Rational Platform Research Articles

Solar Fuel Synthesis Using a Semiartificial Colloidal Z-Scheme.

The integration of enzymes with semiconductor light absorbers in semiartificial photosynthetic assemblies offers an emerging strategy for solar fuel production. However, such colloidal biohybrid systems rely currently on sacrificial reagents, and semiconductor-enzyme powder systems that couple fuel production to water oxidation are therefore needed to mimic an overall photosynthetic reaction. Here, we present a Z-scheme colloidal enzyme system that produces fuel with electrons sourced from water. This "closed-cycle" semiartificial approach utilizes particulate SrTiO3:La,Rh and BiVO4:Mo (light absorbers), hydrogenase or formate dehydrogenase (cocatalyst), and a molecular cobalt complex (a redox mediator). Under simulated solar irradiation, this system continuously generates molecular hydrogen or formate, while co-producing molecular oxygen for 10 h using only sunlight, water, and carbon dioxide as inputs. In-depth analysis using quartz crystal microbalance, photoelectrochemical impedance spectroscopy, transient photocurrent spectroscopy, and intensity-modulated photovoltage spectroscopy provides mechanistic understanding and characterization of the semiconductor-enzyme hybrid interface. This study provides a rational platform to assemble functional semiartificial colloidal Z-scheme systems for solar fuel synthesis.

Ethylene glycol derivative-pendanted quaternary ammonium antibacterial polymers with low cytotoxicity

Cationic poly(quaternary ammonium) (pQA) polymers exhibit excellent antibacterial efficacy but high cytotoxicity (thus low selectivity), hampering their widespread use as antibacterial agents. Herein, we systematically synthesized a series of pQA polymers (pQA−EG) containing biocompatible ethylene glycol (EG) derivative pendant groups with different chain lengths and connectivities (acyclic/cyclic). The antibacterial efficacy of pQA−EG was optimized by a moderate EG chain length, which endowed the polymer with balanced hydrophobicity/charge density. The high chain flexibility of the EG derivative combined with its good hydrophilicity was critical for significantly lowering the cytotoxicity of pQA−EG. Hence, compared with pQA polymers containing alkyl pendant groups, the pQA−EG polymer containing a long flexible EG derivative pendant group exhibited significantly lower cytotoxicity and higher selectivity as well as satisfactory antibacterial activity and kinetics. Our study offers a rational platform for synthesizing low-toxicity antibacterial polymers and provides fundamental insights into their structure−biological activity relationships.

Optimizing Immunoglobulin Replacement Therapy for Patients with B-Cell Malignancies and Hypogammaglobulinemia: The Investigator-Initiated, International, Randomized Phase II/III Rational Platform Trial, and the Rationalise (STOP IgRT) Domain

Background & significance: Hypogammaglobulinemia is common in patients with chronic lymphocytic leukemia (CLL), non-Hodgkin lymphoma (NHL) and multiple myeloma (MM), and associated with an increased risk of serious infection. Immunoglobulin (Ig) replacement therapy (IgRT) is widely used to reduce this risk, but national guidelines and clinical practice are variable. Supportive care and treatment regimens have evolved since evidence supporting IgRT was published. Ig is expensive, in limited supply, and with increasing use of B-cell-depleting therapies in this setting, and prolonged survival, demand continues to grow globally.It is not clear when to start IgRT, nor the optimal duration or dose. Some jurisdictions require a trial of oral antibiotics (Abx) prior to starting IgRT, and some recommend a trial of stopping IgRT in stable patients. More data are needed on the benefits, risks and costs of these different approaches. Hypothesis: For the prevention of infection in adults with hypogammaglobulinemia due to CLL, NHL or MM prophylactic oral Abx, or a strategy of patient-held emergency Abx supply only, are non-inferior to IgRT. Study design: The RATIONAL platform trial is an investigator-initiated, international (Australia, New Zealand, Canada), phase II/III randomized controlled trial (RCT) with 3 domains: (1) the RATIONALISE ‘STOP Ig’ study (ACTRN 12622000359730), now recruiting; (2) a ‘START Ig’ study comparing IgRT or Abx in patients eligible to start IgRT; and (3) a DOSE study comparing IgRT at 0.25 vs 0.4g/kg/month (m). The platform includes qualitative, biological (immune profile, microbiome) and health economics studies. In the RATIONALISE (‘STOP Ig‘) domain, patients are randomized 1:1:1 to either: - Arm A: Stop Ig and commence prophylactic oral Abx: Daily trimethoprim-sulfamethoxazole (co-trimoxazole) 160mg/800mg (doxycycline 100mg/day if co-trimoxazole hypersensitive). - Arm B: Stop Ig. Keep amoxycillin/clavulanic acid 1750-2000mg/250mg and ciprofloxacin 750mg on hand for initial use for symptoms of infection, with immediate clinical review. Clindamycin 600mg if penicillin hypersensitive. - Arm C: Continue Ig: IV q 4±1 wks at 0.4g/kg, modified to achieve IgG trough of at least lower limit of age-specific serum IgG reference range, OR subcutaneous (SC), dosed in line with site/country criteria. Inclusion criteria: ≥18y with CLL, NHL or MM, receiving IgRT (IV or SC) for >6m, eligible for Ig cessation in opinion of treating clinician, life expectancy >12m, able to give informed consent and comply with each of the trial arms. Exclusion criteria: Prior/planned allogeneic transplantation. Major infection (≥Grade 3 CTCAE v5) in preceding 3m, and/or current active infection requiring antimicrobial treatment. Already on daily Abx for purpose of preventing bacterial infection (may have antiviral, antifungal, PJP prophylaxis). Intolerance of all trial Abx options.Communication, compliance or logistics likely to limit ability to take prophylactic or emergency Abx, or obtain urgent medical attention for infection symptoms. Pregnant/breastfeeding. Creatinine clearance <30 mL/min. Splenectomy. Enrolment deemed not in patient's interest. Primary endpoint: Event-free survival, from randomization until ≥Grade 3 infection or death from any cause. Secondary endpointsinclude other infectious outcomes to 12m, time free from hospitalization and therapeutic Abx, Abx resistance, QoL, trough Ig levels, proportion of patients who stop Ig and re-start, costs, adverse events. End-point determination: Study is not blinded, but infections and adverse events are adjudicated by an independent, blinded outcome adjudication committee. Planned treatment duration in all arms is 12m, with follow-up via established clinical registries. Statistical methods: STOP Ig domain has an initial cap of 300 patients and uses flexible Bayesian monitoring involving frequent interim analyses where interventions may be stopped early to declare proof-of-concept/efficacy or inferiority, or accrual may expand. Stratified by diagnosis. Progress/Summary: This novel international platform trial will inform infection prevention strategies for the growing population of patients with CLL, NHL or MM living with hypogammaglobulinemia. The first domain ('STOP Ig') is now open at 4 sites, with the first patients recruited. To our knowledge, this is the first trial of 'stopping' Ig in this setting.

Multi-functional core-shell pomegranate peel amended alginate beads for phenol decontamination and bio-hydrogen production: Synthesis, characterization, and kinetics investigation

Phenolic compounds discharged from different industries comprise a great threatening to environmental system and human health. They are toxic even at a low concentration (<1.0 μg/L). Adsorption is attractive scenario for successful removal of phenolic compounds from wastewater. Herein, a facile strategy was employed to fabricate amended alginate beads based on pomegranate peel. The constructed beads were intensively scrutinized, and introduced for the remediation of phenol from wastewater under various operational parameters of pHi (e.g., 2.1–10.7), adsorbent dose (e.g., 0.5 – 5.0 g L−1), initial phenol concentrations (e.g., 10.0 – 500.0 mg L−1), and residence time (e.g., 5.0 – 270.0 min). Consequently, the efficacy of bio-hydrogen production via dark fermentative process of co-substrate metabolism of phenol and glucose in presence of beads was systematically inspected. Our findings revealed that the maximum adsorption capacity of beads was 119.048 mg g−1 at pH 7.2. Comfortingly, the fitting isotherms experiments expounded that the adsorption of phenol by beads conformed to the Langmuir model. The kinetics modelling findings matched with pseudo-second-order model. Otherwise, the as-prepared beads have remarkably improved anaerobic co-substrate fermentation process of glucose (synergism) and phenol removal efficiency (91.1–97.0%), with bio-hydrogen yield of 0.35 up to 2.65 mol-H2 mol−1 glucose. Overall, the present study illustrates an ample perspective of natural resources feasibility as a futuristic rational platform for wastewater decontamination and bio-hydrogen production.

A Spatiotemporal Bidirectional Attention-Based Ride-Hailing Demand Prediction Model: A Case Study in Beijing During COVID-19

The COVID-19 pandemic has severely affected urban transport patterns, including the way residents travel. It is of great significance to predict the demand of urban ride-hailing for residents’ healthy travel, rational platform operation, and traffic control during the epidemic period. In this paper, we propose a deep learning model, called MOS-BiAtten, based on multi-head spatial attention mechanism and bidirectional attention mechanism for ride-hailing demand prediction. The model follows the encoder-decoder framework with a multi-output strategy for multi-steps prediction. The pre-predicted result and the historical demand data are extracted as two aspects of bidirectional attention flow, so as to further explore the complicated spatiotemporal correlations between the historical, present and future information. The proposed model is evaluated on the real-world dataset during COVID-19 in Beijing, and the experimental results demonstrate that MOS-BiAtten achieves a better performance compared with the state-of-art methods. Meanwhile, another dataset is used to verify the generalization performance of the model.

Hinduism, Belief and the Colonial Invention of Religion: A before and after Comparison

As known from the academic literature on Hinduism, the foreign, Persian word, “Hindu” (meaning “Indian”), was used by the British to name everything indigenously South Asian, which was not Islam, as a religion. If we adopt explication as our research methodology, which consists in the application of the criterion of logical validity to organize various propositions of perspectives we encounter in research in terms of a disagreement, we discover: (a) what the British identified as “Hinduism” was not characterizable by a shared set of beliefs or shared outlook, but a disagreement or debate about basic topics of philosophy with a discourse on tenets of moral philosophy anchoring the debate; and (b), the Western tradition’s historical commitment to language as the vehicle of thought not only leads to the conflation of propositions with beliefs, but to interpreting (explaining by way of belief) on the basis of the Eurocentric tradition rooted exclusively in ancient Greek philosophy. Interpretation on the basis of the Western tradition leads to the Western tradition vindicating itself as the non-traditional, non-religious, rational platform—the secular—for explaining everything—the residua are what get called religions on a global scale. This serves the political function of insulating Western colonialism from indigenous moral and political criticism. Given that Western colonialism is the pivotal event, before which South Asians just had philosophy, and after which they had religion (the explanatory residua of Eurocentric interpretation), we can ask about Hindu religious belief. This only pertains to the period after colonialism, when Hindus adopted a Westcentric frame for understanding their tradition as religious because of colonization. Prior to this, the tradition the British identified as “Hindu” had a wide variety of philosophical approaches to justification, which often criticized propositional attitudes, like belief, as irrational.

Cinnamaldehyde and Curcumin Prime Akt2 for Insulin-Stimulated Activation.

In this study, the effects of cinnamaldehyde and curcumin on Akt2, a serine/threonine protein kinase central to the insulin signaling pathway, were examined in preadipocytes. Cinnamaldehyde or curcumin treatment increased Akt2 phosphorylation at multiple sites including T450 and Y475, but had no effect on Akt2 phosphorylation at S474, which is critical for Akt2 activation. Surprisingly, insulin treatment with cinnamaldehyde or curcumin increased p-Akt2 (S474) by 3.5-fold versus insulin treatment alone. Furthermore, combined cinnamaldehyde, curcumin, and insulin treatment increased p-Akt2 (S474) by 7-fold versus insulin treatment alone. Interestingly, cinnamaldehyde and curcumin inhibited both serine/threonine phosphatase 2A (PP2A) and protein tyrosine phosphatase 1B (PTP1B). Akt2 activation is a multistep process that requires phosphorylation at T450 for proper folding and maturation, and phosphorylation of both Y475 and S474 for stabilization of the catalytic domain. It is plausible that by inhibiting PP2A and PTP1B, cinnamaldehyde and curcumin increase phosphorylation at T450 and Y475, and prime Akt2 for insulin-stimulated phosphorylation at S474. Notably, the combination of a PP2A inhibitor, okadaic acid, and a PTP1B inhibitor increased p-Akt2 (S474), even in the absence of insulin. Future combinations of PP2A and PTP1B inhibitors provide a rational platform to engineer new therapeutics for insulin resistance syndrome.

Porous Nanofiber Membrane: Rational Platform for Highly Sensitive Thermochromic Sensor (Adv. Funct. Mater. 24/2022)

Porous Nanofiber Membranes In article number 2200463, Il-Doo Kim and co-workers develop highly sensitive thermochromic sensors using nanofiber (NF) membranes with tunable alignments of the NFs via a fine-tuned electric field during the electrospinning process. The porous NFs membrane-based sensors show highly sensitive thermochromic sensing performance at 31.6–42.7 °C. The developed NFs sensors membranes could be further integrated with masks, patches, and bracelets for wearable applications.

Porous Nanofiber Membrane: Rational Platform for Highly Sensitive Thermochromic Sensor

AbstractThermochromic sensors provide an intuitive and real‐time solution for monitoring the local temperature with naked eyes. Conventional thermochromic sensors often utilize either solution‐type or dense film‐type platforms, which are suboptimal morphologies for exposing a large number of dye molecules to the surface, leading to low sensitivity and sluggish responding speeds. Herein, this article introduces rational synthetic routes to fabricate highly sensitive nanofiber (NF) sensor membranes loaded with thermochromic dyes (C3H6N6·CH2O)x‐loaded nanofibers (NFs) sensor membranes by alignment‐controllable electrospinning techniques (x–y perpendicular and rotary). The NF‐based porous sensor membranes exhibit two‐ to fivefold improved thermochromic sensitivity (ΔRGB) compared to those of dense film‐type sensors at 31.6–42.7 °C. This is attributed to the uniform distribution of dyes throughout the porous NF structure (≈95.7%), which exhibits excellent light transmittance that is 10–30‐fold higher than that of film‐type sensors. Based on the available shape‐conforming synthetic strategies, this article further demonstrates wearable thermochromic sensors in the forms of mask‐, patch‐, and bracelet‐type devices, which can accurately monitor body temperature in real time.

Bio-separator and bio-synthesizer of metallic nanoparticles - A new vision in bioremediation

The conception of a bio-separator and bio-synthesizer synchronics of nanoparticles (NPs) from dead cells of the yeast Rhodotorula mucilaginosa (R. mucilaginosa), presents itself as a strong candidate for bioremediation. In this research, the dead yeast cells allowed the synthesis and simultaneous separation of silver, copper and nickel NPs, in aqueous medium, under green physical–chemical conditions. Through the yeast cell wall occurred, the formation of two bimetallic systems of NPs: one of Ni/NiO NPs extracellularly - Ag/Ag2O NPs intracellularly, and another of Ni/NiO NPs extracellularly - Cu NPs intracellularly. This sustainable protocol represents an efficient metals uptake method. Such natural bioprocess is a rational platform for the distinct synthesis of NPs (extra and intracellularly), for possible use in nanobioremediation of sites impacted by multiple toxic metals, being easy to handle, effective cost, and concomitant synthesis of NPs.

Genome-wide analysis of glutathione S-transferase gene family in chickpea suggests its role during seed development and abiotic stress.

Glutathione S-transferases (GSTs) are multifunctional proteins that help in oxidative stress metabolism and detoxification of xenobiotic compounds. Studies pertaining to GST gene family have been undertaken in various plant species, however no information is available with respect to GST genes in chickpea. In the current study, we identified a total of 51 GST encoding genes in chickpea (CaGST) genome. Phylogenetic analysis revealed that GST gene family can be divided into eleven distinct classes. Tau and phi were the major classes in chickpea and one third of the CaGST genes represented segmental duplication and purifying selection was common among these genes. Expression of many CaGST genes, in particular, members of tau class were found to be upregulated under abiotic stress conditions. In addition, CaGST genes displayed differential expression patterns across diverse organs/tissues, suggesting their roles in developmental processes. Many CaGST genes showed opposite expression pattern in small- and large-seeded chickpea cultivars during seed development. Higher expression of CaGST genes in small-seeded cultivar at maturation stages of seed development suggested their important role in seed development and seed size/weight determination in chickpea. Overall, these results provide a comprehensive information on GST gene family members in chickpea and is expected to provide a rational platform to explore versatile role of these genes in semi-arid legume crops.

Abstract 2657: GSTP mediated S-glutathionylation of GRP78 attributes to multiple myeloma resistance

Abstract The proteasome inhibitor bortezomib (Btz) is used in the treatment of multiple myeloma (MM), but its efficacy is restricted by the wide-spread occurrence of resistance. MM cells have the highest rate of secretion of proteins with disulfide bonds and are exquisitely dependent upon redox balance and accurate protein folding. Our results showed that some important redox enzymes, including glutathione transferase P (GSTP) are upregulated in Btz-resistant cells. While GSTP was originally described as a phase II detoxification enzyme, our laboratory has ascribed a thiolase activity catalyzing the forward S-glutathionylation reaction, a post-translational modification (PTM) through reversible addition of glutathione to thiolate anions of cysteines in target proteins. Although primarily cytosolic, GSTP has been reported in the nuclear and mitochondrial compartments. We recently showed that GSTP is localized at high concentrations in the endoplasmic reticulum (ER) and regulates a number of redox active proteins through S-glutathionylation; these include glucose-regulated protein (GRP78). Proteomic analysis revealed that both cys41 and cys 420 of GRP78 were subject to S-glutathionylation and this PTM altered the activity of GRP78, enhancing its holdase activity, while concomitantly decreasing its foldase activity. This alters protein folding and contributes to the Btz-resistant phenotype of MM. Our data also support the principle that drugs that target the ER protein folding machinery significantly alter local redox conditions. Thapsigargin (Thg) is an inhibitor of SERCA, interfering with the functional activities of calcium-dependent chaperones, while tunicamycin (TuM) inhibits N-linked glycosylation. Btz resistant cells showed marked levels of cross-resistance (2-10 fold) to both. Linking such results with GSTP/redox, the absence of GSTP (from GSTP knockout mice) makes both bone marrow derived dendritic and mouse embryonic fibroblast cells more sensitive to Btz, ThG and TuM. The nature of the cross-resistance patterns and the fact that the presence of GSTP in drug naïve cells also confers resistance provides a rational platform for studying redox-based mechanisms as causative in resistance. Moreover, inhibition of GSTP with Telintra restored Btz sensitivity, decreased S-glutathionylation and increased expression of markers of UPR induced apoptosis. We conclude that altered GSTP expression and S-glutathionylation of key ER-localized proteins are significant contributors to drug response and resistance in MM treated with Btz. Citation Format: Zhiwei Ye, Jie Zhang, Kenneth D. Tew, Danyelle M. Townsend. GSTP mediated S-glutathionylation of GRP78 attributes to multiple myeloma resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2657.

A multi-method and structure-based in silico vaccine designing against Echinococcus granulosus through investigating enolase protein.

Introduction: Hydatid disease is a ubiquitous parasitic zoonotic disease, which causes different medical, economic and serious public health problems in some parts of the world. The causal organism is a multi-stage parasite named Echinococcus granulosus whose life cycle is dependent on two types of mammalian hosts viz definitive and intermediate hosts. Methods: In this study, enolase, as a key functional enzyme in the metabolism of E. granulosus (EgEnolase), was targeted through a comprehensive in silico modeling analysis and designing a host-specific multi-epitope vaccine. Three-dimensional (3D) structure of enolase was modeled using MODELLER v9.18 software. The B-cell epitopes (BEs) were predicted based on the multi-method approach and via some authentic online predictors. ClusPro v2.0 server was used for docking-based T-helper epitope prediction. The 3D structure of the vaccine was modeled using the RaptorX server. The designed vaccine was evaluated for its immunogenicity, physicochemical properties, and allergenicity. The codon optimization of the vaccine sequence was performed based on the codon usage table of E. coli K12. Finally, the energy minimization and molecular docking were implemented for simulating the vaccine binding affinity to the TLR-2 and TLR-4 and the complex stability. Results: The designed multi-epitope vaccine was found to induce anti-EgEnolase immunity which may have the potential to prevent the survival and proliferation of E. granulosus into the definitive host. Conclusion: Based on the results, this step-by-step immunoinformatics approach could be considered as a rational platform for designing vaccines against such multi-stage parasites. Furthermore, it is proposed that this multi-epitope vaccine is served as a promising preventive anti-echinococcosis agent.

Roadmap to a Successful Product Development

This study details a product development process that accurately predicts the successful implementation of a new product into the market. This product development process evaluates the product development stage of a multi-stage-gate new product development system. It offers a systematic and a rational platform that describes multiple phases that allow for the determination of project viability at the onset of any small to medium product development process. These phases detail interconnected steps with a feedback loop that carry a product concept through a conceptual stage (Phase I) and the generation of a “proof of concept” or an alpha prototype that allows for the initiation of estimates for development cost, market risk analysis, performance evaluation, and product manufacturing cost estimation (Phase II). The final steps are the pre-product design stage (Phase III), which is an expansion of the broader development cost estimates generated in Phase II providing additional rigorous estimates of the product development costs. Notably, modification or redesign of failed product concepts and their conversion into successful prospects that garner market profitability is another tangible objective of the described product development process.

Genome-wide identification and expression analysis of glutathione S-transferase gene family in tomato: Gaining an insight to their physiological and stress-specific roles.

Glutathione S-transferase (GST) refers to one of the major detoxifying enzymes that plays an important role in different abiotic and biotic stress modulation pathways of plant. The present study aimed to a comprehensive genome-wide functional characterization of GST genes and proteins in tomato (Solanum lycopersicum L.). The whole genome sequence analysis revealed the presence of 90 GST genes in tomato, the largest GST gene family reported till date. Eight segmental duplicated gene pairs might contribute significantly to the expansion of SlGST gene family. Based on phylogenetic analysis of tomato, rice, and Arabidopsis GST proteins, GST family members could be further divided into ten classes. Members of each orthologous class showed high conservancy among themselves. Tau and lambda are the major classes of tomato; while tau and phi are the major classes for rice and Arabidopsis. Chromosomal localization revealed highly uneven distribution of SlGST genes in 13 different chromosomes, where chromosome 9 possessed the highest number of genes. Based on publicly available microarray data, expression analysis of 30 available SlGST genes exhibited a differential pattern in all the analyzed tissues and developmental stages. Moreover, most of the members showed highly induced expression in response to multiple biotic and abiotic stress inducers that could be harmonized with the increase in total GST enzyme activity under several stress conditions. Activity of tomato GST could be enhanced further by using some positive modulators (safeners) that have been predicted through molecular docking of SlGSTU5 and ligands. Moreover, tomato GST proteins are predicted to interact with a lot of other glutathione synthesizing and utilizing enzymes such as glutathione peroxidase, glutathione reductase, glutathione synthetase and γ-glutamyltransferase. This comprehensive genome-wide analysis and expression profiling would provide a rational platform and possibility to explore the versatile role of GST genes in crop engineering.

In Silico Dose Prediction for Long-Acting Rilpivirine and Cabotegravir Administration to Children and Adolescents.

Long-acting injectable antiretrovirals represent a pharmacological alternative to oral formulations and an innovative clinical option to address adherence and reduce drug costs. Clinical studies in children and adolescents are characterised by ethical and logistic barriers complicating the identification of dose optimisation. Physiologically-based pharmacokinetic modelling represents a valuable tool to inform dose finding prior to clinical trials. The objective of this study was to simulate potential dosing strategies for existing long-acting injectable depot formulations of cabotegravir and rilpivirine in children and adolescents (aged 3-18years) using physiologically-based pharmacokinetic modelling. Whole-body physiologically-based pharmacokinetic models were developed to represent the anatomical, physiological and molecular processes and age-related changes in children and adolescents through allometric equations. Models were validated for long-acting injectable intramuscular cabotegravir and rilpivirine in adults. Subsequently, the anatomy and physiology of children and adolescents were validated against available literature. The optimal doses of monthly administration of cabotegravir and rilpivirine were identified in children and adolescents, to achieve trough concentrations over the target concentrations derived in a recent efficacy trial of the same formulations. Pharmacokinetic data generated through the physiologically-based pharmacokinetic simulations were similar to observed clinical data in adults. Optimal doses of long-acting injectable antiretrovirals cabotegravir and rilpivirine were predicted using the release rate observed for existing clinical formulations, for different weight groups of children and adolescents. The intramuscular loading dose and maintenance dose of cabotegravir ranged from 200 to 600mg and from 100 to 250mg, respectively, and for rilpivirine it ranged from 250 to 550mg and from 150 to 500mg, respectively, across various weight groups of children ranging from 15 to 70kg. The reported findings represent a rational platform for the identification of suitable dosing strategies and can inform prospective clinical investigation of long-acting injectable formulations in children and adolescents.

Mcl-1 Degradation Is Required for Targeted Therapeutics to Eradicate Colon Cancer Cells.

The Bcl-2 family protein Mcl-1 is often degraded in cancer cells subjected to effective therapeutic treatment, and defective Mcl-1 degradation has been associated with intrinsic and acquired drug resistance. However, a causal relationship between Mcl-1 degradation and anticancer drug responses has not been directly established, especially in solid tumor cells where Mcl-1 inhibition alone is insufficient to trigger cell death. In this study, we present evidence that Mcl-1 participates directly in determining effective therapeutic responses in colon cancer cells. In this setting, Mcl-1 degradation was induced by a variety of multikinase inhibitor drugs, where it relied upon GSK3β phosphorylation and FBW7-dependent ubiquitination. Specific blockade by genetic knock-in (KI) abolished apoptotic responses and conferred resistance to kinase inhibitors. Mcl-1-KI also suppressed the antiangiogenic and anti-hypoxic effects of kinase inhibitors in the tumor microenvironment. Interestingly, these same inhibitors also induced the BH3-only Bcl-2 family protein PUMA, which is required for apoptosis. Degradation-resistant Mcl-1 bound and sequestered PUMA from other prosurvival proteins to maintain cell survival, which was abolished by small-molecule Mcl-1 inhibitors. Our findings establish a pivotal role for Mcl-1 degradation in the response of colon cancer cells to targeted therapeutics, and they provide a useful rational platform to develop Mcl-1-targeting agents that can overcome drug resistance. Cancer Res; 77(9); 2512-21. ©2017 AACR.

Diversification of the urban forest—Can we afford to exclude exotic tree species?

Introduced tree species represent a substantial component of urban forests in cities all over the world. Yet there is controversy about the further use of introduced tree species. Many practice orientated publications,research papers and governmental websites in the fields of urban planning, urban forestry, and urban ecology argue for planting native species and avoiding introduced species. Such arguments for native-only species selection are also touted by environmental groups and the media. Consequently the debate has sometimes spiralled away from a sensible and rational platform where invasion risks and biodiversity loss are discussed, to a groundless and unreasonable argument where exotic species are generally considered incapable of providing ecosystem services. From a European perspective, we here aim to curate a set of necessary considerations for current and future discussions on native and non-native plant material in sustainable urban development. Using examples from Northern and Central Europe we illustrate that in some regions the catalogue of native tree species may be too limited to fulfil ecosystem services and resilience in harsh urban environments. A main message from our line of arguments is that we cannot afford to generally exclude non-native tree species from urban greening. If “native-only” approaches become incorporated in regional, national or international policy documents or legislation there is a risk that urban ecosystem resilience will be compromised, particularly in regions with extreme environmental conditions. Since both invasion risks and sizes of native species pools vary conspicuously at regional to continental scales we also argue to adapt urban policies on using non-native trees to regional contexts.

Generation of Multicellular Tumor Spheroids with Microwell-Based Agarose Scaffolds for Drug Testing

Three dimensional multicellular aggregate, also referred to as cell spheroid or microtissue, is an indispensable tool for in vitro evaluating antitumor activity and drug efficacy. Compared with classical cellular monolayer, multicellular tumor spheroid (MCTS) offers a more rational platform to predict in vivo drug efficacy and toxicity. Nevertheless, traditional processing methods such as plastic dish culture with nonadhesive surfaces are regularly time-consuming, laborious and difficult to provide uniform-sized spheroids, thus causing poor reproducibility of experimental data and impeding high-throughput drug screening. In order to provide a robust and effective platform for in vitro drug evaluation, we present an agarose scaffold prepared with the template containing uniform-sized micro-wells in commercially available cell culture plates. The agarose scaffold allows for good adjustment of MCTS size and large-scale production of MCTS. Transparent agarose scaffold also allows for monitoring of spheroid formation under an optical microscopy. The formation of MCTS from MCF-7 cells was prepared using different-size-well templates and systematically investigated in terms of spheroid growth curve, circularity, and cell viability. The doxorubicin cytotoxicity against MCF-7 spheroid and MCF-7 monolayer cells was compared. The drug penetration behavior, cell cycle distribution, cell apoptosis, and gene expression were also evaluated in MCF-7 spheroid. The findings of this study indicate that, compared with cellular monolayer, MCTS provides a valuable platform for the assessment of therapeutic candidates in an in vivo-mimic microenvironment, and thus has great potential for use in drug discovery and tumor biology research.

Adhesion G protein-coupled receptors are activated by exposure of a cryptic tethered agonist

The large class of adhesion G protein-coupled receptors (aGPCRs) bind extracellular matrix or neighboring cell-surface ligands to regulate organ and tissue development through an unknown activation mechanism. We examined aGPCR activation using two prototypical aGPCRs, GPR56 and GPR110. Active dissociation of the noncovalently bound GPR56 or GPR110 extracellular domains (ECDs) from the respective seven-transmembrane (7TM) domains relieved an inhibitory influence and permitted both receptors to activate defined G protein subtypes. After ECD displacement, the newly revealed short N-terminal stalk regions of the 7TM domains were found to be essential for G protein activation. Synthetic peptides comprising these stalks potently activated GPR56 or GPR110 in vitro or in cells, demonstrating that the stalks comprise a tethered agonist that was encrypted within the ECD. Establishment of an aGPCR activation mechanism provides a rational platform for the development of aGPCR synthetic modulators that could find clinical utility toward aGPCR-directed disease.