Phosphorylation Process Research Articles

Selection increases mitonuclear DNA discordance but reconciles incompatibility in African cattle.

Mitochondrial function relies on the coordinated interactions between genes in the mitochondrial (mtDNA) and nuclear genomes. Imperfect interactions following mitonuclear incompatibility may lead to reduced fitness. MtDNA introgressions across species and populations are common and well documented. Various strategies may be expected to reconcile mitonuclear incompatibility in hybrids or admixed individuals. African admixed cattle (Bos taurus × B. indicus) show sex-biased admixture, with taurine (B. taurus) mtDNA and a nuclear genome predominantly of humped zebu (B. indicus). Here, we leveraged local ancestry inference approaches to identify the ancestry and distribution patterns of nuclear functional genes associated with the mitochondrial oxidative phosphorylation process in the genomes of African admixed cattle. We show that most of the nuclear genes involved in mitonuclear interactions are under selection and of humped zebu ancestry. Variation in mtDNA copy number (mtDNA-CN) may have contributed to the recovery of optimal mitochondrial function following admixture with the regulation of gene expression, alleviating or nullifying mitochondrial dysfunction. Interestingly, some nuclear mitochondrial genes with enrichment in taurine ancestry may have originated from ancient African aurochs (B. primigenius africanus) introgression. They may have contributed to the local adaptation of African cattle to pathogen burdens. Our study provides further support and new evidence showing that the successful settlement of cattle across the continent was a complex mechanism involving adaptive introgression, mtDNA-CN variation, regulation of gene expression, and selection of ancestral mitochondria-related genes.

Near Infrared-Mediated Intracellular NADH Delivery Strengthens Mitochondrial Function and Stability in Muscle Dysfunction Model.

Mitochondrial transfer emerges as a promising therapy for the restoration of mitochondrial function in damaged cells, mainly due to its limited immunogenicity. However, isolated mitochondria rapidly lose function because they produce little energy outside cells. Therefore, this study investigates whether near infrared (NIR)-mediated nicotinamide adenine dinucleotide (NADH) pre-treatment enhances mitochondrial function and stability in mitochondria-donor cells prior to transplantation. Clinical applications of NADH, an essential electron donor in the oxidative phosphorylation process, are restricted due to the limited cellular uptake of NADH. To address this, a photo-mediated method optimizes direct NADH delivery into cells and increases NADH absorption. L6 cells treated with NADH and irradiated with NIR enhanced NADH uptake, significantly improving mitochondrial energy production and function. Importantly, the improved functional characteristics of the mitochondria are maintained even after isolation from cells. Primed mitochondria, i.e., those enhanced by NIR-mediated NADH uptake (P-MT), are encapsulated in fusogenic liposomes and delivered into muscle cells with mitochondrial dysfunction. Compared to conventional mitochondria, P-MT mitochondria promote greater mitochondrial recovery and muscle regeneration. These findings suggest that NIR-mediated NADH delivery is an effective strategy for improving mitochondrial function, and has the potential to lead to novel treatments for mitochondrial disorders and muscle degeneration.

Joint exploration of network pharmacology and metabolomics on the effects of traditional Chinese medicine compounds in weaned yaks.

Chinese herbal medicines are relatively inexpensive and have fewer side effects, making them an effective option for improving health and treating diseases. As a result, they have gained more attention in recent years. The weaning period is a critical stage in the life of yaks, often inducing stress in calves. Weaning stress, along with dietary changes, can lead to a decline in physical fitness and immune function, making yaks more susceptible to diarrhea and resulting in high mortality rates during this period. Therefore, our study aimed to address this issue by incorporating traditional Chinese medicine (TCM) formulas into the diet of yaks during the weaning period. Following a dialectical analysis, three TCM formulas, mainly composed of Paeonia lactiflora, Coptis chinensis, and Dandelion, were identified for their anti-inflammatory, antioxidant, and immune enhancing potentials. We explored the possible molecular mechanisms of these TCM formulas using network pharmacology analysis and investigated their effects on the physiology of yaks through metabolomics. Network pharmacology analysis revealed several key target proteins in the protein-protein interaction (PPI) network between three formulas and immune-related genes, including PIK3R1, PIK3CA, JAK2, PTK2, and PYPN11. The key target proteins in the PPI network associated with metabolism-related genes included ENPP1, CYP1A1, PTGS1, members of the CYP1 family, and EPHX2. GO analysis of co-targets revealed highly enriched pathways such as protein phosphorylation, plasma membrane, and one-carbon metabolic processes. Metabolomics revealed significant changes in the abundance of metabolites including dimethyl sulfoxide, tyrphostin A25, and thromboxane A2 in the intestines of weaned yaks supplemented with these Chinese herbal compounds. Significant changes were also observed in pathways such as vitamin A metabolism, chloroalkane, and chloroalkene degradation. Based on these findings, it can be inferred that TCM formulas improve the physical fitness of weaned yaks by enhancing antioxidant capacity, boosting immunity, and reducing intestinal inflammation. This study preliminarily elucidates the pharmacological mechanisms by which TCM formulas prevent diarrhea and improve physical fitness in weaned yaks through metabolomics and network pharmacology, paving the way for further evaluation of the effectiveness of these three formulas.

C/EBPβ in Alzheimer's disease: An integrative regulator of pathological mechanisms.

Alzheimer's disease (AD) stands as one of the most prevalent neurodegenerative disorders, characterized by a progressive decline in cognitive function, neuroinflammation, amyloid-beta (Aβ) plaques, and neurofibrillary tangles (NFTs). With the global aging population, the incidence of AD continues to rise, yet current therapeutic strategies remain limited in their ability to significantly alleviate cognitive impairments. Therefore, a deeper understanding of the molecular mechanisms underlying AD is imperative for the development of more effective treatments. In recent years, the transcription factor C/EBPβ has emerged as a pivotal regulator in several pathological processes of AD, including neuroinflammation, lipid metabolism, Aβ processing, and tau phosphorylation. Through intricate post-translational modifications, C/EBPβ modulates these processes and may influence the progression of AD on multiple fronts. This review systematically explores the multifaceted roles of C/EBPβ in the pathogenesis of AD, delving into its crucial involvement in neuroinflammation, Aβ production, tau pathology, and lipid metabolism dysregulation. Furthermore, we critically assess therapeutic strategies targeting C/EBPβ, examining the challenges and opportunities in regulating this factor. By synthesizing the latest research findings, we offer a more comprehensive understanding of the role of C/EBPβ in AD and discuss its potential as a therapeutic intervention target.

Liquiritin Ameliorates Palmitic Acid‐Induced Hepatic Steatosis in Mouse Primary Hepatocytes by Suppressing the VEGFA Signaling Pathway

Nonalcoholic fatty liver disease (NAFLD) has become a global pandemic, imposing a significant socioeconomic burden. Hepatic steatosis is a key pathological event in NAFLD. However, there is still lack of effective drugs for NAFLD treatment. Liquiritin (LIQ), derived from licorice root, exhibits antioxidative and anti‐inflammatory properties, making it valuable in managing various conditions such as dermatological disorders, respiratory ailments, and gastritis. Here, we investigated the impact and underlying mechanisms of LIQ on hepatic steatosis in mouse primary hepatocytes (MPHs). Our study found that LIQ significantly decreased palmitic acid (PA)‐induced lipid accumulation in MPHs. Additionally, LIQ remarkably increased the mRNA expression of Cpt1a, PPARα, Ehhadh, Cyp4a10, and Acox1, suggesting that LIQ achieves its lipid‐lowering effect by regulating mitochondria‐mediated lipid β‐oxidation. Furthermore, the regulatory effect of LIQ on the mitochondrial oxidative phosphorylation (OXPHOS) process was confirmed by Seahorse analysis. Mechanistically, bioinformatics analysis predicted VEGFA as a crucial target mediating the beneficial effects of LIQ against PA‐induced lipid accumulation in MPHs. Notably, treatment with purified VEGFA protein partially counteracted the lipid‐lowering properties of LIQ. Additionally, our data showed that LIQ promoted VEGFA degradation through the ubiquitin–proteasome pathway. Therefore, our findings not only confirm the lipid‐lowering effects of LIQ in MPHs but also identify VEGFA as its potential target. These results highlight the therapeutic promise of LIQ in managing NAFLD and introduce VEGFA as a novel target for treating hepatic steatosis.

Revisiting phosphoregulation of Cdc25C during M-phase induction.

Cdc25C undergoes a sudden and substantial gel mobility shift at M-phase onset, correlating with abrupt activation of both Cdc25C and Cdk1 activities. A positive feedback loop between Cdk1 and Cdc25C has been used to explain this hallmark phenomenon. Here, we demonstrate that the M-phase supershift and robust activation of Cdc25C are due to the site-comprehensive phosphorylation of its long intrinsically disordered regulatory domain without requiring Cdk1 or other major mitotic kinase activities. The phosphorylation process involves substrate-mediated assembly of phosphorylation machinery that catalyzes multisite phosphorylation continuously without substrate dissociation. In contrast to the site-comprehensive phosphorylation of Cdc25C occurring at M-phase onset, the site-specific phosphorylation of Cdc25C by Cdk1 or other major mitotic kinases generates slight gel mobility shifts and modest activation of Cdc25C prior to M-phase onset. These findings suggest a two-stage framework consisting of site-specific phosphorylation followed by site-comprehensive phosphorylation for Cdc25C regulation during M-phase induction.

The protection of nicotinamide riboside against diabetes mellitus-induced bone loss via OXPHOS.

Diabetes mellitus is a global disease that results in various complications, including diabetic osteoporosis. Prior studies have indicated a correlation between low levels of nicotinamide adenine dinucleotide (NAD+) and diabetes-related complications. Nicotinamide riboside (NR), a widely utilized precursor vitamin of NAD+, has been demonstrated to enhance age-related osteoporosis through the Sirt1/FOXO/β-catenin pathway in osteoblast progenitors. However, the impact of NR on bone health in diabetes mellitus remains unclear. In this study, we assessed the potential effects of NR on bone in diabetic mice. NR was administered to high-fat diet (HFD)/streptozotocin (STZ)-induced type 2 diabetic mice (T2DM), and various parameters, including metabolic indicators, bone quality, bone metabolic markers, and RNA sequences, were measured. Our findings confirmed that HFD/STZ-induced T2DM impaired bone microstructures, resulting in bone loss. NR effectively ameliorated insulin resistance, improved bone microarchitecture, and bone quality, reduced bone resorption, enhanced the Forkhead box O (FOXO) signaling pathway, mitigated the nuclear factor kappa B (NF-kB) signaling pathway, and ameliorated the disorder of the oxidative phosphorylation process (OXPHOS) in diabetic mice. In conclusion, NR demonstrated the capacity to alleviate T2DM-induced bone loss through the modulation of OXPHOS in type 2 diabetic mice. Our results underscore the potential of NR as a therapeutic target for addressing T2DM-related bone metabolism and associated diseases. Further cell-based studies under diabetic conditions, such as in vitro cultures of key cell types (e.g., osteoblasts and osteoclasts), are necessary to validate these findings.

Phosphorus-oxygen bonded CoZn catalysts for efficiently boosting oxygen evolution activity of BiVO4 photoanodes

Designing and constructing a robust interaction at photoanode/cocatalyst interface to achieve high activity is a pressing necessity for the advancement of photoelectrochemical water splitting technology. Herein, the modification of BiVO4 photoanodes with MOF-derived CoZnPOx cocatalysts was successfully achieved through the application of precipitation and low-temperature phosphorylation processes. A high photocurrent density of 3.72 mA/cm2 at 1.23 V (vs. reversible hydrogen electrode (RHE)) was obtained, accompanied by a negative shift in the onset potential (approximately 174 mV). The incorporation of P–O bonds has been demonstrated to result in a redistribution of electron density and the formation of strong electronic interactions between CoZnPOx and BiVO4 photoanode. These changes have been shown to significantly improve hole mobility, facilitate charge separation and optimize the kinetic processes, thereby enhancing the overall performance of the water oxidation reaction.

Protein serine/threonine phosphatases in tumor microenvironment: a vital player and a promising therapeutic target.

The tumor microenvironment (TME) is involved in cancer initiation and progression. With advances in the TME field, numerous therapeutic approaches, such as antiangiogenic treatment and immune checkpoint inhibitors, have been inspired and developed. Nevertheless, the sophisticated regulatory effects on the biological balance of the TME remain unclear. Decoding the pathological features of the TME is urgently needed to understand the tumor ecosystem and develop novel antitumor treatments. Protein serine/threonine phosphatases (PSPs) are responsible for inverse protein phosphorylation processes. Aberrant expression and dysfunction of PSPs disturb cellular homeostasis, reprogram metabolic processes and reshape the immune landscape, thereby contributing to cancer progression. Some therapeutic implications, such as the use of PSPs as targets, have drawn the attention of researchers and clinicians. To date, the effects of PSP inhibitors are less satisfactory in real-world practice. With breakthroughs in sequencing technologies, scientists can decipher TME investigations via multiomics and higher resolution. These benefits provide an opportunity to explore the TME in a more comprehensive manner and inspire more findings concerning PSPs in the TME. The current review starts by introducing the canonical knowledge of PSPs, including their members, structures and posttranslational modifications for activities. We then summarize the functions of PSPs in regulating cellular homeostasis. In particular, we specified the up-to-date roles of PSPs in modulating the immune microenvironment, adopting hypoxia, reprogramming metabolic processes, and responding to extracellular matrix remodeling. Finally, we introduce preclinical PSP inhibitors with translational value and conclude with clinical trials of PSP inhibitors for cancer treatment.

Chronic unpredictable stress induces anxiety-like behavior and oxidative stress, leading to diminished ovarian reserve

Chronic stress can adversely affect the female reproductive endocrine system, potentially leading to disorders and impairments in ovarian function. However, current research lacks comprehensive understanding regarding the biochemical characteristics and underlying mechanisms of ovarian damage induced by chronic stress. We established a stable chronic unpredictable stress (CUS)-induced diminished ovarian reserve (DOR) animal model. Our findings demonstrated that prolonged CUS treatment over eight weeks resulted in increased atresia follicles in female mice. This atresia was accompanied by decreased AMH and increased FSH levels. Furthermore, we observed elevated levels of corticosterone both in the peripheral blood and within the ovary. Additionally, we detected abnormalities in ATP metabolism within the ovarian tissue. CUS exposure led to oxidative stress in the ovaries, fostering a microenvironment characterized by oxidative damage to mouse ovarian granulosa cells (mGCs) and heightened levels of reactive oxygen species. Furthermore, CUS prompted mGCs to undergo apoptosis via the mitochondrial pathway. These findings indicate a direct association between the fundamental physiological alterations leading to DOR and the oxidative phosphorylation processes within mGCs. The diminished ATP production by mGCs, triggered by CUS, emerges as a pivotal indicator of CUS-induced DOR. Our study establishes an animal model to investigate the impact of chronic stress on ovarian reserve function and sheds light on potential mechanisms underlying this phenomenon.

Enhanced Water Splitting Electrocatalysis with Heterointerfacial Cobalt Phosphide In Situ Supported on a Cellulose-Derived Carbon Aerogel.

The active site density, intrinsic activity, and supporting substrate of cobalt phosphide catalysts are vital to their performance in alkaline water electrolysis. In this work, a CoP/Co2P loaded on cellulose nanofiber-derived carbon aerogels (CP/CCAs) bifunctional electrocatalyst with a three-dimensional network and heterostructure is illustrated through sequential facile hydrothermal, freeze-drying, and phosphorylation processes. The three-dimensional network of carbon aerogels derived from cellulose nanofibers reveals a specific surface area of 183.41 m2/g, greatly enriching the active sites and facilitating the electron and mass transportation. Besides, interactions between CoP/Co2P with a heterogeneous structure and carbon aerogels modulate the electronic structure to enhance the intrinsic activity of the catalysts. Benefiting from these advantages, CP/CCAs-2 demonstrates a superior oxygen evolution reaction activity (η10 mA cm-2 = 277 mV) over the benchmark RuO2 and a moderate hydrogen evolution reaction performance (η10 mA cm-2 = 63 mV). Density functional theory calculations further reveal that the coupling of CoP/Co2P and cellulose-derived carbon aerogels promotes the water adsorption and activates the H-O bond. As a result, the alkaline electrolyzer assembled with CP/CCAs-2 both as a cathode and an anode shows a low cell voltage of 1.59 V at a current density of 10 mA cm-2 and good stability. This work provides a strategy for phosphide electrocatalysts composited with green carbon carriers for overall water splitting.

Role of Bioenergy Effects of Cystamine in Realising Potentiation of Radioprotective Properties of the Radioprotector in its Repeated Administration

Early phenomenon on the potentiation of radioprotective properties of aminothiols (cystamine) during repeated application in the first 7 h of drug’s after-effect by increasing their activity by 2 times was discovered (B.I. Davydov, 1971, M.V. Vasin, V.V. Antipov, 1972). A hypothesis has been proposed to explain this phenomenon. The mechanism of radioprotective properties cystamine is associated with partial neutralization of oxygen effect with the development of reductive stress in the cell, which does not end after the cessation of protective effect of radioprotector. In the body, negative feedback mechanisms are implemented at the cell, which prevent full implementation of naked consequences of reductive stress. In it, the transcription factor HIF-1 plays a key role, which enhances the production of ATP due to glycolysis and, thereby, reduces the burden on oxidative phosphorylation processes under conditions of acute hypoxia with repeated use of cystamine over the first 7 h after the cessation of its radioprotective activity. There is an increase in metabolic shifts in the body with the development of deep hypothermia in animals up to 29°C rectal temperature which can lead to depletion of these restrictive mechanisms and, thereby allowing aminothiol to fully exhibit its protective properties without excluding the increase in its toxicity. In addition, this contributes to a more complete implementation of post-radiation repair of DNA. These processes contribute to a more complete realization of the affected repair of DNA breaks by providing more time for it in conditions of longer hypothermia and mitotic blockade under the action of cystamine.

Role of PIM Kinase Inhibitor in the Treatment of Alzheimer's Disease.

Alzheimer's disease (AD), a neurodegenerative disorder, is the most prevalent form of senile dementia, causing progressive deterioration of cognition, behavior, and rational skills. Neuropathologically, AD is characterized by two hallmark proteinaceous aggregates: amyloid beta (Aβ) plaques and neurofibrillary tangles (NFTs) formed of hyperphosphorylated tau. A significant study has been done to understand how Aβ and/or tau accumulation can alter signaling pathways that affect neuronal function. A conserved protein kinase known as the mammalian target of rapamycin (mTOR) is essential for maintaining the proper balance between protein synthesis and degradation. Overwhelming evidence shows mTOR signaling's primary role in age-dependent cognitive decline and the pathogenesis of AD. Postmortem human AD brains consistently show an upregulation of mTOR signaling. Confocal microscopy findings demonstrated a direct connection between mTOR and intraneuronal Aβ42 through molecular processes of PRAS40 phosphorylation. By attaching to the mTORC1 complex, PRAS40 inhibits the activity of mTOR. Furthermore, inhibiting PRAS40 phosphorylation can stop the Aβ-mediated increase in mTOR activity, indicating that the accumulation of Aβ may aid in PRAS40 phosphorylation. Physiologically, PRAS40 is phosphorylated by PIM1 which is a serine/threonine kinase of proto-oncogene PIM kinase family. Pharmacological inhibition of PIM1 activity prevents the Aβ-induced mTOR hyperactivity in vivo by blocking PRAS40 phosphorylation and restores cognitive impairments by enhancing proteasome function. Recently identified small-molecule PIM1 inhibitors have been developed as potential therapeutic to reduce AD-neuropathology. This comprehensive study aims to address the activity of PIM1 inhibitor that has been tested for the treatment of AD, in addition to the pharmacological and structural aspects of PIM1.

Mitochondrial Dysfunction and Metabolic Indicators in Patients with Drug-Naive First-Episode Schizophrenia: A Case-Control Study.

This paper aims to explore the expression characteristics of mitochondrial function-related genes in patients with first-episode schizophrenia (SCZ)and the correlation between differentially expressed genes and clinical metabolic indicators. Twenty patients with first-episode SCZ who had not taken antipsychotic drugs (patient group) and twenty healthy controls (control group) were included. Quantitative real-time PCR technology was used to detect the expression levels of genes related to mitochondrial quality control and oxidative phosphorylation in peripheral blood leukocytes, and metabolic indicators such as blood biochemistry and blood glucose were collected. The gene expression levels of key genes related to mitochondrial function, PGC-1a, PARK2, and LC3B, in the patient group were significantly lower than those in the control group (P < 0.05). Correlation analysis showed that the expression level of PGC-1a gene in the patient group was negatively correlated with very low-density lipoprotein levels (r =-0.451), and the expression level of PARK2 gene in the patient group was negatively correlated with uric acid levels (r =-0.447). The expression levels of multiple key genes in the mitochondrial quality control and oxidative phosphorylation processes in patients with first-episode SCZ display a downward trend. The differentially expressed genes are correlated with the metabolic abnormalities of the patients, suggesting that mitochondrial dysfunction may be related to the high incidence of metabolic diseases in patients with SCZ.

Structural features, physiological functions and digestive properties of phosphorylated corn starch: A comparative study of four phosphorylating agents and two preparation methods.

Phosphorylation is an important modification to modulate functional and digestive properties of starches. We systematically investigated starch phosphorylation process parameters by using two different preparation methods (slurry and semi-dry conditions) and four commonly used phosphorylating agents, namely sodium tripolyphosphate (STPP), sodium trimetaphosphate (STMP), STMP/STPP (99: 1), and sodium phytate (SP). The effects of phosphorylation on physicochemical characteristics, techno-functionalities, digestive properties and structural features of corn starch were analyzed. Phosphorylation with the semi-dry method resulted in higher phosphorus content, degree of double helix, and degree of starch aggregation and lower amylose content and relative crystallinity than with the slurry method. Phosphorylation using semi-dry conditions, irrespective of the used phosphorylating agent, furthermore decreased the gelatinization temperature, enthalpy, the temperature corresponding to the maximum starch mass loss rate and estimated glycemic index of corn starch, and increased solubility, swelling power, peak viscosity, transmittance, and resistant starch content. Of the phosphorylating agents, independent of the used preparation method, STMP and STMP/STPP resulted in the highest degrees of starch phosphorylation and therefore modulated the physiochemical, functional and digestive properties of corn starch more than STPP and SP. The findings of this systematic comparison provide important information to tailor phosphorylated corn starches to meet specific food requirements.

Lithocholic Acid's Ionic Compounds as Promising Antitumor Agents: Synthesis and Evaluation of the Production of Reactive Oxygen Species (ROS) in Mitochondria.

The development of a methodology for the synthesis of new compounds with antitumor activity represents a significant and priority task within the field of medicinal chemistry. As a continuation of our research group's earlier studies on the antitumor activity of ionic derivatives of natural compounds, we have synthesized a series of previously undescribed pyrazole ionic compounds through a series of transformations of lithocholic acid methyl ester. To investigate the biological activity of the newly synthesized lithocholic acid derivatives, a series of modern flow cytometry techniques were employed to assess their cytotoxic activity, effects on the cell cycle, and induction of apoptosis. This included the analysis of alterations in the mitochondrial potential, accumulation of ROS ions in mitochondria, and loss of cytochrome c. These compounds demonstrate promising antitumor activity through their effects on mitochondrial oxidation and phosphorylation processes. These compounds, which we have designated as "soft dissociators", exhibit enhanced biopharmacological properties relative to the original lithocholic acid molecule.

Fe, P co-doped hybrid electrocatalyst for synergistic enhancement of electrocatalytic hydrogen evolution reaction durability performance

Defect engineering has been widely applied to improve the hydrogen evolution reaction (HER) performance of MoS2. In this work, a co-doped electrocatalyst on carbon fiber paper (CFP) for HER was prepared by coupling with simple hydrothermal and gas-phase phosphorylation process to improve the durability of the catalyst while enhancing the electrocatalytic performance (Fe-P-MoS2/CFP). The results showed that the overpotential at a current density of 10 mA cm−2 (η10) of Fe-P-MoS2/CFP was only 130 mV, which was much lower than those of other undoped and single-metal atom doping electrocatalysts. Electronic interactions between Fe, P and MoS2 reduced the local electron densities and changed the electronic structure of Mo and S, leading to the generation of additional p orbitals in the S site, thus optimizing the adsorption–desorption energy of hydrogen in the S site. In addition, compared to Fe-MoS2/CFP, Fe-P-MoS2/CFP showed better electrocatalytic durability in acidic conditions. The co-doping technique proposed within this study stands to offer a novel approach for amplifying the catalytic activity and stability of electrocatalysts.

Effect of cold atmospheric plasma (CAP) on the energy metabolism in HeLa cells through miRNA regulation

Abstract Cold atmospheric plasma (CAP) is an emerging tool for tumor treatment because it can inhibit cancer cell proliferation primarily through oxidative stress due to CAP-induced reactive oxygen species (ROS). Among various ROS targeting molecules in cancer cells, microRNAs (miRNAs) are one kind of important targets that can be stimulated by ROS, and many studies have shown that miRNAs are involved in the metabolism regulation of cancer cells. In this study, we applied helium-CAP (He-CAP) to HeLa cells, and observed that the ROS induced by He-CAP could modulate the miRNAs related to energy metabolism, leading to the changes of proliferation, glycolysis, tricarboxylic acid (TCA) cycling and oxidative phosphorylation (OXPHOS) in the HeLa cells, and affected the related hypoxia-inducible factor 1, p53, phosphoinositide 3-kinase signaling pathways. In addition, the analysis of miRNAs in the metabolic network revealed that the expressions of the miRNAs responsible for the promotion of energy metabolism increased, and correspondingly, the involved mRNA and protein expression decreased. As such, this study has not only demonstrated that CAP treatment could significantly change the miRNAs expression of cancer cells, but also provided a more in-depth understanding of the CAP effects on glycolysis, TCA cycling and OXPHOS processes in the cells through the comprehensive analysis of the miRNAs regulation.

GdX inhibits the occurrence and progression of breast cancer by negatively modulating the activity of STAT3

ABSTRACT Aim To elucidate the biological functionality and regulatory mechanisms of GdX in breast cancer (BC). Methods The examination of GdX expression in human BC tissues and cell lines was conducted through immunohistochemical (IHC) and Western blot. Cell proliferation capacity was assessed via the CCK-8 and colony formation assay, while cell migration was determined through the wound healing assay. The expression levels of BCL-XL, Cyclin D1, and C-myc gene were quantified using RT-qPCR and Western blot. In vivo tumor growth was evaluated in nude mice xenografted with MDA-MB-231 cells overexpressing GdX, and a mouse model with GdX-deficient BC was established to observe the impact of GdX on BC formation and metastasis. Dual-luciferase reporter assay and immunofluorescence were employed to confirm the interaction between GdX and STAT3. Western blot was employed to validate the influence of GdX overexpression on the phosphorylation process of STAT3. Results GdX exhibited low expression in the cancer tissues of BC patients and cell lines. MDA-MB-231 and MCF-7 cells overexpressing GdX displayed a notable reduction in proliferation and diminished migratory capabilities, accompanied by downregulated mRNA and protein expression of BCL-XL, Cyclin D1, and C-myc. In the xenograft mouse model, heightened GdX expression correlated with a decelerated in vivo tumor growth. Furthermore, in mice deteleing GdX, both the quantity and weight of tumors increased, along with evident pulmonary metastasis. Mechanistically, STAT3 emerged as a downstream target gene of GdX. Conclusions GdX exerts its inhibitory effects on the initiation and progression of BC by negatively modulating the phosphorylation of STAT3.

Protein Engineering of Nicotinamide Riboside Kinase Based on a Combinatorial Semirational Design Strategy for Efficient Biocatalytic Synthesis of Nicotinamide Mononucleotides.

Industrial biosynthesis of β-nicotinamide mononucleotide (β-NMN) lacks a highly active nicotinamide riboside kinase for the phosphorylation process. Cumbersome preprocessing steps and excessive ATP addition contribute to its increased cost. To tackle these challenges, a docking combination simulation (DCS) semirational mutagenesis strategy was designed in this study, combining various modification strategies to obtain a mutant NRK-TRA with 2.9-fold higher enzyme activity. Molecular dynamics simulations and structural analysis demonstrate the enhancement of its structural stability. High-density fermentation was achieved through a 5 L fermentation tank, with a titer reaching 208.3 U/mL, the highest in the current report. An ATP-cycling whole-cell catalytic system was employed and optimized by introducing a polyphosphate kinase 2 (PPK2) recombinant strain, and 15.16 g/L β-NMN was obtained through a series of batch transformation experiments. This study provides a new strategy for the efficient screening of highly active enzyme variants and offers a green and promising biotransformation system for NMN production.