Sulindac modulates secreted protein expression from LIM1215 colon carcinoma cells prior to apoptosis

Colorectal cancer (CRC) is a major cause of mortality in Western populations. Growing evidence from human and rodent studies indicate that nonsteroidal anti-inflammatory drugs (NSAIDs) cause regression of existing colon tumors and act as effective chemopreventive agents in sporadic colon tumor formation. Although much is known about the action of the NSAID sulindac, especially its role in inducing apoptosis, mechanisms underlying these effects is poorly understood. In previous secretome-based proteomic studies using 2D-DIGE/MS and cytokine arrays we identified over 150 proteins released from the CRC cell line LIM1215 whose expression levels were dysregulated by treatment with 1 mM sulindac over 16 h; many of these proteins are implicated in molecular and cellular functions such as cell proliferation, differentiation, adhesion, angiogenesis and apoptosis (Ji et al., Proteomics Clin. Appl. 2009, 3, 433–451). We have extended these studies and describe here an improved protein/peptide separation strategy that facilitated the identification of 987 proteins and peptides released from LIM1215 cells following 1 mM sulindac treatment for 8 h preceding the onset of apoptosis. This peptidome separation strategy involved fractional centrifugal ultrafiltration of concentrated cell culture media (CM) using nominal molecular weight membrane filters (NMWL 30 K, 3 K and 1 K). Proteins isolated in the > 30 K and 3–30 K fractions were electrophoretically separated by SDS-PAGE and endogenous peptides in the 1–3 K membrane filter were fractioned by RP-HPLC; isolated proteins and peptides were identified by nanoLC-MS–MS. Collectively, our data show that LIM1215 cells treated with 1 mM sulindac for 8 h secrete decreased levels of proteins associated with extracellular matrix remodeling (e.g., collagens, perlecan, syndecans, filamins, dyneins, metalloproteinases and endopeptidases), cell adhesion (e.g., cadherins, integrins, laminins) and mucosal maintenance (e.g., glycoprotein 340 and mucins 5 AC, 6, and 13). A salient finding of this study was the increased proteolysis of cell surface proteins following treatment with sulindac for 8 h (40% higher than from untreated LIM1215 cells); several of these endogenous peptides contained C-terminal amino acids from transmembrane domains indicative of regulated intramembrane proteolysis (RIP). Taken together these results indicate that during the early-stage onset of sulindac-induced apoptosis (evidenced by increased annexin V binding, dephosphorylation of focal adhesion kinase (FAK), and cleavage of caspase-3), 1 mM sulindac treatment of LIM1215 cells results in decreased expression of secreted proteins implicated in ECM remodeling, mucosal maintenance and cell–cell-adhesion. This article is part of a Special Issue entitled: An Updated Secretome.     

Influence of carbon and nitrogen sources on antifungal metabolite production by bacterium associated with entomopathogenic nematode against Penicillium expansum

A specific symbiotic Bacillus sp. isolated from a rhabditid entomopathogenic nematode, Rhabditis (Oscheius) sp. was found to produce large number of bioactive compounds. The present study was conducted to determine the effect of carbon and nitrogen sources for the production of antimicrobial substances by Bacillus sp. The yield of the crude antimicrobial substances and antimicrobial activity against the test micro-organism also differed significantly when carbon and nitrogen sources in the fermentation media were changed. The antifungal activity was significantly high in yeast extract plus fructose (46.5?±?2.12?mm) followed by yeast extract plus maltose, beef extract plus fructose and meat infusion plus glucose. High pressure liquid chromatography analysis of the crude antimicrobial substances revealed different peaks with different retention time indicating that they produced different compounds. When the carbon source was not included in the fermentation media, the antimicrobial production was substantially reduced. The results indicate that carbon source in the fermentation media plays a vital role in the production of antifungal substances. It is concluded that yeast extract and fructose as nitrogen and carbon sources produced maximum activity, which can effectively control the blue mould caused by Penicillium expansum in apples and pears.     

Newspaper Portrayals,Local Policies,and Dog-Supportive Public Space: Who's Wagging Whom?

ABSTRACT

Dog-walking promotes physical activity and positive social interactions, and thus dog-walking has implications for people's physical and mental health. As a result, experts in public health have endorsed designing and managing public space to support dog-walking. Nevertheless, the presence of dogs in public space is subject to negotiation and can be controversial. Generally, municipal governments or local councils exercise political jurisdiction over both dogs and public space. As a case study, we systematically collected and analyzed a local newspaper's portrayals of issues relating to dog-walking in public space. Our purpose was to understand how public and policy agendas were being represented by local media, during a period of time when local policies on dog ownership and dog-supportive public space were being reviewed and revised. The analysis involved three phases: 1) thematic content analysis; 2) issue framing analysis; and 3) policy analysis. Thematically, we found that social conflict predominated. In terms of issue framing, local policy and individual dog owner behavior were both problematized. Over time, the policy agenda evolved to emphasize the design and management of public space, especially park land. Policy-makers presented this emphasis on public space as a proactive strategy to reduce social conflict and to promote compliance with existing rules on dog ownership. When it comes to promoting dog-walking to benefit animal welfare and human health, our findings underscore the importance of paying close attention to local policies and media coverage. In fact, we found that endogenous conflict and public controversies can actually represent significant opportunities to bring about positive changes in the lives of humans and nonhuman animals.     

The Effect of Priming on Perceptions of Dog Breed Traits

ABSTRACT

This study examined whether priming cues embedded in mediastyle presentations shaped people's perceptions of specific dog breeds, and in particular, the German shepherd dog (GSD). Two hundred and four adult females were exposed to one of two types of media-style presentation (stories or pictures). Half of the participants in each condition were exposed to versions designed to portray the GSD in a positive light; the remainder to stimuli developed to present the same breed in a negative light. Participants subsequently rated six individual breeds of dog, including the target breed, on a number of traits (e.g., “friendliness,” “aggression”). Analysis revealed a significant effect of priming on people's perceptions of the GSD. Participants exposed to the negative stimuli perceived this breed as significantly less approachable, and more dangerous and aggressive, than those exposed to the positive stimuli. Priming did not influence the participants' perceptions of other breeds, even those often regarded in a negative light, although there was some evidence of breed-related category-based stereotyping. Overall, results suggest that people's perceptions of dog breeds can be influenced by verbal and visual representations. The results have implications for how dogs are portrayed in the media and other publically available sources of information.     

Liminality and racial hazing of Muslim migrants: media framing of Albanians in Shepparton,Australia, 1930–1955

This article is a historical empirical study of the Albanian Muslim migrant community of Shepparton. Through analysing newspaper reports, the authors discuss how these migrants were portrayed as liminal between their first arrival and acceptance as Australians a generation later. This is characteristic of a practice which the authors term “migrant hazing”, where a migrant group is demonized as a threat to the society during the liminal phase. Migrant hazing occurs in public discourse, particularly the media, and ceases with the replacement of the group by newer migrants, who are subjected to the same process. Furthermore, migrant hazing remains present in contemporary depictions of Australian Muslims. In this longitudinal study, media reports on Albanian Muslims revolved around three persistent themes: their supposed criminality, the wrongful use of land and the threat of dual-loyalty. These three items constituted the main weapons of the media in hazing the first, liminal generation.     

Negotiating British Muslim belonging: a qualitative longitudinal study

British Muslims are often viewed as holding values incompatible with Britishness, regarded with suspicion and sometimes subjected to gendered forms of racism. Research projects have found that identifiably Muslim women face everyday microaggressions, yet little is known about how they negotiate both this and their identities over time. This article addresses this gap by reporting the results of qualitative longitudinal research that explores the narratives of two young British Muslim women over a seven-year period. The women were first interviewed when they were single undergraduates in 2010 and followed up as married young professionals in 2017. On both occasions they were negotiating their identities and sense of belonging in a climate of heightened scrutiny of Muslims. The paper examines their reflections on: “fitting in” with Britishness, their religious identities and the complexity of belonging. Methodologically, it contributes to qualitative longitudinal narrative research.     

Continuous hydrogen peroxide production by organic buffers in phytoplankton culture media

We investigated the production of hydrogen peroxide (HOOH) in illuminated seawater media containing a variety of zwitterionic buffers. Production rates varied extensively among buffers, with 4‐(2‐hydroxyethyl)1‐piperazineethanesulfonic acid (HEPES) highest and N‐Tris(hydroxymethyl)methyl‐3‐aminopropanesulfonic acid (TAPS) among the lowest. The rate of HOOH accumulation was remarkably consistent over many days, and increased linearly with buffer concentration, natural seawater concentration, and light level. Concentrations of HEPES commonly used in culture media (1–10 mM) generated enough HOOH to kill the axenic Prochlorococcus strain VOL1 during growth in enriched seawater media at lower, environmentally realistic cell concentrations and/or under high light exposure. We also demonstrated that HEPES can be used experimentally to study the biological effects of chronic exposure to sublethal levels of HOOH such as may be experienced by light‐exposed microorganisms.     

Spatial pattern characterization of linear polarization-sensitive backscattering Mueller matrix elements of human serum albumin sphere suspension

Human serum albumin (HSA) nanometer or micron particles represent promising drug-carrier systems. The azimuthal and radial variations of a linear polarization-sensitive backscattering Mueller matrix were experimentally studied in two cases: the scattering particle was smaller or larger in size to the probing wavelength of 780 nm. The results show that the twofold and fourfold structures are characteristic of small particle size suspension, whereas the eightfold structure is characteristic of large particle size suspension. Moreover, for both particle size suspensions, the element patterns have strong radial dependence when the suspension concentration and the incident power of laser change. In addition, for both particle size suspensions, the rotational symmetry of each element is lost in the case of oblique incidence but the multifold structure is maintained. Some suggestions for applications of Mueller matrix imaging in biomedical optics are provided.