Unbinding Transition Induced by Osmotic Pressure in Relation to Unilamellar Vesicle Formation

Small-angle X-ray scattering and phase-contrast microscopy experiments were performed to investigate the effect of the osmotic pressure on vesicle formation in a dioleoylphosphatidylcholine (DOPC)/water/NaI system. Multi-lamellar vesicles were formed when a pure lipid film was hydrated with an aqueous solution of NaI. On the other hand, uni-lamellar vesicles (ULVs) were formed when a lipid film mixed with an enough amount of NaI was hydrated. To confirm the effect of the osmotic pressure due to NaI, a free-energy calculation was performed. This result showed that the osmotic pressure induced an unbinding transition on the hydration process, which resulted in ULV formation

HD Physiology Project-Japanese efforts to promote multilevel integrative systems biology and physiome research.

The HD Physiology Project is a Japanese research consortium that aimed to develop methods and a computational platform in which physiological and pathological information can be described in high-level definitions across multiple scales of time and size. During the 5 years of this project, an appropriate software platform for multilevel functional simulation was developed and a whole-heart model including pharmacokinetics for the assessment of the proarrhythmic risk of drugs was developed. In this article, we outline the description and scientific strategy of this project and present the achievements and influence on multilevel integrative systems biology and physiome research

Facilitation of I Kr current by some hERG channel blockers suppresses early afterdepolarizations.

Drug-induced block of the cardiac rapid delayed rectifying potassium current (I Kr), carried by the human ether-a-go-go-related gene (hERG) channel, is the most common cause of acquired long QT syndrome. Indeed, some, but not all, drugs that block hERG channels cause fatal cardiac arrhythmias. However, there is no clear method to distinguish between drugs that cause deadly arrhythmias and those that are clinically safe. Here we propose a mechanism that could explain why certain clinically used hERG blockers are less proarrhythmic than others. We demonstrate that several drugs that block hERG channels, but have favorable cardiac safety profiles, also evoke another effect; they facilitate the hERG current amplitude in response to low-voltage depolarization. To investigate how hERG facilitation impacts cardiac safety, we develop computational models of I Kr block with and without this facilitation. We constrain the models using data from voltage clamp recordings of hERG block and facilitation by nifekalant, a safe class III antiarrhythmic agent. Human ventricular action potential simulations demonstrate the ability of nifekalant to suppress ectopic excitations, with or without facilitation. Without facilitation, excessive I Kr block evokes early afterdepolarizations, which cause lethal arrhythmias. When facilitation is introduced, early afterdepolarizations are prevented at the same degree of block. Facilitation appears to prevent early afterdepolarizations by increasing I Kr during the repolarization phase of action potentials. We empirically test this prediction in isolated rabbit ventricular myocytes and find that action potential prolongation with nifekalant is less likely to induce early afterdepolarization than action potential prolongation with dofetilide, a hERG channel blocker that does not induce facilitation. Our data suggest that hERG channel blockers that induce facilitation increase the repolarization reserve of cardiac myocytes, rendering them less likely to trigger lethal ventricular arrhythmias

Difference in trafficking of brain-derived neurotrophic factor between axons and dendrites of cortical neurons, revealed by live-cell imaging

BACKGROUND: Brain-derived neurotrophic factor (BDNF), which is sorted into a regulated secretory pathway of neurons, is supposed to act retrogradely through dendrites on presynaptic neurons or anterogradely through axons on postsynaptic neurons. Depending on which is the case, the pattern and direction of trafficking of BDNF in dendrites and axons are expected to be different. To address this issue, we analyzed movements of green fluorescent protein (GFP)-tagged BDNF in axons and dendrites of living cortical neurons by time-lapse imaging. In part of the experiments, the expression of BDNF tagged with cyan fluorescent protein (CFP) was compared with that of nerve growth factor (NGF) tagged with yellow fluorescent protein (YFP), to see whether fluorescent protein-tagged BDNF is expressed in a manner specific to this neurotrophin. RESULTS: We found that BDNF tagged with GFP or CFP was expressed in a punctated manner in dendrites and axons in about two-thirds of neurons into which plasmid cDNAs had been injected, while NGF tagged with GFP or YFP was diffusely expressed even in dendrites in about 70% of the plasmid-injected neurons. In neurons in which BDNF-GFP was expressed as vesicular puncta in axons, 59 and 23% of the puncta were moving rapidly in the anterograde and retrograde directions, respectively. On the other hand, 64% of BDNF-GFP puncta in dendrites did not move at all or fluttered back and forth within a short distance. The rest of the puncta in dendrites were moving relatively smoothly in either direction, but their mean velocity of transport, 0.47 ± 0.23 (SD) μm/s, was slower than that of the moving puncta in axons (0.73 ± 0.26 μm/s). CONCLUSION: The present results show that the pattern and velocity of the trafficking of fluorescence protein-tagged BDNF are different between axons and dendrites, and suggest that the anterograde transport in axons may be the dominant stream of BDNF to release sites

Selective Pruning of More Active Afferents When Cat Visual Cortex Is Pharmacologically Inhibited

AbstractActivity-dependent competition is thought to guide the normal development of specific patterns of neural connections. Such competition generally favors more active inputs, making them larger and stronger, while less active inputs become smaller and weaker. We pharmacologically inhibited the activity of visual cortical cells and measured the three-dimensional structure of inputs serving the two eyes when one eye was occluded. The more active inputs serving the open eye actually became smaller than the deprived inputs from the occluded eye, which were similar to those in normal animals. These findings demonstrate in vivo that it is not the amount of afferent activity but the correlation between cortical and afferent activity that regulates the growth or retraction of these inputs

All-or-none switching of transcriptional activity on single DNA molecules caused by a discrete conformational transition

Recently, it has been confirmed that long duplex DNA molecules with sizes larger than several tens of kilo-base pairs (kbp), exhibit a discrete conformational transition from an elongated coil state to a compact globule state upon the addition of various kinds of chemical species that usually induce DNA condensation. In this study, we performed a single-molecule observation on a large DNA, Lambda ZAP II DNA (ca. 41 kbp), in a solution containing RNA polymerase and substrates along with spermine, a tetravalent cation, at different concentrations, by use of fluorescence staining of both DNA and RNA. We found that transcription, or RNA production, is completely inhibited in the compact state, but is actively performed in the unfolded coil state. Such an all-or-none effect on transcriptional activity induced by the discrete conformational transition of single DNA molecules is discussed in relation to the mechanism of the regulation of large-scale genetic activity.Comment: 14 pages, 2 figure

A Proposal of a Privacy-preserving Questionnaire by Non-deterministic Information and Its Analysis

We focus on a questionnaire consisting of three-choice question or multiple-choice question, and propose a privacy-preserving questionnaire by non-deterministic information. Each respondent usually answers one choice from the multiple choices, and each choice is stored as a tuple in a table data. The organizer of this questionnaire analyzes the table data set, and obtains rules and the tendency. If this table data set contains personal information, the organizer needs to employ the analytical procedures with the privacy-preserving functionality. In this paper, we propose a new framework that each respondent intentionally answers non-deterministic information instead of deterministic information. For example, he answers ‘either A, B, or C’ instead of the actual choice A, and he intentionally dilutes his choice. This may be the similar concept on the k-anonymity. Non-deterministic information will be desirable for preserving each respondent\u27s information. We follow the framework of Rough Non-deterministic Information Analysis (RNIA), and apply RNIA to the privacy-preserving questionnaire by non-deterministic information. In the current data mining algorithms, the tuples with non-deterministic information may be removed based on the data cleaning process. However, RNIA can handle such tuples as well as the tuples with deterministic information. By using RNIA, we can consider new types of privacy-preserving questionnaire.2016 IEEE International Conference on Big Data, December 5-8, 2016, Washington DC, US

Effects of Syringe Material and Silicone Oil Lubrication on the Stability of Pharmaceutical Proteins

ABSTRACTCurrently, polymer-based prefillable syringes are being promoted to the pharmaceutical market because they provide an increased break resistance relative to traditionally used glass syringes. Despite this significant advantage, the possibility that barrel material can affect the oligomeric state of the protein drug exists. The present study was designed to compare the effect of different syringe materials and silicone oil lubrication on the protein aggregation. The stability of a recombinant fusion protein, abatacept (Orencia), and a fully human recombinant immunoglobulin G1, adalimumab (Humira), was assessed in silicone oil-free (SOF) and silicone oil-lubricated 1-mL glass syringes and polymer-based syringes in accelerated stress study. Samples were subjected to agitation stress, and soluble aggregate levels were evaluated by size-exclusion chromatography and verified with analytical ultracentrifugation. In accordance with current regulatory expectations, the amounts of subvisible particles resulting from agitation stress were estimated using resonant mass measurement and dynamic flow-imaging analyses. The amount of aggregated protein and particle counts were similar between unlubricated polymer-based and glass syringes. The most significant protein loss was observed for lubricated glass syringes. These results suggest that newly developed SOF polymer-based syringes are capable of providing biopharmaceuticals with enhanced physical stability upon shipping and handling. © 2015 The Authors. Journal of Pharmaceutical Sciences published by Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:527–535, 201

Predicting inpatient length of stay in a Portuguese hospital using the CRISP-DM methodology

Com base nos dados disponíveis num hospital português relativos aos processos de internamento, ocorridos no período de 2000 a 2013, e seguindo a metodologia de data mining CRISP-DM, obteve-se um modelo de previsão dos tempos de internamento baseado no algoritmo random forest que apresentou uma elevada qualidade, e superior à obtida com outras técnicas de data mining, e que permitiu identificar os atributos clínicos do paciente como os mais importantes para a explicação dos tempos de internamento.Using data collected from a Portuguese hospital, within the period 2000 to 2013, we adopted the CRISP-DM methodology to predict inpatient length of stay. The best method (random forest algorithm) achieved a high quality prediction. Such model allowed the identification of the most relevant input features, which are related with the patients’ clinical attributes.(undefined