Image of the MSCs and VECs on the PLGA surface from scanning electron microscopy

<p><b>Copyright information:</b></p><p>Taken from "In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds"</p><p>http://www.biomedical-engineering-online.com/content/6/1/41</p><p>BioMedical Engineering OnLine 2007;6():41-41.</p><p>Published online 4 Nov 2007</p><p>PMCID:PMC2186340.</p><p></p> A. MSCs adhere and extend on the PLGA surface. The arrow indicates the pseudopodium (× 1000). B. VECs adhere on the PLGA surface as an oval and stone-like shape (× 1000)

Soft X-ray images from right and left thigh and X-ray density analysis using NIH image software

<p><b>Copyright information:</b></p><p>Taken from "In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds"</p><p>http://www.biomedical-engineering-online.com/content/6/1/41</p><p>BioMedical Engineering OnLine 2007;6():41-41.</p><p>Published online 4 Nov 2007</p><p>PMCID:PMC2186340.</p><p></p> A. Right thigh implanted with the PLGA seeded with the MSCs and VECs. B. Left thigh implanted with the PLGA seeded with the MSCs. C. X-ray density analysis. The data shown are the means ± SD from six experiments

ALP activity and osteocalcin synthesis from three-dimensional co-culture in vitro

<p><b>Copyright information:</b></p><p>Taken from "In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds"</p><p>http://www.biomedical-engineering-online.com/content/6/1/41</p><p>BioMedical Engineering OnLine 2007;6():41-41.</p><p>Published online 4 Nov 2007</p><p>PMCID:PMC2186340.</p><p></p> A. The ALP activity. B. Osteocalcin synthesis. The data shown are the means ± SD from six experiments. (I) means indirect. (D) means direct

Hematoxylin and eosin staining of the implanted area from rat thigh and Quantitative measurements of new blood vessel and new bone from HE staining slides

<p><b>Copyright information:</b></p><p>Taken from "In vitro and in vivo effects of rat kidney vascular endothelial cells on osteogenesis of rat bone marrow mesenchymal stem cells growing on polylactide-glycoli acid (PLGA) scaffolds"</p><p>http://www.biomedical-engineering-online.com/content/6/1/41</p><p>BioMedical Engineering OnLine 2007;6():41-41.</p><p>Published online 4 Nov 2007</p><p>PMCID:PMC2186340.</p><p></p> A. Control group at 8 weeks post-implanted. B. Experiment group at 8 weeks post-implanted. C. Control group at 12 weeks post-implanted. D. Experiment group at 12 weeks post-implanted. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. Red arrow indicates PLGA material. Black arrow indicates new bone. Yellow arrow indicates blood vessel. E. New blood vessel. F. New bone. Control is left thigh implanted with the PLGA seeded with the MSCs. Experiment is right thigh implanted with the PLGA seeded with the MSCs and VECs. The data shown are the means ± SD from six experiments

Table_1_Higher blood cotinine level is associated with worse cognitive functioning in non-smoking older adults.DOCX

IntroductionSecondhand smoke (SHS) is common in older adults; however, its cognitive effect is unclear. We aimed to examine the association between serum cotinine level and cognitive functioning among non-smoking older adults.Materials and methodsA total of 2,703 older adults aged 60 and above from the National Health and Nutrition Examination (NHANES) Survey 2011–2014 were included. Serum cotinine level was analyzed in the laboratory. A level ≤10 ng/ml and a response of “no” to the question “Do you currently smoke?” were used to select non-smokers. Cognitive functioning was measured using the Consortium to Establish a Registry for Alzheimer’s disease Word Learning subtest (CERAD-WL) immediate and delayed recall tests, the Animal Fluency test (AFT), and the Digit Symbol Substitution test (DSST). Multivariable linear regression models were constructed to examine the association between serum cotinine level quartile and test-specific and global cognition z scores adjusting for age, race/ethnicity, education, depressive symptoms, body mass index, alcohol use, smoking history, prevalent coronary heart disease (CHD), stroke, and systolic blood pressure.ResultsAbout half of the participants (mean age 70.5 years) were female (53.6%), non-Hispanic White (48.3%), and completed some college and above (50.2%). Multivariate linear regressions with a reference group being those in the 1st quantile (lowest) showed that participants in the 4th quartile (highest) of serum cotinine level had lower immediate recall [β = −0.16, 95% confidence interval (CI) = −0.29, −0.03], AFT (β = −0.19, 95% CI = −0.33, −0.05), DSST (β = −0.27, 95% CI = −0.39, −0.15), and global cognition (β = −0.26, 95% CI = −0.39, −0.14) z scores. Participants in the 3rd quartile had lower immediate recall (β = −0.16, 95% CI = −0.30, −0.02) and global cognition (β = −0.16, 95% CI = −0.29, −0.02) z scores. Participants in the 2nd quartile had lower delayed recall z scores (β = −0.16, 95% CI = −0.29, −0.02).ConclusionHigher serum cotinine level was associated with worse cognitive functioning in non-smoking older adults. Prevention and reduction of SHS in older adults may help protect their cognitive functioning.</p

T_tridentatus_genome_contigs.fa

T_tridentatus genome contigs assembled as described in accompanying manuscript. NB .fasta.gz file, needs to be unpacked using gzip or similar before us

C_rotundicauda_chelicerate_transcriptome

C_rotundicauda chelicerate transcriptome assembled using Trinity as described in accompanying manuscript. .fasta.gz file, needs unpacking using gzip or similar before us

L_polyphemus_genome_contigs.fa

L polyphemus genome contigs assembled as described in accompanying manuscript. NB .fasta.gz file, needs to be unpacked using gzip or similar before us

T_tridentatus_chelicerate_transcriptome

T. tridentatus chelicerate transcriptome assembled using Trinity as described in accompanying manuscript. .fasta.gz file, needs unzipping using gzip or similar before use

Proteomic Quantification and Site-Mapping of <i>S</i>‑Nitrosylated Proteins Using Isobaric iodoTMT Reagents

<i>S</i>-Nitrosylation is a redox-based protein post-translational modification in response to nitric oxide signaling and is involved in a wide range of biological processes. Detection and quantification of protein <i>S</i>-nitrosylation have been challenging tasks due to instability and low abundance of the modification. Many studies have used mass spectrometry (MS)-based methods with different thiol-reactive reagents to label and identify proteins with <i>S</i>-nitrosylated cysteine (SNO-Cys). In this study, we developed a novel iodoTMT switch assay (ISA) using an isobaric set of thiol-reactive iodoTMTsixplex reagents to specifically detect and quantify protein <i>S</i>-nitrosylation. Irreversible labeling of SNO-Cys with the iodoTMTsixplex reagents enables immune-affinity detection of <i>S</i>-nitrosylated proteins, enrichment of iodoTMT-labeled peptides by anti-TMT resin, and importantly, unambiguous modification site-mapping and multiplex quantification by liquid chromatography–tandem MS. Additionally, we significantly improved anti-TMT peptide enrichment efficiency by competitive elution. Using ISA, we identified a set of SNO-Cys sites responding to lipopolysaccharide (LPS) stimulation in murine BV-2 microglial cells and revealed effects of <i>S</i>-allyl cysteine from garlic on LPS-induced protein <i>S</i>-nitrosylation in antioxidative signaling and mitochondrial metabolic pathways. ISA proved to be an effective proteomic approach for quantitative analysis of <i>S</i>-nitrosylation in complex samples and will facilitate the elucidation of molecular mechanisms of nitrosative stress in disease