Novel carbonic anhydrase IX-targeted therapy enhances the anti-tumour effects of cisplatin in small cell lung cancer.

Small cell lung cancer (SCLC) has an extremely poor prognosis and methods of improving chemotherapeutic intervention are much sought after. A promising approach lies in inhibiting the tumour-associated enzyme, carbonic anhydrase IX (CA IX), which supports tumour cell survival. The aim of this study was to assess the potential of CA IX inhibition using 4-(3'-(3″,5″-dimethylphenyl)ureido)phenyl sulfamate (S4), for the treatment of human SCLC alone and in combination with cisplatin chemotherapy. Treating SCLC cell lines (DMS 79 and COR-L24) with 100 µM S4 reduced viability in vitro and enhanced cell death when combined with 7 µM cisplatin, most prominently under hypoxic conditions (0.1% O2 ). When either cell line was grown as a xenograft tumour in nude mice, intraperitoneal injection of 50 mg/kg S4 alone and in combination with 3 mg/kg cisplatin led to significantly reduced tumour growth. Combination therapy was superior to single agents and response was greatly accentuated when administering repeated doses of cisplatin in DMS 79 tumours. The mechanism of therapeutic response was investigated in vitro, where S4 treatment increased apoptosis under hypoxic conditions in both DMS 79 and COR-L24 cells. DMS 79 tumours receiving S4 in vivo also displayed increased apoptosis and necrosis. Combining S4 with cisplatin reduced both the area of hypoxia and CA IX-positive cells within tumours and increased necrosis, suggesting hypoxia-specific targeting. This study presents a novel, targeted approach to improving current SCLC therapy via inhibition of CA IX, which enhances apoptosis and significantly inhibits xenograft tumour growth when administered alone and in combination with cisplatin chemotherapy.

International survey on high- and low-dose synacthen test and assessment of accuracy in preparing low-dose synacthen.

OBJECTIVE: The short synacthen test (SST) is widely used to assess patients for adrenal insufficiency, but the frequency and protocols used across different centres for the low-dose test (LDT) are unknown. This study aimed to survey centres and test the accuracy of ten different synacthen preparation strategies used for the LDT. METHODS: Members of 6 international endocrine societies were surveyed regarding diagnostic tests used for adrenal insufficiency, and in particular the SST. Synacthen was diluted for the LDT and concentrations measured using a synacthen ELISA. RESULTS: Survey responses were received from 766 individuals across 60 countries (52% adult, 45% paediatric endocrinologists). The SST is used by 98% of centres: 92% using high-dose (250 µg), 43% low-dose and 37% both. Ten low-dose dilution methods were assessed and variation in synacthen concentration was demonstrated with intramethod coefficients of variation (CV) ranging from 2.1% to 109%. The method using 5% dextrose as a diluent was the least variable (CV of 2.1%). The variation in dilution methods means that the dose of synacthen administered in a LDT may vary between 0.16 and 0.81 µg. CONCLUSIONS: The high-dose SST is the most popular diagnostic test of adrenal insufficiency, but up to 72% of paediatric endocrinologists use a LDT. There is considerable variation observed both within and between low-dose synacthen dilution methods creating considerable risk of inaccurate dosing and thereby invalid results.

Loss of hypothalamic Furin affects POMC to proACTH cleavage and feeding behavior in high-fat diet-fed mice.

OBJECTIVE: The hypothalamus regulates feeding and glucose homeostasis through the balanced action of different neuropeptides, which are cleaved and activated by the proprotein convertases PC1/3 and PC2. However, the recent association of polymorphisms in the proprotein convertase FURIN with type 2 diabetes, metabolic syndrome, and obesity, prompted us to investigate the role of FURIN in hypothalamic neurons controlling glucose and feeding. METHODS: POMC-Cre+/- mice were bred with Furinfl/fl mice to generate conditional knockout mice with Furin-deletion in neurons expressing proopiomelanocortin (POMCFurKO), and Furinfl/fl mice were used as controls. POMCFurKO and controls were periodically monitored on both normal chow diet and high fat diet (HFD) for body weight and glucose tolerance by established in-vivo procedures. Food intake was measured in HFD-fed FurKO and controls. Hypothalamic Pomc mRNA was measured by RT-qPCR. ELISAs quantified POMC protein and resulting peptides in the hypothalamic extracts of POMCFurKO mice and controls. The in-vitro processing of POMC was studied by biochemical techniques in HEK293T and CHO cell lines lacking FURIN. RESULTS: In control mice, Furin mRNA levels were significantly upregulated on HFD feeding, suggesting an increased demand for FURIN activity in obesogenic conditions. Under these conditions, the POMCFurKO mice were hyperphagic and had increased body weight compared to Furinfl/fl mice. Moreover, protein levels of POMC were elevated and ACTH concentrations markedly reduced. Also, the ratio of α-MSH/POMC was decreased in POMCFurKO mice compared to controls. This indicates that POMC processing was significantly reduced in the hypothalami of POMCFurKO mice, highlighting for the first time the involvement of FURIN in the cleavage of POMC. Importantly, we found that in vitro, the first stage in processing where POMC is cleaved into proACTH was achieved by FURIN but not by PC1/3 or the other proprotein convertases in cell lines lacking a regulated secretory pathway. CONCLUSIONS: These results suggest that FURIN processes POMC into proACTH before sorting into the regulated secretory pathway, challenging the dogma that PC1/3 and PC2 are the only convertases responsible for POMC cleavage. Furthermore, its deletion affects feeding behaviors under obesogenic conditions.

Understanding Microbial Community Dynamics in Up-Flow Bioreactors to Improve Mitigation Strategies for Oil Souring.

Oil souring occurs when H2S is generated in oil reservoirs. This not only leads to operational risks and health hazards but also increases the cost of refining crude oil. Sulfate-reducing microorganisms are considered to be the main source of the H2S that leads to oil souring. Substrate competition between nitrate-reducing and sulfate-reducing microorganisms makes biosouring mitigation via the addition of nitrate salts a viable strategy. This study explores the shift in microbial community across different phases of biosouring and mitigation. Anaerobic sand-filled columns wetted with seawater and/or oil were used to initiate the processes of sulfidogenesis, followed by mitigation with nitrate, rebound sulfidogenesis, and rebound control phases (via nitrate and low salinity treatment). Shifts in microbial community structure and function were observed across different phases of seawater and oil setups. Marine bacterial taxa (Marinobacter, Marinobacterium, Thalassolituus, Alteromonas, and Cycloclasticus) were found to be the initial responders to the application of nitrate during mitigation of sulfidogenesis in both seawater- and oil- wetted columns. Autotrophic groups (Sulfurimonas and Desulfatibacillum) were found to be higher in seawater-wetted columns compared to oil-wetted columns, suggesting the potential for autotrophic volatile fatty acid (VFA) production in oil-field aquifers when seawater is introduced. Results indicate that fermentative (such as Bacteroidetes) and oil-degrading bacteria (such as Desulfobacula toluolica) play an important role in generating electron donors in the system, which may sustain biosouring and nitrate reduction. Persistence of certain microorganisms (Desulfobacula) across different phases was observed, which may be due to a shift in metabolic lifestyle of the microorganisms across phases, or zonation based on nutrient availability in the columns. Overall results suggest mitigation strategies for biosouring can be improved by monitoring VFA concentrations and microbial community dynamics in the oil reservoirs during secondary recovery of oil.

Potential of using visual imagery to revolutionise measurement of emotional health.

Appropriate measurement of emotional health by all those working with children and young people is an increasing focus for professional practice. Most of the tools used for assessment or self-assessment of emotional health were designed in the mid-20th century using language and technology derived from pen and paper written texts. However, are they fit for purpose in an age of pervasive computing with increasingly rich audiovisual media devices being in the hands of young people? This thought piece explores how the increased use of visual imagery, especially forms that can be viewed or created on digital devices, might provide a way forward for more effective measuring of emotional health, including smiley faces, other emojis and other potential forms of visual imagery. The authors bring together perspectives from healthcare, counselling, youth advocacy, academic research, primary care and school-based mental health support to explore these issues.

Fundamental Movement Skills of Children Living in England: The Role of Ethnicity and Native English Language.

The development of fundamental movement skills (FMS) has been associated with children's general health, and, while there is evidence to suggest that age, gender, physical activity, and socioeconomic status relate to FMS, the relationship of ethnicity and language barriers to FMS competence has been underexplored. These factors may be of particular interest for South Asian (SA) children who have lower physical activity and increased risk of metabolic disease. This cross-sectional study examined ethnic and language differences in FMS among 218 ethnically diverse five-year-old children (61 White ethnic background, 91 SA, 29 Black ethnic background, and 37 other), some with English as a native language ( n = 90) and some with English as an additional language ( n = 75), all recruited from within central England. Each child was assessed performing five locomotor skills (run, gallop, hop, leap, and jump) and six object skills (catch, kick, bounce, strike, underarm roll, and overarm throw) on the Test of Gross Motor Development-2 . A 2 (gender) × 4 (ethnicity) factor analysis of covariance assessed differences in the locomotor and object composite scores and total FMS score, while controlling for body mass index. A two-factor analysis of covariance assessed native language differences in their impact on FMS. We found ethnic and gender differences in FMS ( p < .05) in that SA children had poorer total FMS and locomotor scores than children of either White or Black ethnic backgrounds ( p = .004, p = .001, and p = .008, p = .002, respectively). Girls had poorer total ( p = .001) and locomotor FMS skills ( p < .001). Children with English as an additional language had similar FMS scores compared to children whose native language was English ( p > .05). The findings of low FMS competency in SA children and girls, irrespective of body mass index, suggest that developmentally appropriate interventions targeting SA children and girls are needed in early years. We discuss some unclear factors related to these differences.

Fundamental Motor Skills of Children in Deprived Areas of England: A Focus on Age, Gender and Ethnicity.

This study compared the mastery of fundamental motor skills (FMS) of males and females in early-childhood (four to five years, n = 170) and in middle-childhood (nine to ten years, n = 109) who attend schools in deprived and ethnically diverse areas of England. Process FMS (object control and locomotor skills) were observed using the Test of Gross Motor Development-2. Sprint speed over 10 meters and jump distance assessments were conducted using light gates and tape measures. A gender (male vs. female) by year-group (early-childhood vs. middle-childhood) interaction was shown for the process and product-oriented FMS measurements. Middle-childhood males and females demonstrated significantly greater FMS mastery, as compared to early-childhood (p < 0.05). Furthermore, middle-childhood males demonstrated significantly greater mastery of total FMS, object control skills, and product-oriented assessments, in comparison to females (p < 0.05). Children of Black and White ethnic groups achieved significantly greater mastery of locomotor skills, compared to Asian children, though this did not differ by year-group (p < 0.05). The results suggest that FMS development in deprived and ethnically diverse areas in England varies between genders during middle-childhood and ethnicity. Thus, interventions addressing the lack of FMS mastery achievement, shown in middle-childhood girls and children from Asian ethnic backgrounds, may be pivotal. Further exploration of the role of ethnicity would provide greater clarity in approaching interventions to improve FMS.

The Utility of the Supine-to-Stand Test as a Measure of Functional Motor Competence in Children Aged 5⁻9 Years.

This study examined how supine-to-stand (STS) performance is related to process and product assessment of motor competence (MC) in children. Ninety-one children aged 5⁻9 years were assessed for process and product MC (10 m running speed and standing long jump) as well as process and product measures of STS. Tertiles were created for STS process and STS product scores to create 3 groups reflecting low, medium, and high STS competency. ANCOVA analysis, controlling for age, for process STS, indicated that process MC was significantly higher in children, classified as medium STS (p = 0.048) and high STS (p = 0.011) competence, and that 10 m run speed was slower for low STS compared to medium (p = 0.019) and high STS (p = 0.004). For product STS tertiles, process MC was significantly higher for children in the lowest (fastest) STS tertile compared to those in the medium highest (slowest) tertile (p = 0.01).

Guar Gum Stimulates Biogenic Sulfide Production at Elevated Pressures: Implications for Shale Gas Extraction.

Biogenic sulfide production is a common problem in the oil industry, and can lead to costly hydrocarbon processing and corrosion of extraction infrastructure. The same phenomenon has recently been identified in shale gas extraction by hydraulic fracturing, and organic additives in fracturing fluid have been hypothesized to stimulate this process. Constraining the relative effects of the numerous organic additives on microbial metabolism in situ is, however, extremely challenging. Using a bespoke bioreactor system we sought to assess the potential for guar gum, the most commonly used gelling agent in fracturing fluids, to stimulate biogenic sulfide production by sulfate-reducing microorganisms at elevated pressure. Two pressurized bioreactors were fed with either sulfate-amended freshwater medium, or low-sulfate natural surface water, in addition to guar gum (0.05 w/v%) and an inoculum of sulfate-reducing bacteria for a period of 77 days. Sulfide production was observed in both bioreactors, even when the sulfate concentration was low. Analysis of 16S rRNA gene sequences indicate that heterotrophic bacteria closely associated with the genera Brevundimonas and Acinetobacter became enriched early in the bioreactor experiments, followed by an increase in relative abundance of 16S rRNA genes associated with sulfate-reducing bacteria (Desulfosporosinus and Desulfobacteraceae) at later time points. Results demonstrate that guar gum can stimulate acid- and sulfide-producing microorganisms at elevated pressure, and may have implications for the potential role in microbially induced corrosion during hydraulic fracturing operations. Key differences between experimental and in situ conditions are discussed, as well as additional sources of carbon and energy for biogenic sulfide production during shale gas extraction. Our laboratory approach can be tailored to better simulate deep subsurface conditions in order to probe the role of other fracturing fluid additives and downhole parameters on microbial metabolisms observed in these systems. Such baseline studies will prove essential for effective future development of shale gas worldwide.

Protocol for a cohort study of adolescent mental health service users with a nested cluster randomised controlled trial to assess the clinical and cost-effectiveness of managed transition in improving transitions from child to adult mental health services (the MILESTONE study).

INTRODUCTION: Disruption of care during transition from child and adolescent mental health services (CAMHS) to adult mental health services may adversely affect the health and well-being of service users. The MILESTONE (Managing the Link and Strengthening Transition from Child to Adult Mental Healthcare) study evaluates the longitudinal course and outcomes of adolescents approaching the transition boundary (TB) of their CAMHS and determines the effectiveness of the model of managed transition in improving outcomes, compared with usual care. METHODS AND ANALYSIS: This is a cohort study with a nested cluster randomised controlled trial. Recruited CAMHS have been randomised to provide either (1) managed transition using the Transition Readiness and Appropriateness Measure score summary as a decision aid, or (2) usual care for young people reaching the TB. Participants are young people within 1 year of reaching the TB of their CAMHS in eight European countries; one parent/carer and a CAMHS clinician for each recruited young person; and adult mental health clinician or other community-based care provider, if young person transitions. The primary outcome is Health of the Nation Outcome Scale for Children and Adolescents (HoNOSCA) measuring health and social functioning at 15 months postintervention. The secondary outcomes include mental health, quality of life, transition experience and healthcare usage assessed at 9, 15 and 24 months postintervention. With a mean cluster size of 21, a total of 840 participants randomised in a 1:2 intervention to control are required, providing 89% power to detect a difference in HoNOSCA score of 0.30 SD. The addition of 210 recruits for the cohort study ensures sufficient power for studying predictors, resulting in 1050 participants and an approximate 1:3 randomisation. ETHICS AND DISSEMINATION: The study protocol was approved by the UK National Research Ethics Service (15/WM/0052) and equivalent ethics boards in participating countries. Results will be reported at conferences, in peer-reviewed publications and to all relevant stakeholder groups. TRIAL REGISTRATION NUMBER: ISRCTN83240263; NCT03013595 (pre-results).

The brightness-weight illusion.

Bigger objects look heavier than smaller but otherwise identical objects. When hefted as well as seen, however, bigger objects feel lighter (the size-weight illusion), confirming that the association between visual size and weight has a perceptual component. Darker objects also look heavier than brighter but otherwise identical objects. It is uncertain, however, if this association also has a perceptual element, or if it simply reflects the fact that, in English at least, the same verbal label (light) is applied to both surface brightness and weight. To address this, we looked for a brightness equivalent of the size-weight illusion. Paired-comparison judgments of weight were obtained for balls differing only in color. Based on vision alone, darker objects were judged to be heavier. When the balls were hefted as well as seen, this association was reversed (i.e., a brightness-weight illusion), consistent with it having a perceptual component. To gauge the strength of the illusion (in grams), a white and a black ball (both 129 g) were each compared against a set of mid-gray balls varying in weight. When the balls were hefted as well as seen, the white ball felt approximately 8 g heavier than the black ball, a difference corresponding to 6.2% of their actual weight. Possible environmental origins of the association between surface lightness and weight are considered.