DSC measurement of cartilage destruction caused by septic arthritis

Summary Treatment of a bacterial arthritis is a challenging task for a clinician as inadequate therapy can cause cartilage destruction and can result in severe osteoarthritis of the affected joint. The development of cartilage destruction in septic arthritis is not known in details. The aim of this study was to follow this process by calorimetric method. We induced experimental septic arthritis in knee joints of seven New Zealand rabbits by single inocculation of Staphylococcus aureusOKI 112001 culture (1.5 mL 8·10⁸±5% c.f.u.). The first rabbit died on the 11^thday. At that time all the other subjects were made overslept and samples were isolated from the cartilage of the femurs for calorimetric measurement. The DSC scans clearly demonstrated the development of infective structural destruction in cartilage from the first to the tenth day of incubation. In case of healthy control the melting temperatures (T_m) were: 49.7, 55 and 63.4°C and the total calorimetric enthalpy change (ΔH) was 0.55 J g^-1. After the first day the enthalpy decreased (0.375 J g^-1), the first two transition temperature shifted towards higher temperature: 57 and 63.15°C. Up to the fourth day the effect of infection culminated with T_mof 49.3, 55.9, 59.4, 62.8°C and further decrease of the ΔH. At the fifth day the effect of infection is culminated in two separable thermal denaturation events (with 55 and 63.3°C T_ms) with high jump in ΔHindicating the dramatic change of the structure of rabbit cartilage, so this time elapsed seems to be critical from the point of view of practical clinical relevance too. Between the 7^thand 11^thdays practically we had same melting temperatures (50 and 63°C) with low (~0.24 J g^-1) enthalpy.     

Diffusion-weighted MR imaging for assessing synovitis of wrist and hand in patients with rheumatoid arthritis: A feasibility study

The purpose of this study was to investigate the feasibility of diffusion-weighted imaging (DWI) in detecting synovitis of wrist and hand in patients with rheumatoid arthritis (RA) and evaluate its sensitivity, specificity and accuracy as compared to T₂-weighted imaging (T2WI) with short tau inversion recovery (STIR) with the reference standard contrast-enhanced magnetic resonance imaging (CE-MRI). Twenty-five patients with RA underwent MR examinations including DWI, T2WI with STIR and CE-MRI. MR images were reviewed for the presence and location of synovitis of wrist and hand. The sensitivity, specificity and accuracy of DWI and T2WI with STIR were calculated respectively and then compared. All patients included in this study completed MR examinations and yielded diagnostic image quality of DWI. For individual joint, there was good to excellent inter-observer agreement (k = 0.62–0.83) using DWI images, T2WI with STIR images and CE-MR images, respectively. There was a significance between DWI and T2WI with STIR in analyzing proximal interphalangeal joints II–V, respectively (P < 0.05). The k-values for the detection of synovitis indicated excellent overall inter-observer agreements using DWI images (k = 0.86), T2WI with STIR images (k = 0.85) and CE-MR images (k = 0.91), respectively. Overall, DWI demonstrated a sensitivity, specificity and accuracy of 75.6%, 89.3% and 84.6%, respectively, for detection of synovitis, while 43.0%, 95.7% and 77.6% for T2WI with STIR, respectively. DWI showed positive lesions much better and more than T2WI with STIR. Our results indicate that DWI presents a novel non-invasive approach to contrast-free imaging of synovitis. It may play a role as an addition to standard protocols.     

UHPLC–MS/MS analysis of sphingosine 1‐phosphate in joint cavity dialysate and hemodialysis solution of adjuvant arthritis rats: Application to geniposide pharmacodynamic study

Geniposide (GE) is an iridoid glycoside compound with anti‐inflammatory effect. The potential of sphingosine 1‐phosphate (S1P) as a plasma marker in human diseases was suggested recently in the literature, which demonstrated that, in patients with inflammatory diseases, plasma S1P was elevated. It follows that the obstructive coronary artery disease can be predicted with serum S1P. Therefore, S1P can also be potentially used as a pharmacodynamic marker to study adjuvant arthritis (AA) rats. In the current study, a UHPLC–MS/MS method combined with the microdialysis sampling technique (using FTY720 phosphate as an internal standard) was adopted and validated to measure S1P levels in the hemodialysis fluid and joint cavity dialysates of AA rats after oral administration of GE. A S1P concentration–time curve in the dialysate was established in this study. It was demonstrated that GE exerted an anti‐inflammatory effect by reducing AA‐induced elevated S1P levels. It is showed that changes in S1P concentrations over time can be used to monitor the pharmacodynamic effects of GE in treating AA rats in pharmacodynamic studies.     

The Physical,Chemical and Biological Effects by Room Temperature Ceramic Far‐infrared Ray Emitting Material Irradiated Water: A Pilot Study

Far infrared ray (FIR) is non‐ionizing electromagnetic radiation with wavelengths of 4‐16 μm. Ceramic far infrared ray emitting materials (cFIR) are sources of FIR that exhibit only non‐thermal effects at room temperature. Certain physical, chemical and biological effects of cFIR irradiation were investigated in this study that heretofore has not been well characterized. We demonstrated that cFIR irradiation reduced the size of water clusters, and significantly increased the freezing temperature of water. We also observed an increase in the volatility of a complex mixture of alcohol, water and solutes and recorded elevated total phenol in green tea infusions prepared using cFIR irradiated water. The effects of cFIR irradiated water on living cells were also investigated. The MC3T3‐E1 murine osteoblast cells grown in the presence of cFIR irradiated water exhibited anti‐oxidative effects on H₂O₂‐mediated toxicity, as evidenced by late stage increase in intracellular alkaline phosphatase. The presence of cFIR irradiated water also resulted in significant decrease in COX‐2 production in the chondrosarcoma cell line, SW1353, in response to lipopolysaccharide induction. Based on our findings, the possible anti‐inflammatory effects of cFIR irradiated water and implications of our study with regard to bone and joint health were discussed.     

Synthesis and Crystal Structure of (7R,8S,9S,12S)-13,14-dehydro-1-(4'-fluorobenzyloxy)-N-methyl-2,13-dimethoxy-12-hydroxymorphinane

The title compound (7R,8S,9S,12S)-13,14-dehydro-1-(4-fluorobenzyloxy)-N-methyl-2,13-dimethoxy-12-hydroxymorphinane was synthesized by the reaction of O-4-bromobenzyl-sinomenine with lithium aluminum hydride.Its chemical structure was determined by X-ray single-crystal diffraction.The crystal belongs to triclinic,space group P1 with M r=439.51,a=7.7236(1),b=8.7282(1),c=9.9342(2),α=81.176(1),β=67.43,γ=64.577(1)°,μ=0.76 mm-1,V=558.45(2)3,Z=1,D c=1.307 mg/m 3,F(000)=234 and T=133 K.     

Detection of AMP‐activated protein kinase α1 by isoelectric focusing in sera of patients with metabolic diseases

The aim of this study is to determine by isoelectric focusing the level of AMPKα1, an energy sensor, in sera of patients who are in energy‐demanding situation. After Western blotting, detection was performed with specific antibodies against AMPKα and its phosphorylated form. To evaluate the effect of weight loss on AMPK, sera from 24 patients were collected before and after intragastric balloon insertion over a 16‐week follow‐up period. Compared to baseline, all patients showed postoperatively an increase of AMPK. Patients with ischemic heart, with inflammatory bowel disease, with chronic undernutrition or with hepatic diseases were examined. Compared to control subjects, the majority of them showed a significant increase of AMPK. These results suggest that serum AMPK may have a potential for diagnosis of several metabolic diseases. However, this has to be confirmed by further studies with additional biomarkers and with more specific techniques.     

The association between the CASP5 rs7939842 polymorphism and the risk of rheumatoid arthritis in Chinese Han individuals

BackgroundThe association between inflammatory cysteinyl aspartate protease-5 (CASP5) and the susceptibility to rheumatoid arthritis (RA) remains unclear. This study examined whether the CASP5 rs7939842 polymorphism affects RA risk in Chinese Han individuals.MethodsThis study recruited 805 RA patients and 1095 healthy controls to investigate the association between the CASP5 rs7939842 polymorphism and RA risk. Genotype was examined using the 48-Plex SNPscan? Kit. Plasma CASP5 levels were determined using enzyme-linked immunosorbent assays, and CASP5 gene expression was detected by quantitative polymerase chain reaction in 40 RA patients and 40 healthy controls.ResultThe CASP5 rs7939842 polymorphism G allele is a putative risk factor for RA. After stratified analyses, this polymorphism increased the risk of RA among CRP-, ACPA-, RF-, and ESR-positive individuals, as well as individuals with DAS28 ≥ 3.20 and functional class III + IV. Furthermore, the plasma CASP5 levels and CASP5 mRNA expression were higher in RA patients than in healthy controls.ConclusionThe CASP5 rs7939842 polymorphism appears to be associated with an elevated risk of RA in Chinese Han individuals. Blood CASP5 protein and mRNA levels were significantly higher in RA patients than in healthy controls.     

以琼脂凝胶固定热聚IgG制备类风湿关节炎免疫吸附剂的研究

以环氧氯丙烷对琼脂凝胶珠进行活化反应后键联热聚ＩｇＧ,制成一种新型类风湿关节炎免疫吸附剂。确定了最佳制备条件,使凝胶上环氧基的含量达１１０μｍｏｌ／ｇ,对热苯ＩｇＧ的固定量达６ｍｇ／ｇ。在体外条件下吸附剂对三种类风湿因子ＩｇＭＲＦ,ＩｇＧＲＦ及ＩｇＡＲＦ的吸附量分别达３４００,２２５０和２４００ＩＵ／ｇ,具有良好的应用前景。     

Functional Analysis of Autoantibody Signatures in Rheumatoid Arthritis

For the identification of antigenic protein biomarkers for rheumatoid arthritis (RA), we conducted IgG profiling on high density protein microarrays. Plasma IgG of 96 human samples (healthy controls, osteoarthritis, seropositive and seronegative RA, n = 24 each) and time-series plasma of a pristane-induced arthritis (PIA) rat model (n = 24 total) were probed on AIT’s 16k protein microarray. To investigate the analogy of underlying disease pathways, differential reactivity analysis was conducted. A total of n = 602 differentially reactive antigens (DIRAGs) at a significance cutoff of p < 0.05 were identified between seropositive and seronegative RA for the human samples. Correlation with the clinical disease activity index revealed an inverse correlation of antibodies against self-proteins found in pathways relevant for antigen presentation and immune regulation. The PIA model showed n = 1291 significant DIRAGs within acute disease. Significant DIRAGs for (I) seropositive, (II) seronegative and (III) PIA were subjected to the Reactome pathway browser which also revealed pathways relevant for antigen presentation and immune regulation; of these, seven overlapping pathways had high significance. We therefore conclude that the PIA model reflects the biological similarities of the disease pathogenesis. Our data show that protein array analysis can elucidate biological differences and pathways relevant in disease as well be a useful additional layer of omics information.