Search Results (130)

Kefir is a probiotic beverage produced by symbiotic bacteria and yeasts. Polysaccharide-rich extracts from yellow and black soybeans (S and B) were obtained and subsequently fermented to produce S-F and B-F. The fermented extracts were further subjected to oxidative degradation using ascorbic acid and hydrogen peroxide to generate S-FD and B-FD. Physicochemical analyses revealed distinct differences in composition, phenolic profiles, and molecular weight among S-F, S-FD, B-F, and B-FD. Fourier transform infrared (FTIR) spectra indicated that oxidative degradation altered specific functional group intensities without disrupting the fundamental polysaccharide framework. Fermentation enhanced angiotensin-converting enzyme (ACE) inhibitory activity, and subsequent oxidative degradation further improved this effect. Both fermented and degraded extracts exhibited antioxidant activities, including 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity, ferrous-ion chelating ability, and reducing power, with degraded samples showing greater activity. The effects of the extracts on SH-SY5Y human neuroblastoma cells were evaluated in vitro. No cytotoxicity was observed at concentrations up to 400 μg/mL. Treatment at 200 μg/mL increased cell viability and reduced apoptosis in rotenone (ROT)-treated cells. Multivariate analysis further indicated that oxidative degradation enhanced antioxidant and ACE inhibitory activities but may reduce the protective effects observed in SH-SY5Y cells. Overall, soybean-derived kefir polysaccharide-rich extracts show potential as functional ingredients for applications related to blood pressure regulation and antioxidant activity, while their protective effects in neuronal cell models warrant further investigation. Full article

Bax, a key member of the B-cell lymphoma 2 (Bcl-2) protein family, is essential for inducing mitochondrial apoptosis. In this study, we employed yeast two-hybrid screening to identify ubiquitin-specific protease 49 (USP49) as a binding partner of Bax. Subsequent immunoprecipitation and glutathione S-transferase (GST) pull-down assays confirmed their direct interaction. Functional assays showed that USP49 reduces Bax polyubiquitination at multiple lysine residues within ubiquitin, with the strongest effects observed on K11, K29, K33, and K63 linkages. In contrast, its effect on K48-linked ubiquitination was weak and insufficient to influence Bax protein stability, indicating that USP49 does not regulate Bax abundance through proteasomal degradation. Instead, RT-qPCR analysis revealed that USP49 overexpression significantly increased Bax mRNA levels, and this effect was maintained under apoptosis stimuli (UV, H₂O₂, and STS), indicating transcriptional regulation largely independent of stress-induced damage, whereas its effect was modest and not statistically significant under starurosporine treatment. Collectively, these findings demonstrate that USP49 regulates Bax primarily through K29/K33/K63-linked ubiquitination and transcriptional upregulation, highlighting its role as a stress-responsive modulator of apoptosis and a potential therapeutic target in cancer. Moreover, under DNA damage condition (UV), USP49 overexpression marked enhanced apoptosis. Full article

Cancer remains a significant challenge in the field of medicine, primarily due to its inherent plasticity and the development of resistance to conventional therapeutic interventions. Genomic mutations and the activation of oncogenes enable cancer cells to resist senescence and apoptosis, leading to uncontrolled growth with harmful consequences. Small peptides are molecules with interesting anti-tumour properties and represent a valid alternative to conventional treatments. Our group has previously identified a class of small peptides bound to the DNA that can be extracted from the chromatin of various tissues, including wheat germ and trout. These peptide pools have been shown to possess interesting antiproliferative and apoptotic properties, and they are associated with cell cycle regulation. However, given the complexity of the extraction process, it is necessary to identify a substrate that will enable a more efficient extraction of these peptides, while also ensuring a composition that is simple to investigate. The present study developed a method for the extraction of this group of peptides from yeast, and the extract was then tested on cancer cells in order to confirm its anti-tumoral properties. The peptides were obtained from chromatin extracted from Saccharomyces cerevisiae cells through alkalisation and purification by gel filtration chromatography. The extract was tested on HeLa cells to verify its effects on vitality and the cell cycle. The data demonstrate that the chromatographic profile of this peptide extract indicates a more basic composition than the pool extracted from other tissues and exhibits comparable antiproliferative properties. The ability to rapidly obtain a biologically active, analytically accessible, and adequately purified fraction from the widely available substrate Saccharomyces cerevisiae represents a significant advance in the study of these DNA-binding peptides. Full article

Baker’s yeast beta-glucan (BYBG) supplementation improves various aspects of immune system function, readiness, and response. The purpose of this study was to determine if the expression of immune maturation mRNA was also changed over the course of 6 weeks of BYBG supplementation at rest. In this exploratory study, a small group of participants (N = 20) were randomized into two groups: BYBG (weeks 0–2 = 50 mg/d; 2–4 = 125 mg/d; and 4–6 = 250 mg/d) or placebo. Blood samples were collected at 0, 2, 4, and 6 weeks and analyzed for the expression of 785 mRNA (NanoString nCounter platform and Nanotube software; R v3.3.2). A total of 42 mRNAs in 21 annotated pathways (antigen presentation, apoptosis, B cell memory, cell cycle, chemokine signaling, cytotoxicity, DAP12 signaling, hypoxia response, IL-1 signaling, IL-10 signaling, MAPK signaling, myeloid immune response, NF-kB signaling, NK activity, Notch Signaling, PD1 signaling, Senescence/Quiescence, T cell checkpoint signaling, TCR signaling, TLR signaling, and TNF signaling), were significantly affected by BYBG at various time points. It is reasonable to speculate that the observed mRNA and associated pathways may underlie previously reported improvements in immune function with BYBG. Full article

The pro-apoptotic protein Bax is a key apoptosis regulator, as its activity is the main driver of mitochondrial outer membrane permeabilization. Bax is therefore tightly regulated, both by protein–protein interactions and post-translational modifications, such as phosphorylation. Although less studied, N-terminal acetylation has also been implicated in Bax regulation: disruption of the NatB N-terminal acetyl transferase complex in both yeast and MEFs increases Bax mitochondrial localization, although increased translocation is not sufficient to trigger its activation. Using the well-established model of heterologous expression of human Bax in yeast, we further investigated its regulation by N-terminal acetylation. We found that the sensitivity of Bax-expressing cells to acetic acid is greatly enhanced in a strain lacking the yeast NatB catalytic subunit (Nat3p). We propose that the Bax-induced cell death process shifts to a regulated necrosis in this strain due to autophagy inhibition. Furthermore, we show that the protective role of Bcl-xL against acetic acid-induced cell death of Bax-expressing yeast cells requires Nat3p. We speculate that Nat3p modulates the function of pro-death and pro-survival proteins, ultimately affecting both the levels and mode of cell death. These findings may have implications for the development of novel therapeutic strategies targeting human diseases associated with cell death dysfunction. Full article

Background/Objectives: Mutations in NACHT, LRR and PYD domain-containing protein 2 (NLRP2) and NLRP7 genes, members of the NOD-like receptor (NLR) family of innate immune sensors, result in recurrent miscarriages and reproductive wastage in women. These genes have been identified to be maternal effect genes in humans and mice regulating early embryo development. Previous research in vitro suggests that NLRP2 and NLRP7 regulate DNA methylation and/or immune signaling through inflammasome formation. However, the exact mechanisms underlying NLRP2 and NLRP7 function are not well defined. Methods: To determine the interacting proteins required for NLRP2/NLRP7-mediated regulation of DNA methylation, yeast 2-hybrid screens, coimmunoprecipitation, and FRET studies were performed and verified the ability of novel protein interactions to affect global DNA methylation by 5-methylcytosine-specific ELISA. Results: Various methodologies employed in this research demonstrate a novel protein interaction between human ErbB3-binding protein 1 (EBP1, also known as proliferation-associated protein 2G4 (PA2G4) and NLRP2 or NLRP7. In addition, NLRP2 and NLRP7 regulate EBP1 gene expression. Functionally, global DNA methylation levels appeared to decrease further when NLRP2 and NLRP7 were co-expressed with EBP1, although additional studies may need to confirm the significance of this effect. Conclusions: Since EBP1 is implicated in apoptosis, cell proliferation, DNA methylation, and differentiation, our discovery significantly advances our understanding of how mutations in NLRP2 or NLRP7 may contribute to reproductive wastage in women through EBP1. Full article

This review highlights the emerging functional implications of nonsense-mediated mRNA decay (NMD) in human diseases, with a focus on its therapeutic potential for cardiovascular disease. NMD, conserved from yeast to humans, is involved in apoptosis, autophagy, cellular differentiation, and gene expression regulation. NMD is a highly conserved surveillance mechanism that degrades mRNAs containing premature termination codons (PTCs) located upstream of the final exon-exon junction. NMD serves to prevent the translation of aberrant mRNA and prevents the formation of defective protein products that could result in diseases. Key players in this pathway include up-frameshift proteins (UPFs), nonsense-mediated mRNA decay associated with p13K-related kinases (SMGs), and eukaryotic release factors (eRFs), among others. Dysregulation of NMD has been linked to numerous pathological conditions such as dilated cardiomyopathy, cancer, viral infections, and various neurodevelopmental and genetic disorders. This review will examine the regulatory mechanisms by which NMD regulation or dysregulation may contribute to disease mitigation or progression and its potential for cardiovascular disease therapy. We will further explore how modulating NMD could prevent the outcomes of mutations underlying genetically induced cardiovascular conditions and its applications in personalized medicine due to its role in gene regulation. While recent advances have provided valuable insights into NMD machinery and its therapeutic potential, further studies are needed to clarify the precise roles of key NMD components in cardiovascular disease prevention and treatment. Full article

As a pivotal model organism in Lepidoptera research, the silkworm (Bombyx mori) holds significant importance in life science due to its economic value and biotechnological applications. Advancements in proteomics and bioinformatics have enabled substantial progress in characterizing the B. mori proteome. Systematic screening and identification of protein–protein interactions (PPIs) have progressively elucidated the molecular mechanisms governing key biological processes, including viral infection, immune regulation, and growth development. This review comprehensively summarizes traditional PPI detection techniques, such as yeast two-hybrid (Y2H) and immunoprecipitation (IP), alongside emerging methodologies such as mass spectrometry-based interactomics and artificial intelligence (AI)-driven PPI prediction. We critically analyze the strengths, limitations, and technological integration strategies for each approach, highlighting current field challenges. Furthermore, we elaborate on the molecular regulatory networks of Bombyx mori nucleopolyhedrovirus (BmNPV) from multiple perspectives: apoptosis and cell cycle regulation; viral protein invasion and trafficking; non-coding RNA-mediated modulation; metabolic reprogramming; and host immune evasion. These insights reveal the dynamic interplay between viral replication and host defense mechanisms. Collectively, this synthesis aims to provide a robust theoretical foundation and technical guidance for silkworm genetic improvement, infectious disease management, and the advancement of related biotechnological applications. Full article

ADF/cofilins form a family of small, widely expressed actin-binding proteins, regulating actin dynamics in various cellular and physiological processes in all eukaryotes, from yeasts to animals. Changes in the expression of the ADF/cofilin family proteins have been demonstrated under various pathological conditions. The well-established role of cofilin in migration, invasion, epithelial-mesenchymal transition, apoptosis, resistance to radiotherapy and chemotherapy, immune escape, and transcriptional dysregulation in malignant tumors is primarily attributed to its actin-modifying activity. Moreover, drugs targeting this function of cofilin have been developed for cancer treatment. However, its multilevel regulation, highly diverse effects across various pathological conditions, and conflicting data on the functional consequences of altered cofilin expression have prompted us to explore additional roles of cofilin—beyond actin modulation—particularly its involvement in lipid metabolism and mitochondrial homeostasis. Here, we review recent data on the expression of ADF/cofilin family proteins in various pathologies, account for the mutations and post-translational modifications of these proteins and their functional consequences, dwell on the role of K63-type ubiquitination of cofilin for its involvement in lipid metabolism and mitochondrial homeostasis, more specifically, a process of mitochondrial division or mitofission, point out conflicting data in cofilin research, and describe prospects for future studies of cofilin functions. Full article

Shinorine is a mycosporine-like amino acid isolated from laver (Porphyra dentata), and interest in its functionality has increased recently due to increased production using yeast. There have been few reports on the pharmacological activity of shinorine, and we sought to find the pharmacological significance of shinorine. In the present study, we investigated the pharmacological effects of shinorine purified from Porphyra dentata on ARPE-19 cells. First, when ARPE-19 cells were treated with bis-retinoid N-retinyl-N-retinylidene ethanolamine (A2E) and blue light (BL), cytotoxicity increased, and apoptosis was observed. We investigated the effects of shinorine on A2E- and BL-induced cytotoxicity and changes in apoptotic factors, inflammation, and carbonyl stress. A2E and BL exposure increased ARPE-19 cell apoptosis, but this increase was attenuated by shinorine in a concentration-dependent manner. Treatment with A2E and BL induced ARPE-19 cell apoptosis, but treatment with shinorine decreased the apoptotic factors, such as MAPKs. Shinorine reduced p-JNK and p-P38, which were increased by A2E and BL. In addition, shinorine was found to regulate inflammatory proteins and proteins associated with carbonyl stress. In conclusion, shinorine may suppress cell damage caused by A2E treatment and BL exposure at the cellular level by regulating various cell death and inflammatory response pathways. Full article

The aging population is steadily increasing, with aging and age-related diseases serving as major risk factors for morbidity, mortality, and economic burden. Peanuts, known as the “longevity nut” in China, have been shown to offer various health benefits, with peanut skin extract (PSE) emerging as a key compound of interest. This study investigates the bioactive compound in PSE with anti-aging potential and explores its underlying mechanisms of action. Procyanidin A1 (PC A1) was isolated from PSE, guided by the K6001 yeast replicative lifespan model. PC A1 prolonged the replicative lifespan of yeast and the yeast-like chronological lifespan of PC12 cells. To further confirm its anti-aging effect, cellular senescence, a hallmark of aging, was assessed. In senescent cells induced by etoposide (Etop), PC A1 alleviated senescence by reducing ROS levels, decreasing the percentage of senescent cells, and restoring proliferative capacity. Transcriptomics analysis revealed that PC A1 induced apoptosis, reduced senescence-associated secretory phenotype (SASP) factors, and modulated the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. The antioxidative capacity of PC A1 was also evaluated, showing enhanced resistance to oxidative stress in PC12 cells by reducing reactive oxygen species (ROS) and malondialdehyde (MDA) levels and increasing superoxide dismutase (SOD) activity. Moreover, PC A1 induced autophagy, as evidenced by an increase in fluorescence-labeled autophagic compartments and confirmation via Western blot analysis of autophagy-related proteins. In addition, the treatment of an autophagy inhibitor abolished the antioxidative stress and senescence-alleviating effects of PC A1. These findings reveal that PC A1 extended lifespans and alleviated cellular senescence by enhancing oxidative stress resistance and inducing autophagy, positioning it as a promising candidate for further exploration as a geroprotective agent. Full article

Salvia apiana Jepson is an endemic North American species characterized by a rich phytochemical profile including abietane-type diterpenoids, phenolic acids, flavonoids, and thujone-free essential oil (EO). The current study was aimed at increasing EO production in bioreactor-grown S. apiana microshoot culture through biotic elicitation using chitosan, ergosterol, and yeast extract (YE). Additionally, the immunomodulatory effects of the major volatile constituent of white sage—1,8-cineole—as well as EOs obtained from both S. apiana microshoots and leaves of field-grown plants, were assessed. EOs were isolated via hydrodistillation and analyzed by GC/MS and GC/FID. Biological assays included flow cytometric evaluation of the proliferation and apoptosis rates of human CD4 and CD8 T lymphocytes, obtained from healthy volunteers and subjected to different concentrations of EOs and 1,8-cineole. Elicitation with 100 mg/L YE improved the production of EO in S. apiana microshoots by 9.4% (1.20% v/m). EOs from both microshoots and leaves of field-grown plants, as well as 1,8-cineole, demonstrated dose-dependent anti-proliferative and pro-apoptotic effects on CD4+ and CD8+ T cells. These findings highlight the potential of S. apiana microshoot cultures capable of producing EO with significant immunomodulatory activity. Full article

Background: A comparative 12-week dietary intervention of red yeast rice (RYR, Beni-koji, Kobayashi, Japan) and Cuban policosanol (PCO, Raydel^®, Thornleigh, Australia) was assessed for dyslipidemia, antioxidant status, and vital organ functionality in hyperlipidemic zebrafish. Methods: Hyperlipidemic zebrafish were supplemented with a high-cholesterol diet (HC, final 4%, w/w) infused with either a powdered RYR tablet (final 1.0%, w/w), a PCO tablet (final 1.0%, w/w), or a combination of 0.5% (w/w) each of RYR and PCO powder for 12 weeks. Subsequently, blood and organs were collected and processed for biochemical and histological examination. Results: RYR and PCO consumption showed a substantial effect against HC-induced hyperlipidemia by reducing the total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C). Exclusively, PCO supplementation revealed a significant elevation in the HC-diminished high-density lipoprotein cholesterol (HDL-C). In addition, PCO supplementation showed a significant elevation in plasma ferric ion-reducing ability (FRA) and sulfhydryl content, as well as alleviating the blood glucose level of hyperlipidemic zebrafish. The most noteworthy impact, with a significant two-fold (p < 0.001) augmentation of HC-diminished plasma paraoxonase (PON) activity, was observed in response to PCO. In contrast, the RYR supplementation failed to establish curative effects against HC-disturbed plasma antioxidant variables and blood glucose levels. The histological outcome revealed a severe toxicological impact of the RYR on the liver, reflected by fatty liver changes and three-fold heightened IL-6 production compared to HC control. Contrastingly, PCO exhibited significant hepatoprotection and effectively neutralized the hepatic toxicity triggered by HC and RYR. Also, RYR showed kidney atrophy, intense ROS generation, apoptosis, and senescence. Conversely, the PCO supplementation protected the kidney from HC- and RYR-induced toxicity. Likewise, PCO supplementation notably alleviated histological alterations and oxidative stress in the brain, ovary, and testis of hyperlipidemic zebrafish. Conclusions: This comparative study establishes PCO’s therapeutic effect against the challenges posed by HC, while RYR emerged with serious toxicological concerns towards the liver, kidney, and other organs of hyperlipidemic zebrafish. Full article

Wine lees, the second most significant by-product of winemaking after grape pomace, have received relatively little attention regarding their potential for valorization. Despite their rich content in bioactive components such as β-glucans, industrial utilization faces challenges, particularly due to variability in their composition. This inconsistency impacts the reliability and standardization of final products, limiting broader adoption in industrial applications. β-Glucans are dietary fibers or polysaccharides renowned for their diverse bioactive properties, including immunomodulatory, antioxidant, anti-inflammatory, antitumor, and cholesterol- and glucose-lowering effects. They modulate the immune system by activating Dectin-1 and TLR receptors on immune cells, enhancing phagocytosis, cytokine production, and adaptive immune responses. Their antioxidant activity arises from neutralizing free radicals and reducing oxidative stress, thereby protecting cells and tissues. β-Glucans also exhibit antitumor effects by inhibiting cancer cell growth, inducing apoptosis, and preventing angiogenesis, the formation of new blood vessels essential for tumor development. Additionally, they lower cholesterol and glucose levels by forming a viscous gel in the intestine, which reduces lipid and carbohydrate absorption, improving metabolic health. The biological activity of β-glucans varies with their molecular weight and source, further highlighting their versatility and functional potential. This study investigates how grape variety, vinification technology and extraction methods affect the yield and properties of β-glucans extracted from wine lees. The physico-chemical and mineral composition of different wine lees were analyzed, and two extraction methods of β-glucans from wine lees were tested: acid-base extraction and autolysis. These two methods were also tested under ultrasound-assisted conditions at different frequencies, as well as without the use of ultrasound. The β-glucan yield and properties were evaluated under different conditions. FTIR spectroscopy was used to assess the functional groups and structural characteristics of the β-glucans extracted from the wine lees, helping to confirm their composition and quality. Rheological behavior of the extracted β-glucans was also assessed to understand the impact of extraction method and raw material origin. The findings highlight that vinification technology significantly affects the composition of wine lees, while both the extraction method and yeast origin influence the yield and type of β-glucans obtained. The autolysis method provided higher β-glucan yields (18.95 ± 0.49% to 39.36 ± 0.19%) compared to the acid–base method (3.47 ± 0.66% to 19.76 ± 0.58%). FTIR spectroscopy revealed that the β-glucan extracts contain a variety of glucan and polysaccharide types, with distinct β-glucans (β-1,4, β-1,3, and β-1,6) identified through specific absorption peaks. The rheological behavior of suspensions exhibited pseudoplastic or shear-thinning behavior, where viscosity decreased significantly as shear rate increased. This behavior, observed across all β-glucan extracts, is typical of polymer-containing suspensions. These insights are critical for optimizing β-glucan extraction processes, supporting sustainability efforts and waste valorization in the wine industry. Efficient extraction of β-glucans from natural sources like wine lees offers a promising path toward their industrial application as valuable functional compounds. Full article