Hypoxia-inducible factor-1alpha is a critical mediator of hypoxia induced apoptosis in cardiac H9c2 and kidney epithelial HK-2 cells

<p>Abstract</p> <p>Background</p> <p>Hypoxia inducible factor-1 (HIF-1) is a transcription factor that functions to maintain cellular homeostasis in response to hypoxia. There is evidence that HIF-1 can also trigger apoptosis, possibly when cellular responses are inadequate to meet energy demands under hypoxic conditions.</p> <p>Methods</p> <p>Cardiac derived H9c2 and renal tubular epithelial HK-2 cells expressing either the wild type oxygen regulated subunit of HIF-1 (pcDNA3-Hif-1α) or a dominant negative version that lacked both DNA binding and transactivation domains (pcDNA3-DN-Hif-1α), were maintained in culture and exposed to hypoxia. An RNA interference approach was also employed to selectively knockdown expression of Hif-1α. Apoptosis was analyzed in both H9c2 and HK-2 cells by Hoechst and TUNEL staining, caspase 3 activity assays and activation of pro-apoptotic Bcl2 family member Bax.</p> <p>Results</p> <p>Overexpression of pcDNA3-DN-Hif-1α led to a significant reduction in hypoxia -induced apoptosis (17 ± 2%, <it>P </it>< 0.01) in H9c2 cells compared to both control-transfected and wild type Hif-1α transfected cells. Moreover, selective ablation of HIF-1α protein expression by RNA interference in H9c2 cells led to 55% reduction of caspase 3 activity and 46% reduction in the number of apoptotic cells as determined by Hoechst 33258 staining, after hypoxia. Finally, upregulation of the pro-apoptotic protein, Bax, was found in H9c2 cells overexpressing full-length pcDNA3-HA-HIF-1α exposed to hypoxia. In HK-2 cells overexpression of wild-type Hif-1α led to a two-fold increase in Hif-1α levels during hypoxia. This resulted in a 3.4-fold increase in apoptotic cells and a concomitant increase in caspase 3 activity during hypoxia when compared to vector transfected control cells. HIF-1α also induced upregulation of Bax in HK-2 cells. In addition, introduction of dominant negative Hif-1α constructs in both H9c2 and HK-2 -cells led to decreased active Bax expression.</p> <p>Conclusion</p> <p>These data demonstrate that HIF-1α is an important component of the apoptotic signaling machinery in the two cell types.</p

Eradication of Metastatic Renal Cell Carcinoma after Adenovirus-Encoded TNF-Related Apoptosis-Inducing Ligand (TRAIL)/CpG Immunotherapy

Despite evidence that antitumor immunity can be protective against renal cell carcinoma (RCC), few patients respond objectively to immunotherapy and the disease is fatal once metastases develop. We asked to what extent combinatorial immunotherapy with Adenovirus-encoded murine TNF-related apoptosis-inducing ligand (Ad5mTRAIL) plus CpG oligonucleotide, given at the primary tumor site, would prove efficacious against metastatic murine RCC. To quantitate primary renal and metastatic tumor growth in mice, we developed a luciferase-expressing Renca cell line, and monitored tumor burdens via bioluminescent imaging. Orthotopic tumor challenge gave rise to aggressive primary tumors and lung metastases that were detectable by day 7. Intra-renal administration of Ad5mTRAIL+CpG on day 7 led to an influx of effector phenotype CD4 and CD8 T cells into the kidney by day 12 and regression of established primary renal tumors. Intra-renal immunotherapy also led to systemic immune responses characterized by splenomegaly, elevated serum IgG levels, increased CD4 and CD8 T cell infiltration into the lungs, and elimination of metastatic lung tumors. Tumor regression was primarily dependent upon CD8 T cells and resulted in prolonged survival of treated mice. Thus, local administration of Ad5mTRAIL+CpG at the primary tumor site can initiate CD8-dependent systemic immunity that is sufficient to cause regression of metastatic lung tumors. A similar approach may prove beneficial for patients with metastatic RCC

Estimating the post-closure management time for landfills containing treated MSW residues.

The Landfill Directive will require the pre-treatment of MSW prior to landfilling. The removal of progressively increasing proportions of the biodegradable fractions from landfilled waste, and the UK Government’s commitment to increase recycling of key waste fractions, will lead to an inevitable change, from the disposal of raw MSW, to the disposal of MSW treatment residues, to landfill. This will undoubtedly change the type and rate of emissions from landfills. The question that this research project has sought to answer is “how long will active management be required for different MSW treatment residues?” The term equilibrium status has been used to define the end point beyond which management of wastes is no longer necessary. Calculating the equilibrium status of waste involves an assessment of the landfill gas emissions, leachate quality and hydraulic status of the landfill. These key parameters change with time as the landfill evolves. Equally, the engineering performance of a site is also changing with the gradual degradation of the liner and capping systems. The question posed above is therefore not easily answered

Estimating Pollutant Removal Requirements for Landfills in the UK: II. Model Development

A modelling methodology using a leachate source term has been produced for estimating the timescales for achieving environmental equilibrium status for landfilled waste. Results are reported as the period of active management required for modelled scenarios of non-flushed and flushed sites for a range of pre-filling treatments. The base scenario against which results were evaluated was raw municipal solid waste (MSW) for which only cadmium failed to reach equilibrium. Flushed raw MSW met our criteria for stabilisation with active leachate management for 40 years, subject to each of the leachate species being present at or below their average UK concentrations. Stable non-reactive wastes, meeting EU waste acceptance criteria, fared badly in the non-flushed scenario, with only two species stabilising after a management period within 1000 years and the majority requiring >2000 years of active leachate management. The flushing scenarios showed only a marginal improvement, with arsenic still persisting beyond 2000 years management even with an additional 500 mm y−1 of infiltration. The stabilisation time for mechanically sorted organic residues (without flushing) was high, and even with flushing, arsenic and chromium appeared to remain a problem. Two mechanical biological treatment (MBT) scenarios were examined, with medium and high intensity composting. Both were subjected to the non-flushing and flushing scenarios. The non-flushing case of both options fell short of the basic requirements of achieving equilibrium within decades. The intense composting option with minimal flushing appeared to create a scenario where equilibrium could be achieved. For incinerator bottom ash (raw and subjected to various treatments), antimony, copper, chloride and sulphate were the main controls on achieving equilibrium, irrespective of treatment type. Flushing at higher flushing rates (500 mm y−1) failed to demonstrate a significant reduction in the management period requ

Estimating Pollutant Removal Requirements for Landfills in the UK: I. Benchmark Study and Characteristics of Waste Treatment Technologies

Introduction of the EU Landfill Directive is having a significant impact on waste management in the UK and in other member states that have relied on landfilling. This paper considers the length of the aftercare period required by the municipal solid waste streams that the UK will most probably generate following implementation of the Landfill Directive. Data were derived from literature to identify properties of residues from the most likely treatment processes and the probable management times these residues will require within the landfill environment were then modelled. Results suggest that for chloride the relevant water quality standard (250 mg l−1) will be achieved with a management period of 40 years and for lead (0.1 mg l−1), 240 years. This has considerable implications for the sustainability of landfill and suggests that current timescales for aftercare of landfills may be inadeq

Vegetation map of Nigeria /

First edition - F.N. "F.N. 1."; "1800/406/4-59."; Also available online http://nla.gov.au/nla.map-vn6807116

The carnivores of West Africa /

Includes index.Bibliography: p. 524-537