INTERPRETATION OF INJURIES AND ASSESSMENT OF CAUSES OF DEATH IN WILD MAMMALS RECEIVED AT CETAS / IBAMA IN RIO BRANCO, ACRE, BRAZIL:

The necropsy of wild animals is necessary to raise the awareness of the competent public organizations and the population about the risks of zoonosis. Given the scarcity of information the aim of this article was to survey of the main injuries and causes of deaths of wild mammals kept in captivity was made, through the post mortem diagnosis and who passed through Wild Animal Screening Center (Centro de Triagem de Animais Silvestres - CETAS) of Rio Branco - Acre, Brazil, from September 2012 to September 2015. After death, the animals were kept refrigerated or frozen until the time of necropsy, using the standard technique for small mammals. Fragments of organs and tissues were collected, and the material was processed for histopathology using formalin fixation (10%), paraffin impregnation, hematoxylin and eosin staining, in 4 µm thick sections. 42 animals were submitted to necropsy, 27 males (64.3%) and 15 females (35.7%), of which 21 were adults (50%), 15 were puppies (35.7%) and six were young (14.3%). The main cause of death was hypovolemic shock (11.6%), followed by starvation (9.3%). There were also many deaths from undetermined causes (11.6%). A greater occurrence of deaths was registered in the Guariba monkey (Alouatta senicullus). The identification of necropsy findings and the interpretation of macroscopic lesions showed that cardiovascular lesion was the most common deaths. There does not seem to be an influence between the dry and rainy periods on the number of deaths of these animals.

Bacterial Colonization of the Condyle in Patients with Advanced Mandibular Osteoradionecrosis: Analysis of Hemimandibulectomy Specimens

Advanced mandibular osteoradionecrosis (ORN) sometimes requires extended resection (e.g., hemimandibulectomy). Bacterial infection contributes to ORN pathogenesis. To control infection and determine the extent of debridement required, an understanding of bacterial spread within sites of mandibular ORN is important. The current study used a histopathological approach to assess bacterial colonization in the mandibular condyle and elucidate possible paths of bacterial spread towards the mandibular condyle. Four hemimandibulectomy specimens were selected. Areas of bone destruction were macroscopically assessed and confirmed using hematoxylin and eosin staining. Bacterial presence within mandibular condyle was confirmed with Gram staining. Bone exposure was observed in the molar area in all specimens. Macroscopic bone destruction was apparent especially near the medial side of the cortical wall. Gram staining revealed bacterial colonization of the mandibular condyle in three of the four specimens. In conclusion, bacteria tended to spread posteriorly and through the medial side of the mandibular cortical wall. In patients with advanced ORN, the potential for bacterial colonization of the mandibular condyle should be considered during treatment.

Automated Hematoxylin and Eosin Staining with Leica AutoStainer XL v1

PURPOSE: To stain formalin fixed paraffin tissue for microscopic evaluation. Hematoxylin will stain the nuclei of the tissue blue/purple and Eosin will stain cytoplasmic elements a spectrum of brilliant pink. QUALITY CONTROL: Fixation: 10% Neutral Buffered Formalin Technique: Paraffin sections at 5 microns Control: PCRL PROCEDURE: Station #Time in StationExact Time?Solution Name185:00NoXylene175:00NoXylene165:00NoXylene150:20No100% Alcohol140:20No100% Alcohol130:20No95% Alcohol120:20No70% AlcoholWash 11:20NoWater83:30YesHematoxylinWash 20:30YesWaterWash 30:30YesWater90:10YesAcid AlcoholWash 40:30YesWater100:10YesAmmonia WaterWash 50:30YesWater112:00Yes95%70:20YesEosin60:30Yes95%50:30Yes100%40:30Yes100%30:30NoXylene21:00NoXylene11:00NoXylene RESULTS: Nuclei……………………………………………..……………Blue/Purple Cytoplasmic Elements………………………………….Shades of brilliant pink For digital storage and possible pathology review slides are scanned on the Leica Aperio slide scanner. The file output is .svs and can be viewed by downloading freely available software from Leica.

Intra-articular Injection of Bevacizumab Enhances Bone Marrow Stimulation–Mediated Cartilage Repair in a Rabbit Osteochondral Defect Model

Background: Bone marrow stimulation (BMS) via microfracture historically has been a first-line treatment for articular cartilage lesions. However, BMS has become less favorable because of resulting fibrocartilage formation. Previous studies have shown that angiogenesis blockade promotes cartilage repair. Bevacizumab is a Food and Drug Administration–approved medication used clinically to prevent angiogenesis. Hypothesis: The intra-articular injection of bevacizumab would prevent angiogenesis after BMS and lead to improved cartilage repair with more hyaline-like cartilage. Study Design: Controlled laboratory study. Methods: The dose of bevacizumab was first optimized in a rabbit osteochondral defect model with BMS. Then, 48 rabbits (n = 8/group/time point) were divided into 3 groups: osteochondral defect (defect), osteochondral defect + BMS (BMS group), and osteochondral defect + BMS + bevacizumab intra-articular injection (bevacizumab group). Rabbits were sacrificed at either 6 or 12 weeks after surgery. Three-dimensional (3D) micro–computed tomography (microCT), macroscope score, modified O’Driscoll histology scores, collagen type 2, Herovici staining, and hematoxylin and eosin staining were performed. Angiogenesis markers were also evaluated. Results: The intra-articular dose of 12.5 mg/0.5 mL bevacizumab was found to be effective without deleteriously affecting the subchondral bone. Intra-articular injection of bevacizumab resulted in significantly improved cartilage repair for the bevacizumab group compared with the BMS or the defect group based on 3D microCT, the macroscope score (both P < .05), the modified O’Driscoll histology score ( P = .0034 and P = .019 vs defect and BMS groups, respectively), collagen type 2, Herovici staining, and hematoxylin and eosin staining at 6 weeks. Cartilage in the bevacizumab group had significantly more hyaline cartilage than did that in other groups. At 12 weeks, the cartilage layer regenerated in all groups; however, the bevacizumab group showed more hyaline-like morphology, as demonstrated by microCT, histology scores ( P < .001 and .0225 vs defect and BMS groups, respectively), histology, and immunohistochemistry. The bevacizumab injection did not significantly change mRNA expressions of smooth muscle actin, vascular endothelial growth factor, or hypoxia-inducible factor-1 alpha. Conclusion: Intra-articular injection of bevacizumab significantly enhanced the quality and quantity of hyaline-like cartilage after BMS in a rabbit model. Future large-animal and human studies are necessary to evaluate the clinical effect of this therapy, which may lead to improved BMS outcomes and thus the durability of the regenerated cartilage. Clinical Relevance: The use of bevacizumab may be an important clinical adjunct to improve BMS-mediated cartilage repair.

Conversion Therapy of Intrahepatic Cholangiocarcinoma Is Associated with Improved Prognosis and Verified by a Case of Patient-Derived Organoid

This study was performed to determine the efficacy of conversion therapy in intrahepatic cholangiocarcinoma (IHCC) and explore the feasibility of cancer organoid to direct the conversion therapy of IHCC. Patient data were retrospectively reviewed in this study and cancer organoids were established using tissues obtained from two patients. A total of 42 patients with IHCC received conversion therapy, 9 of whom were downstaged successfully, and another 157 patients were initially resectable. Kaplan–Meier curves showed that the successfully downstaged patients had a significantly improved overall survival compared to those in whom downstaging was unsuccessful (p = 0.017), and had a similar overall survival to that of initially resectable patients (p = 0.965). The IHCC organoid was successfully established from one of two obtained tissues. Routine hematoxylin and eosin staining and immunohistological staining found the organoid retained the histopathological characteristics of the original tissues. Whole exome sequencing results indicated the IHCC organoid retained appropriately 87% of the variants in the original tissue. Gemcitabine and paclitaxel exhibited the strongest inhibitory effects on the cancer organoid as determined using drug screening tests, consistent with the levels of efficacy observed in the patient from whom it was derived. This study indicates that conversion therapy could improve the survival of patients with IHCC despite its low success rate, and it may be directed by cancer organoids though this is merely a proof of feasibility.