7-Selenabicyclo2.2.1heptane

Thermolysis of a benzene solution of N-4-(p-(methoxybenzyl)seleno) cyclohexanoyl-N,S-dimethyldithiocarbonate affords the hitherto unknown 7-selenabicyclo2.2.1heptane in 48% conversion and in 20% yield after chromatography. G3(MP2)-RAD calculations predict a rate constant of 5 X 104 s-1 at 80 °C (3.8 X 106 s -1 at 200 °C) for the intramolecular homolytic substitution process involved in this cyclization

Intramolecular homolytic substitution in selenoxides and selenones

G3(MP2)-RAD calculations provide activation energies for intramolecular homolytic substitution in the 4-(alkylselenoxo)butyl and 4-(alkylselendioxo)butyl radicals ranging from 21–39 kJ mol−1, and 143–170 kJ mol−1 for the selenoxide and selenone, respectively. Arrhenius data translate into rate constants for ring-closure of 1.5×105−2.5×108 s−1 (80°) for the selenoxides, and 5.4×10−14−5.1×10−11 s−1 (80°) for the corresponding selenones. NBO analyses show alkyl radicals are electrophilic during homolytic substitution at selenoxide selenium. The dominant orbital interaction in the transition state is worth 2413 kJ mol−1 and involves the SOMO and the lone-pair of electrons on selenium. The corresponding selenones are calculated to ring-close through transition states in which alkyl radicals are nucleophilic, but involve weak (SOMO--> σ* and SOMO--> π*) interactions. Consequently, this chemistry is not viable for selenones because of the lack of lone-pairs of electrons on the chalcogen

Understanding vaccine hesitancy in Canada: Results of a consultation study by the Canadian Immunization Research Network

"Vaccine hesitancy" is a concept now frequently used in vaccination discourse. The increased popularity of this concept in both academic and public health circles is challenging previously held perspectives that individual vaccination attitudes and behaviours are a simple dichotomy of accept or reject. A consultation study was designed to assess the opinions of experts and health professionals concerning the definition, scope, and causes of vaccine hesitancy in Canada. We sent online surveys to two panels (1- vaccination experts and 2- front-line vaccine providers). Two questionnaires were completed by each panel, with data from the first questionnaire informing the development of questions for the second. Our participants defined vaccine hesitancy as an attitude (doubts, concerns) as well as a behaviour (refusing some / many vaccines, delaying vaccination). Our findings also indicate that both vaccine experts and front-line vaccine providers have the perception that vaccine rates have been declining and consider vaccine hesitancy an important issue to address in Canada. Diffusion of negative information online and lack of knowledge about vaccines were identified as the key causes of vaccine hesitancy by the participants. A common understanding of vaccine hesitancy among researchers, public health experts, policy-makers and health care providers will better guide interventions that can more effectively address vaccine hesitancy within Canada

PC1, PC2, EGFR, OFD1, and flotillin-2 are part of a multimeric protein complex in RCTE and MO6-G3 cells.

<p>RCTE and MO6-G3 cells were grown 5 days post-confluency to allow for polarization and ciliogenesis. PC1 was immunoprecipitated using NM002 pAb and precipitated proteins were separated by SDS-PAGE and probed for PC1 using NM005 pAb (A, B), PC2 using AbCam pAb (C), EGFR using GeneTex pAb (D), OFD1 using Novus Biologicals pAb (E) and flotillin-2 using Cell Signaling Rabbit mAb (F). Bar graph showing a densitometric quantification of PC1 co-immunoprecipitation for PC1 (a, b), PC2 (c), EGFR (d), OFD1 (e), and flotillin-2 (f). Normal rabbit IgG was used as a negative control. In a reciprocal experiment EGFR was immunoprecipitated using Santa Cruz pAb from RCTE and MO6-G3 cell lysates. Immunoprecipitated proteins were separated by SDS-PAGE and probed for EGFR using GeneTex pAb (G), PC1 using NM002 pAb (H), PC2 using Santa Cruz pAb (I), OFD1 using Novus Biologicals pAb (J) and flotillin-2 using Cell Signaling Rabbit mAb (K). Bar graph showing a densitometric quantification of EGFR co-immunoprecipitation probed for EGFR (g), PC1 (h), PC2 (i), OFD1 (j), and flotillin-2 (k). Normal rabbit IgG was used as a negative control. Lysate lane inputs were 5% of immunoprecipitations. 25 µg of total protein was loaded into each well. Arrows indicate the quantified band of interest in each immunoblot panel. Bar represents the mean ± SD of at least three independent experiments. (*) p = 0.01 to 0.05, (**) p = 0.001 to 0.01, (***) p<0.001.</p

Key ciliary signaling proteins are expressed in RCTE and MO6-G3 cells.

<p>Renal cortical tubular epithelial (RCTE) cells and tooth derived odontoblasts (MO6-G3) cells were lysed and probed with antibodies directed against indicated proteins. PC1 using NM002 pAb (A, B), PC2 using Santa Cruz pAb (C), OFD1 using Santa Cruz pAb <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0106330#pone.0106330-Romio1" target="_blank">[3]</a> (D), EGFR using Santa Cruz pAb (E), ErbB2 using US Biological pAb (F), flotillin-1 using BD Transduction mAb (H), and flotillin-2 using BD Transduction mAb (I) were found to be expressed in both cell types. Actin mAb from Millipore (G) was used as a loading control for PC1, PC2, EGFR, ErbB2, and OFD1; α/β-tubulin pAb from Cell Signaling (J) was used as a loading control for flotillin-1 and flotillin-2. Actin or α/β-tubulin was used to normalize results for quantification. Bar graph showing densitometric quantification of PC1 (a, b), PC2 (c), OFD1 (d), EGFR (e), ErbB2 (f), flotillin-1 (g), flotillin-2 (h). Bar graph represents the mean ± SD of four independent experiments. (*) p = 0.01 to 0.05, (***) p<0.001.</p

Endogenous OFD1, PC1, PC2, EGFR and flotillin-1 localize to primary cilia of RCTE and MO6-G3 cells.

<p>Polarized RCTE and MO6-G3 cells were fixed and stained using the indicated antibodies. OFD1 from Novus Biologicals (A, F), PC1 using pAb NM002 (B, G), PC2 using AbCam pAb (C, H), EGFR using GeneTex pAb (D, I) and flotillin-1 using AbCam pAb (E, J) were found localized to 100% of primary cilia analyzed for both cell types. Acetylated α-tubulin (Sigma-Aldrich mAb) labeling identifies cilia. Zeiss LSM510 confocal microscope images (63× objective). Arrows denote the protein of interest within a cilium. Representative results from at least 5 independent experiments. Secondary antibody only controls were negative (not shown). Scale bar 10 µm.</p

Expression of PC1, OFD1, EGFR, and Flotillin-1 is decreased in ciliary fractions of PKD cells compared to RCTE cells.

<p>RCTE cells and PKD Q4004X cells were grown 6 days post-confluency to allow for cell polarization and ciliogenesis and treated with 1.5 mM Dibucaine-HCl for 5 minutes to reduce cell loss and induce shedding of primary cilia. Cilia were collected by fractionation and the ciliary fraction was probed. PC1 (MN032 pAb), OFD1 (Novus Biologicals pAb), EGFR (GeneTex pAb), and flotillin-1 (BD Transduction mAb) were expressed in the ciliary fraction of deciliated RCTE cells but not PKD cells. α/β-tubulin (Cell Signaling pAb) was used as a marker of primary cilia. The nuclear marker lamin B (Santa Cruz pAb) was not present in the ciliary fraction (A). Quantification of 3 independent samples (B). Mean values were found to be statistically different (p<0.0001) using 1-way ANOVA.</p

Key ciliary signaling proteins are significantly reduced in primary cilia of PKD cells.

<p>Human RCTE and PKD Q4004X cells (with expression of mutant PC1) were grown on coverslips 5 days post-confluency to permit ciliogenesis. Cells were fixed and stained using antibodies directed against indicated proteins: PC1 (NM002 pAb); PC2 (AbCam pAb); EGFR (GeneTex pAb); and flotillin-1 (AbCam pAb). PC1 (A), PC2 (C), EGFR (E) and flotillin-1(G) are present in primary cilia of RCTE cells. PC1 (B), PC2 (D), EGFR (F) and flotillin-1 (H) were lacking in primary cilia of PKD Q4004X cells. Acetylated α-tubulin (Sigma-Aldrich mAb) staining identifies cilia. Arrows denote a small residual pool of EGFR detectable at the ciliary base of PKD cells. Zeiss LSM510 confocal microscope images (63× objective). Representative results from at least 5 independent experiments. Comparative images are from a single experiment and taken under identical settings. Arrows denote cilia. Scale bar 10 µm. Quantification shows individual ciliary protein intensities normalized to the respective ciliary volume (I–L). Each protein was quantified in 30–100 cilia for each cell type. z-stack images were imported into SlideBook and a volume mask for each cilium was created based on acetylated α-tubulin staining. Staining intensities for each protein were quantified within the respective ciliary volume mask. Statistical evaluation based on two-tailed t-test. (*) p = 0.0105, (***) p<0.0001.</p