Correlation of Solubility and Prediction of the Mixing Properties of Capsaicin in Different Pure Solvents

Using a static analytical model, experimental solubility data were obtained for capsaicin in <i>n</i>-hexane, cyclohexane, carbon disulfide, butyl ether, and isopropyl ether at temperatures ranging from 278.15 to 323.15 K. The melting temperature and fusion enthalpy of capsaicin were measured using differential scanning calorimetry. The measured solubility data were well correlated by the van’t Hoff, modified Apelblat, λ<i>h</i> (Buchowski), Wilson, and NRTL models, with the Wilson model showing the best agreement. The activity coefficients of capsaicin and the mixing Gibbs free energies, enthalpies, and entropies of the resulting solutions were predicted on the basis of the Wilson model parameters at measured solubility points. In addition, the infinite-dilution activity coefficients and excess enthalpies of capsaicin were estimated. Finally, the effects of solute–solvent intermolecular repulsive interactions on the solubility behavior and the values of mixing Gibbs free energy were discussed

The NRF-1 binding site in the <i>IDE</i> promoter is conserved among different species.

<p>(<b>A</b>) The NRF-1 binding site in the <i>IDE</i> promoter is conserved among different species. The underlined region indicates the conserved NRF-1 binding motif. (<b>B</b>) The NRF-1 binding motif is critical for human <i>IDE</i> transcription initiation. Different truncations of the human <i>IDE</i> promoter were cloned into the pGL3-basic vector. Luciferase activity of the reporter plasmids in HeLa cells is represented as fold of the pGL3-basic vector. (<b>C</b>) ChIP. NRF-1 binding to the <i>IDE</i> promoter in HeLa cells was determined by ChIP. The promoter of <i>cytochrome c</i> (<i>cyt c</i>) is used as a positive control for NRF-1 binding, while the promoter of <i>GAPDH</i> acts as a negative control. (<b>D</b>) The NRF-1 binding motif is essential for the effect of dominant negative NRF-1 on human <i>IDE</i> promoter activity. HeLa cells were transiently co-transfected with wild-type (−484/+173) or NRF-1 binding site-mutated (NRF-mut) human <i>IDE</i> reporter plasmids (0.4 µg) and <i>Renilla</i> luciferase plasmid (4 ng) along with or without dominant negative (DN) NRF-1 expression plasmids (0.4 µg). Twenty-four hours after transfection, cells were lysed, and the luciferase activity was examined. Firefly luminescence signal was normalized based on the <i>Renilla</i> luminescence signal.</p

The mouse <i>IDE</i> promoter contains a CpG island and has dispersed transcription initiation sites.

<p>(<b>A</b>) Representation of the mouse <i>IDE</i> promoter. The dashed line indicates the unknown region which was cloned and sequenced in this study. The mouse <i>IDE</i> promoter contains a CpG island with a length of approximately 1300 bp. (<b>B</b>) The transcription initiation sites of mouse <i>IDE</i>. The frequency of transcription initiation from different sites is shown. The mouse <i>IDE</i> promoter has dispersed transcription initiation sites located within a window of 62 bp. The first transcription initiation site is underlined.</p

Nuclear Respiratory Factor 1 Mediates the Transcription Initiation of <em>Insulin-Degrading Enzyme</em> in a TATA Box-Binding Protein-Independent Manner

<div><p>CpG island promoters often lack canonical core promoter elements such as the TATA box, and have dispersed transcription initiation sites. Despite the prevalence of CpG islands associated with mammalian genes, the mechanism of transcription initiation from CpG island promoters remains to be clarified. Here we investigate the mechanism of transcription initiation of the CpG island-associated gene, <em>insulin-degrading enzyme</em> (<em>IDE</em>). <em>IDE</em> is ubiquitously expressed, and has dispersed transcription initiation sites. The <em>IDE</em> core promoter locates within a 32-bp region, which contains three CGGCG repeats and a nuclear respiratory factor 1 (NRF-1) binding motif. Sequential mutation analysis indicates that the NRF-1 binding motif is critical for <em>IDE</em> transcription initiation. The NRF-1 binding motif is functional, because NRF-1 binds to this motif <em>in vivo</em> and this motif is required for the regulation of <em>IDE</em> promoter activity by NRF-1. Furthermore, the NRF-1 binding site in the <em>IDE</em> promoter is conserved among different species, and dominant negative NRF-1 represses endogenous IDE expression. Finally, TATA-box binding protein (TBP) is not associated with the <em>IDE</em> promoter, and inactivation of TBP does not abolish <em>IDE</em> transcription, suggesting that TBP is not essential for <em>IDE</em> transcription initiation. Our studies indicate that NRF-1 mediates <em>IDE</em> transcription initiation in a TBP-independent manner, and provide insights into the potential mechanism of transcription initiation for other CpG island-associated genes.</p> </div

The NRF-1 binding motif in the mouse <i>IDE</i> promoter is functional.

<p>(<b>A</b>) Representation of wild-type (−136/+139 WT) and the NRF-1 binding site-mutated (NRF-mut) luciferase reporter plasmids of the mouse <i>IDE</i> promoter. (<b>B</b>) and (<b>C</b>) Dominant negative NRF-1 represses <i>IDE</i> promoter activity. NIH-3T3 and HeLa cells were transiently co-transfected with wild-type (-136/+139 WT) or NRF-1 binding site-mutated (NRF-mut) <i>IDE</i> reporter plasmids (0.4 µg) and <i>Renilla</i> luciferase plasmid (4 ng) along with or without dominant negative (DN) NRF-1 expression plasmids (0.4 µg). Twenty-four hours after transfection, cells were lysed, and the luciferase activity was examined. Firefly luminescence signal was normalized based on the <i>Renilla</i> luminescence signal. (<b>D</b>) ChIP. NRF-1 binding to the <i>IDE</i> promoter in NIH-3T3 cells was determined by ChIP. The promoter of <i>cytochrome c</i> (<i>cyt c</i>) is used as a positive control for NRF-1 binding, while the promoter of <i>GAPDH</i> acts as a negative control.</p

TBP is not required for <i>IDE</i> transcription initiation.

<p>(<b>A</b>) and (<b>B</b>) TBP is not associated with the <i>IDE</i> promoter. Binding of TBP and RNA polymerase II to the <i>IDE</i> promoter in NIH-3T3 (<b>A</b>) and HeLa (<b>B</b>) cells was tested by ChIP assays. Promoters of <i>GAPDH</i> and <i>EF1α1</i> were used as positive controls for TBP and RNA polymerase II binding. Negative controls were also included. Data are represented as relative enrichment to the input. (<b>C</b>) <i>In vitro</i> transcription assays. DNA template for the <i>CMV</i>, mouse <i>IDE</i> or human <i>IDE</i> promoter was incubated with HeLa nuclear extracts and ribonucleotides. The resulting transcripts were purified, digested with DNase I to eliminate the DNA template and detected by RT-PCR. An internal control RNA was included to indicate the purification efficiency of different samples. Complete elimination of the DNA template was confirmed by PCR. Transcription from all the three DNA templates proceeded only when both the DNA template and HeLa nuclear extracts existed, and was inhibited by 1 µg/mL of α-amanitin. (<b>D</b>) Heat-inactivation of TBP does not block <i>IDE</i> transcription. HeLa nuclear extracts were heated at 47°C for 15 min before <i>in vitro</i> transcription assays.</p

Mapping the core promoter region of mouse <i>IDE</i>.

<p>(<b>A</b>) and (<b>B</b>) The region between −23 and +9 of the mouse <i>IDE</i> promoter is essential for transcription initiation and behaves as the core promoter. Different truncations of the mouse <i>IDE</i> promoter were cloned into the pGL3-basic vector. NIH-3T3 or HeLa cells were transiently transfected with luciferase reporter plasmids of the mouse <i>IDE</i> promoter (0.8 µg) and <i>Renilla</i> luciferase reporter plasmid (pCMV-RL, 8 ng). Twenty-four hours after transfection, cells were lysed, and the luciferase activity was determined. Firefly luminescence signal was normalized based on the <i>Renilla</i> luminescence signal. Data are represented as fold of the firefly luciferase activity of the pGL3-basic vector.</p

Sequential mutation of the mouse <i>IDE</i> core promoter.

<p>The mouse <i>IDE</i> core promoter contains three CGGCG repeats and a NRF-1 binding motif. The core promoter region with sequential mutations (M1 to M11) was cloned into the pGL3-basic vector. Luciferase activity of the reporter plasmids in NIH-3T3 and HeLa cells is represented as a percentage of the wild-type (WT) reporter plasmid.</p

Solid–Liquid Phase Equilibrium and Mixing Properties of Cloxacillin Benzathine in Pure and Mixed Solvents

Experimental solubility data of cloxacillin benzathine in pure solvents and binary solvent mixtures from 278.15 to 313.15 K were measured using a multiple reactor setup. The measured data in pure solvents were correlated by the van’t Hoff equation, modified Apelblat equation, <i>λh</i> equation, Wilson model, and NRTL model, and the Wilson model showed the best agreement. Thus, the activity coefficients of cloxacillin benzathine as well as the mixing Gibbs free energies, enthalpies, and entropies of the solutions were predicted with the correlation of experimental data based on the Wilson model. Some other properties were also estimated, including the infinite-dilution activity coefficients and excess enthalpies in pure solvents. The solubility data in binary solvent mixtures as a function of solvent composition were correlated by the Wilson model. The negative values of the calculated partial molar Gibbs free energies indicated the variation trend of the solubility

DataSheet_1_Causal association between celiac disease and inflammatory bowel disease: A two-sample bidirectional Mendelian randomization study.doc

BackgroundAn epidemiological link between celiac disease (CeD) and inflammatory bowel disease (IBD) has been well established recently. In this study, Mendelian randomization (MR) analysis was performed employing pooled data of publicly available genome-wide association studies (GWAS) to determine the causal relationship between CeD and IBD, encompassing ulcerative colitis (UC) and Crohn’s disease (CD).MethodsDataset of CeD was acquired from GWAS for 12,041 cases and 12,228 controls. A GWAS of more than 86,000 patients and controls was used to identify genetic variations underlying IBD. MR analyses were performed with an inverse-variance-weighted approach, an MR-Egger regression, a weighted-mode approach, a weighted-median method, and sensitivity analyses of MR pleiotropy residual sum and outlie (MR-PRESSO).ResultsMR demonstrated that genetic predisposition to CeD was linked to a augmented risk of IBD (OR: 1.1408; 95% CI: 1.0614-1.2261; P = 0.0003). In the analysis of the two IBD subtypes, genetic predisposition to CeD was also linked to increased risks of UC (OR: 1.1646; 95% CI: 1.0614-1.2779; P = 0.0012) and CD (OR: 1.1865; 95% CI: 1.0948-1.2859; P = 3.07E-05). Reverse MR analysis results revealed that genetic susceptibility to IBD and CD was correlated with an augmented risk of CeD. However, there was no genetic correlation between UC and CeD. All of the above results were validated with other GWAS databases.ConclusionThere is a bidirectional causal relationship of CeD with IBD and CD. However, UC only augments the risk of developing CeD.</p