Carbohydrate Metabolism Research Articles

Effects of dietary carbohydrate levels on growth performance, feed utilization, liver histology and intestinal microflora of juvenile tiger puffer, Takifugu rubripes

An 8-week feeding trial was conducted to investigate the effects of dietary carbohydrate levels on growth performance, body composition, and plasma biochemical parameters, liver and gut histology, and intestinal microbiota of Takifugu rubripes. Five isonitrogenous and isolipidic experimental diets were formulated with graded levels of cassava starch: 0% (C0), 8% (C8), 16% (C16), 24% (C24), and 32% (C32). The results showed that weight gain rate, specific growth rate (SGR) and protein efficiency ratio significantly increased in C16 and C24 groups (P < 0.05), while the feed conversion ratio expressed an opposite trend. Feeding rate is decreased with the increasing of carbohydrate levels (P < 0.05). Liver lipid content was dramatically increased in C24 treatment (P < 0.05). The intestinal amylase activity and muscle glycogen content were higher than other groups (P < 0.05). Plasma glucose levels were increased in C24 and C32 groups (P < 0.05). In addition, the abundance of intestinal potentially pathogenic bacteria (Staphylococcus) decreased, and the intestinal probiotics (Lactobacillus) increased with the increasing of dietary carbohydrate level (P < 0.05). The intestinal bacteria, indicated by Sobs, Chao and Shannon indexes, were more diverse with the increasing of the carbohydrate level (P < 0.05), meanwhile the abundance of transport and metabolism of carbohydrate, lipid and amino acid were enhanced with the increasing of carbohydrate levels (P < 0.05). In conclusion, appropriate supplementation of carbohydrate in diet promoted growth performance and feed utilization of juvenile Takifugu rubripes, and increased the liver lipid and muscle glycogen content, as well as intestinal microbial diversity. Based on the quadratic regression between SGR and carbohydrate levels, the optimal dietary carbohydrate level of juvenile Takifugu rubripes is 21.6%.

Transcriptomic analysis provides insights into the molecular mechanism of melatonin-mediated cadmium tolerance in Medicago sativa L.

Cadmium (Cd), a toxic element, often makes a serious threat to plant growth and development. Previous studies found that melatonin (Mel) reduced Cd accumulation and reestablished the redox balance to alleviate Cd stress in Medicago sativa L., however, the complex molecular mechanisms are still elusive. Here, comparative transcriptome analysis and biochemical experiments were conducted to explore the molecular mechanisms of Mel in enhancing Cd tolerance. Results showed that 7237 differentially expressed genes (DEGs) were regulated by Mel pretreatment to Cd stress compared to the control condition in roots of Medicago sativa L. Besides, in comparison with Cd stress alone, Mel upregulated 1081 DEGs, and downregulated 1085 DEGs. These DEGs were mainly involved in the transcription and translation of genes and folding, sorting and degradation of proteins, carbohydrate metabolism, and hormone signal network. Application of Mel regulated the expression of several genes encoding ribosomal protein and E3 ubiquitin-protein ligase involved in folding, sorting and degradation of proteins. Moreover, transcriptomic analyse suggested that Mel might regulate the expression of genes encoding pectin lyase, UDP-glucose dehydrogenase, sucrose-phosphate synthase, hexokinase-1, and protein phosphorylation in the sugar metabolism. Therefore, these could promote sucrose accumulation and subsequently alleviate the Cd damage. In conclusion, above findings provided the mining of important genes and molecular basis of Mel in mitigating Cd tolerance and genetic cultivation of Medicago sativa L.

Nitric oxide alleviates programmed cell death induced by cadmium in Solanum lycopersicum seedlings through protein S-nitrosylation

Cadmium (Cd), as a non-essential and toxic heavy metal in plants, has deleterious effects on plant physiological and biochemical processes. Nitric oxide (NO) is one of the most important signaling molecules for plants to response diverse stresses. Here, we found that Cd-induced programmed cell death (PCD) was accompanied by NO bursts, which exacerbated cell death when NO was removed and vice versa. Proteomic analysis of S-nitrosylated proteins showed that the differential proteins in Cd-induced PCD and in NO-alleviated PCD mainly exist together in carbohydrate metabolism and amino acid metabolism, while some of the differential proteins exist alone in metabolism of cofactors and vitamins and lipid metabolism. Meanwhile, S-nitrosylation of proteins in porphyrin and chlorophyll metabolism and starch and sucrose metabolism could explain the leaf chlorosis induced by PCD. Moreover, protein transport protein SEC23, ubiquitinyl hydrolase 1 and pathogenesis-related protein 1 were identified to be S-nitrosylated in vivo, and their expressions were increased in Cd-induced PCD while decreased in NO treatment. Similar results were obtained in tomato seedlings with higher S-nitrosylation. Taken together, our results indicate that NO might be involved in the regulation of Cd-induced PCD through protein S-nitrosylation, especially proteins involved in PCD response.

Nutritional and Herbal Supplements for Weight Loss Management: A Review

Abstract: Millions of dollars have been spent on nutritional and herbal supplements, which are used by many overweight people to lose weight. However, there are concerns about the effectiveness and safety of several supplements. By boosting energy expenditure, satiety, fat oxidation, inhibiting dietary fat absorption, moderating carbohydrate metabolism, raising fat excretion, increasing water elimination, and improving mood, supplements are thought to aid in weight reduction. This study provides an overview of the present status of the research on a few particular dietary supplements, herbal combinations, and combination products, as this field is evolving quickly. More studies on numerous supplements are required to verify their efficacy because they have not been examined in randomised controlled trials. Nutritionists and medical experts should go through dietary supplement use with patients and notify the FDA of any negative side effects. Additional rules would be required for the manufacture, sale, and marketing of these supplements.

The Application Timing of a Cytokinin B-Mo-based Product Affects the Characteristics of Rooted Cuttings and Nonstructural Carbohydrates of Coleus (Plectranthus scutellarioides cv. Wild Lime) during Adventitious Root Development

A large group of horticulture species are propagated vegetatively through shoot-tip cuttings harvested from stock plants and planted to form adventitious roots. Adventitious rooting leads to establishing a carbohydrate sink in the region of root regeneration that is highly dependent on energy and carbon skeletons. We hypothesized that the timing of exogenous applications of cytokinin (CK) and boron (B)–molybdenum (Mo)-based products during adventitious root development can affect the flow of sugars from leaves to sinks, carbon allocation to the adventitious roots, and the quality of rooted cuttings. During this project, we aimed to determine if the application time of a CK/B-Mo-based product during the adventitious root development of unrooted cuttings would impact the source-to-sink relationship and, hence, affect plug growth and quality. A sink-strengthening commercial product based on cytokinin, B, and Mo was applied at four plug development stages plus a negative control as follows: T1, plants without product (control); T2, sticking stage (starting 24 hours after the sticking); T3, callus formation stage; T4, root development stage; and T5, toning stage. The root and shoot lengths and dry matter, number of leaves, leaf chlorophyll content, root-to-shoot ratio (based on dry matter), and nonstructural carbohydrate contents were measured. The timing of the application of the product impacted the root development, quality of the cuttings, and nonstructural carbohydrate content. Product application during the adventitious root dedifferentiation and induction phases (T2) resulted in the shortest root and shoot lengths, lowest dry matter accumulation, lowest nonstructural carbohydrate contents, and some phytotoxicity. Application during the initiation phase (T3) resulted in greater root length, total dry matter, and total soluble sugar contents compared with the control. Application during the expression phase (T4) resulted in the largest root length and mass and the highest sucrose contents. Applying the product when the roots had grown and reached all the edges of the growing media (T5) did not have any benefits compared with the control. This study provides new insights into the application timing of exogenous CKs, B, and Mo to generate a well-toned rooted coleus cutting and potential explanations in relation to nonstructural carbohydrate metabolism.

Oral microbiome sequencing revealed the enrichment of Fusobacterium sp., Porphyromonas sp., Campylobacter sp., and Neisseria sp. on the oral malignant fibroma surface of giant panda

IntroductionMicrobial community composition is closely associated with host disease onset and progression, underscoring the importance of understanding host–microbiota dynamics in various health contexts.MethodsIn this study, we utilized full-length 16S rRNA gene sequencing to conduct species-level identification of the microorganisms in the oral cavity of a giant panda (Ailuropoda melanoleuca) with oral malignant fibroma.ResultsWe observed a significant difference between the microbial community of the tumor side and non-tumor side of the oral cavity of the giant panda, with the latter exhibiting higher microbial diversity. The tumor side was dominated by specific microorganisms, such as Fusobacterium simiae, Porphyromonas sp. feline oral taxon 110, Campylobacter sp. feline oral taxon 100, and Neisseria sp. feline oral taxon 078, that have been reported to be associated with tumorigenic processes and periodontal diseases in other organisms. According to the linear discriminant analysis effect size analysis, more than 9 distinct biomarkers were obtained between the tumor side and non-tumor side samples. Furthermore, the Kyoto Encyclopedia of Genes and Genomes analysis revealed that the oral microbiota of the giant panda was significantly associated with genetic information processing and metabolism, particularly cofactor and vitamin, amino acid, and carbohydrate metabolism. Furthermore, a significant bacterial invasion of epithelial cells was predicted in the tumor side.DiscussionThis study provides crucial insights into the association between oral microbiota and oral tumors in giant pandas and offers potential biomarkers that may guide future health assessments and preventive strategies for captive and aging giant pandas.

Elevated Temperature Effects on Protein Turnover Dynamics in Arabidopsis thaliana Seedlings Revealed by 15N-Stable Isotope Labeling and ProteinTurnover Algorithm

Global warming poses a threat to plant survival, impacting growth and agricultural yield. Protein turnover, a critical regulatory mechanism balancing protein synthesis and degradation, is crucial for the cellular response to environmental changes. We investigated the effects of elevated temperature on proteome dynamics in Arabidopsis thaliana seedlings using 15N-stable isotope labeling and ultra-performance liquid chromatography-high resolution mass spectrometry, coupled with the ProteinTurnover algorithm. Analyzing different cellular fractions from plants grown under 22 °C and 30 °C growth conditions, we found significant changes in the turnover rates of 571 proteins, with a median 1.4-fold increase, indicating accelerated protein dynamics under thermal stress. Notably, soluble root fraction proteins exhibited smaller turnover changes, suggesting tissue-specific adaptations. Significant turnover alterations occurred with redox signaling, stress response, protein folding, secondary metabolism, and photorespiration, indicating complex responses enhancing plant thermal resilience. Conversely, proteins involved in carbohydrate metabolism and mitochondrial ATP synthesis showed minimal changes, highlighting their stability. This analysis highlights the intricate balance between proteome stability and adaptability, advancing our understanding of plant responses to heat stress and supporting the development of improved thermotolerant crops.

The Impact of Lactobacillus delbrueckii Hepatic Metabolism in Post-Weaning Piglets

Lactobacillus delbrueckii garners interest for its contributions to gut microecological balance, diarrheal prevention and treatment, immune modulation, growth promotion, and meat quality enhancement in livestock. However, its impact on the gut microbiota and liver metabolism in weaned piglets is less documented. This study involved 80 Duroc-Landrace-Yorkshire weaned piglets aged 28 days, randomized into two groups with four replicates each and ten piglets per replicate. Over a 28-day period, the piglets were fed either a basal diet (control group) or the same diet supplemented with 0.1% Lactobacillus delbrueckii microcapsules (≥1.0 × 1010 CFU/g) (Lactobacillus delbrueckii group). The principal findings are as follows: During the initial phase of the experiment, supplementation with Lactobacillus delbrueckii increased the levels of L-phenylalanine and L-lysine in the liver while reducing the L-alanine levels, thereby enhancing the aminoacyl–tRNA synthesis pathway in weaned piglets. In the later phase, Lactobacillus delbrueckii supplementation boosted the liver arachidonic acid content, strengthening the arachidonic acid metabolic pathway in the piglets. The gut microbiota and their metabolites likely play a role in regulating these processes. These results indicate that, compared to the control group, Lactobacillus delbrueckii reduced weaning stress-induced liver damage and metabolic disorders, increased liver glycogen content, and enhanced liver antioxidant function by improving the metabolism of lipids and carbohydrates. Consequently, the liver functioned more healthily.

Obesity and gestation in Criollo mares: endocrine and metabolic profiles

The aim of this study was to determine the effect of body condition score (BCS) on metabolic and endocrine parameters in pregnant Criollo mares (n=41), which were categorized according to their BCS as obese (7 to 9 BCS, n=26) or normal (5 to 7, n=15). Blood samples were taken during gestation in 3 periods: between 3.5 and 5 months (I), 8 and 9 months (II) and in the last month of gestation (III). The data was analyzed in the statistical model by mixed procedures, including BCS, gestational period and their interaction as fixed effects. BCS was only different in period I, as normal mares increased their BCS in the later periods. Leptin concentrations were greater in obese mares when compared to non-obese mares during all sampling periods (P<0.01), while glucose concentrations were also greater in the former group (P<0.01) but only during the first sampling period. Insulin concentrations and homeostatic model assessment for insulin resistance (HOMA-IR) index were greater also in obese mares in periods I and III (P<0.05). Adiponectin concentrations in period I were lower in obese mares (P<0.05). Cholesterol concentrations increased during gestation, and obese mares tended to have greater concentrations than nonobese mares (P<0.1). Triglyceride concentrations were not affected by group or gestational period. This study revealed adaptations in carbohydrate and lipid metabolism during gestation in mares. Several parameters are dependent on the degree of body fat reserves, which are reflected in the concentrations of biomarkers such as leptin and adiponectin. Insulin concentration in obese mares was higher than non-obese mares at the end of gestation, a similar profile was observed for HOMA-IR although cutoff values are yet still to be validated.

Proteomic Analysis of Milk Fat Globule Membrane Protein Modulation of Differently Expressed Proteins in Lactobacillus plantarum under Bile Salt Stress.

Tolerance to bile stress is a crucial property for lactic acid bacteria (LAB) to survive in the gastrointestinal tract and exert their beneficial effects. Whey powder enriched with milk fat globule membrane proteins (M-WPI) as a functional component is protective for strains under stress conditions. The current study investigated the key mechanisms of action involved in Lactobacillus plantarum (L. plantarum) CGMCC 23701 survival in the presence of bile and the protective mechanism of M-WPI. According to proteomic analysis (proteomics), there could be several reasons for the greater protective effect of M-WPI. These include promoting the synthesis of fatty acids and peptidoglycans to repair the structure of the cell surface, regulating the metabolism of carbohydrates and amino acids to release energy and produce a range of precursors, enabling the expression of the repair system to repair damaged DNA, and promoting the expression of proteins associated with the multidrug efflux pump, which facilitates the exocytosis of intracellular bile salts. This study helps us to better understand the changes in proteome of L. plantarum CGMCC 23701 under bile salt stress and M-WPI protection, which will provide a new method for the protection and development of functional LAB.

Zinc regulation of chlorophyll fluorescence and carbohydrate metabolism in saline-sodic stressed rice seedlings.

Saline-sodic stress can limit the absorption of available zinc in rice, subsequently impacting the normal photosynthesis and carbohydrate metabolism of rice plants. To investigate the impact of exogenous zinc application on photosynthesis and carbohydrate metabolism in rice grown in saline-sodic soil, this study simulated saline-sodic stress conditions using two rice varieties, 'Changbai 9' and 'Tonghe 899', as experimental materials. Rice seedlings at 4weeks of age underwent various treatments including control (CT), 2μmol·L-1 zinc treatment alone (Z), 50mmol·L-1 saline-sodic treatment (S), and 50mmol·L-1 saline-sodic treatment with 2μmol·L-1 zinc (Z + S). We utilized JIP-test to analyze the variations in excitation fluorescence and MR820 signal in rice leaves resulting from zinc supplementation under saline-sodic stress, and examined the impact of zinc supplementation on carbohydrate metabolism in both rice leaves and roots under saline-sodic stress. Research shows that zinc increased the chloroplast pigment content, specific energy flow, quantum yield, and performance of active PSII reaction centers (PIABS), as well as the oxidation (VOX) and reduction rate (Vred) of PSI in rice leaves under saline-sodic stress. Additionally, it decreased the relative variable fluorescence (WK and VJ) and quantum energy dissipation yield (φDO) of the rice. Meanwhile, zinc application can reduce the content of soluble sugars and starch in rice leaves and increasing the starch content in the roots. Therefore, the addition of zinc promotes electron and energy transfer in the rice photosystem under saline-sodic stress. It enhances rice carbohydrate metabolism, improving the rice plants' ability to withstand saline-sodic stress and ultimately promoting rice growth and development.

A MYB-related transcription factor ZmMYBR29 is involved in grain filling

BackgroundThe endosperm serves as the primary source of nutrients for maize (Zea mays L.) kernel embryo development and germination. Positioned at the base of the endosperm, the transfer cells (TCs) of the basal endosperm transfer layer (BETL) generate cell wall ingrowths, which enhance the connectivity between the maternal plant and the developing kernels. These TCs play a crucial role in nutrient transport and defense against pathogens. The molecular mechanism underlying BETL development in maize remains unraveled.ResultsThis study demonstrated that the MYB-related transcription factor ZmMYBR29, exhibited specific expression in the basal cellularized endosperm, as evidenced by in situ hybridization analysis. Utilizing the CRISPR/Cas9 system, we successfully generated a loss-of-function homozygous zmmybr29 mutant, which presented with smaller kernel size. Observation of histological sections revealed abnormal development and disrupted morphology of the cell wall ingrowths in the BETL. The average grain filling rate decreased significantly by 26.7% in zmmybr29 mutant in comparison to the wild type, which impacted the dry matter accumulation within the kernels and ultimately led to a decrease in grain weight. Analysis of RNA-seq data revealed downregulated expression of genes associated with starch synthesis and carbohydrate metabolism in the mutant. Furthermore, transcriptomic profiling identified 23 genes that expressed specifically in BETL, and the majority of these genes exhibited altered expression patterns in zmmybr29 mutant.ConclusionsIn summary, ZmMYBR29 encodes a MYB-related transcription factor that is expressed specifically in BETL, resulting in the downregulation of genes associated with kernel development. Furthermore, ZmMYBR29 influences kernels weight by affecting the grain filling rate, providing a new perspective for the complementation of the molecular regulatory network in maize endosperm development.

Genetic predisposition to metabolic dysfunction-associated fatty liver disease

The literature review highlights the issue of genetic risk factors associated with the development of metabolic dysfunction-associated fatty liver disease. Human genetic examinations revealed 132 genes among which 32 loci are strongly associated with the pathogenesis of metabolic dysfunction-associated fatty liver disease. It has been found that the risk of developing metabolic dysfunction-associated fatty liver disease is carried by single-nucleotide variants of various genes whose products are involved in lipid and carbohydrate metabolism, maintenance of the redox state, the development of inflammation and fibrosis of liver tissue, which are components of metabolic dysfunction-associated fatty liver disease reactome. The authors presented a detailed list of genetic factors singling out those that influence the risk of metabolic dysfunction-associated fatty liver disease and directly metabolic dysfunction-associated steatohepatitis and liver fibrosis. Also, they emphasized that it is the single-nucleotide variants of the genes of protein 3 containing a patatin-like phospholipase domain, transmembrane 6 superfamily member 2, and 17b-hydroxysteroid dehydrogenase type 13 that are characte­rized by the highest degree of association with metabolic dysfunction-associated fatty liver disease (odds ratio &gt; 1.6) compared to single-nucleotide variants of other genes identified by gene association studies. The combination of several polymorphisms increases the risk of development and severity of metabolic dysfunction-associated fatty liver disease. The additive steatogenic effect of protein 3 single-nucleotide gene variants containing a patatin-like phospholipase domain and transmembrane 6 superfamily member 2 is probably due to an increased expression of genes involved in de novo lipogenesis. The authors emphasize the need for genetic risk assessment of metabolic dysfunction-associated fatty liver disease, which should include molecular genetic testing at an early stage of examination.

The effect of pre-, pro- and synbiotics on the carbohydrate and lipid metabolism

The effect of pre-, pro- and synbiotics on the carbohydrate and lipid metabolism

Pituitary metastasis from lung adenocarcinoma: A case report

BACKGROUND Pituitary gland metastasis is an unusual event, and pituitary metastasis from lung adenocarcinoma is extremely rare and associated with poor prognosis. To date, approximately 15 cases have been reported. CASE SUMMARY Here, we present the case of a 64-year-old woman with pituitary metastasis derived from lung adenocarcinoma, which was difficult to distinguish from other sellar tumors. The patient presented to the neurosurgery clinic with blurred vision and intermittent headache. During hospitalization, brain computed tomography (CT) and magnetic resonance imaging revealed a pituitary macroadenoma. Chest CT revealed irregular nodules in the basal segment of the lower lobe of the left lung, which were likely lung cancer. Positron emission tomography-CT revealed a carbohydrate metabolism tumor in the lungs and sellar region, which was considered malignant. Postoperative pathological examination of the sellar tumor revealed lung adenocarcinoma. CONCLUSION Excision of pituitary metastases combined with radiotherapy and chemotherapy should be a priority treatment for patients with pituitary metastasis.

Upregulated Palmitoleate and Oleate Production in Escherichia coli Promotes Gentamicin Resistance

Metabolic reprogramming mediates antibiotic efficacy. However, metabolic adaptation of microbes evolving from antibiotic sensitivity to resistance remains undefined. Therefore, untargeted metabolomics was conducted to unveil relevant metabolic reprogramming and potential intervention targets involved in gentamicin resistance. In total, 61 metabolites and 52 metabolic pathways were significantly altered in gentamicin-resistant E. coli. Notably, the metabolic reprogramming was characterized by decreases in most metabolites involved in carbohydrate and amino acid metabolism, and accumulation of building blocks for nucleotide synthesis in gentamicin-resistant E. coli. Meanwhile, fatty acid metabolism and glycerolipid metabolism were also significantly altered in gentamicin-resistant E. coli. Additionally, glycerol, glycerol-3-phosphate, palmitoleate, and oleate were separately defined as the potential biomarkers for identifying gentamicin resistance in E. coli. Moreover, palmitoleate and oleate could attenuate or even abolished killing effects of gentamicin on E. coli, and separately increased the minimum inhibitory concentration of gentamicin against E. coli by 2 and 4 times. Furthermore, palmitoleate and oleate separately decreased intracellular gentamicin contents, and abolished gentamicin-induced accumulation of reactive oxygen species, indicating involvement of gentamicin metabolism and redox homeostasis in palmitoleate/oleate-promoted gentamicin resistance in E. coli. This study identifies the metabolic reprogramming, potential biomarkers and intervention targets related to gentamicin resistance in bacteria.

The bacterial patterns suggesting the dynamic features of tick-associated microorganisms in hard ticks

BackgroundTicks are blood-feeding significant arthropods that can harbour various microorganisms, including pathogens that pose health risks to humans and animals. Tick-symbiont microorganisms are believed to influence tick development, but the intricate interactions between these microbes and the relationships between different tick-borne microorganisms remain largely unexplored.ResultsBased on 111 tick pool samples presenting questing and engorged statuses including 752 questing tick and 1083 engorged tick from cattle and goats, which were collected in two types of geographic landscape (semi-desert and alpine meadow). We observed significant variations in the composition of tick-borne microorganisms across different environments and blood-engorgement statuses, with a pronounced divergence in symbionts compared to environmental bacteria. Metabolic predictions revealed over 90 differential pathways for tick-borne microorganisms in distinct environments and more than 80 metabolic variations in response to varying blood engorgement statuses. Interestingly, nine pathways were identified, particularly related to chorismate synthesis and carbohydrate metabolism. Moreover, microbial network relationships within tick-borne microorganism groups were highly distinct across different environments and blood-engorgement statuses. The microbial network relationships of symbionts involve some pathogenic and environmental microorganisms. Regression modelling highlighted positive correlations between the Coxiella symbiont and related pathogens, while some environmental bacteria showed strong negative correlations with Coxiella abundance. We also identified commensal bacteria/pathogens in bacterial cooccurrence patterns. Furthermore, we tested pathogenic microorganisms of each tick sample analysis revealed that 86.36% (1601/1855) of the tick samples carried one or more pathogenic microorganisms, The total carrier rate of bacterial pathogens was 43.77% ((812/1855). Most blood samples carried at least one pathogenic microorganism. The pathogens carried by the ticks have both genus and species diversity, and Rickettsia species are the most abundant pathogens among all pathogens.ConclusionOur findings underscore that the bacterial pattern of ticks is dynamic and unstable, which is influenced by the environment factors and tick developmental characteristics.

Characterising 24-h skeletal muscle gene expression alongside metabolic & endocrine responses under diurnal conditions.

Skeletal muscle plays a central role in the storage, synthesis, and breakdown of nutrients, yet little research has explored temporal responses of this human tissue, especially with concurrent measures of systemic biomarkers of metabolism. To characterise temporal profiles in skeletal muscle expression of genes involved in carbohydrate metabolism, lipid metabolism, circadian clocks, and autophagy and descriptively relate them to systemic metabolites and hormones during a controlled laboratory protocol. Ten healthy adults (9M/1F, mean ± SD: age: 30 ± 10 y; BMI: 24.1 ± 2.7 kg·m-2) rested in the laboratory for 37 hours with all data collected during the final 24 hours of this period (i.e., 0800-0800 h). Participants ingested hourly isocaloric liquid meal replacements alongside appetite assessments during waking before a sleep opportunity from 2200-0700 h. Blood samples were collected hourly for endocrine and metabolite analyses, with muscle biopsies occurring every 4 h from 1200 h to 0800 h the following day to quantify gene expression. Plasma insulin displayed diurnal rhythmicity peaking at 1804 h. Expression of skeletal muscle genes involved in carbohydrate metabolism (Name - Acrophase; GLUT4 - 1440 h; PPARGC1A -1613 h; HK2 - 1824 h) and lipid metabolism (FABP3 - 1237 h; PDK4 - 0530 h; CPT1B - 1258 h) displayed 24 h rhythmicity that reflected the temporal rhythm of insulin. Equally, circulating glucose (0019 h), NEFA (0456 h), glycerol (0432 h), triglyceride (2314 h), urea (0046 h), CTX (0507 h) and cortisol concentrations (2250 h) also all displayed diurnal rhythmicity. Diurnal rhythms were present in human skeletal muscle gene expression as well systemic metabolites and hormones under controlled diurnal conditions. The temporal patterns of genes relating to carbohydrate and lipid metabolism alongside circulating insulin are consistent with diurnal rhythms being driven in part by the diurnal influence of cyclic feeding and fasting.

The basement membrane regulates the cellular localization and the cytoplasmic interactome of Yes-Associated Protein (YAP) in mammary epithelial cells.

The Hippo pathway, a signaling cascade involved in the regulation of organ size and several other processes, acts as a conduit between extracellular matrix (ECM) cues and cellular responses. We asked whether the basement membrane (BM), a specialized ECM component known to induce quiescence and differentiation in mammary epithelial cells, would regulate the localization, activity, and interactome of YAP, a Hippo pathway effector. To address this question, we used a broad range of experimental approaches, including 2D and 3D cultures of both mouse and human mammary epithelial cells, as well as the developing mouse mammary gland. In contrast to malignant cells, nontumoral cells cultured with a reconstituted BM (rBM) displayed higher concentrations of YAP in the cytoplasm. Incidentally, when in the nucleus of rBM-treated cells, YAP resided preferentially at the nuclear periphery. In agreement with our cell culture experiments, YAP exhibited cytoplasmic predominance in ductal cells of developing mammary epithelia, where a denser BM is found. Conversely, terminal end bud (TEB) cells with a thinner BM displayed higher nucleus-to-cytoplasm ratios of YAP. Bioinformatic analysis revealed that genes regulated by YAP were overrepresented in the transcriptomes of microdissected TEBs. Consistently, mouse epithelial cells exposed to the rBM expressed lower levels of YAP-regulated genes, although the protein level of YAP and Hippo components were slightly altered by the treatment. Mass spectrometry analysis identified a differential set of proteins interacting with YAP in cytoplasmic fractions of mouse epithelial cells in the absence or presence of rBM. In untreated cells, YAP interactants were enriched in processes related to ubiquitin-mediated proteolysis, whereas in cells exposed to rBM YAP interactants were mainly key proteins related to amino acid, amino sugar, and carbohydrate metabolism. Collectively, we unraveled that the BM induces YAP translocation or retention in the cytoplasm of nontumoral epithelial cells and that in the cytoplasm YAP seems to undertake novel functions in metabolic pathways.

Comparative Metabolome Profiling for Revealing the Effects of Different Cooking Methods on Glutinous Rice Longjing57 (Oryza sativa L. var. Glutinosa)

Glutinous rice (GR), an important food crop in Asia, provides prolonged energy for the human body due to its high amylopectin content. The non-volatile metabolites generated by different cooking methods that affect the nutritional value and color of GR are still poorly understood. Herein, a widely targeted metabolomics approach was used to understand the effects of different cooking methods (steaming, baking, and frying) on the metabolite profiles of GR. Compared with other treatments, steamed GR had a brighter color and significantly lower contents of total sugar, starch, amylopectin, and amylose, at 40.74%, 14.13%, 9.78%, and 15.18%, respectively. Additionally, 70, 108, and 115 metabolites were significantly altered in the steaming, baking, and frying groups respectively, and amino acid and carbohydrate metabolism were identified as the representative metabolic pathways based on KEGG annotations. Further evaluation of 14 amino acids and 12 carbohydrates in steamed GR, especially 4-aminobutyric acid, suggested its high nutraceutical value. Additionally, multivariate analysis indicated that total sugar content, amylose content, beta-alanine methyl ester hydrochloride, and 4-aminobutyric acid played a critical role in color formation in raw and cooked GR. Finally, the levels of major amino acids and carbohydrates were quantified by conventional methods to verify the reliability of the metabolome. Consequently, this in-depth understanding of metabolite profiling in normal cooking methods has provided a foundation for the processing of GR products.