Biology and Life Sciences

Abstract: Anthraquinone-based intercalating compounds, such as doxorubicin and mitoxantrone, have long been used clinically due to their ability to induce DNA damage. More recently, heteroarene-fused anthraquinones have been developed to further enhance their anticancer activity. Among these compounds, 4,11-bis(2-(2-chloroacetamidine)ethylamino)anthra[2,3-b]thiophene-5,10-dione dihydrochloride (designated as derivative a) was identified as a potent apoptotic inducer. Based on this scaffold, two additional derivatives were synthesized by replacing the sulfur atom within the heterocyclic ring with nitrogen (derivative b) or oxygen (derivative c). Building upon our previous identification of ENOX2 as the primary target of this scaffold, the present study investigated the anti-proliferative effects and underlying mechanisms of these derivatives in colon cancer cells with varying p53 statuses. Derivatives a and b effectively induced apoptosis and suppressed proliferation in p53 wild-type HCT116 cells, which was concomitantly accompanied by significant ENOX2 downregulation and the activation of intrinsic apoptotic signaling. In contrast, p53-null HCT116 cells exhibited reduced sensitivity, attenuated apoptotic responses, and minimal ENOX2 downregulation. Notably, derivative c primarily induced G2/M arrest rather than apoptosis regardless of p53 status, indicating a predominantly cytostatic mechanism. Collectively, these findings suggest that the degree of ENOX2 modulation is linked to the distinct anti-proliferative responses induced by heteroarene-fused anthraquinones, and that p53 status serves as a critical molecular switch influencing the transition between cytostatic growth arrest and apoptotic cell death.

Abstract: Metabolites are commonly interpreted through abundance, pathway flux, pathway assignment, or thermodynamic feasibility. These measurements are essential, but they do not by themselves explain how the same molecule is assigned to different biochemical functions. Here we develop the partition theory of metabolite function to describe this allocation explicitly. We define the partition vector, π(τ), for a metabolite M as the central variable of the framework. Its components represent the time-integrated fractions of M captured by competing reaction, pathway, or compartmental sinks during a biologically relevant decision window τ. Because π(τ) is normalized, it describes how metabolite utilization is distributed among competing fates, rather than how much metabolite is present or how much total flux passes through a pathway. This framework leads naturally to partition entropy as a measure of metabolite-fate ambiguity, to partition-control relations that distinguish flux-changing from fate-redirecting perturbations, to cross-metabolite coupling through shared sinks, and to history dependence when downstream marks or assembly states retain the record of metabolite use. The framework does not replace metabolic control analysis, flux balance analysis, or thermodynamic flux analysis. Rather, it reorganizes quantities that these approaches already help define into a metabolite-centered state variable, the fraction of utilization captured by each competing sink. A literature-constrained acetyl-CoA example illustrates how sink sensitivity can be evaluated without inventing a universal empirical acetyl-CoA partition vector.

Abstract: Background. Metadichol® (Nano-Policosanol; Nanorx Inc.) is a nano-emulsion of food-derived very-long-chain primary alcohols (C26–C36) that acts as a Vitamin D Receptor (VDR) inverse agonist at picomolar to nanomolar concentrations. Prior peer-reviewed work has documented its regulation of all 49 nuclear receptors, all 7 sirtuins, Toll-like receptors, and the Yamanaka pluripotency factors. The 60S large ribosomal subunit—the catalytic engine of the ribosome housing the peptidyl-transferase centre—is encoded by 47 ribosomal protein genes (RPL/RPLP) that also perform extraribosomal roles in p53 surveillance, inflammatory mRNA silencing, and developmental patterning. Whether a single food-derived compound can coordinately regulate this entire repertoire has not been tested. Methods. Human PBMCs were isolated by Histopaque-1077 density-gradient centrifugation and treated for 24 h with Metadichol at 0.1 pg/mL, 1 pg/mL, 100 pg/mL, 1 ng/mL, and 100 ng/mL alongside untreated controls. Total RNA was extracted (TRIzol), reverse-transcribed (500 ng; PrimeScript), and all 47 large-subunit RPL/RPLP genes were quantified by SYBR-Green qRT-PCR (39 cycles; 60 °C annealing) using gene-specific validated primers. Relative expression was computed by the 2^−ΔΔCq method with GAPDH as reference; significance by one-way ANOVA with Dunnett's test. Results. All 47 large-subunit ribosomal protein genes were significantly up-regulated by Metadichol relative to untreated control, but in a strikingly coherent, non-monotonic (biphasic) manner. At the lowest dose (0.1 pg/mL) 44 of 47 genes were suppressed below control (mean 0.53-fold), whereas activation rose to a sharp maximum at 1 ng/mL—where 33 of 47 genes reached their individual peak (mean 2.27-fold)—before partial relaxation at 100 ng/mL. The largest single responses were RPL30 (5.43-fold), RPL24 (4.78-fold), RPL37 (4.31-fold), RPL38 (4.30-fold), RPL34 (4.17-fold), RPL18 (3.92-fold), RPL37A (3.83-fold), and RPL23A (3.80-fold). The p53-axis regulators RPL5, RPL11, and RPL26, the GAIT-complex protein RPL13A, the Hox-selective translator RPL38, and the P-stalk genes RPLP0/RPLP1/RPLP2 were all induced. Hierarchical clustering resolved a dominant "1 ng/mL-peaking" co-regulated module; inter-dose correlation analysis showed the 1 ng/mL and 100 pg/mL profiles were the most concordant (r ≈ 0.52) while the suppressive 0.1 pg/mL profile was weakly correlated with the activating doses (r ≈ 0.25), defining two regulatory regimes. A 47×47 gene–gene correlation matrix revealed broadly positive co-regulation, consistent with a single shared upstream driver rather than gene-by-gene noise. Conclusions. Metadichol produces a stoichiometrically balanced, dose-tuned activation of the entire 60S ribosomal protein gene set in primary human immune cells. To our knowledge this is the first agent shown to coordinately up-regulate the complete large-subunit repertoire, and it runs counter to the existing ribosome pharmacopeia, which is almost entirely inhibitory (mTOR inhibitors, RNA Pol I inhibitors). The uniform direction and tight gene–gene correlation distinguish this from the selective, imbalanced RPL changes that drive ribosomopathies (Diamond–Blackfan anemia, 5q- MDS, T-ALL), and instead resemble the coordinated ribosome-biogenesis program of pluripotent and regenerating cells. A notable RPL5↔RPL11 (5S RNP) asymmetry marks RPL5 as the single most dose-sensitive node. These findings position Metadichol as the first coordinated, balanced transcriptional activator of translational capacity—the directional mirror image, at the level of gene expression, of both ribosomopathy and ribosome-inhibitor drugs—with implications for ribosomopathy, iPSC reprogramming, immune function, and ageing biology.

Abstract: Adiponectin is an important metabolic biomarker associated with insulin sensitivity, obesity, type 2 diabetes, and cardiovascular risk, but routine adiponectin measurement still depends mainly on centralized laboratory platforms. To provide a practical method for rapid and decentralized serum testing, we developed a smartphone-based quantitative lateral flow immunoassay (LFIA) for adiponectin and evaluated its analytical performance. The platform integrates a colorimetric gold nanoparticle-based LFIA strip, a smartphone reader equipped with macro-optics and controlled illumination, and a dedicated Android application for standardized image acquisition and quantitative grayscale analysis. Following systematic optimization of critical assay parameters, the developed method achieved a quantitative range of 0.32-40 mg/L, with limits of detection (LOD) and quantification (LOQ) of 0.16 and 0.32 mg/L, respectively. Analytical evaluation demonstrated good repeatability and intermediate precision, with within-run coefficients of variation of 2.40-4.66% and between-run coefficients of variation of 3.31-6.12%. Interference testing indicated borderline interference performance, with bilirubin, triglycerides, and hemoglobin recoveries slightly above the predefined acceptance limit and rheumatoid factor at the lower acceptance boundary under the evaluated moderate-concentration condition. Accelerated stability testing showed that the strips retained acceptable analytical responses after storage at 50 °C for 28 days. Method comparison using 125 clinical serum samples demonstrated close agreement with a commercial particle-enhanced turbidimetric immunoassay (PETIA) comparator method, with good linear association by ordinary regression (R² = 0.9688) and minimal mean bias by Bland-Altman analysis (-0.0392 mg/L; 95% limits of agreement: -1.1601 to 1.0817 mg/L). These results indicate that the proposed smartphone-assisted LFIA is an operationally simple method for quantitative serum adiponectin determination and may support point-of-care or near-patient metabolic biomarker monitoring.

Abstract: Modern proteomics faces a critical bottleneck: the vast discrepancy between the number of genes in the human genome and the exponentially greater variety of functional proteoforms that actually drive biological processes. Our paper addresses the urgent need for high-resolution systematic mapping of these proteoforms, arguing that the true frontier of molecular biology lies in the precise identification and categorization of protein variants. It centers on the development and expansion of the "2DE-pattern" database, a specialized platform designed to bridge the gap between theoretical protein sequences and the physical reality of protein expression as captured through two-dimensional electrophoresis (2DE). A database “2DE-pattern”is based on information obtained by separation of proteoforms using 2DE with following shot-gun ESI LC-MS/MS. It was launched in 2020, contains multiple isoform-centric patterns of proteoforms, and can be freely used at http://2de-pattern.pnpi.nrcki.ru. Here, we report the additional data and all updates that were added into this database. Also, it was upgraded to be more research-oriented. Some tools were incorporated into the database to allow the convenient comparative analysis of the data. Accordingly, it became now more like a knowledge database.

Abstract: One-carbon (1C) metabolism involves the transfer of a methyl group from one molecule to another, to effect crucial cellular functions, including DNA, RNA, and protein methylation, as well as synthesis of phosphatidylcholine and L-carnitine. In this paper, we argue that impaired 1C metabolism plays a central role in mitochondrial dysfunction, due to deuterium overload in mitochondrial F1F0 ATP synthase (ATPase). Deuterium, a heavy isotope of hydrogen, damages ATPase, causing inefficiencies in ATP production and increased release of reactive oxygen species. We argue that gut microbes play a crucial role in assuring that 1C units are virtually deuterium free. S-adenosylmethionine serves as the universal methyl donor. We hypothesize that gut microbes produce extremely deuterium depleted (deupleted) hydrogen gas which they use as a reducing agent to convert carbon dioxide into organic molecules, including acetate, butyrate, formate, and the methyl groups carried in 1C metabolism. The methyl group is sourced from methyl-tetrahydrofolate (CH3-THF), while methionine, glycine, serine, formaldehyde, formate, choline, L-carnitine, and melatonin are carriers of 1C units. Through an in-depth review of methylation and demethylation processes, we hypothesize that methyl groups ultimately deliver deupleted protons to the mitochondria. Deficiencies in nutrients carrying 1C units, disrupt methylation pathways, causing disease through co-causality of mitochondrial dysfunction. Synthetic versions of choline, L-carnitine, and methionine may be problematic because they supply methyl groups that are not deupleted. Trimethylamine oxide (TMAO), derived from microbial metabolism of choline and L-carnitine, followed by oxidation in the liver, is a marker for fatty liver disease and cardiovascular disease, and it may serve as a signaling molecule for both gut dysbiosis and deuterium overload.

Abstract: Melanin biosynthesis proceeds through dopaquinone, a reactive ortho-quinone that partitions between eumelanin and pheomelanin depending on local thiol availability. When intracellular cysteine and glutathione are sufficient, dopaquinone is intercepted to form cysteinyldopa and ultimately pheomelanin. When thiols are depleted, dopaquinone undergoes intramolecular cyclization toward eumelanin through dopachrome, DHI, and DHICA. This much is well established. What has received less attention is the consequence that follows: eumelanin synthesis itself consumes thiols, generates quinonoid intermediates with residual electrophilic reactivity, and sustains a pro-oxidant microenvironment within the melanosome and its surroundings. The resulting oxidative burden further depletes the thiol pool, which in turn shifts subsequent dopaquinone partitioning still further toward eumelanin. We term this self-reinforcing sequence the Melanin Loop—a positive-feedback cycle in which eumelanin production creates the chemical conditions that favor yet more eumelanin production. An analogous electrophilic burden operates in photoaging. Ultraviolet radiation drives lipid peroxidation, yielding 4-hydroxynonenal (4-HNE) and acrolein—reactive carbonyl species that form covalent adducts with dermal collagen and elastin. Both the melanin loop and UV-driven carbonyl stress share a common chemical feature: nucleophilic cellular defenses are overwhelmed by electrophilic intermediates that conventional antioxidants do not efficiently neutralize. We propose that Passive Electron Donors (PEDs)—compounds capable of directly quenching electrophilic intermediates downstream of dopaquinone and downstream of lipid peroxidation—represent a rational third layer of intervention, complementing sunscreens (photon blockade) and classical antioxidants/thiols (redox buffering). This paper presents the mechanistic framework and outlines a translational research agenda; it does not claim clinical proof for any specific PED formulation.

Abstract: Nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H₂S) were once regarded solely as toxic environmental gases. However, accumulating evidence over the past several decades has established them as the three principal endogenous gasotransmitters that regulate a wide spectrum of physiological and pathological processes. Unlike conventional signaling molecules, gasotransmitters diffuse freely across biological membranes and exert potent biological effects through receptor-independent mechanisms, including redox-sensitive post-translational modifications and modulation of heme-containing proteins. Although the individual functions of NO, CO, and H₂S have been extensively reviewed, emerging studies indicate that these gaseous mediators rarely operate in isolation. Instead, they form a highly integrated signaling network characterized by direct chemical interactions, reciprocal enzymatic regulation, and convergence upon common downstream pathways. In this mini-review, we propose the concept of a “Gasotransmitter Trio Network,” emphasizing the molecular crosstalk among NO, CO, and H₂S as a fundamental determinant of cellular homeostasis. We first summarize the biosynthetic pathways and major signaling mechanisms of the gasotransmitter trio, including S-nitrosylation, persulfidation, and heme-dependent regulation. We then discuss recent advances revealing how interactions among these gases generate novel bioactive intermediates and coordinate redox signaling. Particular attention is given to the emerging roles of gasotransmitters in regulating ferroptosis, autophagy, and mitophagy by modulating iron metabolism, lipid peroxidation, mitochondrial quality control, and antioxidant defense systems. These findings support a unified framework in which gasotransmitters function as master regulators of cellular fate under conditions of physiological and pathological stress. Finally, we highlight recent progress in stimuli-responsive donors, carbon monoxide-releasing molecules (CORMs), nitric oxide-releasing materials (NORMs), hydrogen sulfide donors, and advanced nanoplatforms that enable spatiotemporally controlled gas delivery. We propose that future therapeutic strategies will increasingly rely on programmable multi-gas systems that recapitulate endogenous gasotransmitter networks. Collectively, this review provides a systems-level perspective on gasotransmitter biology and outlines emerging opportunities for the development of precision gas medicine in cardiovascular, neurodegenerative, inflammatory, metabolic, and malignant diseases.

Abstract: Background & Objectives: The increasing prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) is intrinsically linked to nutrient overload and obesity, and adipose dysfunction. While continuous dietary stress triggers adipose-tissue-derived lipotoxicity and disrupts hepatic metabolic homeostasis and provokes inflammation, the transcriptional scaffolds that mitigate this lipotoxicity remain incompletely understood. We investigated the role of zinc finger protein 90 (ZFP90) in defending against diet-induced metabolic stress and MASLD pathogenesis. Methods: Wild-type and ZFP90-knockout mice were subjected to a high-fat diet (HFD) to model nutrient-overload-induced MASLD. Hepatic phenotypes were characterized using metabolic profiling and RNA sequencing. Mechanistic dynamics were evaluated through protein interaction assays, and clinical relevance was validated using human MASLD liver biopsies. Results: ZFP90 deficiency significantly accelerated HFD-induced steatosis, systemic insulin resistance, and inflammatory infiltration. Crucially, ZFP90 depletion drove severe white adipose tissue (WAT) dysfunction, characterized by impaired lipogenic capacity, exacerbated lipolysis, and diminished local insulin signaling. This was accompanied by a pro-inflammatory secretory shift in WAT, evident from decreased Adipoq and increased Cd68/Ccl3 expression. In the liver, transcriptomic analysis revealed a profound induction of pathways related to fatty acid uptake and cytokine signaling. Mechanistically, ZFP90 forms a repressive complex with TRIM28, acting as a crucial molecular brake on NF-κB signaling. Loss of ZFP90 unleashes p65-mediated hyper-inflammation. Clinically, hepatic ZFP90 expression is significantly upregulated in patients with MASLD. Conclusions: ZFP90 is a novel regulator of immunometabolic homeostasis under dietary stress. By forming of complex with Trim28 to inhibit the nuclear translocation of NF-κB, ZFP90 suppresses pro-inflammatory responses and protects the liver from obesity-associated systemic lipotoxicity. These finding provide critical insights into the adipo-hepatic axis and highlight the ZFP90 as a promising therapeutic target to mitigate the progression to metabolic dysfunction-associated steatohepatitis (MASH).

Abstract: Milk-derived extracellular vesicles (EVs) transport microRNAs (miRNAs) that are un-usually stable and have been proposed to survive digestion and modulate gene expres-sion in the consumer, although their dietary bioavailability and physiological rele-vance remain debated. How the predicted regulatory potential of these miRNAs differs among the milks most relevant to human nutrition has not been systematically com-pared. Here we performed an integrative in silico analysis of publicly available small-RNA sequencing data from 29 milk and milk-cell samples of human, cow, goat and donkey origin. miRNAs were quantified against human (hsa) miRBase refer-ences—thereby restricting the analysis to evolutionarily conserved miRNAs with hu-man orthologs—and their predicted effect on the human transcriptome was modeled by integrating predicted (mirDIP) and experimentally supported (TarBase v9) miR-NA–target interactions into a per-gene, per-species weighted targeting score. Because miRNAs act predominantly as repressors, this score is read as a prediction of which genes would be post-transcriptionally down-regulated in a recipient. miR-148a-3p dominated the exosomal spectrum of all four species (≈21.5% of pooled abundance), and the twenty most abundant miRNAs accounted for roughly three quarters of the signal. Of 4,577 robustly targeted genes, a 1,809-gene conserved “pan-milk” core showed the highest cross-species targeting and was enriched for transcriptional regu-lation, PI3K–Akt, MAPK and TGF-β/SMAD signaling, autophagy and—strikingly—the components of the RNA-interference machinery itself. Species-restricted gene sets re-capitulated biologically plausible programs, including a human-biased neu-ronal/axon-guidance and chromatin module, a donkey-biased transcriptional, epithe-lial and immune (CD47) module, and a ruminant lipid/cholesterol and insulin–mTOR module. Across categories we observed a reproducible confidence–exclusivity trade-off. We emphasize that these results are computational predictions that assume dietary miRNA uptake and do not constitute experimental validation. We provide the complete targetome (Supplementary Materials S1–S9) as a hypothesis-generating re-source to prioritize candidate genes, pathways and milk types for future functional, nutritional and epigenetic investigation.

Abstract: Adipose tissue has emerged as a pivotal endocrine organ, secreting bioactive proteins termed adipokines that regulate metabolic and immune processes across multiple organ systems. In the context of sepsis and critical illness, conditions defined by a dysregulated host response to infection with life-threatening organ dysfunction, the role of novel adipokines has attracted considerable research interest. This review focuses on three novel adipokines, chemerin, vaspin (SERPINA12), and omentin-1 (intelectin-1). We will discuss current in vitro, in vivo experimental animal models, and clinical evidence, emphasizing their biology, mechanisms of action, and potential as diagnostic and prognostic biomarkers in critically ill patients. All three adipokines are elevated in sepsis compared with healthy controls and correlate with established severity scores, including APACHE II and SOFA. Chemerin and omentin-1 have both been independently associated with 28-day mortality in prospective cohort studies. Vaspin exhibits robust cardioprotective effects in murine sepsis models via inhibition of kallikrein 7 (KLK7) and attenuates lipopolysaccharide (LPS)-induced acute lung injury (ALI) both in vitro and in vivo. Omentin-1 suppresses LPS-induced macrophage activation through TLR4/MyD88/NF-κB inhibition in vitro and protects against LPS-induced ALI in murine models. Despite these promising findings, substantial methodological heterogeneity and limited large-scale clinical data currently preclude clinical implementation. Future research that standardizes assays, expands to multicenter cohorts, and investigates therapeutic modulation of these pathways is urgently needed.

Abstract: Personalized oncology seeks to selectively block specific dysregulated pathways to ar-rest cancer development. Increased metabolism of glutamine is a hallmark of cancer, and 6-diazo-5-oxo-L-norleucine (DON), a structural analog of L-glutamine, was the first compound aimed to target the exacerbated nitrogen metabolism observed in can-cer cells. However, it was abandoned due to unacceptable side toxicity. With the same goal of blocking glutamine metabolism, several specific glutaminase inhibitors have been characterized in recent decades, showing promising results. Nevertheless, this strategy frequently induces adaptive metabolic resistance that must be counteracted. In this context, glutaminase has become a key target in combination therapies for sev-eral tumor types aimed at restricting anabolic adaptation when single metabolic ther-apy fails, emerging as a possible synergistic therapeutic intervention. Consequently, combination therapies that include glutaminase inhibition alongside additional drug/s to counteract the metabolic plasticity of cancer have become essential in antitumor personalized pharmacology. Recent findings suggest that novel prodrugs targeting glutamine metabolism can potentiate immune checkpoint inhibitors by reshaping the tumor microenvironment, thereby enhancing cancer immunotherapy while reducing side effects and increasing therapeutic efficacy in certain types of cancer.

Abstract: Proteinase-activated receptor 2 is a G protein-coupled receptor that regulates vascular tone and inflammatory signalling in the circulatory system. The roles of PAR2 appear complex and sometimes opposing. Studies using PAR2 deficient mice provide a framework to define these effects at the system level. This review examines cardiovascular phenotypes associated with PAR2 deficiency in basal conditions and in disease. PAR2 deficiency produces modest increases in arterial blood pressure and vascular stiffness while preserving endothelial vasodilator function. Cardiac function remains normal in young PAR2 deficient animals but changes with age. Older PAR2 deficient mice develop diastolic dysfunction and cardiac fibrosis. In disease models, PAR2 deficiency promotes fibrosis in cardiac and vascular tissues but reduces vascular inflammation in atherosclerosis. PAR2 deficiency limits plaque progression and promotes stable lesion structure. PAR2 deficiency also reduces myocardial injury and adverse remodelling after ischemia. The effects of PAR2 deficiency on inflammatory signalling vary with context and tissue. These findings show that PAR2 functions depend on physiological and pathological context. Future studies should define cell-specific mechanisms to guide therapeutic targetting of PAR2.

Abstract: Histone-modifying enzymes and histone post-translational modifications (PTMs) play key roles in the organization (euchromatin versus heterochromatin) and function (active versus silenced genes) of chromatin. The abundance and activity of these enzymes, along with their associated histone PTMs, are often altered in cancer cells, leading to deregulated gene expression. The expression of the KMT2A-MLLT3 protein, resulting from a chromosomal translocation in mixed-lineage leukemia (MLL), a subtype of acute myeloid leukemia, augments transcription elongation, promoting the expression of HOXA9 and MEIS1, genes that play critical roles in MLL development. Bortezomib, a proteasome inhibitor, has been effective in treating various cancers. In this study, we compared the impact of bortezomib on histone PTMs in the MLL cell line MOLM-13 and the chronic myeloid leukemic (CML) cell line K562. We report that MOLM-13 had a greater level of histone H2B monoubiquitinated at lysine 120 (H2BK120ub) and histone H3 dimethylated at lysine 79 (H3K79me2) (modifications involved in elongation) and similar levels of histone H2A monoubiquitinated at lysine 119 (H2AK119ub). Bortezomib treatment resulted in significant reductions in H2BK120ub and H2AK119ub levels, the loss of FAcilitates Chromatin Transcription (FACT) chromatin association in both cell lines, and the reduced transcript levels of genes involved in the development of MLL.

Abstract: Metabolic diseases are characterized by a complex interplay between metabolic dysregulation and chronic low-grade inflammation. Fetuin-A (FetA), a liver-derived hepatokine, has emerged as a key mediator linking these processes through its pro-inflammatory and insulin resistance–promoting effects. Accumulating evidence indicates that FetA not only serves as a biomarker but also actively contributes to disease pathogenesis by modulating multiple signaling pathways. In this review, we present an overview of the molecular mechanisms underlying FetA-induced suppression of peroxisome proliferator-activated receptor (PPAR) signaling, a central regulator of metabolic homeostasis. Emerging evidence suggests that FetA may promote Toll-like receptor 4 (TLR4)–mediated inflammation, activate nuclear factor kappa B (NF-κB) signaling, suppress key energy regulators such as Sirtuin 1 (SIRT1) and AMP-activated protein kinase (AMPK), and inhibit PPAR activity through Wnt and extracellular signal-regulated kinase (ERK) pathways. These interconnected mechanisms may contribute to impaired lipid metabolism, increased insulin resistance, and metabolic inflammation. Furthermore, we highlight the role of FetA glycosylation, particularly fucosylation, as a regulatory layer influencing its biological activity. Fucosylated FetA may more effectively activate TLR4 signaling and suppress PPAR activity, suggesting functional heterogeneity among glycoforms. Overall, the FetA–PPAR interaction may represent a key mechanistic link between metabolic inflammation and disease progression.

Abstract: The protist Euglena gracilis has unique and varied metabolic pathways and is an excellent source of dietary protein, essential amino acids, lipids, β-1,3-glucan paramylon, and vitamins. A key concern about using E. gracilis to generate feed and food ingredient products is the possibility that it makes marine toxins. The current investigation therefore used comprehensive analyses of known marine toxin biosynthesis pathways and key genes to evaluate the E. gracilis strain Z for its ability to produce any marine toxins under various fermentation conditions. The major results of this investigation reveals that: 1) E. gracilis does not have the genes essential for the biosynthesis of microcystin, cylindrospermopsin, saxitoxin, brevetoxin, complex toxic polyketides, or domoic acid; 2) E. gracilis does not express an essential gene for the biosynthesis of okadaic acid under heterotrophic fermentation conditions; 3) Phylogenetic analysis using 18S rDNA sequences concluded that E. gracilis is not grouped with any known euglenophycin-producing Euglena species. This study provides a thorough investigation and reviews marine toxins and their genetic mechanisms and metabolic pathways; thus, it can serve as supporting documentation for any feed or food ingredients derived from Euglena gracilis.

Abstract: Bone remodeling in skeletal and dental compartments is governed by shared lineage‑specific programs, yet systemic and local nutrient‑signaling effects are typically reported as isolated, single‑agent findings. This narrative review outlines an integrated mechanistic topology describing how melatonin, strontium, vitamin D3, and vitamin K2 interface with shared osteogenic and inflammatory pathways. By synthesizing evidence across dental pulp stem cells, periodontal ligament stem cells, and osteoblast–osteoclast co‑cultures, this topology positions these agents as parallel mechanistic inputs acting on common signaling nodes. Key regulatory intersections include OPG/RANKL coupling, MAPK/ERK activation, Nrf2‑linked antioxidant defense, and Wnt/β‑catenin‑associated mineralization pathways. This review develops a hypothesis‑generating signaling topology to guide future experimental designs, including dose‑matrix co‑cultures, for testing specific nutrient–pathway interactions. Importantly, this analysis is limited to mechanistic signaling and does not address clinical efficacy or therapeutic supplementation for periodontitis or other dental conditions.

Abstract: The ubiquitin–proteasome system (UPS) has been ‘traditionally’ described as a tightly regulated degradative network driven mainly by the specificity of its ubiquitin-conjugating enzymatic components. The proteasome on the other hand was thought to be a multi-subunit proteolytic complex that recognizes in a non-discriminatory manner ubiquitin-marked target substrates with less than a handful of exceptions. However, emerging evidence reveals that proteasome function is also dynamically regulated by multiple factors such as subunit composition and synthesis, post-translational modifications, and spatial localization which by itself it tightly regulated by the metabolic state of the cell. All these mechanisms add critical regulatory layers to protein homeostasis. This review highlights these newly evolving developments and discusses the pathogenic sequelae of their dysregulation.

Abstract:

Chimeric antigen receptor T-cell (CAR-T) therapy represents a transformative yet high-cost immunotherapeutic strategy that has benefited many patients with hematological malignancies and autoimmune diseases. As of January 2025, only a limited number of CAR-T products have received approval by the FDA, EMA, and the NMPA, primarily targeting CD19 or B-cell maturation antigen (BCMA) expressed on cancer cells. The manufacturing of currently approved CAR-T products predominantly relies on lentiviral vectors (LVVs), largely derived from human immunodeficiency virus type 1 (HIV-1). LVVs are favored due to their high transduction efficiency and their ability to stably integrate transgenes into the genomes of both dividing and non-dividing cells, including post-mitotic mammalian cells, an advantage over gamma retroviral vectors, which exhibit limited capacity to transduce non-dividing cells. This review outlines the fundamental biological principles of HIV-derived LVVs, their structure, functional components, and biotechnological applications. It provides a comparative analysis of different viral vectors, an overview of the CAR construct, and a summary of ex vivo CAR-T manufacturing processes. Additionally, emerging in vivo CAR-T approaches are discussed, with reference to clinically approved LVV-based CAR-T products. Emphasis is placed on microbiological perspectives and environmental biosafety. Finally, recent advances in LVV technology are described, providing insight into the production of next-generation CAR-T therapies employing in vivo gene delivery approaches.

Abstract:

Circadian rhythm disruptions are increasingly recognized in disorders such as obstructive sleep apnea (OSA), yet analysis of 24-hour gene expression patterns remains challenging due to the lack of reliable reference genes for normalization. Even commonly used housekeeping genes may exhibit circadian oscillations, which can confound rhythmic gene expression analyses and hinder biomarker identification. To address this limitation, we evaluated the gene expression stability of 11 commonly used housekeeping genes in blood collected every 6 hours over 24-hour period from 40 adults with varying OSA severity and controls. Stability ranking by analytical tools RefFinder and EndoGeneAnalyzer identified ACTB (β-actin) and RPL13A (ribosomal protein L13a) as the most consistent reference genes, with minimal intra- and inter-individual variability across sampling times and disease groups. Their suitability was assessed by personalized cosinor analysis of core clock genes (BMAL1, PER2, CRY1), demonstrating that appropriate normalization enables detection of circadian oscillations in clinical samples. Using the optimal normalization, CosinorPy analysis of the core clock genes revealed significant circadian oscillations of at least one clock gene in the studied participants. These findings establish ACTB and RPL13A as robust reference genes for blood-based circadian studies of OSA and provide an important methodological framework for future circadian biomarker research.