Importance of species‐specific antigens in the serodiagnosis of Chlamydia trachomatis reactive arthritis

Objectives. To determine the most sensitive and specific method of anti‐Chlamydia antibody measurement for the serodiagnosis of Chlamydia trachomatis reactive arthritis. Methods. Immunoblotting, enzyme‐linked immunosorbent assays using six synthetic peptides or recombinant antigens and a microimmunofluorescence test were used to determine the presence of IgG, IgM and IgA in serum samples from 17 patients with C. trachomatis reactive arthritis. Twenty patients with other inflammatory arthropathies without evidence of urogenital C. trachomatis infection were used as controls. Results. The best association of sensitivity (76%) and specificity (85%) was obtained when IgG and/or IgA reactivity to two species‐specific antigens was determined. These antigens were synthetic peptides, derived from species‐specific epitopes in the variable domain IV of the major outer membrane protein (MOMP) (Labsystems, Finland) and recombinant polypeptide encoded by open reading frame 3 of the plasmid (pgp3). Conclusions. IgG and/or IgA anti‐MOMP‐derived peptides and anti‐pgp3 could be useful for the diagnosis of probable C. trachomatis reactive arthriti

Many--Particle Correlations in Relativistic Nuclear Collisions

Many--particle correlations due to Bose-Einstein interference are studied in ultrarelativistic heavy--ion collisions. We calculate the higher order correlation functions from the 2--particle correlation function by assuming that the source is emitting particles incoherently. In particular parametrizations of and relations between longitudinal, sidewards, outwards and invariant radii and corresponding momenta are discussed. The results are especially useful in low statistics measurements of higher order correlation functions. We evaluate the three--pion correlation function recently measured by NA44 and predict the 2--pion--2--kaon correlation function. Finally, many particle Coulomb corrections are discussed.Comment: 5 corrected misprints, 14 pages, revtex, epsfig, 6 figures included, manuscript also available at http://www.nbi.dk/~vischer/publications.htm

Visualization of SpoVAEa Protein Dynamics in Dormant Spores of <i>Bacillus cereus</i> and Dynamic Changes in Their Germinosomes and SpoVAEa during Germination

Bacillus cereus spores, like most Bacillus spores, can survive for years and germinate when their surroundings become suitable, and germination proteins play an important role in the initiation of germination. Because germinated spores lose the extreme resistance of dormant spores, information on the function of germination proteins could be useful in developing new strategies to control B. cereus spores. Prior work has shown that (i) the channel protein SpoVAEa exhibits high-frequency movement in the outer leaflet of the inner membrane (IM) in dormant B. subtilis spores and (ii) the formation of the foci termed germinosomes between two germination proteins, the germinant receptor GerR and the scaffold protein GerD, in developing B. cereus spores is slower than foci formation by GerR and GerD individually. However, the movement dynamics of SpoVAEa in B. cereus spores, and the behavior of the germinosome upon B. cereus spore germination, are not known. In this study, we found that SpoVAEa fluorescent foci in dormant B. cereus spores move on the IM, but slower than in B. subtilis spores, and they likely co-localize transiently with GerD-mScarlet-I in the germinosome. Our results further indicate that (i) the expression of GerR-SGFP2 and SpoVAEa-SGFP2 with GerD-mScarlet-I from a plasmid leads to more heterogeneity and lower efficiency of spore germination in B. cereus, and (ii) germinosome foci observed by Fluorescence resonance energy transfer (FRET) between GerR-SGFP2 and GerD-mScarlet-I can be lost soon after the spore-phase transition. However, this is not always the case, as some GerR-SGFP2 and GerD-mScarlet-I foci continued to exist, co-localize, and even show a weak FRET signal. These data highlight the heterogeneous behavior of spore germination protein complexes and indicate that some complexes may persist beyond the initiation of germination. IMPORTANCE Bacillus cereus is commonly present in soil and infects humans via contaminated food. In this study, we used B. cereus spores to investigate the movement of the spore-specific inner membrane (IM) channel protein SpoVAEa, the interaction between SpoVAEa and the germinosome scaffold protein GerD, and the dynamics of germinosomes with GerR and GerD in spore germination. Our results expand upon observations of interactions between specific B. cereus spore germination proteins, in particular the GerR germinant receptor A, B, and C subunits and GerD, as well as those between SpoVAEa and GerD. The approaches used in this work could also be used to examine the interactions between GerD and SpoVAEa and other germination proteins in spores of other Bacillus species

Organization and dynamics of the SpoVAEa protein and its surrounding inner membrane lipids, upon germination of <i>Bacillus</i> <i>subtilis</i> spores

The SpoVA proteins make up a channel in the inner membrane (IM) of Bacillus subtilis spores. This channel responds to signals from activated germinant receptors (GRs), and allows release of Ca(2+)-DPA from the spore core during germination. In the current work, we studied the location and dynamics of SpoVAEa in dormant spores. Notably, the SpoVAEa-SGFP2 proteins were present in a single spot in spores, similar to the IM complex formed by all GRs termed the germinosome. However, while the GRs’ spot remains in one location, the SpoVAEa-SGFP2 spot in the IM moved randomly with high frequency. It seems possible that this movement may be a means of communicating germination signals from the germinosome to the IM SpoVA channel, thus stimulating CaDPA release in germination. The dynamics of the SpoVAEa-SGFP2 and its surrounding IM region as stained by fluorescent dyes were also tracked during spore germination, as the dormant spore IM appeared to have an immobile germination related functional microdomain. This microdomain disappeared around the time of appearance of a germinated spore, and the loss of fluorescence of the IM with fluorescent dyes, as well as the appearance of peak SpoVAEa-SGFP2 fluorescent intensity occurred in parallel. These observed events were highly related to spores’ rapid phase darkening, which is considered as due to rapid Ca(2+)DPA release. We also tested the response of SpoVAEa and the IM to thermal treatments at 40–80 °C. Heat treatment triggered an increase of green autofluorescence, which is speculated to be due to coat protein denaturation, and 80 °C treatments induce the appearance of phase-grey-like spores. These spores presumably have a similar intracellular physical state as the phase grey spores detected in the germination but lack the functional proteins for further germination events

SYNOVIAL FLUID AND SERUM ANTIBODIES AGAINST CHLAMYDIA IN DIFFERENT FORMS OF ARTHRITIS: INTRA-ARTICULAR IgA PRODUCTION IN CHLAMYDIA SEXUALLY ACQUIRED REACTIVE ARTHRITIS

SUMMARY Since the presence of Chlamydia has been shown in synovial fluid (SF) from some patients with Chlamydia reactive arthritis, we investigated whether anti-Chlamydia antibodies present in the joint are derived from the circulation or are locally produced. We compared titres of IgG, IgM and IgA antibodies against Chlamydia, and against a control antigen (tetanus toxoid), by an enzyme-linked immunosorbent assay (ELISA), in paired samples of serum and SF from Chlamydia trachomatis sexually acquired reactive arthritis (CT-SARA) patients and from patients with other forms of arthritis. The ratio of serum/SF IgA anti-Chlamydia antibodies was significantly decreased in CT-SARA patients. It is concluded that, in our experimental conditions, we found evidence for intra-articular production of IgA anti-Chlamydia antibodie

THE VALUE OF ISOTYPE DETERMINATION OF SERUM ANTIBODIES AGAINST CHLAMYDIA FOR THE DIAGNOSIS OF CHLAMYDIA REACTIVE ARTHRITIS

SUMMARY In clinical rheumatology, the diagnosis of Chlamydia reactive arthritis is difficult because an incomplete form of the disease can closely resemble an undifferentiated seronegative mono/oligoarthritis. We investigated whether measuring specific isotypes of anti-Chlamydia antibodies in serum can improve the diagnosis, by comparing such antibody concentrations in the serum of patients with well-defined disease, i.e. Chlamydia trachomatis sexually acquired reactive arthritis (CT-SARA), with other arthritides. Antibody levels were determined by enzyme-linked immunosorbent assay (ELISA). When considering two different isotypes and their combination, the best sensitivity (63%) was obtained for IgM and/or IgA results with a specificity of 81%. The patients with CT-SARA and SARA had the highest levels of antibodies of all isotypes tested. It is concluded that, in our experimental conditions, only very high values of specific isotypes could indicate a diagnosis of Chlamydia reactive arthriti