Differential Modulation Research Articles

Identification of shared gene signatures and pathways for diagnosing osteoporosis with sarcopenia through integrated bioinformatics analysis and machine learning

BackgroundPrior studies have suggested a potential relationship between osteoporosis and sarcopenia, both of which can present symptoms of compromised mobility. Additionally, fractures among the elderly are often considered a common outcome of both conditions. There is a strong correlation between fractures in the elderly population, decreased muscle mass, weakened muscle strength, heightened risk of falls, and diminished bone density. This study aimed to pinpoint crucial diagnostic candidate genes for osteoporosis patients with concomitant sarcopenia.MethodsTwo osteoporosis datasets and one sarcopenia dataset were obtained from the Gene Expression Omnibus (GEO). Differential expression genes (DEGs) and module genes were identified using Limma and Weighted Gene Co-expression Network Analysis (WGCNA), followed by functional enrichment analysis, construction of protein–protein interaction (PPI) networks, and application of a machine learning algorithm (least absolute shrinkage and selection operator (LASSO) regression) to determine candidate hub genes for diagnosing osteoporosis combined with sarcopenia. Receiver operating characteristic (ROC) curves and column line plots were generated.ResultsThe merged osteoporosis dataset comprised 2067 DEGs, with 424 module genes filtered in sarcopenia. The intersection of DEGs between osteoporosis and sarcopenia module genes consisted of 60 genes, primarily enriched in viral infection. Through construction of the PPI network, 30 node genes were filtered, and after machine learning, 7 candidate hub genes were selected for column line plot construction and diagnostic value assessment. Both the column line plots and all 7 candidate hub genes exhibited high diagnostic value (area under the curve ranging from 1.00 to 0.93).ConclusionWe identified 7 candidate hub genes (PDP1, ALS2CL, VLDLR, PLEKHA6, PPP1CB, MOSPD2, METTL9) and constructed column line plots for osteoporosis combined with sarcopenia. This study provides reference for potential peripheral blood diagnostic candidate genes for sarcopenia in osteoporosis patients.

Lipid metabolism: a central modulator of RORγt-mediated Th17 cell differentiation.

Among the T helper cell subsets, Th17 cells contribute to the development of various inflammatory and autoimmune diseases, including psoriasis, rheumatoid arthritis, inflammatory bowel disease, steroid-resistant asthma, and multiple sclerosis. Retinoid-related orphan receptor gamma t (RORγt), a nuclear hormone receptor, serves as a master transcription factor for Th17 cell differentiation. Recent findings have shown that modulating the metabolic pathway is critical for Th17 cell differentiation, particularly through the engagement of de novo lipid biosynthesis. Suppression of lipid biosynthesis, either through the pharmacological inhibition or gene deletion of related enzymes in CD4+ T cells, results in significant impairment of Th17 cell differentiation. Mechanistic studies indicate that metabolic fluxes through both the fatty acid and cholesterol biosynthetic pathways have a pivotal role in the regulation of RORγt activity through the generation of endogenous RORγt lipid ligands. This review discusses recent discoveries highlighting the importance of lipid metabolism in Th17 cell differentiation and function, as well as exploring specific molecular pathways involved in RORγt activation through cellular lipid metabolism. We further elaborate on a pioneering therapeutic approach to improve inflammatory and autoimmune disorders via the inhibition of RORγt.

A new application of Cu2+ on differential modification to promote copper-molybdenum separation with a novel chalcopyrite depressant amidinothiourea

The separation of chalcopyrite and molybdenite has always been a challenge in mineral processing because of their similar floatability and close association. This study proposes a new chalcopyrite depressant named amidinothiourea (ADT) and a new application of Cu2+ on differentially modifying chalcopyrite and molybdenite. It is found that small dosage of ADT is insufficient to depress chalcopyrite, although there is ADT adsorption on chalcopyrite surface at this time. While after Cu2+ modification on chalcopyrite and molybdenite, a small dosage of ADT can strongly depress chalcopyrite, and has no effect on molybdenite. The largest difference of grade and recovery between chalcopyrite and molybdenite were 45.32 and 70.79 % in mixed-mineral flotation with 2.0 × 10−4 mol/L of Cu2+ and 1.6 × 10−4 mol/L of ADT. The mechanisms of ADT depression and Cu2+ modification were investigated through LEIS, XPS, ToF-SIMS and ICP-OES tests. LEIS experiments show that ADT can adsorb on chalcopyrite surface, and particularly pronounced after the modification of Cu2+ on chalcopyrite. XPS and ToF-SIMS confirm that N is the site of action for ADT, and Cu is the site of action for minerals. XPS, ToF-SIMS and ICP-OES tests reveal that Cu2+ modifies chalcopyrite by promoting the dissolution of iron ions and firmly binding with the exposed unsaturated S. While Cu2+ modifies molybdenite by binding with saturated S atoms weakly.

Miura Origami‐Inspired Reconfigurable Phase Gradient Metasurface for Linearly and Circularly Polarized Differential Modulation

Miura origami's reconfigurable characteristic and structural asymmetry, combined with electromagnetic (EM) wave manipulation, crashed a unique spark. However, the complexity of the three‐dimensional (3D) origami structure after folding makes it challenging to study the phase regulation mechanism. Here, we propose a reconfigurable phase gradient metasurface based on Miura origami and derive the underlying mechanism of phase modulation in detail under linearly and circularly polarized (LP and CP) incidence. We adopt the one‐dimensional (1D) gradient design along the x direction to verify the idea. The phase calculation formulas are given under LP and CP incidence through the Jones matrix's derivation. The beam deflector angles corresponding to LP and CP waves are identical in the planar state. As the folding angle increases, the phase evolution rules corresponding to the LP and CP waves are discrepant, leading to differential beam steering. Finally, the origami sample is processed for verification, and the experimental data are consistent with the simulation and theoretically calculated values. We believe this work can help analyze the EM behavior of complex 3D origami structures and lay a foundation for designing a multifunctional EM origami metasurface.

Exploring Vitamin D Synthesis and Function in Cardiovascular Health: A Narrative Review

Vitamin D plays a crucial role in the human body, influencing a wide range of physiological processes from bone health to immune function. The complex biochemical pathways involved in the synthesis, metabolism, and action of Vitamin D are explored, emphasizing its importance in nutrition and food technology. This review also investigates the regulatory mechanisms that control Vitamin D metabolism and its systemic effects on calcium homeostasis, cell proliferation, differentiation, and immune modulation. The role of Vitamin D3 in regulating blood pressure and atherosclerosis in the onset of cardiovascular disorders is discussed. Given the importance of Vitamin D in food science and technology, the regulatory mechanisms that control Vitamin D metabolism and its systemic effects on calcium homeostasis are also investigated, integrating innovative approaches and advanced technologies to improve human health through nutrition. Additionally, the review assesses the influence of food processing on Vitamin D levels and discusses cutting-edge technologies as innovative strategies to mitigate Vitamin D loss during food processing. This comprehensive exploration aims to improve our understanding of the biochemical pathways of Vitamin D and its relevance to food science, contributing to the development of new strategies for food fortification and the promotion of optimal health through diet.

Investigation of Bandgap Properties of a Piezoelectric Phononic Crystal Plate Based on the PDE Module in COMSOL.

Aiming to address the vibration noise problems on ships, we constructed a piezoelectric phononic crystal (PC) plate structure model, solved the governing equations of the structure using the partial differential equations module (PDE) in the finite element softwareCOMSOL6.1, and obtained the corresponding energy band structure, transmission curves, and vibration modal diagrams. The application of this method to probe the structural properties of two-dimensional piezoelectric PCs is described in detail. The calculation results obtained using this method were compared with the structures obtained using the traditional plane wave expansion method (PWE) and the finite element method (FE). The results were found to be in perfect agreement, which verified the feasibility of this method. To safely and effectively adjust the bandgap within a reasonable voltage range, this paper explored the order of magnitude of the plate thickness, the influence of the voltage on the bandgap, and the dependence between them. It was found that the smaller the order of magnitude of the plate thickness, the smaller the order of magnitude of the band in which the bandgap was located. The magnitude of the driving voltage that made the bandgap change became smaller accordingly. The new idea of attaching the PC plate to the conventional plate structure to achieve a vibration damping effect is also briefly introduced. Finally, the effects of lattice constant, plate width, and thickness on the bandgap were investigated.

Stryphnodendron adstringens have a modulatory effect on inflammatory cytokines markers of in vitro activated macrophages

Aim: The purpose of this study is to conduct a comprehensive investigation into the modulatory effects of Stryphnodendron adstringens (Mart.; S. adstringens), a Brazilian wound-healing plant, on the expression of inflammatory cytokines. This will be achieved using an in vitro protocol with the commercial macrophage cell line RAW 264.7. Methods: The macrophage inflammatory response was induced by the natural antigen phytohemagglutinin (PHA), with and without supplementation of different concentrations of S. adstringens extract. The effects on cell proliferation rate and the concentration and production of transcripts of pro-inflammatory cytokines interleukin 1β (IL-1β), IL-6, tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ), as well as the anti-inflammatory cytokine IL-10, were assessed using spectrophotometric, immunoassay, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) techniques. Results: S. adstringens extract at all concentrations tested here reduced the cellular proliferation rate of activated macrophages. Extracts at concentrations of 0.49 mg/mL and 0.99 mg/mL decreased the protein and gene expression of pro-inflammatory cytokines, exhibiting the opposite effect concerning IL-10. Conclusions: The findings suggest that the wound-healing action of S. adstringens may encompass differential modulation of inflammation associated with tissue injury.

Differential Modulation of Catecholamine and Adipokine Secretion by the Short Chain Fatty Acid Receptor FFAR3 and α2-Adrenergic Receptors in PC12 Cells.

Sympathetic nervous system (SNS) hyperactivity is mediated by elevated catecholamine (CA) secretion from the adrenal medulla, as well as enhanced norepinephrine (NE) release from peripheral sympathetic nerve terminals. Adrenal CA production from chromaffin cells is tightly regulated by sympatho-inhibitory α2-adrenergic (auto)receptors (ARs), which inhibit both epinephrine (Epi) and NE secretion via coupling to Gi/o proteins. α2-AR function is, in turn, regulated by G protein-coupled receptor (GPCR)-kinases (GRKs), especially GRK2, which phosphorylate and desensitize them, i.e., uncouple them from G proteins. On the other hand, the short-chain free fatty acid (SCFA) receptor (FFAR)-3, also known as GPR41, promotes NE release from sympathetic neurons via the Gi/o-derived free Gβγ-activated phospholipase C (PLC)-β/Ca2+ signaling pathway. However, whether it exerts a similar effect in adrenal chromaffin cells is not known at present. In the present study, we examined the interplay of the sympatho-inhibitory α2A-AR and the sympatho-stimulatory FFAR3 in the regulation of CA secretion from rat adrenal chromaffin (pheochromocytoma) PC12 cells. We show that FFAR3 promotes CA secretion, similarly to what GRK2-dependent α2A-AR desensitization does. In addition, FFAR3 activation enhances the effect of the physiologic stimulus (acetylcholine) on CA secretion. Importantly, GRK2 blockade to restore α2A-AR function or the ketone body beta-hydroxybutyrate (BHB or 3-hydroxybutyrate), via FFAR3 antagonism, partially suppress CA production, when applied individually. When combined, however, CA secretion from PC12 cells is profoundly suppressed. Finally, propionate-activated FFAR3 induces leptin and adiponectin secretion from PC12 cells, two important adipokines known to be involved in tissue inflammation, and this effect of FFAR3 is fully blocked by the ketone BHB. In conclusion, SCFAs can promote CA and adipokine secretion from adrenal chromaffin cells via FFAR3 activation, but the metabolite/ketone body BHB can effectively inhibit this action.

Role of N6-methyladenosine-related lncRnas in pseudoexfoliation glaucoma

ABSTRACT To explore the role of lncRNA m6A methylation modification in aqueous humour (AH) of patients with pseudoexfoliation glaucoma (PXG). Patients with open-angle PXG under surgery from June 2021 to December 2021 were selected. Age- and gender-matched patients with age-related cataract (ARC) were chosen as control. Patients underwent detailed ophthalmic examinations. 0.05–0.1 ml AH were extracted during surgery for MeRIP-Seq and RNA-Seq. Joint analysis was used to screen lncRNAs with differential m6A methylation modification and expression. Online software tools were used to draw lncRNA-miRNA-mRNA network (ceRNA). Expression of lncRNAs and mRNAs was confirmed using quantitative real-time PCR. A total of 4151 lncRNAs and 4386 associated m6A methylation modified peaks were identified in the PXG group. Similarly, 2490 lncRNAs and 2595 associated m6A methylation modified peaks were detected in the control. Compared to the ARC group, the PXG group had 234 hypermethylated and 402 hypomethylated m6A peaks, with statistically significant differences (| Fold Change (FC) |≥2, p < 0.05). Bioinformatic analysis revealed that these differentially methylated lncRNA enriched in extracellular matrix formation, tight adhesion, TGF- β signalling pathway, AMPK signalling pathway, and MAPK signalling pathway. Joint analysis identified 10 lncRNAs with differential m6A methylation and expression simultaneously. Among them, the expression of ENST000000485383 and ROCK1 were confirmed downregulated in the PXG group by RT-qPCR. m6A methylation modification may affect the expression of lncRNA and participate in the pathogenesis of PXG through the ceRNA network. ENST000000485383-hsa miR592-ROCK1 May be a potential target pathway for further investigation in PXG m6A methylation.

Cipher-image compression by using compressive sensing and interweaving permutation

Recently, compressing cipher images by using compressive sensing (CS) has attracted a lot of attention. However, the previous CS-based works cannot provide satisfactory compression performance. In this article, a cipher-image compression scheme based on CS and interweaving permutation is proposed to address this concern. Before image compression, the nature image is down-sampled to generate a lot of patches by using interweaving permutation firstly and then each patch is encrypted by a scrambling matrix, where each patch is a down-sampled image. In this way, the encrypted patches are highly correlated from one to each other since they preserve the correlation of the original patches. Therefore, we can encode the cipher image by employing CS and differential pulse-code modulation (DPCM) in an efficient way. To reconstruct the image effectively, an iterative shrinkage-thresholding (IST) algorithm is developed. Compared with the previous CS-based works, our scheme achieves better ratio-distortion (R-D) performance.

GnRH-driven FTO-mediated RNA m6A modification promotes gonadotropin synthesis and secretion

BackgroundGonadotropin precisely controls mammalian reproductive activities. Systematic analysis of the mechanisms by which epigenetic modifications regulate the synthesis and secretion of gonadotropin can be useful for more precise regulation of the animal reproductive process. Previous studies have identified many differential m6A modifications in the GnRH-treated adenohypophysis. However, the molecular mechanism by which m6A modification regulates gonadotropin synthesis and secretion remains unclear.ResultsHerein, it was found that GnRH can promote gonadotropin synthesis and secretion by promoting the expression of FTO. Highly expressed FTO binds to Foxp2 mRNA in the nucleus, exerting a demethylation function and reducing m6A modification. After Foxp2 mRNA exits the nucleus, the lack of m6A modification prevents YTHDF3 from binding to it, resulting in increased stability and upregulation of Foxp2 mRNA expression, which activates the cAMP/PKA signaling pathway to promote gonadotropin synthesis and secretion.ConclusionsOverall, the study reveals the molecular mechanism of GnRH regulating the gonadotropin synthesis and secretion through FTO-mediated m6A modification. The results of this study allow systematic interpretation of the regulatory mechanism of gonadotropin synthesis and secretion in the pituitary at the epigenetic level and provide a theoretical basis for the application of reproductive hormones in the regulation of animal artificial reproduction.

Abstract PO1-23-11: Differential m6A modification identified by direct RNA sequencing in endocrine- resistant and sensitive breast cancer cells

Abstract A major limitation of current adjuvant treatments for the ~ 70% of breast cancer patients with estrogen receptor α–expressing (ER+) primary breast tumors is the development of acquired and adaptive endocrine therapy (ET) resistance and metastatic spread. Chemical modifications of transcribed RNAs alters their cellular location, stability, and, for mRNAs, translation into proteins. N-6 methyladenosine (m6A) is the most common modification of mRNA. We tested the hypothesis that m6A epitranscriptomic changes alters the transcriptome resulting in changes in targets in pathways contributing to the metastatic cascade in ET-resistant breast cancer cells. We used an unbiased approach of direct mRNA sequencing using the Oxford Nanopore MinION platform on polyA-selected mRNA from MCF-7 luminal A breast cancer cells and ER+, ET-resistant LCC9 breast cancer cells treated with vehicle control or 100 nM 4-hydroxytamoxifen (4-OHT) for 24 h. In addition, MCF-7 cells were treated with the METTL3 (the catalytic methyltransferase that adds the methyl group from S-adenosylmethionine (SAM) to the adenosine on the N6 position) inhibitor STM2457 to identify m6A positions and transcripts directly regulated by METTL3 activity. Computational analysis directly identified m6A locations in transcripts. We examined the differential modification rate (DMR) between MCF-7 cells +/STM2457 treatment, between MCF-7 and LCC9 cells with vehicle control treatment or with 4-OHT treatment and within each cell line with 4-OHT treatment. We identified 183 and 3,097 genes with m6A DMRs between vehicle-treated and 4-OHT-treated MCF-7 and LCC9 cells, respectively. For 4-OHT-treated cells, we identified 2,401 and 2,638 genes with m6A DMRs in MCF-7 and LCC9 cells respectively with 1,760 genes in common. We identified many genes with multiple m6A modifications, e.g., AMIGO2 appeared to have 14 m6A sites at DRACH motifs in MCF-7 cells, but none were m6A modified in LCC9 cells. Enrichment by Pathway Maps, Process Networks, and GO Processes were performed. Overall, our data suggest a role for altered m6A positions in transcripts and pathways involved in ET-resistance in breast cancer. Citation Format: Belinda Petri, Bailey Valdes, Kellianne Piell, Eric Rouchka, Carolyn Klinge. Differential m6A modification identified by direct RNA sequencing in endocrine- resistant and sensitive breast cancer cells [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-23-11.

Abstract 3089: Do Arteries And Veins Have Different Remodeling Programs?

Aims: Arteries and veins respond differently to injury, with significantly higher rates of postsurgical failure in the latter. The biological reasons for this discrepancy are not well understood, which is a limiting factor for the development of vein-targeted therapies. This study contrasts peripheral arteries and veins at the single-cell level, with a focus on cell populations with remodeling potential. Methods and Results: Upper arm arteries (brachial) and veins (basilic/cephalic) from 27 organ donors were compared using a combination of bulk and single-cell RNA sequencing, proteomics, flow cytometry, and histology. The cellular atlases of six arteries and veins demonstrated a significantly higher proportion of smooth muscle cells (SMCs) in arteries vs. higher counts of fibroblasts in veins. In both types of vessels, clustering analyses identified an intermediate myofibroblast population with repair characteristics. Both arterial and venous myofibroblasts were characterized by upregulation of F2R , FN1, POSTN, COL8A1, and ITGA10 , and were confirmed by histology and F2R/CD90 double staining by flow cytometry. However, bioinformatic analyses indicate that arterial myofibroblasts are phenotypically related to both SMCs and fibroblasts, whereas in veins they are predominantly fibroblast derived. Differences in wall structure further explain the distinct contributions of SMC and fibroblasts to these reparative populations. PDGFRA + fibroblasts are almost exclusively located outside the external elastic lamina in arteries, while widely distributed throughout the venous wall. In line with a higher abundance of fibroblasts and fibroblast-derived reparative cells, ECM-targeted proteomics confirmed a higher abundance of fibrillar collagens in veins vs. more basement ECM components in arteries. These differences in ECM composition are also associated with vessel-specific modulation of myofibroblast differentiation. Conclusions: The distinct cellular origins and transcriptional programs of myofibroblast populations in arteries and veins may explain differences in acute and chronic wall remodeling between vessels. This information is relevant for the development of anti-stenotic therapies.

Research on Frequency Discrimination Method Using Multiplicative-Integral and Linear Transformation Network

In this paper, a frequency discrimination method using a multiplicative-integral and linear transformation network is proposed. In this method, two preset differential frequency signals and frequency modulation signals are transformed by multiplication and integration, and then the instantaneous frequency parameters of the frequency modulation signal are accurately analyzed by the linear transformation network to restore the original modulation signal. Compared with the phase discriminator, the simulation results show that this method has a higher frequency discrimination bandwidth. In addition, this method has better anti-noise performance, and the frequency discrimination distortion caused by noise with a different Signal-to-Noise Ratio is reduced by 33.80% on average compared with the phase discriminator. What is more, the carrier center frequency error has little influence on the frequency discrimination quality of this method, which solves the problem that most common frequency discriminators are seriously affected by the carrier center frequency error. This method requires a low accuracy of carrier center frequency, which makes it extremely suitable for digital frequency discrimination technology and can meet the needs of various frequency discrimination occasions.

Ratiometric Sensing of Azithromycin and Sulfide Using Dual Emissive Carbon Dots: A Turn On-Off-On Approach.

A novel ratiometric fluorescence probe was developed for the determination of azithromycin (AZM) and sulfide ions based on the differential modulation of red emissive carbon dots (R-N@CDs) and blue emissive carbon dots (B-NS@CDs). The addition of sulfide anion selectively quenched the red emission of R-N@CDs while the blue emission of B-NS@CDs unaffected. Upon subsequent introduction of AZM to this R-N@CDs@sulfide system, the quenched red fluorescence was restored. Comprehensive characterization of the CDs was performed using UV-Vis, fluorescence, FTIR spectroscopy, XPS, and TEM. The proposed method exhibited excellent sensitivity and selectivity, with limits of detection of 0.33 µM for AZM and 0.21 µM for sulfide. Notably, this approach enabled direct detection of sulfide without requiring prior modulation of the CDs with metal ions, as is common in other reported methods. The ratiometric probe was successfully applied for the determination of AZM in biological fluids and sulfide in environmental water samples with high selectivity. This work presents the first fluorometric method for the detection of AZM in biological fluids.

Genomic and non-genomic action of vitamin D on ion channels – Targeting mitochondria

Recent studies revealed that mitochondria are not only a place of vitamin D3 metabolism but also direct or indirect targets of its activities. This review summarizes current knowledge on the regulation of ion channels from plasma and mitochondrial membranes by the active form of vitamin D3 (1,25(OH)2D3). 1,25(OH)2D3, is a naturally occurring hormone with pleiotropic activities; implicated in the modulation of cell differentiation, and proliferation and in the prevention of various diseases, including cancer. Many experimental data indicate that 1,25(OH)2D3 deficiency induces ionic remodeling and 1,25(OH)2D3 regulates the activity of multiple ion channels. There are two main theories on how 1,25(OH)2D3 can modify the function of ion channels. First, describes the involvement of genomic pathways of response to 1,25(OH)2D3 in the regulation of the expression of the genes encoding channels, their auxiliary subunits, or additional regulators. Interestingly, intracellular ion channels, like mitochondrial, are encoded by the same genes as plasma membrane channels. Therefore, the comprehensive genomic regulation of the channels from these two different cellular compartments we analyzed using a bioinformatic approach. The second theory explores non-genomic pathways of vitamin D3 activities. It was shown, that 1,25(OH)2D3 indirectly regulates enzymes that impact ion channels, change membrane physical properties, or directly bind to channel proteins. In this article, the involvement of genomic and non-genomic pathways regulated by 1,25(OH)2D3 in the modulation of the levels and activity of plasma membrane and mitochondrial ion channels was investigated by an extensive review of the literature and analysis of the transcriptomic data using bioinformatics.

Thapsigargin and Tunicamycin Block SARS-CoV-2 Entry into Host Cells via Differential Modulation of Unfolded Protein Response (UPR), AKT Signaling, and Apoptosis.

SARS-Co-V2 infection can induce ER stress-associated activation of unfolded protein response (UPR) in host cells, which may contribute to the pathogenesis of COVID-19. To understand the complex interplay between SARS-Co-V2 infection and UPR signaling, we examined the effects of acute pre-existing ER stress on SARS-Co-V2 infectivity. Huh-7 cells were treated with Tunicamycin (TUN) and Thapsigargin (THA) prior to SARS-CoV-2pp transduction (48 h p.i.) to induce ER stress. Pseudo-typed particles (SARS-CoV-2pp) entry into host cells was measured by Bright GloTM luciferase assay. Cell viability was assessed by cell titer Glo® luminescent assay. The mRNA and protein expression was evaluated by RT-qPCR and Western Blot. TUN (5 µg/mL) and THA (1 µM) efficiently inhibited the entry of SARS-CoV-2pp into host cells without any cytotoxic effect. TUN and THA's attenuation of virus entry was associated with differential modulation of ACE2 expression. Both TUN and THA significantly reduced the expression of stress-inducible ER chaperone GRP78/BiP in transduced cells. In contrast, the IRE1-XBP1s and PERK-eIF2α-ATF4-CHOP signaling pathways were downregulated with THA treatment, but not TUN in transduced cells. Insulin-mediated glucose uptake and phosphorylation of Ser307 IRS-1 and downstream p-AKT were enhanced with THA in transduced cells. Furthermore, TUN and THA differentially affected lipid metabolism and apoptotic signaling pathways. These findings suggest that short-term pre-existing ER stress prior to virus infection induces a specific UPR response in host cells capable of counteracting stress-inducible elements signaling, thereby depriving SARS-Co-V2 of essential components for entry and replication. Pharmacological manipulation of ER stress in host cells might provide new therapeutic strategies to alleviate SARS-CoV-2 infection.

Differential Modulation by Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) of Mesenteric Fat and Macrophages and T Cells in Adipose Tissue of Obese fa/fa Zucker Rats.

Polyunsaturated fatty acids (PUFAs) can alter adipose tissue function; however, the relative effects of plant and marine n3-PUFAs are less clear. Our objective was to directly compare the n3-PUFAs, plant-based α-linolenic acid (ALA) in flaxseed oil, and marine-based eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) in high-purity oils versus n6-PUFA containing linoleic acid (LA) for their effects on the adipose tissue and oral glucose tolerance of obese rats. Male fa/fa Zucker rats were assigned to faALA, faEPA, faDHA, and faLA groups and compared to baseline fa/fa rats (faBASE) and lean Zucker rats (lnLA). After 8 weeks, faEPA and faDHA had 11-14% lower body weight than faLA. The oral glucose tolerance and total body fat were unchanged, but faEPA had less mesenteric fat. faEPA and faDHA had fewer large adipocytes compared to faLA and faALA. EPA reduced macrophages in the adipose tissue of fa/fa rats compared to ALA and DHA, while faLA had the greatest macrophage infiltration. DHA decreased (~10-fold) T-cell infiltration compared to faBASE and faEPA, whereas faALA and faLA had an ~40% increase. The n3-PUFA diets attenuated tumour necrosis factor-α in adipose tissue compared to faBASE, while it was increased by LA in both genotypes. In conclusion, EPA and DHA target different aspects of inflammation in adipose tissue.

Cryptosporidium life cycle small molecule probing implicates translational repression and an Apetala 2 transcription factor in macrogamont differentiation.

The apicomplexan parasite Cryptosporidium is a leading cause of childhood diarrhea in developing countries. Current treatment options are inadequate and multiple preclinical compounds are being actively pursued as potential drugs for cryptosporidiosis. Unlike most apicomplexans, Cryptosporidium spp. sequentially replicate asexually and then sexually within a single host to complete their lifecycles. Anti-cryptosporidial compounds are generally identified or tested through in vitro phenotypic assays that only assess the asexual stages. Therefore, compounds that specifically target the sexual stages remain unexplored. In this study, we leveraged the ReFRAME drug repurposing library against a newly devised multi-readout imaging assay to identify small-molecule compounds that modulate macrogamont differentiation and maturation. RNA-seq studies confirmed selective modulation of macrogamont differentiation for 10 identified compounds (9 inhibitors and 1 accelerator). The collective transcriptomic profiles of these compounds indicates that translational repression accompanies Cryptosporidium sexual differentiation, which we validated experimentally. Additionally, cross comparison of the RNA-seq data with promoter sequence analysis for stage-specific genes converged on a key role for an Apetala 2 (AP2) transcription factor (cgd2_3490) in differentiation into macrogamonts. Finally, drug annotation for the ReFRAME hits indicates that an elevated supply of energy equivalence in the host cell is critical for macrogamont formation.

Multiple cholinergic receptor subtypes coordinate dual modulation of acetylcholine on anterior and posterior paraventricular thalamic neurons.

Paraventricular thalamus (PVT) plays important roles in the regulation of emotion and motivation through connecting many brain structures including the midbrain and the limbic system. Although acetylcholine (ACh) neurons of the midbrain were reported to send projections to PVT, little is known about how cholinergic signaling regulates PVT neurons. Here, we used both RNAscope and slice patch-clamp recordings to characterize cholinergic receptor expression and ACh modulation of PVT neurons in mice. We found ACh excited a majority of anterior PVT (aPVT) neurons but predominantly inhibited posterior PVT (pPVT) neurons. Compared to pPVT with more inhibitory M2 receptors, aPVT expressed higher levels of all excitatory receptor subtypes including nicotinic α4, α7, and muscarinic M1 and M3. The ACh-induced excitation was mimicked by nicotine and antagonized by selective blockers for α4β2 and α7 nicotinic ACh receptor (nAChR) subtypes as well as selective antagonists for M1 and M3 muscarinic ACh receptors (mAChR). The ACh-induced inhibition was attenuated by selective M2 and M4 mAChR receptor antagonists. Furthermore, we found ACh increased the frequency of excitatory postsynaptic currents (EPSCs) on a majority of aPVT neurons but decreased EPSC frequency on a larger number of pPVT neurons. In addition, ACh caused an acute increase followed by a lasting reduction in inhibitory postsynaptic currents (IPSCs) on PVT neurons of both subregions. Together, these data suggest that multiple AChR subtypes coordinate a differential modulation of ACh on aPVT and pPVT neurons.