Antibody-mediated immunity to the obligate intracellular bacterial pathogen Coxiella burnetii is Fc receptor- and complement-independent

<p>Abstract</p> <p>Background</p> <p>The obligate intracellular bacterial pathogen <it>Coxiella burnetii </it>causes the zoonosis Q fever. The intracellular niche of <it>C. burnetii </it>has led to the assumption that cell-mediated immunity is the most important immune component for protection against this pathogen. However, passive immunization with immune serum can protect naïve animals from challenge with virulent <it>C. burnetii</it>, indicating a role for antibody (Ab) in protection. The mechanism of this Ab-mediated protection is unknown. Therefore, we conducted a study to determine whether Fc receptors (FcR) or complement contribute to Ab-mediated immunity (AMI) to <it>C. burnetii</it>.</p> <p>Results</p> <p>Virulent <it>C. burnetii </it>infects and replicates within human dendritic cells (DC) without inducing their maturation or activation. We investigated the effects of Ab opsonized <it>C. burnetii </it>on human monocyte-derived and murine bone marrow-derived DC. Infection of DC with Ab-opsonized <it>C. burnetii </it>resulted in increased expression of maturation markers and inflammatory cytokine production. Bacteria that had been incubated with naïve serum had minimal effect on DC, similar to virulent <it>C. burnetii </it>alone. The effect of Ab opsonized <it>C. burnetii </it>on DC was FcR dependent as evidenced by a reduced response of DC from FcR knockout (FcR k/o) compared to C57Bl/6 (B6) mice. To address the potential role of FcR in Ab-mediated protection in vivo, we compared the response of passively immunized FcR k/o mice to the B6 controls. Interestingly, we found that FcR are not essential for AMI to <it>C. burnetii </it>in vivo. We subsequently examined the role of complement in AMI by passively immunizing and challenging several different strains of complement-deficient mice and found that AMI to <it>C. burnetii </it>is also complement-independent.</p> <p>Conclusion</p> <p>Despite our data showing FcR-dependent stimulation of DC in vitro, Ab-mediated immunity to <it>C. burnetii </it>in vivo is FcR-independent. We also found that passive immunity to this pathogen is independent of complement.</p

Experimental and modelling evidence for structural crossover in supercritical CO₂

Physics of supercritical state is understood to a much lesser degree compared to subcritical liquids. Carbon dioxide in particular has been intensely studied, yet little is known about the supercritical part of its phase diagram. Here, we combine neutron scattering experiments and molecular dynamics simulations and demonstrate the structural crossover at the Frenkel line. The crossover is seen at pressures as high as 14 times the critical pressure and is evidenced by changes of the main features of the structure factor and pair distribution functions

Liquid flyback booster pre-phase: A study assessment

The concept of a flyback booster has been around since early in the shuttle program. The original two-stage shuttle concepts used a manned flyback booster. These boosters were eliminated from the program for funding and size reasons. The current shuttle uses two Redesigned Solid Rocket Motors (RSRM's), which are recovered and refurbished after each flight; this is one of the major cost factors of the program. Replacement options have been studied over the past ten years. The conclusion reached by the most recent study is that the liquid flyback booster (LFBB) is the only competitive option from a life-cycle cost perspective. The purpose of this study was to assess the feasibility and practicality of LFBB's. The study provides an expansion of the recommendations made during the aforementioned study. The primary benefits are the potential for enhanced reusability and a reduction of recurring costs. The potential savings in vehicle turnaround could offset the up-front costs. Development of LFBB's requires a commitment to the shuttle program for 20 to 30 years. LFBB's also offer enhanced safety and abort capabilities. Currently, any failure of an RSRM can be considered catastrophic, since there are no intact abort capabilities during the burn of the RSRM's. The performance goal of the LFBB's was to lift a fully loaded orbiter under optimal conditions, so as not to be the limiting factor of the performance capability of the shuttle. In addition, a final benefit is the availability of growth paths for applications other than shuttle

"You're actually part of the team" : a qualitative study of a novel transitional role from medical student to doctor

Background Optimizing transitions from final year of medical school and into first post graduate year has important implications for students, patients and the health care system. Student experiences during novel transitional roles can provide insights into potential opportunities for final year curricula. We explored the experiences of medical students in a novel transitional role and their ability to continue learning whilst working as part of a medical team. Methods Novel transitional role for final year medical students were created in partnership by medical schools and state health departments in 2020 in response to the COVID-19 pandemic and the need for a medical surge workforce. Final year medical students from an undergraduate entry medical school were employed as Assistants in Medicine (AiMs) in urban and regional hospitals. A qualitative study with semi-structured interviews at two time points was used to obtain experiences of the role from 26 AiMs. Transcripts were analyzed using deductive thematic analysis with Activity theory as a conceptual lens. Results This unique role was defined by the objective of supporting the hospital team. Experiential learning opportunities in patient management were optimized when AiMs had opportunities to contribute meaningfully. Team structure and access to the key instrument, the electronic medical record, enabled participants to contribute meaningfully, whilst contractual arrangements and payments formalized the obligations to contribute. Conclusions The experiential nature of the role was facilitated by organizational factors. Structuring teams to involve a dedicated medical assistant position with specific duties and access to the electronic medical record sufficient to complete duties are key to successful transitional roles. Both should be considered when designing transitional roles as placements for final year medical students

A mouse model for MERS coronavirus-induced acute respiratory distress syndrome

Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel virus that emerged in 2012, causing acute respiratory distress syndrome (ARDS), severe pneumonia-like symptoms, and multi-organ failure, with a case fatality rate of ~36%. Limited clinical studies indicate that humans infected with MERS-CoV exhibited pathology consistent with late stages of ARDS, which is reminiscent of disease observed in patients infected with SARS coronavirus. Models of MERS-CoV-induced severe respiratory disease have been difficult to achieve, and small animal models traditionally used to investigate viral pathogenesis (mouse, hamster, guinea pig, and ferret) are naturally resistant to MERS-CoV. Therefore, we used CRISPR/Cas9 to modify the mouse genome to encode two human amino acids (288 and 330) in the dipeptidyl peptidase 4 receptor, making mice susceptible to MERS-CoV replication. Serial MERS-CoV passage in these engineered mice was then used to generate a mouse-adapted virus that replicated efficiently within the lungs, and evoked symptoms indicative of severe acute respiratory distress syndrome (ARDS), including decreased survival, extreme weight loss, decreased pulmonary function, pulmonary hemorrhage, and pathological signs indicative of end stage lung disease. Importantly, therapeutic countermeasures comprising MERS-CoV neutralizing antibody treatment or a MERS-CoV spike protein vaccine protected engineered mice against MERS-CoV-induced ARDS

Liquid Flyback Booster Pre-Phase A Study Assessment

Mw concept of a flyback booster has been around since early in the Shuttle program. The original two-stage Shuttle concepts used a manned flyback booster. These boosters were eliminated from the program for funding and size reasons. The current Shuttle uses two Redesigned Solid Rocket Motors (RSRMs), which are recovered and refurbished after each flight; this is one of the major cost factors of the program. Replacement options have been studied over the past ten years. The conclusion reached by the most recent study is that the liquid flyback booster (LFBB) is the only competitive option from a life-cycle cost perspective. The purpose of this study was to assess the feasibility and practicality of LFBBs. The study provides an expansion of the recommendations made during the during the aforementioned study. The primary benefits are the potential for enhanced reusability and a reuction of recurring costs. The potential savings in vehicle turnaround could offset the up-front costs. Development of LFBBs requires a commitment to the Shuttle program for 20 to 30 years. LFBBs also offer enhanced safety and abort capabilities. Currently, any failure of an RSRM can be considered catastrophic since them we no intact abort capabilities during the burn of the RSRMS. The performance goal of the LFBBs was to lift a fully loaded Orbiter under optimal conditions, so as not to be the limiting factor of the performance capability of the Shuttle. In addition, a final benefit is the availability of growth paths for applications other than the Shuttle

Highly efficient separation of actinides from lanthanides by a phenanthroline-derived bis-triazine ligand

The synthesis, lanthanide complexation, and solvent ex- traction of actinide(III) and lanthanide(III) radiotracers from nitric acid solutions by a phenanthroline-derived quadridentate bis-triazine ligand are described. The ligand separates Am(III) and Cm(III) from the lanthanides with remarkably high efficiency, high selectivity, and fast extraction kinetics compared to its 2,2'-bipyridine counterpart. Structures of the 1:2 bis-complexes of the ligand with Eu(III) and Yb(III) were elucidated by X-ray crystallography and force field calculations, respec-tively. The Eu(III) bis-complex is the first 1:2 bis-complex of a quadridentate bis-triazine ligand to be characterized by crystallography. The faster rates of extraction were verified by kinetics measurements using the rotating membrane cell technique in several diluents. The improved kinetics of metal ion extraction are related to the higher surface activity of the ligand at the phase interface. The improvement in the ligand's properties on replacing the bipyridine unit with a phenanthroline unit far exceeds what was anticipated based on ligand design alone

Comparative validation of standard, picture-sort and meal-based food-frequency questionnaires adapted for an elderly population of low socio-economic status

OBJECTIVE: To compare the validity of a modified Block food-frequency questionnaire (FFQ), a picture-sort administration of the FFQ (PSFFQ) and a meal pattern-based questionnaire (MPQ) in a multi-ethnic population of low socio-economic status (SES). DESIGN: Participants completed six 24-hour dietary recalls (24HR) over six months; the FFQ, PSFFQ and MPQ were completed in random order in the subsequent month. Instruments were interviewer-administered. The PSFFQ and MPQ were developed in formative research concerning difficulties for older adults in responding to standard food-frequency instruments. SETTING: Rural North Carolina, USA. Subjects One hundred and twenty-two African American, Native American and white adults aged > or = 65 years, with approximately one-third in each ethnic group. Inclusion criteria included education < or = 12 years and income < or = 150% of national poverty level or Medicaid recipient. RESULTS: Comparing median intakes from the average of the 24HR with the three diet assessment instruments, the MPQ tended to overestimate intakes compared with the FFQ and PSFFQ. Correlations among nutrients obtained by the 24HR and the other three instruments were generally statistically significant and positive. Across nutrients, the PSFFQ was most highly correlated with the 24HR for women, while the FFQ was most highly correlated with the 24HR for men. CONCLUSIONS: Dietary assessments using 24HR and FFQ were similar to results reported elsewhere, although correlations between 24HR and FFQ were somewhat lower. Interviewer-administered dietary assessments should be used with caution to evaluate dietary intake among older adults with low SES. Gender differences and the lower correlations should be investigated more thoroughly to assist in choosing dietary assessment instruments for this population

A “Coiled-Coil” Motif Is Important for Oligomerization and DNA Binding Properties of Human Cytomegalovirus Protein UL77

Human cytomegalovirus (HCMV) UL77 gene encodes the essential protein UL77, its function is characterized in the present study. Immunoprecipitation identified monomeric and oligomeric pUL77 in HCMV infected cells. Immunostaining of purified virions and subviral fractions showed that pUL77 is a structural protein associated with capsids. In silico analysis revealed the presence of a coiled-coil motif (CCM) at the N-terminus of pUL77. Chemical cross-linking of either wild-type pUL77 or CCM deletion mutant (pUL77ΔCCM) implicated that CCM is critical for oligomerization of pUL77. Furthermore, co-immunoprecipitations of infected and transfected cells demonstrated that pUL77 interacts with the capsid-associated DNA packaging motor components, pUL56 and pUL104, as well as the major capsid protein. The ability of pUL77 to bind dsDNA was shown by an in vitro assay. Binding to certain DNA was further confirmed by an assay using biotinylated 36-, 250-, 500-, 1000-meric dsDNA and 966-meric HCMV-specific dsDNA designed for this study. The binding efficiency (BE) was determined by image processing program defining values above 1.0 as positive. While the BE of the pUL56 binding to the 36-mer bio-pac1 containing a packaging signal was 10.0±0.63, the one for pUL77 was only 0.2±0.03. In contrast to this observation the BE of pUL77 binding to bio-500 bp or bio-1000 bp was 2.2±0.41 and 4.9±0.71, respectively. By using pUL77ΔCCM it was demonstrated that this protein could not bind to dsDNA. These data indicated that pUL77 (i) could form homodimers, (ii) CCM of pUL77 is crucial for oligomerization and (iii) could bind to dsDNA in a sequence independent manner