Miracle-dz/UniFAR-FTUs huggingface dataset:
Isolation and cultivation of Ca2+ tolerant cardiomyocytes from the adult rat: improvements and applications

1. Primary cultures of cardiomyocytes provide a valuable tool for the study of the pharmacological and toxicological properties of drugs and chemicals, but for several technical reasons cardiomyocytes from adult animals are not routinely used in long-term culture. Because of significant advances in cardiovascular research, tissue engineering and cell transplantation, the need to isolate primary cells from adult animal and/or human tissue is likely to increase in the future. 2. The most common protocols for the isolation and cultivation of cardiomyocytes have been reviewed and the various approaches have been compared. The recent advances in cell culture techniques and the use of the cytoprotective agent, e.g. 2,3-butanedione monoxime greatly increases cell yield and cell viability of isolated and cultured cardiomyocytes. New concepts emerge that enabled an assessment of cellular differentiation in cultured cardiomyocytes and certain specific nuclear transcription factors may play a pivotal role in this process.
Genomics-based approaches to screening carboxylesterase-like genes potentially involved in malathion resistance in oriental migratory locust (Locusta migratoria manilensis).

BACKGROUND Previous studies have indicated that increased carboxylesterase (CarE) activity is a major mechanism of malathion resistance in field populations of the oriental migratory locust, Locusta migratoria manilensis (Meyen), in China. The aim of the present study was to screen CarE-like genes from a large locust expressed sequence tag (EST) database and to assess their potential roles in malathion resistance. RESULTS Twenty-five ESTs derived from different CarE-like genes in the locust EST database were identified, and 12 candidate genes with significantly increased expressions, ranging from 2.6- to 11.6-fold in a field-derived resistant (FR) colony of the locust, were found. These candidate genes were constitutively expressed in all nymph and adult stages, and most of them were predominantly expressed in the gastric caeca and the midgut. Among the 12 genes, two representative genes (LmCarE9 and LmCarE25) were chosen for RNAi followed by malathion bioassay. The nymph mortalities increased from 34.3 to 65.2 and 54.2% respectively after LmCarE9 and LmcarE25 were silenced. These results indicated significant roles of these CarE-like genes in conferring malathion resistance in the locust. CONCLUSION Multiple CarE-like genes were involved in malathion resistance in the locust. As validated by RNAi followed by malathion bioassay, LmCarE9 and LmcarE25 played a significant role in conferring malathion resistance.
Sub-GHz Resonant Magnetoelastic Coupling in Epitaxial Fe Thin Films

The resonant interaction of surface acoustic waves and spin waves is a potential new tool to handle information. However, the natural precession frequencies of most ferromagnetic materials lie in the 10 GHz range, i.e. far above present common technology for surface acoustic waves. This feature restricts the range of materials of interest. To expand this range, we present here measurements of the acoustic attenuation and velocity, at room temperature, in epitaxial Fe on GaAs(001). Resonant coupling is observed in the sub-GHz regime by proper selection of the orientation and intensity of the applied magnetic field. With a very simple magnetization dynamics model that takes into account the softening of the magnetic precession modes, we are able to describe the observed salient features.
Things Are Getting Hairy: Enterobacteria Bacteriophage vB_PcaM_CBB

Enterobacteria phage vB_PcaM_CBB is a “jumbo” phage belonging to the family Myoviridae. It possesses highly atypical whisker-like structures along the length of its contractile tail. It has a broad host range with the capability of infecting species of the genera Erwinia, Pectobacterium, and Cronobacter. With a genome of 355,922 bp, excluding a predicted terminal repeat of 22,456 bp, phage CBB is the third largest phage sequenced to date. Its genome was predicted to encode 554 ORFs with 33 tRNAs. Based on prediction and proteome analysis of the virions, 29% of its predicted ORFs could be functionally assigned. Protein comparison shows that CBB shares between 33–38% of its proteins with Cronobacter phage GAP32, coliphages PBECO4 and 121Q as well as Klebsiella phage vB_KleM_Rak2. This work presents a detailed and comparative analysis of vB_PcaM_CBB of a highly atypical jumbo myoviridae phage, contributing to a better understanding of phage diversity and biology.
PTP-PEST targets a novel tyrosine site in p120 catenin to control epithelial cell motility and Rho GTPase activity

ABSTRACT Tyrosine phosphorylation is implicated in regulating the adherens junction protein, p120 catenin (p120), however, the mechanisms are not well defined. Here, we show, using substrate trapping, that p120 is a direct target of the protein tyrosine phosphatase, PTP-PEST, in epithelial cells. Stable shRNA knockdown of PTP-PEST in colon carcinoma cells results in an increased cytosolic pool of p120 concomitant with its enhanced tyrosine phosphorylation and decreased association with E-cadherin. Consistent with this, PTP-PEST knockdown cells exhibit increased motility, enhanced Rac1 and decreased RhoA activity on a collagen substrate. Furthermore, p120 localization is enhanced at actin-rich protrusions and lamellipodia and has an increased association with the guanine nucleotide exchange factor, VAV2, and cortactin. Exchange factor activity of VAV2 is enhanced by PTP-PEST knockdown whereas overexpression of a VAV2 C-terminal domain or DH domain mutant blocks cell motility. Analysis of point mutations identified tyrosine 335 in the N-terminal domain of p120 as the site of PTP-PEST dephosphorylation. A Y335F mutant of p120 failed to induce the ‘p120 phenotype’, interact with VAV2, stimulate cell motility or activate Rac1. Together, these data suggest that PTP-PEST affects epithelial cell motility by controlling the distribution and phosphorylation of p120 and its availability to control Rho GTPase activity.
Phase II trial of single-agent oral vinorelbine in elderly (> or =70 years) patients with advanced non-small-cell lung cancer and poor performance status.

BACKGROUND Elderly patients with advanced non-small-cell lung cancer (NSCLC) with poor performance status (PS) are a special population requiring particular attention. Single-agent oral vinorelbine could be an attractive option. PATIENTS AND METHODS A total of 43 patients with stage IIIB-IV NSCLC and Eastern Cooperative Oncology Group (ECOG) PS of two or more with good functional status were prospectively recruited. Oral vinorelbine was administered at the dose of 60 mg/m(2) on days 1-8 every 3 weeks. Primary end points were response rate and safety. RESULTS Overall response rate was 18.6% with 8 partial responses; 18 of 43 (41.8%) experienced stable disease lasting >12 weeks and 17 of 43 (39.6%) disease progression for an overall clinical benefit of 60.4%. Median time to progression was 4.0 (range 2-22) months and median overall survival 8.0 (range 3-35) months. Treatment was well tolerated. Of 187 cycles, we did not observe any grade 3/4 toxicity with the exception of a single not-febrile G3 neutropenia. Regardless of severity, main toxic effects observed were nausea in 48.1% and vomiting in 22.9% of patients, anemia in 43.2%, fatigue in 32.6% and leukopenia in 23.2%. CONCLUSION Single-agent oral vinorelbine is extremely safe in elderly patients with advanced NSCLC and ECOG PS of two or more and may represent a valid option in this very special population.
Non-Technical Side of Implementation of Electronic HRM System - Discoursive Exploration of Line Managers' and Employees' Perceptions

Electronic Human Resource Management (e-HRM) is coming to a more full-grown stage within organisational life. Much is assumed and expressed about its advantages, however scientific proof of these advantages is scarce. No clarity exists about the answer to the question whether e-HRM contributes to the effectiveness of HRM processes. This paper contributes to the Enterprise Information Systems field in two ways. Firstly, findings-wise, we present results from the qualitative study on the contribution of e-HRM to HRM effectiveness. The data is collected in a Dutch Ministry of the Interior and Kingdom Relations. Results show that e-HRM applications have some impacts on the HRM practices. However, e-HRM is not perceived by the users as contributing to the HRM effectiveness. Interviews with line managers and employees have revealed interesting differences in their needs and perceptions about functionalities of eHRM applications. Secondly, in this paper we integrate two approaches, namely technology-oriented approach, and organizational processes-oriented approach. An intersection of ITand HRMstudies reveals new possibilities both for scientific and practical implications.
A Prospective Randomized Repeat‐Crossover Comparison of Antitachycardia Pacing With Low‐Energy Cardioversion

BackgroundMultiprogrammable antiarrhythmia devices can treat monomorphic ventricular tachycardia (VT) with autodecremental overdrive pacing and/or with low-energy cardioversion. These two methods provide the opportunity to decrease patient discomfort typically experienced with high-energy pulses. Although both therapies are known to be effective, controversy persists over their relative safety and efficacy Methods and ResultsThe purpose of this study was to examine the safety and efficacy of autodecremental overdrive pacing and low-energy cardioversion in reproducibly terminating monomorphic VT in 24 patients with multiprogrammable antiarrhythmia devices. The protocol required that identical ECG morphology VT be reproducibly induced four times to assess the outcome of antitachycardia pacing and cardioversion twice for each patient in a randomized fashion. Each episode of VT was induced via the implanted device. Autodecremental overdrive pacing initially began with seven stimuli at 97% of the VT cycle length, decrementing by 10 msec per stimulus to a minimum coupling interval of 200 msec. If ineffective, autodecremental overdrive pacing was allowed to iterate three more times for a total of four pacing interventions. With each iteration, one stimulus was added to the pacing train. Similarly, with low-energy cardioversion, up to four therapeutic attempts were made, beginning with a 0.2-J pulse. If ineffective, pulse energy was increased to 0.4, 1.0, and finally 2.0 J. All interventions were automatic without human interference. VT (cycle length, 306±42 msec) was repeatedly terminated in 15 of 24 patients (63%) by autodecremental overdrive pacing and in 18 of 24 patients (75%) by low-energy cardioversion (p=0.53). Eight of the 24 patients (33%) had their VT terminated repeatedly by both therapies. VT accelerated to faster VT or ventricular fibrillation by autodecremental overdrive pacing in four of 24 patients (17%) and by low-energy cardioversion in five of 24 (21%) (p=0.88). Only one of the 24 patients (4%) accelerated with both therapies. No patient was unaffected by either therapy. ConclusionIn the manner programmed, autodecremental overdrive pacing and low-energy cardioversion have similar efficacy and acceleration rates. Response to one therapy does not predict response to the other.
Effects of Hypoxia on Oxidative Capacity of Skeletal Muscle in Trained and Untrained Rats 1

Summary Hypoxia reduces the oxidative capacity of muscle mitochondria in a manner similar to that found with exercise when palmitate, pyruvate and succinate were used as substrates. This suggests that low oxygen tension found in the muscle during exercise may cause the effects observed with exercise; or, hypoxia and exercise cause similar effects because they are mediated through a common process.
Motor Control Strategies Revealed in the Structure of Motor Variability

LATASH, M.L. , J.P. SCHOLZ, and G. SCHÖNER. Motor control strategies revealed in the structure of motor variability. Exerc. Sport Sci. Rev., Vol. 30, No. 1, pp 26–31, 2002. We describe an uncontrolled manifold hypothesis, which suggests a particular solution for the notorious problem of motor redundancy. A body of recent experiments supports the uncontrolled manifold hypothesis and shows its ability to discover biological strategies of the coordination of apparently redundant motor systems. The hypothesis and associated computational apparatus have great potential for application in the areas of motor rehabilitation and motor skill acquisition.
Modelling and validating office information systems: an object and logic oriented approach

Developing information systems for the office environment of today requires powerful representation formalisms and techniques capable of modelling all office elements. Furthermore, these formalisms should provide appropriate facilities for the validation of a conceptual schema. In the paper, it is argued that an office modelling approach should provide semantic account for the various aspects of the schema, as well as facilities for simulating its behaviour. A conceptual modelling language is presented that combines the object oriented and logic programming paradigms, and it is demonstrated how this language can be used to validate the conceptual design of an office information system.
Real-Time Measurements of the Redox States of c-Type Cytochromes in Electroactive Biofilms: A Confocal Resonance Raman Microscopy Study

Confocal Resonance Raman Microscopy (CRRM) was used to probe variations of redox state of c-type cytochromes embedded in living mixed-culture electroactive biofilms exposed to different electrode polarizations, under potentiostatic and potentiodynamic conditions. In the absence of the metabolic substrate acetate, the redox state of cytochromes followed the application of reducing and oxidizing electrode potentials. Real-time monitoring of the redox state of cytochromes during cyclic voltammetry (CV) in a potential window where cytochromes reduction occurs, evidenced a measurable time delay between the oxidation of redox cofactors probed by CV at the electrode interface, and oxidation of distal cytochromes probed by CRRM. This delay was used to tentatively estimate the diffusivity of electrons through the biofilm. In the presence of acetate, the resonance Raman spectra of young (10 days, j = 208±49 µA cm−2) and mature (57 days, j = 267±73 µA cm−2) biofilms show that cytochromes remained oxidized homogeneously even at layers as far as 70 µm from the electrode, implying the existence of slow metabolic kinetics that do not result in the formation of a redox gradient inside the biofilm during anode respiration. However, old biofilms (80 days, j = 190±37 µA cm−2) with thickness above 100 µm were characterized by reduced catalytic activity compared to the previous developing stages. The cytochromes in these biofilm were mainly in the reduced redox state, showing that only aged mixed-culture biofilms accumulate electrons during anode respiration. These results differ substantially from recent observations in pure Geobacter sulfurreducens electroactive biofilms, in which accumulation of reduced cytochromes is already observed in thinner biofilms, thus suggesting different bottlenecks in current production for mixed-culture and G. sulfurreducens biofilms.
Title Tunable diblock copolypeptide hydrogel depots for local delivery of hydrophobic molecules in healthy and injured central nervous system

Many hydrophobic small molecules are available to regulate gene expression and other cellular functions. Locally restricted application of such molecules in the central nervous system (CNS) would be desirable in many experimental and therapeutic settings, but is limited by a lack of innocuous vehicles able to load and easily deliver hydrophobic cargo. Here, we tested the potential for diblock copolypeptide hydrogels (DCH) to serve as such vehicles. In vitro tests on loading and release were conducted with cholesterol and the anti-cancer agent, temozolomide (TMZ). Loading of hydrophobic cargo modified DCH physical properties such as stiffness and viscosity, but these could readily be tuned to desired ranges by modifying DCH concentration, amino acid composition or chain lengths. Different DCH formulations exhibited different loading capacities and different rates of release. For example, comparison of different DCH with increasing alanine contents showed corresponding increases in both cargo loading capacity and time for cargo release. In vivo tests were conducted with tamoxifen, a small synthetic hydrophobic molecule widely used to regulate transgene expression. Tamoxifen released from DCH depots injected into healthy or injured CNS efficiently activated reporter gene expression in a locally restricted manner in transgenic mice. These findings demonstrate the facile and predictable tunability of DCH to achieve a wide range of loading capacities and release profiles of hydrophobic cargos while retaining CNS compatible physical properties. In addition, the findings show that DCH depots injected into the CNS can efficiently deliver small hydrophobic molecules that regulate gene expression in local cells.
Direct measurement of T cell receptor affinity and sequence from naïve antiviral T cells

Direct assessment of TCR affinity and correlated TCR sequence from primary human antigen-specific T cells using iTAST reveals a large range of TCR affinity on HCV precursor T cells that varies with age. iTAST: A taste of T cell specificity There is a well-established link between T cell receptor (TCR) affinity and T cell function, which can have important implications for T cell–based therapies. Zhang et al. have now developed a method to concurrently sequence TCRs and measure TCR affinity of individual T cells from humans. Using this technique, they show that older individuals may be less able to respond to viral infections or vaccines due to a lack of high-affinity T cells. This method could help screen T cells for cancer immunotherapy or vaccine responses. T cells recognize and kill a myriad of pathogen-infected or cancer cells using a diverse set of T cell receptors (TCRs). The affinity of TCR to cognate antigen is of high interest in adoptive T cell transfer immunotherapy and antigen-specific T cell repertoire immune profiling because it is widely known to correlate with downstream T cell responses. We introduce the in situ TCR affinity and sequence test (iTAST) for simultaneous measurement of TCR affinity and sequence from single primary CD8+ T cells in human blood. We demonstrate that the repertoire of primary antigen-specific T cells from pathogen-inexperienced individuals has a surprisingly broad affinity range of 1000-fold composed of diverse TCR sequences. Within this range, samples from older individuals contained a reduced frequency of high-affinity T cells compared to young individuals, demonstrating an age-related effect of T cell attrition that could cause holes in the repertoire. iTAST should enable the rapid selection of high-affinity TCRs ex vivo for adoptive immunotherapy and measurement of T cell response for immune monitoring applications.
Efficacy of a Novel Sublingual Spray Formulation of Artemether in African Children with Plasmodium falciparum Malaria

ABSTRACT The efficacy of sublingual artemether (ArTiMist) was investigated in two studies. In study 1, 31 children were randomized to sublingual artemether (n = 16) or intravenous (i.v.) quinine (n = 15). In study 2, 151 children were randomized to sublingual artemether (n = 77) or i.v. quinine (n = 74). For both studies, patients weighed between 5 and 15 kg and had either severe or complicated malaria based on WHO criteria, or they had uncomplicated malaria but were unable to tolerate oral medication as a result of nausea, vomiting, or diarrhea. Patients received either 3 mg/kg of body weight of sublingual artemether or a loading dose of 20 mg/kg of i.v. quinine followed by 10 mg/kg every 8 h i.v. thereafter. The primary endpoint was parasitological success, defined as a reduction in parasite count of ≥90% of that at baseline at 24 h after the first dose. Other endpoints based on parasite clearance and clinical response were evaluated. In study 1, there were parasitological success rates of 93.3% (14/15) and 66.7% (10/15) for the sublingual artemether and quinine treatments, respectively. In study 2, 94.3% (66/70) of the ArTiMist-treated patients and 39.4% (28/71) of the quinine-treated patients had parasitological success (P < 0.0001). Indicators of parasite clearance (parasite clearance time [PCT], time for parasite count to fall by 50% [PCT50], time for parasite count to fall by 90% [PCT90], and percent reduction in parasitemia from baseline at 24 h [PRR24]) were significantly superior for children treated with sublingual artemether compared to those treated with i.v. quinine. There were no differences between treatments for the clinical endpoints, such as fever clearance time. The local tolerability of sublingual artemether was good. Sublingual artemether leads to rapid parasite clearance and clinical recovery. (Studies 1 and 2 are registered at ClinicalTrials.gov under registration numbers NCT01047436 and NCT01258049, respectively.)
Hemodynamic modifications in patients with symptomatic unilateral stenosis of the internal carotid artery: evaluation with MR imaging perfusion sequences.

BACKGROUND AND PURPOSE Risk of developing ischemia is higher in patients with reduced cerebrovascular reactivity than in those with preserved cerebrovascular reactivity. Therefore, we assessed cerebral hemodynamic modifications in patients with unilateral stenosis of the internal carotid artery by using perfusion-weighted MR imaging to determine if these modifications underlie or anticipate ischemic signs and symptoms. METHODS Fifteen patients with unilateral 70-90% carotid artery stenosis were studied with digital subtraction angiography and perfusion-weighted MR imaging. Their findings were compared with those of 15 age- and sex-matched control subjects. Regional cerebral blood volume (rCBV) and mean transit time (MTT) values were calculated in the middle cerebral artery and border zone territories. RESULTS No significant difference was noted in rCBV and MTT values between the hemispheres in the symptomatic patients. There was a significant difference in MTT values in the border zones between patients and control subjects. MR images in patients and control subjects did not reveal large territorial infarcts and did reveal similar white matter lesion burdens. CONCLUSION There is adequate compensation of unilateral stenosis when the stenosis is less than 90%. The risk of stroke is higher in patients with stenoses exceeding 70%, mostly because of decreased collateral reserve when confronted with emboli.
Smart DNA Fabrication Using Sound Waves

Acoustic droplet ejection (ADE) technology uses focused acoustic energy to transfer nanoliter-scale liquid droplets with high precision and accuracy. This noncontact, tipless, low-volume dispensing technology minimizes the possibility of cross-contamination and potentially reduces the costs of reagents and consumables. To date, acoustic dispensers have mainly been used in screening libraries of compounds. In this paper, we describe the first application of this powerful technology to the rapidly developing field of synthetic biology, for DNA synthesis and assembly at the nanoliter scale using a Labcyte Echo 550 acoustic dispenser. We were able to successfully downscale PCRs and the popular one-pot DNA assembly methods, Golden Gate and Gibson assemblies, from the microliter to the nanoliter scale with high assembly efficiency, which effectively cut the reagent cost by 20- to 100-fold. We envision that acoustic dispensing will become an instrumental technology in synthetic biology, in particular in the era of DNA foundries.