A Mixed Finite Element Formulation for the Conservative Fractional Diffusion Equations

We consider a boundary-value problem of one-side conservative elliptic equation involving Riemann-Liouville fractional integral. The appearance of the singular term in the solution leads to lower regularity of the solution of the equation, so to the lower order convergence rate for the numerical solution. In this paper, by the dividing of equation, we drop the lower regularity term in the solution successfully and get a new fractional elliptic equation which has full regularity. We present a theoretical framework of mixed finite element approximation to the new fractional elliptic equation and derive the error estimates for unknown function, its derivative, and fractional-order flux. Some numerical results are illustrated to confirm the optimal error estimates

Development of novel chemically modified antisense oligonucleotides for tackling type 2 diabetes and Duchenne muscular dystrophy

Therapeutic nucleic acids such as antisense oligonucleotides (AOs) can specifically bind to target RNA and manipulate gene expression through different mechanism of actions, including RNase H mediated RNA degradation, splice modulation, and translational repression. So far, the United States Food and Drug Administration has approved seven AO drugs for the therapies of different human diseases. Among them, four drugs (eteplirsen, golodirsen, nusinersen, milasen) modulate splicing, and rescue the production of functional essential proteins by restoring ruined open reading frames due to mutations within genes. This thesis explores the potential of novel chemically-modified splice-modulating AOs in tackling type 2 diabetes (T2D) and Duchenne muscular dystrophy (DMD). Chapter one provides an introduction of different types of therapeutic nucleic acids. Chapter two focused on developing AOs targeting protein tyrosine phosphatase 1B (PTP1B), a validated therapeutic target of T2D. An AO candidate, PTPN1 1E2A (+5+29), was identified to efficiently skip exon-2 of PTPN1 RNA inducing premature termination codons in exon-3, thereby reducing the production of full-length, functional PTP1B proteins. Chapter three presents the work on design, synthesis and evaluation of splice-modulating chimeric AOs containing novel nucleotide analogues for exon skipping in H2K mdx myoblast, using DMD as a disease model. Morpholino nucleic acid modified 2'-O-Methyl (2'-OMe) AO mixmer exhibited comparable exon skipping ability as uniformly modified 2'-OMe AO. Locked nucleic acid (LNA) modified 2'-Fluoro (2'-F) AOs showed improved drug-like properties than 2'-OMe modified 2'-F AOs. In summary, the thesis expands the applicability of AOs in the therapy of T2D through downregulating PTP1B protein expression by splice modulation; the thesis also expands the scope of utilizing nucleotide analogues in splice modulation application by constructing AO chimeras, demonstrating the feasibility of using these analogues for optimizing AOs in terms of improved drug-like properties

Variation Characteristics of Rainfall in the Pre-Flood Season of South China and Its Correlation with Sea Surface Temperature of Pacific

The characteristics of rainfall variation in the pre-flood season of South China (PFSSC) and its correlation with the sea surface temperature (SST) of the Pacific are studied in this paper. The results show that in the last 50 years, rainfall in PFSSC clearly has interannual and interdecadal oscillations, primarily in the 4a and 8a cycles. Interannual correlation analysis indicate that the rainfall in PFSSC displays a significantly negative correlation with the SST of the warm pool region in January–March and April–June. The interdecadal correlation analysis reveals that the rainfall in PFSSC is negatively correlated to the SST of the warm pool region, but has a significant positive interdecadal correlation with the Middle Eastern Pacific Ocean. For NINO1 + 2 and NINO3 regions, when the background ocean temperature is warm, the SST is significantly positively correlated to the rainfall in PFSSC; however, when the background ocean temperature is cold, there is no significant correlation between the two, even the correlation coefficients are negative. For the warm pool region, the SST demonstrates a significantly negative correlation to the rainfall in PFSSC, which is not dependent on the background SST. It is a remarkable fact that under the different SST backgrounds, the interannual variation of SST will bring different atmospheric response, and it is the reason that under the warm SST background, the correlation is more significant between the SST in tropical Pacific and the rainfall in PFSSC. Under the background of global warming, more attention should be given to study the rainfall in PFSSC and its correlation with the SST in the eastern tropical Pacific

Radiolabeling of Nucleic Acid Aptamers for Highly Sensitive Disease-Specific Molecular Imaging

Aptamers are short single-stranded DNA or RNA oligonucleotide ligand molecules with a unique three-dimensional shape, capable of binding to a defined molecular target with high affinity and specificity. Since their discovery, aptamers have been developed for various applications, including molecular imaging, particularly nuclear imaging that holds the highest potential for the clinical translation of aptamer-based molecular imaging probes. Their easy laboratory production without any batch-to-batch variations, their high stability, their small size with no immunogenicity and toxicity, and their flexibility to incorporate various functionalities without compromising the target binding affinity and specificity make aptamers an attractive class of targeted-imaging agents. Aptamer technology has been utilized in nuclear medicine imaging techniques, such as single photon emission computed tomography (SPECT) and positron emission tomography (PET), as highly sensitive and accurate biomedical imaging modalities towards clinical diagnostic applications. However, for aptamer-targeted PET and SPECT imaging, conjugation of appropriate radionuclides to aptamers is crucial. This review summarizes various strategies to link the radionuclides to chemically modified aptamers to accomplish aptamer-targeted PET and SPECT imaging

Radiolabeling of Nucleic Acid Aptamers for Highly Sensitive Disease-Specific Molecular Imaging

Aptamers are short single-stranded DNA or RNA oligonucleotide ligand molecules with a unique three-dimensional shape, capable of binding to a defined molecular target with high affinity and specificity. Since their discovery, aptamers have been developed for various applications, including molecular imaging, particularly nuclear imaging that holds the highest potential for the clinical translation of aptamer-based molecular imaging probes. Their easy laboratory production without any batch-to-batch variations, their high stability, their small size with no immunogenicity and toxicity, and their flexibility to incorporate various functionalities without compromising the target binding affinity and specificity make aptamers an attractive class of targeted-imaging agents. Aptamer technology has been utilized in nuclear medicine imaging techniques, such as single photon emission computed tomography (SPECT) and positron emission tomography (PET), as highly sensitive and accurate biomedical imaging modalities towards clinical diagnostic applications. However, for aptamer-targeted PET and SPECT imaging, conjugation of appropriate radionuclides to aptamers is crucial. This review summarizes various strategies to link the radionuclides to chemically modified aptamers to accomplish aptamer-targeted PET and SPECT imaging

Bichirs employ similar genetic pathways for limb regeneration as are used in lungfish and salamanders

Bichirs are a sister group to sarcopterygian and tetrapods that can fully regenerate their endochondral-skeleton-fins. Histological and transcriptomic comparison approaches have been used to investigate the morphology and genetic basis of bichir lobe-fin regeneration, with strong down-regulation of muscle-related genes and up-regulation of ECM-related genes and developmental genes being observed. Bichir limb regeneration involves similar cellular processes to those employed by lungfish and salamander, with MARCKS-like protein (MLP) that is known to be a putative regeneration-initiating molecule in salamander, also up-regulated in the early stages of bichir lobe-fin regeneration. These gene expression results suggest that limb regeneration pathways in these amphibians have a common ancestral inheritance, consistent with evolution from endochondral-skeleton-fin structures to endochondral-skeleton-limb structures of vertebrates

Antisense Oligonucleotide-Mediated Splice Switching: Potential Therapeutic Approach for Cancer Mitigation

Splicing is an essential process wherein precursor messenger RNA (pre-mRNA) is reshaped into mature mRNA. In alternative splicing, exons of any pre-mRNA get rearranged to form mRNA variants and subsequently protein isoforms, which are distinct both by structure and function. On the other hand, aberrant splicing is the cause of many disorders, including cancer. In the past few decades, developments in the understanding of the underlying biological basis for cancer progression and therapeutic resistance have identified many oncogenes as well as carcinogenic splice variants of essential genes. These transcripts are involved in various cellular processes, such as apoptosis, cell signaling and proliferation. Strategies to inhibit these carcinogenic isoforms at the mRNA level are promising. Antisense oligonucleotides (AOs) have been developed to inhibit the production of alternatively spliced carcinogenic isoforms through splice modulation or mRNA degradation. AOs can also be used to induce splice switching, where the expression of an oncogenic protein can be inhibited by the induction of a premature stop codon. In general, AOs are modified chemically to increase their stability and binding affinity. One of the major concerns with AOs is efficient delivery. Strategies for the delivery of AOs are constantly being evolved to facilitate the entry of AOs into cells. In this review, the different chemical modifications employed and delivery strategies applied are discussed. In addition to that various AOs in clinical trials and their efficacy are discussed herein with a focus on six distinct studies that use AO-mediated exon skipping as a therapeutic strategy to combat cancer

Systematic evaluation of 2′-Fluoro modified chimeric antisense oligonucleotide-mediated exon skipping in vitro

Abstract Antisense oligonucleotide (AO)-mediated splice modulation has been established as a therapeutic approach for tackling genetic diseases. Recently, Exondys51, a drug that aims to correct splicing defects in the dystrophin gene was approved by the US Food and Drug Administration (FDA) for the treatment of Duchenne muscular dystrophy (DMD). However, Exondys51 has relied on phosphorodiamidate morpholino oligomer (PMO) chemistry which poses challenges in the cost of production and compatibility with conventional oligonucleotide synthesis procedures. One approach to overcome this problem is to construct the AO with alternative nucleic acid chemistries using solid-phase oligonucleotide synthesis via standard phosphoramidite chemistry. 2′-Fluoro (2′-F) is a potent RNA analogue that possesses high RNA binding affinity and resistance to nuclease degradation with good safety profile, and an approved drug Macugen containing 2′-F-modified pyrimidines was approved for the treatment of age-related macular degeneration (AMD). In the present study, we investigated the scope of 2′-F nucleotides to construct mixmer and gapmer exon skipping AOs with either 2′-O-methyl (2′-OMe) or locked nucleic acid (LNA) nucleotides on a phosphorothioate (PS) backbone, and evaluated their efficacy in inducing exon-skipping in mdx mouse myotubes in vitro. Our results showed that all AOs containing 2′-F nucleotides induced efficient exon-23 skipping, with LNA/2′-F chimeras achieving better efficiency than the AOs without LNA modification. In addition, LNA/2′-F chimeric AOs demonstrated higher exonuclease stability and lower cytotoxicity than the 2′-OMe/2′-F chimeras. Overall, our findings certainly expand the scope of constructing 2′-F modified AOs in splice modulation by incorporating 2′-OMe and LNA modifications