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At least 361 records · Page 20

Method of detecting genetic deletions identified with chromosomal abnormalities

Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyzes. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acids probes are typically of a complexity greater tha 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particlularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar ut genetically different diseases, and for many prognostic and diagnostic applications.

60 APPLIED LIFE SCIENCES↗

Genetic barcodes

Herein are described multicolor FISH probe sets termed "genetic barcodes" targeting several cancer or disease-related loci to assess gene rearrangements and copy number changes in tumor cells. Two, three or more different fluorophores are used to detect the genetic barcode sections thus permitting unique labeling and multilocus analysis in individual cell nuclei. Gene specific barcodes can be generated and combined to provide both numerical and structural genetic information for these and other pertinent disease associated genes.

59 BASIC BIOLOGICAL SCIENCES↗

Enhanced Integer Permutation based Genetic Algorithm for Optimization of Tube-Fin Heat Exchanger Circuitry with Splits and Merges

Tube-fin heat exchangers (HXs) are widely used in air-conditioning and heat pump applications. The performance of these heat exchangers is strongly influenced by the refrigerant circuitry. Studies have proved that by optimizing the refrigerant circuitry, the performance of HXs can be significantly improved. In our previous research, an Integer Permutation based Genetic Algorithm (IPGA) was developed to obtain the optimal circuitry designs. Our previous research showed that IPGA demonstrates superior capability to obtain better refrigerant circuitries with lower computational cost than the other methods in literature. And the optimal circuitry designs obtained from IPGA are manufacturable with the available tooling. However, the IPGA developed previously cannot generate designs with splitting and merging of circuits. To remedy this limitation, a new chromosome which can represent circuitry with splitting and merging of circuits is developed. In addition to the six genetic operators implemented previously, two new genetic operators are developed to generate splits and merges. As a result, the enhanced IPGA can explore the solution space more thoroughly than the previous IPGA. A case study using an evaporator from an A-type indoor unit shows that, given the similar capacity improvements obtained from the enhanced IPGA compared with the previous IPGA, the refrigerant pressure drop reduction obtained from the enhanced IPGA is 26.5% compared against 1.0% pressure drop reduction from the previous IPGA. The benchmark of the enhanced IPGA with other methods in literature demonstrates that the enhanced IPGA can generate circuitry designs with performance superior to those obtained from other methods.

LI, Zhenning↗

The Optimal Wavelengths for Light Absorption Spectroscopy Measurements Based on Genetic Algorithm–Particle Swarm Optimization

To select the optimal wavelengths in the light extinction spectroscopy measurement, genetic algorithm–particle swarm optimization (GAPSO) based on genetic algorithm (GA) and particle swarm optimization (PSO) is adopted. The change of the optimal wavelength positions in different feature size parameters and distribution parameters is evaluated. Moreover, the Monte Carlo method based on random probability is used to identify the number of optimal wavelengths, and good inversion effects of the particle size distribution are obtained. The method proved to have the advantage of resisting noise. In order to verify the feasibility of the algorithm, spectra with bands ranging from 200 to 1000 nm are computed. Based on this, the measured data of standard particles are used to verify the algorithm.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Anti-Pdc1p Nanobody as a Genetically Encoded Inhibitor of Ethanol Production Enables Dual Transcriptional and Post-translational Controls of Yeast Fermentations

Microbial fermentation provides a sustainable method of producing valuable chemicals. Adding dynamic control to fermentations can significantly improve titers, but most systems rely on transcriptional controls of metabolic enzymes, leaving existing intracellular enzymes unregulated. This limits the ability of transcriptional controls to switch off metabolic pathways, especially when metabolic enzymes have long half-lives. We developed a two-layer transcriptional/post-translational control system for yeast fermentations. Specifically, the system uses blue light to transcriptionally activate the major pyruvate decarboxylase PDC1, required for cell growth and concomitant ethanol production. Switching to darkness transcriptionally inactivates PDC1 and instead activates the anti-Pdc1p nanobody, NbJRI, to act as a genetically encoded inhibitor of Pdc1p accumulated during the growth phase. This dual transcriptional/post-translational control improves the production of 2,3-BDO and citramalate by up to 100 and 92% compared to using transcriptional controls alone in dynamic two-phase fermentations. Furthermore, this study establishes the NbJRI nanobody as an effective genetically encoded inhibitor of Pdc1p that can enhance the production of pyruvate-derived chemicals.

59 BASIC BIOLOGICAL SCIENCES↗

Contingency rules for pathogen competition and antagonism in a genetically based, plant defense hierarchy

Abstract Plant defense against pathogens includes a range of mechanisms, including, but not limited to, genetic resistance, pathogen‐antagonizing endophytes, and pathogen competitors. The relative importance of each mechanism can be expressed in a hierarchical view of defense. Several recent studies have shown that pathogen antagonism is inconsistently expressed within the plant defense hierarchy. Our hypothesis is that the hierarchy is governed by contingency rules that determine when and where antagonists reduce plant disease severity. Here, we investigated whether pathogen competition influences pathogen antagonism using Populus as a model system. In three independent field experiments, we asked whether competition for leaf mesophyll cells between a Melampsora rust pathogen and a microscopic, eriophyid mite affects rust pathogen antagonism by fungal leaf endophytes. The rust pathogen has an annual, phenological disadvantage in competition with the mite because the rust pathogen must infect its secondary host in spring before infecting Populus . We varied mite–rust competition by utilizing Populus genotypes characterized by differential genetic resistance to the two organisms. We inoculated plants with endophytes and allowed mites and rust to infect plants naturally. Two contingency rules emerged from the three field experiments: (a) Pathogen antagonism by endophytes can be preempted by host genes for resistance that suppress pathogen development, and (b) pathogen antagonism by endophytes can secondarily be preempted by competitive exclusion of the rust by the mite. Synthesis : Our results point to a Populus defense hierarchy with resistance genes on top, followed by pathogen competition, and finally pathogen antagonism by endophytes. We expect these rules will help to explain the variation in pathogen antagonism that is currently attributed to context dependency.

59 BASIC BIOLOGICAL SCIENCES↗

High-yield recombinant bacterial expression of 13 C-, 15 N-labeled, serine-16 phosphorylated, murine amelogenin using a modified third generation genetic code expansion protocol

Amelogenin constitutes ~ 90% of the enamel matrix in the secretory stage of amelogenesis, a still poorly understood process that results in the formation of the hardest and most mineralized tissue in vertebrates – enamel. Most biophysical research with amelogenin uses recombinant protein expressed in Escherichia coli. In addition to providing copious amounts of protein, recombinant expression allows 13 C- and 15 N-labelling for detailed structural studies using NMR spectroscopy. However, native amelogenin is phosphorylated at one position, Ser-16 in murine amelogenin, and a lingering question overshadowing decades of enamel research with recombinant amelogenin is the effect of phosphorylation on its biophysical properties. Using a modified genetic code expansion protocol we have expressed and purified uniformly 13 C-, 15 N-labeled murine amelogenin (pS16M179) with ~95% of the protein being correctly phosphorylated. Homogeneous phosphorylation was achieved using commercially available, enriched, 13 C-, 15 N-labeled media and protein expression was induced with isopropyl β-D-1-thiogalactopyranoside at 310 K. Phosphoserine incorporation was verified from one-dimensional 31P NMR spectra, comparison of 1 H- 15 N HSQC spectra, Phos-tag SDS PAGE, and mass spectrometry. Phosphorus-31 NMR spectra for pS16M179 under conditions known to trigger amelogenin self-assembly into nanospheres confirm nanosphere models with buried N-termini. Lambda phosphatase treatment of these nanospheres results in the dephosphorylation of pS16M179, confirming that smaller oligomers and monomers with exposed N-termini are in equilibrium with nanospheres. Such 13 C-, 15 N-labeling of amelogenin with accurately encoded phosphoserine incorporation will accelerate biomineralization research to understand amelogenesis and stimulate the expanded use of genetic code expansion protocols to introduce phosphorylated amino acids into proteins.

31P-NMR↗

A genetic meta-algorithm-assisted inversion approach: hydrogeological study for the determination of volumetric rock properties and matrix and fluid parameters in unsaturated formations

An evolutionary inversion approach is suggested for the interpretation of nuclear and resistivity logs measured by direct-push tools in shallow unsaturated sediments. The efficiency of formation evaluation is improved by estimating simultaneously (1) the petrophysical properties that vary rapidly along a drill hole with depth and (2) the zone parameters that can be treated as constant, in one inversion procedure. In the workflow, the fractional volumes of water, air, matrix and clay are estimated in adjacent depths by linearized inversion, whereas the clay and matrix properties are updated using a float-encoded genetic meta-algorithm. The proposed inversion method provides an objective estimate of the zone parameters that appear in the tool response equations applied to solve the forward problem, which can significantly increase the reliability of the petrophysical model as opposed to setting these parameters arbitrarily. The global optimization meta-algorithm not only assures the best fit between the measured and calculated data but also gives a reliable solution, practically independent of the initial model, as laboratory data are unnecessary in the inversion procedure. The feasibility test uses engineering geophysical sounding logs observed in an unsaturated loessy-sandy formation in Hungary. The multi-borehole extension of the inversion technique is developed to determine the petrophysical properties and their estimation errors along a profile of drill holes. The genetic meta-algorithmic inversion method is recommended for hydrogeophysical logging applications of various kinds to automatically extract the volumetric ratios of rock and fluid constituents as well as the most important zone parameters in a reliable inversion procedure.

58 GEOSCIENCES↗

Evaluation of phenotype stability and ecological risk of a genetically engineered alga in open pond production

Genetically engineered (GE) algae offer the promise of producing food, fuel, and other valuable products with reduced requirements for land and fresh water. While the gains in productivity measured in GE terrestrial crops are predicted to be mirrored in GE algae, the stability of phenotypes and ecological risks posed by GE algae in large-scale outdoor cultivation remain unknown. In this paper, we describe the first US Environmental Protection Agency (EPA)-sanctioned experiment aimed at understanding how GE algae perform in outdoor cultivation. Acutodesmus dimorphus was genetically engineered by the addition of two genes, one for enhanced fatty acid biosynthesis, and one for recombinant green fluorescence protein (GFP) expression; both the genes and their associated phenotypes were maintained during fifty days of outdoor cultivation. We also observed that while the GE algae dispersed from the cultivation ponds, colonization of the trap ponds by the GE strain declined rapidly with increasing distance from the source cultivation ponds. In contrast, many species of indigenous algae were found in every trap pond within a few days of starting the experiment. When inoculated in water from five local lakes, the GE algae's effect on biodiversity, species composition, and biomass of native algae was indiscernible from those of the wild-type (wt) progenitor algae, and neither the GE nor wt algae were able to outcompete native strains. We conclude that GE algae can be successfully cultivated outdoors while maintaining GE traits, and that for the specific GE algal strain tested here they did not outcompete or adversely impact native algae populations when grown in water taken from local lakes. Finally, this study provides an initial evaluation of GE algae in outdoor cultivation and a framework to evaluate GE algae risks associated with outdoor GE algae production.

09 BIOMASS FUELS↗

Successful post-translocation reproduction and genetic integration of eastern box turtles

Translocation is a conservation tool increasingly used in the recovery of at-risk species, including turtles, which are one of the world's most imperiled taxa. Post-release monitoring is essential to determine the outcomes of a given intervention and inform future efforts. However, monitoring typically focuses on post-release survival and spatial ecology whereas few studies assess the genetic and demographic outcomes. The eastern box turtle (Terrapene carolina carolina) is in decline throughout its range and is increasingly likely to be subject to translocations, including efforts to repatriate animals confiscated from the illegal wildlife trade. In 2019–2021, we translocated two groups of box turtles to the Savannah River Site in South Carolina, USA, including confiscated turtles (n = 208) and surrendered long-term captive turtles (LTC; n = 35). In 2022, we monitored a subset of confiscated (n = 12), LTC (n = 15), and sympatric resident (n = 8) females for reproductive output and genotyped their offspring and candidate sires to assign parentage. We found that all groups of females produced eggs at a similar rate and produced viable offspring but that the most recently translocated group (LTCs) displayed lower hatching success. Parentage assignment revealed that all groups sired offspring and mated with each other. Furthermore, our results broadly indicate that confiscated and LTC box turtles can successfully reproduce and genetically integrate following their release into wild populations, and that translocation may serve as a valuable tool for local population recovery.

59 BASIC BIOLOGICAL SCIENCES↗

Cross-kingdom expression of synthetic genetic elements promotes discovery of metabolites in the human microbiome

Small molecules encoded by biosynthetic pathways mediate cross-species interactions and harbor untapped potential, which has provided valuable compounds for medicine and biotechnology. Since studying biosynthetic gene clusters in their native context is often difficult, alternative efforts rely on heterologous expression, which is limited by host-specific metabolic capacity and regulation. Here, in this work, we describe a computational-experimental technology to redesign genes and their regulatory regions with hybrid elements for cross-species expression in Gram-negative and -positive bacteria and eukaryotes, decoupling biosynthetic capacity from host-range constraints to activate silenced pathways. These synthetic genetic elements enabled the discovery of a class of microbiome-derived nucleotide metabolites—tyrocitabines—from Lactobacillus iners. Tyrocitabines feature a remarkable orthoester-phosphate, inhibit translational activity, and invoke unexpected biosynthetic machinery, including a class of “Amadori synthases” and “abortive” tRNA synthetases. Our approach establishes a general strategy for the redesign, expression, mobilization, and characterization of genetic elements in diverse organisms and communities.

59 BASIC BIOLOGICAL SCIENCES↗

Inverse design of hypoeutectoid pearlite steel microstructures using a deep learning and genetic algorithm optimization framework

Goal-oriented microstructure design in metallic materials is a challenging task due to complex structure-property relationships. Traditional experimental and computational approaches are time-intensive and economically inefficient, limiting their applicability for large-scale design space exploration. Here, in this work, we propose an end-to-end framework that integrates deep learning models with genetic optimization to design microstructures with targeted mechanical properties. Deep learning models enable accurate forward design, while their integration with genetic optimization enables efficient inverse design within a few hours, compared to days or weeks using conventional finite element simulations. The framework combines experimental characterization and finite element modeling to analyze the influence of microstructural features on the mechanical behavior of hypoeutectoid steels. Data from both experiments and simulations are used to train the deep learning models. To demonstrate its effectiveness, we apply the framework to 0.63% carbon steel with proeutectoid ferrite and pearlite phases, commonly used in industrial applications. In this study, 2D microstructures were used for modeling, selected primarily for computational efficiency and to establish proof of concept. The framework successfully optimizes microstructures for targeted yield strength, ultimate strength, and stress concentration factors while significantly reducing computational time. Beyond hypoeutectoid steels, this scalable framework can be extended to other material systems and integrated with additive manufacturing, offering an efficient approach for accelerating microstructure design for specific engineering applications.

ConvLSTM↗

Arsenic methylation capacity in relation to nutrient intake and genetic polymorphisms in one-carbon metabolism

Highlights: • FOLH1 CC, MTR GG, and MTHFD1 GG carriers had better iAs methylation capacity. • Vitamin B12 interacted with FOLH1 c.223 T > C on iAs metabolism. • One-carbon nutrients and genetic variants may jointly influence iAs metabolism. Nutrients and genetic polymorphisms participating in one-carbon metabolism may explain interindividual differences in inorganic arsenic (iAs) methylation capacity, which in turn may account for variations in susceptibility to iAs-induced diseases.

54 ENVIRONMENTAL SCIENCES↗

Machine learning-guided discovery of polymer membranes for CO 2 separation with genetic algorithm

Designing polymer membranes with high gas permeability and selectivity is a difficult multi-task constrained problem due to the trade-off between these two properties. In this work, we present a machine learning (ML) driven genetic algorithm to tackle the design problem of polymer membranes for CO 2 separation from N 2 and O 2 . Using literature data of permeability for three gases, we constructed multiple ML models with different fingerprinting featurization schemes to predict gas permeabilities. Then, we employed a genetic algorithm to design new polymers and evaluated their performance using our ML models. We were able to identify new polymer membranes that are promising for both CO 2 /N 2 and CO 2 /O 2 separations. Further, the top discovered polymers are predicted to have high glass transition temperatures. Similarly, the pyridine functionality was found in ≈20% of the predicted polymers. This framework can be used to design polymers for any application involving constrained optimization. Finally, we outlined the challenges and opportunities with using ML guided data-driven inverse design of polymers.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Development of a reverse genetics system to generate a recombinant Ebola virus Makona expressing a green fluorescent protein

Previous studies have demonstrated the potential application of reverse genetics technology in studying a broad range of aspects of viral biology, including gene regulation, protein function, cell entry, and pathogenesis. Here, we describe a highly efficient reverse genetics system used to generate recombinant Ebola virus (EBOV) based on a recent isolate from a human patient infected during the 2014–2015 outbreak in Western Africa. We also rescued a recombinant EBOV expressing a fluorescent reporter protein from a cleaved VP40 protein fusion. Using this virus and an inexpensive method to quantitate the expression of the foreign gene, we demonstrate its potential usefulness as a tool for screening antiviral compounds and measuring neutralizing antibodies. - Highlights: • Recombinant Ebola virus (EBOV) derived from Makona variant was rescued. • New protocol for viral rescue allows 100% efficiency. • Modified EBOV expresses a green fluorescent protein from a VP40-fused protein. • Modified EBOV was tested as tool to screen antiviral compounds and measure neutralizing antibodies.

60 APPLIED LIFE SCIENCES↗

Human adipose-derived mesenchymal stromal cells from face and abdomen undergo replicative senescence and loss of genetic integrity after long-term culture

Body fat depots are heterogeneous concerning their embryonic origin, structure, exposure to environmental stressors, and availability. Thus, investigating adipose-derived mesenchymal stromal cells (ASCs) from different sources is essential to standardization for future therapies. In vitro amplification is also critical because it may predispose cell senescence and mutations, reducing regenerative properties and safety. Here, we evaluated long-term culture of human facial ASCs (fASCs) and abdominal ASCs (aASCs) and showed that both met the criteria for MSCs characterization but presented differences in their immunophenotypic profile, and differentiation and clonogenic potentials. The abdominal tissue yielded more ASCs, and these had higher proliferative potential, but facial cells displayed fewer mitotic errors at higher passages. However, both cell types reduced clonal efficiency over time and entered replicative senescence around P12, as evaluated by progressive morphological alterations, reduced proliferative capacity, and SA-β-galactosidase expression. Loss of genetic integrity was detected by a higher proportion of cells showing nuclear alterations and γ-H2AX expression. Our findings indicate that the source of ASCs can substantially influence their phenotype and therefore should be carefully considered in future cell therapies, avoiding, however, long-term culture to ensure genetic stability.

60 APPLIED LIFE SCIENCES↗

Genetically engineered mouse models of esophageal cancer

Esophageal cancer is the most common cause of cancer-related death worldwide with a diverse geographical distribution, poor prognosis, and diagnosis in advanced stages of the disease. Identification of the mechanisms involved in esophageal cancer development is evaluative to improve outcomes for patients. Genetically engineered mouse models (GEMMs) of cancer provide the physiologic, molecular, and histologic features of the human tumors to determine the pathogenesis and treatments for cancer, hence exhibiting a source of tremendous potential for oncology research. The advancement of cancer modeling in mice has improved to the extent that researchers can observe and manipulate the disease process in a specific manner. Despite the significant differences between mice and humans, mice can be great models for human oncology researches due to similarities between them at the molecular and physiological levels. Due to most of the existing esophageal cancer GEMMs do not propose an ideal system for pathogenesis of the disease, genetic risks, and microenvironment exposure, so identification of challenges in GEM modeling and well-developed technologies are required to obtain the most value for patients. In this review, we describe the biology of human and mouse, followed by the exciting esophageal cancer mouse models with a discussion of applicability and challenges of these models for generating new GEMMs in future studies.

60 APPLIED LIFE SCIENCES↗

System-Level Optimization to Improve Biofuel Potential via Genetic Engineering and Hydrothermal Liquefaction

The economic success of biofuels and bioproducts depends on system-level optimization including biomass production and conversion. Hydrothermal liquefaction (HTL) can convert wet biomass such as microalgae into a biofuel intermediate (BFI) under elevated temperatures and pressure. An understanding of the impacts of biomass composition on BFI yield and quality can inform genetic engineering strategies in the improvement of biochemical composition for biofuel production. In this work, wild type cyanobacterium Synechocystis sp. PCC 6803 biomass was doped with various common cellular storage compounds in lab-scale HTL experiments. Doping with glycogen or polyhydroxybutyrate (PHB) significantly reduced BFI yields, while doping with triglycerides (TAG) or medium chain-length polyhydroxyalkanoate (mcl-PHA) increased BFI yield and quality. In light of these observations, a genetically engineered Synechocystis strain deficient in glycogen biosynthesis was cultivated to produce biomass for HTL, leading to a 17% increase in BFI yield. In addition, we built a multiphase component additivity (MCA) model that can predict BFI yield and quality with PHAs in the biomass. This work demonstrates an effective strategy to integrate strain development with downstream biomass conversion to maximize biofuel yield, with lessons applicable to microalgae as well as other biomass.

09 BIOMASS FUELS↗