69 results on '"Francesco Paolo Cammarata"'
Search Results
2. Biodistribution Assessment of a Novel 68Ga-Labeled Radiopharmaceutical in a Cancer Overexpressing CCK2R Mouse Model: Conventional and Radiomics Methods for Analysis
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Anna Maria Pavone, Viviana Benfante, Paolo Giaccone, Alessandro Stefano, Filippo Torrisi, Vincenzo Russo, Davide Serafini, Selene Richiusa, Marco Pometti, Fabrizio Scopelliti, Massimo Ippolito, Antonino Giulio Giannone, Daniela Cabibi, Mattia Asti, Elisa Vettorato, Luca Morselli, Mario Merone, Marcello Lunardon, Alberto Andrighetto, Antonino Tuttolomondo, Francesco Paolo Cammarata, Marco Verona, Giovanni Marzaro, Francesca Mastrotto, Rosalba Parenti, Giorgio Russo, and Albert Comelli
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68Ga-labeled radiopharmaceutical ,biodistribution ,micro-PET/CT ,mouse imaging ,radiomics ,nuclear medicine ,Science - Abstract
The aim of the present study consists of the evaluation of the biodistribution of a novel 68Ga-labeled radiopharmaceutical, [68Ga]Ga-NODAGA-Z360, injected into Balb/c nude mice through histopathological analysis on bioptic samples and radiomics analysis of positron emission tomography/computed tomography (PET/CT) images. The 68Ga-labeled radiopharmaceutical was designed to specifically bind to the cholecystokinin receptor (CCK2R). This receptor, naturally present in healthy tissues such as the stomach, is a biomarker for numerous tumors when overexpressed. In this experiment, Balb/c nude mice were xenografted with a human epidermoid carcinoma A431 cell line (A431 WT) and overexpressing CCK2R (A431 CCK2R+), while controls received a wild-type cell line. PET images were processed, segmented after atlas-based co-registration and, consequently, 112 radiomics features were extracted for each investigated organ / tissue. To confirm the histopathology at the tissue level and correlate it with the degree of PET uptake, the studies were supported by digital pathology. As a result of the analyses, the differences in radiomics features in different body districts confirmed the correct targeting of the radiopharmaceutical. In preclinical imaging, the methodology confirms the importance of a decision-support system based on artificial intelligence algorithms for the assessment of radiopharmaceutical biodistribution.
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- 2024
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3. An Overview of In Vitro Assays of 64Cu-, 68Ga-, 125I-, and 99mTc-Labelled Radiopharmaceuticals Using Radiometric Counters in the Era of Radiotheranostics
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Viviana Benfante, Alessandro Stefano, Muhammad Ali, Riccardo Laudicella, Walter Arancio, Antonino Cucchiara, Fabio Caruso, Francesco Paolo Cammarata, Claudia Coronnello, Giorgio Russo, Monica Miele, Alessandra Vieni, Antonino Tuttolomondo, Anthony Yezzi, and Albert Comelli
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in vitro test ,radiopharmaceuticals radiobiology ,gamma counter ,radiotheranostics ,imaging ,cancer ,Medicine (General) ,R5-920 - Abstract
Radionuclides are unstable isotopes that mainly emit alpha (α), beta (β) or gamma (γ) radiation through radiation decay. Therefore, they are used in the biomedical field to label biomolecules or drugs for diagnostic imaging applications, such as positron emission tomography (PET) and/or single-photon emission computed tomography (SPECT). A growing field of research is the development of new radiopharmaceuticals for use in cancer treatments. Preclinical studies are the gold standard for translational research. Specifically, in vitro radiopharmaceutical studies are based on the use of radiopharmaceuticals directly on cells. To date, radiometric β- and γ-counters are the only tools able to assess a preclinical in vitro assay with the aim of estimating uptake, retention, and release parameters, including time- and dose-dependent cytotoxicity and kinetic parameters. This review has been designed for researchers, such as biologists and biotechnologists, who would like to approach the radiobiology field and conduct in vitro assays for cellular radioactivity evaluations using radiometric counters. To demonstrate the importance of in vitro radiopharmaceutical assays using radiometric counters with a view to radiogenomics, many studies based on 64Cu-, 68Ga-, 125I-, and 99mTc-labeled radiopharmaceuticals have been revised and summarized in this manuscript.
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- 2023
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4. The Proton-Boron Reaction Increases the Radiobiological Effectiveness of Clinical Low- and High-Energy Proton Beams: Novel Experimental Evidence and Perspectives
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Pavel Bláha, Chiara Feoli, Stefano Agosteo, Marco Calvaruso, Francesco Paolo Cammarata, Roberto Catalano, Mario Ciocca, Giuseppe Antonio Pablo Cirrone, Valeria Conte, Giacomo Cuttone, Angelica Facoetti, Giusi Irma Forte, Lorenzo Giuffrida, Giuseppe Magro, Daniele Margarone, Luigi Minafra, Giada Petringa, Gaia Pucci, Valerio Ricciardi, Enrico Rosa, Giorgio Russo, and Lorenzo Manti
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protontherapy ,chromosome aberrations ,proton-boron (B) fusion-enhanced proton therapy (PBFEPT) ,BSH ,alpha-particle ,cancer cell killing ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Protontherapy is a rapidly expanding radiotherapy modality where accelerated proton beams are used to precisely deliver the dose to the tumor target but is generally considered ineffective against radioresistant tumors. Proton-Boron Capture Therapy (PBCT) is a novel approach aimed at enhancing proton biological effectiveness. PBCT exploits a nuclear fusion reaction between low-energy protons and 11B atoms, i.e. p+11B→ 3α (p-B), which is supposed to produce highly-DNA damaging α-particles exclusively across the tumor-conformed Spread-Out Bragg Peak (SOBP), without harming healthy tissues in the beam entrance channel. To confirm previous work on PBCT, here we report new in-vitro data obtained at the 62-MeV ocular melanoma-dedicated proton beamline of the INFN-Laboratori Nazionali del Sud (LNS), Catania, Italy. For the first time, we also tested PBCT at the 250-MeV proton beamline used for deep-seated cancers at the Centro Nazionale di Adroterapia Oncologica (CNAO), Pavia, Italy. We used Sodium Mercaptododecaborate (BSH) as 11B carrier, DU145 prostate cancer cells to assess cell killing and non-cancer epithelial breast MCF-10A cells for quantifying chromosome aberrations (CAs) by FISH painting and DNA repair pathway protein expression by western blotting. Cells were exposed at various depths along the two clinical SOBPs. Compared to exposure in the absence of boron, proton irradiation in the presence of BSH significantly reduced DU145 clonogenic survival and increased both frequency and complexity of CAs in MCF-10A cells at the mid- and distal SOBP positions, but not at the beam entrance. BSH-mediated enhancement of DNA damage response was also found at mid-SOBP. These results corroborate PBCT as a strategy to render protontherapy amenable towards radiotherapy-resilient tumor. If coupled with emerging proton FLASH radiotherapy modalities, PBCT could thus widen the protontherapy therapeutic index.
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- 2021
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5. Chelation of Theranostic Copper Radioisotopes with S-Rich Macrocycles: From Radiolabelling of Copper-64 to In Vivo Investigation
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Marianna Tosato, Marco Verona, Chiara Favaretto, Marco Pometti, Giordano Zanoni, Fabrizio Scopelliti, Francesco Paolo Cammarata, Luca Morselli, Zeynep Talip, Nicholas P. van der Meulen, Valerio Di Marco, and Mattia Asti
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copper-64 ,chelators ,radiopharmaceuticals ,cancer ,Organic chemistry ,QD241-441 - Abstract
Copper radioisotopes are generally employed for cancer imaging and therapy when firmly coordinated via a chelating agent coupled to a tumor-seeking vector. However, the biologically triggered Cu2+-Cu+ redox switching may constrain the in vivo integrity of the resulting complex, leading to demetallation processes. This unsought pathway is expected to be hindered by chelators bearing N, O, and S donors which appropriately complements the borderline-hard and soft nature of Cu2+ and Cu+. In this work, the labelling performances of a series of S-rich polyazamacrocyclic chelators with [64Cu]Cu2+ and the stability of the [64Cu]Cu-complexes thereof were evaluated. Among the chelators considered, the best results were obtained with 1,7-bis [2-(methylsulfanyl)ethyl]-4,10,diacetic acid-1,4,7,10-tetraazacyclododecane (DO2A2S). DO2A2S was labelled at high molar activities in mild reaction conditions, and its [64Cu]Cu2+ complex showed excellent integrity in human serum over 24 h. Biodistribution studies in BALB/c nude mice performed with [64Cu][Cu(DO2A2S)] revealed a behavior similar to other [64Cu]Cu-labelled cyclen derivatives characterized by high liver and kidney uptake, which could either be ascribed to transchelation phenomena or metabolic processing of the intact complex.
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- 2022
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6. Evaluation of Proton-Induced Biomolecular Changes in MCF-10A Breast Cells by Means of FT-IR Microspectroscopy
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Valerio Ricciardi, Marianna Portaccio, Maria Lasalvia, Francesco Paolo Cammarata, Pietro Pisciotta, Giuseppe Perna, Vito Capozzi, Giada Petringa, Lorenzo Manti, and Maria Lepore
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MCF-10A breast cells ,Fourier transform infrared microspectroscopy ,proton therapy ,radiation-induced effects ,Technology ,Engineering (General). Civil engineering (General) ,TA1-2040 ,Biology (General) ,QH301-705.5 ,Physics ,QC1-999 ,Chemistry ,QD1-999 - Abstract
Radiotherapy (RT) with accelerated beams of charged particles (protons and carbon ions), also known as hadrontherapy, is a treatment modality that is increasingly being adopted thanks to the several benefits that it grants compared to conventional radiotherapy (CRT) treatments performed by means of high-energy photons/electrons. Hence, information about the biomolecular effects in exposed cells caused by such particles is needed to better realize the underlying radiobiological mechanisms and to improve this therapeutic strategy. To this end, Fourier transform infrared microspectroscopy (μ-FT-IR) can be usefully employed, in addition to long-established radiobiological techniques, since it is currently considered a helpful tool for examining radiation-induced cellular changes. In the present study, MCF-10A breast cells were chosen to evaluate the effects of proton exposure using μ-FT-IR. They were exposed to different proton doses and fixed at various times after exposure to evaluate direct effects due to proton exposure and the kinetics of DNA damage repair. Irradiated and control cells were examined in transflection mode using low-e substrates that have been recently demonstrated to offer a fast and direct way to examine proton-exposed cells. The acquired spectra were analyzed using a deconvolution procedure and a ratiometric approach, both of which showed the different contributions of DNA, protein, lipid, and carbohydrate cell components. These changes were particularly significant for cells fixed 48 and 72 h after exposure. Lipid changes were related to variations in membrane fluidity, and evidence of DNA damage was highlighted. The analysis of the Amide III band also indicated changes that could be related to different enzyme contributions in DNA repair.
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- 2022
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7. A New Preclinical Decision Support System Based on PET Radiomics: A Preliminary Study on the Evaluation of an Innovative 64Cu-Labeled Chelator in Mouse Models
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Viviana Benfante, Alessandro Stefano, Albert Comelli, Paolo Giaccone, Francesco Paolo Cammarata, Selene Richiusa, Fabrizio Scopelliti, Marco Pometti, Milene Ficarra, Sebastiano Cosentino, Marcello Lunardon, Francesca Mastrotto, Alberto Andrighetto, Antonino Tuttolomondo, Rosalba Parenti, Massimo Ippolito, and Giorgio Russo
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radiomics ,micro-PET/CT ,mouse imaging ,atlas ,64Cu-labeled chelator ,Photography ,TR1-1050 ,Computer applications to medicine. Medical informatics ,R858-859.7 ,Electronic computers. Computer science ,QA75.5-76.95 - Abstract
The 64Cu-labeled chelator was analyzed in vivo by positron emission tomography (PET) imaging to evaluate its biodistribution in a murine model at different acquisition times. For this purpose, nine 6-week-old female Balb/C nude strain mice underwent micro-PET imaging at three different time points after 64Cu-labeled chelator injection. Specifically, the mice were divided into group 1 (acquisition 1 h after [64Cu] chelator administration, n = 3 mice), group 2 (acquisition 4 h after [64Cu]chelator administration, n = 3 mice), and group 3 (acquisition 24 h after [64Cu] chelator administration, n = 3 mice). Successively, all PET studies were segmented by means of registration with a standard template space (3D whole-body Digimouse atlas), and 108 radiomics features were extracted from seven organs (namely, heart, bladder, stomach, liver, spleen, kidney, and lung) to investigate possible changes over time in [64Cu]chelator biodistribution. The one-way analysis of variance and post hoc Tukey Honestly Significant Difference test revealed that, while heart, stomach, spleen, kidney, and lung districts showed a very low percentage of radiomics features with significant variations (p-value < 0.05) among the three groups of mice, a large number of features (greater than 60% and 50%, respectively) that varied significantly between groups were observed in bladder and liver, indicating a different in vivo uptake of the 64Cu-labeled chelator over time. The proposed methodology may improve the method of calculating the [64Cu]chelator biodistribution and open the way towards a decision support system in the field of new radiopharmaceuticals used in preclinical imaging trials.
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- 2022
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8. Evaluation of proton beam radiation-induced skin injury in a murine model using a clinical SOBP.
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Pietro Pisciotta, Angelita Costantino, Francesco Paolo Cammarata, Filippo Torrisi, Giovanna Calabrese, Valentina Marchese, Giuseppe Antonio Pablo Cirrone, Giada Petringa, Giusi Irma Forte, Luigi Minafra, Valentina Bravatà, Massimo Gulisano, Fabrizio Scopelliti, Francesco Tommasino, Emanuele Scifoni, Giacomo Cuttone, Massimo Ippolito, Rosalba Parenti, and Giorgio Russo
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Medicine ,Science - Abstract
The purpose of this paper is to characterize the skin deterministic damage due to the effect of proton beam irradiation in mice occurred during a long-term observational experiment. This study was initially defined to evaluate the insurgence of myelopathy irradiating spinal cords with the distal part of a Spread-out Bragg peak (SOBP). To the best of our knowledge, no study has been conducted highlighting high grades of skin injury at the dose used in this paper. Nevertheless these effects occurred. In this regard, the experimental evidence of significant insurgence of skin injury induced by protons using a SOBP configuration will be shown. Skin damages were classified into six scores (from 0 to 5) according to the severity of the injuries and correlated to ED50 (i.e. the radiation dose at which 50% of animals show a specific score) at 40 days post-irradiation (d.p.i.). The effects of radiation on the overall animal wellbeing have been also monitored and the severity of radiation-induced skin injuries was observed and quantified up to 40 d.p.i.
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- 2020
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9. Radiobiological Outcomes, Microdosimetric Evaluations and Monte Carlo Predictions in Eye Proton Therapy
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Giada Petringa, Marco Calvaruso, Valeria Conte, Pavel Bláha, Valentina Bravatà, Francesco Paolo Cammarata, Giacomo Cuttone, Giusi Irma Forte, Otilija Keta, Lorenzo Manti, Luigi Minafra, Vladana Petković, Ivan Petrović, Selene Richiusa, Aleksandra Ristić Fira, Giorgio Russo, and Giuseppe Antonio Pablo Cirrone
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protontherapy ,proton ,RBE ,radiobiology ,microdosimetry ,Geant4 ,Technology ,Engineering (General). Civil engineering (General) ,TA1-2040 ,Biology (General) ,QH301-705.5 ,Physics ,QC1-999 ,Chemistry ,QD1-999 - Abstract
CATANA (Centro di AdroTerapia ed Applicazioni Nucleari Avanzate) was the first Italian protontherapy facility dedicated to the treatment of ocular neoplastic pathologies. It is in operation at the LNS Laboratories of the Italian Institute for Nuclear Physics (INFN-LNS) and to date, 500 patients have been successfully treated. Even though proton therapy has demonstrated success in clinical settings, there is still a need for more accurate models because they are crucial for the estimation of clinically relevant RBE values. Since RBE can vary depending on several physical and biological parameters, there is a clear need for more experimental data to generate predictions. Establishing a database of cell survival experiments is therefore useful to accurately predict the effects of irradiations on both cancerous and normal tissue. The main aim of this work was to compare RBE values obtained from in-vitro experimental data with predictions made by the LEM II (Local Effect Model), Monte Carlo approaches, and semi-empirical models based on LET experimental measurements. For this purpose, the 92.1 uveal melanoma and ARPE-19 cells derived from normal retinal pigmented epithelium were selected and irradiated in the middle of clinical SOBP of the CATANA proton therapy facility. The remarkable results show the potentiality of using microdosimetric spectrum, Monte Carlo simulations and LEM model to predict not only the RBE but also the survival curves.
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- 2021
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10. Sensitization of chondrosarcoma cells with PARP inhibitor and high-LET radiation
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Mathieu Césaire, Utpal Ghosh, Jean-Baptiste Austry, Etienne Muller, Francesco Paolo Cammarata, Marilyne Guillamin, Massimo Caruso, Laurent Castéra, Giada Petringa, Giuseppe Antonio Pablo Cirrone, and François Chevalier
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Diseases of the musculoskeletal system ,RC925-935 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Chondrosarcoma is a malignant tumor that arises from cartilaginous tissue and is radioresistant and chemoresistant to conventional treatments. The preferred treatment consists of surgical resection, which might cause severe disabilities for the patient; in addition, this procedure might be impossible for inoperable locations, such as the skull base. Carbon ion irradiation (hadron therapy) has been proposed as an alternative treatment, primarily due to its greater biological effectiveness and improved ballistic properties compared with conventional radiotherapy with X-rays. The goal of this study was to characterize the genetic mutations of a grade III chondrosarcoma cell line (CH2879) and examine the cellular responses to conventional radiotherapy (X-rays) and hadron therapy (proton and carbon ions) in the presence of the PARP inhibitor Olaparib. To better understand PARP inhibition, we first analyzed the formation of poly-ADP ribose chains by western blot; we observed an increase in its signal after irradiation, which disappeared on addition of the PARP inhibitor. PARPi enhanced ratio of approximately 1.3, 1.8, and 1.5 following irradiation of cells with X-rays, protons, and C-ions, respectively, as detected by clonogenic assay. The decrease in cell survival was confirmed by proliferation assay. The radiosensitivity of CH2879 cells was associated with mutations in homologous recombination repair genes, such as RAD50, SMARCA2 and NBN. This study demonstrates the capacity of the PARP inhibitor Olaparib to radiosensitize mutated chondrosarcoma cells to conventional photon irradiation, proton and carbon ion irradiation. Keywords: PARP inhibitor, Olaparib, Chondrosarcoma, Carbon ions irradiation, Mutation status
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- 2019
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11. FT-IR Transflection Micro-Spectroscopy Study on Normal Human Breast Cells after Exposure to a Proton Beam
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Valerio Ricciardi, Marianna Portaccio, Giuseppe Perna, Maria Lasalvia, Vito Capozzi, Francesco Paolo Cammarata, Pietro Pisciotta, Giada Petringa, Ines Delfino, Lorenzo Manti, and Maria Lepore
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fourier transform infrared micro-spectroscopy ,transflection geometry ,proton therapy ,radiation dose effects ,human breast cells ,Technology ,Engineering (General). Civil engineering (General) ,TA1-2040 ,Biology (General) ,QH301-705.5 ,Physics ,QC1-999 ,Chemistry ,QD1-999 - Abstract
Fourier transform infrared micro-spectroscopy (μ-FT-IR) is nowadays considered a valuable tool for investigating the changes occurring in human cells after exposure to ionizing radiation. Recently, considerable attention has been devoted to the use of this optical technique in the study of cells exposed to proton beams, that are being increasingly adopted in cancer therapy. Different experimental configurations are used for proton irradiation and subsequent spectra acquisition. To facilitate the use of μ-FT-IR, it may be useful to investigate new experimental approaches capable of speeding up and simplifying the irradiation and measurements phases. Here, we propose the use of low-e-substrates slides for cell culture, allowing the irradiation and spectra acquisition in transflection mode in a fast and direct way. In recent years, there has been a wide debate about the validity of these supports, but many researchers agree that the artifacts due to the presence of the electromagnetic standing wave effects are negligible in many practical cases. We investigated human normal breast cells (MCF-10 cell line) fixed immediately after the irradiation with graded proton radiation doses (0, 0.5, 2, and 4 Gy). The spectra obtained in transflection geometry showed characteristics very similar to those present in the spectra acquired in transmission geometry and confirm the validity of the chosen approach. The analysis of spectra indicates the occurrence of significant changes in DNA and lipids components of cells. Modifications in protein secondary structure are also evidenced.
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- 2021
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12. MF2C3: Multi-Feature Fuzzy Clustering to Enhance Cell Colony Detection in Automated Clonogenic Assay Evaluation
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Carmelo Militello, Leonardo Rundo, Luigi Minafra, Francesco Paolo Cammarata, Marco Calvaruso, Vincenzo Conti, and Giorgio Russo
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clonogenic assay ,automatic cell colony detection ,spatial fuzzy c-means clustering ,multi-feature clustering ,entropy ,standard deviation ,Mathematics ,QA1-939 - Abstract
A clonogenic assay is a biological technique for calculating the Surviving Fraction (SF) that quantifies the anti-proliferative effect of treatments on cell cultures: this evaluation is often performed via manual counting of cell colony-forming units. Unfortunately, this procedure is error-prone and strongly affected by operator dependence. Besides, conventional assessment does not deal with the colony size, which is generally correlated with the delivered radiation dose or administered cytotoxic agent. Relying upon the direct proportional relationship between the Area Covered by Colony (ACC) and the colony count and size, along with the growth rate, we propose MF2C3, a novel computational method leveraging spatial Fuzzy C-Means clustering on multiple local features (i.e., entropy and standard deviation extracted from the input color images acquired by a general-purpose flat-bed scanner) for ACC-based SF quantification, by considering only the covering percentage. To evaluate the accuracy of the proposed fully automatic approach, we compared the SFs obtained by MF2C3 against the conventional counting procedure on four different cell lines. The achieved results revealed a high correlation with the ground-truth measurements based on colony counting, by outperforming our previously validated method using local thresholding on L*u*v* color well images. In conclusion, the proposed multi-feature approach, which inherently leverages the concept of symmetry in the pixel local distributions, might be reliably used in biological studies.
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- 2020
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13. Betula etnensis Raf. (Betulaceae) Extract Induced HO-1 Expression and Ferroptosis Cell Death in Human Colon Cancer Cells
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Giuseppe Antonio Malfa, Barbara Tomasello, Rosaria Acquaviva, Carlo Genovese, Alfonsina La Mantia, Francesco Paolo Cammarata, Monica Ragusa, Marcella Renis, and Claudia Di Giacomo
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Colon cancer ,Betula etnensis Raf. ,oxidative stress ,heme oxigenase-1 ,ferroptosis ,thiol groups ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Betula etnensis Raf. (Birch Etna) belonging to the Betulaceae family grows on the eastern slope of Etna. Many bioactive compounds present in Betula species are considered promising anticancer agents. In this study, we evaluated the effects of B. etnensis Raf. bark methanolic extract on a human colon cancer cell line (CaCo2). In order to elucidate the mechanisms of action of the extract, cellular redox status, cell cycle, and heme oxygenase-1 (HO-1) expression in ferroptosis induction were evaluated. Cell viability and proliferation were tested by tetrazolium (MTT) assayand cell cycle analysis, while cell death was evaluated by annexin V test and lactic dehydrogenase (LDH) release. Cellular redox status was assessed by measuring thiol groups (RSH) content, reactive oxygen species (ROS) production, lipid hydroperoxide (LOOH) levels and (γ-glutamylcysteine synthetase) γ-GCS and HO-1 expressions. The extract significantly reduced cell viability of CaCo2, inducing necrotic cell death in a concentration-depending manner. In addition, an increase in ROS levels and a decrease of RSH content without modulation in γ-GCS expression were detected, with an augmentation in LOOH levels and drastic increase in HO-1 expression. These results suggest that the B. etnensis Raf. extract promotes an oxidative cellular microenvironment resulting in CaCo2 cell death by ferroptosis mediated by HO-1 hyper-expression.
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- 2019
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14. Radiation-Induced Gene Expression Changes in High and Low Grade Breast Cancer Cell Types
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Valentina Bravatà, Claudia Cava, Luigi Minafra, Francesco Paolo Cammarata, Giorgio Russo, Maria Carla Gilardi, Isabella Castiglioni, and Giusi Irma Forte
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ionizing radiation ,breast cancer ,gene expression profile ,pathway analysis ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Background: There is extensive scientific evidence that radiation therapy (RT) is a crucial treatment, either alone or in combination with other treatment modalities, for many types of cancer, including breast cancer (BC). BC is a heterogeneous disease at both clinical and molecular levels, presenting distinct subtypes linked to the hormone receptor (HR) status and associated with different clinical outcomes. The aim of this study was to assess the molecular changes induced by high doses of ionizing radiation (IR) on immortalized and primary BC cell lines grouped according to Human epidermal growth factor receptor (HER2), estrogen, and progesterone receptors, to study how HR status influences the radiation response. Our genomic approach using in vitro and ex-vivo models (e.g., primary cells) is a necessary first step for a translational study to describe the common driven radio-resistance features associated with HR status. This information will eventually allow clinicians to prescribe more personalized total doses or associated targeted therapies for specific tumor subtypes, thus enhancing cancer radio-sensitivity. Methods: Nontumorigenic (MCF10A) and BC (MCF7 and MDA-MB-231) immortalized cell lines, as well as healthy (HMEC) and BC (BCpc7 and BCpcEMT) primary cultures, were divided into low grade, high grade, and healthy groups according to their HR status. At 24 h post-treatment, the gene expression profiles induced by two doses of IR treatment with 9 and 23 Gy were analyzed by cDNA microarray technology to select and compare the differential gene and pathway expressions among the experimental groups. Results: We present a descriptive report of the substantial alterations in gene expression levels and pathways after IR treatment in both immortalized and primary cell cultures. Overall, the IR-induced gene expression profiles and pathways appear to be cell-line dependent. The data suggest that some specific gene and pathway signatures seem to be linked to HR status. Conclusions: Genomic biomarkers and gene-signatures of specific tumor subtypes, selected according to their HR status and molecular features, could facilitate personalized biological-driven RT treatment planning alone and in combination with targeted therapies.
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- 2018
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15. PET Images Atlas-Based Segmentation Performed in Native and in Template Space: A Radiomics Repeatability Study in Mouse Models.
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Paolo Giaccone, Viviana Benfante, Alessandro Stefano, Francesco Paolo Cammarata, Giorgio Russo, and Albert Comelli
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- 2022
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16. Area-based cell colony surviving fraction evaluation: A novel fully automatic approach using general-purpose acquisition hardware.
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Carmelo Militello, Leonardo Rundo, Vincenzo Conti, Luigi Minafra, Francesco Paolo Cammarata, Giancarlo Mauri, Maria Carla Gilardi, and Nunziatina Porcino
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- 2017
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17. The Role of Radiation in Cancer Treatment: New Insights towards Personalized Therapies
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Luigi Minafra, MARCO CALVARUSO, and Francesco Paolo Cammarata
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Medicine (miscellaneous) - Abstract
Despite all the recent pharmacological advances and the introduction of targeted therapies in clinical practice, cancer still remains one of the leading cause of death, accounting for 10 million deaths per year, based on the most recent reports [...]
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- 2022
18. Biological and Mechanical Characterization of the Random Positioning Machine (RPM) for Microgravity Simulations
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Valentina Bravatà, Veronica La Regina, Marco Calvaruso, Carmelo Militello, Luigi Minafra, Gaia Pucci, Giusi Irma Forte, Giorgio Ivan Russo, Filippo Torrisi, and Francesco Paolo Cammarata
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random positioning machine ,Space technology ,Random positioning machine ,Science ,Human life ,Paleontology ,space biology ,cancer biology ,TNBC ,simulated microgravity ,General Biochemistry, Genetics and Molecular Biology ,Space exploration ,Article ,simulatedmicrogravity ,randompositioningmachine ,Simulated microgravity ,Space and Planetary Science ,Systems engineering ,Cancer biology ,Interplanetary spaceflight ,Ecology, Evolution, Behavior and Systematics ,Space environment - Abstract
The rapid improvement of space technologies is leading to the continuous increase of space missions that will soon bring humans back to the Moon and, in the coming future, toward longer interplanetary missions such as the one to Mars. The idea of living in space is charming and fascinating; however, the space environment is a harsh place to host human life and exposes the crew to many physical challenges. The absence of gravity experienced in space affects many aspects of human biology and can be reproduced in vitro with the help of microgravity simulators. Simulated microgravity (s-μg) is applied in many fields of research, ranging from cell biology to physics, including cancer biology. In our study, we aimed to characterize, at the biological and mechanical level, a Random Positioning Machine in order to simulate microgravity in an in vitro model of Triple-Negative Breast Cancer (TNBC). We investigated the effects played by s-μg by analyzing the change of expression of some genes that drive proliferation, survival, cell death, cancer stemness, and metastasis in the human MDA-MB-231 cell line. Besides the mechanical verification of the RPM used in our studies, our biological findings highlighted the impact of s-μg and its putative involvement in cancer progression.
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- 2021
19. Radiosensitizing effect of curcumin-loaded lipid nanoparticles in breast cancer cells
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Giorgio Ivan Russo, Francesco Paolo Cammarata, Marcella Bonanomi, Daniela Gaglio, Carmelo Militello, Maria Carla Gilardi, Nunziatina Porcino, Margherita Baglio, G. Evangelista, Gaetano Savoca, B. Abbate, G. Iacoviello, Erika Amore, Giusi Irma Forte, Luigi Minafra, Valentina Bravatà, Maria Luisa Bondì, Minafra, L, Porcino, N, Bravatà, V, Gaglio, D, Bonanomi, M, Amore, E, Cammarata, F, Russo, G, Militello, C, Savoca, G, Baglio, M, Abbate, B, Iacoviello, G, Evangelista, G, Gilardi, M, Bondì, M, and Forte, G
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0301 basic medicine ,Radiation-Sensitizing Agents ,Radiosensitizer ,Curcumin ,Cell Survival ,medicine.medical_treatment ,lcsh:Medicine ,Antineoplastic Agents ,Apoptosis ,Breast Neoplasms ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,Drug Delivery Systems ,0302 clinical medicine ,Cell Line, Tumor ,medicine ,Humans ,Radiosensitivity ,Particle Size ,Clonogenic assay ,lcsh:Science ,IC50 ,Drug Carriers ,Multidisciplinary ,Radiotherapy ,Chemistry ,lcsh:R ,Radiotherapy, Systems biology ,Lipids ,In vitro ,Radiation therapy ,030104 developmental biology ,Cancer cell ,MCF-7 Cells ,Cancer research ,Nanoparticles ,Female ,lcsh:Q ,Systems biology ,030217 neurology & neurosurgery - Abstract
In breast cancer (BC) care, radiotherapy is considered an efficient treatment, prescribed both for controlling localized tumors or as a therapeutic option in case of inoperable, incompletely resected or recurrent tumors. However, approximately 90% of BC-related deaths are due to the metastatic tumor progression. Then, it is strongly desirable to improve tumor radiosensitivity using molecules with synergistic action. The main aim of this study is to develop curcumin-loaded solid nanoparticles (Cur-SLN) in order to increase curcumin bioavailability and to evaluate their radiosensitizing ability in comparison to free curcumin (free-Cur), by using an in vitro approach on BC cell lines. In addition, transcriptomic and metabolomic profiles, induced by Cur-SLN treatments, highlighted networks involved in this radiosensitization ability. The non tumorigenic MCF10A and the tumorigenic MCF7 and MDA-MB-231 BC cell lines were used. Curcumin-loaded solid nanoparticles were prepared using ethanolic precipitation and the loading capacity was evaluated by UV spectrophotometer analysis. Cell survival after treatments was evaluated by clonogenic assay. Dose–response curves were generated testing three concentrations of free-Cur and Cur-SLN in combination with increasing doses of IR (2–9 Gy). IC50 value and Dose Modifying Factor (DMF) was measured to quantify the sensitivity to curcumin and to combined treatments. A multi-“omic” approach was used to explain the Cur-SLN radiosensitizer effect by microarray and metobolomic analysis. We have shown the efficacy of the Cur-SLN formulation as radiosensitizer on three BC cell lines. The DMFs values, calculated at the isoeffect of SF = 50%, showed that the Luminal A MCF7 resulted sensitive to the combined treatments using increasing concentration of vehicled curcumin Cur-SLN (DMF: 1,78 with 10 µM Cur-SLN.) Instead, triple negative MDA-MB-231 cells were more sensitive to free-Cur, although these cells also receive a radiosensitization effect by combination with Cur-SLN (DMF: 1.38 with 10 µM Cur-SLN). The Cur-SLN radiosensitizing function, evaluated by transcriptomic and metabolomic approach, revealed anti-oxidant and anti-tumor effects. Curcumin loaded- SLN can be suggested in future preclinical and clinical studies to test its concomitant use during radiotherapy treatments with the double implications of being a radiosensitizing molecule against cancer cells, with a protective role against IR side effects.
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- 2019
20. Hypoxia Transcriptomic Modifications Induced by Proton Irradiation in U87 Glioblastoma Multiforme Cell Line
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Giusi Irma Forte, Francesco Paolo Cammarata, Olga Sokol, Walter Tinganelli, Giorgio Ivan Russo, Marco Calvaruso, Giada Petringa, G.A.P. Cirrone, Valentina Bravatà, Emanuele Scifoni, and Luigi Minafra
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0301 basic medicine ,Programmed cell death ,medicine.medical_treatment ,Medicine (miscellaneous) ,omic science ,Article ,Transcriptome ,03 medical and health sciences ,0302 clinical medicine ,Radioresistance ,medicine ,proton therapy ,transcriptome ,hypoxia ,glioblastoma ,ddc:610 ,U87 ,Chemistry ,Hypoxia (medical) ,Biomarker (cell) ,Radiation therapy ,030104 developmental biology ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,Medicine ,medicine.symptom - Abstract
Journal of Personalized Medicine 11(4), 308 (2021). doi:10.3390/jpm11040308, Published by MDPI, Basel
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- 2021
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21. Transcriptional modulations induced by proton irradiation in mice skin in function of adsorbed dose and distance
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Emiliano Fratini, Gihan Kamel, Giorgio Ivan Russo, Rosaria Acquaviva, Elena Di Nisio, Wei Wang, Francesco Paolo Cammarata, Roberto Amendola, Valerio Licursi, Lorenzo Manti, Mariangela Cestelli Guidi, Pietro Pisciotta, Rodolfo Negri, Licursi, Valerio, Wang, Wei, Di Nisio, Elena, Cammarata, Francesco P., Acquaviva, Rosaria, Russo, Giorgio, Manti, Lorenzo, Cestelli Guidi, Mariangela, Fratini, Emiliano, Kamel, Gihan, Amendola, Roberto, Pisciotta, Pietro, and Negri, Rodolfo
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Proton irradiation ,handrontherapy ,transcriptome modulation ,mouse skin ,spectroscopy FTIR microspectroscopy ,DIFFERENTIAL GENE-EXPRESSION ,integumentary system ,Proton ,IDENTIFICATION ,Chemistry ,Normal tissue ,ACUTE EXPOSURE ,RADIATION-INDUCED DAMAGE ,Ionizing radiation ,Hadron therapy ,Acute exposure ,Mouse skin ,CELLS ,Biophysics ,Irradiation ,IONIZING-RADIATION ,Function (biology) ,Proton irradiation, handrontherapy, transcriptome modulation, mouse skin, spectroscopy, FTIR microspectroscopy - Abstract
Hadron therapy by proton beams represents an advanced anti-cancer strategy due to its highly localized dose deposition allowing a greater sparing of normal tissue and/or organs at risk compared to photon/electron radiotherapy. However, it is not clear to what extent non-targeted effects such as transcriptional modulations produced along the beamline may diffuse and impact the surrounding tissue. In this work, we analyze the transcriptome of proton-irradiated mouse skin and choose two biomarker genes to trace their modulation at different distances from the beam's target and at different doses and times from irradiation to understand to what extent and how far it may propagate, using RNA-Seq and quantitative RT-PCR. In parallel, assessment of lipids alteration is performed by FTIR spectroscopy as a measure of tissue damage. Despite the observed high individual variability of expression, we can show evidence of transcriptional modulation of two biomarker genes at considerable distance from the beam's target where a simulation system predicts a significantly lower adsorbed dose. The results are compatible with a model involving diffusion of transcripts or regulatory molecules from high dose irradiated cells to distant tissue's portions adsorbing a much lower fraction of radiation.
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- 2021
22. Radiobiological outcomes, microdosimetric evaluations and monte carlo predictions in eye proton therapy
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Vladana Petković, Ivan Petrović, Valentina Bravatà, Marco Calvaruso, Giada Petringa, G.A.P. Cirrone, Francesco Paolo Cammarata, Lorenzo Manti, V. Conte, Luigi Minafra, Otilija Keta, Aleksandra M Ristić Fira, Selene Richiusa, Giacomo Cuttone, Giusi Irma Forte, Pavel Bláha, Giorgio Ivan Russo, Petringa, G., Calvaruso, M., Conte, V., Blaha, P., Bravata, V., Cammarata, F. P., Cuttone, G., Forte, G. I., Keta, O., Manti, L., Minafra, L., Petkovic, V., Petrovic, I., Richiusa, S., Fira, A. R., Russo, G., and Cirrone, G. A. P.
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Technology ,medicine.medical_specialty ,Radiobiology ,QH301-705.5 ,QC1-999 ,Monte Carlo method ,Sobp ,Normal tissue ,Geant4 ,RBE ,Microdosimetry ,030218 nuclear medicine & medical imaging ,03 medical and health sciences ,0302 clinical medicine ,medicine ,General Materials Science ,Medical physics ,Biology (General) ,protontherapy ,proton ,radiobiology ,microdosimetry ,QD1-999 ,Instrumentation ,Proton therapy ,Cell survival ,Fluid Flow and Transfer Processes ,Physics ,Protontherapy ,Process Chemistry and Technology ,General Engineering ,Experimental data ,Engineering (General). Civil engineering (General) ,3. Good health ,Computer Science Applications ,Chemistry ,030220 oncology & carcinogenesis ,Pigmented Epithelium ,Proton ,TA1-2040 - Abstract
CATANA (Centro di AdroTerapia ed Applicazioni Nucleari Avanzate) was the first Italian protontherapy facility dedicated to the treatment of ocular neoplastic pathologies. It is in operation at the LNS Laboratories of the Italian Institute for Nuclear Physics (INFN-LNS) and to date, 500 patients have been successfully treated. Even though proton therapy has demonstrated success in clinical settings, there is still a need for more accurate models because they are crucial for the estimation of clinically relevant RBE values. Since RBE can vary depending on several physical and biological parameters, there is a clear need for more experimental data to generate predictions. Establishing a database of cell survival experiments is therefore useful to accurately predict the effects of irradiations on both cancerous and normal tissue. The main aim of this work was to compare RBE values obtained from in-vitro experimental data with predictions made by the LEM II (Local Effect Model), Monte Carlo approaches, and semi-empirical models based on LET experimental measurements. For this purpose, the 92.1 uveal melanoma and ARPE-19 cells derived from normal retinal pigmented epithelium were selected and irradiated in the middle of clinical SOBP of the CATANA proton therapy facility. The remarkable results show the potentiality of using microdosimetric spectrum, Monte Carlo simulations and LEM model to predict not only the RBE but also the survival curves.
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- 2021
23. FT-IR transflection micro-spectroscopy study on normal human breast cells after exposure to a proton beam
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Giada Petringa, Maria Lasalvia, Marianna Portaccio, Ines Delfino, Maria Lepore, Giuseppe Perna, Francesco Paolo Cammarata, Vito Capozzi, Valerio Ricciardi, Lorenzo Manti, Pietro Pisciotta, Ricciardi, Valerio, Portaccio, Marianna, Perna, Giuseppe, Lasalvia, Maria, Capozzi, Vito, Cammarata, Francesco Paolo, Pisciotta, Pietro, Petringa, Giada, Delfino, Ine, Manti, Lorenzo, and Lepore, Maria
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Materials science ,fourier transform infrared micro-spectroscopy ,Proton ,Infrared ,02 engineering and technology ,fourier transform infrared micro-spectroscopy, transflection geometry, proton therapy, radiation dose effects, human breast cells ,Astrophysics::Cosmology and Extragalactic Astrophysics ,01 natural sciences ,lcsh:Technology ,Spectral line ,Ionizing radiation ,transflection geometry ,lcsh:Chemistry ,symbols.namesake ,Nuclear magnetic resonance ,proton therapy ,General Materials Science ,Irradiation ,Fourier transform infrared spectroscopy ,Instrumentation ,Proton therapy ,lcsh:QH301-705.5 ,radiation dose effects ,human breast cells ,Astrophysics::Galaxy Astrophysics ,Fluid Flow and Transfer Processes ,lcsh:T ,Process Chemistry and Technology ,010401 analytical chemistry ,radiation dose effect ,General Engineering ,021001 nanoscience & nanotechnology ,lcsh:QC1-999 ,0104 chemical sciences ,Computer Science Applications ,Fourier transform ,lcsh:Biology (General) ,lcsh:QD1-999 ,lcsh:TA1-2040 ,symbols ,Astrophysics::Earth and Planetary Astrophysics ,0210 nano-technology ,lcsh:Engineering (General). Civil engineering (General) ,lcsh:Physics - Abstract
Fourier transform infrared micro-spectroscopy (&mu, FT-IR) is nowadays considered a valuable tool for investigating the changes occurring in human cells after exposure to ionizing radiation. Recently, considerable attention has been devoted to the use of this optical technique in the study of cells exposed to proton beams, that are being increasingly adopted in cancer therapy. Different experimental configurations are used for proton irradiation and subsequent spectra acquisition. To facilitate the use of &mu, FT-IR, it may be useful to investigate new experimental approaches capable of speeding up and simplifying the irradiation and measurements phases. Here, we propose the use of low-e-substrates slides for cell culture, allowing the irradiation and spectra acquisition in transflection mode in a fast and direct way. In recent years, there has been a wide debate about the validity of these supports, but many researchers agree that the artifacts due to the presence of the electromagnetic standing wave effects are negligible in many practical cases. We investigated human normal breast cells (MCF-10 cell line) fixed immediately after the irradiation with graded proton radiation doses (0, 0.5, 2, and 4 Gy). The spectra obtained in transflection geometry showed characteristics very similar to those present in the spectra acquired in transmission geometry and confirm the validity of the chosen approach. The analysis of spectra indicates the occurrence of significant changes in DNA and lipids components of cells. Modifications in protein secondary structure are also evidenced.
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- 2021
24. Molecular Investigation on a Triple Negative Breast Cancer Xenograft Model Exposed to Proton Beams
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Giuseppe Broggi, Giacomo Cuttone, Marco Calvaruso, Giusi Irma Forte, Rosario Caltabiano, Giorgio Ivan Russo, Rosaria Acquaviva, Francesco Paolo Cammarata, Luigi Minafra, Giada Petringa, G.A.P. Cirrone, Valentina Bravatà, Filippo Torrisi, Roberta Tringali, Barbara Tomasello, and Pietro Pisciotta
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0301 basic medicine ,xenograft mice ,Apoptosis ,Triple Negative Breast Neoplasms ,THERAPY ,triple-negative breast cancer (TNBC) ,lcsh:Chemistry ,Mice ,0302 clinical medicine ,Cell Movement ,proton therapy ,Tumor Cells, Cultured ,Medicine ,lcsh:QH301-705.5 ,Spectroscopy ,Triple-negative breast cancer ,RISK ,Mice, Inbred BALB C ,General Medicine ,Cell cycle ,DOSE IONIZING-RADIATION ,Computer Science Applications ,Gene Expression Regulation, Neoplastic ,030220 oncology & carcinogenesis ,Immunohistochemistry ,Female ,Triple-negative breast cancer (TNBC) ,microarray ,Stem cell ,Protons ,medicine.drug_class ,Mice, Nude ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,Breast cancer ,In vivo ,MASTECTOMY ,Biomarkers, Tumor ,Animals ,Humans ,CELL ,Physical and Theoretical Chemistry ,Molecular Biology ,Cell Proliferation ,business.industry ,Gene Expression Profiling ,Organic Chemistry ,medicine.disease ,Xenograft Model Antitumor Assays ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Estrogen ,Molecular Response ,Cancer research ,business ,RESISTANCE - Abstract
Specific breast cancer (BC) subtypes are associated with bad prognoses due to the absence of successful treatment plans. The triple-negative breast cancer (TNBC) subtype, with estrogen (ER), progesterone (PR) and human epidermal growth factor-2 (HER2) negative receptor status, is a clinical challenge for oncologists, because of its aggressiveness and the absence of effective therapies. In addition, proton therapy (PT) represents an effective treatment against both inaccessible area located or conventional radiotherapy (RT)-resistant cancers, becoming a promising therapeutic choice for TNBC. Our study aimed to analyze the in vivo molecular response to PT and its efficacy in a MDA-MB-231 TNBC xenograft model. TNBC xenograft models were irradiated with 2, 6 and 9 Gy of PT. Gene expression profile (GEP) analyses and immunohistochemical assay (IHC) were performed to highlight specific pathways and key molecules involved in cell response to the radiation. GEP analysis revealed in depth the molecular response to PT, showing a considerable immune response, cell cycle and stem cell process regulation. Only the dose of 9 Gy shifted the balance toward pro-death signaling as a dose escalation which can be easily performed using proton beams, which permit targeting tumors while avoiding damage to the surrounding healthy tissue.
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- 2020
25. The role of hypoxia and SRC tyrosine kinase in glioblastoma invasiveness and radioresistance
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Gaetano Magro, Federica M. Spitale, Rosario Gulino, Lucia Salvatorelli, Giacomo Cuttone, Filippo Torrisi, Rosalba Parenti, Luigi Minafra, Giorgio Ivan Russo, Nunzio Vicario, Francesco Paolo Cammarata, Samuel Valable, Department of Biomedical and Biotechnological Sciences [Catania, Italy], University of Catania [Italy], Institute of Molecular Bioimaging and Physiology, National Research Council, Università degli studi di Catania [Catania], Istituto Nazionale di Fisica Nucleare, Sezione di Catania (INFN), Imagerie et Stratégies Thérapeutiques des pathologies Cérébrales et Tumorales (ISTCT), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Laboratori Nazionali del Sud (LNS), Istituto Nazionale di Fisica Nucleare (INFN), and Università degli studi di Catania = University of Catania (Unict)
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0301 basic medicine ,Cancer Research ,medicine.medical_treatment ,[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology ,Brain tumor ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Review ,lcsh:RC254-282 ,Targeted therapy ,03 medical and health sciences ,0302 clinical medicine ,SRC tyrosine kinase ,Radioresistance ,medicine ,business.industry ,hypoxia ,Hypoxia (medical) ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,medicine.disease ,invasion ,targeted therapy ,Radiation therapy ,radioresistance ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,medicine.symptom ,business ,Glioblastoma ,Tyrosine kinase ,Proto-oncogene tyrosine-protein kinase Src - Abstract
Simple Summary The biological pathways underlying glioblastoma malignancy and radioresistance are still unclear. In this review, we describe the role of the hypoxic microenvironment and SRC proto-oncogene non-receptor tyrosine kinase in the activation of radioresistance and invasion pathways of glioblastoma. We also highlight the hypoxia- and ionizing radiation-induced infiltration, providing updated evidences on the involvement of SRC in these processes. Optimizing radiotherapy and identifying druggable molecular players are crucial steps to improve current glioblastoma therapeutic strategies. Abstract Advances in functional imaging are supporting neurosurgery and radiotherapy for glioblastoma, which still remains the most aggressive brain tumor with poor prognosis. The typical infiltration pattern of glioblastoma, which impedes a complete surgical resection, is coupled with a high rate of invasiveness and radioresistance, thus further limiting efficient therapy, leading to inevitable and fatal recurrences. Hypoxia is of crucial importance in gliomagenesis and, besides reducing radiotherapy efficacy, also induces cellular and molecular mediators that foster proliferation and invasion. In this review, we aimed at analyzing the biological mechanism of glioblastoma invasiveness and radioresistance in hypoxic niches of glioblastoma. We also discussed the link between hypoxia and radiation-induced radioresistance with activation of SRC proto-oncogene non-receptor tyrosine kinase, prospecting potential strategies to overcome the current limitation in glioblastoma treatment.
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- 2020
26. MF2C3: Multi-feature fuzzy clustering to enhance cell colony detection in automated clonogenic assay evaluation
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Vincenzo Conti, Carmelo Militello, Marco Calvaruso, Francesco Paolo Cammarata, Giorgio Ivan Russo, Leonardo Rundo, Luigi Minafra, Rundo, Leonardo [0000-0003-3341-5483], Apollo - University of Cambridge Repository, Militello, Carmelo [0000-0003-2249-9538], Minafra, Luigi [0000-0003-3112-4519], Cammarata, Francesco Paolo [0000-0002-0554-6649], Calvaruso, Marco [0000-0003-2387-8273], Conti, Vincenzo [0000-0002-8718-111X], and Russo, Giorgio [0000-0003-1493-1087]
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Fuzzy clustering ,Physics and Astronomy (miscellaneous) ,Computer science ,General Mathematics ,02 engineering and technology ,clonogenic assay ,automatic cell colony detection ,spatial fuzzy c-means clustering ,multi-feature clustering ,entropy ,standard deviation ,Fuzzy logic ,Standard deviation ,03 medical and health sciences ,0202 electrical engineering, electronic engineering, information engineering ,Computer Science (miscellaneous) ,Entropy (information theory) ,Cluster analysis ,Clonogenic assay ,030304 developmental biology ,0303 health sciences ,Pixel ,business.industry ,lcsh:Mathematics ,Pattern recognition ,lcsh:QA1-939 ,Thresholding ,Automatic cell colony detection ,Entropy ,Multi-feature clustering ,Spatial fuzzy c-means clustering ,Chemistry (miscellaneous) ,020201 artificial intelligence & image processing ,Artificial intelligence ,business - Abstract
© 2020 by the authors. A clonogenic assay is a biological technique for calculating the Surviving Fraction (SF) that quantifies the anti-proliferative effect of treatments on cell cultures: this evaluation is often performed via manual counting of cell colony-forming units. Unfortunately, this procedure is error-prone and strongly affected by operator dependence. Besides, conventional assessment does not deal with the colony size, which is generally correlated with the delivered radiation dose or administered cytotoxic agent. Relying upon the direct proportional relationship between the Area Covered by Colony (ACC) and the colony count and size, along with the growth rate, we propose MF2C3, a novel computational method leveraging spatial Fuzzy C-Means clustering on multiple local features (i.e., entropy and standard deviation extracted from the input color images acquired by a general-purpose flat-bed scanner) for ACC-based SF quantification, by considering only the covering percentage. To evaluate the accuracy of the proposed fully automatic approach, we compared the SFs obtained by MF2C3 against the conventional counting procedure on four different cell lines. The achieved results revealed a high correlation with the ground-truth measurements based on colony counting, by outperforming our previously validated method using local thresholding on L*u*v* color well images. In conclusion, the proposed multi-feature approach, which inherently leverages the concept of symmetry in the pixel local distributions, might be reliably used in biological studies.
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- 2020
27. DNA damage assessment of human breast and lung carcinoma cells irradiated with protons and carbon ions
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Vladana Petković, Ivan Petrović, Otilija Keta, Giada Petringa, Aleksandra Ristić-Fira, P. Cirrone, Giacomo Cuttone, Sebastien Incerti, Francesco Paolo Cammarata, Centre d'Etudes Nucléaires de Bordeaux Gradignan (CENBG), and Université Sciences et Technologies - Bordeaux 1-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)
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DNA damage ,chemistry.chemical_element ,010403 inorganic & nuclear chemistry ,DNA damage response ,01 natural sciences ,030218 nuclear medicine & medical imaging ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,Carcinoma ,skin and connective tissue diseases ,Lung cancer ,Lung ,Chemistry ,medicine.disease ,Scavenger (chemistry) ,0104 chemical sciences ,carbon ions ,medicine.anatomical_structure ,Cancer cell ,Cancer research ,[PHYS.PHYS.PHYS-MED-PH]Physics [physics]/Physics [physics]/Medical Physics [physics.med-ph] ,cancer cells ,Hydroxyl radical ,Protons ,Carbon - Abstract
International audience; Differences in biological response of breast and lung cancer cells to 62 MeV/u therapeutic protons and carbon ions are investigated on MCF-7 and HTB177 cells. Hydroxyl radical scavenger, dimethyl sulfoxide (DMSO) is applied to reduce indirect effects of irradiation, while serum deprivation provided uniformity of cell population. Survival, immunocytochemical, and cell-cycle analysis, changes in protein expression involved in repair and apoptosis are followed. Radiobiological parameters, Bax/Bcl-2 ratio, and increased subG1 fraction show that carbon ions are more efficient in cellular killing than protons. No significant difference in the number of γH2AX foci are found between protons and carbon ions. According to SF2 values, MCF-7 cells are more radioresistant to both ion species compared to HTB177 cells. Cell-cycle analysis and expression of relevant protein markers further confirm somewhat higher radiosensitivity of HTB177 cells compared to MCF-7. DMSO leads to the rise of cell survival and decreases the number of γH2AX foci. Even though there are no significant changes in the number of γH2AX foci after used irradiation types, lesions induced by carbon ions are probably more complex than those produced by protons. DMSO minimizes indirect DNA damage to achieve experimental conditions needed for comparisons of obtained results with numerical simulations
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- 2020
28. Nutraceutical Compounds as Sensitizers for Cancer Treatment in Radiation Therapy
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Giorgio Ivan Russo, Francesco Paolo Cammarata, Valentina Bravatà, Gaia Pucci, Marco Calvaruso, Luigi Minafra, Giusi Irma Forte, and Rosa Musso
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Oncology ,medicine.medical_specialty ,Radiation-Sensitizing Agents ,medicine.medical_treatment ,Review ,Catalysis ,Inorganic Chemistry ,lcsh:Chemistry ,Nutraceutical ,Internal medicine ,Neoplasms ,medicine ,cancer ,Animals ,Humans ,Radiosensitivity ,Physical and Theoretical Chemistry ,Molecular Biology ,Cell damage ,lcsh:QH301-705.5 ,Spectroscopy ,nutraceuticals ,Chemotherapy ,Radiotherapy ,business.industry ,Organic Chemistry ,Cancer ,General Medicine ,medicine.disease ,radiotherapy ,Computer Science Applications ,Cancer treatment ,Radiation therapy ,lcsh:Biology (General) ,lcsh:QD1-999 ,Cancer cell ,Dietary Supplements ,business - Abstract
The improvement of diagnostic techniques and the efficacy of new therapies in clinical practice have allowed cancer patients to reach a higher chance to be cured together with a better quality of life. However, tumors still represent the second leading cause of death worldwide. On the contrary, chemotherapy and radiotherapy (RT) still lack treatment plans which take into account the biological features of tumors and depend on this for their response to treatment. Tumor cells’ response to RT is strictly-connected to their radiosensitivity, namely, their ability to resist and to overcome cell damage induced by ionizing radiation (IR). For this reason, radiobiological research is focusing on the ability of chemical compounds to radiosensitize cancer cells so to make them more responsive to IR. In recent years, the interests of researchers have been focused on natural compounds that show antitumoral effects with limited collateral issues. Moreover, nutraceuticals are easy to recover and are thus less expensive. On these bases, several scientific projects have aimed to test also their ability to induce tumor radiosensitization both in vitro and in vivo. The goal of this review is to describe what is known about the role of nutraceuticals in radiotherapy, their use and their potential application.
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- 2019
29. Gene Expression Profiles Induced by High-dose Ionizing Radiation in MDA-MB-231 Triple-negative Breast Cancer Cell Line
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Valentina Bravatà, Giorgio Ivan Russo, Ivan Fazio, Gaia Pucci, Giusi Irma Forte, Luigi Minafra, Rosa Musso, Massimiliano Spada, and Francesco Paolo Cammarata
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Cancer Research ,medicine.medical_treatment ,Triple Negative Breast Neoplasms ,Biology ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,Ionizing radiation (IR) ,MDA-MB-231 cells ,gene expression profile (GEP) ,Radioresistance ,Cell Line, Tumor ,Radiation, Ionizing ,Gene expression ,Genetics ,medicine ,Humans ,Clonogenic assay ,Molecular Biology ,Triple-negative breast cancer ,Microarray analysis techniques ,medicine.disease ,Radiation therapy ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,Female ,Transcriptome ,Research Article - Abstract
Background/aim Radiation therapy (RT) represents a therapeutic option in breast cancer (BC). Even if a great number of BC patients receive RT, not all of them report benefits, due to radioresistance that gets activated through several factors, such as the hormone receptor status. Herein, we analyzed the gene expression profiles (GEP) induced by RT in triple-negative BC (TNBC) MDA-MB-231, to study signalling networks involved in radioresistance. Materials and methods GEP of MDA-MB-231 BC cells treated with a high dose of radiation, went through cDNA microarray analysis. In addition, to examine the cellular effects induced by RT, analyses of morphology and clonogenic evaluation were also conducted. Results A descriptive report of GEP and pathways induced by IR is reported from our microarray data. Moreover, the MDA-MB-231 Radioresistent Cell Fraction (RCF) selected, included specific molecules able to drive radioresistance. Conclusion In summary, our data highlight, the RT response of TNBC MDA-MB-231 cell line at a transcriptional level, in terms of activating radioresistance in these cells, as a model of late-stage BC.
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- 2019
30. Raman spectroscopy for the evaluation of the radiobiological sensitivity of normal human breast cells at different time points after irradiation by a clinical proton beam
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Maria Lasalvia 1, 2, Giuseppe Perna 1, Pietro Pisciotta 3, 4, Francesco Paolo Cammarata 4, Lorenzo Manti 5, 6, Vito Capozzi 1, Lasalvia, M., Perna, G., Pisciotta, P., Cammarata, F. P., Manti, L., and Capozzi, V.
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Time Factors ,Proton ,Time Factor ,medicine.medical_treatment ,Apoptosis ,02 engineering and technology ,Spectrum Analysis, Raman ,01 natural sciences ,Biochemistry ,Analytical Chemistry ,symbols.namesake ,Necrosis ,Breast cancer ,Electrochemistry ,medicine ,Environmental Chemistry ,Humans ,Irradiation ,Breast ,Clonogenic assay ,Proton therapy ,Spectroscopy ,Cells, Cultured ,Cellular Senescence ,Micronuclei, Chromosome-Defective ,Cell Proliferation ,Chemistry ,Raman spectroscopy ,breast cells ,010401 analytical chemistry ,Apoptosi ,Dose-Response Relationship, Radiation ,021001 nanoscience & nanotechnology ,medicine.disease ,Necrosi ,In vitro ,0104 chemical sciences ,Radiation therapy ,symbols ,Biophysics ,Female ,Protons ,0210 nano-technology ,Human ,DNA Damage - Abstract
Among different radiotherapy techniques, proton irradiation is an established and effective method for treatment of several types of cancer, because less healthy tissue is exposed with respect to conventional radiotherapy by photons/electrons. Recently, proton therapy has been proposed for the treatment of breast cancer. In vitro studies of proton irradiated normal human breast cells can provide information about cellular radioresponse, particularly as far as healthy tissue is concerned. In this paper, a study of the effects at different time points, following proton irradiation at different doses, of human normal MCF10A breast cells is performed by Raman spectroscopy. The aim of this investigation is to detect the unwanted effects of proton treatment and to investigate the possibility of monitoring them and of making an assessment of the cellular sensitivity by means of such a technique. The obtained results seem to indicate a rather significant sensitivity of MCF10A cells to proton irradiation. In fact, even at doses as low as 0.5 Gy, biological effects are clearly detectable in Raman spectra. In particular, ratiometric analysis of the Raman spectra measured from the nucleoplasm compartment showed that DNA/RNA damage increases with time, suggesting that most cells are unable to repair DNA/RNA broken bonds. The results obtained by the Raman spectroscopy analysis exhibit a similar trend with regard to dose to those obtained by commonly used radiobiological assays (i.e. MTT, clonogenic assay, senescence, apoptosis and necrosis). The results of this study strongly suggest the possibility that the Raman technique can be used to identify molecular markers predicting radiation response.
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- 2019
31. Betula etnensis Raf. (Betulaceae) Extract Induced HO-1 Expression and Ferroptosis Cell Death in Human Colon Cancer Cells
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Giuseppe Antonio Malfa 1, Barbara Tomasello 1, Rosaria Acquaviva 1, Carlo Genovese 2, Alfonsina La Mantia 1, Francesco Paolo Cammarata 3, 4, Monica Ragusa 5, Marcella Renis 1, and Claudia Di Giacomo 1
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0301 basic medicine ,Programmed cell death ,Betula etnensis Raf ,Oxidative phosphorylation ,medicine.disease_cause ,Article ,Catalysis ,Colon cancer ,oxidative stress ,heme oxigenase-1 ,ferroptosis ,thiol groups ,lcsh:Chemistry ,Inorganic Chemistry ,03 medical and health sciences ,0302 clinical medicine ,Annexin ,medicine ,Viability assay ,Physical and Theoretical Chemistry ,lcsh:QH301-705.5 ,Molecular Biology ,Spectroscopy ,chemistry.chemical_classification ,Reactive oxygen species ,Organic Chemistry ,General Medicine ,Cell cycle ,Molecular biology ,Computer Science Applications ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,chemistry ,Cell culture ,030220 oncology & carcinogenesis ,Oxidative stress - Abstract
Betula etnensis Raf. (Birch Etna) belonging to the Betulaceae family grows on the eastern slope of Etna. Many bioactive compounds present in Betula species are considered promising anticancer agents. In this study, we evaluated the effects of B. etnensis Raf. bark methanolic extract on a human colon cancer cell line (CaCo2). In order to elucidate the mechanisms of action of the extract, cellular redox status, cell cycle, and heme oxygenase-1 (HO-1) expression in ferroptosis induction were evaluated. Cell viability and proliferation were tested by tetrazolium (MTT) assayand cell cycle analysis, while cell death was evaluated by annexin V test and lactic dehydrogenase (LDH) release. Cellular redox status was assessed by measuring thiol groups (RSH) content, reactive oxygen species (ROS) production, lipid hydroperoxide (LOOH) levels and (&gamma, glutamylcysteine synthetase) &gamma, GCS and HO-1 expressions. The extract significantly reduced cell viability of CaCo2, inducing necrotic cell death in a concentration-depending manner. In addition, an increase in ROS levels and a decrease of RSH content without modulation in &gamma, GCS expression were detected, with an augmentation in LOOH levels and drastic increase in HO-1 expression. These results suggest that the B. etnensis Raf. extract promotes an oxidative cellular microenvironment resulting in CaCo2 cell death by ferroptosis mediated by HO-1 hyper-expression.
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- 2019
32. Radiobiological quantities in proton-therapy: Estimation and validation using Geant4-based Monte Carlo simulations
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J. Pipek, Andrea Attili, Francesco Romano, G. Forte, Giacomo Cuttone, P. Pisciotta, Giada Petringa, G.A.P. Cirrone, Luciano Pandola, Francesco Paolo Cammarata, L Manganaro, Lorenzo Manti, G. V. Russo, Petringa, G., Romano, F., Manti, L., Pandola, L., Attili, A., Cammarata, F., Cuttone, G., Forte, G., Manganaro, L., Pipek, J., Pisciotta, P., Russo, G., and Cirrone, G. A. P.
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Survival fraction ,LOCAL EFFECT MODEL ,Proton ,CELL-SURVIVAL ,Physics::Instrumentation and Detectors ,Monte Carlo method ,Physics::Medical Physics ,Biophysics ,Reproducibility of Result ,General Physics and Astronomy ,Geant4 ,RBE ,BEAM ,030218 nuclear medicine & medical imaging ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Proton Therapy ,Radiobiological model ,Dosimetry ,Humans ,Radiology, Nuclear Medicine and imaging ,HEAVY-ION RADIOTHERAPY ,Proton therapy ,Monte Carlo ,Coupling ,Physics ,REPAIR ,Physics::Computational Physics ,Radiotherapy ,Experimental data ,Radiobiology ,Reproducibility of Results ,Radiotherapy Dosage ,General Medicine ,IN-VITRO ,Computational physics ,030220 oncology & carcinogenesis ,Monte Carlo Method ,Human - Abstract
Purpose The Geant4 Monte Carlo simulation toolkit was used to reproduce radiobiological parameters measured by irradiating three different cancerous cell lines with monochromatic and clinical proton beams. Methods The experimental set-up adopted for irradiations was fully simulated with a dedicated open-source Geant4 application. Cells survival fractions was calculated coupling the Geant4 simulations with two analytical radiobiological models: one based on the LEM (Local Effect Model) approach and the other on a semi-empirical parameterisation. Results was evaluated and compared with experimental data. Results and conclusions The results demonstrated the Geant4 ability to reproduce radiobiological quantities for different cell lines.
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- 2019
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33. Specific protein changes contribute to the differential muscle mass loss during ageing
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Cecilia Gelfi, Michele Vasso, Patrizia Procacci, Sara De Palma, Daniele Capitanio, Enrica Torretta, Francesco Paolo Cammarata, Chiara Fania, Valerio Magnaghi, Capitanio, D, Vasso, M, De Palma, S, Fania, C, Torretta, E, Cammarata, F, Magnaghi, V, Procacci, P, and Gelfi, C
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Male ,Proteomics ,0301 basic medicine ,Aging ,Respiratory chain ,Muscle Proteins ,Hindlimb ,Muscle Protein ,Muscle proteome ,Biology ,Biochemistry ,Rats, Sprague-Dawley ,Two-Dimensional Difference Gel Electrophoresis ,Muscle ageing ,03 medical and health sciences ,Gastrocnemius muscle ,chemistry.chemical_compound ,Muscular Diseases ,Myosin ,Autophagy ,medicine ,Animals ,Muscle, Skeletal ,Molecular Biology ,Tissue homeostasis ,Myosin Heavy Chains ,Mass spectrometry ,Animal ,Muscular Disease ,Myosin Heavy Chain ,Two-Dimensional Difference Gel Electrophoresi ,Proteomic ,Skeletal muscle ,Animal proteomics ,Intermediate metabolism ,Mitochondria ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Myoglobin ,chemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,2D-DIGE ,Animal proteomic ,Forelimb - Abstract
In the skeletal muscle, the ageing process is characterized by a loss of muscle mass and strength, coupled with a decline of mitochondrial function and a decrease of satellite cells. This profile is more pronounced in hindlimb than in forelimb muscles, both in humans and in rodents. Utilizing light and electron microscopy, myosin heavy chain isoform distribution, proteomic analysis by 2D-DIGE, MALDI-TOF MS and quantitative immunoblotting, this study analyzes the protein levels and the nuclear localization of specific molecules, which can contribute to a preferential muscle loss. Our results identify the molecular changes in the hindlimb (gastrocnemius) and forelimb (triceps) muscles during ageing in rats (3- and 22-month-old). Specifically, the oxidative metabolism contributes to tissue homeostasis in triceps, whereas respiratory chain disruption and oxidative-stress-induced damage imbalance the homeostasis in gastrocnemius muscle. High levels of dihydrolipoyllysine-residue acetyltransferase (Dlat) and ATP synthase subunit alpha (Atp5a1) are detected in triceps and gastrocnemius, respectively. Interestingly, in triceps, both molecules are increased in the nucleus in aged rats and are associated to an increased protein acetylation and myoglobin availability. Furthermore, autophagy is retained in triceps whereas an enhanced fusion, decrement of mitophagy and of regenerative potential is observed in aged gastrocnemius muscle.
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- 2016
34. Radiation-Induced Gene Expression Changes in High and Low Grade Breast Cancer Cell Types
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Isabella Castiglioni, Claudia Cava, Francesco Paolo Cammarata, Giorgio Ivan Russo, Maria Carla Gilardi, Giusi Irma Forte, Luigi Minafra, Valentina Bravatà, Bravata, V, Cava, C, Minafra, L, Cammarata, F, Russo, G, Gilardi, M, Castiglioni, I, and Forte, G
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0301 basic medicine ,medicine.medical_treatment ,ionizing radiation ,breast cancer ,gene expression profile ,pathway analysis ,Radiation Tolerance ,lcsh:Chemistry ,0302 clinical medicine ,Radiation, Ionizing ,Gene expression ,Medicine ,Gene Regulatory Networks ,Receptor ,lcsh:QH301-705.5 ,Spectroscopy ,Cell Line, Transformed ,General Medicine ,3. Good health ,Computer Science Applications ,Gene Expression Regulation, Neoplastic ,Hormone receptor ,030220 oncology & carcinogenesis ,Female ,Breast cancer, Gene expression profile, Ionizing radiation, Pathway analysis ,Signal Transduction ,Breast Neoplasms ,Radiation Dosage ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,Breast cancer ,Cell Line, Tumor ,Humans ,Physical and Theoretical Chemistry ,Molecular Biology ,business.industry ,Gene Expression Profiling ,Organic Chemistry ,Computational Biology ,Cancer ,Dose-Response Relationship, Radiation ,Radiation-Induced Gene Expression ,medicine.disease ,Radiation therapy ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Cancer research ,Gene chip analysis ,Neoplasm Grading ,Transcriptome ,business - Abstract
Background: There is extensive scientific evidence that radiation therapy (RT) is a crucial treatment, either alone or in combination with other treatment modalities, for many types of cancer, including breast cancer (BC). BC is a heterogeneous disease at both clinical and molecular levels, presenting distinct subtypes linked to the hormone receptor (HR) status and associated with different clinical outcomes. The aim of this study was to assess the molecular changes induced by high doses of ionizing radiation (IR) on immortalized and primary BC cell lines grouped according to Human epidermal growth factor receptor (HER2), estrogen, and progesterone receptors, to study how HR status influences the radiation response. Our genomic approach using in vitro and ex-vivo models (e.g., primary cells) is a necessary first step for a translational study to describe the common driven radio-resistance features associated with HR status. This information will eventually allow clinicians to prescribe more personalized total doses or associated targeted therapies for specific tumor subtypes, thus enhancing cancer radio-sensitivity. Methods: Nontumorigenic (MCF10A) and BC (MCF7 and MDA-MB-231) immortalized cell lines, as well as healthy (HMEC) and BC (BCpc7 and BCpcEMT) primary cultures, were divided into low grade, high grade, and healthy groups according to their HR status. At 24 h post-treatment, the gene expression profiles induced by two doses of IR treatment with 9 and 23 Gy were analyzed by cDNA microarray technology to select and compare the differential gene and pathway expressions among the experimental groups. Results: We present a descriptive report of the substantial alterations in gene expression levels and pathways after IR treatment in both immortalized and primary cell cultures. Overall, the IR-induced gene expression profiles and pathways appear to be cell-line dependent. The data suggest that some specific gene and pathway signatures seem to be linked to HR status. Conclusions: Genomic biomarkers and gene-signatures of specific tumor subtypes, selected according to their HR status and molecular features, could facilitate personalized biological-driven RT treatment planning alone and in combination with targeted therapies.
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- 2018
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35. Radiogenomics: the utility in patient selection
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Pietro Pisciotta, Giusi Irma Forte, Valentina Bravatà, Maria Carla Gilardi, Luigi Minafra, G.A.P. Cirrone, Giorgio Ivan Russo, Debora Lamia, Francesco Paolo Cammarata, Giacomo Cuttone, Forte, G, Minafra, L, Bravata, V, Cammarata, F, Lamia, D, Pisciotta, P, Cirrone, G, Cuttone, G, Gilardi, M, and Russo, G
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0301 basic medicine ,Oncology ,Cancer Research ,medicine.medical_specialty ,medicine.medical_treatment ,Radiogenomics ,Tumor heterogeneity ,03 medical and health sciences ,0302 clinical medicine ,Therapeutic index ,Cancer stem cell ,Internal medicine ,Omics, Radiation therapy (RT), Radiation toxicity ,medicine ,Radiology, Nuclear Medicine and imaging ,In patient ,Medical physics ,Radiation treatment planning ,business.industry ,Radiation therapy ,030104 developmental biology ,030220 oncology & carcinogenesis ,State of art ,business ,Radiation therapy (RT) ,radiation toxicity ,omics - Abstract
The goal of radiation therapy (RT) is to deliver the therapeutic dose to target tissues minimizing the risks of normal tissue complication. Nowadays, technological advances in radiation delivery and the introduction of particle therapies have strongly limited the amount of dose distributed to normal tissues and enhanced the tumor killing capacity. Here, we discuss about the state of art in RT treatment modalities, tumor sensitivity and radiation toxicity. A special insight is dedicated to the role of tumor heterogeneity, cancer stem cells (CSCs) and hypoxia. In addition, in this review we provide an overview of the most recently studies evaluating the potential role of “omic” biomarkers for a personalized biological-driven treatment planning, useful to maximize the radiotherapy success without normal tissues complications.
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- 2017
36. High-Intensity Focused Ultrasound- and Radiation Therapy-Induced Immuno-modulation: Comparison and Potential Opportunities
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Roberta Cirincione 1, Federica Maria Di Maggio 1, 2, Giusi Irma Forte 1, Luigi Minafra 1, Valentina Bravatà 1, Laura Castiglia 1, Vincenzo Cavalieri 4, Giovanni Borasi 1, Giorgio Russo 1, Domenico Lio 2, Cristina Messa 1, 4, 5, Maria Carla Gilardi 1, Francesco Paolo Cammarata 1, Cirincione, R, Di Maggio, F, Forte, G, Minafra, L, Bravatà, V, Castiglia, L, Cavalieri, V, Borasi, G, Russo, G, Lio, D, Messa, C, Gilardi, M, Cammarata, F, and Messa, M
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,Radiology, Nuclear Medicine and Imaging ,Acoustics and Ultrasonics ,medicine.medical_treatment ,Biophysics ,Immunomodulation ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Neoplasms ,medicine ,Bystander effect ,Humans ,Cancer ,Chemotherapy ,Radiological and Ultrasound Technology ,business.industry ,medicine.disease ,Primary tumor ,Thermal ablation ,High-intensity focused ultrasound ,Radiation therapy ,Immuno-therapy ,Tumor vaccine ,030104 developmental biology ,Biophysic ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,High-Intensity Focused Ultrasound Ablation ,Anti-tumor immunity ,business - Abstract
In recent years, high-intensity focused ultrasound (HIFU) has emerged as a new and promising non-invasive and non-ionizing ablative technique for the treatment of localized solid tumors. Extensive pre-clinical and clinical studies have evidenced that, in addition to direct destruction of the primary tumor, HIFU-thermoablation may elicit long-term systemic host anti-tumor immunity. In particular, an important consequence of HIFU treatment includes the release of tumor-associated antigens (TAAs), the secretion of immuno-suppressing factors by cancer cells and the induction of cytotoxic T lymphocyte (CTL) activity. Radiation therapy (RT) is the main treatment modality used for many types of tumors and about 50% of all cancer patients receive RT, often used in combination with surgery and chemotherapy. It is well known that RT can modulate anti-tumor immune responses, modifying micro-environment and stimulating inflammatory factors that can greatly affect cell invasion, bystander effects, radiation tissue complications (such as fibrosis), genomic instability and thus, intrinsic cellular radio-sensitivity. To date, various combined therapeutic strategies (such as immuno-therapy) have been performed in order to enhance RT success in treating locally advanced and recurrent tumors. Recent works suggested the combined use of HIFU and RT treatments to increase the tumor cell radio-sensitivity, in order to synergize the effects reaching the maximum results with minimal doses of ionizing radiation (IR). Here, we highlight the opposite immuno-modulation roles of RT and HIFU, providing scientific reasons to test, by experimental approaches, the use of HIFU immune-stimulatory capacity to improve tumor radio-sensitivity, to reduce the RT induced inflammatory response and to decrease the dose-correlated side effects in normal tissues.
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- 2017
37. Preliminary study for small animal preclinical hadrontherapy facility
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Valentina Bravatà, Giusi Irma Forte, G.A.P. Cirrone, V. Marchese, Luigi Minafra, Francesco Paolo Cammarata, Pietro Pisciotta, Rosaria Acquaviva, Debora Lamia, Giacomo Cuttone, Fabrizio Romano, G. V. Russo, Maria Carla Gilardi, Russo, G, Pisciotta, P, Cirrone, G, Romano, F, Cammarata, F, Marchese, V, Forte, G, Lamia, D, Minafra, L, Bravata, V, Acquaviva, R, Gilardi, M, and Cuttone, G
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Physics ,Nuclear and High Energy Physics ,medicine.medical_specialty ,Monte Carlo method ,Detector ,Preclinical studies ,Geant4 ,Imaging phantom ,030218 nuclear medicine & medical imaging ,Gamma index ,03 medical and health sciences ,DICOM ,0302 clinical medicine ,030220 oncology & carcinogenesis ,Small animal ,Hadrontherapy ,Dosimetry ,Small animal irradiation protocol ,medicine ,Medical physics ,Tumour configuration ,Dosimetry, Geant4, Hadrontherapy, Preclinical studies, Small animal irradiation protocol ,Instrumentation ,Simulation - Abstract
Aim of this work is the study of the preliminary steps to perform a particle treatment of cancer cells inoculated in small animals and to realize a preclinical hadrontherapy facility. A well-defined dosimetric protocol was developed to explicate the steps needed in order to perform a precise proton irradiation in small animals and achieve a highly conformal dose into the target. A precise homemade positioning and holding system for small animals was designed and developed at INFN-LNS in Catania (Italy), where an accurate Monte Carlo simulation was developed, using Geant4 code to simulate the treatment in order to choose the best animal position and perform accurately all the necessary dosimetric evaluations. The Geant4 application can also be used to realize dosimetric studies and its peculiarity consists in the possibility to introduce the real target composition in the simulation using the DICOM micro-CT image. This application was fully validated comparing the results with the experimental measurements. The latter ones were performed at the CATANA (Centro di AdroTerapia e Applicazioni Nucleari Avanzate) facility at INFN-LNS by irradiating both PMMA and water solid phantom. Dosimetric measurements were performed using previously calibrated EBT3 Gafchromic films as a detector and the results were compared with the Geant4 simulation ones. In particular, two different types of dosimetric studies were performed: the first one involved irradiation of a phantom made up of water solid slabs where a layer of EBT3 was alternated with two different slabs in a sandwich configuration, in order to validate the dosimetric distribution. The second one involved irradiation of a PMMA phantom made up of a half hemisphere and some PMMA slabs in order to simulate a subcutaneous tumour configuration, normally used in preclinical studies. In order to evaluate the accordance between experimental and simulation results, two different statistical tests were made: Kolmogorov test and gamma index test. This work represents the first step towards the realization of a preclinical hadrontherapy facility at INFN-LNS in Catania for the future in vivo studies.
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- 2017
38. Using Delayed luminescence to evaluate radiobiological risks
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Grasso, Rosaria, FRANCESCO PAOLO CAMMARATA, Luigi, Minafra, Valentina, Marchese, Giorgio, Russo, Lorenzo, Manti, Musumeci, Francesco, and Scordino, Agata
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- 2017
39. Cytokine profile of breast cell lines after different radiation doses
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Francesco Paolo Cammarata, Maria Carla Gilardi, Valentina Bravatà, Federica Maria Di Maggio, Giusi Irma Forte, Domenico Lio, Luigi Minafra, Giorgio Ivan Russo, Giuseppa Augello, Bravatà, Valentina, Minafra, Luigi, Forte, Giusi Irma, Cammarata, Francesco Paolo, Russo, Giorgio, Di Maggio, Federica Maria, Augello, Giuseppa, Lio, Domenico, Gilardi, Maria Carla, Bravata, V, Minafra, L, Forte, G, Cammarata, F, Russo, G, Di Maggio, F, Augello, G, Lio, D, and Gilardi, M
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0301 basic medicine ,Ionizing radiation ,Radiology, Nuclear Medicine and Imaging ,Cell Survival ,Cytokine profile ,Breast Neoplasms ,Inflammation ,Radiation ,Radiation Tolerance ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,breast cancer ,Cell Line, Tumor ,medicine ,cytokine ,Humans ,skin and connective tissue diseases ,Cell survival ,Radiological and Ultrasound Technology ,Chemistry ,breast cancer, cytokines, inflammation, Ionizing radiation, Breast Neoplasms, Cell Line, Tumor, Cell Survival, Culture Media, Conditioned, Dose-Response Relationship, Radiation, Humans, Phenotype, Radiation Tolerance ,Dose-Response Relationship, Radiation ,medicine.disease ,cytokines ,Dose–response relationship ,030104 developmental biology ,Phenotype ,Cell culture ,inflammation ,030220 oncology & carcinogenesis ,Culture Media, Conditioned ,Immunology ,Cancer research ,medicine.symptom ,Breast Neoplasm ,Human - Abstract
Purpose: Ionizing radiation (IR) treatment activates inflammatory processes causing the release of a great amount of molecules able to affect the cell survival. The aim of this study was to analyze the cytokine signature of conditioned medium produced by non-tumorigenic mammary epithelial cell line MCF10A, as well as MCF7 and MDA-MB-231 breast cancer cell lines, after single high doses of IR in order to understand their role in high radiation response. Materials and methods: We performed a cytokine profile of irradiated conditioned media of MCF10A, MCF7 and MDA-MB-231 cell lines treated with 9 or 23 Gy, by Luminex and ELISA analyses. Results: Overall, our results show that both 9 Gy and 23 Gy of IR induce the release within the first 72 h of cytokines and growth factors potentially able to influence the tumor outcome, with a dose-independent and cell-line dependent signature. Moreover, our results show that the cell-senescence phenomenon does not correlate with the amount of ‘senescence-associated secretory phenotype’ (SASP) molecules released in media. Thus, additional mechanisms are probably involved in this process. Conclusions: These data open the possibility to evaluate cytokine profile as useful marker in modulating the personalized radiotherapy in breast cancer care.
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- 2017
40. Area-based cell colony surviving fraction evaluation: A novel fully automatic approach using general-purpose acquisition hardware
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Carmelo Militello a, Leonardo Rundo a, b, Vincenzo Conti c, Luigi Minafra a, Francesco Paolo Cammarata a, Giancarlo Mauri b, Maria Carla Gilardi a, Nunziatina Porcino a, Militello, C, Rundo, L, Conti, V, Minafra, L, Cammarata, F, Mauri, G, Gilardi, M, and Porcino, N
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0301 basic medicine ,ING-INF/06 - BIOINGEGNERIA ELETTRONICA E INFORMATICA ,Clonogenic assays ,Computer science ,Cell Survival ,Cell Culture Techniques ,Health Informatics ,Local adaptive thresholding ,Cell Count ,Breast Neoplasms ,02 engineering and technology ,ING-INF/05 - SISTEMI DI ELABORAZIONE DELLE INFORMAZIONI ,Hough transform ,law.invention ,Circle Hough transform ,03 medical and health sciences ,law ,Area covered by colony ,0202 electrical engineering, electronic engineering, information engineering ,Humans ,Fraction (mathematics) ,Clonogenic assay ,Delivered radiation dose ,Colony-forming unit ,business.industry ,INF/01 - INFORMATICA ,Pattern recognition ,Thresholding ,Computer Science Applications ,Fully automatic surviving fraction evaluation ,General-purpose acquisition hardware ,Female ,MCF-7 Cells ,Neoplastic Stem Cells ,030104 developmental biology ,General purpose ,Fully automatic ,020201 artificial intelligence & image processing ,Artificial intelligence ,business - Abstract
Background The current methodology for the Surviving Fraction (SF) measurement in clonogenic assay, which is a technique to study the anti-proliferative effect of treatments on cell cultures, involves manual counting of cell colony forming units. This procedure is operator-dependent and error-prone. Moreover, the identification of the exact colony number is often not feasible due to the high growth rate leading to the adjacent colony merging. As a matter of fact, conventional assessment does not deal with the colony size, which is generally correlated with the delivered radiation dose or the administered cytotoxic agent. Method Considering that the Area Covered by Colony (ACC) is proportional to the colony number and size as well as to the growth rate, we propose a novel fully automatic approach exploiting Circle Hough Transform, to automatically detect the wells in the plate, and local adaptive thresholding, which calculates the percentage of ACC for the SF quantification. This measurement relies just on this covering percentage and does not consider the colony number, preventing inconsistencies due to intra- and inter-operator variability. Results To evaluate the accuracy of the proposed approach, we compared the SFs obtained by our automatic ACC-based method against the conventional counting procedure. The achieved results (r = 0.9791 and r = 0.9682 on MCF7 and MCF10A cells, respectively) showed values highly correlated with the measurements using the traditional approach based on colony number alone. Conclusions The proposed computer-assisted methodology could be integrated in laboratory practice as an expert system for the SF evaluation in clonogenic assays.
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- 2017
41. Delayed luminescence in a multiparameter approach to evaluation and reduction of radiobiological risks [NOVEL BIOPHOTONICS TECHNIQUES AND APPLICATIONS IV]
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Grasso, Rosaria, Francesco Paolo Cammarata, Luigi, Minafra, Valentina, Marchese, Giorgio, Russo, Lorenzo, Manti, Musumeci, Francesco, and Scordino, Agata
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delayed luminescence, non-invasive technique, non-tumorigenic, MCF10A, proton beam, ion irradiation ,MCF10A ,ion irradiation ,proton beam ,non-tumorigenic ,non-invasive technique ,delayed luminescence - Published
- 2017
42. 49. Combined treatments with Hadrontherapy – in vitro tests and preclinical approach
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Giorgio Ivan Russo, Luigi Minafra, Valentina Bravatà, Pietro Pisciotta, Filippo Torrisi, Giovanna Calabrese, Francesco Paolo Cammarata, Laura Maccari, Giusi Irma Forte, Anna Lucia Fallacara, V. Marchese, Maurizio Botta, Rosalba Parenti, Giacomo Cuttone, and G.A.P. Cirrone
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Oncology ,medicine.medical_specialty ,business.industry ,Biophysics ,General Physics and Astronomy ,General Medicine ,medicine.disease ,In vitro ,Hadron therapy ,Therapy response ,Internal medicine ,medicine ,Radiology, Nuclear Medicine and imaging ,Molecular imaging ,U87 ,business ,Dose rate ,Proton therapy ,Glioblastoma - Abstract
Purpose Glioblastoma multiforme (GBM) is the most aggressive types of central nervous system tumors. The aim of the project is to validate an innovative molecule developed by Lead Discovery Siena [LDS] showing inhibitory activity against Src kinases to be associated with proton therapy treatments. Materials Human glioblastoma cells (U87) were treated at INFN-LNS in Catania with 2 and 10 Gy of proton beams at dose rate of 10 Gy/min in the middle of the extended modulated Bragg peak and in combination with different concentrations of the LDS molecule (10 and 20 μM). The fraction of surviving cells (SF) was evaluated with the clonogenicity test 15 days after irradiation. Results The data in the figure show the synergistic effect of the combined treatment with LDS. Conclusions In vitro studies confirmed the efficacy of the hadron therapy treatment in combination with the molecule. The next step is to translate these studies on the mouse model thanks to the existence of an Animal Facility created by the synergy between the following institution: the LNS-INFN, the University of Catania – Capir Center, IBFM-CNR and H. Cannizzaro, with the departments of Nuclear Medicine and Medical Physics. The facility will be able to provide unique research activities in the European panorama: hadron therapy treatments, dose distribution studies using the Monte Carlo GEANT4 method, molecular imaging using microPET/ CT tomography with different radiotracers and image processing implementation for therapy response quantification.
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- 2018
43. Radiation Therapy Towards Laser-Driven Particle Beams: An 'OMICS' Approach in Radiobiology
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Francesco Paolo Cammarata, Giusi Irma Forte, Luigi Minafra, and Valentina Bravatà
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0301 basic medicine ,Physics ,medicine.medical_specialty ,Radiobiology ,medicine.medical_treatment ,Nanotechnology ,Proteomics ,Laser ,Tumor control ,law.invention ,Ionizing radiation ,Radiation therapy ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,law ,030220 oncology & carcinogenesis ,Molecular Response ,Relative biological effectiveness ,medicine ,Medical physics - Abstract
The main goal of radiation therapy (RT) treatments is to achieve local tumor control, to selectively kill cancer cells without causing significant damage to the surrounding normal tissues. RT uses ionizing radiation (IR) generated with conventional accelerators, such as X-rays, γ-rays, electrons, protons and ions. It is now well recognized by the entire scientific community that to evaluate the biological effects of IR it is essential an “OMIC” approach to take into account both the different cell types involved and the different IR’s used. The latest advances on cell and molecular response to IR, the most relevant data emerging from recent studies (genomics, epigenetics, proteomics and immunology) about different cell types, will be reported. We will discuss mainly biological effects of IR generated by conventional accelerators but we will also consider the few and preliminary radiobiological studies performed so far with laser-driven electron and proton beams. This will allow us in particular to speculate on cell and molecular effects of the laser-driven electron beams, a topic that can be now carefully investigated thanks to the impressive progress of “table-top” laser-driven accelerators.
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- 2016
44. Gene expression profiling of breast cancer cell lines treated with proton and electron radiations
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Francesco Paolo Cammarata, Maria Carla Gilardi, Giada Petringa, Debora Lamia, Valentina Bravatà, G.A.P. Cirrone, V. Marchese, Luigi Minafra, Giacomo Cuttone, Lorenzo Manti, Pietro Pisciotta, Giorgio Ivan Russo, Giusi Irma Forte, Bravata, V., Minafra, L., Cammarata, F. P., Pisciotta, P., Lamia, D., Marchese, V., Petringa, G., Manti, L., Cirrone, G. A. P., Gilardi, M. C., Cuttone, G., Forte, G. I., Russo, G., Bravata, V, Minafra, L, Cammarata, F, Pisciotta, P, Lamia, D, Marchese, V, Petringa, G, Manti, L, Cirrone, G, Gilardi, M, Cuttone, G, Forte, G, and Russo, G
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0301 basic medicine ,Proton ,Quantitative Biology::Tissues and Organs ,medicine.medical_treatment ,Nuclear Theory ,Physics::Medical Physics ,Gene Expression ,Breast Neoplasms ,Electrons ,Electron ,Adenocarcinoma ,Radiation Tolerance ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,Conventional radiotherapy ,Breast cancer cell line ,Adenocarcinoma, Breast Neoplasms, Carcinoma, Ductal, Breast, Cell Line, Cell Line, Tumor, Electrons, Gene Expression, Humans, Linear Energy Transfer, Oligonucleotide Array Sequence Analysis, Precision Medicine, Radiation Tolerance, Gene Expression Profiling, Proton Therapy ,Cell Line, Tumor ,Proton Therapy ,medicine ,Humans ,Linear Energy Transfer ,Radiology, Nuclear Medicine and imaging ,Precision Medicine ,Nuclear Experiment ,Oligonucleotide Array Sequence Analysis ,FACTOR-KAPPA-B ,TRANSCRIPTION FACTOR ,INDUCED APOPTOSIS ,ENERGY-TRANSFER ,MONTE-CARLO ,THERAPY ,RADIOTHERAPY ,SIMULATION ,INHIBITION ,PATHWAY ,Full Paper ,Chemistry ,Gene Expression Profiling ,Carcinoma, Ductal, Breast ,General Medicine ,Gene expression profiling ,Radiation therapy ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research - Abstract
OBJECTIVE: Technological advances in radiation therapy are evolving with the use of hadrons, such as protons, indicated for tumors where conventional radiotherapy does not give significant advantages or for tumors located in sensitive regions, which need the maximum of dose-saving of the surrounding healthy tissues. The genomic response to conventional and non conventional Linear Energy Transfer exposure is a poor investigated topic and became an issue of radiobiological interest. The aim of this work was to analyze and compare molecular responses in term of gene expression profiles, induced by electron and proton irradiation in breast cancer cell lines. METHODS: We studied the gene expression profiling differences by cDNA microarray activated in response to electron and proton irradiation with different Linear Energy Transfer values, among three breast cell lines (the tumorigenic MCF7 and MDA-MB-231 and the non tumorigenic MCF10A), exposed to the same sub-lethal dose of 9 Gy. RESULTS: Gene expression profiling pathway analyses showed the activation of different signaling and molecular networks in a cell line and radiation type-dependent manner. MCF10A and MDA-MB-231 cell lines were found to induce factors and pathways involved in the immunological process control. CONCLUSIONS: Here we describe in a detailed way the gene expression profiling and pathways activated after electron and proton irradiation in breast cancer cells. Summarizing, although specific pathways are activated in a radiation type-dependent manner, each cell line activates overall similar molecular networks in response to both these two types of ionizing radiation. Advances in knowledge: In the era of personalized medicine and breast cancer target-directed intervention, we trust that this study could drive radiation therapy towards personalized treatments, evaluating possible combined treatments, based on the molecular characterization.
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- 2018
45. Caveolin-1, breast cancer and ionizing radiation
- Author
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Pucci, M., Bravatà, V., Forte, G. I., Francesco Paolo Cammarata, Messa, C., Gilardi, M. C., Minafra, L., Pucci, M., Bravatà, V., Forte, G., Cammarata, F., Messa, C., Gilardi, M., Minafra, L., Pucci, M, Bravatà, V, Forte, G, Cammarata, F, Messa, M, Gilardi, M, and Minafra, L
- Subjects
Radiotherapy ,Animal ,CAV1 ,biomarker ,breast cancer ,ionizing radiation ,review ,Animals ,Biomarkers, Tumor ,Breast Neoplasms ,Caveolin 1 ,Cell Transformation, Neoplastic ,Female ,Gene Expression Regulation, Neoplastic ,Humans ,Molecular Targeted Therapy ,Receptor, Epidermal Growth Factor ,Radiation, Ionizing ,ErbB Receptors ,Breast Neoplasm ,Human - Abstract
Breast cancer (BC) recovery has increased in recent years thanks to efforts of Omics-based research in this field. However, despite the important results obtained, BC remains a complex multifactorial pathology that is difficult to treat appropriately. Caveolin-1 (CAV1), the basic constituent protein of specialized plasma membrane invaginations called caveolae, is emerging as a potential therapeutic biomarker in BC. This factor may modulate BC response to chemotherapy and radiation therapy. In addition, recent reports describe the key role of CAV1 during cell response to oxidative stress. The aim of the present review was to describe the biological roles of CAV1 in BC considering its contrasting dual functions as an oncogene and as a tumor suppressor. In addition, we report on how CAV1 may contribute to tumor cell response to ionizing radiation treatment. Finally, new roles of CAV1 in BC both on epithelium and stroma may be useful as prognostic indicators for patient treatment and help clinicians in the selection of the best personalized therapy.
- Published
- 2015
46. High-dose Ionizing Radiation Regulates Gene Expression Changes in the MCF7 Breast Cancer Cell Line
- Author
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Valentina Bravatà 1, Luigi Minafra 1, Giorgio Russo 1, Giusi Irma Forte 1, Francesco Paolo Cammarata 1, Marilena Ripamonti 1, Carlo Casarino 1, Giuseppa Augello 2, Francesca Costantini 2, Giovanna Barbieri 2, Cristina Messa 1, 3, 5, Maria Carla Gilardi 1, and 4
- Subjects
MCF7 breast cancer cells ,Intraoperative electron radiation therapy ,IR ,microarray ,ionizing radiation ,IOERT - Abstract
BACKGROUND: Intraoperative electron radiation therapy (IOERT) is a therapeutic technique which administers a single high dose of ionizing radiation immediately after surgical tumor removal. IOERT induces a strong stress response: both tumor and normal cells activating pro- and antiproliferative cell signaling pathways. Following treatment, several genes and factors are differently modulated, producing an imbalance in cell fate decision. However, the contribution of these genes and pathways in conferring different cell radiosensitivity and radioresistance needs to be further investigated, in particular after high-dose treatments. Despite the documented and great impact of IOERT in breast cancer care, and the trend for dose escalation, very limited data are available regarding gene-expression profiles and cell networks activated by IOERT or high-dose treatment. The aim of the study was to analyze the main pathways activated following high radiation doses in order to select for potential new biomarkers of radiosensitivity or radioresistance, as well as to identify therapeutic targets useful in cancer care. MATERIALS AND METHODS: We performed gene-expression profiling of the MCF7 human breast carcinoma cell line after treatment with 9- and 23-Gy doses (conventionally used during IOERT boost and exclusive treatments, respectively) by cDNA microarrays. Real-Time Quantitative Reverse Transcription PCR (qRT-PCR), immunofluorescence and immunoblot experiments were performed to validate candidate IOERT biomarkers. We also conducted clonogenic tests and cellular senescence assays to monitor for radiation-induced effects. RESULTS: The analyses highlighted a transcriptome dependent on the dose delivered and a number of specific key genes that may be proposed as new markers of radiosensitivity. Cell and molecular traits observed in MCF7 cells revealed a typical senescent phenotype associated with cell proliferation arrest after treatments with 9- and 23-Gy doses. CONCLUSION: In this study, we report genes and cellular networks activated following high-dose IOERT. The selected validated genes were used to design two descriptive models for each dose delivered. We believe that this study could contribute to the understanding over the complex mechanisms which regulate cell radiosensitivity and radioresistance in order to improve personalized radiotherapeutic treatment.
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- 2015
47. Abstract ID: 35 Monte Carlo dosimetric study for preclinical small animal hadrontherapy using Geant4 toolkit
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Pietro Pisciotta, Giusi Irma Forte, Francesco Paolo Cammarata, Rosaria Acquaviva, Giorgio Ivan Russo, Debora Lamia, Francesco Romano, Giacomo Cuttone, Giada Petringa, G.A.P. Cirrone, and Maria Carla Gilardi
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Positioning system ,Computer science ,Monte Carlo method ,Detector ,Biophysics ,General Physics and Astronomy ,Experimental data ,General Medicine ,Imaging phantom ,030218 nuclear medicine & medical imaging ,Gamma index ,03 medical and health sciences ,DICOM ,0302 clinical medicine ,030220 oncology & carcinogenesis ,Small animal ,Radiology, Nuclear Medicine and imaging ,Simulation ,Biomedical engineering - Abstract
Preclinical studies represent an important step to study molecular mechanism induced inside cells in response to ionizing radiations. The scope was the study of the preliminary steps to perform particle treatment of cancer cells inoculated in small animals and to realize a preclinical hadrontherapy facility. At this scope, a well-defined dosimetric protocol was developed to perform the steps needed in order to perform a precise proton irradiation in small animals and achieve highly conformal dose. Homemade positioning system for small animals was developed at INFN-LNS(Italy) and an accurate Monte Carlo simulation was developed. The application, developed using Geant4.10.03 version, includes in the current version of the advanced example “Hadrontherapy” [1] the main functionalities of the “DICOM” extended example [2] . In this way, this application simulates the CATANA beam line geometry and includes the capability to implement DICOM-TC images as target. The application will be used to carry out dosimetric and LET studies [3] using the real target composition. This application was validated comparing its results with experimental measurements. A validation test, using solid water slabs phantom, and a treatment simulation, irradiating a PMMA phantom that simulate subcutaneous tumours, were executed at CATANA facility using in both cases EBT3-Gafchromic films as dose detector. The dosimetric results were compared with the simulation ones using Kolmogorov and gamma index test. Experimental data are in good agreement with the simulation ones. The phantom targets were included in the simulation environment using DICOM-TC images. Dosimetric measurements were useful to determine the efficiency of the developed Geant4 application and to demonstrate that it is a valid instrument to study the dose distribution in different types of phantoms with different kinds of geometry. This work has been carried-out in the perspective of realization of a preclinical hadrontherapy facility at INFN-LNS in order to implement interesting future in vivo studies using small animals.
- Published
- 2017
48. Gene expression profiling of epithelial-mesenchymal transition in primary breast cancer cell culture
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Luigi, Minafra, Valentina, Bravatà, Giusi Irma, Forte, Francesco Paolo, Cammarata, Maria Carla, Gilardi, and Cristina, Messa
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Epithelial-Mesenchymal Transition ,Cell Movement ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Carcinoma, Ductal, Breast ,Humans ,Breast Neoplasms ,Female ,Real-Time Polymerase Chain Reaction ,Transcriptome ,Cells, Cultured ,Oligonucleotide Array Sequence Analysis - Abstract
Epithelial-mesenchymal transition (EMT) is a process co-opted by cancer cells to invade and form metastases. In the present study we analyzed gene expression profiles of primary breast cancer cells in culture in order to highlight genes related to EMT.Microarray expression analysis of primary cells isolated from a specimen of a patient with an infiltrating ductal carcinoma of the breast was performed. Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) analyses validated microarray gene expression trends.Thirty-six candidate genes were selected and used to generate a molecular network displaying the tight relationship among them. The most significant Gene Ontology biological processes characterizing this network were involved in cell migration and motility.Our data revealed the involvement of new genes which displayed tight relationships among them, suggesting a molecular network in which they could contribute to control of EMT in breast cancer. This study may offer a basis for understanding complex mechanisms which regulate breast cancer progression and for designing individualized anticancer therapies.
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- 2014
49. 'Omics' of HER2-positive breast cancer
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Francesco Paolo Cammarata, Valentina Bravatà, Luigi Minafra, Giusi Irma Forte, Bravatà, V, Cammarata, FP, Forte, GI, and Minafra, L
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Proteomics ,Receptor, ErbB-2 ,Breast Neoplasms ,Biology ,Bioinformatics ,Antibodies, Monoclonal, Humanized ,Biochemistry ,Epigenesis, Genetic ,HER2/Neu Positive ,Germline mutation ,Breast cancer ,breast cancer ,Trastuzumab ,Genetics ,medicine ,Biomarkers, Tumor ,Humans ,OMICS ,Epigenetics ,skin and connective tissue diseases ,neoplasms ,Molecular Biology ,Genomics ,Genes, erbB-2 ,Omics ,medicine.disease ,Tumor progression ,Molecular Medicine ,HER2/neu-positive ,Female ,Transcriptome ,Biotechnology ,medicine.drug - Abstract
HER2/neu amplification/overexpression is the only somatic mutation widely considered to be a marker of disease outcome and response to treatment in breast cancer. Pathologists have made large efforts to achieve accuracy in characterizing HER2/neu status. The introduction of transtuzumab contributed to development of additional measures to identify sensitive and resistant subclasses of HER2/neu-positive tumors. In this article, we describe the latest advances in HER2/neu status diagnostic assessment and the most relevant research emerging from ‘‘Omics’’ (genomics, epigenetics, transcriptomics, and proteomics) studies on HER2/neu-positive breast cancer. A large quantity of biomarkers from different studies highlighted HER2/neu-positive specific proliferation, cell cycle arrest, and apoptosis mechanisms, as well as immunological and metabolic behavior. Major driver genes of tumor progression have had a candidate status (GRB7, MYC, CCND1, EGFR, etc.), even though the main role for HER2/neu is largely recognized. Nonetheless, existing omics data and HER2/neupositive molecular profiles seem to suggest that few proteogenomic alterations in HER2, EGFR, and PI3K networks could significantly affect the effectiveness of transtuzumab. The systematic search of molecular alterations in and across these pathways can help to select the most appropriate drug for a given patient based on in-depth understanding of complexity in tumor biology.
- Published
- 2013
50. Single-nucleotide polymorphisms and FDG-PET in breast cancer
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Valentina Bravatà, Alessandro Stefano, Francesco Paolo Cammarata, Luigi Minafra, Giorgio Russo, S. Nicolosi, Sabina Pulizzi, V. Tripoli, Isabella Castiglioni, Cecilia Gelfi, Maria Carla Gilardi, and Messa Cristina.
- Published
- 2013
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