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1.
Electrophoresis ; 39(7): 998-1005, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29330871

RESUMO

In the first part of this publication, the results from an international study evaluating the precision (i.e., repeatability and reproducibility) of N-glycosylation analysis using capillary electrophoresis of APTS-labeled N-glycans were presented. The corresponding results from ultra-high performance liquid chromatography (UHPLC) with fluorescence detection are presented here from 12 participating sites. All participants used the same lot of samples, reagents, and columns to perform the assays. Elution time, peak area and peak area percent values were determined for all peaks ≥0.1% peak area, and statistical analysis was performed following ISO 5725-2 guideline principles. The results demonstrated adequate reproducibility, within any given site as well across all sites, indicating that standard UHPLC-based N-glycan analysis platforms are appropriate for general use.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Corantes Fluorescentes/química , Polissacarídeos/análise , Benzamidas/química , Sítios de Ligação , Eletroforese Capilar/métodos , Glicosilação , Humanos , Limite de Detecção , Reprodutibilidade dos Testes , Espectrometria de Fluorescência/métodos
2.
J Bone Miner Res ; 28(11): 2357-67, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23649605

RESUMO

Bone metastasis, the leading cause of breast cancer-related deaths, is characterized by bone degradation due to increased osteoclastic activity. In contrast, mechanical stimulation in healthy individuals upregulates osteoblastic activity, leading to new bone formation. However, the effect of mechanical loading on the development and progression of metastatic breast cancer in bone remains unclear. Here, we developed a new in vivo model to investigate the role of skeletal mechanical stimuli on the development and osteolytic capability of secondary breast tumors. Specifically, we applied compressive loading to the tibia following intratibial injection of metastatic breast cancer cells (MDA-MB231) into the proximal compartment of female immunocompromised (SCID) mice. In the absence of loading, tibiae developed histologically-detectable tumors with associated osteolysis and excessive degradation of the proximal bone tissue. In contrast, mechanical loading dramatically reduced osteolysis and tumor formation and increased tibial cancellous mass due to trabecular thickening. These loading effects were similar to the baseline response we observed in non-injected SCID mice. In vitro mechanical loading of MDA-MB231 in a pathologically relevant 3D culture model suggested that the observed effects were not due to loading-induced tumor cell death, but rather mediated via decreased expression of genes interfering with bone homeostasis. Collectively, our results suggest that mechanical loading inhibits the growth and osteolytic capability of secondary breast tumors after their homing to the bone, which may inform future treatment of breast cancer patients with advanced disease.


Assuntos
Neoplasias Ósseas/fisiopatologia , Neoplasias da Mama/patologia , Neoplasias da Mama/fisiopatologia , Osteólise/patologia , Osteólise/fisiopatologia , Tíbia/fisiopatologia , Animais , Neoplasias Ósseas/complicações , Neoplasias Ósseas/diagnóstico por imagem , Neoplasias Ósseas/secundário , Reabsorção Óssea/complicações , Reabsorção Óssea/diagnóstico por imagem , Reabsorção Óssea/patologia , Reabsorção Óssea/fisiopatologia , Neoplasias da Mama/complicações , Neoplasias da Mama/diagnóstico por imagem , Linhagem Celular Tumoral , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Camundongos SCID , Osteólise/complicações , Osteólise/diagnóstico por imagem , Tíbia/patologia , Alicerces Teciduais/química , Suporte de Carga , Microtomografia por Raio-X
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