Modern high-density neural recordings demand spike sorting algorithms that can handle diverse probe geometries and complex, neuron-specific drift, yet existing methods often rely on rigid geometric assumptions and one-dimensional drift models. Here, we introduce KIASORT (Knowledge-Integrated Automated Spike Sorting), a geometry-free approach for per-neuron drift tracking. KIASORT builds channel-specific sorting models from a hybrid linear-nonlinear sample-sorting stage, using representative template banks or supervised classifiers. These channel-specific models then sort spikes by independently tracking each neuron, unconstrained by probe layout. Biophysical simulations showed that even sub-micron probe displacements induce neuron-specific waveform distortions that standard drift models cannot correct. In ground-truth benchmarks with heterogeneous, neuron-specific drift, KIASORT outperformed Kilosort4 in recovering high-quality units, while maintaining real-time performance on standard CPUs. Its robustness was further illustrated on both primate and mouse data. KIASORT combines automated sorting with manual curation in a unified graphical interface, offering a complete and user-friendly spike sorting platform. The software is freely available at https://kiasort.com.Significance Statement Accurate spike sorting remains a fundamental challenge in systems neuroscience, particularly as recording technologies advance toward simultaneous monitoring of thousands of neurons and the next generations of recording probes. Current methods often rely on rigid assumptions about probe geometry and uniform drift patterns for different neurons which often fail in real-world recordings. We introduce KIASORT with capability to track neurons in a geometry-free framework, as a fundamentally new approach that addresses these critical limitations with many use cases which are not supported by other existing methods.
Single-walled carbon nanotubes (SWCNT) are versatile building blocks for optical sensors. Their near infrared photoluminescent emission is determined by their chiral structure. Commercially available SWCNT are polydisperse, leading to spectral congestion and no ability single analyte sensing. Aqueous two-phase extraction (ATPE) is a technique to sort SWCNT structures for improved nanosensor performance and multiplexed sensing. To date, ATPE has not been demonstrated with bioconjugate-compatible surface chemistries. Here, we demonstrate improved and multiplexed cytokine sensing by ATPE sorting SWCNT species with amine-functionalized ssDNA. We show that this approach enables effective chirality sorting and direct antibody conjugations. We used sorted, functionalized SWCNT to simultaneously and specifically detect the inflammatory cytokines IL-12 and IL-6 with high sensitivity and selectivity, and improved robustness compared to unsorted SWCNT. We anticipate that such sensitive, multiplexed will be useful for implantable and bedside diagnostics of inflammatory processes while serving as a model for sensing other complex biological processes.
Spatial mode sorting has become a prominent optical processing modality due to its capability of saturating fundamental limits to several sensing tasks, including wavefront sensing, exoplanet detection, and superresolution imaging. Yet despite these potential sensitivity advantages, contemporary mode sorters often feature large crosstalk, high loss, or sort modes that are poorly adapted to conventional imaging systems (e.g., Hermite- and Laguerre-Gauss). Because most imaging and sensing systems are circularly symmetric, an arguably more prudent strategy is to instead demultiplex modes that are natural to circular optics, namely the Zernike polynomials. To realize this goal, we propose a mode sorting architecture that relies on a cascade of vortex-phase Fourier filters and Mach-Zehnder interferometers. When these filters' vortex charges are appropriately chosen, we show that it is possible to assemble a series of vortex-phase Fourier filters that can isolate the various Zernike modes losslessly and without crosstalk. Our idea is demonstrated via applications to phase estimation and exoplanet imaging, where we propose an optical system that saturates the quantum sensitivity limits to both tasks. We therefore anticipate that this work will be of practical value for the high-contrast imaging of extrasolar planets, enhancing both wavefront control and coronagraph performance.
Nucleoli, nuclear speckles and other compartments regulate transcription, RNA processing, and chromatin organization within the nucleus, yet the relationship of their morphology to developmental gene expression programs in vivo is poorly understood. Here, we develop a high-throughput Visual Cell Sorting (VCS) workflow for fixed cells and nuclei that combines antibody-based photoconversion; GPU-accelerated, real-time image analysis; and three-level single-cell combinatorial indexing RNA-seq (sci-RNA-seq3) to link nuclear compartment morphology to single-nucleus transcriptomes at embryo scale. We use VCS to analyze and sort over 1 million mouse embryo-derived nuclei by nucleolar, nuclear speckle, or nuclear size and construct a transcriptional atlas annotated with nuclear compartment phenotypes. Nuclear compartment size varies both between and within lineages and is shaped by proliferation and differentiation. In extracellular matrix protein-producing cell types such as fibroblasts, chondrocytes, and osteoblasts, nucleolar enlargement is uncoupled from cell cycle, and in erythroid cells exhibit a sharp nucleolar contraction preceding cell-cycle exit. We identify a 41-gene transcriptional signature whose expression tracks nucleolar size, enriched for ribosome biogenesis, mitochondrial metabolism, unfolded protein response, stress granule, and ubiquitin-proteasome pathway components. We used this nucleolar transcriptional signature to annotate mouse, zebrafish and human developmental atlases with nucleolar size information, revealing a conserved coupling between nucleolar activity and proteostasis programs. Our work establishes Visual Cell Sorting as a scalable platform for mapping image-based phenotypes to molecular programs; details the relationship between nuclear compartment phenotypes and development; and provides a transcriptional signature to estimate nucleolar size from existing single-cell datasets.
Fluorescence-activated cell sorting (FACS) has been a core technology for the identification, characterization and isolation of hematopoietic stem cells (HSCs), allowing for prospective isolation of cells which express specific molecules at specific levels of intensity. A recent technological breakthrough, image-enabled cell sorting (ICS), is based on FACS but it also permits separation of cells based on the differential distribution of these molecules across the cell. Using ICS in primary HSCs, we surveyed the patterns of expression for a range of intracellular markers and identified mitochondrial dyes as a tool to prospectively isolate HSCs with different properties. Integration of index-sort data with single-cell divisional kinetics identified total intensity and diffusivity of the MitoTracker dye as parameters enabling prospective isolation of functionally heterogeneous HSCs. In vivo functional assays confirmed that HSCs capable of long-term multilineage contribution were highly enriched in the MitoTracker low diffusivity, low intensity populations. Moreover, HSCs displaying high MitoTracker intensity but low MitoTracker diffusivity (Mito±), displayed the characteristics of low MitoTracker intensity (Mito-/-), representing a subpopulation of HSCs only isolatable by tools such as ICS. Collectively these data demonstrate that ICS is a powerful tool for prospectively enriching highly specific cell populations in the hematopoietic system, making it broadly useful for applications where population identification and purity are required to attain robust cellular and molecular characterization.
Autistic adults self-harm more often than non-autistic adults, but there are no tools to assess patterns of self-harm in autistic people. This study aimed to (1) review the accessibility of a novel visual task - the Card Sort Task for Self-Harm (CaTS) - to explore self-harm with autistic adults and (2) to pilot the CaTS to identify proximal and distal contributing factors leading to self-harm and experiences following self-harm. First, autistic adults (n = 5) with lived experience of self-harm reviewed the CaTS for clarity and accessibility. Second, we undertook a pilot administration of the CaTS and used sequence analysis to explore significant transitions between factors before and after self-harm. Participants (n = 29, autistic or possibly autistic, female = 82%, mean age = 41.62) selected, on average, 42 cards to describe self-harm. Most frequently selected cards described agitation (n = 25), mental pain (n = 22) and depression (n = 22), while most infrequently collected cards included being in a gang and talking to a teacher (n = 0). Agitation and acting on impulse preceded self-harm. Feeling better, worse, exhausted and hopeless followed self-harm. Improving emotion regulation and meeting autistic communication needs could reduce the transition to self-harm. The CaTS offers a systematic approach to explore self-harm in autistic adults.Lay AbstractWhy did we do this study?Autistic adults self-harm more often than people who are not autistic. This is particularly worrying because self-harm is something that can contribute to suicide. We know that autistic people feel that their experience of self-harm is not well understood by other people, such as doctors.What did we want to know?We wanted to know if the card sort task for self-harm (called the CaTS for short) is a useful way to explore self-harm with autistic adults.What did we do?First, we worked with five autistic adults to make sure the CaTS is clear and works for autistic people. Second, we invited autistic and non-autistic adults to do the CaTS. To do the CATS, someone chooses the cards that are relevant to their experience and puts them on a timeline to explain what self-harm is like for them.What did we find?We found that (1) the CaTS is helpful and accessible to explore self-harm with autistic adults and (2) the CaTS uncovered patterns of self-harm. Twenty-nine U.K.-based autistic adults did the CaTS: most were female (82%, average age was around 42). Participants picked, on average, 42 cards to describe self-harm. The cards that were chosen most often described agitation, mental pain and depression. The cards that were chosen least described being in a gang and talking to a teacher. The order of the cards suggested that people felt agitated and acted impulsively before self-harm. After self-harm, they felt better, worse and hopeless. We found that it is safe and feasible to do the CaTS with autistic people. Support could consider how best to support people who are impulsive and how to prevent people accessing the means to hurt themselves.
Heat stress occurs when total environmental and metabolic heat production is greater than an animal's ability to dissipate that heat. Heat stress negatively impacts feeder cattle performance and welfare. Limited research has been conducted to determine if feeding cattle in the evening, thereby shifting their metabolic heat production to a cooler period of the day, can help mitigate heat stress. This pen-level randomized controlled trial evaluated the effects of evening feeding (PM; feedings at 2000, 2300, and 0200 h; n = 24 pens) versus morning feeding (AM; feedings at 0500, 0800, and 1200 h; n = 24 pens) on terminally sorted steer performance in a commercial feedyard in the Pacific Northwest. Data collection included feed delivery, water consumption, health events, open mouth breathing prevalence, and carcass traits. Linear and generalized linear mixed-effects models were used to determine potential differences between treatment group and temperature-humidity index (THI; <80 versus ≥80). Only 14% of the total study days had a THI ≥ 80, indicating little to no heat stress impacts. There were no differences found between PM and AM for any outcome (p < 0.05). Regardless of treatment group, water consumption tended (p = 0.07) to increase, and open mouth breathing significantly (p < 0.05) increased on days with THI ≥ 80. Further research is warranted to assess evening feeding as a heat stress mitigation strategy in a feedyard setting.
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Breast reconstruction improves body image, sexual experience, and well-being in patients with breast cancer. However, only about 10% of patients in China choose breast reconstruction. The objective of this study is to explore types of breast reconstruction decision-making and driving factors among Chinese premenopausal patients. This was a mixed-methods study based on Q-methodology, conducted from May 2024 to October 2024. First, 15 premenopausal breast cancer patients who underwent breast reconstruction were selected using maximum variation purposive sampling for semistructured interviews, generating a Q-sample dataset consisting of 36 statements. Then, 18 premenopausal patients who underwent breast reconstruction were selected as the P-sample using the same sampling method; these 18 patients performed Q-sorting on the 36 statements within a specified Q-sorting grid and completed in-depth interviews after sorting. Data analysis was primarily conducted using Ken-Q software, which automatically generated standardized Q-sort results. Combined with these standardized Q-sort results and interview contents, the study analyzed the decision-making driving types and factors influencing patients' choices of breast reconstruction. Principal component analysis identified six factors with eigenvalues > 1, collectively accounting for 72% of the total variance. Based on the ideal Q-sort simulated by the software, three types of decision-making factors were identified. The social participation and youth advantage co-dominant type was motivated by professional advice, younger age, and social engagement. The aesthetic appearance and case-based encouragement co-dominant type was driven by a desire for body image, aesthetics, and the influence of other's successful surgeries. The medical professional advice and financial status co-dominant type was shaped by trust in medical professionals, confidence in the healthcare team, and financial considerations. Premenopausal patients with breast cancer in China who underwent breast reconstruction demonstrated various decision-making types. The findings may assist Chinese healthcare providers as well as healthcare providers in other countries with similar Eastern cultural backgrounds in understanding and exploring the drivers of breast reconstruction choices during shared decision-making processes. This work can help in developing targeted interventions and establishing clinical protocols that empower patients to make choices that align with their preferences and best interests.
Despite many available treatments for female chronic pelvic pain (FCPP), there is limited patient success, engagement, or compliance with therapy. No current methods allow providers to effectively collect and integrate patient perspectives into treatment choice for female chronic pelvic pain (FCPP). We aimed to leverage former qualitative stakeholder input on FCPP treatment choice, patient advisory group (PAG) input, and Q-methodology to translate patient viewpoints around FCPP treatment into measurable patient viewpoints to guide treatment choice. We used Q-methodology to translate qualitative data formerly collected from stakeholders in FCPP treatment into measurable patient preference profiles. PAG members interpreted previously collected qualitative data from FCPP stakeholders, and their guidance formulated Q-set statements in English and Spanish. Patients with FCPP performed Q-sort of these statements. PAG input then guided interpretation of factor analyses of Q-sorts to form relevant patient preference profiles for FCPP and provided preliminary mapping to certain treatments that may resonate with patients aligned with each preference profile. We engaged 11 PAG members, and 47 patients completed Q-sort. The authors and the PAG analyzed, interpreted, and described five factors (patient preference profiles) to measure FCPP treatment viewpoints. We titled these patient preference profiles as the "Self-empowered Realist," the "Fearless Escalator," the "Careful Scientist," the "Trusting Optimist," and "Eager to Move On," with each patient having a certain alignment with each of these viewpoints. These factors map to possible treatments that align with a patient's viewpoint as they align with one or more of these patient preference profiles. We described the perspectives associated with each patient preference profile to allow for future use of these profiles to inform instruments to measure patient preferences in FCPP treatment navigation. We translated patient perspectives on FCPP treatment into five measurable, nuanced patient preference profiles. We plan to use these patient preference profiles to guide treatment option counseling for FCPP in future studies.
Cloth-liquid interaction involves coupled phenomena such as porous flow, absorption, emission, and diffusion, which are difficult to model in a stable and efficient manner in particle-based simulations. In this paper, we present a GPU-based cloth-liquid coupling framework that integrates these processes within a unified Smoothed Particle Hydrodynamics (SPH) pipeline. To improve directional robustness in porous flow near boundaries, we use a virtual-pressure formulation inspired by Darcy's law, while absorption and emission are handled through saturation-based mass exchange and diffusion is applied as a sequential redistribution stage. For efficient neighbor search, the framework employs a Bitonic sort-based GPU hashing structure. We evaluate the proposed method using qualitative wet-cloth scenarios and quantitative analyses including frame-time measurement, sorting-cost comparison, scalability with increasing particle counts, mass conservation error, and module-wise timing breakdown. The results show that the proposed hashing scheme reduces sorting overhead compared with the baseline GPU implementation, while the full coupling pipeline maintains stable wetting behavior and bounded mass variation across the tested scenes. These results indicate that the proposed framework provides an efficient and numerically stable approach for GPU-based porous cloth-liquid simulation.
Neoadjuvant chemotherapy (NAC) followed by definitive treatment consisting of either radical radiotherapy or radical cystectomy is the recommended treatment for patients with organ-confined muscle-invasive bladder cancer (OC-MIBC). A randomised controlled trial (RCT) aimed to compare the effectiveness of radical radiotherapy and radical cystectomy but failed to recruit. Radical radiotherapy is non-invasive and organ-preserving, and observational studies have suggested this treatment may be associated with similar outcomes compared with radical cystectomy. However, in these observational studies, the risk of confounding was high, and they did not consider the receipt of NAC. The surgery or radiotherapy (SORT) for the early-stage cancer study will assess the comparative effectiveness and cost-effectiveness of either radical cystectomy or radical radiotherapy, both after NAC for OC-MIBC. We will use a target trial emulation approach to reduce the risk of bias when assessing comparative effectiveness from observational data. The SORT study will use UK's National Cancer Registry to identify individuals diagnosed with urothelial OC-MIBC (T2-4aN0M0) between 1 January 2015 and 31 December 2021 who received either radical radiotherapy or radical cystectomy after NAC. The data will be linked to Hospital Episode Statistics (HES), National Radiotherapy Data Set (RTDS) and Systemic Anti-Cancer Therapy (SACT) data sets to gather information on clinical, tumour and socio-demographic characteristics and receipt of treatment. Using the target trial emulation framework, we will define the eligibility criteria and radical radiotherapy and radical cystectomy receipt. To reduce the risk of confounding, we will use advanced statistical approaches to allow for differences in measured baseline characteristics between the comparison groups.The primary outcome is 3-year all-cause mortality after radical treatment receipt. Secondary outcomes will include all-cause and bladder-cancer-associated mortality at 3 and 5 years, time to death, incremental costs and incremental cost-effectiveness reported according to net health benefits. The study was approved by the London School of Hygiene and Tropical Medicine Ethics Committee (Reference number 29717 - 1). Results will be communicated in open-access journals and conferences to clinicians, researchers, patients and policymakers.
Congenital syphilis (CS) has been rapidly re-emerging as a public health crisis across North America. The present standard of care treatment for CS involves a 10-day hospitalisation for intravenous penicillin G, which is costly for healthcare systems and families. Given these costs and the frequency of their application, the purpose of this study was to locate the evidence upon which the standard of care for CS has been founded and to assess its quality.A search of Embase and MEDLINE was conducted, and a total of 398 publications were evaluated for inclusion in this study. 54 publications met the inclusion criteria. Quality of data was appraised using the Oxford Centre for Evidence-Based Medicine (OCEBM) levels of evidence 2011 and the American Academy of Family Physicians grades of strength of recommendation taxonomy (SORT). Levels of evidence in OCEBM 2011 are assigned a number from 1 (highest quality) to 5 (weakest). The strength of recommendations in SORT is assigned a letter from A (strongest recommendation) to C (weakest). Level 5 evidence and grade C recommendations were the most abundant results in this study. Additionally, the most common treatment regimens reported were intramuscular penicillin administration, in doses of 50 000-75 000 units/kg, for a duration of 10-14 days. Systematic statistical analyses could not be applied to the results due to inconsistency and arbitrary changes of treatment within the individual constituent studies. The results affirmed a gap in high-quality primary evidence behind the treatment of CS. Supporting literature is based on expert opinion, consensus and non-randomised studies. Overall, this project underscores the need for high-quality randomised controlled trials to understand the limitations of the current standard of care and to confirm the need for a 10-day course of intravenous penicillin G in hospital.
Background: Culturally and Racially Marginalised (CaRM) communities in Australia encounter subtle and covert forms of prejudice, commonly referred to as "new racism". Within healthcare settings, these experiences can shape trust, engagement, and patterns of help-seeking. Mental health nurses are often the first point of contact in care delivery, and their ability to recognise, respond to, and mitigate the impacts of new racism is critical for fostering therapeutic relationships and supporting equitable access. Understanding how CaRM communities perceive the conditions that influence their mental health service use is fundamental for informing more equitable and culturally responsive care. Objective: This study explored the viewpoints of CaRM community members regarding the factors they consider important for addressing new racism in healthcare systems and supporting engagement with mental health services. Design: Q methodology was used to identify statistically derived viewpoints that reflect shared viewpoints about the conditions perceived as critical for addressing the impacts of new racism on mental health service use. Setting: Participants were recruited from culturally and linguistically diverse communities across Australia through community settings, social media, and professional networks. Participants: Thirty-five individuals from CaRM backgrounds completed the Q-sort. Methods: This Q methodology consisted of five steps: (1) set up of the Q-sorting instrument, (2) selection of participants, (3) data collection, (4) factor analysis, and (5) factor interpretation. Results: Three distinct viewpoints were identified: (1) raising awareness of mental health issues within CaRM communities (community-focused), (2) providing visible anti-racism and culturally safe services (service-focused), and (3) recognising and formally addressing new racism within healthcare systems (policy-focused). Conclusions: This study offers the first empirically derived, community-informed set of viewpoints on addressing new racism in Australian mental healthcare. While exploratory, the findings highlight multi-level considerations that are potentially relevant to mental health nursing practice, and may be useful to inform future research, policy development, and service redesign aimed at strengthening cultural responsiveness and equity in mental health systems.
Cognitive flexibility is defined as the capacity to adjust thoughts and behaviors in accordance with goals and changing situational demands. Across all included studies, it was operationalized using neuropsychological tasks, such as the Wisconsin Card Sorting Test, Dimensional Change Card Sort, and Flanker/More-Odd Shifting Task. This study aims to investigate the distinct effects of different exercise prescription parameters (i.e., duration, frequency, period, intensity, and type) on cognitive flexibility in children and adolescents, with the ultimate goal of providing evidence to inform the development of targeted interventions for enhancing cognitive flexibility in this population. This systematic review and network meta-analysis was prospectively registered in PROSPERO (CRD42024539167). Following the PICOS principle, we established inclusion and exclusion criteria and searched the EBSCO, PubMed, Web of Science, Embase, and Cochrane databases through November 15, 2025. Eligible studies enrolled children and adolescents aged 4-18 years, compared any exercise intervention with usual physical activity, and reported cognitive flexibility outcomes. Two reviewers independently screened titles and abstracts, extracted data, and assessed the risk of bias. Network meta-analysis was performed using Stata 17.0, and SUCRA values were used to rank the effect sizes of different exercise prescription parameters. A total of 21 articles involving 3099 participants were included in the study. The network meta-analysis revealed that interventions lasting 31-50 min were significantly more effective than those lasting 10-30 min [SMD=-0.79, 95% CI (-1.55, -0.03)] and those lasting ≥ 71 min [SMD=-1.30, 95% CI (-2.35, -0.25)]. Single aerobic exercise [SMD = 0.67, 95% CI (0.09, 1.25)] and cognitive+exercise interventions [SMD = 0.64, 95% CI (0.09, 1.18)] were significantly more effective than the control group. Additionally, interventions lasting 5-8 weeks were significantly more effective than those lasting ≥ 20 weeks [SMD=-0.79, 95% CI (-1.55, -0.03)]. Among the various intervention characteristics examined, cumulative ranking probability analysis (SUCRA) suggested that sessions lasting 31-50 min (SUCRA = 98.3%), administered twice weekly (SUCRA = 98.4%), implemented over 5-8 weeks (SUCRA = 91.4%), at low intensity (SUCRA = 89.3%), and involving ball games (SUCRA = 98.4%) may hold the greatest probability of being the most effective approach for improving cognitive flexibility in children and adolescents. SUCRA rankings suggest that exercise sessions of 31-50 min, delivered twice per week for 5-8 weeks at low intensity and incorporating ball games, are most likely to yield the greatest benefits for cognitive flexibility. However, this reflects ranking probability rather than effect size magnitude. Direct pairwise comparisons indicated only modest effect sizes, highlighting the need for additional randomized controlled trials to confirm both the relative superiority of these parameters and their clinical significance.
Cells employ lysosomal degradation to eliminate extracellular and membrane proteins, which is a process that can be exploited for therapeutic purposes. Current lysosome-targeting chimeras (LYTACs) that hijack endogenous lysosome-associated receptors (LARs) to degrade pathogenic proteins are constrained by their reliance on endogenous receptor trafficking. Here we introduce synthetic chimeric antigen lysosome-associated receptors (CALARs) that reprogram cells to effectively sort extracellular proteins to lysosomes, operating independently of native ligand competition. Encoded within circular RNA, CALARs enable sustained suppression of immune checkpoints and growth factor receptors for over 1 week. This prolonged suppression enhances immune activation and substantially reduces the tumor burden in vivo. This study introduces a modular, genetically encodable platform for modulating protein expression and advancing cell-based therapeutics.
Lipid nanoparticles (LNPs) have revolutionized nucleic acid therapeutics, yet their inherent ApoE-mediated liver tropism remains a formidable barrier to extrahepatic applications. This review critically synthesizes emerging breakthroughs in engineering next-generation LNPs for precision targeting. We first categorize advancements in passive targeting via physicochemical modulation, followed by a rigorous evaluation of active targeting through ligand functionalization (e.g., antibodies, aptamers). Central to this discussion is the ongoing paradigm shift from empirical screening to mechanism-guided design, highlighted by the "Selective Organ Targeting" (SORT) framework, endogenous protein corona programming, and novel ionizable lipid architectures. These strategies have enabled potent delivery to diverse extrahepatic niches, including the lungs, spleen, and tumors. Furthermore, we address persistent translational hurdles, such as the "PEG dilemma" in ligand conjugation and the clinical unpredictability of the protein corona. By integrating AI-driven discovery with multi-strategy synergy, the field is moving toward a future of programmable, cell-specific therapeutics. This review provides a roadmap for navigating the complexities of non-viral vector design in the post-vaccine era.
Complexes of the μ-oxo-bis(μ-acetate)diruthenium class exhibit ligand lability at the positions trans to the μ-oxo bridge. Lability is an important property for numerous drugs, such as cis-[Pt(NH3)2Cl2], given that its biological activity is related to an aquation reaction taking place in the biological environment. Inspired by that, we have synthesized, characterized, and explored the lability of the new complex [Ru2(μ-O)(μ-CH3COO)2(py)4(thiq)2](PF6)2 (1, py = pyridine; thiq = 5,6,7,8-tetrahydroisoquinoline). The aquation rates in water, phosphate, and tris-HCl buffered solutions were in the 10-4 s-1 range. Complex 1 quenched HSA fluorescence predominantly through a dynamic mechanism (kq = 1.74 to 2.50 × 1012 M-1 s-1, at 298 to 322 K, pH 7.2 tris-HCl buffer), and the HSA excited state lifetime changed markedly. Circular dichroism data showed that the HSA secondary structure did not change in the presence of complex 1. We rationalized these observations in terms of its hydrophilicity (logP = -0.42 ± 0.04), which probably prevented it from accessing the HSA inner hydrophobic pockets. On the basis of our data, lability in complex 1 did not favor any sort of interaction. Compared to the chemically inert [Ru3(μ-O)(μ-CH3COO)6(thiq)3]PF6 compound, complex 1 presented lower cytotoxicity against B16F10 and MCF7 cells. We assigned the small cytotoxic effect to complex 1 releasing the thiq organic ligand during its aquation. Despite not being a good metallodrug, complex 1 is a chemically useful scaffold for exploring ligand-release strategies in biological medium while yielding a non-cytotoxic aqua-complex.
In chronic lymphocytic leukemia (CLL), B-cell receptor, chemokine receptor and integrin signaling contribute to disease progression by influencing cell survival, migration and microenvironment interactions. Focal adhesion kinase (FAK) has recently been involved in these disease processes. Our recent research revealed a correlation between activated FAK and molecules involved in CLL aggressiveness, particularly in IGHV-unmutated cases, including the Lyn kinase substrates HS1 and cortactin. In this context, ROR1 is a key player, sharing connections with Lyn substrates and FAK pathways. We assessed FAK and ROR1 bidirectional association by stimulating CLL cells with Wnt5a and measuring FAK phosphorylation by Western blot, while ROR1 expression was evaluated following FAK inhibitor treatment. Flow cytometry using CXCR4 and CD5 markers was performed to identify and sort proliferating (CXCR4dim/CD5bright) versus quiescent (CXCR4bright/CD5dim) B-cell subpopulations from CLL patients. FAK and ROR1 levels were compared between these fractions. Findings were further supported ex vivo in ibrutinib-treated patients. The FAK inhibitor defactinib was tested alone and in combination with BTK inhibitors on primary CLL cells cultured with or without stromal support, measuring apoptosis by Annexin V/PI staining. We demonstrated that ROR1 triggering by Wnt5a increases FAK activation, while FAK inhibition reduces ROR1 expression. Significantly higher levels of FAK and ROR1 were detected in the proliferating subpopulation corresponding to cells egressing lymph nodes compared to quiescent cells. Ex vivo experiments confirmed high FAK and ROR1 levels in circulating lymphocytes redistributing from secondary lymphoid organs. Defactinib significantly enhanced apoptosis in CLL cells when combined with BTK inhibitors, even in supportive microenvironments, showing significant efficacy also against aggressive CLLs. Our findings reveal a reciprocal regulatory relationship between FAK and ROR1, with both proteins enriched in CLL cells that have exited the lymph nodes. Given defactinib's favorable safety profile in solid tumor trials, combining FAK inhibition with BTK inhibitors could enhance therapeutic efficacy by limiting leukemic clone adaptability and addressing resistance and relapse. While anti-ROR1 therapies showed limited efficacy as monotherapy in CLL trials, targeting multiple nodes of this regulatory network may prove more effective.
As a strategic energy source with huge potential, the commercial development of natural gas hydrate (NGH) is a complex issue concerning the future energy landscape and maritime security. This article aims to systematically sort out the research progress of the entire chain from resource exploration to development and trial production in this field, and dialectically analyze the dual attributes of coexisting energy potential and engineering risks. This article systematically reviews the exploration technology system centered on geophysics, geochemistry, and drilling, as well as the evolution of resource evaluation methods. Furthermore, the support of basic research, such as the hydrate accumulation mechanism and simulation experiments for engineering practice, was explored. By analyzing global trial production cases, the advantages and disadvantages of different mining methods were compared, and core safety challenges, such as geological instability and methane leakage, were distilled. This article ultimately points out that the current development of NGH still faces technical challenges, and the future development will achieve a development system of "multi-technology-driven + multi-objective coordination + intelligent self-adaptation." This article aims to provide a systematic reference framework for the scientific research and commercialization strategy of NGH.