressive osteoarthritis.Cite this article: K-T. Kang, J. Son, D-S. Suh, S. K. Kwon, O-R. Kwon, Y-G. Koh. Patient-specific medial unicompartmental knee arthroplasty has a greater protective effect on articular cartilage in the lateral compartment: A Finite Element Analysis. Bone Joint Res 2018;7:20-27. DOI: 10.1302/2046-3758.71.BJR-2017-0115.R2.
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OBJECTIVES: Malalignment of the tibial component could influence the long-term survival of a total knee arthroplasty (TKA). The object of this study was to investigate the biomechanical effect of varus and valgus malalignment on the tibial component under stance-phase gait cycle loading conditions. METHODS: Validated finite element models for varus and valgus malalignment by 3° and 5° were developed to evaluate the effect of malalignment on the tibial component in TKA. Maximum contact stress and contact area on a polyethylene insert, maximum contact stress on patellar button and the collateral ligament force were investigated. RESULTS: There was greater total contact stress in the varus alignment than in the valgus, with more marked difference on the medial side. An increase in ligament force was clearly demonstrated, especially in the valgus alignment and force exerted on the medial collateral ligament also increased. CONCLUSION: These results highlight the importance of accurate surgical reconstruction of the coronal tibial alignment of the knee joint. Varus and valgus alignments will influence wear and ligament stability, respectively in TKA.Cite this article: D-S. Suh, K-T. Kang, J. Son, O-R. Kwon, C. Baek, Y-G. Koh. Computational study on the effect of malalignment of the tibial component on the biomechanics of total knee arthroplasty: A Finite Element Analysis. Bone Joint Res 2017;6:623-630. DOI: 10.1302/2046-3758.611.BJR-2016-0088.R2.
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The expression of immediate early response 3 (IER3), a protein with a short half-life, is rapidly induced by various cellular stimuli. We recently reported that IER3 induces the apoptosis of cervical cancer cells and that its expression is downregulated in patients with cervical cancer. However, the molecular mechanism involved in the rapid degradation of IER3 remains unknown. Here, we demonstrate that MDM2 is an E3 ligase that interacts with IER3 and promotes its ubiquitination, followed by proteasomal degradation. Polyubiquitination of the conserved lysine 60 of IER3 is essential for its degradation. In addition, four and a half LIM domains protein 2 (FHL2) binds to both IER3 and MDM2, allowing for efficient MDM2-mediated IER3 degradation by facilitating an association between MDM2 and IER3. Moreover, IER3 induces cell cycle arrest in cervical cancer cells and its activity is further enhanced in cells in which FHL2 or MDM2 was silenced, thereby preventing IER3 degradation. The E6 and E7 oncoproteins of human papilloma virus 18 regulated IER3 expression. FHL2 expression was significantly higher in the squamous epithelium of cervical carcinoma tissues than in non-cancerous cervical tissues, whereas cervical carcinoma expression of IER3 was downregulated in this region. Thus, we determined the molecular mechanism responsible for IER3 degradation, involving a ternary complex of IER3, MDM2 and FHL2, which may contribute to cervical tumor growth. Furthermore, we demonstrated that FHL2 serves as a scaffold for E3 ligase and its substrate during the ubiquitination reaction, a function that has not been previously reported for this protein.
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Proteínas Reguladoras de la Apoptosis/genética , Proteínas con Homeodominio LIM/genética , Proteínas de la Membrana/genética , Proteínas Musculares/genética , Proteínas Proto-Oncogénicas c-mdm2/genética , Factores de Transcripción/genética , Neoplasias del Cuello Uterino/genética , Proliferación Celular/genética , Proteínas de Unión al ADN/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Células HeLa , Papillomavirus Humano 18/genética , Papillomavirus Humano 18/patogenicidad , Humanos , Proteínas Oncogénicas Virales/genética , Complejo de la Endopetidasa Proteasomal/genética , Proteolisis , Ubiquitinación , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/virologíaRESUMEN
Eight hundred second chromosomes were extracted from the Ishigakijima population, one of the southernmost populations of Drosophila melanogaster in Japan. Half of them were extracted in Native cytoplasm (P-type), and half in Foreign cytoplasm (M-type). Various population-genetic parameters, including the frequency of lethal-carrying second chromosomes (Q = 0.235 for the Native; 0.218 for the Foreign), the allelism rate of lethal second chromosome (Ic = 0.0217 for the Native; 0.0134 for the Foreign), the homozygous detrimental and lethal loads (D = 0.179 for the Native; 0.270 for the Foreign; L = 0.262 for the Native; 0.240 for the Foreign), the average degree of dominance of mildly deleterious mutations (hE = 0.244 for the Native; 0.208 for the Foreign), and the components of genetic variance for viability [additive (sigma A2) and dominance (sigma D2)](sigma A2 = 0.0187 for the Native; 0.0172 for the Foreign; sigma D2 = 0.0005 for the Native; 0.0009 for the Foreign) were estimated. The data indicate that D was significantly larger and hE was significantly smaller in the Foreign cytoplasm. However, the estimates of additive and dominance variances were not significantly different between the two cytoplasms. The additive genetic variance for viability in the Ishigakijima population was greater than expected on the basis of mutation-selection balance confirming previous studies on papers of D. melanogaster in warm climates.
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Elementos Transponibles de ADN , Drosophila melanogaster/genética , Variación Genética/genética , Hibridación Genética , Animales , Cruzamientos Genéticos , Citoplasma , Drosophila melanogaster/citología , Femenino , Genes Dominantes , Genes Letales , Heterocigoto , Homocigoto , Japón , Masculino , MutagénesisRESUMEN
The insulin-like growth factor (IGF) binding proteins (IGFBPs) are a family of proteins that bind IGF-I and IGF-II and modulate their biological activities. IGFBP-1 is distinctive among the IGFBPs in its rapid regulation in response to metabolic and hormonal changes. The synthetic glucocorticoid, dexamethasone, increases IGFBP-1 mRNA abundance and gene transcription in rat liver and in H4-II-E rat hepatoma cells. A glucocorticoid response element (GRE) located at nucleotide (nt) -91/-77 is required for dexamethasone to stimulate rat IGFBP-1 promoter activity in transient transfection assays in H4-II-E cells. In addition to the GRE, three accessory regulatory sites [a putative hepatocyte nuclear factor-1 (HNF-1) site (nt -62/-50), an insulin-response element (nt -108/-99), and an upstream site (nt -252/-236)] are involved in dexamethasone stimulation under some, but not all, circumstances. The present study begins to address the mechanism by which transcription factors bound to the putative HNF-1 site act synergistically with the glucocorticoid receptor (GR) bound to the GRE. In gel shift assays, HNF-1alpha and HNF-1beta in H4-II-E extracts bind to the palindromic HNF-1 site. Both half-sites are required. Overexpression of HNF-1beta enhances dexamethasone-stimulated promoter activity. Both the HNF-1 site and the GRE must be intact for stimulation to occur. By contrast, overexpression of HNF-1alpha does not enhance dexamethasone-stimulated promoter activity, although, as also observed with overexpression of HNF-1beta, it inhibits basal promoter activity. Thus, the synergistic effects of HNF-1beta and the GR on dexamethasone-stimulated promoter activity require that they are bound to the HNF-1 site and the GRE, respectively, and may involve protein-protein interactions between the transcription factors, or between them and the basal transcription machinery or a steroid receptor coactivator. Synergy between the ubiquitously expressed GR and HNF-1, which is developmentally regulated and expressed in a limited number of tissues, provides a possible mechanism for tissue- and development-specific regulation of glucocorticoid action.
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Proteínas Portadoras/genética , Proteínas de Unión al ADN , Proteínas Nucleares , Proteínas Gestacionales/genética , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/fisiología , Factores de Transcripción/genética , Factores de Transcripción/fisiología , Activación Transcripcional/efectos de los fármacos , Animales , Proteínas Portadoras/efectos de los fármacos , Núcleo Celular/genética , Núcleo Celular/metabolismo , Dexametasona/farmacología , Sinergismo Farmacológico , Electroforesis en Gel de Poliacrilamida , Factor Nuclear 1 del Hepatocito , Factor Nuclear 1-alfa del Hepatocito , Factor Nuclear 1-beta del Hepatocito , Proteínas Gestacionales/efectos de los fármacos , Regiones Promotoras Genéticas/efectos de los fármacos , Unión Proteica/genética , Ratas , Factores de Transcripción/biosíntesis , Células Tumorales CultivadasRESUMEN
Insulin-like growth factor-binding protein-1 (IGFBP-1) modulates the action of IGFs on target cells. IGFBP-1 transcription is highly regulated by hormonal and metabolic factors. In rat H4-II-E hepatoma cells, IGFBP-1 messenger RNA is stimulated by dexamethasone, cAMP, and phorbol esters, and dominantly inhibited by insulin. To identify the cis-elements that determine transcriptional regulation by these agents, we have coupled rat IGFBP-1 promoter fragments to a luciferase reporter gene and transfected H4-II-E cells using the cationic liposome procedure. Promoter fragments whose 5'-end was at nucleotide (nt) -925 or -327 (with respect to the transcription initiation site, 1) conferred positive regulation of promoter activity by dexamethasone, cAMP, and phorbol esters. Insulin inhibited promoter activity in the presence of any of the three stimulatory agents. Stimulation by cAMP or phorbol esters was abolished when the region between nt -327 and -235 was deleted. Although this region contains potential activating protein-2 and activating protein-1 sites, the sites responsible for this regulation have not yet been identified. By contrast, stimulation by dexamethasone was retained in deletion constructs whose 5'-end was at nt -92, but was abolished by site mutagenesis of either the left or right half-sites of a potential glucocorticoid response element (GRE) located between nt -91 and -77. Surprisingly, substitution mutations in an up-stream region, -108 to -99 (M4), also decreased dexamethasone-stimulated promoter activity despite the presence of an intact GRE. We postulate that a positive factor that binds to the wild-type M4 region neutralizes factors that inhibit interaction of the glucocorticoid receptor with the GRE. The M4 region also is involved in inhibition by insulin. Insulin inhibition of dexamethasone-stimulated promoter activity was lost after deletion of nt -135 to -92 or mutation of the region between nt -108 and -99. This insulin response element is conserved in the human IGFBP-1 promoter and is homologous to the insulin response element of the phosphoenolpyruvate carboxykinase gene, which also is rapidly inhibited by insulin in H4-II-E cells. The rat IGFBP-1 promoter provides a valuable model system for studying the multihormonal regulation of transcription.
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Proteínas Portadoras/antagonistas & inhibidores , Proteínas Portadoras/fisiología , AMP Cíclico/farmacología , Dexametasona/farmacología , Elementos de Facilitación Genéticos/fisiología , Insulina/farmacología , Ésteres del Forbol/farmacología , Animales , Secuencia de Bases , Proteínas Portadoras/genética , ADN/análisis , ADN/genética , Elementos de Facilitación Genéticos/genética , Eliminación de Gen , Regulación de la Expresión Génica/efectos de los fármacos , Genes Reporteros , Glucocorticoides/farmacología , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina , Neoplasias Hepáticas Experimentales/metabolismo , Neoplasias Hepáticas Experimentales/patología , Neoplasias Hepáticas Experimentales/fisiopatología , Datos de Secuencia Molecular , Mutación , Nucleótidos/análisis , Nucleótidos/fisiología , Regiones Promotoras Genéticas/genética , Regiones Promotoras Genéticas/fisiología , ARN Mensajero/análisis , ARN Mensajero/genética , Ratas , Transcripción Genética , Células Tumorales CultivadasRESUMEN
Insulin-like growth factor binding protein-1 (IGFBP-1) modulates the mitogenic actions of IGF-I and IGF-II. Dexamethasone increases IGFBP-1 mRNA abundance and gene transcription in rat liver and in H4-II-E rat hepatoma cells. A glucocorticoid response element (GRE) located at nucleotide (nt) -91/-77 is required for dexamethasone to stimulate rat IGFBP-1 promoter activity in transient transfection assays in H4-II-E cells. Mutagenesis of nt -108/-99 (the M4 region of the insulin response element), however, decreased dexamethasone-stimulated promoter activity despite the presence of an intact GRE, suggesting that regulatory sites in addition to the GRE were required for optimal dexamethasone stimulation. To identify these sites, we introduced 5'-deletion and substitution mutations into rat IGFBP-1 promoter fragments coupled to a luciferase reporter gene and transfected these constructs into H4-II-E cells. Three sites are required for optimal basal promoter activity: a site (nt -62/-50) that binds the liver-enriched transcription factor, hepatocyte nuclear factor-1 (HNF-1), the M4 site, and a putative binding site for transcription factor AP-2 (nt -293/-286). The HNF-1 and M4 sites and an upstream site (nt -252/-236) are also involved in dexamethasone stimulation under some, but not all, circumstances. Mutation of either the HNF-1 site or the M4 site decreased dexamethasone stimulation by more than 80% in constructs whose 5'-end was at nt -92, -135, or -235 but not if the 5' -end was at nt -278 or -327. These results suggest that the nt -278/-236 region can compensate for the loss of the HNF-1 site or the M4 site but that the HNF-1 and M4 sites do not compensate for each other in constructs whose 5'-end was at nt -135 or -235, which lack the nt -278/-236 region. The site within the nt -278/-236 region was localized to nt -252/-236 by deoxyribonuclease I protection and transfection assays. Thus, several cis-elements in the rat IGFBP-1 promoter cooperate, in varying combinations, with the low-affinity GRE to allow optimal dexamethasone-stimulated promoter activity.
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Dexametasona/farmacología , Elementos de Facilitación Genéticos , Glucocorticoides/farmacología , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Regiones Promotoras Genéticas/genética , Transducción de Señal/efectos de los fármacos , Animales , Línea Celular , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/genética , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Ratas , Transcripción GenéticaRESUMEN
After i.v. injection into adult rats, human insulin-like growth factor-binding protein-3 (hIGFBP-3) forms 150-kDa complexes with excess endogenous rat acid-labile subunit (ALS) within 2 min (Lewitt et al., 1993, Endocrinology 133:1797). Because their previous in vitro studies indicated that hIGFBP-3 only bound to ALS in the presence of IGF-I, and because little free IGF-I is present in plasma, the authors postulated that IGF-I had been mobilized to the circulation to saturate the 150-kDa hIGFBP-3 complexes. We examined this hypothesis by determining whether the hIGFBP-3 that appears in 150-kDa complexes 2 min after i.v. injection is accompanied by an increase in IGF-I. Within 2 min, some of the injected hIGFBP-3 (approximately 30% as much as endogenous intact rat IGFBP-3) is present in complexes that are cleared slowly from the circulation and presumed to be 150-kDa complexes. Gel filtration and immunoprecipitation studies performed on blood collected 2 min after injection confirmed that the injected hIGFBP-3 (46-82% as much as rat IGFBP-3) was associated with ALS in 150-kDa complexes. The formation of 150-kDa complexes containing hIGFBP-3 was not accompanied by a corresponding change in the IGF-I content (determined by RIA) of whole serum or 150-kDa serum fractions, suggesting that the hIGFBP-3 had rapidly associated with rALS in vivo without mobilizing IGF-I. Surprisingly, however, hIGFBP-3 was cleared much more rapidly from 150-kDa complexes formed after injection of hIGFBP-3 than after injection of hIGFBP-3:IGF-I complexes, suggesting that the complexes observed after hIGFBP-3 injection might not have formed in vivo. In fact, 150-kDa complexes formed to a similar extent when hIGFBP-3 was added ex vivo to blood collected from rats that had not received hIGFBP-3. We conclude that hIGFBP-3 can rapidly associate with rALS to form 150-kDa complexes in vivo without the mobilization of IGF-I. Because 150-kDa complexes also are formed ex vivo, however, we are unable to resolve whether the complexes that formed in vivo formed as binary or ternary complexes.
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Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/farmacología , Factor I del Crecimiento Similar a la Insulina/química , Animales , Proteínas Portadoras/metabolismo , Cromatografía en Gel , Glicoproteínas/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/química , Cinética , Peso Molecular , Conformación Proteica , Conejos , Radioinmunoensayo , Ratas , Somatomedinas/metabolismoRESUMEN
Nitric oxide (NO) is believed to play an important, but as yet undefined, role in regulating hypoxia inducible gene expression. Recently, we have reported evidence suggesting that the human insulin-like growth factor-binding protein-1 (IGFBP-1) gene is directly regulated by hypoxia through the hypoxia-inducible factor-1 pathway. The goal of the current study was to investigate NO regulation of hypoxic induction of IGFBP-1 gene expression using HepG2 cells, a model system of hepatic gene expression. We report that a NO generator, sodium nitroprusside, significantly diminishes hypoxic activation of IGFBP-1 protein and messenger ribonucleic acid expression. Furthermore, these effects are independent of guanylate cyclase/ cGMP signaling, as two different inhibitors, LY 83583, a specific inhibitor of guanylate cyclase, and KT 5823, a protein kinase G inhibitor, had no effect on IGFBP-1 induction by hypoxia. Hypoxic induction of a reporter gene containing four tandemly ligated hypoxia response elements was completely blocked by sodium nitroprusside, but not by 8-bromo-cGMP, an analog ofcGMP. These results suggest that NO blocks hypoxic induction of IGFBP-1 by a guanylate cyclase/ cGMP-independent pathway, possibly at the level of oxygen sensing. The impaired hypoxia regulation of IGFBP-1 by nitric oxide may play a key role in the hyperinduction of IGFBP-1 observed in pathophysiological conditions such as fetal hypoxia and preeclampsia where dysregulation of NO has been observed.
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Carbazoles , Hipoxia de la Célula , Regulación de la Expresión Génica , Indoles , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Óxido Nítrico/fisiología , Nitroprusiato/farmacología , Alcaloides/farmacología , Aminoquinolinas/farmacología , Secuencia de Bases , Sitios de Unión , Carcinoma Hepatocelular , Secuencia Conservada , GMP Cíclico/análogos & derivados , GMP Cíclico/farmacología , GMP Cíclico/fisiología , Proteínas de Unión al ADN/metabolismo , Inhibidores Enzimáticos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Guanilato Ciclasa/metabolismo , Humanos , Factor 1 Inducible por Hipoxia , Subunidad alfa del Factor 1 Inducible por Hipoxia , Cinética , Hígado/metabolismo , Neoplasias Hepáticas , Luciferasas/genética , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Biosíntesis de Proteínas , Proteínas Recombinantes de Fusión/biosíntesis , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/metabolismo , Transcripción Genética , Transfección , Células Tumorales CultivadasRESUMEN
Three different strains of Aureobasidium pullulans were grown in batch cultures to compare their abilities for the production of fructo-oligosaccharides. Specific intracellular enzyme activity was the highest with strain KCCM 12017 and enzyme production was closely coupled to growth. Using A. pullulans cells, 166 g/l fructo-oligosaccharides was produced from 360 g/l molasses sugar as sucrose equivalent at 55 degrees C and pH 5.5 after 24 h incubation.
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Agricultura/métodos , Ascomicetos/metabolismo , Fructanos/biosíntesis , Melaza/microbiología , Oligosacáridos/biosíntesis , Agricultura/economía , Alimentación Animal , Cromatografía Líquida de Alta Presión , Hexosiltransferasas/metabolismo , TemperaturaRESUMEN
This study was conducted to determine the optimal temperature and time for the regiospecific oxidation of primary alcohol groups in corn starch with 2,2,6,6-tetramethyl-1-piperidinyl oxoammonium ion (TEMPO) and sodium hyphochlorite (NaOCl). The study also elucidated the molecular structure of fully oxidized corn starch (FOCS) prepared at optimum temperature and physicochemical properties of the partially (10%, 20%, and 30%) oxidized corn starches (POCS). The reaction time rapidly decreased up to 30 degrees C, and then gradually decreased. Selectivity, yield, and viscosity were drastically reduced at temperatures higher than 35 to 40 degrees C. Optimal oxidation temperature for the production of FOCS was determined as 35 degrees C. Regiospecific oxidation of the primary alcohol group without oxidation of the secondary alcohol group was confirmed in (13)C-NMR and IR spectra. Water-binding capacity, swelling power, solubility power, and transmittance of POCS increased as the degree of oxidation increased. Hardness, gumminess, and chewiness of corn starch gel containing POCS were not significantly different from those of native corn starch gel at 1-d storage, but the values of the starch gel containing POCS were smaller than those of the native starch gel after 1-d storage. However, springiness and cohesiveness did not differ significantly among the samples regardless of storage time.
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Antioxidantes/química , Óxidos N-Cíclicos/química , Conservación de Alimentos/métodos , Hipoclorito de Sodio/química , Almidón/química , Isótopos de Carbono , Tecnología de Alimentos/métodos , Geles , Espectroscopía de Resonancia Magnética , Estructura Molecular , Oxidación-Reducción , Solubilidad , Relación Estructura-Actividad , Temperatura , Factores de Tiempo , Viscosidad , Zea mays/químicaRESUMEN
This study examined the characteristics of the oxidation reaction on the primary alcohol groups in cellulose involving the 2,2,6,6-tetramethyl-1-piperidinyl oxoammonium ion (TEMPO) and determined the optimum conditions for the preparation of oxidized cellulose (OC). The applicability of OC in polysaccharide systems was also investigated. The effects of TEMPO, sodium bromide (NaBr), and temperature on the oxidation reaction time, yield, and selectivity for primary alcohol groups were examined using response surface methodology (RSM). The reaction time decreased with increases in the temperature and the levels of TEMPO and NaBr. The yield increased with the level of NaBr and decreased as the temperature increased. Selectivity increased with the temperature and decreased as the levels of TEMPO and NaBr increased. The optimum levels of TEMPO and NaBr and the optimum temperature for the production of OC were determined as 0.3 mM/100 mM anhydroglucose unit (AGU), 50 mM/100 mM AGU, and 25 degrees C, respectively. The water and oil binding capacity and viscosity of cellulose increased with oxidation. Wheat starch containing OC exhibited a decreased initial pasting temperature and setback, but increased peak viscosity, gelatinization, and retrogradation enthalpy (DeltaH). The hardness of the wheat starch gel decreased significantly upon the addition of OC.
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Antioxidantes/química , Celulosa/química , Óxidos N-Cíclicos/química , Tecnología de Alimentos/métodos , Bromuros/química , Relación Dosis-Respuesta a Droga , Estructura Molecular , Oxidación-Reducción , Compuestos de Sodio/química , Almidón/química , Relación Estructura-Actividad , Temperatura , Factores de Tiempo , ViscosidadRESUMEN
The purpose of this study was to evaluate and compare the outcomes of laparoscopic surgery with those of conventional abdominal surgery in patients with early endometrial cancer. From 1997 to 2003, 79 patients underwent laparoscopic-assisted vaginal hysterectomy with or without lymphadenectomy. Laparoscopy was performed on patients deemed clinical stage I in preoperative studies. Of the 79 patients, 74 found to be surgical stage I or II were enrolled in the comparative study. As a control group, we selected 168 laparotomy cases at the same disease stage as the laparoscopy group. Operation time, amount of blood transfusion, and hemoglobin changes were similar for both groups. In the laparoscopy group, the number of lymph nodes obtained was significantly higher, and the number of postoperative complications was lower compared to the laparotomy group. The hospital stay was significantly shorter for laparoscopy group. Three-year recurrence-free survival rates were similar, being 97.5% for the laparoscopy group and 98.6% for the laparotomy group. We conclude that laparoscopic surgery for treatment of early endometrial cancer is a safe and effective alternative to laparotomy in terms of perioperative complications. Three-year recurrence-free survival did not differ significantly between the groups. However, long-term survival and risk of recurrence have yet to be determined.
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Neoplasias Endometriales/patología , Neoplasias Endometriales/cirugía , Histerectomía , Laparoscopía , Adulto , Anciano , Anciano de 80 o más Años , Progresión de la Enfermedad , Femenino , Humanos , Persona de Mediana Edad , Estadificación de Neoplasias , Tasa de Supervivencia , Resultado del TratamientoRESUMEN
Segregation of sperm abnormality level and H-2 haplotypes was investigated in F2 hybrid males obtained from reciprocal crosses involving two B10.congenic strains carrying H-2 and the Y chromosome of Japanese wild mice: B10.MOL-OHM (H-2wm4, 23.1% of sperm abnormalities) and B10.MOL-OKB (H-2wm8, 11.1% of sperm abnormalities). In both types of crosses mean levels of abnormal spermatozoa were significantly higher for males typed as H-2wm4/H-2wm4 than for heterozygous H-2wm4/H-2wm8 or homozygous H-2wm8/H-2wm8. These results suggest that the gene for high sperm abnormality is linked to H-2 complex of the B10.MOL-OHM strain.
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Antígenos H-2/genética , Espermatozoides/anomalías , Cromosoma Y , Animales , Masculino , MuridaeRESUMEN
In an isolated population of Drosophila melanogaster on Ishigaki Island the chromosomal distribution of several retrotransposons, including copia, 412, 297, 17.6, I, and jockey elements, was examined by in situ hybridization. In this population the cosmopolitan inversion, In(2L)t, is known to exist in high frequency. One major haplotype concerning the occupied sites of the transposable elements was identified in the In(2L)t-carrying chromosomes. This haplotype is suggested to be the ancestral one. The age of the inversion in this local population was estimated to be 1,400 generations. The transposition rates of these elements were estimated based on the age of the inversion and the number of the elements lost and gained. The excision rates were in the range from 9.13 x 10(-5) to 2.25 x 10(-4) per site per generation. They were similar each other in the copia-like elements as well as in the LINE-like elements. The rate was higher in the copia-like elements than in the LINE-like elements. Insertions occurred in the range from 6.79 x 10(-4) to 9.05 x 10(-4) per element per generation. It is herein shown that both insertions and excisions occurred at a significantly higher rate in this population than in the laboratory.