Protein Activity Research Articles

Activity and phosphatidylcholine transfer protein interactions of skeletal muscle thioesterase Them2 enable hepatic steatosis and insulin resistance

Thioesterase superfamily member 2 (Them2), a long-chain fatty acyl-CoA thioesterase that is highly expressed in oxidative tissues, interacts with phosphatidylcholine transfer protein (PC-TP) to regulate hepatic lipid and glucose metabolism and to suppress insulin signaling. High-fat diet (HFD)-fed mice lacking Them2 globally or specifically in skeletal muscle, but not liver, exhibit reduced hepatic steatosis and insulin resistance. Here, we report that the capacity of Them2 in skeletal muscle to promote hepatic steatosis and insulin resistance depends on both its catalytic activity and interaction with PC-TP. Two residues of Them2 catalytic site were mutated (N50A/D65A) to produce the inactive enzyme while maintaining its homotetrameric structure and interaction with PC-TP. Restoration of skeletal muscle expression in Them2-/- mice using recombinant adeno-associated virus revealed that wild-type (WT), but not N50A/D65A Them2, promoted HFD-induced weight gain and hepatic steatosis. This was accompanied by greater impairment of insulin sensitivity in WT compared with N50A/D65A Them2. Pharmacological inhibition or genetic ablation of PC-TP attenuated these effects. In reductionist experiments, conditioned medium collected from WT primary cultured myotubes promoted excess lipid accumulation in oleic acid-treated primary cultured hepatocytes relative to Them2-/- myotubes, which was attributable to secreted extracellular vesicles (EV). Reconstitution of Them2 expression in Them2-/- myotubes affirmed the requirements for catalytic activity and PC-TP interactions for EV to promote lipid accumulation in hepatocytes. These studies provide valuable mechanistic insights whereby Them2 in skeletal muscle promotes hepatic steatosis and establish both Them2 and PC-TP as represent attractive targets for managing metabolic dysfunction-associated steatotic liver disease.

Impact of Feeding Artemia franciscana Enriched with Various Oil Resources on Growth, Blood Biochemical and Behavioral Indices, and Survival of Oreochromis Niloticus

Abstract The main objective of the present trial was to examine the efficacy of feeding tilapia fry fish on Artemia franciscana diets supplemented with various oil emulsion resources in terms of performance, behavior indices, survival rate, blood biochemical parameters, and immunological response. Four hundred Nile tilapia fry (weighing 0.15±0.05 g and measuring 2.17±0.08 cm) were randomly allocated into four equal groups (each with five repetitions) and acclimatized for fifteen days. The first group served as the control and received unenriched Artemia franciscana (G0), while the remaining three groups were fed Artemia franciscana diets enriched with different oil resources (0.5 mL oil per liter for 6 hours): soybean oil (G1), sesame oil (G2), and rice bran oil (G3). Behavioral observations were recorded during the 45-day experimental period. At the end of the feeding trial, the chemical composition and fatty acid content of both Artemia and fingerlings were analyzed. Furthermore, the growth performance, survival, and immune response of the fingerlings were evaluated. The results indicated noticeable improvements in behavioral measurements (feeding, foraging and schooling), performance (final length, final weight, net weight gain, feed conversion ratio and specific growth rate), survival, and immune response among fry fish supplemented with enriched Artemia, particularly those enriched with soybean oil. Additionally, the chemical composition and fatty acid content of both Artemia and fish fry were significantly enhanced when oil emulsions are applied, with soybean oil demonstrating the most prominent improvements. Whereas, supplementing fry fish Artemia diets with oil resulted in lower liver enzyme activity and higher protein component levels in plasma in comparison to the control group. In brief, feeding Nile tilapia fry fish Artemia diets enriched with a soybean oil emulsion (0.5 mL/L) is recommended for promoting high performance, immunological activity, and survival throughout the early stage till fingerlings phase.

Roles of cytochromes P450 and ribosome inhibition in the interaction between two preoccupying mycotoxins, aflatoxin B1 and deoxynivalenol

Mycotoxins are a threat to human and animal health. Climate change increases their occurrence and our dietary exposure. Although humans and animals are concomitantly exposed to several mycotoxins, their combined effects are poorly characterised. This study investigated the interaction between aflatoxin B1 (AFB1), the most potent natural carcinogen, and deoxynivalenol (DON), which is among the most prevalent mycotoxins. AFB1 is associated with hepatocellular carcinoma through its bioactivation by cytochrome P450 (CYP450) enzymes; while DON induces ribotoxic stress leading to an alteration of intestinal, immune and hepatic functions. Analysis of DNA damage biomarkers γ-H2AX and 53BP1 revealed that DON reduces the genotoxicity of AFB1. This effect was mimicked with cycloheximide, another ribosome inhibitor; moreover DOM-1, a DON-derivative lacking ribosome inhibition, did not affect DNA damage. Exposure to DON, alone or in combination with AFB1, decreased the protein levels and/or activities of CYP1A2 and CYP3A4 in a time- and dose-dependent manner. Altogether, these results revealed an original interaction between DON and AFB1, DON inhibiting the genotoxicity of AFB1. The underlying mechanism involves ribosome inhibition by DON and the subsequent impairment of CYP450s, responsible for the bioactivation of AFB1. This work highlights the importance of studying mycotoxins not only individually but also in mixture and of considering food contaminants as part of the exposome.

H2S protects rat cerebral ischemia-reperfusion injury by inhibiting expression and activation of hippocampal ROCK2 at the Thr436 and Ser575 sites

BackgroundH2S is an endogenous gas signal molecule, which protects cerebral ischemia/reperfusion (I/R) injury by phosphorylating rho-associated coiled coil-containing protein kinase 2 (ROCK2) at Tyr722, and inhibiting ROCK2 protein expression and activities. We previously reported that H2S protected rat neurons from hypoxia/reoxygenation injury in vitro through inhibiting phosphorylation of ROCK2 at Thr436 and Ser575, but it is unclear whether these two sites are involved in protection of H2S against cerebral I/R injury. MethodRats transfected with wild-type and mutant eukaryotic plasmids of ROCK2 in hippocampus were used to establish I/R model by ligating bilateral common carotid artery. Rat behavioral deficit was detected by water maze assay, and ROCK2, lactate dehydrogenase (LDH), nerve-specific enolase (NSE) and reactive oxygen species (ROS) were determined by ELISA. ROCK2 expressions was examined by western-blot assay, and bcl-2 and Bax mRNAs were examined by RT-qPCR. ResultsNaHS (4.8mg/kg) significantly inhibited the I/R-increased serum LDH, NSE and ROS in the ROCK2wild-pEGFP-N1-transfected rats, but had no obvious effect in the ROCK2T436A-pEGFP-N1- or the ROCK2S575F-pEGFP-N1-transfected rats; inhibitions of NaHS on the I/R-increased escape latency and the I/R-decreased percentage of target quadrant distance to total distance were markedly attenuated or abolished in the ROCK2T436A-pEGFP-N1- or the ROCK2S575F-pEGFP-N1-transfected rats compared with those in the ROCK2wild-pEGFP-N1-transfected rats; NaHS obviously inhibited the I/R-increased hippocampal ROCK2 and GFP-ROCK2 proteins, Bax mRNA, and ROCK2 activity, as well as the I/R-decreased hippocampal bcl-2 mRNA in the hippocampus of the ROCK2wild-pEGFP-N1-transfected rats, but had no significant effect in the ROCK2T436A-pEGFP-N1- or the ROCK2S575F-pEGFP-N1-transfected rats. ConclusionH2S protects cerebral I/R injury in rats by inhibiting expression and activation of hippocampal ROCK2 via the Thr436 and Ser575 sites.

Green synthesis of Au nanoparticles by Scutellaria barbata extract for chemo-photothermal anticancer therapy

IntroductionThe combination of photothermal therapy and chemotherapy has emerged as a promising strategy for cancer treatment. The green synthesis of gold nanoparticles (Au NPs) using extracts from traditional Chinese medicine Scutellaria barbata D. Don (Scu) has revealed an economical, sustainable, and eco-friendly approach for the outstanding anticancer activities. MethodsScu extract was mixed with HAuCl4 solution to fabricate Scu-Au NPs. The synthesis process was optimized by varying the reaction temperature, reaction time, and HAuCl4 concentration. The changes in the components of Scu before and after synthesis were analyzed using the 3,5-dinitrosalicylic acid (DNS) reduction test, the rutin colorimetric method, and the Folin–Ciocalteu method. The morphology, size, and structure of Scu-Au NPs were characterized by TEM, DLS, XRD, and XPS. The photothermal characteristics induced by near-infrared (NIR) irradiation were assessed by continuously monitoring the temperature elevation. The anticancer activity and mechanisms of Scu-Au NPs were investigated using MTT assay, cell uptake studies, ROS level detection, cell apoptosis and necrosis assays, flow cytometry analysis, intracellular Caspase-3 protein activity fluorescence detection, and Western blot analysis on A549, 4T1, and RAW264.7 cells. ResultsScu-Au NPs were successfully fabricated using Scu extract as a reducing agent. The Scu-Au NPs were spherical with a size of 50 nm. During the synthesis process, the levels of reducing sugars, flavonoids, and polyphenols in the Scu extract decreased by 80.8%, 54.3%, and 70.1%, respectively. Scu-Au NPs demonstrated excellent photothermal responsiveness, thermostability, and biocompatibility. For chemo-photothermal anticancer therapy, the cytotoxicity of Scu-Au NPs on 4T1 cells reached approximately 85% upon exposure to 808 nm NIR irradiation. The anticancer activity was attributed to the induction of apoptosis and necrosis, achieved by increasing intracellular ROS levels, causing cell cycle arrest at the S phase, and modulating the expression of apoptosis-related proteins. DiscussionOur synthesized Scu-Au NPs exhibit favorable photothermal conversion characteristics, demonstrating excellent therapeutic efficacy and safety for cancer treatment.

Integrated transcriptomic and proteomic analysis revealed the regulatory role of 5-azacytidine in kenaf salt stress alleviation

Salinity stress limits agricultural production. The DNA methyltransferase inhibitor, 5-azacitidine (5-azaC), plays a role in plant abiotic stress regulation, but its molecular basis in mediating salinity tolerance in kenaf remains unclear. To investigate the effects on 5-azaC on alleviating salt stress, kenaf seedlings were pre-treated with 0, 50, 100, 150, and 200 μM 5-azaC and then exposed to 150 mM NaCl in a nutrient solution. Physiological, transcriptomic, and proteomic analyses were conducted on the root system to understand the regulatory mechanism of 5-azaC (comparing 5-azaC150 and control group 5-azaC0) under salt stress. The results indicated that 5-azaC significantly mitigated salt stress in kenaf by activating the antioxidant system, reducing reactive oxygen species (ROS), and increasing starch, soluble sugars, and adenosine triphosphate (ATP) content. A total of 14,348 differentially expressed genes (DEGs) and 313 differentially abundant proteins (DAPs) were identified. Combined proteomic and transcriptomic analysis revealed 27 DEGs/DAPs, with jointly up-regulated proteins (genes) including HcTHI1, HcBGLU11, and HcCBL1, and jointly down-regulated proteins (genes) including HcGAPDH, HcSS, and HcPP2C52. Overexpression and virus-induced gene silencing (VIGS) of HcPP2C52 demonstrated its role as a negative regulator of salt tolerance. These findings provide insights into the regulatory role of 5-azaC in plant responses to abiotic stresses. SignificanceThe specific molecular mechanism by which 5-azaC affects gene expression and protein activity of kenaf has been revealed, leading to enhanced salt tolerance.

Perubahan Komponen Kimia Fungsional pada Umbi, Tepung, dan Beras Analog Ubi Jalar Ungu

The purple sweet potato tuber, rich in functional chemical components, offers potential for development into high-value products such as flour, which can then be processed into analog rice. This research aims to evaluate the changes in functional chemical components–namely moisture, ash, protein, fat, carbohydrates, anthocyanin, and β-carotene–along with antioxidant activity during the processing of purple sweet potato tubers into flour and analog rice. The chemical composition of the tubers was found to be as follows: moisture content of 78.00% on a wet basis (wb), ash content of 9.70% on a dry basis (db), fat content of 5.47% (db), protein content of 26.93% (db), carbohydrate content of 58.41% (db), anthocyanin content of 31.79 mg/100 g (db), β-carotene content of 17.73 mg/100 g (db), and antioxidant activity of 65.70%. The composition of the resulting flour included water (6.71% wb), ash (2.40% db), fat (1.06% db), protein (3.80% db), carbohydrate (92.75% db), anthocyanin (3.47 mg/100 g db), β-carotene (1.51 mg/100 g db), and antioxidant activity (45.59%). The chemical content of the analog rice was as follows: water (8.43% wb), ash (1.78% db), fat (1.62% db), protein (4.13% db), carbohydrate (92.50% db), anthocyanin (0.71 mg/100 g db), β-carotene (1.05 mg/100 g db), and antioxidant activity (8.38%). Processing the tubers into flour resulted in a reduction of fat, protein, anthocyanins, β-carotene, and antioxidant activity, while carbohydrate content increased. Similarly, the conversion of flour into analog rice led to decreased levels of carbohydrates, anthocyanins, β-carotene, and antioxidant activity, whereas fat and protein levels increased after transfor-mation into analog rice. The processing stages that significantly contributed to these changes in chemical components included drying, formulation, and extrusion.

Characterization of a β-carotene isomerase from the cyanobacterium Cyanobacteria aponinum.

Carotenoids are essential components of the photosynthetic apparatus and precursors of plant hormones, such as strigolactones (SLs). SLs are involved in various aspects of plant development and stress-response processes, including the establishment of root and shoot architecture. SL biosynthesis begins with the reversible isomerization of all-trans-carotene into 9-cis-β-carotene, catalysed by DWARF27 β-carotene isomerase (D27). Sequence comparisons have revealed the presence of D27-related proteins in photosynthetic eukaryotes and cyanobacteria lacking SLs. To gain insight into the evolution of SL biosynthesis, we characterized the activity of a cyanobacterial D27 protein (CaD27) from Cyanobacterim aponinum, using carotenoid-accumulating Escherichia coli cells and in vitro enzymatic assays. Our results demonstrate that CaD27 is an all-trans/cis and cis/cis-β-carotene isomerase, with a cis/cis conversion preference. CaD27 catalysed 13-cis/15-cis-, all-trans/9-cis-β-carotene, and neurosporene isomerization. Compared with plant enzymes, it exhibited a lower 9-cis-/all-trans-β-carotene conversion ratio. A comprehensive genome survey revealed the presence of D27 as a single-copy gene in the genomes of 20 out of 200 cyanobacteria species analysed. Phylogenetic and enzymatic analysis of CaD27 indicated that cyanobacterial D27 genes form a single orthologous group, which is considered an ancestral type of those found in photosynthetic eukaryotes. This article is part of the theme issue 'The evolution of plant meta‌bolism'.

The Role of Thioredoxin System in Shank3 Mouse Model of Autism.

Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder characterized by difficulties in social interaction and communication, repetitive behaviors, and restricted interests. Unfortunately, the underlying molecular mechanism behind ASD remains unknown. It has been reported that oxidative and nitrosative stress are strongly linked to ASD. We have recently found that nitric oxide (NO•) and its products play an important role in this disorder. One of the key proteins associated with NO• is thioredoxin (Trx). We hypothesize that the Trx system is altered in the Shank3 KO mouse model of autism, which may lead to a decreased activity of the nuclear factor erythroid 2-related factor 2 (Nrf2), resulting in oxidative stress, and thus, contributing to ASD-related phenotypes. To test this hypothesis, we conducted in vivo behavioral studies and used primary cortical neurons derived from the Shank3 KO mice and human SH-SY5Y cells with SHANK3 mutation. We showed significant changes in the levels and activity of Trx redox proteins in the Shank3 KO mice. A Trx1 inhibitor PX-12 decreased Trx1 and Nrf2 expression in wild-type mice, causing abnormal alterations in the levels of synaptic proteins and neurotransmission markers, and an elevation of nitrosative stress. Trx inhibition resulted in an ASD-like behavioral phenotype, similar to that of Shank3 KO mice. Taken together, our findings confirm the strong link between the Trx system and ASD pathology, including the increased oxidative/nitrosative stress, and synaptic and behavioral deficits. The results of this study may pave the way for identifying novel drug targets for ASD.

Asparagine as a signal for glutamine sufficiency via asparagine synthetase: a fresh evidence-based framework in physiology and oncology.

Among the 20 proteinogenic amino acids, glutamine (GLN) and asparagine (ASN) represent a unique cohort in containing a terminal amide in their side chain, and share a direct metabolic relationship, with glutamine generating asparagine through the ATP-dependent asparagine synthetase (ASNS) reaction. Circulating glutamine levels and metabolic flux through cells and tissues greatly exceed those for asparagine, and "glutamine addiction" in cancer has likewise received considerable attention. However, historic and recent evidence collectively suggest that in spite of its modest presence, asparagine plays an outsized regulatory role in cellular function. Here, we present a unifying evidence-based hypothesis that the amides constitute a regulatory signaling circuit, with glutamine as a driver and asparagine as a second messenger that allosterically regulates key biochemical and physiological functions, particularly cell growth and survival. Specifically, it is proposed that ASNS serves as a sensor of substrate sufficiency for S-phase entry and progression in proliferating cells. ASNS-generated asparagine serves as a subsequent second messenger that modulates the activity of key regulatory proteins and promotes survival in the face of cellular stress, and serves as a feed-forward driver of S-phase progression in cell growth. We propose that this signaling pathway be termed the amide signaling circuit (ASC) in homage to the SLC1A5-encoded ASCT2 that transports both glutamine and asparagine in a bidirectional manner, and has been implicated in the pathogenesis of a broad spectrum of human cancers. Support for the ASC model is provided by the recent discovery that glutamine is sensed in primary cilia via ASNS during metabolic stress.

Nutrient profile, protease and cellulase activities of protein extracted from black soldier fly (Hermetia illucens) larvae reared on various substrates

The Black Soldier Fly (BSF; Hermetia illucens) larvae are recognized for their ability to convert diverse organic materials into protein-rich biomass, depending on the substrate they consume. The composition of these substrates can significantly impact the nutrient profile and enzyme activities of the resulting maggot protein extract (MPE). Therefore, this exploratory research aimed to assess the nutrient content, protease, and cellulase activity of MPE obtained from BSF maggots reared on different substrates, with a specific focus on substrates A (comprising restaurant waste and rejected milk), B (layer manure), and C (kitchen waste). The results showed that maggot meal from layer manure had the highest protein content (45.36%) and the lowest fat content (18.44%). Amino acids in maggot meal contained high levels of glutamic acid, aspartic acid, alanine, valine, leucine, and isoleucine. Lauric acids were found in maggot meal from kitchen waste (33.79%), layer manure (32.18%), and restaurant waste and rejected milk (22.94%). Maggot meal from layer manure had the highest oleic acid content (15.13%). The protein concentration of MPE from various substrates ranged from 0.56 to 0.601 mg/ml (at 60% w/v ammonium sulfate saturation) and 0.555 to 0.609 mg/ml (at 70% ammonium sulfate saturation). The protease activity of MPE from layer manure substrates exhibited optimum activity and stability in neutral to alkaline pH, with activity levels of 0.748 U/mg at pH 7.0 and pH 11.0 (at 60% w/v ammonium sulfate saturation) and 0.774 units/mg at 70% w/v ammonium sulfate saturation. The highest cellulase activity was found in MPE from kitchen waste, which remained stable at pH 5.0-11.0. In general, maggots from different substrate sources exhibited distinct nutrient profiles and enzyme activities. Protein extract from maggots grown in layer manure showed the most suitable nutrient profile for use as an alternative source of protein feed and protease enzymes.

The Role of Protein Post-Translational Modifications in Fruit Ripening

Fruit ripening represents a multifaceted biological process intricately controlled by an array of plant hormones, transcription factors, and epigenetic modifications. These regulatory mechanisms are crucial in determining fruit quality and post-harvest shelf life. Recent advancements in proteomics have shifted the focus toward understanding protein post-translational modifications (PTMs), which play a crucial role in modulating protein function. PTMs enhance protein activity and stability by altering their properties after biosynthesis, thereby adding an additional layer of regulation to the ripening process. This paper provides a comprehensive review of the roles of PTMs, including ubiquitination, phosphorylation, redox modifications, and glycosylation in regulating fruit ripening. Emphasis is placed on the intricate interplay between these PTMs and key regulator factors such as plant hormones, transcriptional mechanisms, and epigenetic modifications. By exploring these interactions, this review seeks to enhance our understanding of the complex regulatory network underlying fruit ripening and to offer novel perspectives on strategies for fruit preservation.

Dietary inflammatory index, physical activity, body composition, and high sensitivity c-reactive protein levels in adolescent athletes in Central Java

Inflammation is the body's response to tissue damage and infection. Prolonged inflammation can increase the risk of non-communicable diseases, including cardiovascular disease. Some factors that can trigger inflammation are dietary patterns, high physical activity, and body composition. This study examined the correlation between dietary inflammatory index (DII), physical activity, body composition, and high sensitivity C-reactive protein (hs-CRP) levels. It examined differences in characteristics based on gender. This study was a cross-sectional design of 55 adolescent athletes at the Student Sports Education and Training Center in Central Java. The exclusion criteria were an athlete who had a physical injury (did not participate in training) and a smoker. The DII score was calculated based on the dietary pattern using SQFFQ, anthropometric data (weight, height, and nutritional status based on BMI/age), body composition (body fat percentage, fat mass, and fat-free mass) using the BIA tool, physical activity data for the last seven days using IPAQ and hs-CRP levels using the FIA method. Our result showed that the DII score was significantly correlated with hs-CRP (p=0.013). There was no correlation between physical activity, body composition, and anthropometry with hs-CRP levels (p0.05). There were differences in body height, body weight, body fat percentage, and fat-free mass between males and females (p0.001). The diet of athletes had anti-inflammatory potential (83.6%), as evidenced by the intake of vitamins and minerals, and athletes had low hs-CRP levels 1 mg/L (85.5%), indicating that no inflammation occurred. Future longitudinal research is needed with a large population of adolescent athletes to explain the role of anti-inflammatory and pro-inflammatory diets on the incidence of inflammation and measure physical exercise in athletes to reduce inflammatory bias.

Revolutionizing Molecular Design for Innovative Therapeutic Applications through Artificial Intelligence

The field of computational protein engineering has been transformed by recent advancements in machine learning, artificial intelligence, and molecular modeling, enabling the design of proteins with unprecedented precision and functionality. Computational methods now play a crucial role in enhancing the stability, activity, and specificity of proteins for diverse applications in biotechnology and medicine. Techniques such as deep learning, reinforcement learning, and transfer learning have dramatically improved protein structure prediction, optimization of binding affinities, and enzyme design. These innovations have streamlined the process of protein engineering by allowing the rapid generation of targeted libraries, reducing experimental sampling, and enabling the rational design of proteins with tailored properties. Furthermore, the integration of computational approaches with high-throughput experimental techniques has facilitated the development of multifunctional proteins and novel therapeutics. However, challenges remain in bridging the gap between computational predictions and experimental validation and in addressing ethical concerns related to AI-driven protein design. This review provides a comprehensive overview of the current state and future directions of computational methods in protein engineering, emphasizing their transformative potential in creating next-generation biologics and advancing synthetic biology.

Modeling Of the Structure of the tRolC Protein of Nicotiana tabacum and Its Functional Relation to Other Proteins

The trolC gene is a homologue of the rolC oncogene of Agrobacterium rhizogenes and most likely got into Nicotiana tabacum genome during horizontal gene transfer. This gene is quite conserved and is expressed in young organs, but its functions, like the agrobacterium rolC gene, remain unknown. The putative structure of the tRolC protein of Nicotiana tabacum, predicted using the I-TASSER service, was analyzed in the study. The presence of amino acid residues in the tRolC protein of the active center and binding site, as well as a protein-binding and ARTT loop, similar in structure to protein 6b of Agrobacterium vitis, indicates that they perform similar biological functions. Based on the data, a hypothesis was put forward about the presence of ADP-ribosyltransferase activity in the tRolC protein, which allows it to influence the growth and stress tolerance, as well as the biosynthesis of secondary metabolites by influencing the regulation of gene expression through interaction with transcription factors, RNA interference, or the manifestation of histone-like activity.

Genetically Encoded Microtubule Binders for Single-Cell Interrogation of Cytoskeleton Dynamics and Protein Activity.

Microtubule (MT) dynamics is tightly regulated by microtubule-associated proteins (MAPs) and various post-translational modifications (PTMs) of tubulin. Here, we introduce OligoMT and OligoTIP as genetically encoded oligomeric MT binders designed for real-time visualization and manipulation of MT behaviors within living cells. OligoMT acts as a reliable marker to label the MT cytoskeleton, while OligoTIP allows for live monitoring of the growing MT plus-ends. These engineered MT binders have been successfully utilized to label the MT network, monitor cell division, track MT plus-ends, and assess the effect of tubulin acetylation on the MT stability at the single-cell level. Moreover, OligoMT and OligoTIP can be repurposed as biosensors for quantitative assessment of drug actions and for reporting enzymatic activity. Overall, these engineered MT binders hold promise for advancing the mechanistic dissection of MT biology and have translational applications in cell-based high-throughput drug discovery efforts.

Effects of flavonoids on yeast ABC transporters activity

Flavonoids are known to be effective biomodulators of various processes in eukaryotic cells. As these compounds are present in wine and beer raw materials, they can influence the qualitative characteristics of the ethanol content in wine-making and brewing products, including directly through the mechanisms of regulation of gene expression and the activity of ATP-binding cassette (ABC) proteins. The main function of ABC transporters in yeast cells is to transport various substrates, including ethanol. This process ensures the survival of yeast cells under conditions of ethanol stress. It has been found that flavonoids, as well as their Schiff base derivatives, are effective stimulators and inhibitors of mRNA expression and activity of ABC proteins both in logarithmic and stationary phases of growth, which has a direct impact on bioethanol production by yeast.

Substituting ethoxyquin with tea polyphenols and propyl gallate enhanced feed oxidative stability, broiler hepatic antioxidant capacity and gut health

The safety of ethoxyquin has garnered increasing attention. This study evaluated the effects of partially substituting ethoxyquin with tea polyphenols and propyl gallate on feed oxidative stability, hepatic antioxidant properties, intestinal morphology and barrier functions, as well as the antioxidant and anti-inflammatory profiles of the intestinal mucosa in broilers. A total of 351 one-day-old male Arbor Acres Plus broilers were randomly assigned to 3 groups, each comprising 9 replicates with 13 birds per replicate. The treatments included a control group (CON) fed a basal diet, an ethoxyquin group (EQ) that received the basal diet supplemented with 120 g/t of ethoxyquin, and a substitution group (TP) receiving the basal diet supplemented with 6 g/t of tea polyphenols, 6 g/t of propyl gallate, and 30 g/t of ethoxyquin. In vitro experiments showed that both EQ and TP supplementation significantly reduced the acid value (AV), peroxide value (POV), and total oxidation value (TOV) of the feeds, with the TP group exhibiting lower AV and TOV than the EQ group. In vivo assessments revealed no significant differences in growth performance among the groups. Additionally, the TP group exhibited significantly higher glutathione peroxidase activity, increased glutathione content, and elevated protein expression of Keap1, Nrf2, and NQO1 in the liver compared to the control group (P < 0.05). Moreover, dietary TP significantly increased liver catalase activity, glutathione content, and NQO1 protein levels compared to the EQ group (P < 0.05). Both additives effectively reduced malondialdehyde levels in the intestinal mucosa by approximately 50% (P < 0.05) through the activation of the Nrf2/ARE pathway, as indicated by increased mRNA expression of TXN, CAT, GPX1, and GPX4 (P < 0.05). Furthermore, compared to the control group, the TP group exhibited greater villus height and villus height-to-crypt depth ratio (VCR) in the jejunum, as well as elevated VCR in the ileum (P < 0.05). The TP group also achieved the lowest serum levels of diamine oxidase activity, D-lactate and lipopolysaccharide contents among all groups (P < 0.05). The inclusion of both EQ and TP increased the mRNA expression of Occludin, Claudin-1, Mucin-2, and E-cadherin in the jejunum (P < 0.05). Moreover, the combination of tea polyphenols and propyl gallate effectively mitigated the proinflammatory effect of ethoxyquin, as evidenced by reductions in TNF-α, IL-18, and IFN-γ expression, potentially mediated by inhibition of the TLR-4/MyD88/NF-κB signaling pathway. In conclusion, this study demonstrates that partially replacing ethoxyquin with tea polyphenols and propyl gallate enhances feed oxidative stability, liver antioxidant capacity, and gut health in broilers, suggesting an efficient alternative with a lower dosage requirement.

The release behavior and in vitro osteogenesis of quercetin-loaded bioactive glass/hyaluronic acid/sodium alginate nanocomposite paste

Injectable pastes based on bioactive compounds and natural polymers are of interest in non-invasive bone surgeries. Several quantities of quercetin (100, 150, and 200 μM) were added to a sol-gel derived mesoporous bioactive glass. Injectable pastes based on quercetin-loaded bioactive glass, sodium alginate, and hyaluronic acid were prepared. Aggregated nanoparticles of bioactive glass and quercetin-loaded bioactive glass with mesoporous morphologies were confirmed by TEM and BET techniques. The quercetin release study was assessed in phosphate-buffered solution medium over 200 h and the obtained data were fitted by different eqs. A sustained release of quercetin was found, in which a better regression coefficient was achieved using Weibull equation. Human-derived mesenchymal stem cells were utilized to determine alkaline phosphatase activity and bone-related protein expression by western blotting and real-time PCR evaluations. Quercetin-loaded pastes increased the levels of alkaline phosphatase activity and the expression of Collagen-1, Osteopontin, Osteocalcin, and Runx2 proteins in a concentration-dependent manner. Due to the mesoporous architecture and high specific surface area of bioactive glass, the paste made of these particles and sodium alginate/hyaluronic acid macromolecules is appropriate matrix for quercetin release, resulting in promoted osteogenesis. The further in vivo studies can support the osteogenesis capacity of the quercetin-loaded paste.

ABA-regulated MAPK signaling pathway promotes hormesis in sugar beet under cadmium exposure

Sugar beet (Beta vulgaris L.) shows potential as an energy crop for cadmium (Cd) phytoremediation. To elucidate its in vivo response strategy to Cd exposure, seedlings were treated with 1, 3, and 5 mmol/L CdCl2 (Cd-1, Cd-3, and Cd-5) for 6 h, using 0 mmol/L CdCl2 (Cd-0) as the control. The results showed that Cd-3 promoted a unique “hormesis” effect, leading to superior growth performance, increased levels of chlorophyll, soluble protein, and SOD activity, and reduced MDA content in sugar beet, compared to Cd-1, Cd-5, and even Cd-0. GO and KEGG enrichments and PPI networks of transcriptomic analysis revealed that the differentially expressed genes (DEGs) were primarily involved in lipid metabolism, cellular protein catabolism, and photosynthesis. Notably, the MAPK signaling pathway was significantly enriched only under Cd-3, with the up-regulation of ABA-related core gene BvPYL9 and an increase in ABA content after 6 h of Cd exposure. Furthermore, overexpression of BvPYL9 in Arabidopsis thaliana (OE-1 and OE-2) resulted in enhanced growth (fresh weight, dry weight, and root length), as well as higher ABA and soluble protein contents under different Cd treatments. Cd-induced transcriptional responses of BvPYL9 were also evident in OE-1 and OE-2, especially at 10 µmol/L, indicated by qRT-PCR. These findings suggest that ABA-mediated MAPK signaling pathway is activated in response to Cd toxicity, with BvPYL9 being a key factor in the cascade effects for the Cd-induced hormesis in sugar beet.