Improved glycaemic control induced by evening activity breaks does not persist overnight amongst healthy adults: A randomized crossover trial.

AIMS: To compare the effects of 4 hours of laboratory-based regular activity breaks (RABs) and prolonged sitting (SIT) on subsequent 48-h free-living interstitial glucose levels in a group of healthy adults. MATERIALS AND METHODS: In this randomized crossover trial, participants completed two 4-h laboratory-based interventions commencing at ~5:00 pm: (1) SIT and (2) SIT interrupted with 3 min of body weight resistance exercise activity breaks every 30 min (RABs). Continuous glucose monitoring was performed during the intervention and for 48-h after, during which time participants returned to a free-living setting. RESULTS: Twenty-eight adults (female n = 20, mean ± SD age 25.5 ± 5.6 years, body mass index 29.2 ± 6.9 kg/m2) provided data for this analysis. During the intervention period, RABs lowered mean interstitial glucose by 8.3% (-0.47 mmol/L/4 h, 95% confidence interval [CI] -0.74 to -0.20; p = 0.001) and area under the curve (AUC) by 8.9% (-2.01 mmol/L/4 h, 95% CI -3.05 to -0.97; p < 0.001) compared to SIT. Measures of glycaemic variability were not significantly different during the intervention. There were no significant differences in mean glucose and AUC between conditions during the first nocturnal period and 24-h post intervention. When compared to SIT, RABs increased continuous overall net action of glucose at 1 h and SD glucose by 22% (0.18 mmol/L, 95% CI 0.03 to 0.29; p = 0.018) and 26% (95% CI 4.9 to 42.7; p = 0.019) in the first nocturnal period and by 10% (0.09 mmol/L, 95% CI 0.01, 0.17; p = 0.025) and 15% (95% CI 6.6 to 22.4; p = 0.001) in the 24-h post intervention period, respectively. CONCLUSION: Performing activity breaks in the evening results in acute reductions in interstitial glucose concentrations; however, the magnitude of these changes is not maintained overnight or into the following 48 hours.

Micronutrient status of New Zealand adolescent women consuming vegetarian and non-vegetarian diets.

BACKGROUND AND OBJECTIVES: Globally, there appears to be an ever-increasing interest in adopting a vegetarian diet. However, there are concerns that avoiding meat may increase the risk of anaemia and micronutrient deficiencies, especially for vulnerable populations, such as adolescent women. The objective of this study was to compare the micronutrient status of vegetarian and non-vegetarian adolescent women in New Zealand. METHODS AND STUDY DESIGN: Adolescent women aged 15-18 y were recruited from eight locations across New Zealand. Blood samples were analysed for: haemoglobin, serum ferritin, soluble transferrin receptor, zinc, selenium, retinol binding protein, folate, vitamin B-12, vitamin D and parathyroid hormone. RESULTS: Of the 182 participants who provided a blood sample, 15% self-identified as vegetarian (n=27). On average, vegetarians had 3.1% (95% CI -5.8 to -0.4, p=0.025) lower haemoglobin, and 8.3% (95%CI -14.1 to -2.1, p=0.004) lower selenium. In contrast, serum folate was 80.5% (95% CI 45.7 to 123.7, p<0.001) higher. The prevalence of zinc and selenium deficiency was higher among vegetarians (50% and 12%, respectively) than non-vegetarians (21%, and 2%, respectively). CONCLUSIONS: Adolescent vegetarian women may be at increased risk of deficiency of micronutrients commonly found in animal products, including zinc and selenium, and may benefit from following dietary practices that enhance micronutrient intake and absorption.

Flower-visitor communities of an arcto-alpine plant-Global patterns in species richness, phylogenetic diversity and ecological functioning.

Pollination is an ecosystem function of global importance. Yet, who visits the flower of specific plants, how the composition of these visitors varies in space and time and how such variation translates into pollination services are hard to establish. The use of DNA barcodes allows us to address ecological patterns involving thousands of taxa that are difficult to identify. To clarify the regional variation in the visitor community of a widespread flower resource, we compared the composition of the arthropod community visiting species in the genus Dryas (mountain avens, family Rosaceae), throughout Arctic and high-alpine areas. At each of 15 sites, we sampled Dryas visitors with 100 sticky flower mimics and identified specimens to Barcode Index Numbers (BINs) using a partial sequence of the mitochondrial COI gene. As a measure of ecosystem functioning, we quantified variation in the seed set of Dryas. To test for an association between phylogenetic and functional diversity, we characterized the structure of local visitor communities with both taxonomic and phylogenetic descriptors. In total, we detected 1,360 different BINs, dominated by Diptera and Hymenoptera. The richness of visitors at each site appeared to be driven by local temperature and precipitation. Phylogeographic structure seemed reflective of geological history and mirrored trans-Arctic patterns detected in plants. Seed set success varied widely among sites, with little variation attributable to pollinator species richness. This pattern suggests idiosyncratic associations, with function dominated by few and potentially different taxa at each site. Taken together, our findings illustrate the role of post-glacial history in the assembly of flower-visitor communities in the Arctic and offer insights for understanding how diversity translates into ecosystem functioning.

[Serum 25-hydroxyvitamin D state in healthy children ten year minors old of Barranquilla metropolitan area]. / Estado de la 25-hidroxivitamina D sérica en niños sanos menores de 10 años del área metropolitana de Barranquilla.

OBJETIVE: To evaluate the serum 25-hydroxyvitamin D (25-OH-D) levels in healthy children under 10 years of the Barranquilla metropolitan area. MATERIALS AND METHODS: A descriptive cross-sectional study in which serum levels of 25-OH-D were analyzed in 360 healthy children from 2014 to 2015. RESULTS: The median value of 25-OH-D serum level was 32.23±8.56 ng/mL; 46.38% of children had vitamin D levels in the insufficient range (<30 ng/mL), while 3.05% were deficient (<20 ng/mL). Soledad and Puerto Colombia were the municipalities with more population in this condition. CONCLUSIONS: It is necessary to promote vitamin D supplement consumption and healthy lifestyles in order to safely improve levels of this micronutrient in the population.

OBJETIVO: Evaluar los niveles séricos de 25-hidroxivitamina D (25-OH-D) en niños sanos menores de 10 años del área metropolitana de Barranquilla (AMB). MATERIAL Y MÉTODOS: Estudio descriptivo de corte transversal, que evaluó los niveles séricos de 25-OH-D en 360 niños del AMB en los años 2014-2015. RESULTADOS: El valor promedio de 25-OH-D en la población estudiada fue 32.23±8.56 ng/mL; 46.38% de los niños tenía niveles de vitamina D considerados insuficientes (<30 ng/mL) y 3.05% mostro deficiencia (<20 ng/mL). Soledad y Puerto Colombia fueron los municipios con mayor población en esta condición. CONCLUSIONES: Es necesario generar programas de suplementación nutricional y fomentar estilos de vida que permitan, de forma segura, mejorar los niveles de vitamina D en la población.

Kinetic and structural insights into the binding of histone deacetylase 1 and 2 (HDAC1, 2) inhibitors.

The structure-activity and structure-kinetic relationships of a series of novel and selective ortho-aminoanilide inhibitors of histone deacetylases (HDACs) 1 and 2 are described. Different kinetic and thermodynamic selectivity profiles were obtained by varying the moiety occupying an 11Å channel leading to the Zn(2+) catalytic pocket of HDACs 1 and 2, two paralogs with a high degree of structural similarity. The design of these novel inhibitors was informed by two ligand-bound crystal structures of truncated hHDAC2. BRD4884 and BRD7232 possess kinetic selectivity for HDAC1 versus HDAC2. We demonstrate that the binding kinetics of HDAC inhibitors can be tuned for individual isoforms in order to modulate target residence time while retaining functional activity and increased histone H4K12 and H3K9 acetylation in primary mouse neuronal cell culture assays. These chromatin modifiers, with tuned binding kinetic profiles, can be used to define the relation between target engagement requirements and the pharmacodynamic response of HDACs in different disease applications.

Hydroxamic acid-based histone deacetylase (HDAC) inhibitors can mediate neuroprotection independent of HDAC inhibition.

Histone deacetylase (HDAC) inhibition improves function and extends survival in rodent models of a host of neurological conditions, including stroke, and neurodegenerative diseases. Our understanding, however, of the contribution of individual HDAC isoforms to neuronal death is limited. In this study, we used selective chemical probes to assess the individual roles of the Class I HDAC isoforms in protecting Mus musculus primary cortical neurons from oxidative death. We demonstrated that the selective HDAC8 inhibitor PCI-34051 is a potent neuroprotective agent; and by taking advantage of both pharmacological and genetic tools, we established that HDAC8 is not critically involved in PCI-34051's mechanism of action. We used BRD3811, an inactive ortholog of PCI-34051, and showed that, despite its inability to inhibit HDAC8, it exhibits robust neuroprotective properties. Furthermore, molecular deletion of HDAC8 proved insufficient to protect neurons from oxidative death, whereas both PCI-34051 and BRD3811 were able to protect neurons derived from HDAC8 knock-out mice. Finally, we designed and synthesized two new, orthogonal negative control compounds, BRD9715 and BRD8461, which lack the hydroxamic acid motif and showed that they stably penetrate cell membranes but are not neuroprotective. These results indicate that the protective effects of these hydroxamic acid-containing small molecules are likely unrelated to direct epigenetic regulation via HDAC inhibition, but rather due to their ability to bind metals. Our results suggest that hydroxamic acid-based HDAC inhibitors may mediate neuroprotection via HDAC-independent mechanisms and affirm the need for careful structure-activity relationship studies when using pharmacological approaches.

Evening regular activity breaks extend subsequent free-living sleep time in healthy adults: a randomised crossover trial.

Objective: To determine if performing regular 3-min bouts of resistance exercise spread over 4 hours in an evening will impact subsequent sleep quantity and quality, sedentary time and physical activity compared with prolonged uninterrupted sitting. Methods: In this randomised crossover trial, participants each completed two 4-hour interventions commencing at approximately 17:00 hours: (1) prolonged sitting and (2) sitting interrupted with 3 min of bodyweight resistance exercise activity breaks every 30 min. On completion, participants returned to a free-living setting. This paper reports secondary outcomes relating to sleep quality and quantity, physical activity and sedentary time which were assessed using wrist-worn ActiGraph GT3+ accelerometers paired with a sleep and wear time diary. Results: A total of 28 participants (women, n=20), age 25.6±5.6 years, body mass index 29.5±6.7 kg/m2 (mean±SD) provided data for this analysis. Compared with prolonged sitting, regular activity breaks increased mean sleep period time and time spent asleep by 29.3 min (95% CI: 1.3 to 57.2, p=0.040) and 27.7 min (95% CI: 2.3 to 52.4, p=0.033), respectively, on the night of the intervention. There was no significant effect on mean sleep efficiency (mean: 0.2%, 95% CI: -2.0 to 2.4, p=0.857), wake after sleep onset (1.0 min, 95% CI: -9.6 to 11.7, p=0.849) and number of awakenings (0.8, 95% CI: -1.8 to 3.3, p=0.550). Subsequent 24-hour and 48-hour physical activity patterns were not significantly different. Conclusions: Performing bodyweight resistance exercise activity breaks in the evening has the potential to improve sleep period and total sleep time and does not disrupt other aspects of sleep quality or subsequent 24-hour physical activity. Future research should explore the longer-term impact of evening activity breaks on sleep. Trial registration number: Australian New Zealand Clinical Trials Registry (ACTRN12621000250831).

Breaking Up Evening Sitting with Resistance Activity Improves Postprandial Glycemic Response: A Randomized Crossover Study.

INTRODUCTION: Interrupting sedentary time during the day reduces postprandial glycemia (a risk factor for cardiometabolic disease). However, it is not known if benefits exist for postprandial glucose, insulin and triglyceride responses in the evening, and if these benefits differ by body mass index (BMI) category. METHODS: In a randomized crossover study, 30 participants (25.4 ± 5.4 yr old; BMI 18.5-24.9: n = 10, BMI 25-29.9: n = 10, BMI ≥30: n = 10) completed two intervention arms, beginning at ~1700 h: prolonged sitting for 4 h, and sitting with regular activity breaks of 3 min of resistance exercises every 30 min. Plasma glucose, insulin, and triglyceride concentrations were measured in response to two meals fed at baseline and 120 min. Four-hour incremental area under the curve was compared between interventions. Moderation by BMI status was explored. RESULTS: Overall, when compared with prolonged sitting, regular activity breaks lowered plasma glucose and insulin incremental area under the curve by 31.5% (95% confidence interval = -49.3% to -13.8%) and 26.6% (-39.6% to -9.9%), respectively. No significant differences were found for plasma triglyceride area under the curve. Interactions between BMI status and intervention was not statistically significant. CONCLUSIONS: Interventions that interrupt sedentary time in the evening may improve cardiometabolic health by some magnitude in all participants regardless of bodyweight.

Novel Fluorescence-Based High-Throughput FLIPR Assay Utilizing Membrane-Tethered Genetic Calcium Sensors to Identify T-Type Calcium Channel Modulators.

T-type voltage-gated Ca2+ channels have been implicated in many human disorders, and there has been increasing interest in developing highly selective and potent T-type Ca2+ channel modulators for potential clinical use. However, the unique biophysical properties of T-type Ca2+ channels are not conducive for developing high-throughput screening (HTS) assays to identify modulators, particularly potentiators. To illustrate, T-type Ca2+ channels are largely inactivated and unable to open to allow Ca2+ influx at -25 mV, the typical resting membrane potential of the cell lines commonly used in cellular screening assays. To address this issue, we developed cell lines that express Kir2.3 channels to hyperpolarize the membrane potential to -70 mV, thus allowing T-type channels to return to their resting state where they can be subsequently activated by membrane depolarization in the presence of extracellular KCl. Furthermore, to simplify the HTS assay and to reduce reagent cost, we stably expressed a membrane-tethered genetic calcium sensor, GCaMP6s-CAAX, that displays superior signal to the background compared to the untethered GCaMP6s or the synthetic Ca2+ sensor Fluo-4AM. Here, we describe a novel GCaMP6s-CAAX-based calcium assay utilizing a high-throughput fluorometric imaging plate reader (Molecular Devices, Sunnyvale, CA) format that can identify both activators and inhibitors of T-type Ca2+ channels. Lastly, we demonstrate the utility of this novel fluorescence-based assay to evaluate the activities of two distinct G-protein-coupled receptors, thus expanding the use of GCaMP6s-CAAX to a wide range of applications relevant for developing cellular assays in drug discovery.

Virtual screening for small-molecule pathway regulators by image-profile matching.

Identifying the chemical regulators of biological pathways is a time-consuming bottleneck in developing therapeutics and research compounds. Typically, thousands to millions of candidate small molecules are tested in target-based biochemical screens or phenotypic cell-based screens, both expensive experiments customized to each disease. Here, our uncustomized, virtual, profile-based screening approach instead identifies compounds that match to pathways based on the phenotypic information in public cell image data, created using the Cell Painting assay. Our straightforward correlation-based computational strategy retrospectively uncovered the expected, known small-molecule regulators for 32% of positive-control gene queries. In prospective, discovery mode, we efficiently identified new compounds related to three query genes and validated them in subsequent gene-relevant assays, including compounds that phenocopy or pheno-oppose YAP1 overexpression and kill a Yap1-dependent sarcoma cell line. This image-profile-based approach could replace many customized labor- and resource-intensive screens and accelerate the discovery of biologically and therapeutically useful compounds.

"It's almost expected": rural Australian Aboriginal women's reflections on smoking initiation and maintenance: a qualitative study.

BACKGROUND: Despite declining smoking rates among the general Australian population, rates among Indigenous Australians remain high, with 47% of the Indigenous population reporting daily smoking - twice that of other Australians. Among women, smoking rates are highest in younger age groups, with more than half of Aboriginal women smoking during pregnancy. A lack of research focused on understanding the social context of smoking by Aboriginal women in rural Australia limits our ability to reduce these rates. This study aimed to explore the factors contributing to smoking initiation among rural Aboriginal women and girls and the social context within which smoking behaviour occurs. METHODS: We conducted three focus groups with 14 Aboriginal women and service providers and 22 individual interviews with Aboriginal women from four rural communities to explore their perceptions of the factors contributing to smoking initiation among Aboriginal girls. RESULTS: Four inter-related factors were considered important to understanding the social context in which girls start smoking: colonisation and the introduction of tobacco; normalization of smoking within separate Aboriginal social networks; disadvantage and stressful lives; and the importance of maintaining relationships within extended family and community networks. Within this context, young girls use smoking to attain status and as a way of asserting Aboriginal identity and group membership, a way of belonging, not of rebelling. Family and social structures were seen as providing strong support, but limited the capacity of parents to influence children not to smoke. Marginalization was perceived to contribute to limited aspirations and opportunities, leading to pleasure-seeking in the present rather than having goals for the future. CONCLUSIONS: The results support the importance of addressing contextual factors in any strategies aimed at preventing smoking initiation or supporting cessation among Aboriginal girls and women. It is critical to acknowledge Aboriginal identity and culture as a source of empowerment; and to recognise the role of persistent marginalization in contributing to the high prevalence and initiation of smoking.

The Impact of Organised Sport, Physical Education and Active Commuting on Physical Activity in a Sample of New Zealand Adolescent Females.

BACKGROUND: The majority of adolescents do less physical activity than is recommended by the World Health Organization. Active commuting and participation in organised sport and/or physical education individually have been shown to increase physical activity in adolescents. However, how these domains impact physical activity both individually and in combination has yet to be investigated in a sample of New Zealand female adolescents from around the country. METHODS: Adolescent females aged 15-18 y (n = 111) were recruited from 13 schools across eight locations throughout New Zealand to participate in this cross-sectional study. Participants completed questions about active commuting, and participation in organised sport and physical education, before wearing an Actigraph GT3X (Actigraph, Pensacola, FL, USA) +24 h a day for seven consecutive days to determine time spent in total, MVPA and light physical activity. RESULTS: Active commuters accumulated 17 min/d (95% CI 8 to 26 min/d) more MVPA compared to those who did not. Those who participated in sport accumulated 45 min/d (95% CI 20 to 71 min/d) more light physical activity and 14 min/d (95% CI 5 to 23 min/d) more MVPA compared to those who did not. Participation in physical education did not seem to have a large impact on any component of physical activity. Participation in multiple domains of activity, e.g., active commuting and organised sport, was associated with higher accumulation of MVPA but not light activity. Conclusion Active commuting and sport both contribute a meaningful amount of daily MVPA. Sport participation has the potential to increase overall activity and displace sedentary behaviour. A combination of physical activity domains may be an important consideration when targeting ways to increase physical activity in adolescent females.

Translating hunger training research to primary health: a qualitative study of nurse attitudes towards a novel weight management intervention.

INTRODUCTION Practice nurses in general practice are ideally placed to deliver weight management treatments. Teaching people to eat according to their appetite, based on measurements of blood glucose ('hunger training'), is known to lead to weight loss and improved eating behaviour. To effectively translate this research to primary care requires understanding of key stakeholder perspectives. AIM The aim of this study was to explore the perspectives of practice nurses on the suitability of using hunger training as a weight management intervention in general practice. METHODS Ten nurses trialled hunger training for 1 week, followed by a semi-structured interview where they were asked about their experience; perceived patient interest; enablers and barriers; and suggested changes to hunger training. RESULTS All nurses were positive about hunger training and wanted to use it with their patients. They thought it was a useful method for teaching patients about eating according to their appetite, and the impact of food choices on glucose. Motivation was seen to be both an important potential barrier and enabler for patients. Other anticipated patient enablers included the educational value of hunger training and ease of the programme. Other barriers included lack of time and cost of equipment and appointments. For most nurses, 1 week of following hunger training was sufficient training to deliver the intervention. Suggested refinements included adding nutrition advice to the booklet, incorporating other health goals and enabling social support. DISCUSSION These findings suggest that hunger training could be translated to primary care with minor modifications.

Conservation of the structure and function of bacterial tryptophan synthases.

Tryptophan biosynthesis is one of the most characterized processes in bacteria, in which the enzymes from Salmonella typhimurium and Escherichia coli serve as model systems. Tryptophan synthase (TrpAB) catalyzes the final two steps of tryptophan biosynthesis in plants, fungi and bacteria. This pyridoxal 5'-phosphate (PLP)-dependent enzyme consists of two protein chains, α (TrpA) and ß (TrpB), functioning as a linear αßßα heterotetrameric complex containing two TrpAB units. The reaction has a complicated, multistep mechanism resulting in the ß-replacement of the hydroxyl group of l-serine with an indole moiety. Recent studies have shown that functional TrpAB is required for the survival of pathogenic bacteria in macrophages and for evading host defense. Therefore, TrpAB is a promising target for drug discovery, as its orthologs include enzymes from the important human pathogens Streptococcus pneumoniae, Legionella pneumophila and Francisella tularensis, the causative agents of pneumonia, legionnaires' disease and tularemia, respectively. However, specific biochemical and structural properties of the TrpABs from these organisms have not been investigated. To fill the important phylogenetic gaps in the understanding of TrpABs and to uncover unique features of TrpAB orthologs to spearhead future drug-discovery efforts, the TrpABs from L. pneumophila, F. tularensis and S. pneumoniae have been characterized. In addition to kinetic properties and inhibitor-sensitivity data, structural information gathered using X-ray crystallo-graphy is presented. The enzymes show remarkable structural conservation, but at the same time display local differences in both their catalytic and allosteric sites that may be responsible for the observed differences in catalysis and inhibitor binding. This functional dissimilarity may be exploited in the design of species-specific enzyme inhibitors.

Amino acid anthranilamide derivatives as a new class of glycogen phosphorylase inhibitors.

A series of amino acid anthranilamide derivatives identified from a high-throughput screening campaign as novel, potent, and glucose-sensitive inhibitors of human liver glycogen phosphorylase a are described. A solid-phase synthesis using Wang resin was also developed which provided efficient access to a variety of analogues, and resulted in the identification of key structure-activity relationships, and the discovery of a potent exemplar (IC(50)=80 nM). The SAR scope, synthetic strategy, and in vitro results for this series are presented herein.

High Throughput Screen Identifies Interferon γ-Dependent Inhibitors of Toxoplasma gondii Growth.

Toxoplasma gondii is an obligate intracellular parasite capable of causing severe disease due to congenital infection and in patients with compromised immune systems. Control of infection is dependent on a robust Th1 type immune response including production of interferon gamma (IFN-γ), which is essential for control. IFN-γ activates a variety of antimicrobial mechanisms in host cells, which are then able to control intracellular parasites such as T. gondii. Despite the effectiveness of these pathways in controlling acute infection, the immune system is unable to eradicate chronic infections that can persist for life. Similarly, while antibiotic treatment can control acute infection, it is unable to eliminate chronic infection. To identify compounds that would act synergistically with IFN-γ, we performed a high-throughput screen of diverse small molecule libraries to identify inhibitors of T. gondii. We identified a number of compounds that inhibited parasite growth in vitro at low µM concentrations and that demonstrated enhanced potency in the presence of a low level of IFN-γ. A subset of these compounds act by enhancing the recruitment of light chain 3 (LC3) to the parasite-containing vacuole, suggesting they work by an autophagy-related process, while others were independent of this pathway. The pattern of IFN-γ dependence was shared among the majority of analogs from 6 priority scaffolds, and analysis of structure activity relationships for one such class revealed specific stereochemistry associated with this feature. Identification of these IFN-γ-dependent leads may lead to development of improved therapeutics due to their synergistic interactions with immune responses.

Exploiting an Asp-Glu "switch" in glycogen synthase kinase 3 to design paralog-selective inhibitors for use in acute myeloid leukemia.

Glycogen synthase kinase 3 (GSK3), a key regulatory kinase in the wingless-type MMTV integration site family (WNT) pathway, is a therapeutic target of interest in many diseases. Although dual GSK3α/ß inhibitors have entered clinical trials, none has successfully translated to clinical application. Mechanism-based toxicities, driven in part by the inhibition of both GSK3 paralogs and subsequent ß-catenin stabilization, are a concern in the translation of this target class because mutations and overexpression of ß-catenin are associated with many cancers. Knockdown of GSK3α or GSK3ß individually does not increase ß-catenin and offers a conceptual resolution to targeting GSK3: paralog-selective inhibition. However, inadequate chemical tools exist. The design of selective adenosine triphosphate (ATP)-competitive inhibitors poses a drug discovery challenge due to the high homology (95% identity and 100% similarity) in this binding domain. Taking advantage of an Asp133→Glu196 "switch" in their kinase hinge, we present a rational design strategy toward the discovery of paralog-selective GSK3 inhibitors. These GSK3α- and GSK3ß-selective inhibitors provide insights into GSK3 targeting in acute myeloid leukemia (AML), where GSK3α was identified as a therapeutic target using genetic approaches. The GSK3α-selective compound BRD0705 inhibits kinase function and does not stabilize ß-catenin, mitigating potential neoplastic concerns. BRD0705 induces myeloid differentiation and impairs colony formation in AML cells, with no apparent effect on normal hematopoietic cells. Moreover, BRD0705 impairs leukemia initiation and prolongs survival in AML mouse models. These studies demonstrate feasibility of paralog-selective GSK3α inhibition, offering a promising therapeutic approach in AML.

Functionally Biased D2R Antagonists: Targeting the ß-Arrestin Pathway to Improve Antipsychotic Treatment.

Schizophrenia is a severe neuropsychiatric disease that lacks completely effective and safe therapies. As a polygenic disorder, genetic studies have only started to shed light on its complex etiology. To date, the positive symptoms of schizophrenia are well-managed by antipsychotic drugs, which primarily target the dopamine D2 receptor (D2R). However, these antipsychotics are often accompanied by severe side effects, including motoric symptoms. At D2R, antipsychotic drugs antagonize both G-protein dependent (Gαi/o) signaling and G-protein independent (ß-arrestin) signaling. However, the relevant contributions of the distinct D2R signaling pathways to antipsychotic efficacy and on-target side effects (motoric) are still incompletely understood. Recent evidence from mouse genetic and pharmacological studies point to ß-arrestin signaling as the major driver of antipsychotic efficacy and suggest that a ß-arrestin biased D2R antagonist could achieve an additional level of selectivity at D2R, increasing the therapeutic index of next generation antipsychotics. Here, we characterize BRD5814, a highly brain penetrant ß-arrestin biased D2R antagonist. BRD5814 demonstrated good target engagement via PET imaging, achieving efficacy in an amphetamine-induced hyperlocomotion mouse model with strongly reduced motoric side effects in a rotarod performance test. This proof of concept study opens the possibility for the development of a new generation of pathway selective antipsychotics at D2R with reduced side effect profiles for the treatment of schizophrenia.

An Isochemogenic Set of Inhibitors To Define the Therapeutic Potential of Histone Deacetylases in ß-Cell Protection.

Modulation of histone deacetylase (HDAC) activity has been implicated as a potential therapeutic strategy for multiple diseases. However, it has been difficult to dissect the role of individual HDACs due to a lack of selective small-molecule inhibitors. Here, we report the synthesis of a series of highly potent and isoform-selective class I HDAC inhibitors, rationally designed by exploiting minimal structural changes to the clinically experienced HDAC inhibitor CI-994. We used this toolkit of isochemogenic or chemically matched inhibitors to probe the role of class I HDACs in ß-cell pathobiology and demonstrate for the first time that selective inhibition of an individual HDAC isoform retains beneficial biological activity and mitigates mechanism-based toxicities. The highly selective HDAC3 inhibitor BRD3308 suppressed pancreatic ß-cell apoptosis induced by inflammatory cytokines, as expected, or now glucolipotoxic stress, and increased functional insulin release. In addition, BRD3308 had no effect on human megakaryocyte differentiation, while inhibitors of HDAC1 and 2 were toxic. Our findings demonstrate that the selective inhibition of HDAC3 represents a potential path forward as a therapy to protect pancreatic ß-cells from inflammatory cytokines and nutrient overload in diabetes.

Inhibitors of Glycogen Synthase Kinase 3 with Exquisite Kinome-Wide Selectivity and Their Functional Effects.

The mood stabilizer lithium, the first-line treatment for bipolar disorder, is hypothesized to exert its effects through direct inhibition of glycogen synthase kinase 3 (GSK3) and indirectly by increasing GSK3's inhibitory serine phosphorylation. GSK3 comprises two highly similar paralogs, GSK3α and GSK3ß, which are key regulatory kinases in the canonical Wnt pathway. GSK3 stands as a nodal target within this pathway and is an attractive therapeutic target for multiple indications. Despite being an active field of research for the past 20 years, many GSK3 inhibitors demonstrate either poor to moderate selectivity versus the broader human kinome or physicochemical properties unsuitable for use in in vitro systems or in vivo models. A nonconventional analysis of data from a GSK3ß inhibitor high-throughput screening campaign, which excluded known GSK3 inhibitor chemotypes, led to the discovery of a novel pyrazolo-tetrahydroquinolinone scaffold with unparalleled kinome-wide selectivity for the GSK3 kinases. Taking advantage of an uncommon tridentate interaction with the hinge region of GSK3, we developed highly selective and potent GSK3 inhibitors, BRD1652 and BRD0209, which demonstrated in vivo efficacy in a dopaminergic signaling paradigm modeling mood-related disorders. These new chemical probes open the way for exclusive analyses of the function of GSK3 kinases in multiple signaling pathways involved in many prevalent disorders.