acetyl-CoA Research Articles

A high-resolution bovine mitochondrial co-expression network.

The mitochondrion is a sophisticated, versatile, and dynamic organelle whose function is incompletely understood. Intending to provide a framework for mitochondrial visualisation and interpretation of genome-wide molecular data, we reverse-engineered a co-expression network whose final structure represented mRNA encoding more than half of the entire mitochondrial proteome. We drew upon 723 RNA-seq data sets representing 91 tissues and cell types from 441 individual cattle. A mitochondrial landscape was formed comprising a main network and many smaller sub-networks. One of the discrete sub-networks contains all 13 mRNA (e.g. MT-ND1, MT -CYTB, MT -COX2, MT -ATP8) plus 15/22 tRNA (e.g. MT-TT) encoded by the mt-genome itself, indicating some independent regulation from the nuclear genome with whom it must cooperate. Intriguingly, this mtDNA sub-network also contains a single nuclear-encoded gene, that of PDHA1. PDHA1 encodes a subunit of the pyruvate dehydrogenase complex that governs the conversion of pyruvate to Acetyl CoA. This enzyme is extremely influential, representing the fundamental cellular connection between the ancient, conserved pathway of glycolysis that occurs exclusively in the cytoplasm, and the TCA cycle that occurs within the mitochondrial matrix. To demonstrate the downstream utility of our approach, we overlaid Longissimus dorsi muscle transcriptome data from differentially feed efficient Charolais and Holstein Friesian cattle. This approach highlighted expression patterns sensitive to both breed and diet in a complex manner. An analytic advantage of this approach is that relatively subtle (<2-fold) but coordinated changes that may be overlooked by conventional gene-by-gene significance testing become readily apparent. Finally, intending to understand the transcriptional regulation of mitochondrial function more thoroughly, we engineered a network built with transcription factors in addition to those mRNA encoding mitochondrial proteins. Here, a set of influential nuclear hormone receptors (e.g. PPARA) are enriched among the most highly and/or well-connected TF.

Preliminary Investigation on Resistance of Beckmannia syzigachne to Clodinafop-Propargyl and Mesosulfuron-Methyl from China

Beckmannia syzigachne is one of the most competitive weeds in winter wheat fields in China. In this study, 120 suspected resistant populations of Beckmannia syzigachne were collected from the Anhui, Hubei, Jiangsu, and Shandong Provinces from 2017 to 2019. In total, 110 populations exhibited different levels of resistance to clodinafop-propargyl, 114 populations expressed different levels of resistance to mesosulfuron-methyl, and 105 populations were resistant to both herbicides at different levels. The resistant weeds were mainly distributed in Anhui and Jiangsu Provinces. The detection results of acetyl coA carboxylase (ACCase) and acetolactate synthase (ALS) genes in the resistant populations indicated that ACCase gene mutations occurred in 97 out of 110 populations resistant to clodinafop-propargyl and ALS gene mutations occurred in 25 out of 114 populations resistant to mesosulfuron-methyl. There were several mutation types, including Ile-1781-Leu, Trp-2027-Cys, Ile-2041-Asn, Ile-2041-Val, Asp-2078-Gly, and Gly-2096-Ala in the ACCase sequence and Pro-197-Ser, Pro-197-Thr, Pro-197-His, Pro-197-Leu, Asp-376-Glu, and Trp-574-Leu in the ALS sequence. Among these mutation types, Pro-197-His, Asp-376-Glu, and Trp-574-Leu in the ALS sequence were the first identified in Beckmannia syzigachne.

DNA methylation, histone acetylation in the regulation of memory and its modulation during aging.

Memory formation is associated with constant modifications of neuronal networks and synaptic plasticity gene expression in response to different environmental stimuli and experiences. Dysregulation of synaptic plasticity gene expression affects memory during aging and neurodegenerative diseases. Covalent modifications such as methylation on DNA and acetylation on histones regulate the transcription of synaptic plasticity genes. Changes in these epigenetic marks correlated with alteration of synaptic plasticity gene expression and memory formation during aging. These epigenetic modifications, in turn, are regulated by physiology and metabolism. Steroid hormone estrogen and metabolites such as S-adenosyl methionine and acetyl CoA directly impact DNA and histones' methylation and acetylation levels. Thus, the decline of estrogen levels or imbalance of these metabolites affects gene expression and underlying brain functions. In the present review, we discussed the importance of DNA methylation and histone acetylation on chromatin modifications, regulation of synaptic plasticity gene expression and memory consolidation, and modulation of these epigenetic marks by epigenetic modifiers such as phytochemicals and vitamins. Further, understanding the molecular mechanisms that modulate these epigenetic modifications will help develop recovery approaches.

Discovery and mechanism of a highly selective, antifungal acetyl CoA synthetase inhibitor.

Acetyl CoA synthetases (ACS) have emerged as drug targets for the treatment of cancer, metabolic diseases as well as fungal and parasitic infections. Although a variety of small molecule ACS inhibitors have been discovered, the systematic optimization of these molecules has been slowed by a lack of structural information regarding their mechanism of inhibition. Through a chemical genetic-based, synthetic lethal screen of the human fungal pathogen Cryptococcus neoformans, we identified an isoxazole-based ACS inhibitor with antifungal activity and exquisite selectivity for the C. neoformans Acs1 relative to human ACSS2 as well as other fungal ACSs. Xray crystallographic characterization of the isoxazole-CnAcs1 complex revealed that the isoxazole functions as an acetyl CoA mimic and occupies both the acetyl- and CoA-binding sites of CnAcs1. Consistent with this novel mode of inhibition, the isoxazoles display uncompetitive inhibition kinetics that are similar to antimalarial ACS inhibitors also proposed to target the CoA binding site. Consequently, these data provide structural and mechanistic insights into the remarkable selectivity of Acetyl CoA pocket-targeting ACS inhibitors. In addition, these data provide strong proof-of-principle that targeting fungal and parasitic ACSs for the development of novel anti-infectives can be achieved with high selectivity and, thereby, low host toxicity.

Olanzapine exposure disordered lipid metabolism, gut microbiota and behavior in zebrafish (Danio rerio).

Olanzapine (OLZ) is widely used in the treatment of schizophrenia, and its metabolic side effects have garnered significant attention in recent years. Despite this, the specific side effects of OLZ and the underlying mechanisms remain inadequately understood. To address this gap, zebrafish (Danio rerio) were exposed to OLZ at concentrations of 35.5, 177.5, and 355.5μg/L. The results indicated that exposure to OLZ significantly increased body weight, total cholesterol (TC), low-density lipoprotein (LDL), and triglycerides (TG). Histological analysis revealed notable lipid accumulation in the liver. Furthermore, lipid synthesis genes, including sterol regulatory element binding protein (srebp), acetyl CoA carboxylase (acc), and fatty acid synthesis gene (fas), were up-regulated. In contrast, genes related to lipid decomposition, such as lipoprotein lipase (lpl), hormone-sensitive triglyceride lipase (hsl), and carnitine palmitoyltransferase 1b (cpt1b), were down-regulated. Subsequent analysis of zebrafish behavior showed reduced motor activity, sociability, and anxiety-like behavior in OLZ-exposed zebrafish, consistent with the results of neurotransmitter related gene expression. Following OLZ treatment, the expression of tryptophan hydroxylase (tph), tyrosine hydroxylase (th), dopamine transporter (dat), glutaminase (glsa), and glutamic acid decarboxylase 1b (gad1b) was upregulated. Additionally, the diversity of intestinal flora decreased after OLZ exposure, and the structure of the intestinal microbiota changed significantly compared to the control group. At the genus level, the abundance of Plesiomonas was upregulated, while the abundances of Bacillus and Cetobacterium were downregulated in the OLZ-exposed group. Furthermore, the results of the correlation analysis indicated that lipid metabolism and behavioral changes were closely associated with the microbiota. This study clarified the side effects of OLZ, and also provided a basis for the reasonable discharge concentration of OLZ in water and clinical drug use.

Negative regulation of SREBP-1/FAS signaling molecules activates the RIG-1/TBK1-mediated IFN-I pathway to inhibit BVDV replication.

For many viruses, controlling the process of infection is largely dependent on the enzymes of the fatty acid synthesis (FAS) pathway. An appealing therapeutic target in antiviral research is fatty acid synthetase (FASN), a crucial enzyme in the FAS pathway. Bovine viral diarrhea, caused by the Bovine viral diarrhea virus (BVDV), is a significant viral infectious disease posing a substantial threat to global animal husbandry. Our study revealed that BVDV infection not only upregulates the expression of FAS-related enzymes in BT cells and the blood, liver, and spleen of mice but also markedly enhances the accumulation of lipid droplets, free fatty acids, and triglycerides. The FAS pathway plays a pivotal role throughout the entire BVDV replication cycle. Additionally, administration of the FASN inhibitor C75 and Acetyl CoA carboxylase-1 (ACC-1) inhibitor TOFA significantly reduced the viral content in both serum and organs of BVDV-infected mice, exhibiting inhibitory effects across diverse viral strains. Intriguingly, We found that RIG-1/TBK1-mediated IFN-I signaling inhibits SREBP-1/FAS and reduces BVDV replication. Conversely, targeting a few essential enzymes of SREBP-1/FAS also activates IFN-I signaling. More importantly, FASN inhibitor led to heightened expression of ISGs in mouse spleens by activating the RIG-1/TBK-1 pathway. These findings highlight that FASN inhibitors inhibit BVDV replication through the activation of the RIG-1/TBK-1 pathway to induce ISGs, and offering a novel therapeutic approach for combating BVDV. Thus, it is crucial to negatively regulate SREBP-1/FAS signaling molecules in order to create novel antiviral drugs that are safe, effective, and broad-spectrum.

Bis-Iridoid Glycosides and Triterpenoids from Kolkwitzia amabilis and Their Potential as Inhibitors of ACC1 and ACL.

A comprehensive phytochemical investigation of the twigs/leaves and flower buds of Kolkwitzia amabilis, a rare deciduous shrub native to China, led to the isolation of 39 structurally diverse compounds. These compounds include 11 iridoid glycosides (1-4 and 7-13), 20 triterpenoids (5, 6, and 14-31), and 8 phenylpropanoids (32-39). Among these, amabiliosides A (1) and B (2) represent previously undescribed bis-iridoid glycosides, while amabiliosides C (3) and D (4) feature a unique bis-iridoid-monoterpenoid indole alkaloid scaffold with a tetrahydro-β-carboline-5-carboxylic acid moiety. Amabiliacids A (5) and B (6) are 24-nor-ursane-type triterpenoids characterized by an uncommon ∆11,13(18) transannular double bond. Their chemical structures and absolute configurations were elucidated through spectroscopic data and electronic circular dichroism analyses. Compound 2 exhibited a moderate inhibitory effect against acetyl CoA carboxylase 1 (ACC1), with an IC50 value of 9.6 μM. Lonicejaposide C (8), 3β-O-trans-caffeoyl-olean-12-en-28-oic acid (29), and (23E)-coumaroylhederagenin (31) showed notable inhibitory effects on ATP-citrate lyase (ACL), with IC50 values of 3.6, 1.6, and 4.7 μM, respectively. Additionally, 3β-acetyl-ursolic acid (17) demonstrated dual inhibitory activity against both ACC1 and ACL, with IC50 values of 10.3 and 2.0 μM, respectively. The interactions of the active compounds with ACC1 and ACL enzymes were examined through molecular docking studies. From a chemotaxonomic perspective, the isolation of bis-iridoid glycosides in this study may aid in clarifying the taxonomic relationship between the genera Kolkwitzia and Lonicera within the Caprifoliaceae family. These findings highlight the importance of conserving plant species with unique and diverse secondary metabolites, which could serve as potential sources of new therapeutic agents for treating ACC1/ACL-associated diseases.

Molecular Interaction Between Vasopressin and Insulin in Regulation of Metabolism: Impact on Cardiovascular and Metabolic Diseases.

Numerous compounds involved in the regulation of the cardiovascular system are also engaged in the control of metabolism. This review gives a survey of literature showing that arginine vasopressin (AVP), which is an effective cardiovascular peptide, exerts several direct and indirect metabolic effects and may play the role of the link adjusting blood supply to metabolism of tissues. Secretion of AVP and activation of AVP receptors are regulated by changes in blood pressure and body fluid osmolality, hypoxia, hyperglycemia, oxidative stress, inflammation, and several metabolic hormones; moreover, AVP turnover is regulated by insulin. Acting on V1a receptors in the liver, AVP stimulates glycogenolysis, reduces synthesis of glycogen, and promotes fatty acid synthesis and acetyl CoA carboxylase activity. Stimulating V1b receptors in the pancreatic islands, AVP promotes release of insulin and glucagon-like peptide-1 (GLP-1) and potentiates stimulatory effects of glucose and ACTH on secretion of insulin. Simultaneously, insulin increases AVP secretion by neurons of the paraventricular nucleus and the supraoptic nucleus. There is strong evidence that secretion of AVP and its metabolic effectiveness are significantly altered in metabolic and cardiovascular diseases. Both experimental and clinical data indicate that inappropriate interactions of AVP and insulin play an important role in the development of insulin resistance in obesity and diabetes mellitus.

Haplotyped genome mapping and functional characterisation of a blueberry anthocyanin acetyltransferase (AAT) controlling the accumulation of acylated anthocyanins.

Blueberry has a diversity of anthocyanins that confer its characteristic, blue-coloured skin. Whilst most cultivars produce only anthocyanin glycosides, some can add aliphatic or aromatic groups to the sugar moiety to create acylated anthocyanins. Due to their enhanced stability, acylated anthocyanins represent an attractive breeding target in blueberry. In this study a haplotype-resolved assembly of a previously identified quantitative trait locus on chromosome 2 of 'Hortblue Petite' (Vaccinium corymbosum) was created to identify candidate anthocyanin acyltransferase genes. One full-length gene (VcAAT1a) was selected, based on qPCR expression profiling and transient expression in tobacco leaves and strawberry and blueberry fruit flesh. In all three systems VcAAT1a was able to produce a range of acylated anthocyanins in plantae. Recombinant VcAAT1a protein demonstrated that, while VcAAT1a was able to act on both anthocyanin 3-O-glucosides and 3-O-galactosides, it could only utilise acetyl CoA as an acyl donor. Protein modelling using AlphaFold suggested that this restricted range in acyl donors may be due to a spatially restricted sub-pocket in the acyl binding site of VvAAT1. Finally, LUC/REN promoter activation assays revealed that the VcAAT1a promoter was transactivated by the VcMYBPA1 and VcMYBPA2 transcription factors, further expanding our knowledge of anthocyanin regulation in blueberry.

The physiological role of gamma-aminobutyric acid in relieving the effect of furfural inhibitor for improvement the production of lipid in D. intermedius Z8.

In order to improve the production and quality of lipid of D. intermedius Z8 under the furfural stress environment, different exogenous γ-aminobutyric acid (GABA) addition strategies and their reasons for improving the fermentation performance were investigated. For this purpose, the effect of different concentrations of furfural on the production of biomass and lipid in D. intermedius Z8 was researched. The result shows that the growth of D. intermedius Z8 nearly stopped when 1.0 g/L furfural was adopted. However, when 1.5 mmol/L GABA was used, the highest biomass and lipid production of 4.56 g/L and 1.83 g/L were obtained, respectively, which were 36.53 % and 61.95 % higher compared to the control group (the normal development BG11 medium). Additionally, the changes in microalgal cell morphology were analyzed using the scanning electron microscope (SEM) technology, and the results suggest that GABA addition could significantly mitigate the toxic effects of excessive furfural by maintaining the integrity of the cell. Simultaneously, the activities of key enzymes involved in fatty acid synthesis, such as nitrate reductase (NR), phosphatidic acid phosphatase (PAP), acetyl CoA carboxylase (ACC), and fatty acid synthase (FAS), were enhanced under the optimal condition, and thus improving lipid production. According to the results in this study, the exogenous GABA addition strategy is a simple and effective approach to improve microbial tolerance and enhance its fermentation performance.

Biosynthesis-Encoded Lipogenic Acetyl-CoA Measurement Using NMR Reveals Glucose-Driven Lipogenesis and Glutamine's Alternative Roles in Kidney Cancer.

Fatty acid de novo synthesis (FADNS) is a critical process in lipogenesis that is characteristically altered in clear cell renal cell carcinoma (ccRCC), which is the major type of kidney cancer. An important challenge in studying the FADNS process has been the accurate measurement of cytosolic lipogenic acetyl-CoA (AcCoA), the precursor in FADNS, due to its compartmentalization within cells. Here, we describe a novel NMR-based method to decode the isotopic enrichment of lipogenic AcCoA, which, as we demonstrated, is encoded in the simple signal ratios of the geminal methyl groups of lanosterol during its biosynthesis. The approach was validated based on the independence of the tracer enrichment and species along with the expected FADNS modulation using differentially enriched tracers and a well-studied drug. Application of this technique to 786-O ccRCC cells showed that glucose may serve as a major carbon source for lipogenic AcCoA in FADNS at physiological nutrient concentrations, at odds with previous studies that indicated glutamine's dominant role through reductive carboxylation under higher nutrient conditions. Further investigation into glutamine's alternative roles in ccRCC cells suggested its major involvement in the bioenergetic TCA cycle, pyrimidine synthesis, and glutathione synthesis, which is also critical in ccRCC growth. The glutamine-dependent glutathione synthesis was also suggested as a possible metabolic vulnerability compared to normal kidney cells using a glutathione synthesis inhibitor. The current study provides a simple tool for studying an important aspect of lipid metabolism and suggests translational implications for targeting glucose-driven lipogenesis and glutamine-supported glutathione synthesis in ccRCC.

Boosted Meat Flavor by the Metabolomic Effects of Nile Tilapia Dietary Inclusion of Zophobas atratus Larval Meal.

Zophobas atratus larval meal (ZLM) is a high-quality feed supplement with potential activities that can improve fish growth performance and promote meat quality. However, there have been limited recent studies investigating the metabolic effects of ZLM. Therefore, this study aims to uncover the metabolomic mechanism through which ZLM improves tilapia meat flavor using metabolomic strategies. In this study, soybean meal in the basal diets was replaced with 15%, 30%, or 60% ZLM, where anti-nutrient factors were destroyed by high temperature treatment. After being fed these ZLM supplements for 30 days, dorsal muscles were collected from tilapia for meat sensory evaluation tests. Liver samples were also collected for metabolomic analysis using the gas chromatography-mass spectrometry (GC-MS) platform and combined with biochemical assays to verify metabolism-related enzyme activities and reveal crucial metabolic pathways and critical biomarkers associated with ZLM's ability to improve meat flavor. In tilapia livers, ZLM enhanced the activity of enzymes involved in energy metabolism including succinate dehydrogenase (SDH), pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (α-KGDH), NADP-malate dehydrogenase (NAD-MDH) and mitochondrial isocitrate dehydrogenase (ICDHm). This resulted in increased levels of reduced nicotinamide adenine dinucleotide (NADH), acetyl CoA and ATP which led to accumulation of flavor fatty acids such as arachidonic acid, linoleic acid (9,12-Octadecadienoic acid), linolenic acid (9,12,15-Octadecatrienoic acid) and oleic acid (9-Octadecenoic acid). Additionally, there was an increase in flavor nucleotides like guanosine adenosine-5'-monophosphate and uridine-5'-monophosphate while off-flavor metabolites like inosine and hypoxanthine decreased. Furthermore, beneficial metabolomic responses led to a decrease in off-flavor metabolites such as 2-methylisoborneol trimethylamine and geosmin while increasing umami metabolites like 2-methyl-3-furanthiol and nonanal. This metabolomic study demonstrates that inclusion of ZLM diets enhances the flavor profile of tilapia dorsal muscle. The accumulation of flavor compounds, coupled with a reduction in earthy taste and off-flavor metabolites, contributes to an improved meat flavor and freshness. Additionally, there is an increase in the levels of flavor-related amino acids and nucleotides. These previously unidentified metabolic effects highlight the potential significance of ZLM as a dietary supplement for enhancing the biosynthesis of flavor metabolites in tilapia.

Sensitivity of Southcentral U.S. Johnsongrass Accessions to Selected Herbicides

Abstract New technologies in grain sorghum allow for the use of multiple acetyl CoA carboxylase- (ACCase) or acetolactate synthase- (ALS) inhibiting herbicides for johnsongrass control. With the growing issue of herbicide resistance, producers need to understand which herbicides will successfully control johnsongrass accessions. To determine the efficacy of herbicides recently registered or ones with potential to become available for use in grain sorghum, johnsongrass seeds were collected from 2017 to 2021 in Arkansas, Kansas, Texas, and Oklahoma and were screened for sensitivity to fluazifop, quizalofop, nicosulfuron, and imazamox. Additionally, glyphosate sensitivity was evaluated because of its use before planting or postharvest. Quizalofop resulted in 100% mortality of all johnsongrass accessions. Of the johnsongrass accessions evaluated, 89% were completely controlled with glyphosate. The ALS inhibitors nicosulfuron and imazamox resulted in 100% mortality of all Oklahoma accessions, but failures occurred on samples from other states. One accession from Kansas, 12 from Texas, and eight from Arkansas were found to have reduced sensitivity to nicosulfuron and imazamox. If producers plan to plant grain sorghum in areas with johnsongrass populations, an ACCase-inhibitor herbicide will most likely provide effective control. Imazamox and nicosulfuron, in conjunction with the appropriate trait, can be utilized in areas with sensitive johnsongrass populations or where other sensitive grass species are present.

Mitochondria: the epigenetic regulators of ovarian aging and longevity.

Ovarian aging is a major health concern for women. Ovarian aging is associated with reduced health span and longevity. Mitochondrial dysfunction is one of the hallmarks of ovarian aging. In addition to providing oocytes with optimal energy, the mitochondria provide a co-substrate that drives epigenetic processes. Studies show epigenetic alterations, both nuclear and mitochondrial contribute to ovarian aging. Both, nuclear and mitochondrial genomes cross-talk with each other, resulting in two ways orchestrated anterograde and retrograde response that involves epigenetic changes in nuclear and mitochondrial compartments. Epigenetic alterations causing changes in metabolism impact ovarian function. Key mitochondrial co-substrate includes acetyl CoA, NAD+, ATP, and α-KG. Thus, enhancing mitochondrial function in aging ovaries may preserve ovarian function and can lead to ovarian longevity and reproductive and better health outcomes in women. This article describes the role of mitochondria-led epigenetics involved in ovarian aging and discusses strategies to restore epigenetic reprogramming in oocytes by preserving, protecting, or promoting mitochondrial function.

EXTH-64. MECHANISMS OF MITOCHONDRIAL COMPLEX-I SENSITIVITY IN GBM

Abstract The biguanide metformin that is widely used for the management of Type 2 diabetes is being evaluated as an anti-neoplastic agent in cancer trials including GBM (NCT02780024, NCT03243851). The mechanisms of action of metformin as an anti-neoplastic agent is still being investigated, but recent studies have unequivocally demonstrated that its therapeutic effects in tumor cell growth and signaling are dependent on inhibition of mitochondrial complex I. Based on the success of metformin in combination therapy in some cancers, new generation of mitochondrial complex I inhibitors (MCI-i) such as IM156, IACS-010759, are currently under investigation. Diffusion of metformin across the plasma membrane occurs slowly. Its entry is facilitated by two transporters OCT1 and OCT2 that are not well expressed outside the liver and kidney. This presents a problem due to its lack of accumulation in other tissues at therapeutic concentrations. Therefore, identification of tumor subtypes that respond better to lower concentrations of metformin may be leveraged for targeted metformin therapy. Metformin GBM trials may be unsatisfactory since there are no molecular markers to distinguish metformin responders from non-responders. Identification of molecular markers may greatly improve metformin and other MCI-i-based therapy in GBM. By examining TCGA data and experimental analysis, we have identified that PDHA, that catalyzes pyruvate to acetyl Co-A, is a predictive molecular marker for MCI-i sensitivity in GBM and potentially beyond GBM. PDHA expression varies widely in patient tumors, potentially due to differential methylation. We are performing joint pathway analysis by examining differentially expressed genes and stable isotope tracing studies to determine distinct metabolic signatures in the two subsets of GBM. We are also performing Classification And Regression Tree (CART) analysis on a cohort of sixty primary GBM lines and tissues to predict if PDHA expression provides a binary outcome for MCI-i sensitivity.

Muscle metabolism in response to oxidized fish oil feed in juvenile Nile tilapia

To investigate the effect of oxidized fish oil on fish muscle metabolic responses and flesh quality, Nile tilapia (Oreochromis niloticus) weighing 13.73 ± 0.31 g were fed two diets for 12 weeks: a fresh fish oil (FFO) and a highly oxidized fish oil (OFO) diet. The peroxide value of the FFO and OFO diets was 2.2 meq/kg and 120.6 meq/kg, respectively. The OFO diet resulted in a decrease in lower growth, muscularity, nutritional value of fatty acids in the muscle, and density of myofibers. In the OFO group, the mRNA expression levels of lipolysis genes lipoprotein lipase (lpl) and hormone - sensitive lipase(hsl) in the muscle were upregulated compared with the FFO group, while, the mRNA expression level of acetyl CoA carboxylase was downregulated. Additionally, under the positive ion mode, the levels of DHA deposition in phosphatidylcholine and phosphatidyl ethanolamine were reduced in the muscle of the OFO group compared to the FFO group. A total of 68 proteins were identified in the muscle, of which 42 were up-regulated and 26 were down-regulated. The KEGG pathway enrichment analysis revealed that the major pathways of the differentially abundant proteins were the Wnt signaling pathway, TGF-beta signaling pathway, and proteoglycans in cancer. In summary, the OFO diet negatively affected growth, muscularity, and nutritional value. It also inhibited the transformation of myofiber types and led to the apoptosis of myofibers.

Electrochemically Promoted Regioselective C3–H Trifluoro/Difluoromethylation of 2H-Indazoles at Room Temperature

AbstractA green and sustainable electrochemical approach is developed for the regioselective C3–H trifluoro/difluoromethylation of 2H-indazoles at room temperature. Relatively less expensive C-soft (+)/Ni-foam (–) electrodes are utilized to selectively functionalize the 2H-indazoles effectively by avoiding the use of any external oxidant and transition-metal salt. Moreover, along with the C3–H trifluoromethylation, for the very first time, direct C3–H difluoromethylation of 2-phenyl-2H-indazoles is accomplished. Diverse C3–H trifluoro/difluoromethylated 2H-indazoles having an array of functionalities are successfully synthesized in moderate to very good yields. As an application, a precursor of both an estrogen receptor ligand and an acetyl Co-A carboxylase inhibitor is synthesized. A plausible reaction mechanism is proposed based on control experiments and cyclic voltammetry studies.

Discovery and Characterization of BAY-184: A New Potent and Selective Acylsulfonamide-Benzofuran In Vivo-Active KAT6AB Inhibitor.

KAT6A and KAT6B genes are two closely related lysine acetyltransferases that transfer an acetyl group from acetyl coenzyme A (AcCoA) to lysine residues of target histone substrates, hence playing a key role in chromatin regulation. KAT6A and KAT6B genes are frequently amplified in various cancer types. In breast cancer, the 8p11-p12 amplicon occurs in 12-15% of cases, resulting in elevated copy numbers and expression levels of chromatin modifiers like KAT6A. Here, we report the discovery of a new acylsulfonamide-benzofuran series as a novel structural class for KAT6A/B inhibition. These compounds were identified through high-throughput screening and subsequently optimized using molecular modeling and cocrystal structure determination. The final tool compound, BAY-184 (29), was successfully validated in an in vivo proof-of-concept study.

Macrogenomic analysis of tolerance and degradation mechanisms of polycyclic aromatic hydrocarbon in carbon and nitrogen metabolic pathways and associated bacterial communities during endogenous partial denitrification

The effective production of NO2−-N through endogenous partial denitrification (EPD) provides a promising perspective for the broader adoption and application of anaerobic ammonia oxidation. However, the accumulation of polycyclic aromatic hydrocarbons (PAHs) in the environment may worsen the operational challenges of the EPD system. This study evaluated the resilience of the EPD system to the toxic impacts of phenanthrene (PHE) and anthracene (ANT) through macrogenomic analysis. A control group was maintained under identical conditions for comparison. The results revealed that PHE and ANT had a relatively minimal impact on NO3−-N transformation and organic matter removal but significantly affected PO43−-P removal and NO2−-N accumulation in the EPD process. The PHE system achieved a higher NO2−-N accumulation, with a maximum NO3−-N to NO2−-N conversion ratio of 90.08 %. In contrast, the ANT system exhibited higher efficiency in the PO43−-P removal, achieving a peak removal rate of 74.94 %. Macrogenomic analysis revealed that PAHs significantly enriched both denitrifying glycogen-accumulating organisms (including Candidatus_Competibacter) and denitrifying polyphosphate-accumulating organisms (such as Thauera, Candidatus_Contendobacter, and Candidatus_Accumulibacter). This enrichment stabilized these organisms, facilitating NO2−-N accumulation and PO43−-P removal. Metabolic pathway analysis indicated that PHE promoted the enrichment of NO3−-N reductase and inhibited NO2−-N reductase activity. However, ANT stimulated oxidative phosphorylation and the phosphate cycle. Moreover, PAH metabolites enhanced the expression of key genes encoding succinate dehydrogenase and isocitrate dehydrogenase in the tricarboxylic acid cycle within the EPD process, leading to increase the synthesis and utilization of acetyl coenzyme-A. Notably, significant differences were observed between the effects of PHE and ANT on these metabolic processes. This study provides new methods for treating PAH-containing wastewater through the combination of EPD and anaerobic ammonia oxidation.

Molecular mechanics studies of factors affecting overall rate in cascade reactions: Multi-enzyme colocalization and environment.

Millions of years of evolution have optimized many biosynthetic pathways by use of multi-step catalysis. In addition, multi-step metabolic pathways are commonly found in and on membrane-bound organelles in eukaryotic biochemistry. The fundamental mechanisms that facilitate these reaction processes provide strategies to bioengineer metabolic pathways in synthetic chemistry. Using Brownian dynamics simulations, here we modeled intermediate substrate transportation of colocalized yeast-ester biosynthesis enzymes on the membrane. The substrate acetate ion traveled from the pocket of aldehyde dehydrogenase to its target enzyme acetyl-CoA synthetase, then the substrate acetyl CoA diffused from Acs1 to the active site of the next enzyme, alcohol-O-acetyltransferase. Arranging two enzymes with the smallest inter-enzyme distance of 60 Å had the fastest average substrate association time as compared with anchoring enzymes with larger inter-enzyme distances. When the off-target side reactions were turned on, most substrates were lost, which suggests that native localization is necessary for efficient final product synthesis. We also evaluated the effects of intermolecular interactions, local substrate concentrations, and membrane environment to bring mechanistic insights into the colocalization pathways. The computation work demonstrates that creating spatially organized multi-enzymes on membranes can be an effective strategy to increase final product synthesis in bioengineering systems.