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Associations between topographic location and articular cartilage repair in preclinical animal models are unknown. Based on clinical investigations, we hypothesized that lesions in the ovine femoral condyle repair better than in the trochlea. Full-thickness chondral and osteochondral defects were simultaneously established in the weightbearing area of the medial femoral condyle and the lateral trochlear facet in sheep, with chondral defects subjected to subchondral drilling. After 6 months in vivo, cartilage repair and osteoarthritis development was evaluated by macroscopic, histological, immunohistochemical, and biochemical analyses. Macroscopic and histological articular cartilage repair and type-II collagen immunoreactivity were better in the femoral trochlea, regardless of the defect type. Location-independently, osteochondral defects induced more osteoarthritic degeneration of the adjacent cartilage than drilled chondral lesions. DNA and proteoglycan contents of chondral defects were higher in the condyle, reflecting physiological topographical differences. The results indicate that topographic location dictates the structural patterns and biochemical composition of the repair tissue in sheep. These findings suggest that repair of cartilage defects at different anatomical sites of the ovine stifle joint needs to be assessed independently and that the sheep trochlea exhibits cartilage repair patterns reflective of the human medial femoral condyle. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31:1772–1779, 2013

Background: Marrow stimulation techniques such as subchondral drilling are clinically important treatment options for symptomatic small cartilage defects. Little is known about whether they induce deleterious changes in the subchondral bone. Hypothesis: Subchondral drilling induces substantial alterations of the microarchitecture of the subchondral bone that persist for a clinically relevant postoperative period in a preclinical large animal model. Study Design: Controlled laboratory study. Methods: Standardized full-thickness chondral defects in the medial femoral condyles of 19 sheep were treated by subchondral drilling. Six months postoperatively, the formation of cysts and intralesional osteophytes was evaluated. A standardized methodology was developed to segment the ovine subchondral unit into reproducible volumes of interest (VOIs). Indices of bone structure were determined by micro–computed tomography (micro-CT). Results: Analysis of the microarchitecture revealed the absence of zonal stratification in the ovine subarticular spongiosa, permitting an unimpeded and simultaneous analysis of the entire subchondral trabecular network. Subchondral drilling led to the formation of subchondral bone cysts (63%) and intralesional osteophytes (26%). Compared with the adjacent unaffected subchondral bone, drilling induced significant alterations in nearly all parameters for the microarchitecture of the subchondral bone plate and the subarticular spongiosa, most importantly in bone volume, bone surface/volume ratio, trabecular thickness, separation, pattern factor, and bone mineral density (BMD) (all P ≤ .01). Conclusion: The data show that the ovine subchondral bone can be reliably evaluated using micro-CT with standardized VOIs. We report that subchondral drilling deteriorates the microarchitecture both of the subchondral bone plate and subarticular spongiosa and decreases BMD. These results suggest that the entire osteochondral unit is altered after drilling for an extended postoperative period. Clinical Relevance: The subchondral bone remains fragile after subchondral drilling for longer durations than previously expected. Further evaluations of structural subchondral bone parameters of patients undergoing marrow stimulation are warranted.

The extracellular adherence protein (Eap) from Staphylococcus aureus has been suggested as a vaccine candidate and for therapeutic use due to its immunomodulating and antiangiogenic properties; however, little is known about anti-Eap antibodies in humans. We determined anti-Eap antibody titers by enzyme-linked immunosorbent assay and Western blot and measured serum samples from 92 patients with proven S. aureus infections and 93 healthy controls. The functionality of antibodies was assessed by a phagocytosis assay using Eap-coated fluorescent microspheres. Antibodies were detected in all human samples, but not in mice. Patients showed significantly higher titers than controls [immunoglobulin M (IgM), P=0.007; IgG, P

Background— Oxidative stress is causally linked to the progression of heart failure, and mitochondria are critical sources of reactive oxygen species in failing myocardium. We previously observed that in heart failure, elevated cytosolic Na + ([Na + ] i ) reduces mitochondrial Ca 2+ ([Ca 2+ ] m ) by accelerating Ca 2+ efflux via the mitochondrial Na + /Ca 2+ exchanger. Because the regeneration of antioxidative enzymes requires NADPH, which is indirectly regenerated by the Krebs cycle, and Krebs cycle dehydrogenases are activated by [Ca 2+ ] m , we speculated that in failing myocytes, elevated [Na + ] i promotes oxidative stress. Methods and Results— We used a patch-clamp–based approach to simultaneously monitor cytosolic and mitochondrial Ca 2+ and, alternatively, mitochondrial H 2 O 2 together with NAD(P)H in guinea pig cardiac myocytes. Cells were depolarized in a voltage-clamp mode (3 Hz), and a transition of workload was induced by β-adrenergic stimulation. During this transition, NAD(P)H initially oxidized but recovered when [Ca 2+ ] m increased. The transient oxidation of NAD(P)H was closely associated with an increase in mitochondrial H 2 O 2 formation. This reactive oxygen species formation was potentiated when mitochondrial Ca 2+ uptake was blocked (by Ru360) or Ca 2+ efflux was accelerated (by elevation of [Na + ] i ). In failing myocytes, H 2 O 2 formation was increased, which was prevented by reducing mitochondrial Ca 2+ efflux via the mitochondrial Na + /Ca 2+ exchanger. Conclusions— Besides matching energy supply and demand, mitochondrial Ca 2+ uptake critically regulates mitochondrial reactive oxygen species production. In heart failure, elevated [Na + ] i promotes reactive oxygen species formation by reducing mitochondrial Ca 2+ uptake. This novel mechanism, by which defects in ion homeostasis induce oxidative stress, represents a potential drug target to reduce reactive oxygen species production in the failing heart.

Purpose There is increasing evidence that genetic factors regulating the recognition and/or repair of DNA double-strand breaks (DSBs) are responsible for differences in radiosensitivity among patients. Genetically defined DSB repair capacities are supposed to determine patients' individual susceptibility to develop adverse normal tissue reactions after radiotherapy. In a preclinical murine model, we analyzed the impact of different DSB repair capacities on the cumulative DNA damage in normal tissues during the course of fractionated irradiation. Material and Methods Different strains of mice with defined genetic backgrounds (SCID −/− homozygous, ATM −/− homozygous, ATM +/− heterozygous, and ATM +/+ wild-type mice) were subjected to single (2 Gy) or fractionated irradiation (5 × 2 Gy). By enumerating γH2AX foci, the formation and rejoining of DSBs were analyzed in organs representative of both early-responding (small intestine) and late-responding tissues (lung, kidney, and heart). Results In repair-deficient SCID −/− and ATM −/− homozygous mice, large proportions of radiation-induced DSBs remained unrepaired after each fraction, leading to the pronounced accumulation of residual DNA damage after fractionated irradiation, similarly visible in early- and late-responding tissues. The slight DSB repair impairment of ATM +/− heterozygous mice was not detectable after single-dose irradiation but resulted in a significant increase in unrepaired DSBs during the fractionated irradiation scheme. Conclusions Radiation-induced DSBs accumulate similarly in acute- and late-responding tissues during fractionated irradiation, whereas the whole extent of residual DNA damage depends decisively on the underlying genetically defined DSB repair capacity. Moreover, our data indicate that even minor impairments in DSB repair lead to exceeding DNA damage accumulation during fractionated irradiation and thus may have a significant impact on normal tissue responses in clinical radiotherapy.

Background/Aims: Defective p53-mediated apoptosis and cell cycle control have been implicated in the immunopathogenesis of Crohn’s disease (CD). Since common functional variants of p53 (SNP72 G/C) and its key negative regulator mdm2 (SNP309 T/G) have been reported to affect cellular apoptotic and cell cycle arrest capacities, we assessed the effects of these variants on CD susceptibility and their relationship to NOD2/CARD15 as a well-established genetic CD risk factor. Methods: The variants SNP72 G/C and SNP309 T/G were genotyped in 149 European CD patients and 478 healthy controls. Subgroup analysis was performed in relation to NOD2/CARD15 status and to demographic/clinical characteristics. Results: The p53 SNP72 CC genotype tended to be less frequent in CD. This reached statistical significance only in the male cohort (0 vs. 7.3%; p = 0.037). Genotype and allele frequencies of both single-nucleotide polymorphisms (SNPs) were otherwise not significantly different. In the combined genotypic analysis, the genotype p53 SNP72 CC was significantly underrepresented in mdm2 SNP309 TT homozygotes (0 vs. 9.7%; p = 0.034). No association was observed between NOD2/CARD15 and the respective SNPs. Conclusion: We report on a gender-specific protective effect of the low-apoptotic SNP72 CC genotype, and a gender-unrestricted genotypic interaction between SNP309 TT and SNP72 CC, which, for the first time, links sequence variation of the p53/mdm2 network to CD, independent of NOD2/CARD15.

OBJECTIVE. Recently, transient elastography (FibroScan) has been introduced for noninvasive staging of liver fibrosis. Here, we investigated a novel approach for noninvasive assessment of liver fibrosis using sonography-based real-time elastography, which can be performed with conventional ultrasound probes during a routine sonography examination.MATERIALS AND METHODS. Real-time elastography was performed in 79 patients with chronic viral hepatitis and known fibrosis stage and in 20 healthy volunteers. A specially developed program was used for quantification of tissue elasticity. Stepwise logistic regression analysis was performed to define an elasticity score using variables with high reproducibility in a preceding analysis of data from 16 different patients. In addition, aspartate transaminase-to-platelet ratio index (APRI) and routine laboratory values were included in the analysis.RESULTS. The Spearman's correlation coefficient between the elasticity scores obtained using real-time elastography and t...

Aim To assess the diagnostic value of pre-surgery axillary ultrasound for nodal staging in patients with primary breast cancer and to identify clinical/histopathological factors impacting diagnostic performance. Study Design Single-center, retrospective chart analysis. We assessed sensitivity, specificity, and positive and negative predictive value of clinical examination as well as axillary ultrasound vs. clinical examination alone. The histopathological results were the standard of truth. In addition, we analyzed clinical and histopathological factors regarding their potential to impact sensitivity and specificity. Results We enrolled a total of 172 women in the study. Sensitivity of clinical examination plus ultrasound was significantly higher than for clinical examination alone (58% vs. 31.6%). Specificity and positive predictive value were similar while the negative predictive value increased from 63.4% to 73% when additionally applying ultrasound. Sensitivity and specificity of axillary ultrasound were impacted by tumor size ( P = 0.2/0.04), suspicious axillary palpation ( P < 0.01/Conclusion Since pre-surgery axillary nodal staging is currently used to determine disease management, axillary ultrasound is a useful add-on tool in the diagnostic armamentarium for breast cancer patients. Tumor size, suspicious axillary palpation, number of affected lymph nodes and distant metastases increase diagnostic performance of this diagnostic modality.

Associations between topographic location and articular cartilage repair in preclinical animal models are unknown. Based on clinical investigations, we hypothesized that lesions in the ovine femoral condyle repair better than in the trochlea. Full-thickness chondral and osteochondral defects were simultaneously established in the weightbearing area of the medial femoral condyle and the lateral trochlear facet in sheep, with chondral defects subjected to subchondral drilling. After 6 months in vivo, cartilage repair and osteoarthritis development was evaluated by macroscopic, histological, immunohistochemical, and biochemical analyses. Macroscopic and histological articular cartilage repair and type-II collagen immunoreactivity were better in the femoral trochlea, regardless of the defect type. Location-independently, osteochondral defects induced more osteoarthritic degeneration of the adjacent cartilage than drilled chondral lesions. DNA and proteoglycan contents of chondral defects were higher in the condyle, reflecting physiological topographical differences. The results indicate that topographic location dictates the structural patterns and biochemical composition of the repair tissue in sheep. These findings suggest that repair of cartilage defects at different anatomical sites of the ovine stifle joint needs to be assessed independently and that the sheep trochlea exhibits cartilage repair patterns reflective of the human medial femoral condyle.

Introduction DNA methylation of CpG islands within the promoter region of genes is an epigenetic modification with an important role in the development of cancer and it is typically mediated by DNA methyltransferases (DNMTs). In cancer cells, global hypomethylation of the genome as a whole and regional hypermethylation of CpG islands have been reported. Four groups of DNMTs have been identified: DNMT1, DNMT2 (TRDMT1), DNMT3A and DNMT3B. DNMT2 uses the catalytic mechanism of DNMTs, but does in fact methylate RNA. Little is known about the significance of these genes in human breast cancer. In the study presented herein, we analyzed five distinct DNMT single SNPs with regard to potential associations with breast cancer risk. Case description In this study, we genotyped 221 female Caucasian breast cancer patients and 221 female Caucasian healthy controls, and we used five allele-specific real-time polymerase chain reaction (qPCR) assays. We selected one locus within the DNMT1 gene and two loci within the DNMT3A and DNMT3B genes, respectively. Statistics were calculated using the chi-squared and Fisher’s exact tests, and correlated with clinical parameters such as age, diagnosis, histology, TNM stage, hormonal receptor status, human epidermal growth factor receptor 2 (HER2) status, response to treatment and survival. Statistically significant results were obtained for correlations with the DNMT1 gene. Discussion and Evaluation Five genomic loci within the DNMT1, DNMT3A and DNMT3B genes were assessed. Statistical significance (P = 0.030) was identified for DNMT1 SNP (A201G, rs2228612): six women within the control group were GG homozygous (variant), while this mutation was absent in the breast cancer group. Conclusions We conclude that women with the DNMT1 SNP (A201G, rs2228612) GG homozygous genotype (variant) have a lower risk of developing breast cancer compared to heterozygous or wildtype genotypes. To date, alterations within the DNMT1 gene have not been reported to be associated with cancer in the Caucasian population.

Summary BACKGROUND AND STUDY PURPOSE: High resolution imaging modalities and electroencephalographic studies (EEG) are used in the assessment of children with headaches. We evaluated the role of cerebral MRI (cMRI) and EEG in the initial assessment of children with headache as the chief complaint of initial presentation. METHODS: A retrospective chart analysis was performed at a tertiary University Hospital. RESULTS: 209 patients were included in this study [mean age 11.3 years; male 91 (43.5%); female 118 (56.5%)]. The following types of headaches were seen: Unclassified headache: 23.4%; probable migraine 17.2%, migraine without aura 13.4%, complicated migraine 12.4%, migraine with aura 1.0%; tension-type 15.3%, and cluster headaches 0.5%, and secondary headaches 16.7%. In 93 children (44.5%) abnormal physical/neurological findings were noted (multiple entries possible). On cMRI studies the following findings were seen: Infection of sinuses (7.2%), pineal cysts (2.4%), arachnoidial cyst and Chiari malformation (1.9%), unspecified signal enhancement (1.0%), and pituitary enlargement, inflammatory lesion, angioma, cerebral ischaemia, and intra-cerebral cyst (each 0.5%). Electroencephalographic findings included both focal and generalised abnormal slowing (5.3%) and Spike-wave complexes (3.3%). CONCLUSIONS: Despite abnormal findings on neurological/physical examination in a substantial number of children with headaches, the yield of pathological cMRIs was low. The use of EEG recordings was not contributory to the diagnostic and therapeutic approach. More research is needed to better define those patients who are likely to have an intracranial pathology.

Background: Several clinical trials have reported B vitamins to be associated with osteoporosis. Objective: Our objective was to investigate whether low serum B vitamins are associated with altered structural and biomechanical properties of human bone. Design: Femoral heads of 94 men and women who underwent hip arthroplasty were analyzed by using dual-energy X-ray absorptiometry (DXA), biomechanical testing (indentation method), and histomorphometry. In addition, blood was collected to measure serum concentrations of homocysteine, folate, vitamin B-6, vitamin B-12, the bone formation marker osteocalcin, and the bone resorption marker tartrate-resistant acid phosphatase (TRAP). Measurement outcomes were grouped according to subjects with high and low serum concentrations, respectively, of folate, vitamin B-6, and vitamin B-12 (n = 47 for each group). Results: Histomorphometric analysis showed a significantly lower trabecular thickness and trabecular area in subjects with low serum folate concentrations than in those with high serum folate concentrations and a significantly lower trabecular number in subjects with low serum vitamin B-6 concentrations than in those with high serum vitamin B-6 concentrations. In contrast, we found a comparable trabecular structure in subjects with high and low serum vitamin B-12 concentrations. DXA and biomechanical testing did not show significant differences between subjects with high and low serum B vitamin concentrations. Osteocalcin was significantly lowered in subjects with a low serum B vitamin concentration, whereas there was no association between serum B vitamins and TRAP. Conclusion: The results of the present study indicate that low serum folate and vitamin B-6 concentrations, but not low serum vitamin B-12 concentrations, are associated with an altered morphology of human bone. Am J Clin Nutr 2009;90:1440‐5.

To assess the effects of heavy and very heavy smoking on the rate of small for gestational age (SGA) infants, and to assess socio-economic and regional differences in smoking patterns in pregnant women in Germany.The Neonatal and Perinatal database of the federal state of Saarland, Germany was used to perform a population-based analysis of preterm (32 weeks of gestation) and term (36 weeks of gestation) newborns in 2004-2006. The rate of SGA babies dependent on the amount of tobacco exposure among self-identified smokers and non-smokers were assessed, and distinct maternal risk factors for smoking were evaluated. Our data were compared with the German National Perinatal database.14,593 paired data sets (peripartum/perinatal) were included in this study. The overall rate of smoking during pregnancy was 11.8% with a high percentage of pregnant women smoking 11-20 cigarettes/day (heavy smoker; 4.0%), and20 cigarettes/day (very heavy smoker; 0.6%). Self-identified heavy tobacco use significantly increased the risk for SGA infants (p0.01) in women without uteroplacental insufficiency. Risk factors for smoking included ethnicity (German/Caucasian), socio-economic parameters (single vs. non-single households, status of employment) and age. Smoking pattern and the rate of SGA babies in our cohort differed substantially from the national average.Although the overall rate of smoking appears comparable to previously published data, heavy and very heavy smoking was high in our cohort. Heavy smoking was disproportionately associated with SGA. Preventative measures and strategies should take into consideration socio-economic risk factors as well as regional differences, and should be targeted at distinct subgroups that are especially prone to smoking during pregnancy.

Transient elastography has been studied in a multitude of liver diseases for the staging of liver fibrosis with variable results. A meta-analysis was performed to assess the overall performance of transient elastography for the diagnosis of liver fibrosis and to analyze factors influencing the diagnostic accuracy.Literature databases and international conference abstracts were searched. Inclusion criteria were as follows: evaluation of transient elastography, liver biopsy as reference, and assessment of the area under the receiver operating characteristic curve (AUROC). The meta-analysis was performed using the random-effects model for the AUROC, summary receiver operating curve techniques, as well as meta-regression approaches.Fifty studies were included in the analysis. The mean AUROC for the diagnosis of significant fibrosis, severe fibrosis, and cirrhosis were 0.84 (95% confidence interval [CI], 0.82-0.86), 0.89 (95% CI, 0.88-0.91), and 0.94 (95% CI, 0.93-0.95), respectively. For the diagnosis of significant fibrosis a significant reduction of heterogeneity of the AUROC was found when differentiating between the underlying liver diseases (P.001). Other factors influencing the AUROC were the scoring system used and the country in which the study was performed. Age, body mass index, and biopsy quality did not have a significant effect on the AUROC.Transient elastography can be performed with excellent diagnostic accuracy and independent of the underlying liver disease for the diagnosis of cirrhosis. However, for the diagnosis of significant fibrosis, a high variation of the AUROC was found that is dependent on the underlying liver disease.

Background Progranulin autoantibodies (PGRN) have been previously described in sera of patients with different autoimmune diseases such as ANCA associated vasculitis, large vessel vasculitis, Crohn9s disease, and psoriatic arthritis. Basically, progranulin seems to interact with TNF receptors 1 and 2 causing subsequently antiinflammatory effects. However, the presence of autoantibodies against progranulin was proved to result in lower serum progranulin levels, which is hypothesized to entertain proinflammatory environment due to lack of inhibited TNF alpha activity. Here we conducted a study evaluation the presence of PGRN in patients with rheumatoid arthritis, which is an autoimmune disorder with proven TNF alpha mediated joint inflammation. Objectives Prevalence of serum progranulin autoantibodies in RA with stratification into DAS28 subgroups with prognostic relevance. Methods From Nov 2011 till Dec 2013 sera of 307 RA patients (Saarland, Berlin) were collected and analyized for PGRN with ELISA (as previously published [1]) using a retrospective, blinded, non-randomized, two-center study design. All patients were on treatment with conventional DMARDs (n=193) or TNF inhibitors (n=114) at least for three months. All patients fullfill the ACR criteria from 1987 and the revised diagnostic EULAR/ACR criteria from 2011. RA patients characteristics concerning age, gender, treatment history, radiological joint imaging, and RF-/ACPA-serum status were availiable. DAS28 and HAQ representing RA activity were documented at study inclusion visit completed by the previous DAS28 scores as written in the patient9s chart under the same present antirheumatic treatment as availiable (retrospective analysis of data from three months till two years in the past). Results The overall prevalence of PGRN-Abs in RA patients was 26.6%. The RA patients were further stratifyied according to the lowest DAS 28 score during the observation period under the present treatment into patients with scores higher versus lower than 3.0 representing “good treatment response” or not, respectively. Table 1 demonstrates that the presence of PGRN-Abs is independent risk factor for failing “good treatment response” beyond erosive diesease, RF- and ACPA-positivity, HAQ score, and Larsen score in this setting. Conclusions PGRN-Abs in sera of RA patients seems to be predictive for poor response of conventional DMARDs and biological TNF-alpha blocking agents. However, further prospective studies with larger cohorts of RA patients are required to confirm the results. References Thurner L & Assmann G, Arth Res Ther, 2013 Disclosure of Interest None declared

Background Antibodies against citrullinated proteins (ACPA) have been recognised as the most specific serum marker for rheumatoid arthritis. However, serum autoantibodies such as anti-nuclear antibodies have also been detected in the sera of different lymphatic malignancies without accompanying rheumatologic disease. Therefore, we conducted a study to evaluate the prevalence of ACPA in diffuse large B-cell non-Hodgkin lymphoma (DLBCL). Methods Sera of 395 DLBCL patients and 258 age-matched healthy controls were investigated to evaluate the prevalence of ACPA and RF. ACPA-positive data were stratified into subgroups of RF positivity and established prognostic parameters for DLBCL, including overall survival. In addition, the ACPA serum concentrations levels were compared to an ACPA-positive RA cohort (n = 175). The statistics were performed with χ2 test and Mann- Whitney-U test; Kaplan-Meyer curves (log rank test) were used to analyse the overall survival. P-value

Introduction Psoriatic arthritis (PsA) is a distinctive inflammatory arthritis which may typically develop in a subgroup of individuals suffering from psoriasis. We recently described progranulin autoantibodies (PGRN-Abs) in the sera of patients with different autoimmune diseases including seronegative polyarthritis. In the present study we investigated the occurrence of PGRN-Abs in PsA. Methods PGRN-Abs were determined in 260 patients with PsA, 100 patients with psoriasis without arthritic manifestations (PsC) and 97 healthy controls using a recently described ELISA. PGRN plasma levels were determined from subgroups by a commercially available ELISA-kit. Possible functional effects of PGRN-antibodies were analysed in vitro by tumour necrosis factor (TNF)-α mediated cytotoxicity assays using WEHI-S and HT1080 cells. Results PGRN-Abs were detected with relevant titres in 50/260 (19.23%) patients with PsA, but in 0/100 patients with psoriasis without arthritic manifestations (P = 0.0001). All PGRN-Abs belonged to immunoglobulin G (IgG). PGRN-Abs were significantly more frequent in PsA patients with enthesitis or dactylitis. PGRN-Abs were also more frequent in PsA patients receiving treatment with TNF-α-blockers than in patients treated without TNF-α-blockers (20.8% versus 17.4%; P = 0.016). PGRN plasma levels were significantly lower in PGRN-Ab-positive patients with PsA than in healthy controls and patients with psoriasis without arthritic manifestations (P < 0.001), indicating a neutralizing effect of PGRN-Abs. Moreover cytotoxicity assays comparing PGRN-antibody positive with negative sera from matched patients with PsA, clearly showed a proinflammatory effect of PGRN antibodies. Conclusion Neutralizing PGRN-Abs occur with relevant titres in a subgroup of patients with PsA, but not in patients without arthritic manifestations (PsC). PGRN-Ab-positive patients had more frequent enthesitis or dactylitis. TNF-α-induced cytotoxicity assays demonstrated that the protective effects of progranulin were inhibited by serum containing PGRN-Abs. This suggests that PGRN-Ab might not only be useful as a diagnostic and prognostic marker, but may provide a proinflammatory environment in a subgroup of patients with PsA.

Background Elevation of free light chains (FLC) represents B cell activation and has been detected in the autoantibodies positive rheumatologic disorders such as rheumatoid arthritis (RA) and Sjogren syndrome. Psoriatic arthritis (PsA) is typically not accompanied by autoantibodies, however, B cells have been found in PsA synovium (Ref1). Objectives We have conducted a case control study to evaluate the frequency of elevated serum FLC free kappa and free lamda levels in 210 PsA patients to detect polyclonal or monoclonal aberration. Methods Serum FLC levels were measured using the Freelite immunoassay (The Binding Site, Germany) to detect free kappa (normal range 3.3-19.4 mg/l) and lamda (5.7-26.3). Increased free kappa and free lamda levels were defined as polyclonal, a kappa/lamda ratio 1.65 was defined as monoclonal aberration. As controls served healthy controls (n=50), RA patients (n=75) and B-NHLymphoma (n=288). A subgroup analysis was performed after stratification of PsA patients into age, disease duration, development of joint erosions, biological treatment, elevated TNF alpha serum levels. Results PsA patients showed significantly less frequently polyclonal respectively monoclonal increase of FLC (7.1 and 3.8%) compared to RA (10.7 and 10.7%, **p=0.001) and B-NHLymphoma (11.5 and 10.1%, ***p=0.001), but more than in healthy controls (*p=0.001, figure 1). PsA patients with increased TNF alpha serum levels and patients with erosive joint disease had significantly more frequently polyclonal aberrations. Image/graph Conclusions Subgroups of PsA patients showed polyclonal aberrations measured by serum FLC indicating B cell activation in the course of disease. References Cantete JD, et al. Ectopic lymphoid neogenesis in psoriatic arthritis. ARD 2007;66,720-26 Disclosure of Interest G. Assmann Grant/research support from: THE BINDING SITE, Schwetzingen, Germany, put the FREELITE immunoassay at costfree disposal for G Assmann, S. Zinke: None Declared, J. T. Ney: None Declared, M. F. Ong: None Declared, F. Neumann: None Declared, S. Nold: None Declared, M. Pfreundschuh: None Declared

Background The complex processes of citrullination presents an ubiquitous pathophysiological procedure during inflammation. However, anti-citrullinated protein antibodies (ACPA) are highly specific for the rheumatoid arthritis (RA). In order to a variety of several autoimmune phenomenons during the course of lymphoma diseases the occurrence of autoantibodies like rheumatoid factors (RF) and antinuclear autoantibodies (ANA) has been reported. Objectives Detecting the prevalence of anti-citrullinated protein antibodies (ACPA) in patients with DLCBL and its impact on prognosis including overall survival. Methods We conducted a case control study analyzing 395 sera of patients DLCBL and of 249 healthy controls (HC) to compare the frequency of ACPA-positivity using the χ2-tests for 2×2 tables (with fisher correction as required). In addition, DLCBL patients were stratified into gender, age, stage of lymphoma disease, ESR, LDH elevation, ECOG performance status, and overall survival (analyzed by Mann-Whitney-test and Kaplan-Meyer-curve, respectively). Results The A-CCP have been detected in the sera of 14 DLCBL patients (3.5%) more frequently than in the sera of HC (n=2, 0.8%, p=0.030). The subgroup analysis of female patients in both cohorts showed 4.4% ACPA positive DCBL patients and no HC (p=0.018). Data of DLCBL patients stratified into gender, age $ $60y, stage of disease, performance status, ESR, LDH elevation, and overall survival (OS) show no differences between ACPA positive and negative patients (table). However, there was a trend towards better 6-year OS for ACPA positive cohort (66.79% versus 60.26%). Conclusions The patients with aggressive B-Non-Hodgkin lymphoma (DLCBL) showed more frequently anti-citrullinated protein antibodies (ACPA) compared to healthy controls. However, the ACPA could not been proved as a prognostic marker for DLCBL. Disclosure of Interest None Declared

Abstract 2911 The bone disease in myeloma is the result of the complex interactions between myeloma cells and the bone marrow osteoclasts plus other accessory cells and microenvironmental components. The receptor activator of NfkappaB (RANK) and osteoprotegerin (OPG) cascade system have been reported to be an essential pathway in the osteoclastogenesis. Genetic variations in the genes coding for RANK, RANK ligand (RANKL), and OPG are supposed to play a role in the susceptibility of myeloma. In the present case control study genomic DNA was obtained in 217 myeloma patients including smoldering myeloma (age: mean 62, 33–88) and 516 healthy controls (healthy blood donors, HC; age mean: 44, 18–65). We studied six single-nucleotide-polymorphisms (SNPs) in the genes of RANK (2 SNPs, rs1805034, rs35211496), OPG (2 SNPs, rs3102735, rs2073618), and of RANKL (2 SNPs, rs9533156, rs2277438) using TaqMan assay guided polymerase chain reaction for the respective SNPs. The genotype and allelic frequencies comparing myeloma patients with HC were analyzed with χ2 test for 2×3 and 2×2 tables, respectively. The genotype distributions of the SNP rs2277438 in the RANKL gene and SNP rs3102735 in the OPG gene differed highly significantly (p=0.0001 and p=0.005, respectively) between myeloma patients and HC (table 1). No significantly increased risk was detected in the other SNP analyzed in the study. A subgroup analysis of the myeloma patients stratified into beta2 microglobulin levels lower or higher than 3.5 mg/l showed lower values to be significantly associated with the minor allele of the SNP rs3102735 in the OPG gene (major vs. minor allele in myeloma vs. HC: 77/23% vs. 89/11%; OR 2.49; CI 1.38–4.49). Further stratification into gender, serum calcium levels, hemoglobin values at diagnosis showed no differences.Table 1Myeloma patientsHealthy controls (HC)OR (CI)pRANKL rs2277438Allelen = 434 (%)n = 1024 (%)1324 (75)860 (84)1.7790.00012110 (25)164 (16)(1.373–2.341)Genotypen = 217 (%)n = 512 (%)11132 (61)360 (70)0.00011260 (28)140 (27)2225 (12)12 (2)OPG rs 3102735Allelen = 420 (%)n = 1028 (%)167 (16)109 (11)1.6000.0052353 (84)919 (89)(1.153–2.222)Genotypen = 210 (%)n = 514 (%)119 (4)5 (1)0.0051249 (23)99 (19)22152 (72)410 (80)RANK = receptor activator of NfkappaB, RANKL = receptor activator of NfkappaB ligand, OPG = osteoprotegerin, OR = odds ratio, CI = confidence intervals 95% To the best of our knowledge, this is the first study reporting a highly significant association of the SNP rs2277438 of the RANKL gene and the SNP rs3102735 in the OPG gene, respectively, with the susceptibility to develop myeloma in the Caucasian population. Both SNPs are associated with an increased risk for myeloma. However, myeloma patients carrying the minor allele of OPG SNP rs3102735 developed lower levels of the proliferation marker beta2 microglobulin. The impact of the OPG SNP rs3102735 (location: promoter region, chromosome 8q24) or of the SNP rs2277438 located in intron 1 (chromosome 13q14.11), a potentially regulatory region of the RANKL gene, on the regulation of RANKL expression is unclear. Disclosures: No relevant conflicts of interest to declare.

SummaryObjectiveUpward migration of the subchondral bone plate is associated with osteochondral repair. The aim of this study was to quantitatively monitor the sequence of subchondral bone plate advancement in a lapine model of spontaneous osteochondral repair over a 1-year period and to correlate these findings with articular cartilage repair.DesignStandardized cylindrical osteochondral defects were created in the rabbit trochlear groove. Subchondral bone reconstitution patterns were identified at five time points. Migration of the subchondral bone plate and areas occupied by osseous repair tissue were determined by histomorphometrical analysis. Tidemark formation and overall cartilage repair were correlated with the histomorphometrical parameters of the subchondral bone.ResultsThe subchondral bone reconstitution pattern was cylindrical at 3 weeks, infundibuliform at 6 weeks, plane at 4 and 6 months, and hypertrophic after 1 year. At this late time point, the osteochondral junction advanced 0.19 [95% confidence intervals (CI) 0.10–0.30] mm above its original level. Overall articular cartilage repair was significantly improved by 4 and 6 months but degraded after 1 year. Subchondral bone plate migration correlated with tidemark formation (r = 0.47; P 0.44).ConclusionsThe subchondral bone plate is reconstituted in a distinct chronological order. The lack of correlation suggests that articular cartilage repair and subchondral bone reconstitution proceed at a different pace and that the advancement of the subchondral bone plate is not responsible for the diminished articular cartilage repair in this model.