Cytochrome P450 3A, NADPH cytochrome P450 reductase and cytochrome b5 in the upper airways in horse

Gene and protein expression as well as catalytic activity of cytochrome P450 (CYP) 3A were studied in the nasal olfactory and respiratory mucosa and the tracheal mucosa of the horse. We also examined the activity of NADPH cytochrome P450 reductase (NADPH P450 reductase), the amount of cytochrome b(5) and the total CYP content in these tissues. Comparative values for the above were obtained using liver as a control. The CYP3A related catalytic activity in the tissues of the upper airways was considerably higher than in the liver. The CYP3A gene and protein expression, on the other hand, was higher in the liver than in the upper airway tissues. Thus, the pattern of CYP3A metabolic activity does not correlate with the CYP3A gene and protein expression. Our results showed that the activity of NADPH P450 reductase and the level of cytochrome b(5) in the relation to the gene and protein expression of CYP3A were higher in the tissues of the upper airways than in the liver. It is concluded that CYP3A related metabolism in horse is not solely dependent on the expression of the enzyme but also on adequate levels of NADPH P450 reductase and cytochrome b(5).     

Non-invasive assessment of growth, gender and time of day related changes of respiratory pattern in healthy cats by use of barometric whole body plethysmography

The objective of this study was to establish a reference base for respiratory variables (respiratory rate [R(R)], inspiratory and expiratory time [T(i) and T(e)], peak inspiratory and expiratory pseudoflow [PIF and PEF], tidal volume [V(T)], minute ventilation [V(E)] and enhanced pause [Penh]) of healthy cats by use of barometric whole body plethysmography (BWBP). Eighteen healthy European cats (10 male, 8 female) were studied from the age of 3 to 13 months in order to assess growth- and gender-related changes of BWBP variables. Chest radiographs and bronchoalveolar lavage cytology were performed to confirm pulmonary health status. Diurnal changes were investigated every 2 h over a period of 24 h when the cats were adult. V(T), V(E), PIF and PEF significantly increased during somatic growth and were higher in males than in females, whereas R(R), T(i), T(e), T(e)/T(i) ratio, PEF/PIF ratio and Penh remained unchanged and were not affected by gender. When measured over 24 h, Penh, T(e) and T(i) were significantly increased in the early morning hours (04:00 h), whereas R(R), PIF and PEF were decreased at that time. This study provides reference values of BWBP variables for healthy male and female cats and indicates when circadian changes might be observed.     

Effects of boar presence on agonistic behavior, shoulder scratches, and stress response of bred sows at mixing

The objective of this study was to determine the effects of boar presence on aggression, shoulder scratches, and salivary cortisol in group-housed sows during the period after mixing, which is when dominance hierarchies are formed. A total of 225 York-shire sows were used. Five groups of 15 sows were each exposed to 1 of 3 levels of boar presence (n = 15): physical (boar in pen with sows; 2.15 m2 per sow), fenceline (boar housed adjacent to sows; 2.3 m2 per sow), or control (no boar in the room; 2.3 m2 per sow). The experiment was divided into 2 phases. In phase 1, behavioral measurements were taken for 48 h after mixing and boar introduction. In phase 2, behavioral measurements were taken beginning on d 6 after mixing and included 24 h before and 48 h after boar removal. In phases 1 and 2, shoulder scratches were scored 24 h before and 24 h after mixing and boar removal, respectively. Saliva samples were collected each morning (0600 to 0700) and afternoon (1600 to 1700) during both phases. Frequencies of intersow aggressive contact (bite, body knock, and head knock) and threats as well as frequencies and durations of fighting bouts were determined from video recordings during daily feeding (49.6 +/- 1.4 min) and nonfeeding (21 +/- 0.2 h) periods. During phase 1, boar presence did not affect the frequency of threats during either the feeding or nonfeeding periods. Control groups had fewer aggressive contacts at feeding during the 25- to 48-h period compared with the fenceline groups (P = 0.01). Total duration of fighting and average fighting bout duration were unaffected by treatment. However, the number of fights occurring during feeding 25 to 48 h postmixing was lower in the physical groups than in the fenceline groups (P = 0.023); measures for the control groups were intermediate. Physical sows also received fewer shoulder scratches postmixing than did control sows (P = 0.048). After mixing, physical sows showed greater morning (P = 0.08) and afternoon (6 h postmixing, P = 0.01; 32 h postmixing, P = 0.08) salivary cortisol concentrations compared with fenceline sows, although they were only numerically greater than those for control sows. During phase 2, removing the boar did not increase fighting among sows or affect shoulder scratches or salivary cortisol concentrations. The presence of a boar was minimally effective at reducing fighting and scratches during the postmixing period, and sows showed a greater stress response in the presence of a boar.     

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.     

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.     

ENDOSCOPIC ULTRASOUND INSTRUMENTATION, APPLICATIONS IN HUMANS, AND POTENTIAL VETERINARY APPLICATIONS

Endoluminal scanning under endoscopic guidance, or endoscopic ultrasonography (EUS), has become the most significant advance for imaging the gastrointestinal (GI) tract wall and contiguous organs in the past 20 years. It was originally designed to overcome the limitations in humans to imaging the abdominal organs transabdominally, such as large penetration depths and GI air. This imaging modality provides detailed images of pathological processes both within and outside of the GI wall since a high-frequency transducer can be brought into close proximity with the target regions. It has found most success in humans for the staging of lung, gastric, and esophageal cancer, the detection of both lymphatic and hepatic metastases, and diagnosis of pancreatitis and pancreatic cancer, as well as achieving an important role in interventional and therapeutic procedures. The EUS examination can be performed to examine both the thorax and abdomen in animals when both conventional transthoracic or transabdominal ultrasound are inadequate due to intervening air, bone, large penetration depths, or obesity. The echoendoscope is similar to a conventional endoscope but has an ultrasound transducer at its tip. Both radial and linear multifrequency scanners are available. Linear scanners allow fine-needle aspiration (FNA) of the bowel wall or extraluminal structures. Transducer coupling is either by direct mucosal contact or by inflation of a water-filled balloon surrounding the transducer. Current thoracic applications for EUS in veterinary medicine include examination of the mediastinum, bronchial lymph nodes, esophagus, and pulmonary lesions as well as FNA of pulmonary masses. Abdominal applications include examination of both pancreatic limbs and the liver, including portosystemic shunts, detection of lymphadenomegaly, and examination of the gastric wall, duodenum, and jejunum. Other potential applications in dogs and cats include tumor staging and intrapelvic ultrasound.     

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.