Dual Targeting Research Articles

Design, Synthesis and Biological Evaluation of C3‐Indolyl/(3‐chloro‐indolyl)‐C4‐aryl/heteroaryl‐azetidin‐2‐ones

AbstractHerein, trans‐ and cis‐azetidin‐2‐ones 3–6 were strategically synthesized, capitalizing on the bioactivity of azetidin‐2‐ones and indole pharmacophore, followed by a comprehensive characterization using a diverse array of spectroscopic techniques. The sixteen azetidin‐2‐ones were examined for antimicrobial activities against both Gram‐negative (P. aeruginosa, E. coli, A. baumannii) and Gram‐positive bacteria (S. aureus, E. faecium, B. cereus), as well as against C. albicans and C. tropicalis fungal strains. The highly potent compounds (5 a, 6 b, 6 d) demonstrated maximum inhibition against all multidrug‐resistant strains, with minimum inhibitory concentrations ranging from 0.97–3.9 μg/mL, surpassing the potency of standard ampicillin (MIC: 3.12–50 μg/mL). Moreover, 6 b and 6 d exhibited significant inhibitory effects on C. albicans (MIC: 0.97 μg/mL), comparable to fluconazole. The presence of C3‐(3‐chloro‐indolyl) scaffold, combined with diverse electronic effects at N1/C4‐centers, particularly the inclusion of thiophen‐2‐yl motif, greatly influenced the activity of target compounds. Assessment of 4 d, 4 i–k and 6 d on THLE‐2 cell lines revealed their preferential safety. Molecular docking studies revealed seven compounds with active dual targeting of DNA GyrB and PBP2a proteins, demonstrating a potent broad‐spectrum antibacterial effect. In silico ADME analysis affirms positive drug‐likeness and favorable pharmacokinetic characteristics of indole‐derived hybrids, indicating a promising potential for addressing challenges in evolving multidrug resistance.

Dual inhibition of ATR and DNA-PKcs radiosensitizes ATM-mutant prostate cancer.

In advanced castration resistant prostate cancer (CRPC), mutations in the DNA damage response (DDR) gene ataxia telangiectasia mutated ( ATM ) are common. While poly(ADP-ribose) polymerase inhibitors are approved in this context, their clinical efficacy remains limited. Thus, there is a compelling need to identify alternative therapeutic avenues for ATM mutant prostate cancer patients. Here, we generated matched ATM-proficient and ATM-deficient CRPC lines to elucidate the impact of ATM loss on DDR in response to DNA damage via irradiation. Through unbiased phosphoproteomic screening, we unveiled that ATM-deficient CRPC lines maintain dependence on downstream ATM targets through activation of ATR and DNA-PKcs kinases. Dual inhibition of ATR and DNA-PKcs effectively inhibited downstream γH2AX foci formation in response to irradiation and radiosensitized ATM-deficient lines to a greater extent than either ATM-proficient controls or single drug treatment. Further, dual inhibition abrogated residual downstream ATM pathway signaling and impaired replication fork dynamics. To circumvent potential toxicity, we leveraged the RUVBL1/2 ATPase inhibitor Compound B, which leads to the degradation of both ATR and DNA-PKcs kinases. Compound B effectively radiosensitized ATM-deficient CRPC in vitro and in vivo , and impacted replication fork dynamics. Overall, dual targeting of both ATR and DNA-PKcs is necessary to block DDR in ATM-deficient CRPC, and Compound B could be utilized as a novel therapy in combination with irradiation in these patients.

Fluorescent DNA tetrahedral probe with catalytic hairpin self-assembly reaction for imaging of miR-21 and miR-155 in living cells.

ACHA-based fluorescent DNA tetrahedral probe (FDTp) has been designedto detect the microRNAsmiR-21 and miR-155 sensitively and specifically in living cells. The design consisted of functional elements (H1, H2, and Protector) connected to a DNA tetrahedron modified with two pairs of fluorophores and quenching groups. In the presence of miR-21, the chain displacement effect was triggered and Cy3 fluorescence was emitted. In the presence of miR-155, the signal of the catalytic hairpin assembly (CHA) between H1 and H2 on FDTp was amplified, making the fluorescence of FAM sensitive to miR-155. Using this method, the detection limit for miR-155 was 5pM. The FDTp successfully imaged miR-21 and miR-155 in living cells and distinguished a variety of cell lines based on their expression levels of miR-21 and miR-155. The detection and imaging of dual targets in this design ensured the accuracy of tumor diagnosis and provided a new method for early tumor diagnosis.

An ultra-sensitive electrochemical biosensor for circulating tumor DNA utilizing dual enzyme-assisted target recycle and hybridization chain reaction amplification strategies

Circulating tumor DNA (ctDNA) has emerged as a pivotal biomarker for cancers, offering a non-invasive means of detection through liquid biopsies. However, the current methodologies for ctDNA detection often struggle with sensitivity, especially when confronted with ultra-low concentrations. To address this challenge, we have engineered a remarkably sensitive electrochemical biosensor that can detect low concentration of ctDNA. This breakthrough is achieved by integrating a dual enzyme-assisted amplification mechanism with hybridization chain reaction (HCR). In the initial phase, the presence of ctDNA triggers the unfolding of the H2 hairpin structure, resulting in a segment of double-stranded DNA. The subsequent introduction of Klenow (3′→5′ exo-) and nicking endonuclease (Nb.BbvCI) enzymes leads to the abundant production of extended DNA (EXTDNA). This step critically enhances the initial amplification of the target DNA, ensuring superior specificity and selectivity thanks to the synergistic enzymatic activity. EXTDNA efficiently unfolds the H1 hairpin structure immobilized on the gold electrode, which acts as the initiation point for the HCR. Further amplification of the trace target DNA is achieved through an additional H3 and H4 hybrid-mediated HCR. Finally, the differential pulse voltammetry signal of methylene blue is increased significantly. Our results reveal that the developed electrochemical biosensor displays an exceptional linear correlation with ctDNA across concentrations ranging from 10 fM to 20 pM, with an unprecedented detection limit of 2.3 fM (3 s/k). This innovative approach shows immense potential for the ultrasensitive detection of ctDNA, promising broad applications in early cancer detection and monitoring.

Dual targeting non-overlapping epitopes in HER2 domain IV substantially enhanced HER2/HER2 homodimers and HER2/EGFR heterodimers internalization leading to potent antitumor activity in HER2-positive human gastric cancer

BackgroundTrastuzumab and pertuzumab combination has been approved for the treatment of patients with HER2-positive metastatic breast cancer. However, trastuzumab and pertuzumab combination did not show improvement in overall survival in patients with HER2-positive metastatic gastric cancer.MethodsWe developed a new HER2-targeted monoclonal antibody, HLX22, targeting HER2 subdomain IV as trastuzumab but with non-overlapping epitopes. We examined the antitumor effects of this novel HER2-antibody in gastric cell lines and cell line-derived xenograft (CDX) and patient-derived xenograft (PDX) models.ResultsHLX22 in combination with HLX02 (trastuzumab biosimilar) induced enhancement of HER2/HER2 homodimers and HER2/EGFR heterodimers internalization, which ultimately led to the reduction in signal transductions involving STAT3, P70 S6, and AKT; gene expressions of FGF-FGFR-PI3K-MTOR, EGF-EGFR-RAS, TGF-β-SMAD, PLCG and cell cycle progression related pathways that favor tumor development, proliferation, progression, migration and survival in gastric cancer cell line NCI-N87 were also reduced. These differing but complementary actions contributed to the synergistic antitumor efficacy of the HLX22 and HLX02 combination in gastric cancer cell lines, CDX and PDX. In addition, HLX22 in combination with HLX02 demonstrated stronger antitumor efficacy than HLX02 and HLX11 (a potential pertuzumab biosimilar) combination treatment both in vitro and in vivo.ConclusionsThese results suggested that the application of non-competing antibodies HLX22 and HLX02 targeting HER2 subdomain IV together may be of substantial benefit to gastric cancer patients who currently respond suboptimal to trastuzumab therapy.

Delivery of piperlongumine via hyaluronic acid/phenylboronic acid-mediated dual targetable polymersome for enhanced anticancer functionality against pancreatic tumor

Pancreatic cancer cells highly resistance to conventional chemo drugs, resulting low survival rates. The aim of the study was to design and develop dual targeting polymersomes (DTPS) loaded with phyto alkaloid agent i.e., piperlongumine (PL) for effective pancreatic cancer treatment. Here, hyaluronic acid (HA) was functionalized with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPEPEG-NH2), poly(ethylene glycol) bis (amine) (PEG), and phenylboronic acid (PBA) moieties. The designed DTPS could selectively recognize CD44/sialic acid (SA) and deliver PL to MIA PaCa-2 pancreatic cancer cells, facilitated via HA-CD44 and PBA-SA interactions. Drug release and stability results implied sustained PL release profile and pH sensitivity. DTPS could be more efficiently bound with SA than other sugars based on fluorescence spectroscopy. The anticancer efficacy of designed polymersomes was tested with H6C7 normal pancreas cells and SA/CD44-overexpressed MIA PaCa-2 pancreatic cancer cells. DTPS showed both SA and CD44-mediated higher cellular uptake while single-targeted polymersomes showed CD44-mediated cellular uptake. The PL-loaded DTPS efficiently uptake by MIA PaCa-2 cancer cells, causing up to 80 % cell growth inhibition, reduced cell spheroids volume and increased dead cells by 58.3 %. These results indicate that the newly developed DTPS can effectively serve as a pH-responsive drug delivery system for efficient treatment of cancer.

Simultaneous dual-target magnetic resonance-guided focused ultrasound treatment for patients with tremor-dominant Parkinson’s disease

Introduction. Non-invasive magnetic resonance-guided focused ultrasound (MRgFUS) is a new neurosurgical treatment option for tremor-dominant Parkinson’s disease (TDPD). Outcomes of ablation with dual targeting of two subcortical nuclei to improve functional treatment results are yet to be explored. Aim. This study aimed to evaluate the safety and efficacy of MRgFUS with simultaneous unilateral ablation of two cerebral targets in patients with TDPD. Materials and methods. A total of 82 TDPD patients (20 women, 62 men; median age 65.0 [52.5; 70,0] years) received unilateral MRgFUS, i.e. ventrointermedial (VIM) nucleus thalamotomy and/or pallidothalamotractotomy (PTT). Motor symptoms, including tremor, were assessed using MDS-Unified Parkinson’s Disease Rating Scale Part III (MDS-UPDRS-III). VIM, PTT, and VIM + PTT ablation was received by 34, 12, and 36 patients, respectively. Results. After surgery, MDS-UPDRS-III score improved by 40.1% (30.2; 51.7) without early or late-onset serious complications. Tremor returned in 18 patients (all after VIM thalamotomy); 9 of them successfully underwent re-treatment 9–12 months after the first procedure. Simultaneous dual-target (VIM + PPT) intervention was successfully received by 36 patients without any serious complications. A total of 89.3% and 69.7% of patients remained relapse-free in the dual-target and single-target groups, respectively (p = 0.039). Conclusion. Simultaneous dual-target (VIM and PTT) MRgFUS showed favorable safety and efficacy profiles and can be considered a symptomatic treatment option for TDPD patients.

Can carbon emission trading policy enhance the synergistic emission reduction of carbon dioxide and air pollutants? A comparative study considering different pollutants

Exploring the synergistic control of carbon dioxide and pollutants is significant to the dual carbon target. This study first evaluates the effect of CET on the synergistic emission reduction of carbon dioxide and different pollutants. Moreover, this study investigates the mechanisms of synergistic emission reduction through the pathways of technology progress and structure adjustment. Technology progress is categorized into technology research and development, technology introduction, reverse technology spillover and technology purchase. Structure adjustment contains industrial structure and energy structure. Finally, the impact of CET on the synergistic emission reduction in the surrounding regions is analyzed. The results indicate that the CET could promote the synergistic emission reduction of carbon dioxide and pollutants, including air pollutants (SO2, PM2.5), wastewater and solid waste. The synergistic emission reduction of CO2 and SO2 could be enhanced through reverse technology spillover, technology purchase and industrial structure adjustment. The synergistic emission reduction of CO2 and PM2.5 could be enhanced through R&D and reverse technology spillover. The synergistic emission reduction of carbon dioxide and ammonia nitrogen could be enhanced by increasing R&D, technology purchase and energy structure adjustment. The CET could improve the synergistic emission reduction effect of carbon dioxide and air pollutants in the surrounding regions.

Research on Zoning and Carbon Sink Enhancement Strategies for Ecological Spaces in Counties with Different Landform Types

Ecological carbon sinks, pivotal in mitigating carbon emissions, are indispensable for climate change mitigation. Counties, as the fundamental units of ecological space management, directly impact the achievement of regional dual carbon targets through their levels of carbon sink. However, existing research has overlooked the intricate relationship between terrain features and ecological spaces, leading to a lack of specific guidance on enhancing the carbon sink for counties with diverse landform characteristics. This study focused on Jingbian County (Loess Plateau), Fuping County (Guanzhong Plain), and Chenggu County (Qinba Mountains), each characterized by distinct landform characteristics. This study proposes a comprehensive identification model for ecological space within the context of dual carbon targets. Utilizing this model as a basis, the land use structure, carbon sink potential, and ecological spatial patterns of different counties were systematically analyzed. The results indicated substantial disparities in land use structure, carbon sink capabilities, and ecological space distributions among counties with different landform types. Specifically, Jingbian County was predominantly covered by grassland, exhibiting a moderate overall carbon sink capacity, with baseline ecological spaces playing a significant role. Conversely, Fuping County, dominated by cultivated land and construction land, exhibited the lowest carbon sink capacity, with non-ecological spaces accounting for a staggering 85.93%. Chenggu County, on the other hand, was characterized by the dominance of forestland, with nearly all its carbon sink originating from forestland, and core ecological spaces occupying a leading position. Tailored optimization strategies are recommended based on varying terrain features: Jingbian County should prioritize ecosystem restoration and conservation, while Fuping County should concentrate on optimizing land use structure and promoting urban greening. Reinforcing the carbon sink capacity of existing ecosystems is crucial for Chenggu County. This study broadens the perspective on ecological space optimization and provides scientific guidance and pragmatic insights tailored to regional disparities, which are instrumental in assisting various regions to achieve their dual carbon targets.

Energy-Consuming Right Trading Policy and Corporate ESG Performance: Quasi-Natural Experimental Evidence from China

The energy-consuming right trading policy (ECRTP) represents a significant initiative to promote the sustainable development of Chinese enterprises. This study employs the difference-in-differences methodology to analyze how ECRTP influences ESG performance based on data from A-share listed industrial enterprises in China from 2006 to 2020. The findings indicate that ECRTP effectively enhances corporate ESG performance, and this conclusion holds valid following a battery of robustness checks. Moreover, ECRTP improves corporate ESG performance by promoting green technological innovation and green perceptions among executives. Tests for heterogeneity show that the ECRTP exerts a more pronounced influence on ESG performance for enterprises located in regions with high public environmental awareness, heavily polluting industries, and coastal areas. This study broadens the literature on ECRTP effectiveness evaluation, providing valuable insights for refining the design of these policies, promoting their implementation, and facilitating the achievement of dual control targets for energy consumption.

Lineage-dependence of the neuroblastoma surfaceome defines tumor cell state-dependent and independent immunotherapeutic targets.

Neuroblastoma is a heterogeneous disease with adrenergic (ADRN)- and therapy resistant mesenchymal (MES)-like cells driven by distinct transcription factor networks. Here, we investigate the expression of immunotherapeutic targets in each neuroblastoma subtype and propose pan-neuroblastoma and cell state specific targetable cell-surface proteins. We characterized cell lines, patient-derived xenografts, and patient samples as ADRN-dominant or MES- dominant to define subtype-specific and pan-neuroblastoma gene sets. Targets were validated with ChIP- sequencing, immunoblotting, and flow cytometry in neuroblastoma cell lines and isogenic ADRN-to-MES transition cell line models. Finally, we evaluated the activity of MES-specific agents in vivo and in vitro . Most immunotherapeutic targets being developed for neuroblastoma showed significantly higher expression in the ADRN subtype with limited expression in MES-like tumor cells. In contrast, CD276 (B7-H3) and L1CAM maintained expression across both ADRN and MES states. We identified several receptor tyrosine kinases (RTKs) enriched in MES-dominant samples and showed that AXL targeting with ADCT-601 was potently cytotoxic in MES-dominant cell lines and showed specific anti-tumor activity in a MES cell line-derived xenograft. Immunotherapeutic strategies for neuroblastoma must address the potential of epigenetic downregulation of antigen density as a mechanism for immune evasion. We identified several RTKs as candidate MES-specific immunotherapeutic target proteins for the elimination of therapy-resistant cells. We hypothesize that the phenomena of immune escape will be less likely when targeting pan-neuroblastoma cell surface proteins such as B7-H3 and L1CAM, and/or dual targeting strategies that consider both the ADRN- and MES-cell states. Cellular plasticity influences the abundance of immunotherapeutic targets.Subtype-specific targets may be susceptible to epigenetically-mediated downregulation.Immunotherapeutic targets in development, B7-H3 and L1CAM, show "pan-subtype" expression. Neuroblastoma is a lethal childhood malignancy that shows cellular plasticity in response to anti-cancer therapies. Several plasma membrane proteins are being developed as immunotherapeutic targets in this disease. Here we define which cell surface proteins are susceptible to epigenetically regulated downregulation during an adrenergic to mesenchymal cell state switch and propose immunotherapeutic strategies to anticipate and circumvent acquired immunotherapeutic resistance.

A Dual-Targeting, Multi-Faceted Biocompatible Nanodrug Optimizes the Microenvironment to Ameliorate Abdominal Aortic Aneurysm.

Abdominal aortic aneurysm (AAA) is a highly lethal cardiovascular disease that currently lacks effective pharmacological treatment given the complex pathophysiology of the disease. Here, single-cell RNA-sequencing data from patients with AAA and a mouse model are analyzed, which reveals pivotal pathological changes, including the M1-like polarization of macrophages and the loss of contractile function in smooth muscle cells (SMCs). Both cell types express the integrin αvβ3, allowing for their dual targeting with a single rationally designed molecule. To this end, a biocompatible nanodrug, which is termed EVMS@R-HNC, that consists of the multifunctional drug everolimus (EVMS) encapsulated by the hepatitis B virus core protein modifies to contain the RGD sequence to specifically bind to integrin αvβ3 is designed. Both in vitro and in vivo results show that EVMS@R-HNC can target macrophages as well as SMCs. Upon binding of the nanodrug, the EVMS is released intracellularly where it exhibits multiple functions, including inhibiting M1 macrophage polarization, thereby suppressing the self-propagating inflammatory cascade and immune microenvironment imbalance, while preserving the normal contractile function of SMCs. Collectively, these results suggest that EVMS@R-HNC presents a highly promising therapeutic approach for the management of AAA.

Dual-Targeted Zeolitic Imidazolate Frameworks Drug Delivery System Reversing Cisplatin Resistance to Treat Resistant Ovarian Cancer.

Ovarian cancer cells are prone to acquire tolerance to chemotherapeutic agents, which seriously affects clinical outcomes. The development of novel strategies to enhance the targeting of chemotherapeutic agents to overcome drug resistance and minimize side effects is significant for improving the clinical outcomes of ovarian cancer patients. We employed folic acid (FA)-modified ZIF-90 nanomaterials (FA-ZIF-90) to deliver the chemotherapeutic drug, cisplatin (DDP), via dual targeting to improve its targeting to circumvent cisplatin resistance in ovarian cancer cells, especially by targeting mitochondria. FA-ZIF-90/DDP could rapidly release DDP in response to dual stimulation of acidity and ATP in tumor cells. FA-ZIF-90/DDP showed good blood compatibility. It was efficiently taken up by human ovarian cancer cisplatin-resistant cells A2780/DDP and aggregated in the mitochondrial region. FA-ZIF-90/DDP significantly inhibited the mitochondrial activity and metastatic ability of A2780/DDP cells. In addition, it effectively induced apoptosis in A2780/DDP cells and overcame cisplatin resistance. In vivo experiments showed that FA-ZIF-90/DDP increased the accumulation of DDP in tumor tissues and significantly inhibited tumor growth. FA-modified ZIF-90 nanocarriers can improve the tumor targeting and anti-tumor effects of chemotherapeutic drugs, reduce toxic side effects, and are expected to be a novel therapeutic strategy to reverse drug resistance in ovarian cancer.

Mitigating stuttering self-stigma: How do we start and where do we go? Using a Participative Concept Mapping Approach to develop a local framework of principles

PurposeThis study aims to create a stigma reduction framework for stuttering in the local context of Québec, Canada using the Participative Concept Mapping Approach (PCMA), focusing on both self and societal stigma. MethodEmploying a mixed-methods approach, this study engaged 17 experts—people who stutter, clinicians and health innovation specialists—in PCMA workshops. Via diverse steps, including generation, sorting and rating of ideas in response to the focus prompt, "To effectively address stuttering (self-)stigma, an intervention should…" these sessions led to a framework depicted in visual maps, then refined into actionable principles through qualitative analysis. Mixed-methods data analysis used the open-source R-CMap software to generate visual maps illustrating the relationships among ideas as well as importance and feasibility ratings. ResultsThe collaborative workshops identified 95 ideas in response to the focus prompt, reunited in 7 clusters, evolving into 16 principles to mitigate stuttering stigma and self-stigma. At the therapy level, these principles emphasize personalized therapy, thorough assessments, stigma-free therapeutic environment, empowerment, and the importance of group inclusivity and educating the relational circles. Societally, they advocate for initiatives such as improved educational outreach, empathy enhancement, and better representation. This dual approach targets individual experiences and societal views on stuttering, stressing the need for an all-encompassing intervention framework. ConclusionThe findings demonstrate PCMA's usefulness in crafting local, culturally sensitive, tailored interventions for stigma reduction. The study emphasizes the necessity of holistic approaches that address individual experiences and societal perceptions, offering a model to conduct similar exercises in diverse local settings.

Rooftop PV Development Suitability and Carbon Benefits: An Anhui Province Case Study

As one of the most rapidly developing provinces in China in the past two decades, Anhui Province has seen an increasing demand for clean energy in recent years due to industrial transformation and the requirements of dual carbon targets. This paper opts to investigate roof-mounted distributed photovoltaics, which are more suitable for development in densely populated areas. Current research on distributed photovoltaics largely focuses on vague estimations of power generation potential, without adequately considering the specific development conditions of different regions. This paper starts from the actual situation affecting the development of roof-mounted distributed photovoltaics and selects a smaller number of factors that are more in line with reality for hierarchical analysis, constructing a relatively simple but practical evaluation system (“meteorological-geographical-socio-economic”). At the same time, this paper innovatively proposes different schemes for the full lifecycle power generation and emission reduction benefits of roof-mounted distributed photovoltaics and compares them, providing a foundation for subsequent in-depth research. Key findings include the following: The northern regions of Anhui Province exhibit higher suitability for rooftop distributed PV, with residential areas being the primary influencing factor, followed by solar radiation considerations; the annual power generation potential of rooftop distributed PV in Anhui Province constitutes around 80% of the total electricity consumption in 2021, but the potential is predominantly concentrated in rural areas, resulting in spatial disparities in power generation and consumption across the province; developing the rooftop distributed PV industry based on suitability can yield substantial power generation and emission reduction benefits, translating to an estimated reduction of approximately 1.28 × 108 tCO2 annually, representing around one-third of Anhui Province’s carbon emissions in 2021.

Dual-template magnetic molecularly imprinted polymers for selective extraction and sensitive detection of aflatoxin B1 and benzo(α)pyrene in environmental water and edible oil

The coexistence of multiple contaminates in the environment and food is of growing concern due to their extremely hazard as a well-known class I carcinogen, like aflatoxin B1 (AFB1) and benzo(α)pyrene (BaP). AFB1 and BaP are susceptible to coexistence in environmental water and edible oil, posing a significant potential risk to environmental monitoring and food safety. The remaining challenges in detecting multiple contaminates include unsatisfied sensitivity, insufficient targets selectivity, and interferences in complex matrices. Here, we developed dual-template magnetic molecularly imprinted polymers (DMMIPs) for selective extraction of dual targets in complex matrices from the environment and food. The DMMIPs were fabricated by surface imprinting with vinyl-functionalized Fe3O4 as carrier, 5,7-dimethoxycoumarin and pyrene as dummy templates, and methacrylamide as functional monomer. The DMMIPs showed excellent adsorption ability (12.73–15.80 mg/g), imprinting factors (2.01–2.58), and reusability of three adsorption-desorption cycles for AFB1 and BaP. The adsorption mechanism including hydrogen bond, electrostatic interaction and van der Waals force was confirmed by physical characterization and DFT calculation. Applying DMMIPs in magnetic solid phase extraction (MSPE) followed by high-performance liquid chromatography (HPLC) analysis enabled detection limits of 0.134 μg/L for AFB1 and 0.107 μg/L for BaP. Recovery rates for water and edible oil samples were recorded as 86.2%–110.3% with RSDs of 4.1%–11.9%. This approach demonstrates potential for simultaneous identification and extraction of multiple contaminants in environmental and food.

RETRACTED: Hemiprotonic ph-ph+ with two targets inhibits metastatic breast cancer and concurrent candidiasis

Concurrent infection in breast cancer patients is the direct cause of the high mortality rate of the disease. However, there is no available method to increase the survival rate until now. To address the problem, we propose one drug with two target strategy to treat the refractory disease. A small chemical, ph-ph+, was attempted to be used in the study to explore the feasibility of the approach in anticancer and antifungus at the same time. The results showed that ph-ph+ could prevent the proliferation and metastasis of breast cancer cells, and kill C. albicans simultaneously. The molecular mechanism was associated with the activation of an evolutionarily conserved protease CLpP in the cancer and C. albicans cells. Also, the signaling pathway mediated by PLAGL2 that highly expressed in cancer cells participated in preventing cell metastasis and inducing apoptosis of ph-ph+. The one drug with dual targets inhibited the growth and metastasis of the cancer cells, and meanwhile eliminated C. albicans in tissues in the experimental animals. The results suggested that ph-ph+ with dual targets of CLpP and PLAGL2 would be a feasible approach to prolong the survival rate in patients with metastatic breast cancer and pathogenic infection.

ASAH1 facilitates TNBC by DUSP5 suppression-driven activation of MAP kinase pathway and represents a therapeutic vulnerability

Triple-negative breast cancer (TNBC) is a subtype of breast cancer that is prone to metastasis and therapy resistance. Owing to its aggressive nature and limited availability of targeted therapies, TNBC is associated with higher mortality as compared to other forms of breast cancer. In order to develop new therapeutic options for TNBC, we characterized the factors involved in TNBC growth and progression. Here, we demonstrate that N-acylsphingosine amidohydrolase 1 (ASAH1) is overexpressed in TNBC cells and is regulated via p53 and PI3K-AKT signaling pathways. Genetic knockdown or pharmacological inhibition of ASAH1 suppresses TNBC growth and progression. Mechanistically, ASAH1 inhibition stimulates dual-specificity phosphatase 5 (DUSP5) expression, suppressing the mitogen-activated protein kinase (MAPK) pathway. Furthermore, pharmacological cotargeting of the ASAH1 and MAPK pathways inhibits TNBC growth. Collectively, we unmasked a novel role of ASAH1 in driving TNBC and identified dual targeting of the ASAH1 and MAPK pathways as a potential new therapeutic approach for TNBC treatment.

Discovery of Novel Small Molecule Dual Inhibitor Targeting Toll-Like Receptors 7 and 9.

The aberrant secretion of proinflammatory cytokines by immune cells is the principal cause of inflammatory diseases, such as systemic lupus erythematosus and rheumatoid arthritis. Toll-like receptor 7 (TLR7) and TLR9, sequestered to the endosomal compartment of dendritic cells and macrophages, are closely associated with the initiation and progression of these diseases. Therefore, the development of drugs targeting dysregulated endosomal TLRs is imperative to mitigate systemic inflammation. Here, we applied the principles of computer-aided drug discovery to identify a novel low-molecular-weight compound, TLR inhibitory compound 10 (TIC10), and its potent derivative (TIC10g), which demonstrated dual inhibition of TLR7 and TLR9 signaling pathways. Compared to TIC10, TIC10g exhibited a more pronounced inhibition of the TLR7- and TLR9-mediated secretion of the proinflammatory cytokine tumor necrosis factor-α in a mouse macrophage cell line and mouse bone marrow dendritic cells in a concentration-dependent manner. While TIC10g slightly prevented TLR3 and TLR8 activation, it had no impact on cell surface TLRs (TLR1/2, TLR2/6, TLR4, or TLR5), indicating its selectivity for TLR7 and TLR9. Additionally, mechanistic studies suggested that TIC10g interfered with TLR9 activation by CpG DNA and suppressed downstream pathways by directly binding to TLR9. Western blot analysis revealed that TIC10g downregulated the phosphorylation of the p65 subunit of nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) and mitogen-activated protein kinases (MAPKs), including extracellular-signal-regulated kinase, p38-MAPK, and c-Jun N-terminal kinase. These findings indicate that the novel ligand, TIC10g, is a specific dual inhibitor of endosomal TLRs (TLR7 and TLR9), disrupting MAPK- and NF-κB-mediated proinflammatory gene expression.

Rapid and Amplification-free Nucleic Acid Detection with DNA Substrate-Mediated Autocatalysis of CRISPR/Cas12a.

To enable rapid and accurate point-of-care DNA detection, we have developed a single-step, amplification-free nucleic acid detection platform, a DNA substrate-mediated autocatalysis of CRISPR/Cas12a (DSAC). DSAC makes use of the trans-cleavage activity of Cas12a and target template-activated DNA substrate for dual signal amplifications. DSAC employs two distinct DNA substrate types: one that enhances signal amplification and the other that negatively modulates fluorescent signals. The positive inducer utilizes nicked- or loop-based DNA substrates to activate CRISPR/Cas12a, initiating trans-cleavage activity in a positive feedback loop, ultimately amplifying the fluorescent signals. The negative modulator, which involves competitor-based DNA substrates, competes with the probes for trans-cleaving, resulting in a signal decline in the presence of target DNA. These DNA substrate-based DSAC systems were adapted to fluorescence-based and paper-based lateral flow strip detection platforms. Our DSAC system accurately detected African swine fever virus (ASFV) in swine's blood samples at femtomolar sensitivity within 20 min. In contrast to the existing amplification-free CRISPR/Dx platforms, DSAC offers a cost-effective and straightforward detection method, requiring only the addition of a rationally designed DNA oligonucleotide. Notably, a common ASFV sequence-encoded DNA substrate can be directly applied to detect human nucleic acids through a dual crRNA targeting system. Consequently, our single-step DSAC system presents an alternative point-of-care diagnostic tool for the sensitive, accurate, and timely diagnosis of viral infections with potential applicability to human disease detection.