The Polysaccharide Capsule of Streptococcus pneumonia Partially Impedes MyD88-Mediated Immunity during Pneumonia in Mice

Toll-like receptors (TLR) and the downstream adaptor protein MyD88 are considered crucial for protective immunity during bacterial infections. Streptococcus (S.) pneumoniae is a human respiratory pathogen and a large majority of clinical pneumococcal isolates expresses an external polysaccharide capsule. We here sought to determine the role of pneumococcal capsule in MyD88-mediated antibacterial defense during S. pneumonia pneumonia. Wild type (WT) and Myd88^-/- mice were inoculated intranasally with serotype 2 S. pneumoniae D39 or with an isogenic capsule locus deletion mutant (D39∆cps), and analysed for bacterial outgrowth and inflammatory responses in the lung. As compared to WT mice, Myd88^-/- mice infected with D39 demonstrated a modestly impaired bacterial clearance accompanied by decreased inflammatory responses in the lung. Strikingly, while WT mice rapidly cleared D39∆cps, Myd88^-/- mice showed 10⁵-fold higher bacterial burdens in their lungs and dissemination to blood 24 hours after infection. These data suggest that the pneumococcal capsule impairs recognition of TLR ligands expressed by S. pneumoniae and thereby partially impedes MyD88-mediated antibacterial defense.     

Induction of Strain-Transcending Antibodies Against Group A PfEMP1 Surface Antigens from Virulent Malaria Parasites

Sequence diversity in pathogen antigens is an obstacle to the development of interventions against many infectious diseases. In malaria caused by Plasmodium falciparum, the PfEMP1 family of variant surface antigens encoded by var genes are adhesion molecules that play a pivotal role in malaria pathogenesis and clinical disease. PfEMP1 is a major target of protective immunity, however, development of drugs or vaccines based on PfEMP1 is problematic due to extensive sequence diversity within the PfEMP1 family. Here we identified the PfEMP1 variants transcribed by P. falciparum strains selected for a virulence-associated adhesion phenotype (IgM-positive rosetting). The parasites transcribed a subset of Group A PfEMP1 variants characterised by an unusual PfEMP1 architecture and a distinct N-terminal domain (either DBLα1.5 or DBLα1.8 type). Antibodies raised in rabbits against the N-terminal domains showed functional activity (surface reactivity with live infected erythrocytes (IEs), rosette inhibition and induction of phagocytosis of IEs) down to low concentrations (<10 µg/ml of total IgG) against homologous parasites. Furthermore, the antibodies showed broad cross-reactivity against heterologous parasite strains with the same rosetting phenotype, including clinical isolates from four sub-Saharan African countries that showed surface reactivity with either DBLα1.5 antibodies (variant HB3var6) or DBLα1.8 antibodies (variant TM284var1). These data show that parasites with a virulence-associated adhesion phenotype share IE surface epitopes that can be targeted by strain-transcending antibodies to PfEMP1. The existence of shared surface epitopes amongst functionally similar disease-associated P. falciparum parasite isolates suggests that development of therapeutic interventions to prevent severe malaria is a realistic goal.     

Eimeria refractile body proteins contain two potentially functional characteristics: Transhydrogenase and carbohydrate transport

Abstract cDNA encoding an immunogenic protein from partially sporulated oocysts of Eimeria acervulina was cloned and used to search for the homologous counterpart in Eimeria tenella . Monospecific antibodies were raised against the recombinant expression product. Using these antibodies, the parasite proteins were found to be localised in the refractile bodies. The derived amino-acid sequences were compared by computer using the SWISSPROT protein database. In addition to high homology between the Eimeria species, extensive similarity was found with pyridine nucleotide transhydrogenase from Escherichia coli . Comparison with the sugar signature database also resulted in a possible sugar binding domain present only in the Eimeria proteins. It is possible that the corresponding parasite proteins play a role in the recently discovered mannitol cycle of Eimeria .     

New integrative method to generate Bacillus subtilis recombinant strains free of selection markers

The novel method described in this paper combines the use of blaI, which encodes a repressor involved in Bacillus licheniformis BlaP beta-lactamase regulation, an antibiotic resistance gene, and a B. subtilis strain (BS1541) that is conditionally auxotrophic for lysine. We constructed a BlaI cassette containing blaI and the spectinomycin resistance genes and two short direct repeat DNA sequences, one at each extremity of the cassette. The BS1541 strain was obtained by replacing the B. subtilis P(lysA) promoter with that of the P(blaP) beta-lactamase promoter. In the resulting strain, the cloning of the blaI repressor gene confers lysine auxotrophy to BS1541. After integration of the BlaI cassette into the chromosome of a conditionally lys-auxotrophic (BS1541) strain by homologous recombination and positive selection for spectinomycin resistance, the eviction of the BlaI cassette was achieved by single crossover between the two short direct repeat sequences. This strategy was successfully used to inactivate a single gene and to introduce a gene of interest in the Bacillus chromosome. In both cases the resulting strains are free of selection marker. This allows the use of the BlaI cassette to repeatedly further modify the Bacillus chromosome.     

A fast strong coupling algorithm for the partitioned fluid–structure interaction simulation of BMHVs

The numerical simulation of Bileaflet Mechanical Heart Valves (BMHVs) has gained strong interest in the last years, as a design and optimisation tool. In this paper, a strong coupling algorithm for the partitioned fluid–structure interaction simulation of a BMHV is presented. The convergence of the coupling iterations between the flow solver and the leaflet motion solver is accelerated by using the Jacobian with the derivatives of the pressure and viscous moments acting on the leaflets with respect to the leaflet accelerations. This Jacobian is numerically calculated from the coupling iterations. An error analysis is done to derive a criterion for the selection of useable coupling iterations. The algorithm is successfully tested for two 3D cases of a BMHV and a comparison is made with existing coupling schemes. It is observed that the developed coupling scheme outperforms these existing schemes in needed coupling iterations per time step and CPU time.     

Interaction of androgen response elements with the DNA-binding domain of the rat androgen receptor expressed in Escherichia coli

A fragment of the rat androgen receptor (amino acids 533-637) containing the DNA-binding domain was produced in Escherichia coli as a fusion product with protein A of Staphylococcus aureus. The fusion protein was purified on IgG-Sepharose, a method that does not involve the use of denaturing agents. Approximately 4 mg of fusion protein was obtained from 500 ml of bacterial culture. In gel shift assays, the recombinant DNA-binding domain displays an affinity for a fragment of the long terminal repeat of mouse mammary tumor virus and for an intronic fragment of the gene coding for the C3 component of the androgen-regulated rat prostatic binding protein. In a DNase I footprinting assay, the fusion protein protects a sequence in the C3 fragment that has previously been shown to act as a functional androgen response element. Interestingly, a single base pair mutation in the response element, which abolishes androgen inducibility, also destroys the ability to interact with the recombinant androgen receptor DNA-binding domain.     

Geographic,seasonal, and diurnal surface behavior of harbor porpoises

During ship surveys harbor porpoises are only visible when breaking the sea surface to breathe, while during aerial surveys they may be seen down to 2 m below the surface. The fractions of time spent at these two depths can be used for correcting visual surveys to actual population estimates, which are essential information on the status and management of the species. Thirty‐five free‐ranging harbor porpoises (Phocoena phocoena) were tracked in the region between the Baltic and the North Sea for 25–349 d using Argos satellite transmitters. No differences were found in surface behavior between geographical areas or the size of the animals. Slight differences were found between the two sexes and time of day. Surface time peaked in April, where 6% was spent with the transmitter above surface and 61.5% between 0 and 2 m depth, while the minimum values occurred in February (3.4% and 42.5%, respectively). The analyses reveal that individual variation among porpoises is the most important factor in explaining variation in surface rates. However, the large number of animals documented in the present study covering a wide range of age and sex groups justifies the use of the seasonal average surface times for correcting abundance surveys.     

Trait variation in juvenile plants from the soil seed bank of temperate forests in relation to macro- and microclimate

Aim

The soil seed bank is a key component of the biodiversity of plant communities, but various aspects of its functioning in temperate forest ecosystems are still unknown. We here adopted a trait-based approach to investigate the effects of macro- and microclimatic gradients on the juvenile plant communities from the realized seed bank of two types of European temperate forest.

Location

Oak-dominated forests in Italy and Belgium.

Methods

We analysed the variation of key functional traits (plant height, leaf area, leaf dry weight, specific leaf area and leaf number) of juvenile plants from the realised soil seed bank in relation to elevation (from 0 to 800 m a.s.l.), forest type (thinned and unthinned forest) and distance to the forest edge. We translocated soil samples from the forest core to the edge (and vice versa) and from high- to low-elevation forests to test the effects of edge and warming respectively.

Results

Taller communities developed at the forest edge due to higher light availability and warmer temperatures. The translocation from the core to the edge did not significantly modify mean trait values. Instead, the shadier and cooler microclimate of the forest core reduced the mean leaf area, mean dry weight, height and leaf number in the communities realised from the edge soil. The translocation from high- to lowland forests led to increased values for all traits (except specific leaf area). Edge vs core trait variation was more driven by intraspecific variability, whereas the translocation from high- to low-elevation forests caused trait changes mostly due to species turnover.

Conclusions

Global warming might result in a functional shift of the understorey due to both an early filtering effect on the seedlings from soil seed banks and their adaptive trait adjustments to temperature increase. Furthermore, our study underpins the importance of edge vs core microclimate in driving the functional composition of the realised soil seed bank.     

The single-cell epigenomic and transcriptional landscape of immunity to influenza vaccination

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