Total synthesis of barettin: Model study of specialized aldol condensation to directly access diketopiperazine targets

We synthesized (±)-(Z)-barettin, a potent anti-biofouler, with 31 % yield as a mono-TFA salt over seven steps with only one chromatographic separation. The route is efficient, proceeds under mild conditions, and is optimized to reduce formation of undesired byproducts otherwise seen in the aldol condensations and the Boc protection. Our route enables us to 1) utilize a DKP condensation with an unstable aliphatic aldehyde, 2) showcase how the DKP condensation provides direct access to a bioactive scaffold, and 3) suggest a simple, high yielding route to barettin and analogues consisting of only one chromatographic separation

Methanol Accelerates DMPC Flip-Flop and Transfer: A SANS Study on Lipid Dynamics

© 2019 Biophysical Society Methanol is a common solubilizing agent used to study transmembrane proteins/peptides in biological and synthetic membranes. Using small angle neutron scattering and a strategic contrast-matching scheme, we show that methanol has a major impact on lipid dynamics. Under increasing methanol concentrations, isotopically distinct 1,2-dimyristoyl-sn-glycero-3-phosphocholine large unilamellar vesicle populations exhibit increased mixing. Specifically, 1,2-dimyristoyl-sn-glycero-3-phosphocholine transfer and flip-flop kinetics display linear and exponential rate enhancements, respectively. Ultimately, methanol is capable of influencing the structure-function relationship associated with bilayer composition (e.g., lipid asymmetry). The use of methanol as a carrier solvent, despite better simulating some biological conditions (e.g., antimicrobial attack), can help misconstrue lipid scrambling as the action of proteins or peptides, when in actuality it is a combination of solvent and biological agent. As bilayer compositional stability is crucial to cell survival and protein reconstitution, these results highlight the importance of methanol, and solvents in general, in biomembrane and proteolipid studies

Time-resolved SANS reveals pore-forming peptides cause rapid lipid reorganization

Cells depend on proper lipid transport and their precise distribution for vital cellular function. Disruption of such lipid organization can be initiated by external agents to cause cell death. Here, we investigate two antimicrobial pore-forming peptides, alamethicin and melittin, and their influence on lipid intervesicular exchange and transverse lipid diffusion (i.e. flip-flop) in model lipid vesicles. Small angle neutron scattering (SANS) and a strategic contrast matching scheme show the mixing of two isotopically distinct dimyristoylphosphocholine (DMPC) vesicle populations is promoted upon the addition of high (1/40) and low (1/150, 1/1000) peptide-to-lipid (P/L) molar ratios. Parsing out the individual exchange and flip-flop rate constants revealed that alamethicin increases both DMPC flip-flop and exchange by ≈2-fold when compared to methanol alone (the carrier solvent of the peptides). On the other hand, melittin affected DMPC flip-flop by a factor of 1 to 4 depending on the concentration, but had little effect on inter-vesicle lipid exchange at low P/L ratios. Thermodynamic parameters measured at high protein concentrations (P/L = 1/40) yielded remarkable similarity in the values obtained for both peptides, indicating likeness in their mechanism of action on lipid motion despite differences in their proposed oligomeric pore structures. The entropic contributions to the free energy of activation became favorable upon peptide addition, while the enthalpy of activation remained the major barrier to lipid exchange and flip-flop. This journal i

Transverse lipid organization dictates bending fluctuations in model plasma membranes

© 2020 The Royal Society of Chemistry. Membrane undulations play a vital role in many biological processes, including the regulation of membrane protein activity. The asymmetric lipid composition of most biological membranes complicates theoretical description of these bending fluctuations, yet experimental data that would inform any such a theory is scarce. Here, we used neutron spin-echo (NSE) spectroscopy to measure the bending fluctuations of large unilamellar vesicles (LUV) having an asymmetric transbilayer distribution of high- and low-melting lipids. The asymmetric vesicles were prepared using cyclodextrin-mediated lipid exchange, and were composed of an outer leaflet enriched in egg sphingomyelin (ESM) and an inner leaflet enriched in 1-palmitoyl-2-oleoyl-phosphoethanolamine (POPE), which have main transition temperatures of 37 °C and 25 °C, respectively. The overall membrane bending rigidity was measured at three temperatures: 15 °C, where both lipids are in a gel state; 45 °C, where both lipids are in a fluid state; and 30 °C, where there is gel-fluid co-existence. Remarkably, the dynamics for the fluid asymmetric LUVs (aLUVs) at 30 °C and 45 °C do not follow trends predicted by their symmetric counterparts. At 30 °C, compositional asymmetry suppressed the bending fluctuations, with the asymmetric bilayer exhibiting a larger bending modulus than that of symmetric bilayers corresponding to either the outer or inner leaflet. We conclude that the compositional asymmetry and leaflet coupling influence the internal dissipation within the bilayer and result in membrane properties that cannot be directly predicted from corresponding symmetric bilayers

The antioxidant vitamin E as a membrane raft modulator: Tocopherols do not abolish lipid domains

© 2020 Elsevier B.V. The antioxidant vitamin E is a commonly used vitamin supplement. Although the multi-billion dollar vitamin and nutritional supplement industry encourages the use of vitamin E, there is very little evidence supporting its actual health benefits. Moreover, vitamin E is now marketed as a lipid raft destabilizing anti-cancer agent, in addition to its antioxidant behaviour. Here, we studied the influence of vitamin E and some of its vitamers on membrane raft stability using phase separating unilamellar lipid vesicles in conjunction with small-angle scattering techniques and fluorescence microscopy. We find that lipid phase behaviour remains unperturbed well beyond physiological concentrations of vitamin E (up to a mole fraction of 0.10). Our results are consistent with a proposed line active role of vitamin E at the domain boundary. We discuss the implications of these findings as they pertain to lipid raft modification in native membranes, and propose a new hypothesis for the antioxidant mechanism of vitamin E

Stages of the Demographic Transition from a Child's Perspective: Family Size, Cohort Size, and Children's Resources

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/71989/1/j.1728-4457.2008.00218.x.pd

The Serotonin Transporter Promoter Polymorphism and Childhood Positive and Negative Emotionality

Association studies of the serotonin transporter promoter polymorphism (5-HTTLPR) and negative emotionality (NE) are inconclusive. However, emerging evidence suggests that the association between this polymorphism and NE may be influenced by levels of another temperament trait, positive emotionality (PE). Therefore, this study examined whether the association between the 5-HTTLPR and NE was moderated by PE. A community sample of 413 three-year-old children completed a standardized battery of laboratory tasks designed to tap temperamental emotionality. Children were also genotyped for the 5-HTTLPR. No direct association between 5-HTTLPR genotype and NE was found. However, the interaction of child PE and NE predicted 5-HTTLPR genotype. Furthermore, children with a short allele who were also low in PE had significantly greater NE than children without a short allele or children with high PE. Our findings suggest that the short allele of the 5-HTTLPR is associated with NE only in the context of low PE. Inconsistent links between NE and this gene in previous research may stem from the failure to consider other temperament traits that moderate associations. © 2010 American Psychological Association

Bostonia: The Boston University Alumni Magazine. Volume 9

Founded in 1900, Bostonia magazine is Boston University's main alumni publication, which covers alumni and student life, as well as university activities, events, and programs

Regulation der Genexpression von MYCN in humanen Neuoblastomzellen durch Transkriptionsfaktoren der E2F-Familie

Seit fast 30 Jahren ist bekannt, dass die Amplifikation und Expression des Onkogens MYCN in Neuroblastomen mit einer sehr ungünstigen Prognose für die Patienten einhergeht. Dennoch liegen die Mechanismen der Genregulation von MYCN weiterhin größtenteils im Dunkeln. Die Präsenz potentieller Bindungsstellen für E2F-Proteine im Promotor des MYCN-Gens sowie Zellkulturexperimente lieferten Hinweise auf eine Rolle der Transkriptionsfaktoren der E2F-Familie in der Regulation der N-myc-Expression. Ziel dieser Arbeit war die Beantwortung der Frage, ob E2F-Proteine notwendig sind, um eine primär hohe Expression von N-myc in Neuroblastomzellen mit Amplifikation des Onkogens aufrechtzuerhalten, und ob sie hinreichend sind, um die Transkription von MYCN in Zellen ohne endogene Expression von N-myc einzuleiten. Durch Überexpression des Tumorsuppressorproteins p16, welches zu einer Inaktivierung endogener E2F-Proteine führt, konnte die MYCN-mRNA-Menge in Neuroblastomzellen deutlich gesenkt werden. Vergleichbare Resultate wurden durch Expression von dominant negativem E2F-1 erzielt. Da in einigen Studien gezeigt werden konnte, dass Myc-Proteine ihrerseits E2F-Gene aktivieren können, nehmen wir an, in aggressiven Neuroblastomen könnte eine positive Rückkopplungsschleife zwischen E2F-Transkriptionsfaktoren auf der einen und N-myc auf der anderen Seite existieren, die die gesteigerte Aktivität des MYCN-Onkogens aufrechterhält. Stabil transfizierte E2F-ER-Fusionsproteine waren jedoch nicht in der Lage, das endogene MYCN-Gen in Neuroblastomzellen ohne Expression von N-myc anzuschalten. E2F-Proteine werden folglich für das volle Ausmaß der starken Expression von N-myc in Neuroblastomen benötigt, sind aber nicht ausreichend, um das Onkogen MYCN in Zellen ohne endogenes N-myc zu aktivieren. In der Zukunft könnte durch Verhinderung der Bindung von E2F-Proteinen an den MYCN-Promotor oder durch gentherapeutische Ansätze, die z.B. mittels viraler Infektion den Signalweg zwischen p16 und E2F rekonstruieren, die Expression von N-myc in Neuroblastomen gesenkt werden, so dass die Aggressivität der Tumore reduziert und die individuelle Prognose der Patienten verbessert werden könnte