A New Class of Antibiotics Could Stop Drug-Resistant Bacteria in their Tracks

Bonnie Bassler, PhD is a world-renowned microbiologist and professor at Princeton University. She rose to fame, at least outside of the scientific community, after her 2009 TED talk entitled “How bacteria ‘talk’”. A stunning success, this 18-minute exposé into molecular biology is widely touted as one of the best general audience interpretations of research in the natural sciences. Bassler discusses the ways that bacteria communicate with one another, called quorum sensing, and explains its rather intricate mechanisms in clear and relatable terms. Quorum sensing relies upon small chemical messengers to transmit information between bacteria. The Bassler lab played a sizable role in dissecting these molecular transmissions and ‘rewriting’ them in an effort to devise modified antibiotics that block bacterial communication systems and shut down infections before bacterial populations grow out of control and cause damage to a human host. This piece in Synthesis attempts to break apart the scientific and medical ramifications of these cutting-edge, modernized antibiotics, while exposing the reader to the benefits of clear scientific communication. Bassler’s 2009 TED Talk has done more than simply catapult a scientist to fame – it has also encouraged political leaders to pay more attention to the growing antibiotic resistance in communities and expedite research in the field of microbiology for better bacterial-killing drugs

INFORMATION LITERACY STRATEGY: A CASE STUDY OF THE MANAGEMENT DEPARTMENT AT THE U.S. COAST GUARD ACADEMY

Izazov akademskog istraživanja nije pronaći informacije nego odabrati, organizirati i procijeniti valjanost i pouzdanost dostupnih informacija. Ovaj rad predstavlja strategiju Informacijske pismenosti (IP) koja je razvijena za unaprjeđenje IP vještina u preddiplomskom programu na Odjelu za menadžment USCG akademije. Autori demonstriraju IP strategije unutar kolegija kojeg predaju i pokazuju kako integrirana IP strategija odgovara na obrazovne IP standarde i zahtjeve. Rad navodi IP vježbe koje su razvijene i korištene u sljedećim kolegijima: Principi makroekonomije (prva godina); Organizacijsko ponašanje i vodstvo (druga godina); Uvod u biznis (druga godina); Marketing (treća godina); i Sustavi upravljanja informacijama (treća godina). Svaki kolegij pružen unutar studija Menadžmenta služi kao model razvoja IP vještina koje su strukturirane za razvoj IP vještina sekvencijalno od prve do zadnje godine preddiplomskog programa.The challenge of academic research is not to find information but to select, organize, and evaluate the validity and reliability of the information available. The paper presents the Information Literacy (IL) strategy that has been developed for the advancement of IL skills in an undergraduate program in the Management Department at the USCG Academy. The authors demonstrate IL strategies within the coursework they teach and how an integrated IL strategy responds to educational IL standards and requirements. The paper lists the IL exercises that were developed and used in the following courses: Macroeconomics Principles (freshman level); Organizational Behavior and Leadership (sophomore level); Introduction to Business (sophomore level); Marketing (junior level); and Management Information Systems (junior level). Each course provided within Management major serves as a model for the development of the IL skills that are structured to develop the IL skills sequentially from freshman to senior year in an undergraduate program

Strategija informacijske pismenosti: primjer Odjela za menadžment na Američkoj akademiji obalne straže

The challenge of academic research is not to find information but to select, organize, and evaluate the validity and reliability of the information available. The paper presents the Information Literacy (IL) strategy that has been developed for the advancement of IL skills in an undergraduate program in the Management Department at the USCG Academy. The authors demonstrate IL strategies within the coursework they teach and how an integrated IL strategy responds to educational IL standards and requirements. The paper lists the IL exercises that were developed and used in the following courses: Macroeconomics Principles (freshman level); Organizational Behavior and Leadership (sophomore level); Introduction to Business (sophomore level); Marketing (junior level); and Management Information Systems (junior level). Each course provided within Management major serves as a model for the development of the IL skills that are structured to develop the IL skills sequentially from freshman to senior year in an undergraduate program.Izazov akademskog istraživanja nije pronaći informacije nego odabrati, organizirati i procijeniti valjanost i pouzdanost dostupnih informacija. Ovaj rad predstavlja strategiju Informacijske pismenosti (IP) koja je razvijena za unaprjeđenje IP vještina u preddiplomskom programu na Odjelu za menadžment USCG akademije. Autori demonstriraju IP strategije unutar kolegija kojeg predaju i pokazuju kako integrirana IP strategija odgovara na obrazovne IP standarde i zahtjeve. Rad navodi IP vježbe koje su razvijene i korištene u sljedećim kolegijima: Principi makroekonomije (prva godina); Organizacijsko ponašanje i vodstvo (druga godina); Uvod u biznis (druga godina); Marketing (treća godina); i Sustavi upravljanja informacijama (treća godina). Svaki kolegij pružen unutar studija Menadžmenta služi kao model razvoja IP vještina koje su strukturirane za razvoj IP vještina sekvencijalno od prve do zadnje godine preddiplomskog programa

INFORMATION LITERACY STRATEGY: A CASE STUDY OF THE MANAGEMENT DEPARTMENT AT THE U.S. COAST GUARD ACADEMY

Izazov akademskog istraživanja nije pronaći informacije nego odabrati, organizirati i procijeniti valjanost i pouzdanost dostupnih informacija. Ovaj rad predstavlja strategiju Informacijske pismenosti (IP) koja je razvijena za unaprjeđenje IP vještina u preddiplomskom programu na Odjelu za menadžment USCG akademije. Autori demonstriraju IP strategije unutar kolegija kojeg predaju i pokazuju kako integrirana IP strategija odgovara na obrazovne IP standarde i zahtjeve. Rad navodi IP vježbe koje su razvijene i korištene u sljedećim kolegijima: Principi makroekonomije (prva godina); Organizacijsko ponašanje i vodstvo (druga godina); Uvod u biznis (druga godina); Marketing (treća godina); i Sustavi upravljanja informacijama (treća godina). Svaki kolegij pružen unutar studija Menadžmenta služi kao model razvoja IP vještina koje su strukturirane za razvoj IP vještina sekvencijalno od prve do zadnje godine preddiplomskog programa.The challenge of academic research is not to find information but to select, organize, and evaluate the validity and reliability of the information available. The paper presents the Information Literacy (IL) strategy that has been developed for the advancement of IL skills in an undergraduate program in the Management Department at the USCG Academy. The authors demonstrate IL strategies within the coursework they teach and how an integrated IL strategy responds to educational IL standards and requirements. The paper lists the IL exercises that were developed and used in the following courses: Macroeconomics Principles (freshman level); Organizational Behavior and Leadership (sophomore level); Introduction to Business (sophomore level); Marketing (junior level); and Management Information Systems (junior level). Each course provided within Management major serves as a model for the development of the IL skills that are structured to develop the IL skills sequentially from freshman to senior year in an undergraduate program

The Effect of DNA-Dependent Protein Kinase on Adeno-Associated Virus Replication

BACKGROUND: DNA-dependent protein kinase (DNA-PK) is a DNA repair enzyme and plays an important role in determining the molecular fate of the rAAV genome. However, the effect this cellular enzyme on rAAV DNA replication remains elusive. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we characterized the roles of DNA-PK on recombinant adeno-associated virus DNA replication. Inhibition of DNA-PK by a DNA-PK inhibitor or siRNA targeting DNA-PKcs significantly decreased replication of AAV in MO59K and 293 cells. Southern blot analysis showed that replicated rAAV DNA formed head-to-head or tail-to-tail junctions. The head-to-tail junction was low or undetectable suggesting AAV-ITR self-priming is the major mechanism for rAAV DNA replication. In an in vitro replication assay, anti-Ku80 antibody strongly inhibited rAAV replication, while anti-Ku70 antibody moderately decreased rAAV replication. Similarly, when Ku heterodimer (Ku70/80) was depleted, less replicated rAAV DNA were detected. Finally, we showed that AAV-ITRs directly interacted with Ku proteins. CONCLUSION/SIGNIFICANCE: Collectively, our results showed that that DNA-PK enhances rAAV replication through the interaction of Ku proteins and AAV-ITRs

Targeting Hepatitis B Virus with Zinc Finger Nucleases

Despite an existing effective vaccine, hepatitis B virus (HBV) remains a major public health concern. There are effective suppressive therapies for HBV, but they remain expensive and inaccessible to many, and not all patients respond well. Furthermore, HBV can persist as genomic covalently closed circular DNA (cccDNA) that remains in hepatocytes even during otherwise effective therapy and facilitates rebound in patients after treatment has stopped. Therefore, the need for an effective treatment that targets active and persistent HBV infections remains. As a novel approach to treat HBV, we have targeted the HBV genome for disruption to prevent viral reactivation and replication. We generated 3 zinc finger nucleases (ZFNs) that target sequences within the HBV polymerase, core and X genes. Upon the formation of ZFN-induced DNA double strand breaks (DSB), imprecise repair by non-homologous end joining leads to mutations that inactivate HBV genes. We delivered HBV-specific ZFNs using self-complementary adeno-associated virus (scAAV) vectors and tested their anti-HBV activity in HepAD38 cells. HBV-ZFNs efficiently disrupted HBV target sites by inducing site-specific mutations. Cytotoxicity was seen with one of the ZFNs. scAAV-mediated delivery of a ZFN targeting HBV polymerase resulted in complete inhibition of HBV DNA replication and production of infectious HBV virions in HepAD38 cells. This effect was sustained for at least 2 weeks following only a single treatment. Furthermore, high specificity was observed for all ZFNs, as negligible off-target cleavage was seen via high-throughput sequencing of 7 closely matched potential off-target sites. These results show that HBV-targeted ZFNs can efficiently inhibit active HBV replication and suppress the cellular template for HBV persistence, making them promising candidates for eradication therapy

Phenome-wide association analysis of LDL-cholesterol lowering genetic variants in PCSK9

Abstract: Background: We characterised the phenotypic consequence of genetic variation at the PCSK9 locus and compared findings with recent trials of pharmacological inhibitors of PCSK9. Methods: Published and individual participant level data (300,000+ participants) were combined to construct a weighted PCSK9 gene-centric score (GS). Seventeen randomized placebo controlled PCSK9 inhibitor trials were included, providing data on 79,578 participants. Results were scaled to a one mmol/L lower LDL-C concentration. Results: The PCSK9 GS (comprising 4 SNPs) associations with plasma lipid and apolipoprotein levels were consistent in direction with treatment effects. The GS odds ratio (OR) for myocardial infarction (MI) was 0.53 (95% CI 0.42; 0.68), compared to a PCSK9 inhibitor effect of 0.90 (95% CI 0.86; 0.93). For ischemic stroke ORs were 0.84 (95% CI 0.57; 1.22) for the GS, compared to 0.85 (95% CI 0.78; 0.93) in the drug trials. ORs with type 2 diabetes mellitus (T2DM) were 1.29 (95% CI 1.11; 1.50) for the GS, as compared to 1.00 (95% CI 0.96; 1.04) for incident T2DM in PCSK9 inhibitor trials. No genetic associations were observed for cancer, heart failure, atrial fibrillation, chronic obstructive pulmonary disease, or Alzheimer’s disease – outcomes for which large-scale trial data were unavailable. Conclusions: Genetic variation at the PCSK9 locus recapitulates the effects of therapeutic inhibition of PCSK9 on major blood lipid fractions and MI. While indicating an increased risk of T2DM, no other possible safety concerns were shown; although precision was moderate

Genetic Barcodes Enable Quantitative Mapping of Operator Mutants to Gene Expression

Every organism has intricate regulatory networks that enable them to sense, move, and interact with complex environments. In E. coli, transcription factors (TFs) bind to short sequences upstream of genes, called operators, to repress or activate gene expression. Our lab has previously derived and validated a biophysical model of transcription regulation -the repression of a gene by LacI - demonstrating that repressorcopy number, the energy of TF:operator binding, and genome size all play crucial roles in determining the quantitative features of gene expression. But LacI is a small fish in the cellular pond; we must develop experimental methods that enable us to map how any TF:operator pair imparts predictable gene expression, and do so in a high-throughput manner without sacrificing quantitation. We report a high-throughput method to randomly mutagenize large libraries of operator sequences and quantify their resulting gene expression. Briefly, mutagenized binding sites for a TF are “mapped” to a random DNA barcode using next-generation sequencing. The native gene regulated by that TF is then inserted between the mutated operators and barcodes, and these libraries are genomically-integrated into E. coli. The relative abundance of cells carrying each operator mutant can be determined via DNA-sequencing of the barcodes, while their gene expression can be measured by quantitative RNA-sequencing of barcodes using a mixture of Unique Molecular Identifiers and other methods to minimize bias. These data will inform mathematical models for the de novo prediction of gene expression from regulatory sequences