Soluble histone H2AX is induced by DNA replication stress and sensitizes cells to undergo apoptosis

<p>Abstract</p> <p>Background</p> <p>Chromatin-associated histone H2AX is a key regulator of the cellular responses to DNA damage. However, non-nucleosomal functions of histone H2AX are poorly characterized. We have recently shown that soluble H2AX can trigger apoptosis but the mechanisms leading to non-chromatin-associated H2AX are unclear. Here, we tested whether stalling of DNA replication, a common event in cancer cells and the underlying mechanism of various chemotherapeutic agents, can trigger increased soluble H2AX.</p> <p>Results</p> <p>Transient overexpression of H2AX was found to lead to a detectable fraction of soluble H2AX and was associated with increased apoptosis. This effect was enhanced by the induction of DNA replication stress using the DNA polymerase α inhibitor aphidicolin. Cells manipulated to stably express H2AX did not contain soluble H2AX, however, short-term treatment with aphidicolin (1 h) resulted in detectable amounts of H2AX in the soluble nuclear fraction and enhanced apoptosis. Similarly, soluble endogenous H2AX was detected under these conditions. We found that excessive soluble H2AX causes chromatin aggregation and inhibition of ongoing gene transcription as evidenced by the redistribution and/or loss of active RNA polymerase II as well as the transcriptional co-activators CBP and p300.</p> <p>Conclusion</p> <p>Taken together, these results show that DNA replication stress rapidly leads to increased soluble H2AX and that non-chromatin-associated H2AX can sensitize cells to undergo apoptosis. Our findings encourage further studies to explore H2AX and the cellular pathways that control its expression as anti-cancer drug targets.</p

Computational Simulations of a Mach 0.745 Transonic Truss-Braced Wing Design

A joint effort between the NASA Ames and Langley Research Centers was undertaken to analyze the Mach 0.745 variant of the Boeing Transonic Truss-Braced Wing (TTBW) Design. Two different flow solvers, LAVA and USM3D, were used to predict the TTBW flight performance. Sensitivity studies related to mesh resolution and numerical schemes were conducted to define best practices for this type of geometry and flow regime. Validation efforts compared the numerical simulation results of various modeling methods against experimental data taken from the NASA Ames 11-foot Unitary Wind Tunnel experimental data. The fidelity of the computational representation of the wind tunnel experiment, such as utilizing a porous wall boundary condition to model the ventilated test section, was varied to examine how different tunnel effects influence CFD predictions. LAVA and USM3D results both show an approximate 0.5 angle of attack shift from experimental lift curve data. This drove an investigation that revealed that the trailing edge of the experimental model was rounded in comparison to the CAD model, due to manufacturing tolerances, which had not been accounted for in the initial simulations of the experiment. Simulating the TTBW with an approximation of this rounded trailing-edge reduces error by approximately 60%. An accurate representation of the tested TTBW geometry, ideally including any wing twists and deflections experienced during the test under various loading conditions, will be necessary for proper validation of the CFD

Elevated pretreatment serum levels of soluble vascular cell adhesion molecule 1 and lactate dehydrogenase as predictors of survival in cutaneous metastatic malignant melanoma.

Very rapid progression of disease with a median survival of 6-9 months is a common feature of metastatic cutaneous malignant melanoma. Nevertheless, substantial variability of survival suggests that metastatic cutaneous malignant melanoma can be divided into several biological subgroups. Pretreatment serum levels of soluble adhesion molecules and various clinical parameters in cutaneous metastatic malignant melanoma were evaluated to determine their prognostic value. In this study pretreatment serum levels of soluble vascular cell adhesion molecule 1 (sVCAM-1), soluble intercellular cell adhesion molecule 1 (sICAM-1), soluble endothelial leukocyte adhesion molecule 1 (sE-selectin) and multiple clinical factors were assessed in relation to overall survival of 97 consecutive patients with metastatic cutaneous malignant melanoma seen at our institution between May 1990 and April 1996. For statistical analysis, both univariate and multivariate Cox proportional-hazards models were used. Elevated pretreatment serum levels of sVCAM-1 (P < 0.005) and of lactate dehydrogenase (P < 0.002) were rendered statistically independent and were significantly associated with unfavourable outcome. Patients were assigned to one of three risk categories (low, intermediate and high) according to a cumulative risk score defined as the function of the sum of these two variables. There were significant differences in overall survival (P < 0.0001) between low- (n = 53, 5-year survival probability of 23.3%), intermediate- (n = 29, 5-year survival probability of 9.9%) and high-risk (n = 15) patients. Elevated pretreatment serum levels of sVCAM-1 and of lactate dehydrogenase correlate with poor outcome in metastatic cutaneous malignant melanoma. These data support risk stratification for future therapeutic trials and identify factors that need to be validated in prospective studies and may potentially influence decision-making in palliative management of patients with disseminated cutaneous malignant melanoma

Elevated serum levels of S100 and survival in metastatic malignant melanoma.

Current reports suggest serum S100 as a prognostic marker for disease progression in advanced malignant melanoma. In this study, we assessed serum levels of S100 and multiple clinical factors in relation to overall survival in 99 patients with metastatic malignant melanoma seen at our institution between May 1990 and April 1996. For statistical analysis, we used both univariate and multivariate Cox proportional-hazards models. Elevated serum levels of S100 correlated with poor outcome in metastatic malignant melanoma (P < 0.0001), univariate analysis). Upon multivariate analysis, however, S100 added no information to known clinical prognostic parameters

Establishing Best Practices for X-57 Maxwell CFD Database Generation

The X-57 Maxwell is NASAs latest electric airplane concept that has been simulated for aerodynamic performance using the structured overset and unstructured grid solvers within the Launch Ascent and Vehicle Aerodynamics (LAVA) solver framework as well as the unstructured polyhedral grid solver in Star-CCM+ for code-to-code comparison. In order to validate the predictions, comparisons were made between the CFD solutions and experimental data collected in the 12-foot Low-Speed Wind Tunnel at NASA Langley Research Center. The simulations are in preparation for the development of a comprehensive aerodynamic database which will assess aircraft performance at a variety of conditions. The findings from these simulations will establish the best practices for mesh resolution, numerical discretization, and turbulence modeling to be used for this database. Preliminary database results have shown that best-practices learned from the initial validation simulations will potentially reduce error in X-57 aerodynamic loads and moments relative to experiment by up to 14%

Nurr1 Modulation Mediates Neuroprotective Effects of Statins

The ligand-sensing transcription factor Nurr1 emerges as a promising therapeutic target for neurodegenerative pathologies but Nurr1 ligands for functional studies and therapeutic validation are lacking. Here pronounced Nurr1 modulation by statins for which clinically relevant neuroprotective effects are demonstrated, is reported. Several statins directly affect Nurr1 activity in cellular and cell-free settings with low micromolar to sub-micromolar potencies. Simvastatin as example exhibits anti-inflammatory effects in astrocytes, which are abrogated by Nurr1 knockdown. Differential gene expression analysis in native and Nurr1-silenced cells reveals strong proinflammatory effects of Nurr1 knockdown while simvastatin treatment induces several neuroprotective mechanisms via Nurr1 involving changes in inflammatory, metabolic and cell cycle gene expression. Further in vitro evaluation confirms reduced inflammatory response, improved glucose metabolism, and cell cycle inhibition of simvastatin-treated neuronal cells. These findings suggest Nurr1 involvement in the well-documented but mechanistically elusive neuroprotection by statins

Identification of genes encoding squalestatin S1 biosynthesis and in vitro production of new squalestatin analogues

A gene cluster responsible for the biosynthesis of squalestatin S1 (SQS1, 1) was identified by full genome sequencing of two SQS1-producing ascomycetes: Phoma sp. C2932 and unidentified fungus MF5453. A transformation protocol was established and a subsequent knockout of one PKS gene from the cluster led to loss of SQS1 production and enhanced concentration of an SQS1 precursor. An acyltransferase gene from the cluster was expressed in E. coli and the expressed protein MfM4 shown to be responsible for loading acyl groups from CoA onto the squalestatin core as the final step of biosynthesis. MfM4 appears to have a broad substrate selectivity for its acyl CoA substrate, allowing the in vitro synthesis of novel squalestatins

XRCC2 R188H (rs3218536), XRCC3 T241M (rs861539) and R243H (rs77381814) single nucleotide polymorphisms in cervical cancer risk

Human Papillomavirus (HPV) is the main cause of cervical cancer and its precursor lesions. Transformation may be induced by several mechanisms, including oncogene activation and genome instability. Individual differences in DNA damage recognition and repair have been hypothesized to influence cervical cancer risk. The aim of this study was to evaluate whether the double strand break gene polymorphisms XRCC2 R188H G>A (rs3218536), XRCC3 T241M C>T (rs861539) and R243H G>A (rs77381814) are associated to cervical cancer in Argentine women. A case control study consisting of 322 samples (205 cases and 117 controls) was carried out. HPV DNA detection was performed by PCR and genotyping of positive samples by EIA (enzyme immunoassay). XRCC2 and 3 polymorphisms were determined by pyrosequencing. The HPV-adjusted odds ratio (OR) of XRCC2 188 GG/AG genotypes was OR = 2.4 (CI = 1.1-4.9, p = 0.02) for cervical cancer. In contrast, there was no increased risk for cervical cancer with XRCC3 241 TT/CC genotypes (OR = 0.48; CI = 0.2-1; p = 0.1) or XRCC3 241 CT/CC (OR = 0.87; CI = 0.52-1.4; p = 0.6). Regarding XRCC3 R243H, the G allele was almost fixed in the population studied. In conclusion, although the sample size was modest, the present data indicate a statistical association between cervical cancer and XRCC2 R188H polymorphism. Future studies are needed to confirm these findings.Fil: Perez, Luis Orlando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; ArgentinaFil: Crivaro, Andrea Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; ArgentinaFil: Barbisan, Gisela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; ArgentinaFil: Poleri, Lucía Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; ArgentinaFil: Golijow, Carlos Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentin

Computational Simulations of a Mach 0.745 Transonic Truss-Braced Wing Design

A joint effort between the NASA Ames and Langley Research Centers was undertaken to analyze the Mach 0.745 variant of the Boeing Transonic Truss-Braced Wing (TTBW) Design. Two different flow solvers, LAVA and USM3D, were used to predict the TTBW flight performance. Sensitivity studies related to mesh resolution and numerical schemes were conducted to define best practices for this type of geometry and flow regime. Validation efforts compared the numerical simulation results of various modeling methods against experimental data taken from the NASA Ames 11-foot Unitary Wind Tunnel experimental data. The fidelity of the computational representation of the wind tunnel experiment, such as utilizing a porous wall boundary condition to model the ventilated test section, was varied to examine how different tunnel effects influence CFD predictions. LAVA and USM3D results both show an approximate 0.5o angle of attack shift from experimental lift curve data. This drove an investigation that revealed that the trailing edge of the experimental model was rounded in comparison to the CAD model, due to manufacturing tolerances, which had not been accounted for in the initial simulations of the experiment. Simulating the TTBW with an approximation of this rounded trailing-edge reduces error by approximately 60%. An accurate representation of the tested TTBW geometry, ideally including any wing twists and deflections experienced during the test under various loading conditions, will be necessary for more thorough validation of the CFD

Computational Simulations of a Mach 0.745 Transonic Truss-Braced Wing Design

No abstract availabl