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Gene sets significantly upregulated upon USP7 inhibitor treatment of H526 cells.

The deubiquitinase USP7 regulates the levels of multiple proteins with roles in cancer progression and immune response. Thus, USP7 inhibition may decrease oncogene function, increase tumor suppressor function, and sensitize tumors to DNA-damaging agents. We have discovered a novel chemical series that potently and selectively inhibits USP7 in biochemical and cellular assays. Our inhibitors reduce the viability of multiple TP53 wild-type cell lines, including several hematologic cancer and MYCN-amplified neuroblastoma cell lines, as well as a subset of TP53-mutant cell lines in vitro. Our work suggests that USP7 inhibitors upregulate transcription of genes normally silenced by the epigenetic repressor complex, polycomb repressive complex 2 (PRC2), and potentiate the activity of PIM and PI3K inhibitors as well as DNA-damaging agents. Furthermore, oral administration of USP7 inhibitors inhibits MM.1S (multiple myeloma; TP53 wild type) and H526 (small cell lung cancer; TP53 mutant) tumor growth in vivo. Our work confirms that USP7 is a promising, pharmacologically tractable target for the treatment of cancer.

Supplementary Figures S1-S6 and legends, and Tables S1, S2, S5, S6, S7.

Societal Impact Statement Tomatoes are important to agriculture, human nutrition and cuisines globally. However, many commercial tomato varieties, including the saladette that dominates the North American market, are highly sensitive to environmental changes that impact yields and critical biochemical pathways including carotenoids and isoprenoids that influence nutritional content and flavour. We highlight the potential of tomato agrodiversity, notably its genetic diversity, as an undervalued research tool for understanding environmental regulation of plant biochemistry under different climates. Yet, tomato genetic diversity in Mexico, the major centre of tomato domestication, is not formally described or protected. We propose that transdisciplinary efforts are essential to identify, conserve and research these globally significant genetic resources.

With very little research exploring intestinal effects of red beetroot consumption, the present pilot study investigated gut microbial changes following red beetroot consumption, via a 14-day intervention trial in healthy adults. Compared to baseline, the study demonstrates transient changes in abundance of some taxa e.g., Romboutsia and Christensenella, after different days of intervention (p 0.05). Enrichment of Akkermansia muciniphila and decrease of Bacteroides fragilis (p 0.05) were observed after 3 days of juice consumption, followed by restoration in abundance after 14 days. With native betacyanins and catabolites detected in stool after juice consumption, betacyanins were found to correlate positively with Bifidobacterium and Coprococcus, and inversely with Ruminococcus (p 0.1), potentiating a significant rise in (iso)butyric acid content (172.7 ± 30.9 µmol/g stool). Study findings indicate the potential of red beetroot to influence gut microbial populations and catabolites associated with these changes, emphasizing the potential benefit of red beetroot on intestinal as well as systemic health.

The deubiquitinase USP7 regulates the levels of multiple proteins with roles in cancer progression and immune response. Thus, USP7 inhibition may decrease oncogene function, increase tumor suppressor function, and sensitize tumors to DNA-damaging agents. We have discovered a novel chemical series that potently and selectively inhibits USP7 in biochemical and cellular assays. Our inhibitors reduce the viability of multiple TP53 wild-type cell lines, including several hematologic cancer and MYCN-amplified neuroblastoma cell lines, as well as a subset of TP53-mutant cell lines in vitro. Our work suggests that USP7 inhibitors upregulate transcription of genes normally silenced by the epigenetic repressor complex, polycomb repressive complex 2 (PRC2), and potentiate the activity of PIM and PI3K inhibitors as well as DNA-damaging agents. Furthermore, oral administration of USP7 inhibitors inhibits MM.1S (multiple myeloma; TP53 wild type) and H526 (small cell lung cancer; TP53 mutant) tumor growth in vivo. Our work confirms that USP7 is a promising, pharmacologically tractable target for the treatment of cancer.

The C-C chemokine receptor 4 (CCR4) is broadly expressed on regulatory T cells (Treg) as well as other circulating and tissue-resident T cells. Treg can be recruited to the tumor microenvironment (TME) through the C-C chemokines CCL17 and CCL22. Treg accumulation in the TME has been shown to dampen the antitumor immune response and is thought to be an important driver in tumor immune evasion. Preclinical and clinical data suggest that reducing the Treg population in the TME can potentiate the antitumor immune response of checkpoint inhibitors. We have developed small-molecule antagonists of CCR4, featuring a novel piperidinyl-azetidine motif, that inhibit the recruitment of Treg into the TME and elicit antitumor responses as a single agent or in combination with an immune checkpoint blockade. The discovery of these potent, selective, and orally bioavailable CCR4 antagonists, and their activity in in vitro and in vivo models, is described herein.

Assessing and managing level of violence risk among forensic mental health patients is a primary role of clinical staff. Forensic risk assessments are typically focused on risk factors and deficits...

Agriculture and food industries generate substantial quantities of waste material with a huge potential for bioactive ingredients to be recovered and converted into high-value chemicals. Red beetroot, known for its high content in betalains, natural red pigments, as well as polyphenols, fiber, and nitrate, is experiencing increasing demand, in particular as juice, which is leaving behind large amounts of waste. The present study focused on the recovery of betalains and polyphenols from dried whole beetroot and wet and dried beet pulp waste from the juicing industry. As part of an ultrasound-assisted extraction, ethanol/water-based solvent mixtures were used as they were found to be more effective than single solvents. Enzyme-assisted extraction was initially examined in the case of wet pulp but was not able to retain betalains. Betalains appear to be more stable in dried pulp. Ultrasound-assisted extraction was found to be more suitable to effectively extract both betalains and polyphenols with a high bioactive yield from dried pulp. The total betalain and polyphenol profiles as well as storage stability and antioxidant capacities were evaluated over a period of four weeks after extraction from the dried waste. During the four-week storage, betalains quickly degraded at room temperature in contrast to −20 °C, whereas polyphenols and antioxidative activity were much less influenced by temperature. When compared, dried samples from the beetroot juicing industry demonstrate good betalain and polyphenol extractability; thus, these data indicate that dried beet waste can serve as a good source of betalains for the color industry and other technological sectors.

Safety is a primary focus in forensic mental health facilities as aggression and violence related to symptoms of mental illness is an ongoing challenge in these settings. Recovery-oriented strategi...

Recruitment of suppressive CD4+ FOXP3+ regulatory T cells (Treg) to the tumor microenvironment (TME) has the potential to weaken the antitumor response in patients receiving treatment with immuno-oncology (IO) agents. Human Treg express CCR4 and can be recruited to the TME through the CC chemokine ligands CCL17 and CCL22. In some cancers, Treg accumulation correlates with poor patient prognosis. Preclinical data suggests that preventing the recruitment of Treg and increasing the population of activated effector T cells (Teff) in the TME can potentiate antitumor immune responses. We developed a novel series of potent, orally bioavailable small molecule antagonists of CCR4. From this series, several compounds exhibited high potency in distinct functional assays in addition to good in vitro and in vivo ADME properties. The design, synthesis, and SAR of this series and confirmation of its in vivo activity are reported.

The present study focused on the development of a new purification protocol suitable for betanin and other major betalains, vulgaxanthin I, indicaxanthin and neobetanin, using flash chromatography which is a convenient and fast method to isolate unstable materials. Following preliminary tests, a gradient procedure using 0-60% acetonitrile, with 0.1% (v/v) formic acid as mobile phase, was selected for the purification. Different fractions were collected based on UV detection at 254 and 280 nm and purities were confirmed by reverse-phase HPLC analysis to be 97%, 95%, 79% and 52% for betanin, indicaxanthin, vulgaxanthin I, and neobetanin, respectively, with pigment yields ranging from 120 to 487 mg per 100 g of powdered raw material. Comparative assessment of antioxidant and radial scavenging properties of individual betalains indicated highest potential for betanin followed by neobetanin, vulgaxanthin I and indicaxanthin.

Background Checkpoint inhibitors (CPIs) such as anti-PD(L)-1 and anti-CTLA-4 antibodies have resulted in unprecedented rates of antitumor responses and extension of survival of patients with a variety of cancers. But some patients fail to respond or initially respond but later relapse as they develop resistance to immune therapy. One of the tumor-extrinsic mechanisms for resistance to immune therapy is the accumulation of regulatory T cells (Treg) in tumors. In preclinical and clinical studies, it has been suggested that tumor trafficking of Treg is mediated by CC chemokine receptor 4 (CCR4). Over 90% of human Treg express CCR4 and migrate toward CCL17 and CCL22, two major CCR4 ligands that are either high at baseline or upregulated in tumors on CPI treatment. Hence, CCR4 antagonism has the potential to be an effective antitumor treatment by reducing the accumulation of Treg into the tumor microenvironment (TME). Methods We developed in vitro and in vivo models to assess Treg migration and antitumor efficacy using a potent and selective CCR4 antagonist, CCR4-351. We used two separate tumor models, Pan02 and CT26 mouse tumors, that have high and low CCR4 ligand expression, respectively. Tumor growth inhibition as well as the frequency of tumor-infiltrating Treg and effector T cells was assessed following the treatment with CCR4 antagonist alone or in combination with CPI. Results Using a selective and highly potent, novel small molecule inhibitor of CCR4, we demonstrate that migration of CCR4+ Treg into the tumor drives tumor progression and resistance to CPI treatment. In tumor models with high baseline levels of CCR4 ligands, blockade of CCR4 reduced the number of Treg and enhanced antitumor immune activity. Notably, in tumor models with low baseline level of CCR4 ligands, treatment with immune CPIs resulted in significant increases of CCR4 ligands and Treg numbers. Inhibition of CCR4 reduced Treg frequency and potentiated the antitumor effects of CPIs. Conclusion Taken together, we demonstrate that CCR4-dependent Treg recruitment into the tumor is an important tumor-extrinsic mechanism for immune resistance. Blockade of CCR4 led to reduced frequency of Treg and resulted in increased antitumor activity, supporting the clinical development of CCR4 inhibitors in combination with CPI for the treatment of cancer. Statement of significance CPI upregulates CCL17 and CCL22 expression in tumors and increases Treg migration into the TME. Pharmacological antagonism of the CCR4 receptor effectively inhibits Treg recruitment and results in enhanced antitumor efficacy either as single agent in CCR4 ligandhigh tumors or in combination with CPIs in CCR4 ligandlow tumors.

BackgroundCheckpoint inhibitors (CPIs) such as anti-PD(L)-1 and anti-CTLA-4 antibodies have resulted in unprecedented rates of antitumor responses and extension of survival of patients with a variety of cancers. But some patients fail to respond or initially respond but later relapse as they develop resistance to immune therapy. One of the tumor-extrinsic mechanisms for resistance to immune therapy is the accumulation of regulatory T cells (Treg) in tumors. In preclinical and clinical studies, it has been suggested that tumor trafficking of Treg is mediated by CC chemokine receptor 4 (CCR4). Over 90% of human Treg express CCR4 and migrate toward CCL17 and CCL22, two major CCR4 ligands that are either high at baseline or upregulated in tumors on CPI treatment. Hence, CCR4 antagonism has the potential to be an effective antitumor treatment by reducing the accumulation of Treg into the tumor microenvironment (TME).MethodsWe developed in vitro and in vivo models to assess Treg migration and antitumor efficacy using a potent and selective CCR4 antagonist, CCR4-351. We used two separate tumor models, Pan02 and CT26 mouse tumors, that have high and low CCR4 ligand expression, respectively. Tumor growth inhibition as well as the frequency of tumor-infiltrating Treg and effector T cells was assessed following the treatment with CCR4 antagonist alone or in combination with CPI.ResultsUsing a selective and highly potent, novel small molecule inhibitor of CCR4, we demonstrate that migration of CCR4+ Treg into the tumor drives tumor progression and resistance to CPI treatment. In tumor models with high baseline levels of CCR4 ligands, blockade of CCR4 reduced the number of Treg and enhanced antitumor immune activity. Notably, in tumor models with low baseline level of CCR4 ligands, treatment with immune CPIs resulted in significant increases of CCR4 ligands and Treg numbers. Inhibition of CCR4 reduced Treg frequency and potentiated the antitumor effects of CPIs.ConclusionTaken together, we demonstrate that CCR4-dependent Treg recruitment into the tumor is an important tumor-extrinsic mechanism for immune resistance. Blockade of CCR4 led to reduced frequency of Treg and resulted in increased antitumor activity, supporting the clinical development of CCR4 inhibitors in combination with CPI for the treatment of cancer.Statement of significanceCPI upregulates CCL17 and CCL22 expression in tumors and increases Treg migration into the TME. Pharmacological antagonism of the CCR4 receptor effectively inhibits Treg recruitment and results in enhanced antitumor efficacy either as single agent in CCR4 ligandhigh tumors or in combination with CPIs in CCR4 ligandlow tumors.

The deubiquitinase USP7 regulates the levels of multiple proteins with roles in cancer progression and immune response. Thus, USP7 inhibition may decrease oncogene function, increase tumor suppressor function, and sensitize tumors to DNA-damaging agents. We have discovered a novel chemical series that potently and selectively inhibits USP7 in biochemical and cellular assays. Our inhibitors reduce the viability of multiple

IntroductionThe availability of β-carotene from provitamin A rich foods in order to improve the provision of vitamin A in humans through food-based approaches is an important factor in mitigating micronutrient deficiencies. However, the extent of intestinal absorption (and subsequent availability of carotenoids to target tissues) is dependent on meal preparation, components of the meal and other intrinsic factors during gastro-intestinal (GI) digestion. Gastric emptying (GE), dictated by the caloric value of a meal, has been suggested as a critical component in determining β-carotene absorption. Indeed, dietary intake of high energy meals may just unravel the underlying factors associated with low uptake of dietary carotenoids during digestion. While several studies have reported the uptake of β-carotene as an individual pure compound under different digestive conditions(1), very few studies have assessed the effects of dietary carotenoids embedded in an energy dense meal, as would normally be consumed, on GI transit time and subsequent nutrient release.Materials and MethodsIn this study, we investigated the role of meal composition, particularly its caloric value, in modulating gastric emptying and thus the absorption of β-carotene. A step-by-step in vitro semi-dynamic model that simulates adult gastric digestion(2) was used on a prepared standard meal whose caloric value was estimated based on the Atwater system as described elsewhere with the assumption that the caloric density of the meal will assume a linear GE rate of 2kcal/min.ResultsPreliminary results indicate that, short transit times of 40 minutes, mimicking early GE, have the highest concentration (3.84 ± 0.014; Mean SD) and therefore, highest bioaccessibility (55.7%) compared to the longer transit time of 160 minutes (0.81 ± 0.002; Mean SD) and (11.8%) for β-carotene concentration and bioaccessibility from a standard meal, respectively.DiscussionThe results suggest that gastric behaviour of the food determines the kinetics of bioaccessibility that may not result in low β-carotene release and ultimately low bioaccessibility from the embedded matrix.

Seasonal variations in crops can alter the profile and amount of constituent compounds and consequentially any biological activity. Differences in phytochemical profile, total phenolic content and inhibitory activity on α-glucosidase (maltase) of Hibiscus sabdariffa calyces grown in South Western Nigeria were determined over wet and dry seasons. The phenolic profile, organic acids and sugars were analysed using HPLC, while inhibition of rat intestinal maltase was measured enzymically. There was a significant increase (1.4-fold; p ≤ 0.05) in total anthocyanin content in the dry compared to wet planting seasons, and maltase inhibition from the dry season was slightly more potent (1.15-fold, p ≤ 0.05). Fructose (1.8-fold), glucose (1.8-fold) and malic acid (3.7-fold) were significantly higher (p ≤ 0.05) but citric acid was lower (62-fold, p ≤ 0.008) in the dry season. Environmental conditions provoke metabolic responses in Hibiscus sabdariffa affecting constituent phytochemicals and nutritional value.

Recovery-orientated care is becoming generally accepted as the best practice, and continued development in the ways it is practiced is necessary to ensure improvement of ongoing care. Forensic pati...

The occurrence of a range of regulated and emerging organic environmental contaminants was investigated in 182 samples of edible marine fish sampled mainly from UK marine regions, but extending northerly to the coast of Norway and south to the Algarve. These species (sprats, mackerel, turbot, halibut, herring, grey mullet, sea bass, grey mullet, sardines, etc.) are among those considered to be at the highest risk of contamination with regulated contaminants such as polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs, dioxins), and polychlorinated biphenyls (PCBs), but the occurrence of polybrominated diphenylethers (PBDEs) and polybrominated biphenyls (PBBs) was also investigated. Sub-sets of samples (50–75) were also analysed for emerging contaminants: polychlorinated naphthalenes (PCNs), polybrominated and mixed halogenated dibenzo-p-dioxins, dibenzofurans and biphenyls (PBDD/Fs, PXDD/Fs and PXBs) and perfluoroalkyl substances (PFAS). Contaminant occurrence varied with species and location, but all measured contaminants were detected, with sprats, sea bass, sardines, mackerel, and herring showing higher tissue concentrations. The concentrations of the different contaminants in the various samples were mapped utilising the GPS coordinate data of the capture locations to visualise spatial distribution levels. In terms of catch location, fish sampled from the coasts of southern Britain, north-western France and the Irish Sea appeared to contain proportionately higher levels of some contaminants - e.g. samples from the Irish Sea tended to show higher PCN concentrations, whereas higher levels of PCBs were observed in some fish sampled off the coasts of northern France. Similarly, samples of mullet from the southeast coast of UK showed much higher concentrations of BDE-99 than the other regions. In terms of occurrence trends, PCDD/F and PCB concentrations show a modest decline over the last decade but where limited background data is available for emerging contaminants, there is no evidence of downward trends. Keywords: PCDD/Fs, PXDD/Fs, PCNs, PBDEs, PFAS, Food, Occurrence trends

Individuals admitted to secure care often experience lengthy hospitalizations and are likely to be admitted on more than one occasion. In the context of growing demand and costs associated with sec...

Bioactive compounds of baobab (Adansonia digitata) pulp from Malawi were investigated. The effect of thermal treatment and storage on selected quality attributes of the juice was also evaluated. Organic compounds were analysed by HPLC; total phenol content (TPC) and total antioxidant activity (FRAP, ABTS and DPPH) were measured by spectrophotometry. Malawi baobab pulp contains high levels of procyanidin B2 (533 ± 22.6 mg/100 g FW), vitamin C (AA + DHA) (466 ± 2.5 mg/100 g FW), gallic acid (68.5 ± 12.4 mg/100 g FW) and (−)-epicatechin (43.0 ± 3.0 mg/100 g FW) and showed a maximum TPC of 1.89 × 103 ± 1.61 mg GAE/100 g FW. The maximum antioxidant activity was 2.81 × 103 ± 92.8 mg TEAC/100 g FW for FRAP, 1.52 × 103 ± 17.1 mg TEAC/100 g FW for ABTS and 50.9 ± 0.43% DPPH for DPPH. Thermal pasteurisation (72 °C, 15 s) retained vitamin C which further showed extended half-life under refrigeration temperature (6 °C). Procyanidin B2, (−)-epicatechin, TPC and antioxidant activity fluctuated during storage. Antioxidant activity was significantly correlated (p ≤ 0.05) with bioactive compounds and TPC.

Recruitment of naturally occurring suppressive CD4+, CD25+, and FOXP3+ regulatory T cells (Treg) to the tumor microenvironment (TME) has the potential to weaken the antitumor response in patients receiving treatment with immuno-oncology (IO) agents. Human Treg express CCR4 and can be recruited to the TME through the C-C chemokines CCL17 and CCL22. We have recently developed a series of potent, orally bioavailable small molecule antagonists of CCR4 that can block recruitment of Treg into the TME.

This article belongs to the Special Issue Dietary Bioactives and Human Health; Around a quarter of the global adult population have metabolic syndrome (MetS) and therefore increased risk of cardiovascular mortality and diabetes. Docosahexaenoic acid, oat beta-glucan and grape anthocyanins have been shown to be effective in reducing MetS risk factors when administered as isolated compounds, but their effect when administered as bioactive-enriched foods has not been evaluated. OBJECTIVE: The overall aim of the PATHWAY-27 project was to evaluate the effectiveness of bioactive-enriched food consumption on improving risk factors of MetS. A pilot study was conducted to assess which of five bioactive combinations provided within three different food matrices (bakery, dairy or egg) were the most effective in adult volunteers. The trial also evaluated the feasibility of production, consumer acceptability and gastrointestinal tolerance of the bioactive-enriched food. METHOD: The study included three monocentric, parallel-arm, double-blind, randomised, dietary intervention trials without a placebo. Each recruiting centre tested the five bioactive combinations within a single food matrix. RESULTS: The study was completed by 167 participants (74 male, 93 female). The results indicated that specific bioactive/matrix combinations have effects on serum triglyceride or HDL-cholesterol level without adverse effects. CONCLUSION: The study evidenced that bioactive-enriched food offers a promising food-based strategy for MetS prevention, and highlighted the importance of conducting pilot studies.

Three varieties of Hibiscus sabdariffa were analyzed for their phytochemical content and inhibitory potential on carbohydrate-digesting enzymes as a basis for selecting a variety for wine production. The dark red variety was chosen as it was highest in phenolic content and an aqueous extract partially inhibited α-glucosidase (maltase), with delphinidin 3-O-sambubioside, cyanidin 3-O-sambubioside, and 3-O-caffeoylquinic acid accounting for 65% of this activity. None of the varieties significantly inhibited α-amylase. Regarding Hibiscus sabdariffa wine, the effect of fermentation temperature (20 and 30 °C) on the physicochemical, phytochemical, and aroma composition was monitored over 40 days. The main change in phytochemical composition observed was the hydrolysis of 3-O-caffeolquinic acid and the concomitant increase of caffeic acid irrespective of fermentation temperature. Wine fermented at 20 °C was slightly more active for α-glucosidase inhibition with more fruity aromas (ethyl octanoate), but there were more flowery notes (2-phenylethanol) at 30 °C.

Individuals with serious mental illness (SMI; i.e., psychotic or major mood disorders) are vulnerable to experiencing multiple forms of adverse safety events in community settings, including violence perpetration and victimization. This study investigates the predictive validity and clinical utility of modifiable risk factors for violence in a sample of 87 forensic psychiatric patients found Not Criminally Responsible on Account of Mental Disorder (NCRMD) transitioning to the community. Using a repeated-measures prospective design, we assessed theoretically based dynamic risk factors (e.g., insight, psychiatric symptoms, negative affect, treatment compliance) before hospital discharge, and at 1 and 6 months postdischarge. Adverse outcomes relevant to this population (e.g., violence, victimization, hospital readmission) were measured at each community follow-up, and at 12 months postdischarge. The base rate of violence (23%) was similar to prior studies of discharged psychiatric patients, but results also highlighted elevated rates of victimization (29%) and hospital readmission (28%) characterizing this sample. Many of the dynamic risk indicators exhibited significant change across time and this change was related to clinically relevant outcomes. Specifically, while controlling for baseline level of risk, fluctuations in dynamic risk factors predicted the likelihood of violence and hospital readmission most consistently (hazard ratios [HR] = 1.35-1.84). Results provide direct support for the utility of dynamic factors in the assessment of violence risk and other adverse community outcomes, and emphasize the importance of incorporating time-sensitive methodologies into predictive models examining dynamic risk. (PsycINFO Database Record

Under nutrient-rich conditions, stochasticity of transcription drives protein expression noise. However, by shifting the environment to amino acid-limited conditions, we identified in E. coli a source of noise whose strength is dictated by translational processes. Specifically, we discovered that cell-to-cell variations in fluorescent protein expression depend on codon choice, with codons yielding lower mean expression after amino acid downshift also resulting in greater noise. We propose that ultra-sensitivity in the tRNA charging/discharging cycle shapes the strength of the observed noise by amplifying fluctuations in global intracellular parameters, such as the concentrations of amino acid, synthetase, and tRNA. We hypothesize that this codon-dependent noise may allow bacteria to selectively optimize cell-to-cell variability in poor environments without relying on low molecular numbers.

Violence and aggression are highly complex problems in mental health care facilities; thus, multi-faceted conflict-reduction strategies are required to mitigate and reduce violence. Safewards is an evidence-informed model aimed at preventing events that have the capacity to trigger aggression and violence. Effectiveness studies of the implementation of Safewards have shown mixed results, including that implementation strategies failed to engage staff and fidelity was low. The objective of this study was to examine the effectiveness of implementing the Safewards model with an approach that embedded co-creation principles in the staff training. Overall, results showed high staff engagement. The average rate of attendance at the classroom-based, staff champion training (n = 108) was 79% (SD = 23). Additionally, online training modules were available to all staff and were completed by 238 of 259 forensic program staff (92%). Overall, staff perceived co-creation to be a positive strategy; staff liked being asked to be involved in the planning, felt that their voices were heard, and believed that it contributed to the success of the Safewards implementation. This study showed that the inclusion of co-creation principles in the implementation strategy enhanced staff adherence to the Safewards model as demonstrated by the high fidelity scores, and effectively led to increased buy-in and engagement of staff.

Orthostatic stress triggers a response to maintain cerebral perfusion and prevent syncope. Given the hypotensive effects of inorganic nitrate this response to orthostasis may be altered by acute supplementation with inorganic nitrate and modified by ethnic origin. Caucasian and SE Asian (n = 30 for both), were recruited and subjected to an ‘active stand test’ and brachial artery blood pressure (BP), digit blood flow and ECG were recorded. Following inorganic nitrate supplementation, (10 mg/kg body mass) the tests were repeated. For both Caucasian and SE Asians transition to standing increased diastolic pressure (DP) and heart rate (HR) (P < 0.001 for both) and by calculation increased rate‐pressure product (P < 0.001) and decreased pulse pressure (P < 0.01 for both) indicative of decreased ventricular filling. Nitrate supplementation decreased both DP (P < 0.001) and HR (P < 0.001). Assessment of HR variability suggested sympathetic nerve activity, was higher throughout in Caucasians (P < 0.05) coupled with higher parasympathetic tone (P < 0.01). Nitrate had no effect on cardiac autonomic nerve activity, as estimated using HR variability, for supine or standing subjects. The tachycardia and hypertension associated with orthostatic stress were preserved in both Caucasian and SE Asian subjects, however, we highlight possible differences in autonomic nervous system activity between Caucasians and SE Asians. SE Asians are resistant to the hypotensive effects of inorganic nitrate supplementation suggesting the absence of a crucial mechanism for activation of the nitrate‐nitrite‐nitric oxide system.

Some commonly consumed marine fish species are considered to display a higher risk of bio-accumulating organic environmental contaminants such as PBDEs. As part of a study to investigate the spatial distribution of these contaminants, data on polybrominated diphenlyethers (PBDEs) and polybrominated biphenyls (PBBs) were collected and analysed by introducing a web-based resource which enables efficient spatial, species and concentration level representations. Furthermore, hierarchical cluster analyses permits correlations within the data to be predicted. The data provide current information on levels of PBDE and PBB occurrence, allowing identification of locations that show higher contaminant levels. 135 fish samples of various species were analysed from UK marine waters, but encompassing the waters around Norway in the North and to the Algarve in the South. PBDEs were observed in all samples with the majority of measured congeners being detected. The concentrations ranged from 0.087 μg/kg to 8.907 μg/kg whole weight (ww) for the sum of all measured PBDE congeners. PBBs occurred less frequently showing a corresponding range of

The effect of temperature (6, 15 and 30 °C) during ageing on the colour, phytochemical composition and bioactivity of roselle wine was investigated over 12 months. At the end of ageing, wines stored at 6 °C had the highest colour density and lowest polymeric anthocyanins. The initial concentration of most of the individual phenolic compounds decreased during ageing, with reduction of monomeric anthocyanins contributing to the formation of anthocyanin-derivatives (pyranoanthocyanins), eight of which were identified tentatively and reported here for the first time in roselle wine. The decrease in individual phenolic compounds did not affect inhibition of α-glucosidase (maltase) activity, which remained relatively low but stable throughout ageing. Diethyl succinate was the only volatile clearly influenced by ageing temperature, with the most pronounced effect at 30 °C (∼256 fold increase). In summary, the final concentrations of anthocyanins and diethyl succinate were the major compounds influenced by ageing temperature.

Upper-limb (UL) recovery following stroke is often poor. UL rehabilitation therefore continues to be a major focus for occupational therapy. Published evidence for the effectiveness of SaeboFlex training in acute stroke rehabilitation is scarce. The purpose of this study is to explore the feasibility and patient experience of SaeboFlex training in acute stroke. This feasibility study recruited stroke patients (

The spectral linewidth of an ensemble of fluorescent emitters is dictated by a combination of the single emitter linewidths and sample inhomogeneities. For semiconductor nanocrystals, efforts to tune ensemble linewidths for optical applications have focused primarily on eliminating sample inhomogeneities because conventional single-molecule methods cannot reliably build accurate ensemble-level statistics for single-particle linewidths. Photon-correlation Fourier spectroscopy in solution (S-PCFS) offers a unique approach to investigating single-nanocrystal spectra with large sample statistics, without user selection bias, with high signal-to-noise ratios, and at fast timescales. With S-PCFS, we directly and quantitatively deconstruct the ensemble linewidth into contributions from the average single-particle linewidth and from sample inhomogeneity. We demonstrate that single-particle linewidths vary significantly from batch to batch and can be synthetically controlled. These findings crystallize our understanding of the synthetic challenges facing underdeveloped nanomaterials such as InP and InAs core/shell particles and introduce new avenues for the synthetic optimization of fluorescent nanoparticles.

This prospective study describes the predictive validity of the Historical Clinical Risk Management-20 Scale (HCR-20) when applied to clinical practice among 109 male mentally disordered offenders in a high secure forensic hospital. Data on violent incidents including reconvictions were collected from multiple sources. The results imply that the implemented HCR-20s did not predict future violence regardless of setting (community vs inpatient) nor time (short vs long term) except for serious incidents. This may indicate that the implemented HCR-20s informed risk management through systematic tailoring of care and treatment plans. Evidence supporting this interpretation was found in a reduction in violent incidents and offending when compared to an earlier study with a similar cohort. Alternatively, the completion of a violence risk assessment by clinicians rather than researchers may have affected the quality of completed assessments. Further research is required to better understand the complex mechanisms...

[EN] The structural properties of oil-in-water (O/W) emulsions, as well as their oxidative stability upon storage at 50 °C, were studied. Eight different formulations were prepared, with the aim of studying the effect of three variables: the composition of the oil phase, the presence of the flavonoid rutin and the homogenization procedure on the structure and the oxidative stability. It was found that high pressure homogenization, through droplet size reduction, stabilized the emulsions both against creaming and oil oxidation. The interfacial protein was also partially replaced by rutin, further improving the stability of the emulsions, whereas purification of the oil phase had hardly any effect. Thus, the structural and oxidative stability of emulsions was controlled by the size of the droplets and improved by the addition of rutin. © 2012 Elsevier Ltd. All rights reserved., Lorena Atares would like to thank the financial support given by the Programa de Apoyo a la Investigacion y Desarrollo (PAID-00-11) from the Universitat Politecnica de Valencia.

BACKGROUND: The goal of limb-sparing surgery for a soft tissue sarcoma of the extremity is to remove all malignant cells while preserving limb function. After initial surgery, microscopic residual disease in the tumor bed will cause a local recurrence in approximately 33% of patients with sarcoma. To help identify these patients, the authors developed an in vivo imaging system to investigate the suitability of molecular imaging for intraoperative visualization. METHODS: A primary mouse model of soft tissue sarcoma and a wide field-of-view imaging device were used to investigate a series of exogenously administered, near-infrared (NIR) fluorescent probes activated by cathepsin proteases for real-time intraoperative imaging. RESULTS: The authors demonstrated that exogenously administered cathepsin-activated probes can be used for image-guided surgery to identify microscopic residual NIR fluorescence in the tumor beds of mice. The presence of residual NIR fluorescence was correlated with microscopic residual sarcoma and local recurrence. The removal of residual NIR fluorescence improved local control. CONCLUSIONS: The authors concluded that their technique has the potential to be used for intraoperative image-guided surgery to identify microscopic residual disease in patients with cancer. Cancer 2012. © 2012 American Cancer Society.

This paper describes the successful implementation of a formal violence risk assessment and management strategy within a high secure forensic care facility. The aim of the implementation was to ensure that each patient had a formal violence risk assessment and management plan that was shared and applied to clinical practice by the patient's clinical team. The process as a whole, from risk assessment to risk management including appropriate care and treatment documentation, is outlined. In this way, this paper also describes the difficulties and problems encountered within the organizational reality of implementation projects. Suggestions and recommendations on how to avoid and manage these are made.

Inpatient aggression in psychiatric settings poses a serious management problem. This study reports the findings of a prospective pilot study on the Dynamic Appraisal of Situational Aggression – Inpatient Version (DASA-IV), a structured risk assessment tool for imminent aggression. The study was conducted in the State Hospital, the high secure psychiatric hospital for Scotland and Northern Ireland. The outcome data were aggressive incidents recorded on the Staff Observation Aggression Scale – Revised (SOAS-R) and incidents noted on the hospital's online recording tool. All measures were completed by nursing staff as part of their daily clinical routine to ensure ecological validity. The DASA-IV was found to be of moderate to good predictive power. Limitations and suggestions for further research are outlined, and the potential for implementation of the tool is discussed.

USP7 is a deubiquitinase (DUB) that has attracted much attention recently due to its multiple roles in promoting cancer progression. By removal of ubiquitin from protein substrates, USP7 stabilizes oncogenes such as MDM2 and Myc, destabilizes and inactivates the key tumor suppressors p53 and PTEN, and imparts resistance to DNA-damaging chemotherapy by enhancing DNA repair responses. USP7 plays an important role in suppression of immune responses in the tumor microenvironment by stabilizing the transcription factor FOXP3 and thereby enhancing the suppressive function of regulatory T cells. Thus, inhibition of USP7 is an appealing therapeutic strategy because it has the potential to impact important oncology targets such as transcription factors that have been widely viewed as undruggable. We employed structure-based and other medicinal chemistry techniques to enable the design of potent and selective USP7 inhibitors. Using a high-throughput assay of DUB activity employing rhodamine-labeled ubiquitin, we optimized several series of reversible USP7 inhibitors to sub-100 pM potency and selectivity of >10,000-fold over all other DUBs. Cellular activity was demonstrated using a luciferase reporter gene assay of p53 activation, revealing compounds with EC50 values ranging down to 20 nM. To assess the role of USP7 inhibition in enhancement of immune responses, we determined relief of suppression of effector T cells in vitro. Effector T cells (CD8+) were co-cultured with regulatory T cells (CD4+ FOXP3+) and antigen-presenting cells for 4 days, after which CD8+ cell proliferation was determined by flow cytometry. Treatment with USP7 inhibitors during co-culture resulted in relief of regulatory T cell suppression of CD8+ cell proliferation. In vivo enhancement of immune responses was assessed in rodent models of inflammation and tumor growth. Direct effects on tumor cell growth and viability were explored by profiling cytotoxicity of USP7 inhibitors as single agents and in combination with chemotherapeutic agents in a broad range of cancer cell lines. In preparation for future clinical development, compounds were modified to obtain desirable in vitro and in vivo ADME and toxicity profiles. Following extensive pre-clinical optimization, we have in hand orally bioavailable compounds with high permeability, low clearance, and minimal off-target activity. Citation Format: Betty Abraham, Lavanya Adusumilli, Berenger Biannic, Delia Bradford, Martin Brovarney, David Chian, Gene Cutler, Xinping Han, Dennis Hu, Scott Jacobson, Sherra Johnson, Paul Kassner, Deepika Kaveri, John Ketcham, Andrea Kim, Paul Leger, Lisa Marshall, Sachie Marubayashi, Jack Maung, Jenny McKinnell, Cesar Meleza, Yamini Ohol, Akinori Okano, Leanne Peiser, Deepa Pookot, Payal Rana, Jacob Schwarz, Nick Shah, Grant Shibuya, Michael Sun, Silpa Suthram, Oezcan Talay, Angela Wadsworth, David Wustrow, Kyle Young, Andrew Napper. Discovery and optimization of potent and selective inhibitors of USP7 to enhance anti-tumor immunity and target tumor growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2915.

We have performed experiments to test whether Epstein Barr Virus (EBV)-infected tumors are enhanced for regulatory T cell (Treg) infiltration and whether selective and potent CCR4 antagonists would be a particularly effective therapeutic in this class of indications. Treg cells, which contribute to an immune-suppressive tumor microenvironment (TME), are attracted to tumors via the recognition of CCL17 and CCL22 ligands by the CCR4 receptor. These chemokines have been shown to be expressed in cells infected by the Epstein Barr Virus (EBV) via the viral LMP1 gene. Tumor types which are frequently associated with EBV-infection include gastric adenocarcinoma (~10% positive), classical Hodgkin's Lymphoma (~50%), and nasopharyngeal carcinoma (~100%). Analyzing RNA expression in EBV-associated tumors, we found strong expression of CCL17, CCL22, and FOXP3, a marker of Treg, when compared to EBV-negative tumors. In fact, NPC tumors show extremely high FOXP3 levels. To further test this link, we obtained EBV-associated tumor samples and performed RNA in situ hybridization (ISH) to measure co-expression of these genes. Strong co-localization, was indeed found, further supporting a link between EBV and Treg recruitment. To directly test whether EBV-positive tumors recruit Treg into tumors via CCL22/17 upregulation, we developed models in which mice were inoculated subcutaneously with EBV-positive cancer cell lines. These EBV-positive cancer cells were assessed for chemokine production in vitro and in vivo by ELISA. We assessed tumor-infiltrating lymphocytes (TILs) in established tumors, including Treg, CD4+ and CD8+ T cells as well as T cell activation markers. Treating these tumor-bearing mice with selective and potent CCR4 small-molecule antagonists alone or in combination with checkpoint-targeting antibodies allowed us to demonstrate meaningful antitumor responses in EBV-positive tumors. Together, these data suggest that EBV-positive tumors, such as gastric adenocarcinomas, Hodgkin's lymphomas, and nasopharyngeal carcinomas, are a class of indications of particular interest and potentially increased responsiveness to small-molecule CCR4 antagonists. These results are helping to inform the ongoing FLX475 trials currently in the clinic. Citation Format: Oezcan Talay, Aparna Jorapur, Scott Jacobson, Sachie Marubayashi, Lisa Marshall, Silpa Suthram, Omar Robles, John Ketcham, Maureen K. Reilly, Ashkaan Younai, Berenger Biannic, Dennis Hu, Minna Bui, Jacob Schwarz, Paul Kassner, Gene Cutler. EBV associated tumors have increased regulatory T cell recruitment and are therefore a potential indication for treatment with potent and selective small molecule CCR4 antagonists [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4752.

Although prima facie evidence for non-ferrous metal-working in the form of slag, crucibles, moulds, etc is present at the Roman town of Calleva Atrebatum at Silchester Hampshire, UK it has not yet been stratigraphically associated with in situ hearths; this can make the location of metal-working difficult to determine. This in turn raises the further question of the range of function of hearths in this urban context. The present study uses a combination of archaeological and analytical methods to attempt to determine the use of several hearth and hearth-related contexts within the on-going excavation at Insula IX. Using X-ray fluorescence (XRF) we were able to distinguish between probable industrial and domestic hearths based on elemental concentrations; for example locally elevated metal concentrations, in particular of copper, show evidence for the probable working of copper alloys at Silchester. Metals were found to accumulate in the silty layers associated with hearth platforms indicating that the tiles themselves do not appear to absorb metal spatter.