Significant Activity Research Articles

TGF-β Promotes Hepatocellular Carcinoma Metastasis Through Through m6A Modification of PCDHGA9.

Objective: Hepatocellular carcinoma (HCC) is a highly lethal cancer with significant mortality, primarily attributed to metastasis. Although Protocadherin Gamma Subfamily A, 9 (PCDHGA9) has been identified as a tumor suppressor gene in cancer metastasis, its role in HCC remains ambiguous. This study aims to clarify the role of PCDHGA9 in HCC by examining its expression, clinical significance, and molecular activities. Methods: Tissue microarray immunofluorescence analysis evaluated the expression of PCDHGA9 and its clinical relevance. In vitro experiments involved manipulating PCDHGA9 levels in SK-HEP-1 cells to assess migration through wound-healing and transwell assays. In vivo, shPCDHGA9 cell injections were utilized to observe effects on tumor growth and metastasis. Protein analysis and Western Blot validated epithelial-mesenchymal transition (EMT)-related proteins. Subsequent to TGF-β treatment, cell proliferation and apoptosis were quantified using Cell counting kit-8 and flow cytometry, respectively, followed by investigation of TGF-β effects on PCDHGA9 N6-methyladenosine (m6A) modification via Methylated RNA immunoprecipitation, RT-qPCR, and Western blot analysis. Results: Downregulation of PCDHGA9 expression in HCC tissues is correlated with poor prognosis. In vitro experiments demonstrated that modulating PCDHGA9 expression influenced HCC cell migration. In vivo, PCDHGA9 knockdown is correlated with increased metastasis. Furthermore, TGF-β stimulation promoted cell proliferation and inhibited apoptosis. Mechanistically, TGF-β-mediated m6A modification led to PCDHGA9 decay, promoting EMT in HCC cells. Conclusion: PCDHGA9 serves as a potential tumor suppressor in HCC by inhibiting EMT. During this process, TGF-β is observed to exert regulatory control over m6A modifications of PCDHGA9.

Assessment of possible biomedical applications of green synthesized TiO2NPs-an in-vitro approach

Biomedical applications for various types of nanoparticles are emerging on a daily basis. Hence this research was performed to evaluate the antifungal (Aspergillus sp., Alternaria sp., Trichophyton sp., Candida sp., and Penicillium sp.), cytotoxicity (MCF10A cell lines), and antioxidant (DPPH) potential of Coleus aromaticus mediated and pre-characterized TiO2NPs were studied with respective standard methodology. Interestingly, the TiO2NPs exhibited significant antifungal activity on pathogenic fungal strains like Alternaria sp., Aspergillus sp. (31 ± 1.4), Penicillium sp. (31 ± 1.9) Trichophyton sp. (27 ± 2.1), and Candida sp. (26 ± 2.3) at high concentration (250 μg mL−1). However, the considerable levels of zone of inhibitions on fungal pathogens were recorded at 100 μg mL−1 of TiO2NPs as well as it was considerably greater than positive control. It also demonstrated dose based anti-inflammatory and antidiabetic activities. The plant-mediated TiO2NPs demonstrated a maximum DPPH scavenging efficiency of 91% at a dosage of 250 μg mL−1, comparable to the positive control's 94%. Furthermore, TiO2NPs at 100 μg mL−1 concentration did not cause cytotoxicity in MCF10A cell lines. At higher concentrations (250 μg mL−1), the nanoparticles showed the lowest cytotoxicity (17%). These findings suggest that C. aromaticus-mediated TiO2NPs have significant biomedical applications. However, in-vivo studies are needed to learn more about their (C. aromaticus-mediated TiO2NPs) potential biomedical applications.

Development and Evaluation of a Novel Hyaluronic Acid and Chitosan-modified Phytosome for Co-delivery of Oxymatrine and Glycyrrhizin for Combination Therapy.

Multidrug resistance (MDR) of cancer cells is a major obstacle to efficient cancer chemotherapy. Combination therapy is expected to enhance the anticancer effect and reverse MDR. Numerous patents involve different kinds of nanoparticles for the co-delivery of multiple chemotherapeutics, but the FDA has approved none. In this study, oxymatrine (OMT) and glycyrrhizin (GL) were co-loaded into phytosomes as the core of nanocarriers, and the shell was cross-linked with chitosan (CS) and hyaluronic acid (HA) with the capability for the controlled, sequential release and the targeted drug uptake. Phospholipid complexes of OMT and GL (OGPs) were prepared by a solvent evaporation technique and could self-assemble in an aqueous solution to form phytosomes. CS and HA were sequentially coated on the surface of OGPs via electrostatic interactions to obtain CS coated OGPs (CS-OGPs) and HA modified CS-OGPs (HA-CS-OGPs), respectively. The particle size and zeta potential were measured to optimize the formulations. In vitro cytotoxicity and cellular uptake experiments on HepG2 cells were performed to evaluate the anticancer activity. OGPs were obtained with nano-size around 100 nm, and CS and HA coating on phytosomes could change the particle size and surface potential. The drug loading of OMT and GL showed that the nanocarriers could maintain a fixed ratio of 1:1. The in vitro release experiments indicated the release of OMT and GL was pH-dependent and sequential: the release of OMT from CS-OGPs and HA-CS-OGPs was significantly increased at pH 5.0 compared to the release at pH 7.4, while GL exhibited sustained released from CS-OGPs and HA-CS-OGPs at pH 5.0. Furthermore, in vitro cytotoxicity and cellular uptake experiments on HepG2 cells demonstrated that the co-delivery system based on phytosomes had significant synergistic anti-tumor activities, and the effects were enhanced by CS and HA modification. The delivery of OMT and GL via HA-CS-OGPs might be a promising treatment to reverse MDR in cancer therapy.

Effects of 90 dB pure tone exposure on auditory and cardio-cerebral system functions in macaque monkeys

Excessive noise exposure can pose significant health risks to humans. In addition to causing direct damage to the auditory system, noise exposure is also associated with pathological changes or injuries in the cardiovascular and central nervous systems. Uncovering the physiological effects of noise exposure on the body contributes to a better understanding of the potential health risks associated with noise exposure, aiding in the establishment and updating of safety standards and limits for occupational noise exposure. This study focused on three male macaque monkeys as subjects. Low-intensity pure tone exposure (intensity: 90 dB, frequency: 500 Hz, repetition rate: 40 Hz, 10 min per day, continuously exposed for 5 days) was administered. Evaluations were conducted before noise exposure, immediately after exposure, and at 7, 14, and 28 days post-exposure. Auditory brainstem response (ABR) tests were conducted to assess the impact of noise exposure on the hearing of the macaque monkeys. Electrocardiograms (ECG) were performed to evaluate changes in the heart function of the monkeys after noise exposure. Electroencephalograms (EEG) were recorded to assess noise-induced changes in brain function. The results of the study revealed that, compared to pre-exposure levels, the average threshold for the V wave of the right ear of the Macaque monkeys increased by approximately 30 dB immediately after exposure (p < 0.01), and it returned to normal within 7 days. The ECG results indicated that one of the macaque monkeys exhibited an RS-type QRS wave, and inverted T waves from immediately after exposure to 14 days, which normalized at 28 days. The other two macaque monkeys showed no significant changes in their ECG parameters. Changes in EEG parameters demonstrated that main brain regions exhibited significant activation at 40 Hz during noise exposure. Noise stimulation with a 40 Hz repetition rate induced abnormal resting-state electroencephalogram power spectral density (PSD) across the δ (1–3 Hz), θ (4–8 Hz), α (8–12 Hz), β1 (12–20 Hz), β2 (20–30 Hz), and γ (30–48 Hz) frequency bands. After noise exposure, the PSD in main brain regions, particularly those represented by the temporal lobe, exhibited a decreasing trend across all frequency bands, with no clear recovery over time. In summary, exposure to 90 dB pure tone resulted in transient hearing threshold shifts, inverted T waves, activation of brain regions at 40 Hz, and disrupted PSD of resting-state EEG in macaque monkeys. While these effects were observed, they exhibited individual differences. This study investigated the physiological changes in auditory and cardio-cerebral systems in macaque monkeys following low-intensity noise exposure, which could be informative for noise protection and the establishment of safety standards for human. However, the exact mechanisms involved require further pathological and behavioral exploration.

Examining contrasting influences of extreme weather experiences on individual climate activism

Researchers have examined how extreme weather experiences influence climate change attitudes, beliefs, and behaviors, with mixed results. However, limited research has explored how extreme weather experiences may affect climate-related perceptions and behaviors among climate activists. Given the significant role activism plays in climate action, as well as frequent dropout and burnout among activists, it is important to understand to what extent, how, and why extreme weather may influence individual climate activism. This study explores reported influences of extreme weather experiences on climate perceptions and activism through interviews with 33 Australian adults who directly experienced bushfires and previously engaged in climate activism. All participants felt more vulnerable to climate change after experiencing bushfires. Fifteen participants (45%) increased their activism; 13 (39%) maintained the same activism level; and 5 (15%) decreased their activism. Participants who increased their activism sought to share their bushfire stories with news media, policymakers, and through artistic projects. Climate activism helped several participants cope with bushfire-related trauma, whereas several other participants reduced their activism because their experiences undermined self-efficacy (perception that one can act on climate change). These findings show the divergent ways individuals may respond to extreme weather experiences and have implications for climate action mobilization strategies.

A Snapshot of Selenium-enclosed Nanoparticles for the Management of Cancer.

Among the primary causes of mortality in today's world is cancer. Many drugs are employed to give lengthy and severe chemotherapy and radiation therapy, like nitrosoureas (Cisplatin, Oxaliplatin), Antimetabolites (5-fluorouracil, Methotrexate), Topoisomerase inhibitors (Etoposide), Mitotic inhibitors (Doxorubicin); such treatment is associated with significant adverse effects. Antitumor antibiotics have side effects similar to chemotherapy and radiotherapy. Selenium (Se) is an essential trace element for humans and animals, and additional Se supplementation is required, particularly for individuals deficient in Se. Due to its unique features and high bioactivities, selenium nanoparticles (SeNPs), which act as a supplement to counter Se deficiency, have recently gained worldwide attention. This study presented a safer and more economical way of preparing stable SeNPs. The researcher has assessed the antiproliferative efficiency of SeNPs-based paclitaxel delivery systems against tumor cells in vitro with relevant mechanistic visualization. SeNPs stabilized by Pluronic F-127 were synthesized and studied. The significant properties and biological activities of PTX-loaded SeNPs on cancer cells from the lungs, breasts, cervical, and colons. In one study, SeNPs were formulated using chitosan (CTS) polymer and then incorporated into CTS/citrate gel, resulting in a SeNPs-loaded chitosan/citrate complex; in another study, CTS was used in the synthesis of SeNPs and then situated into CTS/citrate gel, resulting in Se loaded nanoparticles. These formulations were found to be more successful in cancer treatment.

Fluoroquinolone and enoxacin molecules are potential urease inhibitors for treating ureolytic bacterial infections

Urease, a nickel-dependent metalloenzyme, has emerged as a significant therapeutic target due to its role in promoting the pathogenesis of various human health conditions. These include the development of pyelonephritis, urolithiasis, peptic ulcers, hepatic encephalopathy, and gastric ulcers. The currently available treatment involved the usage of strong antibiotics along with proton pump inhibitors to cope with the infection of urease producing bacteria. These conventional treatments are becoming less effective as bacteria are gaining multiple drug resistance. Therefore, there is a crucial need to identify alternative compounds with potential anti-urease activity and minimal side effects. Fluoroquinolones and Enoxacin derivatives offer antimicrobial, anti-inflammatory, antioxidant, anti-diabetic, and anti-urease activities. To improve the chemical diversity of urease inhibitors, different series of fluoroquinolones and Enoxacin derivatives were evaluated against urease and their antioxidant activity was also evaluated. To achieve this objective, in-silico studies were conducted utilizing molecular docking and adsorption, distribution, metabolism, excretion, and toxicology (ADMET) models. These analyses were employed to explore potential binding mechanisms and assess the drug-likeness of the compounds against urease enzymes. The inhibitory effect of docked heterocyclic compounds was also verified in-vitro against urease enzyme. Fluoroquinolones derivatives were found to be active inhibitors at high dose levels but showed minimum inhibition at low concentration. The compound EN from the Enoxacin series exhibited the highest potency as a urease inhibitor, with an IC50 of 45.86 μM, out-performing the standard drug thiourea, which had an IC50 of 52.20 μM. Additionally, compounds NOX-3 and FB-17 from the fluoroquinolone and Enoxacin series demonstrated significant DPPH free radical scavenging activity, with IC50 values of 98.17 μM and 97.98 μM, respectively. These results were comparable to the positive control ascorbic acid, which had an IC50 of 48.15 μM. This study demonstrates that Enoxacin derivatives can be further analyzed as potent urease inhibitors, while both Enoxacin and fluoroquinolone derivatives can be developed into more effective drugs to overcome oxidative stress.

Antimicrobial and cytotoxic naphthoquinones from microbial origin: An updated review.

Naphthoquinones (NQs) are small molecules bearing two carbonyl groups. They have been the subject of much research due to their significant biological activities such as antiproliferative, antimicrobial, anti-inflammatory, antioxidant, and antimalarial effects. NQs are produced mainly by bacteria, fungi and higher plants. Among them, microorganisms are a treasure of NQs with diverse skeletons and pharmacological properties. The purpose of the present study is to provide a comprehensive update on the structural diversity and biological activities of 91 microbial naphthoquinones isolated from 2015 to 2022, with a special focus on antimicrobial and cytotoxic activities. During this period, potent cytotoxic NQs such as naphthablin B (46) and hygrocin C (30) against HeLa (IC50=0.23 µg/ml) and MDA-MB-431 (IC50=0.5 µg/ml) cell lines was reported, respectively. In addition, rubromycin CA1 (39), exhibited strong antibacterial activity against Staphylococcus aureus (MIC of 0.2 µg/ml). As importance bioactive compounds, NQs may open new horizon for treatment of cancer and drug resistant bacteria. As such, it is hoped that this review article may stimulates further research into the isolation of further NQs from microbial, and other sources as well as the screening of such compounds for biological activity and beneficial uses.

Design, Synthesis and Anti-cancer Evaluation of Nitrogen-containing Derivatives of 30-Carboxyl of Gambogic Acid.

Gambogic acid (GA) is a natural product from the resin of the Garcinia species, which showed significant activity in the induction of apoptosis. It can be one promising lead compound for the design and synthesis of new anticancer drugs. The objective of the current study is to design novel nitrogen-contained GA derivatives with better anti-cancer activities and study the effect of the introduction of different nitrogen-contained groups on the activity of GA. The designed 15 derivatives were synthesized via esterification or amidation of 30-carboxylate. The synthetic compounds were characterized via different spectroscopic techniques, including X-ray single crystal diffraction, MS and NMR. The cytotoxic activity of the designed derivatives was evaluated in vitro against A549, HepG-2, and MCF-7 cell lines using methyl thiazolyl tetrazolium (MTT) test. 15 nitrogen-contained GA derivatives were successfully synthesized and established. Based on the IC50 values, compounds 9, 10, 11 and 13 showed stronger inhibitory effects on A549, HepG-2, MCF-7 cell lines than GA, while 9 is the most active compound with IC50 value of 0.64-1.49 μM. Most derivatives of GA with esterification of C-30 including cyano-benzene ring were generally weaker than those of pyrimidinyl-substituted derivatives. In addition, length of alkyl linkers between C-30 of GA and nitrogen-contained group produced different effects on A549, HepG-2 and MCF-7 cell lines. The structure-activity relationship results show that aromatic substituent and linker length play important roles to improve the anticancer activities, while compound 9 with pyrimidine substituent and C-C-C linkers is the most active derivative against tested cell lines, and is a promising anti-cancer agent for further development.

Simultaneous effect of medicinal plants as natural photosensitizers and low-level laser on photodynamic inactivation.

Photodynamic inactivation (PDI) technology is a promising alternative to antibiotics. This technology is defined as the inhibition of bacterial growth with photosensitizers while irradiated with low-level laser light in the wavelength of 532 ± 2.08nm. A challenging area in this field is selecting photosensitizers with antibacterial potential. In this paper, to enhance the antibacterial efficiency, the photosensitizers (the selected plant extracts) with a high absorption peak at the selected laser frequency, 532nm, were prepared. Low-concentration ethanolic plant extracts of Hibiscus sabdariffa and Opuntia ficus-indica were found to exhibit significant antibacterial activity against, Acinetobacter baumannii ATCC 19606 and, Staphylococcus aureus ATCC 33591 as two important human pathogenic bacteria. The effectiveness of these natural photosensitizers was measured by determining their Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values and by performing a time-killing assay in the absence and the presence of laser irradiation. Our results showed that the combination of low-level laser irradiation and the selected photosensitizers had excellent potential for treating in vitro bacterial infections. Therefore, PDI technology has great potential as a viable alternative to traditional antibiotics for combating bacterial infections. This study presents a promising avenue for further exploration of PDI and the use of laser technology in medical science.

Effect of vitamin D3 supplementation on blood parameters and liver gene expression in female rats.

To better understand the molecular mechanism responsible for the therapeutic potential of vitamin D, we conducted an analysis of the liver transcriptomes of adult female rats. Adult female rats (n = 18) were divided into three groups, receiving different doses of vitamin D: group I, 0; group II, 1000 U/kg; and group III, 5000 U/kg. Growth, body weight, the weight of main organs, blood haematological and biochemical parameters were evaluated. Gene expression in the liver were analyzed using RNA-seq and qPCR techniques. We observed a lower platelet count (p < 0,008) and a significantly greater (p < 0.02) number of WBCs in rats supplemented with 1000 U/kg than in rats from group III (5000 U/kg). Moreover, we noted a trend (p < 0.06) in total cholesterol concentration, suggesting a linear decrease with increasing doses of vitamin D. RNA-seq analysis did not reveal any differentially expressed genes with FDR < 0.05. However, GSEA revealed significant activation of a number of processes and pathways, including: "metallothionein, and TspO/MBR family", and "negative regulation of tumor necrosis factor production". qPCR analysis revealed significant upregulation of the Mt1, Mt2 and Orm1 genes in animals receiving high doses of vitamin D (p < 0.025, p < 0.025, and p < 0009, respectively). Moreover, Srebp2 and Insig2 were significantly lower in both experimental groups than in the control group (p < 0.003 and p < 0.036, respectively). Our results support the anti-inflammatory, anitioxidant and anticholesterologenic potential of vitamin D but suggest that high doses of vitamin D are needed to obtain significant results in this regard.

Abstract 6487: The selective WEE1 inhibitor azenosertib shows synergistic anti-tumor activity with KRASG12C inhibitors in multiple KRASG12C models

Abstract KRASG12C is a potent oncogenic driver which results in downstream hyperactivation of MAPK signaling and unchecked oncogenic growth, while simultaneously increasing replication stress (RS) and accumulation of DNA damage. KRASG12C-specific inhibitors are approved and/or recommended in metastatic non-small cell lung cancer (NSCLC) and colorectal cancer (CRC), and while responses are robust, some patients are inherently resistant, and most responders acquire resistance. Resistance mechanisms include reactivation of MAPK signaling, and cancer cells which escape MAPKi are reported to have high levels of genomic instability. WEE1 is a kinase involved in cell cycle progression, allowing cells to repair damaged DNA before entering the next cell cycle phase. Consequently, inhibition of WEE1 leads to premature cell cycle entry and RS, thereby driving cancer cells through the cell cycle with unchecked accumulation of DNA damage, ultimately resulting in mitotic catastrophe and cell death. We hypothesized that high levels of genomic instability and RS in KRASG12C mutant and KRASG12C inhibitor-resistant tumors present a vulnerability to WEE1 inhibition and provide rationale for combining azenosertib, a novel, selective, and orally bioavailable WEE1 inhibitor, with KRASG12C inhibitors. Here, we report azenosertib in combination with KRASG12C inhibitors as an effective therapeutic strategy for KRASG12C tumors. In vitro combination of azenosertib with multiple KRASG12C inhibitors demonstrated synergistic cell growth inhibition across a panel of KRASG12C cell lines in both 2D and 3D assays. In vivo treatment with azenosertib in KRASG12C inhibitor-sensitive cell line-derived xenograft (CDX) models of NSCLC (NCI-H358, NCI-H2122, NCI-H1792), CRC (SW837, SW1463), and pancreatic cancer (MIA PaCa-2) demonstrated significant monotherapy activity as well as synergistic tumor growth inhibition (TGI) when combined with KRASG12C inhibitors, including tumor regression in some models. Importantly, the KRASG12C inhibitor-resistant SW1573 NSCLC CDX model demonstrated synergistic TGI in the combo arm (84%) when compared against monotherapy azenosertib (31%) or adagrasib (58%). Similarly, treatment of a KRASG12C inhibitor-resistant patient-derived xenograft (PDX) model of CRC resulted in synergistic TGI in the combination arm (61%) relative to monotherapy azenosertib (17%) or adagrasib (2%). Finally, analysis of biomarkers from both in vitro and in vivo tumor samples demonstrated synergistic increases in protein markers of RS, DNA damage, and apoptosis with combination therapy. Taken together, our results suggest that the combination of azenosertib with KRASG12C inhibitors enhances tumor growth inhibition over single agent therapy and may be an effective treatment option for patients with KRASG12C tumors. Citation Format: Nathan M. Jameson, Gabriel Kim, Catherine Lee, Blake Skrable, Alexandra Shea, Monah Abed, Olivier Harismendy, Jianhui Ma, Doris Kim, Mark R. Lackner. The selective WEE1 inhibitor azenosertib shows synergistic anti-tumor activity with KRASG12C inhibitors in multiple KRASG12C models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6487.

Abstract 613: Oral CHK1 inhibitor BBI-355 allows flexibility of dose and schedule with demonstration of monotherapy and combinational antitumor activity in extrachromosomal DNA (ecDNA) driven preclinical models

Abstract Focal, high copy number amplifications of prevalent oncogenes (e.g., MYC, EGFR) frequently occur on extrachromosomal DNA (ecDNA), highly transcribed units of circular, non-chromosomal DNA. While use of targeted therapies is associated with improved survival in patients with cancers that have activating oncogene mutations or fusions, they have demonstrated limited activity for patients whose tumors harbor oncogene amplifications, including those mediated by ecDNA. We have previously identified Checkpoint Kinase 1 (CHK1) as a target essential for ecDNA function in cancer, as tumor cells with oncogene amplification on ecDNA exhibit increased DNA replication stress (RS) and are hypersensitive to inactivation of CHK1, cells’ master regulator of RS. Thus, we have developed BBI-355, a novel, orally bioavailable, and selective small molecule inhibitor of CHK1, which is currently being studied in the first-in-human Phase 1/2 POTENTIATE clinical trial for patients with cancer harboring oncogene amplifications, including on ecDNA (NCT05827614). In vitro antiproliferation activity of BBI-355 was observed in a panel of ecDNA+ oncogene amplified tumor lines and non-amplified lines, with heightened sensitivity and induction of RS observed in ecDNA+ cells. Oral administration of BBI-355 resulted in dose dependent on-target activity, with induced p-CHK1-S345 protein expression, in agreement with measured plasma and tumor concentrations. Subsequent pharmacokinetic/pharmacodynamic (PK/PD) modeling guided continuous and intermittent dose/schedules of single agent BBI-355 that demonstrated durable antitumor activity, including tumor regressions, in multiple ecDNA+ tumor models. Single agent targeted therapy treatment of cancers with oncogene amplification on ecDNA can result in acquired resistance via ecDNA-based mechanisms, further increasing reliance on CHK1. Enhanced antitumor activity of BBI-355 in combination with agents targeting the protein products of amplified oncogenes was demonstrated in multiple in vivo tumor models; these included combination with a pan-FGFR inhibitor in FGFR2 ecDNA+ amplified gastric cancer and combination with CDK4/6 inhibitors in CDK4 ecDNA+ amplified sarcoma models. The first ecDNA-directed therapy (ecDTx) in development, BBI-355 demonstrated significant antitumor activity as a single agent and in combination with targeted therapies in multiple ecDNA+ oncogene amplified tumor models. Oral dosing of BBI-355 provides dose/schedule flexibility to attempt to optimize antitumor activity with a tolerable safety profile in patients with highly aggressive cancers and significant unmet need. The POTENTIATE clinical trial is open and currently enrolling. Citation Format: Ryan J. Hansen, Auzon Steffy, Joshua Plum, Ardalan Ardeshiri, Edison Tse, Salvador Garcia, Ben Norman, Jafar Moininazeri, AnneMarie Pferdekamper, Evan Holmes, Debbie Liao, Rachelle Elsdon, Joshua Lange, Anthony Pinkerton, Julius Apuy, Sudhir Chowdhry, Christian Hassig, Shailaja Kasibhatla. Oral CHK1 inhibitor BBI-355 allows flexibility of dose and schedule with demonstration of monotherapy and combinational antitumor activity in extrachromosomal DNA (ecDNA) driven preclinical models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 613.

Abstract 3998: Preclinical efficacy of a synthetic Notch-based combinatorial immunotherapy against colorectal cancer with HER2 amplification

Abstract HER2 amplification occurs in about 5% of colorectal cancers (CRC) and is only partially associated with clinical response to combined HER2/EGFR targeted treatment. An alternative approach, based on Adoptive Cell Therapy (ACT) using T cells engineered with anti-HER2 Chimeric Antigen Receptor (CAR), proved toxic due to "on-target off-tumor" activity. Therefore, we developed a combinatorial ACT strategy to safely target HER2 amplification in CRC using a synNotch-based artificial regulatory network. A synthetic Notch receptor was employed in which the extracellular domain is an anti-HER2 scFv and the intracellular domain contains the GAL4VP64 artificial transcription factor. Engagement of the HER2-synNotch by overexpressed HER2 on the surface of target cells induces GAL4VP64 cleavage and translocation to the nucleus, where it drives expression of a CAR under a GAL4-responsive promoter. In this way, only cells co-expressing both HER2 and the CAR target are killed. Carcinoembryonic antigen (CEA) was selected as a colon-specific CAR target: CEA expression is indeed restricted to the digestive tract and is increased in CRC. The natural killer cell line NK-92 was chosen as effector to be engineered. NK-92 cells transduced with HER2-synNotch and inducible CEA-CAR lentiviral vectors were subsequently sorted in the OFF and ON state to select those with the higher CAR induction after synNotch engagement. Subsequently, cloning of sorted cells led to identification of an optimally responsive clone (clone 5F), in terms of specificity and amplitude of CAR induction. The 5F clone demonstrated significant activity in vitro and in vivo specifically against HER2amp CRC models, with no effects on cells with physiological HER2 levels. In vitro, the 5F clone displayed selective cytotoxicity against HER2amp/CEA+ CRC cells, with minimal killing activity against HER2amp/CEA- cells, or HER2norm/CEA+ cells. Additional assays on 3D organoids highlighted better recruitment and infiltration by the 5F clone respect to NK-92 WT cells, only in HER2amp models. In vivo, NK-92.5F significantly impaired tumor growth in two different HER2amp CRC models, with no effect on HER2norm CRC cells. To further improve survival, tumor penetration and in vivo efficacy of the NK-92.5F clone, a more complex system was built, in which HER2 synNotch engagement drives expression of both the CEA-CAR and IL2. 5F-IL2 cells displayed further increased cytotoxicity in vitro. In vivo, 5F-IL2 cells drastically increased survival of mice carrying HER2amp CRC xenografts with respect to the parental 5F clone. The observed selective therapeutic efficacy both in vitro and in vivo of this innovative HER2 synNotch/CEA-CAR system, and its future evolutions, opens perspectives for possible clinical applications in cases of HER2amp CRC resistant or partially responsive to HER2/EGFR blockade. Citation Format: Marco Cortese, Erica Torchiaro, Alice D'Andrea, Consalvo Petti, Federica Invrea, Letizia Franco, Chiara Donini, Valeria Leuci, Simonetta M. Leto, Valentina Vurchio, Francesca Cottino, Claudio Isella, Sabrina Arena, Elisa Vigna, Andrea Bertotti, Livio Trusolino, Dario Sangiolo, Enzo Medico. Preclinical efficacy of a synthetic Notch-based combinatorial immunotherapy against colorectal cancer with HER2 amplification [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3998.

Improving the Quality of Warehousing Processes in the Context of the Logistics Sector

Logistics processes play a crucial role in ensuring the smooth operation of the supply chain, and warehouse management is one of the key logistics activities of invaluable significance. To evaluate the management principles of companies, it is necessary to thoroughly analyze their functions, operational standards, and quality requirements. One of the most common strategies in modern businesses is quality improvement. Insufficient quality can lead to competitive disadvantages and have negative consequences for companies that do not prioritize quality and neglect their services and products, resulting in a loss of customer loyalty and revenue. Effective warehouse management is critically important for companies to meet consumer needs, reduce costs, and gain a competitive advantage. Conducting qualitative research is necessary in order to learn how to properly organize warehouse management within companies, which will also help to identify key issues and determine areas for change. This article consists of a theoretical section that analyzes the scientific literature on the quality of warehouse process management in the logistics sector. It examines theoretical aspects of logistics processes and quality assurance in company management, including the implementation of systems that ensure quality in warehouse processes and logistics process management. The subsequent sections describe the research methods applied, evaluate the results of warehouse management quality, and provide insights and recommendations.

Unravelling the role of work function difference on supported Ru nanoclusters for alkaline hydrogen evolution reaction

Fine tuning the work function difference (ΔΦ) between support and Ru which in turn affects the electron distribution of Ru is an effective method for the design of high-performance alkaline hydrogen evolution reaction (HER) catalysts. However, it remains a challenge to understand the intrinsic relationship between HER activity and ΔΦ at the atomic level. Herein, taking N doped graphene supported Ru nanoclusters (Ru/CxNy) as the theoretical model, we demonstrate that the HER activity can be correlated with the variation of ΔΦ. For Ru/C50 with smaller ΔΦ, we find that the Ru atoms located at the top layer of Ru nanoclusters (Ru-t) with lower coordination numbers are the main reaction sites. For Ru/C30N20 with larger ΔΦ, the stronger charge redistributions of Ru is observed, and the electron-deficient Ru atoms closing to the interface (Ru-i) are the main reaction sites with significant improvement HER activity. Our findings demonstrate the role of ΔΦ on Ru/CxNy systems for the alkaline HER, which can provide valuable theoretical guidance for the design of high efficient HER catalysts.

Syntheses, Crystal Structures and Antimicrobial Activity of Zinc(II) Complexes Derived from 5-Bromo-2- (((2-piperazin-1-yl)ethyl)imino)methyl)phenol

Three new zinc(II) complexes, [ZnCl2L]·CH3OH (1), [ZnClL(NCS)]·2CH3OH·0.5H2O (2), and [ZnL(NCS)2]·CH3OH·H2O (3), where L is the zwitterionic form of 5-bromo-2-(((2-piperazin-1-yl)ethyl)imino)methyl)phenol, NCS is thiocyanate anion, were facile prepared by reaction of different molar ratio of L, zinc chloride and ammonium thiocyanate in methanol. The complexes were characterized by IR and UV-Vis spectroscopy. Detailed structures of the three complexes were confirmed by single crystal X-ray determination. The Zn atoms in the complexes are in tetrahedral coordination. The Schiff base ligand coordinates to Zn atom through phenolate oxygen atom and amino and imino nitrogen atoms. The remaining two sites are occupied by two Cl for 1, one Cl and one NCS for 2, and two NCS for 3. The compounds show significant antimicrobial activities.

Exploring sustainable management by using green nano-silver to combat three post-harvest pathogenic fungi in crops

Global crop protection and food security have become critical issues to achieve the ‘Zero Hunger’ goal in recent years, as significant crop damage is primarily caused by biotic factors. Applying nanoparticles in agriculture could enhance crop yield. Nano-silver, or AgNPs, have colossal importance in many fields like biomedical, agriculture, and the environment due to their antimicrobial potential. In this context, nano-silver was fabricated by Citrus medica L. (Cm) fruit juice, detected visually and by UV–Vis spectrophotometric analysis. Further, AgNPs were characterized by advanced techniques. UV–Vis spectroscopic analysis revealed absorbance spectra at around 487 nm. The zeta potential measurement value was noted as -23.7 mV. Spectral analysis by FT-IR proved the capping of the acidic groups. In contrast, the XRD analysis showed the Miller indices like the face-centered cubic (fcc) crystalline structure. NTA revealed a mean size of 35 nm for nano-silver with a 2.4 × 108 particles mL−1 concentration. TEM analysis demonstrated spherical Cm-AgNPs with 20–30 nm sizes. The focus of this research was to evaluate the antifungal activity of biogenic AgNPs against post-harvest pathogenic fungi, including Aspergillus niger, A. flavus, and Alternaria alternata. The Cm-AgNPs showed significant antifungal activity in the order of A. niger > A. flavus > A. alternata. The biogenic Cm-AgNPs can be used for the inhibition of toxigenic fungi.

Efficacy of the induced pluripotent stem cell derived and engineered CD276-targeted CAR-NK cells against human esophageal squamous cell carcinoma.

Chimeric antigen receptor natural killer (CAR-NK) cells have been found to be successful in treating hematologic malignancies and present potential for usage in solid tumors. In this study, we created CD276-targeted CAR-expressing NK cells from pluripotent stem cells (iPSC CD276-targeted CAR-NK cells) and evaluated their cytotoxicity against esophageal squamous cell carcinoma (ESCC) using patient-specific organoid (PSO) models comprising of both CD276-positive and CD276-negative adjacent epithelium PSO models (normal control PSO, NC PSO) as well as primary culture of ESCC cell models. In addition, in vitro and in vivo models such as KYSE-150 were also examined. iPSC NK cells and NK-free media were used as the CAR-free and NK-free controls, respectively. The positive CD276 staining was specifically detected on the ESCC membrane in 51.43% (54/105) of the patients of all stages, and in 51.35% (38/74) of stages III and IV. The iPS CD276-targeted CAR-NK cells, comparing with the iPS NK cells and the NK-free medium, exhibited specific and significant cytotoxic activity against CD276-positive ESCC PSO rather than CD276-negative NC PSO, and exhibited significant cytotoxicity against CD276-expressing cultured ESCC cells, as well as against CD276-expressing KYSE-150 in vitro and in BNDG mouse xenograft. The efficacy of the iPSC CD276-targeted CAR-NK cells demonstrated by their successful treatment of CD276-expressing ESCC in a multitude of pre-clinical models implied that they hold tremendous therapeutic potential for treating patients with CD276-expressing ESCC.

Flavonoids from Liriodendron chinense Leaves Alleviate Oxidative Activity in Vitro and D-galactose-induced Brain Injury via AMPK/SIRT1 Pathway in Vivo

The exploration of active ingredients and new activities of industrial crops could expand their scope of applications. Liriodendron chinense (L. chinense), an important ornamental and economic forest tree in China, has been widely used in timber, medicine and landscape. Flavonoids are major constituents of L. chinense leaves, while their effects of preventing brain damage are still unclear. In this study, flavonoids from L. chinense leaves (LCLF) were obtained through AB-8 resin chromatography and UPLC-MS/MS assay indicated that 16 compounds were identified from LCLF. LCLF treatment showed significant scavenging activities against DPPH•, ABTS•+, •OH, FRAP and total reducing power. D-galactose-impaired exploratory behaviors, cognitive functions and histopathological alterations in hippocampus of mice were obviously recovered by LCLF administration. Network pharmacology results demonstrated that LCLF could produce the healing efficacy through multiple pathways, mostly related to oxidative stress and inflammation. Further in vivo experiments revealed that LCLF treatment could regulated MDA content, SOD, CAT and GSH-Px activity, as well as the production of inflammatory factors (NO, IL-1β and IL-6). The underlying mechanisms of LCLF were probably related to Nrf2/HO-1, NF-κB and AMPK/SIRT1 pathways. Molecular docking results and surface plasmon resonance analysis validated that especially hesperidin in LCLF had strong binding activities with all 8 genes in the mentioned pathways. Our study revealed the beneficial effects of LCLF on oxidative activity and D-galactose-induced brain injury, offering experimental support for its potential application in pharmaceutical and other industries.