Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.     

Age-related changes in the brain of the dog.

Although many age-related changes have been described in the nervous system of different species, few authors have specifically studied the topic. Knowledge of such changes is essential to veterinary pathologists, who must distinguish the lesions of specific pathologic processes from those arising as a result of normal aging. The brains of 20 old dogs, ranging in age from 8 to 18 years, were compared with those of 10 young dogs using routine staining techniques (hematoxilin and eosin, periodic acid-Schiff), special staining techniques (periodic acid-methenamine silver stain), and immunohistochemical techniques to detect glial fibrillary acid protein, neurofilaments, ubiquitin, and beta-amyloid. Changes affected meninges and choroid plexuses, meningeal and parenchymal vessels, neurons, and glial cells. Of special interest was the presence of polyglucosan bodies, cerebrovascular amyloid deposition, senile plaques, and ubiquitinated bodies. Some of the age-related changes found, particularly lipofuscin, polyglucosan bodies, and beta-amyloid protein deposition, may play a role in the pathogenesis of the canine cognitive dysfunction syndrome. The dog could be used as a natural animal model for the study of normal aging and human neurodegenerative diseases.     

Dynamics in serum of the inflammatory markers serum amyloid A (SAA), haptoglobin,fibrinogen and alpha2-globulins during induced noninfectious arthritis in the horse

Despite the importance of noninfectious joint diseases in equine medicine, little is known about the acute phase response which may be elicited if the local inflammatory process of noninfectious arthritis is sufficiently strong, Therefore the aim of this study was to monitor the systemic inflammatory response during experimentally-induced noninfectious arthritis by studying the dynamics in serum of the acute phase proteins serum amyloid A (SAA), haptoglobin, fibrinogen and alpha2-globulins. Twenty-four Standardbred horses, age 3-7 years, found healthy on thorough clinical, radiological, haematological and serum biochemical examination, were injected aseptically into the right midcarpal joint with amphotericin B. Blood samples were drawn before induction of arthritis (0 h), and at 8, 16, 24, 36 and 48 h postinduction and then on Days 3, 4, 5 and 15 postinduction. All horses developed lameness with joint effusion and joint heat as well as increased respiratory rate, heart rate and body temperature. The lameness started to decline after 24-36 h and, in most animals, systemic signs disappeared on Day 2 postinjection. The concentration of the acute phase proteins increased following induction of arthritis. The SAA concentrations were higher than baseline concentrations from 16 h postinduction and were maximal at 36-48 h (227 times baseline concentration). The haptoglobin concentrations were higher than baseline concentrations from 24 h and were maximal at 48-96 h (1.14 times baseline concentration). The maximal concentrations of fibrinogen were seen between 36-72 h postinjection and increased on average 0.87 times from baseline concentrations. The fibrinogen concentrations were higher than baseline concentrations from 24 h postinjection. Alpha2-globulins concentrations showed a minor increase and increased 0.55 times from baseline concentrations. The markers had returned to baseline concentrations by Day 15. Our results demonstrate that amphotericin B-induced arthritis in a single joint gives rise to a systemic acute phase response measurable as increased concentrations in serum SAA, haptoglobin, fibrinogen and alpha2-globulins during the first 2 weeks of the condition and, thereby, that such an increase need not be indicative of infectious arthritis. Further research should be aimed at determining whether chronic noninfectious arthritis in the horse gives rise to increased acute phase protein concentrations in serum.     

Reno-, hepato- and splenomegaly of common carp fingerlings (Cyprinus carpio L.) diseased in swimbladder inflammation caused by Sphaerospora renicola Dyková et Lom, 1982

The weight of internal organs (swimbladder, kidney, liver, spleen) in relation to the body weight was studied in common carp fingerlings divided into three groups on the basis of swimbladder appearance and microscopic examination of the kidney. The fish had been collected from different Hungarian fish farms at the time when swimbladder inflammation (SBI) usually occurs (in July and August). The first group comprised fish with severe signs of SBI and massive renal sphaerosporosis, the second group consisted of fish with milder swimbladder changes and/or kidney infection by a low number of Sphaerospora renicola, while the third group was constituted by infection-free common carp fry. Statistical analysis of swimbladder, kidney, liver and spleen weight in relation to the body weight revealed that in the infected groups the internal organs were substantially enlarged. This suggests that in common carp fry with SBI the swimbladder changes are accompanied by reno-, hepato- and splenomegaly.     

The surgical technique and the age of the horse both influence the outcome of mosaicplasty in a cadaver equine stifle model

Six pieces of grafts, 6.5 mm in diameter, 20 mm in length, were taken from each of 170 cadaver hindlimbs, using the cranial surface of the medial femoral trochlea for harvesting. The age of the horses varied between 4 months and 23 years. 30 limbs under the age of 12 years were selected for transplantation. Three of six grafts were transplanted into the medial femoral condyle using different combinations of tunnel depth and dilation. With ageing, a significant decline in transplantability was detected. In general, mosaicplasty cannot be recommended in horses above 11 years. Based on a previous clinical case (Bodó et al., 2000), a good surface alignment was indeed achieved with a combination of graft length drilling and dilation in most cases. However, the occasional entrapment of cartilage debris under the graft prevented perfect alignment in the present cadaver study in 27% of the grafts transplanted in this manner. Since the protrusion of grafts never exceeded 1.5 mm, we conclude that drilling 3-5 mm deeper than graft length with graft length deep dilation can avoid disadvantageous protrusion of the transplanted hyaline cartilage caps, achieving bone decompression at the same time.     

Epidemiological, clinical, echo-doppler characteristics of mitral valve endocardiosis in Cavalier King Charles in France: a retrospective study of 451 cases (1995 to 2003)

A study performed on 451 Cavalier King Charles showed that 40.6% of dogs had a left apical systolic heart murmur, whose prevalence increased with age (> 11-year-old, 100%), but was not different between males and females. Mitral valve endocardiosis represented 93.3% of the ultrasonographic abnormalities.     

The Effect of Supplementation with Fresh Browse of Ziziphus mauritiana or Combretum aculeatum on Feed Intake,Nitrogen Utilization and Growth of Grazing Range Sheep

The influence of 0, 60 or 120 min access to a grove of either Ziziphus mauritiana (Ziziphus) or Combretum aculeatum (Combretum) on forage intake and on the digestibility, growth and excretion of nutrients was determined using 40 Oudah rams grazing for 7 h/day on poor-quality dry season pasture in the Sahel. The effects of browse species and browsing duration (0, 30, 60 or 120 min) on the ruminal ammonia content were also evaluated using 8 mature fistulated rams. Ziziphus increased both total digestible organic matter and total dry matter intakes per (kg live weight)^0.75 without decreasing herbage intake, whereas 60 or 120 min access to Combretum reduced herbage intake by 6.3% and 4.2%, respectively. The digestibility of the diet decreased (p<0.05) with the duration of access to the groves. Despite the provision of more nitrogen (N) than in the control diet, ruminal NH₃-N decreased 24 h after browsing commenced. The reduced live weight gain of sheep browsing Combretum may indicate more deleterious compounds in Combretum than in Ziziphus. Ziziphus appears to have more potential than Combretum to increase sheep production in low-input crop/livestock systems.     

The epidemiology and diagnosis of bluetongue with particular reference to Corsica

Bluetongue (BT) and/or BT viruses (BTV) have been identified in the Mediterranean basin and the Balkans each year from 1998 to 2002 and in particular BTV serotype 2 in the French Island of Corsica (2000 and 2001). In response to these virus incursions, the French Veterinary Authorities carried out epidemiological studies that included virological, serological and entomological analysis, and two vaccination campaigns performed in the winter of 2000/2001 and the winter and spring of 2001 and 2002. Rapid and reliable serotype differentiation is essential at the start of an outbreak to allow an early selection of vaccine to control the spread of the virus. Thus, molecular tools, that complement conventional methods, have been developed for early detection of infection, determination of the serotype, and differentiation between natural infection and vaccination. Serological results showed that the first vaccination campaign during the winter of 2000/2001 did not provide full protection for all sheep and during the summer of 2001, 335 sheep flocks in Corsica were again infected by BTV 2 (7-fold more that in 2000). Entomological studies have demonstrated that the only proven vector of the disease, Culicoides imicola, was present in the island in 2000 and that it has successfully established itself in Corsica. The safety and immunogenicity of the commercial South African vaccine were studied. Fourteen sheep were vaccinated and then observed for clinical signs. Blood, sera, spleen and lymph nodes were collected and analyzed, and the results confirmed the safety and potency of using this vaccine to protect sheep from clinical disease. As a result, an intensive vaccination campaign was performed during winter and spring 2001/2002. No cases of BT had been observed by the end of summer 2002, indicating that the vaccination campaign has been successful in protecting sheep from infection.     

Evaluation of hematological,biochemical and oxidative stress profile in calves under propofol anesthesia

Veterinary Research Communications - Propofol is a widely used drug in veterinary medicine to induce anesthesia; as well as the chosen compound for protocols of intravenous anesthesia. The present...