Limb‐sparing in dogs using patient‐specific,three‐dimensional‐printed endoprosthesis for distal radial osteosarcoma: A pilot study

Limb‐sparing for distal radial osteosarcoma has a high rate of complications. Using personalized three‐dimensional (3D)‐printed implants might improve outcome. The goals of this study were to optimize use of patient‐specific, 3D‐printed endoprostheses for limb‐sparing in dogs in the clinical environment and to report the outcome. This was a pilot study where five client‐owned dogs were enrolled. Computed tomography (CT) of the thoracic limbs was performed, which was used to create patient‐specific endoprostheses and cutting guides, and repeated on the day of surgery. Intra‐arterial (IA) carboplatin was introduced in the clinical management. Limb‐sparing was performed. Outcome measures were time required to produce the endoprosthesis and cutting guide, fit between cutting guide and endoprosthesis with host bones, gait analysis, size of the tumour, percent tumour necrosis, complications, disease‐free interval (DFI) and survival time (ST). Four dogs received IA carboplatin. Excessive tumour growth between planning CT and surgery did not occur in any dog. The interval between the CT and surgery ranged from 14 to 70 days. Fit between the cutting‐guide and endoprosthesis with the host bones was good to excellent. At least one complication occurred in all dogs. Two dogs were euthanized with STs of 192 and 531 days. The other dogs were alive with a follow up of 534 to 575 days. IA chemotherapy is a promising strategy to minimize the risk of excessive tumour growth while waiting for the endoprosthesis and cutting‐guide to be made. The design of the cutting‐guide was critical for best fit of the endoprosthesis with host bones.     

Pilot study to evaluate the efficacy of lymphotropic nanoparticle enhanced MRI for diagnosis of metastatic disease in canine head and neck tumours

This pilot study is designed to determine if lymphotropic nanoparticle enhanced MRI (LNMRI) is a viable technique for staging of naturally occurring canine malignant head and neck tumours. Previous imaging studies in veterinary medicine have shown variable sensitivity and specificity for determining metastasis for local lymph nodes in head and neck tumours. LNMRI utilizes ultra‐small superparamagnetic iron oxide nanoparticles (USPIOs) to help in the detection of metastatic disease in lymph nodes. USPIOs are phagocytized and localized to normal lymph nodes where they assist in evaluation for regions of effacement by cancerous cells. Six dogs underwent LNMRI for the diagnosis of metastatic lymph nodes. A truncated MRI consisting of transverse images of T2, T1 pre‐ and post‐contrast and T2* sequences were evaluated for presence of metastasis. Sentinel lymph nodes and lymph nodes with possible metastatic lesions were surgically excised for histological evaluation. In the initial phase of this study, 24 lymph nodes were included in analysis. Subjective observation by the primary investigator had a calculated sensitivity and specificity of 100% and 88% based on histological results. There were no negative side effects to the USPIOs noted in the limited number of patients in this study. Percentage signal intensity loss was calculated and found to be significantly different between metastatic and non‐metastatic lymph nodes (P‐value = .038). In conclusion, this pilot study shows that LNMRI has the potential to be a sensitive and specific method of diagnosing lymph node metastasis. Further research is warranted to determine if this method is clinically applicable and accurate.     

Localized stem chilling alters carbon processes in the adjacent stem and in source leaves

Transport phloem is no longer associated with impermeable pipes, but is instead considered as a leaky system in which loss and retrieval mechanisms occur. Local stem chilling is often used to study these phenomena. In this study, 5-cm- lengths of stems of 3-year-old oak trees (Quercus robur L.) were locally chilled for 1 week to investigate whether observations at stem and leaf level can be explained by the leakage-retrieval mechanism. The chilling experiment was repeated three times across the growing season. Measurements were made of leaf photosynthesis, carbohydrate concentrations in leaves and bark, stem growth and maximum daily stem shrinkage. Across the growing season, a feedback inhibition in leaf photosynthesis was observed, causing increased dark respiration and starch concentration. This inhibition was attributed to the total phloem resistance which locally increased due to the cold temperatures. It is hypothesized that this higher phloem resistance increased the phloem pressure above the cold block up to the source leaves, inducing feedback inhibition. In addition, an increase in radial stem growth and carbohydrate concentration was observed above the cold block, while the opposite occurred below the block. These observations indicate that net lateral leakage of carbohydrates from the phloem was enhanced above the cold block and that translocation towards regions below the block decreased. This behaviour is probably also attributable to the higher phloem resistance. The chilling effects on radial stem growth and carbohydrate concentration were significant in the middle of the growing season, while they were not at the beginning and near the end of the growing season. Furthermore, maximum daily shrinkages were larger above the cold block during all chilling experiments, indicating an increased resistance in the xylem vessels, also generated by low temperatures. In conclusion, localized stem chilling altered multiple carbon processes in the source leaves and the main stem by changing hydraulic resistances.     

Rapid analysis of seed size in <Emphasis Type="Italic">Arabidopsis</Emphasis> for mutant and QTL discovery

Background  

Arabidopsis thaliana is a useful model organism for deciphering the genetic determinants of seed size; however the small size of its seeds makes measurements difficult. Bulk seed weights are often used as an indicator of average seed size, but details of individual seed is obscured. Analysis of seed images is possible but issues arise from variations in seed pigmentation and shadowing making analysis laborious. We therefore investigated the use of a consumer level scanner to facilitate seed size measurements in conjunction with open source image-processing software.     

Identification of an N-terminal formylated, two-peptide bacteriocin from Enterococcus faecalis 710C

Enterococcus faecalis 710C, isolated from beef product, has a broad antimicrobial activity spectrum against foodborne pathogens. Two bacteriocins, enterocin 7A (Ent7A) and enterocin 7B (Ent7B), were purified from the culture supernatant of E. faecalis 710C and characterized using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry and electrospray infusion tandem mass spectrometry analyses. These data and subsequent genetic analysis showed that Ent7A and Ent7B are produced without N-terminal leader sequences and have amino acid sequences that are identical to those of enterocins MR10A and MR10B, respectively. However, the observed masses for Ent7A and Ent7B are 5200.80 and 5206.65 Da (monoisotopic mass), respectively, which are higher than the theoretical molecular masses of MR10A and MR10B, respectively. This study provides evidence that both Ent7A and Ent7B are formylated on the N-terminal methionine residue. Purified Ent7A and Ent7B are active against spoilage microorganisms and foodborne pathogens, including Clostridium sporogenes , Listeria monocytogenes , and Staphylococcus aureus as well as Brevundimonas diminuta , which has been associated with infections among immune-suppressed cancer patients.     

Web-based documentation of clinical skills to assess the competency of veterinary students

Kansas State University implemented a Web-based program to assess students' competency to perform technical skills during clinical rotations throughout the fourth year of the veterinary curriculum. The classes of 2009 and 2010 recorded a minimum number of procedures (104 and 103, respectively) from a menu of more than 220 recommended procedures. Procedures were categorized by species (small animal, equine, food animal) and disciplines (imaging, anesthesia, diagnostic medicine/necropsy). Ophthalmology was added as a fourth discipline for the class of 2010. Students recorded procedures into the Web-based system, including information about the patient, procedure performed, supervisor, and a self-assessment of performance. Faculty, staff, and house officers evaluated the procedures electronically by confirming that they witnessed the procedure and providing qualitative and written feedback. The class of 2009 recorded 18,492 procedures (M=171/student) and the class of 2010 recorded 16,935 procedures (M=158/student). Two students from each class (2009 and 2010) did not complete the minimum required skills during clinical rotations and returned to perform procedures immediately before (n=3) or immediately after (n=1) graduation to receive their diploma. The Web-based system captured a large number of assessments of technical competency performed in the clinical setting. The system provided students with formative feedback throughout the clinical year, ensured equitable distribution of procedural opportunities across the student body, and required minimal additional resources.     

Comparative evaluation of molecular diagnostic tests for Nucleospora salmonis and prevalence in migrating juvenile salmonids from the Snake River, USA

Nucleospora salmonis is an intranuclear microsporidian that primarily infects lymphoblast cells and contributes to chronic lymphoblastosis and a leukemia-like condition in a range of salmonid species. The primary goal of this study was to evaluate the prevalence of N. salmonis in out-migrating juvenile hatchery and wild Chinook salmon Oncorhynchus tshawytscha and steelhead O. mykiss from the Snake River in the U.S. Pacific Northwest. To achieve this goal, we first addressed the following concerns about current molecular diagnostic tests for N. salmonis: (1) nonspecific amplification patterns by the published nested polymerase chain reaction (nPCR) test, (2) incomplete validation of the published quantitative PCR (qPCR) test, and (3) whether N. salmonis can be detected reliably from nonlethal samples. Here, we present an optimized nPCR protocol that eliminates nonspecific amplification. During validation of the published qPCR test, our laboratory developed a second qPCR test that targeted a different gene sequence and used different probe chemistry for comparison purposes. We simultaneously evaluated the two different qPCR tests for N. salmonis and foundthat both assays were highly specific, sensitive, and repeatable. The nPCR and qPCR tests had good overall concordance when DNA samples derived from both apparently healthy and clinically diseased hatchery rainbow trout were tested. Finally, we demonstrated that gill snips were a suitable tissue for nonlethal detection of N. salmonis DNA in juvenile salmonids. Monitoring of juvenile salmonid fish in the Snake River over a 3-year period revealed low prevalence of N. salmonis in hatchery and wild Chinook salmon and wild steelhead but significantly higher prevalence in hatchery-derived steelhead. Routine monitoring of N. salmonis is not performed for all hatchery steelhead populations. At present, the possible contribution of this pathogen to delayed mortality of steelhead has not been determined.