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Efficient Surveillance For Healthcare Associated Infections Spreading Between Hospitals
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Received: 17 March 2021 / Updated: 29 April 2021 / Accepted: 3 May 2021 / Published: 12 May 2021
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Several functions of the Huntington’s disease protein were investigated in wild-type Hemicentrotus pulcherrimus (Hp-Htt) using Hp-Htt antibodies (Ab) against synthetic oligopeptides. Based on immunoblotting, Hp-Htt was detected as a single band in the 350 kDa region from the swimming blast stage to the larval stage. From stage 2-arm plateus (2aPL), however, a small additional band appeared in the region of 165 kDa. Immunohistochemically, Hp-Htt was detected in the center and near the cytoplasm of ectodermal cells in the floating blastula stage, and blastocular cells from the middle gastrula stage. The closed circuit warning (CBAS) has been replaced. There, many CBAS signaling proteins are present in the cytoplasm, such as glutamate decarboxylase. The application of Hp-Htt morpholino (Hp-Htt-MO) caused the reduction of larval arms, which caused a decrease in the synthesis of 5-bromo-2-deoxyuridin (BrdU) in the ectodermal cells of the larvae. Hp-Htt-MO also caused a decrease in ciliary beat function, with an abnormal body structure. These Hp-Htt-MO defects occurred after the establishment of the CBAS system in the larval armor. Therefore, Hp-Htt is involved in the cell proliferation and signaling process that controls the ciliary beat pattern in platypus larvae.
Huntington’s disease (HD) is a neurological disorder in humans that involves involuntary movements and late-onset dementia, caused by mutations in the Huntingtin (HTT) gene. In mutant Htt, the N-terminal region containing the cytosine-adenine-guanine (CAG) repeat is greatly expanded and has been the subject of many neuropsychological studies, including molecular methods [1]. On the other hand, wild-type Htt seems to be necessary for embryogenesis [2], such as ciliogenesis and neurogenesis in Xenopus [3]. It is also involved in extracellular matrix formation [4] and axonal transport in mammals and cellular trafficking in the sea snail Alysia [5]. HTT is essential for the development and long-term survival in Drosophila [6], and is involved in the regulation of the cerebrospinal fluid system in humans [7, 8, 9, 10]. HTT is also present in marine invertebrates, such as the jellyfish Sienna capillata [11], sea snails Aplysia californica, Branchiostoma floridae [12], and the ascidian Ciona intestinalis [13], and basal deuterostomes such as the sea urchin. Heliocidaris erythrogramma (He-Htt) [12, 14], and Strongylocentrotus purpuratus (Sp-Htt) [11, 15, 16]. However, there is no neurological deficit in these animals, and CAG has not been found to be involved in the lesions mentioned above to date. In fact, according to in situ hybridization studies, He-HTT and Sp-HTT are not expressed in nerve tissue, such as the radial nerve in children, but in mesodermal origin, including hydrocoils, radial canals, and ampullae. [15, 16]. Thus, the evolutionarily conserved functions of wild-type HTT, if any, are still unknown.
Recent advances in genetic activity such as Echinobase (https://www.echinobase.org/entry/), (accessed: 3 March 2021), including the sea urchin Strongylocentrotus purpuratus and many other marine invertebrates . Specifically for the sea urchin, Hemisentrotus pulcherrimus Genome and Transcriptome Database (HpBase: http://cell-innovation.nig.ac.jp/Hpul/) (accessed 3 March 2021) which revealed H. pulcherrimus HTT, Hp-Htt, activation. let’s continue to study the evolution of Htt activities.
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Based on these sea urchin DNA data and subsequent experiments, the present study aims to determine the role of the Hp-Htt protein in the early stages of development, from the swimblastula stage (sBL) to the 6-arm pluteus stage ( 6aPL). We started by introducing a dynamic expression model of Hp-Htt (1). These include (2) analysis of the immunohistochemical expression patterns of the protein, (3) analysis of Hp-Htt activity in embryos using Hp-Htt morpholino (Hp-Htt-MO). negative morphogenic focusing on cell growth activities, and finally (5) the study of larval swimming behavior based on the study of ciliary beat activity and Hp-Htt-MO.
Anti-Hp-Htt antibody (Ab) detected one band from the sBL level to the larval level (pL) around the 350 kDa region (Figure 1, arrow A). However, from the 2-arm pluteus stage (2aPL) to the 4aPL stage, the immunoreactivity of the first large part decreased, with the appearance of a new band of increased immunoreactivity in the region of 165 kDa (Fig. 1, arrow b ). ). . 4aPL, the central part of Hp-Htt, which contains the antigen peptide sequence, is further divided into two new small fragments of 75 kDa and 192 kDa (Figure 1, small arrows). This level is weaker than the first two bands at 350 kDa and 165 kDa. This may suggest continued Hp-Htt dissociation during development, as will be discussed later.
To investigate the possible dissociation of Hp-Htt in and after 2aPL, a protein Hp-Tjp1 (protein junction 1) containing 1402 amino acids and 154.4 kDa (HpBase: http://cell-composition).nig.ac.jp /cgi-bin/Hpul_public/Hpul_annot_search_output.cgi) (accessed: 3 March 2021) and fetal ectodermal tissue [17, 18, 19] were analyzed as loading and degradation control in the lysate. .
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In the pL phase, Tjp1 was detected in the region around 150 kDa (Figure 1, lane 6, arrow C). In step 4aPL, the protein was found in a single region of 150 kDa (Figure 1, line 7). The pretreatment, however, did not show a positive signal (Figure 1, line 8), indicating that the particles derived from Hp-Htt were not detected by Tjp1.
In the ectodermal cells of sBL, positive signals of Hp-Htt were detected in the cytoplasm and nucleus (Figure 2A, B). In the mesenchyme blastulae (mBL), the positive signal of cytoplasmic Hp-Htt spread immediately at the bottom of the cell, but no positive signal was found in the primary mesenchyme cells (Fig. 2C, D*). A positive nuclear Hp-Htt signal was observed in the ectodermal cells in the m (Figure 2D*, E*). At the end of the pL stage, the positive Hp-Htt signal was mainly seen in the blastocular cells (Figure 2F), and many of these cells near the mouth opening expressed glutamic acid decarboxylase (GAD) as well (Figure 2G). Quantitative analysis of the distribution of Hp-Htt-positive cells during early development is consistent with the above-mentioned immunohistochemical findings (Figure 2H). In the MBL, the Hp-Htt-positive signal is confined to the larval ectoderm. Early gastrulae (eG), although a small number of blastocoelar cells showed a positive Hp-Htt signal (Figure 2H, red arrow in eG box), ectodermal cells still dominated significantly (p-value = 0.0146, n = 199) . ). ). From the middle gastrula stage, however, a significant number of mesenchyme cells showed positive Hp-Htt staining. At the pL stage, blastocoelar Hp-Htt-positive cells increased the number of Hp-Htt-positive signals in the ectoderm (p-value = 0.10061, n = 196).
From the 2aPL stage, serotonin is strongly expressed in the apical ganglion and Hp-Htt has a positive signal in the blastocular cells around the apical ganglion (Figure 3A-C), and in the arms of the larva. 3A, B, F, arrow). In the blastocoel, these Hp-Htt-positive cells are distributed in the basal region of the ectoderm (Figure 3A, B). In the apical ganglion region, serotonin- and Hp-Htt-positive signals were found in the corresponding region without overlap with each other (Figure 3C, D*). In 4aPL, a positive signal of Hp-Htt was detected as one of the bound proteins in the GAD-expressing chain-binding complex (CBAS) (Figure 3E, G). Although the regions expressing GAD and Hp-Htt were close to the axial region of CBAS, their positive signals were not in the cytoplasm (Figure 3F, G).
Duizenden Mensen Zonder Huisarts Door Huisartsentekort
Hp-Htt is found in the cytoplasm of the perikaryon but not in the nucleus.
