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There is current interest in replacing petroleum-based additives in consumer paper products with abundantly available, renewable and sustainable biopolymers such as lignin-containing cellulose nanofibers (LCNFs) and cottonseed protein. This research characterized the performance of cottonseed protein isolate with/without LCNFs to increase the dry strength of filter paper. The application of 10% protein solution with 2% LCNFs as an additive improved the elongation at break, tensile strength and modulus of treated paper products compared to the improved performance of cottonseed protein alone. Improvements in tensile modulus and tensile strength were greatest for samples containing larger amounts of lignin and a greater degree of polymerization than for those with less lignin from the same biomass sources. [ABSTRACT FROM AUTHOR]

Background: Autohydrolysis is an extensively investigated pretreatment method due to its environmental friendliness. During autohydrolysis, most xylan from hemicellulose can be converted into xylooligosaccharides (XOS), and cellulose in the autohydrolyzed residues can be transformed into glucose after enzymatic hydrolysis. Both of these are value-added biochemicals in the biorefining process. In this work, paper mulberry (PM), which contains abundant protein, was utilized as a raw material to coproduce XOS and single-cell protein (SCP) through autohydrolysis and fermentation technologies. Results: The results showed that 8.3 g of XOS and 1.8 g of amino acids could be recovered in the autohydrolysate (based on 100 g raw material) after autohydrolysis (170 °C, 1 h). Moreover, 5.7 g of low-DP XOS along with 1.8 g of amino acids could be further obtained from the autohydrolysate after hydrolysis with endo-β-1-4-xylanase. In addition, 20.1 g of fermentable monosaccharides was recovered after hydrolyzing the autohydrolyzed PM with cellulase, which can be used to produce 4.8 g of SCP after fermentation with Candida utilis. Conclusion: As a valuable application of PM, a novel process is proposed to coproduce amino acid-rich XOS and SCP through autohydrolysis. The carbohydrate of PM is effectively converted to high value-added products. [ABSTRACT FROM AUTHOR]

Exposure to natural rubber latex (NRL) can result in sensitivity to NRL protein with resulting allergic reactions. Low-cost, portable, simple, sensitive analytical tools for NRL protein measurements are needed for rapid and accurate assessments of allergenic risks at the point-of-care (POC) instead of using traditional methods that require large and expensive instruments, long-time analysis, and complex sample preparation steps. Here, an electrochemical paper-based analytical device (ePAD) is presented by combining sample preparation and electrochemical detection within a single device to offer a one-step NRL analysis. The lack of antibodies and/or enzymes against NRL makes POC analysis difficult. In this work, detection is based on electrochemical measurement of the remaining Cu after in situ protein complexation instead of more complex biological assays for the first time. Graphene screen-printed electrodes modified with 1,10-phenanthroline and Nafion were used in the ePAD to improve Cu signal 18-fold relative to unmodified carbon screen-printed electrodes. The optimum parameters including 1,10-phenanthroline concentration, reaction time between Cu and protein, and the starting Cu concentration were 5% w/v, 1 min, and 600 μg mL−1, respectively. In addition to short analysis time (4 min), the system selectivity indicated no other interfering species affecting protein detection. The proposed ePAD achieved an LOD of 3.0 mg dL−1 and a linear range of 10.0–200.0 mg dL−1. Finally, the proposed sensors were applied for NRL protein detection and the results were not significantly different from the traditional Lowry method at 95% confidence. [ABSTRACT FROM AUTHOR]

Paper-based analytical device (PAD) has received more and more attention in the field of point-of-care test (POCT) due to its low cost, portability and simple operation. Sensitivity and selectivity are two important aspects in clinical diagnostic analysis. However, low sensitivity of a PAD limits its wider application for POCT. Here we introduced a PAD that can clean and enrich the protein content from salty media with both electric field (E) and pH gradient (double E/pH gradients), with which 100-fold enrichment effect could be achieved within 70 s as demonstrated by fluorescein isothiocyanate labeled bovine serum albumin (FITC-BSA) from artificial urine media. With post staining of the protein stacking band with bromophenol blue (BPB), selective colorimetric detection of human serum albumin (HSA) was achieved simply with smartphone camera in the clinically significant range of 10–300 mg‧L−1 (R2 = 0.99) with a limit of detection (LOD) of 4.9 mg‧L−1. Detection of microalbuminuria (MAU) of diabetic patients was demonstrated with this method without difference (ɑ = 0.01) to that by the clinical method (immunoturbidimetry). This work demonstrated the potential of this PAD method in online sample pretreatment and detection of target component from complex physiological samples. Image 1 • Both electric field and pH gradient (E/pH double gradients) can be established and sustained on a paper fluidic channel. • Ampholytic content from salty sample media can be stacked/separated from other types of species. • Proteins from urine can be cumulatively stacked on the double gradient interface without intensive pre-treatment. • Sensitive and selective colorimetric detection of microalbuminuria (MAU) from clinical samples was successfully demonstrated. [ABSTRACT FROM AUTHOR]

Proteinaceous toxins are harmful proteins derived from plants, bacteria, and other natural sources. They pose a risk to human health due to infection and also as possible biological warfare agents. Paper spray mass spectrometry (PS-MS) with wipe sampling was used to detect proteins from surfaces as a potential tool for identifying the presence of these toxins. Proteins ranging in mass between 12.4 and 66.5 kDa were tested, including a biological toxin simulant/vaccine for Staphylococcal enterotoxin B (SEBv). Various substrates were tested for these representative proteins, including a laboratory bench, a notebook cover, steel, glass, plant leaf and vinyl flooring. Carbon sputtered porous polyethylene (CSPP) was found to outperform typical chromatography paper used for paper spray, as well as carbon nanotube (CNT)-coated paper and polyethylene (PE), which have been previously shown to be well-suited for protein analysis. Low microgram quantities of the protein toxin simulant and other test proteins were successfully detected with good signal-to-noise from surfaces using a porous wipe. These applications demonstrate that PS-MS can potentially be used for rapid, sample preparation-free detection of proteins and biological warfare agents, which would be beneficial to first responders and warfighters. [ABSTRACT FROM AUTHOR]

This work presents an annotation tool that automatically locates mentions of particular amino‐acid residues in published papers and identifies the protein concerned. These matches can be provided in context or in a searchable format in order for researchers to better use the existing and future literature. [ABSTRACT FROM AUTHOR]

This work reports on fully integrated "sample‐in‐signal‐out" microfluidic paper‐based analytical devices (μPADs) relying on bioluminescence resonance energy transfer (BRET) switches for analyte recognition and colorimetric signal generation. The devices use BRET‐based antibody sensing proteins integrated into vertically assembled layers of functionalized paper, and their design enables sample volume‐independent and fully reagent‐free operation, including on‐device blood plasma separation. User operation is limited to the application of a single drop (20–30 μL) of sample (serum, whole blood) and the acquisition of a photograph 20 min after sample introduction, with no requirement for precise pipetting, liquid handling, or analytical equipment except for a camera. Simultaneous detection of three different antibodies (anti‐HIV1, anti‐HA, and anti‐DEN1) in whole blood was achieved. Given its simplicity, this type of device is ideally suited for user‐friendly point‐of‐care testing in low‐resource environments. A single drop of blood on paper: Bioluminescent sensing proteins integrated into a multi‐layer paper‐based device allow the user‐friendly and simple quantification of monoclonal antibodies from a single drop of blood by simple color change monitoring. [ABSTRACT FROM AUTHOR]

Abstract A novel affinity paper-based electrochemical impedance device (PEID) was first fully developed for cardiovascular risk assessment. Herein, a simple folding PEID comprising a dual screen-printed electrode was fabricated for label-free electrochemical impedance detection. The results demonstrated in a step-wise manner that the phosphocholine-modified screen-printed carbon electrodes were highly responsive to the clinically required range of C-reactive protein (CRP) (0.005 – 500 mg L−1; r2 = 0.993) levels with a detection limit (3σ/slope) of 0.001 mg L−1. The optimal binding frequency of CRP–phosphocholine interaction was determined to be 100 Hz. These results implied that our proposed system could be used for simultaneously measuring the CRP levels using a single PEID platform in combination with the specific recognition elements. When assaying two levels of CRP, the overall assay reproducibility for each concentration, expressed as relative standard error of the mean (RSE%; n = 30), was 1.21%. The variation in the measurements between individual electrodes, expressed as the relative standard deviation (RSD), was 2.82%. Using 2 measurement sites per device, the proposed sensor provided excellent precision for the simultaneous detection of CRP. Moreover, the RSD for the CRP levels measured on ten independently fabricated paper-based sheets was 2.11%, thereby offering an acceptable fabrication reproducibility. The presented folding PEIDs were used forthe determination of CRP in patient-derived blood samples with minimised bias and excellent correlation with a standard method (n = 15; CUSUM linearity test, p > 0.10), thus permitting the potential application of PEID for assessing cardiovascular risk in individuals. Highlights • A PEID was developed for ultrasensitive label-free EIS detection of hs-CRP. • A simply folding step intended to create a small chamber for electrochemical assay. • A single specific-frequency determination facilitates fast impedance measurements. • An excellent agreement was demonstrated between the PEID and the standard method. [ABSTRACT FROM AUTHOR]

Biosensors have emerged as a valuable tool with high specificity and sensitivity for fast and reliable detection of hazardous substances in drinking water. Numerous substances have been addressed using synthetic biology approaches. However, many proposed biosensors are based on living, genetically modified organisms and are therefore limited in shelf life, usability and biosafety. We addressed these issues by the construction of an extensible, cell-free biosensor. Storage is possible through freeze drying on paper. Following the addition of an aqueous sample, a highly efficient cell-free protein synthesis (CFPS) reaction is initiated. Specific allosteric transcription factors modulate the expression of ‘superfolder’ green fluorescent protein (sfGFP) depending on the presence of the substance of interest. The resulting fluorescence intensities are analyzed with a conventional smartphone accompanied by simple and cheap light filters. An ordinary differential equitation (ODE) model of the biosensors was developed, which enabled prediction and optimization of performance. With an optimized cell-free biosensor based on the Shigella flexneri MerR transcriptional activator, detection of 6 μg/L Hg(II) ions in water was achieved. Furthermore, a completely new biosensor for the detection of gamma-hydroxybutyrate (GHB), a substance used as date-rape drug, was established by employing the naturally occurring transcriptional repressor BlcR from Agrobacterium tumefaciens. [ABSTRACT FROM AUTHOR]

There have been limited studies on the potential use of enzymes for enhancing the dewaterability of biosludge. The mechanisms for such enhancement have not been investigated despite the environmental advantages of using enzymes over synthetic polymers for biosludge conditioning. In order to find enzymes with this potential, a screening of commercially available enzymes was carried out using capillary suction time to assess biosludge dewaterability. The only enzyme that showed dewatering improvements in the screening tests was a lysozyme which reduced the capillary suction time by 36% and increased the cake solids content from 5.6 to 8.9 DS%. Lysozyme aided in the flocculation of particles reducing the polymer demand from 11% to 6%. Active and inactive lysozyme exhibited a similar ability for enhancing sludge dewatering, indicating that the conditioning mechanism of lysozyme is similar to that of a flocculant. [ABSTRACT FROM AUTHOR]

Background: It is important for industries to find green chemistries for manufacturing their products that have utility, are cost-effective and that protect the environment. The paper industry is no exception. Renewable resources derived from plant components could be an excellent substitute for the chemicals that are currently used as paper binders. Air laid pressed paper products that are typically used in wet wipes must be bound together so they can resist mechanical tearing during storage and use. The binders must be strong but cost-effective. Although chemical binders are approved by the Environmental Protection Agency, the public is demanding products with lower carbon footprints and that are derived from renewable sources. Results: In this project, carbohydrates, proteins and phenolic compounds were applied to air laid, pressed paper products in order to identify potential renewable green binders that are as strong as the current commercial binders, while being organic and renewable. Each potential green binder was applied to several filter paper strips and tested for strength in the direction perpendicular to the cellulose fibril orientation. Out of the twenty binders surveyed, soy protein, gelatin, zein protein, pectin and Salix lignin provided comparable strength results to a currently employed chemical binder. Conclusions: These organic and renewable binders can be purchased in large quantities at low cost, require minimal reaction time and do not form viscous solutions that would clog sprayers, characteristics that make them attractive to the non-woven paper industry. As with any new process, a large-scale trial must be conducted along with an economic analysis of the procedure. However, because multiple examples of "green" binders were found that showed strong cross-linking activity, a candidate for commercial application will likely be found. [ABSTRACT FROM AUTHOR]

Abstract In this study, levels of inflammatory protein biomarkers in venous plasma, plasma derived from capillary blood from the earlobe, and capillary plasma stored as dried plasma spots (DPS) were compared. Samples from 12 male individuals were assessed with a panel of 92 inflammation-related proteins using multiplex proximity extension assay. Correlations between sample types varied greatly between analytes. A high correlation of ρ > 0.8 was observed between capillary plasma and DPS for 32 analytes. At this level of correlation, 13 analytes correlated between venous and capillary plasma and 5 analytes in the comparison of venous blood with DPS. Highlights • Measurement of an inflammation protein panel in three sample sources. • Earlobe plasma as a less invasive alternative sample source than venous plasma. • Protein measurement in capillary plasma stored as dried plasma spots (DPS). • Correlation value of ρ > 0.8 between capillary earlobe plasma and DPS for 32 analytes. • Correlation value of ρ > 0.8 between venous and capillary earlobe plasma for 13 analytes. [ABSTRACT FROM AUTHOR]

Sensitivity is an important factor determining successful mass spectrometry (MS) analysis of metabolome, protein, drugs, and environmental samples. Currently, nanoelectrospray ionization (ESI) is widely used as a sensitive ionization method. However, application of nano-ESI is limited to polar molecules and there is no atmospheric pressure ionization technique developed that can be used for MS analysis of low- and nonpolar compounds with sensitivity that can match with nano-ESI. Herein, we propose paper spray chemical ionization (PSCI) as an ionization technique that can be used to analyze low- and nonpolar aromatic compounds with high sensitivity. PSCI is based on paper spray ionization utilizing corona discharge phenomenon. PSCI can sensitively and quantitatively detect down to picogram (or femtomole) levels of low- and nonpolar aromatic compounds. [ABSTRACT FROM AUTHOR]

The article focuses on the prognostic significance of galectin-3 in patients with ST-elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI). Topics include the association between high galectin-3 levels and heart failure development, rehospitalization, and adverse cardiovascular events in STEMI patients post-PCI, though it's not an independent predictor of cardiovascular mortality.

Type 1 diabetes (T1D) is increasing in incidence and predictable with measurement of serum islet autoantibodies (iAb) years prior to clinical disease onset. Identifying iAb positive individuals reduces diabetic ketoacidosis and identifies individuals for T1D prevention trials. However, large scale screening for iAb remains challenging as assays have varying sensitivities and specificities, insulin autoantibodies remain difficult to measure and venipuncture is generally required to obtain serum. We developed an approach to reliably measure all four major iAb, including insulin autoantibodies, from dried blood spots (DBS) on filter-paper. By spiking iAb positive serum into iAb negative whole blood in a dose titration, we optimized the conditions for autoantibody elution from filter paper as measured by fluid phase radioimmunoassays. After assessing stability of measuring iAb from DBS over time, we then screened iAb from DBS and the corresponding serum in new-onset T1D (n = 52), and controls (n = 72) which included first-degree relatives of T1D patients. iAb measured from eluted DBS in new-onset T1D strongly correlated with serum measurements (R2 = 0.96 for mIAA, GADA = 0.94, IA-2A = 0.85, ZnT8A = 0.82, p<0.01 for each autoantibody). There were no false positives in control subjects, and 5/6 with previously unknown iAb positivity in sera were detected using DBS. With further validation, measuring iAb from DBS can be a reliable method to screen for T1D risk. [ABSTRACT FROM AUTHOR]

Due to the unstable chemical properties and poor water solubility of carotenoids, their processing adaptation and oral bioavailability are poor, limiting their application in hydrophilic food systems. Lipid-biomacromolecular compounds can be excellent carriers for carotenoid delivery by taking full advantage of the solubilization of lipids to non-polar nutrients and the water dispersion and gastrointestinal controlled release properties of biomacromolecules. This paper reviewed the research progress of lipid-biomacromolecular compounds as encapsulation and delivery carriers of carotenoids and summarized the material selection and preparation methods for biomacromolecular compounds. By considering the interaction between the two, this paper briefly discussed the effect of these compounds on carotenoid water solubility, stability, and bioavailability, emphasizing their delivery effect on carotenoids. Finally, various challenges and future trends of lipid-biomacromolecular compounds as carotenoid delivery carriers were discussed, providing new insight into efficient loading and delivery of carotenoids. [ABSTRACT FROM AUTHOR]

Previously, single classification models were mainly studied to classify human protein cell images, i.e., to identify a certain protein based on a set of different cells. However, a classifier can identify only one protein, in fact, a single cell usually consists of multiple proteins, and the proteins are not completely independent of each other. In this paper, we build a human protein cell classification model by multi-label learning. The logical relationship and distribution characteristics among the labels are analyzed to determine the different proteins contained in a set of different cells (i.e., containing multiple elements in the output space). In this paper, using human protein image data, we conducted comparison experiments on pre-trained Xception and InceptionResnet V2 to optimize the two models in terms of data augmentation, channel settings, and model structure. The results show that the Optimized InceptionResnet V2 model achieves high performance in the classification task. The final accuracy of the Optimized InceptionResnet V2 model we obtained reached 96.1%, which is a 2.82% improvement relative to that before the optimized model. [ABSTRACT FROM AUTHOR]

Integration and storage of bioactive reagents is an important and challenging task in microfluidic paper-based analytical devices (μPADs). Here, we developed a convenient and universal method to store proteins and preserve their activities in μPADs by using aqueous two-phase systems (ATPs) evolved film. A polyethylene glycol (PEG)–dextran (DEX) double-layer film was formed through dehydration of ATPs. Functional biomolecules were stored in the bottom DEX layer on the basis of the biased partitioning and rehydrated conveniently by simple addition of buffer solution at usage. As a demonstration, enzyme immunoassay (EIA) of carcinoembryonic antigen was performed successfully on μPAD integrated with antibodies. Even after 104 days of storage at 4 °C and ambient conditions, the EIA signal just lost less than 10% and 30%, which meet the storage requirements of invitro diagnosis reagents. The ATPs evolved double-layer film has double functions of stabilization and insulation, and provide a high efficiency of biomolecule preservation, thereby promoting the applications of μPADs in POC diagnostic assay. We present a universal and high efficient biomolecules preservation technique using the PEG-DEX ATPs evolved double-layer film for paper based microdevice. [Display omitted] • Antibodies and enzyme were integrated on the paper based microdevice using the ATPs evolved double-layer film. • Signals of μPADs with integrated immuno-reagents were maintained more than 90% after 104 days of storage at 4 °C conditions. • This protein storing method possessed simple operation, high compatibility, and high efficiency in preserving biomolecules. [ABSTRACT FROM AUTHOR]

Abstract: Dried blood spot samples (DBSS) on filter paper offer several advantages compared to conventional serum/plasma samples: they do not require any phlebotomy or separation of blood by centrifugation; they are less invasive; they allow sample stability and shipment at room temperature; and they pose a negligible risk of infection with blood-borne viruses, such as HIV, HBV and HCV, to those who handle them. Therefore dried blood spot samples (DBSS) on filter paper can be a quick, convenient and inexpensive means of obtaining blood samples for biomarker discovery, disease screening, diagnosis and treatment monitoring in non-hospitalized, public health settings. In this study, we investigated for the first time the potential application of dried blood spot samples (DBSS) in protein expression profiling using antibody array technology. First, optimal conditions for array assay performance using dried blood spot samples (DBSS) was established, including sample elution buffer, elution time, elution temperature and assay blocking buffer. Second, we analyzed dried blood spot samples (DBSS) using three distinct antibody array platforms, including sandwich-based antibody arrays, quantitative antibody arrays and biotin-label-based antibody arrays. In comparison with paired serum samples, detection of circulating proteins in dried blood spot samples (DBSS) correlated well for both low- and high-abundance proteins on all three antibody array platforms. In conclusion, our study strongly indicates the novel application of multiplex antibody array platforms to analyze dried blood spot samples (DBSS) on filter paper represents a viable, cost-effective method for protein profiling, biomarker discovery and disease screening in a large, population-based survey. [Copyright &y& Elsevier]

This paper proposes to teach students biology through dance. Knowledge of nucleotides, amino acids, and structures of DNA and protein is used to create dance music and choreography. The three nucleotides (A, T, G, or C) making up the amino acids of selected proteins are converted into a musical note. In accordance with DNA's double helix structure and base-pairing, the authors designed a low-cost, in-house Dance Dance Revolution (DDR) machine to allow for a dual player dance mode that requires students to synchronize their choreography during gameplay. [ABSTRACT FROM AUTHOR]

Simple Summary: The role of neurogenic locus notch homolog protein 1 (NOTCH1)- and nuclear factor erythroid 2-related factor 2 (NRF2)-related pathways is highly heterogeneous in small cell lung cancer (SCLC) and, when they are both abnormally activated, they can synergistically cause neoplastic proliferation. In this paper, we investigate the possible role of KEAP1 silencing in NRF2 and NOTCH axis deregulation in SCLC by evaluating its impact on the modulation of cellular defense systems, cell growth, and differentiation. We also investigate the impact of these systems' impairment in response to conventional chemotherapies and NOTCH inhibitors in silenced SCLC cell lines with different Kelch-like ECH-associated protein 1 (KEAP1) alterations. The KEAP1/NRF2 pathway is a master regulator of several redox-sensitive genes implicated in the resistance of tumor cells against therapeutic drugs. The dysfunction of the KEAP1/NRF2 system has been correlated with neoplastic patients' outcomes and responses to conventional therapies. In lung tumors, the growth and the progression of cancer cells may also involve the intersection between the molecular NRF2/KEAP1 axis and other pathways, including NOTCH, with implications for antioxidant protection, survival of cancer cells, and drug resistance to therapies. At present, the data concerning the mechanism of aberrant NRF2/NOTCH crosstalk as well as its genetic and epigenetic basis in SCLC are incomplete. To better clarify this point and elucidate the contribution of NRF2/NOTCH crosstalk deregulation in tumorigenesis of SCLC, we investigated genetic and epigenetic dysfunctions of the KEAP1 gene in a subset of SCLC cell lines. Moreover, we assessed its impact on SCLC cells' response to conventional chemotherapies (etoposide, cisplatin, and their combination) and NOTCH inhibitor treatments using DAPT, a γ-secretase inhibitor (GSI). We demonstrated that the KEAP1/NRF2 axis is epigenetically controlled in SCLC cell lines and that silencing of KEAP1 by siRNA induced the upregulation of NRF2 with a consequent increase in SCLC cells' chemoresistance under cisplatin and etoposide treatment. Moreover, KEAP1 modulation also interfered with NOTCH1, HES1, and DLL3 transcription. Our preliminary data provide new insights about the downstream effects of KEAP1 dysfunction on NRF2 and NOTCH deregulation in this type of tumor and corroborate the hypothesis of a cooperation of these two pathways in the tumorigenesis of SCLC. [ABSTRACT FROM AUTHOR]

Lysyl oxidase-like 2 (LOXL2) is a Cu2+-dependent amine oxidase, and it catalyzes the oxidative deamination of the ε-amino group of Lys residues on elastin and collagen, which is critical for extracellular matrix modeling. Stability is an important property for protein and it directly affects the dynamics and function of protein, which strongly relates to the human health and disease. However, the stability of LOXL2 is not well defined, i.e., which residues are important for its stability is unclear. In this paper, protein stability of LOXL2 was calculated by four methods, viz. mCSM, PremPS, DeepDDG and FoldX. Our results indicate that 95 residues (key residues) are important for the stability of LOXL2, with the amount of 14, 13, 8, 11, 26 in domain 1, 2, 3, 4, 5, respectively, while 23 in the linker between domains. Among these key residues, the types (numbers) are Leu (24), Val (20), Ile (12), Trp (11), Phe (11), Tyr (6), Cys (6), Met (4) and Arg (1). Most of the key residues are hydrophobic (88 in 95), which is consistent with the fact that hydrophobic interaction is the main force in protein folding. Cys residues are paired to constitute disulfide bonds, which play an important role in the structure and function of LOXL2. In the discussion, we compare the results with experimental data, and use the free energy perturbation for verification, which prove the reliability of this work. Thus, this study provides important insight for the research of disease treatment targeting LOXL2. [ABSTRACT FROM AUTHOR]

Simple Summary: The silkworm has fascinated people all around the world for centuries due to numerous applications of by-products resulting from its metamorphosis. A decline has been noticed due to industrialization and some disease concerns; however, several reasons strengthen compelling arguments for keeping up this species. The most prevalent nutrients identified along silkworm rearing, from mulberry leaves up to larvae, pupae, and silk, are proteins. The biological value of protein is based on amino acids. The presence of other nutrients, especially fatty acids (n-3 family), vitamins, minerals, and polyphenols, along with the profile in amino acids, which are taken from mulberry leaves and metabolized by larvae and then transferred to pupae, strengthen the conviction according to which silkworm by-products have a great nutritional potential that is either comparable with fish meal or better than other aquaculture-specific feedstuffs. This paper aims to present deep knowledge about amino acids (classification, metabolism, occurrence, mechanisms, and potential functions) for silkworms and fish to further understand these species. The silkworm Bombyx mori, the second most varied group of insects, is a fascinating insect that belongs to the Lepidoptera species. We aimed to deepen our knowledge about the composition and significance of amino acids (AA) from the sericulture chain to fish. AAs are the most prevalent molecules throughout the growth process of silkworms. We described AAs classification, occurrence, metabolism, and functions. Online datasets revealed that the essential AAs (EAA) level in fish meal and silkworm pupae (SWP) is comparable. SWP have a high content of methionine and lysine, which are the principal limiting AAs in fish diets, indicating that SWP have nutritional potential to be added to fish diets. Additionally, an overview of the data analyzed displays that SWP have a higher protein efficiency ratio than fish meal, the classical protein-rich source (>1.19 times), and compared to soybean meal, the second-most preferred source of protein in aquaculture (>2.08 times), indicating that SWP can be considered effective for animal feeding. In this study, we provide an overview of the current knowledge concerning AAs, paying special emphasis to EAAs and explaining, to some extent, certain mechanisms and functions of these compounds, from mulberry leaves to larvae–pupae and fish diets. [ABSTRACT FROM AUTHOR]

Objective: The purpose of this paper is to review the literature and compile promising and clinically relevant biomarkers in otolaryngology—head & neck surgery not related to autoimmune disorders. Study Design: Narrative review. Methods: PubMed and Google Scholar were queried using combined key words such as "biomarkers" and "otolaryngology." Additional queries were made with combined key words such as "biomarkers" and a particular subspecialty such as "rhinology" or "otology" to maximize yield of relevant titles. Subsequently, specific biomarkers identified, such as "beta-2 transferrin," were used as key words. Relevant titles were reviewed and selected for abstract review. Applicable abstracts were then selected for review of the full text. Results: Biomarkers currently in clinical use within the field of otolaryngology were included in this review. The compiled biomarkers were then detailed individually regarding their molecular characteristics, function, and clinical significance. Conclusions: The number of biomarkers in use in otolaryngology is rapidly expanding representing a new diagnostic modality for our field. This review defines the key biomarkers that are currently or likely to be soon translated into clinical use within the field of otolaryngology. The majority of these biomarkers are in the form of proteins such as beta-2 transferrin, thyroglobulin, and P16. Given their growing impact on diagnosis, management and surveillance of otolaryngologic disorders periodic surveys are needed for education and to guide further advances and applications of otolaryngologic biomarkers. [ABSTRACT FROM AUTHOR]

This article discusses the role of Tula-Family Proteins in cell signaling and activation. The Tula-Family Proteins, also known as UBASH3A and UBASH3B, are a relatively novel vertebrate gene/protein family that have been the subject of research for the past 20 years. Both UBASH3A and UBASH3B possess protein tyrosine phosphatase (PTP) activity and suppress protein tyrosine kinase (PTK)-dependent cell signaling. However, they also have distinct differences in substrate specificity, optimal conditions for PTP activity, and tissue expression. UBASH3A has been linked to autoimmune diseases, while UBASH3B appears to be involved in Behcet's disease. The article also discusses the conservation of the UBASH3 family in vertebrates and the unique UBASH3 family found in teleosts. Several papers published in this special issue further explore the role of UBASH3A/B proteins in immunity, autoimmune conditions, platelets, and individual development. The review articles highlight the challenges in understanding this family despite two decades of research. [Extracted from the article]

Summary: Maize grain is deficient in lysine. While the opaque2 mutation increases grain lysine, o2 is a transcription factor that regulates a wide network of genes beyond zeins, which leads to pleiotropic and often negative effects. Additionally, the drastic reduction in 19 kDa and 22 kDa alpha‐zeins causes a floury kernel, unsuitable for agricultural use. Quality protein maize (QPM) overcame the undesirable kernel texture through the introgression of modifying alleles. However, QPM still lacks a functional o2 transcription factor, which has a penalty on non‐lysine amino acids due to the o2 mutation. CRISPR/cas9 gives researchers the ability to directly target genes of interest. In this paper, gene editing was used to specifically target the 19 kDa alpha zein gene family. This allows for proteome rebalancing to occur without an o2 mutation and without a total alpha‐zein knockout. The results showed that editing some, but not all, of the 19 kDa zeins resulted in up to 30% more lysine. An edited line displayed an increase of 30% over the wild type. While not quite the 55% lysine increase displayed by QPM, the line had little collateral impact on other amino acid levels compared to QPM. Additionally, the edited line containing a partially reduced 19 kDa showed an advantage in kernel texture that had a complete 19 kDa knockout. These results serve as proof of concept that editing the 19 kDa alpha‐zein family alone can enhance lysine while retaining vitreous endosperm and a functional O2 transcription factor. [ABSTRACT FROM AUTHOR]

Proteomics is a field dedicated to the analysis of proteins in cells, tissues, and organisms, aiming to gain insights into their structures, functions, and interactions. A crucial aspect within proteomics is protein family prediction, which involves identifying evolutionary relationships between proteins by examining similarities in their sequences or structures. This approach holds great potential for applications such as drug discovery and functional annotation of genomes. However, current methods for protein family prediction have certain limitations, including limited accuracy, high false positive rates, and challenges in handling large datasets. Some methods also rely on homologous sequences or protein structures, which introduce biases and restrict their applicability to specific protein families or structures. To overcome these limitations, researchers have turned to machine learning (ML) approaches that can identify connections between protein features and simplify complex high-dimensional datasets. This paper presents a comprehensive survey of articles that employ various ML techniques for predicting protein families. The primary objective is to explore and improve ML techniques specifically for protein family prediction, thus advancing future research in the field. Through qualitative and quantitative analyses of ML techniques, it is evident that multiple methods utilizing a range of classifiers have been applied for protein family prediction. However, there has been limited focus on developing novel classifiers for protein family classification, highlighting the urgent need for improved approaches in this area. By addressing these challenges, this research aims to enhance the accuracy and effectiveness of protein family prediction, ultimately facilitating advancements in proteomics and its diverse applications. [ABSTRACT FROM AUTHOR]

Many results have been obtained when studying scientific papers citations databases in a network perspective. Articles can be ranked according to their current in-degree and their future popularity or citation counts can even be predicted. The dynamical properties of such networks and the observation of the time evolution of their nodes started more recently. This work adopts an evolutionary perspective and proposes an original algorithm for the construction of genealogical trees of scientific papers on the basis of their citation count evolution in time. The fitness of a paper now amounts to its in-degree growing trend and a “dying” paper will suddenly see this trend declining in time. It will give birth and be taken over by some of its most prevalent citing “offspring”. Practically, this might be used to trace the successive published milestones of a research field. [ABSTRACT FROM AUTHOR]

Required metrological tools, such as higher order reference measurements procedures, pure substance and matrix certified reference materials, are established for small well defined molecules. Difficulties still remain in the provision of SI traceable standards in the area of larger biomolecules such as peptides/proteins. The provision of Primary Calibration Reference Services has been identified as a core technical competency for National Measurement Institutes (NMIs). A concept has been elaborated by the Focus Group I on peptide/protein purity for the strategic planning of ongoing Key Comparisons of the Protein Analysis Working Group (PAWG) within the Comité Consultatif pour la Quantité de Matière (CCQM). The assignment of the mass fraction content of high purity peptide/protein materials will be the subject of the CCQM-K115/P55.2 comparison series to directly support NMI services and certified reference materials currently being provided by NMIs. [ABSTRACT FROM AUTHOR]

We studied the aging behaviour of gelatin used to size paper. Thus far, research on the aging of paper has largely ignored the sizing agent. Degradation of the protein was characterised and the impact of paper components, such as cellulose, and aluminium potassium sulphate was evaluated. Whatman No. 1 filter papers sized with two types of gelatins (A and B) were prepared as model samples. Commercially sized modern papers (Arches) were also studied in order to compare laboratory samples with real artist papers. Both types of papers were artificially aged (80 °C, 50% relative humidity for 35 and 94 days). Historic papers were included in the study in order to compare artificially aged with naturally aged gelatin. The aqueous extracts from the papers were characterised by aqueous size-exclusion chromatography (SEC) using four PL-Aquagel-OH columns and UV photodiode array detection at 220, 254 and 280 nm. Results showed that gelatin undergoes hydrolysis upon aging, type A gelatin showing a faster degradation rate than type B. The result was an increase in the lower-molar-mass fractions, under 50 000 g mol−1, and especially in a characteristic fraction with a peak molecular mass (Mp) of 14 000 g mol−1. A significant decrease in the extraction yields of α-, β- and γ-chains occurred after aging. This was attributed to crosslinking, leading to the formation of less-soluble polypeptides with very high molar mass (>800 000 g mol−1). Less than 10% alum had no impact on the degradation rate; higher alum contents accelerated hydrolysis reactions. [Copyright &y& Elsevier]

Simple Summary: Insects possess elastic cuticles in movable body parts such as the wing hinges and the basement of the hind legs of jumping insects. These cuticles contain a rubber-like protein called resilin. Resilin is a member of a cuticular protein family with a specific binding domain to chitin, which is another component of the cuticle. Resilin, or resilin-like proteins, have been reported in many insects. In a malaria vector mosquito, a curious cuticular protein has been found. It has a chitin-binding domain that is highly homologous to resilin, but it lacks repetitive sequences that are common in resilin and are essential for its elastic properties. It was unknown whether this protein is conserved in mosquitoes, whether this protein is the mosquito resilin, or whether mosquitoes possess another canonical resilin. In this paper, we found that this protein (termed resilin-related) is conserved in mosquitoes and seems to be evolutionarily derived from the resilin. Resilin-related, however, has very different structural features from resilin. Therefore, it probably plays special roles distinct from those of resilin in mosquitoes. Another resilin-like protein was found to exist commonly in holometabolous insects. Although this protein is likewise not a canonical resilin, it might confer elastic properties to cuticles in mosquitoes. Resilin is an elastic protein that is vital to insects' vigorous movement. Canonical resilin proteins possess the R&R Consensus, a chitin-binding domain conserved in a family of cuticular proteins, and highly repetitive sequences conferring elastic properties. In the malaria vector mosquito, Anopheles gambiae, however, a cuticular protein has been found that has an R&R Consensus resembling that of resilin but lacks the repetitive sequences (here, we call it resilin-related or resilin-r). The relationship between resilin-r and resilin was unclear. It was also unknown whether resilin-r is conserved in mosquitoes. In this paper, phylogenetic and structural analyses were performed to reveal the relationship of resilin homologous proteins from holometabolous insects. Their chitin-binding abilities were also assessed. A resilin-r was found in each mosquito species, and these proteins constitute a clade with resilin from other insects based on the R&R Consensus sequences, indicating an evolutionary relationship between resilin-r and resilin. The resilin-r showed chitin-binding activity as same as resilin, but had distinct structural features from resilin, suggesting that it plays specialized roles in the mosquito cuticle. Another resilin-like protein was found to exist in each holometabolous insect that possesses resilin-like repetitive sequences but lacks the R&R Consensus. These results suggest that similar evolutionary events occurred to create resilin-r and resilin-like proteins. [ABSTRACT FROM AUTHOR]

Debate surrounding the adverse consequences of feeding human-edible feedstuffs to livestock can be addressed through calculation of the net protein contribution (NPC) of the production system. If the NPC is greater than 1.0 for the production system, then there are net benefits from the system for human populations with an ever-increasing requirement for protein and amino acids. The aim of this paper was to calculate the NPC for an Australian pork supply chain on the basis of the unique characteristics of Australian ingredients. While calculation of NPC is not complex, intimate knowledge of the source of the nutrients and their quality, and interpretation of their human-edible protein fractions is essential if an accurate estimate is to be achieved. The NPC for an Australian pork supply chain was calculated using (a) actual, published or estimated values for human-edible fractions of feedstuffs, (b) the percentage of protein available within raw materials considered to be human edible, (c) recommended amino acid scoring patterns for infants, adolescents and adults, (d) published, and calculated from standard reference nutrient databases, digestible indispensable amino acid scores, (e) carcase yields and carcase composition from published studies, and (f) actual feed formulations, feed volumes and production data from a large Australian pork supply chain. The NPC for the assessed Australian pork supply chain was 3.26. This means the supply chain generates more than three times the human-edible protein it consumes in the process. This NPC is higher than previously published values, largely because of the composition of Australian pig diets, but demonstrates the positive value that livestock production systems make to human food supply. Livestock systems are often targeted as net consumers of vital nutrients such as protein and amino acids and the diversion of these nutrients from human diets. If production systems focus on the utilisation of waste streams, co-products and human-inedible feedstuffs, then they can make a net contribution to human-edible protein supply. Debate surrounding the adverse consequences of feeding human-edible feedstuffs to livestock can be addressed through calculation of the net protein contribution (NPC) of the production system. This paper calculates the NPC from an Australian pork supply chain to be 3.26. This number is higher than previously published values, largely because of the composition of Australian pig diets, but demonstrates the positive value that livestock production systems make to human food supply. [ABSTRACT FROM AUTHOR]

Simple Summary: There are three subfamilies of human PDZ-LIM family proteins with a total of 10 protein molecules, and PDZ-LIM family proteins serve as a class of scaffolding proteins that assume the function of signal transduction. In this paper, we describe the signature structural domains and major regulatory signals of PDZ-LIM family proteins and provide an overview and discussion of their functions in various tumors and major diseases, aiming to provide directions for future disease (mainly tumor) prevention and drug development. PDZ-LIM family proteins (PDLIMs) are a kind of scaffolding proteins that contain PDZ and LIM interaction domains. As protein–protein interacting molecules, PDZ and LIM domains function as scaffolds to bind to a variety of proteins. The PDLIMs are composed of evolutionarily conserved proteins found throughout different species. They can participate in cell signal transduction by mediating the interaction of signal molecules. They are involved in many important physiological processes, such as cell differentiation, proliferation, migration, and the maintenance of cellular structural integrity. Studies have shown that dysregulation of the PDLIMs leads to tumor formation and development. In this paper, we review and integrate the current knowledge on PDLIMs. The structure and function of the PDZ and LIM structural domains and the role of the PDLIMs in tumor development are described. [ABSTRACT FROM AUTHOR]

• Furan is used as a pro-reactive chemical handle to covalently capture an ODN target. • The electrophilic moiety that is generated in situ can react with naturally occurring targets. • DNA-DNA, DNA-RNA or DNA-Peptide crosslinking can be achieved with careful design of the probes. • Choice of the building block for the specific application is discussed. • Criteria for the optimal (photo)-oxidative activation of the furan are discussed. Oligonucleotides (ODNs) find applications as diagnostic and therapeutic tools due to their unique ability to interact, thanks to Watson-Crick base pairing, with a specific DNA or RNA target strand. Although most of the tools available today rely on mere hydrogen bond formation, chemical modifications to enable covalent interstrand-crosslinking (ICL) have been reported, and are gaining a place under the spotlight as they potentially offer a series of advantages over the state of the art, including a higher potency and selectivity. This methodological paper focuses on the use of a pro-reactive furan moiety and its subsequent oxidation for applications in ODN targeting. The design of effective capture and targeting probes to ensure high ICL yields is discussed and the mechanisms underlying the (photo)chemical oxidation of furan are explained. Furthermore, examples of furan-containing DNAs designed for different applications, including DNA-DNA or DNA-RNA ICL and DNA-peptide/protein targeting, are provided. The paper highlights the advantages of using different oxidative chemical triggers, such as N-bromosuccinimide or singlet oxygen, to offer additional selectivity control over the ICL reaction. [ABSTRACT FROM AUTHOR]

Paper has been proposed as an inexpensive and versatile carrier for microfluidics devices with abilities well beyond simple capillary action for pregnancy tests and the like. Unlike standard microfluidics devices, extracting a fluid from the paper is a challenge and a drawback to its broader use. Here, we extract fluid from narrow paper strips using surface acoustic wave (SAW) irradiation that subsequently atomizes the extracted fluid into a monodisperse aerosol for use in mass spectroscopy, medical diagnostics, and drug delivery applications. Two protein molecules, ovalbumin and bovine serum albumin (BSA), have been preserved in paper and then extracted using atomized mist through SAW excitation; protein electrophoresis shows there is less than 1% degradation of either protein molecule in this process. Finally, a solution of live yeast cells was infused into paper, which was subsequently dried for preservation then remoistened to extract the cells viaSAW atomization, yielding live cells at the completion of the process. The successful preservation and extraction of fluids, proteins and yeast cells significantly expands the usefulness of paper in microfluidics. [ABSTRACT FROM AUTHOR]

We present an origami paper-based electrophoretic device (oPAD-Ep) that achieves rapid (∼5 min) separation of fluorescent molecules and proteins. Due to the innovative design, the required driving voltage is just ∼10 V, which is more than 10 times lower than that used for conventional electrophoresis. The oPAD-Ep uses multiple, thin (180 μ/layer) folded paper layers as the supporting medium for electrophoresis. This approach significantly shortens the distance between the anode and cathode, and this, in turn, accounts for the high electric field (>1 kV/m) that can be achieved even with a low applied voltage. The multilayer design of the oPAD-Ep enables convenient sample introduction by use of a slip layer as well as easy product analysis and reclamation after electrophoresis by unfolding the origami paper and cutting out desired layers. We demonstrate the use of oPAD-Ep for simple separation of proteins in bovine serum, which illustrates its potential applications for point-of-care diagnostic testing. [ABSTRACT FROM AUTHOR]

A simple method is described for the patterned deposition of Teflon on paper to create an integrated platform for parallel organic synthesis and cell-based assays. Solvent-repelling barriers made of Teflon-impregnated paper confine organic solvents to specific zones of the patterned array and allow for 96 parallel flow-through syntheses on paper. The confinement and flow-through mixing significantly improves the peptide yield and simplifies the automation of this synthesis. The synthesis of 100 peptides ranging from 7 to 14 amino acids in length gave over 60 % purity for the majority of the peptides (>95 % yield per coupling/deprotection cycle). The resulting peptide arrays were used in cell-based screening to identify 14 potent bioactive peptides that support the adhesion or proliferation of breast cancer cells in a 3D environment. In the future, this technology could be used for the screening of more complex phenotypic responses, such as cell migration or differentiation. [ABSTRACT FROM AUTHOR]

• A paper-based multi-biocatalyst platform was successfully constructed. • Nanoporous gold with unique properties was used as supporting material. • Three different markers can be integrally detected by the multi-biocatalyst platform. • The reliable detection of L. monocytogenes was successfully realized. As biomarkers for pathogenic Listeria monocytogenes , the detection of gene hly , acetoin, and listeriolysin O (LLO) is of great significance for preventing L. monocytogenes infection and diagnosing Listeriosis. Here, a portable paper-based multi-biocatalyst platform was constructed to identify L. monocytogenes by detecting multiple biomarkers at different levels: gene hly (nucleic acid), acetoin (small molecule metabolite), and LLO (protein). The integrating detections of the three biomarkers were successfully performed by two different modified working electrodes on a single paper-based multi-biocatalyst platform. The sensitive and reliable identification of L. monocytogenes was achieved using the portable paper-based multi-biocatalyst platform with wider detection range (from 1.0 × 104 to 1.0 × 109 CFU mL−1) and lower detection limit (104 CFU mL−1). Additionally, the simpler detection procedure and shorter detection time (2 h) endows the portable platform a better detection performance compared with National Food Safety Standard Method (China-GB 4789.30-2016). Moreover, the results detected by the portable platform in spiked samples were consistent with that obtained by the real-time quantitative polymerase chain reaction method, indicating a good application potential. These unique characteristics suggest that the portable platform provides a reliable identification method for L. monocytogenes. [ABSTRACT FROM AUTHOR]

Active cellulose-based packaging materials were prepared by binding lactoferrin to paper modified with anionic polyelectrolytes. About 63% of the protein added in the slurry was present in the finished paper, corresponding to about 0.28 mg protein per cm2 of the glassine paper produced. The efficacy of the activated materials was assessed by microbiological trials, employing targets bacteria considered as spoilage and/or pathogens in food products. Lactoferrin-activated paper evidenced its efficacy against growing cells of E. coli and L. innocua, with a maximum of 0.3 and 1.3 log cycles reduction of the final population, respectively. These results can be considered a promising starting point and will be the focus of further research, aimed at developing innovative cellulose-based active food packaging. [ABSTRACT FROM AUTHOR]

Given their simplicity and functionality, paper-based microfluidic systems are considered to be ideal and promising bioassay platforms for use in less developed countries or in point-of-care services. Although a series of innovative techniques have recently been demonstrated for the fabrication of such platforms, development of simple, inexpensive and versatile new strategies are still needed in order to reach their full potential. In this communication, we describe a simple yet facile approach to fabricate paper-based sensor platforms with a desired design through a vapor-phase polymer deposition technique. We also show that the fabricated platforms could be readily employed for the detection of various biological target molecules including glucose, protein, ALP, ALT, and uric acid. The limit of detection for each target molecule was calculated to be 25 mg dL−1 for glucose, 1.04 g L−1 for protein, 7.81 unit per L for ALP, 1.6 nmol L−1 for ALT, and 0.13 mmol L−1 for uric acid. [ABSTRACT FROM AUTHOR]

In this work, we developed a microfluidic paper-based analytical device (μPAD) for the on-line isolation of proteins and the electrochemical detection of lead ions (Pb(II)) in urine samples. The patterned filter paper was prepared through the direct printing of microchannel patterns on filter paper using an office laser printer. The paper was modified with protein precipitant and was then coupled with a detachable three-electrode system. Experimental parameters, namely, modification reagents, microchannel length and width, deposition potential, and deposition time, were optimized. Then, the maximum protein concentration under which the device can function was obtained as 300 mg L−1. The linear range was 10–500 μg L−1 with a detection limit of 9 μg L−1. The effectiveness of this device was demonstrated through the quantification of Pb(II) in urine samples and the results agreed with those of atomic absorption spectrometry (AAS). • A simple electrochemical paper-based microfluidic device was built using laser printer and 3D printer. • The device enables on-line isolation of proteins in urine. • The maximum protein concentration under which the proposed device can function was 300 mg L−1. • Direct electrochemical detection of Pb in urine was achieved with a detection limit of 9 μg L−1. [ABSTRACT FROM AUTHOR]

Abstract: This paper described a convenient semiquantitative method for colorimetric detection of protein with self-calibration integrated on the test strip. Hydrophilic paper was employed as microfluidic device for running colorimetric assay, tree-shaped design was developed to ensure uniform microfluidic flow for multiple branches. The approach was validated with bovine serum albumin (BSA) colorimetric detection, and colorimetric results observed by naked eyes were consistent with that from apparatus. The device could be coupled with digital transmission of images for remote monitoring system for diagnosis, food control, and environmental analysis. [Copyright &y& Elsevier]

Here, we report a strategy for the design of an inexpensive paper analytical device (PAD) for quantitative detection of oligonucleotides and proteins. Detection is based on the principle of target-induced conformational switching of an aptamer linked to an electrochemical label. This simple and robust method is well matched to the equally simple and robust characteristics of the PAD platform. The demonstrated limits of detection for DNA and thrombin are 30 nM and 16 nM, respectively, and the device-to-device reproducibility is better than ±10%. The PAD has a shelf life of at least 4 weeks, involves little user intervention, and requires a sample volume of just 20 μl. [ABSTRACT FROM AUTHOR]

Paper spray (PS) ionization, an ambient ionization method, has previously been explored as a direct and fast method for mass spectrometric analysis of complex mixtures. It has been applied to the analysis of a wide variety of compounds, mostly small molecules. The work reported here extends the application of PS ionization to noncovalent protein complexes on an ion mobility tandem mass spectrometer. Similar mass spectra for protein complexes were obtained by PS ionization and nanoflow electrospray ionization (nESI), indicating that intact protein complexes can be preserved in PS ionization. In addition, collisional cross sections measured by ion mobility provide evidence that the protein assemblies may remain compact by PS ionization. With PS, it is possible to detect hemoglobin tetramer from a blood sample with minimal sample preparation. This is the first report to show that PS ionization is a promising ionization method for nonconvalent protein complexes. [ABSTRACT FROM AUTHOR]

NanoFAST is the smallest fluorogen-activating protein, consisting of only 98 amino acids, used as a genetically encoded fluorescent tag. Previously, only a single fluorogen with an orange color was revealed for this protein. In the present paper, using rational mutagenesis and in vitro screening of fluorogens libraries, we expanded the color palette of this tag. We discovered that E46Q is one of the key substitutions enabling the range of possible fluorogens to be expanded. The introduction of this and several other substitutions has made it possible to use not only orange but also red and green fluorogens with the modified protein. [ABSTRACT FROM AUTHOR]

Leishmania tarentolae (LEXSY) system is an inexpensive and effective expression approach for various research and medical purposes. The stated advantages of this system are the possibility of obtaining the soluble product in the cytoplasm, a high probability of correct protein folding with a full range of post-translational modifications (including uniform glycosylation), and the possibility of expressing multi-subunit proteins. In this paper, a LEXSY expression system has been employed for obtaining the receptor binding domain (RBD) of the spike-protein of the SARS-CoV-2 virus and the homopentameric acetylcholine-binding protein (AChBP) from Lymnaea stagnalis. RBD is actively used to obtain antibodies against the virus and in various scientific studies on the molecular mechanisms of the interaction of the virus with host cell targets. AChBP represents an excellent structural model of the ligand-binding extracellular domain of all subtypes of nicotinic acetylcholine receptors (nAChRs). Both products were obtained in a soluble glycosylated form, and their structural and functional characteristics were compared with those previously described. [ABSTRACT FROM AUTHOR]

β-glucosidase is an essential enzyme for the enzymatic hydrolysis of lignocellulosic biomass, as it catalyzes the final stage of cellulose breakdown, releasing glucose. This paper aims to produce β-glucosidase from Saccharomyces cerevisiae and evaluate the enzymatic degradation of delignified sugarcane bagasse. S. cerevisiae was grown in yeast peptone dextrose medium. Partial purification of the enzyme was achieved through precipitating proteins with ethanol, and the optimal activity was measured by optimizing pH and temperature. The effects of ions, glucose tolerance, and heat treatment were evaluated. Delignified sugarcane bagasse was hydrolyzed by the enzyme. β-glucosidase showed a specific activity of 14.0712 ± 0.0207 U mg−1. Partial purification showed 1.22-fold purification. The optimum pH and temperature were 6.24 and 54 °C, respectively. β-glucosidase showed tolerance to glucose, with a relative activity of 71.27 ± 0.16%. Thermostability showed a relative activity of 58.84 ± 0.91% at 90 °C. The hydrolysis of delignified sugarcane bagasse showed a conversion rate of 87.97 ± 0.10% in the presence of Zn2+, an ion that promoted the highest increase in enzymatic activity. S. cerevisiae produced an extracellular β-glucosidase with good stability at pH and temperatures conventionally applied in the hydrolysis of lignocellulosic biomass, showing viability for industrial application. [ABSTRACT FROM AUTHOR]

Crystallization is an important unit operation in industry, and proteins are widely studied owing to their biological activity and multiple functions. Protein crystals can not only facilitate the elucidation of protein structural information but also have important prospects in biopharmaceuticals. However, owing to their high molecular weight and complex structure, most proteins are difficult to crystallize or the crystallization processes are difficult to control effectively. This paper aims to provide an overview of recent reinforcement techniques for protein crystallization. Several important additive applications and principles in protein crystallization are comprehensively reviewed, including ionic liquids (ILs), deep eutectic solvents (DESs), and molecularly imprinted polymers (MIPs). The mechanism of external fields, including magnetic, electric, ultrasonic, shear, and light fields, enhancing protein crystallization are described and summarized. Finally, the challenges and future research prospects of protein crystallization are proposed. This work is expected to guide the rational design of the protein crystallization process. [ABSTRACT FROM AUTHOR]