Quantum Chemistry Calculations on the Mechanism of Isoquinoline Ring-Opening and Denitrogenation in Supercritical Water

Computational studies at the M06/6-311G­(d,p) and M06-2<i>X</i>/6-311+G­(d,p) levels were performed to explore the detailed mechanism of isoquinoline ring-opening and denitrogenation in a supercritical water system. Three reaction paths with the same product, 2-(2-oxoethyl) benzaldehyde, were supported by the computational results. The rate-limiting step in the major degradation reaction is an addition reaction at the N position. H₂O is added to both the 1C–2N double bond (1C–2N addition reaction) and the 2N–3C double bond (2N–3C addition reaction) of the isoquinoline molecule, where the oxygen of H₂O is added to the carbon atom. The energy barrier of the 1C–2N addition reaction is 52.7 kcal/mol, while that of 2N–3C addition (from Path 6) is 60.1 kcal/mol. From catalysis by two water molecules, the barrier of 1C–2N addition (Reaction (1)) is reduced to 27.5 kcal/mol. Catalysis from water molecule clusters is shown to considerably affect the process of isoquinoline ring-opening and denitrogenation, as indicated by comparing the reaction energy barrier heights with and without water catalysts

Additional file 1 of Sacubitril/valsartan inhibits the proliferation of vascular smooth muscle cells through notch signaling and ERK1/2 pathway

Supplementary Material

The Difference in Prognosis between Renal Sinus Fat and Perinephric Fat Invasion for pT3a Renal Cell Carcinoma: A Meta-Analysis

<div><p>Background</p><p>In the current Tumour-Node-Metastasis (TNM) classification system for renal cell carcinoma (RCC), both renal sinus fat invasion (SFI) and perinephric fat invasion (PFI) are defined as T3a, suggesting that the prognosis should be similar for the two pathologic findings. Several studies, however, have reported a worse prognosis for SFI in patients with a T3a tumor. In order to compare the prognosis of these two pathologic findings (SFI versus. PFI) in a more comprehensive way, this meta-analysis was performed.</p><p>Methods</p><p>To identify relevant studies, Medline, Embase, Cochrane Library, and Scopus database were searched from the inception until October 2014. A meta-analysis was performed using Review Manager 5.2 and STATA 11. Pooled Odds ratio (OR) and/or hazard ratio (HR) with 95% confidence interval (CI) were calculated to examine the risk or hazard association.</p><p>Results</p><p>A total of 6 studies including 1031 patients qualified for analysis. T3a RCC patients with SFI were significantly associated with poor cancer specific survival(CSS) (HR: 1.47, 95% CI: 1.19–1.83; P<0.001) compared to those with PFI. In T3aNx/N0M0 subgroup, SFI patients also showed a worse prognosis than those with PFI (CSS, HR: 1.94, 95% CI: 1.21–3.12; P = 0.006). T3a RCC patients with SFI had higher Furhman grade, greater possibility of lymph node metastasis, sarcomatoid differentiation and tumour necrosis. Main limitation is the relatively small number of included studies.</p><p>Conclusion</p><p>The present meta-analysis suggested that SFI is associated with worse CSS in patients with pT3a RCC. However, due to the small number of included studies, future studies with a large sample size are required to further verify our findings.</p></div

One-removed Analysis for CSS.

<p>One-removed Analysis for CSS.</p

Flow diagram of studies selection procedure.

<p>Flow diagram of studies selection procedure.</p

Meta-analysis of the association between SFI and clinicopathological parameters in overall T3a RCC.

<p>(A)Grade; (B) N status; (C) M status; (D) Sarcomatoid differentiation; (E) Tumour necrosis. N⁺: local lymph nodes positive; N⁻:local lymph nodes negative; M⁺: with distal metastasis; M⁻: without distal metastasis; SD⁺: with sarcomatoid differentiation; SD⁻: without sarcomatoid differentiation; CI: confidence interval; SE: standard error.</p

Optimized parameters for the 293GCAC3-based bioassay.

<p>Various parameters of the assay were optimized, including cell number, cell culture time, cGMP-HRP dilution rate and so on as listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049934#pone-0049934-t001" target="_blank">Table 1</a>.</p

Characteristics of the Included Studies.

<p>Characteristics of the Included Studies.</p

Funnel plot for all studies included in this meta-analysis.

<p>(A) Funnel plot assessing SFI and cancer specific survival (CSS) in T3a renal cell carcinoma patients. (B) Funnel plot assessing SFI and CSS in T3aNx/N0M0 patients. SE: standard error.</p

A Novel Bioassay for the Activity Determination of Therapeutic Human Brain Natriuretic Peptide (BNP)

<div><h3>Background</h3><p>Recombinant human brain natriuretic peptide (rhBNP) is an important peptide-based therapeutic drug indicated for the treatment of acute heart failure. Accurate determination of the potency of therapeutic rhBNP is crucial for the safety and efficacy of the drug. The current bioassay involves use of rabbit aortic strips, with experiments being complicated and time-consuming and markedly variable in results. Animal-less methods with better precision and accuracy should be explored. We have therefore developed an alternative cell-based assay, which relies on the ability of BNP to induce cGMP production in HEK293 cells expressing BNP receptor guanylyl cyclase-A.</p> <h3>Methodology/Principal Findings</h3><p>An alternative assay based on the measurement of BNP-induced cGMP production was developed. Specifically, the bioassay employs cells engineered to express BNP receptor guanylyl cyclase-A (GCA). Upon rhBNP stimulation, the levels of the second messager cGMP in these cells drastically increased and subsequently secreted into culture supernatants. The quantity of cGMP, which corresponds to the rhBNP activity, was determined using a competitive ELISA developed by us. Compared with the traditional assay, the novel cell-based assay demonstrated better reproducibility and precision.</p> <h3>Conclusion/Significance</h3><p>The optimized cell-based assay is much simpler, more rapid and precise compared with the traditional assay using animal tissues. To our knowledge, this is the first report on a novel and viable alternative assay for rhBNP potency analysis.</p> </div