Comparison of the in vitro and in vivo susceptibilities of Burkholderia mallei to Ceftazidime and Levofloxacin

<p>Abstract</p> <p>Background</p> <p><it>Burkholderia mallei </it>is a zoonotic Gram negative bacterium which primarily infects solipeds but can cause lethal disease in humans if left untreated. The effect of two antibiotics with different modes of action on <it>Burkholderia mallei </it>strain ATCC23344 was investigated by using <it>in vitro </it>and <it>in vivo </it>studies.</p> <p>Results</p> <p>Determination of minimal inhibitory concentrations (MICs) <it>in vitro </it>was done by the agar diffusion method and the dilution method. The MICs of levofloxacin and ceftazidime were in the similar range, 2.5 and 5.0 μg/ml, respectively. Intracellular susceptibility of the bacterium to these two antibiotics in J774A.1 mouse macrophages <it>in vitro </it>was also investigated. Macrophages treated with antibiotics demonstrated uptake of the drugs and reduced bacterial loads <it>in vitro</it>. The efficacy of ceftazidime and levofloxacin were studied in BALB/c mice as post-exposure treatment following intranasal <it>B. mallei </it>infection. Intranasal infection with 5 × 10⁵CFUs of <it>B. mallei </it>resulted in 90% death in non-treated control mice. Antibiotic treatments 10 days post-infection proved to be effective <it>in vivo </it>with all antibiotic treated mice surviving to day 34 post-infection. The antibiotics did not result in complete clearance of the bacterial infection and presence of the bacteria was found in lungs and spleens of the survivors, although bacterial burden recovered from levofloxacin treated animals appeared reduced compared to ceftazidime.</p> <p>Conclusion</p> <p>Both antibiotics demonstrated utility for the treatment of glanders, including the ability for intracellular penetration and clearance of organisms <it>in vitro</it>.</p

Host immunity in the protective response to vaccination with heat-killed Burkholderia mallei

<p>Abstract</p> <p>Background</p> <p>We performed initial cell, cytokine and complement depletion studies to investigate the possible role of these effectors in response to vaccination with heat-killed <it>Burkholderia mallei </it>in a susceptible BALB/c mouse model of infection.</p> <p>Results</p> <p>While protection with heat-killed bacilli did not result in sterilizing immunity, limited protection was afforded against an otherwise lethal infection and provided insight into potential host protective mechanisms. Our results demonstrated that mice depleted of either B cells, TNF-α or IFN-γ exhibited decreased survival rates, indicating a role for these effectors in obtaining partial protection from a lethal challenge by the intraperitoneal route. Additionally, complement depletion had no effect on immunoglobulin production when compared to non-complement depleted controls infected intranasally.</p> <p>Conclusion</p> <p>The data provide a basis for future studies of protection via vaccination using either subunit or whole-organism vaccine preparations from lethal infection in the experimental BALB/c mouse model. The results of this study demonstrate participation of B220⁺cells and pro-inflammatory cytokines IFN-γ and TNF-α in protection following HK vaccination.</p

In vivo Bioluminescence Imaging of Burkholderia mallei Respiratory Infection and Treatment in the Mouse Model

Bioluminescent imaging (BLI) technology is a powerful tool for monitoring infectious disease progression and treatment approaches. BLI is particularly useful for tracking fastidious intracellular pathogens that might be difficult to recover from certain organs. Burkholderia mallei, the causative agent of glanders, is a facultative intracellular pathogen and has been classified by the CDC as a Category B select agent due to its highly infectious nature and potential use as a biological weapon. Very little is known regarding pathogenesis or treatment of glanders. We investigated the use of bioluminescent reporter constructs to monitor the dynamics of infection as well as the efficacy of therapeutics for B. mallei in real-time. A stable luminescent reporter B. mallei strain was created using the pUTmini-Tn5::luxKm2 plasmid and used to monitor glanders in the BALB/c murine model. Mice were infected via the intranasal route with 5 × 103 bacteria and monitored by BLI at 24, 48, and 72 h. We verified that our reporter construct maintained similar virulence and growth kinetics compared to wild-type B. mallei and confirmed that it maintains luminescent stability in the presence or absence of antibiotic selection. The luminescent signal was initially seen in the lungs, and progressed to the liver and spleen over the course of infection. We demonstrated that antibiotic treatment 24 h post-infection resulted in reduction of bioluminescence that can be attributed to decreased bacterial burden in target organs. These findings suggest that BLI can be used to monitor disease progression and efficacy of therapeutics during glanders infections. Finally, we report an alternative method to mini-Tn5::luxKm2 transposon using mini-Tn7-lux elements that insert site-specifically at known genomic attachment sites and that can also be used to tag bacteria

Large Effects from Small Exposures. I. Mechanisms for Endocrine-Disrupting Chemicals with Estrogenic Activity

Reproduced with permission from Environmental Health Perspectives. doi:10.1289/ehp.5494Information concerning the fundamental mechanisms of action of both natural and environmental hormones, combined with information concerning endogenous hormone concentrations, reveals how endocrine-disrupting chemicals with estrogenic activity (EEDCs) can be active at concentrations far below those currently being tested in toxicological studies. Using only very high doses in toxicological studies of EEDCs thus can dramatically underestimate bioactivity. Specifically: a) The hormonal action mechanisms and the physiology of delivery of EEDCs predict with accuracy the low-dose ranges of biological activity, which have been missed by traditional toxicological testing. b) Toxicology assumes that it is valid to extrapolate linearly from high doses over a very wide dose range to predict responses at doses within the physiological range of receptor occupancy for an EEDC; however, because receptor-mediated responses saturate, this assumption is invalid. c) Furthermore, receptor-mediated responses can first increase and then decrease as dose increases, contradicting the assumption that dose-response relationships are monotonic. d) Exogenous estrogens modulate a system that is physiologically active and thus is already above threshold, contradicting the traditional toxicological assumption of thresholds for endocrine responses to EEDCs. These four fundamental issues are problematic for risk assessment methods used by regulatory agencies, because they challenge the traditional use of extrapolation from high-dose testing to predict responses at the much lower environmentally relevant doses.Support during the preparation of this manuscript was provided by the W. Alton Jones Foundation to K.A.T, as well as by grants from the National Institutes of Health (NIH) (CA50354) and the University of Missouri (VMFC0018) to W.V.W and NIH (ES08293 and ES11283), U.S. Environmental Protection Agency (U914991), and University of Missouri Research Board to F.v.S

Polysaccharide Specific Monoclonal Antibodies Provide Passive Protection against Intranasal Challenge with Burkholderia pseudomallei

Burkholderia pseudomallei is a Gram-negative bacillus that is the causative agent of melioidosis. The bacterium is inherently resistant to many antibiotics and mortality rates remain high in endemic areas. The lipopolysaccharide (LPS) and capsular polysaccharide (CPS) are two surface-associated antigens that contribute to pathogenesis. We previously developed two monoclonal antibodies (mAbs) specific to the CPS and LPS; the CPS mAb was shown to identify antigen in serum and urine from melioidosis patients. The goal of this study was to determine if passive immunization with CPS and LPS mAbs alone and in combination would protect mice from a lethal challenge with B. pseudomallei. Intranasal (i.n.) challenge experiments were performed with B. pseudomallei strains 1026b and K96423. Both mAbs provided significant protection when administered alone. A combination of mAbs was protective when low doses were administered. In addition, combination therapy provided a significant reduction in spleen colony forming units (cfu) compared to results when either the CPS or LPS mAbs were administered alone

Core competencies for pain management: results of an interprofessional consensus summit.

ObjectiveThe objective of this project was to develop core competencies in pain assessment and management for prelicensure health professional education. Such core pain competencies common to all prelicensure health professionals have not been previously reported.MethodsAn interprofessional executive committee led a consensus-building process to develop the core competencies. An in-depth literature review was conducted followed by engagement of an interprofessional Competency Advisory Committee to critique competencies through an iterative process. A 2-day summit was held so that consensus could be reached.ResultsThe consensus-derived competencies were categorized within four domains: multidimensional nature of pain, pain assessment and measurement, management of pain, and context of pain management. These domains address the fundamental concepts and complexity of pain; how pain is observed and assessed; collaborative approaches to treatment options; and application of competencies across the life span in the context of various settings, populations, and care team models. A set of values and guiding principles are embedded within each domain.ConclusionsThese competencies can serve as a foundation for developing, defining, and revising curricula and as a resource for the creation of learning activities across health professions designed to advance care that effectively responds to pain

Caby Photometry of the Hyades: Comparisons to the Field Stars

Intermediate-band photometry of the Hyades cluster on the Caby system is presented for dwarf stars ranging from spectral type A through late K. A mean hk, b-y relation is constructed using only single stars without anomalous atmospheres and compared to the field stars of the solar neighborhood. For the F dwarfs, the Hyades relation defines an approximate LOWER bound in the two-color diagram, consistent with an [Fe/H] between +0.10 and +0.15. These index-color diagrams follow the common convention of presenting stars with highest abundance at the bottom of the plot although the index values for the metal-rich stars are numerically larger. For field F dwarfs in the range [Fe/H] between +0.4 and -1.0, [Fe/H] = -5.6 delta-hk + 0.125, with no evidence for a color dependence in the slope. For the G and K dwarfs, the Hyades mean relation crosses the field star distribution in the two-color diagram, defining an approximate UPPER bound for the local disk stars. Stars found above the Hyades stars fall in at least one of three categories: [Fe/H] below -0.7, [Fe/H] above that of the Hyades, or chromospherically active. It is concluded that, contrary to the predictions of model atmospheres, the hk index for cool dwarfs at a given color hits a maximum value for stars below solar composition and, with increasing [Fe/H] above some critical value, declines. This trend is consistent, however, with the predictions from synthetic indices based upon much narrower Ca filters where the crossover is caused by the metallicity sensitivity of b-y.Comment: 13 pages, 9 eps figures, 1 tex table, 1 ascii tabl

Bisphenol A Is Released from Used Polycarbonate Animal Cages into Water at Room Temperature

doi:10.1289/ehp.5993Bisphenol A (BPA) is a monomer with estrogenic activity that is used in the production of food packaging, dental sealants, polycarbonate plastic, and many other products. The monomer has previously been reported to hydrolyze and leach from these products under high heat and alkaline conditions, and the amount of leaching increases as a function of use. We examined whether new and used polycarbonate animal cages passively release bioactive levels of BPA into water at room temperature and neutral pH. Purified water was incubated at room temperature in new polycarbonate and polysulfone cages and used (discolored) polycarbonate cages, as well as control (glass and used polypropylene) containers. The resulting water samples were characterized with gas chromatography/mass spectrometry (GC/MS) and tested for estrogenic activity using an MCF-7 human breast cancer cell proliferation assay. Significant estrogenic activity, identifiable as BPA by GC/MS (up to 310 µg/L), was released from used polycarbonate animal cages. Detectable levels of BPA were released from new polycarbonate cages (up to 0.3 µg/L) as well as new polysulfone cages (1.5 µg/L), whereas no BPA was detected in water incubated in glass and used polypropylene cages. Finally, BPA exposure as a result of being housed in used polycarbonate cages produced a 16% increase in uterine weight in prepubertal female mice relative to females housed in used polypropylene cages, although the difference was not statistically significant. Our findings suggest that laboratory animals maintained in polycarbonate and polysulfone cages are exposed to BPA via leaching, with exposure reaching the highest levels in old cages.Support during the preparation of this manuscript was provided by grants from the National Institutes of Health (CA50354) and the University of Missouri (VMFC0018) to W.V.W., NIH (ES08293 and ES11283) to F.v.S., and the U.S.G.S