Severe myositis associated with Sarcocystis spp. infection in 2 dogs.

BACKGROUND: Dogs are definitive hosts for numerous species of the intracellular protozoan parasite Sarcocystis. Reports of sarcocysts in muscles of dogs most often represent incidental findings. HYPOTHESIS/OBJECTIVES: To report the clinicopathologic, ultrastructural, and molecular findings in 2 dogs with myositis associated with Sarcocystis spp. infection, as well as the response to treatment with antiprotozoal drugs. ANIMALS: Two dogs with severe myositis in association with massive sarcocystosis. METHODS: Retrospective case review. Affected dogs were identified by a diagnostic laboratory. Attending clinicians were contacted, and the medical records reviewed. Immunostaining and electron microscopy were performed on muscle biopsies. Biopsies also were subjected to 18S rRNA gene PCR. RESULTS: Both dogs had fever, lymphopenia, thrombocytopenia, and increased serum alanine aminotransferase (ALT) activity when first evaluated. One dog developed hyperbilirubinemia. Subsequently, both dogs had increased serum creatine kinase activity and clinical signs of myositis, with reluctance to move, generalized pain, and muscle wasting. Histopathology of muscle biopsies showed severe inflammatory and necrotizing myopathy with numerous sarcocysts. Ultrastructural studies and 18S rRNA gene sequence results were consistent with infection with a Sarcocystis spp. other than Sarcocystis neurona. Both dogs initially were treated unsuccessfully with clindamycin and anti-inflammatory drugs. One dog died. The other dog subsequently responded to treatment with decoquinate. CONCLUSIONS AND CLINICAL IMPORTANCE: Sarcocystis spp. infection should be included in the differential diagnosis for dogs that develop fever, thrombocytopenia, increased liver enzyme activities, and clinical and biochemical evidence of myositis. Although additional studies are required, decoquinate holds promise as an effective treatment for the disease.

Ion transport proteins and aquaporin water channels in the kidney of amphibians from different habitats.

Amphibians are known to spend part of their life on land and return to water to reproduce. However, some urodeles spend their entire life in water, while others succeed in completely avoiding water even during reproduction. Osmoregulatory mechanisms must therefore be different in the diverse environmental conditions of their respective life histories. The architecture of the kidney is similar in all amphibians; as a consequence the ion-water equilibrium must be regulated in the different environmental conditions. We investigated the immunolocalisation of Na(+)/K(+)/Cl(-) cotransport proteins, sodium pump and water-channel proteins (aquaporins) in aquatic Amphiuma means means, Rana dalmatina, a species that returns to water to reproduce, and Speleomantes genei, a completely terrestrial species. The investigation was carried out with immunohistochemical methods using antibodies to Na(+)/K(+)/Cl(-) cotransport protein NKCC1 T4, Na(+)/K(+)ATPase alpha-subunit, water-channel aquaporin 3 and the inner mitochondrial membrane (AMA). Cotransport proteins and sodium pump, involved in ion reabsorption, are widely distributed in A. means and R. dalmatina and confined to the distal segment in S. genei; conversely water channels, involved in water reabsorption, are limited to the collecting duct in A. means and R. dalmatina and distributed in the proximal and collecting ducts in S. genei.

Endothelial targeting with C1-inhibitor reduces complement activation in vitro and during ex vivo reperfusion of pig liver.

Tissue damage during cold storage and reperfusion remains a major obstacle to wider use of transplantation. Vascular endothelial cells and complement activation are thought to be involved in the inflammatory reactions following reperfusion, so endothelial targeting of complement inhibitors is of great interest. Using an in vitro model of human umbilical vein endothelial cells (HUVEC) cold storage and an animal model of ex vivo liver reperfusion after cold ischaemia, we assessed the effect of C1-INH on cell functions and liver damage. We found that in vitro C1-INH bound to HUVEC in a manner depending on the duration of cold storage. Cell-bound C1-INH was functionally active since retained the ability to inhibit exogenous C1s. To assess the ability of cell-bound C1-INH to prevent complement activation during organ reperfusion, we added C1-INH to the preservation solution in an animal model of extracorporeal liver reperfusion. Ex vivo liver reperfusion after 8 h of cold ischaemia resulted in plasma C3 activation and reduction of total serum haemolytic activity, and at tissue level deposition of C3 associated with variable level of inflammatory cell infiltration and tissue damage. These findings were reduced when livers were stored in preservation solution containing C1-INH. Immunohistochemical analysis of C1-INH-treated livers showed immunoreactivity localized on the sinusoidal pole of the liver trabeculae, linked to sinusoidal endothelium, so it is likely that the protective effect was due to C1-INH retained by the livers. These results suggest that adding C1-INH to the preservation solution may be useful to reduce complement activation and tissue injury during the reperfusion of an ischaemic liver.

Mitochondria-rich cells in gills and skin of an African lungfish, Protopterus annectens.

We used scanning electron microscopy, the vital dye DASPEI and an antibody to the inner mitochondrial membrane to study the presence and localisation of mitochondria-rich cells in the gills and skin (opercular, dorsal and ventral) of the lungfish Protopterus annectens in its free-swimming conditions and at the beginning of aestivation. In the free-swimming period, the gills were short and thick and the pavement cells were extremely large (30-40 microns). The mitochondria-rich cells, which were distributed in the secondary and primary epithelium, occurred as two morphologically different types, i.e. elongated and oval, similar to the alpha and beta chloride cells of fresh water teleosts. In the skin, only one type of mitochondria-rich cells was found, resembling the alpha chloride cells. All the mitochondria-rich cells distributed in the gills and skin were labelled with anti Ca(2+)-ATPase serum indicating the possible uptake of Ca2+ at freshwater chloride cell level. At the start of aestivation, the skin and gills were covered by a thick layer of mucus and the epithelium of the gills was reduced. The mitochondria-rich cells were almost completely covered by the pavement cells.

Simulated microgravity induces alteration in the central nervous system.

To investigate whether the signs of neurophysiological impairment observed in flight may be traced back to cytomorphology, we undertook a ground-based study focusing upon the architecture of cultured glial cells under simulated microgravity obtained by three-dimensional clinorotation.

Biotechnologies and acute hepatic failure.

Orthotopic liver transplantation survival for patients with acute liver failure is poor (50%). Mortality on the waiting list is high due to the lack of donors. For these reasons, the possibility of sustaining hepatic function by extra-corporeal liver perfusion must be considered. In this experimental research, two groups of pigs have been submitted to total de-vascularisation of the liver causing acute hepatic failure. In the first group (4 pigs) no extra-corporeal assistance has been used after total de-vascularisation. All pigs died between 16 and 33 hours after the acute hepatic failure was induced. In the second group (8 pigs) after complete hepatic de-vascularisation an extra-corporeal hepatic support by continuous allo-perfusion of isolated liver was performed using the Abouna-Costa extra-corporeal circuit. All pigs were observed during the acute hepatic failure which lasted from 6.30 to 7.30 hours. The data that were more positively influenced by the extra-corporeal assistance were ammonia and lactates that improved after the application of hepatic assistance.

Efficacy of intraoperative heat administration by ventilation with warm humidified gases and an oesophageal warming system.

We measured changes in body temperature in 12 hypothermic (mean aural temperature 34.4 (SD 1.0) degrees C) pigs during general anaesthesia with an open abdominal cavity and the effect of two warming systems: heating of inspired gases to 39 degrees C (intratracheal temperature) and oesophageal warming to 39 degrees C by a water perfused oesophageal heat exchanger. Each animal underwent both treatments and the control period in random sequence. Each condition was studied over 1 h. No additional protection against heat loss (drapes, blankets, i.v. fluids warming, etc.) was used. Anaesthesia, room temperature and relative humidity, amount and temperature of infusions and extension of exposed visceral surfaces were standardized. Mean decrease in body temperature was 1.0 (0.7) degree C (P < 0.005) without warming and 0.6 (0.2) degree C (P < 0.005) with heated inspired gases: this difference was not statistically significant. Oesophageal warming was very efficient as mean body temperature did not change significantly (-0.1 (0.2) degree C; ns).

Long-term stability of clinical attachment following guided tissue regeneration and conventional therapy.

UNLABELLED: 44 patients (34% smokers) presenting with severe periodontitis were treated with full mouth root planing (RPL). In each patient, 1 intrabony defect was treated with guided tissue regeneration (GTR). After 1 year of monthly prophylaxis, full mouth plaque (FMPS) and bleeding (FMBS) scores were 8.3 +/- 4.1% and 5.6 +/- 3.8%. At 1 year, the GTR treated sites were matched, in each patient, with 1 RPL site, in terms of probing attachment level (PAL 6.8 +/- 2.4 mm GTR, and 6.5 +/- 2.3 mm RPL). At this point, 24 patients took part in a supportive periodontal care program. 20 patients did not participate, and received only sporadic care by general dentists. At 5 years, all patients were reexamined. FMPS was 10.5 +/- 6.8% and FMBS 7.7 +/- 6.4%. A significant PAL loss was observed in both sites (1.2 +/- 1.4 mm GTR, 1.3 +/- 1.3 mm RPL, p < 0.0001) between 1 and 5 years. Differences in PAL loss between GTR and RPL sites were not statistically significant. Only a minority of sites (34%), however, lost PAL, while 66% remained stable. 75% of the matched sites (GTR and RPL) within the same patients were concordant in terms of PAL stability. The 23 patients in which both sites remained stable, had good oral hygiene, complied with the recall system, and did not smoke. The 10 patients in which both sites lost PAL showed deteriorating oral hygiene, did not comply with the recall system, and smoked. PAL loss in the GTR and/or RPL sites was consistently observed in patients (losers) showing PAL loss in other teeth. Losers had, in general, negative subjects characteristics, and showed a higher prevalence of tooth loss. IN CONCLUSION: (i) GTR and RPL sites showed comparable susceptibility to periodontal breakdown; (ii) stability of outcomes was consistently associated with good oral hygiene, compliance with a supportive periodontal care program, and no cigarette smoking.

Left lung allotransplantation in pig feasibility and transplanted lung function. Experimental Lung Transplant Group of Ospedale Maggiore of Milan.

Changing attitudes to animals in research and practical considerations prompted the authors to evaluate whether the pig might be a suitable substitute for dog and baboons for single left lung transplants. Twenty-nine paired pigs were used. The first transplants on 13 pairs (group 1) were done to adapt the lung transplant technique to pigs; later transplants on 16 pig pairs (group 2) were done to evaluate operative survival, and function and histological modifications of the transplanted lung in the absence of immunosuppressive treatment. Surgical and anesthetic techniques for both donor and recipient are described in detail. The survival rate in group 2 was 68%. Hemodynamic and blood gas changes were assessed during operation. PaO2 did not drop significantly after occluding the right pulmonary artery by an inflatable cuff placed around it; this suggests that the function of the transplanted lung was preserved. The pigs were put down on the third postoperative day. Vascular and bronchial anastomoses were patent and intact, but the transplanted lung was macroscopically and microscopically altered. Lung transplants can be performed in pigs and the transplanted lung seems to be capable of functioning immediately after the operation. Alteration in the lung after 3 days is probably due to rejection.