Noise-induced properties of active dendrites.

Dendrites play an essential role in the integration of highly fluctuating input in vivo into neurons across all nervous systems. Yet, they are often studied under conditions where inputs to dendrites are sparse. The dynamic properties of active dendrites facing in vivo-like fluctuating input thus remain elusive. In this paper, we uncover dynamics in a canonical model of a dendritic compartment with active calcium channels, receiving in vivo-like fluctuating input. In a single-compartment model of the active dendrite with fast calcium activation, we show noise-induced nonmonotonic behavior in the relationship of the membrane potential output, and mean input emerges. In contrast, noise can induce bistability in the input-output relation in the system with slowly activating calcium channels. Both phenomena are absent in a noiseless condition. Furthermore, we show that timescales of the emerging stochastic bistable dynamics extend far beyond a deterministic system due to stochastic switching between the solutions. A numerical simulation of a multicompartment model neuron shows that in the presence of in vivo-like synaptic input, the bistability uncovered in our analysis persists. Our results reveal that realistic synaptic input contributes to sustained dendritic nonlinearities, and synaptic noise is a significant component of dendritic input integration.

Excitable dynamics in a molecularly-explicit model of cell motility: Mixed-mode oscillations and beyond.

Mesenchymal cell motility is mainly regulated by two members of the Rho-family of GTPases, called Rac and Rho. The mutual inhibition exerted by these two proteins on each other's activation and the promotion of Rac activation by an adaptor protein called paxillin have been implicated in driving cellular polarization comprised of front (high active Rac) and back (high active Rho) during cell migration. Mathematical modeling of this regulatory network has previously shown that bistability is responsible for generating a spatiotemporal pattern underscoring cellular polarity called wave-pinning when diffusion is included. We previously developed a 6V reaction-diffusion model of this network to decipher the role of Rac, Rho and paxillin (along with other auxiliary proteins) in generating wave-pinning. In this study, we simplify this model through a series of steps into an excitable 3V ODE model comprised of one fast variable (the scaled concentration of active Rac), one slow variable (the maximum paxillin phosphorylation rate - turned into a variable) and a very slow variable (a recovery rate - also turned into a variable). We then explore, through slow-fast analysis, how excitability is manifested by showing that the model can exhibit relaxation oscillations (ROs) as well as mixed-mode oscillations (MMOs) whose underlying dynamics are consistent with a delayed Hopf bifurcation with a canard explosion. By reintroducing diffusion and the scaled concentration of inactive Rac into the model, we obtain a 4V PDE model that generates several unique spatiotemporal patterns that are relevant to cell motility. These patterns are then characterized and their impact on cell motility are explored by employing the cellular potts model (CPM). Our results reveal that wave pinning produces purely very directed motion in CPM, while MMOs allow for meandering and non-motile behaviors to occur. This highlights the role of MMOs as a potential mechanism for mesenchymal cell motility.

Ultra-high-performance liquid chromatography using a fused-core particle column for fast analysis of propolis phenolic compounds.

Propolis is a bee product with a complex chemical composition formed by several species from different geographical origins. The complex propolis composition requires an accurate and reproducible characterization of samples to standardize the quality of the material sold to consumers. This work developed an ultra-high-performance liquid chromatography with a photodiode array detector method to analyze propolis phenolic compounds based on the two key propolis biomarkers, Artepillin C and p-Coumaric acid. This choice was made due to the complexity of the sample with the presence of several compounds. The optimized method was hyphenated with mass spectrometry detection allowing the detection of 23 different compounds. A step-by-step strategy was used to optimize temperature, flow rate, mobile phase composition, and re-equilibration time. Reverse-phase separation was achieved with a C18 fused-core column packed with the commercially available smallest particles (1.3 nm). Using a fused-core column with ultra-high-performance liquid chromatography allows highly efficient, sensitive, accurate, and reproducible determination of compounds extracted from propolis with an outstanding sample throughput and resolution. Optimized conditions permitted the separation of the compounds in 5.50 min with a total analysis time (sample-to-sample) of 6.50 min.

Real-Time Detection of Phenylacetaldehyde in Wine: Application of a Microwave Sensor Based on Molecularly Imprinted Silica.

Molecularly imprinted sol-gel silica (MIS) coupled to a microwave sensor was designed and used to detect phenylacetaldehyde (PAA), a chemical tracer of wine oxidation. The developed method is fast, cheap and could replace the classical chromatographic methods, which require a tedious sample preparation and are expensive. To reach our objective, five MIS and their control non-imprinted silica (NIS) were synthesized and their extraction capacity toward PAA was studied in hydro alcoholic medium. The selected polymers, based on this first step, were subjected to a selectivity study in the presence of PAA and three other competing molecules. The best polymer was integrated in a microwave sensor and was used to assess PAA in red wine. The developed sensor was able to detect PAA at the µg·L-1 level, which is below the off-flavour threshold.

Deciphering the dynamics of lamellipodium in a fish keratocytes model.

Motile cells depend on an intricate network of feedback loops that are essential in driving cell movement. Integrin-based focal adhesions (FAs) along with actin are the two key factors that mediate such motile behaviour. Together, they generate excitable dynamics that are essential for forming protrusions at the leading edge of the cell and, in certain cases, traveling waves along the membrane. A partial differential equation (PDE) model of a self-organizing lamellipodium in crawling keratocytes has been previously developed to understand how the three spatiotemporal patterns of activity observed in such cells, namely, stalling, waving and smooth motility, are produced. The model consisted of three key variables: the density of barbed actin filaments, newly formed FAs called nascent adhesions (NAs) and VASP, an anti-capping protein that gets sequestered by NAs during maturation. Using parameter sweeping techniques, the distinct regimes of behaviour associated with the three activity patterns were identified. In this study, we convert the PDE model into an ordinary differential equation (ODE) model to examine its excitability properties and determine all the patterns of activity exhibited by this system. Our results reveal that there are two additional regimes not previously identified, including bistability and oscillatory-like type IV excitability (generated by three steady states and their manifolds, rather than limit cycles). These regimes are also present in the PDE model. Applying slow-fast analysis on the ODE model shows that it exhibits a canard explosion through a folded-saddle and that rough motility seen in keratocytes is likely due to noise-dependent motility governed by dynamics near the interface of bistability and type IV excitability. The two parameter bifurcation suggests that the increase in the proportion of rough motion is due to a shift in activity towards the bistable and type IV excitable regimes induced by a decrease in NA maturation rate. Our results thus provide important insight into how microscopic mechanical effects are integrated to produce the observed modes of motility.

Rapid method for simultaneous determination of ascorbic acid and zinc in effervescent tablets by capillary zone electrophoresis with contactless conductivity detection.

Ascorbic acid and zinc are essential nutrients that play important roles in nutrition, immune support, and maintenance of health. For this reason, both compounds are widely used as ingredients in dietary supplements. We report, for the first time, an analytical method for fast simultaneous determination of ascorbic acid and zinc. A single analysis run is possible every 80 s (45 injections/h). The method is based on capillary electrophoresis with capacitively coupled contactless conductivity detection using a fused silica capillary with 50 cm length (effective length of 10 cm). The separation was achieved by using a background electrolyte composed by 30 mmol/L of 2-(morpholin-4-yl)ethane-1-sulfonic acid and 30 mmol/L of histidine, pH 6.1. The detection limits were 10 and 20 µmol/L and recovery values for spiked samples were 101 and 100% for zinc and ascorbic acid, respectively. The results obtained with the developed procedure were compared to those obtained by titration (ascorbic acid) and flame atomic absorption spectroscopy (zinc), and no statistically significant differences were observed (95% confidence level).

Fast determination of codeine, orphenadrine, promethazine, scopolamine, tramadol, and paracetamol in pharmaceutical formulations by capillary electrophoresis.

Paracetamol is an active ingredient commonly found in pharmaceutical formulations in combination with one of the following compounds: codeine, orphenadrine, promethazine, scopolamine, and tramadol. In this work, we propose a unique analytical method for determination of these active ingredients in pharmaceutical samples. The method is based on capillary electrophoresis with capacitively coupled contactless conductivity detection. The separation was achieved on a fused silica capillary (50 cm total length, 40 cm effective length, and 50 µm id) using an optimized background electrolyte composed of 20 mmol/L ß-alanine/4 mmol/L sodium chloride/4 µmol/L sodium hydroxide (pH 9.6). Each sample can be analyzed in a single run (≤2 min) and the limits of detection were 2.5, 0.62, 0.63, 2.5, 15, and 1.6 µmol/L for scopolamine, tramadol, orphenadrine, promethazine, codeine, and paracetamol, respectively. Recovery values for spiked samples were between 94 and 104%.

Simultaneous determination of halogenated contaminants and polycyclic aromatic hydrocarbons: a multi-analyte method applied to filter-feeding edible organisms.

This study develops and validates a novel analytical approach for the simultaneous determination of 127 polychlorinated biphenyls (PCBs), together with 6 polychlorinated naphthalenes (PCNs) and 16 polycyclic aromatic hydrocarbons (PAHs). PCBs, PCNs, and PAHs were subjected to a unique pretreatment protocol and were simultaneously determined in a single chromatographic run, using GC-MS, in environmental marine samples of mussels and clams. The results of the validation experiments, which were performed on the standard reference materials (NIST SRM 1974C - slurried matrix and NIST SRM 2977 - freeze-dried matrix), were in accordance with the certified and the reference values. The repeatability of the method for all target compounds, expressed as mean relative standard deviations, ranged from 2.5 to 5.1 % for PCBs, from 3.9 to 5.5 % for PCNs, and from 8.6 to 17.9 % for PAHs; the first value of each pair refers to the freeze-dried matrix and the second to the fresh one, for each of the classes of compounds examined. The quantification limits were in the range of 0.2-6 pg for PCBs, 0.4-8 pg for PCNs, and 0.2-15 pg for PAHs (on column). The method recoveries yielded good results (62 ± 19 % for the freeze-dried matrix and 60 ± 14 % for the fresh one) and were not significantly reduced by adopting a single analytical protocol compared with the use of different group-specific analytical methods. No serious interferences were encountered and good selectivity was achieved. These results show that this method allows one to increase the laboratory sample throughput while requiring a small amount of tissue and saving time. Graphical Abstract Simultaneous determination of halogenated contaminants and polycyclic aromatic hydrocarbons: a multianalyte method applied to filter-feeding edible organisms.

Basin of Attraction of Solutions with Pattern Formation in Slow-Fast Reaction-Diffusion Systems.

This article is devoted to the characterization of the basin of attraction of pattern solutions for some slow-fast reaction-diffusion systems with a symmetric property and an underlying oscillatory reaction part. We characterize some subsets of initial conditions that prevent the dynamical system to evolve asymptotically toward solutions which are homogeneous in space. We also perform numerical simulations that illustrate theoretical results and give rise to symmetric and non-symmetric pattern solutions. We obtain these last solutions by choosing particular random initial conditions.

C18-bound porous silica monolith particles as a low-cost high-performance liquid chromatography stationary phase with an excellent chromatographic performance.

Ground porous silica monolith particles with an average particle size of 2.34 µm and large pores (363 Å) exhibiting excellent chromatographic performance have been synthesized on a relatively large scale by a sophisticated sol-gel procedure. The particle size distribution was rather broad, and the d(0.1)/d(0.9) ratio was 0.14. The resultant silica monolith particles were chemically modified with chlorodimethyloctadecylsilane and end-capped with a mixture of hexamethyldisilazane and chlorotrimethylsilane. Very good separation efficiency (185,000/m) and chromatographic resolution were achieved when the C18 -bound phase was evaluated for a test mixture of five benzene derivatives after packing in a stainless-steel column (1.0 mm × 150 mm). The optimized elution conditions were found to be 70:30 v/v acetonitrile/water with 0.1% trifluoroacetic acid at a flow rate of 25 µL/min. The column was also evaluated for fast analysis at a flow rate of 100 µL/min, and all the five analytes were eluted within 3.5 min with reasonable efficiency (ca. 60,000/m) and resolution. The strategy of using particles with reduced particle size and large pores (363 Å) combined with C18 modification in addition to partial-monolithic architecture has resulted in a useful stationary phase (C18 -bound silica monolith particles) of low production cost showing excellent chromatographic performance.

Determination of flavor enhancers in milk powder by one-step sample preparation and two-dimensional liquid chromatography.

Maltol, ethyl maltol, vanillin, and ethyl vanillin are important food additives as flavor enhancers. To quantify the four additives in milk powder, a novel 2D liquid chromatographic (2DLC) method was developed in this article. In such a 2DLC system, the target fractions eluted from the first dimensional column (C4) are stored onto the trapping column (C8) for subsequent analysis; after that, they were switched into the second dimensional column (C18) by a two-position six-port switching valve. A one-step sample preparation method was used prior to 2DLC chromatographic analysis, which was easy and convenient. After optimization of all experimental parameters, the new method was validated in terms of linearity, LODs, and LOQs, intra- and interday precision, and accuracy. A conventional single-dimensional liquid chromatographic method was also proposed in this work for comparison. In order to evaluate the applicability of the new 2DLC method, five brands of commercial milk powder samples (n = 8) were analyzed. Vanillin and ethyl vanillin were detected in two samples, respectively. It is showed that the 2DLC method is effective in quality control programs of milk powder products.

Solid-phase extraction with the metal-organic framework MIL-101(Cr) combined with direct analysis in real time mass spectrometry for the fast analysis of triazine herbicides.

MIL-101(Cr) is an excellent metal-organic framework with high surface area and nanoscale cavities, making it promising in solid-phase extraction. Herein, we used MIL-101(Cr) as a solid-phase extraction packing material combined with fast detection of direct analysis in real time mass spectrometry (DART-MS) for the analysis of triazine herbicides. After systematic optimization of the operation parameters, including the gas temperature of DART, the moving speed of the 1D platform, solvent for desorption, amount of MIL-101(Cr) extraction time, eluent volume and salt concentration, this method can realize the simultaneous detection of five kinds of triazine herbicides. The limits of detection were 0.1∼0.2 ng/mL and the linear ranges covered more than two orders of magnitude with the quantitation limits of 0.5∼1 ng/mL. Moreover, the developed method has been applied for the analysis of lake water samples and the recoveries for spiked analytes were in the range of 85∼110%. These results showed that solid-phase extraction with metal-organic frameworks is an efficient sample preparation approach for DART-MS analysis and could find more applications in environmental analysis.

A Robust Parallel Computing Data Extraction Framework for Nanopore Experiments.

The success of a nanopore experiment relies not only on the quality of the experimental design but also on the performance of the analysis program utilized to decipher the ionic perturbations necessary for understanding the fundamental molecular intricacies. An event extraction framework is developed that leverages parallel computing, efficient memory management, and vectorization, yielding significant performance enhancement. The newly developed abf-ultra-simple function extracts key parameters from the header critical for the operation of open-seek-read-close data loading architecture running on multiple cores. This underpins the swift analysis of large files where an ≈ × 18 improvement is found for a 100 min-long file (≈4.5 GB) compared to the more traditional single (cell) array data loading method. The application is benchmarked against five other analysis platforms showcasing significant performance enhancement (>2 ×-1120 ×). The integrated provisions for batch analysis enable concurrently analyzing multiple files (vital for high-bandwidth experiments). Furthermore, the application is equipped with multi-level data fitting based on abrupt changes in the event waveform. The application condenses the extracted events to a single binary file improving data portability (e.g., 16 GB file with 28 182 events reduces to 47.9 MB-343 × size reduction) and enables a multitude of post-analysis extractions to be done efficiently.

Improved chiral SFC screening for analytical method development.

In this study we describe the evaluation of a recently developed supercritical fluid chromatography (SFC) instrument for automated chiral SFC method development. The greatly improved gradient dwell volume and liquid flow control of the new instrument in combination with the use of shorter columns containing smaller stationary phase particles affords chiral SFC method development that is faster and more universal than previous systems.

Ultra-short columns for the chromatographic analysis of large molecules.

Today, reverse phase liquid chromatography (RPLC) analysis of proteins is almost exclusively performed on conventional columns (100-150 mm) in gradient elution mode. However, it was shown many years ago that large molecules present an on/off retention mechanism, and that only a very short inlet segment of the chromatographic column retains effectively the large molecules. Much shorter columns - like only a few centimetres or even a few millimetres - can therefore be used to efficiently analyse such macromolecules. The aim of this review is to summarise the historical and more recent works related to the use of very short columns for the analysis of model and therapeutic proteins. To this end, we have outlined the theoretical concepts behind the use of short columns, as well as the instrumental limitations and potential applications. Finally, we have shown that these very short columns were also possibly interesting for other chromatographic modes, such as ion exchange chromatography (IEX), hydrophilic interaction chromatography (HILIC) or hydrophobic interaction chromatography (HIC), as analyses in these chromatographic modes are performed in gradient elution mode.

Combining micro- and portable-XRF as a tool for fast identification of virus infections in plants: The case study of ASa-Virus in Fraxinus ornus L.

Plant viruses can affect micro- and macro-nutrients homeostasis in woody plants, with fluctuation in the concentration of some elements at the leaf level due to the pathogen activity and/or the plant physiological response to the infection.Leaves of Fraxinus ornus L. (flowering ash) were sampled for three consecutive years in the city of Hamburg (Germany), from both trees showing the typical symptoms of the ash shoestring associated virus infection (ASaV+) and healthy trees (ASaV-). Such leaves were analyzed by µ-XRF, using both laboratory and synchrotron X-ray sources, and large differences between symptomatic and not symptomatic leaves were observed: ASaV+ samples showed uneven element distribution and regions of the lamina with severe depletions of P, S, and Ca. Differently, K appeared more concentrated. Thus, 139 leaflets sampled from various healthy and infected ash trees over the three-year period were analyzed for K and Ca concentration with a portable XRF instrument. We found that the K:Ca concentration ratio was always significantly higher in ASaV+ samples, and this trend was verified for all the samplings over the tree years. We conclude that the K:Ca ratio parameter has potential in the frame of trendsetting diagnostics and could be used, together with visual symptoms, for a rapid, non-destructive, on-site and cheap indirect ASaV detection.

Rapid analysis to distinguish porcine and bovine gelatin using PANI/NiO nanoparticles modified Quartz Crystal Microbalance (QCM) sensor.

Rapid analysis to distinguish porcine and bovine gelatin using a modified Quartz Crystal Microbalance (QCM) sensor has been studied. The PANI was deposited on the sensor surface using electropolymerization, and then nickel nanoparticles were deposited by layer by layer (LbL) technique. The modified QCM sensor's performance was compared to an unmodified sensor in porcine and bovine gelatin at neutral, acidic, and alkaline conditions. The result shows that the unmodified sensor cannot distinguish between porcine and bovine gelatin, whereas the modified QCM sensor produces a different response. Porcine gelatin shows an increasing frequency response, but in contrast, bovine gelatin decreases frequency response at the alkaline condition. The time response was 2 min with a detection limit of 51.2 ppm and 8.7 ppm for porcine and bovine gelatin, respectively. Further investigation shows that the modified sensor can analyze porcine gelatin contamination in the a mixed gelatin sample.

Fluorescence spectrophotometry for COVID-19 determination in clinical swab samples.

Considering the limitations of the assays currently available for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its emerging variants, a simple and rapid method using fluorescence spectrophotometry was developed to detect coronavirus disease 2019 (COVID-19). Forty clinical swab samples were collected from the nasopharyngeal and oropharyngeal cavities of COVID-19-positive and -negative. Each sample was divided into two parts. The first part of the samples was analyzed using reverse transcription-polymerase chain reaction (RT-qPCR) as the control method to identify COVID-19-positive and -negative samples. The second part of the samples was analyzed using fluorescence spectrophotometry. Fluorescence measurements were performed at excitation and emission wavelengths ranging from 200 to 800 nm. Twenty COVID-19-positive samples and twenty COVID-19-negative samples were detected based on RT-qPCR results. The fluorescence spectrum data indicated that the COVID-19-positive and -negative samples had significantly different characteristics. All positive samples could be distinguished from negative samples by fluorescence spectrophotometry. Principal component analysis showed that COVID-19-positive samples were clustered separately from COVID-19-negative samples. The specificity and accuracy of this experiment reached 100%. Limit of detection (LOD) obtained 42.20 copies/ml (Ct value of 33.65 cycles) for E gene and 63.60 copies/ml (Ct value of 31.36 cycles) for ORF1ab gene. This identification process only required 4 min. Thus, this technique offers an efficient and accurate method to identify an individual with active SARS-CoV-2 infection and can be easily adapted for the early investigation of COVID-19, in general.

Development of Narrow-Bore C18 Column for Fast Separation of Peptides and Proteins in High-Performance Liquid Chromatography.

Separation with high efficiency and good resolution is constantly in demand in the pharmaceutical industry. The fast and efficient separation of complex samples such as peptides and proteins is a challenging task. To achieve high efficiency with good resolution, chromatographers are moving towards small particles packed into narrow-bore columns. Silica monolith particles (sub-2 µm) were derivatized with chlorodimethyl octadecyl silane (C18) and packed into stainless steel columns (100 mm × 1.8 mm i.d) by a slurry-packing method. The developed columns were used for the separation of peptides and proteins. A separation efficiency (N) of 40,000 plates/column (400,000 plates/m) was achieved for the mixture of five peptides. Similarly, the fast separation of the peptides was carried out using a high flow rate, and the separation of the five peptides was achieved in one minute with high efficiency (N ≅ 240,000 plates/m). The limit of detection (DL) and the limit of quantification (QL) for each analyte were determined by developing a linear regression curve with relatively very low concentrations of the target compound. The average values of the QL for the peptide and proteins were 0.55 ng and 0.48 ng, respectively, using short C18 column (1.8 mm × 100 mm) UV (at 214 nm). The fast analysis of peptides and proteins with such high efficiency and good resolution has not been reported in the literature yet. Owing to high efficiency, these home-made columns could be used as an alternative to the expensive commercial columns for peptide and protein separation.

Ion Chromatography-High-Resolution Mass Spectrometry Method for the Determination of Bromide Ions in Cereals and Legumes: New Scenario for Global Food Security.

The new scenario for global food production and supply is decidedly complex given the current forecast of an increase in food fragility due to international tensions. In this period, exports from other parts of the world require different routes and treatments to preserve the food quality and integrity. Fumigation is a procedure used for the killing, removal, or rendering infertile of pests, with serious dangers to human health. The most-used fumigants are methyl bromide and ethylene dibromide. It is important to bear in mind that the soil may contain bromide ions naturally or from anthropogenic source (fertilizers and pesticides that contain bromide or previous fumigations). Different methods (titrimetric, spectrophotometric, and fluorometric approaches) are available to rapidly determine the amount of bromide ion on site in the containers, but these are non-specific and with high limits of quantification. The increasing interest in healthy food, without xenobiotic residues, requires the use of more sensitive, specific, and accurate analytical methods. In order to help give an overview of the bromide ion scenario, a new, fast method was developed and validated according to SANTE 11312/2021. It involves the determination of bromide ion in cereals and legumes through ion chromatography-Q-Orbitrap. The extraction was performed by the QuPPe method, but some modifications were applied based on the matrix. The method described here was validated at four different levels. Recoveries were satisfactory and the mean values ranged between 99 and 106%, with a relative standard deviation lower than 3%. The linearity in the matrix was evaluated to be between 0.010 and 2.5 mg kg-1, with a coefficient of determination (R2) of 0.9962. Finally, the proposed method was applied to different cereals and legumes (rice, wheat, beans, lentils pearled barley, and spelt) and tested with satisfactory results in EUPT-SMR16 organized by EURL.