Liquid chromatography-tandem mass spectrometry method for the determination of anthelmintics in alfalfa plants.

A simple and inexpensive liquid chromatography-tandem mass spectrometric method for the determination of anthelmintics in alfalfa plants (Medicago sativa L.) was developed and validated. Anthelmintics in plant leaves and stems (green chops) were extracted with methanol/acetonitrile (7:3, v/v) followed by a concentration and clean-up step using solid-phase extraction (Strata-X, 500 mg, 6 ml cartridge). After drying with nitrogen gas, the adsorbed analytes were eluted with methanol/acetonitrile (50:50, v/v) mixture followed by 100% acetonitrile. Chromatographic separation was achieved on an Atlantis T-3 (2.1 × 100 mm × 3 µm) analytical column with a Phenomenex guard cartridge (C8, 4 × 3 mm) attached to a Waters triple quadrupole mass spectrometer operated in positive electrospray ionisation mode with selected reaction monitoring. Samples were analysed using gradient elution at a flow rate of 0.35 ml min⁻¹. The mobile phase consisted of a 10 mM ammonium formate solution in (A) water/acetonitrile (90:10, v/v) and (B) methanol/acetonitrile (50:50, v/v). The method was validated for levamisole, fenbendazole, fenbendazole sulphoxide and fenbendazole sulphone at 10, 20 and 40 µg kg⁻¹ and for eprinomectin at 20, 40 and 80 µg kg⁻¹. Limits of quantification (LOQ) were 10 µg kg⁻¹ for all analytes except eprinomectin, which had an LOQ of 20 µg kg⁻¹. The overall mean recovery in green plants was between 74.2% and 81.4% with repeatabilities ranging from 2.2% to 19.1% and reproducibilities in the range 3.8-8.7%. The validated method was applied to plant samples in a study on the behaviour of anthelmintic drugs in a soil, plant and water system.

Increased serum levels of cartilage oligomeric matrix protein in patients with psoriasis vulgaris: a marker for unknown peripheral joint involvement?

OBJECTIVE: Cartilage oligomeric matrix protein (COMP) is a parameter for the current extent of cartilage destruction. It has been shown that the release pattern of cartilage oligomeric matrix protein in serum reflects cartilage turnover. The aim of our study was to explore the utility of sCOMP as a marker for disease activity in patients with active psoriatic arthritis (PsA) in comparison to a control group only with psoriasis vulgaris (PV). METHODS: Serum levels of COMP were measured in 64 patients with PsA and psoriasis vulgaris. The control group consisted of a population with PV from a dermatological outpatient clinic. ELISA-tests were used to detect sCOMP levels according to the manufacturer instructions. RESULTS: In our 64 patients with PsA, we found increased sCOMP levels, which correlated significantly with inflammatory parameters and the number of swollen joints. Patients with active PsA had significantly higher sCOMP levels (p<0.0001) than the 39 patients with a low inflammatory status. In our control group with PV we also found elevated sCOMP levels, which correlated significantly with the increased C-reactive protein (CRP) levels in this group. The difference between the PsA and the PV group was not significant (p=0.092). CONCLUSION: In our study, sCOMP has been demonstrated to be an indicator for disease activity in patients with PsA. Patients with active PsA showed significantly elevated sCOMP levels compared to the patients with low clinical and laboratory disease activity. The increased sCOMP levels in our control group with PV indicate that all patients with psoriatic lesions should be screened for additional joint involvement and should lead to an exact joint examination.

Serum levels of cartilage oligomeric matrix protein (COMP): a rapid decrease in patients with active rheumatoid arthritis undergoing intravenous steroid treatment.

To examine the influence of intravenous steroid-treatment (IST) on serum levels of Cartilage oligomeric matrix protein (COMP) in patients with active rheumatoid arthritis (RA). Serum levels of COMP and C-reactive protein (CRP) were measured in 12 patients with highly active RA (Steinbrocker stages II-IV) and in 5 patients with highly active reactive arthritis (ReA) (positive testing for HLA-B27) before starting daily IST. Patients received a total steroid dosage between 100 and 500 mg of prednisolone. COMP was measured by a commercially available sandwich-type ELISA-kit developed by AnaMar Medical AB, Sweden. Statistical evaluation was calculated by paired t test. In the RA group, COMP levels ranged from 6.3 to 19.4 U/l (mean 12.9 U/l), CRP from 5 to 195 mg/l (mean 77.8 mg/l), the COMP levels of the ReA group ranged from 5.1 to 7.4 U/l (mean 7.9 U/l), the CRP levels from 13 to 126 mg/l (mean 49 mg/l). We found a significant difference between the initial COMP levels in RA+ and ReA patients (P<0.005). In contrast to the ReA group, serum-COMP levels of RA+ patients (P<0.004) and the VAS (P<0.0001) decreased significantly within 2-10 days after the first treatment with steroids. The CRP levels remained unchanged in both groups. Our results indicate that the intravenous treatment with steroids in patients with highly active RA leads to a significant decrease of cartilage degradation. COMP seems to be a valuable parameter not even as a prognostic factor, but as a marker for monitoring the therapy response in patients with RA.

Osteoprotegerin and the receptor activator of NF-kappa B ligand in the serum and synovial fluid. A comparison of patients with longstanding rheumatoid arthritis and osteoarthritis.

We examined OPG and soluble RANKL in the serum (sOPG, sRANKL) and synovial fluid (synOPG, synRANKL) in patients with rheumatoid arthritis (RA) and osteoarthritis (OA). OPG and RANKL were measured in 85 patients (44 with RA, 41 patients with OA) in serum and synovial fluid as well. For measuring of OPG and RANKL ELISA tests were used. The results of OPG and RANKL were compared with clinical and radiological scores. We found a negative correlation for OPG and RANKL in synovial fluids: not only for the whole group of patients (P < 0.003, r = -0.32), but also for the subgroups (RA: P < 0.04, r = -0.28, OA: P < 0.002, r = -0.54). SRANKL and synRANKL were positively correlated in the whole group (P < 0.01, r = 0.25) and in the OA group (P < 0.02, r = 0.35); the RA group was showing a trend (P < 0.063, r = 0.24), however. Serum OPG was lower in RA, synOPG higher in OA. The difference between the two patient groups was only significant for synOPG (P < 0.03, r = 0.056), but not for sOPG (P < 0.09, r = 0.19), sRANKL (P < 0.43, r = 0.85) or synRANKL (P < 0.11, r = 0.22). The synOPG:synRANKL ratio was significantly correlated with the Larsen score (P < 0.004, r = 0.38). Synovial OPG is significantly decreased in rheumatoid joints, whereby synovial RANKL is increased. Lower synOPG could reflect a lower protective effect on bone, thus leading to an earlier and more pronounced bone destruction in RA. However, the effect of different mediators for joint destruction in RA and OA seems not to be important to the pathophysiological changes in the joints. The upregulation of serum OPG might be the result of the inflammation; in contrast, an upregulation of RANKL could not be found in the serum of patients with RA and OA.

Influence of molecular structure on sorption of phenoxyalkanoic herbicides on soil and its particle size fractions.

The sorption and desorption behaviors of four phenoxyalkanoic acid herbicides and their metabolites on four agricultural soils and soil particle size fractions were examined. Generally, there was a trend of increasing adsorption and decreasing desorption in the order mecoprop < MCPA < dichlorprop < 2,4-D. The significant increase in adsorption of the phenolic metabolites can be explained by their lower polarity and enhanced partition in the organic soil matrix. Estimation of sorption distribution coefficients from particle size fraction adsorption data was possible for a sandy soil and a silty Cambisol soil only. It is suggested that increasing steric demand, for example, molecular volume, and slight changes in the polarity of the compounds affect their adsorption properties. Comparison of adsorption and desorption data of structurally similar compounds obtained from a variety of soils allows investigation of structure-induced differences in sorption strength.

[Serum soluble CD44 isoform variant 5 level in patients with seropositive rheumatoid arthritis treated with cyclosporin A]. / Serumspiegel löslicher CD44-Isoform-Variante 5 von Patienten mit seropositiver Rheumatoid-Arthritis unter Cyclosporin-A-Therapie.

CD44 is a widely expressed cell surface glycoprotein which is involved in many cell-cell and cell-matrix interactions. Expression of soluble CD44 splice variants is strictly regulated and is linked to a high rate of cell division. Serum levels of soluble CD44 variant 5 (sCD44v5) were determined in 14 patients with erosive RA. Patients were divided into two groups. In group 1 cyclosporin A treatment (CYA) was initiated after the first visit. In group 2 preliminary CYA was continued. Controls were performed after 6 months. We found a significant decrease of swollen joint count (SJC) and sCD44v5 in group 1. No effect of CYA was found on c-reactive protein, erythrocyte sedimentation rate and IgM-rheumatoid factor (IgM-RF). In group 2 a significant decrease of CRP was found. Therefore we conclude that measurement of sCD44v5 might be useful in monitoring RA+ patients with CYA.

Increased sequestration of organic carbon in soil by hydrophobic protection

The impact of soil organic carbon dynamics on the global carbon cycle is still largely uncertain despite studies of agricultural activities and control emissions of greenhouse gases to the earth's atmosphere. Improved knowledge of organic matter dynamics should lead to reduction in CO(2 )emissions. We used stable carbon isotope analysis to detect small changes in organic carbon storage and turnover upon soil treatments with a (13)C-labeled aliphatic alcohol previously partitioned into soluble humic substances of varying hydrophobicity. We found that labeled organic carbon is increasingly protected from mineralization with increased hydrophobic character of humic matter. The stabilization of organic carbon by hydrophobic protection significantly reduced decomposition during incubation time in soil. Hydrophobic protection can become an useful tool to limit decomposition of fresh organic matter in soil and thus reduce CO(2) emission from agricultural soils on a global scale.http://link. springer.de/link/service/journals/00114/bibs/9086010/90860496. htm

Effects of elevated ozone concentration on the degradation of dichlorprop in soil.

An aerobic degradation study was conducted to estimate possible effects of elevated ozone concentration in air on the behaviour of dichlorprop. An average ozone concentration of 80 nL L-1 was chosen, which often occurs close to congested areas during late spring and summer. A control soil and an ozone exposed soil were kept under same conditions such as temperature, air flow and soil humidity. The use of 14C-labelled dichlorprop allowed to examine the fate of dichlorprop and follow the degradation products in soil. Exhaustive extraction of both soils yielded several fractions containing dichlorprop residues. Half lives of dichlorprop of both treatments were 5 days. After 32 days most of the residues in soil remained in the non extractable fraction. The elevated ozone concentration showed no significant effects on the degradation behaviour of dichlorprop and its metabolites but significant differences were found for the behaviour of the nonextractable residues and of the release of carbon dioxide, which were higher for control soil in comparison to the ozone variant. These findings suggest that even moderately elevated ozone concentration in air can effect mineralisation and fixation processes of dichlorprop.

A simple cleanup method for the isolation of nitrate from natural water samples for O isotope analysis.

The analysis of O-isotopic composition of nitrate has many potential applications in studies of environmental processes. O-isotope nitrate analysis requires samples free of other oxygen-containing compounds. More than 100% of non-NO3- oxygen relative to NO3- oxygen can still be found in forest soil water samples after cleanup if improper cleanup strategies, e.g., adsorption onto activated carbon, are used. Such non-NO3- oxygen compounds will bias O-isotopic data. Therefore, an efficient cleanup method was developed to isolate nitrate from natural water samples. In a multistep cleanup procedure using adsorption onto water-insoluble poly(vinylpyrrolidone), removal of almost all other oxygen-containing compounds, such as fulvic acids, and isolation of nitrate was achieved. The method supplied samples free of non-NO3- oxygen which can be directly combusted to CO2 for subsequent O-isotope analysis.

Serum levels of soluble CD44 variant isoforms are elevated in rheumatoid arthritis.

Serum levels of soluble CD44 variant proteins including sequences encoded by exon v5 and exon v6 (sCD44v5, sCD44v6) were determined in patients with inflammatory rheumatic diseases: 56 with rheumatoid arthritis (RA+) and 31 with miscellaneous inflammatory rheumatic diseases (MIRD). There were very significantly higher serum levels of sCD44v5 and sCD44v6 in patients with RA+ than in those with MIRD (RA+ to MIRD: sCD44v5: 81 +/- 54 ng/ml to 33 +/- 13 ng/ml; sCD44v6: 237 +/- 124 ng/ml to 166 +/- 53 ng/ml; both P << 0.001). In RA+ elevated serum levels of sCD44v5 were correlated with the inflammatory activity of disease. In 17 patients with RA+ three or four follow-up measurements of sCD44v5 were performed within 6 months. The development of sCD44v5 serum levels reflected the clinical course of disease in the patients investigated.

Increased serum levels of soluble CD44-isoform v5 in rheumatic diseases are restricted to seropositive rheumatoid arthritis.

Serum levels of sCD44v5 were measured in 134 patients with definite inflammatory rheumatic diseases (IRD) using a sandwich type ELISA. 94 patients suffered from erosive IgM-rheumatoid factor positive rheumatoid arthritis (RA+), 20 with undifferentiated seronegative polyarthritis, 12 with osteoarthropathia psoriatica and psoriasis vulgaris, 3 with systemic lupus erythematosus, 3 with scleroderma and 2 with reactive arthritis. Elevated serum levels (> 58 ng/ml to 221 ng/ml; median: 93 ng/ml) were only detected in 54/94 (57%) patients with RA+, but not in other IRD. They correlated with advanced stages of disease (Steinbrocker stages III + IV; p < 0.05), elevated CRP-levels (p < 0.01) and higher measurements of IgM rheumatoid factor.

[Clinical significance of autologous transplantation with hematopoietic stem cells in leukemia and solid tumors]. / Die klinische Bedeutung der autologen Transplantation mit hämatopoetischen Stammzellen bei Leukämien und soliden Tumoren.

Autologous Transplantation of hematopoietic tissue with frozen hematopoietic stem cells is increasingly used for leukemias and lymphomas, but also for some solid tumors. In the past, autotransplants have been performed with bone marrow as the source of hematopoietic stem cells. Circulating, blood derived hematopoietic stem cells, however, allow safe engraftment of all cell lines after supralethal chemo-radiotherapy. This survey describes the role of autologous stem cell transplantation in disorders that are currently in the center of clinical and scientific interest. This estimation is based on the proportion of protocols dealing with, and centering on, autologous stem cell transplantation in the context of treatment for leukemias and solid tumors ("Oncodisc", "PDQ").

[Epidemiology of attempted suicide by drug overdose in Vienna]. / Beitrag zur Epidemiologie medikamentöser Suizidversuche in Wien.

We studied 361 Viennese patients (m = 111, f = 250) admitted to our observation ward after attempted suicide by drug ingestion over a two-year period. Age and sex distribution, social status, time of attempt, motives, and drugs used were documented, as well as the therapeutic management of the cases. The patients were divided into 4 age groups: 57 juveniles, 179 young adults, 80 middle-aged adults and 45 geriatric patients. These groups were compared with each other in manifold respects. Our analysis showed that the geriatric patients who had attempted suicide by drug ingestion differed from those of other groups insofar as there was a lack of exogenous influence with regard to motives and methods. They may represent a separate entity more akin to suicide than to attempted suicide. Vienna data on attempted suicide by drug ingestion did not differ markedly from international data.