Experimental Proof Research Articles

Identification of two flavonoids antiviral inhibitors targeting 3C-like protease of porcine epidemic diarrhea virus.

Porcine epidemic diarrhea virus (PEDV) has caused severe damage to the global pig industry in the past 20 years, creating an urgent demand for the development of associated medications. Flavonoids have emerged as promising candidates for combating coronaviruses. It is believed that certain flavonoids can directly inhibit the 3C-like protease (3CLpro), thus displaying antiviral activity against coronaviruses. In this investigation, we applied a flavonoid library to screen for natural compounds against PEDV 3CLpro. Baicalein and baicalin were found to efficiently inhibit PEDV 3CLproin vitro, with the IC50 value of 9.50 ± 1.02 μM and 65.80 ± 6.57 μM, respectively. A docking analysis supported that baicalein and baicalin might bind to the active site and binding pocket of PEDV 3CLpro. Moreover, both baicalein and baicalin successfully suppressed PEDV replication in Vero and LLC-PK1 cells, as indicated by reductions in viral RNA, protein, and titer. Further investigation revealed that baicalein and baicalin mainly inhibited the early viral replication of the post-entry stage. Furthermore, baicalein showed potential effects on the attachment or invasion step of PEDV. Collectively, our findings provide experimental proof for the inhibitory effects of baicalein and baicalin on PEDV 3CLpro activity and PEDV infection. These discoveries may introduce novel therapeutic strategies for controlling porcine epidemic diarrhea (PED).

The host-targeted antiviral drug Zapnometinib exhibits a high barrier to the development of SARS-CoV-2 resistance

Host targeting antiviral drugs (HTA) are directed against cellular mechanisms which can be exploited by viruses. These mechanisms are essential for viral replication, because missing functions cannot be compensated by the virus. However, this assumption needs experimental proof. Here we compared the HTA Zapnometinib (ZMN), with direct acting antivirals (DAA) (Remdesivir (RDV), Molnupiravir (MPV), Nirmatrelvir (NTV), Ritonavir (RTV), Paxlovid PAX)), in terms of their potency to induce reduced drug susceptibilities in SARS-CoV-2. During serial passage of δ-B1.617.2 adaptation to all DAAs occurred, while the inhibitory capacity of ZMN was not altered. Known single nucleotide polymorphisms (SNPs) responsible for partial resistances were found for RDV, NTV and PAX. Additionally, the high mutagenic potential of MPV was confirmed and decreased drug efficacies were found for the first time. Reduced DAA efficacy did not alter the inhibitory potential of ZMN. These results show that ZMN confers a high barrier towards the development of viral resistance and has the potential to act against partially DAA-insensitive viruses.

Mechanical and Durability Studies about the use of Municipal Solid Waste Landfill in Concrete

The best way to keep the required format of the manuscript is to overwrite these instructions with its text. Papers should be written in English. Manuscripts s Landfilling is the most common and cheapest method of waste management practice in India. Municipal Solid Waste Landfills (MSWL) became a nuisance affecting the health, hygiene, sanitation and aesthetics of the surrounding area. Aggregates occupy almost 70% of concrete, so replacing waste materials with them could be a rewarding choice. In the current work, an experimental investigation is being carried out to test the addition of MSWL as a substitution with fine aggregate for concrete production. Out of the different aged samples available at the dumpsite, the most aged sample is chosen for experimental investigations according to the basic physical properties. Concrete mixes, with 0%, 4%, 5%, 7% and 10% partial replacement of fine aggregate with MSWL are tested for mechanical properties such as compressive strength, split tensile strength, flexural strength, and non-destructive test and have proved to be a partial substitute for fine aggregate. Durability studies such as water absorption, acid attack and sulphate attack also gave better experimental proof for the sustainable reuse of this waste material. The research reveals 5% replacement is the optimum considering all the test result values. The paper leads to advanced research for the suitability of the material in the construction industry.

Unveiling the experimental proof of the anticancer potential of ginsenoside Rg3 (Review).

Ginsenoside Rg3 (GS-Rg3), a sterol molecule isolated from ginseng, has demonstrated various immunological properties, including inhibition of cancer cell proliferation and metastasis, reversal of drug resistance and enhancement of chemotherapy sensitivity. The recent surge in attention towards GS-Rg3 can be attributed to its potential as an antitumor angiogenesis agent and as a therapeutic candidate for immunotherapy. The development of GS-Rg3 as an agent for these purposes has accelerated research on its mechanisms of action. The present review summarizes recent studies investigating the antitumor activity of GS-Rg3 and its underlying mechanisms, as well as providing essential information for future studies on GS-Rg3.

Toxicity of substituted p-phenylenediamine antioxidants and their derived novel quinones on aquatic bacterium: Acute effects and mechanistic insights

Substituted para-phenylenediamines (PPDs) are synthetic chemicals used globally for rubber antioxidation, with their quinone derivatives (PPD-Qs) raising particular environmental concerns due to their severe toxicity to aquatic organisms. Emerging research has identified a variety of novel PPD-Qs ubiquitously detected in the environment, yet experimental proof for the toxicity of PPD-Qs has not been forthcoming due to the unavailability of bulk standards, leaving substantial gaps in the prioritization and mechanistic investigation of such novel pollutants. Here, we use synthesized chemical standards to study the acute toxicity and underlying mechanism of 18 PPD-Qs and PPDs to the aquatic bacterium V. fischeri. Bioluminescence inhibition EC50 of PPD-Qs ranged from 1.76–15.6 mg/L, with several emerging PPD-Qs demonstrating significantly higher toxicity than the well-studied 6PPD-Q. This finding suggests a broad toxicological threat PPD-Qs pose to the aquatic bacterium, other than 6PPD-Q. Biological response assays revealed that PPD-Qs can reduce the esterase activity, cause cell membrane damage and intracellular oxidative stress. Molecular docking unveiled multiple interactions of PPD-Qs with the luciferase in V. fischeri, suggesting their potential functional impacts on proteins through competitive binding. Our results provided crucial toxicity benchmarks for PPD-Qs, prioritized these novel pollutants and shed light on the potential toxicological mechanisms.

Sequences at gene segment termini inclusive of untranslated regions and partial open reading frames play a critical role in mammalian orthoreovirus S gene packaging.

Mammalian orthoreovirus (MRV) is a prototypic member of the Spinareoviridae family and has ten double-stranded RNA segments. One copy of each segment must be faithfully packaged into the mature virion, and prior literature suggests that nucleotides (nts) at the terminal ends of each gene likely facilitate their packaging. However, little is known about the precise packaging sequences required or how the packaging process is coordinated. Using a novel approach, we have determined that 200 nts at each terminus, inclusive of untranslated regions (UTR) and parts of the open reading frame (ORF), are sufficient for packaging S gene segments (S1-S4) individually and together into replicating virus. Further, we mapped the minimal sequences required for packaging the S1 gene segment into a replicating virus to 25 5' nts and 50 3' nts. The S1 UTRs, while not sufficient, were necessary for efficient packaging, as mutations of the 5' or 3' UTRs led to a complete loss of virus recovery. Using a second novel assay, we determined that 50 5' nts and 50 3' nts of S1 are sufficient to package a non-viral gene segment into MRV. The 5' and 3' termini of the S1 gene are predicted to form a panhandle structure and specific mutations within the stem of the predicted panhandle region led to a significant decrease in viral recovery. Additionally, mutation of six nts that are conserved across the three major serotypes of MRV that are predicted to form an unpaired loop in the S1 3' UTR, led to a complete loss of viral recovery. Overall, our data provide strong experimental proof that MRV packaging signals lie at the terminal ends of the S gene segments and offer support that the sequence requirements for efficient packaging of the S1 segment include a predicted panhandle structure and specific sequences within an unpaired loop in the 3' UTR.

NiCoP fibers as novel catalysts for hydrogen evolution in alkali and acidic environment

NiCoP transition metal phosphide has been recognized as a most promising candidate to alternate the noble metals in hydrogen evolution reaction. Herein, it is first time presented the facile preparation of NiCoP electrocatalyst in form of fibers with dominant 1D structure formed by needle-less electrospinning technology and finalized by defined heat treatment process. The study provides a precise method for the preparation of NiCoP or similar phosphide fibers, together with an experimental proof of their excellent electrocatalytic performance in alkali and acidic environments. The as-prepared fibrous NiCoP electrocatalyst sintered in air at 900 °C followed by sintering at 700 °C in Ar/H2 exhibits low hydrogen evolution reaction overpotential (η10) of 141 mV in alkali and 146 mV in acidic environment corresponding with low Tafel slopes of 53 and 97.8 mV/dec, respectively. The bimetallic phosphide also shows superior oxygen evolution reaction activity in alkaline medium 1 M KOH with satisfying durability in long-term stability tests. The highly innovative fibrous form of the material not only plays a significant role in regulation of the electronic structure and boosting the electrochemical active sites and thus the electrocatalytic performance in water splitting technology, but also allows tuning of the electrode electric conductivity, occurrence of the active sites probability and thus utilisation of the electrode, together with its permeability and mechanical stability.

Unearthing a Cryptic Biosynthetic Gene Cluster for the Piperazic Acid-Bearing Depsipeptide Diperamycin in the Ant-Dweller Streptomyces sp. CS113.

Piperazic acid is a cyclic nonproteinogenic amino acid that contains a hydrazine N-N bond formed by a piperazate synthase (KtzT-like). This amino acid, found in bioactive natural products synthesized by non-ribosomal peptide synthetases (NRPSs), confers conformational constraint to peptides, an important feature for their biological activities. Genome mining of Streptomyces strains has been revealed as a strategy to identify biosynthetic gene clusters (BGCs) for potentially active compounds. Moreover, the isolation of new strains from underexplored habitats or associated with other organisms has allowed to uncover new BGCs for unknown compounds. The in-house "Carlos Sialer (CS)" strain collection consists of seventy-one Streptomyces strains isolated from the cuticle of leaf-cutting ants of the tribe Attini. Genomes from twelve of these strains have been sequenced and mined using bioinformatics tools, highlighting their potential to encode secondary metabolites. In this work, we have screened in silico those genomes, using KtzT as a hook to identify BGCs encoding piperazic acid-containing compounds. This resulted in uncovering the new BGC dpn in Streptomyces sp. CS113, which encodes the biosynthesis of the hybrid polyketide-depsipeptide diperamycin. Analysis of the diperamycin polyketide synthase (PKS) and NRPS reveals their functional similarity to those from the aurantimycin A biosynthetic pathway. Experimental proof linking the dpn BGC to its encoded compound was achieved by determining the growth conditions for the expression of the cluster and by inactivating the NRPS encoding gene dpnS2 and the piperazate synthase gene dpnZ. The identity of diperamycin was confirmed by High-Resolution Mass Spectrometry (HRMS) and Nuclear Magnetic Resonance (NMR) and by analysis of the domain composition of modules from the DpnP PKS and DpnS NRPS. The identification of the dpn BGC expands the number of BGCs that have been confirmed to encode the relatively scarcely represented BGCs for depsipeptides of the azinothricin family of compounds and will facilitate the generation of new-to-nature analogues by combinatorial biosynthesis.

Geometry-Inference Based Clustering Heuristic: New k-means Metric for Gaussian Data and Experimental Proof of Concept

Geometry-Inference Based Clustering Heuristic: New k-means Metric for Gaussian Data and Experimental Proof of Concept

Directly Identifying Multiple Cr3+ Emitting Centers for Broad Near‐Infrared Emission in an Efficient and Near‐Zero Thermal Quenching Garnet‐Type Phosphor

AbstractCr3+ doped garnet‐type (A3B2C3O12) near‐infrared (NIR) phosphors is a desirable research hotspot due to their prospective night vision, bioimaging, and plant growth utilizations. Meanwhile, the origin of multisite broad emission is yet a controversial topic. To resolve the trouble, verifying accurately the multiple Cr3+ occupancy sites is a key. Furthermore, it is an urgent need for improving the external quantum efficiency and luminescent thermostability. Herein, in Y3Sc2Al3O12 (YSAO), advanced electron microscopy technique is employed to directly confirm that the obtained broadband NIR emission originates from Cr3+ in [ScO6] octahedral and [YO8] dodecahedral sites. The optimal YSAO:5%Cr3+ sample exhibits high quantum efficiency (IQE/EQE = 74/31%) and near‐zero thermal quenching (97%@423 K and 92%@473 K). The theoretical calculations and experimental proofs reveal that YSAO possesses high structural rigidity and wide bandgap, which is responsible for the extremely thermostable luminescence. The high‐power YSAO:5%Cr3+‐converted NIR LED device shows promising multifunctional applications. This work not only provides an efficient NIR broadband phosphor with near‐zero thermal quenching, but also provides an effective determination method for the multiple emitting centers of phosphor materials.

Metabolites of Clostridium leptum fermenting flaxseed polysaccharide alleviate obesity in rats

Obesity is a chronic metabolic disease. Our previous research found flaxseed polysaccharide (FP) has an anti-obesity effect, and its anti-obesity effect possibly depends on Clostridium leptum (C. leptum). However, whether the strain takes the role and how it works is still being determined. Here, FP was fermented in vitro by C. leptum and its metabolites were analyzed. Subsequently, the FP fermentation broth of C. leptum (FPF) was given to the obese pseudo sterile rats. The results showed FPF was rich in various metabolites, among which the top ten in relative expression abundance were 3 beta-hydroxy-5-cholestenoate, 7,8-dihydro-3b,6a-dihydroxy-alpha-ionol 9-glucoside, Valyl-Serine, 2-amino-4-[(2-hydroxy-1-oxopropyl)amino]butanoic acid, Agavoside B, glycylproline, lycopersiconolide, armillaritin, Isoleucyl-Hydroxyproline and norethindrone acetate. After intervention with FPF, the weight, abdominal fat ratio, and total fat ratio of rats were significantly reduced and the lipid metabolism of them has been improved. This effect may be achieved by up regulating glucagon-like peptide-1 and adiponectin and further activating the AMP-activated protein kinase signaling pathway. This is the first experimental proof that FP exerts its anti-obesity effects through metabolites from C. leptum fermenting FP, not FP itself and the bacterial cells (debris) of C. leptum. It is also the first demonstration that FPF has a significant anti-obesity effect.

Time-resolved photoelectron spectroscopy of iodide-4-thiouracil cluster: The ππ* state as a doorway for electron attachment.

The photophysics of thiobases-nucleobases in which one or more oxygen atoms are replaced with sulfur atoms- vary greatly depending on the location of sulfonation. Not only are direct dynamics of a neutral thiobase impacted, but also the dynamics of excess electron accommodation. In this work, time-resolved photoelectron spectroscopy is used to measure binary anionic clusters of iodide and 4-thiouracil, I- · 4TU. We investigate charge transfer dynamics driven by excitation at 3.88eV, corresponding to the lowest ππ* transition of the thiouracil, and at 4.16eV, near the cluster vertical detachment energy. The photoexcited state dynamics are probed by photodetachment with 1.55 and 3.14eV pulses. Excitation at 3.88eV leads to a signal from a valence-bound ion only, indicating a charge accommodation mechanism that does not involve a dipole-bound anion as an intermediate. Excitation at 4.16eV rapidly gives rise to dipole-bound and valence-bound ion signals, with a second rise in the valence-bound signal corresponding to the decay of the dipole-bound signal. The dynamics associated with the low energy ππ* excitation of 4-thiouracil provide a clear experimental proof for the importance of localized excitation and electron backfilling in halide-nucleobase clusters.

First experimental demonstration of real-time neutron capture discrimination in helium and carbon ion therapy

This work provides the first experimental proof of an increased neutron capture photon signal following the introduction of boron to a PMMA phantom during helium and carbon ion therapies in Neutron Capture Enhanced Particle Therapy (NCEPT). NCEPT leverages 10\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$^{10}$$\\end{document}B neutron capture, leading to the emission of detectable 478 keV photons. Experiments were performed at the Heavy Ion Medical Accelerator in Chiba, Japan, with two Poly(methyl methacrylate) (PMMA) targets, one bearing a boron insert. The BeNEdiCTE gamma-ray detector measured an increase in the 478 keV signal of 45 ± 7% and 26 ± 2% for carbon and helium ion irradiation, respectively. Our Geant4 Monte Carlo simulation model, developed to investigate photon origins, found less than 30% of detected photons originated from the insert, while boron in the detector’s circuit boards contributed over 65%. Further, the model investigated detector sensitivity, establishing its capability to record a 10% increase in 478 keV photon detection at a target 10\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$^{10}$$\\end{document}B concentration of 500 ppm using spectral windowing alone, and 25% when combined with temporal windowing. The linear response extended to concentrations up to 20,000 ppm. The increase in the signal in all evaluated cases confirm the potential of the proposed detector design for neutron capture quantification in NCEPT.

Labile silver(I) complexes with benzisothiazolinone anion and phosphines: Synthesis, characterization and hydrogen storage application

AbstractA silver complex was prepared with benzisothiazolinone ligand, which was used to prepare new phosphorus‐rich complexes with dppe and dppp. All complexes were diagnosed by FTIR, 1H‐NMR, 31P‐NMR, CHN, and molar conductivity. The results proved that phosphines were labile, therefore 31P‐NMR was measured by cryoprotection at different temperatures. As a promoted effort to obtain direct experimental proof for the dimeric and the labile nature of [Ag(bit)(diphos)]2 complexes in solution, the 31P{1H}‐NMR spectrum of mixed (dppe/dppp) ligand complexes was measured. It was anticipated that an exchange of phosphines occurred between [Ag(bit)(dppe)]2 and [Ag(bit)(dppp)]2 complexes. However, a distribution of the dppp and dppe should result, and complex with compositions of [Ag2(bit)2(dppe)(dppp)] will be formed. The phosphine‐containing complexes were used in hydrogen storage study. The results proved that hydrogen storage values were initially similar for [Ag(bit)(dppp)]2 and [Ag(bit)(dppe)]2, but increased significantly for [Ag(bit)(dppe)]2 at higher pressures to reach 4.35 wt% at 90 bar.

Functional determination of site-mutations in rdxA involved in metronidazole resistance of Helicobacter pylori.

Metronidazole (MTZ) is among the first-line drugs against the human gastric pathogen Helicobacter pylori (H. pylori). MTZ is used as a prodrug that is activated by an oxygen-insensitive enzyme NADPH nitroreductase (RdxA). Loss-of-function mutations in rdxA make H. pylori MTZ resistant; however, experimental proof is lacking. We collected 139 gastric biopsy samples from patients suspected of H. pylori infection in Shanghai, and amplified Hp-specific rdxA gene from 134 samples. All these rdxA genes were sequenced and phylogenetically compared. The effect of mutations on RdxA function was measured by expressing them in Escherichia coli DH5α by using the MTZ sensitivity test. In total, 134 gastric biopsy samples were identified as H. pylori positive. Of the 134 samples, 74 and 6 had point mutations at the various sites or promoter region of rdxA, generating truncated and extended fused proteins, respectively. The remaining 54 were full-length with single nucleotide variation (SNV) compared with the wild-type RdxA from H. pylori, with 49 clustering with hpEastAsia, 3 with hpEurope, and 2 with hpNEAfrica. All 134 rdxA were expressed in E. coli DH5α; 22 and 112 resultant strains showed MTZ-sensitive and MTZ-resistant phenotypes, respectively. Comparative analysis of single nucleotide polymorphisms (SNPs) in the functional and inactivated RdxA revealed 14 novel mutations in RdxA, 5 of which conferred MTZ resistance: S18F, D59S, L62I, S79N, and A187V. The occurrence of MTZ resistance induced by site-mutation of RdxA in patients with H. pylori infection was 83.6% (112/134) in the Shanghai region. The major form of loss-of-function mutation was truncation of RdxA translation at a rate of 58/112 (51.8%). Molecular detection reliably determined the resistance of H. pylori to MTZ. Thus, the functional mutants involved in MTZ resistance facilitate clinical diagnosis and medication based on sequence analysis.

Observation of piezoelectricity in a lead-free Cs2AgBiBr6 perovskite: a new entrant in the energy harvesting arena.

Halide perovskite materials have recently been recognised as powerful ferroelectric and piezoelectric materials with applications in the energy harvesting arena, but their experimental proof is very limited. We achieved strong intrinsic piezoelectricity in the lead-free inorganic double perovskite Cs2AgBiBr6 at room temperature and utilized it for mechanical energy harvesting, with a piezoelectric co-efficient (d33) of 12.7 pC N-1. Hysteresis loop and structural analyses offered further validation for the substantial ferroelectric features of the as-synthesised double perovskite. Density functional theory (DFT) calculations revealed the presence of anharmonic phonon soft modes in tetragonal Cs2AgBiBr6 due to dynamic instability, which resulted in piezoelectricity. Under an optimal pressure of ≈25 kPa, a Cs2AgBiBr6 thin film-based piezoelectric nanogenerator device delivered instantaneous output values of ≈45 V and ≈200 nA. The strain-sensitive responses of the device were also exemplified to identify specific body motions from the detected instantaneous output values. The energy obtained from the device is shown to be effective for capacitor charging and commercial light-emitting diode (LED) lighting. Our study provides significant insights into the dielectric behaviour of materials as well as piezo- and ferroelectric behaviours, which are crucial for the development of modern electronic and energy harvesting devices.

Quercetin-induced pyroptosis in colon cancer through NEK7-mediated NLRP3 inflammasome-GSDMD signaling pathway activation.

Pyroptosis, a gasdermin-mediated lytic cell death, is a new hotspot topic in cancer research, and induction of tumor pyroptosis has emerged as a new target in cancer management. Quercetin (Que), a natural substance, demonstrates promising anticancer action. However, further information is required to fully comprehend the function and mechanism of Que in pyroptosis in colon cancer. This study revealed the underlying mechanism of Que-induced pyroptosis in colon cancer in vitro and in vivo. Que inhibited colon cancer cell growth through gasdermin D (GSDMD)-mediated pyroptosis. Depletion of GSDMD, rather than gasdermin E (GSDME), reversed the cytotoxic effects of Que on colon cancer cells. Que treatment upregulated NIMA-related kinase 7 (NEK7) protein expression, thus facilitating the assembly of the NLRP3 inflammasome and cleavage of GSDMD. NEK7 silencing resulted in colon cancer cell growth in vitro and in vivo. Mechanistically, NEK7 depression restrained the activation of the NLRP3 inflammasome-GSDMD pathway, thus attenuating pyroptosis triggered by Que in colon cancer cells. Furthermore, lower NEK7 and NLRP3 expression levels indicated colon cancer progression. Our results unveiled a novel pattern of anti-colon cancer activity of Que, and activation of NEK7-mediated pyroptosis is potentially a promising therapeutic target for colon cancer, which provides novel experimental proof for the clinical application of Que.

Demonstration of a novel majority logic in a memristive crossbar array for in-memory parallel computing.

A memristive crossbar array can execute Boolean logic operations directly within the memory, which is highly noteworthy as it addresses the data bottleneck issue in traditional von Neumann computing. Although its potential has been widely demonstrated, achieving practical levels of operational reliability and computational efficiency remains a challenge. Here, we introduce a three-input majority logic gate supported by near-memory operations, serving as a universal gate and achieving both robust reliability and high efficiency in versatile logic operations. We fabricated a highly reliable HfOx-based memristive array, incorporating a series resistor to increase the reset voltage of the memristor, thereby increasing the operational voltage margin of the gate operation. This ensured reliable operation of the majority gate, resulting in successful experimental proof of combined 1-bit full adder and subtractor operations performed in 5 steps using 7 cells. Additionally, we propose that an N-bit parallel prefix adder (PPA) operation is possible in O(log2 N) steps, by taking advantage of the parallel operation capability of the majority gate. This achieves 8.5× higher spatiotemporal efficiency than the previously reported NOR-based logic system in 64-bit adder operation. Moreover, as N increases, the spatiotemporal efficiency further improves, which significantly enhances the applicability of memristive logic-in-memory.

Ex vivo platform en route to functional precision medicine: clinical relevance in gynecological cancers.

A scientific interrogation-driven approach to the clinical management of cancer patients is based on molecular profiling of the tumor. Empowered by the knowledge of oncogenic drivers and biomarkers, oncologists chart an optimal treatment path toward increasing the mathematical probability of a positive outcome. In this entire chain of events, an experimental proof of logical interrogation has never been incorporated before. Here, we provide the first evidence that the result of ex vivo testing of a drug matched to the genomic profiling of an N-of-1 tumor can deliver meaningful insight connecting scientific interrogation and a clinical event. Using resected tissues from endometrial (EC) and ovarian (OC) cancer patients, we designed a personalized ex vivo platform to test combinations of drugs in the default histological architecture of the individual tumors. Following the CART-T cells' principle, we co-cultured with autologous T-cells to test targeted drugs and immune checkpoint inhibitors. The study was designed with a limited clinical information window from patient registration/consent to obtaining the tumor tissues, and adjuvant treatment/post-surgery event (PSE) data were accessed retrospectively. Using a checkerboard analysis, we found that PSE-free survival time was longer in patients whose therapy "matched" the effective drug combination in ex vivo culture/co-cultures compared to those with no effect. Specifically, out of 32 EC patients in the "test & treatment-matched" category whose tumor cells failed to respond to ex vivo drug testing, none achieved > 4 and > 3 years of PSE-free survival. In contrast, out of 38 EC patients in the "test & treatment-matched" category, 4 and 6 patients, whose tumor cells responded to drugs in ex vivo culture, achieved > 4 and > 3 years of PSE-free survival, respectively. Cases with genomically-guided ex vivo testing showed that a "match" between an effective ex vivo drug combination and therapy resulted in late PSE, whereas a "match" between prescribed treatment and an ineffective drug combination in ex vivo testing led to early PSE. Our study demonstrates that integrating genomic data with personalized drug testing on an ex vivo culture/co-culture platform is an effective tool for modeling functional precision medicine in gynecological cancers. This approach bridges the gap between next-generation drug testing in translational research and patient care, providing insight for improved treatment outcomes.

Towards an experimental proof of the magnonic Aharonov-Casher effect

Towards an experimental proof of the magnonic Aharonov-Casher effect