Metabolic liver cancer: associations of rare and common germline variants in one-carbon metabolism and DNA methylation genes.

Animal studies implicate one-carbon metabolism and DNA methylation genes in hepatocellular carcinoma (HCC) development in the setting of metabolic perturbations. Using human samples, we investigated the associations between common and rare variants in these closely related biochemical pathways and risk for metabolic HCC development in a multicenter international study. We performed targeted exome sequencing of 64 genes among 556 metabolic HCC cases and 643 cancer-free controls with metabolic conditions. Multivariable logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs), adjusting for multiple comparisons. Gene-burden tests were used for rare variant associations. Analyses were performed in the overall sample and among non-Hispanic whites. The results show that among non-Hispanic whites, presence of rare functional variants in ABCC2 was associated with 7-fold higher risk of metabolic HCC (OR = 6.92, 95% CI: 2.38-20.15, P = 0.0004), and this association remained significant when analyses were restricted to functional rare variants observed in ≥2 participants (cases 3.2% versus controls 0.0%, P = 1.02 × 10-5). In the overall multiethnic sample, presence of rare functional variants in ABCC2 was nominally associated with metabolic HCC (OR = 3.60, 95% CI: 1.52-8.58, P = 0.004), with similar nominal association when analyses were restricted to functional rare variants observed in ≥2 participants (cases 2.9% versus controls 0.2%, P = 0.006). A common variant in PNPLA3 (rs738409[G]) was associated with higher HCC risk in the overall sample (P = 6.36 × 10-6) and in non-Hispanic whites (P = 0.0002). Our findings indicate that rare functional variants in ABCC2 are associated with susceptibility to metabolic HCC in non-Hispanic whites. PNPLA3-rs738409 is also associated with metabolic HCC risk.

Evaluation of air-free glassware using the ketyl test.

Air-free glassware and techniques are used in most organometallic research laboratories, but their relative effectiveness has not been well studied. We report a method for quantitatively comparing air and moisture exclusion by air-free glassware. Samples of benzophenone dianion in various glassware are monitored by video as their purple color is quenched. Numerical values for time until colorless are extracted from the video data and interpreted as a relative measure of air and moisture exclusion. Low pressure/vacuum (LPV) NMR tubes demonstrated by far the best performance, compared to a variety of Schlenk flasks and standard NMR tubes. The findings are of immediate practical use for synthetic chemists, and the evaluation methods can be used by individual laboratories to assess other conditions for air and moisture exclusion.

Over view of major traumatic injury in Australia--Implications for trauma system design.

BACKGROUND: Trauma registries are known to drive improvements and optimise trauma systems worldwide. This is the first reported comparison of the epidemiology and outcomes at major centres across Australia. METHODS: The Australian Trauma Registry was a collaboration of 26 major trauma centres across Australia at the time of this study and currently collects information on patients admitted to these centres who die after injury and/or sustain major trauma (Injury Severity Score (ISS) > 12). Data from 1 July 2016 to 30 June 2017 were analysed. Primary endpoints were risk adjusted length of stay and mortality (adjusted for age, cause of injury, arrival Glasgow coma scale (GCS), shock-index grouped in quartiles and ISS). RESULTS: There were 8423 patients from 24 centres included. The median age (IQR) was 48 (28-68) years. Median (IQR) ISS was 17 (14-25). There was a predominance of males (72%) apart from the extremes of age. Transport-related cases accounted for 45% of major trauma, followed by falls (35.1%). Patients took 1.42 (1.03-2.12) h to reach hospital and spent 7.10 (3.64-15.00) days in hospital. Risk adjusted length of stay and mortality did not differ significantly across sites. Primary endpoints across sites were also similar in paediatric and older adult (>65) age groups. CONCLUSION: Australia has the capability to identify national injury trends to target prevention and reduce the burden of injury. Quality of care following injury can now be benchmarked across Australia and with the planned enhancements to data collection and reporting, this will enable improved management of trauma victims.

Chlorophyll-a, dissolved organic carbon, turbidity and other variables of ecological importance in river basins in southern Ontario and British Columbia, Canada.

Optical sensing of chlorophyll-a (chl-a), turbidity, and fluorescent dissolved organic matter (fDOM) is often used to characterize the quality of water. There are many site-specific factors and environmental conditions that can affect optically sensed readings; notwithstanding the comparative implication of different procedures used to measure these properties in the laboratory. In this study, we measured these water quality properties using standard laboratory methods, and in the field using optical sensors (sonde-based) at water quality monitoring sites located in four watersheds in Canada. The overall objective of this work was to explore the relationships among sonde-based and standard laboratory measurements of the aforementioned water properties, and evaluate associations among these eco-hydrological properties and land use, environmental, and ancillary water quality variables such as dissolved organic carbon (DOC) and total suspended solids (TSS). Differences among sonde versus laboratory relationships for chl-a suggest such relationships are impacted by laboratory methods and/or site specific conditions. Data mining analysis indicated that interactive site-specific factors predominately impacting chl-a values across sites were specific conductivity and turbidity (variables with positive global associations with chl-a). The overall linear regression predicting DOC from fDOM was relatively strong (R2 = 0.77). However, slope differences in the watershed-specific models suggest laboratory DOC versus fDOM relationships could be impacted by unknown localized water quality properties affecting fDOM readings, and/or the different standard laboratory methods used to estimate DOC. Artificial neural network analyses (ANN) indicated that higher relative chl-a concentrations were associated with low to no tree cover around sample sites and higher daily rainfall in the watersheds examined. Response surfaces derived from ANN indicated that chl-a concentrations were higher where combined agricultural and urban land uses were relatively higher.

Digital detox: The effect of smartphone abstinence on mood, anxiety, and craving.

Whether behavioural addictions should be conceptualised using a similar framework to substance-related addictions remains a topic of considerable debate. Previous literature has developed criteria, which allows any new behavioural addiction to be considered analogous to substance-related addictions. These imply that abstinence from a related object (e.g., smartphones for heavy smartphone users) would lead to mood fluctuations alongside increased levels of anxiety and craving. In a sample of smartphone users, we measured three variables (mood, anxiety, and craving) on four occasions, which included a 24-hour period of smartphone abstinence. Only craving was affected following a short period of abstinence. The results suggest that heavy smartphone usage does not fulfil the criteria required to be considered an addiction. This may have implications for other behavioural addictions.

Conductance quantisation in patterned gate In0.75Ga0.25As structures up to 6 × (2e 2/h).

We present electrical measurements from In0.75Ga0.25As 1D channel devices with Rashba-type, spin-orbit coupling present in the 2D contact regions. Suppressed backscattering as a result of the time-reversal asymmetry at the 1D channel entrance results in enhanced ballistic transport characteristics with clear quantised conductance plateaus up to 6 × (2e 2/h). Applying DC voltages between the source and drain ohmic contacts and an in-plane magnetic field confirms a ballistic transport picture. For asymmetric patterned gate biasing, a lateral spin-orbit coupling effect is weak. However, the Rashba-type spin-orbit coupling leads to a g-factor in the 1D channel that is reduced in magnitude from the 2D value of 9 to ~6.5 in the lowest subband when the effective Rashba field and the applied magnetic field are perpendicular.

Rapid through-container detection of fake spirits and methanol quantification with handheld Raman spectroscopy.

The spirits drinks industry is of significant global economic importance and a major employer worldwide, and the ability to ensure product authenticity and maintain consumer confidence in these high-value products is absolutely essential. Spirit drinks counterfeiting is a worldwide problem, with counterfeiting and adulteration of spirit drinks taking many forms, such as substitution, stretching with lower-grade products, or creation of counterfeits with industrial, surrogate, or locally produced alcohols. Methanol for example, which has been used as a substitute alcohol for ethanol, has a high toxicity in humans. The counterfeiting of spirit drinks is consequently one of the few leading reported types of food fraud which can be directly and unequivocally linked to food safety and health concerns. Here, for the first time, we use handheld Raman spectroscopy with excitation in the near IR (1064 nm) for the through-container differentiation of multiple spirit drinks, detection of multiple chemical markers of counterfeit alcohol, and for the quantification of methanol. We established the limits of detection (LOD) of methanol in the analysed samples from four different spirit types (between 0.23-0.39%), which were considerably lower than a quoted maximum tolerable concentration (MTC) of 2% (v/v) methanol for humans in a 40% alcohol by volume (ABV) spirit drink, and even lower than the general EU limit for naturally occurring methanol in fruit spirits of 0.5% v/v (10 g methanol per L ethanol). We believe that Raman spectroscopy has considerable practicable potential for the rapid in situ through-container detection of counterfeit spirits drinks, as well as for the analysis and protection of other beverages and liquid samples.

Cyclin E Overexpression Sensitizes Triple-Negative Breast Cancer to Wee1 Kinase Inhibition.

PURPOSE: Poor prognosis in triple-negative breast cancer (TNBC) is due to an aggressive phenotype and lack of biomarker-driven targeted therapies. Overexpression of cyclin E and phosphorylated-CDK2 are correlated with poor survival in patients with TNBC, and the absence of CDK2 desensitizes cells to inhibition of Wee1 kinase, a key cell-cycle regulator. We hypothesize that cyclin E expression can predict response to therapies, which include the Wee1 kinase inhibitor, AZD1775. EXPERIMENTAL DESIGN: Mono- and combination therapies with AZD1775 were evaluated in TNBC cell lines and multiple patient-derived xenograft (PDX) models with different cyclin E expression profiles. The mechanism(s) of cyclin E-mediated replicative stress were investigated following cyclin E induction or CRISPR/Cas9 knockout by a number of assays in multiple cell lines. RESULTS: Cyclin E overexpression (i) is enriched in TNBCs with high recurrence rates, (ii) sensitizes TNBC cell lines and PDX models to AZD1775, (iii) leads to CDK2-dependent activation of DNA replication stress pathways, and (iv) increases Wee1 kinase activity. Moreover, treatment of cells with either CDK2 inhibitors or carboplatin leads to transient transcriptional induction of cyclin E (in cyclin E-low tumors) and result in DNA replicative stress. Such drug-mediated cyclin E induction in TNBC cells and PDX models sensitizes them to AZD1775 in a sequential treatment combination strategy.Conclusions: Cyclin E is a potential biomarker of response (i) for AZD1775 as monotherapy in cyclin E-high TNBC tumors and (ii) for sequential combination therapy with CDK2 inhibitor or carboplatin followed by AZD1775 in cyclin E-low TNBC tumors.

Unique interstitial miRNA signature drives fibrosis in a murine model of autosomal dominant polycystic kidney disease.

AIM: To delineate changes in miRNA expression localized to the peri-cystic local microenvironment (PLM) in an orthologous mouse model of autosomal dominant polycystic kidney disease (ADPKD) (mcwPkd1(nl/nl) ). METHODS: We profiled miRNA expression in the whole kidney and laser captured microdissection (LCM) samples from PLM in mcwPkd1(nl/nl) kidneys with Qiagen miScript 384 HC miRNA PCR arrays. The three times points used are: (1) post-natal (PN) day 21, before the development of trichrome-positive areas; (2) PN28, the earliest sign of trichrome staining; and (3) PN42 following the development of progressive fibrosis. PN21 served as appropriate controls and as the reference time point for comparison of miRNA expression profiles. RESULTS: LCM samples revealed three temporally upregulated miRNAs [2 to 2.75-fold at PN28 and 2.5 to 4-fold (P ≤ 0.05) at PN42] and four temporally downregulated miRNAs [2 to 2.75 fold at PN28 and 2.75 to 5-fold (P ≤ 0.05) at PN42]. Expression of twenty-six miRNAs showed no change until PN42 [six decreased (2.25 to 3.5-fold) (P ≤ 0.05) and 20 increased (2 to 4-fold) (P ≤ 0.05)]. Many critical miRNA changes seen in the LCM samples from PLM were not seen in the contralateral whole kidney. CONCLUSION: Precise sampling with LCM identifies miRNA changes that occur with the initiation and progression of renal interstitial fibrosis (RIF). Identification of the target proteins regulated by these miRNAs will provide new insight into the process of fibrosis and identify unique therapeutic targets to prevent or slow the development and progression of RIF in ADPKD.

Tesevatinib ameliorates progression of polycystic kidney disease in rodent models of autosomal recessive polycystic kidney disease.

AIM: To investigate the therapeutic potential of tesevatinib (TSV), a unique multi-kinase inhibitor currently in Phase II clinical trials for autosomal dominant polycystic kidney disease (ADPKD), in well-defined rodent models of autosomal recessive polycystic kidney disease (ARPKD). METHODS: We administered TSV in daily doses of 7.5 and 15 mg/kg per day by I.P. to the well characterized bpk model of polycystic kidney disease starting at postnatal day (PN) 4 through PN21 to assess efficacy and toxicity in neonatal mice during postnatal development and still undergoing renal maturation. We administered TSV by oral gavage in the same doses to the orthologous PCK model (from PN30 to PN90) to assess efficacy and toxicity in animals where developmental processes are complete. The following parameters were assessed: Body weight, total kidney weight; kidney weight to body weight ratios; and morphometric determination of a cystic index and a measure of hepatic disease. Renal function was assessed by: Serum BUN; creatinine; and a 12 h urinary concentrating ability. Validation of reported targets including the level of angiogenesis and inhibition of angiogenesis (active VEGFR2/KDR) was assessed by Western analysis. RESULTS: This study demonstrates that: (1) in vivo pharmacological inhibition of multiple kinase cascades with TSV reduced phosphorylation of key mediators of cystogenesis: EGFR, ErbB2, c-Src and KDR; and (2) this reduction of kinase activity resulted in significant reduction of renal and biliary disease in both bpk and PCK models of ARPKD. The amelioration of disease by TSV was not associated with any apparent toxicity. CONCLUSION: The data supports the hypothesis that this multi-kinase inhibitor TSV may provide an effective clinical therapy for human ARPKD.

Emerging Therapies for Childhood Polycystic Kidney Disease.

Cystic kidney diseases comprise a varied collection of hereditary disorders, where renal cysts comprise a major element of their pleiotropic phenotype. In pediatric patients, the term polycystic kidney disease (PKD) commonly refers to two specific hereditary diseases, autosomal recessive polycystic kidney disease (ARPKD) and autosomal dominant polycystic kidney disease (ADPKD). Remarkable progress has been made in understanding the complex molecular and cellular mechanisms of renal cyst formation in ARPKD and ADPKD. One of the most important discoveries is that both the genes and proteins products of ARPKD and ADPKD interact in a complex network of genetic and functional interactions. These interactions and the shared phenotypic abnormalities of ARPKD and ADPKD, the "cystic phenotypes" suggest that many of the therapies developed and tested for ADPKD may be effective in ARPKD as well. Successful therapeutic interventions for childhood PKD will, therefore, be guided by knowledge of these molecular interactions, as well as a number of clinical parameters, such as the stage of the disease and the rate of disease progression.

Evaluation of an Evidence-Based Weight Loss Trial for Urban African American Adolescents and Caregivers.

Rates of obesity are among the highest for African American adolescents in the US. However, African American adolescents benefit the least from evidence-based weight loss interventions, often experiencing poor treatment retention and low motivation. Participant evaluations provide key information for future development of family-based weight loss interventions able to address these barriers. The purpose of this study was to examine the experiences of African American adolescent and caregivers participating in the FIT Families trial for program satisfaction and content palatability. Content analysis was used to analyze semi-structured exit interviews from 136 African American adolescents [median age 14 years, 69% female] and caregiver pairs [primarily mothers] participating in a family-based 6-month behavioral weight loss intervention that was delivered either in the home or in an office setting. Participants reported most program practices [location, parent involvement, interpersonal relationship with weight loss counselors] and intervention content [cognitive behavioral skills training, motivational interviewing, contingency management] were helpful. Many adolescents [49%] and their caregivers [47%] reported that the program was acceptable overall, however noted that areas for refinement did exist. Participants reported that managing the logistics of weekly sessions was hard. Families expressed a desire for more engaging skills-based learning and the inclusion of exercise sessions and additional tailoring to needs and interests. Individualization, active learning, and support around parenting continues to be beneficial when designing interventions.

The Potato Cryobank at The International Potato Center (Cip): A Model for Long Term Conservation of Clonal Plant Genetic Resources Collections of the Future.

BACKGROUND: Cryobanks are a secure, efficient and low cost method for the long-term conservation of plant genetic resources for theoretically centuries or millennia with minimal maintenance. OBJECTIVE: The present manuscript describes CIP's modified protocol for potato cryopreservation, its large-scale application, and the establishment of quality and operational standards, which included a viability reassessment of material entering the cryobank. MATERIALS AND METHODS: In 2013, CIP established stricter quality and operational standards under which 1,028 potato accessions were cryopreserved with an improved PVS2-droplet protocol. In 2014 the viability of 114 accessions cryopreserved in 2013 accessions were reassessed. RESULTS: The average recovery rate (full plant recovery after LN exposure) of 1028 cryopreserved Solanum species ranged from 34 to 59%, and 70% of the processed accessions showed a minimum recovery rate of ≥20% and were considered as successfully cryopreserved. CONCLUSION: CIP has established a new high quality management system for cryobanking. Periodic viability reassessment, strict and clear recovery criteria and the monitoring of the percent of successful accessions meeting the criteria as well as contamination rates are metrics that need to be considered in cryobanks.

A semiconductor photon-sorter.

Obtaining substantial nonlinear effects at the single-photon level is a considerable challenge that holds great potential for quantum optical measurements and information processing. Of the progress that has been made in recent years one of the most promising methods is to scatter coherent light from quantum emitters, imprinting quantum correlations onto the photons. We report effective interactions between photons, controlled by a single semiconductor quantum dot that is weakly coupled to a monolithic cavity. We show that the nonlinearity of a transition modifies the counting statistics of a Poissonian beam, sorting the photons in number. This is used to create strong correlations between detection events and to create polarization-correlated photons from an uncorrelated stream using a single spin. These results pave the way for semiconductor optical switches operated by single quanta of light.

Attitudes towards HIV testing via home-sampling kits ordered online (RUClear pilots 2011-12).

BACKGROUND: The burden of disease relating to undiagnosed HIV infection is significant in the UK. BHIVA (British HIV Association) recommends population screening in high prevalence areas, expanding outside traditional antenatal/GUM settings. METHODS: RUClear 2011-12 piloted expanding HIV testing outside traditional settings using home-sampling kits (dry-blood-spot testing) ordered online. Greater Manchester residents (≥age 16) could request testing via an established, online chlamydia testing service (www.ruclear.co.uk). Participant attitudes towards this new service were assessed. Qualitative methods (thematic analysis) were used to analyse free-text data submitted by participants via hard copy questionnaires issued in all testing kits. RESULTS: 79.9% (2447/3062) participants completed questionnaires, of which 30.9% (756/2447) provided free-text data. Participants overwhelmingly supported the service, valuing particularly accessibility and convenience, allowing individuals to order tests any time of day and self-sample comfortably at home; avoiding the invasive nature of venipuncture and avoiding the need for face-to-face interaction with health services. The pilot was also clinically and cost-effective. CONCLUSION: Testing via home-sampling kits ordered online (dry-blood-spot testing) was felt to be an acceptable and convenient method for accessing a HIV test. Many individuals undertook HIV testing where they would otherwise not have been tested at all. Expansion of similar services may increase the uptake of HIV testing.

Does standardised structured reporting contribute to quality in diagnostic pathology? The importance of evidence-based datasets.

Key quality parameters in diagnostic pathology include timeliness, accuracy, completeness, conformance with current agreed standards, consistency and clarity in communication. In this review, we argue that with worldwide developments in eHealth and big data, generally, there are two further, often overlooked, parameters if our reports are to be fit for purpose. Firstly, population-level studies have clearly demonstrated the value of providing timely structured reporting data in standardised electronic format as part of system-wide quality improvement programmes. Moreover, when combined with multiple health data sources through eHealth and data linkage, structured pathology reports become central to population-level quality monitoring, benchmarking, interventions and benefit analyses in public health management. Secondly, population-level studies, particularly for benchmarking, require a single agreed international and evidence-based standard to ensure interoperability and comparability. This has been taken for granted in tumour classification and staging for many years, yet international standardisation of cancer datasets is only now underway through the International Collaboration on Cancer Reporting (ICCR). In this review, we present evidence supporting the role of structured pathology reporting in quality improvement for both clinical care and population-level health management. Although this review of available evidence largely relates to structured reporting of cancer, it is clear that the same principles can be applied throughout anatomical pathology generally, as they are elsewhere in the health system.

The role of nurse educators' self-perception and beliefs in the use of learner-centered teaching in the classroom.

This study was designed to investigate the role of nurse educators' beliefs and self-perception in their use of Learner-Centered Teaching in the nursing education classroom. A sample of 122 nurse educators completed an online questionnaire that explored self-perception, beliefs and the correlation with their use of behaviors indicative of Learner-Centered Teaching. Findings demonstrated that nurse educators who identify themselves as learner-centered are more likely to use Learner-Centered Teaching in their classrooms, and nurse educators who strongly believe learner-centered teaching is beneficial in understanding and applying nursing concepts are also somewhat more likely to use Learner-Centered Teaching in the classroom. The results of this study produced a picture of the role of self-perception and beliefs about Learner-Centered Teaching, and the influence this may have on actual utilization of Learner-Centered Teaching. It is clear that valuing Learner-Centered Teaching and believing in its efficacy are only part of the answer to convincing nurse educators to widely adopt this teaching framework.

Electronic Health Records: Describing Technological Stressors of Nurse Educators.

The purpose of this study was to describe the technological stressors that nurse educators experienced when using electronic health records while teaching clinical courses. Survey results indicated that educators had mild to moderate technological stress when teaching the use of electronic health records to students in clinical nursing courses.

Contrast-enhanced CT facilitates rapid, non-destructive assessment of cartilage and bone properties of the human metacarpal.

OBJECTIVE: The aim of this work is to establish the human metacarpal as a new whole joint surface early-stage osteoarthritis (OA) model that enables comparisons of articular cartilage and subchondral bone through high resolution contrast-enhanced CT (CECT) imaging, mechanical testing, and biochemical analysis. DESIGN: The fourth metacarpal was obtained from 12 human cadaveric donors and baseline µCT imaging was followed by indentation testing. The samples were then immersed in anionic (Ioxaglate) and cationic (CA4+) iodinated contrast agent solutions followed by CECT. Cartilage GAG content and distribution was measured using the 1,9 dimethylmethylene blue (DMMB) assay and Safranin-O histology staining. Linear regression was performed to compare cartilage and subchondral bone properties. RESULTS: Strong and significant positive correlations were observed between CA4+ CECT attenuation and both GAG content (R(2) = 0.86) and equilibrium modulus (R(2) = 0.84), while correlations using Ioxaglate were insignificant (R(2) ≤ 0.24, P > 0.05). Subchondral bone plate (SBP) thickness negatively and significantly correlated with SBP mineral density (R(2) = 0.49). Cartilage GAG content significantly correlated with several trabecular bone properties, including positive correlations with bone volume fraction (%BV/TV, R(2) = 0.67), trabecular number (Tb.N, R(2) = 0.60), and trabecular thickness (R(2) = 0.42), and negative relationships with structural model index (SMI, R(2) = 0.78) and trabecular spacing (Tb.Sp, R(2) = 0.56). Similarly, equilibrium modulus correlated positively with %BV/TV (R(2) = 0.50), Tb.N (R(2) = 0.59) and negatively with Tb.Sp (R(2) = 0.55) and SMI (R(2) = 0.60). CONCLUSION: This study establishes the human metacarpal as a new early-stage OA model suitable for rapid, high resolution CECT imaging, mechanical testing, and biochemical analysis of the cartilage and subchondral bone, and for examining their inter-relationships.