Phosphate-Methylated Oligonucleotides as a Novel Primer for PCR and RT-PCR.

This chapter introduces neutralized DNA (nDNA) as a novel design for the primers of PCR and RT-PCR by methylating phosphate groups of some oligonucleotides in their structures. It starts with an introduction of the nDNA which possesses an electrically chimeric neutral backbone as well as the proposed standards in designing nDNA as a novel primer for PCR and RT-PCR , concluded from various experimental results presented afterward. The primary content comprises empirical data from PCR to compare nDNA and unmodified DNA as primers in terms of ability to distinguish and amplify mismatch templates, activities of polymerase enzymes, melting temperature of double-stranded sequences, and the trials and discussions on various modified positions of the nDNA primers. In summary, nDNA exhibited outstanding performance as a primer for PCR and RT-PCR , compared to unmodified DNA, and is expected to be expanded in diverse applications which require enhanced specificity.

The experiences of nurses and midwives who provide surgical abortion care: A qualitative systematic review.

AIM: To synthesize qualitative evidence on nurses' and midwives' experiences in the provision of surgical abortion care. We address three specific questions: (a) what are the experiences of nurses and midwives in surgical abortion care? (b) what are their responses and coping strategies? (c) what are the deficiencies in surgical abortion care? DESIGN: Qualitative studies were synthesized using Thomas and Harden's qualitative thematic synthesis method. DATA SOURCES: Electronic databases, including PubMed, Embase, CINAHL, PsycINFO, Scopus and Web of Science were searched. Grey literature using ProQuest was searched. The databases were searched from inception to 5 August 2020. REVIEW METHODS: The SPIDER (Sample, Phenomenon of Interest, Design, Evaluation and Research type) search tool was used in the literature search. Data synthesis was conducted using the three-stage thematic synthesis method described by Thomas and Harden. RESULTS: 966 studies were identified in the initial search and 18 studies were included. Four analytical themes were generated: 'Providing abortion care requires high emotional labour'; 'Professionalism of abortion care providers'; 'Initiatives in professional development' and 'Improving directions for high-quality abortion care'. CONCLUSION: Nurses and midwives indicated that they require support to enhance psychological health and improve professional skills. Hospital managers should organize regular debriefing or structured group workshops for exchange of practical experiences and strengthening emotional support. More research is required to establish comprehensive training related to abortion care for nurses and midwives. The findings demonstrate that optimization of abortion services should start from hospital management models, pain management and bereavement care. IMPACT: Understanding the experiences of nurses and midwives in abortion provision will inform future clinical practice in surgical abortion care, which would be helpful in improving the professionalism and confidence of abortion providers. Our findings have implications for the training, development of policies and standards for surgical abortion care for nurses and midwives.

Designed phosphate-methylated oligonucleotides as PCR primers for SNP discrimination.

Polymerase chain reaction (PCR) is a powerful technique for the detection and quantification of nucleic acids and has enormous applications to research in molecular biology. Certain inherited diseases, caused by single nucleotide mutations, however, are difficult to identify by PCR, using DNA primers and probes, in a situation where a false diagnosis may lead to incorrect or delayed treatment. With the aim of enhancing the specificity of PCR, we used novel chemically synthesized oligonucleotides containing site-specific methyl phosphotriester (MPTE) inter-nucleoside linkage(s) as primers and probes. The methyl phosphotriester linkages carry no charge, so the reduction in the electrostatic repulsion of an MPTE-DNA/DNA duplex shows stronger hybridization affinity compared to a DNA/DNA duplex. However, the electrosteric effects introduced by the methyl group may result in instability of the double-stranded DNA (dsDNA) formed. With the use of specific MPTE modification sites and optimization of the number of MPTE modifications, greater delta melting temperature (ΔTm) may be obtained, in concert with adjustment of PCR operating conditions, especially with respect to the annealing temperature, to achieve more discriminatory results between the target template and the perfectly matched primer and the mismatched primer. In single nucleotide polymorphism (SNP) genotyping, the results demonstrated that MPTE-modified probes can improve specificity. In summary, MPTE-modified oligonucleotides are a promising DNA analog applied to PCR primers and probes to enhance the specificity and to provide more precise detection results for various applications, such as for genetic diagnosis. In summary, two common DNA polymerases we tested could successfully recognize the MPTE-modified primers and probes. Under the optimal operating conditions, MPTE modification has the ability to improve the discrimination of single nucleotide polymorphism by increasing the ΔTm of the perfect match and mismatch sequences and to provide more precise detection results for various applications, such as genetic diagnosis.

Cu-Catalyzed [3 + 3] Annulation for the Synthesis of Pyrimidines via ß-C(sp3)-H Functionalization of Saturated Ketones.

A novel, efficient, and facile approach for the synthesis of structurally important pyrimidines has been successfully developed by Cu-catalyzed and 4-HO-TEMPO-mediated [3 + 3] annulation of commercially available amidines with saturated ketones. This method provides a new protocol for the synthesis of pyrimidines by a cascade reaction of oxidative dehydrogenation/annulation/oxidative aromatization via direct ß-C(sp3)-H functionalization of saturated ketones followed by annulation with amidines.