Analysis of host cell binding specificity mediated by the tp0136 adhesin of the syphilis agent treponema pallidum subsp. Pallidum

Background Syphilis affects approximately 11 million people each year globally, and is the third most prevalent sexually transmitted bacterial infection in the United States. Inability to independently culture and genetically manipulate Treponema pallidum subsp. pallidum, the causative agent of this disease, has hindered our understanding of the molecular mechanisms of syphilis pathogenesis. Here, we used the non-infectious and poorly adherent B314 strain of the Lyme disease-causing spirochete, Borrelia burgdorferi, to express two variants of a known fibronectin-binding adhesin, Tp0136, from T. pallidum SS14 and Nichols strains. Using this surrogate system, we investigated the ability of Tp0136 in facilitating differential binding to mammalian cell lines offering insight into the possible role of this virulence factor in colonization of specific tissues by T. pallidum during infection. Principal findings Expression of Tp0136 could be detected on the surface of B. burgdorferi by indirect immu-nofluorescence assay using sera from a secondary syphilis patient that does not react with intact B314 spirochetes transformed with the empty vector. Increase in Tp0136-mediated adherence of B314 strain to human epithelial HEK293 cells was observed with comparable levels of binding exhibited by both Tp0136 alleles. Adherence of Tp0136-expressing B314 was highest to epithelial HEK293 and C6 glioma cells. Gain in binding of B314 strain expressing Tp0136 to purified fibronectin and poor binding of these spirochetes to the fibro-nectin-deficient cell line (HEp-2) indicated that Tp0136 interaction with this host receptor plays an important role in spirochetal attachment to mammalian cells. Furthermore, preincu-bation of these cell lines with fibronectin-binding peptide from Staphylococcus aureus FnbA-2 protein significantly inhibited binding of B314 expressing Tp0136. Conclusions Our results show that Tp0136 facilitates differential level of binding to cell lines representing various host tissues, which highlights the importance of this protein in colonization of human organs by T. pallidum and resulting syphilis pathogenesis

XMM-Newton observations of the supernova remnant RX J1713.7-3946 and its central source

We present new results from the observations of the supernova remnant (SNR) RX J1713.7–3946 (also G347.3–0.5) performed in five distinct pointings with the EPIC instrument on board the satellite XMM-Newton. RX J1713.7–3946 is a shelltype SNR dominated by synchrotron radiation in the X-rays. Its emission (emission measure and photon index) as well as the absorption along the line-of-sight has been characterized over the entire SNR. The X-ray mapping of the absorbing column density has revealed strong well-constrained variations (0.4 × 1022 cm−2 ≤ NH ≤ 1.1 × 1022 cm−2) and, particularly, a strong absorption in the southwest. Moreover, there are several clues indicating that the shock front of RX J1713.7–3946 is impacting the clouds responsible for the absorption as revealed for instance by the positive correlation between X-ray absorption and X-ray brightness along the western rims. The CO and H observations show that the inferred cumulative absorbing column densities are in excellent agreement with the X-ray findings in different parts of the remnant on condition that the SNR lies at a distance of 1.3 ± 0.4 kpc, probably in the Sagittarius galactic arm, instead of the commonly-accepted value of 6 kpc. An excess in the CO emission is found in the southwest suggesting that the absorption is due to molecular clouds. A search for OH masers in the southwestern region has been unsuccessful, possibly due to the low density of the clouds. The X-ray mapping of the photon index has also revealed strong variations (1.8 ≤ Γ ≤ 2.6). The spectrum is steep in the faint central regions and flat at the presumed shock locations, particularly in the southeast. Nevertheless, the regions where the shock impacts molecular clouds have a steeper spectrum than those where the shock propagates into a low density medium. The search for the thermal emission in RX J1713.7–3946 has been unsuccessful leading to a number density upper limit of 2 × 10−2 cm−3 in the ambient medium. This low density corresponds to a reasonable kinetic energy of the explosion provided that the remnant is less than a few thousand years old. A scenario based on a modified ambient medium due to the effect of a progenitor stellar wind is proposed and leads to an estimate of RX J1713.7–3946’s progenitor mass between 12 and 16 M. The X-ray bright central point source 1WGA J1713.4–3949 detected at the center of SNR RX J1713.7–3946 shows spectral properties very similar to those of the Compact Central Objects found in SNRs and consistent in terms of absorption with that of the central diffuse X-ray emission arising from the SNR. It is highly probable that the point source 1WGA J1713.4–3949 is the compact relic of RX J1713.7–3946’s supernova progenitor.Fil: Cassam Chenaï, G.. Centre D; FranciaFil: Decourchelle, A.. Centre D; FranciaFil: Ballet, J.;. Centre D; FranciaFil: Sauvageot, J. L.. Centre D; FranciaFil: Dubner, Gloria Mabel. Consejo Nacional de Investigaciónes Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Giacani, Elsa Beatriz. Consejo Nacional de Investigaciónes Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; Argentin

A new study towards PSR J1826-1334 and PSR J1826-1256 in the region of HESS J1825-137 and HESS J1826-130

Aims. The goal of this paper is to detect synchrotron emission from the relic electrons of the crushed pulsar wind nebula (PWN) HESS J1825-137 and to investigate the origin of the $\gamma$-ray emission from HESS J1826-130.\\ Methods. The study of HESS J1825-137 was carried out on the basis of new radio observations centred at the position of PSR J1826-1334 performed with the Karl G. Jansky Very Large Array at 1.4 GHz in configurations B and C. To investigate the nature of HESS J1826-130, we reprocessed unpublished archival data obtained with XMM-Newton.\\ Results. The new radio continuum image towards PSR J1826-1334 reveals a bright radio source, with the pulsar located in its centre, which suggests that this feature could be the radio counterpart of the compact component of the PWN detected at high energy. The new 1.4 GHz radio data do not reveal emission with an extension comparable with that observed in $\gamma$-rays for the HESS J1825-137 source. On the other hand, the XMM-Newton study of the region including PSR J1826-1256 reveals an elongated non-thermal X-ray emitting nebula with the pulsar located in the northern border and a tail towards the peak of the very high energy source. The spectrum is characterized by a power law with a photon index going from 1.6 around the pulsar to 2.7 in the borders of the nebula, a behaviour consistent with synchrotron cooling of electrons. From our X-ray analysis we propose that HESS J1826-130 is likely produced by the PWN powered by PSR J1826-1256 via the inverse Compton mechanism.Comment: 9 pages, 5 figures, 4 tables. Accepted for publication in the Astronomy & Astrophysics Journal. The quality of the images has been degrade

The Pulsar Wind Nebula Around PSR B1853+01 in the Supernova Remnant W44

We present radio observations of a region in the vicinity of the young pulsar PSR B1853+01 in the supernova remnant W44. The pulsar is located at the apex of an extended feature with cometary morphology. We argue on the basis of its morphology and its spectral index and polarization properties that this is a synchrotron nebula produced by the spin down energy of the pulsar. The geometry and physical parameters of this pulsar-powered nebula and W44 are used to derive three different measures of the pulsar's transverse velocity. A range of estimates between 315 and 470 km/s are derived, resulting in a typical value of 375 km/s. The observed synchrotron spectrum from radio to X-ray wavelengths is used to put constraints on the energetics of the nebula and to derive the parameters of the pulsar wind.Comment: ApJ Let (in press

Footprint of Positive Selection in Treponema pallidum subsp. pallidum Genome Sequences Suggests Adaptive Microevolution of the Syphilis Pathogen

In the rabbit model of syphilis, infection phenotypes associated with the Nichols and Chicago strains of Treponema pallidum (T. pallidum), though similar, are not identical. Between these strains, significant differences are found in expression of, and antibody responses to some candidate virulence factors, suggesting the existence of functional genetic differences between isolates. The Chicago strain genome was therefore sequenced and compared to the Nichols genome, available since 1998. Initial comparative analysis suggested the presence of 44 single nucleotide polymorphisms (SNPs), 103 small (≤3 nucleotides) indels, and 1 large (1204 bp) insertion in the Chicago genome with respect to the Nichols genome. To confirm the above findings, Sanger sequencing was performed on most loci carrying differences using DNA from Chicago and the Nichols strain used in the original T. pallidum genome project. A majority of the previously identified differences were found to be due to errors in the published Nichols genome, while the accuracy of the Chicago genome was confirmed. However, 20 SNPs were confirmed between the two genomes, and 16 (80.0%) were found in coding regions, with all being of non-synonymous nature, strongly indicating action of positive selection. Sequencing of 16 genomic loci harboring SNPs in 12 additional T. pallidum strains, (SS14, Bal 3, Bal 7, Bal 9, Sea 81-3, Sea 81-8, Sea 86-1, Sea 87-1, Mexico A, UW231B, UW236B, and UW249C), was used to identify “Chicago-“ or “Nichols -specific” differences. All but one of the 16 SNPs were “Nichols-specific”, with Chicago having identical sequences at these positions to almost all of the additional strains examined. These mutations could reflect differential adaptation of the Nichols strain to the rabbit host or pathoadaptive mutations acquired during human infection. Our findings indicate that SNPs among T. pallidum strains emerge under positive selection and, therefore, are likely to be functional in nature

Microlensing by Compact Objects associated to Gas Clouds

We investigate gravitational microlensing of point-like lenses surrounded by diffuse gas clouds. Besides gravitational bending, one must also consider refraction and absorption phenomena. According to the cloud density, the light curves may suffer small to large deviations from Paczynski curves, up to complete eclipses. Moreover, the presence of the cloud endows this type of microlensing events with a high chromaticity and absorption lines recognizable by spectral analysis. It is possible that these objects populate the halo of our galaxy, giving a conspicuous contribution to the fraction of the baryonic dark matter. The required features for the extension and the mass of the cloud to provide appreciable signatures are also met by several astrophysical objects.Comment: 11 pages with 4 figures. Accepted by A&

Transcription of TP0126, Treponema pallidum Putative OmpW Homolog, Is Regulated by the Length of a Homopolymeric Guanosine Repeat

An effective mechanism for introduction of phenotypic diversity within a bacterial population exploits changes in the length of repetitive DNA elements located within gene promoters. This phenomenon, known as phase variation, causes rapid activation or silencing of gene expression and fosters bacterial adaptation to new or changing environments. Phase variation often occurs in surface-exposed proteins, and in Treponema pallidum subsp. pallidum, the syphilis agent, it was reported to affect transcription of three putative outer membrane protein (OMP)-encoding genes. When the T. pallidum subsp. pallidum Nichols strain genome was initially annotated, the TP0126 open reading frame was predicted to include a poly(G) tract and did not appear to have a predicted signal sequence that might suggest the possibility of its being an OMP. Here we show that the initial annotation was incorrect, that this poly(G) is instead located within the TP0126 promoter, and that it varies in length in vivo during experimental syphilis. Additionally, we show that TP0126 transcription is affected by changes in the poly(G) length consistent with regulation by phase variation. In silico analysis of the TP0126 open reading frame based on the experimentally identified transcriptional start site shortens this hypothetical protein by 69 amino acids, reveals a predicted cleavable signal peptide, and suggests structural homology with the OmpW family of porins. Circular dichroism of recombinant TP0126 supports structural homology to OmpW. Together with the evidence that TP0126 is fully conserved among T. pallidum subspecies and strains, these data suggest an important role for TP0126 in T. pallidum biology and syphilis pathogenesis

Whole genome sequence of the Treponema pallidum subsp. endemicum strain Bosnia A: The genome is related to yaws treponemes but contains few loci similar to syphilis treponemes

T. pallidum subsp. endemicum (TEN) is the causative agent of bejel (also known as endemic syphilis). Clinical symptoms of syphilis and bejel are overlapping and the epidemiological context is important for correct diagnosis of both diseases. In contrast to syphilis, caused by T. pallidum subsp. pallidum (TPA), TEN infections are usually spread by direct contact or contaminated utensils rather than by sexual contact. Bejel is most often seen in western Africa and in the Middle East. The strain Bosnia A was isolated in 1950 in Bosnia, southern Europe.The complete genome of the Bosnia A strain was amplified and sequenced using the pooled segment genome sequencing (PSGS) method and a combination of three next-generation sequencing techniques (SOLiD, Roche 454, and Illumina). Using this approach, a total combined average genome coverage of 513× was achieved. The size of the Bosnia A genome was found to be 1,137,653 bp, i.e. 1.6-2.8 kbp shorter than any previously published genomes of uncultivable pathogenic treponemes. Conserved gene synteny was found in the Bosnia A genome compared to other sequenced syphilis and yaws treponemes. The TEN Bosnia A genome was distinct but very similar to the genome of yaws-causing T. pallidum subsp. pertenue (TPE) strains. Interestingly, the TEN Bosnia A genome was found to contain several sequences, which so far, have been uniquely identified only in syphilis treponemes.The genome of TEN Bosnia A contains several sequences thought to be unique to TPA strains; these sequences very likely represent remnants of recombination events during the evolution of TEN treponemes. This finding emphasizes a possible role of repeated horizontal gene transfer between treponemal subspecies in shaping the Bosnia A genome