The mode of lymphoblastoid cell death in response to gas phase cigarette smoke is dose-dependent

<p>Abstract</p> <p>Background</p> <p>Cigarette smoke (CS) is the main cause in the development of chronic obstructive pulmonary disease (COPD), the pathogenesis of which is related to an extended inflammatory response. In this study, we investigated the effect of low and high doses of gas phase cigarette smoke (GPS) on cultured lymphocyte progenitor cells, using techniques to assess cell viability and to elucidate whether cells die of apoptosis or necrosis upon exposure to different doses of GPS.</p> <p>Methods</p> <p>In our approach we utilised a newly-established system of exposure of cells to GPS that is highly controlled, accurately reproducible and simulates CS dosage and kinetics that take place in the smokers' lung. This system was used to study the mode of cell death upon exposure to GPS in conjunction with a range of techniques widely used for cell death studies such as Annexin V staining, activation of caspase -3, cytoplasmic release of cytochrome C, loss of mitochondrial membrane potential and DNA fragmentation.</p> <p>Results</p> <p>Low doses of GPS induced specific apoptotic indexes in CCRF-CEM cells. Specifically, cytochrome C release and cleaved caspase-3 were detected by immunofluorescence, upon treatment with 1-3 puffs GPS. At 4 h post-exposure, caspase-3 activation was observed in western blot analysis, showing a decreasing pattern as GPS doses increased. Concomitant with this behaviour, a dose-dependent change in Δψ_mdepolarization was monitored by flow cytometry 2 h post-exposure, while at 4 h Δψ_mcollapse was observed at the higher doses, indicative of a shift to a necrotic demise. A reduction in DNA fragmentation events produced by 5 puffs GPS as compared to those provoked by 3 puffs GPS, also pointed towards a necrotic response at the higher dose of GPS.</p> <p>Conclusion</p> <p>Collectively, our results support that at low doses gas phase cigarette smoke induces apoptosis in cultured T-lymphocytes, whereas at high doses GPS leads to necrotic death, by-passing the characteristic stage of caspase-3 activation and, thus, the apoptotic route.</p

Mind the gaps! A research agenda for urban interstices

Processes of urbanisation can hardly be considered without reference to the spaces that lie between developments. However, the literature on such interstitial spaces is fragmentary. In this paper we draw together insights from the extant literature into a research agenda on urban interstices. We propose a research agenda centred on four themes: the multiple geographic scales at which the interstitial spaces of urban sprawl might be analysed; the pending nature of such spaces; their planned or unplanned character and their relational properties. We develop these themes, briefly illustrating them with reference to the case of metropolitan area of Santiago de Chile. In conclusion, we emphasise some of the implications of interstitial spaces for theories of urban politics and their value in forcing inter-disciplinarity in urban studies

Organizing solidarity initiatives : a socio-spatial conceptualization of resistance

This paper offers a spatial conceptualization of resistance by focusing on the practices through which solidarity initiatives constitute new resistance socio-spatialities. We discuss two solidarity initiatives in Greece, WCNA and Vio.Me.SI, and explore how they institute distinctive local and translocal organizational practices that make the production of new forms of resistance possible. In particular, we adopt a productive and transformative view of resistance. First, we identify three local practices of organizing solidarity initiatives, namely, the organization of general assembly meetings, the constitution of resistance laboratories and the (re)articulation of socio-spatial relations in local sites. Then, we turn to flows, movements and translocal social formations, and examine the role of solidarity mobilizations, the material and symbolic co-production of resources and members’ mobility in the production of resistance. We conclude that new resistance socio-spatialities become constitutive of a broader reconfiguration of political agencies, a creative process that challenges existing relations and invites alternative ways of working and organizing

X-linked dyskeratosis congenita is predominantly caused by missense mutations in the DKC1 gene.

Dyskeratosis congenita is a rare inherited bone marrow-failure syndrome characterized by abnormal skin pigmentation, nail dystrophy, and mucosal leukoplakia. More than 80% of patients develop bone-marrow failure, and this is the major cause of premature death. The X-linked form of the disease (MIM 305000) has been shown to be caused by mutations in the DKC1 gene. The gene encodes a 514-amino-acid protein, dyskerin, that is homologous to Saccharomyces cerevisiae Cbf5p and rat Nap57 proteins. By analogy to the homologues in other species, dyskerin is predicted to be a nucleolar protein with a role in both the biogenesis of ribosomes and, in particular, the pseudouridylation of rRNA precursors. We have determined the genomic structure of the DKC1 gene; it consists of 15 exons spanning a region of 15 kb. This has enabled us to screen for mutations in the genomic DNA, by using SSCP analysis. Mutations were detected in 21 of 37 additional families with dyskeratosis congenita that were analyzed. These mutations consisted of 11 different single-nucleotide substitutions, which resulted in 10 missense mutations and 1 putative splicing mutation within an intron. The missense change A353V was observed in 10 different families and was shown to be a recurring de novo event. Two polymorphisms were also detected, one of which resulted in the insertion of an additional lysine in the carboxy-terminal polylysine domain. It is apparent that X-linked dyskeratosis congenita is predominantly caused by missense mutations; the precise effect on the function of dyskerin remains to be determined

Investigation of heavy metal content of Turkish tobacco leaves, cigarette butt, ash, and smoke

WOS: 000325116500053PubMed ID: 23712460A procedure for the determination of cadmium, copper, manganese, and zinc in Turkish tobaccos, which were of different origins, years, and grades, and in the butt, ash, and smoke, which were obtained by smoking the cigarettes that were prepared manually from the said tobaccos in a smoking apparatus, was devised as proposed. The collected samples were digested by wet ashing technique by using HNO3-HClO4 and were analyzed by flame atomic absorption spectrometry with satisfactory recoveries (94 % to 98 %). The regression coefficients were above 0.99, and the detection limits were in the range of 0.03-0.12 mg/L-1. The performance and accuracy of the method was tested by analyzing "Certified Reference Material GBW 08501-Peach Leaves." The determined values were in agreement with the standard values for the heavy metals analyzed. Thus, it was concluded that the developed method could offer a wide range of application for establishing a relationship between the makeup and composition of tobacco plant, products, ash, smoke, and smoking