Using the XMM Optical Monitor to Study Cluster Galaxy Evolution

We explore the application of XMM-Newton Optical Monitor (XMM-OM) ultraviolet (UV) data to study galaxy evolution. Our sample is constructed as the intersection of all Abell clusters with z < 0.05 and having archival XMM-OM data in either the UVM2 or UVW1 filters, plus optical and UV photometry from the Sloan Digital Sky Survey and GALEX, respectively. The eleven resulting clusters include 726 galaxies with measured redshifts, 520 of which have redshifts placing them within their parent Abell clusters. We develop procedures for manipulating the XMM-OM images and measuring galaxy photometry from them, and confirm our results via comparison with published catalogs. Color magnitude diagrams (CMDs) constructed using the XMM-OM data along with SDSS optical data show promise for evolutionary studies, with good separation between red and blue sequences and real variation in the width of the red sequence that is likely indicative of differences in star formation history. This is particularly true for UVW1 data, as the relative abundance of data collected using this filter and its depth make it an attractive choice. Available tools that use stellar synthesis libraries to fit the UV and optical photometric data may also be used, thereby better describing star formation history within the past Gyr and providing estimates of total stellar mass that include contributions from young stars. Finally, color-color diagrams that include XMM-OM UV data appear useful to the photometric identification of both extragalactic and stellar sources.Comment: 44 pages with 14 figures, to appear in PAS

Nitrosative stress induces DNA strand breaks but not caspase mediated apoptosis in a lung cancer cell line

BACKGROUND: Key steps crucial to the process of tumor progression are genomic instability and escape from apoptosis. Nitric oxide and its interrelated reactive intermediates (collectively denoted as NO(X)) have been implicated in DNA damage and mutational events leading to cancer development, while also being implicated in the inhibition of apoptosis through S-nitrosation of key apoptotic enzymes. The purpose of this study was to explore the interrelationship between NO(X)-mediated DNA strand breaks (DSBs) and apoptosis in cultured tumor cell lines. METHODS: Two well-characterized cell lines were exposed to increasing concentrations of exogenous NO(X )via donor compounds. Production of NO(X )was quantified by the Greiss reaction and spectrophotometery, and confirmed by nitrotyrosine immunostaining. DSBs were measured by the alkaline single-cell gel electrophoresis assay (the COMET assay), and correlated with cell viability by the MTT assay. Apoptosis was analyzed both by TUNEL staining and Annexin V/propidium iodine FACS. Finally, caspase enzymatic activity was measured using an in-vitro fluorogenic caspase assay. RESULTS: Increases in DNA strand breaks in our tumor cells, but not in control fibroblasts, correlated with the concentration as well as rate of release of exogenously administered NO(X). This increase in DSBs did not correlate with an increase in cell death or apoptosis in our tumor cell line. Finally, this lack of apoptosis was found to correlate with inhibition of caspase activity upon exposure to thiol- but not NONOate-based NO(X )donor compounds. CONCLUSIONS: Genotoxicity appears to be highly interrelated with both the concentration and kinetic delivery of NO(X). Moreover, alterations in cell apoptosis can be seen as a consequence of the explicit mechanisms of NO(X )delivery. These findings lend credence to the hypothesis that NO(X )may play an important role in tumor progression, and underscores potential pitfalls which should be considered when developing NO(X)-based chemotherapeutic agents

Bacterial Hypoxic Responses Revealed as Critical Determinants of the Host-Pathogen Outcome by TnSeq Analysis of Staphylococcus aureus Invasive Infection

Staphylococcus aureus is capable of infecting nearly every organ in the human body. In order to infiltrate and thrive in such diverse host tissues, staphylococci must possess remarkable flexibility in both metabolic and virulence programs. To investigate the genetic requirements for bacterial survival during invasive infection, we performed a transposon sequencing (TnSeq) analysis of S. aureus during experimental osteomyelitis. TnSeq identified 65 genes essential for staphylococcal survival in infected bone and an additional 148 mutants with compromised fitness in vivo. Among the loci essential for in vivo survival was SrrAB, a staphylococcal two-component system previously reported to coordinate hypoxic and nitrosative stress responses in vitro. Healthy bone is intrinsically hypoxic, and intravital oxygen monitoring revealed further decreases in skeletal oxygen concentrations upon S. aureus infection. The fitness of an srrAB mutant during osteomyelitis was significantly increased by depletion of neutrophils, suggesting that neutrophils impose hypoxic and/or nitrosative stresses on invading bacteria. To more globally evaluate staphylococcal responses to changing oxygenation, we examined quorum sensing and virulence factor production in staphylococci grown under aerobic or hypoxic conditions. Hypoxic growth resulted in a profound increase in quorum sensing-dependent toxin production, and a concomitant increase in cytotoxicity toward mammalian cells. Moreover, aerobic growth limited quorum sensing and cytotoxicity in an SrrAB-dependent manner, suggesting a mechanism by which S. aureus modulates quorum sensing and toxin production in response to environmental oxygenation. Collectively, our results demonstrate that bacterial hypoxic responses are key determinants of the staphylococcal-host interaction

Dynamic Modeling of Cell Migration and Spreading Behaviors on Fibronectin Coated Planar Substrates and Micropatterned Geometries

An integrative cell migration model incorporating focal adhesion (FA) dynamics, cytoskeleton and nucleus remodeling, actin motor activity, and lamellipodia protrusion is developed for predicting cell spreading and migration behaviors. This work is motivated by two experimental works: (1) cell migration on 2-D substrates under various fibronectin concentrations and (2) cell spreading on 2-D micropatterned geometries. These works suggest (1) cell migration speed takes a maximum at a particular ligand density (~1140 molecules/µm2) and (2) that strong traction forces at the corners of the patterns may exist due to combined effects exerted by actin stress fibers (SFs). The integrative model of this paper successfully reproduced these experimental results and indicates the mechanism of cell migration and spreading. In this paper, the mechanical structure of the cell is modeled as having two elastic membranes: an outer cell membrane and an inner nuclear membrane. The two elastic membranes are connected by SFs, which are extended from focal adhesions on the cortical surface to the nuclear membrane. In addition, the model also includes ventral SFs bridging two focal adhesions on the cell surface. The cell deforms and gains traction as transmembrane integrins distributed over the outer cell membrane bond to ligands on the ECM surface, activate SFs, and form focal adhesions. The relationship between the cell migration speed and fibronectin concentration agrees with existing experimental data for Chinese hamster ovary (CHO) cell migrations on fibronectin coated surfaces. In addition, the integrated model is validated by showing persistent high stress concentrations at sharp geometrically patterned edges. This model will be used as a predictive model to assist in design and data processing of upcoming microfluidic cell migration assays

Elective Cancer Surgery in COVID-19-Free Surgical Pathways During the SARS-CoV-2 Pandemic: An International, Multicenter, Comparative Cohort Study.

PURPOSE: As cancer surgery restarts after the first COVID-19 wave, health care providers urgently require data to determine where elective surgery is best performed. This study aimed to determine whether COVID-19-free surgical pathways were associated with lower postoperative pulmonary complication rates compared with hospitals with no defined pathway. PATIENTS AND METHODS: This international, multicenter cohort study included patients who underwent elective surgery for 10 solid cancer types without preoperative suspicion of SARS-CoV-2. Participating hospitals included patients from local emergence of SARS-CoV-2 until April 19, 2020. At the time of surgery, hospitals were defined as having a COVID-19-free surgical pathway (complete segregation of the operating theater, critical care, and inpatient ward areas) or no defined pathway (incomplete or no segregation, areas shared with patients with COVID-19). The primary outcome was 30-day postoperative pulmonary complications (pneumonia, acute respiratory distress syndrome, unexpected ventilation). RESULTS: Of 9,171 patients from 447 hospitals in 55 countries, 2,481 were operated on in COVID-19-free surgical pathways. Patients who underwent surgery within COVID-19-free surgical pathways were younger with fewer comorbidities than those in hospitals with no defined pathway but with similar proportions of major surgery. After adjustment, pulmonary complication rates were lower with COVID-19-free surgical pathways (2.2% v 4.9%; adjusted odds ratio [aOR], 0.62; 95% CI, 0.44 to 0.86). This was consistent in sensitivity analyses for low-risk patients (American Society of Anesthesiologists grade 1/2), propensity score-matched models, and patients with negative SARS-CoV-2 preoperative tests. The postoperative SARS-CoV-2 infection rate was also lower in COVID-19-free surgical pathways (2.1% v 3.6%; aOR, 0.53; 95% CI, 0.36 to 0.76). CONCLUSION: Within available resources, dedicated COVID-19-free surgical pathways should be established to provide safe elective cancer surgery during current and before future SARS-CoV-2 outbreaks

Elective cancer surgery in COVID-19-free surgical pathways during the SARS-CoV-2 pandemic: An international, multicenter, comparative cohort study

PURPOSE As cancer surgery restarts after the first COVID-19 wave, health care providers urgently require data to determine where elective surgery is best performed. This study aimed to determine whether COVID-19–free surgical pathways were associated with lower postoperative pulmonary complication rates compared with hospitals with no defined pathway. PATIENTS AND METHODS This international, multicenter cohort study included patients who underwent elective surgery for 10 solid cancer types without preoperative suspicion of SARS-CoV-2. Participating hospitals included patients from local emergence of SARS-CoV-2 until April 19, 2020. At the time of surgery, hospitals were defined as having a COVID-19–free surgical pathway (complete segregation of the operating theater, critical care, and inpatient ward areas) or no defined pathway (incomplete or no segregation, areas shared with patients with COVID-19). The primary outcome was 30-day postoperative pulmonary complications (pneumonia, acute respiratory distress syndrome, unexpected ventilation). RESULTS Of 9,171 patients from 447 hospitals in 55 countries, 2,481 were operated on in COVID-19–free surgical pathways. Patients who underwent surgery within COVID-19–free surgical pathways were younger with fewer comorbidities than those in hospitals with no defined pathway but with similar proportions of major surgery. After adjustment, pulmonary complication rates were lower with COVID-19–free surgical pathways (2.2% v 4.9%; adjusted odds ratio [aOR], 0.62; 95% CI, 0.44 to 0.86). This was consistent in sensitivity analyses for low-risk patients (American Society of Anesthesiologists grade 1/2), propensity score–matched models, and patients with negative SARS-CoV-2 preoperative tests. The postoperative SARS-CoV-2 infection rate was also lower in COVID-19–free surgical pathways (2.1% v 3.6%; aOR, 0.53; 95% CI, 0.36 to 0.76). CONCLUSION Within available resources, dedicated COVID-19–free surgical pathways should be established to provide safe elective cancer surgery during current and before future SARS-CoV-2 outbreaks

Deficient prepulse inhibition in schizophrenia detected by the multi-site COGS

BACKGROUND: Startle inhibition by weak prepulses (PPI) is studied to understand the biology of information processing in schizophrenia patients and healthy comparison subjects (HCS). The Consortium on the Genetics of Schizophrenia (COGS) identified associations between PPI and single nucleotide polymorphisms in schizophrenia probands and unaffected relatives, and linkage analyses extended evidence for the genetics of PPI deficits in schizophrenia in the COGS-1 family study. These findings are being extended in a 5-site “COGS-2” study of 1800 patients and 1200 unrelated HCS to facilitate genetic analyses. We describe a planned interim analysis of COGS-2 PPI data. METHODS: Eyeblink startle was measured in carefully screened HCS and schizophrenia patients (n=1402). Planned analyses of PPI (60 ms intervals) assessed effects of diagnosis, sex and test site, PPI-modifying effects of medications and smoking, and relationships between PPI and neurocognitive measures. RESULTS: 884 subjects met strict inclusion criteria. ANOVA of PPI revealed significant effects of diagnosis (p=0.0005) and sex (p<0.002), and a significant diagnosis × test site interaction. HCS > schizophrenia PPI differences were greatest among patients not taking 2(nd) generation antipsychotics, and were independent of smoking status. Modest but significant relationships were detected between PPI and performance in specific neurocognitive measures. DISCUSSION: The COGS-2 multi-site study detects schizophrenia-related PPI deficits reported in single-site studies, including patterns related to diagnosis, prepulse interval, sex, medication and other neurocognitive measures. Site differences were detected and explored. The target COGS-2 schizophrenia “endophenotype” of reduced PPI should prove valuable for identifying and confirming schizophrenia risk genes in future analyses

Characterization of the Curli Nucleator CsgB.

Amyloid biogenesis is the underlying cause of many neurological disorders such as Alzheimer’s disease. During the progression of Alzheimer’s disease it is thought that normally soluble Aβ peptides self assemble into β-sheet rich, detergent resistant amyloid fibers via a template mediated-nucleation dependent polymerization reaction. Although the polymerization of Aβ is well characterized in vitro, the molecular events that govern nucleation in vivo are poorly described. Curli fibers are extracellular functional amyloid fibers utilized by enteric bacteria for the formation of biofilms. The fibers are composed of two proteins, CsgA, the major subunit, and CsgB, the minor subunit. CsgB is required for nucleating CsgA polymerization in vivo. CsgA and CsgB are 50% similar at the amino level and in silico structural predictions suggest both proteins are β-sheet rich. We hypothesized that CsgB drives the polymerization of CsgA by acting as a β-sheet rich folding template at the cell surface, and predicted that CsgB could self-polymerize into amyloid fibers in vitro. We found that CsgB and mutant variants polymerized into amyloid fibers in vitro, and that these fibers seeded CsgA polymerization. Perturbing the C-terminal portion of CsgB caused CsgB to be mislocalized in vivo and greatly reduced CsgB function. Our results are consistent with the hypothesis that CsgB templates CsgA polymerization by adopting a β-sheet rich fold, and that disruption of the CsgB-cell surface interaction leads to impaired CsgB function. Using the methods established in the course of this work, I have also begun to identify protein- and small molecule inhibitors of curli nucleation and polymerization. For example, CsgE, an accessory protein required for curli biogenesis, and rationally-designed small molecule inhibitors of Aβ polymerization inhibit CsgA polymerization in vitro. Together these results suggest curli biogenesis may be a powerful model for elucidating the initial molecular events involved in amyloid biogenesis.Ph.D.Microbiology & ImmunologyUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/63820/1/nhammer_1.pd

Using the XMM-Newton Optical Monitor to Study Cluster Galaxy Evolution

We explore the application of XMM Newton Optical Monitor (XMM-OM) ultraviolet (UV) data to study galaxy evolution. Our sample is constructed as the intersection of all Abell clusters with z < 0.05 and having archival XMM-OM data in either the UVM2 or UVW1 filters, plus optical and UV photometry from the Sloan Digital Sky Survey and GALEX, respectively. The 11 resulting clusters include 726 galaxies with measured redshifts, 520 of which have redshifts placing them within their parent Abell clusters. We develop procedures for manipulating the XMM-OM images and measuring galaxy photometry from them, and we confirm our results via comparison with published catalogs. Color-magnitude diagrams (CMDs) constructed using the XMM-OM data along with SDSS optical data show promise for evolutionary studies, with good separation between red and blue sequences and real variation in the width of the red sequence that is likely indicative of differences in star formation history. This is particularly true for UVW1 data, as the relative abundance of data collected using this filter and its depth make it an attractive choice. Available tools that use stellar synthesis libraries to fit the UV and optical photometric data may also be used, thereby better describing star formation history within the past billion years and providing estimates of total stellar mass that include contributions from young stars. Finally, color-color diagrams that include XMM-OM UV data appear useful to the photometric identification of both extragalactic and stellar sources