Anti-citrullinated fibronectin antibodies in rheumatoid arthritis are associated with human leukocyte antigen-DRB1 shared epitope alleles

Introduction

Fibronectin is one of the most abundant proteins present in the inflamed joint. Here, we characterized the citrullination of fibronectin in the joints of rheumatoid arthritis (RA) patients and studied the prevalence, epitope specificity and human leukocyte antigen (HLA) association of autoantibodies against citrullinated fibronectin in RA.

Methods

Citrullinated residues in fibronectin isolated from RA patient synovial fluid were identified by mass spectrometry. The corresponding citrullinated and non-citrullinated peptides were synthesized and used to analyze the presence of autoantibodies to these peptides in RA sera and sera from other diseases and healthy controls by ELISA. The data were compared with risk factors like shared epitope HLA alleles and smoking, and with clinical features.

Results

Five citrullinated residues were identified in fibronectin from RA synovial fluid. RA sera reacted in a citrulline-dependent manner with two out of four citrullinated fibronectin peptides, one of which contains two adjacent citrulline residues, in contrast to non-RA sera, which were not reactive. The most frequently recognized peptide (FN-Cit_1035,1036, LTVGLTXXGQPRQY, in which × represents citrulline) was primarily targeted by anti-CCP (cyclic citrullinated peptide) 2-positive RA patients. Anti-FN-Cit_1035,1036 autoantibodies were detected in 50% of established anti-CCP2-positive RA patients and in 45% of such patients from a early arthritis clinic. These antibodies appeared to be predominantly of the immunoglobulin G (IgG) isotype and to be associated with HLA shared epitope alleles (odds ratio = 2.11).

Conclusions

Fibronectin in the inflamed synovia of RA patients can be citrullinated at least at five positions. Together with the flanking amino acids, three of these citrullinated residues comprise two epitopes recognized by RA autoantibodies. Anti-citrullinated fibronectin peptide antibodies are associated with HLA shared epitope alleles.     

La autoantigen suppresses IRES-dependent translation of the hepatitis A virus

The human RNA-binding protein La, is an essential trans-acting factor in IRES-dependent translation initiation of poliovirus, the prototypic picornavirus. For hepatitis A virus (HAV), an unusual member of this virus family, the role of host proteins in its inefficient translation and slow replication is unclear. Using small interfering RNA in vivo and purified La in vitro, we demonstrate for the first time that La suppresses HAV IRES-mediated translation and replication. We show that La binds specifically to distinct parts of the HAV IRES and that—unlike poliovirus—HAV proteinase 3C does not cleave La. The La-mediated suppression of HAV translation and stimulation of poliovirus translation implies unexpected mechanistic differences between viral IRES elements.     

The influence of upwelling,coastal currents and water temperature on the distribution of the red tide dinoflagellate, <Emphasis Type="Italic">Noctiluca scintillans</Emphasis>, along the east coast of Australia

Quasi-synoptic surveys along the east coast of Australia between 28 and 34°S show that the heterotrophic dinoflagellate, Noctiluca scintillans, occurs along this entire stretch of the coast. Areas of relatively high abundance of Noctiluca were observed downstream of regions predisposed to current-induced upwellings as a consequence of alongshore topographic variations. High-resolution temporal and spatial sampling of upwelling events showed that Noctiluca was abundant (up to 28 cells l^?1) within mature upwelled waters. A high proportion (>80%) of fed Noctiluca cells (cells with prey in their vacuoles) was observed in the mature upwelled waters indicating that the observed increase in abundance of Noctiluca was associated with increased feeding activity. The absolute abundance of Noctiluca in upwelled waters was, however, found to vary from one upwelling location to another and between seasons. In particular, highest abundances of Noctiluca were recorded south of 31.5°S, where the East Australian Current (EAC) characteristically separates from the coast. The high abundances partly arise from southward advection and retention of the Noctiluca cells, and partly from upwelling inshore of the separated EAC driven by cross-shelf boundary layer fluxes. The temperature of the EAC was also found to influence absolute abundances. Surface water temperatures during our summer cruise were anomalously high due to a strong La Niña phase, and up to 4°C warmer than during our spring cruise. We found that the warmer surface water temperatures were associated with relatively lower average abundances of Noctiluca in the near shore zone.     

广东鹤山人工林群落主要优势植物的热值和灰分含量

从广东鹤山南亚热带丘陵5种人工林群落中采集20种主要植物器官及3种群落凋落叶样品,利用PARR-1281氧弹热量计和马福炉分别测定样品的干质量热值和灰分含量,再计算去灰分热值;按器官、个体和生长型等比较分析植物的热值和灰分特征.结果表明：植物各器官的干质量热值、去灰分热值分别在10.7～22.17和13.89～23.04 kJ·g^-1之间,叶片的干质量热值和去灰分热值明显高于其他器官(P＜0.05).个体加权平均干质量热值和去灰分热值分别在14.24～19.43和16.63～20.99 kJ·g^-1群落各层次植物去灰分热值平均值的高低依次为：乔木(19.55 kJ·g^-1)＞灌木(19.46 kJ·g^-1)＞草本(18.77 kJ·g^-1),其中,乔木层树木的去灰分热值平均值大小为：本地针叶树(19.86 kJ·g^-1)＞本地阔叶树(19.55 kJ·g^-1)＞外来桉树(19.18 kJ·g^-1),灰分含量的排序则相反.马占相思林、针叶林和木荷林群落的凋落叶热值均高于所在林分各层次的热值(P＜0.01),马占相思林和针叶林的凋落叶热值大于所在乔木层叶片的热值,而木荷林凋落叶热值小于乔木层叶片的热值.     

Effects of Rare Earth Elements on the Growth of Arnebia euchroma Cells and the Biosynthesis of Shikonin

Nd³⁺, La³⁺ and Ce³⁺ at proper concentrations had positive effects on the cell growth of Arnebia euchroma and production of shikonin derivatives. A mixture of rare earth elements (MRE, La₂O₃:CeO₂:Pr₆O₁₁: Sm₂O₃ = 255:175:3:1, mol/mol) behaved the most remarkable effects. Two-stage culture was used for the cell proliferation and the biosynthesis of shikonin derivatives. After 20 days culture, 0.05 mM MRE gave the highest cell biomass (24.8 g dry weight l⁻¹), which was 98.0% higher than that without rare earth elements. Similarly, when 0.05 mM MRE was added to the biosynthesis medium, the highest content (8.9% dry weight) and production (571.1 mg l⁻¹) of shikonin derivatives were obtained, which were 89.4% and 165.3% higher than those without rare earth elements, respectively. The increase of the cell biomass and shikonin derivatives may due to increasing the activities of peroxidase and phenylalanine ammonia lyase caused by the addition of the rare earth elements.     

硝酸镧在小鼠肝中的积累及遗传毒理研究

通过在饮水中加入硝酸镧使小鼠摄入稀土,1个月后采用ICP-MS法测试了镧在小鼠肝中的积累,研究了硝酸镧对骨髓细胞微核率的影响,并采用体外试验研究了硝酸镧对小鼠基因组DNA的切割作用,探讨了稀土元素镧的遗传毒理.结果表明,1 000、500、300和50 μg·ml^-1处理组小鼠肝中镧含量分别达1.46、0.558、0.529和0.083 μg·g^-1,与对照组0.028 μg·g^-1的含量相比,各处理组小鼠肝中镧的含量皆有升高且与喂饮硝酸镧溶液的浓度成正比（r＝0.980）.1 000、500和300 μg·ml^-1处理组微核率与对照组之间的t检验结果表明各组微核率显著上升（P<0.05）,且亦与喂饮硝酸镧溶液的浓度成正比（r＝0.853）.体外试验显示,硝酸镧可切断DNA链,说明稀土元素镧可在生物体内积累,其在细胞内可使DNA断裂而导致遗传物质受损.     

Density-dependent effects on hatching success of the olive ridley turtle, Lepidochelys olivacea

Historically, the olive ridley arribada at Playa Nancite, Costa Rica, was one of the largest olive ridley arribadas in the eastern Pacific with 70,000 nesting females in a year. Recently the Nancite arribada drastically declined. We hypothesized that the population decline at Playa Nancite could have been due to low hatching success as a result of the high density of nests on the beach, such that recruitment to the population was insufficient to balance losses. To test this hypothesis, we examined density-dependent effects on hatching success and their underlying mechanisms by experimentally manipulating nest densities in experimental plots on the nesting beach. We set up four nest-density treatments in five experimental blocks. We measured effects of density on hatching success, CO(2) and O(2) concentrations and temperature both within nests and in sand adjacent to nests frequently during incubation. Experimental nest densities affected hatching success with the highest density having the lowest hatching success. Higher nest density led to lower O(2) levels and higher CO(2) levels in the nest with greater changes in the latter part of the incubation. Highest temperatures occurred in high-density areas. Temperatures were lower in sand surrounding the nest than in the nest. Effects of density on temperature, CO(2) and O(2) were confirmed at a naturally high-density nesting beach, Playa La Flor, Nicaragua. Long-term failure in production of hatchlings due to historic high densities may have contributed to the decline of arribadas on Playa Nancite. Thus, density-dependent population control would have operated at the embryonic life stage in this population of olive ridley turtles.     

A diepitopic sequential oligopeptide carrier (SOCn) as mimic of the sm autoantigen: synthesis, conformation and biological assays.

Anti-Sm (Sm: U1-U6 RNA-protein complex) antibodies are usually considered highly specific for systemic lupus erythematosus (SLE), while anti-U1RNP (U1RNP: U1RNA-protein complex) are thought of as diagnostic criteria for the mixed connective tissue disease (MCTD). However, both antibody specificities coexist in SLE and MCTD, in varying percentages. Although the anti-Sm/anti-U1RNP immunological cross-reactivity has been initially attributed to a common motif, PPXY(Z)PP (where X, Y, Z are various amino acids), found in the Sm, U1-A and U1-C autoantigens, it appears that the conformational features of the Sm epitopes also play an important role in the immunoreactivity. The PPGMRPP and PPGIRGP main epitopes of the Sm antigen were coupled in duplicate to the tetrameric Ac-(Lys-Aib-Gly)4-OH, SOC4, carrier to form the [(PPGMRPP)2, (PPGIRGP)2]-SOC4 construct as a mimic of the native Sm. It was found that: (i) the 3(10) helical structure of SOC4 allows the epitopes to adopt an exposed orientation, similar to their free forms, that facilitates their recognition from the anti-Sm antibodies, and (ii) the U1-RNP cross-reactivity is minimized.