Platelet-related properties of the four diastereomers of compound <b>1</b>.

a<p>VASP, vasodilator-stimulated phosphoprotein.</p>b<p>All comparisons <i>vs.</i> diastereomer <b>1.1</b>.</p>c<p>p<0.05.</p>d<p>p<0.01.</p>e<p>p<0.001.</p

Chemical structure and stereo-configuration of the stereoisomers of diadenosine 5′,5″″-P¹,P⁴-dithio-P²,P³-chloromethylenetetraphosphate (compound 1), and of adenosine 5′-(P¹-thio-P²,P³-chloromethylenetriphosphate), (compound 2).

<p>R_P and S_P designate the absolute configuration of chiral P¹- and P⁴-phosphorothioates; r and s, the absolute configuration of the pseudo-asymmetric carbon of the P²,P³-chloromethylene group in compound <b>1</b>; R and S, the absolute configuration of the chloromethylene group in compound <b>2</b>. Ade, 5′-adenosyl; N.A., Not Asymmetric.</p

³¹P NMR Spectra (¹H decoupled) of the diastereomers of compound 1 and their racemic mixture.

<p>In the left panel are the P¹,P⁴ regions, and in the right panel are the P²,P³ regions of the spectra. The traces are, from bottom to top: racemic mixture (RM), diastereomer <b>1.1</b>, <b>1.2</b>, <b>1.3</b>, and <b>1.4</b>.</p

Effects of compound 1 diastereomers on platelet aggregation and platelet ADP receptor signaling.

<p>Inhibition by the diastereomers of compound <b>1</b> of: A, 3 µM ADP-stimulated platelet aggregation; B, P2Y₁₂ mediated ADP-induced decrease of VASP phosphorylation; and C, P2Y₁-mediated ADP-induced intraplatelet Ca²⁺ level increase. VASP, vasodilator-stimulated phosphoprotein. For other abbreviation, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094780#pone-0094780-g001" target="_blank">Figure 1</a>.</p

Reversed-phase HPLC separation of the diastereomers of compound 1.

<p>Panel A: chromatogram of the racemic mix of the diastereomers of compound <b>1</b> analyzed by reversed-phase HPLC using a XBridge C18 column, 3.5 µm, 4.6×150 mm (Waters Inc., Waltham, Mass. USA), linear gradient from 1 to 7% methanol in 20 mM potassium phosphate buffer, pH 7, 1 ml/min, detection by UV at 260 nm; Panel B: chromatograms of isolated individual diastereomers (<b>1.1</b>–<b>1.4</b>) of compound <b>1</b> analyzed using the same column and flow rate, but with isocratic elution with 7% methanol in 20 mM potassium phosphate buffer, pH 7.</p

Stability and metabolism of the four diastereomers of compound 1 in human plasma at 37 °C.

<p>t_1/2, half-life of first order elimination. For other abbreviations, see Figure 1.</p

Additional file 1: of No difference in the functional improvements between unilateral and bilateral total knee replacements

Table S1. Characteristics between patients who loss to follow-up or not. (DOCX 35 kb

Angiotensin-Converting Enzyme Insertion/Deletion Polymorphism and Susceptibility to Osteoarthritis of the Knee: A Case-Control Study and Meta-Analysis

<div><p>Background</p><p>Studies of angiotensin-converting enzyme insertion/deletion (ACE I/D) polymorphisms and the risks of knee osteoarthritis (OA) have yielded conflicting results.</p><p>Objective</p><p>To determine the association between ACE I/D and knee OA, we conducted a combined case-control study and meta-analysis.</p><p>Methods</p><p>For the case-control study, 447 knee OA cases and 423 healthy controls were recruited between March 2010 and July 2011. Knee OA cases were defined using the Kellgren-Lawrence grading system, and the ACE I/D genotype was determined using a standard polymerase chain reaction. The association between ACE I/D and knee OA was detected using allele, genotype, dominant, and recessive models. For the meta-analysis, PubMed and Embase databases were systematically searched for prospective observational studies published up until August 2015. Studies of ACE I/D and knee OA with sufficient data were selected. Pooled results were expressed as odds ratios (ORs) with corresponding 95% confidence intervals (CI) for the D versus I allele with regard to knee OA risk.</p><p>Results</p><p>We found no significant association between the D allele and knee OA [OR: 1.09 (95% CI: 0.76–1.89)] in the present case-control study, and the results of other genetic models were also nonsignificant. Five current studies were included, and there were a total of six study populations after including our case-control study (1165 cases and 1029 controls). In the meta-analysis, the allele model also yielded nonsignificant results [OR: 1.37 (95% CI: 0.95–1.99)] and a high heterogeneity (I²: 87.2%).</p><p>Conclusions</p><p>The association between ACE I/D and knee OA tended to yield negative results. High heterogeneity suggests a complex, multifactorial mechanism, and an epistasis analysis of ACE I/D and knee OA should therefore be conducted.</p></div

Selected results from the meta-analysis of angiotensin-converting enzyme insertion/deletion (ACE I/D) and knee osteoarthritis (OA).

<p>The top left subplot is a forest plot based on an allele model assumption (reference: I allele), and the top right subplot is a funnel plot based on the allele model assumption. The allele model is the most common method for detecting gene–disease associations; however, we found no significant signal in the allele model. However, the funnel plot indicates good symmetry in this meta-analysis. Results obtained with the dominant and recessive models are presented at the bottom. All results were nonsignificant.</p

Odds ratios of angiotensin-converting enzyme insertion/deletion (I/D) and knee osteoarthritis using assumptions from allele type, genotype, dominant, and recessive models.

<p>Odds ratios of angiotensin-converting enzyme insertion/deletion (I/D) and knee osteoarthritis using assumptions from allele type, genotype, dominant, and recessive models.</p