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Glucagon-like peptide-1 (GLP-1) lowers blood glucose by inducing insulin but also has other poorly understood properties. Here, we show that hydroxy amino acids (Thr11, Ser14, Ser17, Ser18) in GLP-1(7-36) act in concert to direct cell signaling. Mutating any single residue to alanine removes one hydroxyl group, thereby reducing receptor affinity and cAMP 10-fold, with Ala11 or Ala14 also reducing β-arrestin-2 10-fold, while Ala17 or Ala18 also increases ERK1/2 phosphorylation 5-fold. Multiple alanine mutations more profoundly bias signaling, differentially silencing or restoring one or more signaling properties. Mutating three serines silences only ERK1/2, the first example of such bias. Mutating all four residues silences β-arrestin-2, ERK1/2, and Ca

The actin cytoskeleton is a potentially vulnerable property of cancer cells, yet chemotherapeutic targeting attempts have been hampered by unacceptable toxicity. In this study, we have shown that it is possible to disrupt specific actin filament populations by targeting isoforms of tropomyosin, a core component of actin filaments, that are selectively upregulated in cancers. A novel class of anti-tropomyosin compounds has been developed that preferentially disrupts the actin cytoskeleton of tumor cells, impairing both tumor cell motility and viability. Our lead compound, TR100, is effective in vitro and in vivo in reducing tumor cell growth in neuroblastoma and melanoma models. Importantly, TR100 shows no adverse impact on cardiac structure and function, which is the major side effect of current anti-actin drugs. This proof-of-principle study shows that it is possible to target specific actin filament populations fundamental to tumor cell viability based on their tropomyosin isoform composition. This improvement in specificity provides a pathway to the development of a novel class of anti-actin compounds for the potential treatment of a wide variety of cancers. Cancer Res; 73(16); 5169–82. ©2013 AACR.

PDF file - 1137K, Supplementary Figure 1: Tropomyosin siRNA knockdown is isoform specific and does not result in any compensation. Supplementary Figure 2: TR100 nullifies the impact of Tm5NM1 on actin filament depolymerization kinetics. Supplementary Figure 3: Anti-Tropomyosin compound impact on the actin cytoskeleton assessed by the microfilament disruption assay. Supplementary Figure 4: TR100 targets the actin cytoskeleton and preferentially disrupts LMW-cytoskeletal tropomyosin containing filaments. Supplementary Figure 5: TR100 shows no overt impact on synaptic integrity of hippocampal neurons. Supplementary Figure 6: TR100 induces apoptosis in tumor cells via a mitochondrial pathway. Supplementary Figure 7: TR100 does not impact liver function in vivo.Supplementary Table 1: Summary of impact of TR100 on tumor cell viability.

Rationale Stress exposure during adolescence contributes to developing a methamphetamine (METH) use disorder. However, most of the studies investigating addiction-related behaviours include only male rodents, despite METH addiction rates being higher in females. Furthermore, animal studies investigating the effects of stress on methamphetamine addiction have used only basic self-administration models which may not be sensitive to the effects of stress. Objectives This project explored whether adolescent isolation stress exposure increases the incidence of four key addiction-related behaviours in female rats. Methods Thirty-two female rat pups were caged in groups of four or individually during adolescence from postnatal (PND) day 22, with the latter being re-socialised in groups of four on PND 43. In adulthood, rats were tested for addiction-like behaviours in a METH self-administration paradigm modelling motivation to take METH, persistence in drug-seeking behaviour when METH was not available, resistance to extinction, and propensity to reinstate after a period of withdrawal. Results Adolescent social isolation resulted in lower METH intake during acquisition; however, the paradigm modelling drug-seeking when the drug was unavailable engendered intermittent METH bingeing in all rats, abolishing the group differences in intake during this phase. Adolescent social isolation also accelerated extinction of non-reinforced lever pressing, and increased stress-primed reinstatement, compared to the group-housed rats. Conclusions Adolescent social isolation stress alters various methamphetamine addiction-like behaviours in female rats.

Cyclic peptides with esters but not N-methyl amides are the smallest known alpha helices in water and can enter cells.

There is high potential for ecosystem restoration across tropical savannah-dominated regions, but the benefits that could be gained from this restoration are rarely assessed. This study focuses on the Brazilian Cerrado, a highly species-rich savannah-dominated region, as an exemplar to review potential restoration benefits using three metrics: net biomass gains, plant species richness and ability to connect restored and native vegetation. Localized estimates of the most appropriate restoration vegetation type (grassland, savannah, woodland/forest) for pasturelands are produced. Carbon sequestration potential is significant for savannah and woodland/forest restoration in the seasonally dry tropics (net biomass gains of 58.2 ± 37.7 and 130.0 ± 69.4 Mg ha −1 ). Modelled restoration species richness gains were highest in the central and south-east of the Cerrado for savannahs and grasslands, and in the west and north-west for woodlands/forests. The potential to initiate restoration projects across the whole of the Cerrado is high and four hotspot areas are identified. We demonstrate that landscape restoration across all vegetation types within heterogeneous tropical savannah-dominated regions can maximize biodiversity and carbon gains. However, conservation of existing vegetation is essential to minimizing the cost and improving the chances of restoration success. This article is part of the theme issue ‘Understanding forest landscape restoration: reinforcing scientific foundations for the UN Decade on Ecosystem Restoration’.

An alpha helical turn can be reproduced in a cyclic pentapeptide if the first and fifth amino acid sidechains are correctly joined. Here structural studies (CD, NMR

N- or C-methylation in natural and synthetic cyclic peptides can increase membrane permeability, but it remains unclear why this happens in some cases but not others. Here we compare three-dimensional structures for cyclic peptides from six families, including isomers differing only in the location of an N- or Cα-methyl substituent. We show that a single methyl group only increases membrane permeability when it connects or expands hydrophobic surface patches. Positional isomers, with the same molecular weight, hydrogen bond donors/acceptors, rotatable bonds, calculated LogP, topological polar surface area, and total hydrophobic surface area, can have different membrane permeabilities that correlate with the size of the largest continuous hydrophobic surface patch. These results illuminate a key local molecular determinant of membrane permeability.

Millimeter-wave lens antennas will be essential for future wireless access. Conventionally, they increase the gain in the boresight direction only. In this article, cascaded Fresnel zone plate lenses are combined with a phased array to increase the gain at wide steering angles of ±52°. The side lenses are tilted to align with the maximum steering angle and cascaded to increase the focusing gain. The inner lenses increase the gain by 2.45 dB at boresight, and by 3.19 dB at the maximum steering angle. When the side lenses are repositioned, the simulated focusing gain increases to 4.69 dB. Asymmetric amplitude distributions are proposed to prevent the main lobe from splitting. An eight-element seven-lens prototype operating at 28 GHz achieved a gain from 12.96 to 15.35 dBi with a bandwidth of at least 1.3 GHz for all measured beam directions. The maximum measured azimuthal beamwidth was 27°. A design procedure and a theoretical analysis of diffraction through the lenses are provided. By increasing the SNR, this beamforming antenna could improve the coverage of three-sector 5G microcell base stations, and support gigabit wireless links for vehicular, rail, and satellite communications.

In its 33 years, ADDR has published regularly on the po5tential of oral delivery of biologics especially peptides and proteins. In the intervening period, analysis of the preclinical and clinical trial failures of many purported platform technologies has led to reflection on the true status of the field and reigning in of expectations. Oral formulations of semaglutide, octreotide, and salmon calcitonin have completed Phase III trials, with oral semaglutide being approved by the FDA in 2019. The progress made with oral peptide formulations based on traditional permeation enhancers is against a background of low and variable oral bioavailability values of ~1%, leading to a current perception that only potent peptides with a viable cost of synthesis can be realistically considered. Desirable features of candidates should include a large therapeutic index, some stability in the GI tract, a long elimination half-life, and a relatively low clearance rate. Administration in nanoparticle formats have largely disappointed, with few prototypes reaching clinical trials: insufficient particle loading, lack of controlled release, low epithelial particle uptake, and lack of scalable synthesis being the main reasons for discontinuation. Disruptive technologies based on engineered devices promise improvements, but scale-up and toxicology aspects are issues to address. In parallel, medicinal chemists are synthesizing stable hydrophobic macrocyclic candidate peptides of lower molecular weight and with potential for greater oral bioavailability than linear peptides, but perhaps without the same requirement for elaborate drug delivery systems. In summary, while there have been advances in understanding the limitations of peptides for oral delivery, low membrane permeability, metabolism, and high clearance rates continue to hamper progress.

The introduction of an amide bond linking side chains of the first and fifth amino acids forms a cyclic pentapeptide that optimally stabilizes the smallest known α-helix in water. The origin of the stabilization is unclear. The observed dependence of α-helicity on the solvent and cyclization linker led us to discover a novel long-range n to π* interaction between a main-chain amide oxygen and a uniquely positioned carbonyl group in the linker of cyclic pentapeptides. CD and NMR spectra, NMR and X-ray structures, modelling, and MD simulations reveal that this first example of a synthetically incorporated long-range n to π* CO⋅⋅⋅Cγ =Ο interaction uniquely enforces an almost perfect and remarkably stable peptide α-helix in water but not in DMSO. This unusual interaction with a covalent amide bond outside the helical backbone suggests new approaches to synthetically stabilize peptide structures in water.

Description, Novel macrocyclic peptides are found that bind and modulate the function of the retromer membrane trafficking complex., The retromer complex (Vps35-Vps26-Vps29) is essential for endosomal membrane trafficking and signaling. Mutation of the retromer subunit Vps35 causes late-onset Parkinson’s disease, while viral and bacterial pathogens can hijack the complex during cellular infection. To modulate and probe its function, we have created a novel series of macrocyclic peptides that bind retromer with high affinity and specificity. Crystal structures show that most of the cyclic peptides bind to Vps29 via a Pro-Leu–containing sequence, structurally mimicking known interactors such as TBC1D5 and blocking their interaction with retromer in vitro and in cells. By contrast, macrocyclic peptide RT-L4 binds retromer at the Vps35-Vps26 interface and is a more effective molecular chaperone than reported small molecules, suggesting a new therapeutic avenue for targeting retromer. Last, tagged peptides can be used to probe the cellular localization of retromer and its functional interactions in cells, providing novel tools for studying retromer function.

Protein–protein interactions involve hotspots as small as 4 sequential amino acids. Corresponding tetrapeptides have no structure in water. Here we report linking side chains of amino acids X and Z to form 24 cyclic tetrapeptides, cyclo-[XAAZ]-NH2, and stabilise 14–18 membered rings that mimic different kinds of non-regular secondary structures found in protein hotspots. 2D NMR spectra allowed determination of 3D structures for 14 cyclic tetrapeptides in water. Five formed two (i, i + 3) hydrogen bonds and a beta/gamma (6, 7) or beta (9, 19, 20) turn; eight formed one (i, i + 4) hydrogen bond and twisted into a non-helical (13, 18, 21, 22, 24) or helical (5, 17, 23) alpha turn; one was less structured (15). A beta or gamma turn was favoured for Z = Dab, Orn or Glu due to a χ1 gauche (+) rotamer, while an alpha turn was favoured for Z = Dap (but not X = Dap) due to a gauche (−) rotamer. Surprisingly, an unstructured peptide ARLARLARL could be twisted into a helix when either a helical or non-helical alpha turn (5, 13, 17, 18, 21–24) with Z = Dap was attached to the N-terminus. These structural models provide insights into stability for different turns and twists corresponding to non-regular folds in protein hotspots.

Agroforestry is an integration of trees in cropland or grassland and is discussed, within Germany and the EU, as a potential “Green Solution” for agriculture. Agroforestry alters the microclimate, productivity, biodiversity, and nutrient and water usage – as compared to standard agricultural practise. A potentially key benefit is the higher carbon sequestration of agroforestry, relative to monoculture systems, which could provide an interesting option for mitigating climate change, while still providing valuable arable land. Net ecosystem exchange studies of CO2 (NEE) of agroforestry systems are rare, in comparison to the extensive studies of NEE of agricultural systems (croplands and grasslands). Therefore, the current study, as part of the SIGNAL (sustainable intensification of agriculture through agroforestry) project, investigates the NEE of agroforestry compared to that of monoculture agriculture.At five locations across Germany, paired flux measurements above agroforestry and monoculture agronomy are performed using innovative low-cost CO2 eddy covariance sensors (slow response Vaisala GMP343 IRGA, with custom made housing). During the summer of 2020 simultaneous measurements of the low-cost setup and a LI-COR 7200 are performed, above grassland at 3.5 m and adjacent agroforestry grassland at 10 m measurements height.The low-cost eddy covariance system is able to capture the turbulent (diurnal) CO2 flux dynamics and the response to management activities. After spectral corrections and applying quality control, the low-cost system at the agroforestry site (slope = 0.92, R2 = 0.88) performs better than the low-cost system at the grassland site (slope = 0.67, R2 = 0.80), when compared to the LI-COR measurements. This is probably due to the difference in turbulence caused by different surface roughness and measurement height. The preliminary cumulative carbon flux during the four-month measurement campaign shows a significant difference between the grassland (source of (+) 16-38 gC/m2) and agroforestry grassland (sink of (-) 148-164 gC/m2), in favour of agroforestry. By applying post processing software, we aim to further optimize the frequency corrections for the low-cost system. In the future the obtained post processing scheme will be applied to the other low-cost eddy covariance systems within the project.

The Retromer complex (Vps35-Vps26-Vps29) is essential for endosomal membrane trafficking and signalling. Mutations in Retromer cause late-onset Parkinson’s disease, while viral and bacterial pathogens can hijack the complex during cellular infection. To modulate and probe its function we have created a novel series of macrocyclic peptides that bind Retromer with high affinity and specificity. Crystal structures show the majority of cyclic peptides bind to Vps29 via a Pro-Leu-containing sequence, structurally mimicking known interactors such as TBC1D5, and blocking their interaction with Retromer in vitro and in cells. By contrast, macrocyclic peptide RT-L4 binds Retromer at the Vps35-Vps26 interface and is a more effective molecular chaperone than reported small molecules, suggesting a new therapeutic avenue for targeting Retromer. Finally, tagged peptides can be used to probe the cellular localisation of Retromer and its functional interactions in cells, providing novel tools for studying Retromer function.

Funding: The research was carried out as part of a project funded by the Malaysian Palm Oil Board (MPOB). L. K. K and E. R. are both employees of MPOB. The research was carried out with the support of Sarawak Oil Palm Berhard (SOPB) on whose land the research project was based. Need for regional economic development and global demand for agro‐industrial commodities has resulted in large‐scale conversion of forested landscapes to industrial agriculture across South East Asia. However, net emissions of CO2 from tropical peatland conversions may be significant and remain poorly quantified, resulting in controversy around the magnitude of carbon release following conversion. Here we present long term, whole ecosystem monitoring of carbon exchange from two oil palm plantations on converted tropical peat swamp forest. Our sites compare a newly converted oil palm plantation (OPnew) to a mature oil palm plantation (OPmature) and combine them in the context of existing emission factors. Mean annual net emission (NEE) of CO2 measured at OPnew during the conversion period (137.8 Mg CO2 ha‐1 yr ‐1) were an order of magnitude lower during the measurement period at OPmature (17.5 Mg CO2 ha‐1 yr‐1). However, mean water table depth (WTD) was shallower (0.26 m) than a typical drainage target of 0.6 m suggesting our emissions may be a conservative estimate for mature plantations, mean WTD at OPnew was more typical at 0.54 m. Reductions in net emissions were primarily driven by increasing biomass accumulation into highly productive palms. Further analysis suggested annual peat carbon losses of 24.9 Mg CO2‐C ha‐1 yr‐1 over the first 6 years, lower than previous estimates for this early period from subsidence studies, losses reduced to 12.8 Mg CO2‐C ha‐1 yr‐1 in the later, mature phase. Despite reductions in NEE and carbon loss over time, the system remained a large net source of carbon to the atmosphere after 12 years with the remaining 8 years of a typical plantation’s rotation unlikely to recoup losses. These results emphasise the need for effective protection of tropical peatlands globally and strengthening of legislative enforcement where moratoria on peatland conversion already exist. Publisher PDF

Agroforestry is a combination of monoculture agriculture and trees. Studies of net ecosystem exchange of CO2 (NEE) of agroforestry systems are rare, in comparison to the extensive studies of NEE of agricultural systems (croplands and grasslands). Agroforestry has been shown to alter the microclimate, productivity, and nutrient and water usage – as compared to standard agricultural practise. The, potentially, higher carbon sequestration of agroforestry, relative to monoculture systems, provides an interesting option for mitigating climate change, highlighting the need for improved study of agroforestry systems. The current study, as part of the SIGNAL (sustainable intensification of agriculture through agroforestry) project, investigates NEE of agroforestry compared to that of monoculture agriculture. The study employs paired comparisons of flux measurements above agroforestry and monoculture agronomy, replicated at five locations across Germany. Fluxes are measured, using innovative low-cost CO2 eddy covariance sensors (slow response Vaisala GMP343 IRGA with custom made housing), which have been successfully used in a study over grassland. Continuous data series from mid-summer until winter 2019 show that both systems acted as a sink with comparable fluxes during summer. The diurnal CO2 cycle and the response to management activities are distinguishable and in autumn preliminary results suggest a small difference in fluxes between the two systems. The low-cost eddy covariance system is able to capture the turbulence and to measure the CO2 flux over the agroforestry and monoculture agricultural system. We aim to further improve the quality of the CO2 fluxes, by adapting post-processing software to better estimate the difference in carbon uptake between the agroforestry and monoculture systems.

The recent expansion of oil palm (OP, Elaeis guineensis) plantations into tropical forest peatlands has resulted in ecosystem carbon emissions. However, estimates of net carbon flux from biomass changes require accurate estimates of the above ground biomass (AGB) accumulation rate of OP on peat. We quantify the AGB stocks of an OP plantation on drained peat in Malaysia from 3 to 12 years after planting using destructive harvests supported by non-destructive surveys of a further 902 palms. Peat specific allometric equations for palm (R2 = 0.92) and frond biomass are developed and contrasted to existing allometries for OP on mineral soils. Allometries are used to upscale AGB estimates to the plantation block-level. Aboveground biomass stocks on peat accumulated at ~6.39 ± 1.12 Mg ha−1 per year in the first 12 years after planting, increasing to ~7.99 ± 0.95 Mg ha−1 yr−1 when a ‘perfect’ plantation was modelled. High inter-palm and inter-block AGB variability was observed in mature classes as a result of variations in palm leaning and mortality. Validation of the allometries defined and expansion of non-destructive inventories across alternative plantations and age classes on peat would further strengthen our understanding of peat OP AGB accumulation rates.

The rheological characterization of human blood, through modeling and analysis of steady state, transient, and oscillatory shear flows, has made tremendous progress recently. Due to the aggregation of red blood cells at low shear rates, many recent models for blood rheology include a structural, thixotropic component with one of the most recent attempts unifying this approach with a viscoelastic formulation. We will show how these models, along with proposed modifications to another recent structural, kinetic thixotropy model, can improve modeling predictions. Results are compared to the Maxwell-like Bautista-Manero-Puig model, the Oldroyd-8 inspired viscoelastic Anand-Kwack-Masud model, a viscoelastic-thixotropic model from Blackwell and Ewoldt, and the Herschel-Bulkley model. We explore the weaknesses of the legacy blood models and then demonstrate the efficacy of the newly improved models for modeling human blood steady state and transient shear rheology to predict oscillatory shear flow.

Hydrogen embrittlement of notched martensitic advanced high-strength steels was studied. Notched specimens had (i) higher tensile strength and lower ductility, and (ii) increased hydrogen sensitivity, manifested by reductions in ductility and strength. Hydrogen-induced fast fractures (HIFF) initiated when the load-controlled specimen became mechanically unstable. The HIFF velocity (61−130 m/s) was greater than the velocity of ductile fracture (46 m/s). HIFF susceptibility increased with increasing steel strength, increasing hydrogen fugacity, and decreasing stress rate. HIFF exhibited brittle features (quasi-cleavage, transgranular and intergranular fracture) in the initiation zone, suggesting hydrogen-enhanced plasticity-mediated decohesion (HEPD). These features increased in area with increasing hydrogen embrittlement.

Glucagon-like peptide (GLP-1) is an endogenous hormone that induces insulin secretion from pancreatic islets and modified forms are used to treat diabetes mellitus type 2. Understanding how GLP-1 interacts with its receptor (GLP-1R) can potentially lead to more effective drugs. Modeling and NMR studies of the N-terminus of GLP-1 suggest a β-turn between residues Glu9-Phe12 and a kinked alpha helix between Val16-Gly37. N-terminal turn constraints attenuated binding affinity and activity (compounds 1-8). Lys-Asp (i, i+4) crosslinks in the middle and at the C-terminus increased alpha helicity and cAMP stimulation without much effect on binding affinity or beta-arrestin 2 recruitment (compounds 9-18). Strategic positioning of helix-inducing constraints and amino acid substitutions (Tyr16, Ala22) increased peptide helicity and produced ten-fold higher cAMP potency (compounds 19-28) over GLP-1(7-37)-NH2. The most potent cAMP activator (compound 23) was also the most potent inducer of insulin secretion.

Rule-of-five parameters and membrane permeabilities have been routinely used to guide development of orally bioavailabile drugs. Here we compare enantiomeric pairs of cyclic hexapeptides with identical rule-of-five parameters and membrane permeabilities. For each enantiomeric pair, the isomer with more l- than d-amino acids is much more orally bioavailable in rats, more metabolically stable to rat liver microsomes, and cleared more slowly in vivo.

Twisting or turning peptides: ring size and chi angle in side chain cross-linked tetrapeptides together control α- or β-turn structures, which mimic irregular secondary structures in proteins., Protein–protein interactions involve hotspots as small as 4 sequential amino acids. Corresponding tetrapeptides have no structure in water. Here we report linking side chains of amino acids X and Z to form 24 cyclic tetrapeptides, cyclo-[XAAZ]-NH2, and stabilise 14–18 membered rings that mimic different kinds of non-regular secondary structures found in protein hotspots. 2D NMR spectra allowed determination of 3D structures for 14 cyclic tetrapeptides in water. Five formed two (i, i + 3) hydrogen bonds and a beta/gamma (6, 7) or beta (9, 19, 20) turn; eight formed one (i, i + 4) hydrogen bond and twisted into a non-helical (13, 18, 21, 22, 24) or helical (5, 17, 23) alpha turn; one was less structured (15). A beta or gamma turn was favoured for Z = Dab, Orn or Glu due to a χ1 gauche (+) rotamer, while an alpha turn was favoured for Z = Dap (but not X = Dap) due to a gauche (–) rotamer. Surprisingly, an unstructured peptide ARLARLARL could be twisted into a helix when either a helical or non-helical alpha turn (5, 13, 17, 18, 21–24) with Z = Dap was attached to the N-terminus. These structural models provide insights into stability for different turns and twists corresponding to non-regular folds in protein hotspots.

Cyclization is an important post-translational modification of peptides and proteins that confers key advantages such as protection from proteolytic degradation, altered solubility, membrane permeability, bioavailability, and especially restricted conformational freedom in water that allows the peptide backbone to adopt the major secondary structure elements found in proteins. Non-ribosomal synthesis in bacteria, fungi, and plants or synthetic chemistry can introduce unnatural amino acids and non-peptidic constraints that modify peptide backbones and side chains to fine-tune cyclic peptide structure. Structures can be potentially altered further upon binding to a protein in biological environments. Here we analyze three-dimensional crystal structures for 211 bioactive cyclic peptides bound to 65 different proteins. The protein-bound cyclic peptides were examined for similarities and differences in bonding modes, for main-chain and side-chain structure, and for the importance of polarity, hydrogen bonds, hydrophobic effects, and water molecules in interactions with proteins. Many protein-bound cyclic peptides show backbone structures like those (strands, sheets, turns, helices, loops, or distorted variations) found at protein-protein binding interfaces. However, the notion of macrocycles simply as privileged scaffolds that primarily project side-chain substituents for complementary interactions with proteins is dispelled here. Unlike small-molecule drugs, the cyclic peptides do not rely mainly upon hydrophobic and van der Waals interactions for protein binding; they also use their main chain and side chains to form polar contacts and hydrogen bonds with proteins. Compared to small-molecule ligands, cyclic peptides can bind across larger, polar, and water-exposed protein surface areas, making many more contacts that can increase affinity, selectivity, biological activity, and ligand-receptor residence time. Cyclic peptides have a greater capacity than small-molecule drugs to modulate protein-protein interfaces that involve large, shallow, dynamic, polar, and water-exposed protein surfaces.

Wollamides are cyclic hexapeptides, recently isolated from an Australian soil Streptomyces isolate, that exhibit promising in vitro antimycobacterial activity against Mycobacterium bovis Bacille Calmette Guérin without displaying cytotoxicity against a panel of mammalian cells. Here, we report the synthesis and antimycobacterial activity of 36 new synthetic wollamides, collated with all known synthetic and natural wollamides, to reveal structure characteristics responsible for in vitro growth-inhibitory activity against Mycobacterium tuberculosis (H37Rv, H37Ra, CDC1551, HN878, and HN353). The most potent antimycobacterial wollamides were those where residue VI d-Orn (wollamide B) was replaced by d-Arg (wollamide B1) or d-Lys (wollamide B2), with all activity being lost when residue VI was replaced by Gly, l-Arg, or l-Lys (wollamide B3). Substitution of other amino acid residues mainly reduced or ablated antimycobacterial activity. Significantly, whereas wollamide B2 was the most potent in restricting M. tuberculosis in vitro, wollamide B1 restricted M. tuberculosis intracellular burden in infected macrophages. Wollamide B1 synergized with pretomanid (PA-824) in inhibiting M. tuberculosis in vitro growth but did not antagonize prominent first- and second-line tuberculosis antibiotics. Furthermore, wollamide B1 exerted bactericidal activity against nonreplicating M. tuberculosis and impaired growth of multidrug- and extensively drug-resistant clinical isolates. In vivo pharmacokinetic profiles for wollamide B1 in rats and mice encourage further optimization of the wollamide pharmacophore for in vivo bioavailability. Collectively, these observations highlight the potential of the wollamide antimycobacterial pharmacophore.

The vast size of oil palm (Elaeis guineensis) plantations has led to lightweight unmanned aerial vehicles (UAVs) being identified as cost effective tools to generate inventories for improved planta...

Estimates of peatland carbon fluxes based on remote sensing data are a useful addition to monitoring methods in these remote and precious ecosystems, but there are questions as to whether large-scale estimates are reliable given the small-scale heterogeneity of many peatlands. Our objective was to consider the reliability of models based on Earth Observations for estimating ecosystem photosynthesis at different scales using the Forsinard Flows RSPB reserve in Northern Scotland as our study site. Three sites across the reserve were monitored during the growing season of 2017. One site is near-natural blanket bog, and the other two are at different stages of the restoration process after removal of commercial conifer forestry. At each site we measured small (flux chamber) and landscape scale (eddy covariance) CO2 fluxes, small scale spectral data using a handheld spectrometer, and obtained corresponding satellite data from MODIS. The variables influencing GPP at small scale, including microforms and dominant vegetation species, were assessed using exploratory factor analysis. A GPP model using land surface temperature and a measure of greenness from remote sensing data was tested and compared to chamber and eddy covariance CO2 fluxes; this model returned good results at all scales (Pearson's correlations of 0.57 to 0.71 at small scale, 0.76 to 0.86 at large scale). We found that the effect of microtopography on GPP fluxes at the study sites was spatially and temporally inconsistent, although connected to water content and vegetation species. The GPP fluxes measured using EC were larger than those using chambers at all sites, and the reliability of the TG model at different scales was dependent on the measurement methods used for calibration and validation. This suggests that GPP measurements from remote sensing are robust at all scales, but that the methods used for calibration and validation will impact accuracy.

This study examines the efficacy of blended learning – an approach that combines in-class and online methods in a way that leverages the strengths of both – using a field experiment spanning 16 weeks. An information-processing model of learning suggests that learners will weigh the cost of information accessibility against its value in ways that will impact their interactions with the available information sources, which will consequently affect learning outcomes. Results of our study suggest that such an assessment did indeed occur and that it impacted learning performance. Specifically, our results support the idea that providing high-value content in both settings – the classroom rich, yet high cost and online efficient, yet low cost, enhances performance. The largest gains in performance were seen by those who used the blended learning system the most, with the lowest gains by those who did not use the system at all i.e. the control group.

Cyclic pentapeptides (e.g. Ac-(cyclo-1,5)-[KAXAD]-NH2; X = Ala, 1; Arg, 2) in water adopt one alpha-helical turn defined by three hydrogen bonds. NMR structure analysis reveals a slight distortion from alpha-helicity at the C-terminal aspartate caused by torsional restraints imposed by the K(i)-D(i + 4) lactam bridge. To investigate this effect on helix nucleation, the more water-soluble 2 was appended to N-, C-, or both termini of a palindromic peptide ARAARAARA (

Natural killer T (NKT) cells that express the semi-invariant T-cell receptor are innate-like lymphocytes whose functions are controlled by self and foreign glycolipid ligands. Such ligands are presented by the antigen-presenting, MHC class I-like molecule CD1d, which belongs to a family of lipid antigen-presenting molecules collectively called CD1. Activation of NKT cells in vivo results in rapid release of copious amounts of effector cytokines and chemokines with which they regulate innate and adaptive immune responses to pathogens, certain types of cancers and self-antigens. The nature of CD1d-restricted ligands, the manners in which they are recognised and the unique effector functions of NKT cells suggest an immunoregulatory role for this T-cell subset. Their ability to respond fast and our ability to steer NKT cell cytokine responses to altered lipid ligands make them an important target for vaccine design and immunotherapies against autoimmune diseases. This article summarises our current understanding of CD1d-restricted NKT cell biology and how these innate-like lymphocytes control inflammation. Key Concepts CD1d molecules belong to a family of lipid antigen-presenting molecules collectively called CD1. CD1d molecules resemble peptide antigen-presenting MHC class I molecules. CD1d molecules restrict the specificity and functions of Natural killer T (NKT) cells. NKT cells recognise self- and nonself-lipid ligands. Antigen recognition quickly activates NKT cells, which respond immediately by releasing proinflammatory and immunoregulatory cytokines and chemokines. Activated NKT cells communicate with cells of the innate and adaptive immune systems by cell–cell interactions and/or via soluble mediators. Such communications allow NKT cells to control immune responses to microbial pathogens and certain cancers. NKT-cell activation has beneficial and, sometimes, adverse effects on certain autoimmune and metabolic disorders. NKT cells are targets for vaccine adjuvants and immunotherapies. Keywords: CD1d molecules; NKT cells; glycolipid-reactive T cells; innate-like T cells; glycosphingolipids; α-galactosylceramide; immunoregulation

Consumer demand for Internet of Things (IoT) devices is projected to continue its lucrative acceleration despite growing cybersecurity concerns and incidents reported in the public press. We explore this apparent anomaly through a model, based on the relevant literature, casting consumers’ IoT Purchase Intention as driven by their pre-existing IoT Risk Knowledge, their level of IoT-related Security Concern, and their device-specific perceptions of Riskiness and Coolness. Based on this model, we collected survey data gauging Purchase Intention for four products varying in Riskiness and Coolness levels. Our analysis extends prior research by confirming the negative influence of IoT Risk Knowledge, implicating it as an emerging drag on IoT consumer demand growth. We further confirm the expected effects of Device Riskiness (negative) and Coolness (positive) as primary factors and we note the cognitive dissonance implied by the inherent IoT device trade-off: greater Coolness tends to come at the price of greater Riskiness. In this trade-off, Coolness mattered more for consumers with lower IoT Security Concern suggesting greater susceptibility to the “wow” factor as offsetting or even distracting from the associated risk. This work contributes to the literature by confirming antecedents and revealing subtleties in the interplay of general and device-specific factors affecting consumer IoT Purchase Intention, shedding light on the feature-attraction/risk-avoidance paradox and identifying implications for both research and practice as the marketplace and consumer perceptions co-evolve going forward.

The cyclic heptapeptide derivative, sanguinamide A, is a model scaffold for studying how component amino acids, heterocycles, and N-methylation influence membrane permeability and oral bioavailability. Membrane permeable sanguinamide A analogues have been reported, but there is limited data on their pharmacokinetic properties invivo. Here we report pharmacokinetic properties for highly cell and membrane permeable sanguinamide A analogues in rats and find that there is no correlation between reported permeability invitro and oral bioavailability invivo. We show that N-methylation of sanguinamide A analogues gives compounds with greater flexibility, greater susceptibility to degradation by rat liver microsomes, and lower oral bioavailability in rats.

Correction for ‘Twists or turns: stabilising alpha vs. beta turns in tetrapeptides’ by Huy N. Hoang et al., Chem. Sci., 2019, 10, 10595–10600, DOI: 10.1039/C9SC04153B.

Peatlands are an important terrestrial carbon store, but disturbance has resulted in the degradation of many peatland ecosystems and caused them to act as a net carbon source. Restoration work is being undertaken but monitoring the success of these schemes can be difficult and costly using traditional field-based methods. A landscape-scale alternative is to use satellite data to assess the condition of peatlands and to estimate gaseous carbon fluxes. In this study we used Moderate Resolution Imaging Spectroradiometer (MODIS) products to model Gross Primary Productivity (GPP) over peatland sites at various stages of restoration. We found that the MOD17A2H GPP product overestimates GPP modelled from data collected by eddy covariance towers situated at two ex-forestry sites undergoing restoration towards blanket bog at the Forsinard Flows RSPB reserve, Scotland, UK (one full year of data), and a near-natural Atlantic blanket bog site in Glencar, Ireland (ten-year data series). We calibrated a Temperature and Greenness (TG) model for the Forsinard sites and found it to be more accurate than the MODIS GPP product at local scale. We also found that inclusion of a wetness factor using the Normalised Difference Water Index (NDWI) improved inter-annual accuracy of the model. This TGWa (annual Temperature, Greenness and Wetness) model was then applied to six control sites comprising near-natural bog across the reserve, and to six sites on which restoration began between 1998 and 2006. GPP from 2005 to 2016 was estimated for each site using the model. The resulting modelled trends are positive at all six restored sites, increasing by approximately 5.5 g C/m2/yr every year since restoration began in the Forsinard Flows reserve. The results suggest that peatland sites undergoing restoration at Forsinard Flows reach the carbon assimilation potential of near-natural bog sites between 5 and 10 years after restoration was begun.

This paper presents details of the indoor wideband and directional propagation measurements at 26 GHz in which a wideband channel sounder using a millimeter wave (mmWave) signal analyzer and vector signal generator was employed. The setup provided 2 GHz bandwidth and the mechanically steerable directional lens antenna with 5 degrees beamwidth provides 5 degrees of directional resolution over the azimuth. Measurements provide path loss, delay and spatial spread of the channel. Angular and delay dispersion are presented for line-of-sight (LoS) and non-line-of-sight (NLoS) scenarios.

Most protein-protein interactions occur inside cells. Peptides can inhibit protein-protein interactions but tend not to enter cells. We systematically compare cell permeability for 8-12 residue model peptides with helix-inducing lactam/hydrocarbon linkers between amino acid sidechains. Cell uptake increases when hydrophobic residues and lactam linkers (i, i + 4) form a contiguous hydrophobic surface patch. Uptake increases further when both hydrophobic and positively charged (but not neutral or negative) residues are clustered into like surface patches. Amphipathicity alone is however insufficient for cell uptake of acyclic sequences. Changing the linker from lactam to hydrocarbon further increases uptake, but also promotes cell lysis. Helicity, positive charge and amphipathicity together promote cell permeability. Most known bioactive helical peptides do not optimally cluster residues for amphipathicity and so are likely unoptimised for cell uptake.

Cyclic peptides and macrocycles have the potential to be membrane permeable and orally bioavailable, despite often not complying with the "rule of five" used in medicinal chemistry to guide the discovery of oral drugs. Here we compare solvent-dependent three-dimensional structures of three cyclic hexapeptides containing d-amino acids, prolines, and intramolecular hydrogen bonds. Conformational rigidity rather than flexibility resulted in higher membrane permeability, metabolic stability and oral bioavailability, consistent with less polar surface exposure to solvent and a reduced entropy penalty for transition between polar and nonpolar environments.

Once a burgeoning field of study, over the past decade or so, T cell epitope discovery has lost some luster. The contributory factors perchance are the general notion that any newly discovered epitope will reveal very little about an immune response and that knowledge of epitopes are less critical for vaccine design. Despite these notions, the breadth and depth of T cell epitopes derived from clinically important microbial agents of human diseases largely remain ill defined. We review here a flurry of recent reports that have rebirthed the field. These reports reveal that epitope discovery is an essential step toward rational vaccine design and critical for monitoring vaccination efficacy. The new findings also indicate that neither immunogenicity nor immunodominance predict protective immunity. Hence, an immunogenic epitope is but a peptide unless proven protective against disease.

Pollsters expected a hung parliament, but UK voters instead returned a small Conservative majority. Timothy Martyn Hill reviews the predictions – and the errors – that were made

Cyclic constraints are incorporated into an 11-residue analogue of the N-terminus of glucagon-like peptide-1 (GLP-1) to investigate effects of structure on agonist activity. Cyclization through linking side chains of residues 2 and 5 or 5 and 9 produced agonists at nM concentrations in a cAMP assay. 2D NMR and CD spectra revealed an N-terminal β-turn and a C-terminal helix that differentially influenced affinity and agonist potency. These structures can inform development of small molecule agonists of the GLP-1 receptor to treat type 2 diabetes.

Protease activated receptor 2 (PAR2) is associated with metabolism, obesity, inflammatory, respiratory and gastrointestinal disorders, pain, cancer, and other diseases. The extracellular N-terminus of PAR2 is a common target for multiple proteases, which cleave it at different sites to generate different N-termini that activate different PAR2-mediated intracellular signaling pathways. There are no synthetic PAR2 ligands that reproduce the same signaling profiles and potencies as proteases. Structure–activity relationships here for 26 compounds spanned a signaling bias over 3 log units, culminating in three small ligands as biased agonist tools for interrogating PAR2 functions. DF253 (2f-LAAAAI-NH2) triggered PAR2-mediated calcium release (EC50 2 μM) but not ERK1/2 phosphorylation (EC50 > 100 μM) in CHO cells transfected with hPAR2. AY77 (Isox-Cha-Chg-NH2) was a more potent calcium-biased agonist (EC50 40 nM, Ca2+; EC50 2 μM, ERK1/2), while its analogue AY254 (Isox-Cha-Chg-A-R-NH2) was an ERK-biased agonis...

Today, many firms are striving to enhance customers’ purchase intentions by leveraging online consumer communities but a better understanding of the underlying phenomena is needed to guide their efforts most effectively. In this study we analyse how three hotel firms engage potential customers through FlyerTalk, an online consumer community of frequent travellers (FlyerTalk.com). Our results suggest that for some firms, the frequency and perceived value of their participation are two important factors that can potentially be managed toward positive purchasing intention effects.

Many proteins exert their biological activities through small exposed surface regions called epitopes that are folded peptides of well-defined three-dimensional structures. Short synthetic peptide sequences corresponding to these bioactive protein surfaces do not form thermodynamically stable protein-like structures in water. However, short peptides can be induced to fold into protein-like bioactive conformations (strands, helices, turns) by cyclization, in conjunction with the use of other molecular constraints, that helps to fine-tune three-dimensional structure. Such constrained cyclic peptides can have protein-like biological activities and potencies, enabling their uses as biological probes and leads to therapeutics, diagnostics and vaccines. This Review highlights examples of cyclic peptides that mimic three-dimensional structures of strand, turn or helical segments of peptides and proteins, and identifies some additional restraints incorporated into natural product cyclic peptides and synthetic macrocyclic peptidomimetics that refine peptide structure and confer biological properties.

Viele Proteine entfalten ihre biologische Aktivitat uber kleine exponierte Oberflachenregionen aus gefalteten Peptiden mit wohldefinierten dreidimensionalen Strukturen, Epitope genannt. Kurze synthetische Peptidsequenzen, die diesen bioaktiven Proteinoberflachen entsprechen, bilden in Wasser keine thermodynamisch stabilen proteinahnlichen Strukturen. Die Bildung proteinahnlicher biologisch aktiver Konformationen (Strange, Helices, Kehren) kann jedoch durch Cyclisierung in Verbindung mit anderen molekularen Verstrebungen induziert werden. Letztere helfen bei der Feinabstimmung der dreidimensionalen Struktur. Solche fixierten cyclischen Peptide konnen ahnliche biologische Aktivitaten und Wirkungen wie das als Vorbild dienende Protein haben, sodass sie als biologische Sonden und Leitstrukturen fur Therapeutika, Diagnostika und Impfstoffe dienen konnen. Dieser Aufsatz beleuchtet Beispiele fur cyclische Peptide, die dreidimensionale Strukturen von Strang-, Kehren- oder Helixabschnitten von Peptiden und Proteinen nachahmen und beschreibt einige weitere Elemente, die in cyclisch peptidischen Naturstoffen und synthetischen makrocyclischen Peptidomimetika vorkommen, dort die Peptidstruktur verfeinern und ihr biologische Aktivitaten verleihen.

HIV-1 Rev is a 116 residue transporter protein that enters the host cell nucleus and uses its 17 amino acid segment (Rev34–50) to bind and capture a specific piece of RNA, the Rev Response Element (RRE), for transport to the cytoplasm. This is critical for HIV replication. In isolation, Rev34–50 shows negligible structure in water, but is alpha helical in a mixture of water and 2,2,2-trifluoroethanol (TFE) or when bound to RRE. Here we report that helix-constrained cyclic pentapeptides, either appended to the N-terminus or incorporated within Rev34–50, are efficient helix nucleators in water. They induce up to 90% alpha helicity for isolated Rev peptides in water and confer high RNA-binding affinity.

The use of peptides in medicine is limited by low membrane permeability, metabolic instability, high clearance, and negligible oral bioavailability. The prediction of oral bioavailability of drugs relies on physicochemical properties that favor passive permeability and oxidative metabolic stability, but these may not be useful for peptides. Here we investigate effects of heterocyclic constraints, intramolecular hydrogen bonds, and side chains on the oral bioavailability of cyclic heptapeptides. NMR-derived structures, amide H-D exchange rates, and temperature-dependent chemical shifts showed that the combination of rigidification, stronger hydrogen bonds, and solvent shielding by branched side chains enhances the oral bioavailability of cyclic heptapeptides in rats without the need for N-methylation.

Development of peptide-based drugs has been severely limited by lack of oral bioavailability with less than a handful of peptides being truly orally bioavailable, mainly cyclic peptides with N-methyl amino acids and few hydrogen bond donors. Here we report that cyclic penta- and hexa-leucine peptides, with no N-methylation and five or six amide NH protons, exhibit some degree of oral bioavailability (4-17%) approaching that of the heavily N-methylated drug cyclosporine (22%) under the same conditions. These simple cyclic peptides demonstrate that oral bioavailability is achievable for peptides that fall outside of rule-of-five guidelines without the need for N-methylation or modified amino acids.

Helix-constrained polypeptides have attracted great interest for modulating protein-protein interactions (PPI). It is not known which are the most effective helix-inducing strategies for designing PPI agonists/antagonists. Cyclization linkers (X1-X5) were compared here, using circular dichroism and 2D NMR spectroscopy, for α-helix induction in simple model pentapeptides, Ac-cyclo(1,5)-[X1-Ala-Ala-Ala-X5]-NH2, in water. In this very stringent test of helix induction, a Lys1→Asp5 lactam linker conferred greatest α-helicity, hydrocarbon and triazole linkers induced a mix of α- and 3₁₀-helicity, while thio- and dithioether linkers produced less helicity. The lactam-linked cyclic pentapeptide was also the most effective α-helix nucleator attached to a 13-residue model peptide.