Pattern Of Proliferation Research Articles

Proliferation Patterns of MCF-7 Breast Cancer Cells in Lipoaspirate Conditioned Media.

Autologous fat grafting (AFG) is a common technique used to enhance aesthetic outcomes in postmastectomy breast reconstruction patients. Adipokines are hormones secreted by adipose tissue that play a critical role in regulating metabolic processes and the immune system. However, dysregulated adipokine secretion and signaling can contribute to the development and progression of cancer by promoting angiogenesis, altering the immune response, and inducing the epithelial mesenchymal transition. We aimed to assess how breast cancer cells behave in conditioned media derived from fat grafting lipoaspirates and gain a better understanding of the potential interactions that may occur within the tumor microenvironment. Patients who were undergoing AFG as a part of breast reconstruction at NY-Presbyterian/Weill Cornell Medical Center between March 2021 and July 2023 were consented and enrolled in the study. This study was approved by the Weill Cornell Medicine Institutional Review Board (#20-10022850-14). Conditioned media is created using 20% of patient lipoaspirate secretome and 80% starving media. The growth of MCF-7, a human ER/PR+ breast cancer cell line, in conditioned media is assessed using CyQUANT. The breast cancer cells incubated in conditioned media displayed similar growth trends as those in complete media, which is enriched for cell growth (P > 0.05). MCF-7 cell behavior in conditioned media differed significantly from their proliferation patterns when serum starved in 100% starving media (P < 0.05). Our results suggest that there may be inherent factors within the lipoaspirate that may promote MCF-7 proliferation. One potential implication is that AFG used for breast reconstruction should be delayed until local-regional disease control has been established. In addition, based on the in vitro proliferation patterns of breast cancer cells in conditioned media, the safety profile of AFG may be enhanced if the procedure is performed after attaining negative margins and the completion breast cancer treatment.

A Comprehensive Approach: Correlating Ultrasound Imaging with Endometrial Histopathological Analysis in Perimenopausal Women with Heavy MenstrualBleeding.

Abnormal uterine bleeding (AUB) is a common troublesome symptom in the perimenopausal age group. The most common type of AUB in this age group is heavy menstrual bleeding. There is a risk of endometrial carcinoma and atypical endometrial hyperplasia in women with AUB in the age group of 40-50 years. Hence early evaluation is of paramount importance in managing women with perimenopausal heavy menstrual bleeding. The current study was undertaken to study the correlation between ultrasound findings and various benign and malignant endometrial histologies in perimenopausal women with heavy menstrual bleeding. Women aged 40-55 years presenting with heavy menstrual bleeding at the gynaecology outpatient department at Sree Balaji Medical College and Hospital, Chennai, India, were included in the study. Patients on anti-platelet and anti-coagulation therapy and patients already on hormonal treatment for heavy menstrual bleeding were excluded from the study. The demographic factors, symptom profiles, ultrasound findings, and histopathological reports were tabulated and analysed. Of the 147 women included in the study, 75 (51%) were aged 45-50 years and 107 (73%) had two or more pregnancies. Fibroid was the common non-endometrial cause of heavy menstrual bleeding in 52 (35%) cases. The proliferative pattern was the most common non-pathological histology identified in 46 (31%) cases. Endometrial hyperplasia without atypia was the most common pathological histology observed in the study population. Endometrial thickness of more than 8 mm was strongly associated with premalignant or malignant endometrial lesions. Our study has attempted to identify the correlation between ultrasound evaluation of perimenopausal women with heavy menstrual bleeding and endometrial pathology. Ultrasound, being cost-effective and widely available, is proven to be a tool for first-line investigation of perimenopausal women with heavy menstrual bleeding that guides further evaluation and management.

Endothelial cell proliferation and vascular patterns in urothelial carcinoma.

The bladder cancer has some characteristics: the sixth most incident neoplasm in the United States, the majority of diagnosed cases in those 55 years of age and older, four times more common in man than women, a reduced five-year survival rate in case of metastatic disease. Despite the beneficial effects of the combination therapy and immunotherapy, the low response rate and drug resistance were reported. The main goal of this work was evaluation of the endothelial cell proliferation from urothelial carcinomas. Fifty-two cases of T2-T4 infiltrative bladder tumors, aged between 46 and 78 years, were investigated. Morphological, simple and cluster of differentiation 31 (CD31)∕Ki67, CD31∕smooth muscle actin (SMA) double immunostaining were performed. In all the analyzed infiltrative bladder tumors, three types of vessels were noticed: immature, intermediate and mature. In the central part of the tumor area, the following distribution of vessel types was noticed: immature (62.25%), intermediate (35.1%), and mature vessels (2.65%). In the peripheral tumor area, the intermediate vessels increase numerically, up to 54% and the mature ones, up to 18.6%. The peritumoral area was characterized by the absence of immature vessels and the presence of intermediate and mature ones only. It was found the presence of endothelial cell nuclei stained for Ki67 only for immature and intermediate vessels, and never for mature ones. The vascular patterns may contribute to a better stratification of the patient subgroups and antiangiogenic treatment algorithms.

Histological criteria for selecting patients who need clonality test for non-gastric MALT lymphoma diagnosis

The histological diagnosis of extranodal marginal zone lymphoma of the mucosa-associated lymphoid tissue (MALT lymphoma) is difficult for pathologists. Recently, digital pathology systems have been widely used to provide tools that can objectively measure lesions on slides. In this study, we measured the extent of marginal zone expansion in suspected MALT lymphoma cases and compared the results with those of a molecular clonality test. In total, 115 patients who underwent an IGH gene rearrangement test for suspected MALT lymphoma were included in this study. All cases were histologically classified into three patterns; “small lymphoid aggregates with no germinal center (Pattern 1),” “lymphoid follicles with germinal center (Pattern 2)” and " fused marginal zone or diffuse small lymphocytic proliferation (Pattern 3).” The proportions of monoclonality in Pattern 1, 2, and 3 were 25.0%, 55.0%, and 97.9%, respectively. The ratios of marginal zone thickness to germinal center diameter and entire lymphoid follicle area to germinal center area were measured in Pattern 2 cases using a digital pathology system. Combining the width cutoff of 1.5 and the areal cutoff of 3.5, the sensitivity, specificity, positive predictive value, and negative predictive value for MALT lymphoma were 96.97%, 70.37%, 80.00%, and 95.00%, respectively. In conclusion, through objective measurement of the marginal zone, suspected cases of MALT lymphoma requiring a molecular clonality test can be effectively selected.

Mechanical control of cell proliferation patterns in growing epithelial monolayers

Cell proliferation plays a crucial role in regulating tissue homeostasis and development. However, our understanding of how cell proliferation is controlled in densely packed tissues is limited. Here we develop a computational framework to predict the patterns of cell proliferation in growing epithelial tissues, connecting single-cell behaviors and cell-cell interactions to tissue-level growth. Our model incorporates probabilistic rules governing cell growth, division, and elimination, also taking into account their feedback with tissue mechanics. In particular, cell growth is suppressed and apoptosis is enhanced in regions of high cell density. With these rules and model parameters calibrated using experimental data for epithelial monolayers, we predict how tissue confinement influences cell size and proliferation dynamics and how single-cell physical properties influence the spatiotemporal patterns of tissue growth. In this model, mechanical feedback between tissue confinement and cell growth leads to enhanced cell proliferation at tissue boundaries, whereas cell growth in the bulk is arrested, recapitulating experimental observations in epithelial tissues. By tuning cellular elasticity and contact inhibition of proliferation we can regulate the emergent patterns of cell proliferation, ranging from uniform growth at low contact inhibition to localized growth at higher contact inhibition. We show that the cell size threshold at G1/S transition governs the homeostatic cell density and tissue turnover rate, whereas the mechanical state of the tissue governs the dynamics of tissue growth. In particular, we find that the cellular parameters affecting tissue pressure play a significant role in determining the overall growth rate. Our computational study thus underscores the impact of cell mechanical properties on the spatiotemporal patterns of cell proliferation in growing epithelial tissues.

Multi-objective liver cancer algorithm: A novel algorithm for solving engineering design problems

This research introduces the Multi-Objective Liver Cancer Algorithm (MOLCA), a novel approach inspired by the growth and proliferation patterns of liver tumors. MOLCA emulates the evolutionary tendencies of liver tumors, leveraging their expansion dynamics as a model for solving multi-objective optimization problems in engineering design. The algorithm uniquely combines genetic operators with the Random Opposition-Based Learning (ROBL) strategy, optimizing both local and global search capabilities. Further enhancement is achieved through the integration of elitist non-dominated sorting (NDS), information feedback mechanism (IFM) and Crowding Distance (CD) selection method, which collectively aim to efficiently identify the Pareto optimal front. The performance of MOLCA is rigorously assessed using a comprehensive set of standard multi-objective test benchmarks, including ZDT, DTLZ and various Constraint (CONSTR, TNK, SRN, BNH, OSY and KITA) and real-world engineering design problems like Brushless DC wheel motor, Safety isolating transformer, Helical spring, Two-bar truss and Welded beam. Its efficacy is benchmarked against prominent algorithms such as the non-dominated sorting grey wolf optimizer (NSGWO), multiobjective multi-verse optimization (MOMVO), non-dominated sorting genetic algorithm (NSGA-II), decomposition-based multiobjective evolutionary algorithm (MOEA/D) and multiobjective marine predator algorithm (MOMPA). Quantitative analysis is conducted using GD, IGD, SP, SD, HV and RT metrics to represent convergence and distribution, while qualitative aspects are presented through graphical representations of the Pareto fronts. The MOLCA source code is available at: https://github.com/kanak02/MOLCA.

Histopathological spectrum of endometrial biopsies in abnormal uterine bleeding at tertiary care centre, Jodhpur

Introduction: Abnormal uterine bleeding (AUB) shows spectrum of patterns on histopathology and pathologist plays a vital role in reporting of endometrial biopsies and helps in differentiating nonneoplastic lesion from neoplastic lesions, early detection of the precursor lesions and exclusion of malignancy. Aim - The aim of this study is to study the spectrum of endometrial patterns in women with AUB and to correlate it with the different age groups. Materials and Methods: 200 samples of endometrial biopsies with AUB were received from the gynecology department, processed and stained with hematoxylin &amp; eosin and subjected to histopathological examination. Results: The peak incidence is observed in the age group of 41-50 years. The predominant histological pattern observed is secretory and proliferative pattern and disordered endometrium (68%) followed by endometrial hyperplasia (10%) and the least common pattern is atrophic endometrium (1%). 2% cases are of endometrial carcinoma found in our study. Conclusion: AUB significantly affects the quality life of women and leads to anemia. Hence histopathological examination should be considered which plays a critical role in early diagnosis of endometrial pathology and to provide appropriate gynecological management. Endometrial sampling is an effective diagnostic tool and it needs to be considered in all patients from peri- and post-menopausal age groups with AUB so to provide appropriate management.

Patterns of proliferation and fibrosis in a rat model of endometriosis following administration of Allium cepa.

Endometriosis is a common gynecological disease among reproductive-age women. Numerous hypotheses exist regarding the pathogenesis of endometriosis. In Turkey, the consumption of Allium cepa (commonly known as the "onion cure") is a popular treatment employed to alleviate a variety of gynecological disorders. In this study, our objective was to assess the therapeutic mechanisms of the onion bulb A. cepa using an autologous endometriosis model in Sprague-Dawley rats. Previous research has shown that A. cepa possesses anti-inflammatory, antioxidant, and antiapoptotic properties. We evaluated the pathological condition of endometriotic implants by employing hematoxylin-eosin staining and Ki67 immunohistochemistry analysis. Transforming growth factor-beta 1 (TGF-β1) and alpha-smooth muscle actin (α-SMA) have been identified as profibrotic markers that are highly overexpressed in endometriotic tissues relative to eutopic endometrial tissue. Furthermore, TGF-β1 influences the differentiation and progression of endometriosis. To quantify profibrotic activity, we measured TGF-β1 and α-SMA using the immunosorbent assay method. Lower histologic evaluation scores for endometriotic implants were observed in the group receiving high-dose A. cepa relative to the other groups. Ki67 expression was reduced following the high-dose A. cepa regimen, which consisted of 30% A. cepa and 70% normal feed. However, no statistically significant differences in TGF-β1 or α-SMA levels were observed among the groups (p=0.7 and p=0.778, respectively). The findings suggest that A. cepa could serve as a therapeutic agent in endometriosis treatment, as evidenced by the reduction in proliferative potential. Nevertheless, A. cepa was not associated with significantly lower levels of endometriosis-associated TGF-β1 or α-SMA.

Atypical Anti-Glomerular Basement Membrane Nephritis: A Case Series From the French Nephropathology Group

Rationale & ObjectiveAtypical anti-glomerular basement membrane (GBM) nephritis is characterized by a bright linear immunoglobulin staining along the GBM by immunofluorescence without a diffuse crescentic glomerulonephritis nor serum anti-GBM antibodies by conventional ELISA. We characterized a series of patients with atypical anti-GBM disease. Study DesignCase series. Setting & ParticipantsPatients were identified by the French Nephropathology Group as having atypical anti-GBM nephritis between 2003 and 2022. FindingsAmong 38 potential cases, 25 were included. 14 (56%) were female and 23 (92%) had hematuria. Median [interquartile range (IQR)] serum creatinine at diagnosis was 150 [102-203] μmol/L and median [IQR] urine protein to creatinine ratio was 2.4 [1.3-5.2] g/g. 9 (36%) patients had endocapillary proliferative glomerulonephritis (GN), 4 (16%) had mesangial proliferative GN, 4 (16%) had membranoproliferative GN, 2 (8%) had pure and focal crescentic GN, 1 (4%) had focal segmental glomerulosclerosis, and 5 had glomeruli that were unremarkable on histopathology. Nine patients (36%) had crescents, involving a median of 9% of glomeruli. Bright linear staining for IgG was seen in 22 cases (88%) and for IgA in 3 cases (12%). The nine patients (38%) who had a monotypic staining pattern tended to be older with less proteinuria and rarely had crescents. Kidney survival rate at one year was 83% and did not appear to be associated with the light chain restriction. LimitationsRetrospective case series with a limited number of biopsies including electron microscopy. ConclusionsCompared to typical anti-GBM disease, atypical anti-GBM nephritis frequently presents with endocapillary or mesangial proliferative glomerulonephritis pattern and appears to have slower disease progression. Further studies are needed to fully characterize its pathophysiology and associated clinical outcomes.

Seasonal mouse cadaver microbial study: rupture time and postmortem interval estimation model construction.

The estimation of postmortem interval (PMI) has long been a focal point in the field of forensic science. Following the death of an organism, microorganisms exhibit a clock-like proliferation pattern during the course of cadaver decomposition, forming the foundation for utilizing microbiology in PMI estimation. The establishment of PMI estimation models based on datasets from different seasons is of great practical significance. In this experiment, we conducted microbiota sequencing and analysis on gravesoil and mouse intestinal contents collected during both the winter and summer seasons and constructed a PMI estimation model using the Random Forest algorithm. The results showed that the MAE of the gut microbiota model in summer was 0.47±0.26 d, R2 = 0.991, and the MAE of the gravesoil model in winter was 1.04 ± 0.22d, R2 = 0.998. We propose that, in practical applications, it is advantageous to selectively build PMI estimation models based on seasonal variations. Additionally, through a combination of morphological observations, gravesoil microbiota sequencing results, and soil physicochemical data, we identified the time of cadaveric rupture for mouse cadavers, occurring at around days 24-27 in winter and days 6-9 in summer. This study not only confirms previous research findings but also introduces novel insights, contributing to the foundational knowledge necessary to advance the utilization of microbiota for PMI estimation.

Identification of novel T cell proliferation patterns, potential biomarkers and therapeutic drugs in colorectal cancer.

Background: T cells are crucial components of antitumor immunity. A list of genes associated with T cell proliferation was recently identified; however, the impact of T cell proliferation-related genes (TRGs) on the prognosis and therapeutic responses of patients with colorectal cancer (CRC) remains unclear. Methods: 33 TRG expression information and clinical information of patients with CRC gathered from multiple datasets were subjected to bioinformatic analysis. Consensus clustering was used to determine the molecular subtypes associated with T cell proliferation. Utilizing the Lasso-Cox regression, a predictive signature was created and verified in external cohorts. A tumor immune environment analysis was conducted, and potential biomarkers and therapeutic drugs were identified and confirmed via in vitro and in vivo studies. Results: CRC patients were separated into two TRG clusters, and differentially expressed genes (DEGs) were identified. Patient information was divided into three different gene clusters, and the determined molecular subtypes were linked to patient survival, immune cells, and immune functions. Prognosis-associated DEGs in the three gene clusters were used to evaluate the risk score, and a predictive signature was developed. The ability of the risk score to predict patient survival and treatment response has been successfully validated using multiple datasets. To discover more possible biomarkers for CRC, the weighted gene co-expression network analysis algorithm was utilized to screen key TRG variations between groups with high- and low-risk. CDK1, BATF, IL1RN, and ITM2A were screened out as key TRGs, and the expression of key TRGs was confirmed using real-time reverse transcription polymerase chain reaction. According to the key TRGs, 7,8-benzoflavone was identified as the most significant drug molecule, and MTT, colony formation, wound healing, transwell assays, and in vivo experiments indicated that 7,8-benzoflavone significantly suppressed the proliferation and migration of CRC cells. Conclusion: T cell proliferation-based molecular subtypes and predictive signatures can be utilized to anticipate patient results, immunological landscape, and treatment response in CRC. Novel biomarker candidates and potential therapeutic drugs for CRC were identified and verified using in vitro and in vivo tests.

Host-parasite interactions: a study on the pathogenicity of different Mesanophrys sp. densities and hemocytes-mediated parasitic resistance of swimming crabs (Portunus trituberculatus).

Mesanophrys sp. is a parasitic ciliate that invades and destroys the hemocytes of the swimming crab (Portunus trituberculatus). In the present study, we employed an in vitro model to elucidate how Mesanophrys sp. destroys crab hemocytes. We also evaluated the relationship between the parasite's density, the destruction rate of the hemocytes, and the rapid proliferation pattern of parasites in host crabs. We found that the survival rate and cell integrity of crab hemocytes decreased with an increase in Mesanophrys sp. density, depicting a negative correlation between hemocyte viability and parasite density. Further analyses revealed that crab hemocytes could resist destruction by a low density (10 ind/mL) of Mesanophrys sp. for a long time (60h). Mesanophrys sp. and its culture medium (containing the ciliate secretions) destroy the host hemocytes. The natural population growth rate of Mesanophrys sp. decreased with an increase in the parasite density, but the Mesanophrys sp. density did not affect the generation time of the parasites. In summary, Mesanophrys sp. can destroy crab hemocytes, and the degree of destruction is directly proportional to the parasite density. The resistance of crab hemocytes to Mesanophrys sp. decreased gradually with an increase in the parasite density.

Abstract B045: EV-mediated immune modulation of CD8+ T cells in renal cell carcinoma

Abstract Introduction: Treatment regimens for advanced renal cell carcinoma (RCC) involve immune checkpoint inhibition targeting the PD-(L)1 pathway. PD-L1 is often overexpressed on tumor cells in advanced disease and plays a crucial role in regulating tumor-directed immune responses. It can also be expressed on tumor-secreted extracellular vesicles (EVs), which are an essential part of intercellular communication. PD-L1 carrying EVs have been shown to have immunomodulatory effects in several tumor entities, however their role in RCC is still unknown. The aim of this study was the evaluation of putative immunomodulatory effects of RCC-derived small EVs on CD8+ T cells. Materials and Methods: EVs of 5 untreated as well as interferon γ (IFNγ)-treated RCC cell lines were isolated through serial ultracentrifugation. Quality and quantity of the EVs were verified using Western Blotting (WB), Nano Particle Tracking Analysis and Transmission Electron microscopy, their PD-L1 expression was analyzed semi quantitatively through WB. CD8+ T cells were isolated from the blood of healthy donors, activated and subsequently co-cultured with RCC-derived EVs for a duration of 3 days. Proliferation and PD-1 expression of the T cells was assessed using flow cytometry. ELISAs were performed to analyze cyto- and chemokine secretion. Results: All RCC cell lines expressed PD-L1, albeit at varying low levels. Four out of 5 treated cell lines responded to IFNγ treatment with an upregulation of PD-L1 expression on both cells and secreted EVs. Co-cultures of CD8+ T cells and RCC-derived EVs, particularly PD-L1+ EVs isolated from IFNγ-stimulated cells, lead to a reduction in T cell PD-1 expression. Additionally, PD-L1-expressing EVs of RCC cell line RCC53 had an antiproliferative effect on CD8+ T cells. EVs also increased the secretion of IL-2, IL-10, CXCL10 and CCL5 by the CD8+ T cells. Conclusions: PD-L1 is expressed at low levels in RCC cell lines. However, repetitive IFNγ stimulation can lead to upregulation of PD-L1 on both cells and EVs enabling us to use this model to further investigate EV-mediated, PD-L1 dependent effects. In co-cultures, RCC-derived EVs impede early T cell activation and proliferation patterns of CD8+ T cells. Furthermore, early results indicate a shift towards a more pro-inflammatory cyto- and chemokine production by the T cells induced by RCC-derived EVs. The results support the hypothesis of a tumor-EV-mediated immune modulation in RCC. Citation Format: Greta Jaschkowitz, Elfriede Noessner, Angela Zaccagnino, Michael Stöckle, Kerstin Junker, Philip Zeuschner. EV-mediated immune modulation of CD8+ T cells in renal cell carcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B045.

Developing Deep Learning Pipeline of Whole-Slide Images for Enhanced Diffuse Large B Cell Lymphoma (DLBCL) Subtyping and Outcome Prediction: Leveraging Self-Attention Transformer for Training and Inference

Background: Deep learning algorithms can help to analyze whole-slide images (WSI) in lymphoma pathology, identifying deeper features and patterns that may not be easily discernible to human observers. This pilot project is focused on diffuse large B-cell lymphoma (DLBCL), a heterogeneous disease with diverse genetic alterations. By leveraging self-attention trained clusters and transformers, the project aims to identify patterns and associations between mutational status and overall survival, potentially enhancing personalized treatment strategies and improving data reliability. Methods: This study employed a computer vision pipeline learning system to classify DLBCL subtypes using self-discovery of discriminatory features from scanned WSI. The workflow is shown in Figure 1a. First, we segmented tiles or patches from the gigapixel-sized WSI of 223 lymphoma cancer biopsy slides sourced from The Cancer Genome Atlas (TCGA), DLBCL, and Stanford DLBCL-Morph datasets. to optimize extraction of relevant features. For feature extraction, we utilized self-supervised pretraining with a Vision Transformer (ViT) network on a dataset of 1,515,000 patches. These patches were grouped into morphologically similar clusters using K-means, representing various lymphoma proliferation patterns. Dimensionality reduction with UMAP allowed for computational efficiency and feature visualization. Extracted features were used to predict overall survival using Bag-of-Words (BoWs). We further enhanced the model by incorporating geometric information, including nuclear characteristics from HoverNet. We assessed the model's performance using key metrics such as area under the curve (AUC) and accuracy. Additionally, genomic mutations with a frequency of 10% or higher from the TCGA were incorporated for enhanced predictive capability, including PIM1, SGK1, CARD11, KMT2D, SOCS1, BTG1, MUC16 and FAT4. Correlations and hierarchical clustering were performed on mutations, patches, and outcomes (Figure 1b). Results: Using the self-trained ViT encoder as the backbone and random forest, we demonstrate an accuracy of 0.88 on a weakly supervised task with all samples. The performance is significantly improved when compared to ResNet-50 trained on ImageNet. In addition, saliency maps from multi-attention heads provide excellent interpretations of morphological characteristics, including tumor stroma, cell location and necrosis. With the feature embeddings extracted by ViT, 10 morphologically distinct clusters were identified. The algorithms identified morphologically similar and dissimilar clusters among tiles that represent variation in the distribution of lymphoma cells as seen in clustering results. There was a strong correlation between clusters one and four, the mutated genes, and an increased probability of poor outcomes. When this was visualized by the pathologist, cluster 4 showed a high level of necrosis in the WSI. Finally, using the segmented WSI cortex with clustering distribution, our method achieves an AUC of 0.77 for predicting the vital status of alive/dead as an outcome. Conclusions: Our pipeline effectively leverages WSI to employ machine learning and deep learning tools for disease classification and outcome prediction in DLBCL. By computationally analyzing the entire tumor landscape, it captures tumor heterogeneity and disease risk, establishing correlations between patch-level characteristics, genomic mutations and overall outcomes. The Vision Transformer (ViT) plays a pivotal role in this process, successfully identifying specific features in histopathology tissue by leveraging self-attention mechanisms, which enables precise feature extraction and accurate analysis for DLBCL classification. Notably, the ViT model achieves high performance across various tasks without the need for external labeled data, owing to its self-supervised learning capability that enables independent learning and extraction of meaningful information. Based on this discovery cohort from this dataset, we will update the analysis using a larger, independent external cohort from institutional archives, and the results will be presented at the annual meeting.

Hydrogel-Embedded Precision-Cut Lung Slices Model Lung Cancer Premalignancy Ex Vivo.

Lung cancer is the leading global cause of cancer-related deaths. Although smoking cessation is the best prevention, 50% of lung cancer diagnoses occur in people who have quit smoking. Research into treatment options for high-risk patients is constrained to rodent models, which are time-consuming, expensive, and require large cohorts. Embedding precision-cut lung slices (PCLS) within an engineered hydrogel and exposing this tissue to vinyl carbamate, a carcinogen from cigarette smoke, creates an in vitro model of lung cancer premalignancy. Hydrogel formulations are selected to promote early lung cancer cellular phenotypes and extend PCLS viability to six weeks. Hydrogel-embedded PCLS are exposed to vinyl carbamate, which induces adenocarcinoma in mice. Analysis of proliferation, gene expression, histology, tissue stiffness, and cellular content after six weeks reveals that vinyl carbamate induces premalignant lesions with a mixed adenoma/squamous phenotype. Putative chemoprevention agents diffuse through the hydrogel and induce tissue-level changes. The design parameters selected using murine tissue are validated with hydrogel-embedded human PCLS and results show increased proliferation and premalignant lesion gene expression patterns. This tissue-engineered model of human lung cancer premalignancy is the foundation for more sophisticated ex vivo models that enable the study of carcinogenesis and chemoprevention strategies.

Classic techniques used in laser microsurgery of chronic diseases of the larynx: a literature review

The topicality of the issue of treating chronic diseases of the larynx is beyond dispute. Given the increased load on the vocal apparatus, no downward trend in disease incidence is observed. The percentage of patient visits to an otorhinolaryngologist with complaints of prolonged changes in voice timbre, vocal fatigue, and difficulties with voice control is 7–12%. Chronic productive formations of the larynx account for up to 55–70% of pathologies in the proliferative upper respiratory disease pattern. If not treated promptly, chronic diseases of the larynx can significantly affect a person’s ability to speak, swallow and breathe normally. Today, various methods for surgical treatment of chronic diseases of the larynx are used: both classical cold instruments and high-energy laser systems. The history of laser medicine dates from the late 1960s and is associated with the introduction of a high-intensity device emitting a photon stream in the infrared range into medical practice. The first carbon dioxide lasers were difficult to use (they were large in size and had complex technical settings), but at the same time they showed high resection and coagulation qualities, and it earned them their popularity. Over 3 thousand new laser devices representing a practical interest for laryngeal surgery have appeared on the medical market for more than 60-year period. Despite the fact that fundamental research demonstrates the positive results of phonosurgery with high-power devices, many otolaryngologists continue be in awe of lasers. The article presents the results of the use of traditional devices in phonosurgery for chronic diseases of the larynx and draws conclusions about the effectiveness and safety of high-power devices.

TMIC-49. AUTOMATIC SEGMENTATION OF DISTINCT GLIOBLASTOMA HALLMARKS ON DIGITIZED H&amp;E SLIDES VIA DEEP LEARNING: CURATION OF A LARGE MULTI-INSTITUTIONAL COHORT

Abstract PURPOSE Glioblastoma (GBM) is a heterogeneous brain tumor. Standard treatment constitutes surgery, chemo-radiation, and chemotherapy. After surgery, the resected tissue is analyzed under a microscope by neuropathologists to identify distinct histological hallmarks. However, visual analysis of GBM pathological features on Hematoxylin and Eosin (H&amp;E)-stained slides is subjective, time-consuming, and more importantly, requires wide expertise. We present a hierarchical deep learning-strategy to automatically segment distinct GBM hallmarks that first distinguishes necrosis (NE) from cellular tumor (CT), and subsequently, segments regions of microvascular proliferation (MV) and pseudo-palisading cell patterns (PC) within the CT region, on digitized H&amp;E slides. METHODS We employed n=541 publicly available studies including IvyGap (n=41), and TCGA (n=500). For the IvyGap cohort, expert-driven segmentations of CT, NE, MV and PC were available. We randomly employed n=21 slides from 4 patients for training, n=5 slides from 2 cases for validation, and n=2 slides from 2 cases for testing. We employed an EfficientNet model using ~600 patches of 224×224 pixels of NE and CT sampled for every slide. For semantic segmentation of MV and PC, we separately trained two EfficientNet-UNet models trained on ~200 squared-patches of 1792 and 896 pixels size, respectively. RESULTS Our hierarchical deep learning model on the IvyGap cohort achieved an accuracy of 96%, with an area under the curve (AUC) of 98% for distinguishing necrosis from non-necrotic (i.e. CT) regions. For segmenting PC and MV within the non-necrotic regions, our DL models achieve a dice score of 0.69 and 0.77, respectively. Additionally, the segmentation results for each of the pathological attributes were also generated and vetted for the TCGA cohort. CONCLUSION We developed a hierarchical model for automatic segmentation of NE, CT, and MP and PC in H&amp;E-stained Glioblastoma images which could reduce cost, time, and effort in annotating these pathological phenotypes, for quantifying the GBM micro-environment.

Impaired T-Cell Proliferation in Chronic Lymphocytic Leukemia Patients Is Linked to Disturbed Lipid Metabolism and Homeostasis

Background Successful T-cell based anti-cancer therapies require proliferation and longevity of effector T cells, features that highly depend on metabolic processes. T cells from chronic lymphocytic leukemia (CLL) patients are characterized by acquired T-cell dysfunction, resulting in impaired activation, proliferation and cytotoxicity upon in-vitro activation (van Bruggen et al, Blood 2019). Whilst the role of glucose metabolism in activated T-cells is extensively studied, the contribution of other energy sources is less clear. Lipid metabolism has only recently come into focus as an important source of energy for immune cells. Aims Molecular and functional analysis of the lipid metabolic pathway in T cells from CLL patients. Methods Peripheral blood mononuclear cells (PBMCs) of untreated CLL patients and, as controls, age-matched healthy donors (HD) were studied. T cells at baseline and after TCR stimulation were analyzed molecularly using transcriptomics and lipidomics, and extensively characterized at protein level by flow cytometry and confocal imaging. Lipid metabolism was manipulated through culturing cells in lipoprotein deficient serum (LPDS), or in the presence of agonists and inhibitors of specific regulatory pathways. Results We found the uptake of lipids, specifically cholesterol, during T-cell activation essential for proliferation, which was evidenced by a nearly absent proliferative capacity of HD T cells in the lack of exogenous lipids which was fully rescued by re-addition of cholesterol-rich low-density lipoproteins (LDL). As CLL T cells portray an altered proliferation pattern even when cultured in normal medium, transcriptomic analysis was performed to investigate the expression of key lipid metabolism genes. CLL T-cells had reduced expression of the master transcription factors SREBP1/2, governing lipid and cholesterol metabolism, when stimulated in the presence of their autologous CLL cells (Figure 1A). Accordingly, downstream protein involved in the synthesis and desaturation of lipids and LDL uptake (specifically FASN, LDL-R and SCD) were significantly reduced in CLL compared to HD T cells. Remarkably, despite reduced capacity for LDL uptake, an accumulation of neutral lipids was found in CLL T cells especially upon stimulation (Figure 1B), a feature earlier described in dysfunctional macrophages. The lipid accumulation observed in CLL T cells was supported by the lack of expression of ATGL, a crucial enzyme for the utilization of neutral lipids for energy production and building of new membranes. In line with this, mitochondrial lipid oxidation was decreased in stimulated CLL T cells compared to HD. To investigate the composition of the accumulated lipids, lipidomics analysis was performed, revealing differential abundance of specific lipid classes in stimulated CLL versus HD T cells. Triglycerides were increased while cholesterol esters and phospholipids were reduced in CLL T cells. Triglycerides likely constitute the neutral lipids accumulation found in these cells. As cholesterol esters and phospholipids are essential for proper membrane formation, reduced levels of these species can contribute to the reduced proliferative capacity of CLL T cells. Summary/conclusion Altogether, the results presented here demonstrate a disturbed lipid composition and utilization in CLL T-cells, which contributes to T-cell dysfunction on at least two levels: decreased bioenergetics and decreased proliferation. Likewise, lipid deprivation due to both decreased uptake and a misbalanced lipidome leads to altered T-cell function. Therefore, strategies to increase lipid uptake, synthesis and utilization in order to rebalance homeostasis could ameliorate CLL T-cell dysfunction and specifically proliferation-defects in CLL.

Spatial-temporal proliferation of hepatocytes during pregnancy revealed by genetic lineage tracing.

The maternal liver undergoes dramatic enlargement to adapt to the increased metabolic demands during pregnancy. However, the cellular sources for liver growth during pregnancy remain largely elusive. Here, we employed a proliferation recording system, ProTracer, to examine the spatial-temporal proliferation of hepatocytes during pregnancy. We discovered that during early to late pregnancy, hepatocyte proliferation initiated from zone 1, to zone 2, and lastly to zone 3, with the majority of new hepatocytes being generated in zone 2. Additionally, using single-cell RNA sequencing, we observed that Ccnd1 was highly enriched in zone 2 hepatocytes. We further applied dual-recombinase-mediated genetic lineage tracing to reveal that Ccnd1+ hepatocytes expanded preferentially during pregnancy. Moreover, we demonstrated that estrogen induces liver enlargement during pregnancy, which was abolished in Ccnd1 knockout mice. Our work revealed a unique spatial-temporal hepatocyte proliferation pattern during pregnancy, with Ccnd1+ hepatocytes in zone 2 serving as the major cellular source for hepatic enlargement.

Repeated mild traumatic brain injury causes sex-specific increases in cell proliferation and inflammation in juvenile rats

Childhood represents a period of significant growth and maturation for the brain, and is also associated with a heightened risk for mild traumatic brain injuries (mTBI). There is also concern that repeated-mTBI (r-mTBI) may have a long-term impact on developmental trajectories. Using an awake closed head injury (ACHI) model, that uses rapid head acceleration to induce a mTBI, we investigated the acute effects of repeated-mTBI (r-mTBI) on neurological function and cellular proliferation in juvenile male and female Long-Evans rats. We found that r-mTBI did not lead to cumulative neurological deficits with the model. R-mTBI animals exhibited an increase in BrdU + (bromodeoxyuridine positive) cells in the dentate gyrus (DG), and that this increase was more robust in male animals. This increase was not sustained, and cell proliferation returning to normal by PID3. A greater increase in BrdU + cells was observed in the dorsal DG in both male and female r-mTBI animals at PID1. Using Ki-67 expression as an endogenous marker of cellular proliferation, a robust proliferative response following r-mTBI was observed in male animals at PID1 that persisted until PID3, and was not constrained to the DG alone. Triple labeling experiments (Iba1+, GFAP+, Brdu+) revealed that a high proportion of these proliferating cells were microglia/macrophages, indicating there was a heightened inflammatory response. Overall, these findings suggest that rapid head acceleration with the ACHI model produces an mTBI, but that the acute neurological deficits do not increase in severity with repeated administration. R-mTBI transiently increases cellular proliferation in the hippocampus, particularly in male animals, and the pattern of cell proliferation suggests that this represents a neuroinflammatory response that is focused around the mid-brain rather than peripheral cortical regions. These results add to growing literature indicating sex differences in proliferative and inflammatory responses between females and males. Targeting proliferation as a therapeutic avenue may help reduce the short term impact of r-mTBI, but there may be sex-specific considerations.