Net Synthesis Research Articles

Path toward "Net Zero Organic Synthesis".

Researchers over the past ∼200 years have accomplished the synthesis of simple to very complex molecules; however, the concept of ideal synthesis has still not reached maturity. Of late, the "Net Zero" concept has captured the imagination of many fields of technology, in tune with Ideal Synthesis. The current Viewpoint covers the principles of ideal synthesis being discussed in the literature and how one could take up the synthesis of organic molecules considering the Net Zero concept to make this central science well-accepted by critics of this important field.

The anabolic response to protein ingestion during recovery from exercise has no upper limit in magnitude and duration in vivo in humans

The belief that the anabolic response to feeding during postexercise recovery is transient and has an upper limit and that excess amino acids are being oxidized lacks scientific proof. Using a comprehensive quadruple isotope tracer feeding-infusion approach, we show that the ingestion of 100g protein results in a greater and more prolonged (>12 h) anabolic response when compared to the ingestion of 25g protein. We demonstrate a dose-response increase in dietary-protein-derived plasma amino acid availability and subsequent incorporation into muscle protein. Ingestion of a large bolus of protein further increases whole-body protein net balance, mixed-muscle, myofibrillar, muscle connective, and plasma protein synthesis rates. Protein ingestion has a negligible impact on whole-body protein breakdown rates or amino acid oxidation rates. These findings demonstrate that the magnitude and duration of the anabolic response to protein ingestion is not restricted and has previously been underestimated invivo in humans.

Arginine and Lysine Supplementation Potentiates the Beneficial β-Hydroxy ß-Methyl Butyrate (HMB) Effects on Skeletal Muscle in a Rat Model of Diabetes.

Skeletal muscle is the key tissue for maintaining protein and glucose homeostasis, having a profound impact on the development of diabetes. Diabetes causes deleterious changes in terms of loss of muscle mass, which will contribute to reduced glucose uptake and therefore progression of the disease. Nutritional approaches in diabetes have been directed to increase muscle glucose uptake, and improving protein turnover has been at least partially an oversight. In muscle, β-hydroxy β-methyl butyrate (HMB) promotes net protein synthesis, while arginine and lysine increase glucose uptake, albeit their effects on promoting protein synthesis are limited. This study evaluates if the combination of HMB, lysine, and arginine could prevent the loss of muscle mass and function, reducing the progression of diabetes. Therefore, the combination of these ingredients was tested in vitro and in vivo. In muscle cell cultures, the supplementation enhances glucose uptake and net protein synthesis due to an increase in the amount of GLUT4 transporter and stimulation of the insulin-dependent signaling pathway involving IRS-1 and Akt. In vivo, using a rat model of diabetes, the supplementation increases lean body mass and insulin sensitivity and decreases blood glucose and serum glycosylated hemoglobin. In treated animals, an increase in GLUT4, creatine kinase, and Akt phosphorylation was detected, demonstrating the synergic effects of the three ingredients. Our findings showed that nutritional formulations based on the combination of HMB, lysine, and arginine are effective, not only to control blood glucose levels but also to prevent skeletal muscle atrophy associated with the progression of diabetes.

Whole Genome Expression Profiling of Semitendinosus Tendons from Children with Diplegic and Tetraplegic Cerebral Palsy.

Cerebral palsy (CP) is the most common movement disorder in children, with a prevalence ranging from 1.5 to 4 per 1000 live births. CP is caused by a non-progressive lesion of the developing brain, leading to progressive alterations of the musculoskeletal system, including spasticity, often leading to the development of fixed contractures, necessitating tendon lengthening surgery. Total RNA-sequencing analysis was performed on semitendinosus tendons from diplegic and tetraplegic CP patients subjected to tendon lengthening surgery compared to control patients undergoing anterior cruciate ligament reconstructive surgery. Tetraplegic CP patients showed increased expression of genes implicated in collagen synthesis and extracellular matrix (ECM) turnover, while only minor changes were observed in diplegic CP patients. In addition, tendons from tetraplegic CP patients showed an enrichment for upregulated genes involved in vesicle-mediated transport and downregulated genes involved in cytokine and apoptotic signaling. Overall, our results indicate increased ECM turnover with increased net synthesis of collagen in tetraplegic CP patients without activation of inflammatory and apoptotic pathways, similar to observations in athletes where ECM remodeling results in increased tendon stiffness and tensile strength. Nevertheless, the resulting increased tendon stiffness is an important issue in clinical practice, where surgery is often required to restore joint mobility.

Seasonal Dynamics of Protein Synthesis in the Tissues Of <i>Channa punctatus</i> [Bloch], an Indian Air-Breathing Teleost

The rates of initial synthesis and maximal net synthesis of total protein at the prevailing water-temperatures were determined by measuring the 14 levels of incorporation at 1h and 6h after administration of C-leucine respectively into total protein of liver, gill, brain, white muscle and red muscle of Chana punctatus in different months in an annual cycle. A clear biphasic pattern of seasonal variation of protein synthesis ( particularly of the net synthesis) and break-down i s evident in liver, gill and brain. This is much less discernible in muscles (white and red) . Five distinct periods of the seasonal dynamics of protein synthesis and degradation have been described as "the rising phase-I" (January-April/May-June), "the declining phase" (April/May-June-July), "the rising phase-II"(July-August) , "the maintenance phase-I" (August-October/November),"the maintenance phase-II" (October/November-December/January), and their significance discussed.

Nitrogen Balance at the Recommended Dietary Allowance for Protein in Minimally Active Male Vegans.

Vegan diets have gained popularity in recent years for reasons including health benefits and concerns for animal welfare. Although these diets are considered to be nutritionally adequate, questions remain over whether the current protein recommendation (0.8 g/kg/d) is sufficient. Protein status is determined through a nitrogen balance analysis when the protein content of the diet is known. A negative balance indicates a catabolic state, and a positive nitrogen balance indicates an anabolic state. In healthy adults, nitrogen equilibrium is the expectation reflecting the net synthesis and breakdown of proteins. Currently, there are no known studies measuring nitrogen balance in strict vegan men fed the protein requirement. Eighteen minimally active vegan men received a 5-day eucaloric diet (protein content: 0.8 g/kg/d). On day five, 24 h urine was collected for nitrogen analysis. Both the mean absolute nitrogen balance (-1.38 ± 1.22 g/d) and the mean relative nitrogen balance (-18.60 ± 16.96 mg/kg/d) were significantly lower than zero (equilibrium) (p < 0.001). There were no correlations seen between nitrogen balance and age, years as vegan, or fat-free mass. Consuming 0.8 g/kg/d of protein is not adequate to produce nitrogen balance in men adhering to typical strict vegan diets for at least one year.

Some Basic Techniques Allowing Petri Net Synthesis: Complexity and Algorithmic Issues

In Petri net synthesis we ask whether a given transition system A can be implemented by a Petri net N. Depending on the level of accuracy, there are three ways how N can implement A: an embedding, the least accurate implementation, preserves only the diversity of states of A; a language simulation already preserves exactly the language of A; a realization, the most accurate implementation, realizes the behavior of A exactly. However, whatever the sought implementation, a corresponding net does not always exist. In this case, it was suggested to modify the input behavior – of course as little as possible. Since transition systems consist of states, events and edges, these components appear as a natural choice for modifications. In this paper we show that the task of converting an unimplementable transition system into an implementable one by removing as few states or events or edges as possible is NP-complete –regardless of what type of implementation we are aiming for; we also show that the corresponding parameterized problems are W[2]-hard, where the number of removed components is considered as the parameter; finally, we show there is no c-approximation algorithm (with a polynomial running time) for neither of these problems, for every constant c ≥ 1.

Synthesis of Pure and Impure Petri Nets with Restricted Place-environments: Complexity Issues

Petri net synthesis consists in deciding for a given transition system A whether there exists a Petri net N whose reachability graph is isomorphic to A. Several works examined the synthesis of Petri net subclasses that restrict, for every place p of the net, the cardinality of its preset or of its postset or both in advance by small natural numbers ϱ and κ, respectively, such as for example (weighted) marked graphs, (weighted) T-systems and choice-free nets. In this paper, we study the synthesis aiming at Petri nets which have such restricted place environments, from the viewpoint of classical and parameterized complexity: We first show that, for any fixed natural numbers ϱ and κ, deciding whether for a given transition system A there is a Petri net N such that (1) its reachability graph is isomorphic to A and (2) for every place p of N the preset of p has at most ϱ and the postset of p has at most κ elements is doable in polynomial time. Secondly, we introduce a modified version of the problem, namely ENVIRONMENT RESTRICTED SYNTHESIS (ERS, for short), where ϱ and κ are part of the input, and show that ERS is NPcomplete, regardless whether the sought net is impure or pure. In case of the impure nets, our methods also imply that ERS parameterized by ϱ + κ is W[2]-hard.

Measuring muscle protein synthesis in humans and the influence of nutritional state.

In 1982 and 2011, Clinical Science published papers that used infusion of stable isotope-labeled amino acids to assess skeletal muscle protein synthesis in the fasted and fed state and before and after a period of increased intake of omega-3 fatty acids, respectively; both of these papers have been highly cited. An overview of the study designs, key findings and novel features, and a consideration of the lasting impact of these two papers is presented. The earlier paper introduced stable isotope tracer approaches in humans that showed consuming a meal will increase whole body oxidation, synthesis, and breakdown of protein, but that protein synthesis is greater than breakdown resulting in net accumulation of protein. The paper also demonstrated that consuming a meal promotes net protein synthesis in skeletal muscle. The later paper introduced the concept that omega-3 polyunsaturated fatty acids are able to improve anabolism by reporting that 8 weeks consumption of high-dose omega-3 fatty acids by healthy young and middle-aged adults increased skeletal muscle protein synthesis during a hyperaminoacidemic-hyperinsulinemic clamp compared with what was seen during the clamp at study entry. Omega-3 fatty acids also increased the phosphorylation of important signaling proteins in muscle, including mammalian target of rapamycin, p70s6k, and Akt, during the clamp. These two papers remain relevant because they offer experimental approaches to study human (patho)physiology in different contexts, they present novel insights into the impact of nutritional state (feeding) and specific nutrients (omega-3 fatty acids) on muscle protein synthesis, and they suggest ways to explore the potential of interventions to help prevent and reverse the age-, disease-, and disuse-associated decline in muscle mass.

Branched-chain amino acids regulate intracellular protein turnover in porcine mammary epithelial cells.

Dietary supplementation with branched-chain amino acids (BCAAs) to lactating sows has been reported to enhance their milk production, but the underlying mechanisms remain largely unknown. This study was conducted with porcine mammary epithelial cells (PMECs) to test the hypothesis that individual BCAAs or their mixture stimulates protein synthesis and inhibit proteolysis in PMECs. Cells were cultured at 37°C in customizedDulbecco's modified Eagle medium containing 5mmol/L D-glucose, 1mmol/L L-phenylalanine, L-[ring-2,4-3H]phenylalanine, 0.1 (control), 0.25, 0.5, 1, or 2mmol/L L-leucine, L-isoleucine or L-valine or an equimolar mixture of the three BCAAs. The culture medium also contained physiological concentrations of other amino acids found in the plasma of lactating sows. Proliferation, protein synthesis, proteolysis, β-casein production, the mechanistic target of rapamycin (mTOR) signaling, and the ubiquitin-proteasome pathway were determined for PMECs. Cell proliferation and abundances of phosphorylated mTOR, eukaryotic translation initiation factor 4E-binding protein 1, and ribosomal protein S6 kinase β-1 proteins increased (P < 0.05), but abundances of ubiquitinated protein and 20S proteasome decreased (P < 0.05) when extracellular concentrations of L-leucine, L-isoleucine, L-valine, or an equimolar mixture of BCAAs were increased from 0.1 to 2mmol/L. Compared with the control, 0.25, 0.5, 1 or 2mmol/L BCAAs enhanced (P < 0.01) protein (including β-casein) synthesis, while decreasing (P < 0.05) proteolysis in PMECs in a dose-dependent manner. Collectively, our results indicate that physiological concentrations of BCAAs regulate protein turnover in mammary epithelial cells to favor net protein synthesis through stimulating the mTOR signaling pathway and inhibiting the ubiquitin-proteasome pathway.

Amino Acids Supplied through the Autophagy/Endocytosis Pathway Promote Starch Synthesis in Physcomitrella Protonemal Cells.

The physiological implications of autophagy in plant cells have not been fully elucidated. Therefore, we investigated the consequences of autophagy in the moss Physcomitrella by measuring biochemical parameters (fresh and dry weights; starch, amino acid, carbohydrate, and NH3 content) in wild-type (WT) and autophagy-deficient atg5 Physcomitrella cells. We found higher starch levels and a higher net starch synthesis rate in WT cells than in atg5 cells cultured in a glucose-containing culture medium, whereas net starch degradation was similar in the two strains cultured in a glucose-deficient culture medium. Additionally, the treatment of cells with the autophagy inhibitor 3-methyladenine suppressed starch synthesis. Loading bovine serum albumin into atg5 cells through endocytosis, i.e., supplying proteins to vacuoles in the same way as through autophagy, accelerated starch synthesis, whereas loading glutamine through the plasma membrane had no such effect, suggesting that Physcomitrella cells distinguish between different amino acid supply pathways. After net starch synthesis, NH3 levels increased in WT cells, although the change in total amino acid content did not differ between WT and atg5 cells, indicating that autophagy-produced amino acids are oxidized rapidly. We conclude that autophagy promotes starch synthesis in Physcomitrella by supplying the energy obtained by oxidizing autophagy-produced amino acids.

A Novel Intron-Encoded Neuropilin-1 Isoform in Pancreatic Islets Associated With Very Young Age of Onset of Type 1 Diabetes.

Net synthesis of pancreatic β-cells peaks before 2 years of life. β-Cell mass is set within the first 5 years of life. In-frame translational readthrough of the NRP1 gene exon 9 into intron 9 generates a truncated neuropilin-1 protein lacking downstream sequence necessary for binding VEGF that stimulates β-cell replication. VEGF is critical for developing but not adult islet neogenesis. Herein we show that cells in human pancreatic islets containing the full-length neuropilin-1 possess insulin but cells that contain the truncated neuropilin-1 are devoid of insulin. Decreased insulin cells increases susceptibility to onset of type 1 diabetes at a younger age. We also show that the frequency of a genetic marker in NRP1 intron 9 is higher among patients with onset of type 1 diabetes before age 4 years (31.8%), including those with onset at 0.67-2.00 and 2-4 years, compared with that in patients with onset at 4-8 years, at 8-12 years, and after 16 years (16.1%) with frequency equal to that in subjects without diabetes (16.0%). Decreased insulin cells plus the genetic data are consistent with a low effect mechanism that alters the onset of type 1 diabetes to a very young age in some patients, thus supporting the endotype concept that type 1 diabetes is a heterogeneous disease.

Continuous oral stable isotope ingestion to measure whole-body protein turnover.

Protein kinetic responses to nutrition and exercise interventions are commonly evaluated using a primed-constant infusion of stable isotope tracers. While this methodology is state-of-the-art, the required preparation at a certified pharmacy makes the utilization of isotope infusion both expensive and logistically cumbersome. Oral tracer ingestion has been used to quantify 24-hwhole-body protein status; however, this does not permit examination of acute interventional effects. Ingestion of a priming bolus, followed by continuous ingestion of stable isotope tracer in a 'sip feeding' fashion may provide a more feasible alternative for quantifying acute kinetic responses. Therefore, the purpose of this study was to evaluate the viability of a primed continuous oral sip-ingestion method of stable isotope tracers for the evaluation of whole-body protein kinetics. In a randomized, crossover design, eight healthy adults (63% female; Age: 29.4±5.8yrs; BMI: 24.3±2.7kg/m2) completed two, two-period stable isotope oral ingestion studies, consisting of a 3hbasal fasted period, followed by a 4-hpost-ingestion period. After the basal period, subjects ingested either 6.3g (Low) or 12.6g (High) of an essential amino acid (EAA) enriched whey protein supplement. The continuous oral sip-feed method was initiated with a primed oral bolus dose of L-[ring-2H5]phenylalanine, L-[ring-2H2]tyrosine, and L-[ring-2H4]tyrosine, followed by oral sip doses of L-[ring-2H5]phenylalanine, L-[ring-2H2]tyrosine every 10min to approximate steady state tracer enrichment. Blood samples were taken throughout the basal and post-meal periods to determine tracer enrichment. Whole-body net protein balance (NB), synthesis (PS), breakdown (PB), and exogenous hydroxylation were calculated for each period. Repeated measure ANOVAs (treatment×time) were used to assess differences in protein kinetics. Using the sip feed method, NB, PS, and hydroxylation were significantly increased with ingestion of protein (p<0.05) during the postprandial period, regardless of amount of protein ingested; ΔNB from the postabsorptive to postprandial period was significantly greater for high compared to low protein (p=0.026; low=6.2±5.1g protein·240min-1; high=11.8±3.9g protein·240min-1). The current study provides preliminary evidence that continuous oral sip-feeding of stable isotope tracer is a feasible method that provides physiologically relevant measures of protein metabolism. Assessments of variance and individual responses revealed high measurement variability with the sip-feed method compared to previously published constant infusion responses, but ΔNB, ΔPS, and ΔPB were comparable. In situations where constant infusion is not feasible, oral sip-feeding could be used as an alternative method for measurement of acute, postprandial protein metabolism.

Carbon sources and pathways for citrate secreted by human prostate cancer cells determined by NMR tracing and metabolic modeling

Prostate epithelial cells have the unique capacity to secrete large amounts of citrate, but the carbon sources and metabolic pathways that maintain this production are not well known. We mapped potential pathways for citrate carbons in the human prostate cancer metastasis cell lines LNCaP and VCaP, for which we first established that they secrete citrate (For LNCaP 5.6 ± 0.9 nmol/h per 106 cells). Using 13C-labeled substrates, we traced the incorporation of 13C into citrate by NMR of extracellular fluid. Our results provide direct evidence that glucose is a main carbon source for secreted citrate. We also demonstrate that carbons from supplied glutamine flow via oxidative Krebs cycle and reductive carboxylation routes to positions in secreted citrate but likely do not contribute to its net synthesis. The potential anaplerotic carbon sources aspartate and asparagine did not contribute to citrate carbons. We developed a quantitative metabolic model employing the 13C distribution in extracellular citrate after 13C glucose and pyruvate application to assess intracellular pathways of carbons for secreted citrate. From this model, it was estimated that in LNCaP about 21% of pyruvate entering the Krebs cycle is converted via pyruvate carboxylase as an anaplerotic route at a rate more than sufficient to compensate carbon loss of this cycle by citrate secretion. This model provides an estimation of the fraction of molecules, including citrate, leaving the Krebs cycle at every turn. The measured ratios of 13C atoms at different positions in extracellular citrate may serve as biomarkers for (malignant) epithelial cell metabolism.

Articulations and Products of Transition Systems and their Applications to Petri Net Synthesis

In order to speed up the synthesis of Petri nets from labelled transition systems, a divide and conquer strategy consists in defining decompositions of labelled transition systems, such that each component is synthesisable iff so is the original system. Then corresponding Petri Net composition operators are searched to combine the solutions of the various components into a solution of the original system. The paper presents two such techniques, which may be combined: products and articulations. They may also be used to structure transition systems, and to analyse the performance of synthesis techniques when applied to such structures.

Metabolic Homeostasis in Life as We Know It: Its Origin and Thermodynamic Basis.

Living organisms require continuous input of energy for their existence. As a result, life as we know it is based on metabolic processes that extract energy from the environment and make it available to support life (energy metabolism). This metabolism is based on, and regulated by, the underlying thermodynamics. This is important because thermodynamic parameters are stable whereas kinetic parameters are highly variable. Thermodynamic control of metabolism is exerted through near equilibrium reactions that determine. (1) the concentrations of metabolic substrates for enzymes that catalyze irreversible steps and (2) the concentrations of small molecules (AMP, ADP, etc.) that regulate the activity of irreversible reactions in metabolic pathways. The result is a robust homeostatic set point (−ΔGATP) with long term (virtually unlimited) stability. The rest of metabolism and its regulation is constrained to maintain this set point. Thermodynamic control is illustrated using the ATP producing part of glycolysis, glyceraldehyde-3-phosphate oxidation to pyruvate. Flux through the irreversible reaction, pyruvate kinase (PK), is primarily determined by the rate of ATP consumption. Change in the rate of ATP consumption causes mismatch between use and production of ATP. The resulting change in [ATP]/[ADP][Pi], through near equilibrium of the reactions preceding PK, alters the concentrations of ADP and phosphoenolpyruvate (PEP), the substrates for PK. The changes in ADP and PEP alter flux through PK appropriately for restoring equality of ATP production and consumption. These reactions appeared in the very earliest lifeforms and are hypothesized to have established the set point for energy metabolism. As evolution included more metabolic functions, additional layers of control were needed to integrate new functions into existing metabolism without changing the homeostatic set point. Addition of gluconeogenesis, for example, resulted in added regulation to PK activity to prevent futile cycling; PK needs to be turned off during gluconeogenesis because flux through the enzyme would waste energy (ATP), subtracting from net glucose synthesis and decreasing overall efficiency.

Characteristics of denitrification in a vertical baffled bioreactor

A vertical baffled bioreactor (VBBR) was employed for tertiary denitrification. Its features were designed to minimize the demand for externally supplied electron donor by minimizing net biomass synthesis and oxygen respiration. Over a two-year period, complete denitrification was realized routinely in the VBBR. The nitrate-removal rate was proportion to the influent COD/N ratio, with complete denitrification possible for COD/N ratios >3 gCOD/gN. Batch kinetic tests carried out at the end of years 1 and 2 documented that supplied electron donor was oxidized in the first 1–2 h, but nitrate and nitrite reductions occurred predominantly after 2 h and were driven by internally stored electron donor. Measurements confirmed that the VBBR minimized the demand of added electron donor: The observed yield was only 0.05 mgVSS/mgCOD, and the COD demand for O2 respiration was only 1–6.7% of the COD demand for N reductions. Biofilm samples taken from the upper and lower ports in cylinder of VBBR had similarly high alpha diversity and dominant genera, but the upper biofilm had a denitrification rate about 70% greater than the lower biofilm. The higher denitrification rate in the upper biofilm correlated its higher content of active biomass.

Evaluation of high protein intake to improve clinical outcome and nutritional status for patients with burns: a systematic review

Background and aimPatients with severe burns undergo a local and systemic response to the injury. As part of this response the patient becomes hypermetabolic. Current guidelines advise high protein intakes to counteract the catabolic response to burns, but this appears to be based on minimal experimental evidence. Hence the aim of this review was to examine the evidence for improvements in nutritional status and clinical outcome with the administration of high protein intakes for patients with burns. MethodsEight databases were searched for clinical trials with burn patients receiving two or more levels of protein intake at or above the level recommended for healthy individuals (0.75g/kg/d) and presenting results for at least one of the following pre-defined outcomes: nitrogen balance, length of stay, weight change, survival, physical therapy index, protein fractional synthetic rate, immunological measurements, bacteraemic days, systemic antibiotic days and net protein synthesis. ResultsSix studies were included, 4 of which were randomized trials. All had major methodological limitations, in particular none was blinded. There was too much heterogeneity in study design, patient characteristics and the timing and magnitude of the interventions to justify formal meta-analysis. There was no reliable evidence of improvement in nitrogen balance, but there was some evidence of increased weight gain on higher protein diets. One small study reported an increase in survival and significant improvements in infection rates and some indicators of immune function in children. Length of stay was not significantly improved. There was weak evidence of an improvement in muscle strength and endurance but no significant increase in protein synthesis in muscle or skin, or net protein synthesis in the whole body. ConclusionThere is currently only very weak evidence to justify administering high protein diets to patients following burns.

Protein Intake and Physical Performance Following Long-Term Stay on the International Space Station.

INTRODUCTION: Exposure to microgravity reduces muscle mass, volume, and performance. The ingestion of protein, especially combined with carbohydrate intake and exercise after ingestion, improves net muscle protein synthesis and increases muscle mass. However, there are few studies on this relationship during and after a long-term spaceflight. The objective of this study was to investigate the influence of protein and the combined effects of carbohydrate intake on muscle performance following long-term spaceflight.METHODS: This study is a retrospective cohort study involving secondary analysis of data stored in the NASA Lifetime Surveillance of Astronaut Health Repository. Multivariable analysis was performed to evaluate the impact of protein intake on physical performance by considering covariates potentially associated with each model.RESULTS: After adjusting for sex, age, flight week, energy intake, and preflight physical performance, protein intake was found to be significantly associated with concentric measurements for knee extension ( 51.66), ankle plantar flexion ( 32.86), and eccentric measurements for ankle plantar flexion ( 79.85) at 5 d after landing. Significant associations remained after controlling for exercise effect. No significant interaction between protein and carbohydrate intake was observed in either model.DISCUSSION: Protein intake during spaceflight was related to physical performance for knee extension and ankle plantar flexion, even after taking exercise effect into consideration. However, protein and carbohydrate intake provided no synergetic benefit. This suggests that high protein intake, about twice the current average intake, may serve as a countermeasure to offset the negative effects of long-duration spaceflights.Nozawa Y, Wagatsuma Y. Protein intake and physical performance following long-term stay on the International Space Station. Aerosp Med Hum Perform. 2021; 92(3):153159.

Rapid, -insensitive, dual-band quasi-adiabatic saturation transfer with optimal control for complete quantification of myocardial ATP flux.

PurposePhosphorus saturation‐transfer experiments can quantify metabolic fluxes noninvasively. Typically, the forward flux through the creatine kinase reaction is investigated by observing the decrease in phosphocreatine (PCr) after saturation of γ‐ATP. The quantification of total ATP utilization is currently underexplored, as it requires simultaneous saturation of inorganic phosphate (Pi) and PCr. This is challenging, as currently available saturation pulses reduce the already‐low γ‐ATP signal present.MethodsUsing a hybrid optimal‐control and Shinnar‐Le Roux method, a quasi‐adiabatic RF pulse was designed for the dual saturation of PCr and Pi to enable determination of total ATP utilization. The pulses were evaluated in Bloch equation simulations, compared with a conventional hard‐cosine DANTE saturation sequence, before being applied to perfused rat hearts at 11.7 T.ResultsThe quasi‐adiabatic pulse was insensitive to a >2.5‐fold variation in B1, producing equivalent saturation with a 53% reduction in delivered pulse power and a 33‐fold reduction in spillover at the minimum effective B1. This enabled the complete quantification of the synthesis and degradation fluxes for ATP in 30‐45 minutes in the perfused rat heart. While the net synthesis flux (4.24 ± 0.8 mM/s, SEM) was not significantly different from degradation flux (6.88 ± 2 mM/s, P = .06) and both measures are consistent with prior work, nonlinear error analysis highlights uncertainties in the Pi‐to‐ATP measurement that may explain a trend suggesting a possible imbalance.ConclusionsThis work demonstrates a novel quasi‐adiabatic dual‐saturation RF pulse with significantly improved performance that can be used to measure ATP turnover in the heart in vivo.