The utility of alizarin red s staining in calcium pyrophosphate dihydrate crystal deposition disease

OBJECTIVE: To determine the most suitable staining method for preservation and detection of calcium pyrophosphate dihydrate (CPPD) crystals in histological sections of patients with CPPD crystal deposition disease. METHODS: Paraffin sections of CPPD crystal-bearing tissues of 31 patients were stained with hematoxylin and eosin (H&E) and Alizarin red S (ARS). For H&E, the sections were treated with Mayer's hematoxylin (pH 2.3) for 5 min and with eosin alcohol (pH 4.1) for 1 min. For ARS, 1% ARS dissolved in distilled water was adjusted to pH 6.4 by adding 0.1% ammonia solution drop by drop while stirring. As controls, unstained sections were soaked in 1% citric acid monohydrate solution (CAMS, pH 2.3) for 5 or 10 min. The histological preparations were examined under a compensated polarized light using a first-order red compensator. We counted the number of weakly positive birefringent CPPD crystals in 3 high power fields (HPF, 0.272 mm2). RESULTS: CPPD crystals were seen clearly in most specimens stained with ARS, but were markedly reduced in tissue sections stained with H&E or CAMS. The number of CPPD crystals detected in sections stained by ARS (1723 +/- 683 per 3 HPF, mean +/- standard deviation) was significantly higher compared with H&E, CAMS (5 min), and CAMS (10 min) (401 +/- 374, 1022 +/- 616, and 494 +/- 636 per 3 HPF, respectively; p < 0.001, each). CONCLUSION: Standard H&E staining reduces the number of visible CPPD crystals, probably due to the strong acidity of both hematoxylin and eosin solutions, whereas the ARS stain seems to preserve a large number of CPPD crystals. The utility of ARS staining may improve the identification of CPPD crystals and contribute to a correct diagnosis of CPPD crystal deposition.     

The neurotoxicity of psychoactive phenethylamines “2C series” in cultured monoaminergic neuronal cell lines

Forensic Toxicology - The aim of this study was to evaluate the neurotoxicity of psychoactive abused 2,5-dimethoxy-substituted phenethylamines “2C series” in monoaminergic neurons....     

Terminal deletion of the long arm of chromosome 3 [46,XX,del(3)(q27→qter)]

We report on a terminal deletion of the long arm of chromosome 3 [46,XX,del(3)(q27→qter)] in a female newborn infant who died 45 hours after delivery and had multiple congenital abnormalities including bilateral anophthalmia, congenital heart disease, and abnormal genitalia. The findings are compared to those of four previously reported cases with terminal del (3q). © 1996 Wiley-Liss, Inc.     

Definite familial multiple system atrophy with unknown genetics

Multiple system atrophy (MSA) is an oligodendrogliopathy of presumably sporadic origin, characterized by prominent α‐synuclein inclusions with neuronal multisystem degeneration, although a few Mendelian pedigrees have been reported. Here we report two familial cases of MSA of unknown genetic background. One patient was diagnosed as a possible MSA‐C (cerebellar dysfuntion) case, and the other as clinically possible MSA‐P (parkinsonism), which turned out to be definite MSA, based on a detailed autopsy. The neuropathology showed extensive deposition of α‐synuclein in the glia as well as in the neurons located in the cerebral cortices and hippocampal systems, although neither multiplication of the SNCA gene or mutations in COQ2 gene were identified in the family concerned.     

Serum soluble interleukin‐2 receptor level is more sensitive than angiotensin‐converting enzyme or lysozyme for diagnosis of sarcoidosis and may be a marker of multiple organ involvement

Skin lesions in sarcoidosis are often the initial symptoms that enable the dermatologist to be the first to diagnose this granulomatosis. However, diagnosis is sometimes very problematic. In 2015, the diagnostic criteria for sarcoidosis were updated in Japan, with elevated serum soluble interleukin‐2 receptor (sIL‐2R) replacing negative tuberculin reaction. Therefore, we assessed the clinical utility of sIL‐2R compared with two other common markers, angiotensin‐converting enzyme (ACE) and lysozyme, in patients who visited the dermatology clinic. Data from 72 patients showed that sIL‐2R was more sensitive than both ACE and lysozyme in supporting a diagnosis of sarcoidosis (52.8%) compared with ACE (29%) and lysozyme (26.4%). Additionally, the sIL‐2R level was significantly higher in patients with multiple organ involvement and parenchymal infiltration. Patients with elevated sIL‐2R levels had higher serum ACE and lysozyme levels, a higher incidence of pulmonary involvement, more severe chest radiographic stage and a high incidence of expression‐specific signs by imaging analysis. Receiver–operator curve analysis showed that sIL‐2R was a better marker at the threshold cut‐off point compared with ACE and lysozyme for identifying patients with multiple organ involvement, detecting patients with pulmonary disease and parenchymal infiltration as well as predicting the presence of specific signs in the diagnosis of sarcoidosis. Moreover, the kinetics of sIL‐2R levels correlated closely with clinical manifestations, in contrast to the modest changes of ACE and lysozyme levels during the follow‐up period. In conclusion, sIL‐2R may be considered a good marker for diagnosis and a potential indicator of disease activity.     

Pharmacodynamic monitoring of calcineurin phosphatase activity in transplant patients treated with calcineurin inhibitors

Calcineurin inhibitors, tacrolimus and cyclosporine, have been widely used to prevent the rejection or graft-versus-host disease after transplantations. Since these drugs have a narrow therapeutic range and show large inter- and intraindividual pharmacokinetic variability, frequent therapeutic drug monitoring is required to control their blood concentrations. Even with blood concentrations within the therapeutic range, some patients still experience acute rejection or infections. Tacrolimus and cyclosporine form a complex with their respective binding proteins, immunophilins, which in turn inhibit the phosphatase activity of calcineurin, a key enzyme in the activation of T lymphocytes. Pharmacodynamic assessment of calcineurin phosphatase activity in combination with the monitoring of blood concentrations has been studied. The inhibitory effects on calcineurin activity in peripheral blood mononuclear cells differed between tacrolimus and cyclosporine in transplant patients. The pharmacodynamics of both drugs shows great inter- as well as intraindividual variation, and acute rejection was associated with calcineurin activity. Calcineurin activity at trough time points was suggested as a single surrogate predictor for overall calcineurin activity throughout dosing periods. Monitoring of calcineurin phosphatase activity might be useful to determine the therapeutic range of tacrolimus and cyclosporine concentrations for an individual patient treated with a calcineurin inhibitor.     

Confocal immunolocalization of VE-cadherin- and CXC chemokine-expressing endothelial cells in periapical granulomas

Aim To determine whether endothelial cells (ECs) in periapical granulomas can express vascular endothelial (VE)‐cadherin, CXCL8 and CXCL10 by examining with two‐colour confocal laser scanning microscope. Methodology Periapical lesions were surgically removed from patients with chronic periapical periodontitis (n = 20), and the paraffin‐embedded sections were prepared after being fixed with cold acetone. The 7‐μm‐thick sections were stained with haematoxylin–eosin and then examined pathologically using a light microscope. The lesions diagnosed as periapical granulomas (17 specimens) were analysed further using immunofluorescence and antibodies specific for human VE‐cadherin, CXCL8, and CXCL10. The slides were carefully examined using a confocal laser scanning microscope. The numbers of positive ECs were counted, and the comparison between VE‐cadherin‐positive ECs and CXCL8 or CXCL10 was assessed statistically using one‐way ANOVA followed by a Student–Newman–Keuls test. Results The expression of CXCL8 and CXCL10 by ECs was detected in 60.4 ± 13.4 and 67.2 ± 13.9%, respectively. However, the percentage of VE‐cadherin‐expressing ECs was 40.4 ± 10.5%, which was significantly lower (P < 0.01) than CXCL8 and CXCL10‐expressing ECs. Two‐colour immunofluorescence staining revealed that ECs co‐expressed VE‐cadherin and CXCL8 (37.4 ± 14.1%) or CXCL10 (39.1 ± 13.8%). Conclusions VE‐cadherin expression in ECs was lower than CXCL8 and CXCL10, suggesting that inflamed ECs in periapical granulomas could increase vascular permeability and that leukocyte chemotaxis mediated by ECs might occur. These findings may suggest the possibility that ECs could play a pivotal role in cell recruitment in periapical granulomas.