Efficient phagocytosis of Klebsiella pneumoniae strains that poorly bind to human polymorphonuclear leukocytes

The phagocytosis process of unencapsulated MIAT-negative strains that, although binding very poorly to human polymorphonuclear leukocytes (PMN) at 4°C, are efficiently killed by these cells at 37°C, was studied. At 37°C the number of bacteria bound to the PMN external surface was similar to that observed at 4°C (about 100 bacteria/100 PMN after 60 min); on the contrary the number of internalized bacteria was much higher (from 500 bacteria/100 PMN after 60 min). Interactions between phagocytosis-sensitive Klebsiella pneumoniae strains (PSK) and PMN were then compared with those of two isogenic Escherichia coli strains with and without type 1 fimbriae. Whereas PSK strain binding to blocked PMN was very slow and became significant only after 5–6 h, that of phagocytosis-sensitive fimbriated E. coli was rapid and efficient. Phagocytosis-resistant, non fimbriated E. coli strain bound with an efficiency that, within the first 60 min, was not very different from that of the PSK strains. However, longer incubations led to increases in PSK binding, whereas unfimbriated E. coli remained constant. PSK and fimbriated E. coli strains were efficiently internalized and killed, whereas the unfimbriated E. coli strain was not. It is suggested that PMN can phagocytize unopsonized bacteria through two different mechanisms. By one mechanism, observed with the fimbriated E. coli strain, PMN bind many more bacteria than those they can internalize. By the other, observed with PSK strains, PMN bind only the bacteria they can immediately internalize

Specific Lung Mucosal and Systemic Immune-Responses After Oral Immunization of Mice with Salmonella-Typhimurium-Aroa, Salmonella-Typhi Ty21a, and Invasive Escherichia-Coli Expressing Recombinant Pertussis Toxin S1 Subunit

Pertussis toxin (PT) is considered an essential protective component for incorporation into new generation vaccines against Bordetella pertussis, the causative agent of whooping cough. Traditionally, antipertussis vaccination has employed an intramuscular route. An alternative to this approach is to stimulate mucosal and systemic immune responses by oral immunization with live vaccine carrier strains of Salmonella spp. or Escherichia coli. Recombinant S1 subunit of pertussis toxin was expressed in the attenuated aroA mutant of Salmonella typhimurium, SL3261, in the human typhoid vaccine strain Salmonella typhi Ty21a, and in E. coli CAG629 containing the Shigella flexneri plasmid pWR110, which encodes bacterial invasiveness of epithelial cells. Expression of recombinant PT S1 subunit (rPT-S1) did not affect in vitro invasiveness of the tested strains, which retained the ability to adhere to and invade the embryonic human intestinal cell line HI-407. Following oral immunization of mice with the live vaccine strains expressing rPT-S1, immunoglobulin G (IgG), IgA, and IgM responses were monitored. IgG specific to PT was detected in serum samples of mice, while IgG and IgA specific to PT were detected in lung washes after oral immunization with living Salmonella spp. or E. coli (pWR110) expressing rPT-S1. Utilization of live oral vaccines expressing B. pertussis antigens, which stimulate both a systemic and lung mucosal response, may provide an attractive alternative to purified component vaccines against whooping cough

Direct Expression of Bordetella-Pertussis Filamentous Hemagglutinin in Escherichia-Coli and Salmonella-Typhimurium Aroa

Nonfused (i.e., nonhybrid) filamentous hemagglutinin (FHA) of Bordetella pertussis was efficiently expressed in Escherichia coli K-12 and Salmonella typhimurium aroA at levels higher than those found in wild-type B. pertussis when the upstream signals of the gene were replaced and the translation initiation region was engineered to optimize translational efficiency. Inclusion of part of the C-terminal FHA open reading frame, whose translation product does not appear to be part of the major secreted species of FHA, was shown to be important in achieving protein expression in both E. coli and S. typhimurium aroA; removal of the downstream gene sequence abolished recombinant FHA production. The levels of expression observed varied widely according to the construct and host bacterium used

The Toll-like Receptor 2/6 Ligand MALP-2 Reduces the Viability of Mycobacterium tuberculosis in Murine Macrophages

Toll-like receptors (TLRs) sense conserved structures of pathogens and influence macrophage functions. Here we investigated the impact of TLR signaling on the modulation of macrophage defense mechanisms against infection of Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis. We found that a synthetic derivative of the TLR2/6 agonist MALP-2 and the potent TLR4 agonist lipopolysaccharide inhibited the intracellular growth of MTB in murine macrophages. Likely the microbicidal effect was mediated by production of nitric oxide while it is still unclear the role played by release of TNF-α , IL-6, MIP-1β and IL-10. These results suggest that the activation of microbicidal defense via TLR ligands is an appealing target for the establishment on immune intervention against tuberculosis

Symmetry-projected wave functions in quantum Monte Carlo calculations

We consider symmetry-projected Hartree-Fock trial wave functions in constrained-path Monte Carlo (CPMC) calculations. Previous CPMC calculations have mostly employed Hartree-Fock (HF) trial wave functions, restricted or unrestricted. The symmetry-projected HF approach results in a hierarchy of wave functions with increasing quality: the more symmetries that are broken and restored in a self-consistent manner, the higher the quality of the trial wave function. This hierarchy is approximately maintained in CPMC calculations: the accuracy in the energy increases and the statistical variance decreases when further symmetries are broken and restored. Significant improvement is achieved in CPMC with the best symmetry-projected trial wave functions over those from simple HF. We analyze and quantify the behavior using the two-dimensional repulsive Hubbard model as an example. In the sign-problem-free region, where CPMC can be made exact but a constraint is deliberately imposed here, spin-projected wave functions remove the constraint bias. Away from half filling, spatial symmetry restoration in addition to that of the spin leads to highly accurate results from CPMC. Since the computational cost of symmetry-projected HF trial wave functions in CPMC can be made to scale algebraically with system size, this provides a potentially general approach for accurate calculations in many-fermion systems

Minimally Invasive Laminate Veneer Therapy for Maxillary Central Incisors.

Minimally invasive dentistry is a considered process that requires the clinician to be prepared with the ideal sequence and the tools needed. This report describes a well-planned ultraconservative approach using only two ceramic laminate veneers for the maxillary central incisors to significantly improve the patient's overall smile. A 30-year-old female presented with the chief complaints of having diastemas between the central and lateral incisors as well as incisal wear. Diagnostic wax-up and mock-up were performed, and the patient approved the minimally invasive treatment with veneers only for central incisors. A reduction guide aided the conservative tooth preparations, and hand-crafted feldspathic veneers were bonded under total isolation with a rubber dam. The two final conservative veneers significantly improved the smile and fulfilled the patient's expectations. Following proper planning and sequencing, predictable outcomes were obtained and fulfilled the patient's esthetic demands. Minimally invasive restorative dentistry with only two single veneers can impact the entire smile frame. Overtreatment in the esthetic zone is unnecessary to meet a patient's esthetic expectations

Compression Behavior of Fluted-Core Composite Panels

In recent years, fiber-reinforced composites have become more accepted for aerospace applications. Specifically, during NASA s recent efforts to develop new launch vehicles, composite materials were considered and baselined for a number of structures. Because of mass and stiffness requirements, sandwich composites are often selected for many applications. However, there are a number of manufacturing and in-service concerns associated with traditional honeycomb-core sandwich composites that in certain instances may be alleviated through the use of other core materials or construction methods. Fluted-core, which consists of integral angled web members with structural radius fillers spaced between laminate face sheets, is one such construction alternative and is considered herein. Two different fluted-core designs were considered: a subscale design and a full-scale design sized for a heavy-lift-launch-vehicle interstage. In particular, axial compression of fluted-core composites was evaluated with experiments and finite-element analyses (FEA); axial compression is the primary loading condition in dry launch-vehicle barrel sections. Detailed finite-element models were developed to represent all components of the fluted-core construction, and geometrically nonlinear analyses were conducted to predict both buckling and material failures. Good agreement was obtained between test data and analyses, for both local buckling and ultimate material failure. Though the local buckling events are not catastrophic, the resulting deformations contribute to material failures. Consequently, an important observation is that the material failure loads and modes would not be captured by either linear analyses or nonlinear smeared-shell analyses. Compression-after-impact (CAI) performance of fluted core composites was also investigated by experimentally testing samples impacted with 6 ft.-lb. impact energies. It was found that such impacts reduced the ultimate load carrying capability by approximately 40% on the subscale test articles and by less than 20% on the full-scale test articles. Nondestructive inspection of the damage zones indicated that the detectable damage was limited to no more than one flute on either side of any given impact. More study is needed, but this may indicate that an inherent damage-arrest capability of fluted core could provide benefits over traditional sandwich designs in certain weight-critical applications