Early Cycle Research Articles

From tissue lesions to neurotoxicity: The devastating effects of small-sized nanoplastics on red drum Sciaenops ocellatus

Nanoplastic pollution typically exhibits more biotoxicity to marine organisms than microplastic pollution. Limited research exists on the toxic effects of small-sized nanoplastics on marine fish, especially regarding their post-exposure resilience. In this study, red drum (Sciaenops ocellatus) were exposed to small-sized polystyrene nanoplastics (30 nm, PS-NPs) for 7 days for the exposure experiments, followed by 14 days of recovery experiments. Histologically, hepatic lipid droplets and branchial epithelial liftings were the primary lesions induced by PS-NPs during both exposure and recovery periods. The inhibition of total superoxide dismutase activity and the accumulation of malondialdehyde content throughout the exposure and recovery periods. Transcriptional and metabolic regulation revealed that PS-NPs induced lipid metabolism disorders and DNA damage during the initial 1–2 days of exposure periods, followed by immune responses and neurotoxicity in the later stages (4–7 days). During the early recovery stages (2–7 days), lipid metabolism and cell cycle were activated, while in the later recovery stage (14 days), the emphasis shifted to lipid metabolism and energy metabolism. Persistent histological lesions, changes in antioxidant capacity, and fluctuations in gene and metabolite expression were observed even after 14 days of recovery periods, highlighting the severe biotoxicity of small-sized PS-NPs to marine fish. In summary, small-sized PS-NPs have severe biotoxicity, causing tissue lesions, oxidative damage, lipid metabolism disorders, DNA damage, immune responses, and neurotoxicity in red drum. This study offers valuable insights into the toxic effects and resilience of small-sized nanoplastics on marine fish.

LiF-Rich Solid Electrolyte Interphase Formation by Establishing Sacrificial Layer on the Separator.

The formation of a stable solid electrolyte interphase (SEI) layer is crucial for enhancing the safety and lifespan of Li metal batteries. Fundamentally, a homogeneous Li+ behavior by controlling the chemical reaction at the anode/electrolyte interface is the key to establishing a stable SEI layer. However, due to the highly reactive nature of Li metal anodes (LMAs), controlling the movement of Li+ at the anode/electrolyte interface remains challenging. Here, an advanced approach is proposed for coating a sacrificial layer called fluorinated self-assembled monolayer (FSL) on a boehmite-coated polyethylene (BPE) separator to form a stable SEI layer. By leveraging the strong affinity between the fluorine functional group and Li+, the rapid formation of a LiF-rich SEI layer in the cell production and early cycling stage is facilitated. This initial stable SEI formation promotes the subsequent homogeneous Li+ flux, thereby improving the LMA stability and yielding an enhanced battery lifespan. Further, the mechanism behind the stable SEI layer generation by controlling the Li+ dynamics through the FSL-treated BPE separator is comprehensively verified. Overall, this research offers significant contributions to the energy storage field by addressing challenges associated with LMAs, thus highlighting the importance of interfacial control in achieving a stable SEI layer.

A novel STPA approach to software safety and security in autonomous maritime systems

With higher autonomy in maritime systems, tasks and responsibilities are moved from the human operator to software, increasing the complexity and the importance of safe and reliable functionality. Software failures, however, may be introduced from the early life cycle phases intentionally or unintentionally, and these must therefore be mitigated by safe and secure design approaches. A challenge is that existing methods are not particularly well-suited for analyzing software risks. Thus, the objective of this paper is to propose a systematic and efficient software failure identification approach by extending the Systems-Theoretic Process Analysis (STPA) with a software failure taxonomy and the System Modeling Language (SysML). This enables the control structure in STPA to cover both the dynamic and static aspects of the software functions. Combined with an implementation platform independent questionnaire, this gives a more systematic and guided search for potential software failures than existing approaches. To demonstrate the proposed approach, a case study on a ferry's navigation system that operates in manual control or semi-autonomous mode is performed. In the case study, the focus is on creating an avoidance map data structure, including both moving and static obstacles to be avoided by the ferry, and the subsequent process of collision risk warning calculation. Software failures are identified and evaluated in collision scenarios where the ferry operates under foggy conditions. The paper shows that the proposed systematic approach provides an improved process for identifying and analyzing critical software failures. This facilitates enhanced risk mitigation in the design and testing phases contributing to autonomous systems' safety and security.

The Impact of Repeated Steam Sterilization Cycles on the Efficacy of Chairside Adjustment Kits for Polishing Monolithic Multi-Layered Zirconia Dental Restoration Material.

BACKGROUND Before insertion, chairside adjustment kits are heat sterilized for positioning and polishing dental restorations. This study aimed to evaluate the effects of 2 steam sterilization cycles on the efficacy of polishing highly translucent monolithic zirconia (HTMLZ) dental restoration material. MATERIAL AND METHODS 100 HTMLZ disc-shaped specimens were adjusted (grinding, finishing, polishing) with EVE Diacera kit. Two steam sterilization techniques [standard (Gp S), immediate/flash (Gp (F)] of CAK were further subgrouped based on number of sterilization cycles [cycle 1 (control), cycle 5, 10, 15, and 20 (experimental)] (n=10 each). Each subgroup accordingly was evaluated for average surface roughness (Ra) and root mean square roughness (Rq) using a profilometer. Mean and standard deviation of 5 subgroups were statistically analyzed using one-way ANOVA/post hoc Tukey's test. Scanning electron microscopy complemented Ra, Rq measurements. Statistical differences of P≤0.05 were considered significant. RESULTS HTMLZ specimens in both groups showed increased (Ra/Rq) values after repeated sterilization of EVE Diacera kit, with Gp F showing lesser increase than Gp S (20 cycles). Gp F at 10 cycles and Gp S at 15 cycles showed clinically unacceptable roughness threshold (0.25 μm). Differences between subgroups for Ra and Rq values were significant (P≤0.05) with less differences within groups observed in early cycles (1, 10). Results validate the manufacturer's recommendations of using flash sterilization/10 cycles for EVE Diacera kit. CONCLUSIONS Repeated sterilization reduces efficacy of chairside adjustment kit to produce smooth surfaces on HTMLZ. This study recommends flash sterilization to a maximum of 10 times to get the clinically acceptable results of Ra and Rq.

Selection based on the phenomenic approach and agronomic ideotic of white oat

AbstractThis work aimed to propose multivariate selection strategies reconciling the agronomic ideotype and vegetation indices to obtain genotypes with high tillering, early cycle, tolerant to lodging, and foliar diseases. The sowing of white oat lines was carried out on April 27, 2023, allocated in an experimental design of blocks augmented with interspersed controls. The regular treatments correspond to 593 lines of white oat and the common treatments were represented by the cultivars URS Olada, URS Altaneira, and IPR Artêmis arranged in four replications. The measurements of variables of agronomic interest took place in each experimental unit as follows: percentage of tillering, days to flowering, percentage of lodging, percentage of leaf rust, percentage of stem rust, and percentage of leaf spots. The phenomic analyses took place through flights carried out with a specialized drone, equipped with a 20 MP resolution camera and standard height (80 m). The best proposed indices were index for reducing the percentage of lodging with normalized green (NG), green–red ratio (IGR), soil color index (SCI), green leaf index (GLI), visible atmospheric resistant index (VARI), green leaf index 2 (GLI2), red–blue (RB), blue green pigment (BGI), normalized red–blue difference index (NRBDI), green–blue ratio, BIM, normalized green–blue difference index (NGBDI), and ExGR; index for reducing days to flowering with brightness index (BI), NRBDI, days to flowering (DF), gray (GRAY), IGR, average intensity (L), normalized red (NR), RGRI, SCI, BGI, BIM, GLI, NGBDI, and NG; and index for reducing the percentage of stem rust with NG, BI, Gray2, GLI2, RGRI, GLI, VARI, BGI, L, NR, RB, and coloration index. Only two lines were selected concomitantly with multivariate indices, line L167 (M5, SA) and L426 (M6, ETMS), that combine lodging tolerance and leaf rust genes.

Survey of Ventilator Waveform Interpretation Among ICU Professionals.

The interpretation of ventilator waveforms is essential for effective and safe mechanical ventilation but requires specialized training and expertise. This study aimed to investigate the ability of ICU professionals to interpret ventilator waveforms, identify areas requiring further education and training, and explore the factors influencing their interpretation skills. We conducted an international online anonymous survey of ICU professionals (physicians, nurses, and respiratory therapists [RTs]), with ≥ 1 y of experience working in the ICU. The survey consisted of demographic information and 15 multiple-choice questions related to ventilator waveforms. Results were compared between professions using descriptive statistics, and logistic regression (expressed as odds ratios [ORs; 95% CI]) was performed to identify factors associated with high performance, which was defined by a threshold of 60% correct answers. A total of 1,832 professionals from 31 countries or regions completed the survey; 53% of respondents answered ≥ 60% of the questions correctly. The 3 questions with the most correct responses were related to waveforms that demonstrated condensation (90%), pressure overshoot (79%), and bronchospasm (75%). Conversely, the 3 questions with the fewest correct responses were waveforms that demonstrated early cycle leading to double trigger (43%), severe under assistance (flow starvation) (37%), and early/reverse trigger (31%). Factors significantly associated with ≥ 60% correct answers included years of ICU working experience (≥ 10 y, OR 1.6 [1.2-2.0], P < .001), profession (RT, OR 2.8 [2.1-3.7], P < .001), highest degree earned (graduate, OR 1.7 [1.3-2.2], P < .001), workplace (teaching hospital, OR 1.4 [1.1-1.7], P = .008), and prior ventilator waveforms training (OR 1.7 [1.3-2.2], P < .001). Slightly over half respondents correctly identified ≥ 60% of waveforms demonstrating patient-ventilator discordance. High performance was associated with ≥ 10 years of ICU working experience, RT profession, graduate degree, working in a teaching hospital, and prior ventilator waveforms training. Some discordances were poorly recognized across all groups of surveyed professionals.

Potential Antitumor Activity of Combined Lycopene and Sorafenib against Solid Ehrlich Carcinoma via Targeting Autophagy and Apoptosis and Suppressing Proliferation.

An FDA-approved kinase inhibitor called sorafenib (SOR) is used to treat primary kidney and liver cancer as well as to stop the spread of advanced breast cancer. Side effects from SOR, such as palmar-plantar erythrodysesthesia syndrome, can negatively impact an individual's quality of life. There are a lot of data supporting the importance of lycopene (LYC) in preventing cancer. The antitumor properties of the combination of sorafenib and lycopene were examined in this study. A viability test against MDA-MB-231 was used to assess the anticancer efficacy of sorafenib, lycopene, and their combination in vitro. Moreover, a cell cycle analysis and Annexin-V/PI double staining were performed by using flow cytometry. In addition, the protein level of JNK-1, ERK-1, Beclin-1, P38, and P53 of the MDA-MB-231 cell line was estimated using ELISA kits. In addition, mice with SEC were divided into four equal groups at random (n = 10) to investigate the possible processes underlying the in vivo antitumor effect. Group IV (SEC-SOR-LYC) received SOR (30 mg/kg/day, p.o.) and LYC (20 mg/kg/day, p.o.); Group I received the SEC control; Group II received SEC-SOR (30 mg/kg/day, p.o.); and Group III received SEC-LYC (20 mg/kg/day, p.o.). The findings demonstrated that the combination of sorafenib and lycopene was superior to sorafenib and lycopene alone in causing early cell cycle arrest, suppressing the viability of cancer cells, and increasing cell apoptosis and autophagy. Likewise, the combination of sorafenib and lycopene demonstrated inhibition of the levels of Bcl-2, Ki-67, VEGF, IL-1β, and TNF-α protein. Otherwise, the quantities of the proteins BAX, P53, and caspase 3 were amplified. Furthermore, the combined treatment led to a substantial increase in TNF-α, caspase 3, and VEGF gene expression compared to the equivalent dosages of monotherapy. The combination of sorafenib and lycopene enhanced apoptosis and reduced inflammation, as seen by the tumor's decreased weight and volume, hence demonstrating its potential anticancer effect.

Longitudinal profiling of human androgenotes through single-cell analysis unveils paternal gene expression dynamics in early embryo development.

How do transcriptomics vary in haploid human androgenote embryos at single cell level in the first four cell cycles of embryo development? Gene expression peaks at the fourth cell cycle, however some androcytes exhibit unique transcriptional behaviors. The developmental potential of an embryo is determined by the competence of the oocyte and the sperm. However, studies of the contribution of the paternal genome using pure haploid androgenotes are very scarce. This study was performed analyzing the single-cell transcriptomic sequencing of 38 androcytes obtained from 10 androgenote bioconstructs previously produced in vitro (de Castro et al., 2023). These results were analyzed through different bioinformatics software such as g: Profiler, GSEA, Cytoscape, and Reactome. Single cell sequencing was used to obtain the transcriptomic profiles of the different androcytes. The results obtained were compared between the different cycles studied using the DESeq2 program and functional enrichment pathways using g: Profiler, Cytoscape, and Reactome. A wave of paternally driven transcriptomic activation was found during the third-cell cycle, with 1128 upregulated and 225 downregulated genes and the fourth-cell cycle, with 1373 upregulated and 286 downregulated genes, compared to first-cell cycle androcytes. Differentially expressed routes related to cell differentiation, DNA-binding transcription, RNA biosynthesis and RNA polymerase II transcription regulatory complex, and cell death were found in the third and fourth with respect to the first-cell cycle. Conversely, in the fourth cell cycle, 153 downregulated and 332 upregulated genes were found compared with third cell cycle, associated with differentially expressed processes related to E-box binding and zinc finger protein 652 (ZNF652) transcription factor. Further, significant overexpression of LEUTX, PRAMEF1, DUXA, RFPL4A, TRIM43, and ZNF675 found in androgenotes, compared to biparental embryos, highlights the paternal contributions to zygote genome activation. All raw sequencing data are available through the Gene Expression Omnibus (GEO) under accessions number: GSE216501. Extrapolation of biological events from uniparental constructs to biparental embryos should be done with caution. Maternal and paternal genomes do not act independently of each other in a natural condition. The absence of one genome may affect gene transcription of the other. In this sense, the haploid condition of the bioconstructs could mask the transcriptomic patterns of the single cells. The results obtained demonstrated the level of involvement of the human paternal haploid genome in the early stages of embryo development as well as its evolution at the transcriptomic level, laying the groundwork for the use of these bioconstructs as reliable models to dispel doubts about the genetic role played by the paternal genome in the early cycles of embryo development. This study was funded by Instituto de Salud Carlos III (ISCIII) through the project 'PI22/00924', co-funded by European Regional Development Fund (ERDF); 'A way to make Europe'. F.D. was supported by the Spanish Ministry of Economy and Competitiveness through the Miguel Servet program (CPII018/00002). M.J.E. was supported by Instituto de Salud Carlos III (PI19/00577 [M.J.E.]) and FI20/00086. P.dC. was supported by a predoctoral grant for training in research into health (PFIS PI19/00577) from the Instituto de Salud Carlos III. All authors declare having no conflict of interest with regard to this trial.

Can primary voters still “kick the tires?” Trends in candidate events in the New Hampshire primary, 2004–2020

ABSTRACT Ample evidence suggests that presidential primaries are fertile ground for personal appearances by candidates to matter, as candidates strategically decide when and where they appear, and voters learn about and may be more likely to vote for candidates they meet. Are recent presidential candidates engaging in less retail politicking in the New Hampshire primary than candidates in earlier cycles? An analysis of 6,079 candidate events from 2004 to 2020 finds that candidates are holding fewer events, that political parties and affiliated groups are sponsoring more events, and that primary events overall are trending away from ones where voters and candidates can interact. While less known candidates still hope an early state focus will catapult them to their party’s nomination, more famous candidates tend to hold fewer events with less opportunity for citizens to interact with them.

An adversarial learning approach to generate pressure support ventilation waveforms for asynchrony detection

Background and objectiveMechanical ventilation is a life-saving treatment for critically-ill patients. During treatment, patient-ventilator asynchrony (PVA) can occur, which can lead to pulmonary damage, complications, and higher mortality. While traditional detection methods for PVAs rely on visual inspection by clinicians, in recent years, machine learning models are being developed to detect PVAs automatically. However, training these models requires large labeled datasets, which are difficult to obtain, as labeling is a labour-intensive and time-consuming task, requiring clinical expertise. Simulating the lung-ventilator interactions has been proposed to obtain large labeled datasets to train machine learning classifiers. However, the obtained data lacks the influence of different hardware, of servo-controlled algorithms, and different sources of noise. Here, we propose VentGAN, an adversarial learning approach to improve simulated data by learning the ventilator fingerprints from unlabeled clinical data. MethodsIn VentGAN, the loss functions are designed to add characteristics of clinical waveforms to the generated results, while preserving the labels of the simulated waveforms. To validate VentGAN, we compare the performance for detection and classification of PVAs when training a previously developed machine learning algorithm with the original simulated data and with the data generated by VentGAN. Testing is performed on independent clinical data labeled by experts. The McNemar test is applied to evaluate statistical differences in the obtained classification accuracy. ResultsVentGAN significantly improves the classification accuracy for late cycling, early cycling and normal breaths (p< 0.01); no significant difference in accuracy was observed for delayed inspirations (p = 0.2), while the accuracy decreased for ineffective efforts (p< 0.01). ConclusionsGeneration of realistic synthetic data with labels by the proposed framework is feasible and represents a promising avenue for improving training of machine learning models.

Developmental progression of DNA double-strand break repair deciphered by a single-allele resolution mutation classifier

DNA double-strand breaks (DSBs) are repaired by a hierarchically regulated network of pathways. Factors influencing the choice of particular repair pathways, however remain poorly characterized. Here we develop an Integrated Classification Pipeline (ICP) to decompose and categorize CRISPR/Cas9 generated mutations on genomic target sites in complex multicellular insects. The ICP outputs graphic rank ordered classifications of mutant alleles to visualize discriminating DSB repair fingerprints generated from different target sites and alternative inheritance patterns of CRISPR components. We uncover highly reproducible lineage-specific mutation fingerprints in individual organisms and a developmental progression wherein Microhomology-Mediated End-Joining (MMEJ) or Insertion events predominate during early rapid mitotic cell cycles, switching to distinct subsets of Non-Homologous End-Joining (NHEJ) alleles, and then to Homology-Directed Repair (HDR)-based gene conversion. These repair signatures enable marker-free tracking of specific mutations in dynamic populations, including NHEJ and HDR events within the same samples, for in-depth analysis of diverse gene editing events.

Abstract 1302: Development and implementation of a systematic approach to measure patient-reported outcomes in early-phase cancer trials

Abstract Background: There has been a shift in the clinical trial landscape toward early-phase studies where data from well-designed and pivotal phase I/II trials is leading to accelerated drug approval. Our goal was to develop a standardized approach to the measurement and reporting of patient-reported outcomes (PROs) in early-phase trials to improve assessment of tolerability, address noted barriers to inclusion, and enhance patient-provider communication to inform treatment decisions and ultimately improve outcomes. Methods: The Office of Patient Centered Outcomes Research (OPCORe) was established in 2022 within the Center for Cancer Research at the National Cancer Institute (NCI) with the mission to advance understanding of the clinical benefit and tolerability of cancer therapies by integrating patient-centered approaches into CCR clinical trials by fostering education and collaboration with stakeholders. Implementation of regulatory/consortia guidelines related to use of core constructs, assessment frequency, and hypothesis-driven objectives and analysis plans through protocol template, development of data capture, email-based alerts and data visualization for clinical providers was developed and standardized templates for protocol language, research staff education, tools for workflow, and tracking implemented. Results: The OPCORe protocol template includes measurement of 3 constructs, including tolerability, clinical benefit and disease-specific impact, as well as anchors of meaningful change. Core symptomatic toxicities are measured to inform understanding across trials with agent/class-specific symptomatic toxicities added to assure relevance. Baseline and standardized weekly electronic capture of tolerability during early cycles with alerts to clinical research staff if moderate-severe toxicities reported helps facilitate patient check-ins. Repeat PROs assessments in later cycles are expanded to include disease-specific assessments and align with disease evaluation to further inform clinical impact. To date, we have collaborated with 7 research groups and developed approaches for first-in-human, phase I, and phase II studies with trial start-up and data collection in process. Conclusion: The value of including PROs in cancer trials is increasingly recognized by regulators, clinicians, patients, and advocates to advance understanding of the clinical benefit &amp; tolerability of emerging therapies. Standardization and incorporation of PROs in earlier phase trials may enable PROs to be optimized, improve later phase trial design, and help inform health policy, support drug approval, and promote shared decision making between providers and patients to inform treatment decisions. Citation Format: Amanda L. King, Jason E. Levine, Elizabeth Vera, Yuti Gandhi, Elaine Novakovich, Theresa Yu, Sefanit Berhanu, Ciara Locke, Kaelyn Julmeus, Tito Mendoza, Terri S. Armstrong. Development and implementation of a systematic approach to measure patient-reported outcomes in early-phase cancer trials [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 1302.

Abstract 5480: Single cell and spatial transcriptomic profile of appendiceal tumour peritoneal disease

Abstract Introduction: Appendiceal tumors (AT) are rare and often develop peritoneal disease (PD), with 5 year survival of 15-63% for high grade adenocarcinomas (HG) and 46-96% for low grade mucinous neoplasms (LG). While the mainstay of treatment is cytoreductive surgery and heated intraperitoneal chemotherapy (CRS/HIPEC), the role of systemic chemotherapy is less clear with limited biological characterization of AT and its tumor microenvironment (TME). The purpose of this study was to evaluate the TME of AT PD. Here we present an atlas of AT PD cells alongside a spatially resolved transcriptomic profile comparing HG and LG. Methods: We collected tissue from 29 patients with AT PD during CRS prior to HIPEC (18 LG, nine HG and two no tumor control). We performed comprehensive single-cell RNA sequencing (38224 cells) on fresh tissue specimens from nine patients (four HG, five LG), and whole transcriptome spatial analysis (&amp;gt;18000 genes) using the GeoMx Digital Spatial Profiler on formalin-fixed paraffin-embedded tissue from 22 patients (five HG, 13 LG). In the spatial profiling, we used anti-CK20, anti-CD45 and SYTO 83 as morphology markers to select areas of interest (AOIs): tumor (CK20), immune (CD45) and stroma (non-CK20/CD45). Analyses were performed with R and a false discovery rate threshold of 0.05. Results: Single-cell RNA sequencing showed that both LG and HG have a preponderance of T-lineage cells including CD8, CD4 and NK T-cells. Both LG and HG tumors display association of mesothelial cells with cancer-associated fibroblasts (CAF), and CAF subsets expressing genes of epithelial-mesenchymal transition (EMT). LG and HG tumors display different patterns of mucin gene expression - epithelial cells of HG express the secreted mucin MUC20 in addition to MUC2 and MUC5B also expressed in LG; a subset of mesothelial cells in LG but not HG express the transmembrane mucin genes MUC1, MUC3A, MUC12 and MUC16. Spatial analysis of tumor AOIs revealed HG compared to LG had increased expression of genes associated with MYC targets, oxidative phosphorylation, anti-tumor immune pathways (IFNα/γ response, intestinal immune network for IGA production), cell cycle pathways (E2F targets, G2M checkpoint) and EMT. LG compared to HG had increased expression of genes associated with inflammation-mediated disease (TNFα signaling via NFKB, inflammatory response), regulation of tumor growth (P53), hypoxia and early cell cycle (estrogen response) and glycosylation pathways. Immune and stromal AOIs did not have significant transcriptomic differences between LG and HG. Conclusion: This study provides novel insights into the biological profile of AT PD by describing TME cell types, immune function, tumor growth and cell cycle pathways. These findings may be used to identify clinically relevant biomarkers and new therapeutic targets. Citation Format: Madeleine C. Strach, Nicole Yeung, Eva Apostolov, Tony Wang, Hui-Ming Lin, Nabila Ansari, Cherry Koh, Joo-Shik Shin, James Kench, Alexander Swarbrick, Lisa Horvath, Kate L. Mahon. Single cell and spatial transcriptomic profile of appendiceal tumour peritoneal disease [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 5480.

Abstract 4681: Unlocking the anticancer potential of calix[4]arene-based Cu(I) ionophores in vitro and in vivo

Abstract Introduction: Cuproptosis is a novel programmed cell death pathway triggered by increased intracellular Copper (Cu) levels. This process offers promising avenues for cancer therapy and underscores the pivotal role of Cu-targeting molecules, such as chelators and ionophores. Despite the progress made, there is still a critical need for novel molecules capable of disrupting copper homeostasis within cancer cells. Methods: We developed and patented a novel class of Cu(I) ionophores characterized by calix[4]arene structures. The anticancer effects of these compounds were evaluated through proliferation assays (IC50), cell cycle assays, and the mechanisms of cell death were explored, including apoptosis, senescence, autophagy, and cuproptosis. To elucidate the mechanisms of action, we conducted proteomic and phosphoproteomic analyses in three different lung adenocarcinoma cell lines (A549, H322, and PC9). Intracellular concentrations of Zn, Fe, and Cu were determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Animal experiments were conducted on nude mice bearing subcutaneous A549 lung cancer xenografts. Results: Our lead compound, Cuphoralix, demonstrated potent cytostatic effect across a panel of 60 cancer cell lines. This effect was attributed to an early and irreversible blockade of the cell cycle initiation at 24 hours post-treatment. Cuphoralix treatment did not induce senescence or apoptosis. Proteomic analyses confirmed the differential regulation—both upregulation and downregulation—of over 500 proteins and phosphopeptides following treatment with Cuphoralix. Notably, Cuphoralix treatment resulted in decreased expression levels of proteins implicated in cell cycle progression and DNA replication. We discovered that Cuphoralix induced cuproptosis and autophagy, as evidenced by the overexpression of LC3B-II. Intracellular Cu level was significantly increased after treatment with Cuphoralix, suggesting the initiation of cuproptosis. In vivo, the toxicity of liposomal Cuphoralix was assessed following three intravenous injections/week for 4 weeks. Animal weights remained stable, with no observed toxicity or signs of neurological or functional impairment. The anticancer efficacy of Cuphoralix, both alone and in combination with other treatments, was corroborated in vivo by measuring the volumes of subcutaneous xenografts. Conclusions: Our novel Cu(I) ionophore, Cuphoralix, has demonstrated significant potential in inducing cuproptosis and disrupting copper homeostasis, presenting a novel approach to cancer therapy. The compound's capacity to initiate an early and irreversible cell cycle blockade and to modulate the expression of key proteins underscores its therapeutic promise. The profound in vitro and in vivo anticancer effects observed encourage further development and investigation of cuproptosis as a targeted cancer treatment strategy. Citation Format: Pierre Lelievre, Cyril Nogier, Nathan Renier, Jean-Luc Coll, Ivan Jabin, Lucie Sancey, Hennie Valkenier, Aurelien Deniaud, Benoit Busser. Unlocking the anticancer potential of calix[4]arene-based Cu(I) ionophores in vitro and in vivo [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 4681.

Sedimentological evolution of the Quibas site: High-resolution glacial/interglacial dynamics in a terrestrial pre-Jaramillo to post-Jaramillo sequence from southern Iberian Peninsula

The sedimentary infill of the Quibas karstic site (Early Pleistocene, southern Spain) represents the only continuous succession with remains of continental vertebrates in Europe from pre-Jaramillo to post-Jaramillo age. The Quibas site, with a significant paleontological record, is dated between 1.1 and 0.9 Ma and offers a unique opportunity to carry out a paleoclimatic reconstruction of the time period immediately after the arrival of the first humans to western Europe. For this reason, defining the dominant sedimentary processes in the different stratigraphic units and the associated paleoenvironment is essential. The Quibas site is made up of two karstic features with two stratigraphic sequences: Quibas-Cueva, containing six lithostratigraphic units, and Quibas-Sima, which contains seven lithostratigraphic units. The detailed description and analyses of the stratigraphic sections have allowed the characterization of various autochthonous and allochthonous facies of cave deposits. Paleoclimatic proxies, inferred from sedimentological analyses, reveal a record of several alternating humid and arid phases resulting from the Early Pleistocene glacial-interglacial cycles, allowing correlation to the marine oxygen isotope record. The lowermost units (pre-Jaramillo) were deposited during a long-lasting interglacial, correlated to MIS 33–31. It was followed by an increase in aridity in the intermediate units of Quibas-Sima and uppermost unit of Quibas-Cueva (Jaramillo), revealing the beginning of a glacial period at the start of the Jaramillo subchron (1 Ma), which can be correlated to MIS 30. The upper Jaramillo and post-Jaramillo units suggest these were deposited in alternating periods of aridity and humid conditions, although less humid than the pre-Jaramillo period, probably representing the MIS 29 interglacial, the MIS 28 glacial and the MIS 27 interglacial.

Over‐ and Hyper‐Lithiated Oxides as Sacrificial Cathodes for Lithium‐Ion Batteries

AbstractBy incorporating sacrificial lithium (Li) sources during electrode fabrication, researchers aim to address the challenge of initial capacity loss due to the formation of a solid electrolyte interphase layer during the early cycles of lithium‐ion batteries (LIBs). This research contributes to the augmentation of Li+ inventory within the electrode to compensate for the irreversible loss of Li+, thereby enhancing the reversibility and cycling performance of LIBs. There are various types of pre‐lithiation additives; however, this perspective specifically discusses over‐ and hyper‐lithiated oxide materials. Within these oxides, research directions are characterized by contrasting approaches aimed at either enhancing the reversibility or inducing the irreversibility of these materials. Intriguingly, both opposing approaches align with the common objective of increasing the energy density of LIBs by providing surplus Li+ to compensate for irreversible Li+ consumption. From this perspective, a concise overview of diverse pre‐lithiation methodologies is provided and the reaction mechanisms associated with over‐ and hyper‐lithiated oxides as sacrificial cathode additives for pre‐lithiation are investigated. Subsequently, strategies to modulate the electrochemical properties of these oxides for practical use in sacrificial cathodes are briefly explored. Following this, discussions are carried out and perspectives on research that adopts the aforementioned contrasting directions are presented.

Optimized SMRT-UMI protocol produces highly accurate sequence datasets from diverse populations-Application to HIV-1 quasispecies.

Pathogen diversity resulting in quasispecies can enable persistence and adaptation to host defenses and therapies. However, accurate quasispecies characterization can be impeded by errors introduced during sample handling and sequencing, which can require extensive optimizations to overcome. We present complete laboratory and bioinformatics workflows to overcome many of these hurdles. The Pacific Biosciences single molecule real-time platform was used to sequence polymerase-chain reaction (PCR) amplicons derived from cDNA templates tagged with unique molecular identifiers (SMRT-UMI). Optimized laboratory protocols were developed through extensive testing of different sample preparation conditions to minimize between-template recombination during PCR. The use of UMI allowed accurate template quantitation as well as removal of point mutations introduced during PCR and sequencing to produce a highly accurate consensus sequence from each template. Production of highly accurate sequences from the large datasets produced from SMRT-UMI sequencing is facilitated by a novel bioinformatic pipeline, Probabilistic Offspring Resolver for Primer IDs (PORPIDpipeline). PORPIDpipeline automatically filters and parses circular consensus reads by sample, identifies and discards reads with UMIs likely created from PCR and sequencing errors, generates consensus sequences, checks for contamination within the dataset, and removes any sequence with evidence of PCR recombination, heteroduplex formation, or early cycle PCR errors. The optimized SMRT-UMI sequencing and PORPIDpipeline methods presented here represent a highly adaptable and established starting point for accurate sequencing of diverse pathogens. These methods are illustrated through characterization of human immunodeficiency virus quasispecies in a virus transmitter-recipient pair of individuals.

Why coastal lagoons are so productive? Physical bases of fishing productivity in coastal lagoons

Coastal lagoons are among the most productive marine ecosystems in the world. Annual primary production varies from 50 to > 500 g C m−2 year−1, being of the same order of magnitude as that of the upwelling areas. Many lagoons lie within the range of eutrophic (300–500 g C m−2 year−1) or hypereutrophic (> 500 g C m−2 year−1) conditions. The high productivity of coastal lagoons makes them subject of exploitation by many marine fishes and invertebrates, that use them as nursery areas and feeding grounds during their early life cycle phases, and most lagoons support important fisheries or maintain aquaculture exploitations. The high levels of their biological production can be explained by some of their common features as shallowness and the strong influence of terrestrial systems. Shallowness favors that the photic zone extends to the lagoon bottom and that wind can promote the resuspension of nutrients and organisms. The interaction with land also introduces significant amounts of nutrients. However, trophic variables can explain < 43 % of the fishing yields, and further than the trophic status of the lagoons, several works showed that the biological productivity of coastal lagoons can be explained by their geomorphological features such as the positive influence of shoreline development and the negative influence of depth. Using the Mar Menor lagoon as a case study, we propose that although nutrient inputs and light can be limiting factors for photosynthetic based productivity, increasing fishing yield up to a certain limit, the productivity of lagoons is mainly promoted by more general forces associated to physical and chemical gradients.

Telomerase inhibitors induce mitochondrial oxidation and DNA damage-dependent cell death rescued by Bcl-2/Bcl-xL

BackgroundReactivation of telomerase is a hallmark of cancer and the majority of cancers over-express telomerase. Telomerase-dependent telomere length maintenance confers immortality to cancer cells. However, telomere length-independent cell survival functions of telomerase also play a critical role in tumorigenesis. Multiple telomerase inhibitors have been developed as therapeutics and include anti-sense oligonucleotides, telomerase RNA component targeting agents, chemical inhibitors of telomerase, small molecule inhibitors of hTERT, and telomerase vaccine. In general, telomerase inhibitors affect cell proliferation and survival of cells depending on the telomere length reduction, culminating in replicative senescence or cell death by crisis. However, most telomerase inhibitors kill cancer cells prior to significant reduction in telomere length, suggesting telomere length independent role of telomerase in early telomere dysfunction-dependent cell death. MethodsIn this study, we explored the mechanism of cell death induced by three prominent telomerase inhibitors utilizing a series of genetically encoded sensor cells including redox and DNA damage sensor cells. ResultsWe report that telomerase inhibitors induce early cell cycle inhibition, followed by redox alterations at cytosol and mitochondria. Massive mitochondrial oxidation and DNA damage induce classical cell death involving mitochondrial transmembrane potential loss and mitochondrial permeabilization. Real-time imaging of the progression of mitochondrial oxidation revealed that treated cells undergo a biphasic mitochondrial redox alteration during telomerase inhibition, emphasizing the potential role of telomerase in the redox regulation at mitochondria. Additionally, silencing of hTERT confirmed its predominant role in maintaining mitochondrial redox homeostasis. Interestingly, the study also demonstrated that anti-apoptotic Bcl-2 family proteins still confer protection against cell death induced by telomerase inhibitors. ConclusionThe study demonstrates that redox alterations and DNA damage contribute to early cell death by telomerase inhibitors and anti-apoptotic Bcl-2 family proteins confer protection from cell death by their ability to safeguard mitochondria from oxidation damage.

Blastocyst telomere length predicts successful implantation after frozen-thawed embryo transfer.

Do embryos with longer telomere length (TL) at the blastocyst stage have a higher capacity to survive after frozen-thawed embryo transfer (FET)? Digitally estimated TL using low-pass whole genome sequencing (WGS) data from the preimplantation genetic testing for aneuploidy (PGT-A) process demonstrates that blastocyst TL is the most essential factor associated with likelihood of implantation. The lifetime TL is established in the early cleavage cycles following fertilization through a recombination-based lengthening mechanism and starts erosion beyond the blastocyst stage. In addition, a telomerase-mediated slow erosion of TL in human fetuses has been observed from a gestational age of 6-11 weeks. Finally, an abnormal shortening of telomeres is likely involved in embryo loss during early development. Blastocyst samples were obtained from patients who underwent PGT-A and FET in an IVF center from March 2015 to May 2018. Digitally estimated mitochondrial copy number (mtCN) and TL were used to study associations with the implantation potential of each embryo. In total, 965 blastocysts from 232 cycles (164 patients) were available to investigate the biological and clinical relevance of TL. A WGS-based workflow was applied to determine the ploidy of each embryo. Data from low-pass WGS-PGT-A were used to estimate the mtCN and TL for each embryo. Single-variant and multi-variant logistic regression, decision tree, and random forest models were applied to study various factors in association with the implantation potential of each embryo. Of the 965 blastocysts originally available, only 216 underwent FET. While mtCN from the transferred embryos is significantly associated with the ploidy call of each embryo, mtCN has no role in impacting IVF outcomes after an embryo transfer in these women. The results indicate that mtCN is a marker of embryo aneuploidy. On the other hand, digitally estimated TL is the most prominent univariant factor and showed a significant positive association with pregnancy outcomes (P < 0.01, odds ratio 79.1). We combined several maternal and embryo parameters to study the joint effects on successful implantation. The machine learning models, namely decision tree and random forest, were trained and yielded classification accuracy of 0.82 and 0.91, respectively. Taken together, these results support the vital role of TL in governing implantation potential, perhaps through the ability to control embryo survival after transfer. The small sample size limits our study as only 216 blastocysts were transferred. The number was further reduced to 153 blastocysts, where pregnancy outcomes could be accurately traced. The other limitation of this study is that all data were collected from a single IVF center. The uniform and controlled operation of IVF cycles in a single center may cause selection bias. We present novel findings to show that digitally estimated TL at the blastocyst stage is a predictor of pregnancy capacity after a FET cycle. As elective single-embryo transfer has become the mainstream direction in reproductive medicine, prioritizing embryos based on their implantation potential is crucial for clinical infertility treatment in order to reduce twin pregnancy rate and the time to pregnancy in an IVF center. The AI-powered, random forest prediction model established in this study thus provides a way to improve clinical practice and optimize the chances for people with fertility problems to achieve parenthood. This study was supported by a grant from the National Science and Technology Council, Taiwan (MOST 108-2321-B-006-013 -). There were no competing interests. N/A.