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Article ; Online: Incubation of canine dermal fibroblasts with serum from dogs with atopic dermatitis activates extracellular matrix signalling and represses oxidative phosphorylation.

Colitti, Monica / Stefanon, Bruno / Sandri, Misa / Licastro, Danilo

2022 Volume 47, Issue 1, Page(s) 247–258

Abstract: The aim of this study was to investigate the effects on gene expression in canine fibroblasts after incubation with a medium enriched with atopic dermatitis canine serum (CAD) compared with healthy canine serum (CTRL) and fetal bovine serum (FBS). ... ...

Abstract	The aim of this study was to investigate the effects on gene expression in canine fibroblasts after incubation with a medium enriched with atopic dermatitis canine serum (CAD) compared with healthy canine serum (CTRL) and fetal bovine serum (FBS). Differential Expression and Pathway analysis (iDEP94) in R package (v0.92) was used to identify differentially expressed genes (DEGs) with a False Discovery Rate of 0.01. DEGs from fibroblasts incubated with CAD serum were significantly upregulated and enriched in the extracellular matrix (ECM) and focal adhesion signalling but downregulated in the oxidative phosphorylation pathway. Genes involved in profibrotic processes, such as TGFB1, INHBA, ERK1/2, and the downward regulated genes (collagens and integrins), were significantly upregulated after fibroblasts were exposed to CAD serum. The observed downregulation of genes involved in oxidative phosphorylation suggests metabolic dysregulation toward a myofibroblast phenotype responsible for fibrosis. No differences were found when comparing CTRL with FBS. The DEGs identified in fibroblasts incubated with CAD serum suggest activation of signalling pathways involved in gradual differentiation through a myofibroblast precursors that represent the onset of fibrosis. Molecular and metabolic knowledge of fibroblast changes can be used to identify biomarkers of the disease and new potential pharmacological targets.
MeSH term(s)	Dogs ; Animals ; Dermatitis, Atopic/veterinary ; Dermatitis, Atopic/metabolism ; Dermatitis, Atopic/pathology ; Oxidative Phosphorylation ; Fibroblasts ; Extracellular Matrix ; Fibrosis ; Dog Diseases/pathology
Language	English
Publishing date	2022-06-04
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	406735-6
ISSN	1573-7446 ; 0165-7380
ISSN (online)	1573-7446
ISSN	0165-7380
DOI	10.1007/s11259-022-09947-y
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Article ; Online: Incubation of canine dermal fibroblasts with serum from dogs with atopic dermatitis activates extracellular matrix signalling and represses oxidative phosphorylation

Colitti, Monica / Stefanon, Bruno / Sandri, Misa / Licastro, Danilo

Vet Res Commun. 2023 Jan., v. 47, no. 1 p.247-258

2023

Abstract	The aim of this study was to investigate the effects on gene expression in canine fibroblasts after incubation with a medium enriched with atopic dermatitis canine serum (CAD) compared with healthy canine serum (CTRL) and fetal bovine serum (FBS). Differential Expression and Pathway analysis (iDEP94) in R package (v0.92) was used to identify differentially expressed genes (DEGs) with a False Discovery Rate of 0.01. DEGs from fibroblasts incubated with CAD serum were significantly upregulated and enriched in the extracellular matrix (ECM) and focal adhesion signalling but downregulated in the oxidative phosphorylation pathway. Genes involved in profibrotic processes, such as TGFB1, INHBA, ERK1/2, and the downward regulated genes (collagens and integrins), were significantly upregulated after fibroblasts were exposed to CAD serum. The observed downregulation of genes involved in oxidative phosphorylation suggests metabolic dysregulation toward a myofibroblast phenotype responsible for fibrosis. No differences were found when comparing CTRL with FBS. The DEGs identified in fibroblasts incubated with CAD serum suggest activation of signalling pathways involved in gradual differentiation through a myofibroblast precursors that represent the onset of fibrosis. Molecular and metabolic knowledge of fibroblast changes can be used to identify biomarkers of the disease and new potential pharmacological targets.
Keywords	atopic dermatitis ; biomarkers ; dogs ; extracellular matrix ; fetal bovine serum ; fibroblasts ; fibrosis ; focal adhesions ; gene expression ; gene expression regulation ; integrins ; oxidative phosphorylation ; phenotype
Language	English
Dates of publication	2023-01
Size	p. 247-258.
Publishing place	Springer Netherlands
Document type	Article ; Online
ZDB-ID	406735-6
ISSN	1573-7446 ; 0165-7380
ISSN (online)	1573-7446
ISSN	0165-7380
DOI	10.1007/s11259-022-09947-y
Database	NAL-Catalogue (AGRICOLA)

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Article: Regulatory Role of microRNA of Milk Exosomes in Mastitis of Dairy Cows.

Stefanon, Bruno / Cintio, Michela / Sgorlon, Sandy / Scarsella, Elisa / Licastro, Danilo / Zecconi, Alfonso / Colitti, Monica

Animals : an open access journal from MDPI

2023 Volume 13, Issue 5

Abstract: The aim of this study was to compare the cargos of miRNA in exosomes isolated from the milk of healthy (H) cows, cows at risk of mastitis (ARM), and cows with subclinical mastitis (SCM). Based on the number of somatic cells and the percentage of ... ...

Abstract	The aim of this study was to compare the cargos of miRNA in exosomes isolated from the milk of healthy (H) cows, cows at risk of mastitis (ARM), and cows with subclinical mastitis (SCM). Based on the number of somatic cells and the percentage of polymorphonuclear cells, 10 cows were assigned to group H, 11 to group ARM, and 11 to group SCM. After isolating exosomes in milk by isoelectric precipitation and ultracentrifugation, the extracted RNA was sequenced to 50 bp long single reads, and these were mapped against Btau_5.0.1. The resulting 225 miRNAs were uploaded to the miRNet suite, and target genes for
Language	English
Publishing date	2023-02-24
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani13050821
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Article ; Online: Regulatory Role of microRNA of Milk Exosomes in Mastitis of Dairy Cows

Bruno Stefanon / Michela Cintio / Sandy Sgorlon / Elisa Scarsella / Danilo Licastro / Alfonso Zecconi / Monica Colitti

Animals, Vol 13, Iss 821, p

2023 Volume 821

Abstract	The aim of this study was to compare the cargos of miRNA in exosomes isolated from the milk of healthy (H) cows, cows at risk of mastitis (ARM), and cows with subclinical mastitis (SCM). Based on the number of somatic cells and the percentage of polymorphonuclear cells, 10 cows were assigned to group H, 11 to group ARM, and 11 to group SCM. After isolating exosomes in milk by isoelectric precipitation and ultracentrifugation, the extracted RNA was sequenced to 50 bp long single reads, and these were mapped against Btau_5.0.1. The resulting 225 miRNAs were uploaded to the miRNet suite, and target genes for Bos taurus were identified based on the miRTarBase and miRanda databases. The list of differentially expressed target genes resulting from the comparisons of the three groups was enriched using the Function Explorer of the Kyoto Encyclopedia of Genes and Genomes. A total of 38, 18, and 12 miRNAs were differentially expressed (DE, p < 0.05) in the comparisons of H vs. ARM, ARM vs. SCM, and H vs. SCM, respectively. Only 1 DE miRNA was shared among the three groups (bta-mir-221), 1 DE miRNA in the H vs. SCM comparison, 9 DE miRNAs in the ARM vs. SCM comparison, and 21 DE miRNAs in the H vs. ARM comparison. A comparison of the enriched pathways of target genes from the H, SCM, and ARM samples showed that 19 pathways were differentially expressed in the three groups, while 56 were expressed in the H vs. SCM comparison and 57 in the H vs. ARM comparison. Analyzing milk exosome miRNA cargos can be considered as a promising approach to study the complex molecular machinery set in motion in response to mastitis in dairy cows.
Keywords	milk ; exosomes ; miRNA ; mastitis ; cow ; Veterinary medicine ; SF600-1100 ; Zoology ; QL1-991
Subject code	630
Language	English
Publishing date	2023-02-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Elemental composition in commercial dry extruded and moist canned dog foods

Sgorlon, Sandy / Sandri, Misa / Stefanon, Bruno / Licastro, Danilo

Animal feed science and technology. 2022 May, v. 287

2022

Abstract: The aim of the study was to investigate elemental contents in commercial pet foods for adult dogs at maintenance. The elemental composition of 15 dry extruded diets (DED) and 22 moist canned diets (MCD) collected from a pet shop were analyzed in ... ...

Abstract	The aim of the study was to investigate elemental contents in commercial pet foods for adult dogs at maintenance. The elemental composition of 15 dry extruded diets (DED) and 22 moist canned diets (MCD) collected from a pet shop were analyzed in triplicate with inductively coupled plasma optical emission spectroscopy and with mass spectrometry for Se. Significantly higher contents of Ca, P, (P < 0.05), K and Na (P < 0.0001) were measured in MCD than in DED, whilst the contrary was observed for Cu and Zn (P = 0.001). All the analyzed foods supplied on average micro minerals above the recommended daily requirements of FEDIAF, except for Cu (mean supply of foods 85%+12) in the MCD and Se both in DED (mean supply in foods 21%+4) and MCD (mean supply in foods 34%+7) samples. All the analyzed foods supplied macro elements above the recommended daily allowances except for K (73%+6) in the DED and Mg (92%+5) in MCD. Principal component analysis with essential elements (EE) as input variables (Ca, P, K, Na, Mg, Fe, Mn, Zn, Cu, Se) explained 63.85% of total variance with the first two components and clustered DED and MCD. Factor scores of Cu, Zn, Mg and Mn were higher for MCD and Ca, P, K and Na for DED. Significantly higher (P < 0.01) frequency of presence of non essential elements (NEE) Cd, Cr, Li and Pb (p < 0.01) were observed in MCD in comparison to DED. Although there are limitations in this study, the results showed a wide variability of elemental contents among per foods and suggest the need of routinely analysis to accomplish with the recommended daily allowances and for the safeguard of companion animals health.
Keywords	adults ; dogs ; elemental composition ; mass spectrometry ; principal component analysis ; technology ; variance
Language	English
Dates of publication	2022-05
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	196563-3
ISSN	0377-8401
ISSN	0377-8401
DOI	10.1016/j.anifeedsci.2022.115287
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Regulatory Role of microRNA of Milk Exosomes in Mastitis of Dairy Cows

Stefanon, Bruno / Cintio, Michela / Sgorlon, Sandy / Scarsella, Elisa / Licastro, Danilo / Zecconi, Alfonso / Colitti, Monica

Animals. 2023 Feb. 24, v. 13, no. 5

2023

Abstract	The aim of this study was to compare the cargos of miRNA in exosomes isolated from the milk of healthy (H) cows, cows at risk of mastitis (ARM), and cows with subclinical mastitis (SCM). Based on the number of somatic cells and the percentage of polymorphonuclear cells, 10 cows were assigned to group H, 11 to group ARM, and 11 to group SCM. After isolating exosomes in milk by isoelectric precipitation and ultracentrifugation, the extracted RNA was sequenced to 50 bp long single reads, and these were mapped against Btau_5.0.1. The resulting 225 miRNAs were uploaded to the miRNet suite, and target genes for Bos taurus were identified based on the miRTarBase and miRanda databases. The list of differentially expressed target genes resulting from the comparisons of the three groups was enriched using the Function Explorer of the Kyoto Encyclopedia of Genes and Genomes. A total of 38, 18, and 12 miRNAs were differentially expressed (DE, p < 0.05) in the comparisons of H vs. ARM, ARM vs. SCM, and H vs. SCM, respectively. Only 1 DE miRNA was shared among the three groups (bta-mir-221), 1 DE miRNA in the H vs. SCM comparison, 9 DE miRNAs in the ARM vs. SCM comparison, and 21 DE miRNAs in the H vs. ARM comparison. A comparison of the enriched pathways of target genes from the H, SCM, and ARM samples showed that 19 pathways were differentially expressed in the three groups, while 56 were expressed in the H vs. SCM comparison and 57 in the H vs. ARM comparison. Analyzing milk exosome miRNA cargos can be considered as a promising approach to study the complex molecular machinery set in motion in response to mastitis in dairy cows.
Keywords	cattle ; chemical precipitation ; exosomes ; gene expression regulation ; genome ; mastitis ; microRNA ; milk ; risk ; ultracentrifugation
Language	English
Dates of publication	2023-0224
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article ; Online
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani13050821
Database	NAL-Catalogue (AGRICOLA)

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Article: A mutation in the ZNF687 gene that is responsible for the severe form of Paget's disease of bone causes severely altered bone remodeling and promotes hepatocellular carcinoma onset in a knock-in mouse model.

Russo, Sharon / Scotto di Carlo, Federica / Maurizi, Antonio / Fortunato, Giorgio / Teti, Anna / Licastro, Danilo / Settembre, Carmine / Mello, Tommaso / Gianfrancesco, Fernando

Bone research

2023 Volume 11, Issue 1, Page(s) 16

Abstract: Paget's disease (PDB) is a late-onset bone remodeling disorder with a broad spectrum of symptoms and complications. One of the most aggressive forms is caused by the P937R mutation in the ZNF687 gene. Although the genetic involvement of ZNF687 in PDB has ...

Abstract	Paget's disease (PDB) is a late-onset bone remodeling disorder with a broad spectrum of symptoms and complications. One of the most aggressive forms is caused by the P937R mutation in the ZNF687 gene. Although the genetic involvement of ZNF687 in PDB has been extensively studied, the molecular mechanisms underlying this association remain unclear. Here, we describe the first Zfp687 knock-in mouse model and demonstrate that the mutation recapitulates the PDB phenotype, resulting in severely altered bone remodeling. Through microcomputed tomography analysis, we observed that 8-month-old mutant mice showed a mainly osteolytic phase, with a significant decrease in the trabecular bone volume affecting the femurs and the vertebrae. Conversely, osteoblast activity was deregulated, producing disorganized bone. Notably, this phenotype became pervasive in 16-month-old mice, where osteoblast function overtook bone resorption, as highlighted by the presence of woven bone in histological analyses, consistent with the PDB phenotype. Furthermore, we detected osteophytes and intervertebral disc degeneration, outlining for the first time the link between osteoarthritis and PDB in a PDB mouse model. RNA sequencing of wild-type and Zfp687 knockout RAW264.7 cells identified a set of genes involved in osteoclastogenesis potentially regulated by Zfp687, e.g., Tspan7, Cpe, Vegfc, and Ggt1, confirming its role in this process. Strikingly, in this mouse model, the mutation was also associated with a high penetrance of hepatocellular carcinomas. Thus, this study established an essential role of Zfp687 in the regulation of bone remodeling, offering the potential to therapeutically treat PDB, and underlines the oncogenic potential of ZNF687.
Language	English
Publishing date	2023-03-14
Publishing country	China
Document type	Journal Article
ZDB-ID	2803313-9
ISSN	2095-6231 ; 2095-4700
ISSN (online)	2095-6231
ISSN	2095-4700
DOI	10.1038/s41413-023-00250-3
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Link between organic nanovescicles from vegetable kingdom and human cell physiology: intracellular calcium signalling.

Trentini, Martina / Zanolla, Ilaria / Tiengo, Elena / Zanotti, Federica / Sommella, Eduardo / Merciai, Fabrizio / Campiglia, Pietro / Licastro, Danilo / Degasperi, Margherita / Lovatti, Luca / Bonora, Massimo / Danese, Alberto / Pinton, Paolo / Zavan, Barbara

Journal of nanobiotechnology

2024 Volume 22, Issue 1, Page(s) 68

Abstract: Background: Plant-derived nanovesicles (PDNVs) are a novelty in medical and agrifood environments, with several studies exploring their functions and potential applications. Among fruits, apples (sp. Malus domestica) have great potential as PDNVs source, ...

Abstract	Background: Plant-derived nanovesicles (PDNVs) are a novelty in medical and agrifood environments, with several studies exploring their functions and potential applications. Among fruits, apples (sp. Malus domestica) have great potential as PDNVs source, given their widespread consumption, substantial waste production, and recognized health benefits. Notably, apple-derived nanovesicles (ADNVs) can interact with human cell lines, triggering anti-inflammatory and antioxidant responses. This work is dedicated to the comprehensive biochemical characterization of apple-derived nanovesicles (ADNVs) through proteomic and lipidomic analysis, and small RNAs sequencing. This research also aims to shed light on the underlying mechanism of action (MOA) when ADNVs interface with human cells, through observation of intracellular calcium signalling in human fibroblasts, and to tackles differences in ADNVs content when isolated from fruits derived from integrated and organic production methods cultivars. Results: The ADNVs fraction is mainly composed of exocyst-positive organelles (EXPOs) and MVB-derived exosomes, identified through size and molecular markers (Exo70 and TET-3-like proteins). ADNVs' protein cargo is heterogeneous and exhibits a diverse array of functions, especially in plant's protection (favouring ABA stress-induced signalling, pathogen resistance and Reactive Oxygen Species (ROS) metabolism). Noteworthy plant miRNAs also contribute to phytoprotection. In relation with human cells lines, ADNVs elicit spikes of intracellular Ca Conclusions: We have conclusively demonstrated that ADNVs confer an antioxidant effect upon human cells, through the initiation of a molecular pathway triggered by Ca
MeSH term(s)	Humans ; Antioxidants/pharmacology ; Antioxidants/metabolism ; Calcium/metabolism ; Vegetables ; Proteomics ; Malus/metabolism ; Signal Transduction
Chemical Substances	Antioxidants ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2024-02-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2100022-0
ISSN	1477-3155 ; 1477-3155
ISSN (online)	1477-3155
ISSN	1477-3155
DOI	10.1186/s12951-024-02340-8
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Adipose-derived stem cell exosomes act as delivery vehicles of microRNAs in a dog model of chronic hepatitis.

Zanolla, Ilaria / Trentini, Martina / Tiengo, Elena / Zanotti, Federica / Pusceddu, Tommaso / Rubini, Andrea / Rubini, Giuseppe / Brugnoli, Francesca / Licastro, Danilo / Debortoli, Marco / Delogu, Lucia Gemma / Ferroni, Letizia / Lovatti, Luca / Zavan, Barbara

Nanotheranostics

2024 Volume 8, Issue 3, Page(s) 298–311

Abstract: Exosomes are nanosized extracellular vesicles secreted by all cell types, including canine adipose-derived stem cells (cADSCs). By mediating intercellular communication, exosomes modulate the biology of adjacent and distant cells by transferring their ... ...

Abstract	Exosomes are nanosized extracellular vesicles secreted by all cell types, including canine adipose-derived stem cells (cADSCs). By mediating intercellular communication, exosomes modulate the biology of adjacent and distant cells by transferring their cargo. In the present work after isolation and characterization of exosomes derived from canine adipose tissue, we treated the same canine donors affected by hepatopathies with the previously isolated exosomes. We hypothesize that cADSC-sourced miRNAs are among the factors responsible for a regenerative and anti-inflammatory effect in the treatment of hepatopathies in dogs, providing the clinical veterinary field with an effective and innovative cell-free therapy. Exosomes were isolated and characterized for size, distribution, surface markers, and for their miRNomic cargo by microRNA sequencing. 295 dogs affected with hepatopathies were treated and followed up for 6 months to keep track of their biochemical marker levels. Results confirmed that exosomes derived from cADSCs exhibited an average diameter of 91 nm, and positivity to 8 known exosome markers. The administration of exosomes to dogs affected by liver-associated inflammatory pathologies resulted in the recovery of the animal alongside the normalization of biochemical parameters of kidney function. In conclusion, cADSCs-derived exosomes are a promising therapeutic tool for treating inflammatory disorders in animal companions.
MeSH term(s)	Dogs ; Animals ; MicroRNAs/genetics ; Exosomes/genetics ; Exosomes/metabolism ; Extracellular Vesicles/metabolism ; Hepatitis, Chronic/metabolism ; Stem Cells/metabolism
Chemical Substances	MicroRNAs
Language	English
Publishing date	2024-03-09
Publishing country	Australia
Document type	Journal Article
ISSN	2206-7418
ISSN (online)	2206-7418
DOI	10.7150/ntno.93064
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Genome Sequences of SARS-CoV-2 Strains from the Republic of Moldova.

Ulinici, Mariana / Soñora, Martín / Orsini, Emanuele / Licastro, Danilo / Dal Monego, Simeone / Todiras, Mihail / Lungu, Ludmila / Groppa, Stanislav / Marcello, Alessandro

Microbiology resource announcements

2022 Volume 12, Issue 1, Page(s) e0113222

Abstract: The whole-genome sequences of 15 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains from nasopharyngeal swab samples collected in the Republic of Moldova in June 2020 to September 2021 were determined. Little variability was observed in ...

Abstract	The whole-genome sequences of 15 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains from nasopharyngeal swab samples collected in the Republic of Moldova in June 2020 to September 2021 were determined. Little variability was observed in the early stages, when mostly clade 19A was circulating, followed by clade 20B. Later, multiple introductions of SARS-CoV-2 lineages B.1.1., B.1.1.7, and B.1.1.525 were detected. The B.1.1.7 lineage became predominant between December 2020 and June 2021, followed by the Delta variant.
Language	English
Publishing date	2022-12-12
Publishing country	United States
Document type	Journal Article
ISSN	2576-098X
ISSN (online)	2576-098X
DOI	10.1128/mra.01132-22
Database	MEDical Literature Analysis and Retrieval System OnLINE

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