Line Model Research Articles

HERC5 downregulation in non-small cell lung cancer is associated with altered energy metabolism and metastasis

BackgroundMetastasis is the leading cause of cancer-related death in non-small cell lung cancer (NSCLC) patients. We previously showed that low HERC5 expression predicts early tumor dissemination and a dismal prognosis in NSCLC patients. Here, we performed functional studies to unravel the mechanism underlying the “metastasis-suppressor” effect of HERC5, with a focus on mitochondrial metabolism pathways.MethodsWe assessed cell proliferation, colony formation potential, anchorage-independent growth, migration, and wound healing in NSCLC cell line models with HERC5 overexpression (OE) or knockout (KO). To study early tumor cell dissemination, we used these cell line models in zebrafish experiments and performed intracardial injections in nude mice. Mass spectrometry (MS) was used to analyze protein changes in whole-cell extracts. Furthermore, electron microscopy (EM) imaging, cellular respiration, glycolytic activity, and lactate production were used to investigate the relationships with mitochondrial energy metabolism pathways.ResultsUsing different in vitro NSCLC cell line models, we showed that NSCLC cells with low HERC5 expression had increased malignant and invasive properties. Furthermore, two different in vivo models in zebrafish and a xenograft mouse model showed increased dissemination and metastasis formation (in particular in the brain). Functional enrichment clustering of MS data revealed an increase in mitochondrial proteins in vitro when HERC5 levels were high. Loss of HERC5 leads to an increased Warburg effect, leading to improved adaptation and survival under prolonged inhibition of oxidative phosphorylation.ConclusionsTaken together, these results indicate that low HERC5 expression increases the metastatic potential of NSCLC in vitro and in vivo. Furthermore, HERC5-induced proteomic changes influence mitochondrial pathways, ultimately leading to alterations in energy metabolism and demonstrating its role as a new potential metastasis suppressor gene.

Effects of low-frequency voltage on nonlinear standing wave excitation, plasma uniformity, and ion dynamics in dual-frequency asymmetric capacitive discharges

Abstract It is known that in very-high-frequency capacitively coupled plasmas (VHF CCPs), the higher harmonics generated by nonlinear sheath motion can enhance the standing wave effect (SWE), which can lead to center-peaked plasma density profiles. In this work, an improved nonlinear electromagnetic model incorporating a transmission line model, an electron momentum balance model, a bulk plasma model, a collisionless nonlinear numerical sheath model, and an ion Monte-Carlo collision (MCC) model is developed to study the effects of low-frequency (LF) voltage VL on the nonlinear standing wave excitation, plasma uniformity, and ion energy and angular distribution functions (IEDFs and IADFs) in dual-frequency (DF) asymmetric capacitive argon discharges at relatively low pressure of 3 Pa. The plasma diffusion in the radial direction and ion dynamics within the LF oscillating sheath are self-consistently considered. The LF voltage VL at 2 MHz varies from 0 to 700 V while the high-frequency (HF) voltage VH at 60 MHz is fixed at 100 V. Simulation results indicate that without the addition of an LF source (i.e. VL = 0 V), there are a considerable number of high-order harmonics with short wavelengths, leading to significant SWE and central peak in the radial plasma density profile. Nevertheless, the high-order harmonic excitations tend to be weakened and merely occur around the phase of the full LF sheath collapse due to a shorter characteristic damping time of the surface waves as VL increases. This, combined with increased surface wavelengths of both the driving frequency and the higher harmonics at a higher VL, leads to suppressed standing waves and improved plasma uniformity. Meanwhile, the simulations show that both the low and the high energy peaks of IEDF move towards higher energies, and the energy peak separation width ΔE becomes wider with the increase of VL. The IEDF at the radial center of the powered electrode exhibits a broader ΔE than that at the edge. For the IADF, an increased VL results in more ions incident on the electrode with a smaller deflection angle. Because of a thinner sheath and a higher sheath voltage at the electrode center, the peak value of IADF at the electrode center is greater than that at the edge.

Combination of Advanced Actuator Line/Disk Model and High-Order Unstructured Finite Volume Solver for Helicopter Rotors

In the research field of rotorcraft aerodynamics, there are two fundamental challenges: resolving the complex vortex structures in rotor wakes and representing the moving rotor blades in the ambient airflow. In this paper, we address the first challenge by utilizing a third-order unstructured finite volume solver, which exhibits lower numerical dissipation than its second-order counterpart. This allows for sufficient resolution of small vortex structures on relatively coarse meshes. With this flow solver, the second challenge is addressed by modeling each rotor as an actuator disk (i.e., the actuator disk model (ADM)) or modeling each blade as an actuator line (i.e., the actuator line model (ALM)). Both of the two models are equipped with an improved tip loss correction, which is introduced in detail in the methodology section. In the section of numerical experiments, the numerical convergence properties of the two types of solvers have been compared in the case of two-dimensional infinite wing. In addition, the relationship between the ALM and the lifting line theory is discussed in the cases of fixed-wing calculations. Another goal of these cases is to validate the tip loss correction presented. The validation of the ALM/ADM and comparisons of computational efficiency are also demonstrated in simulations involving both hover and forward flight rotors. It was found that the combination of the third-order finite volume solver and the ALM/ADM with the improved tip loss correction presents an efficient way of performing the aerodynamic analysis of rotor-induced downwash flow.

Telephone advice lines for adults with advanced illness and their family carers: a qualitative analysis and novel practical framework.

Telephone advice lines have been recommended internationally to support around-the-clock care for people living at home with advanced illness. While they undoubtedly support care, there is little evidence about what elements are needed for success. A national picture is needed to understand, improve and standardise service delivery/care. To explore telephone advice lines for people living at home with advanced illness across the four UK nations, and to construct a practical framework to improve services. A cross-national evaluation of telephone advice lines using structured qualitative interviews. A patient and public involvement workshop was conducted to refine the framework. Professionals with responsibilities for how palliative care services are delivered and/or funded at a local or regional level, were purposively sampled. Seventy-one interviews were conducted, covering 60 geographical areas. Five themes were identified. Availability: Ten advice line models were described. Variation led to confusion about who to call and when. Accessibility, awareness and promotion: It was assumed that patients/carers know who to call out-of-hours, but often they did not. Practicalities: Call handlers skills/expertise varied, which influenced how calls were managed. Possible responses ranged from signposting to organising home visits. Integration/continuity of care: Integration between care providers was limited by electronic medical records access/information sharing. Service structure/commissioning: Sustained funding was often an issue for charitably funded organisations. Our novel evidence-based practical framework could be transformative for service design/delivery, as it presents key considerations relating to the various elements of advice lines that may impact on the patient/carer experience.

Economic life cycle model for the cost of power grid line engineering project

Reasonable grid line engineering asset management is an important means for power companies to improve economic efficiency under the new electricity reform situation. Based on the whole life cycle theory, from the perspective of line asset acquisition, this paper put into use to the decommissioning of the whole process of cost perspective and analyzed the line project construction to decommissioning and scrapping of the whole process of the cost. Through the calculation of the whole life cycle of the smallest annual cost, to determine the line economic life, we constructed the whole life cycle of the grid line project economic cost framework, put forward the best decommissioning time of the grid line model, that is, the economic life model. A case study was conducted based on a project example from a certain power grid company in China. The research results of this paper can reduce the economic cost of operation and maintenance of power grid enterprises, improve the efficiency of input and output, and provide reference suggestions for optimizing the management of line engineering and other power assets of power grid companies.

Two-port quantum model of finite-length transmission lines coupled to lumped circuits

Two-port quantum model of finite-length transmission lines coupled to lumped circuits

Linc01588 deletion inhibits the malignant biological characteristics of hydroquinone-induced leukemic cells via miR-9-5p/SIRT1

Leukemia caused by environmental chemical pollutants has attracted great attention, the malignant leukemic transformation model of TK6 cells induced by hydroquinone (HQ) has been previously found in our team. However, the type of leukemia corresponding to this malignant transformed cell line model needs further study and interpretation. Furthermore, the molecular mechanism of malignant proliferation of leukemic cells induced by HQ remains unclear. This study is the first to reveal the expression of aberrant genes in leukemic cells of HQ-induced malignant transformation, which may correspond to chronic lymphocytic leukemia (CLL). The expression of Linc01588, a long non-coding RNA (lncRNA), was significantly up-regulated in CLL patients and leukemic cell line model which previously described. After gain-of-function assays and loss-of-function assays, feeble cell viability, severe apoptotic phenotype and the increased secretion of TNF-α were easily observed in malignant leukemic TK6 cells with Linc01588 deletion after HQ intervention. The tumors derived from malignant TK6 cells with Linc01588 deletion inoculated subcutaneously in nude mice were smaller than controls. In CLL and its cell line model, the expression of Linc01588 and miR-9–5p, miR-9–5p and SIRT1 were negative correlation respectively in CLL and cell line model, while the expression of Linc01588 and SIRT1 were positive correlation. The dual-luciferase reporter assay showed that Linc01588 & miR-9–5p, miR-9–5p & SIRT1 could bind directly, respectively. Furthermore, knockdown of miR-9–5p successfully rescued the severe apoptotic phenotype and the increased secretion of TNF-α caused by the Linc01588 deletion, the deletion of Linc01588 in human CLL cell line MEC-2 could also inhibit malignant biological characteristics, and the phenotype caused by the deletion of Linc01588 could also be rescued after overexpression of SIRT1. Moreover, the regulation of SIRT1 expression in HQ19 cells by Linc01588 and miR-9–5 P may be related to the Akt/NF-κB pathway. In brief, Linc01588 deletion inhibits the malignant biological characteristics of HQ-induced leukemic cells via miR-9–5p/SIRT1, and it is a novel and hopeful clue for the clinical targeted therapy of CLL.

Abstract LB427: UBA1 blockade induces homologous recombination deficiency and is synthetic lethal with PARP inhibition

Abstract Therapeutic strategies targeting DNA damage have revolutionized cancer treatment, yet their efficacy often comes at the cost of substantial adverse effects. The emergence of poly-ADP-ribose polymerase (PARP) inhibition as a targeted approach to disrupt DNA repair in tumors deficient in homologous recombination (HR) has shown promise. However, the limited response of most cancers to PARP inhibitors has posed a significant challenge. This study employs a genome-wide CRISPR knockout functional genomics screen, revealing that depletion of ubiquitin-activating enzyme E1 (UBA1) enhances sensitivity to PARP inhibition in HR-proficient ovarian cancer cells. Validation across different ovarian and breast cancer cell lineages, utilizing cell line and organoid models, establishes that pharmacological inhibition of UBA1, a pivotal player in ubiquitination, sensitizes cells to PARP inhibition and other DNA damaging therapies. In vivo experiments combining PARP and UBA1 inhibition demonstrate not only tolerability but also significant extension of survival in patient-derived xenografts compared to individual therapies. Mechanistic studies uncover that UBA1 inhibition not only impedes HR repair, sensitizing cells to PARP inhibition, but also induces widespread increases in PARylation, subsequently targeted by PARP inhibition. These findings underscore the multifaceted pathways through which UBA1 inhibition potentiates the efficacy of PARP inhibition. The demonstrated tolerability and substantial therapeutic gains establish the combined approach as a promising strategy to maximize the benefits of PARP inhibition in cancer therapy. Citation Format: Sharad Awasthi, Daniel J. McGrail, S. Stephen Yi, Nidhi Sahni. UBA1 blockade induces homologous recombination deficiency and is synthetic lethal with PARP inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB427.

Abstract LB403: Suppression of antigen presentation in pediatric rhabdomyosarcoma

Abstract Background: Rhabdomyosarcoma (RMS) is an aggressive soft tissue sarcoma that is diagnosed based on the tumor cell's resemblance to embryonic or poorly differentiated skeletal muscle progenitors. Multiple immunotherapy clinical trials have found no evidence of immune reactivity against RMS, which has been attributed to the relatively low tumor mutation burden in these cancers. However, we made an unexpected observation that many patient RMS tumors had a near absence of MHC-I expression, a key requisite for anti-tumor CD8+ T lymphocytes and many immunotherapies. In this study, we explore the etiology of low antigen presentation in RMS and systematically evaluate therapeutic interventions that sensitize RMS to immunotherapy. Methods: Surface MHC-I expression was evaluated on pediatric tumor cell lines (n=65) from five major pediatric tumor types. RNA-seq, whole exome and genome sequencing data were analyzed from RMS tumors (total of n=247) and iPSC models of skeletal muscle development (PMID: 32011235). Epigentic drugs targeting DNA methylation (decitabine), EZH2 (tazemetostat), Class I/IV HDACs (mocetinostat), and LSD1 (GSK-LSD1) were used at sub-cytotoxic doses on RMS cell lines (n=10) and analyzed for surface MHC-I expression with flow cytometry and gene expression. T cell cytotoxicity assays were performed using engineered human T cells expressing an optimized TCR which recognizes an HLA*A2:01 restricted peptide from PRAME. Results: RMS cell line models exhibited a range of MHC-I expression with most having low or undetectable MHC-I expression, similar to that of a known MHC-I negative tumor type MYCN-amplified neuroblastoma. Exome sequencing of RMS tumors showed no evidence of loss-of-function alterations in essential antigen processing and presentation (APP) pathway genes. Gene expression profiling by RNAseq and western blotting, however, revealed an absence of core APP genes in many RMS tumors and cell lines. We uncovered that during a specific window of embryonic myogenesis, APP is suppressed and that RMS tumors mimicking this state of development show the deepest loss of APP. Multiple classes of epigenetic drugs were evaluated to rescue APP in RMS. Treatment of RMS using a DNA demethylating agent or HDAC inhibitor exhibited the strongest MHC-I restoration across RMS models with no changes observed using LSD1 or EZH2 inhibitors. As a proof-of-concept, we demonstrate that targeting the RMS antigen PRAME using TCR therapy is enhanced by the inclusion of epigenetic therapy. Conclusions and Significance: We revealed that epigenetic suppression of APP is a new hallmark of RMS and normal embryonic skeletal muscle development. Pharmacological reversal of APP pathway downregulation could be achieved using drugs targeting DNA methylation or class I/IV HDACs, and enhanced T cell cytotoxicity against RMS when used in combination. Overall, these findings may explain the historical lack of RMS responsiveness to immunotherapy and unlocks new immunotherapy strategies against RMS that rely on targeting MHC-I antigens. Citation Format: David Milewski, Hsien-Chao Chou, Vineela Gangalapudi, Meijie Tian, Yong Kim, Young Song, Jun Wei, Javed Khan. Suppression of antigen presentation in pediatric rhabdomyosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB403.

Abstract LB022: Preclinical anti-tumorigenic evaluation of AXL targeting antibody-drug-conjugate in an adenoid cystic carcinoma cell line xenograft model

Abstract Adenoid cystic carcinoma (ACC) is a rare type of salivary gland cancer. Despite the many advances made in helping understand the disease behavior and characteristics, the prevailing challenge in managing patients with ACC is the lack of clinically approved systemic therapy drugs to successfully treat those with advanced recurrent or metastatic disease. There are multiple promising candidate agents being evaluated preclinically and in early phase clinical trials, yet an effective drug with consistent overall response remains to be identified. AXL is a member of the TAM family of tyrosine kinase receptors, and its overexpression and activation is generally associated with poor prognosis and chemoresistance in many cancer types. Previous reports show AXL is significantly upregulated in a subset of ACC tumors. Here, we evaluate the tumor growth inhibition effects of a newly developed antibody-drug-conjugate targeting AXL (ADCT-601) in a xenograft ACC cell line model. MDA-ACC-01 cells express high levels of cell surface AXL as confirmed by biochemical analysis. The tumor growth inhibition of MDA-ACC-01 cell xenograft was tested in a single-dose drug administration study. Immunodeficient mice bearing ACC tumors were either injected with vehicle (n=7) or with ADCT-601 at 0.5 (n=9) or 1.0 mg/kg (n=7). ADCT-601 treatment with either 0.5 mg/kg or 1.0 mg/kg resulted in complete response (CR) for all tumor implants. To reaffirm the drug’s efficacy, we challenged the vehicle-treated tumor, prior to exceeding the tumor volume endpoint, with a single-dose of ADCT-601 at 1.0 mg/kg. Strikingly, by day 6 post-treatment, tumor volume began to shrink and eventually resulted in CR except in all but one case. These results demonstrate ADCT-601 is an effective drug with significant anti-tumor activity in our preclinical models and is worthy of additional investigation to determine its translational potential. Citation Format: Joseph O. Humtsoe, Anita Pothukuchi, Hyunseok Kang, Patrick Ha. Preclinical anti-tumorigenic evaluation of AXL targeting antibody-drug-conjugate in an adenoid cystic carcinoma cell line xenograft model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB022.

Middle school mathematics teachers’ knowledge of integers

The current research aimed to unpack teachers’ knowledge of integers by investigating how they used the number line and counter models to represent the two meanings of division (i.e., partitive and measurement). The participants were three middle school mathematics teachers working in different cities in Türkiye. Data consisted of teachers’ written responses to an open-ended questionnaire consisting of four division operations and interviews conducted thereafter. Findings revealed that although two teachers could accurately model all division operations with the number line model, one teacher could neither provide a problem context nor a model displaying one of the division operations. For the counter model, only one teacher could accurately model all division operations in the questionnaire.

Sialyl-Tn serves as a potential therapeutic target for ovarian cancer

BackgroundOvarian cancer remains the deadliest of the gynecologic cancers in the United States. There have been limited advances in treatment strategies that have seen marked increases in overall survival. Thus, it is essential to continue developing and validating new treatment strategies and markers to identify patients who would benefit from the new strategy. In this report, we sought to further validate applications for a novel humanized anti-Sialyl Tn antibody-drug conjugate (anti-STn-ADC) in ovarian cancer.MethodsWe aimed to further test a humanized anti-STn-ADC in sialyl-Tn (STn) positive and negative ovarian cancer cell line, patient-derived organoid (PDO), and patient-derived xenograft (PDX) models. Furthermore, we sought to determine whether serum STn levels would reflect STn positivity in the tumor samples enabling us to identify patients that an anti-STn-ADC strategy would best serve. We developed a custom ELISA with high specificity and sensitivity, that was used to assess whether circulating STn levels would correlate with stage, progression-free survival, overall survival, and its value in augmenting CA-125 as a diagnostic. Lastly, we assessed whether the serum levels reflected what was observed via immunohistochemical analysis in a subset of tumor samples.ResultsOur in vitro experiments further define the specificity of the anti-STn-ADC. The ovarian cancer PDO, and PDX models provide additional support for an anti-STn-ADC-based strategy for targeting ovarian cancer. The custom serum ELISA was informative in potential triaging of patients with elevated levels of STn. However, it was not sensitive enough to add value to existing CA-125 levels for a diagnostic. While the ELISA identified non-serous ovarian tumors with low CA-125 levels, the sample numbers were too small to provide any confidence the STn ELISA would meaningfully add to CA-125 for diagnosis.ConclusionsOur preclinical data support the concept that an anti-STn-ADC may be a viable option for treating patients with elevated STn levels. Moreover, our STn-based ELISA could complement IHC in identifying patients with whom an anti-STn-based strategy might be more effective.

Establishing an Oxidative Stress Model in the Human Mesencephalic Cell Line (LUHMES): an in vitro study

Oxidative stress-caused neurodegenerative diseases, such as Alzheimer's, Parkinson's disease, and amyotrophic lateral sclerosis, are widely recognized as the most prevalent brain and central nervous system disorders. This is attributed to the vulnerability of neurons to oxidative stress within the body. Although substantial research has been performed on these diseases, it is extremely difficult to establish an oxidative stress model for brain tissues. In primary cultures, it is difficult to obtain neurons and the continuity of the culture is limited for in vitro cell line models. By providing valuable insights into the mechanisms of oxidative stress-induced neurodegenerative diseases, these in vitro models can aid in the development of effective treatment strategies. Here, we developed an in vitro oxidative stress model utilizing hydrogen peroxide on the LUHMES cell line. Our study evaluated the impact of this model on LUHMES cell viability and the equilibrium between oxidants and antioxidants by assaying total oxidant capacity (TOC) and total antioxidant capacity (TAC). Our results provided evidence of the oxidative effect of hydrogen peroxide in critical concentration and proved the efficacy of this model for further investigations.

The m6A methyltransferase METTL5 promotes neutrophil extracellular trap network release to regulate hepatocellular carcinoma progression.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, it has a poor prognosis due to its highly invasive and metastatic nature. Consequently, identifying effective prognostic markers and potential therapeutic targets has been extensively investigated. METTL5, an 18S rRNA methyltransferase, is abnormally high in HCC. But its biological function and prognostic significance in HCC remain largely unelucidated. This study aimed to investigate the role of METTL5 in HCC progression, and elucidate its possible molecular mechanisms in HCC via transcriptome sequencing, providing new insights for identifying new HCC prognostic markers and therapeutic targets. The METTL5 expression in HCC and paracancerous tissues was analyzed using HCC immunohistochemical microarrays and bioinformatic retrieval methods to correlate METTL5 with clinicopathological features and survival prognosis. We constructed a METTL5 knockdown hepatocellular carcinoma cell line model and an animal model to determine the effect of METTL5 on hepatocellular carcinoma progression. Subsequently, RNA sequencing was performed to analyze the molecular mechanism of METTL5 in HCC based on the sequencing results, and relevant experiments were performed to verify it. We found that METTL5 expression was elevated in hepatocellular carcinoma tissues and correlated with poor patient prognosis, and in the analysis of clinicopathological features showed a correlation with TNM staging. In hepatocellular carcinoma cell lines with knockdown of METTL5, the malignant biological behavior was significantly reduced both invitro and invivo. Based on the sequencing results as well as the results of GO functional enrichment analysis and KEGG pathway enrichment analysis, we found that METTL5 could promote the generation and release of neutrophil extracellular capture network (NETs) and might further accelerate the progression of HCC. The m6A methyltransferase METTL5 is overexpressed in hepatocellular carcinoma (HCC) and correlates with poor prognosis. METTL5 accelerates malignant progression of HCC by promoting generation and release of the neutrophil extracellular traps (NETs) network, providing new insights for clinical biomarkers and immunotherapeutic targets in HCC prognosis.

Fast computation of the observer motion effects induced on monopole frequency spectra for tabulated functions

Context. Various methods have been studied to compute the boosting effects produced by the observer peculiar motion, which modifies and transfers the isotropic monopole frequency spectrum of the cosmic background radiation to higher multipoles. Explicit analytical solutions for the spherical harmonic expansion coefficients were already presented and applied to different types of background spectrum, strongly alleviating the computational effort needed for accurate theoretical predictions. The frequency spectra at higher multipoles are inherently led by higher-order derivatives of the monopole spectrum. Provided that it can be well described by analytic or semi-analytic functions, the computation of its transfer is not affected by numerical instabilities when evaluated at the required level of numerical accuracy. Instead, monopole frequency spectra described by tabulated functions are computed with a relatively poor frequency resolution in comparison with the Doppler shift, which necessitates interpolation of the tabular representation. The spectra are also affected by uncertainties related to intrinsic inaccuracies in the modelling or in the related observational data as well as to limited accuracy in their numerical computation. These uncertainties propagate and increase with the derivative order, possibly preventing the trustworthy computation of the transfer to higher multipoles and of the observed monopole. Aims. We study methods to filter the original function or its derivatives and the multipole spectra, to mitigate numerical instabilities, and to derive reliable predictions of the harmonic coefficients for different cosmic background models. Methods. From the analytical solutions, and assuming that the monopole spectrum can be expanded in Taylor’s series, we derive explicit expressions for the harmonic coefficients up to the multipole ℓmax = 6 in terms of monopole spectrum derivatives. We then consider different low-pass filters: prefiltering in Fourier space of the tabular representation; filtering in both real and Fourier space of the numerical derivatives; interpolation approaches; and a dedicated method based on amplification and deamplification of the boosted signal. We study the quality of these methods when applied to suitable analytical approximations of the tabulated functions, possibly polluted with simulated noise. These methods are then applied to the tabulations. Results. We consider two very different types of monopole spectra superimposed to the cosmic microwave background: the (smooth) extragalactic source microwave background signal from radio-loud active galactic nuclei and the (feature-rich) redshifted 21 cm line, and present our results in terms of spherical harmonic coefficients. The direct prediction of these coefficients can be noisy at ℓ > 1 or, depending on the uncertainty level, even at ℓ ≤ 1. Without assuming a functional form for the extragalactic background spectrum, the Gaussian prefiltering coupled to the sequential real-space filtering of derivatives allows us to derive accurate predictions up to ℓ ∼ 6, while a log–log polynomial representation, which is appropriate over several decades, gives accurate solutions at any ℓ. Instead, it is difficult to characterise the 21 cm line model variety, and so it is relevant to work without assumptions about the underlying function. Typically, the prefiltering provides accurate predictions up to ℓ ≃ 3 or 4, while the further sequential filtering of the derivatives or the boosting amplification and deamplification method improves the results up to ℓ = 4, while also allowing reasonable estimations of the spectrum at higher ℓ. Conclusions. The proposed methods can significantly extend the range of realistic cosmic background models manageable with a fast computation, beyond the cases characterised a priori by analytical or semi-analytical functions. These methods require only an affordable increase in computation time compared to the direct calculation via simple interpolation.

504 Dihydroxyacetone, a combustion of electronic cigarettes, promotes cardiac-specific injury through metabolic and mitochondrial imbalances

OBJECTIVES/GOALS: Electronic cigarettes have become increasingly popular, with various combustion products generated in the process. Dihydroxyacetone (DHA), a carbohydrate made during the heating process. Exposures may reach high micromolar to low millimolar doses of DHA per day and no studies have been done to understand the effects of DHA in the heart. METHODS/STUDY POPULATION: Here, we examine if DHA contributes to these using rat cardiomyocytes, H9c2 cells, and rat cardiac tissues to DHA evaluating metabolic and mitochondrial effects. Using the cells, we will investigate metabolic and mitochondrial pathways using Seahorse, protein expression changes in nutrient sensing pathways, and understand dose-dependent effects of DHA in the heart. Metabolite pools will also be evaluated to understand the changes promoted by DHA. Oxidative stress as previously observed in other cell models will also be measured. Key findings in the cardiac cells will be investigated in the cardiac tissues exposed to DHA. RESULTS/ANTICIPATED RESULTS: We have previously shown DHA induces oxidative stress, metabolic changes, and mitochondrial dysfunction in various cell line models. Interestingly, these effects are highly cell-type dependent. E-cigarettes are known to have toxic cardiac effects, including arterial stiffness, endothelial dysfunction, vascular injury, and oxidative stress. Changes in glycolytic, fatty acid synthesis, and the citric acid cycle enzymes and metabolites were found in the H9c2 cells. We also observed increased mitochondrial ROS and fuel changes due to DHA exposure. In DHA exposed cardiac tissues, we observed oxidative stress and mitochondrial fission and fusion dynamics altered. DISCUSSION/SIGNIFICANCE: These data suggest further study at physiologically relevant doses is warranted to understand how DHA inhaled impacts the long-term health of vapers. As well as the regulation of DHA in e-cigarettes as it has been deemed as safe for topical applications and warned against inhalation.

Immunomodulatory potential of rapamycin-loaded mesoporous silica nanoparticles: pore size-dependent drug loading, release, and in vitro cellular responses.

Rapamycin is a potent immunosuppressive drug that has been recently proposed for a wide range of applications beyond its current clinical use. For some of these proposed applications, encapsulation in nanoparticles is key to ensure therapeutic efficacy and safety. In this work, we evaluate the effect of pore size on mesoporous silica nanoparticles (MSN) as rapamycin nanocarriers. The successful preparation of MSN with 4 different pore sizes was confirmed by dynamic light scattering, zeta potential, transmission electron microscopy and N2 adsorption. In these materials, rapamycin loading was pore size-dependent, with smaller pore MSN exhibiting greater loading capacity. Release studies showed sustained drug release from all MSN types, with larger pore MSN presenting faster release kinetics. In vitro experiments using the murine dendritic cell (DC) line model DC2.4 showed that pore size influenced the biological performance of MSN. MSN with smaller pore sizes presented larger nanoparticle uptake by DC2.4 cells, but were also associated with slightly larger cytotoxicity. Further evaluation of DC2.4 cells incubated with rapamycin-loaded MSN also demonstrated a significant effect of MSN pore size on their immunological response. Notably, the combination of rapamycin-loaded MSN with an inflammatory stimulus (lipopolysaccharide, LPS) led to changes in the expression of DC activation markers (CD40 and CD83) and in the production of the proinflammatory cytokine TNF-α compared to LPS-treated DC without nanoparticles. Smaller-pored MSN induced more substantial reductions in CD40 expression while eliciting increased CD83 expression, indicating potential immunomodulatory effects. These findings highlight the critical role of MSN pore size in modulating rapamycin loading, release kinetics, cellular uptake, and subsequent immunomodulatory responses.

Fourteen undescribed steroidal saponins from Solanum capsicoides leaves and their neuroprotective effects

A total of 14 previously undescribed steroidal saponins named capsicsaponins A-N were isolated from the leaves of Solanum capsicoides, encompassing various types, including cholesterol derivatives and pseudospirostanol saponins. The structures of all compounds were determined through comprehensive analysis of spectroscopic data (1D NMR and 2D NMR), along with physicochemical analysis methods (acid hydrolysis, OR, and UV). Moreover, in the H2O2-induced pheochromocytoma cell line model, compounds 1-14 were screened for their neuroprotective effects on cells. The bioassay results demonstrated compounds 8-14 were able to revive cell viability compared to the positive control edaravone. The damage neuroprotection of the most active compound was further explored.

Revitalizing Oxygen Reduction Reactivity of Composite Oxide Electrodes via Electrochemically Deposited PrOx Nanocatalysts.

Solid oxide fuel cells that operate at intermediate temperatures require efficient catalysts to enhance the inherently poor electrochemical activity of the composite electrodes. Here, a simple and practical electrochemical deposition method is presented for fabricating a PrOx overlayer on lanthanum strontium manganite-yttria-stabilized zirconia (LSM-YSZ) composite electrodes. The method requires less than four minutes for completion and can be carried out under at ambient temperature and pressure. Crucially, the treatment significantly improves the electrode's performance without requiring heat treatment or other supplementary processes. The PrOx-coated LSM-YSZ electrode exhibits an 89% decrease in polarization resistance at 650°C (compared to an untreated electrode), maintaining a tenfold reduction after ≈400h. Transmission line model analysis using impedance spectra confirms how PrOx coating improved the oxygen reduction reaction activity. Further, tests with anode-supported single cells reveal an outstanding peak power density compared to those of other LSM-YSZ-based cathodes (e.g., 418mWcm-2 at 650°C). Furthermore, it is demonstrated that multicomponent coating, such as (Pr,Ce)Ox, can also be obtained with this method. Overall, the observations offer a promising route for the development of high-performance solid oxide fuel cells.

Design of Energy Efficient & Sustainable Buildings for ISRO Data Centers Using E-Quest

Saving energy is the same as producing new energy. Sustainable development has elevated the importance of energy performance and energy efficiency. Energy efficiency is a crucial part of sustainability, which includes all sectors of the economy from agriculture to manufacturing to services to infrastructure, including buildings used for business and research activities. Developing countries face unique difficulties in achieving sustainability because they must balance two phenomena that are diametrically opposed to one another: rapid economic growth and long-term prosperity. Commercial, information technology (IT), real estate development and construction are important economic drivers. Energy is, of course, a crucial consideration in these fields. In this research, we propose a design strategy for improving energy efficiency in an ISRO Data center by using sustainable green building technology [1]. This paper discusses the initiatives of ISRO in this direction. As maximum operation & maintenance cost of the buildings is due to electrical energy, this study shows how to design the most energy efficient institutional buildings for IMGEOS (Integrated Multi mission Ground Segment for Earth Observation satellites) of ISRO, Hyderabad. The research work discuses about the strategies / techniques to be considered since the planning & designing stage of the building infrastructure itself. This research gives the complete details of the materials with their detailed technical specifications and the methodology to be adopted, for achieving the required energy efficiency level in the data centers / similar IT infrastructure buildings. The research work also explains the process of finding & selecting the most energy efficient model by using different combination of green building materials, with respect to the base line model of ASHRAE using the e-Quest building simulation software and the results obtained thereof. Thus, this detailed research study investigates the process of designing the energy efficient data center buildings for ISRO in the context of emerging countries.