A practical laboratory method to determine ceftazidime-avibactam-aztreonam synergy in patients with New Delhi Metallo-beta-lactamase (NDM) producing Enterobacterales infection

Background: In response to infection with New Delhi Metallo-beta-lactamase (NDM) producing Enterobacterales, combination antimicrobial therapy with ceftazidime/avibactam (CAZ/AVI) plus aztreonam (ATM) has been explored. This study evaluated a practical laboratory method of testing for clinically significant synergy between CAZ/AVI+ATM in NDM producing Enterobacterales. Methods: Minimum inhibitory concentration (MIC) of clinical NDM producing isolates were determined for ATM alone and CAZ/AVI+ATM using broth dilution. Restoration of ATM breakpoint following the addition of CAZ/AVI was explored. A CAZ/AVI E-test/ATM disc method was compared to broth dilution. Results: Of 43 isolates, 33/43 (77%) isolates were ATM resistant (median [range] MIC=56 [16 – 512] mg/L). Addition of CAZ/AVI restored the ATM breakpoint (MIC <4mg/L) in 29/33 (89%) of resistant isolates. Overall, the E-test/disc method correlated with findings from broth dilution in 35/43 (81%) of cases. E-test/disc sensitivity was 77% and specificity 85%. Positive predictive value was 92% and negative predictive value 61%. Conclusion: CAZ/AVI+ATM demonstrated significant synergy in most ATM resistant NDM producing Enterobacterales. The E-test/disc method is a quick, reproducible, and reliable method of testing for clinically relevant synergy in the microbiology laboratory

Long-Term Systemic Myostatin Inhibition via Liver-Targeted Gene Transfer in Golden Retriever Muscular Dystrophy

Duchenne muscular dystrophy (DMD) is a lethal, X-linked recessive disease affecting 1 in 3,500 newborn boys for which there is no effective treatment or cure. One novel strategy that has therapeutic potential for DMD is inhibition of myostatin, a negative regulator of skeletal muscle mass that may also promote fibrosis. Therefore, our goal in this study was to evaluate systemic myostatin inhibition in the golden retriever model of DMD (GRMD). GRMD canines underwent liver-directed gene transfer of a self-complementary adeno-associated virus type 8 vector designed to express a secreted dominant-negative myostatin peptide (n =4) and were compared with age-matched, untreated GRMD controls (n =3). Dogs were followed with serial magnetic resonance imaging (MRI) for 13 months to assess cross-sectional area and volume of skeletal muscle, then euthanized so that tissue could be harvested for morphological and histological analysis. We found that systemic myostatin inhibition resulted in increased muscle mass in GRMD dogs as assessed by MRI and confirmed at tissue harvest. We also found that hypertrophy of type IIA fibers was largely responsible for the increased muscle mass and that reductions in serum creatine kinase and muscle fibrosis were associated with long-term myostatin inhibition in GRMD. This is the first report describing the effects of long-term, systemic myostatin inhibition in a large-animal model of DMD, and we believe that the simple and effective nature of our liver-directed gene-transfer strategy makes it an ideal candidate for evaluation as a novel therapeutic approach for DMD patients

Long-term Restoration of Cardiac Dystrophin Expression in Golden Retriever Muscular Dystrophy Following rAAV6-mediated Exon Skipping

Although restoration of dystrophin expression via exon skipping in both cardiac and skeletal muscle has been successfully demonstrated in the mdx mouse, restoration of cardiac dystrophin expression in large animal models of Duchenne muscular dystrophy (DMD) has proven to be a challenge. In large animals, investigators have focused on using intravenous injection of antisense oligonucleotides (AO) to mediate exon skipping. In this study, we sought to optimize restoration of cardiac dystrophin expression in the golden retriever muscular dystrophy (GRMD) model using percutaneous transendocardial delivery of recombinant AAV6 (rAAV6) to deliver a modified U7 small nuclear RNA (snRNA) carrying antisense sequence to target the exon splicing enhancers of exons 6 and 8 and correct the disrupted reading frame. We demonstrate restoration of cardiac dystrophin expression at 13 months confirmed by reverse transcription-PCR (RT-PCR) and immunoblot as well as membrane localization by immunohistochemistry. This was accompanied by improved cardiac function as assessed by cardiac magnetic resonance imaging (MRI). Percutaneous transendocardial delivery of rAAV6 expressing a modified U7 exon skipping construct is a safe, effective method for restoration of dystrophin expression and improvement of cardiac function in the GRMD canine and may be easily translatable to human DMD patients

Microorganisms in metalworking fluids: Current issues in research and management

The microbial contamination of water miscible metalworking fl uids (MWFs) is a serious problem in metal industry. A good maintenance of MWF re-circulation systems can extend the lifetime of coolants and ensure the quality of the tools produced. In MWFs, as in the other water-based environments, microorganisms usually live in the form of biofi lms, the communities of bacteria and fungi attached to the surface of sumps, metal parts and also to each other. Biofi lms exhibit very high resistance to biocides. The effect of biocides that are used as additives to MWFs to control the growth of the bacterial and fungal microbiomes (microorganisms characteristic to the individual coolant system) have become the subject of research only in recent years. There are also only sparse reports on the impact of biocides on microorganisms growing in biofi lms in MWF installations. Fast growing mycobacteria are important members of these biofi lm communities. Their presence has recently been linked with the occurrence of cases of hypersensitivity pneumonitis, a serious respiratory disorder in the metal industry employees. The new, relatively fast and inexpensive techniques to assess the species diversity within MWF microbiomes and their population size should be developed in order to control the microorganisms’ proliferation in MWFs and to diminish the occupational exposure to harmful bioaerosols in metal industry

An enlightened sense of virtue: Morals and manners in eighteenth-century American periodical literature

The purpose of this study was to determine the extent to which the Enlightenment reached middle- and lower-class colonial Americans prior to independence and revolution. Essays on morals and manners appearing in five periodicals published along the southern and mid-Atlantic coast between the years 1750 and 1775 were analyzed for markers of Enlightenment thought. These were isolated from a review of well-known social and intellectual histories of the period. An advertiser list and a subscription list provided documented readers for two of the five periodicals, which were then analyzed for income, occupation, and socio-economic status using demographic studies as well as an early taxables list of colonial Philadelphians. Enlightened ideas of rational criticism, tolerant civility, a belief in progress, and a simplified moral philosophy based on the practice of modest virtues were found to have framed the periodical essays. The essays covered a variety of issues that touched the social, personal, and at times, very intimate lives of colonial Americans. The colonial readers of these essays emerged from across the socio-economic spectrum, with a substantial number coming from the middle and working classes. The periodical essays, as a whole, provided these colonists with an enlightened moral guide to living a good and virtuous life, and the effects of such a guide were to imbue its readers with a sense of empowerment and to contribute to the evolution of American republicanism

Secretion of Proteases by Pseudomonas aeruginosa Biofilms Exposed to Ciprofloxacin

Pseudomonas aeruginosa proteases are considered important virulence factors which damage host tissues and interfere with host antibacterial defense mechanisms. P. aeruginosa biofilm cells are not completely killed by antibacterials, and therefore this study addresses the question whether ciprofloxacin attenuates the virulence of biofilm communities by abolishing their secretion of proteases. The surviving cells of the colony biofilms studied, despite their cyclical exposure to four doses of ciprofloxacin at bactericidal concentrations (one dose a day), still secreted active proteases to the environment surrounding the biofilms. The biofilm cells secreted elastase B (LasB) over the duration of the experiments as confirmed by Western immunoblot analysis. The colony biofilms did not secrete LasA—a protease with staphylolytic activity. The same profiles on zymogram gels with gelatin were observed for the proteases secreted by both ciprofloxacin-exposed and unexposed (control) biofilms. Total proteolytic activities of the colony biofilms studied were significantly reduced after exposure to ciprofloxacin at bactericidal concentrations—after 96 h of exposure they dropped to 38% for the strain intermediate resistant to ciprofloxacin and to 65% for the strain highly resistant to the antibiotic, relative to the control biofilms. The surviving cells of the colony biofilms after their release into a fresh medium displayed transient increased resistance to ciprofloxacin compared to their planktonic counterparts