Relationship between clinical signs and pulmonary function estimated by the single breath diagram for CO(2) (SBD-CO(2)) in horses with chronic obstructive pulmonary disease

The pulmonary health of 66 horses was assessed by a clinical examination and simple supplementary diagnostic methods. Single breath diagrams for CO(2) (SBD-CO(2)) and derived lung function indices were used to determine pulmonary function. The clinical signs in different groups were related to the results of the lung function indices derived from the SBD-CO(2). In horses with moderate to severe chronic obstructive pulmonary disease (COPD), a significant relationship was found between the respiratory frequency and the ratio of Bohr's dead space to the tidal volume (VD(Bohr)/VT), and between the physiological dead space/tidal volume ratio (VD(phys)/VT) and the ratio of the alveolar dead space to the alveolar tidal volume (VD(alv)/VT(alv)), but no significant associations were found between the arterial oxygen tension (P(a)O(2)) and lung function indices derived from the SBD-CO(2). The occurrence of cough, the viscosity of tracheobronchial mucus and the amount of polynuclear neutrophils in tracheobronchial aspirates were significantly related to the expiratory tidal volume (VT), the total expired volume of CO(2) (VCO(2)), VD(Bohr)/VT, VD(phys)/VT and VD(alv)/VT(alv).We conclude that abnormal findings in these clinical parameters indicate a measurable ventilation and perfusion (V(A)/Q) mismatch which is reflected by increases in dead space, VD(Bohr)/VT and VD(phys)/VT as well as VD(alv)/VT(alv).     

Method for co-purification of equine neutrophil elastase and myeloperoxidase from a limited blood volume

Neutrophil myeloperoxidase (MPO) and elastase can be released in severe inflammatory diseases and cause tissue injuries. Equine enzymes have already been individually purified from large blood quantities. We describe the isolation of both enzymes from a same limited blood volume. Both MPO and elastase were extracted by crushing PMN isolated by centrifugation on a percoll-gradient from a 460 ml blood collection. MPO and elastase were separated by an ionic exchange chromatography phase and further purified by gel filtration chromatography on Superdex 200 and 75, respectively. Enzymes were identified in the collected fractions by specific enzymatic assays. The final purity was verified by electrophoresis. Specific activity was improved to 19.92 and 34.3× for elastase (final yield: 340 μg) and MPO (final yield: 130 μg), respectively, during the procedure. Results show the possibility of isolating both enzymes from the same blood sample with a sufficient yield and purity for future studies on their implication and interaction during inflammatory diseases.     

Recombinant proteins and peptides as tools for studying IgE reactivity with low-molecular-weight glutenin subunits in some wheat allergies

Two genes of wheat low-molecular-weight glutenin subunits (LMW-GS), B16 and P73, were cloned and expressed in E. coli. They were homologous to proteins encoded respectively at Glu-B3 and Glu-D3 loci. The N-terminal and C-terminal halves of B16 (NB16 and B16C) and the two chimeras combining the halves of the two genes (B16-P73 and P73- B16) were also expressed. All these constructs were compared for their reactivity with IgE from 24 patients suffering from different forms of wheat allergies. The results confirmed that LMW-GSs bound IgE in all adult allergies tested. Strong differences in reactivity between all the constructs were observed. They were disease-dependent. In wheat-dependent exercise-induced anaphylaxis (WDEIA), the reactivity of the constructs depended partly on common epitopes with omega-5 gliadins but also on differences in molecule conformation. The presence of NB16 in the constructs greatly influenced their IgE reactivity.     

GnRHa‐induced Multiple Spawns and Volition Spawning of Captive Spotted Rose Snapper,Lutjanus guttatus,at Mazatlan,Mexico

Sexual maturation and induced spawning treatments were carried out with captive spotted rose snapper, Lutjanus guttatus. A total of 3013 × 10⁶ eggs (64.7% were floating) were produced from eight treated females in 42 spawns induced with GnRHa implants during the course of the present study. GnRHa ethylene‐vinyl acetate copolymer effective doses were 204 ± 11 µg/kg in June 2005, and 224 ± 13 µg/kg in July 2005. General fertilization was 50.9 ± 34.5% and 12–14 h after spawning, viability of floating eggs was 90.4 ± 12.4%. Mean incubation period at 29–31 C was 18–20 h, and mean hatching was 94.4 ± 8.2% (73–100%). Newly hatched larvae were 2.18 ± 0.15 mm in total length (TL). One month after the last hormone experiment, previously GnRHa‐treated and untreated fish began spawning voluntarily. Hormone‐treated breeders had higher fecundity than untreated fish, producing 72.5 million eggs versus 13.9 million eggs for the untreated fish, over the following 11 mo. Combined data of volitional spawning for total egg fertilization, viability, hatching, and larval TL were 77.7 ± 1.8%, 90.3 ± 1.3%, 87.9 ± 2%, and 2.50 ± 0.12 mm, respectively. These results can ensure the sustainability of a commercial hatchery.