A novel ventilated thermoplastic mesh bandage for post-operative management of large soft tissue defects: a case series of three dogs treated with autologous platelet concentrates

A ventilated thermoplastic mesh bandage was used for the post-operative management of large soft tissue defects in three dogs. Once the granulation tissue appeared, the wounds were treated with liquid or jellified autologous platelet concentrates, Platelet Rich Plasma (PRP) and Platelet Lysate (PL), to improve the wound healing process. After cleaning the wound with sterile physiological solution, a dressing was performed with several layers of cotton. A window through the layers of cotton was opened above the wound. Then, the platelet concentrate was topically applied, and the bandage was completed by placing, over the access window, a ventilated thermoplasticmeshmodeled according to the size and shape of the wound. After 24 h, it was replaced by a low adhesion bandage. The thermoplastic mesh avoids the direct contact between the wound and the external layers of the bandage, preventing the drainage of the topical agent and the removal of the growing healthy granulation tissue. The bandage proposed in this study is easily applied by the veterinarian and well-tolerated by the animal, ensuring high welfare standards in stressed patients presenting compromised clinical conditions

In Vitro Evaluation of Cytotoxicity and Proliferative Effects of Lyophilized Porcine Liver Tissue on HepG2 Hepatoma Cells and Adipose-Tissue-Derived Mesenchymal Stromal Cells

In recent years, nutritional supplements from different sources have been widely considered to support medical treatments in patients affected by chronic hepatopathies. Their potential therapeutic benefit has been recognized, but some evidence of safety issues has been reported. Recently it has been hypothesized that the liver could produce various of bioactive factors to maintain organ homeostasis and promote tissue healing. Thus, liver-specific preparations containing bioactive factors could provide a suitable substrate for in vitro study of liver tissue maintenance/healing, as a prospective regenerative medicine approach. Furthermore, they could represent a dietary supplement or nutraceutical for adjuvant therapies when correctly prepared and formulated. This work aims to provide data about the safety and biological activity of a freeze-dried porcine liver preparation. The lyophilized powder obtained from the whole organ has been tested in term of in vitro cell cytotoxicity (MTT assay) and proliferation assays (bromo-deoxyuridine incorporation and direct cell count) in two different cell types: human hepatoma HepG2 cell line and adipose-tissue-derived canine mesenchymal stromal cells (At-MSCs). At concentration levels between 100 to 500 g/mL, the lyophilized liver powder stimulated mitochondrial metabolism as assessed by MTT assay (p 0.001 for HepG2 and for At-MSCs) and induced an increase in bromo-deoxyuridine incorporation in both cell types (p 0.01 for HepG2 and p < 0.001 for At-MSCs). In addition, direct cell count demonstrated a higher proliferative activity in treated At-MSCs (p < 0.001). Although preliminary, these data suggest that the whole-liver powder is noncytotoxic in vitro and may represent a stimulus to cell metabolism and proliferation. Further studies are needed to detect the bioactive components of the supplement and characterize in deeper detail the cellular pathways that they can modulate

Redox Status in Canine Leishmaniasis

World Health Organization defined Leishmaniasis as ‘one of the priority attention diseases’. Aiming to clarify some aspects of its pathogenetic mechanisms, our study has been focused on the assessment of redox status in dogs, the main reservoir for Leishmania infantum. Forty-five dogs from an endemic area in southern Italy were divided into four different groups (from mild disease with negative to low positive antibody levels to very severe disease with medium to high positive antibody levels) accordingly to the LeishVet group guidelines. Their plasma and/or sera were tested for Reactive Oxygen Species (ROS), namely superoxide anion (O2-), Reactive Nitrogen Species (RNS), such as nitric oxide (NO) and hydroperoxides (ROOH), as well as the activity of the detoxifying enzyme superoxide dismutase (SOD), and the total non-enzymatic antioxidant capacity, as determined by ferric reducing-antioxidant power (FRAP) assay, O2- generation was significantly (p &lt; 0.05) reduced in leishmaniasis affected dogs independently of the clinical stage, while NO production was stimulated (p &lt; 0.05) only in II and III stage patients. No difference could be found for the levels of hydroperoxides and SOD activity between healthy and pathological subjects. FRAP values, were lower in affected dogs but only in stage II. Taken together, although we demonstrate that several redox status parameters are altered in the plasma of dog affected by Leishmaniasis, the oxidative stress changes that are observed in this disease, are possibly mainly due to the cellular blood components i.e. neutrophils, responsible of the elimination of the parasite. Further studies are required to assess the clinical values of the collected data

Characterization of a Deswapped Triple Mutant Bovine Odorant Binding Protein

The stability and functionality of GCC-bOBP, a monomeric triple mutant of bovine odorant binding protein, was investigated, in the presence of denaturant and in acidic pH conditions, by both protein and 1-aminoanthracene ligand fluorescence measurements, and compared to that of both bovine and porcine wild type homologues. Complete reversibility of unfolding was observed, though refolding was characterized by hysteresis. Molecular dynamics simulations, performed to detect possible structural changes of the monomeric scaffold related to the presence of the ligand, pointed out the stability of the β-barrel lipocalin scaffold

Evaluation of Oxidative Stress Parameters in Healthy Saddle Horses in Relation to Housing Conditions, Presence of Stereotypies, Age, Sex and Breed

Oxidative stress plays an important role in the development of many horse diseases and it has been shown that housing has important implications for the psychophysical well-being of horses. The aim of this study is to determine if there are any dierences between the redox status in horses in relation to housing conditions. The four housing conditions analyzed were: single box, without external access and without contact (Cat A), single box with external access and possibility of partial contact (Cat B), group housing with box and large paddock (Cat C), pasture with more than 7 horses and the possibility of green forage for the whole year (Cat D). A group of 117 healthy horses were selected in several private stables in Northern Italy. All subjects treated with any type of drug were excluded. At the end of the enrollment, the 117 selected horses were divided into the four housing categories. Stereotypies were highest in the group of horses in single box, without external access and without contact (Cat A). Oxidative stress was evaluated by testing plasma or serum samples for the following parameters: superoxide anion (WST), nitric oxide (NO), reactive oxygen species (d-ROMs), ferric reducing ability of plasma (FRAP), and the activity of superoxide dismutase (SOD). Simultaneously with the blood sampling, the owners completed a questionnaire with all the management aspects of the horse (signaling, feeding, equestrian activity, vaccinations, foot management etc.). The statistical evaluation was carried out based on the categories previously described, on the presence and absence of stereotypies and on some signaling data obtained from the questionnaire. There were no significant dierences in the parameters analyzed between the categories. No significant redox status dierences were detected based on the presence or absence of stereotypies. Interestingly, when the age was introduced as selection (&lt;14 and &gt;14 years old) parameter inside the categories, statistical significance was observed for some of the stress markers considered. Finally, independently of the housing conditions, the horses of the most two represented breeds exhibited dierent values of FRAP. All these aspects are commented in the discussion

Xenobiotic-Free Medium Guarantees Expansion of Adipose Tissue-Derived Canine Mesenchymal Stem Cells both in 3D Fibrin-Based Matrices and in 2D Plastic Surface Cultures

Mesenchymal stem cells (MSCs) have been recently introduced in veterinary medicine as a potential therapeutic tool for several pathologies. The large-scale in vitro expansion needed to ensure the preparation of a suitable number of MSCs for clinical application usually requires the use of xenogeneic supplements like the fetal bovine serum (FBS). The substitution of FBS with species-specific supplements would improve the safety of implanted cells, reducing the risk of undesired immune responses following cell therapy. We have evaluated the effectiveness of canine adipose tissue-derived stromal vascular fraction (SVF) and MSCs (ADMSCs) expansion in the presence of canine blood-derived supplements. Cells were cultured on traditional plastic surface and inside a 3D environment derived from the jellification of different blood-derived products, i.e., platelet-poor plasma (PPP), platelet-rich plasma (PRP), or platelet lysate (PL). PPP, PRP, and PL can contribute to canine ADMSCs in vitro expansion. Both allogeneic and autologous PPP and PL can replace FBS for ADMSCs culture on a plastic surface, exhibiting either a similar (PPP) or a more effective (PL) stimulus to cell replication. Furthermore, the 3D environment based on homospecific blood-derived products polymerization provides a strong stimulus to ADMSCs replication, producing a higher number of cells in comparison to the plastic surface environment. Allogeneic or autologous blood products behave similarly. The work suggests that canine ADMSCs can be expanded in the absence of xenogeneic supplements, thus increasing the safety of cellular preparations. Furthermore, the 3D fibrin-based matrices could represent a simple, readily available environments for effective in vitro expansion of ADMSCs using allogeneic or autologous blood-products

Immunophenotypical characterization of canine mesenchymal stem cells from perivisceral and subcutaneous adipose tissue by a species-specific panel of antibodies

Immunophenotypical characterization of mesenchymal stem cells is fundamental for the design and execution of sound experimental and clinical studies. The scarce availability of species-specific antibodies for canine antigens has hampered the immunophenotypical characterization of canine mesenchymal stem cells (MSC). The aim of this study was to select a panel of species-specific direct antibodies readily useful for canine mesenchymal stem cells characterization. They were isolated from perivisceral and subcutaneous adipose tissue samples collected during regular surgeries from 8 dogs. Single color flow cytometric analysis of mesenchymal stem cells (P3) deriving from subcutaneous and perivisceral adipose tissue with a panel of 7 direct anti-canine antibodies revealed two largely homogenous cell populations with a similar pattern: CD29+, CD44+, CD73+, CD90+, CD34−, CD45− and MHC-II− with no statistically significant differences among them. Antibody reactivity was demonstrated on canine peripheral blood mononuclear cells. The similarities are reinforced by their in vitro cell morphology, trilineage differentiation ability and RT-PCR analysis (CD90+, CD73+, CD105+, CD44+, CD13+, CD29+, Oct-4+ gene and CD31− and CD45− expression). Our results report for the first time a comparison between the immunophenotypic profile of canine MSC deriving from perivisceral and subcutaneous adipose tissue. The substantial equivalence between the two populations has practical implication on clinical applications, giving the opportunity to choose the source depending on the patient needs. The results contribute to routine characterization of MSC populations grown in vitro, a mandatory process for the definition of solid and reproducible laboratory and therapeutic procedures

Markers of neutrophil mediated inflammation associate with disturbed continuous electroencephalogram after out of hospital cardiac arrest

Background Achieving an acceptable neurological outcome in cardiac arrest survivors remains challenging. Ischemia-reperfusion injury induces inflammation, which may cause secondary neurological damage. We studied the association of ICU admission levels of inflammatory biomarkers with disturbed 48-hour continuous electroencephalogram (cEEG), and the association of the daily levels of these markers up to 72 h with poor 6-month neurological outcome. Methods This is an observational, post hoc sub-study of the COMACARE trial. We measured serum concentrations of procalcitonin (PCT), high-sensitivity C-reactive protein (hsCRP), osteopontin (OPN), myeloperoxidase (MPO), resistin, and proprotein convertase subtilisin/kexin type 9 (PCSK9) in 112 unconscious, mechanically ventilated ICU-treated adult OHCA survivors with initial shockable rhythm. We used grading of 48-hour cEEG monitoring as a measure for the severity of the early neurological disturbance. We defined 6-month cerebral performance category (CPC) 1-2 as good and CPC 3-5 as poor long-term neurological outcome. We compared the prognostic value of biomarkers for 6-month neurological outcome to neurofilament light (NFL) measured at 48 h. Results Higher OPN (p = .03), MPO (p < .01), and resistin (p = .01) concentrations at ICU admission were associated with poor grade 48-hour cEEG. Higher levels of ICU admission OPN (OR 3.18; 95% CI 1.25-8.11 per ln[ng/ml]) and MPO (OR 2.34; 95% CI 1.30-4.21) were independently associated with poor 48-hour cEEG in a multivariable logistic regression model. Poor 6-month neurological outcome was more common in the poor cEEG group (63% vs. 19% p < .001, respectively). We found a significant fixed effect of poor 6-month neurological outcome on concentrations of PCT (F = 7.7, p < .01), hsCRP (F = 4.0, p < .05), and OPN (F = 5.6, p < .05) measured daily from ICU admission to 72 h. However, the biomarkers did not have independent predictive value for poor 6-month outcome in a multivariable logistic regression model with 48-hour NFL. Conclusion Elevated ICU admission levels of OPN and MPO predicted disturbances in cEEG during the subsequent 48 h after cardiac arrest. Thus, they may provide early information about the risk of secondary neurological damage. However, the studied inflammatory markers had little value for long-term prognostication compared to 48-hour NFL.Peer reviewe

ECMO for COVID-19 patients in Europe and Israel

Since March 15th, 2020, 177 centres from Europe and Israel have joined the study, routinely reporting on the ECMO support they provide to COVID-19 patients. The mean annual number of cases treated with ECMO in the participating centres before the pandemic (2019) was 55. The number of COVID-19 patients has increased rapidly each week reaching 1531 treated patients as of September 14th. The greatest number of cases has been reported from France (n = 385), UK (n = 193), Germany (n = 176), Spain (n = 166), and Italy (n = 136) .The mean age of treated patients was 52.6 years (range 16–80), 79% were male. The ECMO configuration used was VV in 91% of cases, VA in 5% and other in 4%. The mean PaO2 before ECMO implantation was 65 mmHg. The mean duration of ECMO support thus far has been 18 days and the mean ICU length of stay of these patients was 33 days. As of the 14th September, overall 841 patients have been weaned from ECMO support, 601 died during ECMO support, 71 died after withdrawal of ECMO, 79 are still receiving ECMO support and for 10 patients status n.a. . Our preliminary data suggest that patients placed on ECMO with severe refractory respiratory or cardiac failure secondary to COVID-19 have a reasonable (55%) chance of survival. Further extensive data analysis is expected to provide invaluable information on the demographics, severity of illness, indications and different ECMO management strategies in these patients