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1. Gold Coated Nanoparticles Functionalized by Photochemical Immobilization Technique for Immunosensing

Author

Della Ventura, B., Campanile, R., Cimafonte, M., Elia, V., Forente, A., Minopoli, A., Scardapane, E., Schiattarella, C., Velotta, R., Angrisani, Leopoldo, Series Editor, Arteaga, Marco, Series Editor, Panigrahi, Bijaya Ketan, Series Editor, Chakraborty, Samarjit, Series Editor, Chen, Jiming, Series Editor, Chen, Shanben, Series Editor, Chen, Tan Kay, Series Editor, Dillmann, Rüdiger, Series Editor, Duan, Haibin, Series Editor, Ferrari, Gianluigi, Series Editor, Ferre, Manuel, Series Editor, Hirche, Sandra, Series Editor, Jabbari, Faryar, Series Editor, Jia, Limin, Series Editor, Kacprzyk, Janusz, Series Editor, Khamis, Alaa, Series Editor, Kroeger, Torsten, Series Editor, Liang, Qilian, Series Editor, Martín, Ferran, Series Editor, Ming, Tan Cher, Series Editor, Minker, Wolfgang, Series Editor, Misra, Pradeep, Series Editor, Möller, Sebastian, Series Editor, Mukhopadhyay, Subhas, Series Editor, Ning, Cun-Zheng, Series Editor, Nishida, Toyoaki, Series Editor, Pascucci, Federica, Series Editor, Qin, Yong, Series Editor, Seng, Gan Woon, Series Editor, Speidel, Joachim, Series Editor, Veiga, Germano, Series Editor, Wu, Haitao, Series Editor, Zhang, Junjie James, Series Editor, Di Francia, Girolamo, editor, and Di Natale, Corrado, editor

Published
2021
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2. The Nanomechanical Properties of CLL Cells Are Linked to the Actin Cytoskeleton and Are a Potential Target of BTK Inhibitors

Author

Sampietro, M, Cassina, V, Salerno, D, Barbaglio, F, Buglione, E, Marrano, C, Campanile, R, Scarfò, L, Biedenweg, D, Fregin, B, Zamai, M, Díaz Torres, A, Labrador Cantarero, V, Ghia, P, Otto, O, Mantegazza, F, Caiolfa, V, Scielzo, C, Sampietro M., Cassina V., Salerno D., Barbaglio F., Buglione E., Marrano C. A., Campanile R., Scarfò L., Biedenweg D., Fregin B., Zamai M., Díaz Torres A., Labrador Cantarero V., Ghia P., Otto O., Mantegazza F., Caiolfa V. R., Scielzo C., Sampietro, M, Cassina, V, Salerno, D, Barbaglio, F, Buglione, E, Marrano, C, Campanile, R, Scarfò, L, Biedenweg, D, Fregin, B, Zamai, M, Díaz Torres, A, Labrador Cantarero, V, Ghia, P, Otto, O, Mantegazza, F, Caiolfa, V, Scielzo, C, Sampietro M., Cassina V., Salerno D., Barbaglio F., Buglione E., Marrano C. A., Campanile R., Scarfò L., Biedenweg D., Fregin B., Zamai M., Díaz Torres A., Labrador Cantarero V., Ghia P., Otto O., Mantegazza F., Caiolfa V. R., and Scielzo C.

Abstract

Chronic lymphocytic leukemia (CLL) is an incurable disease characterized by an intense trafficking of the leukemic cells between the peripheral blood and lymphoid tissues. It is known that the ability of lymphocytes to recirculate strongly depends on their capability to rapidly rearrange their cytoskeleton and adapt to external cues; however, little is known about the differences occurring between CLL and healthy B cells during these processes. To investigate this point, we applied a single-cell optical (super resolution microscopy) and nanomechanical approaches (atomic force microscopy, real-time deformability cytometry) to both CLL and healthy B lymphocytes and compared their behavior. We demonstrated that CLL cells have a specific actomyosin complex organization and altered mechanical properties in comparison to their healthy counterpart. To evaluate the clinical relevance of our findings, we treated the cells in vitro with the Bruton's tyrosine kinase inhibitors and we found for the first time that the drug restores the CLL cells mechanical properties to a healthy phenotype and activates the actomyosin complex. We further validated these results in vivo on CLL cells isolated from patients undergoing ibrutinib treatment. Our results suggest that CLL cells' mechanical properties are linked to their actin cytoskeleton organization and might be involved in novel mechanisms of drug resistance, thus becoming a new potential therapeutic target aiming at the normalization of the mechanical fingerprints of the leukemic cells.

Published
2023

3. Nanoscale analysis reveals distinct mechanical properties in Chronic Lymphocytic Leukemia cells

Author

Campanile, R and Campanile, R

Abstract

Chronic lymphocytic leukemia (CLL) is the most frequent leukemia in the Western world, with an incidence of 5 new cases per 100.000 individuals annually. The disease affects mainly elderly people, however one third of the new cases are diagnosed before the age of 55 years. CLL is an incurable ailment characterised by an intense trafficking and accumulation of mature CD5+ B lymphocytes in peripheral blood, bone marrow (BM) and secondary lymphoid organs (spleen and lymph nodes). It is known that the ability of lymphocytes to recirculate strongly depends on their capability to rapidly rearrange their cytoskeleton and adapt to external cues; however, little is known about the differences occurring between CLL and healthy B (HB) cells during these processes. To investigate this point, we performed a comparison between primary healthy, leukemic B cells by applying nanomechanical approaches (Atomic Force Microscopy). By means of single cells optical super resolution microscopy we demonstrated that CLL cells have a specific actomyosin complex organization and altered mechanical properties in comparison to their healthy counterpart. To evaluate the clinical relevance of our findings, we treated the cells in vitro with the Bruton’s tyrosine kinase inhibitors and we found for the first time that the drug restores the CLL cells mechanical properties to a healthy phenotype and activates the actomyosin complex. We further validated these results in vivo on CLL cells isolated from patients undergoing ibrutinib treatment. Our results suggest that CLL cells’ mechanical properties are linked to their actin cytoskeleton organization and might be involved in novel mechanisms of drug resistance, thus becoming a new potential therapeutic target aiming at the normalization of the mechanical fingerprints of the leukemic cells. Furthermore, we employed representative cellular lines of CLL (MEC Wild-Type) cells, which are indicative of the disease characteristics. Additionally, we generated

Published
2023

4. Nanoscale analysis reveals distinct cytoskeletal architecture and mechanical properties in Chronic Lymphocytic Leukemia cells

Author

Campanile, R and Campanile, R

Abstract

Chronic lymphocytic leukemia (CLL) is the most frequent leukemia in the Western world, with an incidence of 5 new cases per 100.000 individuals annually. The disease affects mainly elderly people, however one third of the new cases are diagnosed before the age of 55 years. CLL is an incurable ailment characterised by an intense trafficking and accumulation of mature CD5+ B lymphocytes in peripheral blood, bone marrow (BM) and secondary lymphoid organs (spleen and lymph nodes). It is known that the ability of lymphocytes to recirculate strongly depends on their capability to rapidly rearrange their cytoskeleton and adapt to external cues, however little is known about the differences occurring between CLL and healthy B cells (HB) in this context. Particularly, HB cells can reorganize completely their architecture; an ability depending on their structural plasticity, which facilitates B cells migration and communicative function. To investigate this point, we performed a comparison between primary healthy, leukemic B cells by applying single cells optical (super resolution microscopy), and nanomechanical approaches (Atomic Force Microscopy, Real Time Deformability Cytometry). We demonstrated that CLL cells have a specific nanoscale actomyosin complex organisation and altered mechanical properties in comparison to their healthy counterpart. To evaluate the clinical relevance of our findings we treated the cells in vitro with the BTK inhibitor ibrutinib and we found for the first time that the drug restores the CLL cells mechanical properties towards the healthy phenotype and activates the actomyosin complex in a BCR independent manner. We further validated these results in vivo on CLL cells isolated from patients undergoing ibrutinib treatment. Considering that the majority of CLL patients develop a resistance to ibrutinib, we analysed a resistant case, and we observed a return to the pre-treatment properties. Our results suggest that CLL cells' mechanical properties a

Published
2023

21. Drought tolerance of photosynthetic electron transport under CO2-enriched and normal air in cereal species.

Author

Flagella, Z., Campanile, R. G., Stoppelli, M. C., De Caro, A., and Di Fonzo, N.

Subjects
*ELECTRON transport, *PHOTOSYNTHESIS, *CEREAL grasses, *DROUGHTS, *BARLEY, *FLUORIMETRY, *BIOCHEMISTRY, *METALLOENZYMES
Abstract

The quantum yield of photosynthetic electron transport (ΦPSII), evaluated by means of chlorophyll (Chl) fluorescence analysis, has proven to be a useful screening test for drought tolerance in durum wheat (Triticum durum Desf.). To explore the potential of this parameter further in detecting drought-tolerant genotypes, three cereal species were studied; ΦPSII measurements were carried out under two different gas mixtures, at three points of the induction curve (to obtain the maximal ΦPSII and both the transient and steady-state actual ΦPSII), and at three different water stress levels (moderate, severe and drastic). The species investigated were durum and bread wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.); two cultivars per species, characterized by different levels of drought tolerance, were tested. The two gas mixtures used were normal air (21% O2, 0.035% CO2 in N2) to monitor the whole photosynthetic process under physiological conditions, and CO2 enriched-low O2 air (1% O2, 5% CO2 in N2) to monitor ΦPSII reduction under stress mainly related to Calvin cycle activity. When ΦPSII related to both assimilatory and non-assimilatory metabolism was evaluated, the cultivar differences observed under normal Air were more representative of the agronomic performance upon drought stress than under high CO2 -low O2 air. Maximal ΦPSII showed no difference among either cultivars, gas mixtures or stress levels, the efficiency of excitation capture being highly resistant to drought. The ΦPSII evaluated during the transient yielded predictable values in respect of drought tolerance for durum wheat and barley cultivars, highlighting the key role of regulatory processes such as the Mehler peroxidase reaction and possibly also cyclic electron transport, in preventing overreduction under stress. The results clearly show that when Chl fluorescence analysis is used as a parameter in plant breeding, different experimental conditions should be used depending on the physiological mechanism that is bred or selected for. [ABSTRACT FROM AUTHOR]

Published
1998
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25. Multifunctional Core@Satellite Magnetic Particles for Magnetoresistive Biosensors

Author

Raffaele Campanile, Adriano Acunzo, Emanuela Scardapane, Antonio Minopoli, Veronica C. Martins, Rocco Di Girolamo, Susana Cardoso, Raffaele Velotta, Bartolomeo Della Ventura, Vincenzo Iannotti, Campanile, R., Acunzo, A., Scardapane, E., Minopoli, A., Martins, V. C., Di Girolamo, R., Cardoso, S., Velotta, R., Della Ventura, B., and Iannotti, V.

Subjects
General Chemical Engineering, General Chemistry
Abstract

Magnetoresistive (MR) biosensors combine distinctive features such as small size, low cost, good sensitivity, and propensity to be arrayed to perform multiplexed analysis. Magnetic nanoparticles (MNPs) are the ideal target for this platform, especially if modified not only to overcome their intrinsic tendency to aggregate and lack of stability but also to realize an interacting surface suitable for biofunctionalization without strongly losing their magnetic response. Here, we describe an MR biosensor in which commercial MNP clusters were coated with gold nanoparticles (AuNPs) and used to detect human IgG in water using an MR biochip that comprises six sensing regions, each one containing five U-shaped spin valve sensors. The isolated AuNPs (satellites) were stuck onto an aggregate of individual iron oxide crystals (core) so that the resulting core@satellite magnetic particles (CSMPs) could be functionalized by the photochemical immobilization technique an easy procedure that leads to oriented antibodies immobilized upright onto gold. The morphological, optical, hydrodynamic, magnetic, and surface charge properties of CSMPs were compared with those exhibited by the commercial MNP clusters showing that the proposed coating procedure endows the MNP clusters with stability and ductility without being detrimental to magnetic properties. Eventually, the high-performance MR biosensor allowed us to detect human IgG in water with a detection limit of 13 pM (2 ng mL-1). Given its portability, the biosensor described in this paper lends itself to a point-of-care device; moreover, the features of the MR biochip also make it suitable for multiplexed analysis.

Published
2022
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26. Polymer physics reveals a combinatorial code linking 3D chromatin architecture to 1D chromatin states

Author

Andrea Esposito, Simona Bianco, Andrea M. Chiariello, Alex Abraham, Luca Fiorillo, Mattia Conte, Raffaele Campanile, Mario Nicodemi, Esposito, A., Bianco, S., Chiariello, A. M., Abraham, A., Fiorillo, L., Conte, M., Campanile, R., and Nicodemi, M.

Subjects
Mammals, Genome, Animal, Polymers, Physics, Chromosome, biophysic, epigenomic, General Biochemistry, Genetics and Molecular Biology, Mammal, Chromatin, Chromosomes, machine learning, chromatin architecture, Cardiovascular and Metabolic Diseases, 3D genome organization, computer simulation, Physic, Animals, CP: Molecular biology, polymer-physic
Abstract

The mammalian genome has a complex, functional 3D organization. However, it remains largely unknown how DNA contacts are orchestrated by chromatin organizers. Here, we infer from only Hi-C the cell-type-specific arrangement of DNA binding sites sufficient to recapitulate, through polymer physics, contact patterns genome wide. Our model is validated by its predictions in a set of duplications at Sox9 against available independent data. The binding site types fall in classes that well match chromatin states from segmentation studies, yet they have an overlapping, combinatorial organization along chromosomes necessary to accurately explain contact specificity. The chromatin signatures of the binding site types return a code linking chromatin states to 3D architecture. The code is validated by extensive de novo predictions of Hi-C maps in an independent set of chromosomes. Overall, our results shed light on how 3D information is encrypted in 1D chromatin via the specific combinatorial arrangement of binding sites.

Published
2022

27. Efficient computational implementation of polymer physics models to explore chromatin structure

Author

Raffaele Campanile, Alfonso Corrado, Luca Fiorillo, Andrea M. Chiariello, Andrea Esposito, Mattia Conte, Maria Gabriella Chiariello, Carlo Annunziatella, Simona Bianco, Conte, M., Esposito, A., Fiorillo, L., Campanile, R., Annunziatella, C., Corrado, A., Chiariello, M. G., Bianco, S., and Chiariello, A. M.

Subjects
Physics, Molecular dynamics, Molecular dynamic, Sprite (lightning), Computer Networks and Communications, Spatial structure, Polymer physics, Biological system, polymer physics, Software, Chromatin, chromatin organization
Abstract

The development of novel experimental technologies able to map genome-wide chromatin contacts, as Hi-C, GAM or SPRITE, allowed to derive detailed information about the spatial structure of chromosomes in the cell nucleus. They revealed that the genome has a complex spatial organisation, which is highly connected with its activity. In the last years, such an abundance of experimental data prompted the development of quantitative models based on Polymer Physics to describe the chromatin architecture, clarifying many aspects about the molecular mechanisms underlying genome folding. Efficient algorithms are thus fundamental to perform massive numerical simulations for testing the accuracy of these models and provide a good description for small genomic regions or for whole chromosomes. Here, we consider the performances of Molecular Dynamics (MD) implementation of commonly used polymer physics models. Such models can be combined with Machine Learning approaches informed with experimental data to produce more accurate descriptions of real genomic regions. However, the execution times increase as a power-law with the size of the input data, which ultimately reflects the complexity of the investigated system. The best strategy is therefore a convenient trade-off between the accuracy in the description and the availability of computational resources. The combination of innovative experimental data and polymer physics theories allow to reconstruct the 3D genome structure. This is achieved by the use of machine learning approaches and massive parallel computing. Efficient algorithms and computational resources are then fundamental to produce models of increasingly high accuracy.

Published
2022

28. Magnetic Micromixing for Highly Sensitive Detection of Glyphosate in Tap Water by Colorimetric Immunosensor

Author

Raffaele Campanile, Valerio Cosimo Elia, Antonio Minopoli, Zaheer Ud Din Babar, Rocco di Girolamo, Antonio Morone, Nikola Sakač, Raffaele Velotta, Bartolomeo Della Ventura, Vincenzo Iannotti, Campanile, R., Elia, V. C., Minopoli, A., Ud Din Babar, Z., di Girolamo, R., Morone, A., Sakac, N., Velotta, R., Della Ventura, B., and Iannotti, V.

Subjects
Immunoassay, History, Glyphosate, Polymers and Plastics, Magnetic Phenomena, Magnetic nanoparticle, Water, Biosensing Techniques, Colorimetric immunosensor, Industrial and Manufacturing Engineering, Analytical Chemistry, Photochemical immobilization technique, Micromixing, Gold, Business and International Management, Core-shell nanoparticle
Abstract

Glyphosate is the most widely used herbicide in the world and, in view of its toxicity, there is a quest for easy-to-use, but reliable methods to detect it in water. To address this issue, we realized a simple, rapid, and highly sensitive immunosensor based on gold coated magnetic nanoparticles (MNPs@Au) to detect glyphosate in tap water. Not only the gold shell provided a sensitive optical transduction of the biological signal – through the shift of the local surface plasmon resonance (LSPR) entailed by the nanoparticle aggregation –, but it also allowed us to use an effective photochemical immobilization technique to tether oriented antibodies straight on the nanoparticles surface. While such a feature led to aggregates in which the nanoparticles were at close proximity each other, the magnetic properties of the core offered us an efficient tool to steer the nanoparticles by a rotating magnetic field. As a result, the nanoparticle aggregation in presence of the target could take place at higher rate (enhanced diffusion) with significant improvement in sensitivity. As a matter of fact, the combination of plasmonic and magnetic properties within the same nanoparticles allowed us to realize a colorimetric biosensor with a limit of detection (LOD) of 20 ng∙L−1.

Published
2022
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29. Gold Coated Nanoparticles Functionalized by Photochemical Immobilization Technique for Immunosensing

Author

Raffaele Campanile, Raffaele Velotta, A. Forente, Martina Cimafonte, Emanuela Scardapane, Antonio Minopoli, B. Della Ventura, V. Elia, Chiara Schiattarella, Della Ventura, B., Campanile, R., Cimafonte, M., Elia, V., Forente, A., Minopoli, A., Scardapane, E., Schiattarella, C., and Velotta, R.

Subjects
Localised surface plasmon resonance, Materials science, Colorimetric, 010401 analytical chemistry, Nanoparticle, 02 engineering and technology, 021001 nanoscience & nanotechnology, Photochemistry, 01 natural sciences, 0104 chemical sciences, Photochemical immobilization technique, Colloidal gold, Optical transducers, Magnetic nanoparticles, Surface modification, Gold surface, Core-shell nanoparticle, 0210 nano-technology, Biosensor, Plasmon
Abstract

Gold nanoparticles (AuNP)s play a fundamental role in biosensing in view of their various applications, each of them exploiting one or more properties of such a system. In any case, to make AuNPs effective as biosensing tools, it is necessary to provide them with high specificity by means of processes able to achieve an efficient gold surface functionalization with antibodies. This issue can be easily addressed by the so-called Photochemical Immobilization Technique (PIT). Herein, we report PIT-functionalized AuNPs applied as ballast for improving the sensitivity of quartz-crystal microbalances, but also as optical transducers (colorimetric biosensor) in which the key role is played by the plasmonic properties of nanostructured gold. Eventually, the role of gold is highlighted in its combination with magnetic nanoparticles, a physical system in which the magnetic behaviour of the core is joined to the optical properties of the surface.

Published
2021

30. Randomly positioned gold nanoparticles as fluorescence enhancers in apta-immunosensor for malaria test

Author

Raffaele Campanile, Andreas Offenhäusser, Antonio Minopoli, Julian A. Tanner, Dirk Mayer, Bartolomeo Della Ventura, Raffaele Velotta, Minopoli, A., Della Ventura, B., Campanile, R., Tanner, J. A., Offenhausser, A., Mayer, D., and Velotta, R.

Subjects
Nanoplasmonics, Analyte, Aptamer, Plasmodium falciparum, Protozoan Proteins, Nanochemistry, Nanoparticle, Metal Nanoparticles, 02 engineering and technology, Biosensing Techniques, 010402 general chemistry, 01 natural sciences, Analytical Chemistry, Nanoplasmonic, Antibody-aptamer biosensor, Limit of Detection, Humans, Fluorescent Dyes, Immunoassay, Original Paper, Chromatography, L-Lactate Dehydrogenase, Chemistry, Substrate (chemistry), Plasmon-enhanced fluorescence, Aptamers, Nucleotide, 021001 nanoscience & nanotechnology, Fluorescence, Gold nanoparticle array, 0104 chemical sciences, Malaria, Photochemical immobilization technique, Malaria marker, Colloidal gold, ddc:540, Gold, 0210 nano-technology, Biosensor, Antibodies, Immobilized
Abstract

A plasmon-enhanced fluorescence-based antibody-aptamer biosensor — consisting of gold nanoparticles randomly immobilized onto a glass substrate via electrostatic self-assembly — is described for specific detection of proteins in whole blood. Analyte recognition is realized through a sandwich scheme with a capture bioreceptor layer of antibodies — covalently immobilized onto the gold nanoparticle surface in upright orientation and close-packed configuration by photochemical immobilization technique (PIT) — and a top bioreceptor layer of fluorescently labelled aptamers. Such a sandwich configuration warrants not only extremely high specificity, but also an ideal fluorophore-nanostructure distance (approximately 10–15 nm) for achieving strong fluorescence amplification. For a specific application, we tested the biosensor performance in a case study for the detection of malaria-related marker Plasmodium falciparum lactate dehydrogenase (PfLDH). The proposed biosensor can specifically detect PfLDH in spiked whole blood down to 10 pM (0.3 ng/mL) without any sample pretreatment. The combination of simple and scalable fabrication, potentially high-throughput analysis, and excellent sensing performance provides a new approach to biosensing with significant advantages compared to conventional fluorescence immunoassays. Graphical abstract

Published
2020
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31. Colorimetric Test for Fast Detection of SARS-CoV-2 in Nasal and Throat Swabs

Author

Antonio Minopoli, Raffaele Velotta, Daniela Terracciano, Raffaele Campanile, Michele Cennamo, Sergio Bolletti Censi, Bartolomeo Della Ventura, Giuseppe Portella, DELLA VENTURA, Bartolomeo, Cennamo, M., Minopoli, A., Campanile, R., Censi Bolletti, S., Terracciano, D., Portella, G., and Velotta, R.

Subjects
Letter, viruses, Threshold Limit Value, Metal Nanoparticles, Nanoparticle, colorimetric biosensor, Biosensing Techniques, 02 engineering and technology, medicine.disease_cause, Polymerase Chain Reaction, 01 natural sciences, Viral Envelope Proteins, antibody, Threshold Limit Values, Membrane Protein, point-of-care device, Instrumentation, Coronavirus, Fluid Flow and Transfer Processes, Chemistry, 021001 nanoscience & nanotechnology, medicine.anatomical_structure, photochemical immobilization technique, Colloidal gold, Spike Glycoprotein, Coronavirus, colorimetric biosensors, Colorimetry, Coronavirus Infections, 0210 nano-technology, Viral load, Human, Photochemistry, Pneumonia, Viral, Bioengineering, Context (language use), Specimen Handling, Biosensing Technique, Metal Nanoparticle, Betacoronavirus, Throat, medicine, Humans, Pandemics, Detection limit, Betacoronaviru, Pandemic, Coronavirus Infection, SARS-CoV-2, Process Chemistry and Technology, 010401 analytical chemistry, COVID-19, Membrane Proteins, Gold standard (test), Virology, 0104 chemical sciences, Nasal Mucosa, gold nanoparticles, Pharynx, Gold, Biosensor, gold nanoparticle, Spike Glycoprotein, Coronaviru
Abstract

Mass testing is fundamental to face the pandemic caused by the coronavirus SARS-CoV-2 discovered at the end of 2019. To this aim, it is necessary to establish reliable, fast and cheap tools to detect viral particles in biological material so to identify the people capable to spread the infection. We demonstrate that a colorimetric biosensor based on gold nanoparticle (AuNP) interaction induced by SARS-CoV-2 lends itself as an outstanding tool for detecting viral particles in nasal and throat swabs. The extinction spectrum of a colloidal solution of multiple viral-target gold nanoparticles – AuNPs functionalized with antibodies targeting three surface proteins of SARS-CoV-2 (spike, envelope and membrane) – is redshifted in few minutes when mixed to a solution containing the viral particle. The optical density of the mixed solution measured at 560 nm was compared to the threshold cycle (Ct) of a Real Time-PCR (gold standard for detecting the presence of viruses) finding that the colorimetric method is able to detect very low viral load with a detection limit approaching that of RT-PCR. Since the method is sensitive to the infecting viral particle rather than to its RNA, the achievements reported here open new perspective not only in the context of the current and possible future pandemics, but also in microbiology as the biosensor proves itself to be a powerful though simple tool for measuring the viral particle concentration.

Published
2020
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32. LSPR-based colorimetric immunosensor for rapid and sensitive 17β-estradiol detection in tap water

Author

Raffaele Campanile, Antonio Minopoli, Andreas Offenhäusser, Bohdan Lenyk, Raffaele Velotta, Nikola Sakač, Dirk Mayer, Bartolomeo Della Ventura, Minopoli, A., Sakac, N., Lenyk, B., Campanile, R., Mayer, D., Offenhausser, A., Velotta, R., and Della Ventura, B.

Subjects
Materials science, Gold nanoparticle, Antibodie, Nanoparticle, 02 engineering and technology, Colorimetric immunosensor, 010402 general chemistry, 01 natural sciences, Dynamic light scattering, Tap water, Scanning transmission electron microscopy, Materials Chemistry, Electrical and Electronic Engineering, Surface plasmon resonance, Instrumentation, Detection limit, Chromatography, Metals and Alloys, 17β-estradiol, Localized surface plasmon resonance, 021001 nanoscience & nanotechnology, Condensed Matter Physics, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, Highly sensitive, Photochemical immobilization technique, Colloidal gold, Gold nanoparticlesLocalized surface plasmon resonanceColorimetric immunosensor17β-estradiolPhotochemical immobilization techniqueAntibodies, ddc:620, 0210 nano-technology
Abstract

We propose a highly sensitive immunosensor based on the Localized Surface Plasmon Resonance (LSPR) for 17β-estradiol (E2) quantification in water. E2 molecules are recognized by polyclonal antibodies immobilized onto gold nanoparticles (AuNPs) and act as linkers that cause nanoparticles aggregation. This leads to the change in the optical properties of the solution visible even by naked eyes. The aggregates were characterized by Dynamic Light Scattering (DLS) and Scanning Transmission Electron Microscopy (STEM) that provided an accurate assessment of the inter-particle distance. The finite-difference time-domain (FDTD) method applied to a Mie problem like workspace allowed us to describe the optical behaviour of the AuNP aggregates with excellent agreement between the experimental and numerical results. The limit of detection (LOD), without any preconcentration step, is 3 pg/mL (11 pM), whereas the detection range extends over five decades up to 105 pg/mL. The proposed E2 immunosensor was tested in tap water, where no significant cross-reaction signal was detected by similar molecules (testosterone, progesterone, estrone and estriol). The device described here represents a significant improvement of low E2 levels determination in terms of affordability, time and measuring simplicity, making it suitable for environmental applications.

Published
2020
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33. Understanding chromatin structure: Efficient computational implementation of polymer physics models

Author

Raffaele Campanile, Mattia Conte, Luca Fiorillo, Carlo Annunziatella, Andrea Esposito, Simona Bianco, Andrea M. Chiariello, Bianco, S., Annunziatella, C., Esposito, A., Fiorillo, L., Conte, M., Campanile, R., and Chiariello, A. M.

Subjects
0301 basic medicine, chemistry.chemical_classification, Sequence, Molecular dynamic, Theoretical computer science, Computer science, Folding (DSP implementation), Polymer, Genome, Chromatin, 03 medical and health sciences, Cell nucleus, Molecular dynamics, 030104 developmental biology, 0302 clinical medicine, medicine.anatomical_structure, chemistry, medicine, Polymer physics, Focus (optics), 030217 neurology & neurosurgery
Abstract

In recent years the development of novel technologies, as Hi-C or GAM, allowed to investigate the spatial structure of chromatin in the cell nucleus with a constantly increasing level of accuracy. Polymer physics models have been developed and improved to better interpret the wealth of complex information coming from the experimental data, providing highly accurate understandings on chromatin architecture and on the mechanisms regulating genome folding. To investigate the capability of the models to explain the experiments and to test their agreement with the data, massive parallel simulations are needed and efficient algorithms are fundamental. In this work, we consider general computational Molecular Dynamics (MD) techniques commonly used to implement such models, with a special focus on the Strings & Binders Switch polymer model. By combining this model with machine learning computational approaches, it is possible to give an accurate description of real genomic loci. In addition, it is also possible to make predictions about the impact of structural variants of the genomic sequence, which are known to be linked to severe congenital diseases.

Published
2019

34. Neo-adjuvant chemotherapy does not increase the rate of complete resection and does not significantly reduce the morbidity of Visceral-Peritoneal Debulking (VPD) in patients with stage IIIC-IV ovarian cancer

Author

Giorgio Fachechi, Hooman Soleymani Majd, Kumar Gubbala, Federico Ferrari, Igor Martinek, Raffaella Giannice, Sarah Tardino, Riccardo Garruto Campanile, Stefano Cianci, Roberto Tozzi, Tozzi, R., Giannice, R., Cianci, S., Tardino, S., Campanile, R. G., Gubbala, K., Fachechi, G., Ferrari, F., Martinek, I., and Soleymani Majd, H.

Subjects
Complete resection, Adult, medicine.medical_specialty, medicine.medical_treatment, Carcinoma, Ovarian Epithelial, Young Adult, Ovarian cancer, Statistical significance, Ovarian Epithelial, Neoplasms, 80 and over, Medicine, Chemotherapy, Humans, Stage IIIC, Neoplasms, Glandular and Epithelial, Stage (cooking), Laparoscopy, Adjuvant, Aged, Neoplasm Staging, Aged, 80 and over, Porta hepatis, Ovarian Neoplasms, medicine.diagnostic_test, business.industry, Ovarian Neoplasm, Carcinoma, Obstetrics and Gynecology, Glandular and Epithelial, Middle Aged, Morbidity, medicine.disease, Debulking, Neoadjuvant Therapy, Surgery, medicine.anatomical_structure, Oncology, Chemotherapy, Adjuvant, Female, business, Human
Abstract

Objective To measure the efficacy and the safety of Visceral–Peritoneal Debulking (VPD) in patients with stage IIIC–IV ovarian cancer and to compare the outcomes before and after chemotherapy. Methods Between 2008 and 2013, 200 consecutive patients were offered VPD for stage IIIC/IV ovarian cancer. Exclusion criteria were: metastases in the lungs or 3 liver segments at CT review and/or disease on small bowel serosa or encasing the porta hepatis at explorative laparoscopy. The endpoints were efficacy (rate of complete resection, CR) and safety (morbidity and mortality). The results were compared between patients in group 1 (upfront surgery) and group 2 (during or after chemotherapy). Results Ninety-eight patients were in group 1 and 102 in group 2. Twenty out of 200 patients (10%) did not have VPD, 180 out of 200 patients (90%) had VPD and CR: 90.8% in group 1, 89.8% in group 2. The mortality (1%) and intra-operative complication rate (3.3%) were similar. Post-operative complications rate was 34.8% in group 1 vs. 30.7% in group 2 (P=0.669). The difference in grade III (15.7% vs. 5.5%, P=0.053) and grade IIIb complications (13.4% vs. 4.4%, P=0.062) approached statistical significance. All other outcomes were not significantly different in the 2 groups. Conclusion VPD achieved CR in 90% of the patients. Neo-adjuvant chemotherapy did not increase the rate of CR and did not significantly decrease the morbidity or the complexity of the surgery.

Published
2015

35. Pediatric portal vein thrombosis: more on thrombophilic risk factors

Author

S. Maddaluno, Carmelo Piscopo, Pietro Vajro, Giuseppe Castaldo, Riccardo Campanile, M. Caropreso, Vajro, P, Campanile, R, Caropreso, M, Piscopo, Carmelo, Maddaluno, S, and Castaldo, Giuseppe

Subjects
Male, Venous Thrombosis, medicine.medical_specialty, business.industry, Portal Vein, Gastroenterology, Portal vein, medicine.disease, Thrombophilia, Portal vein thrombosis, Venous thrombosis, Internal medicine, Pediatrics, Perinatology and Child Health, Mutation (genetic algorithm), Mutation, medicine, Humans, Female, business
Published
2013

36. Genetic prothrombotic risk factors in children with extrahepatic portal vein obstruction

Author

S. Maddaluno, M. Caropreso, Carmelo Piscopo, Giuseppe Castaldo, Claudio Veropalumbo, Pietro Vajro, Riccardo Campanile, Caropreso, Maria, Campanile, R, Maddaluno, Sergio, Veropalumbo, Claudio, Piscopo, Carmelo, Castaldo, Giuseppe, and Vajro, Pietro

Subjects
medicine.medical_specialty, Text mining, business.industry, Pediatrics, Perinatology and Child Health, Gastroenterology, medicine, Radiology, Portal vein obstruction, business, medicine.disease, Thrombosis
Published
2010

37. Production of AFM wedged cantilevers for stress-relaxation experiments: Uniaxial loading of soft, spherical cells.

Author

Campanile R, Helenius J, Scielzo C, Scarfò L, Salerno D, Bossi M, Falappi M, Saponara A, Müller DJ, Mantegazza F, and Cassina V

Abstract

The fabrication of wedge-shaped cantilevers for Atomic Force Microscopy (AFM) remains a critical yet challenging task, particularly when precision and efficiency are required. In this study, we present a streamlined protocol for producing these wedges using NOA63 UV-curing polymer, which simplifies the process and eliminates the need for dedicated equipment. Our method reduces preparation time while maintaining the mechanical properties of the cantilevers, in line with the manufacturer's specifications. We demonstrate the effectiveness of our wedged cantilevers in stress-relaxation experiments performed by means of AFM and confocal microscopy on primary Chronic Lymphocytic Leukemia cells and the MEC1 cell line. These experiments highlight the effectiveness of using modified cantilevers to consistently apply precise uniaxial loading to soft, spherical cells. This technique offers a marked improvement in fabrication speed and operational ease compared to traditional methods, without compromising the accuracy or performance of the measurements. This protocol is not only time-saving, but also adaptable for use in a wide range of biological applications, making it a valuable tool for AFM-based research in cellular mechanics., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Inc. All rights reserved.)

Published
2025
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38. The Nanomechanical Properties of CLL Cells Are Linked to the Actin Cytoskeleton and Are a Potential Target of BTK Inhibitors.

Author

Sampietro M, Cassina V, Salerno D, Barbaglio F, Buglione E, Marrano CA, Campanile R, Scarfò L, Biedenweg D, Fregin B, Zamai M, Díaz Torres A, Labrador Cantarero V, Ghia P, Otto O, Mantegazza F, Caiolfa VR, and Scielzo C

Abstract

Chronic lymphocytic leukemia (CLL) is an incurable disease characterized by an intense trafficking of the leukemic cells between the peripheral blood and lymphoid tissues. It is known that the ability of lymphocytes to recirculate strongly depends on their capability to rapidly rearrange their cytoskeleton and adapt to external cues; however, little is known about the differences occurring between CLL and healthy B cells during these processes. To investigate this point, we applied a single-cell optical (super resolution microscopy) and nanomechanical approaches (atomic force microscopy, real-time deformability cytometry) to both CLL and healthy B lymphocytes and compared their behavior. We demonstrated that CLL cells have a specific actomyosin complex organization and altered mechanical properties in comparison to their healthy counterpart. To evaluate the clinical relevance of our findings, we treated the cells in vitro with the Bruton's tyrosine kinase inhibitors and we found for the first time that the drug restores the CLL cells mechanical properties to a healthy phenotype and activates the actomyosin complex. We further validated these results in vivo on CLL cells isolated from patients undergoing ibrutinib treatment. Our results suggest that CLL cells' mechanical properties are linked to their actin cytoskeleton organization and might be involved in novel mechanisms of drug resistance, thus becoming a new potential therapeutic target aiming at the normalization of the mechanical fingerprints of the leukemic cells., Competing Interests: OO is the cofounder of Zellmechanik Dresden commercialising real-time deformability cytometry. PG is a HemaSphere editor. All the other authors have no conflicts of interest to disclose., (Copyright © 2023 the Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the European Hematology Association.)

Published
2023
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39. Magnetic micromixing for highly sensitive detection of glyphosate in tap water by colorimetric immunosensor.

Author

Campanile R, Elia VC, Minopoli A, Ud Din Babar Z, di Girolamo R, Morone A, Sakač N, Velotta R, Della Ventura B, and Iannotti V

Subjects
Immunoassay, Water, Gold, Magnetic Phenomena, Biosensing Techniques
Abstract

Glyphosate is the most widely used herbicide in the world and, in view of its toxicity, there is a quest for easy-to-use, but reliable methods to detect it in water. To address this issue, we realized a simple, rapid, and highly sensitive immunosensor based on gold coated magnetic nanoparticles (MNPs@Au) to detect glyphosate in tap water. Not only the gold shell provided a sensitive optical transduction of the biological signal - through the shift of the local surface plasmon resonance (LSPR) entailed by the nanoparticle aggregation -, but it also allowed us to use an effective photochemical immobilization technique to tether oriented antibodies straight on the nanoparticles surface. While such a feature led to aggregates in which the nanoparticles were at close proximity each other, the magnetic properties of the core offered us an efficient tool to steer the nanoparticles by a rotating magnetic field. As a result, the nanoparticle aggregation in presence of the target could take place at higher rate (enhanced diffusion) with significant improvement in sensitivity. As a matter of fact, the combination of plasmonic and magnetic properties within the same nanoparticles allowed us to realize a colorimetric biosensor with a limit of detection (LOD) of 20 ng∙L -1 ., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Published
2023
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40. Exploring the Link between ADHD and Obesity: A Focus on Temperament.

Author

Porfirio MC, Campanile R, Masi G, Purper-Ouakil D, Giovinazzo S, Ascenzi A, Troisi A, and Mazzone L

Abstract

Multiple studies support the relationship between ADHD and overweight/obesity in youth. Different mechanisms may be involved, such as temperamental and psychopathological factors. The aim of this study was to test the hypothesis that specific temperamental and psychopathological dimensions could mediate the relationship between ADHD and obesity. The sample included 100 children and adolescents (78 males and 22 females; age range 6 to 18 years; mean age 9.90 ± 2.5 years). The assessment procedure included Conners' Parent Rating Scale-Long (CPRS-R:L) as the inclusion criterion for ADHD diagnosis, the Child Behavior Checklist (CBCL), a dimensional measure for psychopathology, and the Junior Temperament and Character Inventory, which describes four temperamental dimensions: novelty seeking (NS), harm avoidance (HA), reward dependence (RD), and persistence (P). While in the whole ADHD sample, the highest scores were found in NS and the lowest in P, ADHD with overweight/obesity, compared to ADHD with normal weight, showed higher HA and RD, lower NS, and higher CBCL Internalizing scores. These findings suggest that ADHD youth with overweight/obesity present specific temperamental and psychopathological features compared to those without overweight/obesity. If confirmed in larger samples, using a control group without ADHD, these temperamental and psychological features may be helpful for an earlier recognition of ADHD patients at higher risk for obesity, and may represent possible targets for temperament-based preventive interventions and tailored treatment programs. These features should be included in the routine assessment of children and adolescents with ADHD and/or are overweight/obese.

Published
2022
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41. Polymer physics reveals a combinatorial code linking 3D chromatin architecture to 1D chromatin states.

Author

Esposito A, Bianco S, Chiariello AM, Abraham A, Fiorillo L, Conte M, Campanile R, and Nicodemi M

Subjects
Animals, Chromosomes, Genome, Mammals genetics, Physics, Chromatin, Polymers
Abstract

The mammalian genome has a complex, functional 3D organization. However, it remains largely unknown how DNA contacts are orchestrated by chromatin organizers. Here, we infer from only Hi-C the cell-type-specific arrangement of DNA binding sites sufficient to recapitulate, through polymer physics, contact patterns genome wide. Our model is validated by its predictions in a set of duplications at Sox9 against available independent data. The binding site types fall in classes that well match chromatin states from segmentation studies, yet they have an overlapping, combinatorial organization along chromosomes necessary to accurately explain contact specificity. The chromatin signatures of the binding site types return a code linking chromatin states to 3D architecture. The code is validated by extensive de novo predictions of Hi-C maps in an independent set of chromosomes. Overall, our results shed light on how 3D information is encrypted in 1D chromatin via the specific combinatorial arrangement of binding sites., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022. Published by Elsevier Inc.)

Published
2022
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42. Impact of the COVID-19 Pandemic on Child and Adolescent Psychiatric Emergencies.

Author

Davico C, Marcotulli D, Lux C, Calderoni D, Cammisa L, Bondone C, Rosa-Brusin M, Secci I, Porro M, Campanile R, Bosia C, Di Santo F, Terrinoni A, Ricci F, Amianto F, Urbino A, Ferrara M, and Vitiello B

Subjects
Adolescent, Age Factors, Child, Communicable Disease Control methods, Education, Distance, Female, Humans, Italy epidemiology, Male, Mental Health statistics & numerical data, Organizational Innovation, SARS-CoV-2, Ambulatory Care statistics & numerical data, COVID-19 epidemiology, COVID-19 prevention & control, COVID-19 psychology, Emergencies epidemiology, Emergency Services, Psychiatric organization & administration, Emergency Services, Psychiatric statistics & numerical data, Hospitalization statistics & numerical data, Mental Disorders epidemiology, Mental Disorders psychology, Mental Disorders therapy, Physical Distancing
Abstract

Objective: By forcing closure of schools, curtailing outpatient services, and imposing strict social distancing, the COVID-19 pandemic has abruptly affected the daily life of millions worldwide, with still unclear consequences for mental health. This study aimed to evaluate if and how child and adolescent psychiatric visits to hospital emergency departments (EDs) changed during the pandemic lockdown, which started in Italy on February 24, 2020., Methods: We examined all ED visits by patients under 18 years of age in the 7 weeks prior to February 24, 2020, and in the subsequent 8 weeks of COVID-19 lockdown at two urban university hospitals, in Turin and Rome, Italy. ED visits during the corresponding periods of 2019 served as a comparison using Poisson regression modeling. The clinician's decision to hospitalize or discharge home the patient after the ED visit was examined as an index of clinical severity., Results: During the COVID-19 lockdown, there was a 72.0% decrease in the number of all pediatric ED visits (3,395) compared with the corresponding period in 2019 (12,128), with a 46.2% decrease in psychiatric visits (50 vs 93). The mean age of psychiatric patients was higher in the COVID-19 period (15.7 vs 14.1 years). No significant changes were found in hospitalization rate or in the prevalence distribution of the primary reason for the psychiatric ED visit (suicidality, anxiety/mood disorders, agitation)., Conclusions: In the first 8 weeks of the COVID-19-induced social lockdown, the number of child and adolescent psychiatric ED visits significantly decreased, with an increase in patient age. This decrease does not appear to be explained by severity-driven self-selection and might be due to a reduction in psychiatric emergencies or to the implementation of alternative ways of managing acute psychopathology., (© Copyright 2021 Physicians Postgraduate Press, Inc.)

Published
2021
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43. Colorimetric Test for Fast Detection of SARS-CoV-2 in Nasal and Throat Swabs.

Author

Ventura BD, Cennamo M, Minopoli A, Campanile R, Censi SB, Terracciano D, Portella G, and Velotta R

Subjects
Biosensing Techniques, COVID-19, Gold, Humans, Membrane Proteins chemistry, Metal Nanoparticles, Pandemics, Photochemistry, Polymerase Chain Reaction, SARS-CoV-2, Specimen Handling, Spike Glycoprotein, Coronavirus chemistry, Threshold Limit Values, Viral Envelope Proteins chemistry, Betacoronavirus chemistry, Colorimetry methods, Coronavirus Infections diagnosis, Coronavirus Infections virology, Nasal Mucosa virology, Pharynx virology, Pneumonia, Viral diagnosis, Pneumonia, Viral virology
Abstract

Mass testing is fundamental to face the pandemic caused by the coronavirus SARS-CoV-2 discovered at the end of 2019. To this aim, it is necessary to establish reliable, fast, and cheap tools to detect viral particles in biological material so to identify the people capable of spreading the infection. We demonstrate that a colorimetric biosensor based on gold nanoparticle (AuNP) interaction induced by SARS-CoV-2 lends itself as an outstanding tool for detecting viral particles in nasal and throat swabs. The extinction spectrum of a colloidal solution of multiple viral-target gold nanoparticles-AuNPs functionalized with antibodies targeting three surface proteins of SARS-CoV-2 (spike, envelope, and membrane)-is red-shifted in few minutes when mixed with a solution containing the viral particle. The optical density of the mixed solution measured at 560 nm was compared to the threshold cycle ( C t ) of a real-time PCR (gold standard for detecting the presence of viruses) finding that the colorimetric method is able to detect very low viral load with a detection limit approaching that of the real-time PCR. Since the method is sensitive to the infecting viral particle rather than to its RNA, the achievements reported here open a new perspective not only in the context of the current and possible future pandemics, but also in microbiology, as the biosensor proves itself to be a powerful though simple tool for measuring the viral particle concentration.

Published
2020
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44. Core-Shell Magnetic Nanoparticles for Highly Sensitive Magnetoelastic Immunosensor.

Author

Campanile R, Scardapane E, Forente A, Granata C, Germano R, Di Girolamo R, Minopoli A, Velotta R, Della Ventura B, and Iannotti V

Abstract

A magnetoelastic (ME) biosensor for wireless detection of analytes in liquid is described. The ME biosensor was tested against human IgG in the range 0-20 μg∙mL -1 . The sensing elements, anti-human IgG produced in goat, were immobilized on the surface of the sensor by using a recently introduced photochemical immobilization technique (PIT), whereas a new amplification protocol exploiting gold coated magnetic nanoparticles (core-shell nanoparticles) is demonstrated to significantly enhance the sensitivity. The gold nanoflowers grown on the magnetic core allowed us to tether anti-human IgG to the nanoparticles to exploit the sandwich detection scheme. The experimental results show that the 6 mm × 1 mm × 30 μm ME biosensor with an amplification protocol that uses magnetic nanoparticles has a limit of detection (LOD) lower than 1 nM, works well in water, and has a rapid response time of few minutes. Therefore, the ME biosensor is very promising for real-time wireless detection of pathogens in liquids and for real life diagnostic purpose.

Published
2020
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45. The Repertoire of Serous Ovarian Cancer Non-genetic Heterogeneity Revealed by Single-Cell Sequencing of Normal Fallopian Tube Epithelial Cells.

Author

Hu Z, Artibani M, Alsaadi A, Wietek N, Morotti M, Shi T, Zhong Z, Santana Gonzalez L, El-Sahhar S, Carrami EM, Mallett G, Feng Y, Masuda K, Zheng Y, Chong K, Damato S, Dhar S, Campo L, Garruto Campanile R, Soleymani Majd H, Rai V, Maldonado-Perez D, Jones S, Cerundolo V, Sauka-Spengler T, Yau C, and Ahmed AA

Subjects
Cystadenocarcinoma, Serous genetics, Cystadenocarcinoma, Serous metabolism, Cystadenocarcinoma, Serous pathology, Epithelium metabolism, Epithelium pathology, Fallopian Tube Neoplasms genetics, Fallopian Tube Neoplasms pathology, Fallopian Tubes metabolism, Female, Gene Expression Regulation, Neoplastic genetics, Genetic Heterogeneity, Humans, Ovarian Neoplasms metabolism, Epithelial Cells pathology, Fallopian Tube Neoplasms metabolism, Fallopian Tubes pathology, Ovarian Neoplasms pathology
Abstract

The inter-differentiation between cell states promotes cancer cell survival under stress and fosters non-genetic heterogeneity (NGH). NGH is, therefore, a surrogate of tumor resilience but its quantification is confounded by genetic heterogeneity. Here we show that NGH in serous ovarian cancer (SOC) can be accurately measured when informed by the molecular signatures of the normal fallopian tube epithelium (FTE) cells, the cells of origin of SOC. Surveying the transcriptomes of ∼6,000 FTE cells, predominantly from non-ovarian cancer patients, identified 6 FTE subtypes. We used subtype signatures to deconvolute SOC expression data and found substantial intra-tumor NGH. Importantly, NGH-based stratification of ∼1,700 tumors robustly correlated with survival. Our findings lay the foundation for accurate prognostic and therapeutic stratification of SOC., Competing Interests: Declaration of Interests A.A.A., C.Y., and Z.H. filed a patent application for the use of the 52-gene panel for predicting the prognosis in ovarian cancer., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Published
2020
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46. Morbidity of multiple bowel resection compared to single bowel resection after debulking surgery for ovarian cancer.

Author

Tozzi R, Casarin J, Baysal A, Pinelli C, Matak L, Ghanbarzadeh N, Alazzam M, Garruto-Campanile R, Majd HS, Kilic Y, and Morotti M

Subjects
Aged, Carcinoma, Ovarian Epithelial drug therapy, Chemotherapy, Adjuvant, Cytoreduction Surgical Procedures adverse effects, Digestive System Surgical Procedures adverse effects, Female, Humans, Middle Aged, Ovarian Neoplasms drug therapy, Postoperative Complications etiology, Carcinoma, Ovarian Epithelial surgery, Cytoreduction Surgical Procedures methods, Digestive System Surgical Procedures methods, Intestines surgery, Ovarian Neoplasms surgery
Abstract

Objectives: To assess the impact of multiple bowel resections on postoperative outcomes in stage IIIC-IV ovarian cancer (OC)., Methods: From the Oxford OC database we retrieved consecutive patients who underwent bowel resection between January 2009 and November 2017. Patients were divided into two groups: single bowel resection (SBR) and MBR (≥2 bowel resections). The following outcomes were compared between the two groups: 30-day related and not related morbidity to bowel surgery, bowel diversion rate and time to start/restart adjuvant chemotherapy., Results: Thirty-five patients were in the MBR and 146 in the SBR group. The 30-day overall surgical-related complication and bowel specific complications rate was higher in MBR group than SBR group (54.3% vs. 23.9%, p < 0.001) and (25.7% vs. 10.5%, p = 0.035), respectively. The rate of bowel diversion was 97.7% in MBR vs. 26.7% in the SBR group (p = 0.021). Trend analysis showed a significant reduction in the rate of MBR after the introduction of NACT (p- for trend <0.001)., Conclusions: Our data show that MBR during OC surgery is associated with a higher rate of overall and bowel specific complication compared to SBR. The introduction of NACT is associated with a reduced rate of MBR., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Published
2019
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47. Modeling Single-Molecule Conformations of the HoxD Region in Mouse Embryonic Stem and Cortical Neuronal Cells.

Author

Bianco S, Annunziatella C, Andrey G, Chiariello AM, Esposito A, Fiorillo L, Prisco A, Conte M, Campanile R, and Nicodemi M

Subjects
Animals, Mice, Molecular Conformation, Embryonic Stem Cells metabolism, Neurons metabolism
Abstract

Complex architectural rearrangements are associated to the control of the HoxD genes in different cell types; yet, how they are implemented in single cells remains unknown. By use of polymer models, we dissect the locus 3D structure at the single DNA molecule level in mouse embryonic stem and cortical neuronal cells, as the HoxD cluster changes from a poised to a silent state. Our model describes published Hi-C, 3-way 4C, and FISH data with high accuracy and is validated against independent 4C data on the Nsi-SB 0.5-Mb duplication and on triple contacts. It reveals the mode of action of compartmentalization on the regulation of the HoxD genes that have gene- and cell-type-specific multi-way interactions with their regulatory elements and high cell-to-cell variability. It shows that TADs and higher-order 3D structures, such as metaTADs, associate with distinct combinations of epigenetic factors, including but not limited to CCCTC-binding factor (CTCF) and histone marks., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Published
2019
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48. Concomitant Laparoscopic and Thoracoscopic Resection of Recurrent High-Grade Ovarian Cancer.

Author

Tozzi R, Casarin J, Belcher E, Garruto Campanile R, Ismail L, Soleymani Majd H, Morotti M, and Desgro M

Subjects
Antineoplastic Combined Chemotherapy Protocols therapeutic use, Blood Loss, Surgical, Carboplatin administration & dosage, Carcinoma, Endometrioid drug therapy, Chemotherapy, Adjuvant, Female, Humans, Laparoscopy methods, Middle Aged, Neoplasm Recurrence, Local drug therapy, Operative Time, Ovarian Neoplasms drug therapy, Paclitaxel administration & dosage, Tomography, X-Ray Computed, Carcinoma, Endometrioid surgery, Neoplasm Recurrence, Local surgery, Ovarian Neoplasms surgery, Thoracoscopy methods
Abstract

Study Objective: To describe the first case of combined endoscopic management of a thoracic and abdominal recurrence of ovarian cancer., Design: An instructive video showing the combined thoracic and abdominal surgical procedure., Setting: Department of Gynecological Oncology, Churchill Hospital, Oxford University, UK., Patients: A 64-year-old woman undergoing endoscopic treatment for a third recurrence of ovarian cancer after a full surgical staging in 2007. The disease-free interval from the last recurrence was 31 months., Intervention: The operation was performed by a multidisciplinary team of thoracic and gynecologic oncologist surgeons. Surgery started with thoracoscopic resection of a right enlarged paracardiac lymph node of 24 mm and a small wedge of the right lung, which was attached to the lymph node. At laparoscopy, 2 nodules of 3 and 5 mm were excised from the mesosigmoid and 1 nodule of 20 mm was resected from the right hemidiaphragm., Measurements and Main Results: The total operative time was 251 minutes, and no intraoperative complication occurred. No conversion to open surgery was necessary. The estimated blood loss was 50 mL. There was no visible residual disease at the end of the surgery. The patient was discharged 4 days after surgery. The final pathology report confirmed the presence of endometrioid adenocarcinoma in all specimens removed. Adjuvant chemotherapy with carboplatin/paclitaxel was started 2 weeks later. At the 60-day follow-up, no complications were recorded. A computed tomographic scan performed after 6 cycles of chemotherapy did not reveal any evidence of relapse., Conclusions: The combined endoscopic approach might be feasible in selected patients., (Copyright © 2018 American Association of Gynecologic Laparoscopists. Published by Elsevier Inc. All rights reserved.)

Published
2018
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49. Morbidity and reversal rate of ileostomy after bowel resection during Visceral-Peritoneal Debulking (VPD) in patients with stage IIIC-IV ovarian cancer.

Author

Tozzi R, Casarin J, Garruto-Campanile R, Majd HS, and Morotti M

Subjects
Colon, Sigmoid surgery, Female, Humans, Middle Aged, Morbidity, Neoplasm Staging, Ovarian Neoplasms pathology, Ileostomy adverse effects, Ileostomy methods, Ovarian Neoplasms surgery, Rectum surgery
Abstract

Objective: To investigate the morbidity of diverting loop ileostomy (DLI) performed during Visceral Peritoneal Debulking (VPD) for stage IIIC-IV ovarian cancer and to report the rate, timing, and morbidity of DLI reversal., Methods: We retrieved the data of all consecutive patients who underwent sigmoid-rectum resection (SRR) followed by DLI. Morbidity was defined as any surgical/medical complications clearly correlated to the DLI. The reversal rate of DLI was defined as the number of patients who had the continuity of the gastrointestinal tract restored in the study period. Finally, we recorded the timing and the morbidity of the reversal surgery. Factors associated with non-reversal of DLI were reported., Results: In the study period (01/2010-09/2016), complete data were available for 47 patients. Stoma-related complications occurred in 22 patients (46.8%). Eight patients (17.0%) were readmitted within 30days from surgery. Thirty-two patients (68.1%) had their stoma reversed. The primary cause of non-reversal was tumor recurrence/progression (7/15, 46.7%). Patient's age, length of hospitalization, complications after VPD were associated with non-reversal of DLI. The mean time from DLI formation to stoma reversal was 6months (±1.7). Post-reversal related complications occurred in 37.1% of the patients., Conclusions: In our series, 31.9% of the patients with FIGO stage IIIC-IV ovarian cancer who underwent SRR and DLI did not have stoma reversal. Overall they had approximately 45% risk of stoma-related morbidity and 37% risk of morbidity related to the stoma reversal. This information should be part of the consulting process when preparing for debulking surgery, particularly in patients who are likely to need a bowel resection., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Published
2018
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50. En-bloc resection of the pelvis (EnBRP) in patients with stage IIIC-IV ovarian cancer: A 10 steps standardised technique. Surgical and survival outcomes of primary vs. interval surgery.

Author

Tozzi R, Hardern K, Gubbala K, Garruto Campanile R, and Soleymani Majd H

Subjects
Female, Humans, Neoplasm Staging, Ovarian Neoplasms mortality, Ovarian Neoplasms pathology, Retrospective Studies, Survival Analysis, Treatment Outcome, Cytoreduction Surgical Procedures methods, Ovarian Neoplasms surgery, Pelvis surgery
Abstract

Objective: In this study we describe the technique of the En-bloc resection of the pelvis (EnBRP) in 10 standardised and reproducible steps, whereby all pelvic organs, except the bladder, are removed together with the peritoneum. In addition, we compare the surgical and survival outcomes of patients who underwent upfront vs. interval surgery., Methods: Retrospective analysis of patients with FIGO Stage IIIC-IV ovarian cancer treated with Visceral Peritoneal Debulking (VPD) who had EnBRP. The study population was divided into Group 1 (up-front VPD) and group 2 (VPD after neo-adjuvant chemotherapy). The aim was to assess the incidence of EnBRP. We also assessed rate of complete resection (CR), procedure-specific and overall morbidity, disease free and overall survival. Results were compared between group 1 and 2., Results: Overall 92 out of 200 patients (46%) needed an EnBRP during the VPD. Forty-eight patients were in Group 1 and 44 patients in Group 2. CR was achieved in all patients. No intra-operative procedure specific morbidity was recorded. Dehiscence of bowel anastomosis was the only procedure specific morbidity. Rate was 2%, with 1 episode recorded in each group. Both patients were managed and settled with formation of a bowel diversion. The overall morbidity rate was 33%, 35% in group 1 and 31% in group 2. The mortality rate was 1%. Median disease free survival was 20months, 25 in group 1 vs. 15 in group 2 (P=0.009)., Conclusions: EnBRP is a safe and effective technique to tackle the pelvic disease of patients with advanced ovarian cancer. The reduced blood loss, the high rate of clear margins and CR of the disease are accompanied by a low rate of surgical morbidity. These features are particularly suitable for patients who are due to start or re-start chemotherapy. The standardization of the technique will make it more reproducible and easier to be taught. In addition, it will facilitate comparison of results and the inclusion of this technique in the portfolio of procedures as part of debulking surgery., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Published
2017
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