Abstract Introduction: Pancreatic ductal adenocarcinoma (PDAC) arises from distinct precursor lesions, mainly pancreatic intraepithelial neoplasias (PanINs) and intraductal papillary mucinous neoplasms (IPMNs), which differ in genetics, differentiation ecosystem, and clinical behavior. PanIN-derived PDAC typically demonstrate more molecular and spatial heterogeneity, with pronounced classical-basal divergence. IPMN-derived adenocarcinomas display different histologic types: intestinal-type lesions often develop colloid carcinoma, where adenocarcinomas arising from pancreatobiliary(PB)-type IPMN resemble conventional PanIN-derived PDAC. Defining how precursor origin shapes malignant states, spatial microenvironment ecosystems are essential for PDAC classification. Method: Single-cell spatial transcriptomics was performed by CosMx SMI (Bruker) in whole transcriptome and 6K-panel. Two different Tissue Microarrays (TMAs) cohorts were used (1.5-mm cores) include PanIN-origin PDAC (15 patients) and IPMN-origin PDAC (10 patients). Cell Segmentation performed by CellPose and strict quality control (QC) thresholding yielded 459,195 high-quality cells. Data was log-normalized and integrated with scVI using patient-level correction. Hierarchical annotation was performed by combining expert histopathological alignments with Leiden-based refinements. Downstream spatial analysis used Python based methods including Squidpy. Results: Cross-cohort integration revealed marked origin-dependent divergence in the malignant spatial atlas. Malignant epithelium in PanIN-PDAC exhibited higher heterogeneity with more diverse and dispersed cell states. In contrast, the tumor cells in IPMN-derived PDAC presented obvious differentiation features for each carcinoma type. Intestinal-type IPMN PDAC developed unique malignant program that exhibiting typical goblet-like subtype, which was not observed in either other IPMN-type PDAC or PanIN-derived PDAC. However, the PB-type IPMN-PDAC showed substantial similarity with PanIN-PDAC, sharing multiple malignant programs, primarily including pEMT, progenitor cell-like and ADM-like subtypes. Furthermore, PanIN-PDAC retained specific tumor programs as exocrine-like, squamous, and inflammatory-association basal-like. Notably, although PB-type in IPMN PDAC and PanIN-PDAC share similarities in some tumor states, further analysis revealed significant differences in the spatial patterns and cell communication between tumor and microenvironments (TME). This suggests potentially different evolution of microenvironments. Conclusion: Precursor origin fundamentally shapes PDAC malignant states and stromal-immune ecosystems. Together, this work establishes a spatial framework for understanding PDAC heterogeneity and highlights the importance of origin-aware stratification in clinic and biology. Citation Format: Tengyu Zhang, Assya Legrini, Mari-Claire McGuigan, Stephan B. Dreyer, Ghazal Latifi, Claire Kennedy Dietrich, Hannah Morgan, Eva Verkolf, Virginia Padoan, Yoana Doncheva, Joao Da Silva Filho, Michail Doukas, Bas Groot Koerkamp, Joanne Edwards, Nigel Jamieson. Spatially revealed malignant and microenvironment ecosystems in PanIN- and IPMN-derived PDAC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6199.

SMARCA4-deficient undifferentiated tumor (SMARCA4-UT) is a recently recognized and highly aggressive malignancy characterized by loss of SMARCA4 (BRG1) expression. Although initially described in thoracic locations, involvement of the esophagus is rare, and cytopathologic experience remains limited, particularly using liquid-based cytology (LBC). A 51-year-old man presented with cervical lymphadenopathy and widespread metastatic disease. Excisional lymph node biopsy established the diagnosis of SMARCA4-UT. After initial chemotherapy and immunotherapy, he developed progressive dysphagia, and imaging demonstrated upper-esophageal stenosis and mural thickening. Cytologic evaluation of both a cervical lymph node and a subcutaneous back lesion using LBC showed moderately to markedly cellular preparations composed of dispersed malignant cells and loosely cohesive clusters in a necrotic background. Tumor cells displayed marked nuclear pleomorphism, eccentrically located nuclei, prominent nucleoli, and focal rhabdoid features. Immunohistochemical studies showed focal synaptophysin positivity but absence of chromogranin, INSM1, epithelial, and lymphoid markers. SMARCB1 (INI1) expression was retained, whereas SMARCA4 (BRG1) showed markedly reduced to complete loss of nuclear expression, confirming SMARCA4-UT. Esophageal biopsy demonstrated identical findings, supporting esophageal involvement. Despite treatment, the disease progressed rapidly, and the patient died 9 months after diagnosis. This case demonstrates that LBC specimens from metastatic sites can preserve diagnostic cytomorphologic features of SMARCA4-UT and highlights this rare presentation with esophageal involvement and reproducible findings across two metastatic sites. Awareness of cytologic features and potential diagnostic pitfalls is essential for timely diagnosis and appropriate ancillary testing.

Abstract Tumor heterogeneity fueled by plasticity and genetic diversification of cancer cells is key to therapy failure of malignant glioma. We implemented spatial and genetic platforms at single cell resolution to explore the trajectory of evolution of glioblastoma. Using spatial analysis of whole glioblastoma sections, we established a homotypic clustered cell identity paradigm whereby tumor cell state coherence was maximal in cells organized in homotypic clusters, whereas dispersed cells downregulated the original state, acquired alternative phenotypes and exhibited changes in the microenvironment, thus linking the process of plasticity to loss of the cell adhesion mechanisms that preserve the clustered spatial pattern of glioblastoma cells. We also used single cell DNA-sequencing methods integrated with the single cell transcriptome of patient-matched primary-recurrent glioblastoma pairs to resolve the clonal substructure of untreated glioblastoma and determine the clonal evolution at recurrence driven by therapeutic resistance. The evolutionary trajectory of glioblastoma identified a bottleneck model as the predominant pattern of evolution and converged on the identification of a rare persister subclonal state in primary glioblastoma exhibiting distinct phenotypic hallmarks that evolves and diversifies to populate the recurrent tumor mass. We used preclinical tumor models to trace the individual lineages associated with the persister subclones and experimentally illuminated the biological and metabolic activities of the persister cellular state in brain tumors. Persister glioblastoma cells in untreated tumors lacked spatial segregation and were independent predictors of timing to recurrence for glioblastoma patients. Thus, genetic and non-genetic co-evolution mechanisms forge the acquisition of plasticity and therapy resistance in glioblastoma. Citation Format: Antonio Iavarone. Shaping tumor cell plasticity and therapy resistance in glioblastoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Brain Cancer; 2026 Mar 23-25; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2026;86(6_Suppl):Abstract nr IA005.

Background/Objectives: Bacterial biofilms are structured communities of sessile cells embedded in a self-produced extracellular matrix. Within biofilms, bacteria become highly tolerant toenvironmental stressors such as host immune responses and antimicrobial treatments. In response to specific cues, however, biofilm cells can revert to a planktonic free-swimming lifestyle through a process termed biofilm dispersal. When dispersed cells escape the biofilm matrix, they lose biofilm-associated antibiotic tolerance, a major barrier to treating medical biofilms. As such, dispersal-inducing compounds like nitric oxide (NO) are actively investigated as adjuvants to potentiate the biofilm-eradicating activity of existing antibiotics. We recently characterised the transcriptomic responses elicited during spontaneous biofilm dispersal in closed culture-grown Pseudomonas aeruginosa biofilms. Here, we evaluated the transcriptional profiles of P. aeruginosa biofilms treated with the NO donor Spermine-NONOate (SP-NONO) and the nitroxide C-TEMPO, an NO analogue, to determine potential pathways involved in NO-mediated dispersal. Methods: Dispersal activity on P. aeruginosa PAO1 biofilms by SP-NONOate and C-TEMPO was quantified by crystal violet staining. Cellular responses to each compound were profiled by RNA-seq on treated and untreated cells. Results: While both compounds disrupted the transcription of ANR-regulated energy metabolism pathways, only SP-NONO activated canonical NO-regulated responses. Considering that only SP-NONO showed biofilm dispersal activity in this culture system, we investigated shared transcriptional shifts in SP-NONO-treated and spontaneously dispersed biofilms to identify pathways likely involved in central dispersal responses. These mostly included genes involved in the catabolism of branched-chain amino acids (leucine, valine, isoleucine) and lysine, as well as 9 of 14 genes previously defined as transcriptional biomarkers of spontaneous biofilm dispersal. Conclusions: This study suggests that NO disrupts biofilm maturation by prematurely stimulating central pathways of spontaneous biofilm dispersal and highlights this set of biomarkers as robust indicators of dispersal responses.

IL-2 synergizes with proinflammatory type 3 inducers to amplify mixed type 2-type 3 inflammation in nasal polyps.

Osteosarcoma is a malignant bone tumor, which can develop with the formation of immature bone or osteoid. It most commonly affects in children and adolescents, while its occurrence in elderly patients is relatively rare. In elderly patients, osteosarcoma can arise with paget’s disease, a condition characterized by abnormal bone remodeling. This report presents an unusual case of osteosarcoma in elderly patients highlighting the importance of radiological and biopsy evaluation in establishing the diagnosis. A 66-year-old man presented with a history of a lump on the right lower extremity, present for the past ten years and progressively enlarging over the last six months. patient experienced pain both at rest and during movement. There was no history of trauma, falls, comorbidities or family history that were associated with disease. Plain radiographs, MRI, and needle biopsy were performed. Imaging studies demonstrated bone destruction, periosteal reaction, soft tissue extension, and matrix ossification which are characteristic findings of osteosarcoma. Pathological examination identified a clusters and dispersed anaplastic cells with round-to-oval, pleomorphic, and hyperchromatic nuclei. Incidence of osteosarcoma in elderly patients is rare and show a poorer prognosis than do children and adolescent patients. In this age group, osteosarcoma often arises as a secondary malignancy. Conditions such as Paget’s disease or prior malignancies are known contributors. Several predisposing factors for adult-onset osteosarcoma have been identified, including underlying genetic syndromes, benign bone lesions, and a history of radiation therapy or chemotherapy. Early diagnosis in older individuals is important often challenging due to atypical presentations, often leading to misdiagnosis or delayed recognition. In elderly patients, bone tumors require careful diagnostic workup. Plain radiography play central role in initial evaluation, MRI delineates soft tissue involvement and metastatic spread, and histopathology confirms the diagnosis.

As an immediate countermeasure against cyanobacterial blooms in aquatic systems, on-site pulverization of cyanobacterial slurry may stimulate trophic transfer by enhancing the utilization of cyanobacteria by zooplankton. We verified the efficiency and effects of pulverizing cyanobacteria using an impinging jet device in a river connected to Lake Taihu, China. Colonies of Microcystis (mainly M. aeruginosa), ranging in size from 100 to 2000μm in the surface scums, were dispersed into single cells smaller than 10μm by the treatment, which consequently reduced Microcystis cell density at the surface of the treated site. Nevertheless, intact cells were often observed, suggesting that the dissociated cells retained their vesicles/vacuoles and buoyancy, which are typically damaged by other techniques (e.g., ultrasound, hydraulic cavitation). Microcystins in the ambient water did not increase after the treatment, which also indicates that the cell damage and subsequent release of toxins were limited. A laboratory experiment using dispersed Microcystis cells revealed that their edibility increased for bottom-dwelling Ostracoda, whereas it decreased for the completely planktonic Daphnia magna, probably because the cells were damaged and deposited in the experiment using ultrasound. These results suggest that the impinging jet can pulverize cyanobacteria with minimal cell damage and increase their edibility for zooplankton, particularly when the cells are retained in the water column.

Metastatic lobular carcinoma of the breast in serous effusions is diagnostically challenging because the tumor cells simulate histiocytes and mesothelial cells. The typical metastatic pattern is isolated dispersed tumor cells which are unrelated to surrounding mesothelial cells. Metastatic deposits of lobular carcinoma cells within mesothelial cell clusters have not been previously reported. We report our findings of peritoneal metastatic lobular breast carcinoma cells masquerading in atypical hyperplastic mesothelial cell clusters found in cellblock sections, but were not present in ThinPrep slides. Three patients out of 10 patients diagnosed with breast lobular carcinoma showed infrequent large atypical cell clusters in cellblock sections. They were not identified in ThinPrep slides which showed chronic lymphohistiocytic inflammatory hemorrhagic effusion fluids, that initially were reported as negative for malignant cells and disregarded as mixed inflammatory infiltrates of lymphocytes, plasma cells and histiocytes. However, cellblock sections showed occasional large cell clusters that were confused with atypical mesothelial cell clusters and metastatic adenocarcinoma. Cellblock immunocytochemistry showed peculiar staining patterns. BerEP3 and CEA showed scattered single mononuclear epithelial cells intimately dispersed within WT1 and calretinin-positive mesothelial cell clusters. An extended panel showed intramesothelial epithelial tumor cells expressing GATA3, but were negative for TTF1, Napsin-A, PAX8, CDX2. The tumor cells also expressed ER and mammaglobin. They did not express E-cadherin. The cytologic diagnosis was metastatic lobular carcinoma of the breast. This was confirmed by breast core needle biopsies. Because this phenomenon is not always apparent in ThinPrep slides, cellblock sections supplemented with immunocytochemistry are a valuable diagnostic tool. The remaining seven patients showed the usual dispersed patterns of metastatic lobular carcinoma in ThinPrep slides and cellblock sections in pleural and peritoneal effusions.

Understanding how cells respond to internal and external inputs requires investigating cells within three-dimensional (3D) environments, which better mimic physiological conditions. Compared to two-dimensional (2D) systems, 3D cultures more accurately simulate tissue architecture, including cell-cell and cell-extracellular matrix interactions, as well as gradients of oxygen and nutrients. Despite these advantages, quantifying signaling dynamics in 3D remains difficult due to limitations in imaging depth, phototoxicity, and computational analysis. We developed experimental and computational tools for tracking individual cells' responses in 3D. We focused on the response of human breast cancer cells to irradiation using a cell line that expresses a fluorescent reporter for the cell cycle regulator p21, which is activated by the tumor suppressor p53 after irradiation. We embedded individual cells and multicellular spheroids in a dual-Matrigel assay and used light sheet fluorescence microscopy (LSFM) to obtain high-resolution images at several time points post-irradiation. We then developed computational pipelines to obtain detailed reconstructions and quantitative analyses of p21 dynamics. Individual dispersed cells exhibited a gradual, monotonic increase in the fraction of p21-positive cells, with the majority of cells becoming positive 24 h after irradiation. When applied to spheroids, the same system captured a transient decrease in the fraction of p21-positive cells post-irradiation, followed by a delayed pronounced rise only at 24 h. In addition, while the fraction of p21-positive cells increased in both systems, p21 intensity within induced cells remained relatively constant. This behavior is consistent with studies in 2D cultures showing that irradiation induces p53 oscillations, with each p53 pulse regulating the probability, rather than the magnitude, of p21 transcription. Notably, spatial mapping of annotated nuclei showed no dependence between p21 levels and radial cell position within spheroids. Comparisons between 2D, 3D single-cell, and spheroid data indicate that while the overall extent of p21 activation is similar across systems, the kinetics differ, with spheroids exhibiting slower induction. The differences in features such as p21 induction kinetics observed in spheroids compared to 2D and 3D single-cell cultures post-irradiation likely reflect p21 signaling specific to cells in 3D configurations with cell-extracellular matrix constraints. Overall, the platform developed in this study provides a powerful framework to dissect heterogeneous signaling dynamics in physiologically relevant 3D contexts and can be extended to assess the effects of drug treatments on other complex multicellular structures.

Intestinal helminth parasites' impact on the health of their hosts in several ways, including altering the structure and function of host tissues. Histological impacts of three helminthes from three vertebrate hosts are hereby reported. Intestinal helminth community of greater cane rats (Thryonomys swinderianus) of Omagwa Bush Meat Market, Rivers State, Nigeria, was sampled to compare with previous reports. Intestinal tracts of T. swinderianus (n=12) samples were obtained from the slaughterhouse at Omagwa Bush Meat Market. Fish specimens (Bostrychus africanus [n=24] and Periophthalmus papilio [n=17]) were obtained from fishers at Creek Road Market, Rivers State. Sampling lasted from June to August 2023. All hosts were examined for parasites by dissection and microscopic examination; histologic impacts were assessed using Hematoxylin and Eosin staining procedures. Hosts and parasites were appropriately identified using keys. Thryonomys swinderianus were infected by nematodes- Trichuris sp. (16.7%), Oesophagostomum venulosum (50%) and Strongylus sp. (25%). Raphidascaroides africanus (20.8%) and Neoechinorhynchus sp. (6%) infected Bostrychus africanus and Periophthalmus papilio, respectively. Histologic impacts observed were as follows: T. swinderianus infected with Trichuris sp. showed mild disintegration of tissues. Raphidascaroides africanus caused detachment of the connective tissue core and dispersed mucus-secreting cells in B. africanus; Neoechinorhynchus sp. resulted in loss of secretory cells associated with disruption of the intestinal epithelial lining in P. papilio. The study showed that parasites result in histological changes, including tissue disintegration and dispersal/loss of secretory cells, in vertebrate hosts, and helminths of T. swinderianus include Trichuris sp., Oesophagostomum venulosum and Strongylus sp.

This study aimed to explore roles of β-catenin, collagen&elastic fibers in progression of Oral Epithelial Dysplasia (OED) to oral squamous cell carcinoma (OSCC). Sixty paraffin wax blocks of oral mucosa were divided into: control (Group I), mild OED (Group II), severe OED (Group III), well differentiated SCC (Group IV), moderately differentiated SCC (Group V), poorly differentiated SCC (Group VI). The sections underwent staining using hematoxylin and eosin, histochemical stains and β-catenin immunostaining. Group I revealed normal gingival tissue. Group II showed some basal and prickle cells with pleomorphic nuclei. Group III illustrated poorly demarcated cell membrane. Group IV showed keratinized epithelial islands. Group V was devoid of keratin pearls. Group VI showed dispersed epithelial cells. β-catenin-immunoreactivity was; strong membranous in Group I, moderate membranous, cytoplasmic and nuclear in Groups II&III, moderate nuclear and cytoplasmic in Group IV but strong in Groups V&VI. Collagen fibers were closely packed in Groups I&II, loosely packed in Groups III&IV, dispersed in Groups V&VI. Elastic fibers in Group I were apparently few & thin, abundant in Group II, numerous in Group III, short in Groups IV&V and thick in Group VI. β-catenin and elastic fibers were upregulated meanwhile collagen formation was downregulated towards progression from mild OED to poorly differentiated SCC.

Biofilms represent intricate microbial consortia adhering to surfaces, posing substantial challenges in medical and industrial contexts due to their formidable resistance to antimicrobial agents and host immune responses. These communities exhibit enhanced resilience over planktonic cells, necessitating innovative control and eradication strategies. Inducing biofilm dispersion has emerged as a promising strategy, garnering significant research interest. This review synthesizes recent advancements in biofilm dispersion, delineating the characteristics of dispersed cells, elucidating the molecular and environmental triggers of dispersion, and examining the ramifications of dispersal events. Furthermore, it underscores the therapeutic potential of leveraging dispersion to manage biofilm-associated infections and mitigate biofilm formation. By dissecting the mechanisms and implications of biofilm dispersion, this review enriches our understanding of biofilm dynamics and offers novel avenues for addressing biofilm-related challenges across diverse environments. Ultimately, harnessing dispersal mechanisms presents a paradigm shift in biofilm control, offering a powerful strategy to overcome treatment failures in chronic infections and to enhance efficacy in industrial biofouling mitigation.

Abstract Introduction/Objective Extracavitary primary effusion lymphoma (EC-PEL) is a rare and aggressive HHV8 and EBV-associated B-cell lymphoma that presents as a solid tumor rather than a typical effusion. With few reported cases, it remains underrecognized. EC-PEL often affects immunocompromised individuals, particularly those with HIV. Methods/Case Report A 46-year-old HIV-positive male presented with abdominal pain, nausea, vomiting, and fever. Imaging revealed a large retroperitoneal mass with pancreatic invasion, encasement of vascular structures, and retroperitoneal lymphadenopathy. Fine needle aspiration of the mass showed numerous dispersed neoplastic cells with immunoblastic, plasmablastic, or anaplastic features. Immunostains showed that the neoplastic cells were positive for CD138, MUM-1, CD4, CD30, HHV-8 and EBER, while negative for CD3, CD5, CD20, PAX5, CD79a, ALK, Cyclin D1 and TdT. In situ hybridization studies demonstrated a lambda-restricted pattern, consistent with diagnosis of EC-PEL. Clonal B cell immunoglobulin kappa rearrangement was detected by PCR. Bone marrow biopsy was negative. The patient underwent chemotherapy initially, but his condition deteriorated and passed away two months after the diagnosis. Results NA Conclusion EC-PEL can pose a diagnostic challenge due to the absence of common B- and T-cell markers. Early recognition of this rare entity, along with appropriate tissue acquisition, is crucial. Adequate cytology specimens can lead to an accurate diagnosis.

Bacterial cell aggregation plays a fundamental role in surface colonization, stress tolerance, and interspecies metabolite exchange. Aggregation is assessed by simple tube-settling assays and also image analysis; however, approaches for quantitatively assessing the heterotypic and homotypic cell–cell interactions among more than two types of cells have been limited. In this study, we developed grid partitioning image analysis (GPIA), a simple workflow that quantifies the compositional heterogeneity of bacterial aggregates. Confocal laser scanning microscopy (CLSM) images of fluorescently labeled Acinetobacter sp. Tol 5, which exhibits a self-aggregative nature through its cell surface protein AtaA, were partitioned into 2-μm square grids. Grids containing one or no cells were classified as dispersed, whereas those containing multiple cells were classified as aggregates, and the proportion of EGFP-labeled cells within each grid was recorded. Reference images representing dispersed cells, homo-aggregates, and hetero-aggregates produced characteristic EGFP-ratio histograms that matched binomial predictions. When AtaA production in one cell type was decreased, the histogram changed from a symmetric unimodal histogram with the peak at 40–60% EGFP-ratio to a skewed distribution, indicating that GPIA can detect differences in cell-to-cell affinity. Using the same procedure, we examined six in-frame deletion variants of AtaA. The deletion of the N-terminal head domain alone prevented co-aggregation with full-length AtaA, suggesting that homophilic recognition by this domain mediated self-aggregation, whereas deletions in all other regions had no measurable effect. GPIA, therefore, offers a simple and rapid approach for quantitative studies on bacterial cell aggregation, bridging the gap between qualitative microscopy and quantitative but technically demanding single-cell analysis. GPIA will accelerate research on cell–cell interactions, which are the foundational processes that drive biofilm formation and the assembly of microbial consortia.

BackgroundA decellularized liver scaffold (DLS) is a three-dimensional acellular extracellular matrix created by removing cellular components from liver tissue. Hepatocellular carcinoma (HCC) organoids represent a useful experimental model.MethodsHCC organoids from patient-derived xenografts (PDX), liver organoids, and HepG2 cells were expanded by cultivation within a murine DLS.ResultsHCC and liver organoids were generated from HCC PDX and human liver tissues, respectively. Expression levels of hepatocyte paraffin 1 (HepPar1), epithelial cell adhesion molecule (EpCAM), alpha-fetoprotein, keratin-7, and keratin-19 were detected in normal liver tissue, HCC tissue, HCC PDX, and HCC organoids. Fifteen murine DLSs were created, and the complete absence of liver cells was confirmed histologically by Masson trichrome and periodic acid–Schiff staining. Culture of HCC organoids in the DLS resulted in the expansion of numerous HCC cells scattered throughout the DLS framework. Keratin-7 expression was abundant, indicating widely dispersed progenitor cells. In contrast, cultivation of HepG2 cells within the DLS resulted in sparse cell distribution throughout the scaffold. Human hepatocyte organoids could not be cultivated successfully within the murine DLS framework.ConclusionsThe DLS collagen framework facilitates the proliferation of HCC organoids. Thus, cultivating HCC organoids within a DLS appears to represent an effective method for rapid cell expansion, although it was ineffective for HepG2 cells and liver organoids. Further studies are required to validate the biological and oncological equivalence between seeded HCC organoids and those rapidly expanded within a DLS.

Cardiomyocyte proliferative maturation and metabolic maturation occur in the same perinatal period. Intrauterine conditions influence developmental trajectories of both processes; however, their interconnectedness is unknown. Circulating fetal lipid levels are typically low, but the heart may be prematurely exposed to elevated lipids. We experimentally increased fetal lipid levels to determine the impact on cardiomyocyte proliferative maturation. Fetal sheep were surgically instrumented with catheters. After recovery, Intralipid 20 or Lactated Ringer's Solution were infused according to a clinical g/kg schedule for 8 days until 133 ± 1 days of gestation. A left ventricular biopsy was fixed, and remaining cardiomyocytes were enzymatically dissociated. Myocardial composition was measured from Masson's Trichrome-stained sections. Cardiomyocyte length, width, nucleation, and Ki-67+ were studied in dispersed cells. Myocardial composition, cardiomyocyte dimensions, and Ki-67+ were not found to be different between groups. Cardiomyocytes of the Intralipid-treated fetuses were 14% more terminally differentiated than controls (p = 0.025). Early developmental exposure to circulating lipid accelerates fetal cardiomyocyte terminal differentiation and may contribute to fewer cardiomyocytes for life.

To analyze the cytologic characteristics fine-needle aspiration using histology as the gold standard and to evaluate its diagnostic application in classic Hodgkin lymphoma. A retrospective analysis was conducted on 17 patients who underwent both coarse-needle aspiration and fine-needle aspiration and were histologically confirmed with classic Hodgkin lymphoma(CHL) at our hospital from December 2012 to December 2023. Clinical information of these patients was collected, and the smear morphology, immunocytochemistry and corresponding biopsies were reviewed. Among the 17 cases of CHL, there were 5 cases of mixed cellularity, 10 cases of nodular sclerosis and 2 cases were unsubtyped. Fifteen cases were correctly diagnosed by fine-needle aspiration, with an accuracy rate of 88.2%. The other two cases were misdiagnosed as non-Hodgkin lymphoma. Morphologically single dispersed mononuclear Hodgkin cells and multinucleated Reed-Sternberg cells were observed in a heterogenous background of lymphocytes in cytology smears, and these cells were positive for CD30 immunocytochemistry. Fine needle aspiration is less invasive and quicker, and the cell morphology is better preserved as compared to histological biopsy. It is easier to recognize pathognomonic Hodgkin or Reed-Sternberg cells and it is helpful for the rapid diagnosis and clinical management of CHL.

Introduction and Objective: Using a sensitive and specific mitochondrial membrane potential (ΔψM) assay, we found previously that mitochondrial energetics explains most of the GSIS in non-diabetic human islet cell cultures. Using T2D and non-diabetic β-cells in dispersed islet cell cultures and in intact islets, we defined a signature of T2D in β-cell bioenergetics, the response of ΔψM to oligomycin (an ATP-synthase inhibitor) in high glucose. The magnitude of this signature correlated with HbA1c and was recapitulated by culturing normal β-cells in high glucose. Using control analysis, we defined this signature as a metabolic activation mechanism that enables β-cell mitochondria to increase respiration while ATP/ADP is increasing. Methods: Here we have examined the mechanism of the activation of energy metabolism during GSIS, using dispersed cultures of human islet cells and fluorescence microscopy for analysis of ΔψM, NAD(P)H autofluorescence and succinate dehydrogenase (SDH) activity, and insulin assaying. β-cell-specific gene expression was determined by 10xGenomics single nuclei RNA sequencing. Results: We found that the activation of the energy metabolism during GSIS was an ATP-linked (or oligomycin-sensitive) increase in the activity of the TCA cycle. SDH can undergo ATP-linked activation, and here we found that its activity in human primary pancreatic β-cells increases in high glucose and is inhibited by deenergization by oligomycin plus FCCP (an uncoupler). Correlation analysis in a cohort of human islet donors revealed links between SDH activity and metabolic activation. SDH is inhibited by oxaloacetate, therefore we tested the effects of inhibiting the removal of matrix oxaloacetate by the phosphoenolpyruvate (PEP) carboxykinase reaction. CRISPR knockout of the PEP-cycle enzyme PCK2 proportionally decreased the ΔψM response and insulin secretion for glucose, but it did not alter the SDH activity or its sensitivity to deenergization. Conclusion: Altogether, the regulation of SDH activity is a possible novel component of GSIS. Disclosure C.A. Lerner: None. H. Cheng: None. A.A. Gerencser: None. Funding 1R01DK135807

The emerging Candidozyma auris (formerly known as Candida auris, C. auris) has caused several outbreaks globally. While several studies explored the resistant biofilm formed by C. auris, little is known regarding the cells dispersed following biofilm formation. Herein, I investigated and characterized the cells dispersed from C. auris biofilms. Cells dispersed from biofilm developed in 96 well plate were isolated and counted. The antifungal susceptibility testing showed that the dispersed cells display similar antifungal susceptibility as the parent planktonic cells, except amphotericin B. Gene expression analysis performed by quantitative real-time PCR indicated that dispersed cells can express genes coded for antifungal resistance (ERG2, ERG6, ERG11, FKS1, CHS1, CHS2, CDR1, MDR1) more than the parent planktonic cells. It was observed that dispersed cells can acquire resistance to caspofungin faster than the parent planktonic cells once exposed to caspofungin at sub MIC level. Furthermore, biofilms formed by dispersed cells displayed significantly higher metabolic activity, as indicated by the XTT analysis. To provide more insight, I explored the expression of genes coding for biofilm initiation and maturation and the data obtained indicated that dispersed cells overexpressed ALS5 and KRE6 genes. Further, GC-MS analysis indicated that dispersed cells exhibit altered metabolic profile that enhance cells survivability under stress and nutrient limit condition. The presented study is the first to explore C. auris dispersed cells and indicated that they are not able to revert to the planktonic mode once released from the biofilm.

Radiotherapy is the cornerstone of treatment for cervical cancer, yet the variability of patient response demands a deeper understanding of the molecular determinants of radioresistance. In this study, we investigated the molecular and cellular mechanisms of radioresistance in cervical cancer through a comprehensive multi-omics and machine learning approach. We downloaded and processed transcriptome sequencing, methylation and single-cell sequencing data from the TCGA and GEO databases. Differential gene and methylation analyses were performed to identify radioresistance-related markers. Single-cell data were processed using Seurat and annotated using CellTypist. Prognostic models were constructed and validated through downscaling, cell scoring, trajectory analysis and machine learning. Additionally, immune infiltration and drug sensitivity analyses were conducted. The differential analysis identified 845 up-regulated and 460 down-regulated genes associated with radioresistance. The methylation analysis identified 3042 down-regulated and 158 up-regulated gene loci. Single-cell sequencing revealed 43,475 cells and 13 cell types, with aneuploid cells predominantly present in epithelial cells. Cell scoring highlighted dispersed immune cells, with monocytes, ILCs, and T cells being the most relevant to radiotherapy resistance. The machine learning approach constructed a robust prognostic model using Cox regression and validated it on multiple datasets. The prognostic model demonstrated good predictive ability in assessing radiotherapy efficacy and immune infiltration. Drug screening identified several potential therapeutic candidates with high sensitivity for high-risk patients. This study provides a comprehensive multi-omics analysis and machine learning framework for identifying and validating molecular markers and prognostic models associated with radioresistance in cervical cancer, providing insights for personalized treatment strategies.