Miracle-dz/UniFAR-FTUs huggingface dataset:
Denaturing HPLC of Nucleic Acids

2 Introduction With an increasing number of genome sequences completed, denaturing high performance liquid chromatography (DHPLC) has established itself as one of the most powerful tools for DNA variation screening and allele discrimination (http://insertion.stanford.edu/pub.html). Aside from its obvious application in identifying mutations in human diseases,1 it has provided unprecedented insight into human evolution and prehistoric migrations based on the screening of Y chromosomal and autosomal DNA sequences.2,3 It has been applied successfully to the mapping and cloning of genes in yeast,4 arabidopsis thaliana,5 fruit fly6 and mouse,7 and the screening of chemically induced mutations in a reverse genetic strategy towards understanding the function(s) of genes in arabidopsis8 and fruit fly.9 It has also been implemented in the quantitative measurement of gene expression10 and the analysis of single nucleotide extension products.11 The first step towards the development of DHPLC was taken ten years ago with the synthesis of alkylated non-porous poly(styrene-divinylbenzene) particles 2 microns in diameter.12,13 In combination with a hydroorganic eluent containing alkylammonium ions, this stationary phase material enabled, for the first time, the size-dependent liquid chromatographic separation of double-stranded DNA fragments with a degree of resolution only previously attained by capillary electrophoresis.14 In 1995, it was discovered that double-stranded DNA fragments up to 1500 base pairs in length containing a single-base mismatch, (i.e., a base pair other than one of the two typical Watson-Crick base pairs A-T and G-C, respectively), will be retained less on the aforementioned chromatographic support than a perfectly matched DNA homoduplex at column temperatures sufficiently high to result in partial denaturation of the analytes.15,16 This approach requires that the two sequences to be compared are denatured and reannealed prior to HPLC analysis to allow the formation of the original homoduplices as well as that of heteroduplices between the sense and antisense strands of either homoduplex. Formation of heteroduplices is not required for a more recent embodiment of DHPLC, referred to as completely denaturing HPLC.17 It is based on the ability of the stationary phase to resolve isomeric singlestranded nucleic acids up to 100 nucleotides in length from each other even when they differ only in a single base.
Farnesyltransferase Inhibition Exacerbates Eosinophilic Inflammation and Airway Hyperreactivity in Mice with Experimental Asthma: The Complex Roles of Ras GTPase and Farnesylpyrophosphate in Type 2 Allergic Inflammation

Ras, a small GTPase protein, is thought to mediate Th2-dependent eosinophilic inflammation in asthma. Ras requires cell membrane association for its biological activity, and this requires the posttranslational modification of Ras with an isoprenyl group by farnesyltransferase (FTase) or geranylgeranyltransferase (GGTase). We hypothesized that inhibition of FTase using FTase inhibitor (FTI)–277 would attenuate allergic asthma by depleting membrane-associated Ras. We used the OVA mouse model of allergic inflammation and human airway epithelial (HBE1) cells to determine the role of FTase in inflammatory cell recruitment. BALB/c mice were first sensitized then exposed to 1% OVA aerosol or filtered air, and half were injected daily with FTI-277 (20 mg/kg per day). Treatment of mice with FTI-277 had no significant effect on lung membrane–anchored Ras, Ras protein levels, or Ras GTPase activity. In OVA-exposed mice, FTI-277 treatment increased eosinophilic inflammation, goblet cell hyperplasia, and airway hyperreactivity. Human bronchial epithelial (HBE1) cells were pretreated with 5, 10, or 20 μM FTI-277 prior to and during 12 h IL-13 (20 ng/ml) stimulation. In HBE1 cells, FTase inhibition with FTI-277 had no significant effect on IL-13–induced STAT6 phosphorylation, eotaxin-3 peptide secretion, or Ras translocation. However, addition of exogenous FPP unexpectedly augmented IL-13–induced STAT6 phosphorylation and eotaxin-3 secretion from HBE1 cells without affecting Ras translocation. Pharmacological inhibition of FTase exacerbates allergic asthma, suggesting a protective role for FTase or possibly Ras farnesylation. FPP synergistically augments epithelial eotaxin-3 secretion, indicating a novel Ras-independent farnesylation mechanism or direct FPP effect that promotes epithelial eotaxin-3 production in allergic asthma.
Improv Chat: Second Response Generation for Chatbot

Existing research on response generation for chatbot focuses on \textbf{First Response Generation} which aims to teach the chatbot to say the first response (e.g. a sentence) appropriate to the conversation context (e.g. the user's query). In this paper, we introduce a new task \textbf{Second Response Generation}, termed as Improv chat, which aims to teach the chatbot to say the second response after saying the first response with respect the conversation context, so as to lighten the burden on the user to keep the conversation going. Specifically, we propose a general learning based framework and develop a retrieval based system which can generate the second responses with the users' query and the chatbot's first response as input. We present the approach to building the conversation corpus for Improv chat from public forums and social networks, as well as the neural networks based models for response matching and ranking. We include the preliminary experiments and results in this paper. This work could be further advanced with better deep matching models for retrieval base systems or generative models for generation based systems as well as extensive evaluations in real-life applications.
Problems and Practical Experiences

Prioritisation in public health is more complex than in health services, with permanent tension between ‘purists and pragmatists’. Two types of difficulties are encountered: first, that of unravelling and measuring the values of the society; second, the technical difficulties in determining the expected or probable consequences of different actions. The problem is not only technical, but also it requires us to address conflicts of interest between organisations, and thus to consider problems of implementation and organisational reforms. As health is not the sole aim of social welfare policies, the framework in public health prioritisation should be wide. Ultimately, the decision must be taken as to whether prioritisation should be limited to health, or rather, should address more ambitious – and more difficult – targets in the field of social welfare. By performing prioritisation we fuel the culture of prioritisation, which is essential for its social acceptance. Just as there are orphan drugs, there are also orphan policies, the evaluation of which no organisation is willing to pay. The two most commonly used explicit prioritisation methods are Programme Budgeting and Marginal Analysis (PBMA) and Multi-Criteria Decision Analysis (MCDA). The dimensions that are most frequently applied to health problems or programmes form a long and varied list, and the weights or values assigned to each of these dimensions are of crucial importance in the final ranking obtained. Health plans are well-intentioned attempts at prioritisation, but they are generally unsuccessful because of the technical and political problems outlined above. In general, these initiatives tend to prioritise problems more than solutions, and have evolved towards plans based on the interdepartmental paradigm and towards ‘Health in All Policies’. A checklist for pragmatic considerations in priority setting should include four aspects: (i) clear, well-structured objectives with a clearly-defined time horizon; (ii) an organisation equipped for change; (iii) selecting appropriate people to carry out change; CHAPTER 2
Edinburgh Research Explorer Well-being and arthritis incidence

Wellbeing and Arthritis Abstract Objective : Higher wellbeing is associated with lower levels of inflammatory markers in healthy populations; however, it is unclear whether this association translates into a reduced risk of disease. In the current study, we tested whether the association between wellbeing and inflammation results in a lower risk of arthritis. Methods : The sample consisted of 5,622 participants aged ≥ 50 from the English Longitudinal Study of Ageing and included 6 waves of data collection. We used a structural equation modelling approach to test whether inflammatory markers (C-reactive protein or fibrinogen) mediated the association between wellbeing and arthritis risk over a 10 year follow up period. Results : Higher levels of wellbeing were associated with a decrease in arthritis risk (hazard ratio = 0.97 per unit; 95% confidence interval = 0.96, 0.98; p < 0.001). Of the two inflammatory markers, only C-reactive protein was associated with arthritis risk. Mediation analysis revealed that the indirect effect of wellbeing (at wave 1) on arthritis risk via CRP (at wave 2) was significant (hazard ratio = 0.996; 95% confidence interval = 0.995, 0.998; p < 0.001). This effect remained significant following adjustment for demographic and health behaviour variables and depressive symptoms. Conclusion : C-reactive protein accounts for a small proportion of the association between wellbeing and a reduced risk of arthritis.
High- Versus Low-Dose Statin: Effects on Cardiovascular Events and All-Cause Death

Article, see p 1997 The fact that the concentration of low-density lipoprotein (LDL) cholesterol is a risk factor for atherosclerotic cardiovascular disease (ASCVD) is beyond dispute, with genetic studies indicating that this relationship is causal.1 Furthermore, there is compelling evidence that therapies that reduce the concentration of LDL cholesterol also reduce the risk of having an ASCVD event.2 This evidence is robust in both primary and secondary prevention, in men and women, in older and younger people, in people with and without diabetes mellitus, in people with and without hypertension, and in people with higher or lower levels of baseline LDL cholesterol—in fact, in all people (with the possible exception of those with end-stage renal disease) at risk of having an ASCVD event. There is also evidence in people being treated with a statin that additional lowering of LDL cholesterol, whether by using a higher dose of the same statin,3 by using a more effective statin,4 or by adding a nonstatin LDL cholesterol–lowering agent,5,6 results in a further reduction in the risk of having an ASCVD event. Most of these more versus less LDL cholesterol–lowering studies have been conducted in white populations and have substantially impacted lipid management guidelines in such populations. Until recently, however, there has been no evidence in Asian populations of the clinical benefits of using high versus low doses of a statin. This lack of evidence, combined with a concern of possible adverse effects of high doses of statins in Asians, has resulted in a reluctance to use high doses of statins, even in high-risk Asian people. But there is now compelling evidence that the clinical benefit of using a high versus a …
Investigating Interrelationships among Sales Training Evaluation Methods

Today numerous training options are available to sales organizations, and sales training teams use various means to report training effectiveness. This study utilizes Kirkpatrick’s (1959; 1960) training evaluation model and examines the interrelationships among its four levels of sales training evaluation (i.e., reactions, knowledge acquisition, behavior change, and organizational outcomes). Empirical results indicate that sales trainees’ use of training materials at work is positively related to achieving sales training outcomes, including improving (1) organizational commitment, (2) sales effectiveness, and (3) customer relations. Furthermore, trainees who had positive reactions to training were more likely to learn the material, and trainees with higher levels of knowledge retention were more likely to apply the material in the work environment. Implications are discussed that may aid sales firms to better evaluate training solutions provided by vendors and to develop more effective and accountable sales training efforts.
Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts

Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue that is easily accessible. Through this route, mature neurons can be obtained in a matter of a few weeks. Here, we describe a straightforward and rapid one-step protocol to obtain iNs from dermal fibroblasts obtained through biopsy samples from adult human donors. We explain each step of the process, including the maintenance of the dermal fibroblasts, the freezing procedure to build a stock of the cell line, seeding of the cells for reprogramming, as well as the culture conditions during the conversion process. In addition, we describe the preparation of glass coverslips for electrophysiological recordings, long-term coating conditions, and fluorescence activated cell sorting (FACS). We also illustrate examples of the results to be expected. The protocol described here is easy to perform and can be applied to human fibroblasts derived from human skin biopsies from patients with various different diagnoses and ages. This protocol generates a sufficient amount of iNs which can be used for a wide array of biomedical applications, including disease modeling, drug screening, and target validation.
Stefani Dynamic Ambient Intelligent Sociotechnical Systems Group , Hellenic Open University , Patras , Greece

E-commerce has become a prime channel for doing commerce both globally and locally. It has gained the trust of buyers worldwide not only because of the supporting technologies but for the ever-increasing quality of service provided by most vendors as well. Quality is and will be a competitive advantage in ecommerce as competition on an international scale increases. Designing quality e-commerce software is one battle that needs to be won in this war and tools such as formal standards are key weapons. In this paper an eco-system of software quality metrics is presented based on the ISO25010 standard. These metrics are mapped to quality characteristics of the software and to facets of user-system interaction so as to provide designers and developers insights and guidelines on how produce quality e-commerce systems. A representative set of 29 new metrics for the quality characteristics of Functional Suitability and Usability is presented.
The structure of the TsaB/TsaD/TsaE complex reveals an unexpected mechanism for the bacterial t6A tRNA-modification

Abstract The universal N6-threonylcarbamoyladenosine (t6A) modification at position A37 of ANN-decoding tRNAs is essential for translational fidelity. In bacteria the TsaC enzyme first synthesizes an l-threonylcarbamoyladenylate (TC-AMP) intermediate. In cooperation with TsaB and TsaE, TsaD then transfers the l-threonylcarbamoyl-moiety from TC-AMP onto tRNA. We determined the crystal structure of the TsaB–TsaE–TsaD (TsaBDE) complex of Thermotoga maritima in presence of a non-hydrolysable AMPCPP. TsaE is positioned at the entrance of the active site pocket of TsaD, contacting both the TsaB and TsaD subunits and prohibiting simultaneous tRNA binding. AMPCPP occupies the ATP binding site of TsaE and is sandwiched between TsaE and TsaD. Unexpectedly, the binding of TsaE partially denatures the active site of TsaD causing loss of its essential metal binding sites. TsaE interferes in a pre- or post-catalytic step and its binding to TsaBD is regulated by ATP hydrolysis. This novel binding mode and activation mechanism of TsaE offers good opportunities for antimicrobial drug development.
Low voltage fault ride through control in MMC-HVDC

Modular multilevel converters (MMCs) are the latest converter topology for the high voltage high power applications like the high voltage dc (HVDC) transmission. This paper presents the response of an MMC-HVDC converter during grid faults. Recent grid code requirements specify reactive current injection during low voltage fault conditions and negative sequence current injections during unbalanced faults. The impact of different current control strategies considering the positive and negative sequence current injection upon the MMC converter dynamics are analyzed and its demonstrated through timedomain simulations. It concludes that the oscillations in active power can be minimized at the cost of increased oscillation in the reactive power.
Differentiating Incident from Recurrent Stroke Using Administrative Data: The Impact of Varying Lengths of Look-Back Periods on the Risk of Misclassification

Background/Aims: Administrative data are widely used to monitor epidemiological trends in stroke and outcomes; yet there is scant empirical guidance on how to best differentiate incident from recurrent stroke. Methods: We identified all hospital admissions in New South Wales, Australia, with a principal stroke diagnosis from July 1, 2013 to June 30, 2014, linked to 12 years of previous admissions. We calculated the proportion of cases identified with a prior stroke to determine the number of years of look-back required to minimise misclassification of incident and recurrent strokes. Results: Using the maximum available look-back period of 12 years, 1,171 out of 8,364 eligible stroke cases (14.0%) had a stroke history. A 1-year look-back period identified only 25.1% of these patients and 1 in 10 stroke cases were misclassified as incident. With a 10-year clearance period, less than 1 in 100 stroke cases were misclassified as incident. The risk of misclassification was lower in patients younger than 65 years and in those with haemorrhagic stroke. Conclusion: Hospital administrative data sets linked to prior admissions can be used to distinguish recurrent from incident stroke. The risk of misclassifying recurrent stroke cases as incident events is negligible with a look-back period of 10 years.
Model and energy management system for a parallel hybrid electric unmanned aerial vehicle

In this paper is presented a complete Simulink model and a control strategy for the energy management of a parallel hybrid electric UAV (Unmanned Air Vehicle) powertrain. The model consists of an internal combustion engine, a gearbox (which includes a planetary gear and a continuously variable transmission), an electric motor, which can work also as a generator, an electric drive (Inveter) and a Li-Po battery pack. The control strategy propose a near real-time iterative algorithm based on Dynamic Programming to solve an optimization problem for the optimal power management and torque-split of the powertrain with final state constraints on state variable. Aim of the study is to investigate new flight capabilities that derives from the use of a hybrid architecture, like silent mode using only electric motor, useful for military and civil purpose, and the capacity of reducing fuel consumption. Simulation studies are based on data of an existing UAV and a real flight mission.
Schwann Cell to Axon Transfer of Ribosomes: Toward a Novel Understanding of the Role of Glia in the Nervous System

Schwann cells play pivotal roles in the development and maintenance of the peripheral nervous system. Here, we show that intact sciatic nerve axons of mice contain a small population of ribosomes, which increases by several orders of magnitude when axons are desomatized (severed from their cell bodies). We furthermore demonstrate, using the Wallerian degeneration slow mouse as a model, that Schwann cells transfer polyribosomes to desomatized axons. These data indicate that Schwann cells have the propensity to control axonal protein synthesis by supplying ribosomes on local basis.
Comparison of Different Cuff Pressure Use with the Supreme Laryngeal Mask Airway on Haemodynamic Response, Seal Pressure and Postoperative Adverse Events: A Prospective Randomized Study.

Objective The Supreme™ laryngeal mask airway (SLMA) is a supra glottic airway (SGA) device that is used as an alternative to endotracheal tubes. In the present study, we aimed to compare the use of the SLMA with normal cuff pressure and low cuff pressure, primarily for haemodynamic response. Methods In the present study, 120 patients diagnosed with hypertension and scheduled for varicose vein or inguinal hernia operation were enrolled and 99 patients finished. Using randomization, patients were divided into two groups according to cuff pressure as a low-pressure group (Group L, 45 cm H2O) and a normal-pressure group (Group N, 60 cm H2O). Demographics, Mallampati score and the type and duration of surgery, heart rate (HR), mean arterial pressure (MAP), percentage of tidal volume leakage, Ppeak, Pmean, etCO2, seal pressure, fibreoptic scores and postoperative adverse effects of all patients were recorded. Results MAP and HR values immediately and 2 minutes after SLMA insertion were significantly lower in Group L (p<0.001). In Group L and Group N, the seal pressures were 24.1±3.1 cm H2O and 26.2±3.9 cm H2O, respectively (p=0.003). Also, blood staining and sore throat occurred less frequently in Group L (p<0.05). The fibreoptic average score, insertion features and ventilation parameters were similar between the groups (p>0.05). Conclusion SLMA use with a cuff pressure of 45 cm H2O significantly decreases haemodynamic response and post-operative side effects compared with a normal cuff pressure. Therefore, except for some specific surgeries that require higher seal pressures, we recommend the use of the SLMA with cuff pressures as low as 45 cm H2O.
Allergen Valency, Dose, and FcεRI Occupancy Set Thresholds for Secretory Responses to Pen a 1 and Motivate Design of Hypoallergens

Ag-mediated crosslinking of IgE–FcεRI complexes activates mast cells and basophils, initiating the allergic response. Of 34 donors recruited having self-reported shrimp allergy, only 35% had significant levels of shrimp-specific IgE in serum and measurable basophil secretory responses to rPen a 1 (shrimp tropomyosin). We report that degranulation is linked to the number of FcεRI occupied with allergen-specific IgE, as well as the dose and valency of Pen a 1. Using clustered regularly interspaced palindromic repeat–based gene editing, human RBLrαKO cells were created that exclusively express the human FcεRIα subunit. Pen a 1–specific IgE was affinity purified from shrimp-positive plasma. Cells primed with a range of Pen a 1–specific IgE and challenged with Pen a 1 showed a bell-shaped dose response for secretion, with optimal Pen a 1 doses of 0.1–10 ng/ml. Mathematical modeling provided estimates of receptor aggregation kinetics based on FcεRI occupancy with IgE and allergen dose. Maximal degranulation was elicited when ∼2700 IgE–FcεRI complexes were occupied with specific IgE and challenged with Pen a 1 (IgE epitope valency of ≥8), although measurable responses were achieved when only a few hundred FcεRI were occupied. Prolonged periods of pepsin-mediated Pen a 1 proteolysis, which simulates gastric digestion, were required to diminish secretory responses. Recombinant fragments (60–79 aa), which together span the entire length of tropomyosin, were weak secretagogues. These fragments have reduced dimerization capacity, compete with intact Pen a 1 for binding to IgE–FcεRI complexes, and represent a starting point for the design of promising hypoallergens for immunotherapy.