The Antibacterial Drug Candidate SBC3 is a Potent Inhibitor of Bacterial Thioredoxin Reductase

Antibiotic resistance is a growing problem for public health and associated with increasing economic costs and mortality rates. Silver and silver-related compounds have been used for centuries due to their antimicrobial properties. In this work, we show that 1,3-dibenzyl-4,5-diphenyl-imidazol-2-ylidene silver(I) acetate/NHC*-Ag-OAc (SBC3) is a reversible, high affinity inhibitor of E. coli thioredoxin reductase (TrxR; K_i=10.8±1.2 nM). Minimal inhibition concentration (MIC) tests with different E. coli and P. aeruginosa strains demonstrated that SBC3 can efficiently inhibit bacterial cell growth, especially in combination with established antibiotics like gentamicin. Our results show that SBC3 is a promising antibiotic drug candidate targeting bacterial TrxR.     

Congenital Diseases of DNA Replication: Clinical Phenotypes and Molecular Mechanisms

Deoxyribonucleic acid (DNA) replication can be divided into three major steps: initiation, elongation and termination. Each time a human cell divides, these steps must be reiteratively carried out. Disruption of DNA replication can lead to genomic instability, with the accumulation of point mutations or larger chromosomal anomalies such as rearrangements. While cancer is the most common class of disease associated with genomic instability, several congenital diseases with dysfunctional DNA replication give rise to similar DNA alterations. In this review, we discuss all congenital diseases that arise from pathogenic variants in essential replication genes across the spectrum of aberrant replisome assembly, origin activation and DNA synthesis. For each of these conditions, we describe their clinical phenotypes as well as molecular studies aimed at determining the functional mechanisms of disease, including the assessment of genomic stability. By comparing and contrasting these diseases, we hope to illuminate how the disruption of DNA replication at distinct steps affects human health in a surprisingly cell-type-specific manner.     

Allele-Specific Inhibition of Histone Demethylases

Histone demethylases play a critical role in mammalian gene expression by removing methyl groups from lysine residues in degree- and site-specific manner. To specifically interrogate members and isoforms of this class of enzymes, we have developed demethylase variants with an expanded active site. The mutant enzymes are capable of performing lysine demethylation with wild-type proficiency, but are sensitive to inhibition by cofactor-competitive molecules embellished with a complementary steric “bump”. The selected inhibitors show more than 20-fold selectivity over the wild-type demethylase, thus overcoming issues typical to pharmacological and genetic approaches. The mutant–inhibitor pairs are shown to act on a physiologically relevant full-length substrate. By engineering a conserved amino acid to achieve member-specific perturbation, this study provides a general approach for studying histone demethylases in diverse cellular processes.     

Electrochemical Characterization of Isolated Nitrogenase Cofactors from Azotobacter vinelandii

The nitrogenase cofactors are structurally and functionally unique in biological chemistry. Despite a substantial amount of spectroscopic characterization of protein-bound and isolated nitrogenase cofactors, electrochemical characterization of these cofactors and their related species is far from complete. Herein we present voltammetric studies of three isolated nitrogenase cofactor species: the iron–molybdenum cofactor (M-cluster), iron–vanadium cofactor (V-cluster), and a homologue to the iron–iron cofactor (L-cluster). We observe two reductive events in the redox profiles of all three cofactors. Of the three, the V-cluster is the most reducing. The reduction potentials of the isolated cofactors are significantly more negative than previously measured values within the molybdenum–iron and vanadium–iron proteins. The outcome of this study provides insight into the importance of the heterometal identity, the overall ligation of the cluster, and the impact of the protein scaffolds on the overall electronic structures of the cofactors.     

Challenges in forming inferences from limited data: a case study of malaria parasite maturation

Inferring biological processes from population dynamics is a common challenge in ecology, particularly when faced with incomplete data. This challenge extends to inferring parasite traits from within-host infection dynamics. We focus on rodent malaria infections (Plasmodium berghei), a system for which previous work inferred an immune-mediated extension in the length of the parasite development cycle within red blood cells. By developing a system of delay-differential equations to describe within-host infection dynamics and simulating data, we demonstrate the potential to obtain biased estimates of parasite (and host) traits when key biological processes are not considered. Despite generating infection dynamics using a fixed parasite developmental cycle length, we find that known sources of measurement bias in parasite stage and abundance data can affect estimates of parasite developmental duration, with stage misclassification driving inferences about extended cycle length. We discuss alternative protocols and statistical methods that can mitigate such misestimation.     

Virtual fencing technology to intensively graze lactating dairy cattle. II: Effects on cow welfare and behavior

Virtual fencing technology uses a neckband-mounted device to deliver an audio cue when an animal nears a virtual boundary that is set via a global positioning system, followed by an electrical stimulus if it crosses the boundary. The flexibility offered by this technology could revolutionize grazing management on dairy farms, but its application and effects on lactating dairy cattle have not been assessed. This experiment reports on the effects of an electric or a virtual front-fence on dairy cow behavior and welfare. Two temporally separated treatments were applied to a herd of 30 multiparous cows. Cows were provided an estimated 14 to 15 kg of dry matter/cow of fresh pasture in a new paddock every 24 h. From d 1 to 10 cows were grazed using a conventional electric front-fence (control treatment) and from d 14 to 23 they were grazed using a virtual front-fence (eShepherd, Agersens Pty Ltd.). Cows were trained to the technology from d 11 to 13. The milk production and live weight of individual cows were recorded daily. Cortisol concentrations were obtained from milk samples collected from individual cows on 3 d during each of the control and the virtual fence grazing periods, plus the first day of training. From d 6 of the experiment, 6 focal cows were fitted with a RumiWatch (Itin + Hoch GmbH) noseband sensor to monitor grazing and ruminating time, and 8 focal cows were fitted with an IceTag (IceRobotics Ltd.) sensors to monitor activity. Milk production, live weight, and the time cows spent standing and lying did not differ between the electric and virtual fence periods. Milk cortisol concentrations, activity, and the times spent ruminating and grazing were comparable between the electric and early virtual fence periods (i.e., d 1–3 with a virtual fence). However, at d 4 to 6 with a virtual fence, activity (steps taken and motion index) and time spent grazing were lower, and time spent ruminating was greater, compared with an electric fence. Further, least significant difference tests suggest milk cortisol concentrations were higher at d 5 with a virtual fence than at d 8 with an electric fence and d 1 with a virtual fence. We conclude there is no evidence of behavioral and welfare effects of virtual fencing on dairy cows in the days immediately following implementation of the technology in a simple intensive grazing regimen, but a longer study is required to fully elucidate effects beyond this period.     

Biofilm‐Eradicating Properties of Quaternary Ammonium Amphiphiles: Simple Mimics of Antimicrobial Peptides

Bacterial biofilms are difficult to eradicate because of reduced antibiotic sensitivity and altered metabolic processes; thus, the development of new approaches to biofilm eradication is urgently needed. Antimicrobial peptides (AMPs) and quaternary ammonium cations (QACs) are distinct, yet well‐known, classes of antibacterial compounds. By mapping the general regions of charge and hydrophobicity of QACs onto AMP structures, we designed a small library of QACs to serve as simple AMP mimics. In order to explore the role that cationic charge plays in biofilm eradication, structures were varied with respect to cationic character, distribution of charge, and alkyl side chain. The reported compounds possess minimum biofilm eradication concentrations (MBEC) as low as 25 μM against Gram‐positive biofilms, making them the most active anti‐biofilm structures reported to date. These potent AMP mimics were synthesized in 1–2 steps and hint at the minimal structural requirements for biofilm destruction.