Adenylate Energy Charge Research Articles

The relationship between microbial community vitality and ATP bioburden in bottom waters under fuel microcosms.

Adenylate energy charge (AEC) - computed from the ATP, ADP and AMP concentrations in a specimen - reflect the net physiological state of the microbial population in that specimen. Previous research has demonstrated that healthy microbial populations maintain AEC≥0.8. As populations are subjected to stresses, or - in closed systems - deplete the available nutrients, respond to the accumulation of toxic metabolites, or both, AEC decreases (often to <0.5). Aqueous-phase samples from a set of fuel-water microcosms were tested for cellular ATP (cATP) and AEC. This paper reports on the precision of the AEC test method and the relationship between cellular AEC and cATP bioburdens in the aqueous phase of fuel over aqueous-phase microcosms.

Spatial probabilistic mapping of metabolite ensembles in mass spectrometry imaging

Mass spectrometry imaging vows to enable simultaneous spatially resolved investigation of hundreds of metabolites in tissues, but it primarily relies on traditional ion images for non-data-driven metabolite visualization and analysis. The rendering and interpretation of ion images neither considers nonlinearities in the resolving power of mass spectrometers nor does it yet evaluate the statistical significance of differential spatial metabolite abundance. Here, we outline the computational framework moleculaR (https://github.com/CeMOS-Mannheim/moleculaR) that is expected to improve signal reliability by data-dependent Gaussian-weighting of ion intensities and that introduces probabilistic molecular mapping of statistically significant nonrandom patterns of relative spatial abundance of metabolites-of-interest in tissue. moleculaR also enables cross-tissue statistical comparisons and collective molecular projections of entire biomolecular ensembles followed by their spatial statistical significance evaluation on a single tissue plane. It thereby fosters the spatially resolved investigation of ion milieus, lipid remodeling pathways, or complex scores like the adenylate energy charge within the same image.

Survival of Escherichia coli after high-antibiotic stress is dependent on both the pregrown physiological state and incubation conditions.

The survival of bacterial cells exposed to antibiotics depends on the mode of action, the antibiotics concentration, and the duration of treatment. However, it also depends on the physiological state of the cells and the environmental conditions. In addition, bacterial cultures contain sub-populations that can survive high antibiotic concentrations, so-called persisters. Research on persisters is challenging due to multiple mechanisms for their formation and low fractions, down to and below one millionth of the total cell population. Here, we present an improved version of the persister assay used to enumerate the amount of persisters in a cell population. The persister assay with high antibiotic stress exposure was performed at both growth supporting and non-supporting conditions. Escherichia coli cells were pregrown to various growth stages in shake flasks and bench-top bioreactors. In addition, the physiological state of E. coli before antibiotic treatment was determined by quantitative mass spectrometry-based metabolite profiling. Survival of E. coli strongly depended on whether the persister assay medium supported growth or not. The results were also highly dependent on the type of antibiotic and pregrown physiological state of the cells. Therefore, applying the same conditions is critical for consistent and comparable results. No direct connection was observed between antibiotic efficacy to the metabolic state. This also includes the energetic state (i.e., the intracellular concentration of ATP and the adenylate energy charge), which has earlier been hypothesized to be decisive for persister formation. The study provides guides and suggestions for the design of future experimentation in the research fields of persisters and antibiotic tolerance.

Dietary Nucleotides Accelerate Intestinal Recovery after Food Deprivation in Old Rats

Previous studies in very young rats have shown that dietary nucleotides improve small intestine repair after injury or malnutrition. To investigate the potential effect of nucleotides in old rats, which have a diminished capability for intestinal repair, 17-mo-old rats were deprived of food for 5 d and then fed a nucleotide-free diet or a nucleotide-supplemented diet for 3 or 6 d. Intestinal jejunal and ileal mucosal weight, protein and DNA were evaluated as intestinal growth markers, and brush-border maltase, sucrase, lactase and aminopeptidase activities were evaluated as intestinal differentiation markers. The adenine nucleotide pool and the adenylate energy charge were also evaluated as indices of nucleotide availability. Food deprivation significantly decreased mucosal growth markers as well as differentiation markers in both jejunum and ileum. The ATP pool was also significantly depressed, but the adenylate energy charge was not significantly altered. To a certain extent, refeeding restored the losses, but in the rats that were fed the nucleotide-free diet, the restoration of the jejunum was significantly slower and the restoration of the ileum differentiation markers was incomplete compared with the rats fed the nucleotide-supplemented diet. The results suggest that dietary nucleotide intake in the elderly may accelerate the normal physiological intestinal response to refeeding after food deprivation.

Biochemical composition and adenylate energy charge shifts in longfin yellowtail (Seriola rivoliana) embryos during development under different temperatures

The longfin yellowtail Seriola rivoliana is an emerging species for aquaculture diversification worldwide and production relies on fertilized eggs from captive broodstock. Temperature is the main factor that influences the developmental process and success during fish ontogeny. However, the effects of temperature on the utilization of the main biochemical reserves and bioenergetics are scarcely investigated in fish, whereas protein, lipid and carbohydrate metabolism have critical roles in maintaining cellular energy homeostasis. In this context, we aimed to evaluate metabolic fuels (protein, lipids, triacylglicerides, carbohydrates), adenylic nucleotides and derivates (ATP, ADP, AMP, IMP), and the adenylate energy charge (AEC) during embryogenesis and in hatched larvae in S. rivoliana at different temperatures. For this purpose, fertilized eggs were incubated at six constant (20, 22, 24, 26, 28 and 30 °C) and two oscillating (21⇄29 °C) temperatures. Biochemical analyses were made at blastula, optic vesicles, neurula, prehatch and hatch periods. Results indicated that the developmental period had a major influence on the biochemical composition at any temperature regime tested during the incubation. Protein content decreased only at hatching mainly due to the loss of the chorion, total lipids tended to increase at the neurula period and variations in carbohydrates depended on the particular spawn analyzed. Triacylglicerides were a critical egg fuel during hatching. The high AEC during embryogenesis and even in hatched larvae suggested an optimal energy balance regulation. The lack of critical biochemical changes from different temperature regimes during embryo development confirmed that this species exhibits a high adaptive capacity in response to constant and fluctuating temperatures. However, the timing of hatching was the most critical period of development, where biochemical components and energy utilization significantly changed. The oscillating temperatures tested may have physiological advantages without detrimental energetic effects that will require further research on larval quality after hatching.

Energy metabolism of juvenile scallops Nodipecten subnodosus under acute increased temperature and low oxygen availability

High temperature increases energy demand in ectotherms, limiting their physiological capability to cope with hypoxic events. The present study aimed to assess the metabolic tolerance of juvenile Nodipecten subnodosus scallops to acute hyperthermia combined with moderate hypoxia. A previous study showed that juveniles exhibited a high upper temperature limit (32 °C), but the responses of juveniles to combined hyperthermia and low dissolved oxygen are unknown. Scallops were exposed to control conditions (treatment C: 22 °C, ∼7.1 mg O2 L−1 or PO2 156.9 mmHg), acute hyperthermia under normoxia (treatment T: 30 °C, ∼6.0 mg O2 L−1 or PO2 150.9 mmHg) or acute hyperthermia plus hypoxia (treatment TH: 30 °C, ∼2.5 mg O2 L−1 or PO2 62.5 mmHg) for 18 h. In T, juveniles exhibited an enhanced oxygen consumption, together with a decrease in adenylate energy charge (AEC) and arginine phosphate (ArgP), and with no changes in metabolic enzyme activity in the muscle. In TH, scallops maintained similar AEC and ArgP levels in muscle as those observed in T treatment. This response occurred along with the accumulation of inosine monophosphate and hypoxanthine. Besides, reduced citrate synthase and pyruvate kinase activities, enhanced hexokinase activity, and a higher octopine dehydrogenase/lactate dehydrogenase ratio in the mantle indicated the onset of anaerobiosis in TH. These responses indicate that juvenile scallops showed tissue-specific compensatory responses regarding their energy balance under moderate hypoxia at high temperatures. Our results give an insight into the tolerance limit of this species to combined hyperthermia and hypoxia in its northern limit of distribution.

Effect of super-chilling storage on maintenance of quality and freshness of the Pacific oyster (Crassostrea gigas)

AbstractThe quality changes of shelled Pacific oysters (Crassostrea gigas) were examined in relation to the effects of superchilling storage at −1 °C for 28 d by measuring changes in biochemical properties (microbial analysis, adenosine triphosphate (ATP)-related compounds, pH, free amino acids) and sensory evaluations in this study. The results indicated that microorganism growth was significantly inhibited during superchilling storage. Adenosine diphosphate (ADP) and adenosine monophosphate (AMP) accumulated while ATP rapidly decreased in the adductor muscle. ATP and ADP were the primary components in the other 3 tissues including mantle, gill, and body trunk of oysters, and they remained ­relatively stable over time. The pH and adenylate energy charge in the adductor muscle could be utilized as freshness indicators for shelled oysters. However, there were no significant differences (P&amp;gt;0.05) among the free amino acids during whole storage. According to the sensory evaluations, oysters could be alive and tolerated up to 21 d at −1 °C storage. The study demonstrated that superchilling storage at −1 °C could better maintain the eating quality of shelled oysters and the shelf life was extended to 21 d.

Changes in Metabolic Rate, Excretion, Energy Reserves, and Starvation Response of Diploid and Triploid Salvelinus Fontinalis and Diploid O. Mykiss after LongTerm Exposure to Elevated Temperature

Background: In fish culture metabolic rate (oxygen consumption), and excretion of particular and dissolved waste products are important parameters limiting the production capacity and influencing the environment. Data on their changes in response to elevated temperature are important for future management of fish farms. Therefore, these parameters were investigated in diploid (2n) and triploid (3n) Salvelinus fontinalis and 2n Oncorhynchus mykiss exposed to 20°C for 32 d in comparison to fish acclimated to 9 °C. Additionally energy reserves were investigated as important indicators for fish performance. Results: At 20°C routine metabolic rate (RMR) of 2n S. fontinalis decreased with increasing exposure time and after 32 d it did not differ from 9 °C. Also, in O. mykiss RMR decreased with exposure time but remained 60 % higher than at 9°C. In 3n S. fontinalis exposed to 20°C RMR was constantly increased for 120%. For all species/ploidy levels faeces dry weight and phosphorus concentration and quantities of excreted dissolved nitrogen and phosphate did not differ between 9°C and 20°C. Lipid concentration of faeces was decreased at 20°C. With exception of O. mykiss also faeces protein was decreased at 20°C. In all species/ploidy levels liver glycogen and visceral fat was decreased at 20°C, while liver triglycerides and adenylate energy charge were not affected. In 2n and 3n S. Salvelinus temperature related responses to 5-d starvation were investigated. In 2n S. fontinalis RMR decreased in starving fish in comparison to fed ones at 9°C and increased at 20°C. RMR of 3n S. fontinalis did not change. For both ploidy levels, dissolved excretion was decreased due to starvation at 9°C and 20°C. Visceral fat decreased due to starving at 9°C, liver glycogen and triglycerides at 20°C. Conclusion: In Salmonidae species and ploidy dependent types of metabolic thermal compensation occurred after exposure to 20°C. Effects on waste products were minor for all investigated species and ploidy levels. Exposure to 20°C represented an energetic stress situation as the visceral fat and liver glycogen depots were depleted. Consequently, at elevated temperature starving can result in a critical energetic stress situation which might lead to complete exhaustion of energy resources.

Nucleotides and free amino acids in sea urchin Paracentrotus lividus gonads: Contributions for freshness and overall taste

This study aims to report the chemical changes, specifically the nucleotides and free amino acids (FAAs) during refrigerated storage of live sea urchin Paracentrotus lividus (7 °C) and packed gonads (2 °C) and respective contributions to overall taste. Results showed that the adenylate energy charge (AEC) is an adequate indicator for live sea urchin through storage, while K-value does not distinguish freshness. Changes of both indexes were not clear in packed gonads. The FAAs profile were related with maturation stage where amino acids associated to sweet (Gly, Ala), umami (Glu) and bitter (Arg, Lys, His, Val) had significant contribution to overall taste. In general, the storage did not significantly affect the FAAs content nor its contribution to overall taste in contrast to nucleotides that seemed to induce potential changes after day 5. The findings provide insights that will complement a full study on quality index scheme development of the products.

Temperature limits for storage of extended boar semen from the perspective of the sperm's energy status.

The optimum storage temperature for liquid-preserved boar semen has been empirically determined to be between 15 and 20°C. Lower temperatures provide an advantage to inhibit bacterial growth, but are regarded as critical due to the high sensitivity of boar spermatozoa to chilling injury. Higher storage temperatures are supposed to induce energy deficiency due to an insufficient depression of metabolic cell activity. However, experimental evidence for alterations of the sperm's energy status in relation to storage temperature and duration is missing. Therefore, we aimed to revisit the upper and lower storage temperature limits for liquid-preserved boar semen from the perspective of the sperm's energy metabolism. Ejaculates (n = 7 boars) were cooled down in Beltsville Thawing Solution (BTS) to 25, 17, 10, or 5°C and stored for up to 120 h. ATP and adenylate energy charge (EC) levels were assessed at storage temperature (24, 72, and 120 h storage) and after subsequent re-warming (38°C). Sperm quality and energy status remained at a stable level in samples stored at 25 and 17°C. Chilling to and storage at 10 or 5°C in BTS provoked cold shock in a subset of sperm as shown by a loss in viability and motility (P < 0.05), which was accompanied by a significant release of adenine nucleotides into the semen extender. Prolonged storage for 120 h resulted in significantly lower mean ATP concentrations in viable spermatozoa at 5 or 10°C compared to 17°C (P < 0.05). Cluster analysis revealed that the main sperm subpopulation, i.e., sperm with moderate speed and linearity, decreased from 50 to 30% (P < 0.05) in favor of slow-moving spermatozoa (5°C) or spermatozoa with a hyperactivation-like motility pattern (10°C). The results point to a sublethal imbalance in available ATP in a subset of the surviving sperm population, rather than a general decrease in available ATP in all spermatozoa. In conclusion, storing diluted boar semen at a stable temperature between 17 and 25°C is a safe procedure concerning the spermatozoa's energy status. Future concepts for hypothermic boar semen preservation below 17°C require measures which ameliorate the imbalanced energy status in viable spermatozoa.

Effect of acute hypoxia on the brain energy metabolism of the scorpionfish Scorpaena porcus Linnaeus, 1758: the pattern of oxidoreductase activity and adenylate system.

The activity of oxidoreductases, malate dehydrogenase and lactate dehydrogenase (MDH, 1.1.1.37; LDH, 1.1.1.27), as well as parameters of adenylate system-[ATP], [ADP], [AMP], total adenylate pool (AP), and adenylate energy charge (AEC) in medulla oblongata (MB) and forebrain, midbrain, and diencephalon (FDMB)-were studied in the scorpionfish under acute hypoxia (0.9-1.2mgO2·L-1, 90min). A higher MDH activity level was observed in MB and FDMB, as compared to LDH (p < 0.05). At the same time, MB showed a higher adenylate content and increased AP (p < 0.05). AEC did not exceed ~ 0.7 (vs. the maximum of this index ~ 0.9-1.0) in the brain of the scorpionfish indicating adaptation of the tissue energy status to hypoxia. A rapid decrease in MDH activity (p < 0.05) was observed in MB under acute hypoxia. These changes were accompanied by insignificant LDH activation. A pronounced LDH activation (p < 0.05), a decrease in MDH activity, and the highest AP raise (p < 0.05) were observed in FDMB, suggesting activation of glycolysis and simultaneous decrease in the rate of ATP consumption. MB and FDMB demonstrated the ability to a relative retention of AEC during hypoxia. The unidirectional metabolic adaptation was based on the intensification of glycolysis, a decrease of ATP consumption, and a subsequent increase in adenylate concentration that allowed the scorpionfish brain structures to maintain the energy status under acute hypoxia.

AMPK regulates behavior and physiological plasticity of Haliotis discus hannai under different spectral compositions

In natural environments, the spectral composition of incident light is often subject to drastic changes due to the abundance of suspended particles, floating animals, and plants in coastal waters. In this study, after four months of culturing under blue light (NB), orange light (NY), dark environment (ND), and natural light (NN), the shell length and weight-specific growth rate in Pacific abalone, Haliotis discus hannai, were ranked in the following order: NY > NN > ND > NB. To understand the growth differences in abalone under these different light environments, we first performed 24-h video monitoring and found that the cumulative movement distance and duration were lowest in group NB, whereas the cumulative movement distance and duration were significantly higher in group ND than in any other group (P < 0.05). In group NB, the time spent hidden underneath the attachment substrate accounted for 81% of the resting time, but this ratio was lowest in group ND, at only 37% (P < 0.05). Next, LC-MS metabolomics identified 201 and 105 metabolites in NB vs. NN, ND vs. NN, and NY vs. NN under the positive and negative ion modes, respectively. According to the fold changes and annotations for differential metabolites in the KEGG enrichment pathways, adenosine, NAD+, cGMP, and arachidonic acid were used as differential metabolism markers, and the AMPK signaling pathway was enriched in every comparison group, and thus investigated further. The gene sequences of three subtypes of AMPK were obtained by cloning and we found that the expression levels of AMPKα and AMPKγ, and the AMP content were significantly higher in group NB than in any other group (P < 0.05). In addition, the ATP contents and adenylate energy charge values were ranked in the following order: NY > NN > ND > NB. According to in situ hybridization analysis, the three subtype genes were widely expressed in the hepatopancreas. Finally, the contents of many lipid metabolites differed significantly among groups and the expression levels of the triglyceride hydrolysis-related gene hormone sensitive lipase and fatty acid oxidation-related gene carnitine palmitoyltransferase 1 were higher in groups ND and NB than in groups NN and NY according to fluorescence quantification PCR (P < 0.05). The expression levels of fatty acid synthase and acetyl-CoA carboxylase were significantly lower in groups ND and NB than in groups NN and NY (P < 0.05). These findings indicated that differences in the spectral composition of incident light could reshape the behavior and physiological metabolism in abalone by influencing the “energy switch” AMPK, thereby providing some insights into the mechanisms that allow nocturnal marine organisms to adapt to different lighting environments.

Melatonin Alleviates Chilling Injury Symptom Development in Mango Fruit by Maintaining Intracellular Energy and Cell Wall and Membrane Stability.

The efficacy of the signaling molecule melatonin for alleviating chilling injury (CI) in mango (Mangifera indica L.) fruit was studied to investigate the potential role of membrane integrity, energy charge, and ripening-related changes in the development of CI, and its management by melatonin. ‘Langra’ and ‘Gulab Jamun’ cultivar mango fruit was immersed in 100 μM of melatonin before storage for 28 days at 5°C with weekly transfers to shelf life at 25°C. CI symptom development was associated with compositional and enzymatic aspects of textural changes, cell membrane deterioration, and chemical energy status. Melatonin-treated ‘Langra’ fruit exhibited very low CI (5 vs. 21%) while ‘Gulab Jamun’ fruit exhibited higher CI (36 vs. 38%) during 28 days of storage at 5 ± 1°C. Higher chilling tolerance in melatonin-treated ‘Langra’ was associated with lower softening, ascribed to lower cell wall degrading exo- and endo-polygalacturonase, pectinesterase, and endo-1,4-β-D-glucanase. In addition, lower membrane deteriorating-phospholipase D and lipoxygenase activity in melatonin-treated ‘Langra’ corresponded to lower palmitic and stearic acids and higher oleic, linoleic, and linolenic acids accumulation, thus, higher unsaturated/saturated fatty acids ratio. Additionally, there was a higher intracellular energy supply with melatonin, represented by a higher adenylate energy charge (AEC) arising from higher ATP and ADP and lower AMP accumulation, related to higher H+-ATPase, Ca2+-ATPase, succinate dehydrogenase, and cytochrome c oxidase activities. This study for the first time provides evidence, suggesting that melatonin alleviation of CI is related to the preservation of membrane integrity, thereby protecting the intracellular energy supply, and preserving cell wall integrity via impeding cell wall degrading enzyme activities.

Multi‑omics analysis of right ventricles in rat models of pulmonary arterial hypertension: Consideration of mitochondrial biogenesis by chrysin.

In pulmonary arterial hypertension (PAH), right ventricular failure is accompanied by metabolic alterations in cardiomyocytes, which may be due to mitochondrial dysfunction and decreased energy production. Chrysin (CH) is a phytochemical with pharmacological activity that is involved in the regulation of mitochondrial biogenesis. The present study investigated the role of CH in the right ventricle (RV) by analyzing the cardiac transcriptome and metabolome of a SU5416(a vascular endothelial growth factor receptor blocker,/hypoxia (Su/Hx) rat model of PAH. RNA-sequencing of the RV transcriptome between Su/Hx, Su/Hx with CH (Su/Hx + CH) and control groups, extracellular matrix (ECM) organization and ECM-receptor interaction-associated genes were upregulated in the RV of Su/Hx but not Su/Hx + CH rats. Furthermore, expression of mitochondrial function-, energy production-, oxidative phosphorylation- and tricarboxylic acid (TCA) cycle-associated genes was decreased in the RV of Su/Hx rats; this was reverse by CH. Metabolomic profiling analysis of Su/Hx and Su/Hx + CH rats showed no significant changes in glycolysis, TCA cycle, glutathione, NADH or NADPH. By contrast, in the RV of Su/Hx rats, decreased adenylate energy charge was partially reversed by CH administration, suggesting that CH was involved in the improvement of mitochondrial biogenesis. Reverse transcription-quantitative PCR analysis revealed that expression of peroxisome proliferator-activated receptor γ, a master regulator of fatty acid metabolism and mitochondrial biogenesis, was increased in the RV of Su/Hx + CH rats. CH ameliorated cardiac abnormality, including cardiac fibrosis, RV hypertrophy and PH. The present study suggested that CH altered patterns of gene expression and levels of mitochondrial metabolites in cardiomyocytes, thus improving RV dysfunction in a Su/Hx PAH rat model.

Combined Targeted and Untargeted Profiling of HeLa Cells Deficient in Purine De Novo Synthesis.

Three genetically determined enzyme defects of purine de novo synthesis (PDNS) have been identified so far in humans: adenylosuccinate lyase (ADSL) deficiency, 5-amino-4-imidazole carboxamide-ribosiduria (AICA-ribosiduria), and deficiency in bifunctional enzyme phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazolesuccinocarboxamide synthase (PAICS). Clinical signs of these defects are mainly neurological, such as seizures, psychomotor retardation, epilepsy, autistic features, etc. This work aims to describe the metabolic changes of CRISPR-Cas9 genome-edited HeLa cells deficient in the individual steps of PDNS to better understand known and potential defects of the pathway in humans. High-performance liquid chromatography coupled with mass spectrometry was used for both targeted and untargeted metabolomic analyses. The statistically significant features from the untargeted study were identified by fragmentation analysis. Data from the targeted analysis were processed in Cytoscape software to visualize the most affected metabolic pathways. Statistical significance of PDNS intermediates preceding deficient enzymes was the highest (p-values 10 × 10−7–10 × 10−15) in comparison with the metabolites from other pathways (p-values of up to 10 × 10−7). Disturbed PDNS resulted in an altered pool of adenine and guanine nucleotides. However, the adenylate energy charge was not different from controls. Different profiles of acylcarnitines observed among deficient cell lines might be associated with a specific enzyme deficiency rather than global changes related to the PDNS pathway. Changes detected in one-carbon metabolism might reduce the methylation activity of the deficient cells, thus affecting the modification state of DNA, RNA, and proteins.

Creatine transport and creatine kinase activity is required for CD8+ Tcell immunity.

The factors that promote Tcell expansion are not fully known. Creatine is an abundant circulating metabolite that has recently been implicated in Tcell function; however, its cell-autonomous role in immune-cell function is unknown. Here, we show that creatine supports cell-intrinsic CD8+ Tcell homeostasis. We further identify creatine kinase B (CKB) as the creatine kinase isoenzyme that supports these Tcell properties. Loss of the creatine transporter (Slc6a8) or Ckb results in compromised CD8+ Tcell expansion in response to infection without influencing adenylate energy charge. Rather, loss of Slc6a8 or Ckb disrupts naive Tcell homeostasis and weakens TCR-mediated activation of mechanistic target of rapamycin complex 1 (mTORC1) signaling required for CD8+ Tcell expansion. These data demonstrate a cell-intrinsic role for creatine transport and creatine transphosphorylation, independent of their effects on global cellular energy charge, in supporting CD8+ Tcell homeostasis and effector function.

Assessing the Energy Status of Low Numbers of Mammalian Cells by Capillary Electrophoresis-Mass Spectrometry.

Capillary electrophoresis-mass spectrometry (CE-MS) employing a sheathless porous tip interface has become a strong analytical tool for the efficient profiling of highly polar and charged metabolites in volume/material-restricted biological samples. As more and more metabolomics studies are (intrinsically) dealing with low numbers of mammalian cells, it would be important to use an additional performance metric to effectively evaluate the sampling and sample preparation procedure, in particular quenching. An established parameter to assess the sampling and sample preparation quality when working with cell cultures is the adenylate energy charge (AEC), which represents an index of the energy state of a cell. In this protocol, a CE-MS strategy is proposed for the reliable determination of the adenylate energy charge (AEC) in metabolomics studies dealing with low numbers of mammalian cells.

Impact of transportation and rehydration strategies on the physiological responses of clams (Ruditapes philippinarum)

With the prolongation of the post-catch circulation time, the soft tissue of live clam could gradually decline in its quality. This study investigated the quality changes of clams (Ruditapes philippinarum) during the cold chain transportation, and then focused on the rehydration processes, which included gradient heating rehydration (GR) and sudden heating rehydration (SR). Results showed that a higher survival rate of clams was obtained in the GR stage (95%) clams than that in the SR stage (76%). In addition, both glycogen content and adenylate energy charge (A.E.C) value were also higher in the GR stage (20.49 ± 1.05 mg/g, 70.77 ± 1.76%) than that in the SR stage (15.47 ± 1.83 mg/g, 61.63 ± 1.74%) clams. Conversely, the clams in the GR stage showed lower lactic acid content (0.167 ± 0.003 mmol/g) than that in the SR stage (0.181 ± 0.004 mmol/g). Therefore, during the rehydration processes, the GR was a better approach to preserve the quality of clams when compared with the SR. This work could provide reference to preserve the quality of clams and reduce the economic loss of clams during the transportation and rehydration process.

Physiological and Molecular Responses in the Gill of the Swimming Crab Portunus trituberculatus During Long-Term Ammonia Stress

Ammonia is a common environmental stressor encountered during aquaculture, and is a significant concern due to its adverse biological effects on vertebrate and invertebrate including crustaceans. However, little information is available on physiological and molecular responses in crustaceans under long-term ammonia exposure, which often occurs in aquaculture practices. Here, we investigated temporal physiological and molecular responses in the gills, the main ammonia excretion organ, of the swimming crab Portunus trituberculatus following long-term (4 weeks) exposure to three different ammonia nitrogen concentrations (2, 4, and 8 mg l–1), in comparison to seawater (ammonia nitrogen below 0.03 mg l–1). The results revealed that after ammonia stress, the ammonia excretion and detoxification pathways were initially up-regulated. These processes appear compromised as the exposure duration extended, leading to accumulation of hemolymph ammonia, which coincided with the reduction of adenosine 5′-triphosphate (ATP) and adenylate energy charge (AEC). Considering that ammonia excretion and detoxification are highly energy-consuming, the depression of these pathways are, at least partly, associated with disruption of energy homeostasis in gills after prolonged ammonia exposure. Furthermore, our results indicated that long-term ammonia exposure can impair the antioxidant defense and result in increased lipid peroxidation, as well as induce endoplasmic reticulum stress, which in turn lead to apoptosis through p53-bax pathway in gills of the swimming crab. The findings of the present study further our understanding of adverse effects and underlying mechanisms of long-term ammonia in decapods, and provide valuable information for aquaculture management of P. trituberculatus.

Temperature effects on the nutritional quality in Pacific oysters (Crassostrea gigas) during ultraviolet depuration

Oysters are mainly consumed in the raw form, so it is important to get rid of bacteria and other harmful substances. Ultraviolet (UV) sterilization depuration is a commonly used method and does not produce chemical residues or act directly on shellfish, resulting in minimal adverse effects on flavor. This study simulated the industrial depuration process using UV sterilization to depurate Pacific oysters (Crassostrea gigas). The effects of different temperatures (15, 20, and 25 °C) on the quality and taste components of C. gigas were investigated by measuring changes in physiological and biochemical indexes in C. gigas tissue samples. At the end of depuration, no oyster mortality occurred, but it was up to 55% at 25 °C at 84 h. Glycogen content decreased the most at 25 °C at 48 h. The fatty acid content was higher at 20 and 25 °C. Succinic acid content decreased significantly and was higher at 20 and 25 °C at 48 h with no significant difference. Total free amino acid (FAA) content was significantly higher at 20 °C, however, there were no significant differences in nucleotide content at any temperature at 48 h. Adenylate energy charge (AEC) values decreased, with higher values at 15 and 25 °C, and equivalent umami concentration (EUC) values increased, with higher values at 20 and 25 °C. Considering the changes in flavor substances and mortality rate, 20°C is the appropriate temperature for UV sterilization depuration of C. gigas to produce better edible quality. © 2021 Society of Chemical Industry.