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Central giant cell granulomas (CGCG) are rare intraosseous osteolytic lesions of uncertain aetiology. Despite the benign nature of this neoplasia, the lesions can rapidly grow and become large, painful, invasive, and destructive. The identification of molecular drivers could help in the selection of targeted therapies for specific cases. TRPV4, KRAS and FGFR1 mutations have been associated with these lesions but no correlation between the mutations and patient features was observed so far. In this study, we analysed 17 CGCG cases of an Italian cohort and identified an interesting and significant (p=0.0021) correlation between FGFR1 mutations and age. In detail, FGFR1 mutations were observed frequently and exclusively in CGCG from young (18 years old) patients (4/5 lesions, 80%). Furthermore, the combination between ours and previously published data confirmed a significant difference in the frequency of FGFR1 mutations in CGCG from patients younger than 18 years at the time of diagnosis (9/23 lesions, 39%) when compared to older patients (1/31 lesions, 0.03%; p=0.0011), thus corroborating our observation in a cohort of 54 patients. FGFR1 variants in young CGCG patients could favour fast lesion growth, implying that they seek medical attention earlier. Our observation might help prioritise candidates for FGFR1 testing, thus opening treatment options with FGFR inhibitors.

Analytical advancements in lipidomics have enabled large-scale investigations of lipid biology. Herein, we focused on four bioactive lipid families, namely polyunsaturated fatty acids, eicosanoids, endocannabinoids, and N-acylethanolamines, and their involvement in the mesenchymal stem cells (MSC)-related inflammatory scenario. Since MSC secretome may represent a valid therapeutic alternative, here, the complete secretome and its vesicular component from adipose- and bone marrow-derived MSC and dermal fibroblasts were characterized by targeted mass spectrometry lipidomics. The 2-arachidonoylglycerol (2AG) and the palmitoylethanolamide (PEA), previously quantified in the MSC's secretome, were further investigated by assessing hypothetical effects in an in vitro model of osteoarthritis (OA) based on human primary articular chondrocytes (CH) stimulated with tumor necrosis factor alpha (TNFα). TNFα enhances the release of the inflammatory lipid prostaglandin E2 (PGE2), and an additional increment was observed when CH were treated with both TNFα and 2AG. In contrast, PEA downmodulates the PGE2 release to the levels of unstimulated CH suggesting a protective effect. TNFα also increases the expression of cyclooxygenase 2 (COX2), in particular when combined with 2AG, while PEA partly blunts TNFα-induced COX2 expression. In addition, TNFα-stimulated CH produce significantly higher levels of the inflammatory mediator nitric oxide (NO) both in the presence and in the absence of 2AG, and PEA was able to partially reduce NO release. Our results show a first partial lipidomic profile of MSC and DF secretome and suggest a possible implication of bioactive lipids in the OA scenario and in the future use of these cell-free products as innovative therapeutics.

Human adipose-derived stem/stromal cells (hASCs) have been popularly studied as cell-based therapy in the field of regenerative medicine due to their ability to differentiate into several cell types. In this study, in order to improve the mechanical strength and bioactivity of scaffolds for bone tissue engineering, three types of mesoporous bioactive glasses with different shapes and compositions were dispersed in the silk fibroin/chitosan (SF/CS)-based scaffolds, which were fabricated with a combination of freezing and lyophilization. The characteristic and physical properties of these composite scaffolds were evaluated. The biocompatibility was also assessed through hASCs in vitro tests. Both Alamar Blue® and Live/Dead assay® revealed that the spherical mesoporous bioactive glass doped scaffolds enhanced cell viability and proliferation. Furthermore, the addition of spherical mesoporous bioactive glass into SF/CS scaffolds encouraged hASC osteogenic differentiation as well. These results suggested that this composite scaffold can be applicable material for bone regeneration. &nbsp

BACKGROUND Metastatic disease is frequently present at the time of diagnosis of canine thyroid carcinoma; however, utilisation of computed tomography (CT) alone for staging pre-treatment has been rarely reported in the veterinary literature. METHODS The aims of this retrospective study were to stage affected dogs using CT findings of the cervical and thoracic regions, combined with histopathology/cytology results, in order to assess whether metastatic disease/WHO staging was of prognostic significance. RESULTS Fifty-eight dogs were included in the study. Classification of cases into WHO stages I, II, III and IV were 10%, 50%, 9% and 31%, respectively. No statistically significant effect of WHO stage classification on overall survival/follow-up time was found (P = .576). Surgery resulted in a statistically significant increase in overall survival/follow-up time (P < .01). There was no statistically significant effect on overall survival/follow-up time in dogs that received medical therapy, either as sole therapy or as an adjunctive post-surgery (P = .198). CONCLUSION In summary, this study documents the metastatic rate of canine thyroid carcinoma using CT for staging pre-treatment. Staging utilising CT revealed a higher distant metastatic rate in dogs with thyroid carcinoma when compared to historical studies using different imaging techniques. As long-term outcomes are possible for cases with advanced disease, surgical intervention could still be considered.

Changes in lipid metabolism are involved in several pathological conditions, such as cancer. Among lipids, eicosanoids are potent inflammatory mediators, synthesized from polyunsaturated fatty acids (PUFAs), which coexist with other lipid-derived ones, including endocannabinoids (ECs) and N-acylethanolamides (NAEs). In this work, a bioanalytical assay for 12 PUFAs/eicosanoids and 20 ECs/NAEs in cell culture medium and human biofluids was validated over a linear range of 0.1&ndash, 2.5 ng/mL. A fast pretreatment method consisting of protein precipitation with acetonitrile followed by a double step liquid&ndash, liquid extraction was developed. The final extracts were injected onto a Kinetex ultra-high-performance liquid chromatography (UHPLC) XB-C18 column with a gradient elution of 0.1% formic acid in water and methanol/acetonitrile (5:1, v/v) mobile phase. Chromatographic separation was followed by detection with a triple-quadrupole mass spectrometer operating both in positive and negative ion-mode. A full validation was carried out in a small amount of cell culture medium and then applied to osteosarcoma cell-derived products. To the best of our knowledge, this is the first lipid profiling of bone tumor cell lines (SaOS-2 and MG-63) and their secretome. Our method was also partially validated in other biological matrices, such as serum and urine, ensuring its broad applicability as a powerful tool for lipidomic translational research.

Over the years, several studies have shown that many factors are likely to affect the results of forensic hair analyses and complicate their interpretation. Among these factors, one of the major drawbacks in hair analysis is the affectability of deposited xenobiotics by cosmetic treatments, which could be eventually used to adulterate the sample. It is well known that some cosmetic treatments containing hydrogen peroxide, such as permanent dyeing or bleaching, lead to the formation of 1-H-pyrrole-2,3,5-tricarboxylic acid (PTCA), a melanin degradation product. Considering that PTCA is also an endogenous compound, spontaneously formed by natural oxidation of melanin, its only detection in hair is not enough to confirm a cosmetic oxidative treatment. For this reason, the aim of the present work was to develop and validate a reliable liquid–liquid extraction method in ultra-high-performance liquid chromatographic–tandem mass spectrometry for the determination of endogenous PTCA in hair from a wide multi-ethnic population (African, Arab, Asian-Pacific, Caucasian, Hispanic and Indian). According to previous studies, untreated hair samples showed a PTCA content of 8.54 ± 5.72 ng/mg (mean ± standard deviation [SD]), ranging between 0.44 and 23.7 ng/mg; after in vitro cosmetic bleaching, PTCA increased to 16.8 ± 6.95 ng/mg (range: 4.16–32.3 ng/mg). Comparing baseline PTCA levels of each subgroup with the others, we could not observe any statistically significant difference, except for Caucasians (P

Case summaryIn this report we describe the occurrence of intracranial meningioma in two adult cats from the same litter. The location of the meningioma varied: one tumour was at the level of the brainstem, and the other was affecting the temporal and piriform lobes. The cat with the brainstem meningioma was treated with radiotherapy and the littermate had a rostrotentorial craniectomy for tumour removal. Both cats had a histopathological diagnosis of grade I meningioma of a predominantly fibrous subtype.Relevance and novel informationCases of familial meningioma in cats have not previously been described in the veterinary literature. However, familial meningioma is well described in humans and it is possible that cases are underestimated in animals. We discuss the possible genetic background and other causes, as well as challenges we may face in veterinary medicine in identifying these associations.

Conditioned medium (CM) and extracellular vesicles (EV) from Adipose-derived Stem/stromal cells (ASC) and Dermal fibroblasts (DF) represent promising tools for therapeutic applications. Which one should be preferred is still under debate and no direct comparison of their proteome has been reported yet. Here, we apply quantitative proteomics to explore the protein composition of CM and EV from the two cell types. Data are available via ProteomeXchange (identifier PXD020219). We identified 1977 proteins by LC-MS/MS proteomic analysis. Unsupervised clustering analysis and PCA recognized CM and EV as separate groups. We identified 68 and 201 CM and EV specific factors. CM were enriched in proteins of endoplasmic reticulum, Golgi apparatus and lysosomes, whereas EV contained a large amount of GTPases, ribosome and translation factors. The analysis of ASC and DF secretomes revealed the presence of cell type-specific proteins. ASC-CM and -EV carried factors involved in ECM organization and immunological regulation, respectively. Conversely, DF-CM and -EV were enriched in epithelium development associated factors and -EV in Wnt signaling factors. In conclusion, this analysis provides evidence of a different protein composition between CM and EV and of the presence of cell type-specific bioactive mediators suggesting their specific future use as advanced therapy medicinal products. SIGNIFICANCE: The use of cell secretome presents several advantages over cell therapy such as the lower risks associated to the administration step and the avoidance of any potential risk of malignant transformation. The main secretome preparations consist in concentrated conditioned medium (CM) and extracellular vesicles (EV). Both of them showed well-documented therapeutic potentials. However, it is still not clear in which case it should be better to use one preparation over the other and an exhaustive comparison between their proteome has not been performed yet. The choice of the cell source is another relevant aspect that still needs to be addressed. In order to shed light on these questions we explored the protein composition of CM and EV obtained from Adipose-derived Stem/stromal Cells (ASC) and Dermal Fibroblasts (DF), by a comprehensive quantitative proteomics approach. The analysis showed a clear distinction between CM and EV proteome. CM were enriched in proteins of endoplasmic reticulum, Golgi apparatus and lysosomes, whereas EV contained a large amount of GTPases, ribosome and translation-related factors. Furthermore, the analysis of ASC and DF secretomes revealed specific biological processes for the different cell products. ASC secretome presented factors involved in ECM organization (hyaluronan and glycosaminoglycan metabolism) and immunological regulation (e.g. macrophage and IkB/NFkB signaling regulation), respectively. On the other hand, DF-CM and -EV were both enriched in epithelium development associated factors, whilst DF-CM in proteins involved in cellular processes regulation and -EV in Wnt signaling factors. In conclusion, our study shed a light on the different protein composition of CM and EV of two promising cell types, spanning from basic processes involved in secretion to specific pathways supporting their therapeutic potential and their possible future use as advanced therapy medicinal products.

Background A previous study showed an association between owner-reported exposure to environmental tobacco smoke (ETS) and lymphoma in cats. This study aimed to investigate the association between ETS exposure and gastrointestinal lymphoma in cats, using hair nicotine concentration (HNC) as a biomarker. Methods This was a prospective, multi-centre, case–control study. Gastrointestinal lymphoma was diagnosed on cytology or histopathology. Hair samples were obtained from 35 cats with gastrointestinal lymphoma and 32 controls. Nicotine was extracted from hair by sonification in methanol followed by hydrophilic interaction chromatography with mass spectrometry. Non-parametric tests were used. Results The median HNC of the gastrointestinal lymphoma and control groups was not significantly different (0.030 ng/mg and 0.029 ng/mg, respectively, p=0.46). When the HNC of all 67 cats was rank ordered and divided into quartiles, there was no significant difference in the proportion of lymphoma cases or controls within these groups (p=0.63). The percentage of cats with an HNC≥0.1 ng/mg was higher for the lymphoma group (22.9%) than the control group (15.6%) but failed to reach significance (p=0.45). Conclusion A significant association was not identified between HNC (a biomarker for ETS) and gastrointestinal lymphoma in cats; however, an association may exist and further studies are therefore required.

An 8-year-old female neutered Australian Cattle Dog that had recently moved to the UK from Arizona presented with a 4-month history of lethargy, inappetance and right forelimb lameness, and a 6-month history of stranguria and haematuria. Diagnosis and treatment of a fungal infection are described.

A 5-year-old Domestic Shorthair cat with a history of IMHA (successfully treated a year before) presented as an emergency following a tonic clonic seizure. The diagnosis and treatment of polycythaemia are described.

OBJECTIVE: Gingival tissue is composed of cell types that contribute to the body's defense against many agents in oral environment, wound healing and tissue regeneration. Thanks to their easy and scarcely invasive withdrawal procedure, interdental papilla provide a good source of mesenchymal stromal cells (GinPa-MSCs). We isolated GinPa-MSCs and verified their ability to uptake/release the anticancer agent Paclitaxel (PTX). METHODS: In vitro expanded GinPa-MSCs were characterized for CD markers by FACS, tested for differentiation ability and analyzed by TEM. Their ability to uptake/release PTX was assessed according to a standardized procedure. RESULTS: The CD expression and chondro-adipo-osteo differentiation ability confirmed the mesenchymal feature of GinPa-MSCs. Surprisingly, 28% of GinPa-MSCs expressed CD14 marker and had an impressive pinocytotic activity. GinPa-MSCs were able to take up and release a sufficient amount of PTX to demonstrate effective in vitro activity against pancreatic carcinoma cells, suggesting that the drug was not inactivated. CONCLUSIONS: The procedure to obtain MSCs from interdental papilla is less invasive than that used for both bone marrow and adipose tissue, GinPa-MSCs are easy to expand and can be efficiently loaded with PTX. Taken together these qualities suggest that GinPa-MSCs may prove to be a good tool for cell-mediated drug delivery in cancer, particularly if related to stomatognathic system.

In the last few years, great advances have been made in exploring the clinical potential of oral and dental stem cells. Indeed, in dentistry, the restoration of tooth and periodontal tissues is now focusing on the use of Mesenchymal Stromal Cells (MSCs) of oral origins, such as tooth pulp (DPSCs), periodontal ligament (PDLSCs), dental follicle (DFPCs) and apical papilla (SCAPs). In particular, thanks to the easy and minimally invasive withdrawal procedure, oral soft tissues represent a convenient source of MSCs. Gingival tissue is composed by a variety of cell types and in the oral environment it represents the first important barrier against external offensive insults (e.g. chemicals, viruses and bacteria); another peculiar property of gingiva is its rapid wound-healing process and the absence of scarring. Here we investigated the phenotype and the possible function of mesenchymal stem cells isolated from gingival papilla. Methods: MSCs have been isolated from gingival papilla (GinPa-MSCs) of donors undergoing oral surgery [1]. Primary cultures were analyzed for their proliferation rate, clonogenicity, mesenchymal stem cell marker expression [2] and multi-differentiative ability towards mesodermal lineages. Subsequently, we focused our research on the possible use of GinPa-MSCs as vehicles for drug delivery. Results: All the GinPa-MSCs showed a fibroblast-like morphology and a high proliferation rate (DT = 50.4±16.1 hours) that allowed us to rapidly collect a large number of cells. GinPa-MSCs displayed a high clonogenic potential (~20%) and expressed on their surface CD73, CD90, CD105 (>99%) and CD14 (~32%) while lacked CD45. CD14 mild surface expression and the abundance of pinocytotic structures, as revealed by TEM analysis, may be related to GinPa-MSC functional activity of active defence in the oral environment. When induced to differentiate towards osteogenic lineage, GinPa-MSCs increased collagen production (+79%) and extracellular calcified matrix deposition (+100%) respect to control cells, despite their high basal ALP activity. Preliminary data also indicated that GinPa-MSCs possess a mild chondrogenic potential, while lipid droplets, following adipogenic stimuli, were poorly detectable. After being tested for their sensitivity to the compound, GinPa-MSCs were primed with the chemotherapy drug Paclitaxel (PTX). Our results show that GinPa-MSCs do not metabolize or inactivate PTX and they can up-take and release it in an amount able to inhibit in vitro the growth of CFPAC-1 carcinoma cells. Conclusions: Our data indicate that GinPa-MSCs display peculiar characteristics that could be related to their unique functions of tissue regeneration and insult defence within the oral environment. Furthermore, their ability to uptake and release PTX represents an interesting perspective for drug delivery approaches to treat cancer and other pathologies related to the odontostomatological apparatus. Taken together, our results suggest that gingival stromal cells represent an attractive candidate in maxillo-facial and dental regenerative medicine.