Self-transfecting GMO-PMO and PMO-GMO chimeras enable gene silencing in vitro and in vivo zebrafish model and NANOG Inhibition Induce the Apoptosis in Breast and Prostate Cancer Cells

Phosphorodiamidate Morpholino Oligonucleotides (PMOs)-based antisense reagents cannot enter inside cells by itself without the help of any delivery technique which is the last hurdle for their clinical applications. To overcome this limitation, a self-transfecting GMO-PMO or PMO-GMO chimeras has been explored as a gene silencing reagent where GMO stands for guanidinium morpholino oligonucleotides which linked either at the OH- or NH-end of PMOs. GMO not only facilitates cellular internalization of such chimeras but also participates in Watson-Crick base pairing during gene silencing in ShhL2 cells when designed against mGli1 and compared with scrambled GMO-PMO where mutations were made only to the GMO part. GMO-PMO-mediated knockdown of no tail gene resulted no tail-dependent phenotypes in zebrafish and worked even after the delivery at 16-, 32- and 64-cell stages which were previously unachievable by regular PMO. Furthermore, GMO-PMO chimeras has shown the inhibition of NANOG, a key regulator of self-renewal and pluripotency of both embryonic and cancer stem cells. Its inhibition influences on the expression of other cancer related proteins and the respective phenotypes in breast cancer cells and increases the therapeutic potential of taxol. To the best of our knowledge, this is the first report on the self-transfecting antisense reagents since the discovery of guanidinium linked DNA (DNG) and most effective among the all cell-penetrating PMOs reported till date expected to solve the longstanding problem of PMO delivery. In principle, this technology could be useful for the inhibition of any target gene without using any delivery vehicle and should have applications in the fields of antisense therapy, diagnostic and nanotechnology area.

Tail or no Tail? Applicability of the Long Tail Theory to the German Online Media Market

The Long Tail theory serves as an explanation for market change triggered through evolving online market conditions. However, it is often referred to without empirical validation, or with inconsistent findings on its applicability. Therefore, this paper analyses the applicability of the Long Tail theory to German online media offerings as of 2014 and 2016, focusing on offerings that serve information purposes (information offerings). Based on a unique dataset of commercially oriented online media offerings (Longitudinal IntermediaPlus 2014–2016), an analysis for three Long Tail characteristics (variety increase, opposing concentration between hit and niche offerings and niche market share increase) is conducted. By additionally distinguishing between political and entertainment-oriented information offerings as boundaries of democratized online market conditions the analysis reveals that the Long Tail theory is not fully applicable to any market sample, and especially not to politically oriented offerings.

Morphometrics of the indian false vampire bat (Megaderma lyra) from district Jhelum, Pakistan

During the present study thirteen Megaderma lyra bats were observed roosting in dark, domed shaped room of Rohtas Fort, district Jhelum. Out of these, six specimens were captured from the roosting site, using hand net. All captured specimens were male. These bats were identified through their unique facial features, an erect and elongated nose-leaf, large oval ears that joined above the forehead and no tail. Mean head and body length of captured specimens was 80 mm, forearm length was 67 mm while average lengths of 3rd, 4th and 5th metacarpals were 51.73 mm, 55.17 mm and 60.42 mm, respectively. Mean skull length was 29.84 mm, breadth of braincase was 12.77 mm. Average Penis length of two specimens was 6.6 mm and total bacular length was 3.08 mm respectively. This is the first record of Megaderma lyra from district Jhelum.

Sturnira parvidens (Chiroptera: Phyllostomidae)

The little yellow-shouldered Mesoamerican bat, Sturnira parvidens Goldman, 1917, is a medium-sized yellow-shouldered bat with no tail, a vestigial uropatagium, and reddish or yellowish patches on the shoulders. It lives in tropical habitats associated with lower and mid-elevations from northern Costa Rica to Mexico, and it is one of 24 described species in the genus Sturnira. Although S. parvidens is more common in disturbed areas because it mainly feeds on pioneer plants, it uses day roosts located in mature forest or in areas with advanced successional stages. It is an abundant species and is listed as “Least Concern” by the International Union for Conservation of Nature and Natural Resources.

Zero Waste Process for Direct Forming Process of Nail Spikes

This study aims to plan the no-tail, i.e. zero waste, automatic cold forging model through the innovative development of a die to directly produce finished nail tails without second processing. It could enhance production efficiency, reduce production costs, and conform to the new model of zero waste under “circular economy”.

Synthesis of Phosphorodiamidate Morpholino Oligonucleotides Using Trityl and Fmoc Chemistry-A New Method Amenable to Automated Synthesizer

<p>Phosphorodiamidatemorpholino oligonucleotides (PMO) are routinely used for gene silencing and the recently developed PMO-based drug “Exondys51” has highlighted the importance of PMO as excellent antisense reagents. However, the synthesis of PMO has remained challenging. Here a method for the synthesis of PMO using either trityl or Fmoc-protected active morpholino monomers using chlorophosphoramidate chemistry in the presence of a suitable coupling agent on a solid support has been reported. After screening several coupling agents (tetrazole, 1,2,4-triazole, ETT, iodine, LiBr and dicyanoimidazole), ETT and iodine were found to be suitable for efficient coupling. Fmoc chemistry was not known for PMO synthesis because the preparation of Fmoc-protected chlorophosphoramidate monomers was not trivial. Synthesis of Fmoc-protected activated monomers and their use in PMO synthesis is reported for the first time. 25-mer PMO has been synthesized using both the methods and validated <i>in vivo</i> in the zebrafish model by targeting the <i>no tail</i> gene. Methods have been transferred in DNA synthesizer which has become user friendly for PMO synthesis and opened a new avenue to explore PMO for various applications. Fmoc chemistry could be suitable for scalable approach of PMO synthesis using peptide synthesizer as it is a neutral oligomer like peptide.</p>

Synthesis of Phosphorodiamidate Morpholino Oligonucleotides Using Trityl and Fmoc Chemistry-A New Method Amenable to Automated Synthesizer

<p>Phosphorodiamidatemorpholino oligonucleotides (PMO) are routinely used for gene silencing and the recently developed PMO-based drug “Exondys51” has highlighted the importance of PMO as excellent antisense reagents. However, the synthesis of PMO has remained challenging. Here a method for the synthesis of PMO using either trityl or Fmoc-protected active morpholino monomers using chlorophosphoramidate chemistry in the presence of a suitable coupling agent on a solid support has been reported. After screening several coupling agents (tetrazole, 1,2,4-triazole, ETT, iodine, LiBr and dicyanoimidazole), ETT and iodine were found to be suitable for efficient coupling. Fmoc chemistry was not known for PMO synthesis because the preparation of Fmoc-protected chlorophosphoramidate monomers was not trivial. Synthesis of Fmoc-protected activated monomers and their use in PMO synthesis is reported for the first time. 25-mer PMO has been synthesized using both the methods and validated <i>in vivo</i> in the zebrafish model by targeting the <i>no tail</i> gene. Methods have been transferred in DNA synthesizer which has become user friendly for PMO synthesis and opened a new avenue to explore PMO for various applications. Fmoc chemistry could be suitable for scalable approach of PMO synthesis using peptide synthesizer as it is a neutral oligomer like peptide.</p>

Effect of tail fin loss on swimming capability and tail beat frequency of juvenile black carp Mylopharyngodon piceus

Fin clipping is a common practice in fisheries management, and hatchery fish are often marked this way. In the wild, the tail (caudal) fin may be damaged or lost to predation or disease. Because the tail fin is important to fish swimming behavior and ability, this study was designed to examine the effects of partial and complete loss of the tail fin on the swimming ability of juvenile black carp Mylopharyngodon piceus. Swimming speed and tail beat frequency were measured for 3 groups (intact tail fin, partial tail fin, no tail fin) using a stepped velocity test conducted in a fish respirometer. We found that critical swimming speed (Ucrit) and burst speed (Uburst) decreased slightly in the partial fin group and significantly in the no fin group. In the group with no tail fin, Uburst decreased more than Ucrit, clearly reducing the ability to avoid predators. Moreover, mean tail beat frequency (TBFmean), Ucrit and Uburst all decreased slightly in the partial fin group and significantly in the no fin group. A decrease in tail beat force and TBF both reduce swimming capability. These findings contribute to developing our understanding of the relationship between fish tail fins and swimming.

Self-force and the Huygens principle

We consider a relation between the Huygens Principle (HP) in gravity and the self-interaction force. We show that the self-force for an electric particle in the plane gravitational wave space-time has no tail term even the vector Green function does not obey the HP. The reason for this observation is that even vector potential does not obey the HP, the electromagnetic field does obey.

Greb1 is required for axial elongation and segmentation in vertebrate embryos

ABSTRACTDuring vertebrate embryonic development, the formation of axial structures is driven by a population of stem-like cells that reside in a region of the tailbud called the chordoneural hinge (CNH). We have compared the CNH transcriptome with those of surrounding tissues and shown that the CNH and tailbud mesoderm are transcriptionally similar, and distinct from the presomitic mesoderm. Amongst CNH-enriched genes are several that are required for axial elongation, including Wnt3a, Cdx2, Brachyury/T and Fgf8, and androgen/estrogen receptor nuclear signalling components such as Greb1. We show that the pattern and duration of tailbud Greb1 expression is conserved in mouse, zebrafish, and chicken embryos, and that Greb1 is required for axial elongation and somitogenesis in zebrafish embryos. The axial truncation phenotype of Greb1 morphant embryos is explained by much reduced expression of No tail (Ntl/Brachyury) which is required for axial progenitor maintenance. Posterior segmentation defects in the morphants (including misexpression of genes such as mespb, myoD and papC) appear to result, in part, from lost expression of the segmentation clock gene, her7.