Characterization of three vasopressin receptor 2 variants: an apparent polymorphism (V266A) and two loss-of-function mutations (R181C and M311V).

Arginine vasopressin (AVP) is released from the posterior pituitary and controls water homeostasis. AVP binding to vasopressin V2 receptors (V2Rs) located on kidney collecting duct epithelial cells triggers activation of Gs proteins, leading to increased cAMP levels, trafficking of aquaporin-2 water channels, and consequent increased water permeability and antidiuresis. Typically, loss-of-function V2R mutations cause nephrogenic diabetes insipidus (NDI), whereas gain-of-function mutations cause nephrogenic syndrome of inappropriate antidiuresis (NSIAD). Here we provide further characterization of two mutant V2Rs, R181C and M311V, reported to cause complete and partial NDI respectively, together with a V266A variant, in a patient diagnosed with NSIAD. Our data in HEK293FT cells revealed that for cAMP accumulation, AVP was about 500- or 30-fold less potent at the R181C and M311V mutants than at the wild-type receptor respectively (and about 4000- and 60-fold in COS7 cells respectively). However, in contrast to wild type V2R, the R181C mutant failed to increase inositol phosphate production, while with the M311V mutant, AVP exhibited only partial agonism in addition to a 37-fold potency decrease. Similar responses were detected in a BRET assay for β-arrestin recruitment, with the R181C receptor unresponsive to AVP, and partial agonism with a 23-fold decrease in potency observed with M311V in both HEK293FT and COS7 cells. Notably, the V266A V2R appeared functionally identical to the wild-type receptor in all assays tested, including cAMP and inositol phosphate accumulation, β-arrestin interaction, and in a BRET assay of receptor ubiquitination. Each receptor was expressed at comparable levels. Hence, the M311V V2R retains greater activity than the R181C mutant, consistent with the milder phenotype of NDI associated with this mutant. Notably, the R181C mutant appears to be a Gs protein-biased receptor incapable of signaling to inositol phosphate or recruiting β-arrestin. The etiology of NSIAD in the patient with V266A V2R remains unknown

Discovery of a transient radiation belt at Saturn

Radiation belts have been detected in situ at five planets. Only at Earth however has any variability in their intensity been heretofore observed, in indirect response to solar eruptions and high altitude nuclear explosions. The Cassini spacecraft's MIMI/LEMMS instrument has now detected systematic radiation belt variability elsewhere. We report three sudden increases in energetic ion intensity around Saturn, in the vicinity of the moons Dione and Tethys, each lasting for several weeks, in response to interplanetary events caused by solar eruptions. However, the intensifications, which could create temporary satellite atmospheres at the aforementioned moons, were sharply restricted outside the orbit of Tethys. Unlike Earth, Saturn has almost unchanging inner ion radiation belts: due to Saturn's near-symmetrical magnetic field, Tethys and Dione inhibit inward radial transport of energetic ions, shielding the planet's main, inner radiation belt from solar wind influences

A Review on the Scope of Photothermal Therapy–Based Nanomedicines in Preclinical Models of Colorectal Cancer

Oncologic thermal ablation involves the use of hyperthermic temperatures to damage and treat solid cancers. Thermal ablation is being investigated as a method of treatment in colorectal cancers and has the potential to complement conventional anticancer treatments in managing local recurrence and metastatic disease. Photothermal therapy utilizes photosensitive agents to generate local heat and induce thermal ablation. There is growing interest in developing nanotechnology platforms to deliver such photosensitive agents. An advantage of nanomedicines is their multifunctionality, with the capability to deliver combinations of chemotherapeutics and cancer-imaging agents. To date, there have been no clinical studies evaluating photothermal therapy–based nanomedicines in colorectal cancers. This review presents the current scope of preclinical studies, investigating nanomedicines that have been developed for delivering multimodal photothermal therapy to colorectal cancers, with an emphasis on potential clinical applications

41250 Machine Learning to Identify Predictors of Iatrogenic Injury Using Empirical Bayes Estimates: A Cohort Study of Pressure Injury Prevention

ABSTRACT IMPACT: A machine learning approach using electronic health records can combine descriptive, population-level factors of pressure injury outcomes. OBJECTIVES/GOALS: Pressure injuries cause 60,000 deaths and cost $26 billion annually in the US, but prevention is laborious. We used clinical data to develop a machine learning algorithm for predicting pressure injury risk and prescribe the timing of intervention to help clinicians balance competing priorities. METHODS/STUDY POPULATION: We obtained 94,745 electronic health records with 7,000 predictors to calibrate a predictive algorithm of pressure injury risk. Machine learning was used to mine features predicting changes in pressure injury risk; random forests outperformed neural networks, boosting and bagging in feature selection. These features were fit to multilevel ordered logistic regression to create an algorithm that generated empirical Bayes estimates informing a decision-rule for follow-up based on individual risk trajectories over time. We used cross-validation to verify predictive validity, and constrained optimization to select a best-fit algorithm that reduced the time required to trigger patient follow-up. RESULTS/ANTICIPATED RESULTS: The algorithm significantly improved prediction of pressure injury risk (p<0.001) with an area under the ROC curve of 0.60 compared to the Braden Scale, a traditional clinician instrument of pressure injury risk. At a specificity of 0.50, the model achieved a sensitivity of 0.63 within 2.5 patient-days. Machine learning identified categorical increases in risk when patients were prescribed vasopressors (OR=16.4, p<0.001), beta-blockers (OR=4.8, p<0.001), erythropoietin stimulating agents (OR=3.0, p<0.001), or were ordered a urinalysis screen (OR=9.1, p<0.001), lipid panel (OR=5.7, p<0.001) or pre-albumin panel (OR=2.0, p<0.001). DISCUSSION/SIGNIFICANCE OF FINDINGS: This algorithm could help hospitals conserve resources within a critical period of patient vulnerability for pressure injury not reimbursed by Medicare. Savings generated by this approach could justify investment in machine learning to develop electronic warning systems for many iatrogenic injuries.</jats:p

The role of ABCG2 in modulating responses to anti-cancer photodynamic therapy

The ATP-binding cassette (ABC) superfamily G member 2 (ABCG2) transmembrane protein transporter is known for conferring resistance to treatment in cancers. Photodynamic therapy (PDT) is a promising anti-cancer method involving the use of light-activated photosensitisers to precisely induce oxidative stress and cell death in cancers. ABCG2 can efflux photosensitisers from out of cells, reducing the capacity of PDT and limiting the efficacy of treatment. Many studies have attempted to elucidate the relationship between the expression of ABCG2 in cancers, its effect on the cellular retention of photosensitisers and its impact on PDT. This review looks at the studies which investigate the effect of ABCG2 on a range of different photosensitisers in different pre-clinical models of cancer. This work also evaluates the approaches that are being investigated to address the role of ABCG2 in PDT with an outlook on potential clinical validation

Advanced control strategies for bioprocess chromatography: Challenges and opportunities for intensified processes and next generation products

Recent advances in process analytical technologies and modelling techniques present opportunities to improve industrial chromatography control strategies to enhance process robustness, increase productivity and move towards real-time release testing. This paper provides a critical overview of batch and continuous industrial chromatography control systems for therapeutic protein purification. Firstly, the limitations of conventional industrial fractionation control strategies using in-line UV spectroscopy and on-line HPLC are outlined. Following this, an evaluation of monitoring and control techniques showing promise within research, process development and manufacturing is provided. These novel control strategies combine rapid in-line data capture (e.g. NIR, MALS and variable pathlength UV) with enhanced process understanding obtained from mechanistic and empirical modelling techniques. Finally, a summary of the future states of industrial chromatography control systems is proposed, including strategies to control buffer formulation, product fractionation, column switching and column fouling. The implementation of these control systems improves process capabilities to fulfil product quality criteria as processes are scaled, transferred and operated, thus fast tracking the delivery of new medicines to market

Effectiveness of bridge V.A.C. dressings in the treatment of diabetic foot ulcers

Objectives: This is a prospective study of the clinical efficacy of the V.A.C. Granufoam Bridge Dressing for the treatment of diabetic foot ulcers. Materials and methods: Five consecutive patients with diabetic foot ulcers were treated with V.A.C. Granufoam Bridge Dressings and studied over a period of 22&#x2013;48 days. The indications for treatment included diabetic patients with open ray amputation wounds and wounds post-drainage for abscess with exposed deep structures. Clinical outcome was measured in terms of reduction in wound dimensions, presence of wound granulation, microbial clearance, and development of wound complications. Results: Our results showed that with V.A.C. therapy, wound healing occurred in all patients. The number of dressings required ranged from 8 to 10. The baseline average wound size was 23.1 cm2. Wound areas shrunk by 18.4&#x2013;41.7%. All subjects achieved 100% wound bed granulation with an average length of treatment of 33 days. Microbial clearance was achieved in all cases. All wounds healed by secondary intention in one case and four cases required split-thickness skin grafting. Conclusion: The V.A.C. Granufoam Bridge Dressing is effective in the treatment of diabetic foot ulcers. It promotes reduction of wound area, wound bed granulation, and microbial clearance. By allowing placement of the suction pad outside the foot, it allowed patients to wear protective shoes and to walk non-weight bearing with crutches during V.A.C. therapy

Rule-Based Forecasting: Using Judgment in Time-Series Extrapolation

Rule-Based Forecasting (RBF) is an expert system that uses judgment to develop and apply rules for combining extrapolations. The judgment comes from two sources, forecasting expertise and domain knowledge. Forecasting expertise is based on more than a half century of research. Domain knowledge is obtained in a structured way; one example of domain knowledge is managers= expectations about trends, which we call “causal forces.” Time series are described in terms of 28 conditions, which are used to assign weights to extrapolations. Empirical results on multiple sets of time series show that RBF produces more accurate forecasts than those from traditional extrapolation methods or equal-weights combined extrapolations. RBF is most useful when it is based on good domain knowledge, the domain knowledge is important, the series is well behaved (such that patterns can be identified), there is a strong trend in the data, and the forecast horizon is long. Under ideal conditions, the error for RBF’s forecasts were one-third less than those for equal-weights combining. When these conditions are absent, RBF neither improves nor harms forecast accuracy. Some of RBF’s rules can be used with traditional extrapolation procedures. In a series of studies, rules based on causal forces improved the selection of forecasting methods, the structuring of time series, and the assessment of prediction intervals