Development of an optogenetic gene sensitive to daylight and its implications in vision restoration

Optogenetic gene-mediated therapy for restoring vision is thought to be a useful treatment for blind patients. However, light sensitivity achieved using this gene therapy is inferior to that of daylight vision. To increase light sensitivity, we designed three mutants using a bioinformatics approach. Nucleotide sequences encoding two sites in the extracellular loops (ex1, ex3) of mVChR1 close to simulated ion-conducting pathways were replaced by homologous amino acid-encoding sequences of ChR1 or ChR2. The light sensitivity of ex3mV1 was higher than that of mVChR1 at 405–617 nm. Visual responses were restored in Royal College of Surgeons rats with genetically degenerating photoreceptor cells transfected with ex3mV1Co, wherein transmembrane of sixth (TM6) in ex3mV1 was additionally replaced with the corresponding domain of CoChR; these rats responded to light in the order of μW/mm2. Thus, ex3mV1Co might be useful for the restoration of advanced visual function.

Patagonin-CRISP: Antimicrobial Activity and Source of Antimicrobial Molecules in Duvernoy’s Gland Secretion (Philodryas patagoniensis Snake)

Snake venom contains a variety of toxins with a range of biological activity, among these toxins cysteine-rich secreted proteins (CRISPs) can be found. The proteins of this family have masses of 20–30 kDa and display homologous amino acid sequences containing 16 cysteine residues, forming eight disulfide bonds. Some of these proteins have been explored, characterized, and described in terms of their activity; however, little is known about their range of activities. A search for new antimicrobial molecules is ongoing, as the number of microbial strains resistant to available antibiotics is increasing. We identified antimicrobial activity in the secretion of Duvernoy's gland of the rear-fanged Philodryas patagoniensis. Fractions of this venom were subjected to reverse-phase high performance liquid chromatography and analyzed to determine their antimicrobial activity with a liquid broth inhibition assay. One of the fractions presented activity against a Gram-negative bacterium and a filamentous fungus. This fraction was analyzed with LC-MS/MS, and a protein of 24,848.8 Da was identified. Database searches allowed us to identify it as a CRISP due to the presence of some unique fragments in the molecule. We called it patagonin-CRISP, as the same protein in the venom of P. patagoniensis had previously been characterized as having a different biological activity. Patagonin-CRISP presented activity at very low concentrations and showed no cytotoxic activity. This is the first time that antimicrobial activity has been identified for P. patagoniensis venom or for a CRISP family protein.

COVID-19 Infection and Previous BCG Vaccination Coverage in the Ecuadorian Population

The Bacillus Calmette–Guérin (BCG) is a well-known vaccine with almost a century of use, with the apparent capability to improve cytokine production and epigenetics changes that could develop a better response to pathogens. It has been postulated that BCG protection against SARS-CoV-2 has a potential role in the pandemic, through the presence of homologous amino acid sequences. To identify a possible link between BCG vaccination coverage and COVID-19 cases, we used official epidemic data and Ecuadorian Ministry of Health and Pan American Health Organization vaccination information. BCG information before 1979 was available only at a national level. Therefore, projections based on the last 20 years were performed, to compare by specific geographic units. We used a Mann–Kendall test to identify BCG coverage variations, and mapping was conducted with a free geographic information system (QGIS). Nine provinces where BCG vaccine coverage was lower than 74.25% show a significant statistical association (χ2 Pearson’s = 4.800, df = 1, p = 0.028), with a higher prevalence of cases for people aged 50 to 64 years than in younger people aged 20 to 49 years. Despite the availability of BCG vaccination data and the mathematical models needed to compare these data with COVID-19 cases, our results show that, in geographic areas where BCG coverage was low, 50% presented a high prevalence of COVID-19 cases that were young; thus, low-coverage years were more affected.

NBS-LRR-containing class of salicylic acid-induced gene transcript in rye

NBS-LRR-type disease resistance gene-like cDNA, induced by salicylic acid (SA) was cloned from rye Secalecereale L. (2n = 14RR) var. Petkus, which has rust resistance genes such as Lr26, Sr31 andRr9. We designed primers based on the NBS region and performed PCR using Petkus genomic DNA as a template. Next, we TA-cloned a 532-bp DNA fragment containing five homologous amino acid sequences in the NBS region. The SA-treated rye showed strong expression of a transcript of approximately 3.5 knt in the Northern blots probed with the NBS fragment; however, no transcripts were observed with the untreated rye. We constructed a cDNA library of rye var. Petkus treated with SA, and then screened the cDNA library using the TA-cloned NBS fragments as a probe. The entire nucleotide sequence of a full length of rye NBS-LRR-containing class cDNA 3,446 bp was determined.

The Parathyroid Hormone and Peptides Like It. Literature Review

Wide prevalence of the parathyroid glands pathology and the need for new methods of diagnosis and treatment are forcing researchers all over the world to go more deeply into the pathophysiological mechanisms. A parathyroid hormone (PTH) is main cause of mineral disorders. In addition, humans have a family with similar in structure molecules that contribute to the maintenance of calcium and phosphate homeostasis. The family includes PTH, parathyroid hormone-related protein (PTHrP) and tuberoinfundibular peptide 39 (TIP39, also known as PTH2). The genes encoding these peptides have highly homologous amino acid regions in the N-(amino) terminal receptor-binding sites of each family member, as well as the preserved structure of their organization, which seems to be due to the presence of one parent gene. The variety of classical and “non-classical” effects allows to expand the understanding of these substances and consider them as hormones that go beyond the regulation of phosphorus-calcium metabolism. The review provides information on the structure and biosynthesis of these peptides, as well as a wide range of their effects on the human body.

Binding Sites of miR-1273 Family on the mRNA of Target Genes

This study examined binding sites of 2,578 miRNAs in the mRNAs of 12,175 human genes using the MirTarget program. It found that the miRNAs of miR-1273 family have between 33 and 1,074 mRNA target genes, with a free hybridization energy of 90% or more of its maximum value. The miR-1273 family consists of miR-1273a, miR-1273c, miR-1273d, miR-1273e, miR-1273f, miR-1273g-3p, miR-1273g-5p, miR-1273h-3p, and miR-1273h-5p. Unique miRNAs (miR-1273e, miR-1273f, and miR-1273g-3p) have more than 400 target genes. We established 99 mRNA nucleotide sequences that contain arranged binding sites for the miR-1273 family. High conservation of each miRNA binding site in the mRNA of the target genes was found. The arranged binding sites of the miR-1273 family are located in the 5′UTR, CDS, or 3′UTR of many mRNAs. Five repeating sites containing some of the miR-1273 family’s binding sites were found in the 3′UTR of several target genes. The oligonucleotide sequences of miR-1273 binding sites located in CDSs code for homologous amino acid sequences in the proteins of target genes. The biological role of unique miRNAs was also discussed.

Positive Darwinian selection results in resistance to cardioactive toxins in true toads (Anura: Bufonidae)

Members of the Family Bufonidae, true toads, are famous for their endogenously synthesized cardioactive steroids that serve as defensive toxins. Evolution of resistance to these toxins is not understood. We sequenced a key region of the toxin's binding site in the Na + /K + ATPase for relevant taxa representing Hyloidea (including bufonids), Ranoidea and Archaeobatrachia and tested for positive selection in a phylogenetic context. Bufonidae were distinct from other Hyloidea at 4–6 of 12 sites and, with one exception, had a homologous amino acid sequence. Melanophryniscus stelzneri had a distinct sequence, consistent with other independent evidence for a differentiated toxin. Tests within Bufonidae detected positive selection within the binding region, providing, to our knowledge, the first evidence of this type for positive selection within Amphibia. There was no evidence for positive selection on Bufonidae or M. stelzneri lineages. Sequence change in Leptodactylus ocellatus , a leptodactylid predator of Bufonidae, provides a molecular basis for predator resistance possibly associated with gene duplication.