Immunohistochemical Localization of RANK, RANKL and OPG in Healthy and Arthritic Canine Elbow Joints

Objective— To determine if the receptor activator of nuclear factor-κB–receptor activator of nuclear factor-κB ligand–osteoprotegerin (RANK–RANKL–OPG) system is active in bone remodeling in dogs and, if so, whether differences in expression of these mediators occur in healthy and arthritic joints.
Study Design— Experimental study.
Sample Population— Fragmented processus coronoidei (n=20) were surgically removed from dogs with elbow arthritis and 5 corresponding healthy samples from dogs euthanatized for reasons other than elbow joint disease.
Methods— Bright-field immunohistochemistry and high-resolution fluorescence microscopy were used to investigate the distribution of RANK, RANKL, and OPG in healthy and arthritic joints.
Results— All 3 molecules were identified by immunostaining of canine bone tissue. In elbow dysplasia, the number of RANK-positive osteoclasts was increased. In their vicinity, cells expressing RANKL, a mediator of osteoclast activation, were abundant whereas the number of osteoblasts having the potential to limit osteoclastogenesis and bone resorption via OPG was few.
Conclusions— The RANK–RANKL–OPG system is active in bone remodeling in dogs. In elbow dysplasia, a surplus of molecules promoting osteoclastogenesis was evident and is indicative of an imbalance between the mediators regulating bone resorption and bone formation. Both OPG and neutralizing antibodies against RANKL have the potential to counterbalance bone resorption.
Clinical Relevance— Therapeutic use of neutralizing antibodies against RANKL to inhibit osteoclast activation warrants further investigation.     

Comparison of ultrasonographic findings with clinical activity index (CIBDAI) and diagnosis in dogs with chronic enteropathies.

Intestinal wall thickness is neither a specific nor sensitive ultrasound parameter for detecting intestinal inflammation. We hypothesize that mucosal echogenicity, lymphadenomegaly, and secondary findings of the gastrointestinal tract would be more sensitive and specific markers for detecting and differentiating causes of chronic inflammatory bowel disease in dogs. Fifty-six client-owned dogs with chronic diarrhea and 10 control dogs were examined with two-dimensional, gray-scale ultrasound (time 0, 4, and 10 weeks post therapy) and small intestinal mucosal biopsies were performed at the 0- and 4-week time points. The clinical activity was assessed at each time point using the canine inflammatory bowel disease activity index (CIBDAI). Fifty-one dogs had inflammatory infiltration of the duodenal mucosa and were divided into three groups, food-responsive disease, idiopathic inflammatory bowel disease, and protein-losing enteropathy, based on their response to the different treatments and histology. Two different patterns of increased echogenicity of the mucosa were detected: hyperechoic speckles and hyperechoic striations. A normal, hypoechoic bowel mucosa in dogs with chronic diarrhea had a sensitivity of 80% and a specificity of 81% for the diagnosis of food-responsive disease. Hyperechoic striations had a sensitivity of 75% and a specificity of 96% for dogs with protein-losing enteropathy. Hyperechoic speckles were non-specific for diagnosing inflammatory bowel disease. There was a significant relationship between ultrasound score and CIBDAI at t0, but not following therapy. Mucosal echogenicity may be a better parameter for detecting inflammatory bowel disease than bowel wall thickness in dogs with chronic diarrhea.     

CONTRAST‐ENHANCED ULTRASONOGRAPHY OF THE SMALL BOWEL IN HEALTHY CATS

We characterized the pattern of ultrasonographic contrast enhancement of the small intestinal wall using a commercial contrast medium (Sonovue^®) in 10 healthy awake cats. Subjectively, a rapid intense enhancement of the serosal and submucosal layers was followed by gradual enhancement of the entire wall section during the early phase. At peak enhancement, there was a subjective loss of demarcation between intestinal wall layers. In the late phase, there was a gradual wash out of signal from the intestinal wall. Submucosal wash out occurred last. Time‐intensity curves were generated for selected regions in the intestinal wall and multiple perfusion parameters were calculated for each cat. Perfusion parameters included arrival time (7.64 ± 2.23 s), baseline intensity (1.04 ± 0.04 a.u.), time to peak from injection (10.74 ± 2.08 s), time to peak from initial rise (3.1 ± 1.15), peak intensity (8.92 ± 3.72 a.u.), wash‐in rate (2.06 ± 0.70 a.u./s) and wash‐out rate (?1.07 ± 0.91 a.u./s). The perfusion pattern of normal feline small bowel may be useful for characterizing feline gastrointestinal disorders that involve the intestinal wall.     

USE OF TRANSSPLENIC INJECTION OF AGITATED SALINE AND HEPARINIZED BLOOD FOR THE ULTRASONOGRAPHIC DIAGNOSIS OF MACROSCOPIC PORTOSYSTEMIC SHUNTS IN DOGS

We describe the use of ultrasonography‐guided percutaneous splenic injection of agitated saline and heparinized blood for the diagnosis of portosystemic shunts (PSS) in 34 dogs. Agitated saline mixed with 1 ml of heparinized autologous blood was injected into the spleen of 34 sedated dogs under sonographic guidance. The transducer was then sequentially repositioned to visualize the portal vein, the caudal vena cava, and the right atrium through different acoustic windows. It was possible to differentiate between intrahepatic and extrahepatic shunts depending on the entry point of the microbubbles into the caudal vena cava. Portoazygos shunts and portocaval shunts could be differentiated based on the presence of microbubbles in the caudal vena cava and/or the right atrium. In one dog, collateral circulation due to portal hypertension was identified. In dogs with a single extrahepatic shunt, the microbubbles helped identify the shunting vessel. The technique was also used postoperatively to assess the efficacy of shunt closure. All abnormal vessels were confirmed by exploratory laparotomy or with ultrasonographic identification of the shunting vessel. Ultrasound‐guided transsplenic injection of agitated saline with heparinized blood should be considered as a valuable technique for the diagnosis of PSS; it is easy to perform, safe, and the results are easily reproducible.     

Agglomeration Economies and Employment Growth: New Evidence from the Information Technology Sector in Italy

This paper uses local labour market area (LLMA) data to investigate the dynamics of employment in the information technology (IT) sector in Italy between 2001 and 2005. The aim is to test if agglomeration forces might significantly affect local IT employment growth. The OLS results are broadly consistent with those of earlier studies. In particular, IT employment growth is enhanced by industrial diversity (Jacobs externalities) and by plant size (economies of scale). At the same time, LLMAs with higher IT concentrations are associated with lower employment growth rates. As a robustness check, quantile regression analysis is performed. This additional set of results reveals that the role of agglomeration forces is different across IT growth levels.     

ULTRASONOGRAPHIC APPEARANCE OF THE INTRAVASCULAR TRANSIT OF AGITATED SALINE IN NORMAL DOGS FOLLOWING ULTRASOUND GUIDED PERCUTANEOUS SPLENIC INJECTION

Portosystemic shunts (PSSs) allow portal blood to bypass the liver and enter the systemic circulation. Definitive diagnosis requires surgical identification, positive contrast portography, ultrasonography, or scintigraphy. This study was designed as a preliminary step to developing an alternative/adjuvant protocol to these imaging modalities. The main goals were to establish a technique for ultrasound‐guided percutaneous trans‐splenic injection of agitated saline, to evaluate the feasibility of performing the test to explore the postsplenic portal vasculature highlighted by the microbubbles, and to ascertain whether agitated saline microbubbles cross the sinusoidal barrier. Agitated saline was injected into the spleen of 20 healthy sedated dogs under sonographic guidance. The transducer was then repositioned to visualize the portal vein, the caudal vena cava, and the right atrium through different acoustic windows. Satisfactory results were achieved in all dogs. The microbubbles were visualized in all dogs as small intense echo signals within the portal vein at the level of the porta hepatis immediately after injection. In 18 out of 20 dogs, the echogenic signal of the microbubbles disappeared immediately once within the hepatic parenchyma, whereas in two dogs, the echoes from the microbubbles lasted for several seconds within the intrahepatic portal vasculature. The absence of microbubbles beyond the sinusoidal barrier in all of the healthy dogs included in this study makes trans‐splenic injection of agitated saline a candidate as an adjuvant technique for the diagnosis of PSS, being easy to perform and repeat, as well as safe and technically feasible.     

Habitat suitability modelling for sardine juveniles (Sardina pilchardus) in the Mediterranean Sea

Identification of potential juvenile grounds of short‐lived species such as European sardine (Sardina pilchardus) in relation to the environment is a crucial issue for effective management. In the current work, habitat suitability modelling was applied to acoustic data derived from both the western and eastern part of the Mediterranean Sea. Early summer acoustic data of sardine juveniles were modelled using generalized additive models along with satellite environmental and bathymetry data. Selected models were used to construct maps that exhibit the probability of presence in the study areas, as well as throughout the entire Mediterranean basin, as a measure of habitat adequacy. Areas with high probability of supporting sardine juvenile presence persistently within the study period were identified throughout the Mediterranean Sea. Furthermore, within the study period, a positive relationship was found between suitable habitat extent and the changes in abundance of sardine juveniles in each study area.