Functional studies of serine/threonine protein kinase signalling in Mycobacterium tuberculosis.

Fork-head associated (FHA) domains function in the assembly of signalling complexes through specific interactions with phospho-threonine motifs. FHA domains are found in eukaryotes, and intriguingly in a subset of bacterial species including Mycobacteria tuberculosis. In addition, the M. tuberculosis genome encodes 11 eukaryotic-like serine/threonine protein kinases (STPKs). In M. tuberculosis, the co-expression of FHA domain-containing proteins and STPKs strongly suggest that these bacterial FHA domain-containing proteins engage in phospho-dependent protein-protein interaction, and FHA dependent processes in bacteria are controlled by STPK-dependent phosphorylation. This study describes a body of biophysical and biochemical experiments which characterises the interactions between two STPKs, PknA and PknB, and three FHA domain-containing proteins, Rv0019, Rv0020 and Rv1827. Isothermal titration calorimetry and surface plasmon resonance combined with site-directed mutagenesis are used to investigate interactions of kinase domains with FHA domains. These experiments reveal that the FHA domains interact with specific phospho-threonine residues located within the kinase domain activation loop. Additionally, in vitro kinase assays demonstrated that the interactions also involve phosphorylation of the FHA domains and/or adjacent segments of the protein. These data suggest that STPK-mediated signalling in M. tuberculosis involves a complex series of interactions and suggests a 'molecular docking' model for the phosphorylation of FHA domain-containing proteins. Finally, high-throughput assays have been used to identify several inhibitors of PknB which will serve as the basis for development of novel antimicrobial therapies

Accounting B

Exam paper for second semeste

Restriction of memory B cell differentiation at the germinal center B cell positive selection stage

Memory B cells (MBCs) are key for protection from reinfection. However, it is mechanistically unclear how germinal center (GC) B cells differentiate into MBCs. MYC is transiently induced in cells fated for GC expansion and plasma cell (PC) formation, so-called positively selected GC B cells. We found that these cells coexpressed MYC and MIZ1 (MYC-interacting zinc-finger protein 1 [ZBTB17]). MYC and MIZ1 are transcriptional activators; however, they form a transcriptional repressor complex that represses MIZ1 target genes. Mice lacking MYC-MIZ1 complexes displayed impaired cell cycle entry of positively selected GC B cells and reduced GC B cell expansion and PC formation. Notably, absence of MYC-MIZ1 complexes in positively selected GC B cells led to a gene expression profile alike that of MBCs and increased MBC differentiation. Thus, at the GC positive selection stage, MYC-MIZ1 complexes are required for effective GC expansion and PC formation and to restrict MBC differentiation. We propose that MYC and MIZ1 form a module that regulates GC B cell fate

Neighborhood disadvantage and immune-related illnesses among residents living in the US Gulf States

Purpose: Neighborhood disadvantage has been associated with increased risk for pneumonia and influenza-associated hospitalizations. Few studies, however, have investigated how neighborhood disadvantage may influence immune-related illnesses. The aim of this study was to examine the association between neighborhooddisadvantage and immune-related illnesses. Methods: We used data from the Gulf Long-term Follow-up (GuLF) Study (n = 32,608). Our analytic sample included home visit participants (n = 11,193) who had complete information on exposure and covariates (n = 10,543). Neighborhood disadvantage was assessed using the 2013 Area Deprivation Index (ADI), which assigns a ranking of 1 to 100 for lowest to highest disadvantage. We linked ADI to participants’ geocoded enrollment addresses at the census block group level. ADI was categorized into quartiles based on the national distribution with the first quartile as the referent. Immune-related illnesses self-reported at the home visit (May 2011–May 2013) included occurrence of shingles, pneumonia, cold sores, flu, and colds since the Deepwater Horizon oil spill (April 2010). Frequent colds and frequent flu were defined as ≥4 colds and ≥2 episodes of flu since the spill. An aggregated outcome, based on occurrence of any pneumonia, cold sores, flu, and ≥4 colds since the spill, was also examined. We assessed the association of each outcome with ADI using multivariable log-binomial regression adjusting for individual-level demographics, behavioral factors, kids at home, and season of interview completion. Results: We found elevated prevalence ratios (PR) and 95% confidence intervals (CI) for pneumonia associated with ADI in the third (PR: 2.04, 95% CI: 1.04, 4.02) and fourth (PR: 2.00; 95% CI: 1.00, 3.98) quartiles. PRs for frequent colds were also elevated for increasing ADI quartiles, but with confidence intervals including the null value. Conclusions: The observed associations of frequent colds and pneumonia with increasing neighborhood disadvantage may warrant further research on this topi

Height and risk of death among men and women: aetiological implications of associations with cardiorespiratory disease and cancer mortality

OBJECTIVES: Height is inversely associated with cardiovascular disease mortality risk and has shown variable associations with cancer incidence and mortality. The interpretation of findings from previous studies has been constrained by data limitations. Associations between height and specific causes of death were investigated in a large general population cohort of men and women from the West of Scotland. DESIGN: Prospective observational study. SETTING: Renfrew and Paisley, in the West of Scotland. SUBJECTS: 7052 men and 8354 women aged 45-64 were recruited into a study in Renfrew and Paisley, in the West of Scotland, between 1972 and 1976. Detailed assessments of cardiovascular disease risk factors, morbidity and socioeconomic circumstances were made at baseline. MAIN OUTCOME MEASURES: Deaths during 20 years of follow up classified into specific causes. RESULTS: Over the follow up period 3347 men and 2638 women died. Height is inversely associated with all cause, coronary heart disease, stroke, and respiratory disease mortality among men and women. Adjustment for socioeconomic position and cardiovascular risk factors had little influence on these associations. Height is strongly associated with forced expiratory volume in one second (FEV1) and adjustment for FEV1 considerably attenuated the association between height and cardiorespiratory mortality. Smoking related cancer mortality is not associated with height. The risk of deaths from cancer unrelated to smoking tended to increase with height, particularly for haematopoietic, colorectal and prostate cancers. Stomach cancer mortality was inversely associated with height. Adjustment for socioeconomic position had little influence on these associations. CONCLUSION: Height serves partly as an indicator of socioeconomic circumstances and nutritional status in childhood and this may underlie the inverse associations between height and adulthood cardiorespiratory mortality. Much of the association between height and cardiorespiratory mortality was accounted for by lung function, which is also partly determined by exposures acting in childhood. The inverse association between height and stomach cancer mortality probably reflects Helicobacter pylori infection in childhood resulting inor being associated withshorter height. The positive associations between height and several cancers unrelated to smoking could reflect the influence of calorie intake during childhood on the risk of these cancers

Co-activation of NF-κB and MYC renders cancer cells addicted to IL6 for survival and phenotypic stability

NF-κB and MYC are found co-deregulated in human B and plasma-cell cancers. In physiology, NF-κB is necessary for terminal B-to-plasma cell differentiation, whereas MYC repression is required. It is thus unclear if NF-κB/MYC co-deregulation is developmentally compatible in carcinogenesis and/or impacts cancer cell differentiation state, possibly uncovering unique sensitivities. Using a mouse system to trace cell lineage and oncogene activation we found that NF-κB/MYC co-deregulation originated cancers with a plasmablast-like phenotype, alike human plasmablastic-lymphoma and was linked to t(8;14)[MYC-IGH] multiple myeloma. Notably, in contrast to NF-κB or MYC activation alone, co-deregulation rendered cells addicted to IL6 for survival and phenotypic stability. We propose that conflicting oncogene-driven differentiation pressures can be accommodated at a cost in poorly-differentiated cancers. SIGNIFICANCE: Our studies improve the understanding of cancer pathogenesis by demonstrating that co-deregulation of NF-κB and MYC synergize in forming a cancer with a poorly-differentiated state. The cancers in the mouse system share features with human Plasmablastic lymphoma that has a dismal prognosis and no standard of care, and with t(8;14)[MYC-IGH] Multiple myeloma, which is in overall resistant to standard therapy. Notably, we found that NF-κB and MYC co-deregulation uniquely render cells sensitive to IL6 deprivation, providing a road-map for patient selection. Because of the similarity of the cancers arising in the compound mutant mouse model with that of human Plasmablastic lymphoma and t(8;14)[MYC-IGH] Multiple myeloma, this model could serve in preclinical testing to investigate novel therapies for these hard-to-treat diseases

Variability of the pullout strength of cancellous bone screws with cement augmentation

Background Orthopaedic surgeons often face clinical situations where improved screw holding power in cancellous bone is needed. Injectable calcium phosphate cements are one option to enhance fixation. Methods Paired screw pullout tests were undertaken in which human cadaver bone was augmented with calcium phosphate cement. A finite element model was used to investigate sensitivity to screw positional placement. Findings Statistical analysis of the data concluded that the pullout strength was generally increased by cement augmentation in the in vitro human cadaver tests. However, when comparing the individual paired samples there were surprising results with lower strength than anticipated after augmentation, in apparent contradiction to the generally expected conclusion. Investigation using the finite element model showed that these strength reductions could be accounted for by small screw positional changes. A change of 0.5 mm might result in predicted pullout force changes of up to 28%. Interpretation Small changes in screw position might lead to significant changes in pullout strength sufficient to explain the lower than expected individual pullout values in augmented cancellous bone. Consequently whilst the addition of cement at a position of low strength would increase the pullout strength at that point, it might not reach the pullout strength of the un-augmented paired test site. However, the overall effect of cement augmentation produces a significant improvement at whatever point in the bone the screw is placed. The use of polymeric bone-substitute materials for tests may not reveal the natural variation encountered in tests using real bone structures.Dr V. Stadelmann (AOR, Davos, Switzerland) and Mr. M. Behrens (Stryker, Selzach, Switzerland). Professor Procter and Dr Arnoldi were employed by Stryker Trauma. Dr Bennani's PhD studies at Brunel University were funded by Stryker Trauma AG

Forward pi^0 Production and Associated Transverse Energy Flow in Deep-Inelastic Scattering at HERA

Deep-inelastic positron-proton interactions at low values of Bjorken-x down to x \approx 4.10^-5 which give rise to high transverse momentum pi^0 mesons are studied with the H1 experiment at HERA. The inclusive cross section for pi^0 mesons produced at small angles with respect to the proton remnant (the forward region) is presented as a function of the transverse momentum and energy of the pi^0 and of the four-momentum transfer Q^2 and Bjorken-x. Measurements are also presented of the transverse energy flow in events containing a forward pi^0 meson. Hadronic final state calculations based on QCD models implementing different parton evolution schemes are confronted with the data.Comment: 27 pages, 8 figures and 3 table

Low Q^2 Jet Production at HERA and Virtual Photon Structure

The transition between photoproduction and deep-inelastic scattering is investigated in jet production at the HERA ep collider, using data collected by the H1 experiment. Measurements of the differential inclusive jet cross-sections dsigep/dEt* and dsigmep/deta*, where Et* and eta* are the transverse energy and the pseudorapidity of the jets in the virtual photon-proton centre of mass frame, are presented for 0 < Q2 < 49 GeV2 and 0.3 < y < 0.6. The interpretation of the results in terms of the structure of the virtual photon is discussed. The data are best described by QCD calculations which include a partonic structure of the virtual photon that evolves with Q2.Comment: 20 pages, 5 Figure