Comparisons of resistance of CF and Non-CF pathogens to Hydrogen Peroxide and Hypochlorous Acid Oxidants In Vitro

<p>Abstract</p> <p>Background</p> <p>Cystic fibrosis (CF) lung disease has a unique profile of pathogens predominated by <it>Pseudomonas aeruginosa </it>(PsA) and <it>Staphylococcus aureus </it>(SA). These microorganisms must overcome host immune defense to colonize the CF lungs. Polymorphonuclear neutrophils are a major component of the host defense against bacterial infection. A crucial microbicidal mechanism is the production of oxidants including hydrogen peroxide (H₂O₂) and hypochlorous acid (HOCl) by neutrophils to achieve efficient bacterial killing. To determine to what degrees various CF pathogens resist the oxidants relative to non-CF pathogens, we compared the susceptibility of PsA, SA, <it>Burkholderia cepacia </it>(BC), <it>Klebsiella pneumoniae </it>(KP), and <it>Escherichia coli </it>(EC) to various concentrations of H₂O₂or HOCl, <it>in vitro</it>. The comparative oxidant-resistant profiles were established. Oxidant-induced damage to ATP production and cell membrane integrity of the microbes were quantitatively assessed. Correlation of membrane permeability and ATP levels with bacterial viability was statistically evaluated.</p> <p>Results</p> <p>PsA was relatively resistant to both H₂O₂(LD₅₀= 1.5 mM) and HOCl (LD₅₀= 0.035 mM). SA was susceptible to H₂O₂(LD₅₀= 0.1 mM) but resistant to HOCl (LD₅₀= 0.035 mM). Interestingly, KP was extremely resistant to high doses of H₂O₂(LD₅₀= 2.5-5.0 mM) but was very sensitive to low doses of HOCl (LD₅₀= 0.015 mM). BC was intermediate to resist both oxidants: H₂O₂(LD₅₀= 0.3-0.4 mM) and HOCl (LD₅₀= 0.025 mM). EC displayed the least resistance to H₂O₂(LD₅₀= 0.2-0.3 mM) and HOCl (LD₅₀= 0.015 mM). The identified profile of H₂O₂-resistance was KP > PsA > BC > EC > SA and the profile of HOCl-resistance PsA > SA > BC > EC > KP. Moreover, both oxidants affected ATP production and membrane integrity of the cells. However, the effects varied among the tested organisms and, the oxidant-mediated damage correlated differentially with the bacterial viability.</p> <p>Conclusions</p> <p>The order of HOCl-resistance identified herein best fits the clinical profile of CF infections. Even though oxidants are able to disrupt ATP production and cell membrane integrity, the degrees of damage vary among the organisms and correlate differentially with their viability.</p

Magnetostriction, elasticity, and D03 phase stability in Fe–Ga and Fe–Ga–Ge alloys

The contrast between the saturation tetragonal magnetostriction, λγ,2 = (3/2)λ100, of Fe1−xGax and Fe1−yGey, at compositions where both alloys exhibit D03 cubic symmetry (second peak region), was investigated. This region corresponds to x = 28 at. % Ga and y = 18 at. % Ge or, in terms of e/a = 2 x + 3 y + 1, to an e/a value of ∼1.55 for each of the alloys. Single crystal, slow-cooled, ternary Fe1−x−y GaxGey alloys with e/a ∼1.55 and gradually increasing y/x were investigated experimentally (magnetostriction, elasticity, powder XRD) and theoretically (density functional calculations). It was found that a small amount of Ge (y = 1.3) replacing Ga in the Fe–Ga alloy has a profound effect on the measured λγ,2. As y increases, the drop in λγ,2 is considerable, reaching negative values at y/x = 0.47. The two shear elastic constants c′ = (c11− c12)/2 and c44 measured for four compositions with 0.06 ≤ y/x ≤ 0.45 at 7 K range from 16 to 21 GPa and from 133 to 138 GPa, respectively. Large temperature dependence was observed for c′ but not for c44, a trend seen in other high-solute Fe alloys. The XRD analysis shows that the metastable D03 structure, observed previously in slow-cooled Fe–Ga at e/a = 1.55, is replaced with two phases, fcc L12 and hexagonal D019, at just 1.6 at. % Ge. The two are the stablephases of the assessed Fe–Ga phase diagram at x ∼ 28. Notably, at y = 7.8, only the D03phase (the equilibrium phase of Fe–Ge at e/a = 1.54) was found in the ternary alloy. The theory also shows that the D03 instability is removed for compositions with y ≥ 3.9, when D03 becomes the structure’s ground-state phase. Thus, the high, positive λγ,2 value for Fe–Ga at x = 28 could be the result of the high sensitivity of its metastable D03 structure

The secretome from bovine mammosphere-derived cells (MDC) promotes angiogenesis, epithelial cell migration, and contains factors associated with defense and immunity

Treatment of bovine mastitis with intramammary antibiotics is common, yet several concerns exist including failed efficacy for individual hosts or pathogens and the inability of approved drugs to revert mastitis-induced tissue damage to healthy tissue capable of returning to full milk production. These issues, in addition to aspects of public health such as accidental antibiotic residues in saleable milk and the potential for antimicrobial resistance, support the need to find alternative therapies for this costly disease. This study shows that the secretome, or collective factors, produced by mammosphere-derived cells (MDC) promotes angiogenesis, epithelial cell migration, and contains proteins associated with immunity and defense; all of which are necessary for healing damaged mammary gland tissue. Furthermore, we found that the MDC secretome remains effective after freezing and thawing, enhancing its therapeutic potential. Our results provide a foundation for further characterization of the individual secreted factors and the rationale for using the MDC secretome as a complementary treatment for bovine mastitis

Open Problems on Central Simple Algebras

We provide a survey of past research and a list of open problems regarding central simple algebras and the Brauer group over a field, intended both for experts and for beginners.Comment: v2 has some small revisions to the text. Some items are re-numbered, compared to v

Cost-minimization analysis in a blind randomized trial on small-incision versus laparoscopic cholecystectomy from a societal perspective: sick leave outweighs efforts in hospital savings

Background: After its introduction, laparoscopic cholecystectomy rapidly expanded around the world and was accepted the procedure of choice by consensus. However, analysis of evidence shows no difference regarding primary outcome measures between laparoscopic and small-incision cholecystectomy. In absence of clear clinical benefit it may be interesting to focus on the resource use associated with the available techniques, a secondary outcome measure. This study focuses on a difference in costs between laparoscopic and small-incision cholecystectomy from a societal perspective with emphasis on internal validity and generalisability Methods: A blinded randomized single-centre trial was conducted in a general teaching hospital in The Netherlands. Patients with reasonable to good health diagnosed with symptomatic cholecystolithiasis scheduled for cholecystectomy were included. Patients were randomized between laparoscopic and small-incision cholecystectomy. Total costs were analyzed from a societal perspective. Results: Operative costs were higher in the laparoscopic group using reusable laparoscopic instruments (difference 203 euro; 95% confidence interval 147 to 259 euro). There were no significant differences in the other direct cost categories (outpatient clinic and admittance related costs), indirect costs, and total costs. More than 60% of costs in employed patients were caused by sick leave. Conclusion: Based on differences in costs, small-incision cholecystectomy seems to be the preferred operative technique over the laparoscopic technique both from a hospital and societal cost perspective. Sick leave associated with convalescence after cholecystectomy in employed patients results in considerable costs to society

Zinc transporter gene expression is regulated by pro-inflammatory cytokines: a potential role for zinc transporters in beta-cell apoptosis?

<p>Abstract</p> <p>Background</p> <p>β-cells are extremely rich in zinc and zinc homeostasis is regulated by zinc transporter proteins. β-cells are sensitive to cytokines, interleukin-1β (IL-1β) has been associated with β-cell dysfunction and -death in both type 1 and type 2 diabetes. This study explores the regulation of zinc transporters following cytokine exposure.</p> <p>Methods</p> <p>The effects of cytokines IL-1β, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) on zinc transporter gene expression were measured in INS-1-cells and rat pancreatic islets. Being the more sensitive transporter, we further explored ZnT8 (Slc30A8): the effect of ZnT8 over expression on cytokine induced apoptosis was investigated as well as expression of the insulin gene and two apoptosis associated genes, BAX and BCL2.</p> <p>Results</p> <p>Our results showed a dynamic response of genes responsible for β-cell zinc homeostasis to cytokines: IL-1β down regulated a number of zinc-transporters, most strikingly ZnT8 in both islets and INS-1 cells. The effect was even more pronounced when mixing the cytokines. TNF-α had little effect on zinc transporter expression. IFN-γ down regulated a number of zinc transporters. Insulin expression was down regulated by all cytokines. ZnT8 over expressing cells were more sensitive to IL-1β induced apoptosis whereas no differences were observed with IFN-γ, TNF-α, or a mixture of cytokines.</p> <p>Conclusion</p> <p>The zinc transporting system in β-cells is influenced by the exposure to cytokines. Particularly ZnT8, which has been associated with the development of diabetes, seems to be cytokine sensitive.</p

HOXB13 is a susceptibility gene for prostate cancer: results from the International Consortium for Prostate Cancer Genetics (ICPCG)

Prostate cancer has a strong familial component but uncovering the molecular basis for inherited susceptibility for this disease has been challenging. Recently, a rare, recurrent mutation (G84E) in HOXB13 was reported to be associated with prostate cancer risk. Confirmation and characterization of this finding is necessary to potentially translate this information to the clinic. To examine this finding in a large international sample of prostate cancer families, we genotyped this mutation and 14 other SNPs in or flanking HOXB13 in 2,443 prostate cancer families recruited by the International Consortium for Prostate Cancer Genetics (ICPCG). At least one mutation carrier was found in 112 prostate cancer families (4.6%), all of European descent. Within carrier families, the G84E mutation was more common in men with a diagnosis of prostate cancer (194 of 382, 51%) than those without (42 of 137, 30%), P=9.9×10−8 [odds ratio 4.42 (95% confidence interval 2.56–7.64)]. A family-based association test found G84E to be significantly over-transmitted from parents to affected offspring (P=6.5×10−6). Analysis of markers flanking the G84E mutation indicates that it resides in the same haplotype in 95% of carriers, consistent with a founder effect. Clinical characteristics of cancers in mutation carriers included features of high-risk disease. These findings demonstrate that the HOXB13 G84E mutation is present in ~5% of prostate cancer families, predominantly of European descent, and confirm its association with prostate cancer risk. While future studies are needed to more fully define the clinical utility of this observation, this allele and others like it could form the basis for early, targeted screening of men at elevated risk for this common, clinically heterogeneous cancer.Electronic supplementary materialThe online version of this article (doi:10.1007/s00439-012-1229-4) contains supplementary material, which is available to authorized users