Proteomic Analysis of a Noninvasive Human Model of Acute Inflammation and Its Resolution: The Twenty-one Day Gingivitis Model

The 21-day experimental gingivitis model, an established noninvasive model of inflammation in response to increasing bacterial accumulation in humans, is designed to enable the study of both the induction and resolution of inflammation. Here, we have analyzed gingival crevicular fluid, an oral fluid comprising a serum transudate and tissue exudates, by LC−MS/MS using Fourier transform ion cyclotron resonance mass spectrometry and iTRAQ isobaric mass tags, to establish meta-proteomic profiles of inflammation-induced changes in proteins in healthy young volunteers. Across the course of experimentally induced gingivitis, we identified 16 bacterial and 186 human proteins. Although abundances of the bacterial proteins identified did not vary temporally, Fusobacterium outer membrane proteins were detected. Fusobacterium species have previously been associated with periodontal health or disease. The human proteins identified spanned a wide range of compartments (both extracellular and intracellular) and functions, including serum proteins, proteins displaying antibacterial properties, and proteins with functions associated with cellular transcription, DNA binding, the cytoskeleton, cell adhesion, and cilia. PolySNAP3 clustering software was used in a multilayered analytical approach. Clusters of proteins that associated with changes to the clinical parameters included neuronal and synapse associated proteins

A historical and proteomic analysis of botulinum neurotoxin type/G

<p>Abstract</p> <p>Background</p> <p><it>Clostridium botulinum </it>is the taxonomic designation for at least six diverse species that produce botulinum neurotoxins (BoNTs). There are seven known serotypes of BoNTs (/A through/G), all of which are potent toxins classified as category A bioterrorism agents. BoNT/G is the least studied of the seven serotypes. In an effort to further characterize the holotoxin and neurotoxin-associated proteins (NAPs), we conducted an <it>in silico </it>and proteomic analysis of commercial BoNT/G complex. We describe the relative quantification of the proteins present in the/G complex and confirm our ability to detect the toxin activity <it>in vitro</it>. In addition, we review previous literature to provide a complete description of the BoNT/G complex.</p> <p>Results</p> <p>An in-depth comparison of protein sequences indicated that BoNT/G shares the most sequence similarity with the/B serotype. A temperature-modified Endopep-MS activity assay was successful in the detection of BoNT/G activity. Gel electrophoresis and in gel digestions, followed by MS/MS analysis of/G complex, revealed the presence of four proteins in the complexes: neurotoxin (BoNT) and three NAPs--nontoxic-nonhemagglutinin (NTNH) and two hemagglutinins (HA70 and HA17). Rapid high-temperature in-solution tryptic digestions, coupled with MS/MS analysis, generated higher than previously reported sequence coverages for all proteins associated with the complex: BoNT 66%, NTNH 57%, HA70 91%, and HA17 99%. Label-free relative quantification determined that the complex contains 30% BoNT, 38% NTNH, 28% HA70, and 4% HA17 by weight comparison and 17% BoNT, 23% NTNH, 42% HA70, and 17% HA17 by molecular comparison.</p> <p>Conclusions</p> <p>The <it>in silico </it>protein sequence comparisons established that the/G complex is phenetically related to the other six serotypes of <it>C. botulinum</it>. Proteomic analyses and Endopep-MS confirmed the presence of BoNT and NAPs, along with the activity of the commercial/G complex. The use of data-independent MS^Edata analysis, coupled to label-free quantification software, suggested that the weight ratio BoNT:NAPs is 1:3, whereas the molar ratio of BoNT:NTNH:HA70:HA17 is 1:1:2:1, within the BoNT/G progenitor toxin.</p

The oscillation of mitotic kinase governs cell cycle latches in mammalian cells

The mammalian cell cycle alternates between two phases: S-G2-M with high levels of A- and B-type cyclin-dependent kinases (CycA,B:CDK); and G1 with persistent degradation of CycA,B by Cdh1-activated APC/C (anaphase promoting complex/cyclosome). Because CDKs phosphorylate and inactivate Cdh1, these two phases are mutually exclusive. This 'toggle switch' is flipped from G1 to S by cyclin-E (CycE:CDK), which is not degraded by Cdh1:APC/C; and from M to G1 by Cdc20:APC/C, which is not inactivated by CycA,B:CDK. After flipping the switch, cyclin E is degraded and Cdc20:APC/C is inactivated. Combining mathematical modelling with single-cell timelapse imaging, we show that dysregulation of CycB:CDK disrupts strict alternation of the G1-S and M-G1 switches. Inhibition of CycB:CDK results in Cdc20-independent Cdh1 'endocycles', and sustained activity of CycB:CDK drives Cdh1-independent Cdc20 endocycles. Our model provides a mechanistic explanation for how whole genome doubling can arise, a common event in tumorigenesis that can drive tumour evolution

Association between socioeconomic status, sex, and age at death from cystic fibrosis in England and Wales (1959 to 2008): cross sectional study

Objective To determine the trend in the association between socioeconomic status and sex and median age at death from cystic fibrosis in England and Wales, over the past 50 years

Apert syndrome with frontonasal encephalocele

We describe a female infant with Apert syndrome (acrocephalosyndactyly, type I) and a frontonasal encephalocele with unremarkable family history.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/38238/1/1320210422_ftp.pd

The relative influence of demographic, individual, social, and environmental factors on physical activity among boys and girls

<p>Abstract</p> <p>Background</p> <p>This study aimed to evaluate the associations of selected demographic, individual, social, and environmental factors with moderate-to-vigorous physical activity (MVPA) in a sample of children and adolescents.</p> <p>Methods</p> <p>MVPA was assessed among youth (n = 294) 10-17-years-old using the ActiGraph accelerometer. Youth completed measures of demographic and individual variables related to physical activity (PA), perceived social support by parents and peers, and perceived neighborhood characteristics. Parents completed the long-form of the International Physical Activity Questionnaire. The Physical Activity and Media Inventory was used to measure the home environment and Geographical Information Systems software was used to measure the physical neighborhood environment. Bivariate correlations and hierarchical multiple regression were conducted stratified by gender.</p> <p>Results</p> <p>Boys participated in significantly more MVPA than girls. In hierarchical analyses, peer support, home PA equipment, and temperature were significantly associated with MVPA among boys whereas distance to the school they attended was associated with MVPA among girls. The final models accounted for 25% and 15% of the variance in MVPA among boys and girls, respectively.</p> <p>Conclusions</p> <p>Important differences exist among the individual, social, and environmental factors related to MVPA between boys and girls. Boys' levels of activity appear to be influenced by factors closely linked to unstructured and social types of activities whereas girls' activities relate to internal and external barriers as well as their proximity to their schools. The prospective contribution of these important individual, social, and environmental factors to changes in MVPA among children and adolescents remains to be determined.</p

Polarization and relaxation of radon

Investigations of the polarization and relaxation of $^{209}$Rn by spin exchange with laser optically pumped rubidium are reported. On the order of one million atoms per shot were collected in coated and uncoated glass cells. Gamma-ray anisotropies were measured as a signal of the alignment (second order moment of the polarization) resulting from the combination of polarization and quadrupole relaxation at the cell walls. The temperature dependence over the range 130$^\circ$C to 220$^\circ$C shows the anisotropies increasing with increasing temperature as the ratio of the spin exchange polarization rate to the wall relaxation rate increases faster than the rubidium polarization decreases. Polarization relaxation rates for coated and uncoated cells are presented. In addition, improved limits on the multipole mixing ratios of some of the main gamma-ray transitions have been extracted. These results are promising for electric dipole moment measurements of octupole-deformed $^{223}$Rn and other isotopes, provided sufficient quantities of the rare isotopes can be produced.Comment: 4 pages, 4 figure