Histopathologic Changes, Ultrastructure, and Molecular Characterization of an Adenovirus in a Sun Conure (Aratinga solstitialis).

In this case report, we describe the pathologic changes and the ultrastructural and molecular characteristics of an adenovirus in a sun conure (Aratinga solstitialis) that presented with a history of sudden death. On histologic examination, there was multifocal hepatic and splenic necrosis. Within some hepatocytes and unidentified cells in the spleen, renal interstitial fibroblasts, and ovarian stroma were intranuclear amphophilic inclusion bodies. Electron microscopy of affected tissue showed intranuclear icosahedral viral particles with an inner capsid (29.2-33.8 nm in diameter) and an outer capsid (70.2-71.7 nm in diameter). Next-generation sequencing and BLAST analysis of complementary DNA synthesized from RNA extracted from formalin-fixed tissues showed an adenovirus, designated sun conure adenovirus (SCAdv). A DNA in situ hybridization (ISH) probe, constructed from the SCAdv and similar sequences from GenBank, was also positive in the intranuclear inclusion bodies, whereas standard ISH for psittacine adenovirus 1 was negative. These results show that ancillary diagnostic testing, such as next-generation sequencing, even using formalin-fixed, paraffin-embedded tissues, along with ISH, can be useful in identifying additional, unknown viruses that show similar pathology to commonly known viruses but do not show up as positive on routine diagnostic tests.

Reporte de caso- Cambios histopatológicos, ultraestructura y caracterización molecular de un adenovirus en una cotorra solar (Aratinga solstitialis). En este reporte de caso, se describen los cambios patológicos y las características ultraestructurales y moleculares de un adenovirus en una cotorra solar (Aratinga solstitialis) que se presentó con un historial de muerte súbita. En el examen histológico, hubo necrosis hepática y esplénica multifocal. Dentro de algunos hepatocitos y células no identificadas en el bazo, los fibroblastos intersticiales renales y en el estroma ovárico se encontraron cuerpos de inclusión anfofílicos intranucleares. La microscopía electrónica del tejido afectado mostró partículas víricas intranucleares icosaédricas con una cápside interna (de 29.2 a 33.8 nm de diámetro) y una cápside externa (de 70.2 a 71.7 nm de diámetro). Mediante el análisis de secuenciación de segunda generación y por la Herramienta de Búsqueda de Alineaciones Local Básica (con siglas en inglés BLAST) del ADN complementario sintetizado a partir de ARN extraído de tejidos fijados con formalina mostraron un adenovirus, denominado adenovirus de cotorra solar (SCAdv). Se construyó una sonda de ADN para hibridación in situ (ISH), a partir de la secuencia del virus SCAdv y de secuencias similares de GenBank, que generó reacción positiva en los cuerpos de inclusión intranucleares, mientras que la hibridación in situ estándar para el adenovirus I de psitácidos fue negativa. Estos resultados muestran que las pruebas de diagnóstico complementarias, como la secuenciación de segunda generación, utilizando tejidos fijados con formalina e incluidos en parafina junto con la hibridación in situ pueden ser útiles para identificar virus adicionales desconocidos que muestran una patología similar a los virus comúnmente conocidos, pero que no se detectan con las pruebas diagnósticas de rutina.

Spontaneous haemorrhage from a left gastric artery aneurysm as a cause of acute abdominal pain.

Visceral artery aneurysms are rare, with a reported incidence of less than 2% in the general population. 1,2 Aneurysms of the left gastric artery are particularly uncommon, accounting for 4% of all visceral aneurysms. 3,4 Although the majority are discovered incidentally and can be managed conservatively, prompt treatment of those ruptured or at risk of rupture is crucial to reduce the associated morbidity. Increasing awareness of visceral artery aneurysms as a cause of spontaneous intraperitoneal haemorrhage will improve early recognition and impact on survival. We present a rare case of spontaneous rupture of a left gastric artery aneurysm.

Severe Heterotopic Ossification following Total Knee Replacement.

Although the incidence of minor heterotopic ossification is probably higher than what is usually expected, severe heterotopic ossification (HO) is an extremely rare event following total knee replacement surgery. We present the case of a 66-year-old woman who initially had achieved an excellent range of motion following bilateral uncemented rotating platform total knee replacement, before presenting with pain and loss of range of motion at 2 months after surgery. Severe HO was diagnosed on X-rays. Treatment consisted of nonoperative measures only, including physiotherapy with hydrotherapy and anti-inflammatories. She eventually regained her range of motion when seen at 8 months after operation. This case illustrates that nonoperative treatment without the use of radiotherapy or surgery can be used to safely resolve stiffness caused by HO after total knee replacement.

Pharmacokinetics and pharmacodynamics of A77 1726 and leflunomide in domestic cats.

The pharmacokinetics and pharmacodynamics of A77 1726 and leflunomide after intravenous (i.v.) and oral (p.o.) administration were evaluated in adult cats. Three treatments were administered: a single i.v. dose of A77 1726 (4 mg/kg), a single oral dose of leflunomide (4 mg/kg), and multiple oral doses of leflunomide (2 mg/kg). Mean pharmacokinetic parameter values after a single i.v. dose of A77 1726 were distribution (A) and elimination (B) intercepts (15.2 µg/mL and 34.5 µg/mL, respectively), distribution and elimination half-lives (1.5 and 71.8 h, respectively), area under the curve (AUC(0 → ∞); 3723 µg*h/mL), mean residence time (MRT; 93 h), clearance (Cl(obs); 1.1 mL/kg/h), and volume of distribution at steady state (Vd(ss); 97 mL/kg). Mean pharmacokinetic parameter values after a single oral dose of leflunomide were absorption and elimination rate constants (0.3 1/h and 0.01 1/h, respectively), absorption and elimination half-lives (2.3 and 59.1 h, respectively), AUC(0 → ∞) (3966 µg*h/mL), and maximum observed plasma concentration (C(max); 38 µg/mL). The bioavailability after a single oral dose of leflunomide was 100%. The mean ± SD A77 1726 concentration that inhibited 50% lymphocytes (EC(50) ) was 16 ± 13.5 µg/mL. The mean ± SD maximum A77 1726 concentration (EC(max)) was 61.0 ± 23.9 µg/mL.

Single photon emission computed tomography in planning unicompartmental knee arthroplasty. A prospective study examining the association between scan findings and intraoperative assessment of osteoarthritis.

The aim of this study was to determine the relationship between preoperative single photon emission computed tomography (SPECT) scan findings and intraoperative assessments of knee osteoarthritis (OA) in patients undergoing tibiofemoral unicompartmental knee arthroplasty (UKA). Fifty knees in 46 patients undergoing UKA were investigated preoperatively with a SPECT scan to confirm unicompartmental disease. There were 38 men and 12 women in the cohort with an average age of 63 years (range 44-78). The SPECT scan uptake in each compartment of the knee was graded by a single radiologist and these findings were compared with intraoperative assessments of OA (size and grade of lesion), made by two experienced surgeons, blinded to the scan findings. Significant association was demonstrated between scan findings and OA in all compartments of the knee (p<0.05), and this was strongest in the medial compartment and weakest in the patellofemoral compartment and lateral tibial plateau. We conclude that SPECT scan is a useful imaging modality in the planning of medial tibiofemoral UKA to confirm unicompartmental disease. The lower degree of association between scan findings and OA encountered in the patellofemoral compartments and lateral tibial plateau indicates that greater vigilance should be exercised in the intraoperative assessment of these areas.

Cement retrieval in minimally invasive knee surgery: the 90 degrees ball probe.

Minimally invasive knee surgery has many potential advantages as well as disadvantages. One such disadvantage in both unicompartmental and total knee arthroplasty is the inability to visualize or retrieve extruded cement. We describe the use of a new instrument, a 90 degrees ball probe, which we have used in more than 300 minimally invasive unicompartmental knee arthroplasty cases. It provides a simple, consistent, and safe method of cement retrieval.

Haemoglobin levels following unicompartmental knee arthroplasty: influence of transfusion practice and surgical approach.

A retrospective review was undertaken of preoperative and day 3 postoperative haemoglobin (Hb) levels in all unicompartmental knee arthroplasty (UKA) procedures performed by a single surgeon. Sixty-six UKAs were performed using the same prosthesis through an open approach with patella eversion. This group was compared with 212 UKAs performed using a minimally invasive approach without patella eversion, with an implant and instrumentation specifically devised for this approach. Both groups were well matched for patient demographics, surgical and anaesthetic techniques, thromboprophylaxis, and postoperative regimes. All patients received transfusions with pre-donated blood, except the unilateral minimally invasive approach group. An analysis of covariance was undertaken to examine the influence of the surgical approach and whether surgery was unilateral or bilateral, taking into account preoperative Hb levels and units of blood transfused. The average fall in Hb following UKA, adjusted for other variables, was 2.73 g/dl with an open approach compared to 1.82 g/dl with a minimally invasive approach. This difference was significant (p=0.0044). The average postoperative Hb in the minimally invasive group was 12.05 g/dl (range, 8.8 to 15.8 g/dl). Patients undergoing unilateral minimally invasive UKA are unlikely to develop symptomatic anaemia and should not be required to predonate blood or undergo transfusion.

Unicompartmental knee arthroplasty: an intermediate report of survivorship after the introduction of a new system with analysis of failures.

We describe the outcome of a series of 66 consecutive porous coated low contact stress (LCS) unicompartmental knee arthroplasty (UKA) cases performed in 52 patients for osteoarthritis (OA) by a single surgeon. Both survival, using the endpoint of revision for any cause, and knee function, using the Oxford knee score (OKS) as a validated outcome measure, were established in a retrospective review. At an average postoperative follow-up period of 5.9 years (range 5.1-6.6), there were 8 knees in patients who had died and 58 knees in those who were still living. We established the status of all knees, and prosthesis survival at 5 years was 89.7% (95% confidence interval, 81.6% to 97.7%). Technical errors were responsible for four of six failures and included progression of lateral compartment OA due to overcorrection, a medial tibial stress fracture due to poor pin placement, and a case where cement was required and poor cementing technique lead to early tibial component loosening. In the remaining 52 knees, the average preoperative OKS had improved significantly (p<0.0001) from 37.0 (range, 17-49) to a postoperative score of 20.5 (range, 13-32). We conclude that the functional results following UKA compare favourably to total knee arthropasty (TKA); however, the survivorship of this series does not match that of published reports of TKA. The introduction of a new system of UKA includes the risk of early failures due to surgeon error, even when a surgeon is competent in UKA, warranting careful surveillance during this period.

Histopathologic findings and classification of feline renal transplants.

Seventy-seven feline transplant kidney specimens, obtained from 1 to 3,183 days (9 years) after transplantation, were reevaluated histologically and classified on the basis of the Banff '97 guidelines for human renal transplant kidneys. Overall, this classification system appeared useful in detecting rejection reactions and confirmed the finding in humans that biopsies can diagnose subclinical rejection and therefore are an important diagnostic tool for the follow up of renal transplants. However, on the basis of serum creatinine values, the severity of the acute or active and chronic lesions was not accurately reflected by this scoring system. This is thought to be due to the significant differences in histologic rejection patterns, especially in acute or active rejection, in cats when compared with humans. Tubulitis, lymphocytic glomerulitis, and vasculitis, which are the main pillars of the Banff '97 acute or active rejection scoring system, are either rare or not found in cats. The presence of significant necrotizing glomerulitis and vasculitis in feline renal transplants might imply that the rejection is complicated by acute antibody-mediated rejection. Alternatively, cyclosporine toxicity also should be considered because some of these kidneys show other signs of cyclosporine toxicity. Finally, the significance of subcapsular and interlobular phlebitis, rarely described in human rejection reactions but a distinct entity in cats, is unknown. From this study, it is clear that there are significant differences in the histology of acute or active rejection between humans and cats and that a better understanding of the histologic appearance of renal allografts will be especially beneficial for treatment and prognostic purposes.

Disposition of cyclosporine after intravenous and multi-dose oral administration in cats.

The aim of this study was to evaluate the disposition of cyclosporine after intravenous (i.v.) and oral administration and to evaluate single sampling times for therapeutic monitoring of cyclosporine drug concentrations in cats. Six adult male cats (clinically intact) were used. Two treatments consisting of a single i.v. cyclosporine (1 mg/kg) and multiple oral cyclosporine (3 mg/kg b.i.d p.o. for 2 weeks) doses. Whole blood cyclosporine concentrations were measured at fixed times by high performance liquid chromatography and pharmacokinetic values were calculated. Mean values for the i.v. data included AUC (7413 ng/mL.h), t1/2 distribution and elimination (0.705 and 9.7 h, respectively), Cmax (1513 ng/mL), and Vd(ss) (1.71 L/kg). Mean values for the oral data included AUC (6243 ng/mL.h), t1/2 of absorption and elimination (0.227 and 8.19 h, respectively), and Cmax (480.0 ng/mL). Bioavailability of orally administered cyclosporine was 29 and 25% on days 7 and 14 respectively. Whole blood comment cyclosporine concentration 2 h after administration (C2) better correlated with AUC on days 7 and 14 than trough plasma concentration (C12). The rate of oral cyclosporine absorption was less than expected and there was substantial individual variation. Therapeutic drug monitoring strategies for cyclosporine in cats should be re-evaluated.

Diagnostic predictors of complications and survival after renal transplantation in cats.

OBJECTIVE: To identify preoperative diagnostic results that predict postoperative complications and survival in feline renal-transplant recipients. STUDY DESIGN: Retrospective clinical study. ANIMALS: Sixty-one feline renal allograft recipients. METHODS: Medical records for 61 consecutive cats that underwent renal allograft transplantation between January 1, 1996, and December 1, 1999, were reviewed. Age, diagnosis, body weight, body condition score, preoperative medical treatment, systolic blood pressure, packed cell volume, biochemical parameters at admission and at the time of surgery, postoperative complications, and postoperative survival were recorded. Associations of preoperative data with the occurrence of postoperative complications were determined using logistic regression. Postoperative survival was graphed using a Kaplan-Meier cumulative-survival plot. Associations of covariates with postoperative survival were analyzed using Cox proportional hazards analysis. RESULTS: Two parameters were significantly associated with occurrence of postoperative central nervous system (CNS) disorders: blood urea nitrogen concentration (odds ratio = 1.083; 95% CI = 1.018 to 1.148) and serum creatinine concentration (odds ratio = 1.8; 95% CI = 1.413 to 2.187) at the time of surgery. Postoperative survival 6 months after transplantation was 59%, though 3-year survival remained at 42%. Of all covariates investigated, only recipient age (relative hazard = 1.183; 95% CI = 1.039 to 1.334) was significantly associated with survival. CONCLUSION AND CLINICAL RELEVANCE: Standard measures of preoperative renal dysfunction do not predict postoperative survival in cats after renal transplantation, although an increase in the degree of preoperative azotemia is associated with an increased risk of CNS disorders after surgery. Increased recipient age is associated with decreased survival after renal transplantation.

Fluvastatin in combination with rad significantly reduces graft vascular disease in rat cardiac allografts.

BACKGROUND: RAD is a potent immunosuppressive agent that has been shown to be effective in preventing acute and chronic allograft rejection in animal models. The HMGCoA reductase inhibitors have been found to reduce the incidence of graft vascular disease (GVD) in heart transplant patients and in animal models. This study was designed to investigate the effects of fluvastatin or pravastatin in a rodent model of GVD produced using low doses of RAD to prevent acute rejection. METHODS: Hearts from Fisher 344 rats were heterotopically transplanted to Lewis rat recipients. RAD was administered orally at 0.5 mg/kg per day for days 0 to 14 and then 0.25 mg/kg per day for an additional 85 days to prevent acute rejection but allow for the development of GVD. Pravastatin (20 mg/kg per day) or fluvastatin (2 or 6 mg/kg per day) was added to the RAD treatment. At the end of a 100-day treatment period, the hearts were harvested for morphometric and histopathologic examinations. RESULTS: Rats treated with fluvastatin, at either dose, had a significant (P< or =0.0239) decrease in coronary arterial intimal thickening (GVD) of approximately 43%. Rats treated with pravastatin had a 22% reduction in GVD that did not reach statistical significance. Treatment with fluvastatin, but not pravastatin, decreased the degree of endomyocardial mononuclear cell infiltration seen with RAD administered alone. CONCLUSIONS: Fluvastatin significantly decreased GVD in a rat model produced using low-dose RAD immunosuppression. To a lesser extent, pravastatin also decreased GVD in this model. These data lend further support for the study of fluvastatin, pravastatin, and other HMG-CoA reductase inhibitors for the prevention of GVD in cardiac transplant patients.

Detection and confirmation of reptilian adenovirus infection by in situ hybridization.

Adenovirus infections are documented in at least 12 different species of reptiles. In contrast to their mammalian and avian counterparts reptilian adenoviruses are not well characterized as to their pathogenic potential and their ability to cause primary disease. In the diagnostic setting, fresh tissues are often not available for virus isolation, and the confirmation of reptilian adenovirus infections is dependent largely upon electron microscopy for the identification of intranuclear viral inclusions associated with histopathologic changes. The diagnosis of adenovirus infection in 2 different species of snake was confirmed by the application of DNA in situ hybridization. Using an aviadenovirus specific oligoprobe, adenoviral DNA was observed in the nuclei of hepatocytes, Kupffer cells, endothelial cells, and enterocytes. Electron microscopy of the liver confirmed the presence of intranuclear viral particles morphologically consistent with an adenovirus. DNA in situ hybridization on formalin-fixed tissues can serve as a suitable alternative to electron microscopy in the diagnosis of reptilian adenovirus infections. Both affected snakes had other concurrent diseases, suggesting that the adenovirus may not have been the primary pathogen.

Evaluation of a portocaval venograft and ameroid ring for the occlusion of intrahepatic portocaval shunts in dogs.

OBJECTIVE: To evaluate the use of a portocaval venograft and ameroid constrictor in the surgical management of intrahepatic portosystemic shunts (PSS). STUDY DESIGN: Prospective, clinical study. ANIMAL POPULATION: Ten client-owned dogs with intrahepatic PSS. METHODS: Portal pressure was measured after temporary suture occlusion of the intrahepatic PSS. In dogs with an increase in portal pressure greater than 8 mm Hg, a single extrahepatic portocaval shunt was created using a jugular vein. An ameroid ring was placed around the venograft and the intrahepatic PSS was attenuated. Transcolonic pertechnetate scintigraphy was performed before surgery, 5 days after surgery, and 8 to 10 weeks after surgery. Dogs with continued portosystemic shunting were evaluated further by laparotomy or portography. Clinical outcome and complications were recorded. RESULTS: Mean (+/- SD) portal pressure increased from 6 +/- 3 to 19 +/- 6 mm Hg with PSS occlusion; in all 10 dogs, the increase in portal pressure was greater than 8 mm Hg. There were no intraoperative complications, and, after creation of the portocaval shunt, the intrahepatic PSS could be completely ligated in 8 of 10 dogs. The final portal pressure was 9 +/- 4 mm Hg. Postoperative complications included coagulopathy and death (1 dog), ascites (3 dogs), and incisional discharge (3 dogs). Five of 8 dogs had continued portosystemic shunting at 8 to 10 weeks after surgery. Multiple extrahepatic PSS were demonstrated in 4 of these dogs. Clinical outcome was excellent in all 9 surviving dogs. CONCLUSIONS AND CLINICAL SIGNIFICANCE: The surgical technique resulted in a high incidence of multiple extrahepatic PSS. Short-term clinical results were promising, but long-term outcome must be evaluated further.

Comparison of the in vitro antiproliferative effects of five immunosuppressive drugs on lymphocytes in whole blood from cats.

OBJECTIVE: To compare the in vitro immunosuppressive effects of cyclosporine and 4 novel immunosuppressive drugs on lymphocytes in whole blood collected from healthy cats. SAMPLE POPULATION: Whole blood samples collected from 10 healthy adult domestic shorthair cats. PROCEDURE: Mitogen-stimulated lymphocyte proliferation in whole blood incubated with and without various concentrations of cyclosporine, tacrolimus, sirolimus, mycophenolic acid (MPA), or A771726 was measured by use of [3H]thymidine incorporation. Drug concentrations that resulted in a 50% inhibition of mitogen-induced proliferation (IC50) were calculated. Lymphocyte viability was determined by use of the trypan blue dye exclusion method. RESULTS: An obvious dose-response relationship for the antiproliferative effects of each drug was detected. Mean IC50 determined with concanavalin A was 46 nM for cyclosporine, 9 nM for tacrolimus, 12 nM for sirolimus, 16 nM for MPA, and 30 mM for A771726, whereas with pokeweed mitogen, mean IC50 was 33 nM for cyclosporine, 5 nM for tacrolimus, 15 nM for sirolimus, 14 nM for mycophenolic acid, and 25 mM for A771726. Mitogen-stimulated and nonstimulated lymphocytes remained viable, regardless of drug evaluated. CONCLUSIONS AND CLINICAL RELEVANCE: Tacrolimus, sirolimus, MPA, and A771726 inhibited in vitro mitogen-stimulated proliferation of feline lymphocytes in a dose-dependent manner. These novel immunosuppressive drugs may be useful for management of immune-mediated inflammatory diseases and prevention and treatment of rejection in cats that undergo organ transplantation.

Renal transplantation in cats.

Feline renal transplantation can offer long-term survival with a normal quality of life for cats with renal failure. However, it is important to remember that renal transplantation is a treatment option and not a cure. Renal transplantation is never performed on an emergency basis or prophylactically. Feline renal transplantation requires special microvascular instruments and an operating microscope. Careful patient selection and perioperative monitoring have improved the success rate. The new microemulsified form of cyclosporine is recommended for immunosuppression. As survival times have steadily improved, long-term complications, such as diabetes and neoplasia, are now being recognized.

A functional model for whole limb transplantation in the rat.

To develop a functional model for the study of whole limb transplantation, inbred Lewis rats were used as both donors and recipients. In this model, the recipient biceps femoris muscle was elevated from its distal attachment to preserve part of the adductor function of the limb after surgery. The tibial, peroneal, and sural branches of the sciatic nerve were anastomosed separately to provide faster and more precise functional recovery. For control sensory evaluation, the saphenous branches of the femoral nerve were not reattached. A flat intramedullary pin stabilized with methyl methacrylate was used to rigidly immobilize the femur. The transplanted limbs started bearing weight at 17 to 22 days. Walking on the plantar surface of the hock and adduction of the toes gradually decreased, and the rats developed a normal walking pattern. Sciatic and tibial function indexes, based on walking track analysis, correlated well with clinical observations. In this study, a new model for limb transplantation was developed that provided good and reliable sensory and ambulatory recovery.

Management of hypertension controls postoperative neurologic disorders after renal transplantation in cats.

OBJECTIVES: To determine the prevalence and describe the management of hypertension and central nervous system (CNS) complications after renal transplantation in cats. We also compared the prevalence of CNS complications between cats monitored and treated for postoperative hypertension and a previously described, historical control group of cats not monitored or treated for postoperative hypertension. STUDY DESIGN: Retrospective clinical study. ANIMALS OR SAMPLE POPULATION: A total of 34 client-owned cats that received renal allografts for the treatment of end-stage renal failure. METHODS: Medical records were reviewed. Data obtained included preoperative and postoperative systolic blood pressures, antihypertensive therapy, response to treatment, neurologic signs, and clinical outcome. The results were compared with a historical control group of feline renal allograft recipients that were neither monitored nor treated for postoperative hypertension. RESULTS: Severe postoperative hypertension occurred in 21 of 34 of cats. Hypertension was treated in all 21 cats with subcutaneously administered hydralazine which reduced systolic blood pressure to less than 170 mm Hg in 15 minutes in 20 of 21 cats; hydralazine produced hypotension in one cat and failed to control hypertension in 1 cat. After transplantation, seizures were observed in one cat and other neurologic complications (stupor, ataxia, and central blindness) were observed in three cats. The prevalence of seizures and neurologic complication-related deaths after transplantation was significantly reduced with treatment of postoperative hypertension. CONCLUSIONS AND CLINICAL RELEVANCE: Hypertension is a major contributing factor to postoperative seizure activity after renal transplantation in cats; treatment of hypertension reduces the frequency of neurologic complications.