Analysis by confocal microscopy of the behavior of heat shock protein 70 within the nucleus and of a nuclear matrix polypeptide during prolonged heat shock response in HeLa cells.

By means of confocal laser scanning microscopy and indirect fluorescence experiments we have examined the behavior of heat-shock protein 70 (HSP70) within the nucleus as well as of a nuclear matrix protein (M(r) = 125 kDa) during a prolonged heat-shock response (up to 24 h at 42 degrees C) in HeLa cells. In control cells HSP70 was mainly located in the cytoplasm. The protein translocated within the nucleus upon cell exposure to hyperthermia. The fluorescent pattern revealed by monoclonal antibody to HSP70 exhibited several changes during the 24-h-long incubation. The nuclear matrix protein showed changes in its location that were evident as early as 1 h after initiation of heat shock. After 7 h of treatment, the protein regained its original distribution. However, in the late stages of the hyperthermic treatment (17-24 h) the fluorescent pattern due to 125-kDa protein changed again and its original distribution was never observed again. These results show that HSP70 changes its localization within the nucleus conceivably because it is involved in solubilizing aggregated polypeptides present in different nuclear regions. Our data also strengthen the contention that proteins of the insoluble nucleoskeleton are involved in nuclear structure changes that occur during heat-shock response.

Enhanced expression of 70-kilodalton heat shock protein limits cell division in a sepsis-induced model of acute respiratory distress syndrome.

OBJECTIVE: Fibrotic changes are initiated early in acute respiratory distress syndrome. This may involve overproliferation of alveolar type II cells. In an animal model of acute respiratory distress syndrome, we have shown that the administration of an adenoviral vector overexpressing the 70-kd heat shock protein (AdHSP) limited pathophysiological changes. We hypothesized that this improvement may be modulated, in part, by an early AdHSP-induced attenuation of alveolar type II cell proliferation. DESIGN: Laboratory investigation. SETTING: Hadassah-Hebrew University and University of Pennsylvania animal laboratories. SUBJECTS: Sprague-Dawley Rats (250 g). INTERVENTIONS: Lung injury was induced in male Sprague-Dawley rats via cecal ligation and double puncture. At the time of cecal ligation and double puncture, we injected phosphate-buffered saline, AdHSP, or AdGFP (an adenoviral vector expressing the marker green fluorescent protein) into the trachea. Rats then received subcutaneous bromodeoxyuridine. In separate experiments, A549 cells were incubated with medium, AdHSP, or AdGFP. Some cells were also stimulated with tumor necrosis factor-alpha. After 48 hrs, cytosolic and nuclear proteins from rat lungs or cell cultures were isolated. These were subjected to immunoblotting, immunoprecipitation, electrophoretic mobility shift assay, fluorescent immunohistochemistry, and Northern blot analysis. MEASUREMENTS AND MAIN RESULTS: Alveolar type I cells were lost within 48 hrs of inducing acute respiratory distress syndrome. This was accompanied by alveolar type II cell proliferation. Treatment with AdHSP preserved alveolar type I cells and limited alveolar type II cell proliferation. Heat shock protein 70 prevented overexuberant cell division, in part, by inhibiting hyperphosphorylation of the regulatory retinoblastoma protein. This prevented retinoblastoma protein ubiquitination and degradation and, thus, stabilized the interaction of retinoblastoma protein with E2F1, a key cell division transcription factor. CONCLUSIONS: : Heat shock protein 70-induced attenuation of cell proliferation may be a useful strategy for limiting lung injury when treating acute respiratory distress syndrome if consistent in later time points.

Audit report on Sheldon Community School District in Sheldon, Iowa for the year ended June 30, 2009

Audit report on Sheldon Community School District in Sheldon, Iowa for the year ended June 30, 2009

Audit report on Dunkerton Community School District in Dunkerton, Iowa for the year ended June 30, 2009

Audit report on Dunkerton Community School District in Dunkerton, Iowa for the year ended June 30, 2009

Report on a special investigation of the Cedar Rapids Community School District for the period July 1, 2002 through August 24, 2009

Report on a special investigation of the Cedar Rapids Community School District for the period July 1, 2002 through August 24, 2009

Fact Sheet: Extreme Heat, 2009

Doing too much on a hot day, spending too much time in the sun or staying too long in an overheated place can cause heat-related illnesses. Know the symptoms of heat disorders and overexposure to the sun, and be ready to give first aid treatment.

Audit report on the Disaster Grants – Public Assistance program of the Green Bay Levee and Drainage District in Lee County, Iowa for the year ended June 30, 2009

Audit report on the Disaster Grants – Public Assistance program of the Green Bay Levee and Drainage District in Lee County, Iowa for the year ended June 30, 2009

Report on a special investigation of the Food Service Department of the Springville Community School District for the period July 1, 2005 through March 31, 2009

Report on a special investigation of the Food Service Department of the Springville Community School District for the period July 1, 2005 through March 31, 2009

Audit report on the Xenia Rural Water District for the year ended December 31, 2009

Audit report on the Xenia Rural Water District for the year ended December 31, 2009

Combined report on the eight Judicial District Departments of Correctional Services for the year ended June 30, 2009

Combined report on the eight Judicial District Departments of Correctional Services for the year ended June 30, 2009

Audit report on the Disaster Grants – Public Assistance program of the Iowa River - Flint Creek Levee District in Des Moines County and Louisa County, Iowa for the year ended June 30, 2009

Audit report on the Disaster Grants – Public Assistance program of the Iowa River - Flint Creek Levee District in Des Moines County and Louisa County, Iowa for the year ended June 30, 2009

Audit report on the Disaster Grants – Public Assistance program of Louisa – Des Moines County Drainage District in Des Moines County and Louisa County, Iowa for the year ended June 30, 2009

Audit report on the Disaster Grants – Public Assistance program of Louisa – Des Moines County Drainage District in Des Moines County and Louisa County, Iowa for the year ended June 30, 2009

Report on a special investigation of the Clinton Community School District for the period July 1, 2005 through February 28, 2010

Report on a special investigation of the Clinton Community School District for the period July 1, 2005 through February 28, 2010

Report on the Iowa Judicial Branch – County Clerks of District Courts, a part of the State of Iowa, for the year ended June 30, 2009

Report on the Iowa Judicial Branch – County Clerks of District Courts, a part of the State of Iowa, for the year ended June 30, 2009

Letter to the Colo-Nesco Community School District as a result of reaudit procedures performed at the request of the Superintendent pursuant to Chapter 11.6(4)(a)(2) of the Code of Iowa for the period July 1, 2007 through June 30, 2008

Letter to the Colo-Nesco Community School District as a result of reaudit procedures performed at the request of the Superintendent pursuant to Chapter 11.6(4)(a)(2) of the Code of Iowa for the period July 1, 2007 through June 30, 2008