Noemi Romagnoli

Serum amyloid A, haptoglobin, and ferritin in horses with colic: Association with common clinicopathological variables and short-term outcome

Equine colic may be associated with an acute phase response (APR). Measurement of acute phase proteins (APPs) allows the detection of an APR and may help clinicians in monitoring the disease; however, the role of APPs in colic is unclear. This study aimed to evaluate the clinical usefulness of serum amyloid A (SAA), haptoglobin and ferritin in combination with an extended clinicopathological profile in equine colic. The medical records of 54 horses were retrospectively selected. Horses were grouped based on outcome (survivors vs. non-survivors), diagnosis (ischaemic/strangulating vs. non-ischaemic/non-strangulating), and treatment (medical treatment vs. surgery). Laboratory data were compared, and a logistic regression analysis was performed for outcome prediction upon admission. A high percentage of horses had abnormal SAA (29/54), haptoglobin (20/54), and ferritin (31/54) concentrations. In particular, haptoglobin was below the reference interval in 13/54 horses. Non-survivors had significantly decreased haptoglobin and increased ferritin concentrations compared with survivors. The ischaemic/strangulating group had significantly increased creatinine and ferritin and decreased haptoglobin concentrations compared with the non-ischaemic/non-strangulating group. Creatinine was the only significant predictor of mortality in the regression analysis. In conclusion, APPs including SAA, haptoglobin, and ferritin combined with clinicopathological variables may help clinicians to understand the pathogenesis of APR and underline potential complications of equine colic. The reduction in haptoglobin concentration may suggest haemolysis or muscle fibre damage; ferritin may indicate alteration in iron metabolism and tissue damage. Further prospective studies are needed to assess diagnostic and prognostic values of APPs in colic horses.

Access to cerebrospinal fluid in piglets via the cisterna magna: optimization and description of the technique

The collection of cerebrospinal fluid is necessary in order to determine its composition. It can then be used to diagnose various diseases. The aim of the study was to develop and optimize a technique for performing safe centesis for the collection of cerebrospinal fluid in piglets and its injection through the cisterna magna. The study was divided into three phases: (1) anatomical study of cadavers, (2) in vivo application of the technique and (3) observation of recovery time. The proposed technique resulted in a safe puncture of the cisterna magna. The authors identified and confirmed the correspondence of the crista occipitalis and the wings of the atlas with the external landmarks on the cadaver by means of direct radiological visualization. The punctures were performed successfully at the first attempt in 11 out of 12 anaesthetized piglets. The technique herein described provides a reproducible safe and easy route for approaching the cisterna magna for cerebrospinal fluid collection, drug administration and gene delivery.

A Comprehensive Map of CNS Transduction by Eight Recombinant Adeno-associated Virus Serotypes Upon Cerebrospinal Fluid Administration in Pigs.

Cerebrospinal fluid administration of recombinant adeno-associated viral (rAAV) vectors has been demonstrated to be effective in delivering therapeutic genes to the central nervous system (CNS) in different disease animal models. However, a quantitative and qualitative analysis of transduction patterns of the most promising rAAV serotypes for brain targeting in large animal models is missing. Here, we characterize distribution, transduction efficiency, and cellular targeting of rAAV serotypes 1, 2, 5, 7, 9, rh.10, rh.39, and rh.43 delivered into the cisterna magna of wild-type pigs. rAAV9 showed the highest transduction efficiency and the widest distribution capability among the vectors tested. Moreover, rAAV9 robustly transduced both glia and neurons, including the motor neurons of the spinal cord. Relevant cell transduction specificity of the glia was observed after rAAV1 and rAAV7 delivery. rAAV7 also displayed a specific tropism to Purkinje cells. Evaluation of biochemical and hematological markers suggested that all rAAV serotypes tested were well tolerated. This study provides a comprehensive CNS transduction map in a useful preclinical large animal model enabling the selection of potentially clinically transferable rAAV serotypes based on disease specificity. Therefore, our data are instrumental for the clinical evaluation of these rAAV vectors in human neurodegenerative diseases.

Transdermal spinal catheter placement in piglets: Description and validation of the technique.

BACKGROUND The swine species represents a perfect model for translational medicine due to its physiological and anatomical resemblance to humans. The development of techniques for spinal catheter insertion in swine is significantly useful but, at the moment, the only technique described requires laminectomy as a surgical approach. NEW METHOD The proposed techniques represent a transdermal approach for catheter placement in piglets. The study was divided into Phase I (anatomical study on 8 cadavers) and Phase II (in vivo application of the technique in 20 anaesthetised 30-day old piglets). A spinal needle was introduced between the L2 and L3 spinous processes with a ventro-cranial orientation until cerebro-spinal fluid leakage. It was then replaced with a Tuohy needle, used to introduce the catheter into the intrathecal space. Before inserting the catheter, the approximate length from the insertion point to the external projection of the Cisterna Magna was measured using the gradation markings on the device. RESULTS The technique described allowed spinal catheter placement in all piglets. In Phase I, the correct placement was confirmed using fluoroscopy while, in Phase II, cerebrospinal fluid leakage from the needle was relied on. No clinical alterations were detected either during the procedure or during the following days. COMPARISON WITH EXISTING METHOD This technique is easy and requires less skilled operators when compared to the other existing method which involves a surgical approach. Moreover, being less invasive, it potentially leads to fewer complications. CONCLUSIONS In conclusion, the technique can be performed safely in piglets, and provides an easier and less invasive approach for spinal catheter insertion.

Proteinase-activated receptor 2 expression in the intestinal tract of the horse.

Proteinase-activated receptor 2 (PAR2) is a G-protein-coupled receptor for trypsin and mast cell tryptase; it is highly expressed at the intestinal level with multiple functions, such as epithelial permeability and intestinal motility. Many proteases activate PAR2 during tissue damage, suggesting a role of the inflammatory response receptors. The aim of the study was to evaluate the distribution and expression of PAR2 in the jejunum, the ileum and the pelvic flexure, using samples collected from healthy adult horses after slaughter. Proteinase-activated receptor 2 immunoreactivity (PAR2-IR) was observed in the enterocytes, intestinal glands, the smooth muscle of the muscularis mucosae, and the longitudinal and circular muscle layers; there were no differences in the distribution of PAR2-IR in the different sections of the intestinal tract. The protein expression level showed that the relative amount of the PAR2 content in the mucosa of the intestinal tract decreased from the small to the large intestine while the PAR2 mRNA analysed showed similar values. This study provides relevant findings concerning the distribution of the PAR2 in the intestines of healthy horses and represents the starting point for evaluating the role of the PAR2 during strangulative intestinal disease and consequent systemic intestinal reperfusion/injury complications in horses in order to identify and employ antagonist PAR2 molecules.