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Poultry Science Jun 2024The utilization of chicken oviductal epithelial cells (OECs) as a bioreactor to produce therapeutic proteins has shown promise, but the time taken to obtain transgenic...
The utilization of chicken oviductal epithelial cells (OECs) as a bioreactor to produce therapeutic proteins has shown promise, but the time taken to obtain transgenic offspring impedes efficient validation of protein production. To overcome this barrier, we focused on the immortalization of chicken OECs (cOECs) using retroviral vector-mediated c-MYC oncogene expression to establish an in vitro pre-validation system for chicken bioreactors. The resulting immortalized cOECs exhibited sustained proliferation, maintained a normal diploid chicken karyotype, and expressed key oviduct-specific genes (OVA, OVM, LYZ, AVD, and ESR1). Notably, hormonal administration of diethylstilbestrol (DES) or progesterone (P) upregulated oviduct-specific genes in these cells. To enhance the utility of these immortalized cOECs as an in vitro validation system for chicken bioreactors, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology was employed to knock-in (KI) an enhanced green fluorescence protein (EGFP) gene at the ovalbumin (OVA) locus. The resulting OVA EGFP KI immortalized cOECs secreted both EGFP and OVA proteins into the culture medium, with secretion enhanced under DES treatment. This successful integration of an exogenous gene into cOECs enhances their potential as a versatile in vitro validation system for chicken bioreactors. The established immortalized cOECs overcome previous challenges associated with long-term culture and maintenance, providing a reliable platform for efficient protein production validation. This study presents a comprehensive characterization of the immortalized cOECs, addressing critical limitations associated with in vivo systems and laying a foundation for the development of a streamlined and effective chicken bioreactor model.
Topics: Animals; Chickens; Bioreactors; Oviducts; Epithelial Cells; Female; Ovalbumin; Green Fluorescent Proteins
PubMed: 38652946
DOI: 10.1016/j.psj.2024.103723 -
Molecular Biology Reports Apr 2024Phosphodiesterases (PDEs) have become a promising therapeutic target for various disorders. PDEs are a vast and diversified family of enzymes that degrade cyclic... (Review)
Review
Phosphodiesterases (PDEs) have become a promising therapeutic target for various disorders. PDEs are a vast and diversified family of enzymes that degrade cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), which have several biochemical and physiological functions. Phosphodiesterase 4 (PDE4) is the most abundant PDE in the central nervous system (CNS) and is extensively expressed in the mammalian brain, where it catalyzes the hydrolysis of intracellular cAMP. An alteration in the balance of PDE4 and cAMP results in the dysregulation of different biological mechanisms involved in neurodegenerative diseases. By inhibiting PDE4 with drugs, the levels of cAMP inside the cells could be stabilized, which may improve the symptoms of mental and neurological disorders such as memory loss, depression, and Parkinson's disease (PD). Though numerous studies have shown that phosphodiesterase 4 inhibitors (PDE4Is) are beneficial in PD, there are presently no approved PDE4I drugs for PD. This review presents an overview of PDE4Is and their effects on PD, their possible underlying mechanism in the restoration/protection of dopaminergic cell death, which holds promise for developing PDE4Is as a treatment strategy for PD. Methods on how these drugs could be effectively delivered to develop as a promising treatment for PD have been suggested.
Topics: Animals; Humans; Cyclic Nucleotide Phosphodiesterases, Type 4; Parkinson Disease; Phosphodiesterase 4 Inhibitors; Cyclic AMP; Neurodegenerative Diseases; Cyclic GMP; Mammals; Diethylstilbestrol
PubMed: 38622307
DOI: 10.1007/s11033-024-09484-8 -
Communications Biology Apr 2024Erectile dysfunction (ED) is an extremely prevalent condition which significantly impacts quality of life. The rapid increase of ED in recent decades suggests the...
Erectile dysfunction (ED) is an extremely prevalent condition which significantly impacts quality of life. The rapid increase of ED in recent decades suggests the existence of unidentified environmental risk factors contributing to this condition. Endocrine Disrupting Chemicals (EDCs) are one likely candidate, given that development and function of the erectile tissues are hormonally dependent. We use the estrogenic-EDC diethylstilbestrol (DES) to model how widespread estrogenic-EDC exposure may impact erectile function in humans. Here we show that male mice chronically exposed to DES exhibit abnormal contractility of the erectile tissue, indicative of ED. The treatment did not affect systemic testosterone production yet significantly increased estrogen receptor α (Esr1) expression in the primary erectile tissue, suggesting EDCs directly impact erectile function. In response, we isolated the erectile tissue from mice and briefly incubated them with the estrogenic-EDCs DES or genistein (a phytoestrogen). These acute-direct exposures similarly caused a significant reduction in erectile tissue contractility, again indicative of ED. Overall, these findings demonstrate a direct link between estrogenic EDCs and erectile dysfunction and show that both chronic and acute estrogenic exposures are likely risk factors for this condition.
Topics: Humans; Male; Mice; Animals; Endocrine Disruptors; Erectile Dysfunction; Quality of Life; Risk Factors
PubMed: 38565966
DOI: 10.1038/s42003-024-06048-1 -
Cells Mar 2024Repurposing previously approved drugs may fast track the route to the clinic for potential senotherapeutics and improves the inefficiency of the clinical drug...
Repurposing previously approved drugs may fast track the route to the clinic for potential senotherapeutics and improves the inefficiency of the clinical drug development pipeline. We performed a repurposing screen of 240 clinically approved molecules in human primary dermal fibroblasts for their effects on expression. Molecules demonstrating effects on expression underwent secondary screening for senescence-associated beta galactosidase (SAB) activity, based on effect size, direction, and/or molecule identity. Selected molecules then underwent a more detailed assessment of senescence phenotypes including proliferation, apoptosis, DNA damage, senescence-associated secretory phenotype (SASP) expression, and regulators of alternative splicing. A selection of the molecules demonstrating effects on senescence were then used in a new bioinformatic structure-function screen to identify common structural motifs. In total, 90 molecules displayed altered expression at one or other dose, of which 15 also displayed effects on SAB positivity in primary human dermal fibroblasts. Of these, 3 were associated with increased SAB activity, and 11 with reduced activity. The female synthetic sex hormones-diethylstilboestrol, ethynyl estradiol and levonorgestrel-were all associated with a reduction in aspects of the senescence phenotype in male cells, with no effects visible in female cells. Finally, we identified that the 30 compounds that decreased activity the most had a common substructure linked to this function. Our results suggest that several drugs licensed for other indications may warrant exploration as future senotherapies, but that different donors and potentially different sexes may respond differently to senotherapeutic compounds. This underlines the importance of considering donor-related characteristics when designing drug screening platforms.
Topics: Male; Humans; Female; Cellular Senescence; Senotherapeutics; Drug Repositioning; Hormones
PubMed: 38534362
DOI: 10.3390/cells13060517 -
Nanotechnology Mar 2024In this research, the MIL-47/ACET/Nafion/GCE electrochemical senser for the determination of diethylstilbestrol (DES) was prepared with vanadyl sulfate (VOSO·HO) and...
In this research, the MIL-47/ACET/Nafion/GCE electrochemical senser for the determination of diethylstilbestrol (DES) was prepared with vanadyl sulfate (VOSO·HO) and terephthalic acid (HBDC) as the main raw materials, compounded with acetylene black (ACET) and perfluorosulfonic acid polymer (Nafion). The compound DES belongs to the category of estrogens, and prolonged exposure to the environment can have detrimental effects on the physiological functioning of both humans and animals. Due to the strong DES enrichment performance of MIL-47(V-MOFs) with large specific surface area, in addition to the excellent conductivity and electrocatalysis of composite materials, this modified senser had good electrochemical response to DES. With differential pulse voltammetry, in optimum condition of 0.1 M NaHPO-NaHPOat pH = 7.0, potential interval of -1.0 to 1.0 V, enrichment time of 120 s and enrichment potential of 0.2 V, there was a good linear relationship between peak current and the concentration of DES over the range of 0.1 and 50M, and the limit of detection was 0.008M. The sensor accurately detected DES in actual water samples, with recovery rates ranging from 89.21% to 105.3%. The electrochemical sensor was simple to prepare and had practical significance for the detection of DES in water. The research results of the sensor provide another alternative analytical means for the sensitive detection of DES in the environment, which is important for maintaining public health.
PubMed: 38529942
DOI: 10.1088/1361-6528/ad321d -
Frontiers in Immunology 2024T-cell activation is a pivotal process of the adaptive immune response with 3',5'-cyclic adenosine monophosphate (cAMP) as a key regulator of T-cell activation and... (Review)
Review
T-cell activation is a pivotal process of the adaptive immune response with 3',5'-cyclic adenosine monophosphate (cAMP) as a key regulator of T-cell activation and function. It governs crucial control over T-cell differentiation and production of pro-inflammatory cytokines, such as IFN-γ. Intriguingly, levels of intracellular cAMP differ between regulatory (Treg) and conventional T-cells (Tcon). During cell-cell contact, cAMP is transferred via gap junctions between these T-cell subsets to mediate the immunosuppressive function of Treg. Moreover, the activation of T-cells via CD3 and CD28 co-stimulation leads to a transient upregulation of cAMP. Elevated intracellular cAMP levels are balanced precisely by phosphodiesterases (PDEs), a family of enzymes that hydrolyze cyclic nucleotides. Various PDEs play distinct roles in regulating cAMP and cyclic guanosine monophosphate (cGMP) in T-cells. Research on PDEs has gained growing interest due to their therapeutic potential to manipulate T-cell responses. So far, PDE4 is the best-described PDE in T-cells and the first PDE that is currently targeted in clinical practice to treat autoimmune diseases. But also, other PDE families harbor additional therapeutic potential. PDE2A is a dual-substrate phosphodiesterase which is selectively upregulated in Tcon upon activation. In this Mini-Review, we will highlight the impact of cAMP regulation on T-cell activation and function and summarize recent findings on different PDEs regulating intracellular cAMP levels in T-cells.
Topics: Phosphoric Diester Hydrolases; Phosphodiesterase Inhibitors; Cyclic AMP; T-Lymphocytes; Diethylstilbestrol
PubMed: 38524120
DOI: 10.3389/fimmu.2024.1365484 -
Neural Networks : the Official Journal... Jun 2024In this paper, we introduce PDE-LEARN, a novel deep learning algorithm that can identify governing partial differential equations (PDEs) directly from noisy, limited...
In this paper, we introduce PDE-LEARN, a novel deep learning algorithm that can identify governing partial differential equations (PDEs) directly from noisy, limited measurements of a physical system of interest. PDE-LEARN uses a Rational Neural Network, U, to approximate the system response function and a sparse, trainable vector, ξ, to characterize the hidden PDE that the system response function satisfies. Our approach couples the training of U and ξ using a loss function that (1) makes U approximate the system response function, (2) encapsulates the fact that U satisfies a hidden PDE that ξ characterizes, and (3) promotes sparsity in ξ using ideas from iteratively reweighted least-squares. Further, PDE-LEARN can simultaneously learn from several data sets, allowing it to incorporate results from multiple experiments. This approach yields a robust algorithm to discover PDEs directly from realistic scientific data. We demonstrate the efficacy of PDE-LEARN by identifying several PDEs from noisy and limited measurements.
Topics: Deep Learning; Algorithms; Diethylstilbestrol; Neural Networks, Computer
PubMed: 38521016
DOI: 10.1016/j.neunet.2024.106242 -
Chemosphere May 2024Growing focus has been drawn to the continuous detection of high estrogens levels in the soil environment. Additionally, microplastics (MPs) are also of growing concern...
Growing focus has been drawn to the continuous detection of high estrogens levels in the soil environment. Additionally, microplastics (MPs) are also of growing concern worldwide, which may affect the environmental behavior of estrogens. However, little is known about effects of MPs occurrence on estrogens degradation in soil. In this study, polyethylene microplastics (PE-MPs) were chosen to examine the influence on six common estrogens (estrone (E1), 17α-estradiol (17α-E2), 17β-estradiol (17β-E2), estriol (E3), diethylstilbestrol (DES), and 17α-ethinylestradiol (17α-EE2)) degradation. The results indicated that PE-MPs had little effect on the degradation of E3 and DES, and slightly affected the degradation of 17α-E2, however, significantly inhibited the degradation of E1, 17α-EE2, and 17β-E2. It was explained that (i) obvious oxidation reaction occurred on the surface of PE-MPs, indicating that PE-MPs might compete with estrogens for oxidation sites, such as redox and biological oxidation; (ii) PE-MPs significantly changed the bacterial community in soil, resulting in a decline in the abundance of some bacterial communities that biodegraded estrogens. Moreover, the rough surface of PE-MPs facilitated the estrogen-degrading bacterial species (especially for E1, E2, and EE2) to adhere, which decreased their reaction to estrogens. These findings are expected to deepen the understanding of the environmental behavior of typical estrogens in the coexisting system of MPs.
Topics: Microplastics; Plastics; Polyethylene; Soil; Estradiol; Estrogens; Estrone; Ethinyl Estradiol
PubMed: 38499076
DOI: 10.1016/j.chemosphere.2024.141727