Plasma acrolein level in rheumatoid arthritis increases independently of the disease characteristics.

OBJECTIVE: This study aimed to clarify whether plasma acrolein level actually increases in rheumatoid arthritis (RA) patients, and to elucidate whether any relationship exists between the levels and the RA background variables. METHODS: Plasma levels of protein-conjugated acrolein (PC-Acro) in 84 patients (RA group) and 298 normal individuals (Control group) were measured by enzyme-linked immunosorbent assay procedures. The data were statistically analyzed with Wilcoxon rank-sum test, multiple logistic regression analyses and Spearman's rank correlation coefficient. RESULTS: The RA group showed significantly higher PC-Acro levels than the Control group (median [interquartile range]: 80.5 [63.2-105.2] and 65.9 [58.9-78.1] nmol/ml, respectively). Of background factors giving influence to PC-Acro level in the combination of the two groups, 'diagnosis of RA positive' indicated strong correlation to high PC-Acro level (odds ratio: 2.96; 95% confidence interval: 1.54-5.71). These increases of PC-Acro in the RA patients did not correlate to their disease duration and/or inflammatory variables: PC-Acro level could elevate even in early RA patients showing negative inflammatory findings. CONCLUSION: Plasma levels of PC-Acro increased with RA, but the levels did not correlate with RA background variables. This report provides the basis for further studies of early diagnosis of RA as well as its pathogenesis.

Stereoselective Total Synthesis of Macrophage-Produced Prohealing 14,21-Dihydroxy Docosahexaenoic Acids.

Synthesis of 14S,21R- and 14S,21S-dihydroxy-DHA (diHDHA) among the four possible stereoisomers of 14,21-diHDHA was studied. Methyl (R)-lactate (>97% ee), selected as a C20-C22 fragment (DHA numbering), was converted to the C17-C22 phosphonium salt, which was subjected to a Wittig reaction with racemic C16-aldehyde of the C12-C16 part with the TMS and TBS-oxy groups at C12 and C14, yielding the C12-C22 derivative with 14R/S and 21R chirality. Kinetic resolution using Sharpless asymmetric epoxidation of the TBS-deprotected allylic alcohol with l-(+)-DIPT/Ti(O-i-Pr)4 afforded 14S-epoxy alcohol and 14R-allylic alcohol with >99% diastereomeric excess (de) for both. The CN group was introduced to the epoxy alcohol by reaction with Et2AlCN. The 14R-allylic alcohol was also converted to the nitrile via Mitsunobu inversion. Reduction of the nitrile with DIBAL afforded the key aldehyde corresponding to the C11-C22 moiety. The Wittig reaction of this aldehyde with a phosphonium salt of the remaining C1-C10 part followed by functional group manipulation gave 14S,21R-diHDHA. Similarly, ethyl (S)-lactate (>99% ee) was converted to 14S,21S-diHDHA. The chiral LC-UV-MS/MS analysis demonstrated that each of these two 14,21-diHDHAs synthesized using the presented total organic synthesis was highly stereoselective and identical to the macrophage-produced counterpart.

An interdigitated electrode with dense carbon nanotube forests on conductive supports for electrochemical biosensors.

A highly sensitive interdigitated electrode (IDE) with vertically aligned dense carbon nanotube forests directly grown on conductive supports was demonstrated by combining UV lithography and a low temperature chemical vapor deposition process (470 °C). The cyclic voltammetry (CV) measurements of K4[Fe(CN)6] showed that the redox current of the IDE with CNT forests (CNTF-IDE) reached the steady state much more quickly compared to that of conventional gold IDE (Au-IDE). The performance of the CNTF-IDE largely depended on the geometry of the electrodes (e.g. width and gap). With the optimum three-dimensional electrode structure, the anodic current was amplified by a factor of ∼18 and ∼67 in the CV and the chronoamperometry measurements, respectively. The collection efficiency, defined as the ratio of the cathodic current to the anodic current at steady state, was improved up to 97.3%. The selective detection of dopamine (DA) under the coexistence of l-ascorbic acid with high concentration (100 µM) was achieved with a linear range of 100 nM-100 µM, a sensitivity of 14.3 mA mol-1 L, and a limit of detection (LOD, S/N = 3) of 42 nM. Compared to the conventional carbon electrodes, the CNTF-IDE showed superior anti-fouling property, which is of significant importance for practical applications, with a negligible shift of the half-wave potential (ΔE1/2 < 1.4 mV) for repeated CV measurements of DA at high concentration (100 µM).

Normal sequential changes in neutrophil CD64 expression after total joint arthroplasty.

BACKGROUND: Neutrophil CD64 has been reported to be a sensitive and specific infection marker. Its measurement is thus considered to be useful in early diagnosis of post-operative periprosthetic infection. However, even its normal sequential changes after non-infectious total joint arthroplasty have remained ambiguous. Accordingly, we analyzed 2-week sequential neutrophil CD64 expression changes after total joint arthroplasty in order to clarify its normal postoperative kinetics. PATIENTS AND METHOD: From 41 patients who underwent primary total joint arthroplasties, peripheral blood samples were obtained at 1 day before (baseline) and 1, 3, 5, 7, and 14 days after surgery, and CD64 expression per cell was quantitatively measured. C-reactive protein (CRP) level, erythrocyte sedimentation rate (ESR) and white blood cell count (WBC) were simultaneously measured. RESULTS: No cases of postoperative local infection were observed. Levels of CD64 significantly elevated from day 1, peaked at day 3, and decreased significantly following day 5. Statistical analysis confirmed that significant differences existed between the baseline level and the levels at days 1 and 3, while no significant differences existed between the baseline level and those at days 5, 7 or 14. In 17 patients, CD64 peaked at over 2,000 molecules/cell, the level reported to be a cutoff value for distinguishing infection. Multiple regression analysis showed that the sole parameter of baseline CD64 level significantly explained the peak CD64 level. Postoperative CD64 peaks ranged from 1.6 to 2.7 times (median 1.9) the baseline levels. CRP, ESR and WBC also showed rapid elevations and all but WBC remained significantly higher than baseline at day 14. CONCLUSION: CD64 levels rise significantly, peaking within about 3 days following normal total joint arthroplasty, but decrease rapidly to near baseline within about 5 days. The data obtained can be expected to form a possible basis for early diagnosis of postoperative periprosthetic infection.

Oral and Intestinal Bacterial Substances Associated with Disease Activities in Patients with Rheumatoid Arthritis: A Cross-Sectional Clinical Study.

Intestinal bacterial compositions of rheumatoid arthritis (RA) patients have been reported to be different from those of healthy people. Dysbiosis, imbalance of the microbiota, is widely known to cause gut barrier damage, resulting in an influx of bacteria and their substances into host bloodstreams in animal studies. However, few studies have investigated the effect of bacterial substances on the pathophysiology of RA. In this study, eighty-seven active RA patients who had inadequate responses to conventional synthetic disease-modifying antirheumatic drugs or severe comorbidities were analyzed for correlations between many factors such as disease activities, disease biomarkers, intestinal bacterial counts, fecal and serum lipopolysaccharide (LPS), LPS-binding protein (LBP), endotoxin neutralizing capacity (ENC), and serum antibacterial substance IgG and IgA antibody levels by multiple regression analysis with consideration for demographic factors such as age, sex, smoking, and methotrexate treatment. Serum LBP levels, fecal LPS levels, total bacteria counts, serum anti-LPS from Porphyromonas gingivalis (Pg-LPS) IgG antibody levels, and serum anti-Pg-LPS IgA antibody levels were selected for multiple regression analysis using Spearman's correlation analysis. Serum LBP levels were correlated with disease biomarker levels, such as erythrocyte sedimentation rate (p < 0.001), C-reactive protein (p < 0.001), matrix metalloproteinase-3 (p < 0.001), and IL-6 (p = 0.001), and were inversely correlated with hemoglobin (p = 0.005). Anti-Pg-LPS IgG antibody levels were inversely correlated with activity indices such as patient global assessments using visual analogue scale (VAS) (p = 0.002) and painVAS (p < 0.001). Total bacteria counts were correlated with ENC (p < 0.001), and inversely correlated with serum LPS (p < 0.001) and anti-Pg-LPS IgA antibody levels (p < 0.001). These results suggest that substances from oral and gut microbiota may influence disease activity in RA patients.

Efficacy and safety of single-dose mizoribine for patients with rheumatoid arthritis: results at 6 months after switching from a multiple-dose regimen without a change in total daily dose.

To determine the efficacy and safety of single-dose mizoribine (MZR) for patients with rheumatoid arthritis (RA), a 6-month, single-arm, open-label, prospective observation study was performed. In patients who had been taking MZR at 100-150 mg/day in 2-3 divided portions continuously for at least 3 months, and who had shown a lack of clinical response, or escape (defined as a lack of response at the time of switching, even if some form of response had been shown before that), multiple-dose administration was switched to single-dose administration without changing the total daily dose. Efficacy was assessed in terms of the disease activity score, using the 28-joint count and erythrocyte sedimentation rate (DAS 28-ESR). Of the 34 enrolled patients, 28 met all the eligibility criteria and were assessed for efficacy, and finally 26 patients were able to receive the single-dose regimen throughout the full 6 months. The DAS28-ESR showed a significant decrease from 2 months after switching, and 46.4% of the 28 patients finally achieved a good or moderate response (3 and 10 patients, respectively). With regard to safety, no serious adverse events were observed. In conclusion, the administration of MZR at 100 or 150 mg in a single dose is thought to be a useful alternative form of MZR therapy.

The usefulness of neutrophil CD64 expression in the diagnosis of local infection in patients with rheumatoid arthritis in daily practice.

BACKGROUND: The diagnosis of local infection in patients with rheumatoid arthritis (RA) is frequently difficult because clinical signs and symptoms and laboratory test results of local infection are also observed in arthritis of active RA. The need for a specific marker of infection is high in RA patients. The usefulness of neutrophil CD64 expression (CD64) to diagnose local musculoskeletal infection (local infection) and discriminate local infection from RA-related inflammation in RA patients was examined. METHODS: CD64 was measured by a quantitative method using flow cytometry in 61 RA patients in whom local infection was suspected, and the usefulness of CD64 was examined by comparing the findings with clinical results. RESULTS: There were 25 patients with local infection and 36 patients without infection. The median CD64 value the patients with local infection was 3148 molecules/cell (interquartile range [IQR], 2140-6231) and that of the patients without infection was 1106 molecules/cell (IQR, 804-1464) with a statistically significant difference (P < 0.0001). In contrast, no significant difference between the groups was observed in C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and white blood cell (WBC) count. The area under the curve of CD64 calculated by receiver operating characteristic curve analysis was larger than that of CRP, ESR, or WBC count, suggesting that CD64 has superior ability to discriminate of infection compared to these other markers. When the cutoff value of CD64 was set at 2000 molecules/cell, the sensitivity and specificity of CD64 for the detection of local infection in RA patients were 76.0% and 94.4%, respectively. CONCLUSIONS: CD64 is a useful marker in RA patients to discriminate local infection from RA-related inflammation.

Neutrophil CD64 expression as a diagnostic marker for local infection and crystal-induced arthritis.

The usefulness of neutrophil CD64 expression was examined in diagnosing local infection, including soft tissue, bone, and joint infections. Among 99 patients suspected of local infection, 31 were confirmed and 68 patients were not. The CD64 level of patients with local infection was significantly higher than in those without infection [4,193 ± 1,132 vs. 1,017 ± 59 molecules/cell (mean ± standard deviation); p < 0.001]. The area under the curve of CD64 calculated by receiver operating characteristic curve analysis was larger than that of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), or white blood cell (WBC) count. In addition, CD64 levels of patients with crystal-induced arthritis remained within cutoff value (2,000 molecules/cell). These data suggest that measuring CD64 expression can be a useful diagnostic marker for local musculoskeletal infection and crystal-induced arthritis.

Rheumatoid arthritis associated with osteopetrosis.

Osteopetrosis is an inherited disorder characterized by reduced bone resorption. We here report a rare case of osteopetrosis associated with rheumatoid arthritis. The patient was diagnosed as autosomal dominant osteopetrosis type II in his youth and developed rheumatoid arthritis at 42 years of age. In spite of the severe inflammation and rapid progression of cartilage destruction, the progression of bone erosion was slow in this patient.

Prevalence of joint replacement surgery in rheumatoid arthritis patients: cross-sectional analysis in a large observational cohort in Japan.

The purpose of this study was to describe the prevalence of total joint arthroplasty (TJA) in Japanese rheumatoid arthritis (RA) patients undergoing conventional drug treatment in a large observational cohort in Japan. A total of 5,177 RA patients were studied for the prevalence of TJA, who were enrolled in the NinJa database during the fiscal year of 2006. The cases of 2,695 RA patients with more than ten years of disease duration were extracted and subjected to further analysis. The prevalence of TJA increased in accordance with the disease duration, and the prevalence was markedly increased after ten years. Among the 2,695 patients with more than ten years of disease duration, 1,431 TJAs were performed in 645 (24.6%) patients. The patients with TJA had higher disease activity than those without TJA. In this cross-sectional study, TJAs were performed in approximately a quarter of the Japanese RA patients with more than ten years of disease duration. The result showed that patients with higher disease activity required TJA.

No changes in infliximab levels in blood stored for preoperative autologous blood donation.

Rheumatoid arthritis (RA) patients requiring total joint arthroplasties under administration of infliximab, which may remain in donated blood if preoperative autologous blood donation (PABD) is undertaken for the surgery, may risk infection. We clarified infliximab hemokinetics in blood stored for such patients. A 20-ml blood sample was obtained from each of the ten RA patients receiving infliximab at just after administration and at 2 and 4 weeks following the administration of infliximab, mixed with 2.8 ml citrate-phosphate-dextrose-adenine (CPDA-1) and stored at 4-6 degrees C. Plasma levels of infliximab in the stored blood were measured just after-mixture with CPDA-1, and at 2 and 4 weeks following the start of storage. Serum levels were also measured just before infliximab administration and at each phlebotomy. The plasma infliximab levels in the stored blood remained close to their original serum levels at the time of each corresponding phlebotomy, only somewhat influenced by dilution of CPDA-1, and sustained for 4 weeks following the start of storage, unlike in vivo, where levels decreased. This suggests that in order to prevent side effects, the later after infusion of infliximab the phlebotomy occurs, the better, and that the amount of stored blood transfusion should be consistent with that of blood loss.

Measurement of articular cartilage volumes in the normal knee by magnetic resonance imaging: can cartilage volumes be estimated from physical characteristics?

In recent times several studies have been performed on magnetic resonance imaging (MRI) sequences for imaging cartilage. A fat-suppressed three-dimensional sequence is one such noteworthy example. More recent studies have reported that the total volume of cartilage in a knee joint can be elucidated using this sequence. Based on these studies, we hypothesized that the total volume of cartilage in the knee joint may reflect certain other physical characteristics. The purpose of the current study was to clarify the articular cartilage volumes of the patella and femur in the human knee joints of healthy adults using MRI and to analyze the correlation of these volumes with other physical characteristics. The material comprised 68 knees of 68 Japanese healthy volunteers, aged from their twenties to their forties (37 men and 31 women) who had no past history of joint disease or trauma in the legs. The knees were imaged by MRI with a fat-suppressed three-dimensional sequence, and the cartilage volumes were calculated by computer processing. The factors analyzed were age, body weight, height, leg length, foot size, circumferences of the thigh and lower leg, the distance between medial and lateral femoral condyles, the diameter of the tibial head, body mass index, general joint laxity, quadriceps angle, and leg-heel alignment. The mean cartilage volume was 7.6 +/- 1.6 cm(3) (8.3 +/- 1.6 cm(3) in men, 6.7 +/- 0.9 cm(3) in women). It was significantly larger in men than in women. However, the volume positively correlated with body weight, height, leg length, and foot size, without distinction of gender or age. Based on these data, a multiple regression analysis was developed: cartilage volume = 0.113 x height - 11.053. We concluded that the cartilage volume depends on physical size regardless of gender, and it can be estimated from factors of physical size.