Investigating c-Met Signaling in Non-Small Cell Lung Cancer

Despite improvements in preventive, diagnostic, and therapeutic approaches, lung cancer remains the leading cause of cancer-related deaths in the U.S. There are currently no effective therapies for those diagnosed with later stage lung cancer. Recent efforts have focused on targeting specific lung cancer-related growth pathways, such as the hepatocyte growth factor (HGF)/c-Met signaling pathway. HGF/c-Met signaling plays a critical role in mediating proliferation, angiogenesis, invasion, and inflammatory responses in non-small cell lung cancer (NSCLC). This signaling pathway also confers resistance to therapies targeting other growth factor pathways such as epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF). This study focuses on two aspects of HGF/c-Met signaling relevant to lung cancer: HGF stimulated c-Met angiogenic response and EGFR ligand-induced c-Met pro-cancer signaling. We previously reported airway expression of a human HGF transgene (HGF TG) produced mice that were more susceptible to lung tumorigenesis induced by a tobacco carcinogen. Here we show untreated HGF TG mice display enhanced vascularization and lymph vessel formation in the lungs compared to wild-type (WT) littermates, as ascertained by microvessel density. Whole lung microarray analysis consistently found significant decreases in expression of genes involved in angiogenesis including the VEGF family of genes (Vegfa,Vegfb, Vegfc, Vegfd/Figf) at 10, 20, and 40 weeks of age in HGF TG animals compared to WT littermates. HGF TG lung tumors derived from carcinogen treated HGF TG mice demonstrated reduction in VEGF genes with an increased expression of five Cxcl family genes including Cxcl1 and Cxcl2 (murine forms of IL-8). Thus, increased vascularization produced by airway over-expression of HGF may occur through direct activation of c-Met on endothelial cells and expression of inflammatory mediators, rather than induction of VEGF pathways. Ligand-independent delayed and prolonged activation of c-Met has also been demonstrated previously by our laboratory via cross-talk from the EGFR pathway. Here we show that prolonged activation of STAT3 by EGFR-ligands is dependent on delayed c-Met activation. Inhibition of c-Met, by PHA665752, eliminates EGFR stimulated activation of STAT3 at delayed time points. The failure of Src inhibition, by PP2, to block delayed phospho-STAT3, and the co-immunoprecipitation of STAT3 with c-Met 8-24hours post EGF stimulation, suggests STAT3 is directly activated by c-Met. These data reinforce the idea that delayed c-Met activation is utilized by EGFR to potentiate its full biological effects through STAT3. Both ligand-dependent transient and ligand-independent delayed c-Met activation appear to be important in lung tumorigenesis. The findings of this study support future development of novel HGF/c-Met and EGFR combination therapies in NSCLC

A Hybrid Decision Model for Improving Warehouse Efficiency in a Process-oriented View

published_fina

Laser Hair Removal: Comparative Study of Light Wavelength and its Effect on Laser Hair Removal

For some people, hirsuteness (having excess body hair) can be an embarrassing problem. Many attempts have been made to find a solution to these problems, including electrolysis, tweezing, shaving, and waxing. However, most of these solutions are painful, are not useful in treating large areas of skin, or are not permanent. Laser hair removal stands out amongst these other methods as a permanent method of reducing hirsuteness that can cover large areas of the body, such as the chest or the legs. While laser hair removal is a widely used technology, few studies have explored the physical aspects of why it works so well. More specifically, there is a significant lack of computer models that show how temperature profiles look inside the hair and surrounding skin. Using the physical properties and dimensions of hair, we constructed a model of the hair that approximates how actual hair resides in the skin. Using COMSOL Multiphysics, we tested this model with five different lasers of varying wavelengths in order to determine the relationship between laser wavelength and temperature in the hair. Using a laser pulse duration of 0.01 seconds (10 milliseconds), we found a positive correlation between wavelength and temperature, with all wavelengths except the lowest (595 nm) achieving a temperature above the threshold temperature required for hair destruction. In addition, while all lasers caused a temperature rise in the surrounding skin, the extent of thermal damage was minimal. However, since we could not find physical properties of the hair follicle itself, we were forced to approximate those properties using the hair shaft properties, ultimately leading us to treat the follicle and shaft as one entity. This is a slight limitation with our design. Regardless, we have provided a greater understanding of the physiological temperatures involved in hair removal, and have reinforced the fact that laser hair removal can be a safe method and effective method for treating hirsuteness by showing that hair follicles can be heated to a temperature that kill them by using lasers, and that this heating does not severely or irreparably damage surrounding skin

Analysis of mixtures using next generation sequencing of mitochondrial DNA hypervariable regions

Aim To apply massively parallel and clonal sequencing (next generation sequencing or NGS) to the analysis of forensic mixed samples. Methods A duplex polymerase chain reaction (PCR) assay targeting the mitochondrial DNA (mtDNA) hypervariable regions I/II (HVI/HVII) was developed for NGS analysis on the Roche 454 GS Junior instrument. Eight sets of multiplex identifier-tagged 454 fusion primers were used in a combinatorial approach for amplification and deep sequencing of up to 64 samples in parallel. Results This assay was shown to be highly sensitive for sequencing limited DNA amounts ( ~ 100 mtDNA copies) and analyzing contrived and biological mixtures with low level variants ( ~ 1%) as well as “complex” mixtures (≥3 contributors). PCR artifact “hybrid” sequences generated by jumping PCR or template switching were observed at a low level (<2%) in the analysis of mixed samples but could be eliminated by reducing the PCR cycle number. Conclusion This study demonstrates the power of NGS technologies targeting the mtDNA HVI/HVII regions for analysis of challenging forensic samples, such as mixtures and specimens with limited DNA

Mapping the Overall Individual Visitors’ Experiences at Sarawak Cultural Village as a Living Museum Site

A living museums or non-living museums were an integral tourist destination as it was important to understand visitors’ experiences during the visitation. This study aimed to explore the overall visitor experiences at Sarawak Cultural Village (SCV) at an individual level. The visitors’ experience was seen vital as it will also help in improving SCV in the future. The qualitative research method was used for this study using interview and photovoice. The findings from interview sessions and the photo voices taken during the visits were analysed using thematic analysis to produce the visitors’ experience. A total of 20 respondents participated in this study and were randomly selected during the two-weeks of data collection. The result of the study showed that SCV was a unique and authentic place to visit when comes to a heritage and cultural site. An initial result suggested that there are 18 sub-themed emerged from this study such as memory, learning, emotion, engagement, knowledge discovery, interaction, natural environment, aesthetic, fun and entertainment, involvement, uniqueness of culture, rare object, creativity, originality, culture exposure, antique artefacts, unique architecture, and costume. A further analysis showed that these 18 sub-themes can be categorised into 6 main themes such as: memory experience, learning experience, emotional experience, aesthetic experience, interactive experience and cultural experience. Further study should be done to explore the use of social media and other methods of data collection such as journey map

Combining the Multitargeted Tyrosine Kinase Inhibitor Vandetanib with the Antiestrogen Fulvestrant Enhances Its Antitumor Effect in Non-small Cell Lung Cancer

IntroductionEstrogen is known to promote proliferation and to activate the epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC). Vascular endothelial growth factor (VEGF) is a known estrogen responsive gene in breast cancer. We sought to determine whether the VEGF pathway is also regulated by estrogen in lung cancer cells, and whether combining an inhibitor of the ER pathway with a dual vascular endothelial growth factor receptor (VEGFR)/EGFR inhibitor would show enhanced antitumor effects.MethodsWe examined activation of EGFR and expression of VEGF in response to β-estradiol, and the antitumor activity of the multitargeted VEGFR/EGFR/RET inhibitor, vandetanib, when combined with the antiestrogen fulvestrant both in vitro and in vivo.ResultsNSCLC cells expressed VEGFR-3 and EGFR. Vandetanib treatment of NSCLC cells resulted in inhibition of EGFR and VEGFR-3 and inhibition of β-estradiol-induced P-MAPK activation, demonstrating that vandetanib blocks β-estradiol-induced EGFR signaling. Treatment with β-estradiol stimulated VEGFA mRNA and protein (p < 0.0001 over baseline), suggesting estrogenic signaling causes heightened VEGFA pathway activation. This estrogenic induction of VEGFA mRNA seems largely dependent on cross-talk with EGFR. Long-term vandetanib treatment also significantly increased ERβ protein expression. The combination of vandetanib with fulvestrant maximally inhibited cell growth compared with single agents (p < 0.0001) and decreased tumor xenograft volume by 64%, compared with 51% for vandetanib (p < 0.05) and 23% for fulvestrant (p < 0.005). Antitumor effects of combination therapy were accompanied by a significant increase in apoptotic cells compared with single agents.ConclusionsFulvestrant may enhance effects of vandetanib in NSCLC by blocking estrogen-driven activation of the EGFR pathway

Exploring the relationship between sexual compulsivity and attentional bias to sex-related words in a cohort of sexually active individuals

Background/Aims: If sexual compulsivity and other addictive behaviours share common aetiology, contemporary proposals about the role of attentional processes in understanding addictive behaviours are relevant. Methods: To examine attentional biases for sex-related words amongst sexually active individuals and the relationship between sexual compulsivity and sexual behavioural engagement with attentional bias, 55 sexually active individuals completed a modified Stroop task and the sexual compulsivity scale. Results: Findings showed attentional bias towards sex-related stimuli among sexually active participants. In addition, amongst those with low levels of sexual compulsivity, levels of attentional bias were the same across all levels of sexual experience. Amongst those with higher levels of sexual compulsivity, greater attentional bias was linked with lower levels of sexual experience. Conclusion: Attentional preference for concern-related stimuli varies as a function of the interaction between how long a person has been active sexually and how compulsive their sexual behaviour is

The degree of microbiome complexity influences the epithelial response to infection

<p>Abstract</p> <p>Background</p> <p>The human microflora is known to be extremely complex, yet most pathogenesis research is conducted in mono-species models of infection. Consequently, it remains unclear whether the level of complexity of a host's indigenous flora can affect the virulence potential of pathogenic species. Furthermore, it remains unclear whether the colonization by commensal species affects a host cell's response to pathogenic species beyond the direct physical saturation of surface receptors, the sequestration of nutrients, the modulation of the physico-chemical environment in the oral cavity, or the production of bacteriocins. Using oral epithelial cells as a model, we hypothesized that the virulence of pathogenic species may vary depending on the complexity of the flora that interacts with host cells.</p> <p>Results</p> <p>This is the first report that determines the global epithelial transcriptional response to co-culture with defined complex microbiota. In our model, human immortalized gingival keratinocytes (HIGK) were infected with mono- and mixed cultures of commensal and pathogenic species. The global transcriptional response of infected cells was validated and confirmed phenotypically. In our model, commensal species were able to modulate the expression of host genes with a broad diversity of physiological functions and antagonize the effect of pathogenic species at the cellular level. Unexpectedly, the inhibitory effect of commensal species was <it>not </it>correlated with its ability to inhibit adhesion or invasion by pathogenic species.</p> <p>Conclusion</p> <p>Studying the global transcriptome of epithelial cells to single and complex microbial challenges offers clues towards a better understanding of how bacteria-bacteria interactions and bacteria-host interactions impact the overall host response. This work provides evidence that the degree of complexity of a mixed microbiota <it>does </it>influence the transcriptional response to infection of host epithelial cells, and challenges the current dogma regarding the <it>potential </it>versus the <it>actual </it>pathogenicity of bacterial species. These findings support the concept that members of the commensal oral flora have evolved cellular mechanisms that directly modulate the host cell's response to pathogenic species and dampen their relative pathogenicity.</p