γδ T lymphocytes from cystic fibrosis patients and healthy donors are high TNF-α and IFN-γ-producers in response to Pseudomonas aeruginosa

BACKGROUND: γδ T cells have an important immunoregulatory and effector function through cytokine release. They are involved in the responses to Gram-negative bacterium and in protection of lung epithelium integrity. On the other hand, they have been implicated in airway inflammation. METHODS: The aim of the present work was to study intracytoplasmic IL-2, IL-4, IFN-γ and TNF-α production by γδ and αβ T lymphocytes from cystic fibrosis patients and healthy donors in response to Pseudomonas aeruginosa (PA). Flow cytometric detection was performed after peripheral blood mononuclear cells (PBMC) culture with a cytosolic extract from PA and restimulation with phorbol ester plus ionomycine. Proliferative responses, activation markers and receptor usage of γδ T cells were also evaluated. RESULTS: The highest production of cytokine was of TNF-α and IFN-γ, γδ being better producers than αβ. No differences were found between patients and controls. The Vγ9δ2 subset of γδ T cells was preferentially expanded. CD25 and CD45RO expression by the αβ T subset and PBMC proliferative response to PA were defective in cystic fibrosis lymphocytes. CONCLUSION: Our results support the hypothesis that γδ T lymphocytes play an important role in the immune response to PA and in the chronic inflammatory lung reaction in cystic fibrosis patients. They do not confirm the involvement of a supressed Th1 cytokine response in the pathogenesis of this disease

The matrix metalloproteinase inhibitor BB-1101 prevents experimental autoimmune uveoretinitis (EAU)

EAU is characterized by breakdown of the blood–retinal barrier and extravasation of leucocytes into retinal tissue leading to destruction of photoreceptor cells. Matrix metalloproteinases (MMP) have been implicated in trafficking of cells into tissues, but their role in inflammatory eye disease is unclear. A synthetic MMP inhibitor, BB-1101, was administered subcutaneously, from either day 0 or day 7, to Lewis rats challenged with bovine S-antigen to induce EAU. When given up to day 14, BB-1101 reduced the incidence of disease and delayed the day of onset of clinical disease. When administered from day 7 until day 21, EAU was completely abrogated. A quantitative polymerase chain reaction (PCR) assay showed an increase of both matrilysin (MMP-7), neutrophil collagenase (MMP-8) and macrophage metalloproteinase (MMP-12) in retinas from EAU animals compared with naive controls. These enzymes are produced by activated leucocytes and act on components of the basement membrane. These results therefore implicate these MMP as integral to the development of pathology in EAU

Accuracy of thoracic pedicle screw placement in scoliosis using the ideal pedicle entry point during the freehand technique

Previously, we described the ideal pedicle entry point (IPEP) for the thoracic spine at the base of the superior facet at the junction of the lateral one third and medial two thirds with the freehand technique on cadavers. Here we measured the accuracy of thoracic pedicle screw placement (Chung et al. Int Orthop 2008) on post-operative computed tomography (CT) scans in 43 scoliosis patients who underwent operation with the freehand technique taking the same entry point. Of the 854 inserted screws, 268 (31.3%) were displaced; 88 (10.3%) and 180 (21.0%) screws were displaced medially and laterally, respectively. With regard to the safe zone, 795 screws were within the safe zone representing an accuracy rate of 93%; 448 and 406 thoracic screws inserted in adolescent idiopathic and neuromuscular scoliosis showed an accuracy of 89.9 and 94%, respectively (p = 0.6475). The accuracy rate of screws inserted in the upper, middle and lower thoracic pedicles were 94.2, 91.6 and 93.7%, respectively (p = 0.2411). The results indicate that IPEP should be considered by surgeons during thoracic pedicle screw instrumentation

Observer reliability in evaluating pedicle screw placement using computed tomography

Pedicle screw insertion in spinal surgery is a demanding technique with potential risks to neurological structures, for example, within the spinal canal. Assessing screw placement in clinical practice has been performed using plain radiographs and/or mainly axial computed tomography (CT) images. Screw placement using CT image reconstructions in multiple planes has been described, but its reliability has yet to be studied. This study aimed at addressing the clinical issue of interobserver and intraobserver reliability in the use of axial and coronal CT images for the assessment of pedicle screw placement. Fifty nine pedicle screws were studied by two experienced radiologists on two separate occasions. Screw placement was classified as “in”, “out” or “questionable”. On average, 88% and 92% of the screws were classified as “in” by the first and second radiologist, respectively. Intraobserver agreement strength was almost perfect for both observers using either axial or coronal images. Interobserver agreement strength was almost perfect (axial) and substantial (coronal) in the first reading and substantial (axial, coronal) in the second reading. Assessing screw placement in more than one CT imaging plane is not only useful but reliable. Routine use may enhance reporting quality of screw placement by surgeons and radiologists