Fluorescence and chemiluminescence properties of indolylmaleimides: experimental and theoretical studies

Ministry of Education, Culture, Sports, Science and Technology of Japan [17590034]Various indolylmaleimides (IMs) were synthesized, and their fluorescence (FL) and chemiluminescence (CL) were measured. The substitution at the 2-position of the indole ring and the 3- or 4-position of the maleimide moiety caused an obvious change in the FL and CL of the IMs. An almost on-off switching of the FL of the IMs was observed. The intramolecular charge transfer from the indole moiety to the maleimide moiety occurred in 3-(1H-3-indolyl)-2,5-dihydro-1H-2,5-pyrroledione. In the FL of the IMs, CASPT2 calculations showed deprotonation of the NH group of the indole ring and the maleimide moiety at the excited state. The C=C bond in the maleimide moiety was needed for strong CL in the IMs without substitution at the 2-position of the indole ring. The relationships between the FL or CL properties and the structures of the IMs were clarified. These results provide significant information on the rational design of IMs as FL and CL probes

Endothelin receptors and their cellular signal transduction mechanism in human cultured prostatic smooth muscle cells

1. Endothelin (ET) receptors, and their cellular signal transduction mechanism, were characterized in a primary culture of human prostatic smooth muscle cells (HP cell). 2. [(125)I]-ET-1 and [(125)I]-ET-3 binding studies revealed that both ET(A) and ET(B) receptors were present in the HP cells, and the ratio of ET(A) to ET(B) receptors was 1.4:1. 3. Analysis of ET receptor mRNA by reverse transcription-polymerase chain reaction also demonstrated that HP cells express both ET(A) and ET(B) receptors. 4. ET-1 and ET-3 increased intracellular free Ca(2+) concentration ([Ca(2+)](i)) in the HP cells in a concentration-dependent manner. Use of subtype selective antagonists BQ-123 and BQ-788, indicated that both ET(A) and ET(B) receptors were coupled to an increase in [Ca(2+)](i). 5. Pretreatment of the cells with pertussis toxin resulted in a significant but partial attenuation of the [Ca(2+)](i) increase mediated through the ET(A) and ET(B) receptors. However, sensitivity to pertussis toxin (PTX) was significantly different between them. 6. In conclusion, HP cells possess ET(A) and ET(B) receptors. Further, these two endothelin receptor subtypes evoke an increase in [Ca(2+)](i) possibly via the action of different GTP-binding proteins

Reference intervals of serum hyaluronic acid corresponding to the radiographic severity of knee osteoarthritis in women

<p>Abstract</p> <p>Backgroud</p> <p>While serum levels of hyarulonic acid (sHA) is known to be useful for a burden of disease biomarker in knee OA, it is far from practical. The reference intervals must be established for biomarkers to be useful for clinical interpretation. The aim of this study was to establish the reference intervals of sHA corresponding to the radiographic severity of knee OA for elucidating whether sHA can be useful as a burden of disease marker for individual patient with knee OA.</p> <p>Methods</p> <p>372 women with Kellgren & Lawrence grade (K/L) 1 through 4 painful knee OA were enrolled in this study. The patients included 54 with K/L 1, 96 with K/L 2, 97 with K/L 3, and 118 with K/L 4. Serum samples were obtained from all subjects on the day that radiographs taken. A HA binding protein based latex agglutination assay that employed an ELISA format was used to measure sHA. Age and BMI adjusted one way ANOVA was used to set the reference intervals of sHA.</p> <p>Results</p> <p>The reference intervals for sHA corresponding to the patients with K/L 4 (49.6 – 66.5 ng/ml) was established without any overlap against to those with K/L 1, 2 and 3, while those with K/L 1, 2 and 3 showed considerable overlap.</p> <p>Conclusions</p> <p>These results indicate that sHA can be available as a burden of disease marker for the individuals with severe knee OA (K/L 4), while it is not for those with primary to moderate knee OA (K/L 1–3).</p