Hasil untuk "Cytology"

Fernando Guerra, Cintia Giménez, Adriana Esther Rocher et al.

Background: Cervical cancer has two main features: the presence of an oncogenic virus (human papillomavirus, HPV) and the progression of infection to a low-grade squamous intraepithelial lesion (LSIL), which subsequently develops into a high-grade squamous intraepithelial lesion (HSIL) and finally into cervical cancer. In 2014, our laboratory published a study of 75 patients who were diagnosed with LSIL and underwent a second biopsy one year later. In 20 patients, the disease normalized, while in the other 55 patients, LSIL persisted or progressed to HSIL. To measure the prediction of progression, the cells were stained with AgNOR, a proliferation marker that precipitates silver at the sites of nucleolar organizer regions (NORs). In this study, a cutoff value of 3.00 μm2 was set, below which the lesions returned to normal. Materials and Methods: To confirm previous results, the aim of this work is to implement new and modern machine learning algorithms to redefine this cutoff value by applying AgNOR to LSIL. In Python, we ran two models, Support Vector Machine and Logistic Regression, to set a cutoff value for our NOR areas. Results: The cutoff value for each model was 2.82 μm2 for the Support Vector Machine and 2.78 μm2 for Logistic Regression, which are similar to the 3.00 μm2 value reported by our group in 2014. These modern algorithms represent a novel tool that can be used in women with LSIL to predict the future of this lesion. Conclusion: It is possible to predict the progression of LSIL to a persistent lesion or HSIL by staining with the AgNOR technique, measuring the NOR area, and applying machine learning algorithms.

Yeongrok Lee, Myung Jin Ban, Do Hyeon Kim et al.

<b>Background/Objectives</b>: This study aimed to evaluate the effectiveness of core needle biopsy (CNB) by comparing its diagnostic yield to fine needle aspiration cytology (FNAC) across primary and secondary examinations. <b>Methods</b>: This retrospective review analyzed medical records of patients who visited Soonchunhyang University Cheonan Hospital between January 2021 and August 2023 for thyroid nodule evaluation. Demographic data and the malignancy risk of thyroid nodules were collected based on the 2021 Korean Thyroid Imaging Reporting and Data System. FNAC and CNB results, classified using the Bethesda system for reporting thyroid cytopathology and diagnostic categories for thyroid CNB, were categorized as either “conclusive” or “inconclusive.” The rates of conclusive results in the primary examination and nodules transitioning from inconclusive to conclusive results during the secondary examination were analyzed. Finally, the diagnostic yields of FNAC and CNB were assessed using histopathological findings from surgically excised nodules. <b>Results</b>: The rate of nodules classified as “conclusive” was significantly higher in the CNB group than that in the FNAC group. Among nodules subjected to secondary examination, only the group with FNAC followed by CNB demonstrated a significant improvement in the rate of transition from inconclusive to conclusive results. Although FNAC and CNB showed comparable sensitivity and accuracy, the specificity of CNB was greater than that of FNAC. <b>Conclusions</b>: This study confirms the clinical utility of CNB by demonstrating its higher rate of conclusive results than FNAC. Future prospective studies, including cost–benefit analyses, are warranted to further define the indications for CNB.

Syunta Makabe, Kyoka Hoshi, Hiromi Kaneko et al.

Abstract Muscle-invasive bladder cancer (MIBC) is an aggressive malignancy with high recurrence and poor survival, accounting for the majority of bladder cancer–related deaths. A subset of MIBC harbors FGFR1 amplification or overexpression, associated with increased proliferation and poor prognosis. Although the pan-FGFR inhibitor erdafitinib has demonstrated clinical benefit in patients with FGFR3/FGFR2 alterations, primarily in non-MIBC, its efficacy is limited by resistance and toxicity. Moreover, its effectiveness in FGFR1-driven MIBC remains unclear. To address this gap, we investigated erdafitinib response and resistance mechanisms in JMSU1 cells, a model of FGFR1-amplified MIBC. While erdafitinib initially suppressed tumor growth, prolonged treatment led to resistance, characterized by persistent activation of ERK, AKT, and STAT1 signaling pathways. Mechanistic studies identified MET activation, driven by MET gene amplification, as a key driver of resistance. Notably, exogenous hepatocyte growth factor (HGF) not only induced resistance but also accelerated the emergence of MET-amplified, HGF-independent subpopulations under drug pressure. We also identified SHP2 as a critical mediator of FGFR1-driven ERK activation in parental cells. In resistant cells, MET activation enhanced SHP2–ERK signaling through the adaptor protein GAB1, reinforcing the resistant phenotype. Combined inhibition of FGFR1 and MET significantly suppressed tumor growth in resistant cells. These findings establish MET amplification and GAB1–SHP2 signaling as central mediators of erdafitinib resistance in FGFR1-amplified MIBC and support dual FGFR1/MET targeting as a promising therapeutic strategy.

Agata Czaplicka, Mieszko Lachota, Leszek Pączek et al.

Chimeric antigen receptor (CAR) T-cell therapy has revolutionized the treatment of malignant and non-malignant disorders. CARs are synthetic transmembrane receptors expressed on genetically modified immune effector cells, including T cells, natural killer (NK) cells, or macrophages, which are able to recognize specific surface antigens on target cells and eliminate them. CAR-modified immune cells mediate cytotoxic antitumor effects via numerous mechanisms, including the perforin and granzyme pathway, Fas and Fas Ligand (FasL) pathway, and cytokine secretion. High hopes are associated with the prospective use of the CAR-T strategy against solid cancers, especially the ones resistant to standard oncological therapies, such as pancreatic cancer (PC). Herein, we summarize the current pre-clinical and clinical studies evaluating potential tumor-associated antigens (TAA), CAR-T cell toxicities, and their efficacy in PC.

Anne-Sophie Gary, Sophie Amouret, Alicia Montoni et al.

Abstract Ultraviolet radiation (UVR) is a major environmental mutagen. In skin, UVR can initiate cancer through the induction of mutagenic DNA damage and promote its progression. An important cancer prevention mechanism is the regulated cell death (RCD), which can safely dispose of damaged cells. Apoptosis, a well-known RCD, is known to be activated by UVR, but part of the mechanism and proteins involved in UVR-induced apoptosis are still to be discovered. Receptor-interacting serine/threonine-protein kinase 3 (RIPK3) and mixed lineage kinase domain-like (MLKL) are two proteins involved in necroptosis, a form of RCD. Here, we have evaluated the implication of RIPK3 and MLKL in UVB-induced cell death in human diploid dermal fibroblasts. Our results show that RIPK3 and MLKL play opposite roles in UVB-induced cell death, in a necroptosis independent pathway. We showed that RIPK3 protects cells from UVB cell death, while MLKL sensitizes cells to UVB-induced apoptosis. Taken together these results are the first to show the implication of RIPK3 and MLKL in survival and apoptosis, respectively, bringing two new actors in UVB-induced cell death pathway.

Nicola Iacomino, Maria Cristina Tarasco, Alessia Berni et al.

Myasthenia gravis (MG) is an antibody-mediated autoimmune disorder characterized by altered neuromuscular transmission, which causes weakness and fatigability in the skeletal muscles. The etiology of MG is complex, being associated with multiple genetic and environmental factors. Over recent years, progress has been made in understanding the immunological alterations implicated in the disease, but the exact pathogenesis still needs to be elucidated. A pathogenic interplay between innate immunity and autoimmunity contributes to the intra-thymic MG development. Epigenetic changes are critically involved in both innate and adaptive immune response regulation. They can act as (i) pathological factors besides genetic predisposition and (ii) co-factors contributing to disease phenotypes or patient-specific disease course/outcomes. This article reviews the role of non-coding RNAs (ncRNAs) as epigenetic factors implicated in MG. Particular attention is dedicated to microRNAs (miRNAs), whose expression is altered in MG patients’ thymuses and circulating blood. The long ncRNA (lncRNA) contribution to MG, although not fully characterized yet, is also discussed. By summarizing the most recent and fast-growing findings on ncRNAs in MG, we highlight the therapeutic potential of these molecules for achieving immune regulation and their value as biomarkers for the development of personalized medicine approaches to improve disease care.

Elisabetta Valentini, Marta Di Martile, Matteo Brignone et al.

Abstract BH3 mimetics, targeting the Bcl-2 family anti-apoptotic proteins, represent a promising therapeutic opportunity in cancers. ABT-199, the first specific Bcl-2 inhibitor, was approved by FDA for the treatment of several hematological malignancies. We have recently discovered IS21, a novel pan BH3 mimetic with preclinical antitumor activity in several tumor types. Here, we evaluated the efficacy of IS21 and other BH3 mimetics, both as single agents and combined with the currently used antineoplastic agents in T-cell acute lymphoblastic leukemia, ovarian cancer, and melanoma. IS21 was found to be active in T-cell acute lymphoblastic leukemia, melanoma, lung, pancreatic, and ovarian cancer cell lines. Bcl-xL and Mcl-1 protein levels predicted IS21 sensitivity in melanoma and ovarian cancer, respectively. Exploring IS21 mechanism of action, we found that IS21 activity depends on the presence of BAX and BAK proteins: complexes between Bcl-2 and Bcl-xL proteins and their main binding partners were reduced after IS21 treatment. In combination experiments, BH3 mimetics sensitized leukemia cells to chemotherapy, ovarian cancer cells and melanoma models to PARP and MAPK inhibitors, respectively. We showed that this enhancing effect was related to the potentiation of the apoptotic pathway, both in hematologic and solid tumors. In conclusion, our data suggest the use of inhibitors of anti-apoptotic proteins as a therapeutic strategy to enhance the efficacy of anticancer treatment.

Gulnara Svishcheva, Olga Babayan, Taras Sipko et al.

Rangifer tarandus L. 1758 is one of the few modern hoofed species in which domestic and wild forms coexist in the same territory. The genetic differentiation of domestic and wild reindeer in Northern Eurasia was examined using microsatellite data. А total of 780 animals were studied at 16 microsatellite loci. Samples of wild reindeer were taken from seven populations inhabiting different natural areas, and samples of domestic animals were selected from the Evenki, Evens, Chukchi and Nenets breeds, including two ecotypes, Tofalar and Todzha reindeer. The levels of genetic diversity and variation in wild reindeer were higher than in domestic ones. Bayesian clustering analysis allowed us to distinguish domesticated reindeer populations by the degree of taming, but failed to detect differences in genetic structure between wild reindeer populations. These differences were found using the pairwise Fst values. Overall, the microsatellite analysis revealed a significant genetic differentiation between domestic and wild forms and the structuring of populations within each form, which may be important for the development of strategies for animal conservation.

Nadine Hosny El Said, Francesco Della Valle, Peng Liu et al.

Abstract PRC2-mediated epigenetic function involves the interaction with long non-coding RNAs (lncRNAs). Although the identity of some of these RNAs has been elucidated in the context of developmental programs, their counterparts in postmitotic adult tissue homeostasis remain uncharacterized. To this aim, we used terminally differentiated postmitotic skeletal muscle cells in which oxidative stress induces the dynamic activation of PRC2-Ezh1 through Embryonic Ectoderm Develpment (EED) shuttling to the nucleus. We identify lncRNA Malat-1 as a necessary partner for PRC2-Ezh1-dependent response to oxidative stress. We show that in this pathway, PRC2-EZH1 dynamic assembly, and in turn stress induced skeletal muscle targeted genes repression, depends specifically on Malat-1. Our study reports about PRC2–RNA interactions in the physiological context of adaptive oxidative stress response and identifies the first lncRNA involved in PRC2-Ezh1 function.

Alessandro Sacco, Anna Martina Battaglia, Cirino Botta et al.

New insights into the field of iron metabolism within the tumor microenvironment have been uncovered in recent years. Iron promotes the production of reactive oxygen species, which may either trigger ferroptosis cell death or contribute to malignant transformation. Once transformed, cancer cells divert tumor-infiltrating immune cells to satisfy their iron demand, thus affecting the tumor immunosurveillance. In this review, we highlight how the bioavailability of this metal shapes complex metabolic pathways within the tumor microenvironment and how this affects both tumor-associated macrophages and tumor-infiltrating lymphocytes functions. Furthermore, we discuss the potentials as well as the current clinical controversies surrounding the use of iron metabolism as a target for new anticancer treatments in two opposed conditions: (i) the “hot” tumors, which are usually enriched in immune cells infiltration and are extremely rich in iron availability within the microenvironment, and (ii) the “cold” tumors, which are often very poor in immune cells, mainly due to immune exclusion.

María José Rodríguez, Matías Sabaj, Gerardo Tolosa et al.

Liver fibrosis is a complex process characterized by the excessive accumulation of extracellular matrix (ECM) and an alteration in liver architecture, as a result of most types of chronic liver diseases such as cirrhosis, hepatocellular carcinoma (HCC) and liver failure. Maresin-1 (MaR1) is derivative of ω-3 docosahexaenoic acid (DHA), which has been shown to have pro-resolutive and anti-inflammatory effects. We tested the hypothesis that the application of MaR1 could prevent the development of fibrosis in an animal model of chronic hepatic damage. Sprague-Dawley rats were induced with liver fibrosis by injections of diethylnitrosamine (DEN) and treated with or without MaR1 for four weeks. In the MaR1-treated animals, levels of AST and ALT were normalized in comparison with DEN alone, the hepatic architecture was improved, and inflammation and necrotic areas were reduced. Cell proliferation, assessed by the mitotic activity index and the expression of Ki-67, was increased in the MaR1-treated group. MaR1 attenuated liver fibrosis and oxidative stress was induced by DEN. Plasma levels of the pro-inflammatory mediators TNF-α and IL-1β were reduced in MaR1-treated animals, whereas the levels of IL-10, an anti-inflammatory cytokine, increased. Interestingly, MaR1 inhibited the translocation of the p65 subunit of NF-κB, while increasing the activation of Nrf2, a key regulator of the antioxidant response. Finally, MaR1 treatment reduced the levels of the pro-fibrotic mediator TGF-β and its receptor, while normalizing the hepatic levels of IGF-1, a proliferative agent. Taken together, these results suggest that MaR1 improves the parameters of DEN-induced liver fibrosis, activating hepatocyte proliferation and decreasing oxidative stress and inflammation. These results open the possibility of MaR1 as a potential therapeutic agent in fibrosis and other liver pathologies.

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Olga Akopova, Liudmila Kolchinskaya, Valentina Nosar et al.

Abstract Background Cytoprotection afforded by mitochondrial ATP-sensitive K+-channel (mKATP-channel) opener diazoxide (DZ) largely depends on the activation of potassium cycle with eventual modulation of mitochondrial functions and ROS production. However, generally these effects were studied in the presence of Mg∙ATP known to block K+ transport. Thus, the purpose of our work was the estimation of DZ effects on K+ transport, K+ cycle and ROS production in rat liver mitochondria in the absence of Mg∙ATP. Results Without Mg·ATP, full activation of native mKATP-channel, accompanied by the increase in ATP-insensitive K+ uptake, activation of K+-cycle and respiratory uncoupling, was reached at ≤0.5 μM of DZ,. Higher diazoxide concentrations augmented ATP-insensitive K+ uptake, but not mKATP-channel activity. mKATP-channel was blocked by Mg·ATP, reactivated by DZ, and repeatedly blocked by mKATP-channel blockers glibenclamide and 5-hydroxydecanoate, whereas ATP-insensitive potassium transport was blocked by Mg2+ and was not restored by DZ. High sensitivity of potassium transport to DZ in native mitochondria resulted in suppression of mitochondrial ROS production caused by the activation of K+-cycle on sub-micromolar scale. Based on the oxygen consumption study, the share of mKATP-channel in respiratory uncoupling by DZ was found. Conclusions The study of mKATP-channel activation by diazoxide in the absence of MgATP discloses novel, not described earlier, aspects of mKATP-channel interaction with this drug. High sensitivity of mKATP-channel to DZ results in the modulation of mitochondrial functions and ROS production by DZ on sub-micromolar concentration scale. Our experiments led us to the hypothesis that under the conditions marked by ATP deficiency affinity of mKATP-channel to DZ can increase, which might contribute to the high effectiveness of this drug in cardio- and neuroprotection.

Diego Fresegna, Silvia Bullitta, Alessandra Musella et al.

Multiple sclerosis (MS) is a common neurological disorder of putative autoimmune origin. Clinical and experimental studies delineate abnormal expression of specific cytokines over the course of the disease. One major cytokine that has been shown to play a pivotal role in MS is tumor necrosis factor (TNF). TNF is a pleiotropic cytokine regulating many physiological and pathological functions of both the immune system and the central nervous system (CNS). Convincing evidence from studies in human and experimental MS have demonstrated the involvement of TNF in various pathological hallmarks of MS, including immune dysregulation, demyelination, synaptopathy and neuroinflammation. However, due to the complexity of TNF signaling, which includes two-ligands (soluble and transmembrane TNF) and two receptors, namely TNF receptor type-1 (TNFR1) and type-2 (TNFR2), and due to its cell- and context-differential expression, targeting the TNF system in MS is an ongoing challenge. This review summarizes the evidence on the pathophysiological role of TNF in MS and in different MS animal models, with a special focus on pharmacological treatment aimed at controlling the dysregulated TNF signaling in this neurological disorder.

Matteo Petini, Tommaso Furlanello, Patrizia Danesi et al.

A 7-month-old Cavalier King Charles Spaniel female was referred due to a chronic cough refractory to antibiotic treatments. Laboratory findings showed leukocytosis, increased serum C-reactive protein, hypogammaglobulinemia, and decreased total serum immunoglobulin G concentration. Thoracic radiographs showed a mild bronchial pattern. Cytology of the bronchoalveolar lavage fluid revealed a septic inflammation. Bordetella bronchiseptica, Mycoplasma spp., and Pneumocystis carinii were identified by polymerase chain reaction testing, and Klebsiella pneumonia was cultured from the bronchoalveolar lavage fluid. Moreover, Escherichia coli was also cultured from urine. Pneumocystis spp. identification was done by sequencing of genetic amplicons. The dog died due to cardiopulmonary arrest secondary to a spontaneous pneumothorax on the day following the procedure. This report documents the detection of Pneumocystis carinii f. sp. canis in a suspected immunocompromised Cavalier King Charles Spaniel with concurrent pulmonary and urinary tract infections involving four different pathogens, and highlights the importance of the use of polymerase chain reaction testing to detect canine Pneumocystis spp. in cases with negative bronchoalveolar lavage cytology.

Rongzu Wu, Tingchun Wu, Kai Wang et al.

Abstract Background The prognostic significance of galectin-1 (Gal-1) expression in cancerous patients has been assessed for several years while the results remain controversial. Thus, we performed the first comprehensive meta-analysis to evaluate the prognostic value of Gal-1 expression in cancerous patients. Methods We searched Pubmed, Embase and Web of Science to recruit studies on the prognostic impact of Gal-1 expression in cancerous patients. Eighteen studies containing 2674 patients were involved in this meta-analysis until March 30, 2018. Pooled hazard ratios (HRs) with 95% confidence interval (95% CI) were calculated to estimate the effect using random-effects model. Results The pooled results revealed that high Gal-1 expression in cancer tissue associated with a poor OS (HR = 1.79, 95% CI 1.54–2.08, P < 0.001). In the subgroup of tumor type, it’s observed that high Gal-1 expression was significant correlated with poor OS in digestive cancers without heterogeneity (HR = 1.94, 95% CI 1.64–2.30, P < 0.001; fixed-effects model; I2 = 20.1%, P = 0.276). Conclusions Our present meta-analysis indicates that high Gal-1 expression might be a predictive factor of poor prognosis in cancers, particularly in digestive cancers.

Sharon Y Y Low, Chen Min Wei, Kenneth T E Chang et al.

Leptomeningeal disease is a feared sequelae of malignant paediatric brain tumours. Current methods for its detection is the combined use of cranio-spinal MRI, and CSF cytology from a post-operative lumbar puncture. In this study, the authors hypothesize that CSF taken at the start of surgery, either from an external ventricular drain or neuroendoscope will have equal sensitivity for positive tumour cells, in comparison to lumbar puncture. Secondary hypotheses include positive correlation between CSF cytology and MRI findings of LMD. From a clinical perspective, the key aim of the study was for affected paediatric patients to avoid an additional procedure of a lumbar puncture, often performed under anaesthesia after neurosurgical intervention.This is single-institution, retrospective study of paediatric patients diagnosed with malignant brain tumours. Its main aim was to compare cytological data from CSF collected at the time of surgery versus data from an interval lumbar puncture. In addition, MRI imaging of the same cohort of patients was examined for leptomeningeal disease and corroborated against CSF tumour cytology findings.Thirty patients are recruited for this study. Data analysis demonstrates a statistically significant association between our intra-operative CSF and LP sampling. Furthermore, our results also show for significant correlation between evidence of leptomeningeal disease on MRI findings versus intra-operative CSF positivity for tumour cells.Although this is a retrospective study with a limited population, our data concurs with potential to avoid an additional procedure for the paediatric patient diagnosed with a malignant brain tumour.

Rana S. Hoda, Christopher VandenBussche, Syed A. Hoda

N. Linta, M. Baron Toaldo, G. Bettini et al.

Abstract Background This study describes the feasibility of Contrast Enhanced Ultrasonography (CEUS) in the diagnostic work-up of non-cardiac thoracic disorders of small animals. The second aim is to assess the usefulness of CEUS as a direct guide for sample procedures. Results Forty animals, 28 dogs and 12 cats, were included in the study. Thoracic disorders included 23 pulmonary lesions [primary carcinoma (14), lymphoma (1), sarcoma (1), histiocytic sarcoma (1), abscess (1) and pneumonia (5)] and 17 mediastinal lesions [lymphoma (8), thymoma (3), mesothelioma (1), melanoma (1), carcinomatous lymphadenopathy (1), mixsosarcoma (1), lipoma (1), and abscess (1)]. The majority of neoplastic pulmonary lesions showed an inhomogeneous distribution of contrast medium, whereas inflammatory lesions had a homogenous distribution with typical pulmonary vessels ramification. The majority of mediastinal malignant lesions showed an inhomogeneous distribution pattern. The lung and mediastinal abscesses had peripheral enhancement of the wall with an avascular center. All cytological and biopsy samples obtained after CEUS were diagnostic. Quantitative analysis, performed in 19/23 pulmonary lesions, showed a statistically significant difference (P < 0.0001) between the arrival time of the malignant (7.27 s - range 4.46–13.52 s) and benign (4.52 s - range 2.87–6.06 s) pulmonary lesions. Conclusions CEUS may be a useful tool for the evaluation of non-cardiac thoracic lesions. The contrast medium allows for the precise definition of lesion edges, the presence of necrotic areas, and the distribution of pulmonary vessels. Based on our preliminary results, the use of ultrasonographic contrast medium can be recommended for improving the diagnostic usefulness of cytology and biopsy sampling, because CEUS may help to define necrotic areas from viable tissue.