Enriched Targets Research Articles

Sustainable production of radionuclidically pure antimony-119

BackgroundRadiopharmaceutical therapy (RPT) uses radionuclides that decay via one of three therapeutically relevant decay modes (alpha, beta, and internal conversion (IC) / Auger electron (AE) emission) to deliver short range, highly damaging radiation inside of diseased cells, maintaining localized dose distribution and sparing healthy cells. Antimony-119 (119Sb, t1/2 = 38.19 h, EC = 100%) is one such IC/AE emitting radionuclide, previously limited to in silico computational investigation due to barriers in production, chemical separation, and chelation. A theranostic (therapeutic/diagnostic) pair can be formed with 119Sb’s radioisotopic imaging analogue 117Sb (t1/2 = 2.80 h, Eγ = 158.6 keV, Iγ = 85.9%, β+ = 262.4 keV, Iβ+ = 1.81%).ResultsWithin, we report techniques for sustainable and cost-effective production of pre-clinical quality and quantity, radionuclidically pure 119Sb and 117Sb, novel low energy photon measurement techniques for 119Sb activity determination, and physical yields for various tin target isotopic enrichments and thicknesses using (p, n) and (d, n) nuclear reactions. Additionally, we present a two-column separation providing a radioantimony yield of 73.1% ± 6.9% (N = 3) and tin separation factor of (6.8 ± 5.5) x 105 (N = 3). Apparent molar activity measurements for deuteron produced 117Sb using the chelator TREN-CAM were measured at 42.4 ± 25 MBq 117Sb/µmol (1.14 ± 0.68 mCi/µmol), and we recovered enriched 119Sn target material at a recycling efficiency of 80.2% ± 5.5% (N = 6) with losses of 11.6 mg ± 0.8 mg (N = 6) per production.ConclusionWe report significant steps in overcoming barriers in 119Sb production, chemical isolation and purification, enriched target material recycling, and chelation, helping promote accessibility and application of this promising therapeutic radionuclide. We describe a method for 119Sb activity measurement using its low energy gamma (23.87 keV), negating the need for attenuation correction. Finally, we report the largest yet-measured 119Sb production yields using proton and deuteron irradiation of natural and enriched targets and radioisotopic purity > 99.8% at end of purification.

Identification of molecular targets of Hypericumperforatum in blood for major depressive disorder: a machine-learning pharmacological study

BackgroundMajor depressive disorder (MDD) is one of the most common psychiatric disorders worldwide. Hypericumperforatum (HP) is a traditional herb that has been shown to have antidepressant effects, but its mechanism is unclear. This study aims to identify the molecular targets of HP for the treatment of MDD.MethodsWe performed differential analysis and weighted gene co-expression network analysis (WGCNA) with blood mRNA expression cohort of MDD and healthy control to identify DEGs and significant module genes (gene list 1). Three databases, CTD, DisGeNET, and GeneCards, were used to retrieve MDD-related gene intersections to obtain MDD-predicted targets (gene list 2). The validated targets were retrieved from the TCMSP database (gene list 3). Based on these three gene lists, 13 key pathways were identified. The PPI network was constructed by extracting the intersection of genes and HP-validated targets on all key pathways. Key therapeutic targets were obtained using MCODE and machine learning (LASSO, SVM-RFE). Clinical diagnostic assessments (Nomogram, Correlation, Intergroup expression), and gene set enrichment analysis (GSEA) were performed for the key targets. In addition, immune cell analysis was performed on the blood mRNA expression cohort of MDD to explore the association between the key targets and immune cells. Finally, molecular docking prediction was performed for the targets of HP active ingredients on MDD.ResultsDifferential expression analysis and WGCNA module analysis yielded 933 potential targets for MDD. Three disease databases were intersected with 982 MDD-predicted targets. The TCMSP retrieved 275 valid targets for HP. Separate enrichment analysis intersected 13 key pathways. Five key targets (AKT1, MAPK1, MYC, EGF, HSP90AA1) were finally screened based on all enriched genes and HP valid targets. Combined with the signaling pathway and immune cell analysis suggested the effect of peripheral immunity on MDD and the important role of neutrophils in immune inflammation. Finally, the binding of HP active ingredients (quercetin, kaempferol, and luteolin) and all 5 key targets were predicted based on molecular docking.ConclusionsThe active constituents of Hypericumperforatum can act on MDD and key targets and pathways of this action were identified.

The production and separation of 161Tb with high specific activity at the University of Utah

Targeted radiotherapy (TRT) is an increasingly prominent area of research in nuclear medicine, particularly in the context of treating cancerous tumors. One radionuclide of considerable interest for TRT is terbium-161 (t1/2 = 6.95 days), which undergoes beta emission and shares similar decay properties as 177Lu (FDA-approved as LUTATHERA® and PLUVICTO®). Besides beta emission, 161Tb also emits a significant number of conversion and Auger electrons further enhancing its therapeutic potential. Terbium-161 can be produced using nuclear reactors through an indirect neutron capture reaction, G64160dn,γG64161d→3.66min,β−T65161b, from 160Gd targets. However, a key challenge in utilizing 161Tb for TRT lies in effectively separating target and product materials to attain high specific activity for radiolabeling. Here, we detail the production of no-carrier added 161Tb using low flux research reactors (mean thermal (<0.625 eV) neutron flux: 1.356×1012n∙cm−2∙s−1) like the University of Utah TRIGA Reactor, using enriched 160Gd2O3 targets (1.5 ± 0.3 μCi of 161Tb per mg of 160Gd target per hour of irradiation). We also developed a separation technique based on cation exchange and extraction chromatography, suitable for mCi level irradiations with targets exceeding 200 mg. In a simulated full-scale irradiation, 161Tb was successfully isolated from large mass targets using cation exchange (AG 50W-X8, with 2-hydroxyisobutyric acid at 70 mM, pH 4.75) and extraction chromatography (LN Resin, 0.5–0.75 M HNO3) methods. This resulted in high apparent molar activities of [161Tb]Tb-DOTA (113 ± 3 MBq/nmol), demonstrating high purity 161Tb relevant for potential future preclinical applications.

Development of Target Holder for the First Experiment on an Alternative Route for 64Cu Radioisotope Production by Secondary Neutron Irradiation of a 64ZnO Target

The target holder, as part of the target system for cyclotron-based radioisotope production, plays a crucial role in successful radioisotope production. The target holder has to be designed and developed so that it will not deform or melt should a beam of energetic particles irradiate the target. In this work, we develop and test a target holder for 64Cu radioisotope production. The thermal distribution and structural analysis are simulated using ANSYS software. Based on the ANSYS simulation results, a maximum temperature of 84°C occurred on the titanium foil, while the maximum temperature in the target holder body was 35.6°C when an 11-MeV proton beam with a beam current of 25 μA was bombarded on the target holder. We successfully test the target holder, and for the first time, we experimentally produce a 64Cu radioisotope by secondary neutron irradiation of the 64ZnO target. Using 11-MeV protons with a proton beam current of 25 μA incident on a 1-mm Ti foil for 5 min, we were able to generate secondary neutrons, and then the secondary neutrons irradiated 1 g of the enriched 64ZnO target. Copper-64 produced from the 64Zn(n,p)64Cu nuclear reaction was eventually detected using a portable gamma spectrometer, and its radioactivity was measured using a dose calibrator. For the first time, this experimental study confirmed that as much as 48.8 ± 6.2 MBq/μAh radioactivity of 64Cu was produced with no observed radioactive impurities.

Combining rVAR2 and Anti-EpCAM to Increase the Capture Efficiency of Non-Small-Cell Lung Cancer Cell Lines in the Flow Enrichment Target Capture Halbach (FETCH) Magnetic Separation System.

Circulating tumor cells (CTCs) are detected in approximately 30% of metastatic non-small-cell lung cancer (NSCLC) cases using the CellSearch system, which relies on EpCAM immunomagnetic enrichment and Cytokeratin detection. This study evaluated the effectiveness of immunomagnetic enrichment targeting oncofetal chondroitin sulfate (ofCS) using recombinant VAR2CSA proteins (rVAR2) to improve the recovery of different NSCLC cell lines spiked into lysed blood samples. Four NSCLC cell lines-NCI-H1563, A549, NCI-H1792, and NCI-H661-were used to assess capture efficiency. The results demonstrated that the combined use of anti-EpCAM antibody and rVAR2 significantly enhanced the capture efficiency to an average of 88.2% compared with 40.6% when using only anti-EpCAM and 56.6% when using only rVAR2. These findings suggest that a dual-marker approach using anti-EpCAM and rVAR2 can provide a more robust and sensitive method for CTC enrichment in NSCLC, potentially leading to better diagnostic and prognostic outcomes.

A novel interaction between RNA m6A methyltransferase METTL3 and RREB1

Regulation of gene expression is achieved through the modulation of regulatory inputs both pre- and post-transcriptionally. Methyltransferase-like 3 (METTL3) is a key player in pre-mRNA processing, actively catalyzing N6-methyladenosine (m6A). Among the most enriched mRNA targets of METTL3 is the Ras Responsive Element Binding Protein 1 (RREB1), a transcription factor which functions to govern cell fate, proliferation and DNA repair. Here, we show a novel interaction between METTL3 and RREB1. Further examination of this interaction indicates that METTL3's N-terminus is the primary interacting domain. Our findings uncover a novel interacting partner of METTL3, providing further insights into METTL3's regulatory network.

A muon beam monitor for the CHNet-MAXI experiment at the RIKEN-RAL laboratory

The CHNet-MAXI experiment (Muonic Atom X-ray and prompt gamma spectroscopy for Isotopic analysis for cultural heritage) is an INFN Cultural Heritage program funded by Scientific Commission V, aiming at selecting the best figures of merit in the isotopic analysis of lead by means of muonic atom emission. The experiment will be deployed at RIKEN-RAL facility ISIS-STFC (UK), providing a pulsed muon beams with momenta between 15 and 90 MeV/c. The CHNet-MAXI apparatus will be mainly based on a 9 HPGe detectors array and innovative scintillator detectors read-out by SiPMs for the detection of the characteristic X and gamma rays emitted by isotopically enriched targets of Pb 204, 206, 207 and 208, after negative muon irradiation at the RIKEN-RAL beamlines. A muon beam hodoscope will be used as a beam x/y profiler and as a beam intensity monitor. This detector has been designed to provide a few mm spacial resolution, and it is based on 3 mm thick scintillating fibers, each read-out by a single SiPM at one edge. The setup is meant to be portable thanks to desktop HV and innovative open-FPGA DAQ modules. All the phases of the design, assembly, and testing of the CHNet-MAXI hodoscope are illustrated. Finally, first operation tests of this new beam monitor are shown.

Impact of the near-physiological temperature on the in vitro maturation of bovine oocytes: A comparative proteomic approach

In vivo, the temperature inside preovulatory follicles of cows is approximately 1 °C lower than rectal temperature. However, standard bovine oocyte in vitro maturation (IVM) protocols use 38.5 °C based on rectal temperature. This study evaluated the effect of reducing IVM temperature to 37.5 °C on the proteomic profile of oocytes compared to the routine 38.5 °C. Nuclear maturation rate and cumulus cell (CC) expansion (30 COCs per group, 21 replicates) were assessed by observing the first polar body and using a subjective scoring method (0–4). Total nitrite concentrations in the culture medium were measured using the Griess method. Differential proteomics was performed using LC-MS/MS on pooled oocyte samples (500 matured oocytes per group, three replicates), followed by gene ontology enrichment, protein-protein interaction, and putative miRNA target analyses. No significant differences were observed between the groups in nuclear maturation, CC expansion, or nitrite concentration (P > 0.05). A total of 806 proteins were identified, with 7 up-regulated and 12 down-regulated in the treatment group compared to the control. Additionally, 12 proteins were unique to the control group, and 8 were unique to the treatment group. IVM at 37.5 °C resulted in the upregulation of proteins involved in protein folding and GTP binding, and the downregulation of enzymes with oxidoreductase activity and proteins involved in cytoskeletal fiber formation. Furthermore, 43 bovine miRNAs potentially regulating these genes (DES, HMOX2, KRT75, FARSA, IDH2, CARHSP1) were identified. We conclude that IVM of bovine oocytes at 37.5 °C induces significant proteomic changes without impacting nuclear maturation, cumulus cell expansion, or nitrite concentration in the IVM medium.

UNC13B regulates the sensitivity of Wilms' tumor cells to doxorubicin by modulating lysosomes.

Wilms' tumor is a malignant neoplasm where current medical advancements have significantly improved survival rates; however, challenges persist such as the resistance of the tumor to chemotherapy drugs like doxorubicin. This necessitates higher dosages, leading to decreased sensitivity. However, using high doses of doxorubicin can have late effects on the heart. Unc-13 homolog B (UNC13B) may be involved in the drug resistance in several tumors, yet its role in modulating drug sensitivity in Wilms' tumor remains unexplored. UNC13B levels were quantified using reverse transcription-qPCR and Western blotting. The half-maximal inhibitory concentration for doxorubicin, vincristine, and actinomycin-D was determined using CCK-8 assays. Cell cycle and apoptosis were analyzed using flow cytometry, and lysosomal changes were observed using Lyso-Tracker staining. The present study initially evaluated UNC13B expression levels in the Wilms' tumor 17.94 cell line. Additionally, through short hairpin RNA-mediated knockdown, changes in doxorubicin sensitivity in 17.94 Wilms' tumor cells were assessed. Concurrently, preliminary investigations into the role of UNC13B in regulating lysosomes was performed, revealing a significant positive association between UNC13B levels and lysosome formation in the 17.94 cell line. Lysosomes likely serve a role in the sensitivity of Wilms' tumor cell lines to drugs. Elevated UNC13B expression was observed in the 17.94 Wilms' tumor cell line compared to normal kidney cells. UNC13B knockdown also resulted in increased apoptosis levels upon doxorubicin treatment. Immunofluorescence revealed UNC13B localization within cellular vesicles, and its knockdown significantly decreased lysosome levels. Overall, the findings of the present study demonstrate that UNC13B regulates the sensitivity of the Wilms' tumor 17.94 cell line to doxorubicin by modulating lysosome formation within cells. The results suggest that UNC13B is likely an enriched target involved in lysosomal regulation in certain tumors, offering a new approach for optimizing chemotherapy in Wilms' tumor and other cancers with high UNC13B expression.

Boosting the odour perception of trace compounds exploiting three-dimensional GC-O with MS/FID simultaneous detection

ABSTRACT The study describes the effectiveness of a 3D GC system coupled with olfactometric detection and simultaneous MS and FID (3DGC-O-MS/FID) to improve the detection of odorants. Mega-bore columns were used to ensure a sample capacity able to enhance the odour perception. The heart-cut approach allowed to overcome the low efficiency of wide-bore columns, granting to purify enriched target fractions before olfactometry detection. After the third GC dimension, the isolated target peaks were eluted to the olfactometric port allowing the odour to last multiple seconds, making the panelist’s perception easier and more comfortable. The simultaneous FID/MS detection after the third GC dimension allowed to match specific odours to target molecules and their concentration in the sample. The 3DGC-O-MS/FID system boosts odour perception of trace components compared to conventional GC-O approaches. A reduced detection threshold of mg/kg levels was obtained compared to the mg/kg levels typical of monodimensional GC-O.

Excitation functions of72Ge(p,xn)72,71As reactions from threshold up to 45 MeV for production of the non-standard positron emitter 72As

Nuclear reaction cross sections for the formation of 72As and 71As in proton-induced reactions on enriched 72Ge targets were measured up to 45 MeV utilizing three different cyclotrons at the Forschungszentrum Jülich. The stacked-thin sample activation technique in combination with high-resolution γ-ray spectrometry was used. The major γ-ray peaks of 72As and 71As formed via the 72Ge(p,n)72As and 72Ge(p,2n)71As reactions, respectively, were analyzed. The incident proton energy and flux on a foil were determined using several monitor reactions. Based on integrated counts, irradiation data and the nuclear decay data, the reaction cross sections were measured. All data describe the first measurements. Theoretical nuclear model calculations were then carried out by using the codes TALYS 1.96, EMPIRE 3.2 and ALICE-IPPE. A very good agreement between the measured data and calculated values was found. The new data enabled us to calculate the thick target yields and estimate the radionuclidic impurities for a given energy range. Over the optimum energy range Ep = 14 → 7 MeV, the calculated thick target yield of 72As amounts to 272 MBq/μAh with no 71As impurity at all. The 72Ge(p,n)72As reaction on the enriched 72Ge is thus very suitable for clinical scale production of 72As at a medical cyclotron.

Upgrading low-concentration oxygen-bearing coal bed methane by dual-reflux vacuum swing adsorption

Fugitive methane (CH4) is a typical by-product of mining processes, which is commonly known as coal bed methane (CBM) or coal mine gas (CMG). The capture of these CH4 gases can simultaneously avoid greenhouse gas emissions and provide extra energy benefits. However, the explosion risk of low-concentration CBM (CH4 molar fraction ≤ 30%) requires strictly safe operating protocols to conduct the capture process. Dual reflux vacuum swing adsorption (DR-VSA) is a promising candidate with a vacuum operating condition which can lower the explosion risk and simultaneously reach CH4 enrichment and O2 removal targets in product and effluent streams. Herein, a low-concentration oxygen-bearing CBM (20% CH4, 16% O2 and 64% N2) can be upgraded to 69.7 mol% in the product gas while ensuring an effluent concentration of 2.5 mol% by the DR-VSA cycle using ionic liquidic zeolites (ILZ) as the adsorbents. A rigorous safety analysis has been conducted to investigate the explosion risk in the adsorption column and product tank, suggesting that the DR-VSA process is a safe technology for upgrading low-concentration oxygen-bearing methane.

Reactive Oxygen Species Responsive Multifunctional Fusion Extracellular Nanovesicles: Prospective Treatments for Acute Heart Transplant Rejection.

Heart transplantation offers life-saving treatment for patients with end-stage heart failure; however, ischemia-reperfusion injury (IRI) and subsequent immune responses remain significant challenges. Current therapies primarily target adaptive immunity, with limited options available for addressing IRI and innate immune activation. Although plant-derived vesicle-like nanoparticles show promise in managing diseases, their application in organ transplantation complications is unexplored. Here, this work develops a novel reactive oxygen species (ROS)-responsive multifunctional fusion extracellular nanovesicles carrying rapamycin (FNVs@RAPA) to address early IRI and Ly6C+Ly6G- inflammatory macrophage-mediated rejection in heart transplantation. The FNVs comprise Exocarpium Citri grandis-derived extracellular nanovesicles with anti-inflammatory and antioxidant properties, and mesenchymal stem cell membrane-derived nanovesicles expressing calreticulin with macrophage-targeting ability. A novel ROS-responsive bio-orthogonal chemistry approach facilitates the active targeting delivery of FNVs@RAPA to the heart graft site, effectively alleviating IRI and promoting the polarization of Ly6C+Ly6G- inflammatory macrophages toward an anti-inflammatory phenotype. Hence, FNVs@RAPA represents a promising therapeutic approach for mitigating early transplantation complications and immune rejection. The fusion-targeted delivery strategy offers superior heart graft site enrichment and macrophage-specific targeting, promising improved transplant outcomes.

Formation of medical radioisotope 111In in photonuclear reactions

The possibility of photonuclear production of 111In radioisotope has been investigated. The enriched target 112Sn was irradiated at the linear electron accelerator LUE-75 of A. Alikhanian National Science Laboratory (Yerevan, Armenia) at the bremsstrahlung endpoint energy Eγmax = 55 MeV. The cross section per equivalent quantum for reactions 112Sn(γ,x)111In, 112Sn(γ,n)111Sn, 112Sn(γ,2n)110Sn, 112Sn(γ,3n)109Sn,112 Sn(γ,pn)110mIn, 112Sn(γ,pn)110g110In, 112Sn(γ,p2n)109In have been measured via the method of activation and off-line γ-ray spectrometric technique. The cross section per equivalent quantum of the 111In in photonuclear reaction was compared with its cross section in proton induced reaction on cadmium targets and other possible 111In production routes. It is shown that the photonuclear method can be used for the production of 111In.

Biomarkers in pursuit of precision medicine for acute kidney injury: hard to get rid of customs.

Traditional acute kidney injury (AKI) classifications, which are centered around semi-anatomical lines, can no longer capture the complexity of AKI. By employing strategies to identify predictive and prognostic enrichment targets, experts could gain a deeper comprehension of AKI's pathophysiology, allowing for the development of treatment-specific targets and enhancing individualized care. Subphenotyping, which is enriched with AKI biomarkers, holds insights into distinct risk profiles and tailored treatment strategies that redefine AKI and contribute to improved clinical management. The utilization of biomarkers such as N-acetyl-β-D-glucosaminidase, tissue inhibitor of metalloprotease-2·insulin-like growth factor-binding protein 7, kidney injury molecule-1, and liver fatty acid-binding protein garnered significant attention as a means to predict subclinical AKI. Novel biomarkers offer promise in predicting persistent AKI, with urinary motif chemokine ligand 14 displaying significant sensitivity and specificity. Furthermore, they serve as predictive markers for weaning patients from acute dialysis and offer valuable insights into distinct AKI subgroups. The proposed management of AKI, which is encapsulated in a structured flowchart, bridges the gap between research and clinical practice. It streamlines the utilization of biomarkers and subphenotyping, promising a future in which AKI is swiftly identified and managed with unprecedented precision. Incorporating kidney biomarkers into strategies for early AKI detection and the initiation of AKI care bundles has proven to be more effective than using care bundles without these novel biomarkers. This comprehensive approach represents a significant stride toward precision medicine, enabling the identification of high-risk subphenotypes in patients with AKI.

Separation of terbium as a first step towards high purity terbium-161 for medical applications.

Terbium-161 is a medical radiolanthanide that has a beta decay energy and half-life similar to that of lutetium-177, which makes it a promising alternative for therapeutic purposes. The production route using an enriched gadolinium-160 target necessitates the purification of terbium-161 from the untransmuted target material as well as from its stable decay product, dysprosium-161. The separation of neighbouring lanthanides is challenging due to their similar chemical properties and prominent trivalent oxidation states. In this work, the aim is to change the oxidation state of terbium, resulting in the altering of chemical properties that ease the intragroup separation. To this end, a novel separation method is investigated, involving the electrochemical oxidation of terbium (3+) to terbium (4+) followed by anion exchange chromatography. The electrolysis conditions are set to the highest achievable conversion rate, followed by a dilution step during which the pH and electrolyte concentration are slightly lowered to obtain conditions that are compatible with the separation method. XAS analysis is done to characterize the carbonato complex of both oxidation states and to further elucidate the separation mechanism. The results show that the separation approach of combining electrochemical oxidation with anion exchange chromatography is promising for the purification of 161Tb for medical use.

Converging peripheral blood microRNA profiles in Parkinson's disease and progressive supranuclear palsy.

MicroRNAs act via targeted suppression of messenger RNA translation in the DNA-RNA-protein axis. The dysregulation of microRNA(s) reflects the epigenetic changes affecting the cellular processes in multiple disorders. To understand the complex effect of dysregulated microRNAs linked to neurodegeneration, we performed a cross-sectional microRNA expression analysis in idiopathic Parkinson's disease (n = 367), progressive supranuclear palsy (n = 35) and healthy controls (n = 416) from the Luxembourg Parkinson's Study, followed by prediction modelling, enriched pathway analysis and target simulation of dysregulated microRNAs using probabilistic Boolean modelling. Forty-six microRNAs were identified to be dysregulated in Parkinson's disease versus controls and 16 in progressive supranuclear palsy versus controls with 4 overlapping significantly dysregulated microRNAs between the comparisons. Predictive power of microRNA subsets (including up to 100 microRNAs) was modest for differentiating Parkinson's disease or progressive supranuclear palsy from controls (maximal cross-validated area under the receiver operating characteristic curve 0.76 and 0.86, respectively) and low for progressive supranuclear palsy versus Parkinson's disease (maximal cross-validated area under the receiver operating characteristic curve 0.63). The enriched pathway analysis revealed natural killer cell pathway to be dysregulated in both, Parkinson's disease and progressive supranuclear palsy versus controls, indicating that the immune system might play an important role in both diseases. Probabilistic Boolean modelling of pathway dynamics affected by dysregulated microRNAs in Parkinson's disease and progressive supranuclear palsy revealed partially overlapping dysregulation in activity of the transcription factor EB, endoplasmic reticulum stress signalling, calcium signalling pathway, dopaminergic transcription and peroxisome proliferator-activated receptor gamma coactivator-1α activity, though involving different mechanisms. These findings indicated a partially convergent (sub)cellular end-point dysfunction at multiple levels in Parkinson's disease and progressive supranuclear palsy, but with distinctive underlying molecular mechanisms.

In silico exploration of bioactive compounds from Acorus calamus L. for targeted treatment of ischemic heart disease: Molecular insights and therapeutic prospects

Abstract BACKGROUND: Ischemic heart disease (IHD) accounts for 80%–85% of mortality, highlighting the need to develop effective and noninvasive management strategies. Lekhana karma (therapeutic scrapping) helps in balancing the imbalanced Kapha and Medodhatu, which are important factors in the pathogenesis of IHD. Vacha—Acorus calamus Linn (AC) is 1 of the 10 Lekhaneeya drugs described in Charaka Samhita and is widely available and cost-effective. However, its bioactive phytoconstituents’ mode of action and molecular entities remain unexplored. Hence, this study aimed to elucidate the mode of action of its bioactive compounds and their interaction with IHD disease targets and pathways. METHODS: Phytochemicals from AC were collected, followed by screening of bioactive phytochemicals, collecting gene identifications, and pathways related to IHD. Later, common compounds and pathways were identified, and common targets between phytochemicals and pathways were sorted. A further network of phytochemicals-pathways-targets was constructed followed by molecular docking of highly enriched disease targets and phytochemicals. RESULTS: An in silico study revealed that 19 main active compounds of AC are interacting with 48 targets involved in regulating biological processes in the IHD. Molecular docking analyses were performed to explore the potential and affinity of these compounds with disease targets. Galangin, alpha-asarone, beta-asarone, and isoelemicin exhibited significant interactions with IHD disease targets, such as prostaglandin G/H synthase 2, cytochrome P450 1A2, transcription factor p65, vascular endothelial growth factor A, and tyrosine-protein kinase. Additionally, the study identified the interaction between the top 4 phytochemicals and the five most enriched disease targets. CONCLUSION: These findings provide a promising avenue for transforming bioactive phytoconstituents in AC into novel drug entities for treating IHD. The in silico approach identifies therapeutic targets, guides wet lab studies, and aids multi-compound management for IHD.

Abstract 3077: Identifying Regulators Of Human Adventitial Fibroblast Activation Through A Genome-wide CRISPR Screen

Background: Vascular diseases, including atherosclerosis, are the leading cause of death in the US and world-wide. A quiescent population of fibroblasts that reside in the adventitia (AdvFib) activates and expands upon different vascular insults and contribute to the progression of vascular diseases. However, what regulates AdvFib’s activation remains unknown. We designed a genome-wide CRISPRi screen on human AdvFib to identify these regulators. Methods: We generated an immortalized human coronary AdvFib line that stably expresses dCas9-KRAB, which effectively silences any genes of interest. These cells were infected with a pool of lentivirus carrying guide RNAs (gRNA) covering the entire transcriptome at 200X coverage and allowed to expand freely. Frequency of gRNAs was assessed at different cell passages to identify those that influence AdvFib activation and was followed by experimental validation. Lastly, bioinformatic analysis was performed to determine regulatory pathways and gene networks involved in AdvFib activation. Results: Comparing relative gRNA abundances from early to late passages revealed several hundred putative regulators of AdvFib activation. GO biological process analysis of the top 250 enriched gRNA targets demonstrated disproportionate representation of cyclin inhibitors, validating the effectiveness of our screening. Molecular signatures for adipogenesis and key regulators of tissue mineralization including IBSP and ENPP1 were notably enriched in our screen. Human disease association using PhenGenI revealed a significant association with atherosclerosis. Indeed, numerous coronary artery disease GWAS genes including TCF21 , ZEB2 , CDKN1A , and CDKN2A were enriched in our screen, providing additional support for AdvFib’s importance in atherosclerosis. These hits were subsequently validated individually in vitro. Conclusion: A genome-wide CRISPRi screen of AdvFib activation modulators identified numerous potential regulator genes, many of which have known genetic and observational association with human vascular diseases. This study affirms the potentially important role of AdvFib in human vascular disease and may pave the way to future therapeutics targeting this often neglected cell population.

Proton-induced reactions for 47Sc (and 46Sc) production: new nuclear cross section measurements on enriched titanium targets

47Sc is a theranostic radionuclide under the spotlight of the scientific community thanks to its potential for SPECT imaging and therapeutic applications. This work presents the recent measurements of proton-induced nuclear reaction cross-sections aimed at 47Sc production using enriched 48Ti, 49Ti and 50Ti targets from 23 up to 70 MeV. Since the co-production of contaminant isotopes is a key issue, and 46Sc is the main one having a longer half-life than 47Sc, the 48/49/50Ti(p,x)47Sc and 46Sc cross sections are presented and compared with the scarce literature data and TALYS estimations.