Abdominal wall mass and hemoabdomen in a Haflinger mare.

A 6-year-old Haflinger mare was presented with a history of recurrent hemoabdomen. On necropsy, a firm infiltrative multinodular yellow mass was observed in the wall of the posterior abdomen. Histopathologic examination revealed a proliferation of fibroblastic cells, which were positive for α-smooth muscle actin and vimentin.

Detection of rib trauma in newborn foals in an equine critical care unit: a comparison of ultrasonography, radiography and physical examination.

REASONS FOR PERFORMING STUDY: Previous studies have shown that in man ultrasonography is more accurate than radiography for detecting rib fractures. OBJECTIVES: To describe clinical, radiographic and ultrasonographic findings related with rib fractures in newborn foals in an equine critical care unit; and to compare diagnostic accuracy of ultrasonography to radiography. METHODS: A prospective ultrasonographic study was performed on 29 foals presented to the emergency unit. This study was performed at the Centre Hospitalier Universitaire Vétérinaire (CHUV), University of Montreal. Physical examination as well as radiographic and ultrasonographic examinations were performed. RESULTS: Thoracic radiographs revealed 10 rib fractures in 5 of 26 (19%) foals. Ultrasonography revealed 49 fractures in 19 of 29 (65%) foals of which fillies (n = 13; 68%) were significantly over represented as were fractures to the left thorax (n = 15; 78%). Seventeen of 19 foals (90%) had rib fractures located 3 cm or less from the costochondral junction, the distal part of the rib being displaced laterally in all cases. In 2 foals, where both thoracic radiographs and ultrasonography detected rib fractures, the site of fractures was located on the mid portion of the rib. Rib fractures were detected only by thoracic radiographs in one foal. Sixty-five percent (32/49) of fractured ribs had a moderate displacement (1-4 mm). CONCLUSIONS: Rib fractures are seen frequently in newborn foals in equine critical care units. Ultrasonography is more accurate than radiography and reveals fractures in most patients presented in emergency. The position (costochondral junction) of rib fractures and of the fragments suggest that most thoracic trauma probably occurs during parturition. POTENTIAL RELEVANCE: Ultrasound imaging increases awareness and improves the diagnosis of rib fractures in newborn foals.

Implementation of semi-automated cloning and prokaryotic expression screening: the impact of SPINE.

The implementation of high-throughput (HTP) cloning and expression screening in Escherichia coli by 14 laboratories in the Structural Proteomics In Europe (SPINE) consortium is described. Cloning efficiencies of greater than 80% have been achieved for the three non-ligation-based cloning techniques used, namely Gateway, ligation-indendent cloning of PCR products (LIC-PCR) and In-Fusion, with LIC-PCR emerging as the most cost-effective. On average, two constructs have been made for each of the approximately 1700 protein targets selected by SPINE for protein production. Overall, HTP expression screening in E. coli has yielded 32% soluble constructs, with at least one for 70% of the targets. In addition to the implementation of HTP cloning and expression screening, the development of two novel technologies is described, namely library-based screening for soluble constructs and parallel small-scale high-density fermentation.

Eukaryotic expression: developments for structural proteomics.

The production of sufficient quantities of protein is an essential prelude to a structure determination, but for many viral and human proteins this cannot be achieved using prokaryotic expression systems. Groups in the Structural Proteomics In Europe (SPINE) consortium have developed and implemented high-throughput (HTP) methodologies for cloning, expression screening and protein production in eukaryotic systems. Studies focused on three systems: yeast (Pichia pastoris and Saccharomyces cerevisiae), baculovirus-infected insect cells and transient expression in mammalian cells. Suitable vectors for HTP cloning are described and results from their use in expression screening and protein-production pipelines are reported. Strategies for co-expression, selenomethionine labelling (in all three eukaryotic systems) and control of glycosylation (for secreted proteins in mammalian cells) are assessed.

Gene sequence and crystal structure of the aldehyde oxidoreductase from Desulfovibrio desulfuricans ATCC 27774.

The aldehyde oxidoreductase (MOD) isolated from the sulfate reducer Desulfovibrio desulfuricans (ATCC 27774) is a member of the xanthine oxidase family of molybdenum-containing enzymes. It has substrate specificity similar to that of the homologous enzyme from Desulfovibrio gigas (MOP) and the primary sequences from both enzymes show 68 % identity. The enzyme was crystallized in space group P6(1)22, with unit cell dimensions of a=b=156.4 A and c=177.1 A, and diffraction data were obtained to beyond 2.8 A. The crystal structure was solved by Patterson search techniques using the coordinates of the D. gigas enzyme. The overall fold of the D. desulfuricans enzyme is very similar to MOP and the few differences are mapped to exposed regions of the molecule. This is reflected in the electrostatic potential surfaces of both homologous enzymes, one exception being the surface potential in a region identifiable as the putative docking site of the physiological electron acceptor. Other essential features of the MOP structure, such as residues of the active-site cavity, are basically conserved in MOD. Two mutations are located in the pocket bearing a chain of catalytically relevant water molecules. As deduced from this work, both these enzymes are very closely related in terms of their sequences as well as 3D structures. The comparison allowed confirmation and establishment of features that are essential for their function; namely, conserved residues in the active-site, catalytically relevant water molecules and recognition of the physiological electron acceptor docking site.