Pd-Catalyzed Debenzylation and Deallylation of Ethers and Esters with Sodium Hydride

Herein we demonstrate simply that the addition of Pd­(OAc)₂ as a promotor switches the reactivity of a commonly used base NaH to a nucleophilic reductant. The reactivity is engineered into a palladium-catalyzed reductive debenzylation and deallylation of aryl ethers and esters. This operationally simple, mild protocol displays a broad substrate scope and a broad spectrum of functional group tolerance (>50 examples) and high chemoselectivity toward aryl ethers over aliphatic structures. Moreover, the dual reactivity of NaH as a base and a reductant is demonstrated in efficient synthetic elaboration

Practical Prediction of Ten Common <i>Streptococcus pneumoniae</i> Serotypes/Serogroups in One PCR Reaction by Multiplex Ligation-Dependent Probe Amplification and Melting Curve (MLPA-MC) Assay in Shenzhen, China

<div><p>Background</p><p><i>Streptococcus pneumoniae</i> has more than 95 distinct serotypes described to date. However, only certain serotypes are more likely to cause pneumococcal diseases. Thus serotype surveillance is important for vaccine formula design as well as in post-vaccine serotype shift monitor. The goal of this study was to develop a practical screening assay for ten Shenzhen China common pneumococcal serotypes/serogroups in one molecular reaction.</p><p>Methods</p><p>A molecular assay, based on multiplex ligation-dependent probe amplification (MLPA) and melting curve (MC) analysis, was developed in an integrated approach (MLPA-MC) for the detection of ten capsular serotypes/serogroups 4, 6 (6A/6B/6C/6D), 9V/9A, 14, 15F/15A, 15B/15C, 18 (18F/18A/18B/18C), 19F, 19A and 23F. We designed serotype/serogroup-specific MLPA probes and fluorescent detection probes to discriminate the different serotypes/serogroups in one molecular reaction. The three steps of MLPA-MC assay are continuous reactions in one well detected by LightCycler 480. A total of 210 <i>S</i>. <i>pneumoniae</i> isolates from our local Maternity and Child Health Hospital were randomly chosen to evaluate the assay against published multiplex PCR assays.</p><p>Results</p><p>Our results showed that 198 (94.3%) of <i>S</i>. <i>pneumoniae</i> isolates were type-able by our assays and the results were in complete concordance with the published multiplex PCRs. Using the MLPA-MC assay, 96 <i>S</i>. <i>pneumoniae</i> isolates could be typed within 3 hours with limited hands-on time. This serotype/serogroup-screening assay can be easily modified or extended by modification of the serotype/serogroup-specific MLPA probes combinations according to the needs of different laboratories.</p><p>Conclusions</p><p>We recommend use of this assay as a starting point for screening serotype/serogroup frequencies. There is a need for this assay to be combined with other molecular typing assays, like published serotype specific PCRs, or even the Quellung reaction for serotype confirmation.</p></div

Overview of the MLPA-MC assay.

<p>(A) Hybridization: A pair of MLPA probe consists of left and right probe: (a) Universal forward primer binding site (5’-GGGTTCCCTAAGGGTTGGA-3’) will be the same to the universal forward primer (5’-GGGTTCCCTAAGGGTTGGA-3’), (b) Left probe oligonucleotides (LPO): Left serotype specific hybridization regions; (c) Right probe oligonucleotides (RPO): Right serotype specific hybridization regions, (d) Serotype specific stuffer sequence, (e) Universal reverse primer binding site (5’-TCTAGATTGGATCTTGCTGGCAC-3’) will be targeted by universal reverse primer, 5’-GTGCCAGCAAGATCCAAT-(labeled FAM)CTAGA-3’. The LPO and RPO hybridize specifically to the target DNA sequences adjacent to each other. (B) Ligation and PCR: LPO and RPO joined by ligation enzyme. The linked MLPA probes are amplified by an asymmetric PCR reaction by one universal primer set. The PCR products consist of ds-DNA and ss-DNA.(C) MC analysis: Melting curve assay of the hybridized specific fluorescent detection probe and the ss-DNA of PCR products.</p

Melting curve analysis of one negative control isolate.

<p>(A) Melting peak of the internal control <i>lytA</i> (ROX dye). (B) Melting peak of the internal control <i>cpsA</i> (Cy5 dye).</p

Melting curve analysis of 19F control isolate.

<p>(A) 19F serotype specific melting peak and internal control <i>lytA</i> peak (ROX dye).(B) Melting peak of the internal control <i>cpsA</i> (Cy5 dye).</p

Melting curve analysis of all 12 different fluorescent detection probes.

<p>(A) Melting peaks of five serotype/serogroup-specific fluorescent detection probes 14, 19F, 6, 23F, 4 and internal control <i>lytA</i> probe (ROX dye). (B) Melting peaks of the other five serotype/serogroup-specific fluorescent detection probes 9V, 18, 19A, 15F/15A, 15B/15C, and internal control <i>cpsA</i> probe (Cy5 dye).</p

Additional file 1: of Long non-coding RNA UBE2CP3 enhances HCC cell secretion of VEGFA and promotes angiogenesis by activating ERK1/2/HIF-1Îą/VEGFA signalling in hepatocellular carcinoma

Table S1. Sequences of primers and probe sequences used in this study. (DOC 14 kb

Additional file 2: of Long non-coding RNA UBE2CP3 enhances HCC cell secretion of VEGFA and promotes angiogenesis by activating ERK1/2/HIF-1Îą/VEGFA signalling in hepatocellular carcinoma

Figure S1. (A) The infection efficiencies of lncRNA UBE2CP3 in HepG2 and SMMC-7721 were screened by qRT-PCR. (B) The schematic diagram for the co-culture system. (C) 12 kinds of common angiogenic factors were detected by qRT-PCR.. (TIF 24 kb

Additional file 8: of Metabolic reprogramming-based characterization of circulating tumor cells in prostate cancer

Figure S3. The pairwise comparison matrix used in the AHP model. The weighting coefficients of the criteria layer were calculated on the basis of the maximum eigenvalue using the sum-product method. (TIF 481 kb

Additional file 7: of Metabolic reprogramming-based characterization of circulating tumor cells in prostate cancer

Figure S2. Functions of the differentially expressed genes in glucose and glycogen metabolism. These genes included HK2, PDP2, G6PD, PGK1, PHKA1, PYGL, PDK1, and PKM2. (TIF 1108 kb