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At least 595 records · Page 33

Structural Similarities and Overlapping Activities among Dihydroflavonol 4-Reductase, Flavanone 4-Reductase, and Anthocyanidin Reductase Offer Metabolic Flexibility in the Flavonoid Pathway

Flavonoids are potent antioxidants that play a role in defense against pathogens, UV-radiation, and the detoxification of reactive oxygen species. Dihydroflavonol 4-reductase (DFR) and flavanone 4-reductase (FNR) reduce dihydroflavonols and flavanones, respectively, using NAD(P)H to produce flavan-(3)-4-(di)ols in flavonoid biosynthesis. Anthocyanidin reductase (ANR) reduces anthocyanidins to flavan-3-ols. In addition to their sequences, the 3D structures of recombinant DFR, FNR and ANR from sorghum and switchgrass showed a high level of similarity. The catalytic mechanism, substrate-specificity and key residues of three reductases were deduced from crystal structures, site-directed mutagenesis, molecular docking, kinetics, and thermodynamic ana-lyses. Although DFR displayed its highest activity against dihydroflavonols, it also showed activity against flavanones and anthocyanidins. It was inhibited by the flavonol quercetin and high concentrations of dihydroflavonols/flavonones. SbFNR1 and SbFNR2 did not show any activity against dihydroflavonols. However, SbFNR1 displayed activity against flavanones and ANR activity against two anthocyanidins, cyanidin and pelargonidin. Therefore, SbFNR1 and SbFNR2 could be specific ANR isozymes without delphinidin activity. Sorghum has high concentrations of 3-deoxyanthocyanidins in vivo, supporting the observed high activity of SbDFR against flavonols. Mining of expression data indicated substantial induction of these three reductase genes in both switchgrass and sorghum in response to biotic stress. Key signature sequences for proper DFR/ANR classification are proposed and could form the basis for future metabolic engineering of flavonoid metabolism.

3-deoxyanthocyanidin↗

Xanthohumol Alters Gut Microbiota Metabolism and Bile Acid Dynamics in Gastrointestinal Simulation Models of Eubiotic and Dysbiotic States

Xanthohumol (XN), a polyphenol from hops (Humulus lupulus), exhibits antioxidant, anti-inflammatory, antihyperlipidemic, and chemo-preventive activity. Preclinical evidence suggests gut microbiota are critical to mediating some of these bioactivities. Nevertheless, its precise impact on human gut microbiota, particularly at supplemental doses, remains poorly characterized. We evaluated 200 mg/day XN for 3 weeks on human gut microbiota in a eubiotic and dysbiotic model using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME®). Functional assessments of microbiota included quantification of XN metabolites, short-chain fatty acids (SCFAs), and untargeted metabolomics of the digestive metabolome. Bacterial composition was assessed by 16S rRNA gene sequencing. XN reduced alpha-diversity and short-chain fatty acid production in both models, as well as altered taxa abundance variably between models. XN disrupted bile acid metabolism through inhibition of microbial bile salt hydrolase (BSH). The modulation of bile acid metabolism has important implications for host-level bioactivity of XN.

Humulus lupulus↗

Untargeted GC-MS Metabolic Profiling of Anaerobic Gut Fungi Reveals Putative Terpenoids and Strain-Specific Metabolites

Background/Objectives: Anaerobic gut fungi (Neocallimastigomycota) are biotechnologically relevant, lignocellulose-degrading microbes with under-explored biosynthetic potential for secondary metabolites. Untargeted metabolomic profiling with gas chromatography–mass spectrometry (GC-MS) was applied to two gut fungal strains, Anaeromyces robustus and Caecomyces churrovis, to establish a foundational metabolomic dataset to identify metabolites and provide insights into gut fungal metabolic capabilities. Methods: Gut fungi were cultured anaerobically in rumen-fluid-based media with a soluble substrate (cellobiose), and metabolites were extracted using the Metabolite, Protein, and Lipid Extraction (MPLEx) method, enabling metabolomic and proteomic analysis from the same cell samples. Samples were derivatized and analyzed via GC-MS, followed by compound identification by spectral matching to reference databases, molecular networking, and statistical analyses. Results: Distinct metabolites were identified between A. robustus and C. churrovis, including 2,3-dihydroxyisovaleric acid produced by A. robustus and maltotriitol, maltotriose, and melibiose produced by C. churrovis. C. churrovis may polymerize maltotriose to form an extracellular polysaccharide, like pullulan. GC-MS profiling potentially captured sufficiently volatile products of proteomically detected, putative non-ribosomal peptide synthetases and polyketide synthases of A. robustus and C. churrovis. The triterpene squalene and triterpenoid tetrahymanol were putatively identified in A. robustus and C. churrovis. Their conserved, predicted biosynthetic genes—squalene synthase and squalene tetrahymanol cyclase—were identified in A. robustus, C. churrovis, and other anaerobic gut fungal genera. Conclusions: This study provides a foundational, untargeted metabolomic dataset to unmask gut fungal metabolic pathways and biosynthetic potential and to prioritize future efforts for compound isolation and identification.

Biochemistry & Molecular Biology↗

Reconfiguring Plant Metabolism for Biodegradable Plastic Production

For decades, plants have been the subject of genetic engineering to synthesize novel, value-added compounds. Polyhydroxyalkanoates (PHAs), a large class of biodegradable biopolymers naturally synthesized in eubacteria, are among the novel products that have been introduced to make use of plant acetyl-CoA metabolic pathways. It was hoped that renewable PHA production would help address environmental issues associated with the accumulation of nondegradable plastic wastes. However, after three decades of effort synthesizing PHAs, and in particular the simplest form polyhydroxybutyrate (PHB), and seeking to improve their production in plants, it has proven very difficult to reach a commercially profitable rate in a normally growing plant. This seems to be due to the growth defects associated with PHA production and accumulation in plant cells. Here, we review major breakthroughs that have been made in plant-based PHA synthesis using traditional genetic engineering approaches and discuss challenges that have been encountered. Then, from the point of view of plant synthetic biology, we provide perspectives on reprograming plant acetyl-CoA pathways for PHA production, with the goal of maximizing PHA yield while minimizing growth inhibition. Specifically, we suggest genetic elements that can be considered in genetic circuit design, approaches for nuclear genome and plastome modification, and the use of multiomics and mathematical modeling in understanding and restructuring plant metabolic pathways.

59 BASIC BIOLOGICAL SCIENCES↗

Breathing metabolic simulator

The development of a breathing metabolic simulator (BMS) is reported. This BMS simulates all of the breathing and metabolic parameters required for complete evaluation and test of life support and resuscitation equipment. It is also useful for calibrating and validating mechanical and gaseous pulmonary function test procedures. Breathing rate, breathing depth, breath velocity contour, oxygen uptake, and carbon dioxide release are all variable over wide ranges simulating conditions from sleep to hard work with respiratory exchange ratios covering the range from hypoventilation. In addition, all of these parameters are remotely controllable to facilitate use of the device in hostile or remote environments. The exhaled breath is also maintained at body temperature and a high humidity. The simulation is accurate to the extent of having a variable functional residual capacity independent of other parameters.

Bartlett, R. G.↗

Rare-gas effects on metabolism and inert gas narcosis

The detailed examination is reported of the theory that narcosis results from expansion of the cell membrane under high partial pressures. The research is partially based on the hypothesis that, like oxygen toxicity, the mechanism of metabolic effects of rare gases may be similar at both low and high pressures and are simply more observable at high pressures. Using adult female goats, the parameters measured include oxygen consumption, CO2 production, respiration rate, heart rate, rectal and skin temperatures and the analysis of electroencephalograms and evoked response. Additionally, the specific activity is measured of plasma glucose subsequent to injection of glucose-UL-C-14, intravenous infusion, specific activity of expired CO2, unesterified fatty acid levels and whole blood lactate-to-pyruvate ratios. Also studied were the effects of acetylsalicylic acid, vitamin E and cationic detergents (which alleviate narcosis) upon metabolic changes induced by high pressure narcosis.

Source record↗

Isolation of carbohydrate-metabolizing, extremely halophilic bacteria.

Four previously unrecognized strains of extremely halophilic bacteria that utilize carbohydrates have been isolated. Gas production proved an unreliable index of carbohydrate metabolism; therefore, carbohydrate utilization was measured by determining acid formation and sugar disappearance during growth. By these procedures, carbohydrate utilization was readily detected. The results suggest that carbohydrate dissimilation by extremely halophilic bacteria may be more common than previously thought and that the apparent rarity of carbohydrate-metabolizing halophiles may be an artifact of the isolation procedures used.

Tomlinson, G. A.↗

Breathing metabolic simulator

A description is given of an automatic computer controlled second generation breathing metabolic simulator (BMS). The simulator is used for evaluating and testing respiratory diagnostic, monitoring, support, and resuscitation equipment. Any desired sequence of metabolic activities can be simulated on the device for up to 15 hours. The computer monitors test procedures and provides printouts of test results.

Bartlett, R. G., Jr.↗

A monkey metabolism pod for space-flight weightlessness studies.

The system described will permit quantitative physiological studies in adult monkeys, weighing from 8 to 14 kg, during future space flights. The system comprises a fiberglass pod containing a comfortable restraint couch for the animal. The pod is divided into upper and lower halves. When the monkey occupies the couch, a rubber belly-band forms a gas seal between the upper and lower portions of the animal. The upper-pod ventilating air stream is monitored for the partial pressures of oxygen, carbon dioxide, nitrogen, and water to permit continuous metabolic gas-exchange measurements for computation of metabolic energy expediture. The lower pod is lined with ashless filter paper for excreta collection.

Pace, N.↗

Evidence for metabolic activity of airborne bacteria

Aerosols of the bacterium Serratia marcescens, and of uniformly labeled C-14 glucose were produced simultaneously and mixed in tubing leading to an aerosol chamber. During a subsequent period of about 5 hrs, carbon dioxide was produced metabolically within the chamber, and labeled material incorporated within the suspended particles first increased then decreased. This constitutes the first direct evidence of microbial metabolism of bacteria suspended in the air.

Chatigny, M. A.↗

Research on inert gas narcosis and air velocity effects on metabolic performance

The effects of air velocity on metabolic performance are studied by using high forced airflow in a closed environment as a mechanism to control the concentration of volatile animal wastes. Air velocities between 100 and 200 ft/min are without significant effects on the metabolism of rats. At velocities of 200 ft/min and above, oxygen consumption and CO2 production as well as food consumption increase. In most instances, the changes are on the order of 5-10%. At the same time, the RQ for the animals increases slightly and generally correlates well with oxygen consumption and CO2 production. Experiments on the nature of inert gas narcosis show that halothane and methoxyflurane are rather potent inhibitors of the NADH:O2 oxidoreductase system in rats. These experiments suggest that the mechanism of inert gas narcosis is not mandatorily related to a membrane surface phenomenon.

Source record↗

An environmental chamber system for prolonged metabolic studies on small animals

Measurement of metabolic adaptation to marginally stressful environments requires both precise regulation of a variety of atmospheric factors for extended periods of time and the capacity to employ sensitive parameters in an undisturbed subject. This paper describes a metabolic chamber system which can simultaneously maintain groups of small animals in two completely separate closed environments having different pressures, temperatures and gas compositions for an indefinite period. Oxygen consumption, carbon dioxide production, food and water consumption and animal activity cycles can be continuously monitored and quantified 24 h per day while the animals are in an unrestrained state. Each chamber can be serviced and the animals handled, injected and sacrificed without subjecting them to barometric stress. Several unique electrical and mechanical components allow semi-automated data collection on a continuous basis for indefinite periods of time.

Jordan, J. P.↗

Mineral and nitrogen balance study - Results of metabolic observations on Skylab II 28-day orbital mission

The prediction that various stresses of flight, particularly weightlessness, would bring about significant derangements in the metabolism of the musculoskeletal system has been based on various balance-study observations of long-term immobilized or inactive bed rest. The three astronauts of Skylab II consumed a planned dietary intake of major metabolic elements in mixed foods and beverages and provided virtually complete collections of excreta for 31 days preflight, 28 days inflight, and 17 days postflight. Analyses showed that, in varying degree among the crewmen, urinary calcium increased gradually during flight in a pattern similar to that observed in bed-rest studies. Fecal calcium excretion did not change significantly, but calcium balance, owing to the urinary calcium rise, became either negative or less positive than in preflight measurement. Increased excretion and negative nitrogen and phosphorus balances inflight indicated appreciable loss of muscle tissue in all three crewmen. Significant losses also occurred inflight in potassium, sodium, and magnesium. Based on the similarity in pattern and degree between these observations of calcium, phosphorus, and nitrogen loss, musculoskeletal integrity would not be threatened in space flights of up to at least 3 months. However, if similar changes occur in the planed Skylab flights for considerably more than 28 days, concern for capable musculoskeletal function should be serious for flights of very many months' duration.

Whedon, G. D.↗

Metabolic rate control during extravehicular activity simulations and measurement techniques during actual EVAS

A description of the methods used to control and measure metabolic rate during ground simulations is given. Work levels attained at the Space Environment Simulation Laboratory are presented. The techniques and data acquired during ground simulations are described and compared with inflight procedures. Data from both the Skylab and Apollo Program were utilized and emphasis is given to the methodology, both in simulation and during flight. The basic techniques of work rate assessment are described. They include oxygen consumption, which was useful for averages over long time periods, heart rate correlations based on laboratory calibrations, and liquid cooling garment temperature changes. The relative accuracy of these methods as well as the methods of real-time monitoring at the Mission Control Center are discussed. The advantages and disadvantages of each of the metabolic measurement techniques are discussed. Particular emphasis is given to the problem of utilizing oxygen decrement for short time periods and heart rate at low work levels. A summary is given of the effectiveness of work rate control and measurements; and current plans for future EVA monitoring are discussed.

David J. Horrigan↗

Metabolic analyzer

The metabolic analyzer was designed to support experiment M171. It operates on the so-called open circuit method to measure a subject's metabolic activity in terms of oxygen consumed, carbon dioxide produced, minute volume, respiratory exchange ratio, and tidal volume or vital capacity. The system operates in either of two modes. (1) In Mode I, inhaled respiratory volumes are actually measured by a piston spirometer. (2) In Mode II, inhaled volumes are calculated from the exhaled volume and the measured inhaled and exhaled nitrogen concentrations. This second mode was the prime mode for Skylab. Following is a brief description of the various subsystems and their operation.

Lem, J. D.↗

Mineral and nitrogen metabolic studies on Skylab flights and comparison with effects of earth long-term recumbency

A metabolic study of the effects of space flight on various chemical elements, particularly those with special relevance to the musculo-skeletal system, was carried out on the nine astronauts who participated in the three Skylab flights of 28, 59 and 84 days in 1973-1974. The study required of the cooperating crewmen constant dietary intake, continuous 24-hour urine collections and total fecal collections for 21-31 days before each flight, throughout each flight and for 17-18 days post-flight. Increases in urinary calcium and negative calcium balances during space flight were generally similar to those found in previous immobilization and bedrest studies. The persistence of these alterations in calcium metabolism throughout the flights suggested that calcium losses would continue in weightlessness for a very long time. Significant losses of nitrogen and phosphorus occurred, associated with observed reduction in muscle tissue. Both mineral and muscle losses occurred despite rigorous exercise regimens in flight. It was concluded that unless protective measures can be developed, capable musculo-skeletal function is likely to be impaired in space flights, ultimately to be conducted to Mars, of 1-1/2 to 3 years duration.

Whedon, G. D.↗

The correlation of sodium and potassium metabolism with the level of energy consumption in man during adaptation to heat

The sodium and potassium metabolism was studied in a thermal chamber at 35 deg and 80 percent relative humidity in 8 men for a period of 6 days. The control group (3 subjects) were outside of the chamber at a comfortable ambient temperature. The intracellular sodium and potassium metabolism were assessed based on their content in the erythrocytes. The finding was that during adaptation to heat, a considerable amount of sodium was excreted by the body in the sweat and urine (about 1/3 of the sodium content of the human body) as compared with its intake and the amount of potassium retained in the body. Changes in the concentration of sodium and potassium may serve as indexes of the state of adaptation processes during constant exposure to heat.

Afanasyev, B. G.↗