Breaking up Prolonged Sitting does not Alter Postprandial Glycemia in Young, Normal-Weight Men and Women.

A randomized, controlled, cross-over study was used to investigate the effects of breaking up prolonged sitting with low intensity physical activity on postprandial blood glucose concentrations in healthy, young, normal-weight adults. 14 men (n=6) and women (n=8) were assigned to 2.5 h of prolonged sitting (CON) and 2.5 h of prolonged sitting with 2-min bouts of walking every 20 min (LIPA). After ingesting a standardized test drink, capillary blood was sampled every 10 min to establish a postprandial blood glucose profile. Based on individual glucose responses, peak blood glucose, time-to-peak glucose, and incremental area under the glucose curve (iAUC) were determined. Paired sample t-tests were used to detect differences between trials. Peak blood glucose (p=0.55) and iAUC (CON: 252 mmol·L-1·2.5 h-1 [163-340]; LIPA: 214 mmol·L-1·2.5 h-1 [146-282]; p=0.45) were not different between trials. Also, time-to-peak glucose was not different between LIPA and CON (p=0.37). Taking advantage of high temporal resolution blood glucose profiles, we showed that breaking up prolonged sitting with low-intensity physical activity does not alter the postprandial blood glucose response in young, healthy, normal-weight adults. Our results indicate that postprandial glycemic control is maintained during prolonged sitting in young, healthy adults.

MCF-10A-NeoST: a new cell system for studying cell-ECM and cell-cell interactions in breast cancer.

PURPOSE: There is a continuing need for genetically matched cell systems to model cellular behaviors that are frequently observed in aggressive breast cancers. EXPERIMENTAL DESIGN: We report here the isolation and initial characterization of a spontaneously arising variant of MCF-10A cells, NeoST, which provides a new model to study cell adhesion and signal transduction in breast cancer. RESULTS: NeoST cells recapitulate important biological and biochemical features of metastatic breast cancer, including anchorage-independent growth, invasiveness in three-dimensional reconstituted membranes, loss of E-cadherin expression, and increased tyrosine kinase activity. A comprehensive analysis of tyrosine kinase expression revealed overexpression or functional activation of the Axl, FAK, and EphA2 tyrosine kinases in transformed MCF-10A cells. CONCLUSIONS: MCF-10A and these new derivatives provide a genetically matched model to study defects in cell adhesion and signaling that are relevant to cellular behaviors that often typify aggressive breast cancer cells.

Hsp27-induced MMP-9 expression is influenced by the Src tyrosine protein kinase yes.

The small heat shock protein hsp27 is associated with aggressive tumor behavior in certain subsets of breast cancer patients. Previously we demonstrated that hsp27 overexpression in breast cancer cells increased in vitro and in vivo invasiveness, suggesting that hsp27 influences the metastatic process. To investigate this role for hsp27, we have utilized MDA-MB-231 breast cancer cells that overexpress hsp27 and cDNA expression array technology. We demonstrate that hsp27 overexpression up-regulates MMP-9 expression and activity and down-regulates Yes expression. Furthermore, our results suggest that Yes may be involved in regulating MMP-9 expression, as well as in vitro invasion. Reconstitution of Yes expression by transfection into hsp27-overexpressing cells decreased MMP-9 expression, and increased in vitro invasiveness, abrogating the phenotype conferred by hsp27 overexpression. Therefore, our results provide a new potential mechanism by which hsp27 affects the metastatic cascade-through regulation of MMP-9 and Yes expression.

Tissue architecture and breast cancer: the role of extracellular matrix and steroid hormones.

The changes in tissue architecture that accompany the development of breast cancer have been the focus of investigations aimed at developing new cancer therapeutics. As we learn more about the normal mammary gland, we have begun to understand the complex signaling pathways underlying the dramatic shifts in the structure and function of breast tissue. Integrin-, growth factor-, and steroid hormone-signaling pathways all play an important part in maintaining tissue architecture; disruption of the delicate balance of signaling results in dramatic changes in the way cells interact with each other and with the extracellular matrix, leading to breast cancer. The extracellular matrix itself plays a central role in coordinating these signaling processes. In this review, we consider the interrelationships between the extracellular matrix, integrins, growth factors, and steroid hormones in mammary gland development and function.

Hsp27 overexpression inhibits doxorubicin-induced apoptosis in human breast cancer cells.

Previously we demonstrated that heat shock protein 27 (hsp27) overexpression confers resistance to the chemotherapeutic agent doxorubicin in MDA-MB-231 breast cancer cells. Since induction of apoptosis is one underlying mechanism of chemotherapeutic drug action, we investigated the effect of hsp27 overexpression on doxorubicin-induced apoptosis, finding that hsp27 protects MDA-MB-231 cells from apoptosis. We also examined expression of the doxorubicin target, topoisomerase II (topo II), in control and hsp27-overexpressing stable transfectants, as topo II expression is important for both drug sensitivity and the initiation of apoptosis by doxorubicin. The relative levels of both topo IIalpha and beta were higher in the controls than the hsp27-overexpressing clones, suggesting that the apoptotic protective effect of hsp27 overexpression in MDA-MB-231 cells is associated with altered topo II expression.

Statistical analysis of array expression data as applied to the problem of tamoxifen resistance.

BACKGROUND: Although the emerging complementary DNA (cDNA) array technology holds great promise to discern complex patterns of gene expression, its novelty means that there are no well-established standards to guide analysis and interpretation of the data that it produces. We have used preliminary data generated with the CLONTECH Atlas human cDNA array to develop a practical approach to the statistical analysis of these data by studying changes in gene expression during the development of acquired tamoxifen resistance in breast cancer. METHODS: For hybridization to the array, we prepared RNA from MCF-7 human breast cell tumors, isolated from our athymic nude mouse xenograft model of acquired tamoxifen resistance during estrogen-stimulated, tamoxifen-sensitive, and tamoxifen-resistant growth. Principal components analysis was used to identify genes with altered expression. RESULTS AND CONCLUSIONS: Principal components analysis yielded three principal components that are interpreted as 1) the average level of gene expression, 2) the difference between estrogen-stimulated gene expression and the average of tamoxifen-sensitive and tamoxifen-resistant gene expression, and 3) the difference between tamoxifen-sensitive and tamoxifen-resistant gene expression. A bivariate (second and third principal components) 99% prediction region was used to identify outlier genes that exhibit altered expression. Two representative outlier genes, erk-2 and HSF-1 (heat shock transcription factor-1), were chosen for confirmatory study, and their predicted relative expression levels were confirmed in western blot analysis, suggesting that semiquantitative estimates are possible with array technology. IMPLICATIONS: Principal components analysis provides a useful and practical method to analyze gene expression data from a cDNA array. The method can identify broad patterns of expression alteration and, based on a small simulation study, will likely provide reasonable power to detect moderate-sized alterations in clinically relevant genes.

Quercetin inhibits heat shock protein induction but not heat shock factor DNA-binding in human breast carcinoma cells.

The flavonoid quercetin inhibits the heat-induced synthesis of heat shock proteins (hsps) in a variety of cell lines. To determine whether quercetin could inhibit hsp expression in breast cancer cells, we used the human breast cancer cell line, MDA-MB-231. Treatment of these cells with quercetin decreased the heat-induced synthesis of hsp27 and hsp70. However, inhibition of hsp expression did not correspond with the reduced ability of heat shock transcription factors (HSFs) to bind DNA. Furthermore, while quercetin treatment inhibited HSF2 expression, it only slightly affected HSF1 expression in breast cancer cells. In contrast, quercetin inhibited both HSF DNA-binding activity and HSF expression in HeLa cells. Our studies suggest that quercetin's action is cell-type specific, and in breast cancer cells may involve regulation of HSF transcriptional activity, rather than regulation of its DNA-binding activity.

Multiple negative and positive cis-acting elements control the expression of the murine CD4 gene.

The cis-acting elements located within 15 kb 5' of the murine CD4 gene transcriptional start site and the first intron of the CD4 gene have been investigated using deletion constructs. Our transient transfection data indicate that the expression of the murine CD4 gene is controlled by multiple positive and negative regulatory cis-acting elements. There are at least two cis-acting elements that have a positive effect on the expression of the CD4 gene and at least four regions of DNA that have a negative effect. The positive control elements are located about 13.5 kb 5' of the promoter and within the flanking sequences of the first intron. The DNA between the 5' enhancer and the promoter contains at least two regions that exert a negative effect on CD4 expression. In addition to the positive effect that the first intron has on CD4 expression, there are two regions within the first intron that have a negative effect. These two negative regulatory elements correspond to two T-cell-specific DNase I hypersensitive sites found in the first intron.

A comparison between two methods of measuring pressure in the pharyngeal airway: transducer probe versus open catheter.

A new multi-transducer probe system for measuring pharyngeal pressures was compared with an established open catheter system. Pharyngeal pressure measurements were made at the same time, and site, in subjects awake, at unmodified and with artificially increased nasal airway resistances, and during sleep documented by polysomnography. The two systems yielded almost identical results. It is anticipated that the multi-transducer probe system will prove of clinical value.