Dilated cardiomyopathy (DCM) is a myocardial disease of dogs and humans characterized by progressive ventricular dilation and depressed contractility and it is a frequent cause of heart failure. Conventional pharmacological therapy cannot reverse the progression of the disease and, in humans, cardiac transplantation remains the only option during the final stages of cardiac failure. Cytoprotective gene therapy with the Vascular Endothelial Growth Factor-B167 (VEGF-B167) has proved an effective alternative therapy, halting the progression of the disease in experimental studies on dogs [1,2]. The aim of this work was to test the tolerability and feasibility of intracoronary inoculation under fluoroscopic guidance of VEGF-B167 carried by adeno-associated viral vectors in canine DCM patients. Ten patients underwent the gene delivery procedure. The intraoperative phase was well tolerated by all dogs. Clinical and echocardiographic assessment at 7 days post-procedure in all dogs showed stable clinical conditions that could be superimposed to those pre-procedure. The results of this work indicate that intracoronary gene delivery is feasible and tolerated in dogs with DCM. Further monitoring/investigations are ongoing to evaluate the effects of this procedure on disease progre

Introduction: Intercellular communication governs the exchange of signalling molecules between cells. The different mechanisms employed in cell-to-cell communication include the secretion and receptor-mediated binding of diffusible messengers such as hormones and growth factors, the transport of small molecules through gap junctions, exosomes as well as tunnelling nanotubes. In this study, we report a new feature of intercellular transfer in cultured sheep bone marrow mesenchymal stem cells (BM-sMSCs). Methods: Bone marrow was harvested from iliac crest of healthy sheep. Mononuclear cells were isolated by gradient centrifugation, plated in culture flasks and incubated in the presence of Coon’s medium in a humidified atmosphere at 37 °C with 5% CO2. Non-adherent cells were discarded after 3 days and adherent cells were cultured until they reached near-confluence (10 days). The cells were trypsinized and the pellets were fixed with 3% glutaraldehyde and post-fixed in OsO4 1%. After dehydration and embedding in Epon 812, ultrathin sections were counterstained with lead citrate and uranyl acetate and observed at a transmission electron microscope. Results: The cell populations consisted of two undifferentiated cell types: electron-lucent and electron-dense cells. Both showed prominent RER, glycogen aggregates and filopodia. The electron-lucent cells were more numerous. The nucleus in electron-lucent cells was euchromatic and contained a prominent nucleolus, whereas in electron-dense cells it was irregular in shape and full of heterochromatin. No intercellular junctions were observed between either electron-lucent cells or electron-dense cells, whereas spot-like fusions of the plasma membrane occur between the electron-lucent cell surface and electron-dense cells. Across this distinct membranous connection, a flow of cytoplasm - but not of organules – occurs from electron-dense to electron-lucent cells. Conclusions: This finding shows a new cell-to-cell connection implicated in intercellular transfer and stimulates further studies to identify the cell types involved and the nature of the substances transferred.

The aim of this study was to evaluate the correlation of commonly used oxygenation indices with venous admixture (Qs/Qt) in anaesthetised horses under different infusion rates of dobutamine. Six female horses were anaesthetised with acepromazine, xylazine, diazepam, ketamine, and isoflurane, and then intubated and mechanically ventilated with 100% O2. A Swan-Ganz catheter was introduced into the left jugular vein and its tip advanced into the pulmonary artery. Horses received different standardised rates of dobutamine.For each horse, eight samples of arterial and mixed venous blood were simultaneously obtained at fixed times. Arterial and venous haemoglobin (Hb) concentration and O2 saturation, arterial oxygen partial pressure (PaO2), venous oxygen partial pressure (PvO2), and barometric pressure were measured. Arterial (CaO2), mixed venous (CvO2), and capillary (Cc'O2) oxygen contents were calculated using standard formulae. The correlations between F-shunt, arterial oxygen tension to fraction of inspired oxygen ratio (PaO2/FiO2), arterial to alveolar oxygen tension ratio (PaO2/PAO2), alveolar to arterial oxygen tension difference (P[A - a]O2), and respiratory index (P[A - a]O2/PaO2) were tested with linear regression analysis. The goodness-of-fit for each calculated formula was evaluated by means of the coefficient of determination (r2). The agreement between Qs/Qt and F-shunt was analysed with the Bland-Altman test.All tested oxygen tension-based indices were weakly correlated (r2 < 0.2) with the Qs/Qt, whereas F-shunt showed a stronger correlation (r2 = 0.73). F-shunt also showed substantial agreement with Qs/Qt independent of the dobutamine infusion rate. F-shunt better correlated with Qs/Qt than other oxygen indices in isoflurane-anaesthetised horses under different infusion rates of dobutamine

The aim of this study was to compare intravenous regional anesthesia (IVRA) and brachial plexus block (BPB) for intra-operative analgesia in dogs undergoing pancarpal arthrodesis (PA). Twenty dogs scheduled for PA were intramuscularly sedated with acepromazine (0.03mg/kg), general anesthesia was intravenously (IV) induced with thiopental (10mg/kg) and, after intubation, maintained with isoflurane in oxygen. In 10 dogs (GIVRA) IVRA was performed on the injured limb administering 0.6ml/kg of 0.5% lidocaine. In 10 dogs (GBPB) the BPB was performed at the axillary level with the help of a nerve stimulator and 0.3ml/kg of a 1:1 solution of 2% lidocaine and 1% ropivacaine was injected. During surgery fentanyl (0.002mg/kg IV) was administered if there was a 15% increase of HR and/or MAP compared to the values before surgical stimulation. All the standard cardiovascular and respiratory parameters were continuously monitored during surgery. The duration of surgery and the time of extubation were recorded. Data were compared with a 1-way ANOVA test (P<0.05). No patients required fentanyl administration during surgery. All the recorded parameters were similar in the two groups. The two techniques were similar in providing intra-operative analgesia in dogs undergoing orthopaedic surgery.

The aim of this study was to compare the postoperative analgesic effects of robenacoxib and buprenorphine alone or in combination, in cats after ovariohysterectomy. Thirty healthy cats were randomly assigned to receive buprenorphine (0.02 mg/kg, n = 10; GB), robenacoxib (2 mg/kg, n = 10; GR) or their combination at the same dosages (n = 10; GBR) SC. After 30 min cats were sedated with an IM administration of medetomidine (0.02 mg/kg) and ketamine (5 mg/kg). General anaesthesia was induced with propofol and after intubation was maintained with isoflurane. Before premedication and at 1, 2, 3, 4, 6, 8, 12 and 24 h after extubation, pain and sedation were assessed using a simple descriptive pain scale, ranging from 0 (no pain/no sedation) to 4 (intense pain/ deep sedation). If the pain score was ≥3, rescue analgesia was provided using buprenorphine (0.02 mg/kg) administered IM. Pain score was higher in GB at 2, 3, 4, 6 and 8 h compared to baseline and compared to GBR at the same study times. Moreover, the pain score was also higher in GB compared to GR at 2, 3, 4 and 6 h. Pain score was similar at all study times between GR and GBR. Sedation at 1 and 2 h was higher than baseline values in all groups. Cats in GB received rescue analgesia more often than cats assigned to GR or GBR. Robenacoxib was an effective analgesic drug in cats up to 24 h after ovariohysterectomy. The addition of buprenorphine did not provide any additional analgesic effects compared to robenacoxib alone. © 2013 Elsevier Ltd. All rights reserved.

Klotho is an anti-aging factor mainly produced by renal tubular epithelial cells (TEC) with pleiotropic functions. Klotho is down-regulated in acute kidney injury in native kidney; however, the modulation of Klotho in kidney transplantation has not been investigated. In a swine model of ischemia/reperfusion injury (IRI), we observed a remarkable reduction of renal Klotho by 24 h from IRI. Complement inhibition by C1-inhibitor preserved Klotho expression in vivo by abrogating nuclear factor kappa B (NF-kB) signaling. In accordance, complement anaphylotoxin C5a led to a significant down-regulation of Klotho in TEC in vitro that was NF-kB mediated. Analysis of Klotho in kidneys from cadaveric donors demonstrated a significant expression of Klotho in pre-implantation biopsies; however, patients affected by delayed graft function (DGF) showed a profound down-regulation of Klotho compared with patients with early graft function. Quantification of serum Klotho after 2 years from transplantation demonstrated significant lower levels in DGF patients. Our data demonstrated that complement might be pivotal in the down-regulation of Klotho in IRI leading to a permanent deficiency after years from transplantation. Considering the anti-senescence and anti-fibrotic effects of Klotho at renal levels, we hypothesize that this acquired deficiency of Klotho might contribute to DGF-associated chronic allograft dysfunction.

NADPH oxidase plays a central role in mediating oxidative stress during heart, liver, and lung ischemia/reperfusion injury, but limited information is available about NADPH oxidase in renal ischemia/reperfusion injury. Our aim was to investigate the activation of NADPH oxidase in a swine model of renal ischemia/reperfusion damage. We induced renal ischemia/reperfusion in 10 pigs, treating 5 of them with human recombinant C1 inhibitor, and we collected kidney biopsies before ischemia and 15, 30, and 60 min after reperfusion. Ischemia/reperfusion induced a significant increase in NADPH oxidase 4 (NOX-4) expression at the tubular level, an upregulation of NOX-2 expression in infiltrating monocytes and myeloid dendritic cells, and 8-oxo-7,8-dihydro-2'-deoxyguanosine synthesis along with a marked upregulation of NADPH-dependent superoxide generation. This burden of oxidative stress was associated with an increase in tubular and interstitial expression of the myofibroblast marker α-smooth muscle actin (α-SMA). Interestingly, NOX-4 and NOX-2 expression and the overall NADPH oxidase activity as well as α-SMA expression and 8-oxo-7,8-dihydro-2'-deoxyguanosine synthesis were strongly reduced in C1-inhibitor-treated animals. In vitro, when we incubated tubular cells with the anaphylotoxin C3a, we observed an enhanced NADPH oxidase activity and α-SMA protein expression, which were both abolished by NOX-4 silencing. In conclusion, our findings suggest that NADPH oxidase is activated during ischemia/reperfusion in a complement-dependent manner and may play a potential role in the pathogenesis of progressive renal damage in this setting. Copyright © 2014 Elsevier Inc. All rights reserved.

Aim of this study was to document the normal computed tomographic tenography findings of digital flexor tendon sheath. Six ex vivo normal equine forelimbs were used. An axial approach was used to inject 185 mg/mL of iopamidol in a total volume of 60 mL into the digital flexor tendon sheaths. Single-slice helical scans, with 5 mm thickness, spaced every 3 mm, for a pitch of 0.6, and with bone algorithm reconstruction, were performed before and after injections of contrast medium. To obtain better image quality for multiplanar reconstruction and 3D reformatting, postprocessing retroreconstruction was performed to reduce the images to submillimetre thickness. Computed tomographic tenography of digital flexor tendon sheaths could visualize the following main tendon structures for every forelimb in contrast-enhanced images as low densities surrounded by high densities: superficial digital flexor tendon, deep digital flexor tendon, manica flexoria, mesotendons, and synovial recess. Results of this study suggest that computed tomographic tenography can be used with accuracy and sensitivity to evaluate the common disorders of the equine digital flexor tendon sheath and the intrathecal structures.

Brucella spp. is a worldwide zoonotic pathogen. Infection by Brucella canis in dogs is endemic in the Southern USA and in Central and South America, but it appears sporadically in other parts of the world, including Europe. Tissue samples from a dog with chronic prostatitis, discospondylitis and locomotor problems were subjected to clinical and laboratory examinations. B. canis was detected by PCR in biological fluids and tissues of the animal, while antibodies to B. canis were found in the serum, providing additional strong evidence for the circulation of B. canis in Italy.

The aim of this study was to compare the cardiovascular effects of medetomidine, acepromazine and their combination administered intravenously in healthy dogs. Ten dogs were included in this study and randomly assigned to the three different sedative protocols: medetomidine (2 μg/kg, protocol M), acepromazine (20 μg/kg, protocol A) and acepromazine followed by medetomidine with the same doses as above (protocol AM), in three different times. In all subjects before (Tbase) and 15 (T15), 50 (T50) and 80 (T80) minutes after the administration of the drugs, the following non-invasive measurements were obtained: blood pressure with oscillometric method, ECG, and echocardiography. Blood pressure and echocardiography evidenced decrease in left ventricular afterload secondary to acepromazine and an increase in right ventricular afterload due to medetomidine. The combination of the two drugs mitigated the effects expected by the single drugs used alone, and prevented the onset of atrioventricular blocks, such as seen in protocol M. The three protocols were eligible for sedation and premedication in healthy dogs. Moreover they had little impact on the echocardiographic variables evaluated in this study.

To evaluate the effects of 10 cm H(2)O of positive end-expiratory pressure (PEEP) on lung aeration and gas exchange in mechanically ventilated sheep during general anesthesia induced and maintained with propofol. ANIMALS: 10 healthy adult Bergamasca sheep. PROCEDURES: Sheep were sedated with diazepam (0.4 mg/kg, IV). Anesthesia was induced with propofol (5 mg/kg, IV) and maintained with propofol via constant rate infusion (0.4 mg/kg/min). Muscular paralysis was induced by administration of vecuronium (25 microg/kg, bolus IV) to facilitate mechanical ventilation. After intubation, sheep were positioned in right lateral recumbency and mechanically ventilated with pure oxygen and zero end-expiratory pressure (ZEEP). After 60 minutes, 10 cm H(2)O of PEEP was applied for 20 minutes. Spiral computed tomography of the thorax was performed, and data were recorded for hemodynamic and gas exchange variables and indicators of respiratory mechanics after 15 (T(15)), 30 (T(30)), and 60 (T(60)) minutes of ZEEP and after 20 minutes of PEEP (T(PEEP)). Computed tomography images were analyzed to determine the extent of atelectasis before and after PEEP application. RESULTS: At T(PEEP), the volume of poorly aerated and atelectatic compartments was significantly smaller than at T(15), T(30), and T(60), which indicated that there was PEEP-induced alveolar recruitment and clearance of anesthesia-induced atelectasis. Arterial oxygenation and static respiratory system compliance were significantly improved by use of PEEP. CONCLUSIONS AND CLINICAL RELEVANCE: Pulmonary atelectasis can develop in anesthetized and mechanically ventilated sheep breathing pure oxygen; application of 10 cm H(2)O of PEEP significantly improved lung aeration and gas exchange.

OBJECTIVE: To evaluate the effectiveness of reduction of inspired oxygen fraction (Fio(2)) or application of positive end-expiratory pressure (PEEP) after an alveolar recruitment maneuver (ARM) in minimizing anesthesia-induced atelectasis in dogs. ANIMALS: 30 healthy female dogs. PROCEDURES: During anesthesia and neuromuscular blockade, dogs were mechanically ventilated under baseline conditions (tidal volume, 12 mL/kg; inspiratory-to-expiratory ratio, 1:2; Fio(2), 1; and zero end-expiratory pressure [ZEEP]). After 40 minutes, lungs were inflated (airway pressure, 40 cm H(2)O) for 20 seconds. Dogs were then exposed to baseline conditions (ZEEP100 group), baseline conditions with Fio(2) reduced to 0.4 (ZEEP40 group), or baseline conditions with PEEP at 5 cm H(2)O (PEEP100 group; 10 dogs/group). For each dog, arterial blood gas variables and respiratory system mechanics were evaluated and CT scans of the thorax were obtained before and at 5 (T5) and 30 (T30) minutes after the ARM. RESULTS: Compared with pre-ARM findings, atelectasis decreased and Pao(2):Fio(2) ratio increased at T5 in all groups. At T30, atelectasis and oxygenation returned to pre-ARM findings in the ZEEP100 group but remained similar to T5 findings in the other groups. At T5 and T30, lung static compliance in the PEEP100 group was higher than values in the other groups. CONCLUSIONS AND CLINICAL RELEVANCE: Application of airway pressure of 40 cm H(2)O for 20 seconds followed by Fio(2) reduction to 0.4 or ventilation with PEEP (5 cm H(2)O) was effective in diminishing anesthesia-induced atelectasis and maintaining lung function in dogs, compared with the effects of mechanical ventilation providing an Fio(2) of 1.

Objective To compare the effects of two fractions of inspired oxygen (FiO 2) (0.4 and 1) on lung aeration and gas exchange during general anaesthesia in cats.Study design Randomized, blinded, controlled study.Animals Thirty healthy, mixed breed, client owned female cats.Materials and methods Cats were premedicated intramuscularly with acepromazine (0.03 mg kg -1) and medetomidine (0.015 mg kg -1). Anaesthesia was induced with propofol (5 mg kg -1) and, after orotracheal intubation, maintained with isoflurane carried by either 100% oxygen (G100, n = 15) or an oxygen-air mixture with 40% oxygen (G40, n = 15). All cats were placed in dorsal recumbency and breathed spontaneously throughout the entire procedure. Following surgery (ovariectomy), a spiral computed tomography (CT) of the thorax was performed, arterial oxygen (PaO 2) and carbon dioxide (PaCO 2) tensions were measured and alveolar-arterial gradient of oxygen [P(A-a)O 2] calculated. The CT images were analysed for lung aeration by the analysis of radiograph attenuations (Hounsfield units, HU), according to the following classification: hyperinflated area (-1000 to -900 HU), normally aerated area (-900 to -500 HU), poorly aerated area (-500 to -100 HU) and non-aerated area (-100 to +100 HU). The groups were compared using one-way anova.Results Compared to G100, the normally-aerated lung area was significantly greater and the poorly-aerated and non-aerated areas were significantly smaller in G40. PaCO 2 was similar in both groups. PaO 2 and P(A-a)O 2 were significantly higher in G100. In both groups, pulmonary atelectasis developed preferentially in the caudal lung fields.Conclusion In cats anaesthetised with isoflurane, the administration of an FiO 2 of >0.9 significantly impaired lung aeration and gas exchange as compared to an FiO 2 of 0.4.Clinical relevance An FiO 2 of 0.4 may better preserve lung aeration and gas exchange in anaesthetised spontaneously breathing cats but monitoring is essential to ensure oxygenation is adequate

The pathophysiology of endotoxemia-induced acute kidney injury (AKI) is characterized by an intense activation of the host immune system and renal resident cells by lipopolysaccharide (LPS) and derived proinflammatory products. However, the occurrence of renal fibrosis in this setting has been poorly investigated. The aim of the present study was to investigate the possible association between endothelial dysfunction and acute development of tissue fibrosis in a swine model of LPS-induced AKI. Moreover, we studied the possible effects of coupled plasma filtration adsorption (CPFA) in this setting.

Background. Increasing evidence demonstrates a phenotypic plasticity of endothelial cells (ECs). Endothelial-to-mesenchymal transition (EndMT) contributes to the development of tissue fibrosis. However, the pathogenic factors and signalling pathways regulating this process in ischaemia/reperfusion (I/R) injury are still poorly understood. Methods. We investigated the possible role of complement in the induction of this endothelial dysfunction in a swine model of renal I/R injury by using recombinant C1 inhibitor in vivo. Results. Here, we showed that I/R injury reduced the density of renal peritubular capillaries and induced tissue fibrosis with generation of CD31+/α-SMA+ and CD31+/FPS-1+ cells indicating EndMT. When we inhibited complement, the process of EndMT became rare, with preserved density of peritubular capillaries and significant reduction in renal fibrosis. When we activated ECs by anaphylatoxins in vitro, C3a and C5a led to altered endothelial phenotype with increased expression of fibroblast markers and decrease expression of specific endothelial markers. The activation of Akt pathway was pivotal for the C3a and C5a-induced EndMT in vitro. In accordance, inhibition of complement in vivo led to the abrogation of Akt signalling, with hampered EndMT and tissue fibrosis.

Field immobilization of captive antelope may be required for medical examination, blood sample collection, and animal identification. The aim of this study was to evaluate the effects of a combination of butorphanol, detomidine, and midazolam (BDM) and its partial reversibility in Nile Lechwe antelope (NLA; Kobus megaceros). Nine captive NLAs, weighing 28-64 kg, were immobilized, in February 2011, with butorphanol 0.20±0.05 (meanSD) mg/kg, detomidine 0.20±0.05 mg/kg and midazolam 0.31±0.08 mg/kg administered intramuscularly (IM) with a blowpipe. Physiologic parameters and depth of anesthesia were recorded when the animals became recumbent at 19.55±8.36 min after darting (T0) and after 10 (T10), 20 (T20) and 30 (T30) min. An arterial blood sample was collected at T20. At the end of the procedures, immobilization was partially reversed with atipamezole 0.25 mg/kg IM. Quality of induction, immobilization, and recovery were scored. The BDM combination induced immobilization and lateral recumbency in 13.44±5.61 min. Median induction score [scored 1 (excellent) to 4 (poor)] was 1 (range 1–2). Heart rate varied 40–104 beats/min, respiratory rate 16–108 breaths/min and rectal temperature 36.5–40.3 C. Hyperthermia observed and rapidly treated in three animals that demonstrated insufficient immobilization after darting. Arterial blood gas analyses revealed a mean pH of 7.43±0.07, PaCO2 of 44.1±6.0 mmHg, PaO2 of 74.0±13.5 and a SaO2 of 94.77±3.96 mmHg. Recovery was smooth and animals were walking in 13.44±7.85 min. Median recovery score [scored 1 (excellent) to 4 (poor)] was 1 (range 1–2). The BDM was effective in immobilizing captive healthy NLAs with minimal cardiorespiratory changes.

The aims of this study were to evaluate the effects of the administration of a combination of tiletamine-zolazepam and detomidine (TZD) in 9 tigers (Panthera tigris). Nine captive tigers were immobilized with tiletamine-zolazepam and detomidine administered intramuscularly. At the end of the procedure immobilization was partially reversed with atipamezole. Lateral recumbency was achieved in 15.6±5.9min. The median induction score [scored 1 (excellent) to 4 (poor)] was 1. The immobilization score [scored 1 (poor) to 6 (too deep)] was 5 (4-5) at all study times. After atipamezole administration, all tigers experienced severe ataxia and incoordination. Median recovery score [scored 1 (excellent) to 4 (poor)] was 2.5 (range 2-3). No neurologic and/or important adverse reactions were noticed within 5 days after recovery. The combination tiletamine-zolazepam with detomidine proved to be effective in immobilizing captive healthy tigers but it maybe associated with hypertension and ataxia during recovery.

The aim of this study was to evaluate the influence of abdominal surgery on atelectasis formation in healthy dogs. After the induction of general anesthesia (GA), 20 dogs, scheduled for elective ovariohysterectomy, were positioned in dorsal recumbency: 10 dogs underwent immediate surgery (S group), while 10 dogs (NS group) were maintained under anesthesia for 60 min before surgery. In both groups, a helical computed tomography (CT) scan of the thorax and an arterial blood gas analysis were performed 60 min after the induction of GA. Lung aeration was estimated by analyzing the radiographic attenuation of the CT images. The atelectasic and poorly aerated lung compartments were significantly larger, and the normally aerated lung compartment was smaller in the S group compared to the NS group. The PaO2 was similar in both groups. Abdominal surgery significantly increases pulmonary atelectasis in healthy dogs under GA

The aim of this study was to evaluate the influence of abdominal surgery on atelectasis formation in healthy dogs. After the induction of general anesthesia (GA), 20 dogs, scheduled for elective ovariohysterectomy, were positioned in dorsal recumbency: 10 dogs underwent immediate surgery (S group), while 10 dogs (NS group) were maintained under anesthesia for 60 min before surgery. In both groups, a helical computed tomography (CT) scan of the thorax and an arterial blood gas analysis were performed 60 min after the induction of GA. Lung aeration was estimated by analyzing the radiographic attenuation of the CT images. The atelectasic and poorly aerated lung compartments were significantly larger, and the normally aerated lung compartment was smaller in the S group compared to the NS group. The PaO2 was similar in both groups. Abdominal surgery significantly increases pulmonary atelectasis in healthy dogs under GA.

The aim of this study was to evaluate the influence of a recruiting maneuver (RM) on the effects of PEEP on lung function in healthy horses under general anaesthesia. Fifteen horses were sedated with acepromazine (0.02 mg kg-1 IV) and detomidine (0.005 mg kg-1 IV), general anaesthesia was intravenously induced with midazolam (0.1 mg kg-1) and ketamine (2.2 mg kg-1) and maintained with isoflurane in 100% oxygen. After intubation all horses lungs were mechanically ventilated in a volume controlled mode: Vt (12 ml kg-1) and I:E (1:2) were unchanged during the study while RR was titrated to maintain the PE’CO2 between 40 and 45 mmHg. Three different ventilatory strategies were applied in all horses during the same anaesthetic episode: zero PEEP (ZEEP), 10 cmH2O of PEEP (PEEP) and a RM followed by the application of 10 cmH2O of PEEP (RMPEEP). The RM was performed applying 50 cmH2O for 20 seconds to the respiratory system. Thirty minutes after each ventilatory strategy was initiated, HR and MAP were recorded, an arterial blood sample was collected [PaO2, P(A-a)O2], static compliance of the respiratory system (CRSstat) and the PEEP recruited lung volume (RLV) (Grasso S et al 2005) were calculated. Data were compared with the ANOVA test (P<0.05). The PaO2 and Crsstat were higher while P(A-a)O2 lower at RMPEEP (60.7 ± 6.8 kPa, 473.5 ± 89.1 ml cmH2O-1 and 23.8 ± 7.5 kPa) compared to ZEEP (40.8 ± 16.9 kPa, 339.7 ± 81.9 ml cmH2O-1 and 44.5 ± 16.8 kPa) and PEEP (41.2 ± 17.8 mmHg, 360.9 ± 54.9 ml cmH2O-1 and 44.4 ± 17.6 mmHg). The RLV was larger at RMPEEP (5.5 ± 2.4 L) than at PEEP (2.6 ± 1.6 L). A RM significantly improved the effects of 10 cmH2O of PEEP on lung function in horses under general anaesthesia.

This report details a bubble echocardiographic study carried out during the surgical treatment of a congenital single extrahepatic portosystemic shunt (PSS) in a Labrador Retriever. After celiotomy, agitated saline was injected through a jejunal vein and microbubbles appeared rapidly in the right cardiac chambers. The test confirmed the presence of a PSS, helping the surgeon to identify the vessel concerned and to rule out a second shunt. Successively, portography confirmed what the exploratory celiotomy had revealed before with the aid of the bubble study: a single shunt was located between the portal vein and the right renal vein. It was completely ligated, as all the criteria for this solution were met. Intraoperative contrast echocardiography (ICE) was easy to perform, helpful and undemanding. It is proposed here as an intraoperative ancillary test to diagnose all PSS and to confirm successful treatment when complete shunt closure is possible.

Objective: The Acute Respiratory Distress Syndrome Network protocol recommends limiting tidal volume and plateau pressure; it also recommends increasing respiratory rate to prevent hypercapnia. We tested a strategy that combines the low tidal volume with lower respiratory rates and minimally invasive CO2 removal. Subjects: Ten lung-damaged pigs (instilled hydrochloride). Interventions: Two conditions randomly applied in a crossover fashion: the Acute Respiratory Distress Syndrome Network protocol and the Acute Respiratory Distress Syndrome Network protocol plus lower respiratory rate plus minimally invasive Co2 removal. A similar arterial Co2 partial pressure was targeted in the two conditions. Measurements and Main Results: Physiological parameters, computed tomography scans, plasma and bronchoalveolar lavage concentrations of interleukin-1[beta], interleukin-6, interleukin-8, interleukin-10, interleukin-18, and tumor necrosis factor-[alpha]. During the lower respiratory rate condition, respiratory rate was reduced from 30.5 +/- 3.8 to 14.2 +/- 3.5 (p < 0.01) breaths/min and minute ventilation from 10.4 +/- 1.6 to 4.9 +/- 1.7 L/min (p < 0.01). The extracorporeal device removed 38.9% +/- 6.1% (79.9 +/- 18.4 mL/min) of CO2 production. During the lower respiratory rate condition, interleukin-6, interleukin-8, and tumor necrosis factor-[alpha] concentrations were significantly lower in plasma; interleukin-6 and tumor necrosis factor-[alpha] concentrations were lower in bronchoalveolar lavage, whereas the concentrations of the other cytokines remained unchanged. Conclusion: The strategy of lower respiratory rate plus minimally invasive extracorporeal CO2 removal was feasible and safe and, as compared with the Acute Respiratory Distress Syndrome Network protocol, reduced the concentrations of some, but not all, of the tested cytokines without affecting respiratory mechanics, gas exchange, and hemodynamics.

LPS-induced sepsis is a leading cause of acute kidney injury (AKI) in critically ill patients. LPS may induce CD80 expression in podocytes with subsequent onset of proteinuria, a risk factor for progressive chronic kidney disease (CKD) frequently observed after AKI. This study aimed to investigate the therapeutic efficacy of LPS removal in decreasing albuminuria through the reduction of podocyte CD80 expression. Between January 2015 and December 2017, 70 consecutive patients with Gram-negative sepsis-induced AKI were randomised to either have Coupled Plasma Filtration and Adsorption (CPFA) added to the standard care (n=35) or not (n=35). To elucidate the possible relationship between LPS-induced renal damage, proteinuria and CD80 expression in Gram- sepsis, a swine model of LPS-induced AKI was set up. 3-hours after LPS infusion, animals were treated or not with CPFA for 6-hours. Treatment with CPFA significantly reduced serum cytokines, CRP, procalcitonin and endotoxin levels in patients with Gram-negative sepsis-induced AKI. CPFA significantly lowered also proteinuria and CD80 urinary excretion. In the swine model of LPS-induced AKI, CD80 glomerular expression, which was undetectable in control pigs, was markedly increased at the podocyte level in LPS-exposed animals. CPFA significantly reduced LPS-induced proteinuria and podocyte CD80 expression in septic pigs. Our data indicate that LPS induces albuminuria via podocyte expression of CD80 and suggest a possible role of timely LPS removal in preventing the maladaptive repair of the podocytes and the consequent increased risk of CKD in sepsis-induced AKI.

Current treatment of leukemia focuses on increasing chemotherapy efficacy. Mesenchymal stromal cells (MSCs) have been proposed for carrying and delivery drugs to improve cancer cells-specific killing. Recently, we have shown that MSCs loaded with the anti-cancer drug Paclitaxel (PTX), acquire a potent anti-tumor activity. To expand these findings, we here investigate the effect of human MSCs (hMSCs) and mouse SR4987 loaded with PTX (hMSCsPTX and SR4987PTX) on Molt-4 and L1210, two Leukemia cell (LC) lines of human and mouse origin respectively. We found that SR4987PTX and hMSCsPTX had a strong anti-LC activity. HMSCsPTX co-injected with Molt-4 or intra-tumor injected into an established subcute Molt-4 nodules strongly inhibited growth and angiogenesis. In syngenic BDF1 mice-bearing L1210, the intraperitoneal administration of SR4987PTX doubled mice survival time. In vitro, both hMSCs and hMSCsPTX released chemotactic factors, bound and formed rosette with LCs. By ultrastructural analysis of rosette, hMSCsPTX showed no morphological alterations while the attached LCs were apoptotic and necrotic. Finally, MSCs and MSCsPTX released molecules that reduced LCs adhesion to microvascular endothelium (hMECs) and down-modulated the adhesion molecules ICAM-1 and VCAM-1 on hMECs. We conclude that, besides conventional chemotherapy, MSCsPTX could be a new therapeutic approach to treat Leukemia in humans.

Abstract Objective – To evaluate the feasibility and efficacy of noninvasive continuous positive airway pressure (CPAP) administered with a pediatric helmet in healthy dogs recovering from general anesthesia. Design – Randomized, cross-over, clinical study. Setting – University teaching hospital Animals – Fifteen healthy female, client-owned dogs recovering from general anesthesia following elective ovariohysterectomy. Interventions – All dogs received the same standardized anesthetic protocol (acepromazine, morphine, propofol, and isoflurane in oxygen). After extubation, a pediatric helmet was placed on all dogs and connected to a venturi valve supplied with medical air. In all patients, the gas flow was set to 50 L/minute and the FiO2 to 0.21. Dogs received the following sequence of treatments, each lasting 20 minutes: 0 CPAP (pre-CPAP), CPAP of 5 cm H2O (CPAP), and again 0 CPAP (post-CPAP). Measurements and Main Result – During the entire study, the following data were collected: pressure and FiO2 inside the helmet, mean arterial pressure, respiratory rate, heart rate, sedation score (0 = awake, 10 = deep sedation), and tolerance to the helmet (0 = excellent, 4 = poor). At the end of each phase, an arterial blood sample was sampled. As compared with the pre-CPAP and the post-CPAP periods, during the CPAP period, the PaCO2, alveolar-arterial oxygen gradient (P[A−a]O2), and respiratory rate significantly decreased. The PaO2 was higher at CPAP (105.6±4.0mmHg) compared with pre-CPAP (80.6±6.9mmHg) and post-CPAP (86.7 ± 5.8 mm Hg). Tolerance and sedation scores during the CPAP period were not different from those in the pre-CPAP and post-CPAP periods. Conclusions – Noninvasive CPAP applied through a helmet is a feasible and effective supportive technique in dogs recovering from general anesthesia.

The aim of this study was to evaluate the use of a single-portal endoscopic desmotomy of the PAL with the use of Arthrex - Centerline™. Ten horse distal front limbs from horses free of PAL disease were prepared for tenoscopy of the digital flexor tendon sheath (DFTS). A dual-port endoscopic desmotomy with a hook knife was performed in 5 specimens (Group A) and single port Arthrex-Centerline™ Desmotomy was performed in another 5 specimens (Group B). The performing time, judgments of the surgeon, number of blade passages, and gross anatomy were evaluated. The performing time and surgeon judgments were significantly lower in Group B. No significant differences were assessed in number of passages to achieve a complete PAL resection and a gross anatomy evaluation. The use of Arthrex-Centerline™ is feasible for a PAL desmotomy procedure. It was faster with more handling ease compared with the free-hand double-portal desmotomy and allowed the same results in terms of number of passages to complete the release evaluated at gross anatomy.

RATIONALE: In the presence of increased chest wall elastance, the airway pressure does not reflect the lung-distending (transpulmonary) pressure. OBJECTIVE: To compare the physiological effects of a conventional open lung approach titrated for an end-inspiratory airway opening plateau pressure (30 cm H2O) with a transpulmonary open lung approach titrated for a elastance-derived end-inspiratory plateau transpulmonary pressure (26 cm H2O), in a pig model of acute respiratory distress syndrome (HCl inhalation) and reversible chest wall mechanical impairment (chest wall and abdomen restriction). METHODS: In eight pigs, physiological parameters and computed tomography were recorded under three conditions: 1) conventional open lung approach, normal chest wall; 2) conventional open lung approach, stiff chest wall; and 3) transpulmonary open lung approach, stiff chest wall. MEASUREMENTS AND MAIN RESULTS: As compared with the normal chest wall condition, at end-expiration non aerated lung tissue weight was increased by 116 ± 68 % during the conventional open lung approach and by 28 ± 41 % during the transpulmonary open lung approach (p < .01), whereas cardiac output was decreased by 27 ± 19 % and 22 ± 14 %, respectively (p = not significant). CONCLUSION: In this model, the end-inspiratory transpulmonary open lung approach minimized the impact of chest wall stiffening on alveolar recruitment without causing hemodynamic impairment.

The purpose of this case series was to describe the historical, clinical, radiographic details, outcome and histhopathological findings of 2 cases of NSH in pony. CASE 1 A 3 years old, male pony was referred in 2003 for severe dyspnoea and deformity of the bones of the skull. At clinical examination, the pony had general fatigue, severe dyspnea and inspiratory breath sounds. X‐ray examination showed signs of severe thinning of the cranial bones with loss of definition of the plot medullary and cortical thinning cancellous bone. The results of parathyroid hormone by radioimmunological showed an increase in the values of PTH .Treatment has been provided using tiludronate (TILDREN 1mg/kg‐ ®‐Ceva Vetem SpA, Milan). The patient's clinical condition showed significant improvements, the clinical symptoms (inability to maintain the station, lameness and respiratory sounds) had resolved. The pony was dismissed. CASE 2 In October of 2012 was referred a 5 years old Shetland male pony, and the owners permanent recumbency since 3 days.General clinical examination showed persistent recumbency , slight enlargement of facial bones and pain at flexion of hindlimbs. Based on the clinical symptoms, X‐rays and laboratory tests, the diagnosis was of NSH related to fibrous osteodystrophy. The clinical condition of the horse showed no improvement and the subject was not able to take quadrupedal station. The owner disagrees with Tiludronate therapy and pony was euthanized, and some bone samples were harvested in order to deepen the diagnostic picture. Microradioghaphs of frontal serial sections show modifications of bone tissue arrangement and mineralization.. The trabecular network appears rarefied owing to a thinning of the trabeculae or their total erosion. Both spongy and compact bone frequently exhibit large osteocytic lacunae. On the other hand new bone, less mineralized, is exthensively present. The deposition fronts show a wide osteoid with voluminous osteoblast, but sometimes exhibit numerous preosteoblasts with a fibroblasts morphology. In many area, fibrous connective tissue covers the trabecular surfaces and replaces part of the marrow reticular stroma. The appearance of micro X‐ray, never described in the horse, and the histological evidence and confirm the diagnosis of fibrous osteodystrophy. Bisphosphonate therapy is a therapeutic option, and it is valid for the possible remission of clinical symptoms but not certain deformities of the disease. The association of such therapy with a balanced diet can be part of the treatment protocol in the course of NSH of dietary origin. David et all., the bisposphonate Tiludronate is a potent inhibitor of the osteoclast vacuola hatpase, Journal of Bone and Mineral Research, 11, 1498‐1507, 1996 Estepa, J. C., Aguilera‐Tejero, E., Mayer‐Valor, R., Almeden, Y., Felsenfeld, A.J. & Rodriguez, M. Measured of parathyroid hormone in horses. Equine Veterinary Journal 30, 476‐481, 1998

The veterinary literature has been enriched by numerous clinical studies and about the study of hip dysplasia in dogs, but there aren’t many works related to the feline species, although it is believed that it has a also considered important impact of increased life expectancy and changing eating and living conditions of cats. In this study, in order to diagnose hip dysplasia, the dorsolateral subluxation index of the femoral head in cats was determined by CT scans, simulating the loading position by comparing the data obtained with the angle of Norberg - Olsson. On the basis of analysis of the obtained data, it was found that the evaluation of the hip joint by this method can be predictive of the disease before the degenerative joint disease onset and with greater sensitivity than the Norberg angle measurement

The use of bone marrow-derived mesenchymal stem cells (MSCs) for clinical and experimental studies is increasing, but full characterization of MSCs in veterinary species is hindered by the variability in species-specific cell surface marker expression and antibody cross reactivity. Recent studies demonstrated that the glycans in the glycocalyx of MSCs are promising candidates as cell biomarkers. In the present study, we analysed the glycocalyx of canine MSCs (cMSCs), ovine MSCs (oMSCs), and equine MSCs (eMSCs) by using a cell microarray procedure in which MSCs were spotted on microarray slides and incubated with a panel of 14 biotinylated lectins and Cy3-conjugated streptavidin. The signal intensity was then detected by using a microarray scanner. The lectin-binding signals indicated that the MSC surface of the investigated species contained both N- and O-linked glycan types, with N-glycosylation predominating over O-glycosylation and fucosylation being more abundant than sialylation. Relative quantification revealed an interspecific difference between these glycans. In addition, cMSCs expressed more α2,3-linked sialic acid (MAL II), terminal lactosamine (RCA120), and α1,6 and α1,3 fucosylated oligosaccharides (PSA, LTA); oMSCs exhibited more T antigen (Jacalin), GalNAcα1,3(LFucα1,2)Galβ1,3/4GlcNAcβ1 (DBA), chitotriose (succinylated WGA), and α1,2-linked fucose (UEA I); and eMSCs showed a higher density of α2,6 sialic acids (SNA) and high mannose N-glycans (Con A). By using cell microarray methodology, we have for the first time demonstrated differences in the glycosylation profiles of cMSC, oMSC, and eMSC surfaces. These results could be valuable as resources and references for MSC differentiation and molecular remodeling in clinical cell-based therapy and tissue engineering studies.

Objective: The aim of this study was to track the survival and efficacy of allogeneic bone marrow mesenchymal stem cells (BM-MSC) marked with red fluorescent protein (BM-MSCRFP) in an ovine model of collagenaseinduced tendinopathy. Methods: Bone marrow was harvested from one donor sheep and BM-MSC were isolated, cultivated and transfected with red fluorescent protein (BM-MSCRFP). Collagenase was injected into both Achilles tendons in the remaining nine sheep. After two weeks the left tendon was injected with a solution of 6 × 106 BM-MSCRFP and fibrin glue, while only fibrin glue was administered to the contralateral tendon in each sheep. After three, four and six weeks the tendons were harvested and evaluated for morphology, collagen I deposition, presence of CD34+ cells, and fluorescent labelled BM-MSC. Results: We demonstrated that delivery of BM-MSC into tendon lesions had positive effects on the injured tendons. The BM-MSCRFP survived at three, four and six weeks after treatment, leading to better quality healing of tendons as compared to the controls, where no labelled cells were detected. Interestingly, we demonstrated high expression of CD34+ cells in tendons that had been treated with BM-MSCRFP. Clinical relevance: Mesenchymal stem cell allografts have a positive effect on tendon healing and local injection of BM-MSC directly into the tendon allows the homing of BM-MSC for good efficiency of engraftment.

Autologous bone marrow stromal cells (BMSC) implant after swine experimental myocardial infarct (MI) was investigated by serial technetium-99m (Tc-99m)-tetrofosmin gated single photon emission tomography (G-SPET) and compared with immuno-histochemical findings. The aim was to evaluate if intramyocardial BMSC implant produces any prolonged effect in the left ventricle (LV) perfusion and function. Eleven pigs underwent left anterior descending artery (LAD) ligature; in seven of them BMSC were injected in the border zone of the MI, while in the remaining four saline solution was injected at the same site. After LAD ligature G-SPET scans at 48h and at 5 and 10 weeks (w) after the implant were performed. Uptake defect size and LV function analysis were performed comparing 48h to 5w and 10w studies. Statistical evaluation was performed with Friedman test and unpaired Wilcoxon test. The comparison between a progressive reduction of Perfusion Image Score was observed from 48h to 5w and to 10w in the treated group (Friedman test: chi(2)= 13.56; P=0.01). No variation was observed in the control group (Friedman test: chi(2)=3; P= 0.223). Comparison of the absolute variation (A) between treated and control group resulted significant (Wilcoxon test W=10; P=0.007). Similar positive results were also observed for the relative extension of the uptake defect, wall motion and LVEF analysis. Histological data of our swine model demonstrated that autologous BMSC implanted in the damaged myocardium area had survived and differentiated into cells with typical features of myocardiocytes. Gated SPET is a reliable tool to evaluate prolonged positive effects of autologous BMSC implant in swine experimental MI model. In conclusion, autologous BMSC implanted can improve perfusion, induce cell regeneration, reduce wall motion abnormalities and prevent severe LV dysfunction in swines.

In the presence of risk factors, the anastomosis in the small intestine and in the colon are at risk for dehiscence and peritonitis. The apposition of a biological patch around the anastomosis might improve wound healing and therefore might prevent harmful, potentially life-threatening and costy complications. Aim: to verify if Tutomesh® facilitates the functional recovery of the intestinal anastomotic wound area (mucosa) in the pig ileum and colon. Methods: 24 Large White pigs (B.W. 25 kg; age 4-5 months) underwent ileal and colonic anastomosis with or without application of Tutomesh® and compared with healthy (intact) control intestinal segments. At days 2, 7, 14, 30 and 90 following surgery, ileal and colonic mucosa were isolated from similar anastomized and control tracts and mounted in Ussing chambers containing Krebs oxygenated solution at pH 7.4. Electrophysiological parameters, i.e. short circuit current (Isc) and transepithelial resistance (Rt), as markers of mucosal function, were continously measured by a digital voltage clamp system. Results: Ileal mucosa from control showed Isc of -17.10±4.72 μA/cm2 and Rt of 105.91±11.98 Ohm*cm2. In anastomized ileum Isc decreased by 52% and Rt increased by 58% (n=6 tissues); with Tutomesh® the Isc reduction was only 16.3% while Rt increased by 46% (for both n=6; p&lt;0.001 vs. control). In colonic mucosa from 13 control tissues, Isc was -10.67±2.29 μA/ cm2, Rt 140.94±14.38 Ohm*cm2. Colonic Isc and Rt (n=6) remained stable with anastom- osis, while Tutomesh® significantly increased the current by 47.0% (n=7; p&lt;0.001 vs. control). Conclusions: Our observations suggest that transport properties of intestinal mucosa improve significantly with Tutomesh® , a useful resorbable bio-patch which therefore helps the functional recovery of anastomoses, mainly in the ileum. Further studies are ongoing to assess the translational value of Tutomesh® in surgical patients.

Ischemia-reperfusion injury is the major cause of delayed graft function in transplanted kidneys, an early event significantly affecting long-term graft function and survival. Several studies in rodents suggest that the alternative pathway of the complement system plays a pivotal role in renal ischemia-reperfusion injury. However, limited information is currently available from humans and larger animals. Here we demonstrated that 30 minutes of ischemia resulted in the induction of C4d/C1q, C4d/MLB, and MBL/MASP-2 deposits in a swine model of ischemia-reperfusion injury. The infusion of C1-inhibitor led to a significant reduction in peritubular capillary and glomerular C4d and C5b-9 deposition. Moreover, complement-inhibiting treatment significantly reduced the numbers of infiltrating CD163(+), SWC3a(+), CD4a(+), and CD8a(+) cells. C1-inhibitor administration led to significant inhibition of tubular damage and tubular epithelial cells apoptosis. Interestingly, we report that focal C4d-deposition colocalizes with C1q and MBL at the peritubular and glomerular capillary levels also in patients with delayed graft function. In conclusion, we demonstrated the activation and a pathogenic role of classical and lectin pathways of complement in a swine model of ischemia-reperfusion-induced renal damage. Therefore, inhibition of these two pathways might represent a novel therapeutic approach in the prevention of delayed graft function in kidney transplant recipients.

Bone marrow mesenchymal stem cells (BM-MSCs) in combination with bioceramics have been used to repair bone lesions but the effects of these biomaterials on BM-MSCs are little known. This study reports the changes observed in the ultrastructure of sheep BM-MSCs incubated with silicon-stabilized tricalcium phosphate (TSP). MSCs were isolated from bone marrow of iliac crest and cultured according to Crovace et al. (VCOT, 21, 2008), TSP was added to some cultures and cells were incubated for 10 days. Control and TPS-incubated cells were fixed with GTA, post-fixed in OsO4 and embedded in Epon 812. Control and TSP cultures consisted of differently electron-dense polygonal cells containìng euchromatic nuclei with prominent nucleoli, In control coltures, the electron-lucent cells were characterized by dilated rER cisternae whereas the moderately electron-dense cells showed dark bodies as prominent features, Small blebs and filopodia were present on the surface of light and dark cells, respectively. In TSP cultures, thè light cells showed large surface blebs, peripheral cytoplasm poor in organelles which were packed with contractile filaments around the nucleus, The dark elongated cells displayed pseudopodia and filopodia, peripheral vacuoles, dense bodies, round or elongated mitochondria, These findings demonstrate that TSP modifies differently the ultrastructure of celi types constituting the BM-MSCs of sheep.

Bioceramics are being used in experimental bone engineering application in association with bone marrow derived mesenchymal stem cells (BM-MSCs) as a new therapeutic tool, but their effects on the ultrastructure of BM-MSCs are yet unknown. In this study we report the morphological features of ovine (o)BM-MSCs cultured with Skelite, a resorbable bioceramic based on silicon stabilized tricalcium phosphate (SiTCP), able to promote the repair of induced bone defect in sheep model. oBM-MSCs were isolated from the iliac crest, cultured until they reached near-confluence and incubated with SiTCP. After 48 hours the monolayers were highly damaged and only few cells adhered to the plastic. Thus, SiTCP was removed, and after washing the cells were cultured until they became confluent. Then, they were trypsinizated and processed for transmission electron microscopy (TEM) and RT-PCR analysis. RT-PCR displayed that oBM-MSCs express typical surface marker for mesenchymal stem cells. TEM revealed the presence of electron-lucent cells and electron-dense cells, both expressing the CD90 surface antigen. The prominent feature of electron-lucent cells was the concentration of cytoplasmic organelles around the nucleus as well as large surface blebs containing glycogen or profiles of endoplasmic reticulum. The dark cells had a multilocular appearance by the presence of peripheral vacuoles. Some dark cells contained endocytic vesicles, lysosomes, and glycogen aggregates. oBM-MSCs showed different types of specialized interconnections. The comparison with ultrastructural features of untreated oBM-MSCs suggests the light and dark cells are two distinct cell types which were differently affected by SiTCP bioceramic.

Ovine bone marrow-derived mesenchymal stromal cells (oBM-MSCs) represent a good animal model for cell-based therapy and tissue engineering. Despite their use as a new therapeutic tool for several clinical applications, the morphological features of oBM-MSCs are yet unknown. Therefore, in this study the ultrastructural phenotype of these cells was analysed by transmission electron microscopy (TEM). The oBM-MSCs were isolated from the iliac crest and cultured until they reached near-confluence. After trypsinization, they were processed to investigate their ultrastructural features as well as specific surface marker proteins by flow cytometry and immunogold electron microscopy. Flow cytometry displayed that all oBM-MSCs lacked expression of CD31, CD34, CD45, HLA-DR whereas they expressed CD44, CD58, HLAI and a minor subset of the cell population (12%) exhibited CD90. TEM revealed the presence of two morphologically distinct cell types: cuboidal electron-lucent cells and spindle-shaped electron-dense cells, both expressing the CD90 antigen. Most of the electron-lucent cells showed glycogen aggregates, dilated cisternae of RER, moderately developed Golgi complex, and secretory activity. The electron-dense cell type was constituted by two different cell-populations: type A cells with numerous endosomes, dense bodies, rod-shaped mitochondria and filopodia; type B cells with elongated mitochondria, thin pseudopodia and cytoplasmic connectivity with electron-lucent cells. These morphological findings could provide a useful support to identify “in situ” the cellular components involved in the cell-therapy when cultured oBM-MSCs are injected.

Objective-To evaluate the use of the oxygen content-based index, Fshunt, as an indicator of venous admixture (Qs/Qt) at various fractions of inspired oxygen (FIO2s) in anesthetized sheep undergoing 1-lung or 2-lung ventilation. Animals-6 healthy adult female sheep. Procedures-Sheep were anesthetized and administered 5 different FIO2s (0.21, 0.40, 0.60, 0.80, and 1.00) in random order during 2-lung mechanical ventilation. Arterial and mixed venous blood samples were obtained at each FIO2 after a 15-minute stabilization period. Vital capacity alveolar recruitment maneuvers were performed after blood collection. The previously used FIO2 sequence was reversed for sample collection during 1-lung ventilation. Blood samples were analyzed for arterial, pulmonary end-capillary, and mixed venous oxygen content and partial pressure and for hemoglobin concentration. Oxygen hemoglobin saturation, Qs/Qt, Fshunt, and oxygen tension-based indices (OTIs; including PaO2:FIO2, alveolar-arterial difference in partial pressure of oxygen [PAO2 -PaO2], [PAO2 -PaO2]:FIO2, [PAO2 -PaO2]:PaO2, and PaO2:PAO2) were calculated at each FIO2; associations were evaluated with linear regression analysis, concordance, and correlation tests. Intermethod agreement between Qs/Qt and Fshunt was tested via Bland-Altman analysis. Results-Strong and significant associations and substantial agreement were detected between Fshunt and Qs/Qt. Relationships between OTIs and Qs/Qt varied, but overall correlations were weak. Conclusions and Clinical Relevance-Whereas OTIs were generally poor indicators of Qs/Qt, Fshunt was a good indicator of Qs/Qt at various FIO2s, regardless of the magnitude of Qs/Qt, and could be potentially used as a surrogate for Qs/Qt measurements in healthy sheep.

The present invention concerns a process for attracting, recruiting and selecting mesenchymal cells with high bone tropism and marked bone regenerative ability, which can be used in the treatment of bone lesions of traumatic and pathological origin, in particular in cases of bone neoplasia.