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ACS Nano Nov 2023The application of fluorescent proteins (FPs) in optoelectronics is hindered by the need for effective protocols to stabilize them under device preparation and...
The application of fluorescent proteins (FPs) in optoelectronics is hindered by the need for effective protocols to stabilize them under device preparation and operational conditions. Factors such as high temperatures, irradiation, and organic solvent exposure contribute to the denaturation of FPs, resulting in a low device performance. Herein, we focus on addressing the photoinduced heat generation associated with FP motion and rapid heat transfer. This leads to device temperatures of approximately 65 °C, causing FP-denaturation and a subsequent loss of device functionality. We present a FP stabilization strategy involving the integration of electrostatically self-assembled FP-apoferritin cocrystals within a silicone-based color down-converting filter. Three key achievements characterize this approach: () an engineering strategy to design positively supercharged FPs (+22) without compromising photoluminescence and thermal stability compared to their native form, () a carefully developed crystallization protocol resulting in highly emissive cocrystals that retain the essential photoluminescence features of the FPs, and () a strong reduction of the device's working temperature to 40 °C, leading to a 40-fold increase in Bio-HLEDs stability compared to reference devices.
PubMed: 37902649
DOI: 10.1021/acsnano.3c05284 -
Bioinformation 2023It is of interest to develop potent and safer anti-inflammatory drugs from plants, as medicinal plants and herbs attained great attention in the medical world due to...
It is of interest to develop potent and safer anti-inflammatory drugs from plants, as medicinal plants and herbs attained great attention in the medical world due to their multifunctional activities. This article studied the anti-inflammatory effects of lauric acid (LA), thiocolchicoside (TC) and thiocolchicoside-lauric acid (TC-LA) formulation. The anti-inflammatory effects of these compounds were determined by following the methods of inhibition of protein denaturation and proteinase inhibition activity. This was assessed at different concentrations to determine the 50% inhibition concentration (IC50) of the compounds. The result indicated that the activity of LA, TC, TC-LA formulation, and reference drug increased with the increase in the concentration from 10-50 µg/ml, thus proving the activity of LA, TC, and TC-LA formulation against inflammation was in a dose-dependent manner. The percentage of inhibition of protein denaturation was 59.56%, 66.94%, 86.62%, and 60.34% for LA, TC, the combination of TC-LA and standard drug, and the IC50 values were found to be 44.78 µg/mL, 37.65 µg/mL, 27.15 µg/mL and 43.42 µg/mL, respectively. The percentage of proteinase inhibition activity of LA, TC, and a combination of TC-LA and the standard drug was 66.65%, 77.49%, 94.07%, and 69.83%, and IC50 of LA, TC, a combination of TC-LA and standard drug were35.5 µg/mL, 32.12 µg/mL, 24.35 µg/mL and 37.80 µg/mL, respectively. We found out that lauric acid, thiocolchicoside, and thiocolchicoside-lauric acid formulation exhibited significant anti-inflammatory activity.
PubMed: 38046516
DOI: 10.6026/973206300191075 -
The Journal of Physical Chemistry. B Oct 2023Chemical unfolding with guanidineHCl or urea is a common method to study the conformational stability of proteins. The analysis of unfolding isotherms is usually...
Chemical unfolding with guanidineHCl or urea is a common method to study the conformational stability of proteins. The analysis of unfolding isotherms is usually performed with an empirical linear extrapolation method (LEM). A large positive free energy is assigned to the native protein, which is usually considered to be a minimum of the free energy. The method thus contradicts common expectations. Here, we present a multistate cooperative model that addresses specifically the binding of the denaturant to the protein and the cooperativity of the protein unfolding equilibrium. The model is based on a molecular statistical-mechanical partition function of the ensemble, but simple solutions for the calculation of the binding isotherm and the associated free energy are presented. The model is applied to 23 published unfolding isotherms of small and large proteins. For a given denaturant, the binding constant depends on temperature and pH but shows little protein specificity. Chemical unfolding is less cooperative than thermal unfolding. The cooperativity parameter σ is at least 2 orders of magnitude larger than that of thermal unfolding. The multistate cooperative model predicts zero free energy for the native protein, which becomes strongly negative beyond the midpoint concentration of unfolding. The free energy to unfold a cooperative unit corresponds exactly to the diffusive energy of the denaturant concentration gradient necessary for unfolding. The temperature dependence of unfolding isotherms yields the denaturant-induced unfolding entropy and, in turn, the unfolding enthalpy. The multistate cooperative model provides molecular insight and is as simple to apply as the LEM but avoids the conceptual difficulties of the latter.
PubMed: 37735883
DOI: 10.1021/acs.jpcb.3c03558 -
Frontiers in Allergy 2023Amylase trypsin inhibitors (ATIs) play an important role in wheat allergies and potentially in non-coeliac wheat sensitivity. Food processing could be important to... (Review)
Review
Amylase trypsin inhibitors (ATIs) play an important role in wheat allergies and potentially in non-coeliac wheat sensitivity. Food processing could be important to mitigate the pathogenic properties of ATIs, e.g., by denaturation, glycation, enzymatic hydrolysis, cross-linking, and oxidation and reduction. These modifications also impact the solubility and extractability. The complex solubility behaviour of ATI isoforms (water and salt soluble, but also chloroform-methanol soluble, solubility depending on the redox state) becomes even more complex upon processing due to denaturation and (bio)chemical modifications. This significantly hinders the feasibility of quantitative extraction. Moreover, changes in biofunctionality may occur during the process of extraction, and the changes in ATI due to food processing will be more difficult to assess. Heat treatment decreases the extractability of ATIs with water, NaCl, and other buffer extracts, and binding of IgE from wheat-allergic persons to ATIs as observed with Western blotting is decreased or absent. IgE binding is reduced with the total extract in chaotropic and reducing agents. However, it can be increased when the proteins are hydrolyzed by proteases. Fermentation involving certain species of (FLB), followed by baking, decreases the amount of ATIs and IgE binding to ATIs. In yeast-fermented bread, the amount of ATIs decreased in a similar manner, but IgE binding was more prominent, indicating that there was a modification of ATIs that affected the epitope recognition. When isolated ATIs are ingested with high ATI degrading FLB, the immune response in mice is less elevated , when compared with ATI without high ATI degrading FLB. The pathogenic effects on the skin of dogs and one wheat-allergic child are also decreased when soluble proteins or isolated ATIs are reduced with the thioredoxin/thioredoxin reductase NADPH system. Glycation on the other hand has been shown to potentiate the allergenic properties of ATIs as evidenced by the large increase in IgE binding. The impact of food processing on the pathogenic properties of ATIs is hardly studied in humans. There seem to be opportunities to mitigate the pathogenic properties , but potentiation of pathogenic properties is also frequently observed. This requires a deeper understanding on the impact of food processing on the pathogenicity of ATIs.
PubMed: 38075395
DOI: 10.3389/falgy.2023.1228353 -
ACS Omega Aug 2023and , well known as ginger and lemon, are two vegetals widely used in traditional medicine and the culinary field. The juices of the two vegetals were evaluated based...
and , well known as ginger and lemon, are two vegetals widely used in traditional medicine and the culinary field. The juices of the two vegetals were evaluated based on their inflammation, both in vivo and in vitro. High-performance liquid chromatography (HPLC) was used to characterize different juices from Roscoe and . After the application of the HPLC method, different compounds were identified, such as 6-gingerol and 6-gingediol from the ginger juice and isorhamnetin and hesperidin from the lemon juice. In addition, the two juices and their formulation were assessed for their anti-inflammatory activity, in vitro by utilizing the BSA denaturation test, in vivo using the carrageenan-induced inflammation test, and the vascular permeability test. Important and statistically significant anti-inflammatory activities were observed for all juices, especially the formulation. The results of our work showed clearly that the and juices protect in vivo the development of the rat paw edema, especially the formulation F composed of the and juices, which shows an anti-inflammatory activity equal to -35.95% and -44.05% using 10 and 20 mg/kg of the dose, respectively. Our work also showed that the formulation was the most effective tested extract since it inhibits the vascular permeability by -37% and -44% at the doses of 200 and 400 mg/kg, respectively, and in vitro via the inhibition of the denaturation of BSA by giving a synergetic effect with the highest IC equal to 684.61 ± 7.62 μg/mL corresponding to the formulation F. This work aims to develop nutraceutical preparations in the future and furnishes the support for a new investigation into the activities of the various compounds found in Roscoe and .
PubMed: 37546676
DOI: 10.1021/acsomega.2c04263 -
Circulation. Heart Failure May 2024
PubMed: 38602111
DOI: 10.1161/CIRCHEARTFAILURE.124.011623 -
International Journal of Cosmetic... Oct 2023Skin brightness and spot have a significant impact on youthful and beautiful appearance. One important factor influencing skin brightness is the amount of internal...
Juniper berry extract containing Anthricin and Yatein suppresses lipofuscin accumulation in human epidermal keratinocytes through proteasome activation, increases brightness and decreases spots in human skin.
OBJECTIVE
Skin brightness and spot have a significant impact on youthful and beautiful appearance. One important factor influencing skin brightness is the amount of internal reflected light from the skin. Observers recognize the total surface-reflected light and internal reflected light as skin brightness. The more internal reflected light from the skin, the more attractive and brighter the skin appears. This study aims to identify a new natural cosmetic ingredient that increases the skin's internal reflected light, decreases spot and provides a youthful and beautiful skin appearance.
METHODS
Lipofuscin in epidermal keratinocytes, the aggregating complex of denatured proteins and peroxidized lipids, is one factor that decreases skin brightness and causes of spot. Aggregates block light transmission, and peroxidized lipids lead to skin yellowness, dullness and age spot. Lipofuscin is known to accumulate intracellularly with ageing. Rapid removal of intracellular denatured proteins prevents lipofuscin formation and accumulation in cells. We focused a proteasome system that efficiently removes intracellular denatured proteins. To identify natural ingredients that increase proteasome activity, we screened 380 extracts derived from natural products. The extract with the desired activity was fractionated and purified to identify active compounds that lead to proteasome activation. Finally, the efficacy of the proteasome-activating extract was evaluated in a human clinical study.
RESULTS
We discovered that Juniperus communis fruits (Juniper berry) extract (JBE) increases proteasome activity and suppresses lipofuscin accumulation in human epidermal keratinocytes. We found Anthricin and Yatein, which belong to the lignan family, to be major active compounds responsible for the proteasome-activating effect of JBE. In a human clinical study, an emulsion containing 1% JBE was applied to half of the face twice daily for 4 weeks, resulting in increased internal reflected light, brightness improvement (L-value) and reduction in yellowness (b-value) and spot in the cheek area.
CONCLUSION
This is the first report demonstrating that JBE containing Anthricin and Yatein decreases lipofuscin accumulation in human epidermal keratinocytes through proteasome activation, increases brightness and decreases surface spots in human skin. JBE would be an ideal natural cosmetic ingredient for creating a more youthful and beautiful skin appearance with greater brightness and less spot.
Topics: Humans; Proteasome Endopeptidase Complex; Lipofuscin; Juniperus; Fruit; Keratinocytes; Proteins
PubMed: 37317028
DOI: 10.1111/ics.12876 -
BMC Biomedical Engineering Jul 2023Epilepsy is a neurological disorder that has a variety of origins. It is caused by hyperexcitability and an imbalance between excitation and inhibition, which results in...
BACKGROUND
Epilepsy is a neurological disorder that has a variety of origins. It is caused by hyperexcitability and an imbalance between excitation and inhibition, which results in seizures. The World Health Organization (WHO) and its partners have classified epilepsy as a major public health concern. Over 50 million individuals globally are affected by epilepsy which shows that the patient's family, social, educational, and vocational activities are severely limited if seizures are not controlled. Patients who suffer from epileptic seizures have emotional, behavioral, and neurological issues. Alerting systems using a wearable sensor are commonly used to detect epileptic seizures. However, most of the devices have no multimodal systems that increase sensitivity and lower the false discovery rate for screening and intervention of epileptic seizures. Therefore, the objective of this project was, to design and develop an efficient, economical, and automatically detecting epileptic seizure device in real-time.
METHODS
Our design incorporates different sensors to assess the patient's condition such as an accelerometer, pulsoxymeter and vibration sensor which process body movement, heart rate variability, oxygen denaturation, and jerky movement respectively. The algorithm for real-time detection of epileptic seizures is based on the following: acceleration increases to a higher value of 23.4 m/s or decreases to a lower value of 10 m/s as energy is absorbed by the body, the heart rate increases by 10 bpm from the normal heart rate, oxygen denaturation is below 90% and vibration should be out of the range of 3 Hz -17 Hz. Then, a pulsoxymeter device was used as a gold standard to compare the heart rate variability and oxygen saturation sensor readings. The accuracy of the accelerometer and vibration sensor was also tested by a fast-moving and vibrating normal person's hand.
RESULTS
The prototype was built and subjected to different tests and iterations. The proposed device was tested for accuracy, cost-effectiveness and ease of use. An acceptable accuracy was achieved for the accelerometer, pulsoxymeter, and vibration sensor measurements, and the prototype was built only with a component cost of less than 40 USD excluding design, manufacturing, and other costs. The design is tested to see if it fits the design criteria; the results of the tests reveal that a large portion of the scientific procedures utilized in this study to identify epileptic seizures is effective.
CONCLUSION
This project is objectively targeted to design a medical device with multimodal systems that enable us to accurately detect epileptic seizures by detecting symptoms commonly associated with an episode of epileptic seizure and notifying a caregiver for immediate assistance. The proposed device has a great impact on reducing epileptic seizer mortality, especially in low-resource settings where both expertise and treatment are scarce.
PubMed: 37461102
DOI: 10.1186/s42490-023-00073-7 -
Frontiers in Bioengineering and... 2023Throughout the twenty-first century, the view on inclusion bodies (IBs) has shifted from undesired by-products towards a targeted production strategy for recombinant... (Review)
Review
Throughout the twenty-first century, the view on inclusion bodies (IBs) has shifted from undesired by-products towards a targeted production strategy for recombinant proteins. Inclusion bodies can easily be separated from the crude extract after cell lysis and contain the product in high purity. However, additional solubilization and refolding steps are required in the processing of IBs to recover the native protein. These unit operations remain a highly empirical field of research in which processes are developed on a case-by-case basis using elaborate screening strategies. It has been shown that a reduction in denaturant concentration during protein solubilization can increase the subsequent refolding yield due to the preservation of correctly folded protein structures. Therefore, many novel solubilization techniques have been developed in the pursuit of mild solubilization conditions that avoid total protein denaturation. In this respect, ionic liquids have been investigated as promising agents, being able to solubilize amyloid-like aggregates and stabilize correctly folded protein structures at the same time. This review briefly summarizes the state-of-the-art of mild solubilization of IBs and highlights some challenges that prevent these novel techniques from being yet adopted in industry. We suggest mechanistic models based on the thermodynamics of protein unfolding with the aid of molecular dynamics simulations as a possible approach to solve these challenges in the future.
PubMed: 37545893
DOI: 10.3389/fbioe.2023.1249196 -
Methods in Molecular Biology (Clifton,... 2024In this chapter, we will present the methodology currently applied in our laboratory for the structural elucidation of the cerebrospinal fluid (CSF) N-glycome. N-glycans...
In this chapter, we will present the methodology currently applied in our laboratory for the structural elucidation of the cerebrospinal fluid (CSF) N-glycome. N-glycans are released from denatured carboxymethylated glycoproteins by digestion with peptide-N-glycosidase F (PNGase F) and purified using both C18 Sep-Pak and porous graphitized carbon (PGC) HyperSep™ Hypercarb™ solid phase extraction (SPE) cartridges. The glycan pool is subsequently permethylated to increase mass spectrometry sensitivity. Molecular assignments are performed through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) analysis considering either the protein N-linked glycosylation pathway or MALDI TOF MS/MS data. Each stage has been optimized to obtain high-quality mass spectra in reflector mode with an optimal signal-to-noise ratio up to m/z 4800. This method has been successfully adopted to associate specific N-glycome profiles to the early and the advanced phases of Alzheimer's disease (AD).
Topics: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Glycomics; Tandem Mass Spectrometry; Glycoproteins; Glycosylation; Polysaccharides
PubMed: 38427187
DOI: 10.1007/978-1-0716-3774-6_4