The Group B Streptococcal Adhesin BspC Interacts with Host Cytokeratin 19 To Promote Colonization of the Female Reproductive Tract

Streptococcus agalactiae, otherwise known as Group B Streptococcus (GBS), is an opportunistic pathogen that vaginally colonizes approximately one third of healthy women. During pregnancy, this can lead to in utero infection, resulting in premature rupture of membranes, chorioamnionitis, and stillbirths. Furthermore, GBS causes serious infection in newborns, including sepsis, pneumonia, and meningitis. Previous studies have indicated that GBS antigen (Ag) I/II family proteins promote interaction with vaginal epithelial cells; thus, we hypothesized that the Ag I/II Group B streptococcal surface protein C (BspC) contributes to GBS colonization of the female reproductive tract (FRT). Here, we show that a ΔbspC mutant has decreased bacterial adherence to vaginal, ecto-, and endocervical cells, as well as decreased auto-aggregation and biofilm-like formation on cell monolayers. Using a murine model of vaginal colonization, we observed that the ΔbspC mutant strain exhibited a significant fitness defect compared to wild-type (WT) GBS and was less able to ascend to the cervix and uterus in vivo, resulting in reduced neutrophil chemokine signaling. Furthermore, we determined that BspC interacts directly with the host intermediate filament protein cytokeratin 19 (K19). Surface localization of K19 was increased during GBS infection, and interaction was mediated by the BspC variable (V) domain. Finally, mice treated with a drug that targets the BspC V-domain exhibited reduced bacterial loads in the vaginal lumen and reproductive tissues. These results demonstrate the importance of BspC in promoting GBS colonization of the FRT and that it may be targeted therapeutically to reduce GBS vaginal persistence and ascending infection

Bovine Staphylococcus aureus superantigens stimulate the entire T 2 cell repertoire of cattle.

Superantigens (SAgs) represent a diverse family of bacterial toxins that induce Vβ-specific T cell proliferation associated with an array of important diseases in humans and animals, including mastitis of dairy cows. However, an understanding of the diversity and distribution of SAg genes among bovine Staphylococcus aureus and their role in the pathogenesis of mastitis is lacking. Population genomic analysis of 195 bovine S. aureus isolates representing 57 unique sequence types revealed that strains encode 2 to 12 distinct SAgs and that the majority of isolates contain 5 or more SAg genes. A genome-scale analysis of bovine reference strain RF122 revealed a complement of 11 predicted SAg genes, which were all expressed in vitro. Detection of specific antibodies in convalescent cows suggests expression of 7 of 11 SAgs during natural S. aureus infection. We determined the Vβ T cell activation profile for all functional SAgs encoded by RF122 revealing evidence for bovine host-specific activity among recently identified RF122-encoded SAgs SElY and SElZ. Remarkably, we discovered that some strains have evolved the capacity to stimulate the entire T-cell repertoire of cattle through an array of diverse SAgs suggesting a key role in bovine immune evasion

Shallow water marine sediment bacterial community shifts along a natural CO2 gradient in the Mediterranean Sea off Vulcano, Italy.

The effects of increasing atmospheric CO(2) on ocean ecosystems are a major environmental concern, as rapid shoaling of the carbonate saturation horizon is exposing vast areas of marine sediments to corrosive waters worldwide. Natural CO(2) gradients off Vulcano, Italy, have revealed profound ecosystem changes along rocky shore habitats as carbonate saturation levels decrease, but no investigations have yet been made of the sedimentary habitat. Here, we sampled the upper 2 cm of volcanic sand in three zones, ambient (median pCO(2) 419 μatm, minimum Ω(arag) 3.77), moderately CO(2)-enriched (median pCO(2) 592 μatm, minimum Ω(arag) 2.96), and highly CO(2)-enriched (median pCO(2) 1611 μatm, minimum Ω(arag) 0.35). We tested the hypothesis that increasing levels of seawater pCO(2) would cause significant shifts in sediment bacterial community composition, as shown recently in epilithic biofilms at the study site. In this study, 454 pyrosequencing of the V1 to V3 region of the 16S rRNA gene revealed a shift in community composition with increasing pCO(2). The relative abundances of most of the dominant genera were unaffected by the pCO(2) gradient, although there were significant differences for some 5 % of the genera present (viz. Georgenia, Lutibacter, Photobacterium, Acinetobacter, and Paenibacillus), and Shannon Diversity was greatest in sediments subject to long-term acidification (>100 years). Overall, this supports the view that globally increased ocean pCO(2) will be associated with changes in sediment bacterial community composition but that most of these organisms are resilient. However, further work is required to assess whether these results apply to other types of coastal sediments and whether the changes in relative abundance of bacterial taxa that we observed can significantly alter the biogeochemical functions of marine sediments

Cu–Ni nanoalloy phase diagram – Prediction and experiment

The Cu-Ni nanoalloy phase diagram respecting the nanoparticle size as an extra variable was calculated by the CALPHAD method. The samples of the Cu-Ni nanoalloys were prepared by the solvothermal synthesis from metal precursors. The samples were characterized by means of dynamic light scattering (DLS), infrared spectroscopy (IR), inductively coupled plasma optical emission spectroscopy (ICP/OES), transmission electron microscopy (TEM, HRTEM), and differential scanning calorimetry (DSC). The nanoparticle size, chemical composition, and Cu-Ni nanoparticles melting temperature depression were obtained. The experimental temperatures of melting of nanoparticles were in good agreement with the theoretical CALPHAD predictions considering surface energy.Fázový diagram nanoslitiny Cu-Ni respektující velikost nanočástic jako další proměnné byl vypočten metodou CALPHAD. Vzorky Cu-Ni nanoslitin byly připraveny solvotermální syntézou z prekurzorů kovů. Tyto vzorky byly charakterizovány pomocí dynamického rozptylu světla (DLS), infračervené spektroskopie (IR) s indukčně vázanou plazmou a optickou emisní spektroskopií (ICP / OES), transmisní elektronovou mikroskopií (TEM, HRTEM) a diferenciální skenovací kalorimetrií (DSC). Velikost nanočástic, chemické složení a Cu-Ni deprese teploty tání nanočástic byly získány experimentálně a v dobré shodě s teoretickou předpovědí metodou CALPHAD s uvážením povrchové energie nanočástic

The Group B Streptococcal surface antigen I/II protein, BspC, interacts with host vimentin to promote adherence to brain endothelium and inflammation during the pathogenesis of meningitis

Streptococcus agalactiae (Group B Streptococcus, GBS) normally colonizes healthy adults but can cause invasive disease, such as meningitis, in the newborn. To gain access to the central nervous system, GBS must interact with and penetrate brain or meningeal blood vessels; however, the exact mechanisms are still being elucidated. Here, we investigate the contribution of BspC, an antigen I/II family adhesin, to the pathogenesis of GBS meningitis. Disruption of the bspC gene reduced GBS adherence to human cerebral microvascular endothelial cells (hCMEC), while heterologous expression of BspC in non-adherent Lactococcus lactis conferred bacterial attachment. In a murine model of hematogenous meningitis, mice infected with ΔbspC mutants exhibited lower mortality as well as decreased brain bacterial counts and inflammatory infiltrate compared to mice infected with WT GBS strains. Further, BspC was both necessary and sufficient to induce neutrophil chemokine expression. We determined that BspC interacts with the host cytoskeleton component vimentin and confirmed this interaction using a bacterial two-hybrid assay, microscale thermophoresis, immunofluorescent staining, and imaging flow cytometry. Vimentin null mice were protected from WT GBS infection and also exhibited less inflammatory cytokine production in brain tissue. These results suggest that BspC and the vimentin interaction is critical for the pathogenesis of GBS meningitis

Role of picornavirus 2A protease in inhibition of host nucleo-cytoplasmic transport /by Nogi Park.

Rhinovirus and poliovirus contain positive stranded RNA as their genome and perform viral translation and replication in the host cytoplasm. Previous studies showed that certain host nuclear factors interact with the viral genome and proteins in the cytoplasm. Consistent with cytoplasmic accumulations of host nuclear factors, the inhibition of certain host nuclear import pathways during infection was demonstrated along with the cleavage of nuclear pore complex (NPC) proteins such as Nup153, Nup98 and Nup62. These Nups contain phenylalanine-glycine (FG) rich regions that play an important role in nuclear transport.;The cleavage mechanism of Nups and the contribution of Nup cleavage in the inhibition of nuclear import during infection have not been explained clearly. I hypothesized that viral proteases specifically targeted Nups to inhibit nuclear import during infection. To test this hypothesis, this project has pursued the following questions; (1) Are viral proteases responsible for the Nup cleavage? (2) Where are the cleavage sites in target Nups? (3) What is the consequence of Nup cleavage on NPC composition during viral infection?;To test the contribution of viral protease in Nup cleavage, uninfected HeLa whole cell lysates and purified Nup62 and Nup98 were incubated with purified viral proteases, 2Apro and 3Cpro . These assays show that 2A pro plays a major role in the cleavage of Nup62 and Nup98.2Apro cleavage sites in Nup62 and Nup98 were identified by a combination of amino-terminal sequencing of cleavage products and sequence analysis and confirmed by in vitro cleavage assay with in vitro translated mutant Nups. Identified cleavage sites in both Nup62 and Nup98 were located in between the N-terminal FG rich region and C-terminal region. Immunofluorescence assays with mock- and virus-infected HeLa cells shows that the N-terminal FG rich regions of Nup62 and Nup98 were released from the NPC during infection.;Based on these results, this study suggests that poliovirus and rhinovirus 2Apro specifically targets the FG regions of Nups to inhibit normal host nuclear transport.Thesis (Ph. D., Microbiology, Molecular Biology and Biochemistry)--University of Idaho, August 2009