Blinded Comparison of Palmaroproximal-palmarodistal-oblique and Dorsoproximal-palmarodistal Oblique Radiographic Projections With Contrast Medium in the Navicular Bursa in Dissected Horse Front Feet With Native Radiographs and Gross Pathology

Background: Equine palmar foot syndrome is a common cause of chronic lameness. Radiographic and ultrasound imaging are sometimes insufficient to examine the podotrochlear apparatus. MRI and CT have great diagnostic value but are expensive and are not available for all patients. Bursography might become a highly available and inexpensive diagnostic tool for this purpose.Material and Methods: To evaluate the technique, standard palmaroproximal-palmarodistal-oblique (PaPr-PaDiO ) and dorsoproximal-palmarodistal oblique (DPr-PaDiO) radiographs were acquired both before and after injecting contrast medium in the navicular bursa (NB) and were subsequently compared with gross pathology as gold standard in 48 dissected front feet. For all results a scoring system with 4 groups was introduced. Results: Compared with the gold standard, the fleiss kappa between groups were 0.349, 0.280, 0.358, and 0.455 for DPr-PaDiO, PaPr-PaDiO, DPr-PaDiO contrast enhanced (CE), and PaPr-PaDiO (CE), respectively (P < 0.001). The combined view of native images and the view of all pictures were correlated to the gold standard with a Spearman’s rank correlation coefficient of 0.373 and 0.556, respectively (P < 0.001). The Kruskal-Wallis-Test did not reveal any significant difference between standard radiographs and gross pathology group 1, 2, and 3. Contrast enhanced (CE) radiographs showed significant differences between gross pathology group 1 versus 2 and 3. Group 4 feet (gross pathology score) were remarkably well evaluated by both radiographic imaging techniques. When more than two projections were graded with findings, the respective feet were found to be moderately to severely changed in gross pathology (specificity: 100 %, sensitivity: 41.4 %). 3/13 (23.1%) group 3 and 4/19 (21.1%) group 4 specimens showed altered CE radiographs and unremarkable native radiographs. 2/13 (15.4%) group 3 and 2/19 (10.5%) group 4 feet do not have any findings in radiographs. Conclusion: In the present study, bursography with two additional CE images led to more information on the podotrochlear apparatus compared to native radiographs alone. Some cases with no changes in native radiographs will be detected in bursography, and in specimens with altered native radiographs, the diagnosis will be ensured. Therefore, bursography is a useful tool for diagnosing horses with suspected navicular disease when an MRI is not possible or the NB is injected anyway for therapeutic purposes.

Acute and subacute toxicity assessment of Madhulai Manappagu (Siddha herbal syrup formulation) in animal model

Objectives Madhulai Manappagu – a well-known sastric and widely prescribed Siddha herbal syrup formulation indicated for treating Veluppu Noi (Anaemia especially Iron deficiency Anaemia) has been in day today practice in Tamil Nadu for a quite longer decades. The syrup is a herbal preparation which has a sweet pleasant odour and a palatable taste, contain the juice of pomegranate (Punica granatum L.) as the main ingredient. Though the formulation is a fruit juice, the safety profile of the syrup is not established and is being marketed without toxicological evaluation. The study is aimed at ascertaining the acute and sub-acute toxicity assessment of Madhulai Manappagu in Wistar Albino rats. Methods The acute and sub-acute (28day repeated oral) toxicity studies were performed as per the guidelines mentioned in the Organization for Economic Cooperation and Development (OECD) 423 (adopted on December 2001) and TG 407 (adopted on October 2008) with slight modifications respectively. For acute toxicity study, three female rats were randomly selected as control; three female rats were randomly selected and were administered a single dose of 5,000 mg/kg body weight per oral route. For sub-acute (28day repeated oral) toxicity studies, three doses of test drug MM of 500 mg/kg/day (low dose), 750 mg/kg/day (intermittent dose) and 1,000 mg/kg/day (high dose) were selected for administration. Both sexes of Wistar Albino rats were randomized into four groups of 10 animals each (five males, five females). Group I was kept as control group. Group II, III and IV served as low, intermittent and high doses of MM respectively. Animals were observed for mortality, morbidity, body weight changes, feed and water intake. Haematology, clinical biochemistry, electrolytes, gross pathology, relative organ weight and histopathological examination were performed. Results In the acute toxicity study, rats showed no toxicological signs on behavior, gross pathology and body weight of rats when treated with a single dose of 5,000 mg/kg body weight per oral route. In the subacute (28 days repeated oral) toxicity study, rats have showed no significant changes on behavior, gross pathology, body weight, and hematological and biochemical parameters when treated with Madhulai Manappagu in three different doses. Conclusions The toxicity studies which include both acute and 28 days repeated (subacute) oral toxicity studies, revealed no observed adverse effect level (NOAEL) of Madhulai Manappagu in animals. Thus the safety of the drug in human usage was ensured.

A188 TARGETING RIPK2 TO TREAT INTESTINAL INFLAMMATION CAUSED BY SHIP DEFICIENCY

Background Crohn’s disease (CD) is a form of inflammatory bowel disease characterized by chronic inflammation along the gastrointestinal tract. We have described the SHIP-/- mouse model of CD-like intestinal inflammation. SHIP-/- mice develop spontaneous ileal inflammation that is characterized by villus hyperplasia and disorganization, edema, immune cell infiltration, ulceration, loss of goblet cells, and muscle wall thickening. SHIP deficiency in people with CD has been associated with a more severe and treatment-refractory disease. Interestingly, NOD2 gene variants, which are the most profound genetic associations for CD, are also associated with a more severe CD phenotype. Muramyl dipeptide (MDP) is a molecular associated microbial pattern, which activates the NOD2 signalling pathway. Recently, SHIP has been reported to play a role in the NOD2 pathway by disrupting the downstream interaction between RIPK2 and XIAP required for NOD2-mediated NFκB activation and pro-inflammatory cytokine production. Aims Based on this, we hypothesized that SHIP deficiency contributes to inflammation in CD by increasing NOD2-mediated pro-inflammatory cytokine production. Moreover, RIPK2 inhibitors will block intestinal inflammation caused by SHIP deficiency by blocking RIPK2-XIAP interactions required for NOD2 signalling and resultant pro-inflammatory cytokine production. To test this hypothesis, my aim was to determine the effect of RIPK2 inhibitors on the development of spontaneous intestinal inflammation in SHIP-/- mice. Methods 6 week-old SHIP-/- mice (and SHIP+/+ mice controls) were treated with RIPK2 inhibitors, FCG806791773 or Z1210264067, by intra-pertioneal injections every other day for 2 weeks and compared to SHIP-/- mice treated with 2% DMSO in PBS, as a vehicle control. On Day 14, distal ilea were harvested and gross pathology was assessed. Cross-sections of the ilea were H&E stained to evaluate histological damage based on villi architecture, edema, immune cell infiltration, ulceration, goblet cell loss, and muscle wall thickening. Results As expected, SHIP+/+ mice did not have gut pathology and their healthy gut phenotype was not affected by RIPK2 inhibitors. SHIP-/- mice had ileal inflammation that was evident in gross pathology and in H&E-stained tissue sections. Importantly, SHIP-/- mice treated with RIPK2 inhibitors, FCG806791773 or Z1210264067, had reduced gross pathology compared to vehicle control-treated mice. Z1210264067 treated SHIP-/- mice also had significantly lower histological damage compared to vehicle control-treated SHIP-/- mice. Conclusions RIPK2 inhibitors reduced gross and histopathology in SHIP-/- mice suggesting that RIPK2 inhibitors may be an effective treatment for people with CD, and may be particularly effective in people with CD, who harbor NOD2 risk variants and/or have low SHIP activity. Funding Agencies CCC, CIHR

Engage in Exploration: Pathology Gross Laboratory in the COVID-Era

The outbreak of Covid-19 has changed education, including the mechanism of delivery of gross pathology laboratories. Herein, we describe how we revised our preclinical gross pathology lab to a flipped model to fit with COVID-19 regulations. A series of short, session objective-driven videos are made available online. Students are expected to watch the videos before coming to the hands-on lab. Groups of 2 students enter the gross lab on a timed basis and rotate through a series of stations. At each station, students examine gross pathology specimens while answering questions designed to apply the clinical correlation of pathophysiology and heighten observational skills. One or 2 pathologists are available throughout the lab session to address the questions from the students. The design of this laboratory exercise maintains appropriate distancing and hygiene in the time of COVID-19. The laboratory rooms are mapped to set up an appropriate number of timed stations. Flow-through of the rooms is unidirectional. Comparing with the traditional show-and-tell of teaching gross pathology, the renovated flipped model is genuinely student-centered and focuses on active learning. Holding the specimen in their hands, students learn from discovery as they are completely engaged by exploring the specimen and deriving answers themselves. The flipped learning gross pathology method has been very well received and evaluated highly by both faculty and students.