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Laser plasma-based accelerators provide an excellent source of collimated, bright, and adequately coherent betatron-type x-ray pulses with potential applications in science and industry. So far the laser plasma-based betatron radiation has been described within the concept of classical Liénard-Wiechert potentials incorporated in particle-in-cell simulations, a computing power-demanding approach, especially for the case of multi-petawatt lasers. In this work, we describe the laser plasma-based generation of betatron radiation at the most fundamental level of quantum mechanics. In our approach, photon emission from the relativistic electrons in the plasma bubble is described within a nonlinear quantum electrodynamics (QED) framework. The reported QED-based betatron radiation results are in excellent agreement with similar results using Liénard-Wiechert potentials, as well as in very good agreement with betatron radiation measurements, obtained with multi-10-TW lasers interacting with He and multielectron N[Formula: see text] gas targets. Furthermore, our QED approach results in a dramatic reduction of the computational runtime demands, making it a favorable tool for designing betatron radiation experiments, especially in multi-petawatt laser facilities.
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Laser WakeField Acceleration (LWFA) is extensively used as a high-energy electron source, with electrons achieving energies up to the GeV level. The produced electron beam characteristics depend strongly on the gas density profile. When the gaseous target is a gas jet, the gas density profile is affected by parameters, such as the nozzle geometry, the gas used, and the backing pressure applied to the gas valve. An electron source based on the LWFA mechanism has recently been developed at the Institute of Plasma Physics and Lasers. To improve controllability over the electron source, we developed a set of 3D-printed nozzles suitable for creating different gas density profiles according to the experimental necessities. Here, we present a study of the design, manufacturing, evaluation, and performance of a 3D-printed nozzle intended for LWFA experiments.
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The temporal rearrangement of the spectral components of an ultrafast and intense laser pulse, i.e., the chirp of the pulse, offers significant possibilities for controlling its interaction with matter and plasma. In the propagation of ultra-strong laser pulses within the self-induced plasma, laser pulse chirp can play a major role in the dynamics of wakefield and plasma bubble formation, as well as in the electron injection and related electron acceleration. Here, we experimentally demonstrate the control of the generation efficiency of a relativistic electron beam, with respect to maximum electron energy and current, by accurately varying the chirp value of a multi-10-TW laser pulse. We explicitly show that positively chirped laser pulses, i.e., pulses with instantaneous frequency increasing with time, accelerate electrons in the order of 100 MeV much more efficiently in comparison to unchirped or negatively chirped pulses. Corresponding Particle-In-Cell simulations strongly support the experimental results, depicting a smoother plasma bubble density distribution and electron injection conditions that favor the maximum acceleration of the electron beam, when positively chirped laser pulses are used. Our results, aside from extending the validity of similar studies reported for PW laser pulses, provide the ground for understanding the subtle dynamics of an efficient plasma electron accelerator driven by chirped laser pulses.
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BACKGROUND: In the TNT trial of triple negative breast cancer (NCT00532727), germline BRCA1/2 mutations were present in 28% of carboplatin responders. We assessed quantitative measures of structural chromosomal instability (CIN) to identify a wider patient subgroup within TNT with preferential benefit from carboplatin over docetaxel. PATIENTS AND METHODS: Copy number aberrations (CNAs) were established from 135 formalin-fixed paraffin-embedded primary carcinomas using Illumina OmniExpress SNP-arrays. Seven published [allelic imbalanced CNA (AiCNA); allelic balanced CNA (AbCNA); copy number neutral loss of heterozygosity (CnLOH); number of telomeric allelic imbalances (NtAI); BRCA1-like status; percentage of genome altered (PGA); homologous recombination deficiency (HRD) scores] and two novel [Shannon diversity index (SI); high-level amplifications (HLAMP)] CIN-measurements were derived. HLAMP was defined based on the presence of at least one of the top 5% amplified cytobands located on 1q, 8q and 10p. Continuous CIN-measurements were divided into tertiles. All nine CIN-measurements were used to analyse objective response rate (ORR) and progression-free survival (PFS). RESULTS: Patients with tumours without HLAMP had a numerically higher ORR and significantly longer PFS in the carboplatin (C) than in the docetaxel (D) arm [56% (C) versus 29% (D), PHLAMP,quiet = 0.085; PFS 6.1 months (C) versus 4.1 months (D), Pinteraction/HLAMP = 0.047]. In the carboplatin arm, patients with tumours showing intermediate telomeric NtAI and AiCNA had higher ORR [54% (C) versus 20% (D), PNtAI,intermediate = 0.03; 62% (C) versus 33% (D), PAiCNA,intermediate = 0.076]. Patients with high AiCNA and PGA had shorter PFS in the carboplatin arm [3.4 months (high) versus 5.7 months (low/intermediate); and 3.8 months (high) versus 5.6 months (low/intermediate), respectively; Pinteraction/AiCNA = 0.027, Padj.interaction/AiCNA = 0.125 and Pinteraction/PGA = 0.053, Padj.interaction/PGA = 0.176], whilst no difference was observed in the docetaxel arm. CONCLUSIONS: Patients with tumours lacking HLAMP and demonstrating intermediate CIN-measurements formed a subgroup benefitting from carboplatin relative to docetaxel treatment within the TNT trial. This suggests a complex and paradoxical relationship between the extent of genomic instability in primary tumours and treatment response in the metastatic setting.
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Protocolos de Quimioterapia Combinada Antineoplásica , Neoplasias de Mama Triplo Negativas , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores , Carboplatina/uso terapêutico , Instabilidade Cromossômica/genética , Humanos , Fenótipo , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/genéticaRESUMO
Mastication is a complex sensorimotor interaction between the central nervous system and the peripheral masticatory apparatus. To understand the effect of oro-facial abnormalities on mastication, it is important to first understand the normal development of jaw sensorimotor control and chewing in healthy children. Original studies which investigated four main objective parameters of chewing, i.e. maximum occlusal bite force, electromyography (EMG), jaw kinematics and chewing efficiency in children were systematically searched using three established databases. The targeted sample was healthy children below the age of 18-years. All studies that subjectively assessed mastication, studies of children with abnormalities, or non-English studies were excluded. A total of 6193 papers were identified, 53 met the final inclusion criteria. Results are presented according to the dentition stage. Children below 6-years (primary dentition) had lower biting forces and EMG activity, and the frontal jaw movement pattern was more laterally displaced and less stable than children older than 6-years. EMG activities and bite forces increased in children 6- to 10-year-old (early mixed dentition) with a reduction in lateral jaw displacement and an increase in vertical jaw displacement. Twelve-year-old children were able to chew food into smaller particles compared to 6-year-olds. Gender differences were visible in all parameters except EMG activity in late mixed dentition (10- to 12-years). After 12-years, there was a significant increase in bite forces and EMG activities, and the frontal jaw pattern became similar to adults. Studied chewing parameters gradually improve with the development of the oro-facial structures and were mainly influenced by dental eruption. A significant development of chewing parameters occurs after 12â¯years of age. A transition to the adult-type of masticatory behavior occurs between 10- to 14-years of age.
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Envelhecimento/fisiologia , Arcada Osseodentária/fisiologia , Mastigação/fisiologia , Humanos , Desenvolvimento MaxilofacialRESUMO
Tooth loss, decreased mass and strength of the masticatory muscles leading to difficulty in chewing have been suggested as important determinants of eating and nutrition in the elderly. To compensate for the loss of teeth, in particular, a majority of the elderly rely on dental prosthesis for chewing. Chewing function is indeed an important aspect of oral health, and therefore, oral rehabilitation procedures should aim to restore or maintain adequate function. However, even if the possibilities to anatomically restore lost teeth and occlusion have never been better; conventional rehabilitation procedures may still fail to optimally restore oral functions. Perhaps this is due to the lack of focus on the importance of the brain in the rehabilitation procedures. Therefore, the aim of this narrative review was to discuss the importance of maintaining or restoring optimum chewing function in the superageing population and to summarise the emerging studies on oral motor task performance and measures of cortical neuroplasticity induced by systematic training paradigms in healthy participants. Further, brain imaging studies in patients undergoing or undergone oral rehabilitation procedures will be discussed. Overall, this information is believed to enhance the understanding and develop better rehabilitative strategies to exploit training-induced cortical neuroplasticity in individuals affected by impaired oral motor coordination and function. Training or relearning of oral motor tasks could be important to optimise masticatory performance in dental prosthesis users and may represent a much-needed paradigm shift in the approach to oral rehabilitation procedures.
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Adaptação Fisiológica/fisiologia , Mastigação/fisiologia , Plasticidade Neuronal/fisiologia , Salivação/fisiologia , Córtex Somatossensorial/fisiopatologia , Perda de Dente/fisiopatologia , Força de Mordida , Prótese Dentária , Humanos , Saúde Bucal , Perda de Dente/psicologiaRESUMO
BACKGROUND: VS38c is a monoclonal antibody that recognises a rough endoplasmic reticulum (rER) intracellular antigen termed cytoskeleton-linking membrane protein 63. rER is typically found in viable tumour cells and is abundant in osteosarcoma cells. The aim of this study was to determine the diagnostic and prognostic utility of VS38c in the histological assessment of osteosarcoma and other bone tumours/tumour-like leisons. METHODS: Immunohistochemical staining with VS38c was carried out on formalin-fixed specimens of osteosarcoma (pre/post-chemotherapy) and a wide range of benign and malignant bone lesions. In addition, VS38c staining of cultures of MG63 and Sa0S2 osteosarcoma cell cultures. (±cisplatin and actinomycin D-treatment) was analysed. RESULTS: VS38c strongly stained tumour cells in all low-grade and high-grade osteosarcomas and in undifferentiated sarcomas and high-grade chondrosarcomas. There was little or no VS38c staining of low-grade chondrosarcomas or chordomas and variable staining of Ewing sarcomas. Osteoblasts in benign bone-forming tumours and mononuclear stromal cells in chondroblastomas, giant cell tumours and non-ossifying fibromas strongly stained for VS38c. VS38c staining was absent in cisplatin and actinomycin D treated Sa0S2 and MG63 cells. In specimens of osteosarcoma post-neoadjuvant therapy, VS38c staining was absent in most morphologically necrotic areas of tumor although some cells with pyknotic nuclei stained for VS38c in these areas. Most tumour cells exhibiting atypical nuclear forms were not stained by VS38c. CONCLUSIONS: Our findings show that VS38c is a sensitive but not specific diagnostic marker of osteosarcoma. Staining with VS38c identifies viable osteosarcoma cells, a feature which may be useful in the assessment of percentage tumour necrosis post-neoadjuvant chemotherapy.
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Aberrant Wnt signaling within breast cancer is associated with poor prognosis, but regulation of this pathway in breast tissue remains poorly understood and the consequences of immediate or long-term dysregulation remain elusive. The exact contribution of the Wnt-regulating proteins adenomatous polyposis coli (APC) and APC2 in the pathogenesis of human breast cancer are ill-defined, but our analysis of publically available array data sets indicates that tumors with concomitant low expression of both proteins occurs more frequently in the 'triple negative' phenotype, which is a subtype of breast cancer with particularly poor prognosis. We have used mouse transgenics to delete Apc and/or Apc2 from mouse mammary epithelium to elucidate the significance of these proteins in mammary homeostasis and delineate their influences on Wnt signaling and tumorigenesis. Loss of either protein alone failed to affect Wnt signaling levels or tissue homeostasis. Strikingly, concomitant loss led to local disruption of ß-catenin status, disruption in epithelial integrity, cohesion and polarity, increased cell division and a distinctive form of ductal hyperplasia with 'squamoid' ghost cell nodules in young animals. Upon aging, the development of Wnt activated mammary carcinomas with squamous differentiation was accompanied by a significantly reduced survival. This novel Wnt-driven mammary tumor model highlights the importance of functional redundancies existing between the Apc proteins both in normal homeostasis and in tumorigenesis.
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Proteína da Polipose Adenomatosa do Colo/genética , Proteína da Polipose Adenomatosa do Colo/metabolismo , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Epitélio/metabolismo , Animais , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/mortalidade , Neoplasias da Mama/patologia , Carcinogênese/genética , Carcinogênese/metabolismo , Variações do Número de Cópias de DNA , Epitélio/patologia , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Homeostase/genética , Humanos , Hiperplasia , Lactação/genética , Neoplasias Mamárias Animais , Camundongos , Camundongos Transgênicos , Prognóstico , beta Catenina/genética , beta Catenina/metabolismoRESUMO
OBJECTIVES: To compare a low-tube-voltage with or without high-iodine-load multidetector CT (MDCT) protocol with a normal-tube-voltage, normal-iodine-load (standard) protocol in patients with pancreatic ductal adenocarcinoma (PDAC) with respect to tumour conspicuity and image quality. METHODS: Thirty consecutive patients (mean age: 66 years, men/women: 14/16) preoperatively underwent triple-phase 64-channel MDCT examinations twice according to: (i) 120-kV standard protocol (PS; 0.75 g iodine (I)/kg body weight, n = 30) and (ii) 80-kV protocol A (PA; 0.75 g I/kg, n = 14) or protocol B (PB; 1 g I/kg, n = 16). Two independent readers evaluated tumour delineation and image quality blindly for all protocols. A third reader estimated the pancreas-to-tumour contrast-to-noise ratio (CNR). Statistical analysis was performed with the Chi-square test. RESULTS: Tumour delineation was significantly better in PB and PA compared with PS (P = 0.02). The evaluation of image quality was similar for the three protocols (all, P > 0.05). The highest CNR was observed with PB and was significantly better compared to PA (P = 0.02) and PS (P = 0.0002). CONCLUSION: In patients with PDAC, a low-tube-voltage, high-iodine-load protocol improves tumour delineation and CNR leading to higher tumour conspicuity compared to standard protocol MDCT. KEY POINTS: ⢠Low-tube-voltage high-iodine-load MDCT improves pancreatic cancer conspicuity compared to a standard protocol. ⢠The pancreas-to-tumour attenuation difference increases significantly by reducing the tube voltage. ⢠The radiation exposure dose decreases by reducing the tube voltage.
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Carcinoma Ductal Pancreático/diagnóstico por imagem , Iopamidol/análogos & derivados , Tomografia Computadorizada Multidetectores/métodos , Neoplasias Pancreáticas/diagnóstico por imagem , Intensificação de Imagem Radiográfica/métodos , Ácidos Tri-Iodobenzoicos/farmacocinética , Idoso , Meios de Contraste/farmacocinética , Feminino , Humanos , Iopamidol/farmacocinética , Masculino , Estudos Prospectivos , Doses de Radiação , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Reprodutibilidade dos TestesRESUMO
Osteosarcoma is the most common primary malignancy of the skeleton and is prevalent in children and adolescents. Survival rates are poor and have remained stagnant owing to chemoresistance and the high propensity to form lung metastases. In this study, we used in vivo transgenic models of c-fos oncogene-induced osteosarcoma and chondrosarcoma in addition to c-Fos-inducible systems in vitro to investigate downstream signalling pathways that regulate osteosarcoma growth and metastasis. Fgfr1 (fibroblast growth factor receptor 1) was identified as a novel c-Fos/activator protein-1(AP-1)-regulated gene. Induction of c-Fos in vitro in osteoblasts and chondroblasts caused an increase in Fgfr1 RNA and FGFR1 protein expression levels that resulted in increased and sustained activation of mitogen-activated protein kinases (MAPKs), morphological transformation and increased anchorage-independent growth in response to FGF2 ligand treatment. High levels of FGFR1 protein and activated pFRS2α signalling were observed in murine and human osteosarcomas. Pharmacological inhibition of FGFR1 signalling blocked MAPK activation and colony growth of osteosarcoma cells in vitro. Orthotopic injection in vivo of FGFR1-silenced osteosarcoma cells caused a marked twofold to fivefold decrease in spontaneous lung metastases. Similarly, inhibition of FGFR signalling in vivo with the small-molecule inhibitor AZD4547 markedly reduced the number and size of metastatic nodules. Thus deregulated FGFR signalling has an important role in osteoblast transformation and osteosarcoma formation and regulates the development of lung metastases. Our findings support the development of anti-FGFR inhibitors as potential antimetastatic therapy.
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Neoplasias Pulmonares/secundário , Osteossarcoma/patologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Fator de Transcrição AP-1/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Condrossarcoma/genética , Condrossarcoma/patologia , Colo/efeitos dos fármacos , Colo/patologia , Ativação Enzimática/efeitos dos fármacos , Fatores de Crescimento de Fibroblastos/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Inativação Gênica , Humanos , Masculino , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Oncogenes/genética , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteoblastos/patologia , Osteossarcoma/genética , Proteínas Proto-Oncogênicas c-fos/genética , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/deficiência , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/genética , Transdução de Sinais/efeitos dos fármacos , Ativação TranscricionalRESUMO
c-Fos homozygous mice lack osteoclasts with a failure of the teeth to erupt and with an arrest of root development. Here, we characterize the defects associated with the failure in root development and the loss of the tooth-bone interface, and we investigate the underlying causes. We show that, while homozygous c-Fos mice have no multinucleated osteoclasts, heterozygous mice have a reduction in the number of osteoclasts with a reduction in the tooth-bone interface during development and subtle skeletal defects postnatally. In the homozygous mutants bone is found to penetrate the tooth, particularly at the apical end, physically disrupting the root forming HERS (Hertwig's epithelial root sheath) cells. The cells of the HERS continue to proliferate but cannot extend downward due to the presence of bone, leading to a loss of root formation. Tooth germ culture showed that the developing tooth invaded the static bone in mutant tissue, rather than the bone encroaching on the tooth. Although c-Fos has been shown to be expressed in developing teeth, the defect in maintenance of the tooth-bone interface appears to be driven solely by the lack of osteoclasts, as this defect can be rescued in the presence of donor osteoclasts. The rescue suggests that signals from the tooth recruit osteoclasts to clear the bone from around the tooth, allowing the tooth to grow, form roots, and later erupt.
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Osteoclastos/fisiologia , Proteínas Proto-Oncogênicas c-fos/fisiologia , Erupção Dentária/fisiologia , Raiz Dentária/anormalidades , Animais , Homozigoto , Anormalidades Maxilomandibulares/genética , Anormalidades Maxilomandibulares/fisiopatologia , Desenvolvimento Maxilofacial/genética , Desenvolvimento Maxilofacial/fisiologia , Camundongos , Camundongos Endogâmicos C57BL/genética , Camundongos Mutantes , Proteínas Proto-Oncogênicas c-fos/genética , Erupção Dentária/genética , Raiz Dentária/crescimento & desenvolvimentoRESUMO
Jaw actions adapt to the changing properties of food that occur during a masticatory sequence. In the present study, we investigated how the time-varying activation profile of the masseter muscle changes during natural chewing in humans and how food hardness affects the profile. We recorded surface electromyography (EMG) of the masseter muscle together with the movement of the lower jaw in 14 healthy young adults (mean age 22) when chewing gelatin-based model food of two different hardness. The muscle activity and the jaw kinematics were analysed for different phases of the chewing cycles. The increase in the excitatory drive of the masseter muscle was biphasic during the jaw-closing phase showing early and late components. The transition between these components occurred approximately at the time of tooth-food contact. During the masticatory sequence, when the food was particularised, the size of the early component as well as the peak amplitude of the EMG significantly decreased along with a reduction in the duration of the jaw-closing phase. Except for amplitude scaling, food hardness did not appreciably affect the muscle's activation profile. In conclusion, when chewing food during natural conditions, masseter muscle activation adapted throughout the masticatory sequence, principally during the jaw-closing phase and influenced both early and late muscle activation components. Furthermore, the adaptation of jaw actions to food hardness was affected by amplitude scaling of the magnitude of the muscle activity throughout the masticatory sequence.
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Adaptação Fisiológica/fisiologia , Eletromiografia , Músculo Masseter/fisiologia , Mastigação/fisiologia , Músculo Temporal/fisiologia , Adulto , Fenômenos Biomecânicos , Feminino , Alimentos , Dureza , Humanos , MasculinoRESUMO
OBJECTIVES: To characterize basal differentiation tendencies of a human embryonic stem (hES) cell line, KCL-002. MATERIALS AND METHODS: In vitro specification and differentiation of hES cells were carried out using embryoid body (EB) cultures and tests of pluripotency and in vivo differentiation were performed by teratoma assays in SCID mice. Real-time PCR, immunohistochemistry, flow cytometry and histological analyses were used to identify expression of genes and proteins associated with the ectodermal, endodermal and mesodermal germ layers. RESULTS: Undifferentiated KCL-002 cells expressed characteristic markers of pluripotent stem cells such as Nanog, Sox-2, Oct-4 and TRA 1-60. When differentiated in vitro as EB cultures, expression of pluripotency, endodermal and ectodermal markers decreased rapidly. In contrast, mesodermal and mesenchymal markers such as VEGFR-2, α-actin and vimentin increased during EB differentiation as shown by qPCR, immunostaining and flow cytometric analyses. Teratoma formation in SCID mice demonstrated the potential to form all germ layers in vivo with a greater proportion of the tumours containing mesenchymal derivatives. CONCLUSIONS: The data presented suggest that the KCL-002 hES cell line is pluripotent and harbours a bias in basal differentiation tendencies towards mesodermal and mesenchymal lineage cells. Characterizing innate differentiation propensities of hES cell lines is important for understanding heterogeneity between different cell lines and for further studies aimed at deriving specific lineages from hES cells.
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Células-Tronco Embrionárias/citologia , Mesoderma/citologia , Actinas/genética , Actinas/metabolismo , Animais , Antígenos de Superfície/genética , Antígenos de Superfície/metabolismo , Diferenciação Celular , Linhagem Celular , Ectoderma/metabolismo , Células-Tronco Embrionárias/metabolismo , Endoderma/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Mesoderma/metabolismo , Camundongos , Proteína Homeobox Nanog , Fator 3 de Transcrição de Octâmero/genética , Fator 3 de Transcrição de Octâmero/metabolismo , Proteoglicanas/genética , Proteoglicanas/metabolismo , Fatores de Transcrição SOXB1/genética , Fatores de Transcrição SOXB1/metabolismo , Transplante Heterólogo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
Skeletal growth and homeostasis require the finely orchestrated secretion of mineralized tissue matrices by highly specialized cells, balanced with their degradation by osteoclasts. Time- and site-specific expression of Dlx and Msx homeobox genes in the cells secreting these matrices have been identified as important elements in the regulation of skeletal morphology. Such specific expression patterns have also been reported in osteoclasts for Msx genes. The aim of the present study was to establish the expression patterns of Dlx genes in osteoclasts and identify their function in regulating skeletal morphology. The expression patterns of all Dlx genes were examined during the whole osteoclastogenesis using different in vitro models. The results revealed that Dlx1 and Dlx2 are the only Dlx family members with a possible function in osteoclastogenesis as well as in mature osteoclasts. Dlx5 and Dlx6 were detected in the cultures but appear to be markers of monocytes and their derivatives. In vivo, Dlx2 expression in osteoclasts was examined using a Dlx2/LacZ transgenic mouse. Dlx2 is expressed in a subpopulation of osteoclasts in association with tooth, brain, nerve, and bone marrow volumetric growths. Altogether the present data suggest a role for Dlx2 in regulation of skeletal morphogenesis via functions within osteoclasts.
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Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Família Multigênica/genética , Osteoclastos/metabolismo , Fatores de Transcrição/genética , Fosfatase Ácida/metabolismo , Animais , Diferenciação Celular/genética , Linhagem Celular , Perfilação da Expressão Gênica , Proteínas de Homeodomínio/metabolismo , Isoenzimas/metabolismo , Masculino , Mandíbula/citologia , Mandíbula/enzimologia , Mandíbula/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Osteoclastos/citologia , Osteoclastos/enzimologia , Osteogênese/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fosfatase Ácida Resistente a Tartarato , Fatores de Transcrição/metabolismo , beta-Galactosidase/metabolismoRESUMO
Despite the excellent survival of Wilms tumour patients treated with multimodality therapy, approximately 15% will suffer from tumour relapse, where response rates are markedly reduced. We have carried out microarray-based comparative genomic hybridisation on a series of 76 Wilms tumour samples, enriched for cases which recurred, to identify changes in DNA copy number associated with clinical outcome. Using 1Mb-spaced genome-wide BAC arrays, the most significantly different genomic changes between favourable histology tumours that did (n = 37), and did not (n = 39), subsequently relapse were gains on 1q, and novel deletions at 12q24 and 18q21. Further relapse-associated loci included losses at 1q32.1, 2q36.3-2q37.1, and gain at 13q31. 1q gains correlated strongly with loss of 1p and/or 16q. In 3 of 11 cases with concurrent 1p(-)/1q(+), a breakpoint was identified at 1p13. Multiple low-level sub-megabase gains along the length of 1q were identified using chromosome 1 tiling-path arrays. One such recurrent region at 1q22-q23.1 included candidate genes RAB25, NES, CRABP2, HDGF and NTRK1, which were screened for mRNA expression using quantitative RT-PCR. These data provide a high-resolution catalogue of genomic copy number changes in relapsing favourable histology Wilms tumours.
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Neoplasias Renais/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Tumor de Wilms/genética , Aberrações Cromossômicas , Deleção Cromossômica , Cromossomos Humanos Par 1/genética , Cromossomos Humanos Par 16/genética , Cromossomos Humanos Par 8/genética , DNA de Neoplasias/genética , Genes do Tumor de Wilms/fisiologia , Humanos , Neoplasias Renais/patologia , Recidiva Local de Neoplasia/genética , RNA Mensageiro/análise , RNA Neoplásico/análise , Resultado do Tratamento , Tumor de Wilms/patologiaRESUMO
The RANKL/OPG/RANK pathway is the key mediator of osteoclastogenesis. Mononuclear cells may be implicated in post-menopausal osteoporosis. The effect of estrogen or raloxifene on bone resorption and the expression of RANKL/OPG/RANK in peripheral blood mononuclear cells (PBMCs) was examined. Twenty-nine women with post-menopausal osteoporosis were treated with estrogen (HRT) or raloxifene for 12 months. Bone mineral density (BMD) was measured at baseline and at 12 months at the spine and hip. Serum C-terminal telopeptide (CTX) and OPG were measured at baseline and at 1, 3, 6 and 12 months. PBMCs were isolated from 17 women and changes in RANKL, OPG and RANK mRNA were determined. The effects of estrogen or raloxifene in PBMCs in vitro were also assessed. BMD increased following treatment (lumbar spine % change mean [S.E.M.]: 4.3% [0.9], p<0.001). Serum CTX decreased (6 months: -43.7% [6.0], p<0.0001). Serum OPG declined gradually (12 months: -26.4% [4.4], p<0.001). RANKL, OPG and RANK gene expression decreased (6 months: RANKL 50.0% [24.8] p<0.001, OPG: 21.7% [28] p<0.001, RANK: 76.6% [10.2] p=0.015). Changes in OPG mRNA correlated with changes in BMD (r=-0.53, p=0.027) and CTX (r=0.7, p=0.0044). Down-regulation in RANKL, OPG, RANK mRNA and reduction in bone resorption was also seen in vitro. These results suggest that the expression of RANKL/OPG/RANK in PBMCs are responsive to the slowing in bone turnover/remodeling associated with treatment with estrogen or raloxifene. Further confirmatory studies are needed.
Assuntos
Proteínas de Transporte/genética , Estrogênios/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Glicoproteínas/genética , Leucócitos Mononucleares/metabolismo , Glicoproteínas de Membrana/genética , Cloridrato de Raloxifeno/farmacologia , Receptores Citoplasmáticos e Nucleares/genética , Receptores do Fator de Necrose Tumoral/genética , Idoso , Densidade Óssea/efeitos dos fármacos , Reabsorção Óssea/tratamento farmacológico , Colágeno/sangue , Colágeno Tipo I , Feminino , Glicoproteínas/sangue , Terapia de Reposição Hormonal , Humanos , Estudos Longitudinais , Pessoa de Meia-Idade , Osteoporose/tratamento farmacológico , Osteoprotegerina , Peptídeos/sangue , Pós-Menopausa , Ligante RANK , RNA Mensageiro/sangue , Receptor Ativador de Fator Nuclear kappa-B , Receptores Citoplasmáticos e Nucleares/sangue , Receptores do Fator de Necrose Tumoral/sangue , Moduladores Seletivos de Receptor EstrogênicoRESUMO
Pasteurella multocida toxin (PMT) is an unusual toxin that acts as a mitogen by stimulating various intracellular signalling cascades. Pathways downstream of the G-protein Gq and also downstream of the Rho proteins are activated. Thus PMT action stimulates phospholipase C leading to activation of protein kinase C, an increase in inositol phosphates, and a rise in intracellular calcium. Rho activation of the Rho kinase leads to cytoskeletal reorganisation, tyrosine phosphorylation of the focal adhesion kinase, and activation of the Src proto-oncogene. In addition, signalling through the Ras-MAP kinase signalling pathway is also initiated. PMT is an intracellularly acting toxin, and functional domains that carry out different aspects of its function have been described. The intracellular target of the toxin is currently not known. PMT also acts to inhibit differentiation, in particular of bone cells, where it prevents the formation of mineralised bone nodules in vitro. The toxin is the causative agent of a porcine disease that is characterised by bone resorption. Injection of very low doses of toxin leads to proliferative effects, but at higher doses is lethal. The possible effect of PMT-induced perturbation of signal transduction pathways is discussed.
Assuntos
Proteínas de Bactérias , Toxinas Bacterianas/farmacologia , Diferenciação Celular , Divisão Celular , Osteoblastos/citologia , Pasteurella multocida/patogenicidade , Transdução de Sinais , Células 3T3 , Animais , Proteínas de Ligação ao GTP/metabolismo , Humanos , Camundongos , Mitógenos , Infecções por Pasteurella/microbiologia , Infecções por Pasteurella/fisiopatologia , Infecções por Pasteurella/veterinária , Proto-Oncogene Mas , Suínos , Doenças dos Suínos/microbiologia , Doenças dos Suínos/fisiopatologiaRESUMO
The interstitial collagenases have been suggested to play a critical role in bone formation, remodeling, and cancerogenesis. We have previously shown that during mouse development expression of collagenase-3 (MMP-13) is restricted to bone and cartilage (Gack et al., 1995; Tuckermann et al., 2000) and is affected in mice with altered c-Fos and Cbfa-1 expression (Gack et al., 1994; Porte et al., 1999). In this study, using immunohistochemistry (IHC) and in situ hybridization (ISH) techniques, we have identified cells of the osteoblastic lineage to be the origin of strongly enhanced levels of MMP-13 transcripts in c-fos-induced osteosarcomas. Expression in these cells is further increased in c-fos/c-jun double transgenic mice and paralleled by Cbfa-1 expression. Similarly, in spontaneous and radiation-induced osteosarcomas, both c-Fos and MMP-13 proteins are detectable, suggesting that overexpression of both genes is a characteristic feature of osteosarcomas of different origin. We also observed high levels of MMP-13 in c-Fos-induced chondrosarcomas. In osteoblast-like cells and in cells of late chondrocyte differentiation such as hypertrophic chondrocytes, high levels of MMP-13 transcripts were found. In contrast, in anaplastic areas of the tumors representing highly proliferating chondrocytes, no MMP-13 expression is detectable, suggesting that in addition to Fos/AP-1, bone-specific transcription factors are responsible for restricted expression of collagenase-3/MMP-13 in a specific subset of cells of bone and cartilage in physiology and pathology.