Gene and miR Expression in the Yellow Fever Mosquito Aedes aegypti

When animals are exposed to new conditions they need to tune the expression of hundreds of genes in specific tissues and at a specific time requires the interplay of multiple processes. Females are required to locate suitable oviposition sites after acquiring a blood meal. Mosquitoes possess a variety of genes responsible for chemosensation, with four families standing out: Olfaction Receptors (ORs), Ionotropic Receptors (IRs), Odorant Binding Proteins (OBPs), and Gustatory Receptors (GRs). To investigate the genes that may be involved in host seeking behavior, mRNA was sequenced from samples of A. aegypti male and female tissues using RNA-seq analysis. Simultaneously, small RNA was isolated and sequenced. MicroRNAs (miRs) can regulate genes post-transcriptionally and are thought to be important for olfaction gene regulation in the antenna of mosquitoes. Or46 and Or99 are expressed only in unfed females, while Or49 is only expressed post blood meal. A total of 14 IRs, 6 ORs and 6 OBPs are up regulated post feeding, while 8 IRs, 4 ORs and 11 OBPs are down regulated post feeding. A total of 16 new miRs were discovered in the antenna of females. miR-34, miR-79, miR-929, and miR-965 are uniquely expressed in the female antenna; while miR-92b is uniquely expressed in male antenna, while miR-279 is up regulated in the antenna of both males and females. The expression of the miR-275, which is associated with blood meal digestion, increases 3h after blood feeding in the head and thorax and miR-305 displays a similar expression pattern. This is the first study to find miRs that are uniquely expressed in the antenna of mosquitoes. Although there are differences in the expression of olfaction genes in the antenna of 12h old females and 4-day-old females, no changes are observed after blood feeding and similar observations were noted for miRs. Changes may occur only in a specific subset of sensory neurons in the antenna of females and for this reason could not be detected. Olfaction transcripts with long 3’UTR may compete for miRs binding and gene regulation in the antenna of mosquitoes without dramatic changes at the transcript level

Computational analysis of the interaction between transcription factors and the predicted secreted proteome of the yeast Kluyveromyces lactis

<p>Abstract</p> <p>Background</p> <p>Protein secretion is a cell translocation process of major biological and technological significance. The secretion and downstream processing of proteins by recombinant cells is of great commercial interest. The yeast <it>Kluyveromyces lactis </it>is considered a promising host for heterologous protein production. Because yeasts naturally do not secrete as many proteins as filamentous fungi, they can produce secreted recombinant proteins with few contaminants in the medium. An ideal system to address the secretion of a desired protein could be exploited among the native proteins in certain physiological conditions. By applying algorithms to the completed <it>K. lactis </it>genome sequence, such a system could be selected. To this end, we predicted protein subcellular locations and correlated the resulting extracellular secretome with the transcription factors that modulate the cellular response to a particular environmental stimulus.</p> <p>Results</p> <p>To explore the potential <it>Kluyveromyces lactis </it>extracellular secretome, four computational prediction algorithms were applied to 5076 predicted <it>K. lactis </it>proteins from the genome database. SignalP v3 identified 418 proteins with N-terminal signal peptides. From these 418 proteins, the Phobius algorithm predicted that 176 proteins have no transmembrane domains, and the big-PI Predictor identified 150 proteins as having no glycosylphosphatidylinositol (GPI) modification sites. WoLF PSORT predicted that the <it>K. lactis </it>secretome consists of 109 putative proteins, excluding subcellular targeting. The transcription regulators of the putative extracellular proteins were investigated by searching for DNA binding sites in their putative promoters. The conditions to favor expression were obtained by searching Gene Ontology terms and using graph theory.</p> <p>Conclusion</p> <p>A public database of <it>K. lactis </it>secreted proteins and their transcription factors are presented. It consists of 109 ORFs and 23 transcription factors. A graph created from this database shows 134 nodes and 884 edges, suggesting a vast number of relationships to be validated experimentally. Most of the transcription factors are related to responses to stress such as drug, acid and heat resistance, as well as nitrogen limitation, and may be useful for inducing maximal expression of potential extracellular proteins.</p

Qualidade de mudas de tomate em função do substrato e irrigação com efluente de piscicultura.

O objetivo deste trabalho foi avaliar a produção de mudas de tomate em três substratos sob irrigação com efluente de piscicultura. O experimento foi conduzido na horta do Departamento de Ciências Vegetais, na Universidade Federal Rural do Semiárido (UFERSA), Mossoró, RN. O delineamento experimental foi o inteiramente casualizado, em esquema fatorial 3 x 2, com quatro repetições, sendo os tratamentos constituídos pelas combinações tipos de substrato (Plantmax®, composto orgânico e areia lavada) com fonte de água de irrigação (efluente de piscicultura e água de poço). A avaliação foi feita aos 24 dias da germinação através de número de folhas, altura da parte aérea, comprimento da raiz principal, massa seca da parte aérea e massa seca do sistema radicular. O composto orgânico e a areia lavada são os substratos mais indicados para a produção de mudas de tomateiro em função de proporcionar maior número de folhas, altura da planta e massa seca da parte aérea e raiz. O efluente de piscicultura influencia no maior crescimento da raiz, massa seca da parte aérea e raiz quando comparada a água de poço tubular

Origins of high latitude introductions of Aedes aegypti to Nebraska and Utah during 2019

Aedes aegypti (L.), the yellow fever mosquito, is also an important vector of dengue and Zika viruses, and an invasive species in North America. Aedes aegypti inhabits tropical and sub-tropical areas of the world and in North America is primarily distributed throughout the southern US states and Mexico. The northern range of Ae. aegypti is limited by cold winter months and establishment in these areas has been mostly unsuccessful. However, frequent introductions of Ae. aegypti to temperate, non-endemic areas during the warmer months can lead to seasonal activity and disease outbreaks. Two Ae. aegypti incursions were reported in the late summer of 2019 into York, Nebraska and Moab, Utah. These states had no history of established populations of this mosquito and no evidence of previous seasonal activity. We genotyped a subset of individuals from each location at 12 microsatellite loci and ~ 14,000 single nucleotide polymorphic markers to determine their genetic affinities to other populations worldwide and investigate their potential source of introduction. Our results support a single origin for each of the introductions from different sources. Aedes aegypti from Utah likely derived from Tucson, Arizona, or a nearby location. Nebraska specimen results were not as conclusive, but point to an origin from southcentral or southeastern US. In addition to an effective, efficient, and sustainable control of invasive mosquitoes, such as Ae. aegypti, identifying the potential routes of introduction will be key to prevent future incursions and assess their potential health threat based on the ability of the source population to transmit a particular virus and its insecticide resistance profile, which may complicate vector control

Photobiomodulation reduces inflammation but does not influence the hypoxia-inducible factor-1α in pulp tissue of rats after bleaching

Objectives: To evaluate the influence of photobiomodulation with infrared laser (IRL) in the rat pulp tissue after bleaching, considering the immunolabeling of interleukin (IL)-23 and hypoxia-inducible factor (HIF)-1α. Methodology: The right and left molars of forty rats were divided into groups: Control – with placebo gel and Bleached – with 35% hydrogen peroxide (H2O2). Half of the rats received one IRL application on both sides, establishing a split-mouth design, which resulted in 4 groups with 20 hemi-maxillae each: Control, Bleach, IRL, and Bleached-IRL. Rats (n=10) from each group were euthanized, at 2- and 30-days mark, and the pulp tissue was evaluated using inflammation and immunolabeling scores. Wilcoxon and Mann-Whitney statistical tests were performed (p&lt;0.05). Results: At the 2-days mark, the Bleached group had severe inflammation and necrosis in the occlusal thirds of the pulp, and moderate to severe inflammation in cervical third, whereas the Bleached-IRL had mild to moderate inflammation (p&lt;0.05). At the 30-days mark, there was no inflammation, but tertiary dentine formation in the bleached groups. Regarding IL-23, severe immunolabeling was observed in the Bleached group (p&lt;0.05) at the 2-days mark; at the 30-days mark, there was a reduction in immunolabeling, in which the Bleached group had moderate and the Bleached-IRL group had mild immunolabeling (p&gt;0.05). HIF-1α was more evident at the 2-days mark in the Bleached group, without significant difference with the Bleached-IRL (p&gt;0.05). The difference was observed between the bleached and control groups, without immunolabeling (p&lt;0.05); at the 30-days mark, the Bleached group had reduction in HIF-1α immunolabeling, while the Bleached-IRL had an increase; the difference remained between the bleached and the controls groups (p&lt;0.05). Conclusion: Photobiomodulation using IRL minimized the inflammation and IL-23 immunolabeling in the pulp tissue of rats after dental bleaching, but did not influence significantly the HIF-1α immunolabeling

Dietary supplementation with multi-strain formula of probiotics modulates inflammatory and immunological markers in apical periodontitis

Objective: The aim of this study was to evaluate whether probiotics multi-strain formula affects the development of apical periodontitis (AP) induced in rats. Methodology: 16 Wistar rats were divided in two groups (n=8): rats with AP fed with regular diet (Control-C (CG)); rats with AP, fed with regular diet and supplemented with multi-strain formula (one billion colony-forming units (CFU)): GNC Probiotic Complex (PCG) ( Lactobacillus acidophilus, Lactobacillus salivaris, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium bifidum, Bifidobacterium animalis subs. lactis and Streptococcus thermofilus ). AP was induced in the upper and lower first molars by dental pulp exposure to the oral environment. PCG was administered orally through gavage for 30 days during the AP development. After this period the animals were euthanized and the mandibles were removed and processed for histologic analysis, and immunochemical assays for interleukin (IL)-6, IL-10, IL-1β, RANKL, OPG, and TRAP. The Mann–Whitney U test and Student’s t test were performed (P&lt;.05). Results:  The CG showed more intense inflammatory infiltrate than the PCG group (P&lt;.05). IL-1β, IL 6 and RANKL decreased in the PCG group compared with CG (P&lt;.05). The IL-10 level increased in the PCG group (P&lt;.05). The OPG level was similar in both groups (P&gt;.05). The number of mature osteoclasts (TRAP-positive multinucleated cells) was lower in PCG group when compared to the CG (P&lt;.05). Conclusion: Probiotic Complex modulates inflammation and bone resorption in apical periodontitis

Improved reference genome of the arboviral vector Aedes albopictus

Background: The Asian tiger mosquito Aedes albopictus is globally expanding and has become the main vector for human arboviruses in Europe. With limited antiviral drugs and vaccines available, vector control is the primary approach to prevent mosquito-borne diseases. A reliable and accurate DNA sequence of the Ae. albopictus genome is essential to develop new approaches that involve genetic manipulation of mosquitoes. Results: We use long-read sequencing methods and modern scaffolding techniques (PacBio, 10X, and Hi-C) to produce AalbF2, a dramatically improved assembly of the Ae. albopictus genome. AalbF2 reveals widespread viral insertions, novel microRNAs and piRNA clusters, the sex-determining locus, and new immunity genes, and enables genome-wide studies of geographically diverse Ae. albopictus populations and analyses of the developmental and stage-dependent network of expression data. Additionally, we build the first physical map for this species with 75% of the assembled genome anchored to the chromosomes. Conclusion: The AalbF2 genome assembly represents the most up-to-date collective knowledge of the Ae. albopictus genome. These resources represent a foundation to improve understanding of the adaptation potential and the epidemiological relevance of this species and foster the development of innovative control measures

A genotyping array for the globally invasive vector mosquito, Aedes albopictus

Background: Although whole-genome sequencing (WGS) is the preferred genotyping method for most genomic analyses, limitations are often experienced when studying genomes characterized by a high percentage of repetitive elements, high linkage, and recombination deserts. The Asian tiger mosquito (Aedes albopictus), for example, has a genome comprising up to 72% repetitive elements, and therefore we set out to develop a single-nucleotide polymorphism (SNP) chip to be more cost-effective. Aedes albopictus is an invasive species originating from Southeast Asia that has recently spread around the world and is a vector for many human diseases. Developing an accessible genotyping platform is essential in advancing biological control methods and understanding the population dynamics of this pest species, with significant implications for public health. Methods: We designed a SNP chip for Ae. albopictus (Aealbo chip) based on approximately 2.7 million SNPs identified using WGS data from 819 worldwide samples. We validated the chip using laboratory single-pair crosses, comparing technical replicates, and comparing genotypes of samples genotyped by WGS and the SNP chip. We then used the chip for a population genomic analysis of 237 samples from 28 sites in the native range to evaluate its usefulness in describing patterns of genomic variation and tracing the origins of invasions. Results: Probes on the Aealbo chip targeted 175,396 SNPs in coding and non-coding regions across all three chromosomes, with a density of 102 SNPs per 1 Mb window, and at least one SNP in each of the 17,461 protein-coding genes. Overall, 70% of the probes captured the genetic variation. Segregation analysis found that 98% of the SNPs followed expectations of single-copy Mendelian genes. Comparisons with WGS indicated that sites with genotype disagreements were mostly heterozygotes at loci with WGS read depth \u3c 20, while there was near complete agreement with WGS read depths \u3e 20, indicating that the chip more accurately detects heterozygotes than low-coverage WGS. Sample sizes did not affect the accuracy of the SNP chip genotype calls. Ancestry analyses identified four to five genetic clusters in the native range with various levels of admixture. Conclusions: The Aealbo chip is highly accurate, is concordant with genotypes from WGS with high sequence coverage, and may be more accurate than low-coverage WGS. Graphical Abstract: (Figure presented.) © The Author(s) 2024

Influence of the Vehicle on the Tissue Reaction and Biomineralization of Fast Endodontic Cement

Objective: To investigate the tissue response and the biomineralization ability of CER prepared with epoxy resin or water compared to Mineral Trioxide Aggregate (MTA). Material and Methods: Polyethylene tubes containing materials or empty tubes for control were inserted into the subcutaneous tissues of 30 rats. After 7, 15, 30, 60, and 90 days, the rats were killed and the tubes were removed for analysis using hematoxylin-eosin staining, von Kossa staining, and under polarized light. Inflammation was graded through a score system; the thickness of the fibrous capsule was classified as thin or thick; the biomineralization ability was recorded as present or absent. The results were statistically analyzed using the Kruskal-Wallis test (p&lt;0.05). Results: Histologic analysis performed after 7 and 15 days for CER prepared with epoxy resin or water and for MTA showed moderate inflammation and a thick fibrous capsule (p&gt;0.05). After 30, 60, and 90 days, mild inflammation, and a thin fibrous capsule were observed in all groups (p&gt;0.05). Conclusion: All materials had structures positive for von Kossa and birefringent to polarized light. CER epoxy resin showed biocompatibility and biomineralization similar to CER water and MTA

Principal Component and Biplot Analysis in the Agro-industrial Characteristics of Anacardium spp.

The cajuí Anacardium spp., which is similar to the caju Anacardium Ocidentale L., is a species adapted to edaphic-climatic conditions of the biome Cerratinga (Cerrado e Caatinga). Its fruit is composed of one swollen stalk (pedicel) which is formed by nutritional reserves rich in vitamin C and drupe (cashew nut). It is also rich in protein and lipids, but with smaller size. This paper focuses on investigating the applicability of the biplot graphical analysis in the process of selective breeding of cajuí population. The cajuí working European Scientific Journal October 2019 edition Vol.15, No.30 ISSN: 1857 – 7881 (Print) e - ISSN 1857- 7431 22 population in Embrapa Meio Norte comprises of 11 genotypes collected in areas of natural habitat in the state of Piaui. The experiment was designed in randomized complete blocks with two plants per plot and four replications. A graphical analysis (biplot) was used to study the relationships between variables and behavior of the experimental genotypes.&nbsp;This was implemented to principal component analysis based on singular value decomposition biplot. The total variable weight can be predicted from length of peduncle, basal and apical diameter of peduncle, and variables of easy mensuration. Genotypes M40A, M23, M14, and M17 are similar to each other and they have high amounts of brown, apical and basal diameter of the peduncle, total weight, and peduncle length. They are considered as candidates selected for consumption in natura and industrial processing. The graphical analysis (biplot) showed robustness in the presentation of relationships between variables considered and the indication of the selection candidate genotypes in the population studied