Panoply of P: An Array of Rhenium-Phosphorus Complexes Generated from a Transition Metal Anion.

We expand upon the synthetic utility of anionic rhenium complex Na[(BDI)ReCp] (1, BDI = N,N'-bis(2,6-diisopropylphenyl)-3,5-dimethyl-ß-diketiminate) to generate several rhenium-phosphorus complexes. Complex 1 reacts in a metathetical manner with chlorophosphines Ph2PCl, MeNHP-Cl, and OHP-Cl to generate XL-type phosphido complexes 2, 3, and 4, respectively (MeNHP-Cl = 2-chloro-1,3-dimethyl-1,3,2-diazaphospholidine; OHP-Cl = 2-chloro-1,3,2-dioxaphospholane). Crystallographic and computational investigations of phosphido triad 2, 3, and 4 reveal that increasing the electronegativity of the phosphorus substituent (C < N < O) results in a shortening and strengthening of the rhenium-phosphorus bond. Complex 1 reacts with iminophosphane Mes*NPCl (Mes* = 2,4,6-tritert-butylphenyl) to generate linear iminophosphanyl complex 5. In the presence of a suitable halide abstraction reagent, 1 reacts with the dichlorophosphine iPr2NPCl2 to afford cationic phosphinidene complex 6+. Complex 6+ may be reduced by one electron to form 6•, a rare example of a stable, paramagnetic phosphinidene complex. Spectroscopic and structural investigations, as well as computational analyses, are employed to elucidate the influence of the phosphorus substituent on the nature of the rhenium-phosphorus bond in 2 through 6. Furthermore, we examine several common analogies employed to understand metal phosphido, phosphinidene, and iminophosphanyl complexes.

Influence of N Terminus Amino Acid on Peptide Cleavage in Solution through Diketopiperazine Formation.

Diketopiperazine (DKP) formation is an important degradation pathway for peptides and proteins. It can occur during synthesis and storage in either solution or the solid state. The kinetics of peptide cleavage through DKP formation have been analyzed for the model peptides Xaa1-Pro2-Gly4-Lys7 [Xaa = Gln, Glu, Lys, Ser, Phe, Trp, Tyr, Cha (ß-cyclohexylalanine), Aib (α-aminoisobutyric acid), Gly, and Val] at multiple elevated temperatures in ethanol with ion mobility spectrometry-mass spectrometry (IMS-MS). When Xaa is an amino acid with a charged or polar side chain, degradation is relatively fast. When Xaa is an amino acid with a nonpolar alkyl side chain, the peptide is relatively stable. For these peptides, a bulky group on the α carbon speeds up dissociation, but the kinetic effects vary in a complicated manner for bulky groups on the ß or γ carbon. Peptides where Xaa has a nonpolar aromatic side chain show moderate dissociation rates. The stability of these peptides is a result of multiple factors. The reaction rate is enhanced by (1) the stabilization of the late transition state through the interaction of an aromatic ring with the nascent DKP ring or lowering the activation energy of nucleophilic attack intermediate state through polar or charged residues and (2) the preference of the cis proline bond favored by the aromatic N-terminus. The number of unseen intermediates and transition state thermodynamic values are derived for each peptide by modeling the kinetics data. Most of the transition states are entropically favored (ΔS⧧ ∼ -5 to +31 J·mol-1·K-1), and all are enthalpically disfavored (ΔH⧧ ∼ 93 to 109 kJ·mol-1). The Gibbs free energy of activation is similar for all of the peptides studied here (ΔG⧧ ∼ 90-99 kJ·mol-1).

Diketopiperazine Formation from FPGnK (n = 1-9) Peptides: Rates of Structural Rearrangements and Mechanisms.

Peptides with penultimate proline residues undergo trans → cis isomerization of the Phe1-Pro2 peptide bond followed by spontaneous bond cleavage at the Pro2-Xxx3 bond (where Xxx is another amino acid residue), leading to cleavage of the Pro2-Xxx3 bond and formation of a diketopiperazine (DKP). In this paper, ion mobility spectrometry and mass spectrometry techniques were used to study the dissociation kinetics of nine peptides [Phe1-Pro2-Glyn-Lysn+3 (n = 1-9)] in ethanol. Shorter (n = 1-3) peptides are found to be more stable than longer (n = 4-9) peptides. Alanine substitution studies indicate that, when experiments are initiated, the Phe1-Pro2 bond of the n = 9 peptide exists exclusively in the cis configuration, while the n = 1-8 peptides appear to exist initially with both cis- and trans-Phe1-Pro2 configured bonds. Molecular dynamics simulations indicate that intramolecular hydrogen bonding interactions stabilize conformations of shorter peptides, thus inhibiting DKP formation. Similar stabilizing interactions appear less frequently in longer peptides. In addition, in smaller peptides, the N-terminal amino group is more likely to be charged compared to the same group in longer peptides, which would inhibit the dissociation through the DKP formation mechanism. Analysis of temperature-dependent kinetics measurements provides insight about the mechanism of bond cleavage. The analysis gives the following transition state thermochemistry: ΔG⧧ values range from 94.6 ± 0.9 to 101.5 ± 1.9 kJ·mol-1, values of ΔH⧧ range from 89.1 ± 0.9 to 116.7 ± 1.5 kJ·mol-1, and ΔS⧧ values range from -25.4 ± 2.6 to 50.8 ± 4.2 J·mol-1·K-1. Proposed mechanisms and thermochemistry are discussed.

Solid-state packing dictates the unexpected solubility of aromatic peptides.

The understanding and prediction of the solubility of biomolecules, even of the simplest ones, reflect an open question and unmet need. Short aromatic tripeptides are among the most highly aggregative biomolecules. However, in marked contrast, Ala-Phe-Ala (AFA) was surprisingly found to be non-aggregative and could be solubilized at millimolar concentrations. Here, aiming to uncover the underlying molecular basis of its high solubility, we explore in detail the solubility, aggregation propensity, and atomic-level structure of the tripeptide. We demonstrate an unexpectedly high water solubility of AFA reaching 672 mM, two orders of magnitude higher than reported previously. The single crystal structure reveals an anti-parallel ß sheet conformation devoid of any aromatic interactions. This study provides clear mechanistic insight into the structural basis of solubility and suggests a simple and feasible tool for its estimation, bearing implications for design of peptide drugs, peptides materials, and advancement of peptide nanotechnology.

Influence of Solvents upon Diketopiperazine Formation of FPG8K.

Ion mobility spectrometry (IMS) and mass spectrometry (MS) techniques were used to monitor diketopiperazine (DKP) formation from the peptide FPG8K at multiple defined temperatures in methanol, ethanol, propanol, and water, with the motivation to study the effect of solvent polarity on spontaneous solution dissociation. The reaction rate increases with decreasing solvent polarity. The observed rates of trans → cis isomerization of Phe1-Pro2 and the cis-Pro2 isomer dissociation result in the cis isomer growing in abundance relative to the trans isomer throughout the reaction in all solvents. Analysis of rate constants derived from the data using a sequential unimolecular kinetics model that includes hidden intermediate states yields transition state thermodynamic values for both trans → cis isomerization of Phe1-Pro2 and dissociation. The measured thermochemistry appears to be closely correlated with these solvents' dielectric constants: a lower solvent dielectric constant accelerates the reaction by reducing the enthalpic barrier, albeit with slight entropic restriction.

A multicountry European study on Succeed: a general quality improvement tool in HIV prevention.

The context-sensitive nature of the European HIV epidemic (e.g. differences in key populations, prevention settings, resource commitments) makes it challenging to monitor and evaluate HIV prevention and sexual health promotion programs. Systematic quality improvement (QI) tools and quality indicators adapted to HIV prevention are not widely known or implemented. The European 'Quality Action' introduced five such tools in 26 countries involving 45 nongovernmental and governmental organizations during 2014-2016. Approximately, half of the organizations used the tool 'Succeed'. This study aims to describe challenges and supportive factors in its use, and answers the research question if Succeed can be applied as a general QI tool despite the high contextuality of the European HIV prevention field. Mixed method research was conducted: data from quantitative pre- and post-training and implementation surveys were triangulated with qualitative data from multiple data sources analysed inductively. In a second analytical step, Chaudoir's evidence-based framework on implementation of innovations (2013) guided the data analysis on five levels: policy, organization, functionality of the tool, results and perceived innovations. Succeed contributed to goal and result orientation, facilitated stakeholders' participation and contributed to organizational development. Succeed was used in similar ways and with similar results across different policy contexts, types of organizations, target groups and interventions. Contributing factors for sustainable implementation were supporting environments, sufficient resources and a strategy for training tool users. Findings strongly support the use of Succeed as a general QI tool to monitor, document, adapt and improve HIV prevention and sexual health promotion work across Europe.

Evidence for Many Unique Solution Structures for Chymotrypsin Inhibitor 2: A Thermodynamic Perspective Derived from vT-ESI-IMS-MS Measurements.

Chymotrypsin inhibitor 2 (CI-2) is a classic model for two-state cooperative protein folding and is one of the most extensively studied systems. Alan Fersht, a pioneer in the field of structural biology, has studied the wild-type (wt) and over 100 mutant forms of CI-2 with traditional analytical and biochemical techniques. Here, we examine wt CI-2 and three mutant forms (A16G, K11A, L32A) to demonstrate the utility of variable-temperature (vT) electrospray ionization (ESI) paired with ion mobility spectrometry (IMS) and mass spectrometry (MS) to map the free energy folding landscape. As the solution temperature is increased, the abundance of each of the six ESI charge states for wt CI-2 and each mutant is found to vary independently. These results require that at least six unique types of CI-2 solution conformers are present. Ion mobility analysis reveals that within each charge state there are additional conformers having distinct solution temperature profiles. A model of the data at ∼30 different temperatures for all four systems suggests the presence of 41 unique CI-2 solution conformations. A thermodynamic analysis of this system yields values of ΔCp as well as ΔG, ΔH, and ΔS for each state at every temperature studied. Detailed energy landscapes derived from these data provide a rare glimpse into Anfinsen's thermodynamic hypothesis and the process of thermal denaturation, normally thought of as a cooperative two-state transition involving the native state and unstructured denatured species. Specifically, as the temperature is varied, the entropies and enthalpies of different conformers undergo dramatic changes in magnitude and relative order to maintain the delicate balance associated with equilibrium.

Monitoring the stabilities of a mixture of peptides by mass-spectrometry-based techniques.

Biomolecular degradation plays a key role in proteostasis. Typically, proteolytic enzymes degrade proteins into smaller peptides by breaking amino acid bonds between specific residues. Cleavage around proline residues is often missed and requires highly specific enzymes for peptide processing due to the cyclic proline side-chain. However, degradation can occur spontaneously (i.e. in the absence of enzymes). In this study, the influence of the first residue on the stability of a series of penultimate proline containing peptides, with the sequence Xaa-Pro-Gly-Gly (where Xaa is any amino acid), is investigated with mass spectrometry techniques. Peptides were incubated as mixtures at various solution temperatures (70℃ to 90℃) and were periodically sampled over the duration of the experiment. At elevated temperatures, we observe dissociation after the Xaa-Pro motif for all sequences, but at different rates. Transition state thermochemistry was obtained by studying the temperature-dependent kinetics and although all peptides show relatively small differences in the transition state free energies (∼95 kJ/mol), there is significant variability in the transition state entropy and enthalpy. This demonstrates that the side-chain of the first amino acid has a significant influence on the stability of the Xaa-Pro sequence. From these data, we demonstrate the ability to simultaneously measure the dissociation kinetics and relative transition state thermochemistries for a mixture of peptides, which vary only in the identity of the N-terminal amino acid.

Solvent Mediation of Peptide Conformations: Polyproline Structures in Water, Methanol, Ethanol, and 1-Propanol as Determined by Ion Mobility Spectrometry-Mass Spectrometry.

Ion mobility spectrometry and circular dichroism spectroscopy are used to examine the populations of the small model peptide, polyproline-13 in water, methanol, ethanol, and 1-propanol over a range of solution temperatures (from 288 to 318 K). At low temperatures, the less-polar solvents (1-propanol and ethanol) favor the all-cis polyproline I helix (PPI); as the temperature is increased, the trans-configured polyproline II helix (PPII) is formed. In polar solvents (methanol and water), PPII is favored at all temperatures. From the experimental data, we determine the relative stabilities of the eight structures in methanol, ethanol, and 1-propanol, as well as four in water, all with respect to PPII. Although these conformers show relatively small differences in free energies, substantial variability is observed in the enthalpies and entropies across the structures and solvents. This requires that enthalpies and entropies be highly correlated: in 1-propanol, cis-configured PPI conformations are energetically favorable but entropically disfavored. In more polar solvents, PPI is enthalpically less favorable and entropy favors trans-configured forms. While either ΔH0 or ΔS0 can favor different structures, no conformation in any solvent is simultaneously energetically and entropically stabilized. These data present a rare opportunity to examine the origin of conformational stability. Graphical Abstract ᅟ.

Substantial Heterogeneity in Progress Toward Reaching the 90-90-90 HIV Target in the WHO European Region.

BACKGROUND: Achieving the UNAIDS 90-90-90 target by 2020 is expected to end the HIV epidemic by 2030. We report on progress in the WHO European Region in meeting this target. METHODS: The European Centre for Disease Prevention and Control (ECDC) sent questionnaires to 55 countries in 2016. We report estimates for 4 stages of the continuum of HIV care (living with HIV, diagnosed, treated, and virally suppressed), corresponding to the Joint United Nations Programme on HIV and AIDS (UNAIDS) target and explore differences by subregion and challenges with reporting data. FINDINGS: Forty-four countries provided data for ≥1 stage, and 29 for all 4 stages. Estimated HIV prevalence was 0.19% (range 0.02%-0.84%, n = 37 countries providing stage 1 data). The proportion diagnosed of people living with HIV ranged from 38% to 98% (n = 37 reporting number of people living with HIV and diagnosed). The proportion on ART of those diagnosed ranged from 27% to 96% (n = 40 reporting numbers diagnosed and treated), and viral suppression rates ranged from 32% to 97% (n = 31 providing numbers treated and virally suppressed). The overall continuum of care estimate for 29 countries with complete data was 81-84-88, which differed by subregion: 84-88-90, 84-69-62, and 57-45-57 for the western, central, and eastern subregions, respectively. Challenges in reporting data included absence of a single data source for all stages, shortage of expertise, and lack of financial and human resources. CONCLUSIONS: There is an urgent need to strengthen HIV testing programs throughout Europe, particularly in the eastern subregion, and to remove constraints hampering access to testing and care. Recent changes to treatment guidelines should help reduce the numbers diagnosed not treated.

Predicting Cancer-Prevention Behavior: Disentangling the Effects of Risk Aversion and Risk Perceptions.

Experimental and survey research spanning the last two decades concludes that people who are more risk tolerant are more likely to engage in risky health activities such as smoking and heavy alcohol consumption, and are more likely to be obese. Subjective perceptions of the risk associated with different activities have also been found to be associated with health behaviors. While there are numerous studies that link risk perceptions with risky behavior, it is notable that none of these controls for risk aversion. Similarly, studies that control for risk aversion fail to control for risk misperceptions. We use a survey of 474 men and women to investigate the influence of risk aversion, risk misperceptions, and cognitive ability on the choice to engage in behaviors that either increase or mitigate cancer risk. We measure optimism in two dimensions: baseline optimists are those who inaccurately believe their cancer risk to be below its expert-assessed level, while control optimists are those who believe they can reduce their risk of cancer (by changing their lifestyle choices) to a greater extent than is actually the case. Our results indicate that baseline optimism is significantly and negatively correlated with subjects' tendencies to engage in cancer-risk-reducing behaviors, and positively correlated with risky behaviors. Subjects' control misperceptions also appear to play a role in their tendency to engage in risky and prevention behaviors. When controlling for both of these types of risk misperception, risk aversion plays a much smaller role in determining health behaviors than found in past studies.

Long-Lived Intermediates in a Cooperative Two-State Folding Transition.

Biomolecular folding often occurs through a cooperative two-state reactant ↔ product transition; the term cooperative does not convey that intermediate structures are nonexistent but rather that these states are not observable by existing experimental techniques. Because of this, few intermediates have been studied and characterized. Recently, ion mobility spectrometry (IMS) measurements revealed that the oligomer polyproline-13 (Pro13, which in propanol (PrOH) favors the right-handed helical PPI structure having adjacent pyrrolidine rings in a cis configuration) folds through six sequential long-lived intermediates as it converts to the all-trans-configured PPII structure that is favored in aqueous solutions. Here, we examine the PPIPrOH → PPIIaq folding transition for a HisPro13 sequence, i.e., Pro13 having a single histidine residue added to the N-terminus. Remarkably, the IMS measurements show that, upon addition of histidine, all of the IMS peaks associated with intermediate structures disappear. Instead, HisPro13 folds via a cooperative two-state transition, delayed by a significant induction period. The induction period is temperature dependent-shifting the transition to longer times at lower temperatures. Equilibrium studies show that the HisPro13 PPIPrOH → PPIIaq transition is endothermic but favored entropically. From these clues, we propose a sequential folding mechanism and develop a model that suggests that ∼13-17 long-lived intermediates are likely responsible for the induction period. In this model, intermediates are separated by average individual activation barriers of ∼90 kJ·mol-1, and are entropically favorable.

Ion-molecule chemistry within boron tribromide clusters: experiment and theory.

Molecular clusters of BBr3 were subjected to electron ionization and mass analysis in a reflectron time-of-flight mass spectrometer. Five series of cluster ions were observed, with formulas corresponding to each of the possible fragment ions of BBr3 being solvated by neutral BBr3 molecules. Geometry optimizations on the observed cluster ions using density functional theory (B3LYP/6-31G*) predict that fragment ions smaller than BBr3+ undergo reactions with neutral BBr3 molecules to form covalently bound adduct species that function as core ions within the clusters. Once all boron atoms are saturated, the reactions cease, and larger cluster ions consist of BBr3 molecules loosely bound to the core ions. Divalent bromine atoms are present in at least three of the cluster ions, and most of the intermolecular contact within the clusters is between Br atoms. Enthalpies of formation, addition reactions, and BBr3 elimination from the cluster ions were derived from B3LYP and MP2 calculations at the B3LYP/6-31G* geometries using both the 6-31G* and the 6-311++G(2df,2p) basis sets. The results are compared to limiting expectations based on known bulk thermochemistry.

Optical spectroscopy of RuC: 18,000-24,000 cm(-1).

The optical spectrum of diatomic RuC has been recorded from 17 800 to 24 200 cm(-1). Three previously unidentified excited electronic states were analyzed and identified as having Omega' = 0, Omega' = 2, and Omega' = 3. The Omega' = 3 state was determined to be a 3Delta3 state that is suggested to arise from a mixture of the 10sigma(2)11sigma(2)5pi(3)2delta(3)12sigma(1)6pi(1) and 10sigma(2)11sigma(1)5pi(3)2delta(3)12sigma(2)6pi(1) electronic configurations. Three additional bands belonging to the previously observed [18.1] (1)Pi<--X (1)Sigma(+) system were analyzed to obtain B(e) (')=0.558 244(48) cm(-1), alpha(e) (')=0.004 655(27) cm(-1), omegae' = 887.201(37) cm(-1), and omega(e) 'xe' = 5.589(7) cm(-1) for the 102Ru 12C isotopomer (1sigma error limits). A Rydberg-Klein-Rees analysis was then performed using the determined spectroscopic constants of the [18.1] 1Pi state, and similar analyses were performed for the previously observed states. The resulting potential energy curves are provided for the 100Ru 12C, 101Ru 12C, 102Ru 12C, and 104Ru 12C isotopic species.