Analysis of selected pro- and anti-inflammatory cytokines in patients with multiple injuries in the early period after trauma.

INTRODUCTION: Severe trauma causes damage to the protective barriers of the organism, and thus activates immunological reaction. Among substances secreted during this process pro-inflammatory cytokines are of high importance. THE AIM OF THE STUDY: Severe trauma causing multiple injuries is more likely to lead to particularly intensive inflammatory reaction, which can sometimes lead to serious complications, even life-threatening. The aim of the study is to determine those parameters which may serve as predictors of infectious complications and to enable estimation of the patient's immunological status before the decision to introduce elective procedures. MATERIAL AND METHODS: The study population included patients with multiple trauma treated in the Department of Trauma Surgery of the Medical University of Gdansk. The severity of injuries was evaluated with commonly used numerical scales (Revised Trauma Score - RTS, Injury Severity Score - ISS, Glasgow Coma Scale - GCS). Blood samples were collected on the first, second, and fifth day after injury. Evaluated parameters: C-reactive protein (CRP), the level of cytokines: IL-8, IL-1ß, IL-6, TNF, IL-12p70, and IL-10. Control population: individuals without injury. RESULTS: Evaluation of IL-6, IL-8, and CRP levels in patients with multiple trauma in the early period after injury (2-3 days) could be considered as a predictor of delayed infection (5-10 days). CRP level, being cheap and commonly accessible, can be used in clinical practice enabling identification of patients at higher risk of infectious complications and introduction of appropriate treatment and prevention. The analysis of the mentioned parameters may contribute to choosing an appropriate management strategy, including "timing" depending on the patient's biological status.

Associations between single-nucleotide polymorphisms of RFC-1, GGH, MTHFR , TYMS, and TCII genes and the efficacy and toxicity of methotrexate treatment in patients with rheumatoid arthritis.

INTRODUCTION: The differences in drug efficacy and adverse reactions may be caused by genetic variations in drug metabolism between individuals. OBJECTIVES: The aim of the study was to evaluate the effect of gene polymorphisms on the efficacy of therapy and side effects in patients with rheumatoid arthrit s (RA) treated with methotrexate (MTX). PATIENTS AND METHODS: A total of 273 Caucasian patients with RA were treated with MTX for at least 6 months or stopped MTX because of adverse effects. Seven polymorphisms (RFC-1 c.80G>A, GGH c.-401C>T, MTHFR c.1298A>C and c.677C>T, TYMS 2R/3R, TYMS 6-bp deletion, and TCII c.593T>C) were examined for their effects on MTX efficacy and toxicity. Genomic DNA was obtained from peripheral blood leukocytes. RESULTS: Of all patients, 53% reported some adverse effects during at least 1 visit, which led to MTX withdrawal in 17% of the patients. Adverse effects were more frequent in patients with the MTHFR 677T allele than in those with the 677CC genotype (odds ratio [OR], 1.97; P = 0.01) and in those with the GGH 401CC genotype than in those with the GGH 401CT and TT genotypes (OR, 3.8; P = 0.05). Furthermore, the MTHFR 677T allele was associated with increased activity of aminotransferases (OR, 3.4; P = 0.02). MTX-related hepatotoxicity and alopecia were more common in patients with the RFC-1 80AA genotype (OR, 3.5, P = 0.01; OR, 2.4, P = 0.04; respectively). A more rapid positive response to MTX therapy was demonstrated in MTHFR 677CC homozygotes (OR, 3.4; P = 0.001). There were no other associations between single -nucleotide polymorphisms and the efficacy of MTX treatment. CONCLUSIONS: The MTHFR 677CC and GGH 401TT and CT genotypes were associated with a reduction in the number of MTX-related adverse events. Future allele and genotype analyses may help identify the subsets of RA patients with an increased risk of adverse effects.

Isolation of xanthone and benzophenone derivatives from Cyclopia genistoides (L.) Vent. (honeybush) and their pro-apoptotic activity on synoviocytes from patients with rheumatoid arthritis.

A fast and efficient method for the isolation of the C-glucosidated xanthones mangiferin and isomangiferin from the South-African plant Cyclopia genistoides was developed for the first time. The procedure involved extraction, liquid-liquid partitioning with ethyl acetate and subsequent precipitation of mangiferin and isomangiferin from methanol and acetonitrile-water fractions, respectively. Additionally, two benzophenone derivatives: 3-C-ß-glucosides of maclurin and iriflophenone, were isolated from C. genistoides extracts using semi-preparative HPLC. Apart from the above, the isolation procedure also yielded hesperidin and small amounts of luteolin. The structures of the compounds were determined by 1D and 2D NMR experiments and/or LC-DAD-ESI-MS. The selected Cyclopia constituents were screened for pro-apoptotic activity on TNF-α-stimulated synovial cells isolated from rheumatoid arthritis patients. The strongest effect, measured as percent of apoptotic cells, was recorded for isomangiferin (75%), followed by iriflophenone 3-C-ß-glucoside (71%), hesperidin (67%) and mangiferin (65%). The results are encouraging for further studies on the use of the above compounds in the treatment of rheumatoid arthritis.