Lapatinib-binding protein kinases in the African trypanosome: identification of cellular targets for kinase-directed chemical scaffolds.

Human African trypanosomiasis is caused by the eukaryotic microbe Trypanosoma brucei. To discover new drugs against the disease, one may use drugs in the clinic for other indications whose chemical scaffolds can be optimized via a medicinal chemistry campaign to achieve greater potency against the trypanosome. Towards this goal, we tested inhibitors of human EGFR and/or VEGFR as possible anti-trypanosome compounds. The 4-anilinoquinazolines canertinib and lapatinib, and the pyrrolopyrimidine AEE788 killed bloodstream T. brucei in vitro with GI(50) in the low micromolar range. Curiously, the genome of T. brucei does not encode EGFR or VEGFR, indicating that the drugs recognize alternate proteins. To discover these novel targets, a trypanosome lysate was adsorbed to an ATP-sepharose matrix and washed with a high salt solution followed by nicotinamide adenine dinucleotide (NAD(+)). Proteins that remained bound to the column were eluted with drugs, and identified by mass spectrometry/bioinformatics. Lapatinib bound to Tb927.4.5180 (termed T. brucei lapatinib-binding protein kinase-1 (TbLBPK1)) while AEE788 bound Tb927.5.800 (TbLBPK2). When the NAD(+) wash was omitted from the protocol, AEE788, canertinib and lapatinib eluted TbLBPK1, TbLBPK2, and Tb927.3.1570 (TbLBPK3). In addition, both canertinib and lapatinib eluted Tb10.60.3140 (TbLBPK4), whereas only canertinib desorbed Tb10.61.1880 (TbCBPK1). Lapatinib binds to a unique conformation of protein kinases. To gain insight into the structural basis for lapatinib interaction with TbLBPKs, we constructed three-dimensional models of lapatinib•TbLBPK complexes, which confirmed that TbLBPKs can adopt lapatinib-compatible conformations. Further, lapatinib, AEE788, and canertinib were docked to TbLBPKs with favorable scores. Our studies (a) present novel targets of kinase-directed drugs in the trypanosome, and (b) offer the 4-anilinoquinazoline and pyrrolopyrimidines as scaffolds worthy of medicinal chemistry and structural biology campaigns to develop them into anti-trypanosome drugs

Photoexcitation of iodide ion-pyrimidine clusters above the electron detachment threshold : Intracluster electron transfer versus nucleobase-centred excitations

Laser photodissociation spectroscopy of the I-·thymine (I-·T) and I-·cytosine (I-·C) nucleobase clusters has been conducted for the first time across the regions above the electron detachment thresholds to explore the excited states and photodissociation channels. Although photodepletion is strong, only weak ionic photofragment signals are observed, indicating that the clusters decay predominantly by electron detachment. The photodepletion spectra of the I-·T and I-·C clusters display a prominent dipole-bound excited state (I) in the vicinity of the vertical detachment energy (∼4.0 eV). Like the previously studied I-·uracil (I-·U) cluster [W. L. Li et al., J. Chem. Phys. 145, 044319 (2016)], the I-·T cluster also displays a second excited state (II) centred at 4.8 eV, which we similarly assign to a π-π∗ nucleobase-localized transition. However, no distinct higher-energy absorption bands are evident in the spectra of the I-·C. Time-dependent density functional theory (TDDFT) calculations are presented, showing that while each of the I-·T and I-·U clusters displays a single dominant π-π∗ nucleobase-localized transition, the corresponding π-π∗ nucleobase transitions for I-·C are split across three separate weaker electronic excitations. I- and deprotonated nucleobase anion photofragments are observed upon photoexcitation of both I-·U and I-·T, with the action spectra showing bands (at 4.0 and 4.8 eV) for both the I- and deprotonated nucleobase anion production. The photofragmentation behaviour of the I-·C cluster is distinctive as its I- photofragment displays a relatively flat profile above the expected vertical detachment energy. We discuss the observed photofragmentation profiles of the I-·pyrimidine clusters, in the context of the previous time-resolved measurements, and conclude that the observed photoexcitations are primarily consistent with intracluster electron transfer dominating in the near-threshold region, while nucleobase-centred excitations dominate close to 4.8 eV. TDDFT calculations suggest that charge-transfer transitions [Iodide n (5p6) → Uracil σ∗] may contribute to the cluster absorption profile across the scanned spectral region, and the possible role of these states is also discussed

Creative women in Peru: outliers in a machismo world

Gender segregation begins early and is reinforced within the workplace. Advertising creative departments appear to have extreme gender segregation with women representing just 20% of all those working within creative departments worldwide. Yet, creativity does not depend on gender. Thus, the underrepresentation of women is particularly troubling. In Peru women comprise 3% to 10.4% of all people working in advertising creative, which suggests the situation for creative women in Peru is dire. In order to understand this phenomenon, and with the hope of finding solutions, this study uses indepth interviews to explore the experiences of Peruvian women working in advertising creative departments. The study investigates three primary aspects of Peruvian creative women’s experiences. First, it looks at relationships with colleagues and clients. Second, work/life balance is explored. Third, the study examines how the environment within creative departments constrains creative women’s employment and advancement opportunities. Findings suggest that Peruvian creative departments are strongly machismo environments where discrimination and gender segregation are staunchly entrenched. This machismo environment creates challenging relationships between creative women and their colleagues and clients, it negatively impacts creative women’s work/life balance and it leads to severely constrained hiring, promotion and retention of creative women in Peruvian advertising agencies. The discussion closes with suggestions to help creative women succeed in Peruvian creative departments

A transverse current rectification in graphene superlattice

A model for energy spectrum of superlattice on the base of graphene placed on the striped dielectric substrate is proposed. A direct current component which appears in that structure perpendicularly to pulling electric field under the influence of elliptically polarized electromagnetic wave was derived. A transverse current density dependence on pulling field magnitude and on magnitude of component of elliptically polarized wave directed along the axis of a superlattice is analyzed.Comment: 12 pages, 6 figure

Evaluation of the MagicplexTM sepsis real-time test for the rapid diagnosis of bloodstream infections in adults

Sepsis is a serious health condition worldwide, affecting more than 30 million people globally each year. Blood culture (BC) is generally used to diagnose sepsis because of the low quantity of microbes occurring in the blood during such infections. However, ~50% of bloodstream infections (BSI) give negative BC, this figure being higher for sepsis, which delays the start of appropriate antimicrobial therapy. This prospective study evaluated a multiplex real-time polymerase chain reaction, the MagicplexTM Sepsis test (MP), for the detection of pathogens from whole blood, comparing it to routine BC. We analyzed 809 blood samples from 636 adult patients, with 132/809 (16.3%) of the samples positive for one or more relevant microorganism according to BC and/or MP. The sensitivity and specificity of MP were 29 and 95%, respectively, while the level of agreement between BC and MP was 87%. The rate of contaminated samples was higher for BC (10%) than MP (4.8%) (P < 0.001). Patients with only MP-positive samples were more likely to be on antimicrobial treatment (47%) than those with only BC-positive samples (18%) (P = 0.002). In summary, the MP test could be useful in some clinical setting, such as among patients on antibiotic therapy. Nevertheless, a low sensitivity demonstrated impairs its use as a part of a routine diagnostic algorithm