Later Stages Of Development Research Articles

Diversifying selection in aly, bam, dj and topi genes: Association between Rapid divergence and Hybrid male sterility in Drosophila bipectinata species complex

The diversity of millions of species depends on the multiplication of species and speciation, in which reproductive isolation evolves, such as hybrid sterility: a puzzle that still remains unknown. Crosses between closely related species results in sterile hybrid males, the perturbances of spermatogenesis in the F1 hybrid males usually affect the later stages of development, arresting at the sperm individualization stage, which could be due to the rapid interspecies divergence of reproductive genes experiencing strong selective constraints between species. We annotated and analyzed the molecular evolution of 6 key spermatogenesis genes playing an important role in the sperm developmental pathway in 15 strains of Drosophila bipectinata species subcomplex, where the interspecies crosses are known to produce sterile hybrid males. We analyzed nucleotide divergence and characterized the patterns of sequence polymorphism in the protein-coding regions to validate or reject the nature of adaptive divergence using its phylogenetic histories. Except for spermatocyte arrest (sa) and twine, all 4 genes - always early(aly), bag of marbles(bam), don juan (dj) and matotopetli (topi) showed signatures of positive selection with more nonsynonymous to synonymous fixed differences between species. However, departure from neutrality in polymorphism analysis was only significant for topi, twine and aly. Consistent with our analysis proving rapid divergence of don juan(dj) and topi, hybrids involving the three species Drosophila bipectinata, Drosophila parabipectinata and Drosophila malerkotliana, have found immotile sperm and unindividualized spermatids. Bag of marbles(bam) and aly showed rapid divergence coherent to the aspermic testes abnormality found in the hybrid males from crosses involving the Drosophila pseudoananassae with the other three species. We propose that faster evolution of spermatogenesis genes due to a yet unknown underlying mechanism acting upon the germ cell line developmental pathway of these four species might be operating as a perpetual dynamo for interspecies divergence and hybrid male sterility. Our study is the first attempt to understand the molecular genetic basis of post-zygotic reproductive isolation in bipectinata species complex and lays a foundation for future detailed exploration to better understand the mechanisms involved.

Macro and microscopic maturation stage key of green crab (Carcinus maenas, Linnaeus 1758): Reproductive cycle and differences among estuarine systems

Gonadal histology is a helpful tool to validate species' macroscopic maturity stages in fisheries biology. Regardless of the many studies on Carcinus maenas, there are still few concerning gonads tissue histology and description of maturity stages. In Portugal, information regarding this species' biology to help support the regulation of crab fisheries, including the minimum landing size, has not yet been defined. In this work, the macro and microscopic characteristics of the reproductive cycle of C. maenas, oogenesis, and spermatogenesis development stages along the Portuguese coast are described, and a new macroscopic scale based on the histological analyses is suggested. During 2019 and 2020, adult C. maenas (both males and females) were collected from the Ria de Aveiro estuary, Rio Sado estuary, and Ria Formosa lagoon, respectively, North, Center, and South Portugal. No-significant differences (P > 0.05) were observed in carapace width and individual weight between all systems. Significant differences (P < 0.05) were observed in gonad weight, gonadosomatic index, and Fulton's condition index between the Rio Sado estuary and Ria Formosa lagoon. Furthermore, significant differences (P < 0.05) in the oocyte diameter between the northern and southernmost locations suggest a geographic variation related to the local environmental conditions of each system. Principal component analysis (PCA) revealed maturity stage and carapace width an association, and a similar morphometry between Ria de Aveiro and Ria Formosa. Temperature was correlated to both maturity stage and carapace width in Ria de Aveiro and Ria Formosa. From the histological analysis and based on microscopic criteria, it was observed that females previously classified within the late macroscopic development stage 2 should be considered mature, so a classification change in the current ovary's developmental stage is proposed. These findings can allow fisheries researchers to reclassify the estimations of maturity ogives and help support the regulation of this species' fishery. The proposed macroscopic scale was validated by histological analyses and can be used elsewhere.

SOME MORPHOLOGICAL FEATURES OF THE SYSTEM OF NOUNS IN THE ODESSA COPY OF THE NEW BULGARIAN TRANSLATION OF THE “SMALL CATECHISMOS” BY THEODOR THE STUDITE

The purpose of the article is to consider the morphological peculiarities of the system of the nouns in New Bulgarian translation of the “Catechismos” written by Theodore the Studite, which is a part of the manuscripts no. 1/154 kept in Odessa National Scientific Library. The subject of the research is the morphological specifics of nouns in Odessa copy of the “Catechismos” dating from the 18th. The morphological peculiarities of nouns is considered in the context of the formation of a linguistic norm, which allowed the combination with different intensity of linguistic means of several language systems functioning at the time (traditional Middle Bulgarian written language, Church Slavonic Eastern recensions, and vernacular language form). The analysis proposed in this paper presents the extensive system of cases, which does not reflect the real vernacular Bulgarian speech in the 18th; the specifics of the functioning of the gramemes of the case paradigm of masculine, feminine and neuter nouns in the singular and plural forms is analyzed. Usage case endings mistakes, which indicates their artificial nature, are considered. The lack of article of nominal parts of speech is noted; the predominance of compound declension form of the adjectives and participles over short forms is revealed; the relatively high frequency of use of active present participles is registered. The results of the study make it possible to outline some probable factors that determine the writer’s preference for using the linguistic tools of the so-called “bookish”, “traditional”, “archaic” writing systems. An another reason which to some extent explains he usage of case inflections in the text of this relatively late stage of the historical development of the Bulgarian language might be the use of East Slavic copies of the Studite’s sermons by the scriber. Еhe comparison of “Catechismos” copies of South and East Slavic origin is necessary for verification of this assumption, in which we see prospects for further research.

Single-cell RNA sequencing of murine hearts for studying the development of the cardiac conduction system

The development of the cardiac conduction system (CCS) is essential for correct heart function. However, critical details on the cell types populating the CCS in the mammalian heart during the development remain to be resolved. Using single-cell RNA sequencing, we generated a large dataset of transcriptomes of ~0.5 million individual cells isolated from murine hearts at six successive developmental corresponding to the early, middle and late stages of heart development. The dataset provides a powerful library for studying the development of the heart’s CCS and other cardiac components. Our initial analysis identified distinct cell types between 20 to 26 cell types across different stages, of which ten are involved in forming the CCS. Our dataset allows researchers to reuse the datasets for data mining and a wide range of analyses. Collectively, our data add valuable transcriptomic resources for further study of cardiac development, such as gene expression, transcriptional regulation and functional gene activity in developing hearts, particularly the CCS.

Development of associational fiber tracts in fetal human brain: a cadaveric laboratory investigation.

The advent of diffusion tensor imaging (DTI) in addition to cadaveric brain dissection allowed a comprehensive description of an adult human brain. Nonetheless, the knowledge of the development of the internal architecture of the brain is mostly incomplete. Our study aimed to provide a description of the anatomical variations of the major associational bundles, among fetal and early post-natal periods. Seventeen formalin-fixed fetal human brains were enrolled for sulci analysis, and 13 specimens were dissected under the operating microscope, using Klingler's technique. Although fronto-temporal connections could be observed in all stages of development, a distinction between the uncinate fascicle, and the inferior fronto-occipital fascicle was clear starting from the early preterm period (25-35 post-conceptional week). Similarly, we were consistently able to isolate the periatrial white matter that forms the sagittal stratum (SS), with no clear distinction among SS layers. Arcuate fascicle and superior longitudinal fascicle were isolated only at the late stage of development without a reliable description of their entire course. The results of our study demonstrated that, although white matter is mostly unmyelinated among fetal human brains, cadaveric dissection can be performed with consistent results. Furthermore, the stepwise development of the associational fiber tracts strengthens the hypothesis that anatomy and function run in parallel, and higher is the cognitive functions subserved by an anatomical structure, later the development of the fascicle. Further histological-anatomical-DWI investigations are required to appraise and explore this topic.

Characterization of the Abracl-Expressing Cell Populations in the Embryonic Mammalian Telencephalon.

Abracl (ABRA C-terminal-like protein) is a small, non-typical winged-helix protein that shares similarity with the C-terminal domain of the protein ABRA (Actin-Binding Rho-Activating protein). The role of Abracl in the cell remains elusive, although in cancer cells, it has been implicated in proliferation, migration and actin dynamics. Our previous study showed that Abracl mRNA was expressed in the dividing cells of the subpallial subventricular zone (SVZ), in the developing cortical plate (CP), and in the diencephalic SVZ; however, the molecular identities of the Abracl-expressing cell populations were not defined in that work. In this study, we use double immunofluorescence to characterize the expression of Abracl on sections of embryonic murine (E11.5-E18.5) and feline (E30/31-E33/34) telencephalon; to this end, we use a battery of well-known molecular markers of cycling (Ki67, Ascl1, Dlx2) or post-mitotic (Tubb3, Gad65/67, Lhx6 and Tbr1) cells. Our experiments show that Abracl protein has, compared to the mRNA, a broader expression domain, including, apart from proliferating cells of the subpallial and diencephalic SVZ, post-mitotic cells occupying the subpallial and pallial mantle (including the CP), as well as subpallial-derived migrating interneurons. Interestingly, in late embryonic developmental stages, Abracl was also transiently detected in major telencephalic fiber tracts.

CUX1-related neurodevelopmental disorder: deep insights into phenotype-genotype spectrum and underlying pathology

Heterozygous, pathogenic CUX1 variants are associated with global developmental delay or intellectual disability. This study delineates the clinical presentation in an extended cohort and investigates the molecular mechanism underlying the disorder in a Cux1+/− mouse model. Through international collaboration, we assembled the phenotypic and molecular information for 34 individuals (23 unpublished individuals). We analyze brain CUX1 expression and susceptibility to epilepsy in Cux1+/− mice. We describe 34 individuals, from which 30 were unrelated, with 26 different null and four missense variants. The leading symptoms were mild to moderate delayed speech and motor development and borderline to moderate intellectual disability. Additional symptoms were muscular hypotonia, seizures, joint laxity, and abnormalities of the forehead. In Cux1+/− mice, we found delayed growth, histologically normal brains, and increased susceptibility to seizures. In Cux1+/− brains, the expression of Cux1 transcripts was half of WT animals. Expression of CUX1 proteins was reduced, although in early postnatal animals significantly more than in adults. In summary, disease-causing CUX1 variants result in a non-syndromic phenotype of developmental delay and intellectual disability. In some individuals, this phenotype ameliorates with age, resulting in a clinical catch-up and normal IQ in adulthood. The post-transcriptional balance of CUX1 expression in the heterozygous brain at late developmental stages appears important for this favorable clinical course.

Notch Signaling Plays a Dual Role in Regulating the Neuron-to-Oligodendrocyte Switch in the Developing Dorsal Forebrain.

Neural progenitor cells in the developing dorsal forebrain generate excitatory neurons followed by oligodendrocytes (OLs) and astrocytes. However, the specific mechanisms that regulate the timing of this neuron-glia switch are not fully understood. In this study, we show that the proper balance of Notch signaling in dorsal forebrain progenitors is required to generate oligodendrocytes during late stages of embryonic development. Using ex vivo and in utero approaches in mouse embryos of both sexes, we found that Notch inhibition reduced the number of oligodendrocyte lineage cells in the dorsal pallium. However, Notch overactivation also prevented oligodendrogenesis and maintained a progenitor state. These results point toward a dual role for Notch signaling in both promoting and inhibiting oligodendrogenesis, which must be fine-tuned to generate oligodendrocyte lineage cells at the right time and in the right numbers. We further identified the canonical Notch downstream factors HES1 and HES5 as negative regulators in this process. CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9-mediated knockdown of Hes1 and Hes5 caused increased expression of the pro-oligodendrocyte factor ASCL1 and led to precocious oligodendrogenesis. Conversely, combining Notch overactivation with ASCL1 overexpression robustly promoted oligodendrogenesis, indicating a separate mechanism of Notch that operates synergistically with ASCL1 to specify an oligodendrocyte fate. We propose a model in which Notch signaling works together with ASCL1 to specify progenitors toward the oligodendrocyte lineage but also maintains a progenitor state through Hes-dependent repression of Ascl1 so that oligodendrocytes are not made too early, thus contributing to the precise timing of the neuron-glia switch.SIGNIFICANCE STATEMENT Neural progenitors make oligodendrocytes after neurogenesis starts to wind down, but the mechanisms that control the timing of this switch are poorly understood. In this study, we identify Notch signaling as a critical pathway that regulates the balance between progenitor maintenance and oligodendrogenesis. Notch signaling is required for the oligodendrocyte fate, but elevated Notch signaling prevents oligodendrogenesis and maintains a progenitor state. We provide evidence that these opposing functions are controlled by different mechanisms. Before the switch, Notch signaling through Hes factors represses oligodendrogenesis. Later, Notch signaling through an unknown mechanism promotes oligodendrogenesis synergistically with the transcription factor ASCL1. Our study underscores the complexity of Notch and reveals its importance in regulating the timing and numbers of oligodendrocyte production.

Electrochemical Assay Validation of Cancer Biomarkers for Clinical Studies

The high mortality rate of ovarian cancer is due to the late stage of detection and development of resistance despite high initial response rates to chemotherapy in about 60-70% of patients. Hence, detailed analytical and clinical assays' validation is crucial for successful translation to clinical cancer treatment, control, and prevention. We are a team of multidisciplinary researchers focusing on an electrochemical biosensor method development and evaluation for clinically significant circulating cancer biomarkers in biofluids. Electrochemical approaches offer simplicity (analogous to the personalized glucometer), shorter assay time than traditional biological assays, high analytical sensitivity combined with biological selectivity, high reproducibility (within ±5–10% STDEV among replicates) and ultra-low detection capabilities offered by the appropriate surface designs (e.g., femtomolar and lower), smaller sample (10 µL) and reagent volumes (50 µL per sample), semi-automation (microfluidics or multichannel sample dispenser), label and label-free options, throughput designs, and direct biofluid analysis (with a moderate 2–5 fold sample dilution) for minimally invasive (e.g., serum) and noninvasive analyses (e.g., saliva, urine, sweat, and tears). This presentation will discuss our pyrenyl-nanocarbon-based electrochemical immunosensor measuring cancer protein concentrations in clinical serum solutions.

Effects of Saline-Alkali Stress on Sugar Metabolism of Jujube Fruit: A Metabolomic Analysis

There have been numerous studies on the effects of salt stress on jujube fruit; however, only a few studies have reported the changes in fruit quality, particularly sugar content, under conditions of combined salt and alkali stress. Therefore, the present study aimed to investigate the performance of jujube fruits under Saline-Alkali stress and the changes in sugar content in fruits. To achieve this, jujube fruits were treated with varying concentrations of salt and alkali during five developmental periods. The content of relevant sugar components was determined, and metabolomics data were analyzed in combination with relevant quantitative gene data. The results indicated that 100 days after flowering, the surface color of jujube fruit gradually turned red, and the cell structure of the fruit gradually loosened with increasing salt and alkali concentration. The content of sugar components at each stage showed that glucose and fructose primarily accumulate in the early stage of development, while sucrose is the main component in the later stage of development. Metabolomic correlation network maps showed that six differentially accumulated metabolites were closely related to specific genes. Among these metabolites, sucrose was identified as the core metabolite in the metabolic pathway. Quantitative analysis of the related genes revealed that ZjvINV2 and ZjHK2 exhibited prolonged adaptability to stress. Additionally, the expression levels of ZjSS1 and ZjSPS2 under Saline-Alkali stress were consistent with the trend of sucrose content during the same period. In conclusion, the variations in sugar content in jujube fruits during different growth stages and under Saline-Alkali treatment conditions were recorded as reference data, and the primary metabolic substances and related regulatory genes produced in jujube fruits under Saline-Alkali stress were preliminarily identified.

Mammal Reproductive Homeobox (Rhox) Genes: An Update of Their Involvement in Reproduction and Development.

The reproductive homeobox on the X chromosome (RHOX) genes were first identified in the mouse during the 1990s and have a crucial role in reproduction. In various transcription factors with a key regulatory role, the homeobox sequence encodes a "homeodomain" DNA-binding motif. In the mouse, there are three clusters of Rhox genes (α, β, and γ) on the X chromosome. Each cluster shows temporal and/or quantitative collinearity, which regulates the progression of the embryonic development process. Although the RHOX family is conserved in mammals, the interspecies differences in the number of RHOX genes and pseudogenes testifies to a rich evolutionary history with several relatively recent events. In the mouse, Rhox genes are mainly expressed in reproductive tissues, and several have a role in the differentiation of primordial germ cells (Rhox1, Rhox6, and Rhox10) and in spermatogenesis (Rhox1, Rhox8, and Rhox13). Despite the lack of detailed data on human RHOX, these genes appear to be involved in the formation of germ cells because they are predominantly expressed during the early (RHOXF1) and late (RHOXF2/F2B) stages of germ cell development. Furthermore, the few variants identified to date are thought to induce or predispose to impaired spermatogenesis and severe oligozoospermia or azoospermia. In the future, research on the pathophysiology of the human RHOX genes is likely to confirm the essential role of this family in the reproductive process and might help us to better understand the various causes of infertility and characterize the associated human phenotypes.

Characterization of some physiological traits of some bread wheat (Triticum aestivum) landraces for drought tolerance at late developmental stages

Characterization of some physiological traits of some bread wheat (Triticum aestivum) landraces for drought tolerance at late developmental stages

Study of Chemical Additives for Optimization of Binary Systems Used for Downhole Thermochemical Treatment of Heavy Oil

Currently, most explored oil fields in Russia are at a late stage of development, and in order to maintain high levels of oil production, it is rational to put into operation fields with hard-to-recover reserves. For complicated oil fields, in particular fields with high-viscosity oil, the known traditional methods of development are ineffective. Therefore, the search for new technologies for the development and operation of such fields to significantly increase oil recovery and intensify production is of fundamental importance. One such method of heat treatment of the bottomhole formation zone is the use of heat and gas generating systems on site. In this work, new results were obtained on physical modeling of thermochemical reaction initiation with delayed-action catalyst (2,2-bis(hydroxymethyl)butanoic acid) filtration tests on composite core models of sandstone and carbonate with foam heat generation and initiating additives of binary type. Using hydrodynamic modelling, the results of laboratory studies were reproduced, and the preliminary efficiency of the developed technology for thermochemical treatment of deposits in the Samara region (Russia) was evaluated.

Identification of watermelon H3K4 and H3K27 genes and their expression profiles during watermelon fruit development.

The ClaH3K4s and ClaH3K27s gene families are subfamilies of the SET family, each with a highly conserved SET structure domain and a PHD structural domain. Both participate in histone protein methylation, which affects the chromosome structure and gene expression, and is essential for fruit growth and development. In order to demonstrate the structure and expression characteristics of ClaH3K4s and ClaH3K27s in watermelon, members of the watermelon H3K4 and H3K27 gene families were identified, and their chromosomal localization, gene structure, and protein structural domains were analyzed. The phylogeny and covariance of the gene families with other species were subsequently determined, and the expression profiles were obtained by performing RNA-Seq and qRT-PCR. The watermelon genome had five H3K4 genes with 3207-8043bp nucleotide sequence lengths and four H3K27 genes with a 1107-5499bp nucleotide sequence. Synteny analysis revealed the close relationship between watermelon and cucumber, with the majority of members displaying a one-to-one covariance. Approximately half of the 'Hua-Jing 13 watermelon' ClaH3K4s and ClaH3K27s genes were expressed more in the late fruit development stages, while the changes were minimal for the remaining half. H3K4-2 expression was observed to be slightly greater on day 21 compared to other periods. Moreover, ClaH3K27-1 and ClaH3K27-2 were hardly expressed throughout the developing period, and ClaH3K27-4 exhibited the highest expression. These results serve as a basis for further functional characterization of the H3K4 and H3K27 genes in the fruit development of watermelon.

The LEA2 gene sub-family: Characterization, evolution, and potential functions in Camellia oleifera seed development and stress response

Camellia oleifera (C. oleifera) is a woody plant that is important economically due to its oil production. Late embryonic abundant (LEA) proteins are hydrophilic and thermostable proteins that accumulate during the late stages of embryo development, serving vital functions in plant development, seed oil accumulation, and stress responses. However, it remains unclear how LEAs regulate seed size, oil accumulation, and stress responses in C. oleifera. In our study, 47 LEA genes were identified across the C. oleifera genome, which were named based on their chromosomal distribution and separated into LEA2 subgroups. Ancient duplication events resulted in 3 tandem and 8 segmental duplications in the C. oleifera genome. Further analysis of genetic structure, motifs, and cis-elements demonstrated that the evolutionary relationships of the LEA gene family in C. oleifera are highly conserved with many of the LEA genes located in the chloroplast. WGCNA was used to identify modules containing co-expressed genes in different seed developmental stages, and the regulatory network revealed that CoLEA6, CoLEA26, CoLEA17, and CoLEA22 were the hub genes, acting alongside the genes involved in plant growth and abiotic stress. The qPCR analysis of 8 C. oleifera LEA genes under simulated drought stress conditions and gradient shade was conducted, and indicated that CoLEAs responded to drought and shade stresses. Overall, CoLEA22 may be critical for responses to abiotic stress and seed development. This study enable valuable insights into the roles of LEA genes in C. oleifera and establishes a foundation for further research on their function in regulating seed development, oil accumulation and stress response.

Off-target pharmacological activity at various kinases: Potential functional and pathological side effects

In drug discovery, during the lead optimization and candidate characterization stages, novel small molecules are frequently evaluated in a battery of in vitro pharmacology assays to identify potential unintended, off-target interactions with various receptors, transporters, ion channels, and enzymes, including kinases. Furthermore, these screening panels may also provide utility at later stages of development to provide a mechanistic understanding of unexpected safety findings. Here, we present a compendium of the most likely functional and pathological outcomes associated with interaction(s) to a panel of 95 kinases based on an extensive curation of the scientific literature. This panel of kinases was designed by AbbVie based on safety-related data extracted from the literature, as well as from over 20 years of institutional knowledge generated from discovery efforts. For each kinase, the scientific literature was reviewed using online databases and the most often reported functional and pathological effects were summarized. This work should serve as a practical guide for small molecule drug discovery scientists and clinical investigators to predict and/or interpret adverse effects related to pharmacological interactions with these kinases.

Molecular simulation study of liquid sulfur adsorption and diffusion in calcite nanoscale slits under varying temperature and pressure conditions

In the late stages of extensive development of high-sulfur carbonate gas reservoirs, pressure decline leads to sulfur enrichment, and when the temperature exceeds sulfur's melting point, sulfur precipitates in a liquid form, severely restricting the exploitation of high-sulfur carbonate gas reservoirs. However, the adsorption mechanism of sulfur deposition remains poorly understood. In this study, we investigated the microscopic adsorption mechanism of liquid sulfur in calcite nanopores using molecular simulation methods. Our research revealed that liquid sulfur adsorption in calcite nanopores is primarily influenced by pore size. When the pore size is less than 160 Å, multilayer adsorption dominates, with a single-layer sulfur film thickness of approximately 5 Å. At the nanoscale, 1 g of calcite with a 40 Å nanopore structure exhibits excess adsorption of liquid sulfur ranging from 0.00734 to 0.0279 g. As pressure increases, the adsorption of two or more layers intensifies, while the diffusion coefficient decreases. Conversely, as temperature rises, the density of each layer decreases, and the diffusion coefficient increases. Simultaneously, the diffusion coefficient of liquid sulfur in calcite slits exhibits a power function increase as the pore size enlarges. Under a pressure of 15 MPa, when the temperature increases from 420 K to 480 K, the diffusion coefficient increases from 6.96 × 10−11 m²/s to 3.51 × 10−10 m²/s. This study provides valuable insights into the adsorption and diffusion behavior of liquid sulfur in calcite nanopores, which can inform the development of high-sulfur carbonate reservoirs.

Expression analysis of alpha keratins and corneous beta-protein genes during embryonic development of Gekko japonicus

Epidermal appendages of birds and reptiles, including claws, feathers, scales, and setae, are primarily composed of alpha keratins (KRTs) and corneous beta-proteins (CBPs). A comprehensive and systematic knowledge of KRTs and CBPs in Schlegel's Japanese gecko (Gekko japonicus) is still lacking. In this study, 22 candidate Gecko japonicus keratin (GjKRT) family genes (12 type I genes, 10 type II genes) were identified in the G. japonicus genome. The majority of GjKRT genes across various subgroups had undergone a prolonged and highly conservative evolutionary process. Through a combination of morphological observation, RNA-seq analysis, and qRT-PCR assay, it was possible to discern the dynamic alterations in the expression of GjKRTs and Gecko japonicus corneous beta-proteins genes (GjCBPs). These findings strongly indicate that GjKRTs gradually accumulate to constitute an α-layer, which is subsequently succeeded by the formation of the corneous beta layer containing GjCBPs at late stages (40–42) of embryonic development. The epidermal appendages in G. japonicus may result from the joint accumulation of KRTs and CBPs, with stages 40–42 being critical for their development. These findings provide novel insights into KRTs and CBPs of G. japonicus and offer a foundation for investigating the functions of GjKRT and GjCBP gene families. Furthermore, this knowledge contributes to unraveling the molecular mechanisms underlying the formation of epidermal appendages in G. japonicus.

Abstract P1104: Mantarray 3D Engineered Muscle Tissue Platform Demonstrates Clinically-relevant Disease Stratification Of An In Vitro Human Duchenne Muscular Dystrophy Model

Accurately modeling disease in vitro is vital for the development of new treatment strategies and therapeutics. For cardiac diseases, direct assessment of contractile output is the “ultimate phenotype” and a reliable metric to study overall tissue function, as other ‘proxy’ measurements or biomarkers are not good predictors of muscle strength. Human 3D engineered heart tissues (EHTs) from induced pluripotent stem cells hold great potential for modeling contractile function. However, the bioengineering strategies required to generate these predictive models present limitations for many investigators. Here, we have developed an instrument platform that utilizes 3D EHTs in conjunction with a label-free, magnetic sensing array (Mantarray). The platform enables facile and reproducible fabrication of 3D EHTs using virtually any cell source and is coupled with parallel direct measurement of contractility. This approach enables clinically relevant functional measurements of muscle, stratification of healthy vs. diseased muscle phenotypes, and facilitates therapeutic modality-agnostic compound safety and efficacy screening. We present a 3D model of Duchenne muscular dystrophy (DMD) that utilizes EHTs formed from an isogenic pair of healthy and diseased cells. DMD tissues display functional deficits across numerous metrics of contractility, including force and relaxation kinetics, though beat rate remains unchanged. EHTs remain functional over months in culture and provide a large experimental window to not only study therapeutic effect, but also disease phenotypes that may present at later stages of development and maturity. We show both acute and chronic effects of compounds in EHTs, including a drug (BMS-986094) that failed clinical trials due to unanticipated cardiotoxicity. These data demonstrate a first-and-only commercial platform that integrates individual, well-based control of electrical stimulation across a 24-well plate of tissues to pace muscle constructs, model force-frequency, and enable exercise regimens or damage protocols. Stimulation is coupled with automated assessment of muscle contraction, providing an inclusive, medium-throughput platform for disease modeling and therapeutic discovery.

The impact of industrial structure adjustment on the spatial industrial linkage of carbon emission: From the perspective of climate change mitigation

Industrial structure adjustment is an effective way to reduce carbon emissions. When adjusting the industrial structure, changes in the input and output of industrial sectors in one region may cause changes in industrial sectors of other regions, resulting in inter-regional industrial linkages. What is the impact of industrial structure adjustment on the linkages, and how does it impact climate change? The modified gravity model combines the influence and inductive coefficients to build the carbon emission industrial spatial linkage network (ISL) using social network analysis (SNA) in this work with the data of China's 2008–2019 provincial carbon emissions. The impact of industrial structure adjustment on carbon emission industrial spatial linkage network and its effect on climate change mitigation are explored with the help of the STIRPAT model and quality assurance procedure (QAP). The results of this study are as follows: (1) The provincial carbon emissions under industrial spatial linkages show significant network characteristics, the network density increased from 0.1437 in 2008 to 0.323 in 2019. (2) There is a significant spatial spillover effect in the industrial spatial linkage network (2019) centered on Shandong and Jiangsu. The provinces can be divided into 4 blocks, which play different roles in the network. (3) The industrial structure upgrading can significantly promote the carbon emission industrial linkage network and mitigate climate change. (4) The industrial structure rationalization can promote the development of the network and mitigate climate change in the early and late development stages of carbon emissions industrial linkage. In addition, it is necessary for the government to pay more attention to the development level of the carbon emission spatial industrial linkage network when implementing industrial structure adjustment in response to climate change.