Publications by authors named "Federico Boschi"

78 Publications

Photon emission and changes in fluorescent properties of bone after laser irradiation.

J Biophotonics 2021 Mar 11:e202000445. Epub 2021 Mar 11.

Department of Computer Science, University of Verona, Verona, Italy.

Laser scalpels used in medical surgery concentrate light energy, heating the tissues. Recently, we reported thermoluminescence emission from laser-treated soft tissues. Here we investigated the thermo-optical effects caused by a laser operating at 808 nm on animal bones (beef ribs) through luminescence and fluorescence imaging, thermal imaging and scanning electron microscopy. Laser-induced artificial lesions emitted luminescence peaking around 650 nm, with a half-life of almost 1 hour. As concerns fluorescence, 24 hours after laser treatment we observed an increase of the emission and a shift from 500 (untreated) to 580 nm (treated). Recrystallization observed by SEM indicates that the temperature in the artificial lesions is over 600°C. We can conclude that laser treatment induces specific luminescent and fluorescent emissions due to heating of the bone and modification of its components. Monitoring these emissions could help prevent tissue overheating and its potential damages during laser-assisted medical procedures.
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http://dx.doi.org/10.1002/jbio.202000445DOI Listing
March 2021

High activity and low toxicity of a novel CD71-targeting nanotherapeutic named The-0504 on preclinical models of several human aggressive tumors.

J Exp Clin Cancer Res 2021 Feb 10;40(1):63. Epub 2021 Feb 10.

CNR - National Research Council of Italy, Institute of Molecular Biology and Pathology, Rome, Italy.

Background: Ferritin receptor (CD71) is an example of a very attractive cancer target, since it is highly expressed in virtually all tumor types, including metastatic loci. However, this target can be considered to be inaccessible to conventional target therapies, due to its presence in many healthy tissues. Here, we describe the preclinical evaluation of a tumor proteases-activatable human ferritin (HFt)-based drug carrier (The-0504) that is able to selectively deliver the wide-spectrum topoisomerase I inhibitor Genz-644282 to CD71-expressing tumors, preventing the limiting toxic effects associated with CD71-targeting therapies.

Methods: CD71 expression was evaluated using flow cytometry and immunohistochemistry techniques. The-0504 antiproliferative activity towards several cancer cell lines was assessed in vitro. The-0504 antitumor efficacy and survival benefit were evaluated in different human tumors, which had been grown either as xenografts or patient-derived xenografts in mice. The-0504 toxicology profile was investigated in multiple-cycle repeat-dose study in rodents.

Results: In vitro studies indicate that The-0504 is highly specific for CD71 expressing cells, and that there is a relationship between CD71 levels and The-0504 anticancer activity. In vivo treatments with The-0504 showed a remarkable efficacy, eradicating several human tumors of very diverse and aggressive histotypes, such as pancreas, liver and colorectal carcinomas, and triple-negative breast cancer.

Conclusions: Durable disease-free survival, persistent antitumor responses after discontinuation of treatment and favorable toxicology profile make The-0504 an ideal candidate for clinical development as a novel, CD71-targeted, low-toxicity alternative to chemotherapy.
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http://dx.doi.org/10.1186/s13046-021-01851-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7877078PMC
February 2021

Immunolocalization of leptin and leptin receptor in colorectal mucosa of ulcerative colitis, Crohn's disease and control subjects with no inflammatory bowel disease.

Cell Tissue Res 2021 Mar 7;383(3):1103-1122. Epub 2020 Nov 7.

Department of Neuroscience, Biomedicine and Movement, Human Anatomy and Histology Section, University of Verona, 37134, Verona, Italy.

The expression of leptin and leptin receptor (Ob-R) has been partially elucidated in colon of patients with inflammatory bowel diseases (IBDs), even though leptin is involved in angiogenesis and inflammation. We previously reported overexpression of GLUT5 fructose transporter, in aberrant clusters of lymphatic vessels in lamina propria of IBD and controls. Here, we examine leptin and Ob-R expression in the same biopsies. Specimens were obtained from patients with ulcerative colitis (UC), Crohn's disease (CD) and controls who underwent screening for colorectal cancer, follow-up after polypectomy or with a history of lower gastrointestinal symptoms. Immunohistochemistry revealed leptin in apical and basolateral membranes of short epithelial portions, Ob-R on the apical pole of epithelial cells. Leptin and Ob-R were also identified in structures and cells scattered in the lamina propria. In UC, a significant correlation between leptin and Ob-R in the lamina propria was found in all inflamed samples, beyond non-inflamed samples of the proximal tract, while in CD, it was found in inflamed distal samples. Most of the leptin and Ob-R positive areas in the lamina propria were also GLUT5 immunoreactive in inflamed and non-inflamed mucosa. A significant correlation of leptin or Ob-R expression with GLUT5 was observed in the inflamed distal samples from UC. Our findings suggest that there are different sites of leptin and Ob-R expression in large intestine and those in lamina propria do not reflect the status of mucosal inflammation. The co-localization of leptin and/or Ob-R with GLUT5 may indicate concomitance effects in colorectal lamina propria areas.
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http://dx.doi.org/10.1007/s00441-020-03297-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7960629PMC
March 2021

Nanoparticles for Cerenkov and Radioluminescent Light Enhancement for Imaging and Radiotherapy.

Nanomaterials (Basel) 2020 Sep 7;10(9). Epub 2020 Sep 7.

Experimental Imaging Center, San Raffaele Scientific Institute, Via Olgettina 60, 20132 Milan, Italy.

Cerenkov luminescence imaging and Cerenkov photodynamic therapy have been developed in recent years to exploit the Cerenkov radiation (CR) generated by radioisotopes, frequently used in Nuclear Medicine, to diagnose and fight cancer lesions. For in vivo detection, the endpoint energy of the radioisotope and, thus, the total number of the emitted Cerenkov photons, represents a very important variable and explains why, for example, Ga is better than F. However, it was also found that the scintillation process is an important mechanism for light production. Nanotechnology represents the most important field, providing nanosctructures which are able to shift the UV-blue emission into a more suitable wavelength, with reduced absorption, which is useful especially for in vivo imaging and therapy applications. Nanoparticles can be made, loaded or linked to fluorescent dyes to modify the optical properties of CR radiation. They also represent a useful platform for therapeutic agents, such as photosensitizer drugs for the production of reactive oxygen species (ROS). Generally, NPs can be spaced by CR sources; however, for in vivo imaging applications, NPs bound to or incorporating radioisotopes are the most interesting nanocomplexes thanks to their high degree of mutual colocalization and the reduced problem of false uptake detection. Moreover, the distance between the NPs and CR source is crucial for energy conversion. Here, we review the principal NPs proposed in the literature, discussing their properties and the main results obtained by the proponent experimental groups.
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http://dx.doi.org/10.3390/nano10091771DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559269PMC
September 2020

Hedonicity in functional motor disorders: a chemosensory study assessing taste.

J Neural Transm (Vienna) 2020 10 28;127(10):1399-1407. Epub 2020 Aug 28.

Department of Neurosciences, Biomedicine and Movement Sciences, Neurology Section, Verona University Hospital, University of Verona, Piazzale Scuro 10, 37134, Verona, Italy.

The aim of this study was to explore hedonicity to basic tastes in patients with functional motor disorders (FMDs) that are often associated with impairment in emotional processing. We recruited 20 FMD patients and 24 healthy subjects, matched for age and sex. Subjects were asked to rate the hedonic sensation (i.e., pleasant, neutral, and unpleasant) on a - 10 to +10 scale to the four basic tastes (sweet, sour, salty, and bitter) at different concentrations, and neutral stimuli (i.e., no taste stimulation) by means of the Taste Strips Test. Anxiety, depression, and alexithymia were assessed. FMD patients rated the highest concentration of sweet taste (6.7 ± 2.6) as significantly more pleasant than controls (4.7 ± 2.5, p = 0.03), and the neutral stimuli significantly more unpleasant (patients: - 0.7 ± 0.4, controls: 0.1 ± 0.4, p = 0.013). Hedonic ratings were not correlated to anxiety, depression, or alexithymia scores. Hedonic response to taste is altered in FMD patients. This preliminary finding might result from abnormal interaction between sensory processing and emotional valence.
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http://dx.doi.org/10.1007/s00702-020-02244-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7497316PMC
October 2020

Disabled Homolog 2 Controls Prometastatic Activity of Tumor-Associated Macrophages.

Cancer Discov 2020 Nov 10;10(11):1758-1773. Epub 2020 Jul 10.

Department of Medicine, Section of Immunology, University of Verona, Verona, Italy.

Tumor-associated macrophages (TAM) are regulators of extracellular matrix (ECM) remodeling and metastatic progression, the main cause of cancer-associated death. We found that disabled homolog 2 mitogen-responsive phosphoprotein (DAB2) is highly expressed in tumor-infiltrating TAMs and that its genetic ablation significantly impairs lung metastasis formation. DAB2-expressing TAMs, mainly localized along the tumor-invasive front, participate in integrin recycling, ECM remodeling, and directional migration in a tridimensional matrix. DAB2 macrophages escort the invasive dissemination of cancer cells by a mechanosensing pathway requiring the transcription factor YAP. In human lobular breast and gastric carcinomas, DAB2 TAMs correlated with a poor clinical outcome, identifying DAB2 as potential prognostic biomarker for stratification of patients with cancer. DAB2 is therefore central for the prometastatic activity of TAMs. SIGNIFICANCE: DAB2 expression in macrophages is essential for metastasis formation but not primary tumor growth. Mechanosensing cues, activating the complex YAP-TAZ, regulate DAB2 in macrophages, which in turn controls integrin recycling and ECM remodeling in 3-D tissue matrix. The presence of DAB2 TAMs in patients with cancer correlates with worse prognosis..
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http://dx.doi.org/10.1158/2159-8290.CD-20-0036DOI Listing
November 2020

A Correlative Imaging Study of in vivo and ex vivo Biodistribution of Solid Lipid Nanoparticles.

Int J Nanomedicine 2020 13;15:1745-1758. Epub 2020 Mar 13.

Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Verona I-37134, Italy.

Purpose: Solid lipid nanoparticles are largely used in biomedical research and are characterized by high stability and biocompatibility and are also able to improve the stability of various loaded molecules. In vitro studies demonstrated that these nanoparticles are low cytotoxic, while in vivo studies proved their efficiency as nanocarriers for molecules characterized by a low bioavailability. However, to our knowledge, no data on the systemic biodistribution and organ accumulation of solid lipid nanoparticles in itself are presently available.

Methods: In this view, we investigated the solid lipid nanoparticles biodistribution by a multimodal imaging approach correlating in vivo and ex vivo analyses. We loaded solid lipid nanoparticles with two different fluorophores (cardiogreen and rhodamine) to observe them with an optical imager in the whole organism and in the excised organs, and with fluorescence microscopy in tissue sections. Light and transmission electron microscopy analyses were also performed to evaluate possible structural modification or damage due to nanoparticle administration.

Results: Solid lipid nanoparticles loaded with the two fluorochromes showed good optic characteristics and stable polydispersity. After in vivo administration, they were clearly detectable in the organism. Four  hours after the injection, the fluorescent signal occurred in anatomical districts corresponding to the liver and this was confirmed by the ex vivo acquisitions of excised organs. Brightfield, fluorescence and transmission electron microscopy confirmed solid lipid nanoparticles accumulation in hepatocytes without structural damage.

Conclusion: Our results support the systemic biocompatibility of solid lipid nanoparticles and demonstrate their detailed biodistribution from the whole organism to organs until the cells.
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http://dx.doi.org/10.2147/IJN.S236968DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7078788PMC
July 2020

Proteomic and Ultrastructural Analysis of Cellulite-New Findings on an Old Topic.

Int J Mol Sci 2020 03 18;21(6). Epub 2020 Mar 18.

Department of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology division, University of Verona, 37134 Verona, Italy.

Background: Cellulite is a condition in which the skin has a dimpled lumpy appearance. The main causes of cellulite development, studied until now, comprehends modified sensitivity to estrogens, the damage of microvasculature present among dermis and hypodermis. The differences of adipose tissue architecture between male and female might make female more susceptible to cellulite. Adipose tissue is seen to be deeply modified during cellulite development. Our study tried to understand the overall features within and surrounding cellulite to apply the best therapeutic approach.

Methods: Samples of gluteal femoral area were collected from cadavers and women who had undergone surgical treatment to remove orange peel characteristics on the skin. Samples from cadavers were employed for an accurate study of cellulite using magnetic resonance imaging at 7 Tesla and for light microscopy. Specimens from patients were employed for the proteomic analysis, which was performed using high resolution mass spectroscopy (MS). Stromal vascular fraction (SVF) was obtained from the samples, which was studied using MS and flow cytometry.

Results: light and electron microscopy of the cellulite affected area showed a morphology completely different from the other usual adipose depots. In cellulite affected tissues, sweat glands associated with adipocytes were found. In particular, there were vesicles in the extracellular matrix, indicating a crosstalk between the two different components. Proteomic analysis showed that adipose tissue affected by cellulite is characterized by high degree of oxidative stress and by remodeling phenomena.

Conclusions: The novel aspects of this study are the peculiar morphology of adipose tissue affected by cellulite, which could influence the surgical procedures finalized to the reduction of dimpling, based on the collagen fibers cutting. The second novel aspect is the role played by the mesenchymal stem cells isolated from stromal vascular fraction of adipose tissue affected by cellulite.
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http://dx.doi.org/10.3390/ijms21062077DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7139738PMC
March 2020

Tandem Dye-Doped Nanoparticles for NIR Imaging via Cerenkov Resonance Energy Transfer.

Front Chem 2020 27;8:71. Epub 2020 Feb 27.

Department of Chemistry "Giacomo Ciamician", University of Bologna, Bologna, Italy.

The detection of the Cerenkov radiation (CR) is an emerging preclinical imaging technique which allows monitoring the distribution of radionuclides. Among its possible advantages, the most interesting is the simplicity and cost of the required instrumentation compared, e.g., to that required for PET scans. On the other hand, one of its main drawbacks is related to the fact that CR, presenting the most intense component in the UV-vis region, has a very low penetration in biological tissues. To address this issue, we present here multifluorophoric silica nanoparticles properly designed to efficiently absorb the CR radiation and to have a quite high fluorescence quantum yield (0.12) at 826 nm. Thanks to a highly efficient series of energy transfer processes, each nanoparticle can convert part of the CR into NIR light, increasing its detection even under 1.0-cm thickness of muscle.
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http://dx.doi.org/10.3389/fchem.2020.00071DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7056810PMC
February 2020

Small animal irradiator dose distribution verification using radioluminescence imaging.

J Biophotonics 2020 07 7;13(7):e201960217. Epub 2020 Apr 7.

Department of Computer Science, University of Verona, Italy.

The main objective of this work was the development of a novel 2D dosimetry approach for small animal external radiotherapy using radioluminescence imaging (RLI) with a commercial complementary metal oxide semiconductor detector. Measurements of RLI were performed on the small animal image-guided platform SmART, RLI data were corrected for perspective distortion using Matlab. Four irradiation fields were tested and the planar 2D dose distributions and dose profiles were compared against dose calculations performed with a Monte Carlo based treatment planning system and gafchromic film. System linearity and RLI image noise against dose were also measured. The maximum difference between beam size measured with RLI and nominal beam size was less than 8% for all the tested beams. The image correction procedure was able to reduce perspective distortion. A novel RLI approach for quality assurance of a small animal irradiator was presented and tested. Results are in agreement with MC dose calculations and gafchromic film measurements.
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http://dx.doi.org/10.1002/jbio.201960217DOI Listing
July 2020

Comparison of the Effects of Browning-Inducing Capsaicin on Two Murine Adipocyte Models.

Front Physiol 2019 5;10:1380. Epub 2019 Nov 5.

Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy.

The increasing prevalence of obesity and its associated comorbidities has gained attention in developing effective treatments and strategies that promote energy expenditure and the conversion of fat from a white to a brite phenotype. Capsaicin, bioactive component of chili peppers and a transient receptor potential channel vanilloid 1 (TRPV1) agonist, has been known to stimulate the process of thermogenesis. In this study, the effects of capsaicin were assessed on two murine cellular models by quantifying the dynamic of lipid droplets (LDs) and the expression of genes involved in adipocyte browning. Present findings demonstrated that treatment with norepinephrine or capsaicin combined with norepinephrine on 3T3-L1 cells and X9 cells significantly promoted the reduction of LDs area surface and size. The transcription of browning related genes such as uncoupling protein 1 (), T-box transcription factor 1 (), PR domain containing 16 (), peroxisome proliferator-activated receptor γ coactivator 1α () and cell death-inducing DNA fragmentation factor A-like effector A () was up-regulated by chronic capsaicin treatment on differentiated 3T3-L1 cells. Instead, X9 cells were significantly responsive only to the treatment with norepinephrine, used as positive control.
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http://dx.doi.org/10.3389/fphys.2019.01380DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6848400PMC
November 2019

Modulating TAK1 Expression Inhibits YAP and TAZ Oncogenic Functions in Pancreatic Cancer.

Mol Cancer Ther 2020 01 27;19(1):247-257. Epub 2019 Sep 27.

Digestive Molecular Clinical Oncology Research Unit, Department of Medicine, Università degli studi di Verona, Verona, Italy.

YAP and TAZ are central determinants of malignancy; however, their functions remain still undruggable. We identified TGFβ-activated kinase 1 (TAK1) as a central hub integrating the most relevant signals sustaining pancreatic cancer aggressiveness and chemoresistance. Glycogen synthase kinase (GSK)3 is known to stabilize TAK1, and its inhibition causes a reduction in TAK1 levels. Here, we hypothesized that TAK1 could sustain YAP/TAZ program, and thus, modulation of TAK1 expression through the inhibition of GSK3 could impair YAP/TAZ functions in pancreatic cancer.Differentially expressed transcripts between pancreatic cancer cells expressing scramble or -specific shRNA were annotated for functional interrelatedness by ingenuity pathway analysis. TAK1 expression was modulated by using different GSK3 inhibitors, including LY2090314. activity of LY2090314 alone or in combination with nab-paclitaxel was evaluated in an orthotopic nude mouse model.Differential gene expression profiling revealed significant association of TAK1 expression with HIPPO and ubiquitination pathways. We measured a significant downregulation of YAP/TAZ and their regulated genes in shTAK1 cells. TAK1 prevented YAP/TAZ proteasomal degradation in a kinase independent manner, through a complex with TRAF6, thereby fostering their K63-ubiquitination versus K48-ubiquitination. Pharmacologic modulation of TAK1 by using GSK3 inhibitors significantly decreased YAP/TAZ levels and suppressed their target genes and oncogenic functions. , LY2090314 plus nab-paclitaxel significantly prolonged mice survival duration.Our study demonstrates a unique role for TAK1 in controlling YAP/TAZ in pancreatic cancer. LY2090314 is a novel agent that warrants further clinical development in combination with nab-paclitaxel for the treatment of pancreatic cancer.
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http://dx.doi.org/10.1158/1535-7163.MCT-19-0270DOI Listing
January 2020

Weak biophoton emission after laser surgery application in soft tissues: Analysis of the optical features.

J Biophotonics 2019 09 14;12(9):e201800260. Epub 2019 Jun 14.

Experimental Imaging Centre, San Raffaele Scientific Institute, Milan, Italy.

Nowadays, laser scalpels are commonly used in surgery, replacing the traditional surgical scalpels for several applications involving cutting or ablating living biological tissue. Laser scalpels are generally used to concentrate light energy in a very small-sized area; light energy is then converted in heat by the tissues. In other cases, the fiber glass tip of the laser scalpel is heated to high temperature and used to cut the tissues. Depending on the temperature reached in the irradiated area, different effects are visible in the tissues. In this study, we report the discovery and characterization of the light emitted by soft mammalian biological tissues from seconds to hours after laser surgery application. A laser diode (with hot fiber glass tip) working at 808 nm and commercially available for medical and dentistry applications was used. The irradiated tissues (red meat, chicken breast and fat) showed light emission in the visible range, well detectable with a commercial charge coupled device (CCD) camera. The time decay of the light emission, the laser power effects and the spectral features in the range 500 to 840 nm in the different tissues are here reported.
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http://dx.doi.org/10.1002/jbio.201800260DOI Listing
September 2019

Relationship between lipid droplets size and integrated optical density.

Eur J Histochem 2019 Mar 25;63(1). Epub 2019 Mar 25.

University of Verona, Department of Computer Science.

Lipid accumulation is largely investigated due to its role in many human diseases. The attention is mainly focused on the lipid droplets (LDs), spherical cytoplasmic organelles, which are devoted to the storage of the lipids. The amount of lipid content is often evaluated by measuring LDs size and/or the integrated optical density (IOD) in cultured cells. Both evaluations are directly associated to the lipid content and therefore they are correlated to each other, but a lack of theoretical relationship between size and IOD was observed in literature. Here we investigated the size-IOD relationship of LDs observed in microscopical images of cultured cells. The experimental data were obtained from immature and differentiated 3T3-L1 murine cells, which have been extensively used in studies on adipogenesis. A simple model based on the spherical shape of the LDs and the Lambert-Beer law, which describes the light absorption by an optical thick material, leads to a mathematical relationship. Despite only light rays' absorption was considered in the model, neglecting their scattering, a very good agreement between the theoretical curve and the experimental data was found. Moreover, a computational simulation corroborates the model indicating the validity of the mathematically theoretical relationship between size and IOD. The theoretical model could be used to calculate the absorption coefficient in the LDs population and it could be applied to seek for morphologically and functionally LDs subpopulations. The identification of LDs dynamic by measuring size and IOD could be related to different pathophysiological conditions and useful for understand cellular lipid-associated diseases.
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http://dx.doi.org/10.4081/ejh.2019.3017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6444373PMC
March 2019

Uptake and intracellular fate of biocompatible nanocarriers in cycling and noncycling cells.

Nanomedicine (Lond) 2019 02 22;14(3):301-316. Epub 2019 Jan 22.

Department of Neurosciences, Biomedicine & Movement Sciences, University of Verona, Strada Le Grazie, 8 - 37134 Verona, Italy.

Aim: To elucidate whether different cytokinetic features (i.e., presence or absence of mitotic activity) may influence cell uptake and distribution of nanocarriers, in vitro tests on liposomes, mesoporous silica nanoparticles, poly(lactide-co-glycolide) nanoparticles and nanohydrogels were carried out on C2C12 murine muscle cells either able to proliferate as myoblasts (cycling cells) or terminally differentiate into myotubes (noncycling cells).

Materials & Methods: Cell uptake and intracellular fate of liposomes, mesoporous silica nanoparticles, poly(lactide-co-glycolide) nanoparticles and nanohydrogels were investigated by confocal fluorescence microscopy and transmission electron microscopy.

Results: Nanocarrier internalization and distribution were similar in myoblasts and myotubes; however, myotubes demonstrated a lower uptake capability.

Conclusion: All nanocarriers proved to be suitably biocompatible for both myoblasts and myotubes. The lower uptake capability of myotubes is probably due to different plasma membrane composition related to the differentiation process.
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http://dx.doi.org/10.2217/nnm-2018-0148DOI Listing
February 2019

Dynamic of lipid droplets and gene expression in response to β-aminoisobutyric acid treatment on 3T3-L1 cells.

Eur J Histochem 2018 Nov 28;62(4). Epub 2018 Nov 28.

University of Udine, Department of Agricultural, Food, Environmental and Animal Sciences.

Research on adipobiology has recognized the browning process of white adipocytes as a potential therapeutic strategy for the treatment of obesity and related morbidities. Physical exercise stimulates the secretion of myokines, such as b-aminoisobutyric acid (BAIBA), which in turn promotes adaptive thermogenesis. White adipocyte conversion to brown cells involves dynamic changes in lipid droplet (LD) dimension and in the transcription of brown-specific marker genes. This study analyzes the effect of different doses of BAIBA and at different days of development on 3T3-L1 cells by evaluating morphological changes in LDs and the expression of browning gene markers. Results suggested that the highest concentration of BAIBA after 4 days of differentiation produced the most significant effects. The number of LDs per cell increased in comparison to control cells, whereas the surface area significantly decreased. Brown adipocyte markers were up-regulated, but the effect of treatment was lost at 10 days of differentiation. The thermogenic program induced by BAIBA may reflect a rapid adaptation of adipose tissue to physical exercise. This connection stresses the beneficial impact of physical exercise on metabolic health. The thermogenic program induced by BAIBA may reflect a rapid adaptation of adipose tissue to physical exercise. This connection stresses the beneficial impact of physical exercise on metabolic health.
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http://dx.doi.org/10.4081/ejh.2018.2984DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6280065PMC
November 2018

Ozone Treatment of Grapes During Withering for Amarone Wine: A Multimodal Imaging and Spectroscopic Analysis.

Microsc Microanal 2018 10;24(5):564-573

1Department of Neurosciences,Biomedicine and Movement Sciences,University of Verona,Strada Le Grazie 8,Verona 37134,Italy.

The production of Amarone wine is governed by a disciplinary guideline to preserve its typical features; however, postharvest infections by the fungus Botrytis cinerea (B. cinerea) not only represent a phytosanitary problem but also cause a significant loss of product. In this study, we tested a treatment with mild ozoniztion on grapes for Amarone wine production during withering in the fruttaio (the environment imposed by the disciplinary guideline) and evaluated the impact on berry features by a multimodal imaging approach. The results indicate that short and repeated treatments with low O3 concentrations speed up the naturally occurring berry withering, probably inducing a reorganization of the epicuticular wax layer, and inhibit the development of B. cinerea, blocking the fungus in an intermediate vegetative stage. This pilot study will pave the way to long-term research on Amarone wine obtained from O3-treated grapes.
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http://dx.doi.org/10.1017/S1431927618015209DOI Listing
October 2018

Low ozone concentrations promote adipogenesis in human adipose-derived adult stem cells.

Eur J Histochem 2018 Sep 3;62(3). Epub 2018 Sep 3.

University of Verona, Department of Neurosciences, Biomedicine and Movement Sciences.

Ozone is a strong oxidant, highly unstable atmospheric gas. Its medical use at low concentrations has been progressively increasing as an alternative/adjuvant treatment for several diseases. In this study, we investigated the effects of mild ozonisation on human adipose-derived adult stem (hADAS) cells i.e., mesenchymal stem cells occurring in the stromal-vascular fraction of the fat tissue and involved in the tissue regeneration processes. hADAS cells were induced to differentiate into the adipoblastic lineage, and the effect of low ozone concentrations on the adipogenic process was studied by combining histochemical, morphometric and ultrastructural analyses. Our results demonstrate that ozone treatment promotes lipid accumulation in hADAS without inducing deleterious effects, thus paving the way to future studies aimed at elucidating the effect of mild ozonisation on adipose tissue for tissue regeneration and engineering.
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http://dx.doi.org/10.4081/ejh.2018.2969DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6151336PMC
September 2018

The role of mutated SOD1 gene in synaptic stripping and MHC class I expression following nerve axotomy in ALS murine model.

Eur J Histochem 2018 May 17;62(2):2904. Epub 2018 May 17.

University of Kentucky Medical Center.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by motoneuron death. Several cellular pathways have been described to be involved in ALS pathogenesis; however, the involvement of presynaptic stripping and the related MHC class I molecules in mutant SOD1 motoneurons remains to be clarified. To this purpose, we here investigated, for the first time, the motoneurons behavior, di per seand after facial axonal injury, in terms of synaptic stripping and MHC class I expression in wild-type (Wt) mice and in a murine model of ALS, the SOD1(G93A) mice, at the presymptomatic and symptomatic stage of the disease. Concerning Wt animals, we found a reduction in synaptophysin immunoreactivity and an increase of MHC class I molecules in facial motoneurons after axotomy. In uninjured motoneurons of SOD1(G93A) mice, an altered presynaptic framework was evident, and this phenomenon increased during the disease course. The alteration in the presynaptic input is related to excitatory fibers. Moreover, after injury, a further decrease of excitatory input was not associated to an upregulation of MHC class I molecules in motoneuron soma. This study demonstrates, for the first time, that the presence of mutated SOD1 protein affects the MHC class I molecules expression, altering the presynaptic input in motoneurons. Nevertheless, a positive MHC class I immunolabeling was evident in glial cells around facial injured motoneurons, underlying an involvement of these cells in synaptic stripping. This study contributes to better understand the involvement of the mutated SOD1 protein in the vulnerability of motoneurons after damage.
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http://dx.doi.org/10.4081/ejh.2018.2904DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5966710PMC
May 2018

Inhibition of Pseudomonas aeruginosa secreted virulence factors reduces lung inflammation in CF mice.

Virulence 2018 ;9(1):1008-1018

a Department of Diagnostics and Public Health , University of Verona , Verona , Italy.

Background: Cystic fibrosis (CF) lung infection is a complex condition where opportunistic pathogens and defective immune system cooperate in developing a constant cycle of infection and inflammation. The major pathogen, Pseudomonas aeruginosa, secretes a multitude of virulence factors involved in host immune response and lung tissue damage. In this study, we examined the possible anti-inflammatory effects of molecules inhibiting P. aeruginosa virulence factors.

Methods: Pyocyanin, pyoverdine and proteases were measured in bacterial culture supernatant from different P. aeruginosa strains. Inhibition of virulence factors by sub-inhibitory concentrations of clarithromycin and by protease inhibitors was evaluated. Lung inflammatory response was monitored by in vivo bioluminescence imaging in wild-type and CFTR-knockout mice expressing a luciferase gene under the control of a bovine IL-8 promoter.

Results: The amount of proteases, pyocyanin and pyoverdine secreted by P. aeruginosa strains was reduced after growth in the presence of a sub-inhibitory dose of clarithromycin. Intratracheal challenge with culture supernatant containing bacteria-released products induced a strong IL-8-mediated response in mouse lungs while lack of virulence factors corresponded to a reduction in bioluminescence emission. Particularly, sole inactivation of proteases by inhibitors Ilomastat and Marimastat also resulted in decreased lung inflammation.

Conclusions: Our data support the assumption that virulence factors are involved in P. aeruginosa pro-inflammatory action in CF lungs; particularly, proteases seem to play an important role. Inhibition of virulence factors production and activity resulted in decreased lung inflammation; thus, clarithromycin and protease inhibitors potentially represent additional therapeutic therapies for P. aeruginosa-infected patients.
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http://dx.doi.org/10.1080/21505594.2018.1489198DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6086295PMC
October 2018

Mild ozonisation activates antioxidant cell response by the Keap1/Nrf2 dependent pathway.

Free Radic Biol Med 2018 08 2;124:114-121. Epub 2018 Jun 2.

Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Strada Le Grazie 8, I-37134 Verona, Italy. Electronic address:

Treatment with low-dose ozone is successfully exploited as an adjuvant therapy in the treatment of several disorders. Although the list of medical applications of ozone therapy is increasing, molecular mechanisms underlying its beneficial effects are still partially known. Clinical and experimental evidence suggests that the therapeutic effects of ozone treatment may rely on its capability to mount a beneficial antioxidant response through activation of the nuclear factor erythroid-derived-like 2 (Nrf2) pathway. However, a conclusive mechanistic demonstration is still lacking. Here, we bridge this gap of knowledge by providing evidence that treatment with a low concentration of ozone in cultured cells promotes nuclear translocation of Nrf2 at the chromatin sites of active transcription and increases the expression of antioxidant response element (ARE)-driven genes. Importantly, we show that ozone-induced ARE activation can be reverted by the ectopic expression of the Nrf2 specific inhibitor Kelch-like ECH associated protein (Keap1), thus proving the role of the Nrf2 pathway in the antioxidant response induced by mild ozonisation.
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http://dx.doi.org/10.1016/j.freeradbiomed.2018.05.093DOI Listing
August 2018

T-cell tracking using Cerenkov and radioluminescence imaging.

J Biophotonics 2018 10 5;11(10):e201800093. Epub 2018 Jul 5.

Experimental Imaging Centre, San Raffaele Scientific Institute, Milan, Italy.

Cancer immunotherapy is a promising strategy based on the ability of the immune system to kill selected cells. In the development of an effective T-cell therapy, the noninvasive cell tracking methods play a crucial role. Here, we investigate the potentialities of T-cell marked with radionuclides in order to detect their localization with imaging techniques in small animal rodents. A protocol to label T-cells with P-ATP was tested and evaluated. The homing of P-ATP labeled T lymphocytes was investigated by Cerenkov luminescence imaging (CLI) and radioluminescence imaging (RLI). The first approach relies on the acquisition of Cerenkov photons produced by the beta particles emitted by the P internalized by lymphocytes; the second one on the detection of photons coming from the conversion of radioactive energy in light done by scintillator crystals layered on the animals. The results show that T-cell biodistribution can be optically observed by both CLI and RLI in small animal rodents in in vivo and ex vivo acquisitions. T-cell localization in the tumor mass was definitively confirmed by flow cytometry.
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http://dx.doi.org/10.1002/jbio.201800093DOI Listing
October 2018

Weak light emission of soft tissues induced by heating.

J Biomed Opt 2018 04;23(4):1-5

University of Verona, Department of Computer Science, Verona, Italy.

The main goal of this work is to show that soft tissue interaction with high-intensity focused ultrasound (HIFU) or direct heating leads to a weak light emission detectable using a small animal optical imaging system. Our results show that the luminescence signal is detectable after 30 min of heating, resembling the time scale of delayed luminescence. The imaging of a soft tissue after heating it using an HIFU field shows that the luminescence pattern closely matches the shape of the cone typical of the HIFU beam. We conclude that heating a soft tissue using two different sources leads to the emission of a weak luminescence signal from the heated region with a decay half-life of a few minutes (4 to 6 min). The origin of such light emission needs to be further investigated.
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http://dx.doi.org/10.1117/1.JBO.23.4.046003DOI Listing
April 2018

Glucose transporter expression in the human colon.

World J Gastroenterol 2018 Feb;24(7):775-793

Department of Surgery, Oncology and Gastroenterology, Gastroenterology Section, University Hospital of Padua, Padua I-35128, Italy.

Aim: To investigate by immunostaining glucose transporter expression in human colorectal mucosa in controls and patients with inflammatory bowel disease (IBD).

Methods: Colorectal samples were obtained from patients undergoing lower endoscopic colonoscopy or recto-sigmoidoscopy. Patients diagnosed with ulcerative colitis ( = 18) or Crohn's disease ( = 10) and scheduled for diagnostic colonoscopy were enrolled. Patients who underwent colonoscopy for prevention screening of colorectal cancer or were followed-up after polypectomy or had a history of lower gastrointestinal symptoms were designated as the control group (CTRL, = 16). Inflammatory status of the mucosa at the sampling site was evaluated histologically and/or endoscopically. A total of 147 biopsies of colorectal mucosa were collected and processed for immunohistochemistry analysis. The expression of GLUT2, SGLT1, and GLUT5 glucose transporters was investigated using immunoperoxidase labeling. To compare immunoreactivity of GLUT5 and LYVE-1, which is a marker for lymphatic vessel endothelium, double-labeled confocal microscopy was used.

Results: Immunohistochemical analysis revealed that GLUT2, SGLT1, and GLUT5 were expressed only in short epithelial portions of the large intestinal mucosa. No important differences were observed in glucose transporter expression between the samples obtained from the different portions of the colorectal tract and between the different patient groups. Unexpectedly, GLUT5 expression was also identified in vessels, mainly concentrated in specific areas where the vessels were clustered. Immunostaining with LYVE-1 and GLUT5 antibodies revealed that GLUT5-immunoreactive (-IR) clusters of vessels were concentrated in areas internal to those that were LYVE-1 positive. GLUT5 and LYVE-1 did not appear to be colocalized but rather showed a close topographical relationship on the endothelium. Based on their LYVE-1 expression, GLUT5-IR vessels were identified as lymphatic. Both inflamed and non-inflamed mucosal colorectal tissue biopsies from the IBD and CTRL patients showed GLUT5-IR clusters of lymphatic vessels.

Conclusion: Glucose transporter immunoreactivity is present in colorectal mucosa in controls and IBD patients. GLUT5 expression is also associated with lymphatic vessels. This novel finding aids in the characterization of lymphatic vasculature in IBD patients.
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http://dx.doi.org/10.3748/wjg.v24.i7.775DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5807937PMC
February 2018

Hyaluronated mesoporous silica nanoparticles for active targeting: influence of conjugation method and hyaluronic acid molecular weight on the nanovector properties.

J Colloid Interface Sci 2018 Apr 31;516:484-497. Epub 2018 Jan 31.

Department of Drug Science and Technology, University of Torino, Via P. Giuria 9, Torino, Italy. Electronic address:

We have prepared and evaluated the physico-chemical and biological properties of four different hyaluronated mesoporous silica nanoparticles (MSNs) samples (MSN/HA). Hyaluronic acid (HA) with two different molecular weights (200 and 6.4 kDa) was used for the conjugation of aminopropyl-functionalized MSN (NH-MSN), following two different procedures. Namely, samples HA200A and HA6.4A were prepared by reacting activated HA with NH-MSN (method A), while samples HA200B and HA6.4B were obtained carrying out HA activation in the presence of the nanoparticles (method B). The four samples showed similar hydrophilicity, but clear differences in the HA loading, textural properties, surface charge and stability of the suspensions. More in detail, conjugation using low molecular weight HA with method A resulted in low HA loading, with consequent scarce effects on dispersity and stability in physiological media. The highest yield and corresponding best performances were obtained with method B using high molecular weight HA. HA loading and molecular weight also influenced in a concerted way the biological response towards the MSNs of CD44 target cancer cells (CD44) and control cells (CD44): MDA-MB-231 and A2780, respectively. The absence of cytotoxicity was assessed. Moreover, the targeting ability of the best performing MSN/HA was confirmed by cellular uptake studies.
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http://dx.doi.org/10.1016/j.jcis.2018.01.072DOI Listing
April 2018

Metabolic effect of bodyweight whole-body vibration in a 20-min exercise session: A crossover study using verified vibration stimulus.

PLoS One 2018 31;13(1):e0192046. Epub 2018 Jan 31.

Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Verona, Italy.

The ability of whole body vibration (WBV) to increase energy expenditure (EE) has been investigated to some extent in the past using short-term single exercises or sets of single exercises. However, the current practice in WBV training for fitness is based on the execution of multiple exercises during a WBV training session for a period of at least 20 min; nevertheless, very limited and inconsistent data are available on EE during long term WBV training session. This crossover study was designed to demonstrate, in an adequately powered sample of participants, the ability of WBV to increase the metabolic cost of exercise vs. no vibration over the time span of a typical WBV session for fitness (20 min). Twenty-two physically active young males exercised on a vibration platform (three identical sets of six different exercises) using an accelerometer-verified vibration stimulus in both the WBV and no vibration condition. Oxygen consumption was measured with indirect calorimetry and expressed as area under the curve (O2(AUC)). Results showed that, in the overall 20-min training session, WBV increased both the O2(AUC) and the estimated EE vs. no vibration by about 22% and 20%, respectively (P<0.001 for both, partial eta squared [η2] ≥0.35) as well as the metabolic equivalent of task (+5.5%, P = 0.043; η2 = 0.02) and the rate of perceived exertion (+13%, P<0.001; ŋ2 = 0.16). Results demonstrated that vibration is able to significantly increase the metabolic cost of exercise in a 20-min WBV training session.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0192046PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5792008PMC
March 2018

Radioluminescence from Tc-99m in glass predicts local dose.

Phys Med 2017 Oct 15;42:112-115. Epub 2017 Sep 15.

San Raffaele Scientific Institute, Experimental Imaging Center, Via Olgettina N. 60, Milan 20182, Italy.

The widely-used gamma-emitter Tc-99m has been shown to lead to optical emissions in mice and glass. We investigated the possibility that these emissions are due to the Cerenkov effect and whether the light emitted is proportional to local dose. By using a Geant4 Monte Carlo model matched to an experimental measurement, we show that the light detected by a small animal optical imaging system provides a 2D map of the dose throughout a glass sample. We conclude that radioluminescence from Tc-99m can be used to quantitatively measure dose in transparent materials, which could have applications in dosimetry and quality assurance.
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http://dx.doi.org/10.1016/j.ejmp.2017.09.119DOI Listing
October 2017

Theranostic Role of P-ATP as Radiopharmaceutical for the Induction of Massive Cell Death within Avascular Tumor Core.

Theranostics 2017 30;7(18):4399-4409. Epub 2017 Sep 30.

Experimental Imaging Centre, San Raffaele Scientific Institute, Via Olgettina 60, 20182 Milan, Italy.

Drug inaccessibility to vast areas of the tumor parenchyma is amongst the major hurdles for conventional therapies. Treatment efficacy rapidly decreases with distance from vessels and most of the tumor cells survive therapy. Also, between subsequent cycles of treatment, spared cancer cells replace those killed near the vessels, improving their access to nutrients, boosting their proliferation rate, and thus enabling tumor repopulation. Because of their property of "acting at a distance," radioisotopes are believed to overcome the physical barrier of vascular inaccessibility. A novel molecular imaging tool called Cerenkov Luminescence Imaging (CLI) was employed for the detection of Cerenkov radiation emitted by beta particles, allowing tracking of beta-emitters. More precisely we investigated using a xenograft model of colon carcinoma the potential use of 32P-ATP as a novel theranostic radiopharmaceutical for tracing tumor lesions while simultaneously hampering their growth. Our analyses demonstrated that 32P-ATP injected into tumor-bearing mice reaches tumor lesions and persists for days and weeks within the tumor parenchyma. Also, the high-penetrating beta particles of 32P-ATP exert a "cross-fire" effect that induces massive cell death throughout the entire tumor parenchyma including core regions. Our findings suggest 32P-ATP treatment as a potential approach to complement conventional therapies that fail to reach the tumor core and to prevent tumor repopulation.
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http://dx.doi.org/10.7150/thno.21403DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5695139PMC
March 2018

Effective control of acute myeloid leukaemia and acute lymphoblastic leukaemia progression by telomerase specific adoptive T-cell therapy.

Oncotarget 2017 Oct 23;8(50):86987-87001. Epub 2017 May 23.

Department of Medicine, University of Verona, Section of Immunology, Verona, Italy.

Telomerase (TERT) is a ribonucleoprotein enzyme that preserves the molecular organization at the ends of eukaryotic chromosomes. Since TERT deregulation is a common step in leukaemia, treatments targeting telomerase might be useful for the therapy of hematologic malignancies. Despite a large spectrum of potential drugs, their bench-to-bedside translation is quite limited, with only a therapeutic vaccine in the clinic and a telomerase inhibitor at late stage of preclinical validation. We recently demonstrated that the adoptive transfer of T cell transduced with an HLA-A2-restricted T-cell receptor (TCR), which recognize human TERT with high avidity, controls human B-cell chronic lymphocytic leukaemia (B-CLL) progression without severe side-effects in humanized mice. In the present report, we show the ability of our approach to limit the progression of more aggressive leukemic pathologies, such as acute myeloid leukaemia (AML) and B-cell acute lymphoblastic leukaemia (B-ALL). Together, our findings demonstrate that TERT-based adoptive cell therapy is a concrete platform of T cell-mediated immunotherapy for leukaemia treatment.
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http://dx.doi.org/10.18632/oncotarget.18115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5675610PMC
October 2017

Innovative approach to safely induce controlled lipolysis by superparamagnetic iron oxide nanoparticles-mediated hyperthermic treatment.

Int J Biochem Cell Biol 2017 12 27;93:62-73. Epub 2017 Oct 27.

Dipartimento di Neuroscienze, Biomedicina e Movimento, Università di Verona, 37134 Verona, Italy. Electronic address:

During last years, evidence has been provided on the involvement of overweight and obesity in the pathogenesis and aggravation of several life-threatening diseases. Here, we demonstrate that, under appropriate administration conditions, polyhedral iron oxide nanoparticles are efficiently and safely taken up by 3T3 cell line-derived adipocytes (3T3 adipocytes) in vitro. Since these nanoparticles proved to effectively produce heat when subjected to alternating magnetic field, 3T3 adipocytes were submitted to superparamagnetic iron oxide nanoparticles-mediated hyperthermia treatment (SMHT), with the aim of modulating their lipid content. Notably, the treatment resulted in a significant delipidation persisting for at least 24h, and in the absence of cell death, damage or dedifferentiation. Interestingly, transcript expression of adipose triglyceride lipase (ATGL), a key gene involved in canonical lipolysis, was not modulated upon SMHT, suggesting the involvement of a novel/alternative mechanism in the effective lipolysis observed. By applying the same experimental conditions successfully used for 3T3 adipocytes, SMHT was able to induce delipidation also in primary cultures of human adipose-derived adult stem cells. The success of this pioneering approach in vitro opens promising perspectives for the application of SMHT in vivo as an innovative safe and physiologically mild strategy against obesity, potentially useful in association with balanced diet and healthy lifestyle.
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http://dx.doi.org/10.1016/j.biocel.2017.10.013DOI Listing
December 2017