Identification of calcium chelating peptides from peanut protein hydrolysate and absorption activity of peptide-calcium complex.

BACKGROUND: Peanut peptides have good chelating ability with metal ions. However, there are few studies on the chelation mechanism of peanut peptides with calcium and absorption properties of peptide-calcium complex. RESULTS: Peptides with high calcium chelating rate were isolated and purified from peanut protein hydrolysate (PPH), and the chelation rate of component F21 was higher (81.4 ± 0.8%). Six peptides were identified from component F21 by liquid chromatography-tandem mass spectrometry, and the frequency of acidic amino acids and arginine in the amino acid sequence was higher in all six peptides. Peanut peptide-calcium complex (PPH21-Ca) was prepared by selecting component F21 (PPH21). Ultraviolet analysis indicated that the chelate reaction occurred between peanut peptide and calcium ions. Fourier transform infrared analysis showed that the chelating sites were carboxyl and amino groups on the amino acid residues of peptides. Scanning electron microscopy revealed that the surface of peanut peptide had a smooth block structure, but the surface of the complex had a granular morphology. Caco-2 cell model tests revealed that the bioavailability of PPH21-Ca was 58.4 ± 0.5%, which was significantly higher than that of inorganic calcium at 37.0 ± 0.4%. CONCLUSION: Peanut peptides can chelate calcium ions by carboxyl and amino groups, and the peptide-calcium complex had higher bioavailability. This study provides a theoretical basis for the development of new calcium supplement products that are absorbed easily. © 2024 Society of Chemical Industry.

Evaluation of oilseed proteins as precursors of antimicrobial peptides using bioinformatics method.

In this study, the potential of oilseed proteins from soybean, peanut, sesame, sunflower seed and flaxseed as antimicrobial peptide (AMP) precursors was assessed using the bioinformatics method. Thirty-four novel potential AMPs were obtained by in silico hydrolysis of 12 oilseed protein sequences, and 11 of them were positive in all four algorithm tests in CAMPR3. Among the six proteases analyzed, trypsin cleaved soybean, peanut, sesame and sunflower seed proteins most effectively to generate AMPs, with three, four, two and two AMPs obtained, respectively. Subtilisin was most effective for flaxseed AMPs release, obtaining three AMPs. More than 85% of AMPs were predicted to be cationic peptides, and some AMPs were hydrophobic. These potential AMPs were classified as non-toxic peptides, and 15 peptides were non-allergenic. All the AMPs were unstable to digestive enzymes according to in silico simulated digestion. The results of this study provide a theoretical basis for further development of AMPs using oilseed proteins.

Changes in, and correlation analysis of, volatile compounds, key enzymes, and fatty acids in lemon juice vesicles during freeze drying and hot-air drying.

BACKGROUND: Lemon juice vesicles are distinguished by their unique and abundant volatile flavor compounds, which can undergo complex changes during drying. In this study, integrated freeze drying (IFD), conventional freeze drying (CFD), and hot-air drying (AD) were used to dry lemon juice vesicles to investigate the changes in, and correlations among volatile compounds, fatty acids, and key enzyme activity during the drying process. RESULTS: Twenty-two volatile compounds were detected during the drying processes. Compared with fresh samples, seven compounds were lost in the dried samples after IFD, seven after CFS, and six after AD, and the loss rates of the total content of volatile compounds in the dried samples were 82.73% in CFD, more than 71.22% in IFD, and more than 28.78% in AD. In total, 1.015 mg/g of seven fatty acids were detected in the fresh samples; the content loss rates of total fatty acids after drying were 67.68% in AD, more than 53.00% in CFD, and more than 36.95% in IFD, respectively. During the three drying processes, IFD retained relatively higher enzyme activity in the samples. CONCLUSION: Many positive and negative correlations (P < 0.05) were observed among the key enzyme effects, fatty acids, and volatile compounds, showing close associations. The current work provides information that is important for the selection of suitable drying techniques for lemon juice vesicles and suggests how to control their flavor during the drying process. © 2023 Society of Chemical Industry.

Dynamic Hand Gesture Recognition Using Electrical Impedance Tomography.

Electrical impedance tomography (EIT) has been applied in the field of human-computer interaction due to its advantages including the fact that it is non-invasive and has both low power consumption and a low cost. Previous work has focused on static gesture recognition based on EIT. Compared with static gestures, dynamic gestures are more informative and can achieve more functions in human-machine collaboration. In order to verify the feasibility of dynamic gesture recognition based on EIT, a traditional excitation drive pattern is optimized in this paper. The drive pattern of the fixed excitation electrode is tested for the first time to simplify the measurement process of the dynamic gesture. To improve the recognition accuracy of the dynamic gestures, a dual-channel feature extraction network combining a convolutional neural network (CNN) and gated recurrent unit (GRU), namely CG-SVM, is proposed. The new center distance loss is designed in order to simultaneously supervise the intra-class distance and inter-class distance. As a result, the discriminability of the confusing data is improved. With the new excitation drive pattern and classification network, the recognition accuracy of different interference data has increased by 2.7~14.2%. The new method has stronger robustness, and realizes the dynamic gesture recognition based on EIT for the first time.

Hydrophilic, Clean Graphene for Cell Culture and Cryo-EM Imaging.

The wettability of graphene is critical for numerous applications but is very sensitive to its surface cleanness. Herein, by clarifying the impact of intrinsic contamination, i.e., amorphous carbon, which is formed on the graphene surface during the high-temperature chemical vapor deposition (CVD) process, the hydrophilic nature of clean graphene grown on single-crystal Cu(111) substrate was confirmed by both experimental and theoretical studies, with an average water contact angle of ∼23°. Furthermore, the wettability of as-transferred graphene was proven to be highly dependent on its intrinsic cleanness, because of which the hydrophilic, clean graphene exhibited improved performance when utilized for cell culture and cryoelectron microscopy imaging. This work not only validates the intrinsic hydrophilic nature of graphene but also provides a new insight in developing advanced bioapplications using CVD-grown clean graphene films.

Three-dimensional measurement method of color fringe projection based on an improved three-step phase-shifting method.

A three-dimensional (3D) measurement method of color fringe projection based on an improved three-step phase-shifting method is proposed. The color fringe pattern is encoded by two cosine fringe patterns with the same frequency but different shifting phase and a uniform gray flat image into three color channels R, G, and B. Although the measurement speed of the traditional three-step phase-shifting method can meet the requirements of measuring 3D objects, it makes the noise and inaccuracy of the captured images increase, and each image will cause measurement error. Therefore, we improve the three-step phase-shifting method and introduce the Hilbert transform into the three-step phase-shift method. The DC component of the fringe pattern is obtained by using the Hilbert transform principle, and the third fringe pattern in the three-step phase-shift method is replaced by the captured light intensity distribution of the DC component. The phase difference of the other two fringe patterns is fixed as π/2 by the Hilbert transform. The improved three-step phase-shifting method is used to obtain the phase information of the deformed color fringe image, and then the phase-unwrapping algorithm is used to obtain the phase distribution information of the whole field. The results show that the improved method can not only accurately calculate the phase information but also greatly improve the measurement speed and quality.

Development of a revised ICC-qPCR method used for Pseudorabies virus inactivation validation study of biologically sourced materials.

To develop a precise and convenient method to evaluate the virus transmission risk of biologically sourced materials, an integrated cell culture-qPCR (ICC-qPCR) method for Pseudorabies virus (PRV) was established and revised for applications to this new field. The optimized post-infection period was found at 12-hr to achieve a reasonable detection limit (-0.25 Log10TCID50/100 µL, Logs) and a quantitative range (0.75-3.75 Logs). The results of mimic samples suggested that three 10-fold dilutions at the time of virus inoculation combined with three washes after virus absorption, and the sets of non-amplified samples as controls could efficiently eliminate the false positive signals caused by high levels of noninfectious viruses. The virus inactivation validation studies of acellular porcine corneas suggested that the logs inactivation of PRV at 12 kGy irradiation dose obtained by general ICC-qPCR, revised ICC-qPCR and cell culture were 2.49, 4.85 and 5.08, respectively. At 25 kGy, those were 2.31, 4.85 and 5.08, respectively. The results obtained by the revised ICC-qPCR were consistent with cell culture and more precise than general ICC-qPCR. Therefore, the revised ICC-qPCR proposed in this study has an application prospect in the PRV inactivation validation studies of biologically sourced materials.

Soft and MRI Compatible Neural Electrodes from Carbon Nanotube Fibers.

Soft and magnetic resonance imaging (MRI) compatible neural electrodes enable stable chronic electrophysiological measurements and anatomical or functional MRI studies of the entire brain without electrode interference with MRI images. These properties are important for many studies, ranging from a fundamental neurophysiological study of functional MRI signals to a chronic neuromodulatory effect investigation of therapeutic deep brain stimulation. Here we develop soft and MRI compatible neural electrodes using carbon nanotube (CNT) fibers with a diameter from 20 µm down to 5 µm. The CNT fiber electrodes demonstrate excellent interfacial electrochemical properties and greatly reduced MRI artifacts than PtIr electrodes under a 7.0 T MRI scanner. With a shuttle-assisted implantation strategy, we show that the soft CNT fiber electrodes can precisely target specific brain regions and record high-quality single-unit neural signals. Significantly, they are capable of continuously detecting and isolating single neuronal units from rats for up to 4-5 months without electrode repositioning, with greatly reduced brain inflammatory responses as compared to their stiff metal counterparts. In addition, we show that due to their high tensile strength, the CNT fiber electrodes can be retracted controllably postinsertion, which provides an effective and convenient way to do multidepth recording or potentially selecting cells with particular response properties. The chronic recording stability and MRI compatibility, together with their small size, provide the CNT fiber electrodes unique research capabilities for both basic and applied neuroscience studies.

Stretchable Transparent Electrode Arrays for Simultaneous Electrical and Optical Interrogation of Neural Circuits in Vivo.

Recent developments of transparent electrode arrays provide a unique capability for simultaneous optical and electrical interrogation of neural circuits in the brain. However, none of these electrode arrays possess the stretchability highly desired for interfacing with mechanically active neural systems, such as the brain under injury, the spinal cord, and the peripheral nervous system (PNS). Here, we report a stretchable transparent electrode array from carbon nanotube (CNT) web-like thin films that retains excellent electrochemical performance and broad-band optical transparency under stretching and is highly durable under cyclic stretching deformation. We show that the CNT electrodes record well-defined neuronal response signals with negligible light-induced artifacts from cortical surfaces under optogenetic stimulation. Simultaneous two-photon calcium imaging through the transparent CNT electrodes from cortical surfaces of GCaMP-expressing mice with epilepsy shows individual activated neurons in brain regions from which the concurrent electrical recording is taken, thus providing complementary cellular information in addition to the high-temporal-resolution electrical recording. Notably, the studies on rats show that the CNT electrodes remain operational during and after brain contusion that involves the rapid deformation of both the electrode array and brain tissue. This enables real-time, continuous electrophysiological monitoring of cortical activity under traumatic brain injury. These results highlight the potential application of the stretchable transparent CNT electrode arrays in combining electrical and optical modalities to study neural circuits, especially under mechanically active conditions, which could potentially provide important new insights into the local circuit dynamics of the spinal cord and PNS as well as the mechanism underlying traumatic injuries of the nervous system.

[Single-use Medical Devices Re-processing: Risk Assessment and Quality Control Technologies].

The reuse of high-cost single-use medical devices (SUD) is permitted in many countries, such as the United States, Germany and the United Kingdom, but strict regulatory requirements must be met. In addition to regulatory policies and regulations, such as market access mode and special requirements on Good Manufacture Practice (GMP), there are strict technical requirements on the potential risk control and quality assurance system. Therefore, effective risk assessment and risk control technology are the keys to ensure effective quality control and safe use of SUDs. In this article, based on analyzing the technological requirements of the national regulatory on SUDs in the United States, Germany and Britain, and combined with the review from latest relevant literature, to discuss the strategies of how to carry out scientific risk assessment. Some risk control technologies on the reuse of SUDs are introduced, which will provide support for the further study on risk control strategies and regulatory decisions for the reuse of SUDs in China.

[Single-use Medical Devices Re-processing: Regulatory Status Quo].

Some single-use medical devices are reprocessed and reused in some countries in the world, but the regulatory approach is different, and in some countries it isn't regulated yet. In this article, the regulatory status quo of single-use medical devices is reviewed. The regulatory development, important regulatory documents and regulatory approaches of single-use medical device reprocessing in the United States, Germany and the UK are introduced. And how to perform scientific risk assessment and effective risk control is discussed. The information is useful to establish China-specific regulations, and to develop relevant standards, guidelines or specifications and the risk control strategies.

[Bioresorbable Vascular Scaffold: A Focused Review on Development and Preclinical Studies].

Bioresorbable vascular scaffolds(BVS) are new treatment strategies of percutaneous coronary intervention. They have been introduced to overcome limitations of bare metal stents (BMS) and drug-eluting stents(DES), since they provide temporary scaffolding and then disappear, liberate the treated vessel from cage. In this article, we review the current status and problems of BVS, various tests required before gaining regulatory approval for clinical use.

[Evaluation of Material Permeability of Type I Collagen Hydrogel].

OBJECTIVES: To establish an experimental method for evaluating material permeability of type I collagen hydrogels. METHODS: Using BSA-FITC as an indicator, by combining BSA-FITC with PBS they were used as permeability media, and using transwell load hydrogen sample to detect BSA-FITC transparent rate. RESULTS: In the concentration range of 100 µg·mL-1~0.781 µg·mL-1, the standard curve R2 ≥ 0.99, Lower Limit of Quantity (LLOQ) is 3.125 µg·mL-1, RSD <5%, detection recovery rate is in the range of 80%~120%. CONCLUSIONS: In this study, we established an experimental method for evaluating material permeability of hydrogel. The BSA-FITC transparent rate of type I collagen hydrogel was 100% at 28 h.

Sub-10-nm intracellular bioelectronic probes from nanowire-nanotube heterostructures.

The miniaturization of bioelectronic intracellular probes with a wide dynamic frequency range can open up opportunities to study biological structures inaccessible by existing methods in a minimally invasive manner. Here, we report the design, fabrication, and demonstration of intracellular bioelectronic devices with probe sizes less than 10 nm. The devices are based on a nanowire-nanotube heterostructure in which a nanowire field-effect transistor detector is synthetically integrated with a nanotube cellular probe. Sub-10-nm nanotube probes were realized by a two-step selective etching approach that reduces the diameter of the nanotube free-end while maintaining a larger diameter at the nanowire detector necessary for mechanical strength and electrical sensitivity. Quasi-static water-gate measurements demonstrated selective device response to solution inside the nanotube, and pulsed measurements together with numerical simulations confirmed the capability to record fast electrophysiological signals. Systematic studies of the probe bandwidth in different ionic concentration solutions revealed the underlying mechanism governing the time response. In addition, the bandwidth effect of phospholipid coatings, which are important for intracellular recording, was investigated and modeled. The robustness of these sub-10-nm bioelectronics probes for intracellular interrogation was verified by optical imaging and recording the transmembrane resting potential of HL-1 cells. These ultrasmall bioelectronic probes enable direct detection of cellular electrical activity with highest spatial resolution achieved to date, and with further integration into larger chip arrays could provide a unique platform for ultra-high-resolution mapping of activity in neural networks and other systems.

Graphene Encapsulated Copper Microwires as Highly MRI Compatible Neural Electrodes.

Magnetic resonance imaging (MRI) compatible neural electrodes are important for combining high-resolution electrophysiological measurements with more global MRI mapping of brain activity, which is critical for fundamental neuroscience studies, as well as clinical evaluation and monitoring. Copper is a favorable material to use in MRI because it has magnetic susceptibility close to water and tissues. However, the cytotoxicity of copper precludes its direct implantation for neural recording. Here, we overcome this limitation by developing a graphene encapsulated copper (G-Cu) microelectrode. The toxicity of copper is largely eliminated, as evidenced by the in vitro cell tests and in vivo histology studies. Local field potentials and single-unit spikes were recorded from rodent brains with the G-Cu microelectrodes. Notably, the G-Cu microelectrodes show no image artifacts in a 7.0 T MRI scanner, indicating minimal magnetic field distortion in their vicinity. This high MRI compatibility of our G-Cu probes would open up new opportunities for fundamental brain activity studies and clinical applications requiring continuous MRI and electrophysiological recordings.

High-level expression and biochemical characterization of a novel cold-active lipase from Rhizomucor endophyticus.

OBJECTIVES: To identify novel cold-active lipases from fungal sources and improve their production by heterologous expression in Pichia pastoris. RESULTS: A novel cold-active lipase gene (ReLipB) from Rhizomucor endophyticus was cloned. ReLipB was expressed at a high level in Pichia pastoris using high cell-density fermentation in a 5-l fermentor with the highest lipase activity of 1395 U/ml. The recombinant lipase (RelipB) was purified and biochemically characterized. ReLipB was most active at pH 7.5 and 25 °C. It was stable from pH 4.5-9.0. It exhibited broad substrate specificity towards p-nitrophenyl (pNP) esters (C2-C16) and triacylglycerols (C2-C12), showing the highest specific activities towards pNP laurate (231 U/mg) and tricaprylin (1840 U/mg), respectively. In addition, the enzyme displayed excellent stability with high concentrations of organic solvents including cyclohexane, n-hexane, n-heptane, isooctane and petroleum ester and surfactants. CONCLUSIONS: A novel cold-active lipase from Rhizomucor endophyticus was identified, expressed at a high level and biochemically characterized. The high yield and unique enzymatic properties make this lipase of some potential for industrial applications.

Structural insights into the substrate specificity of two esterases from the thermophilic Rhizomucor miehei.

Two hormone-sensitive lipase (HSL) family esterases (RmEstA and RmEstB) from the thermophilic fungus Rhizomucor miehei, exhibiting distinct substrate specificity, have been recently reported to show great potential in industrial applications. In this study, the crystal structures of RmEstA and RmEstB were determined at 2.15 Å and 2.43 Å resolutions, respectively. The structures of RmEstA and RmEstB showed two distinctive domains, a catalytic domain and a cap domain, with the classical α/ß-hydrolase fold. Catalytic triads consisting of residues Ser161, Asp262, and His292 in RmEstA, and Ser164, Asp261, and His291 in RmEstB were found in the respective canonical positions. Structural comparison of RmEstA and RmEstB revealed that their distinct substrate specificity might be attributed to their different substrate-binding pockets. The aromatic amino acids Phe222 and Trp92, located in the center of the substrate-binding pocket of RmEstB, blocked this pocket, thus narrowing its catalytic range for substrates (C2-C8). Two mutants (F222A and W92F in RmEstB) showing higher catalytic activity toward long-chain substrates further confirmed the hypothesized interference. This is the first report of HSL family esterase structures from filamentous fungi. The information on structure-function relationships could open important avenues of exploration for further industrial applications of esterases.

Sinusoidal phase-modulating fiber-optic interferometer fringe with a feedback control system.

A displacement measurement system using a fiber-optic interferometer fringe projector with a feedback control system is presented and demonstrated. The system utilizes the integrating bucket method to detect the desired phase or the displacement and Fresnel reflection signal to realize measurement of the disturbance and feed it back to the modulated signal of the laser at the same time. The continuous signal truly reflects the error information, as the output light and reflected light share the same optical path. Practical experiments validate the feasibility of this method.

Changes and correlation analysis of volatile flavor compounds, amino acids, and soluble sugars in durian during different drying processes.

Durian contains rich flavor components that undergo complex changes during drying. In this study, durian was subjected to integrated freeze-drying (IFD), conventional freeze-drying (CFD), and hot air drying (AD). Compared with the fresh samples, those dried by IFD, CFD, and AD lost 11, 9, and 7 original volatile compounds, respectively, and generated 7, 6, and 8 new volatile compounds, respectively, and showed a rapid and then slow decreasing trend in the total content during drying. However, the types of amino acids and soluble sugars remained unchanged during each of the drying methods. Furthermore, volatile compounds showed a significant negative correlation with the majority of amino acids and a significant positive correlation with soluble sugars. The IFD samples had the highest content of volatile compounds, amino acids, and soluble sugars. Therefore, IFD is recommended as a preferable drying method for durian.

A high-density 1,024-channel probe for brain-wide recordings in non-human primates.

Large-scale neural population recordings with single-cell resolution across the primate brain remain challenging. Here we introduce the Neuroscroll probe that isolates single neuronal activities simultaneously from 1,024 densely spaced channels that are flexibly distributed across the shank of the probe. The Neuroscroll probe length is easily tunable for individual probes from 10 mm to 90 mm, covering the brain size of non-human primates and humans, and the probes remain intact and functional after repeated bending deformations. The Neuroscroll probes provided reliable recordings from large neural populations with high chronic stability up to 105 weeks in rats. Recording with each Neuroscroll probe yielded hundreds of well-isolated single units simultaneously from multiple brain regions distributed across the entire depth of the rhesus macaque brain. With the thousand simultaneously recorded channels, unprecedented probe length, excellent mechanical stability and flexible recording site distribution, the Neuroscroll probes enable a wide range of new experimental paradigms in system neuroscience studies with great versatility.