José M. Gonzalo-Orden

Endogenous cardiac stem cell activation by insulin-like growth factor-1/hepatocyte growth factor intracoronary injection fosters survival and regeneration of the infarcted pig heart.

OBJECTIVES The purpose of this study was to test the ability of insulin-like growth factor (IGF)-1/hepatocyte growth factor (HGF) to activate resident endogenous porcine cardiac stem/progenitor cells (epCSCs) and to promote myocardial repair through a clinically applicable intracoronary injection protocol in a pig model of myocardial infarction (MI) relevant to human disease. BACKGROUND In rodents, cardiac stem/progenitor cell (CSC) transplantation as well as in situ activation through intramyocardial injection of specific growth factors has been shown to result in myocardial regeneration after acute myocardial infarction (AMI). METHODS Acute MI was induced in pigs by a 60-min percutaneous transluminal coronary angiography left anterior descending artery occlusion. The IGF-1 and HGF were co-administered through the infarct-related artery in a single dose (ranging from 0.5 to 2 μg HGF and 2 to 8 μg IGF-1) 30 min after coronary reperfusion. Pigs were sacrificed 21 days later for dose-response relationship evaluation by immunohistopathology or 2 months later for cardiac function evaluation by cardiac magnetic resonance imaging. RESULTS The IGF-1/HGF activated c-kit positive-CD45 negative epCSCs and increased their myogenic differentiation in vitro. The IGF-1/HGF, in a dose-dependent manner, improved cardiomyocyte survival, and reduced fibrosis and cardiomyocyte reactive hypertrophy. It significantly increased c-kit positive-CD45 negative epCSC number and fostered the generation of new myocardium (myocytes and microvasculature) in infarcted and peri-infarct/border regions at 21 and 60 days after AMI. The IGF-1/HGF reduced infarct size and improved left ventricular function at 2 months after AMI. CONCLUSIONS In an animal model of AMI relevant to the human disease, intracoronary administration of IGF-1/HGF is a practical and effective strategy to reduce pathological cardiac remodeling, induce myocardial regeneration, and improve ventricular function.

Effects of dynamic resistance training on fascicle length and isometric strength

Abstract The aims of this study were to assess changes in muscle architecture, isometric and dynamic strength of the leg extensor muscles, resulting from dynamic resistance training, and the relationships between strength and muscle architecture variables. The participants (n = 30) were randomly assigned to one of two groups. The training group (n = 16; age 21.8 ± 2.3 years, body mass 74.8 ± 9.2 kg, height 1.75 ± 0.08 m) performed dynamic resistance training for 13 weeks. The control group (n = 14; age 19.9 ± 1.5 years, body mass 74.0 ± 8.5 kg, height 1.76 ± 0.05 m) did not perform any resistance training. Maximal dynamic and isometric strength were tested in both groups, before and after the training period. The members of the training group used the free-weight squat lift (90°) as their training exercise. The concentric phase of the squat was performed explosively. Skeletal muscle architecture of the vastus lateralis was visualized using ultrasonography. At the end of the study, significant increases in vastus lateralis muscle thickness (+6.9%, P < 0.001), fascicle length (+10.3%, P < 0.05), one-repetition maximum (+8.2%, P < 0.05), rate of force development (+23.8%, P < 0.05) and average force produced in the first 500 ms (+11.7%, P < 0.05) were seen only in the training group. Adaptations to the muscle architecture in the training group limited the loss of fibre force, and improved the capacity for developing higher velocities of contraction. The architectural changes in the training group were similar to those seen in studies where high-speed training was performed. In conclusion, dynamic resistance training with light loads leads to increases in muscle thickness and fascicle length, which might be related to a more efficient transmission of fibre force to the tendon.

Diagnosis, genetic control and preventive management of canine hip dysplasia: a review.

Despite expensive screening and breeding programmes, hip dysplasia (HD) continues to be one of the most common orthopaedic diseases in dogs. The vast majority of dogs afflicted with HD show minimal to no clinical signs, but it can also be a highly debilitating condition for both working and pet dogs. Hip joint laxity is considered a major risk factor for the development of degenerative joint disease and a definitive diagnosis is made if characteristic signs are evident on a ventrodorsal view of the pelvis. Early prediction of the condition can be made using stress radiographic techniques to evaluate the passive hip laxity. The diagnosis of HD may be used for the purpose of selecting breeding stock or to decide on the best treatment approach. Breeding programmes based on individual dog phenotypes have been ineffective and a selection procedure based on breeding value (BV) estimation is recommended. Traditional conservative and surgical treatment approaches are reserved for dogs with overt clinical signs of the disease but such treatments can be expensive and aggressive, and are often ineffective in eliminating clinical signs or subluxation and in preventing the development of degenerative joint disease. The implementation of breeding programmes based on BVs and further research into early prediction/diagnosis of HD and effective preventive treatment approaches are essential.

Prevalence study of the lungworm Aelurostrongylus abstrusus in stray cats of Portugal

In this study we have investigated the prevalence of aelurostrongylosis, one of the most common feline pulmonary parasitic diseases, in cats from the north-west region of Portugal. For this purpose, 97 faecal samples were collected from cats at risk of being infected by Aelurostrongylus abstrusus in an animal shelter and in a municipal facility. Using the Baermann–Wetzel coprological technique, faecal shedding of first stage larvae (L1) was detected in 17.4% of the cats. Based on this result, it can be concluded that this lungworm infection seems to be common among feral cats in the north-west region of Portugal, in spite of the fact that clinical aelurostrongylosis is not frequently diagnosed by feline practitioners in the area. This parasitic disease should be included in the differential diagnosis of cats presenting with coughing or dyspnoea, and it also should be extended to asymptomatic animals with pulmonary nodules detected by image diagnosis.

Transient global ischemia in rat brain promotes different NMDA receptor regulation depending on the brain structure studied.

The mRNA expression of the major subunits of N-methyl-d-aspartate receptors (NR1, NR2A and NR2B) following ischemia-reperfusion was studied in structures with different vulnerabilities to ischemic insult in the rat brain. The study was performed using quantitative real-time PCR on samples from 3-month-old male Sprague-Dawley rats after global transient forebrain ischemia followed by 48h of reperfusion. Expression of NMDA receptor subunits mRNAs decreased significantly in all structures studied in the injured animals as compared to the sham-operated ones. The hippocampal subfields (CA1, CA3 and dentate gyrus) as well as the caudate-putamen, both reported to be highly ischemic-vulnerable structures, showed outstandingly lower mRNA levels of NMDA receptor subunits than the cerebral cortex, which is considered a more ischemic-resistant structure. The ratios of the mRNA levels of the different subunits were analyzed as a measure of the NMDA receptor expression pattern for each structure studied. Hippocampal areas showed changes in NMDA receptor expression after the insult, with significant decreases in the NR2A with respect to the NR1 and NR2B subunits. Thus, the NR1:NR2A:NR2B (1:1:2) ratios observed in the sham-operated animals became (2:1:4) in insulted animals. This modified expression pattern was similar in CA1, CA3 and the dentate gyrus, in spite of the different vulnerabilities reported for these hippocampal areas. In contrast, no significant differences in the expression pattern were observed in the caudate-putamen or cerebral cortex on comparing the sham-operated animals with the ischemia-reperfused rats. Our results support the notion that the regulation of NMDA receptor gene expression is dependent on the brain structure rather than on the higher or lower vulnerability of the area studied.

Coating titanium implants with bioglass and with hydroxyapatite: A comparative study in sheep

Summary.This study compares the osteointegration of titanium implants coated with bioglass (Biovetro GSB formula) and with hydroxyapatite (HAP). Twenty-four bioglass-coated and 24 HAP-coated cylinders were implanted in the femoral diaphyses of sheep, and examined after 2, 4, 6, 8, 12, and 16 weeks. The HAP coating gave a stronger and earlier fixation to the bone than did bioglass. Bioglass formed a tissue interface which showed a macrophage reaction with little new bone formation activity. In contrast, HAP, showed intense new bone formation, with highly mineralised osseous trabeculae in the neighbourhood of the interface.Résumé.Le but de cette étude expérimentale est de comparer l’ostéointégration d’implants en titane recouverts de bioverre (Biovetro GSD) et ceux recouverts d’hydroxyapatite (HAP). 24 cylindres recouverts de bioverre et 24 recouverts d’hydroxyapatite ont été implantés dans la diaphyse fémoral du mouton, puis examinés à 2, 4, 6, 8, 12 et 16 semaines de l’implantation. Les implants recouverts d’hydroxyapatite montrent une fixation à l’os plus précoce et plus solide que ceux recouverts de bioverre. Le bioverre a étéà l’origine d’un tissu d’interface os-implant avec présence d’une réaction macrophagique avec petite activité de formation d’os nouveau. L’hydroxyapatite en revanche, est à l’origine d’une formation assez intense d’os nouveau avec des trabécules bien minéralisés au voisinage de l’interface os-implant.

Global ischemia-induced modifications in the expression of AMPA receptors and inflammation in rat brain

Alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors (AMPAR) and inflammatory processes have been related to ischemia-induced damage, but there are few studies addressing their response in different brain areas. Here we compare AMPAR expression after ischemia in several brain areas (hippocampus, cerebral cortex and caudate-putamen) in an attempt to correlate it with their different vulnerabilities. We found outstanding decreases in GluR1 and GluR2 mRNA levels after global ischemia and 48 h reperfusion (I/R) in all the areas studied, however, protein levels maintained in some areas such as CA3, suggesting different post-transcriptional control in different areas of the brain. To characterize the inflammatory response in these areas, we measured the mRNA levels of CD11b/CD18 membrane integrin (a reactive microglia marker), which showed an important but similar up-regulation in all brain areas studied, which was confirmed by immunohistochemistry. We conclude that the down-regulation of AMPAR gene expression following I/R does not explain differences in the vulnerability of different areas. Additionally, our data indicate that the level of inflammation is independent of the vulnerability of the different brain areas and does not explain differences in the AMPAR expression observed in the brain areas studied.

Rolipram promotes functional recovery after contusive thoracic spinal cord injury in rats.

Numerous animal model studies in the past decade have demonstrated that pharmacological elevation of cyclic AMP (cAMP) alone, or in combination with other treatments, can promote axonal regeneration after spinal cord injury. Elevation of cAMP via the phosphodiesterase 4 (PDE4) inhibitor, rolipram, decreases neuronal sensitivity to myelin inhibitors, increases growth potential and is neuroprotective. Roliprams ability to cross the blood-brain barrier makes it a practical and promising treatment for CNS regeneration. However, several studies have questioned the efficacy of rolipram when given alone. The purpose of this investigation was to determine the effects of continuous administration of rolipram, given alone for 2 weeks, following a moderate T10 contusion injury in rat. Functional recovery was evaluated using the 21-point Basso, Beattie and Bresnahan (BBB) locomotor recovery scale and the beam walk. We used three-dimensional (3D) instrumented gait analysis to allow detailed assessment and quantification of hindlimb motion. The amount of the damaged tissue and spared white matter was estimated stereologically. Our results show that administration of rolipram following acute spinal cord contusion results in improved motor performance at each time-point. Dynamic assessment of foot motion during treadmill walking revealed a significantly decreased external rotation during the entire step cycle after 8 weeks in rolipram-treated animals. Stereological analysis revealed no significant differences in lesion volume and length. By contrast, spared white matter was significantly higher in the group treated with rolipram. Our results suggest a therapeutic role for rolipram delivered alone following acute SCI.

PASSIVE HIP LAXITY IN ESTRELA MOUNTAIN DOG - DISTRACTION INDEX, HERITABILITY AND BREEDING VALUES

Two hundred and fifteen Estrela Mountain Dogs (EMD) were examined using the PennHIP method between 2002 and 2006. Passive hip laxity (PHL) was estimated calculating the distraction index (DI). Pedigree information was obtained from the Portuguese Kennel Club. The heritability and breeding values were estimated using the linear Animal Model. The DI, using as reference the worst joint of each animal, ranged from 0.15 to 1.12 (0.55 +/- 0.19). The PHL was equal in right and left sides, and was higher in females than in males (P > 0.05 and P <0.01 in the t-test, respectively). The estimated heritability was 0.83 +/- 0.11. The mean breeding values for PHL were stable in EMD born between 1991 and 2003, and showed an improvement in 2004 and 2005. The data confirm high PHL in breeds with high prevalence and severity of canine hip dysplasia. The high heritability indicates that the DI could be reduced in the breed if PHL were used as a selection criterion. The early favourable evolution of DI breeding values could be related with the increase of voluntary radiographic hip screening in the last years, and the subsequent introduction of hip quality as a breeding criterion.

Pleural omentalisation with en bloc ligation of the thoracic duct and pericardiectomy for idiopathic chylothorax in nine dogs and four cats.

Conventional treatment of idiopathic chylothorax (IC) involves thoracic duct (TD) ligation (with/without lymphagiography) combined with subphrenic pericardiectomy. Nine dogs and four cats with IC, which received intrathoracic omentalisation with TD en bloc ligation (not preceded by lymphangiography) and subphrenic pericardiectomy, were evaluated retrospectively. Seven of nine dogs and 3/4 cats were still alive and disease-free at the time of reporting (range 10-53 and 19-31 months, respectively). Clinical signs of IC did not decrease after the first surgery in one cat and one dog; in another dog clinical signs recurred after 5 months. Overall efficacy rate of this one-stage combined procedure was 77% (6 months), 73% (12 months), and 57% (24 months). Where a second surgery was performed in case of failure, the success rate in dogs was 89% (6 months) and 80% (24 months). Addition of pleural omentalisation to TD en bloc ligation and subphrenic pericardiectomy does not seem to improve results when compared with published data and at present does not seem advisable as a first choice.