Diff-Quik® staining method for detection and identification of monosodium urate and calcium pyrophosphate crystals in synovial fluids

OBJECTIVE To evaluate whether the Diff Quik (DQ) staining method might prove useful in identifying monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals on permanent mounted stained slides. METHODS 27 synovial fluid (SF) samples obtained from the knees of 21 patients with acute CPPD disease and 6 with acute gout were studied. Wet analysis for crystal detection and identification was performed within one hour of joint aspiration. In addition, 16 inflammatory synovial effusions obtained from patients with knee arthritis induced by non-crystalline inflammatory diseases were studied. For each SF, a DQ stained slide was analysed by two of the authors trained in SF analysis. The observers were blinded to the type of crystals present in the SF. Each slide was analysed by compensated polarised as well as transmitted light microscopy. An SF was considered positive if intracellular and/or extracellular crystals were clearly identified. In addition, the observer was asked to identify the type of the crystals using compensated polarised light microscopy. Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) of the DQ staining method were determined. RESULTS 51 true positive and 28 true negative cases were correctly classified (39 CPPD samples, 12 MSU samples, 28 samples of crystal unrelated arthropathies). Overall, four false positive and three false negative cases were reported. In all the false positive cases, extracellular CPPD crystals were erroneously identified, whereas CPPD crystals present in the SF were not identified in the three false negative cases. All MSU specimens were correctly diagnosed. The overall specificity, sensitivity, and accuracy using DQ stained slides for crystal confirmation were respectively 87.5%, 94.4%, and 91.9%. The PPV was 92.7% and the NPV 90.3%. In particular, the specificity, sensitivity, and accuracy for CPPD detection were 90.9%, 92.9%, and 91.9%, with a PPV of 90.7 and an NPV of 93.0%. All the MSU specimens were correctly identified, providing 100% sensitivity, specificity, accuracy, PPV, and NPV. CONCLUSIONS Stained preparations of SF, including DQ stained smears, could provide a useful tool for delayed SF analysis suitable for quality controls, including cytological examination and crystals detection and identification

Diff-Quik® staining method for detection and identification of monosodium urate and calcium pyrophosphate crystals in synovial fluids

The aim of this study was to evaluate whether DQ could prove useful to identify monosodium urate (MSU) and calcium pyrophosphate dehydrate (CPPD) crystals on permanent mounted stained slides. To this end, we studied 27 synovial fluid (SF) samples obtained from the knees of patients with the pseudogout (n=21) and acute gouty arthritis (n=6). Wet analysis for crystal detection and identification was performed within one hour of joint aspiration. In addition, we studied 16 inflammatory synovial effusions obtained from patients with knee arthritis not induced by crystals. For each SF, DQ stained slides were analyzed by 2 experienced doctors in SF analysis. The observers were blinded to the type of crystal present in the SF. Each slide was analyzed by compensated polarized and transmitted light microscopy. SF was considered positive if intracellular and/or extracellular crystals were clearly identified. In addition, the observers were asked to identify the type of the crystals using compensated polarized light microscopy. Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) of the DQ staining method were determined. 51 true positive and 28 true negative specimens were correctly classified (39 CPPD samples, 12 MSU samples, and 28 samples of crystals-unrelated arthropathies). All MSU specimens were correctly diagnosed

Polymetaphosphate based formulations for therapy of microcrystalline arthropaties

Soluble pharmaceutical composition for the treatment of articular pathologies comprising an effective amount of at least one linear or cyclic polymetaphosphate or a soluble and pharmaceutically acceptable salt thereof, and appropriate diluents

Axonal degeneration is mediated by necroptosis activation Necroptosis mediates axonal degeneration

Axonal degeneration contributes to functional impairment in several disorders of the nervous system, constituting an important target for neuroprotection. Several individual factors and subcellular events have been implicated in axonal degeneration, but the identification of an integrative signaling pathway activating this self-destructive process has remained elusive. Through pharmacological and genetic approaches, we tested whether necroptosis, a regulated cell death mechanism, implicated in the pathogenesis of several neurodegenerative diseases, is involved in axonal degeneration. Pharmacological inhibition of the necroptotic kinase RIPK1 using necrostatin-1 strongly delayed axonal degeneration in the peripheral and central nervous system of wild-type mice of either sex and protected in vitro sensory axons from degeneration after mechanical and toxic insults. These effects were also observed after genetic knock down of RIPK3, a second key regulator of necroptosis, and the downstream effector, MLKL RIPK1 inhibition prevented mitochondrial fragmentation in vitro and in vivo, a typical feature of necrotic death, and inhibition of mitochondrial fission by Mdivi also resulted in reduced axonal loss in damaged nerves. Furthermore, electrophysiological analysis demonstrated that inhibition of necroptosis delays not only the morphological degeneration of axons but also the loss of their electrophysiological function after nerve injury. Activation of the necroptotic pathway early during injury-induced axonal degeneration was evidenced by increased phosphorylation of the downstream effector MLKL. Our results demonstrate that axonal degeneration proceeds by necroptosis, defining a novel mechanistic framework in the axonal degenerative cascade for therapeutic interventions in a wide variety of conditions that lead to neuronal loss and functional impairment.SIGNIFICANCE STATEMENTWe show that axonal degeneration triggered by diverse stimuli is mediated by the activation of the necroptotic programmed cell death program by a cell-autonomous mechanism. We believe that this work represents a critical advance for the field since it identifies a defined degenerative pathway involved in axonal degeneration in both PNS and CNS, a process that has been proposed as an early event in several neurodegenerative conditions and a major contributor of neuronal death. The identification of necroptosis as a key mechanism for axonal degeneration, is an important step to develop novel therapeutic strategies for nervous system disorders, particularly those related to chemotherapy-induced peripheral neuropathies or CNS diseases in which axonal degeneration is a common factor

Ruptured abdominal aortic aneurysm in the elderly patient

The authors discuss several aspects of the management of ruptured abdominal aortic aneurysm in elderly patients. The cost-effectiveness and indications of repair of rAAA in elderly patients are analysed. A literature survey of risk-factors and results of open treatment of rAAA in elderly patients is made. The challenge of endovascular repair of rAAA in the elderly patient is discussed. Finally, the authors report their personal experience with AAA repair in 163 patients aged 75 years and older, operated on between January 2003 and September 2005(89 endoaneurysmal stent-grafts and 74 open repairs, 42 rAAA, 23 symptomatic AAA and 98 selective asymptomatic AAA)

Expression of macrophage migration inhibitory factor in diffuse systemic sclerosis

Objective: To evaluate whether, in patients with the diffuse form of systemic sclerosis (dSSc), macrophage migration inhibitory factor (MIF) production is dysregulated. Methods: 10 patients with dSSc and 10 healthy controls, matched for age and sex, were studied. MIF expression was evaluated by immunohistochemistry on formalin fixed skin biopsies of patients with dSSc and controls. MIF levels were assayed in the sera and in the supernatants of skin cultured fibroblasts by a colorimetric sandwich enzyme linked immunosorbent assay (ELISA). MIF concentrations in culture medium samples and in serum samples were compared by Student's two tailed t test for unpaired data. Results: Anti-MIF antibody immunostained the basal and mainly suprabasal keratinocytes. Small perivascular clusters of infiltrating mononuclear cells were positive; scattered spindle fibroblast-like cells were immunostained in superficial and deep dermal layers. The serum concentrations of MIF in patients with dSSc (mean (SD) 10705.6 (9311) pg/ml) were significantly higher than in controls (2157.5 (1288.6) pg/ml; p=0.011); MIF levels from dSSc fibroblast cultures (mean (SD) 1.74 (0.16) ng/2x10(5) cells) were also significantly higher than in controls (0.6 (0.2) ng/2x10(5) cells; p=0.008). Conclusion: These results suggest that MIF may be involved in the amplifying proinflammatory loop leading to scleroderma tissue remodelling

Ruptured abdominal aortic aneurysm in the elderly patient

The authors discuss several aspects of the management of ruptured abdominal aortic aneurysm in elderly patients. The cost-effectiveness and indications of repair of rAAA in elderly patients are analysed. A literature survey of risk-factors and results of open treatment of rAAA in elderly patients is made. The challenge of endovascular repair of rAAA in the elderly patient is discussed. Finally, the authors report their personal experience with AAA repair in 163 patients aged 75 years and older, operated on between January 2003 and September 2005(89 endoaneurysmal stent-grafts and 74 open repairs, 42 rAAA, 23 symptomatic AAA and 98 selective asymptomatic AAA)