Ley 2108 de 2021: Una mirada al internet como servicio público desde la esencialidad y su universalidad.

Today, the Internet is a public service with easy and fast access that brings with it multiple benefits for its users, such as instantaneous communication from anywhere in the world and instant accessibility to a multitude of information, this generates that, the great part of the world population wants to have access to it, forcing governments to create regulations that regulate this public service for an efficient provision of the service. For this reason, in this legal-descriptive investigation, the implications of the issuance of Law 2108 of 2021, which consecrated the internet as an essential and universal public service in Colombia, will be addressed.En la actualidad, el internet es un servicio público de fácil y rápido acceso que trae consigo múltiples beneficios para sus usuarios, tales como, la comunicación desde cualquier lugar del mundo instantáneamente y la accesibilidad a multitud de información al instante, esto, genera que, la gran parte de la población mundial quiera tener acceso a este, obligando a los gobiernos a la creación de normatividad que regule este servicio público para una eficaz prestación del servicio. Por ello, en la presente investigación jurídica- descriptiva se abordarán las implicaciones que trajo consigo la expedición de la Ley 2108 del 2021 que consagró el internet como servicio público esencial y universal en Colombia

Toward the Discovery of Biological Functions Associated with the Mechanosensor Mtl1p of \u3ci\u3eSaccharomyces cerevisiae\u3c/i\u3e via Integrative Multi-OMICs Analysis

Functional analysis of the Mtl1 protein in Saccharomyces cerevisiae has revealed that this transmembrane sensor endows yeast cells with resistance to oxidative stress through a signaling mechanism called the cell wall integrity pathway (CWI). We observed upregulation of multiple heat shock proteins (HSPs), proteins associated with the formation of stress granules, and the phosphatase subunit of trehalose 6-phosphate synthase which suggests that mtl1Δ strains undergo intrinsic activation of a non-lethal heat stress response. Furthermore, quantitative global proteomic analysis conducted on TMT-labeled proteins combined with metabolome analysis revealed that mtl1Δ strains exhibit decreased levels of metabolites of carboxylic acid metabolism, decreased expression of anabolic enzymes and increased expression of catabolic enzymes involved in the metabolism of amino acids, with enhanced expression of mitochondrial respirasome proteins. These observations support the idea that Mtl1 protein controls the suppression of a non-lethal heat stress response under normal conditions while it plays an important role in metabolic regulatory mechanisms linked to TORC1 signaling that are required to maintain cellular homeostasis and optimal mitochondrial function

Characterization of Histone Genes from the Bivalve Lucina Pectinata

Lucina pectinata is a clam that lives in sulfide-rich environments and houses intracellular sulfide-oxidizing endosymbionts. To identify new Lucina pectinata proteins, we produced libraries for genome and transcriptome sequencing and assembled them de novo. We searched for histone-like sequences using the Lucina pectinata histone H3 partial nucleotide sequence against our previously described genome assembly to obtain the complete coding region and identify H3 coding sequences from mollusk sequences in Genbank. Solen marginatus histone nucleotide sequences were used as query sequences using the genome and transcriptome assemblies to identify the Lucina pectinata H1, H2A, H2B and H4 genes and mRNAs and obtained the complete coding regions of the five histone genes by RT-PCR combined with automated Sanger DNA sequencing. The amino acid sequence conservation between the Lucina pectinata and Solen marginatus histones was: 77%, 93%, 83%, 96% and 97% for H1, H2A, H2B, H3 and H4, respectively. As expected, the H3 and H4 proteins were the most conserved and the H1 proteins were most similar to H1′s from aquatic organisms like Crassostrea gigas, Aplysia californica, Mytilus trossulus and Biomphalaria glabrata. The Lucina pectinata draft genome and transcriptome assemblies, obtained by semiconductor sequencing, were adequate for identification of conserved proteins as evidenced by our results for the histone genes

Characterization of Histone Genes from the Bivalve Lucina Pectinata

Lucina pectinata is a clam that lives in sulfide-rich environments and houses intracellular sulfide-oxidizing endosymbionts. To identify new Lucina pectinata proteins, we produced libraries for genome and transcriptome sequencing and assembled them de novo. We searched for histone-like sequences using the Lucina pectinata histone H3 partial nucleotide sequence against our previously described genome assembly to obtain the complete coding region and identify H3 coding sequences from mollusk sequences in Genbank. Solen marginatus histone nucleotide sequences were used as query sequences using the genome and transcriptome assemblies to identify the Lucina pectinata H1, H2A, H2B and H4 genes and mRNAs and obtained the complete coding regions of the five histone genes by RT-PCR combined with automated Sanger DNA sequencing. The amino acid sequence conservation between the Lucina pectinata and Solen marginatus histones was: 77%, 93%, 83%, 96% and 97% for H1, H2A, H2B, H3 and H4, respectively. As expected, the H3 and H4 proteins were the most conserved and the H1 proteins were most similar to H1′s from aquatic organisms like Crassostrea gigas, Aplysia californica, Mytilus trossulus and Biomphalaria glabrata. The Lucina pectinata draft genome and transcriptome assemblies, obtained by semiconductor sequencing, were adequate for identification of conserved proteins as evidenced by our results for the histone genes

Characterization and Expression of the <i>Lucina pectinata</i> Oxygen and Sulfide Binding Hemoglobin Genes

<div><p>The clam <i>Lucina pectinata</i> lives in sulfide-rich muds and houses intracellular symbiotic bacteria that need to be supplied with hydrogen sulfide and oxygen. This clam possesses three hemoglobins: hemoglobin I (HbI), a sulfide-reactive protein, and hemoglobin II (HbII) and III (HbIII), which are oxygen-reactive. We characterized the complete gene sequence and promoter regions for the oxygen reactive hemoglobins and the partial structure and promoters of the HbI gene from <i>Lucina pectinata</i>. We show that HbI has two mRNA variants, where the 5’end had either a sequence of 96 bp (long variant) or 37 bp (short variant). The gene structure of the oxygen reactive Hbs is defined by having 4-exons/3-introns with conservation of intron location at B12.2 and G7.0 and the presence of pre-coding introns, while the partial gene structure of HbI has the same intron conservation but appears to have a 5-exon/ 4-intron structure. A search for putative transcription factor binding sites (TFBSs) was done with the promoters for HbII, HbIII, HbI short and HbI long. The HbII, HbIII and HbI long promoters showed similar predicted TFBSs. We also characterized MITE-like elements in the HbI and HbII gene promoters and intronic regions that are similar to sequences found in other mollusk genomes. The gene expression levels of the clam Hbs, from sulfide-rich and sulfide-poor environments showed a significant decrease of expression in the symbiont-containing tissue for those clams in a sulfide-poor environment, suggesting that the sulfide concentration may be involved in the regulation of these proteins. Gene expression evaluation of the two HbI mRNA variants indicated that the longer variant is expressed at higher levels than the shorter variant in both environments.</p></div

Alignment of HbI cDNA sequences obtained by RT-PCR.

<p>HbI cDNA was amplified with forwards primers designed from short and long variant and a common reverse primer, the sequences are named short and long, respectively. Samples from 1 to 3 correspond to RNA isolated from clams kept in a fish tank and samples 4 to 6 correspond to RNA isolated from clams harvested from their natural environment. F indicates that the sequence was obtained with the corresponding forward primer used for the cDNA amplification and R indicates that the sequence was obtained with the reverse primer. The first three bases coding for the first amino acid at the protein level are marked as start and the last marked as end, both highlighted in blue. Each position that presented different nucleotides are indicated by the number of this position in the reported mRNA for each Hb. The alignments were generated using Clustal W [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0147977#pone.0147977.ref023" target="_blank">23</a>] and visualized and formatted with GeneDoc [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0147977#pone.0147977.ref035" target="_blank">35</a>].</p

Dot plot comparison of the incomplete HbI gene to the HbII gene.

<p>A repetitive region in HbI intron and HbII promoter sequence and introns was detected. The HbII gene structure is represented in the horizontal axis and the HbI incomplete gene structure is shown in the vertical axis.</p

Relative quantitation of mRNAs of <i>Lucina pectinata</i>’s Hbs in all tissues examined for both conditions.

<p>Data is represented as aligned dot plot using logarithmic scale, showing mean (horizontal bars) and SEM (whiskers). One group was harvested in their sulfide-rich mud natural environment (labeled on X axis as Natural environment) and another group of clams were kept in a fish tank in fresh sea water (labeled on X axis as Fish Tank). Significant differences are indicated with an asterisk at p < 0.05, and with two asterisks at p < 0.01. Gene expression analysis at the mRNA level of <i>L</i>. <i>pectinata</i> hemoglobins (Hbs) was measured using Real Time RT-PCR. <b>A)</b> Hbs expression in ctenidia tissue; <b>B)</b> Hbs expression in mantle tissue; <b>C)</b> Hbs expression in muscle tissue; <b>D)</b> Hbs expression in foot tissue; <b>E)</b> Hbs expression in visceral mass tissue.</p

<i>Lucina pectinata</i> hemoglobin gene structures.

<p>Schematic diagram of <i>L</i>. <i>pectinata</i>’s hemoglobin genes. The exon regions are represented by boxes and introns as dashed lines. The orange triangles represent the location of MITE-like element found in these genes. The sequences preceding exon(s) 1 are the promoter regions obtained by the GW method. <b>A)</b> Schematic diagram of HbII gene structure. Shown are the localization and length of the XL-PCR products amplified to determine the intron sizes and overlapping PCR products used to verify the 3’ end of the gene. The total length of the four exons and three introns in the <i>L</i>. <i>pectinata</i> HbII gene is 8,055 bp. <b>B)</b> Schematic diagram of HbIII gene structure. The exon regions, the localization and length of the XL-PCR products and overlapping PCR products are indicated as for the HbII gene in A. The total length of the four exons and three introns in the <i>L</i>. <i>pectinata</i> HbIII gene is 4,277 bp. <b>C)</b> Proposed schematic diagram of HbI partial gene structure. The localization and length of PCR amplified to determine the intron sizes and overlapping products are shown. HbI incomplete gene length (from incomplete intron 1_a to the end of exon 4) is 4,341 bp. Alternate first exons are label exon 1_L and exon 1_S.</p

Relative quantitation of the two HbI mRNAs variants in ctenidia tissue for both studied conditions.

<p>Data is represented as aligned dot plot using logarithmic scale, showing mean (horizontal bars) and SEM (whiskers). One group was harvested in their sulfide rich mud natural environment (labeled on X axis as Natural environment) and another group were kept in a fish tank in fresh sea water (labeled on X axis as Fish Tank). Significant differences are indicated with an asterisk at P < 0.05, and with two asterisks at P < 0.01. Gene expression analysis at the mRNA level of L. pectinata hemoglobins (Hbs) was measured using Real Time RT-PCR.</p