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[This corrects the article DOI: 10.17912/micropub.biology.000693.].

Human cancer cell lines are widely used in artificial culture systems to study cancer biology due to their immortalization, enabling extended culturing compared to primary cells. However, genomic instability can rapidly alter cellular behavior, raising questions about the transcriptome stability for controlled studies. We cultivated HCT-116 cells over 25 passages (~16 weeks), and observed a stable transcriptome up to passage 20, with only minimal differential gene expression. Above passage 20, genes corresponding to cell proliferation and cell death were upregulated suggesting a change in general cellular regulation and underlining defined cell culture time periods for stable and reproducible experimental conditions.

Congenital heart defects affect females and males differently. Several congenital heart defects arise during the formation of the heart tube, suggesting that heart tube morphogenesis may differ between females and males. We investigated if the fruit fly Drosophila melanogaster displays sexual dimorphisms in the cellular mechanisms of heart tube formation. Quantitative microscopy revealed no differences between females and males in the migration of cardiac progenitors to form the heart tube. Our results suggest that Drosophila do not display sexual dimorphisms in early cardiac development, and support the omission of sex as an experimental variable when investigating Drosophila heart tube morphogenesis.

Lakes often exist in two alternative stable states: clear waters dominated by submerged aquatic vegetation and turbid states with an abundance of phytoplankton. Management decisions must balance the value of primary producers with recreational use. We monitored the phytoplankton community surrounding removal of invasive Potamogeton crispus , a submerged aquatic perennial. Focus was given to presence of cyanobacteria, which may thrive in the absence of macrophytes by virtue of phosphorus that would otherwise be stored in plant biomass. Results demonstrate that macrophyte removal can lead to an increase in cyanobacteria dominance, with sustained impacts after hand removal.

The CYP307 enzymes are necessary for the synthesis of the arthropod ecdysteroid moulting hormones, but show a notably elevated duplication and loss rate compared to other enzymes acting in the same pathway. We demonstrate that Drosophila melanogaster Cyp307a1 null homozygotes can be rescued by ubiquitous expression of Drosophila hydei Cyp307a3 and Aedes aegypti Cyp307b transgenes, although amino acid identity between D . melanogaster CYP307A1 and A . aegypti CYP307B is less than 35%. This evidence of functional conservation across even distantly-related members of the Cyp307 family provides context for interpreting the evolutionary forces driving Cyp307 copy number variation.

Senescent cell (SC) accumulation theoretically causes aging-related tissue malfunction. We examined SC burden in short-lived and long-lived Nothobranchius furzeri fish strains, and fasting's effects on SCs. Despite different lifespans, SC accumulation rates didn't correspond between strains. The long-lived strain showed sex-based lifespan differences without SC burden differences. A 14-day fast reduced SC burden in short-lived fish, but this effect was temporary. Fasting extended median lifespan without affecting maximum lifespan, suggesting transient healthspan improvement. We conclude that saβgal-assessed SC burden doesn't necessarily correspond with lifespan between strains but may within strains. Our findings confirm intermittent fasting improves health and median lifespan.

Brain tumors remain among the most aggressive cancers due to their ability to adapt to microenvironmental stress to sustain malignancy and resist therapy. We propose Drosophila lethal(3) tumorous brain [ l(3)tb ] mutant, which develops rapidly expanding brain tumors, as a genetically tractable in vivo model to study stress-adaptive tumor growth. The progressive brain enlargement in l(3)tb/l(3)tb larvae is accompanied by robust tumor size-dependent induction of Hsp70, a molecular chaperone linked to poor glioma prognosis. These findings position the l(3)tb model as a powerful platform for evaluation of stress tolerance in brain tumors.

The evolutionarily conserved RNA-binding protein Muscleblind can function as both a splicing regulator in the nucleus and a mRNA stabilizer in the cytosol. C. elegans mbl-1/ Muscleblind undergoes alternative splicing to generate long and short isoforms that contain one or two KR motifs needed for nuclear localization. We generate three alleles that express MBL-1 proteins with two, one, or no KR motifs and find that the proteins with two KR motifs are restricted in the nucleus and could not promote neurite growth in a sensitized background. Surprisingly, proteins with one or no KR motifs are located in both cytoplasm and nucleus.

Developing a gene model for the Glutathione S transferase O3 ortholog ( GstO3 ) in the ASM1815212v1 Genome Assembly (GenBank Accession: GCA_018152125.1) of Drosophila dunni . This ortholog was characterized as part of a developing dataset for a comparative study of detoxification gene family evolution in the immigrans - tripunctata radiation of the genus Drosophila using an adapted Genomics Education Partnership gene annotation protocol for Course-based Undergraduate Research Experiences.

Neurofibromatosis type 1 (NF1) and NF2 -related Schwannomatosis ( NF2 -SWN) are both inherited syndromes characterized by Schwann cell tumors. NF1 tumors harbor activated Ras/MEK/ERK, while NF2 -SWN tumors harbor activated mechanosignaling pathways, including Hippo/YAP-TAZ/TEAD. To test combinatorial strategies in tumor cell lines, we first screened a new-generation TEAD inhibitor, VT103, against 123 drugs and then validated the hits with pairwise titrations. VT103 consistently synergized with inhibitors of MEK (trametinib and selumetinib), SHP2 (TNO155) and mTOR (everolimus). The highest synergy ZIP score, calculated using SynergyFinder, was ~65 in the NF2 -SWN cell line SC4, with lower magnitudes in an NF1 cell line.

Mei-P26 is associated with cell-fate regulation in Drosophila melanogaster . In prior work, offspring of Western diet-fed fathers exhibited hyperphagia accompanied by increased brain Mei-P26. Because feeding is regulated by insulin/ILP signaling, we tested whether altering mei-p26 expression in insulin-producing cells affects behavioral and metabolic outcomes. Here we show that overexpression of mei-p26 in ILP2 neurons reduced feeding (FLIC), consumption (ConEx), locomotor activity, and glucose levels by 30-50%, whereas mei-p26 knockdown produced opposite effects. These effects were consistent across sexes, identifying Mei-P26 as a potential regulator within insulin-producing cells associated with bidirectional changes in adult behavioral and metabolic readouts.

Minoxidil (MX) is a common treatment for androgenetic alopecia (AGA). While there are many proposed mechanisms through which MX may increase hair growth, a clear connection to sex-hormone pathways has yet to be established. With recent evidence suggesting that MX may directly bind to Androgen Receptor (AR) and act as an anti-androgen, we investigated whether MX might also exhibit estrogenic activity. Estrogen-dependent cell lines, tryptophan emission, and computational docking were used to probe the possible Estrogen Receptor α (ERα) agonist activity of MX. Preliminary results suggest MX may be a partial agonist of ERα.

Escherichia albertii is a zoonotic pathogen frequently misidentified as diarrheagenic E. coli due to shared phenotypic and genetic traits, yet functional genomic studies in this species have been limited by the inefficiency of traditional genetic tools. To address this, we developed a bipartite CRISPR/Cas9-assisted lambda red (𝛌-Red) recombineering system for efficient, markerless genome editing in E. albertii . As proof of principle, we targeted the nonessential lacZ gene for negative selection by using Cas9 to generate lethal double stranded breaks in unedited cells. Using a recombinogenic single-stranded DNA (ssDNA) oligonucleotide to introduce a premature stop codon and an NheI restriction site, we achieved a recombination efficiency of 53%. Extending the induction time of the 𝛌-Red recombinase genes enhanced recombineering efficiency to 80%. This optimized CRISPR-assisted platform, the first reported application of its kind in E. albertii, enables the rapid generation of scarless mutations and provides a robust tool for the systematic analysis of E. albertii virulence factors and regulatory networks.

Hairy/Enhancer of Split (HES) proteins are critical for animal development and for the regulation of human diseases. The Caenorhabditis elegans genome encodes six HES orthologs, including HLH-29 . We used Nanopore sequencing on the MinION platform to define the location and organization of an integrated transgenic array expressing green fluorescent protein driven by the hlh-29 promoter. The array, ardIS501 , is at least 188.5 kb long, is inserted into Chromosome III of Bristol N2 , and contains at least 26 copies of hlh-29 p::gfp and 11 copies of rol-6 ( su1006 ) . The coding sequences Y46E12A.2 .1 and Y46E12A.5 .1 are deleted in this strain, with no observable phenotypes.

Caenorhabditis elegans is an attractive, genetically tractable model organism widely used to investigate morphological and locomotor responses to environmental and dietary factors. This study examined the effects of the natural alternative sweeteners, Stevia in the Raw®, Truvia®, Lakanto®, and Monkfruit in the Raw®. Worms were exposed to each sweetener through their growth media, and behavioral (crawling and swimming speed, wavelength, and activity) and morphological (length, width, and area) parameters were quantified using WormLab. The objective was to evaluate the effects on development and behavior using natural alternative sweeteners relative to conventional granulated white sugar when administered in equivalent amounts.

The methyltransferase-like 5 (METTL5) protein has been shown to catalyze m6A deposition on 18S ribosomal RNA. However, whether it also methylates mRNAs remains unclear. To address this, we employed direct RNA sequencing (ONT) for m6A detection on native mRNA molecules. We first validated the quantitative detection of m6A by ONT by treating mESCs with a METTL3 inhibitor. We then compared methylation levels of m6A sites between Mettl5 -KO and WT mESCs. Our analysis provides no compelling evidence for METTL5 mediated mRNA methylation in vivo, indicating that its catalytic activity is restricted to rRNA.

11-aminoundecanoic acid is a photochemical degradation product of the synthetic plastic nylon 11. With increasing plastic waste in the environment, understanding the fates of nylon 11 degradation product is important. Previously, a bacterial strain with an ability to metabolically degrade 11-aminoundecanoic acid was isolated, but the enzymes that are involved in the metabolism of 11-aminoundecanoic acid has yet to be determined. In this work, we developed an enzyme-coupled assay to study transaminase activity with 11-aminoundecanoic acid and pyruvate as co-substrates. Using this method, transaminase activity was successfully detected in the cell lysate of Pseudomonas strain JG-B, grown on 11-aminoundecanoic acid.

The GeneSwitch system (GS) is a genetic reagent for pharmacologically inducible gene expression in Drosophila using the Gal4-UAS binary expression system. We have identified that the pan-glial repo -GS-Gal4 line uniquely causes significant baseline sleep reduction during both day and night in the presence of the inducing agent RU-486 even in the absence of effector transgene expression. Careful experimental design to permit detection of small sleep changes that may be obscured by the large sleep reduction in the presence of RU-486 is required when using the repo -GS-Gal4 line for behavioral studies.

Phage Bouclier has a 19,912bp genome with 29 protein coding genes and infects Arthrobacter globiformis B-2979. Transmission electron microscopy visualizes Bouclier as a podovirus. This is confirmed by identification of genes encoding structural proteins found in other podoviriruses. The portal, upper collar, tail knob, and major capsid proteins can be modeled using AlphaFold3 into macromolecular structures. These structural components have high confidence matches in ChimeraX to Bacillus phage Φ29 that has been resolved by cryoEM. Bouclier contains a defined lysis cassette including two genes encoding Lysin A proteins and three putative transmembrane proteins that may be involved in host lysis.