Comparison of four techniques for synoviocentesis of the equine digital flexor tendon sheath: a cadaveric study.

OBJECTIVE: To compare four different techniques for synoviocentesis of the equine digital flexor tendon sheath (DFTS). METHODS: Fifteen inexperienced operators performed each of the following injection techniques on two cadaveric limbs: Proximal (at the proximal recess of the DFTS), Axial (axial to the proximal sesamoid bone), Base (at the base of the proximal sesamoid bone), and Distal (at the pastern). The number of attempts needed before the needle was assumed to be correctly positioned into the DFTS was recorded and 10 ml of methylene blue was injected. The limbs were dissected to determine the presence of methylene blue in the DFTS, the distance between the needle entrance point and the lateral palmar or plantar (digital) nerve, the degree of subcutaneous leakage and the distance between the border of the leakage zone and the lateral digital nerve. RESULTS: The Axial (29/30) and Distal (25/30) approaches had the highest numbers of successful injections. The median number of attempts was highest for the Axial approach. The distances from the injection point and from the border of the leakage zone to the lateral digital nerve were longer for Distal and Axial approaches. CLINICAL RELEVANCE: In the hands of inexperienced operators, the Axial approach was the most successful technique for injection of the equine DFTS. Sparse subcutaneous leakage and larger distance to the nerve when using this technique might decrease the risk of in-advertent palmar or plantar digital nerve desensitisation when performing DFTS analgesia.

Activation of the cellular mitogen-activated protein kinase pathways ERK, P38 and JNK during Toxoplasma gondii invasion.

Host cell invasion is essential for the pathogenicity of the obligate intracellular protozoan parasite Toxoplasma gondii. In the present study, we evaluated the ability of T. gondii tachyzoites to trigger phosphorylation of the different mitogen-activated protein kinases (MAPK) in human monocytic cells THP1. Kinetic experiments show that the peak of extracellular-signal-regulated kinase (ERK1/2), P38 and cjun-NH2 terminal kinase (JNKs) phosphorylation occurs between 10 and 60 min. The use of specific inhibitors of ERK1/2, P38 and JNK1/2 phosphorylation indicates the specificity of MAPKs phosphorylation during invasion. Signaling through cellular and parasite mitogen-activated protein (MAP) kinase pathways appears to be critical for T. gondii invasion.

Calmodulin distribution and the actomyosin cytoskeleton in Toxoplasma gondii.

The gliding motility of the protozoan parasite Toxoplasma gondii and its invasion of cells are powered by an actin-myosin motor. We have studied the spatial distribution and relationship between these two cytoskeleton proteins and calmodulin (CaM), the Ca(2+)-dependent protein involved in invasion by T. gondii. A 3D reconstruction using labeling and tomographic studies showed that actin was present as a V-like structure in the conoidal part of the parasite. The myosin distribution overlapped that of actin, and CaM was concentrated at the center of the apical pole. We demonstrated that the actomyosin network, CaM, and myosin light-chain kinases are confined to the apical pole of the T. gondii tachyzoite. MLCK could act as an intermediate molecule between CaM and the cytoskeleton proteins. We have developed a model of the organization of the actomyosin-CaM complex and the steps of a signaling pathway for parasite motility.

Interferon-gamma signal transduction during parasite infection: modulation of MAP kinases in the infection of human monocyte cells (THP1) by Toxoplasma gondii.

We assayed mitogen-activated protein (MAP) kinase phosphorylation in a human monocyte cell line (THP1) during their infection by Toxoplasma gondii. In addition, we tested the effect of specific MAP kinase inhibitors (PD098059 and SB203580) on parasite invasion. MAP kinase phosphorylation was increased in the cytosol and membrane fractions of THP1 infected with T. gondii. The MAP kinase phosphorylation of uninfected THP1 cells was not significantly modified by incubation for 20 h with 1000 U/ml of IFN-gamma. However, IFN-gamma treatment of infected cells significantly reduces the increase in phosphorylation caused by parasite infection. There was also MAP kinase activity in the cytosol and membrane fractions of extracellular T. gondii tachyzoites. IFN-gamma altered the distribution of activity in subcellular fractions of extracellular T. gondii tachyzoites. This indicates that IFN-gamma directly affects parasite MAP kinase activity. The results provide evidence that MAP kinase pathways participate in the infection by T. gondii and that the decrease in MAP kinase activity in infected cells caused by IFN-gamma may be involved in mediating their protective signals.