Tyrosine-kinase inhibition results in EGFR clustering at focal adhesions and consequent exocytosis in uPAR down-regulated cells of Head and Neck cancers

<p>Abstract</p> <p>Background</p> <p>Antisense (AS) induced down-regulation of uPAR in ACCS adenoid-cyctic carcinoma cells decreased the cellular adhesion and invasion on various extracellular matrices. Additionally, ACCS-AS cells showed an increased EGFR expression and other behavioral similarities to NA-SCC, a typical highly proliferative but less invasive squamous cell carcinoma (SCC) cell line of the head and neck. ACCS, ACCS-AS and NA-SCC cells were used to elucidate the relationships between uPAR down-regulation and EGFR inhibition.</p> <p>Results</p> <p>Tyrosine kinase inhibitor Gefitinib (IRESSA, ZD 1839) significantly reduced the chemotactic cell migration and adhesion. This was associated with reduced EGFR and ERK activation. In addition, anti-proliferative effect of gefitinib in uPAR down-regulated ACCS-AS was significantly higher than parental ACCS, to levels comparable to gefitinib-sensitive NA-SCC cells. This was evidenced by both reduced dosage and duration of treatment. Furthermore, time-lapse videography showed that treatment with gefitinib was also associated with cell rounding and loss of pseudopodia, mostly in ACCS-AS rather than parental ACCS cells. There were also evidences of formation and exocytosis of vacuole-like structures in ACCS-AS, as well as NA-SCC, but not in parental ACCS cells. Interestingly, immunocytochemistry showed that the exocytotic vacuoles actually contained de-activated EGFR.</p> <p>Conclusion</p> <p>Our results suggested that down-regulation of uPAR affected the fate of EGFR in high EGFR expressing cells. Furthermore, combining the uPAR down-regulation with EGFR inhibition showed a synergistic anti-tumor effect and might provide an alternative method to increase anti-proliferative effect of tyrosine kinase inhibitors with lower doses and duration to reduce their side effects during cancer control.</p

Impaired germ cell development due to compromised cell cycle progression in Skp2-deficient mice

BACKGROUND: The gonads are responsible for the production of germ cells through both mitosis and meiosis. Skp2 is the receptor subunit of an SCF-type ubiquitin ligase and is a major regulator of the progression of cells into S phase of the cell cycle, which it promotes by mediating the ubiquitin-dependent degradation of p27, an inhibitor of cell proliferation. However, the role of the Skp2-p27 pathway in germ cell development remains elusive. RESULTS: We now show that disruption of Skp2 in mice results in a marked impairment in the fertility of males, with the phenotypes resembling Sertoli cell-only syndrome in men. Testes of Skp2(-/- )mice manifested pronounced germ cell hypoplasia accompanied by massive apoptosis in spermatogenic cells. Flow cytometry revealed an increased prevalence of polyploidy in spermatozoa, suggesting that the aneuploidy of these cells is responsible for the induction of apoptosis. Disruption of the p27 gene of Skp2(-/- )mice restored germ cell development, indicating that the testicular hypoplasia of Skp2(-/- )animals is attributable to the antiproliferative effect of p27 accumulation. CONCLUSION: Our results thus suggest that compromised cell cycle progression caused by the accumulation of p27 results in aneuploidy and the induction of apoptosis in gonadal cells of Skp2(-/- )mice. The consequent reduction in the number of mature gametes accounts for the decreased fertility of these animals. These findings reinforce the importance of the Skp2-p27 pathway in cell cycle regulation and in germ cell development

THE EFFECTS OF MICRO- AND MACRO-SCALE GEOMETRIC PARAMETERS ON PERFORMANCE OF THE PLEATED AEROSOL FILTERS

While most filters are made of pleated fibrous media, almost all existing theories of aerosol filtration are developed for flat media placed perpendicular to the air flow. Expressions developed for flat sheet media do not provide accurate information directly useful for designing a pleated filter, and therefore, most progress made in developing pleated filters is based on empiricism. This study is aimed at establishing an enabling knowledge that allows for a better design and optimization of pleated aerosol filters. This study is focused on developing a predictive simulation method that accounts for the influence of a filter’s micro-scale geometric parameters, such as fiber orientation, as well as its macro-scale features, like pleat shape, in predicting the transient pressure drop and collection efficiency with or without the effects of dust loading. The dual-scale simulation method developed in this work is believed to be the only feasible approach for design and optimization of pleated aerosol filters with the current academic-level computational power. Our study is divided into two major tasks of micro- and macro-scale modeling. Our micro-scale studies are comprised of a series of CFD simulations conducted in virtual 2-D or 3-D fibrous geometries that resemble the internal micro-structure of a fibrous medium. These simulations are intended to isolate the effects of each micro structural parameter and study its influence on the performance of the filter medium. In detail, it is intended to propose a method to predict the performance of micro-structures with fiber size distribution. Also, the effects of micro-structural fiber orientation were investigated. Moreover, we offered methodology to predict the performance of noncircular fibers using available analytical expressions for circular fibers. It is shown that the circumscribed circle for a trilobal shaped fiber gives the best prediction for collection efficiency. In macro-scale simulations, on the other hand, the filter medium is treated as a lumped porous material with its properties obtained via micro-scale simulations. Our results showed that more number of pleats helps better performance of pleated filters, however, if the pleat channel becomes blocked by dust cake then this effect is no longer valid

The Expression and Localization of N-Myc Downstream-Regulated Gene 1 in Human Trophoblasts

The protein N-Myc downstream-regulated gene 1 (NDRG1) is implicated in the regulation of cell proliferation, differentiation, and cellular stress response. NDRG1 is expressed in primary human trophoblasts, where it promotes cell viability and resistance to hypoxic injury. The mechanism of action of NDRG1 remains unknown. To gain further insight into the intracellular action of NDRG1, we analyzed the expression pattern and cellular localization of endogenous NDRG1 and transfected Myc-tagged NDRG1 in human trophoblasts exposed to diverse injuries. In standard conditions, NDRG1 was diffusely expressed in the cytoplasm at a low level. Hypoxia or the hypoxia mimetic cobalt chloride, but not serum deprivation, ultraviolet (UV) light, or ionizing radiation, induced the expression of NDRG1 in human trophoblasts and the redistribution of NDRG1 into the nucleus and cytoplasmic membranes associated with the endoplasmic reticulum (ER) and microtubules. Mutation of the phosphopantetheine attachment site (PPAS) within NDRG1 abrogated this pattern of redistribution. Our results shed new light on the impact of cell injury on NDRG1 expression patterns, and suggest that the PPAS domain plays a key role in NDRG1's subcellular distribution. © 2013 Shi et al

c-MET Protects Breast Cancer Cells from Apoptosis Induced by Sodium Butyrate

Sodium Butyrate (NaBu) is regarded as a potential reagent for cancer therapy. In this study, a specific breast cancer cell population that is resistant NaBu treatment was identified. These cells possess cancer stem cell characters, such as the capability of sphere formation in vitro and high tumor incident rate (85%) in mouse model. Forty percent of the NaBu resistant cells express the cancer stem cells marker, the CD133, whereas only 10% intact cells present the CD133 antigen. Furthermore, the endogenous expressing c-MET contributes to the survival of cancer stem cell population from the treatment of NaBu. The CD133+ group also presents a higher level of c-MET. A combination treatment of MET siRNA and NaBu efficiently prohibited the breast cancer progression, and the incident rate of the tumor decrease to 18%. This study may help to develop a new and alternative strategy for breast cancer therapy

Effect of combined treatment with alendronate and calcitriol on femoral neck strength in osteopenic rats

<p>Abstract</p> <p>Background</p> <p>Hip fracture is associated with pronounced morbidity and excess mortality in elderly women with postmenopausal osteoporosis. Many drugs have been developed to treat osteoporosis and to reduce the risk of osteoporotic fractures. We investigated the effects of combined alendronate and vitamin D₃treatment on bone mass and fracture load at the femoral neck in ovariectomized (OVX) rats, and evaluated the relationship between bone mass parameters and femoral neck strength.</p> <p>Methods</p> <p>Thirty 12-week-old female rats underwent either a sham-operation (n = 6) or OVX (n = 24). Twenty weeks later, OVX rats were further divided into four groups and received daily doses of either saline alone, 0.1 mg/kg alendronate, 0.1 μg/kg calcitriol, or a combination of both two drugs by continuous infusion via Alzet mini-osmotic pumps. The sham-control group received saline alone. After 12 weeks of treatment, femoral necks were examined using peripheral quantitative computed tomography (pQCT) densitometry and mechanical testing.</p> <p>Results</p> <p>Saline-treated OVX rats showed significant decreases in total bone mineral content (BMC) (by 28.1%), total bone mineral density (BMD) (by 9.5%), cortical BMC (by 26.3%), cancellous BMC (by 66.3%), cancellous BMD (by 29.0%) and total cross-sectional bone area (by 30.4%) compared with the sham-control group. The combined alendronate and calcitriol treatments improved bone loss owing to estrogen deficiency. On mechanical testing, although OVX significantly reduced bone strength of the femoral neck (by 29.3%) compared with the sham-control group, only the combined treatment significantly improved the fracture load at the femoral neck in OVX rats to the level of the sham-controls. The correlation of total BMC to fracture load was significant, but that of total BMD was not.</p> <p>Conclusion</p> <p>Our results showed that the combined treatment with alendronate and calcitriol significantly improved bone fragility of the femoral neck in OVX osteopenic rats.</p

Recapitulation of tumor heterogeneity and molecular signatures in a 3D brain cancer model with decreased sensitivity to histone deacetylase inhibition

INTRODUCTION Physiologically relevant pre-clinical ex vivo models recapitulating CNS tumor micro-environmental complexity will aid development of biologically-targeted agents. We present comprehensive characterization of tumor aggregates generated using the 3D Rotary Cell Culture System (RCCS). METHODS CNS cancer cell lines were grown in conventional 2D cultures and the RCCS and comparison with a cohort of 53 pediatric high grade gliomas conducted by genome wide gene expression and microRNA arrays, coupled with immunohistochemistry, ex vivo magnetic resonance spectroscopy and drug sensitivity evaluation using the histone deacetylase inhibitor, Vorinostat. RESULTS Macroscopic RCCS aggregates recapitulated the heterogeneous morphology of brain tumors with a distinct proliferating rim, necrotic core and oxygen tension gradient. Gene expression and microRNA analyses revealed significant differences with 3D expression intermediate to 2D cultures and primary brain tumors. Metabolic profiling revealed differential profiles, with an increase in tumor specific metabolites in 3D. To evaluate the potential of the RCCS as a drug testing tool, we determined the efficacy of Vorinostat against aggregates of U87 and KNS42 glioblastoma cells. Both lines demonstrated markedly reduced sensitivity when assaying in 3D culture conditions compared to classical 2D drug screen approaches. CONCLUSIONS Our comprehensive characterization demonstrates that 3D RCCS culture of high grade brain tumor cells has profound effects on the genetic, epigenetic and metabolic profiles of cultured cells, with these cells residing as an intermediate phenotype between that of 2D cultures and primary tumors. There is a discrepancy between 2D culture and tumor molecular profiles, and RCCS partially re-capitulates tissue specific features, allowing drug testing in a more relevant ex vivo system

Prediction of Elevated Temperature Fatigue Crack Growth Rates in TI-6AL-4V Alloy – Neural Network Approach

The results obtained from two experimental test programs (TP-1 and TP-2) were used to train neural networks to predict elevated temperature, fatigue crack growth rates in Ti-6Al-4V alloy. Two programs, TP-1 and TP-2, were conducted at room and elevated temperatures under high humidity and laboratory air environments, respectively. While elevated temperature effects were investigated in TP-2, stress ratio effects were studied in TP-1 using several stress ratios. Networks were trained using the elevated temperature data to predict the crack growth rates at a given stress intensity under different temperatures. The experimental and predicted fatigue crack growth rates showed a least squared error of 0.03. Thus, this approach was found to predict fatigue crack growth rates in Ti-6Al-4V alloy at elevated temperatures

Prediction of Elevated Temperature Fatigue Crack Growth Rates in TI-6AL-4V Alloy – Neural Network Approach

The results obtained from two experimental test programs (TP-1 and TP-2) were used to train neural networks to predict elevated temperature, fatigue crack growth rates in Ti-6Al-4V alloy. Two programs, TP-1 and TP-2, were conducted at room and elevated temperatures under high humidity and laboratory air environments, respectively. While elevated temperature effects were investigated in TP-2, stress ratio effects were studied in TP-1 using several stress ratios. Networks were trained using the elevated temperature data to predict the crack growth rates at a given stress intensity under different temperatures. The experimental and predicted fatigue crack growth rates showed a least squared error of 0.03. Thus, this approach was found to predict fatigue crack growth rates in Ti-6Al-4V alloy at elevated temperatures

但馬国 時郡 銭1貫文

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