Intra-tumoral gene delivery of feIL-2, feIFN-gamma and feGM-CSF using magnetofection as a neoadjuvant treatment option for feline fibrosarcomas: a phase-I study

Despite aggressive pre- or postoperative treatment, feline fibrosarcomas have a high relapse rate. In this study, a new treatment option based on immune stimulation by intra-tumoral delivery of three feline cytokine genes was performed. The objective of this phase-I dose-escalation study was to determine a safe dose for further evaluation in a subsequent phase-II trial. Twenty-five client-owned cats with clinical diagnosis of fibrosarcoma - primary tumours as well as recurrences - entered the study. Four increasing doses of plasmids coding for feIL-2, feIFN-gamma or feGM-CSF, respectively, were previously defined. In groups I, II, III and IV these doses were 15, 50, 150 and 450 microg per plasmid and a corresponding amount of magnetic nanoparticles. Two preoperative intra-tumoral injections of the magnetic DNA solution were followed by magnetofection. A group of four control cats received only surgical treatment. Side effects were registered and graded according to the VCOG-CTCAE scale and correlated to treatment. Statistical analyses included one-way anova, post hoc and Kruskal-Wallis tests. ELISA tests detecting plasma feIFN-gamma and plasma feGM-CSF were performed. One cat out of group IV (450 microg per plasmid) showed adverse events probably related to gene delivery. As these side effects were self-limiting and occurred only in one of eight cats in group IV, this dose was determined to be well tolerable. Altogether six cats developed local recurrences during a 1-year observation period. Four of these cats had been treated with dose IV. Regarding these observations, a subsequent phase-II trial including a representative amount of cats should be tested for the efficacy of dose IV as well as dose III.     

Morphometric analysis of the corneal endothelium of Yacare caiman (Caiman yacare) using scanning electron microscopy

The objective of this study was to examine the endothelial surface morphology and to perform morphometric analysis of the corneal endothelial cells of Yacare caiman (Caiman yacare) using scanning electron microscopy. Morphometric analysis with regard to polygonality, mean cell area, cell density and coefficient of variation of mean cell area was performed. Cell areas were measured using image analysis software. The normal corneal endothelium of Yacare caiman consisted of polygonal cells of uniform size and shape with interdigitations of the cell borders. Microvilli appeared as protrusions on the cellular surface. The average cell area was 270 +/- 24 microm(2) and the endothelial cell density was 3704 +/- 324 cells/mm(2). The coefficient of variation of cell area was 0.22. This study demonstrates that the Yacare caiman corneal endothelium is similar to those described in other vertebrates.     

Evaluation of a focused assessment with sonography for trauma protocol to detect free abdominal fluid in dogs involved in motor vehicle accidents

OBJECTIVE: To establish a focused assessment with sonography for trauma (FAST) protocol in dogs, determine whether FAST can be performed by veterinary clinicians without extensive ultrasonographic experience, and assess the frequency of free fluid (as determined via FAST) in the abdominal cavity of dogs following motor vehicle accidents (MVAs). DESIGN: Prospective study. ANIMALS: 100 client-owned dogs evaluated within 24 hours of an MVA. PROCEDURE: Dogs were placed in lateral recumbency for the FAST examination. To detect fluid in the abdomen, 2 ultrasonographic views (transverse and longitudinal) were obtained at each of 4 sites (just caudal to the xiphoid process, on the midline over the urinary bladder, and at the left and right flank regions). RESULTS: In the 100 dogs evaluated via FAST, free abdominal fluid was detected in 45 dogs. In 40 of those 45 dogs, abdominocentesis was performed; hemoperitoneum and uroperitoneum were diagnosed in 38 and 2 dogs, respectively. Compared with dogs that had no free abdominal fluid detected via FAST, dogs that had free abdominal fluid detected via FAST had significantly higher heart rates and serum lactate concentrations and significantly lower PCVs and total solid concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that FAST is a simple and rapid technique that can be performed on dogs in an emergency setting to detect intra-abdominal free fluid and can be performed by veterinary clinicians with minimal previous ultrasonographic experience.     

CliniTox: the computer-based information system for poisoning in farm animals

Poisonings can cause reduced performance and severe economic loss in farm animals. A swift and targeted action is required from veterinarians. We have established a computer-based decision support system for poisonings in ruminants and pigs. The system offers access to the most recent information available in clinical toxicology of farm animals. Towards this goal, relevant reports from the peer-reviewed literature were evaluated and organised according to the requirements of a structured database. The information provided for each toxic substance includes a summary of its chemical and physical properties, sources, commercial uses or natural occurrences, toxicokinetic data, mechanisms of action, threshold doses, clinical symptoms with brief case reports, sampling and analytical results, post-mortem abnormalities, differential diagnoses, therapeutic guidelines and references to the literature. This decision support system has been programmed with two user-friendly search functions: a search tool that allows the choice of clinical and pathological findings, and another function that serves to find a substance using its chemical name, the class of compounds to wich it belongs, a possible source or one of its main applications. CliniTox can be accessed directly via our webserver (http://www.clinitox.ch).     

An investigation into reduced milk production following dietary alteration on an Irish dairy farm

A nutritional evaluation of an Irish dairy herd indicated gross overfeeding of late lactation cows, over-conditioning of cows at parturition and a high rate of body condition loss in early lactation. Metabolisable-energy based nutritional modelling software was used to guide recommended dietary changes to prevent excessive condition gain in late lactation. Immediately following the implementation of the changes there was an unexpected reduction in performance affecting both milk yield and protein concentration. An investigation into the poor performance revealed underestimation of peak milk yield; over-estimation of maize silage quality; a large difference in the concentrate being fed compared to the concentrate recommended, and failure of the blend of concentrate ingredients to maintain the intended proportions in the in-parlour feeding system. The estimated maximum cumulative effect of these errors was to cause undersupply of energy and protein in the recommended diet of 16% and 3% respectively to cows in early lactation. Use of a net-energy nutritional model would have indicated a requirement for a higher energy supply in this case. This report highlights the challenges in obtaining accurate on-farm data for use in dairy cow nutritional models.     

Effect of Estradiol-17β on protein synthesis and degradation rates in fused bovine satellite cell cultures

Although androgenic and estrogenic steroids are widely used to enhance muscle growth and increase feed efficiency in feedlot cattle, their mechanism of action is not well understood. Further, in vivo studies indicate that estradiol (E2) affects muscle protein synthesis and/or degradation, but in vitro results are inconsistent. We have examined the effects of E2 treatment on protein synthesis and degradation rates in fused bovine satellite cell (BSC) cultures. Additionally, to learn more about the mechanisms involved in E2-enhanced muscle growth, we have examined the effects of compounds that interfere with binding of E2 or insulin-like growth factor (IGF)-1 to their respective receptors on E2-induced alterations in protein synthesis and degradation rates in BSC cultures. Treatment of fused BSC cultures with E2 results in a concentration-dependent increase (P < 0.05) in protein synthesis rate and a decrease (P < 0.05) in protein degradation rate. The pure estrogen antagonist ICI 182 780 suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation in fused BSC cultures. The G-protein coupled receptor (GPR)-30 agonist G1 does not affect either synthesis or degradation rate, which establishes that GPR30 does not play a role in E2-induced alterations in protein synthesis or degradation. JB1, a competitive inhibitor of IGF-1 binding to the Type 1 insulin-like growth factor receptor (IGFR-1), suppresses (P < 0.05) E2-induced alterations in protein synthesis and degradation. In summary, our data show that E2 treatment directly alters both protein synthesis and degradation rates in fused BSC cultures via mechanisms involving both the classical estrogen receptor (ER) and IGFR-1.