Accumulation Of By-products Research Articles

High-yield production of β-arbutin by identifying and eliminating byproducts formation.

β-Arbutin is a plant-derived glycoside and widely used in cosmetic and pharmaceutical industries because of its safe and effective skin-lightening property as well as anti-oxidant, anti-microbial, and anti-inflammatory activities. In recent years, microbial fermentation has become a highly promising method for the production of β-arbutin. However, this method suffers from low titer and low yield, which has become the bottleneck for its widely industrial application. In this study, we used β-arbutin to demonstrate methods for improving yields for industrial-scale production in Escherichia coli. First, the supply of precursors phosphoenolpyruvate and uridine diphosphate glucose was improved, leading to a 4.6-fold increase in β-arbutin production in shaking flasks. The engineered strain produced 36.12g/L β-arbutin with a yield of 0.11g/g glucose in a 3-L bioreactor. Next, based on the substrate and product's structural similarity, an endogenous O-acetyltransferase was identified as responsible for 6-O-acetylarbutin formation for the first time. Eliminating the formation of byproducts, including 6-O-acetylarbutin, tyrosine, and acetate, resulted in an engineered strain producing 43.79g/L β-arbutin with a yield of 0.22g/g glucose in fed-batch fermentation. Thus, the yield increased twofold by eliminating byproducts formation. To the best of our knowledge, this is the highest titer and yield of β-arbutin ever reported, paving the way for the industrial production of β-arbutin. This study demonstrated a systematic strategy to alleviate undesirable byproduct accumulation and improve the titer and yield of target products. KEY POINTS: •A systematic strategy to improve titer and yield was showed •Genes responsible for 6-O-acetylarbutin formation were firstly identified •43.79g/L β-arbutin was produced in bioreactor, which is the highest titer so far.

Removal of endocrine disruptor compounds, CO2 fixation, and macromolecules accumulation in Thermosynechococcus sp. CL-1 cultivation.

Recently, concern on several environmental issues including the pollutant discharge and high concentration of CO2 have gained high interest due to its impact on ecosystem and global warming effect, respectively. Implementation of photosynthetic microorganism carries out numerous advantages including high efficiency of CO2 fixation, the great endurance under extreme conditions and generation of valuable bioproducts. Thermosynechococcus sp. CL-1 (TCL-1), a cyanobacterium, has the ability to perform CO2 fixation and accumulation of various byproducts under extreme conditions like high temperature and alkalinity, presence of estrogen, or even using swine wastewater. This study aimed to assess TCL-1 performance under various endocrine disruptor compounds (bisphenol-A, 17-β-estradiol/E2, and 17-α-ethynilestradiol/EE2), concentrations (0-10mg/L), light intensities (500-2000 µE/m2/s), and dissolved inorganic carbon/DIC levels (0-113.2mM). Addition of E2 content even until 10mg/L carried out insignificant biomass growth interruption along with the improvement in CO2 fixation rate (79.8±0.1mg/L/h). Besides the influence of E2, application of higher DIC level and light intensity also enhanced the CO2 fixation rate and biomass growth. The highest biodegradation of E2 at 71% was achieved by TCL-1 in the end of 12h cultivation period. TCL-1 dominantly produced protein (46.7%±0.2%), however, production of lipid and carbohydrate (39.5±1.5 and 23.3±0.9%, respectively) also could be considered as the potential source for biofuel production. Thus, this study can provide an efficient strategy in simultaneously dealing with environmental issues with side advantage in production of macromolecules.

Increasing power output and isopropanol biodegradation by trickle-bed microbial fuel cells by optimizing air relative humidity and oxygen flux at cathode

The cathode air flow rate and relative humidity (RH) of a trickle-bed microbial fuel cell (TB-MFC) were optimized using the response surface methodology to obtain a high oxygen flux and a low electrical resistance at the cathode and thereby promote the oxygen reduction reaction. Under optimal air flow rate (2.20 L/min) and RH (20%) conditions, the TB-MFC system exhibited the highest oxygen flux (3.08%/cm2‧min) and the lowest resistance (5.33 Ω); its regression model had an R2 > 0.95 and the desirability coefficient was 0.78. These results demonstrate that the optimized air flow rate and RH support high oxygen flux and low resistance at the cathode. Using biochar combined with conductive carbon black (CCB) as the cathodic catalyst and optimal cathodic operating conditions, yielded a mineralization efficiency of isopropanol of 96.9% and a power density of 555 mW/m3 at an empty bed residence time and isopropanol influx concentration of 60 s and 300 ppmv, respectively. These results suggest that isopropanol can be effectively mineralized directly into CO2 and H2O by a TB-MFC, reducing the by-product accumulation, and thus increasing the power generation of the TB-MFC. Optimizing the air flow rate and RH of the biochar/CCB-based cathode in the TB-MFC increases the oxygen flux and reduces the electrical resistance of the cathode of TB-MFC, which can thus be used to treat exhaust gases that contain organics.

Study on the construction technology of β-alanine synthesizing Escherichia coli based on cellulosome assembly.

Introduction: β-Alanine is the only β-amino acid in nature; it is widely used in food additives, medicines, health products, and surfactants. To avoid pollution caused by traditional production methods, the synthesis of β-alanine has been gradually replaced by microbial fermentation and enzyme catalysis, which is a green, mild, and high-yield biosynthesis method. Methods: In this study, we constructed an Escherichia coli recombinant strain for efficient β-alanine production using glucose as the raw material. The microbial synthesis pathway of L-lysine-producing strain, Escherichia coli CGMCC 1.366, was modified using gene editing by knocking out the aspartate kinase gene, lysC. The catalytic efficiency and product synthesis efficiency were improved by assembling key enzymes with cellulosome. Results: By-product accumulation was reduced by blocking the L-lysine production pathway, thereby increasing the yield of β-alanine. In addition, catalytic efficiency was improved by the two-enzyme method to further increase the β-alanine content. The key cellulosome elements, dockerin (docA) and cohesin (cohA), were combined with L-aspartate-α-decarboxylase (bspanD) from Bacillus subtilis and aspartate aminotransferase (aspC) from E.coli to improve the catalytic efficiency and expression level of the enzyme. β-alanine production reached 7.439mg/L and 25.87mg/L in the two engineered strains. The β-alanine content reached 755.465mg/L in a 5L fermenter. Discussion: The content of β-alanine synthesized by constructed β-alanine engineering strains were 10.47 times and 36.42 times higher than the engineered strain without assembled cellulosomes, respectively. This research lays the foundation for the enzymatic production of β-alanine using a cellulosome multi-enzyme self-assembly system.

Nano iron tetroxide-modified rice husk biochar promoted Feammox performance of Klebsiella sp. FC61 and synergistically removed Ni2+ and ciprofloxacin

The iron reduction coupled with ammonia oxidation process (Feammox) is a biological reaction process associated with the nitrogen cycle that has been discovered in recent years. In this study, the iron-reducing bacterium Klebsiella sp. FC61 was attached by synthesizing nano-loadings of iron tetroxide (nFe3O4) onto rice husk biochar (RBC), and the RBC-nFe3O4 was used as an electron shuttle to participate in the biological iron reduction process of soluble and insoluble Fe3+ to improve the ammonia oxidation efficiency to 81.82%. This acceleration of electron transfer increased the carbon consumption rate and further tuned up the COD removal efficiency to 98.00%. The Feammox could be coupled with iron denitrification for internal nitrogen/iron cycling to reduce the accumulation of nitrate by-products and achieve the recycling of iron. In addition, pollutants such as Ni2+, ciprofloxacin, and formed chelates could be removed by pore adsorption and π-π interactions using bio-iron precipitates produced by iron-reducing bacteria.

Irreversible biosynthesis of D-Allulose from D-glucose in Escherichia coli through fine-tuning of carbon flux and cofactor regeneration engineering.

As a rare hexose with low calories and various physiological functions, D-allulose has drawn increasing attention. The current industrial production of D-allulose from D-fructose or D-glucose is achieved via epimerization based on the Izumoring strategy; however, the inherent reaction equilibrium during reversible reaction limits its high conversion yield. Although the conversion of D-fructose to D-allulose could be enhanced via phosphorylation-dephosphorylation mediated by metabolic engineering, biomass reduction and byproduct accumulation remain a largely unresolved issue. After modifying the glycolytic pathway of Escherichia coli and optimizing the whole-cell reaction condition, the engineered strain produced 7.57 ± 0.61 g L-1 D-allulose from 30 g L-1 D-glucose after 24 h of catalysis. By developing an ATP regeneration system for enhanced substrate phosphorylation, the cell growth inhibition was alleviated and D-allulose production increased by 55.3% to 11.76 ± 0.58 g L-1 (0.53 g g-1 ). Fine-tuning of carbon flux caused a 48% reduction in D-fructose accumulation to 1.47 ± 0.15 g L-1 . After implementing fed-batch co-substrate strategy, the D-allulose titer reached 15.80 ± 0.31 g L-1 (0.62 g g-1 ) with a D-glucose conversion rate of 84.8%. This study presented a novel strategy for high-yield D-allulose production from low-cost substrate. This article is protected by copyright. All rights reserved.

New insights into the mechanism of Fered-Fenton treatment of industrial wastewater with high chloride content: Role of multiple reactive species

Hydroxyl radical (OH) is considered the dominant reactive species in the electro-Fenton (EF) and Fered-Fenton (EF-Fere) processes for wastewater treatment. However, in chloride-rich media, this is arguable due to the obscure mechanisms for the oxidant speciation and pollutant degradation. Herein, the role of active chlorine and Fe(IV)-oxo species (FeIVO2+) as primary oxidizing agents in HClO-mediated Fered-Fenton (EF-Fere-HClO) process is discussed, along with the dependence of their contribution on the pollutant structure. HClO generated from anodic oxidation of Cl− can be consumed by added H2O2 to form singlet oxygen (1O2), which is detrimental because this species is quickly deactivated by water. The reaction between HClO and Fe2+ was proved to generate FeIVO2+, rather than OH or Cl suggested in the literature. The yield of FeIVO2+ species was proportional to the Cl− concentration and barely affected by solution pH. The long-lived HClO and FeIVO2+ can selectively react with electron-rich compounds, which occurs simultaneously to the non-selective attack of OH formed from Fenton's reaction. The FeIVO2+ and OH concentration profiles were successfully modelled. Although the accumulation of toxic chlorinated by-products from HClO-mediated oxidation might cause new environmental concerns, the toxicity of pesticide wastewater with 508 mM Cl− was halved upon EF-Fere-HClO treatment.

Concurrent metaboreflex activation increases chronotropic and ventilatory responses to passive leg movement without sex-related differences

Previous studies in animal models showed that exercise-induced metabolites accumulation may sensitize the mechanoreflex-induced response. The aim of this study was to assess whether the magnitude of the central hemodynamic and ventilatory adjustments evoked by isolated stimulation of the mechanoreceptors in humans are influenced by the prior accumulation of metabolic byproducts in the muscle. 10 males and 10 females performed two exercise bouts consisting of 5-min of intermittent isometric knee-extensions performed 10% above the previously determined critical force. Post-exercise, the subjects recovered for 5 min either with a suprasystolic circulatory occlusion applied to the exercised quadriceps (PECO) or under freely-perfused conditions (CON). Afterwards, 1-min of continuous passive leg movement was performed. Central hemodynamics, pulmonary data, and electromyography from exercising/passively-moved leg were recorded throughout the trial. Root mean square of successive differences (RMSSD, index of vagal tone) was also calculated. Δpeak responses of heart rate (ΔHR) and ventilation (Delta dot{V}_{{text{E}}}) to passive leg movement were higher in PECO compared to CON (ΔHR: 6 ± 5 vs 2 ± 4 bpm, p = 0.01; 3.9 ± 3.4 vs 1.9 ± 1.7 L min−1, p = 0.02). Δpeak of mean arterial pressure (ΔMAP) was significantly different between conditions (5 ± 3 vs − 3 ± 3 mmHg, p < 0.01). Changes in RMSSD with passive leg movement were different between PECO and CON (p < 0.01), with a decrease only in the former (39 ± 18 to 32 ± 15 ms, p = 0.04). No difference was found in all the other measured variables between conditions (p > 0.05). These findings suggest that mechanoreflex-mediated increases in HR and dot{V}_{{text{E}}} are sensitized by metabolites accumulation. These responses were not influenced by biological sex.

The Involvement of Glucose and Lipid Metabolism Alteration in Rheumatoid Arthritis and Its Clinical Implication.

Obviously, immune cells like T cells and macrophages play a major role in rheumatoid arthritis (RA). On one hand, the breakdown of immune homeostasis directly induces systemic inflammation; on the other hand, these cells initiate and perpetuate synovitis and tissue damages through the interaction with fibroblast-like synoviocytes (FLS). In recent years, the pathological link between metabolic disorders and immune imbalance has received increasing attention. High energy demand of immune cells leads to the accumulation of metabolic byproducts and inflammatory mediators. They act on various metabolism-sensitive signal pathways as well as relevant transcription factors, such as HIF-1α, and STATs. These molecular events will impact RA-related effectors like circulating immune cells and joint-resident cells in return, allowing the continuous progression of systemic inflammation, arthritic manifestations, and life-threatening complications. In other words, metabolic complications are secondary pathological factors for the progression of RA. Therefore, the status of energy metabolism may be an important indicator to evaluate RA severity, and in-depth explorations of the mechanisms underlying the mystery of how RA-related metabolic disorders develop will provide useful clues to further clarify the etiology of RA, and inspire the discovery of new anti-rheumatic targets. This article reviews the latest research progress on the interactions between immune and metabolism systems in the context of RA. Great importance is attached to the changes in certain pathways controlling both immune and metabolism functions during RA progression.

N20D/N116E Combined Mutant Downward Shifted the pH Optimum of Bacillus subtilis NADH Oxidase.

Cofactor regeneration is indispensable to avoid the addition of large quantities of cofactor NADH or NAD+ in oxidation-reduction reactions. Water-forming NADH oxidase (Nox) has attracted substantive attention as it can oxidize cytosolic NADH to NAD+ without concomitant accumulation of by-products. However, its applications have some limitations in some oxidation-reduction processes when its optimum pH is different from its coupled enzymes. In this study, to modify the optimum pH of BsNox, fifteen relevant candidates of site-directed mutations were selected based on surface charge rational design. As predicted, the substitution of this asparagine residue with an aspartic acid residue (N22D) or with a glutamic acid residue (N116E) shifts its pH optimum from 9.0 to 7.0. Subsequently, N20D/N116E combined mutant could not only downshift the pH optimum of BsNox but also significantly increase its specific activity, which was about 2.9-fold at pH 7.0, 2.2-fold at pH 8.0 and 1.2-fold at pH 9.0 that of the wild-type. The double mutant N20D/N116E displays a higher activity within a wide range of pH from 6 to 9, which is wider than the wide type. The usability of the BsNox and its variations for NAD+ regeneration in a neutral environment was demonstrated by coupling with a glutamate dehydrogenase for α-ketoglutaric acid (α-KG) production from L-glutamic acid (L-Glu) at pH 7.0. Employing the variation N20D/N116E as an NAD+ regeneration coenzyme could shorten the process duration; 90% of L-Glu were transformed into α-KG within 40 min vs. 70 min with the wild-type BsNox for NAD+ regeneration. The results obtained in this work suggest the promising properties of the BsNox variation N20D/N116E are competent in NAD+ regeneration applications under a neutral environment.

Enhanced 1,3-propanediol production with high yield from glycerol through a novel Klebsiella–Shewanella co-culture

Background1,3-Propanediol (1,3-PDO) is a platform compound, which has been widely used in food, pharmaceutical and cosmetic industries. Compared with chemical methods, the biological synthesis of 1,3-PDO has shown promising applications owing to its mild conditions and environmental friendliness. However, the biological synthesis of 1,3-PDO still has the problem of low titer and yield due to the shortage of reducing powers.ResultsIn this study, Klebsiella sp. strain YT7 was successfully isolated, which can synthesize 11.30 g/L of 1,3-PDO from glycerol in flasks. The intracellular redox regulation strategy based on the addition of electron mediators can increase the 1,3-PDO titer to 28.01 g/L. Furthermore, a co-culturing system consisting of strain YT7 and Shewanella oneidensis MR-1 was established, which can eliminate the supplementation of exogenous electron mediators and reduce the by-products accumulation. The 1,3-PDO yield reached 0.44 g/g and the final titer reached 62.90 g/L. The increased titer and yield were attributed to the increased redox levels and the consumption of by-products.ConclusionsA two-bacterium co-culture system with Klebsiella sp. strain YT7 and S. oneidensis strain MR-1 was established, which realized the substitution of exogenous electron mediators and the reduction of by-product accumulation. Results provided theoretical basis for the high titer of 1,3-PDO production with low by-product concentration.

The Glymphatic System May Play a Vital Role in the Pathogenesis of Hepatic Encephalopathy: A Narrative Review

Hepatic encephalopathy (HE) is a neurological complication of liver disease resulting in cognitive, psychiatric, and motor symptoms. Although hyperammonemia is a key factor in the pathogenesis of HE, several other factors have recently been discovered. Among these, the impairment of a highly organized perivascular network known as the glymphatic pathway seems to be involved in the progression of some neurological complications due to the accumulation of misfolded proteins and waste substances in the brain interstitial fluids (ISF). The glymphatic system plays an important role in the clearance of brain metabolic derivatives and prevents aggregation of neurotoxic agents in the brain ISF. Impairment of it will result in aggravated accumulation of neurotoxic agents in the brain ISF. This could also be the case in patients with liver failure complicated by HE. Indeed, accumulation of some metabolic by-products and agents such as ammonia, glutamine, glutamate, and aromatic amino acids has been reported in the human brain ISF using microdialysis technique is attributed to worsening of HE and correlates with brain edema. Furthermore, it has been reported that the glymphatic system is impaired in the olfactory bulb, prefrontal cortex, and hippocampus in an experimental model of HE. In this review, we discuss different factors that may affect the function of the glymphatic pathways and how these changes may be involved in HE.

High-level itaconic acid (IA) production using engineered Escherichia coli Lemo21(DE3) toward sustainable biorefinery

Itaconic acid (IA) serves as a prominent building block for polyamides as sustainable material. In vivo IA production is facing the competing side reactions, byproducts accumulation, and long cultivation time. Therefore, the utilization of whole-cell biocatalysts to carry out production from citrate is an alternative approach to sidestep the current limitations. In this study, in vitro reaction of IA was obtained 72.44 g/L by using engineered E. coli Lemo21(DE3) harboring the aconitase (Acn, EC 4.2.1.3) and cis-aconitate decarboxylase (CadA, EC 4.1.1.6) which was cultured in glycerol-based minimal medium. IA productivity enhancement was observed after cold-treating the biocatalysts in − 80 °C for 24 h prior to the reaction, reaching 81.6 g/L. On the other hand, a new seeding strategy in Terrific Broth (TB) as a nutritionally rich medium was employed to maintain the biocatalysts stability up to 30 days. Finally, the highest IA titer of 98.17 g/L was attained using L21::7G chassis, that has a pLemo plasmid and integration of GroELS to the chromosome. The high-level of IA production along with the biocatalyst reutilization enables the economic viability toward a sustainable biorefinery.

Improving the production of the micafungin precursor FR901379 in an industrial production strain

BackgroundMicafungin is an echinocandin-type antifungal agent used for the clinical treatment of invasive fungal infections. It is semisynthesized from the sulfonated lipohexapeptide FR901379, a nonribosomal peptide produced by the filamentous fungus Coleophoma empetri. However, the low fermentation efficiency of FR901379 increases the cost of micafungin production and hinders its widespread clinical application.ResultsHere, a highly efficient FR901379-producing strain was constructed via systems metabolic engineering in C. empetri MEFC09. First, the biosynthesis pathway of FR901379 was optimized by overexpressing the rate-limiting enzymes cytochrome P450 McfF and McfH, which successfully eliminated the accumulation of unwanted byproducts and increased the production of FR901379. Then, the functions of putative self-resistance genes encoding β-1,3-glucan synthase were evaluated in vivo. The deletion of CEfks1 affected growth and resulted in more spherical cells. Additionally, the transcriptional activator McfJ for the regulation of FR901379 biosynthesis was identified and applied in metabolic engineering. Overexpressing mcfJ markedly increased the production of FR901379 from 0.3 g/L to 1.3 g/L. Finally, the engineered strain coexpressing mcfJ, mcfF, and mcfH was constructed for additive effects, and the FR901379 titer reached 4.0 g/L under fed-batch conditions in a 5 L bioreactor.ConclusionsThis study represents a significant improvement for the production of FR901379 and provides guidance for the establishment of efficient fungal cell factories for other echinocandins.

NAD+ supplementation improves mAb productivity in CHO cells via a glucose metabolic shift.

Aerobic glycolysis and its by-product lactate accumulation are usually associated with adverse culture phenotypes such as poor cell viability and productivity. Due to the lack of knowledge on underlying mechanisms and accompanying biological processes, the regulation of aerobic glycolysis has been an ongoing challenge in culture process development for therapeutic protein productivity. Nicotinamide adenine dinucleotide (NAD+ ), a coenzyme and co-substrate in energy metabolism, promotes the conversion of inefficient glycolysis into an efficient oxidative phosphorylation (OXPHOS) pathway. However, the effect of NAD+ on Chinese hamster ovary (CHO) cells for biopharmaceutical production has not been reported yet. In this work, we aimed to elucidate the influence of NAD+ on cell culture performance by examining metabolic shifts and mAb productivity. The supplementation of NAD+ increased the intracellular concentration of NAD+ and promoted SIRT3 expression. Antibody titer and the specific productivity in the growth phase were improved by up to 1.82- and 1.88-fold, respectively, with marginal restrictions on cell growth. NAD+ significantly reduced the accumulation of reactive oxygen species (ROS) and the lactate yield from glucose, determined by lactate accumulation versus glucose consumption (YLAC/GLC ). In contrast, OXPHOS capacity and amino acid consumption rate increased substantially. Collectively, these results suggest that NAD+ contributes to improving therapeutic protein productivity in bioprocessing via inducing an energy metabolic shift.

Dynamically Interfacial pH-Buffering Effect Enabled by N-Methylimidazole Molecules as Spontaneous Proton Pumps toward Highly Reversible Zinc-Metal Anodes.

Aqueous zinc-metal batteries have attracted extensive attention due to their outstanding merits of high safety and low cost. However, the intrinsic thermodynamic instability of zinc in aqueous electrolyte inevitably results in hydrogen evolution, and the consequent generation of OH- at the interface will dramatically exacerbate the formation of dead zinc and dendrites. Herein, a dynamically interfacial pH-buffering strategy implemented by N-methylimidazole (NMI) additive is proposed to remove the detrimental OH- at zinc/electrolyte interface in real-time, thus eliminating the accumulation of by-products fundamentally. Electrochemical quartz crystal microbalance and molecular dynamics simulation results reveal the existence of an interfacial absorption layer assembled by NMI and protonated NMI (NMIH+ ), which acts as an ion pump for replenishing the interface with protons constantly. Moreover, an in situ interfacial pH detection method with micro-sized spatial resolution based on the ultra-microelectrode technology is developed to probe the pH evolution in diffusion layer, confirming the stabilized interfacial chemical environment in NMI-containing electrolyte. Accordingly, with the existence of NMI, an excellent cumulative plating capacity of 4.2 Ah cm-2 and ultrahigh Coulombic efficiency of 99.74% are realized for zinc electrodes. Meanwhile, the NMI/NMIH+ buffer additive can accelerate the dissolution/deposition process of MnO2 /Mn2+ on the cathode, leading to enhanced cycling capacity.

Silver decorated silanized graphene oxide based molecularly surface imprinted electrochemical sensor for the trace level detection of L- Tryptophan

L-Tryptophan (TRP) is a major precursor for neurotransmitter Serotonin, neurohormone Melatonin, Vitamin B3, phytohormone Auxins, Catecholamine Adrenalin, Dopamine, and Melamine. It has some adverse effects like the production of toxic substances in the brain leading to Hallucination Delusion, Schizophrenia, Parkinson's disease, and Alzheimer's disease. The oxidative by-product accumulation also leads to the generation of cancer. Due to its importance as well as toxic effects, there is an increased significance for the accurate determination of TRP. Silver decorated Silanized graphene oxide (Silanized Ag-GO) based molecularly imprinted electrochemical sensor was synthesized and used for the selective and specific detection of TRP. Silyl functionalization was done by using vinyl trimethoxysilane (VTMS). Pre-polymerization mixture was prepared by mixing the template TRP and the monomers such as methacrylic acid (MAA) and styrene in dimethyl sulfoxide (DMSO). The electrochemical properties of the synthesized materials were characterized using Electrochemical Impedance Spectroscopy (EIS), Cyclic Voltammetry (CV), and Differential Pulse Voltammetry (DPV). Working electrodes were prepared by modifying the Bare Glassy Carbon electrode (Bare GCE) with TRP imprinted polymer (TRP/MIP) and non-imprinted polymer (TRP/NIP). The surface area of TRP/NIP, TRP/MIP modified Bare GCE electrodes (TRP/NIP/GCE and TRP/MIP/GCE) was calculated as 0.2437, and 0.443 cm2 respectively. The limit of detection (LOD) and limit of quantification (LOQ) were calculated as 3.23 × 10−10 M and 10.76 × 10−10 M with a correlation coefficient (R2) value of 0.9958. In addition to this, herein reported electrochemical sensor was successfully used for the detection of TRP in the blood serum sample. This exposes the perspective clinical application of the synthesized electrochemical sensor.

Enhancing sustainability within industrial cooperative networks through the evaluation of economically compromised entities

IntroductionWhile promoting sustainable industrial cluster development using the circular economy perspective, the increased use of underutilized materials results in compromised profitability in the cooperative network. The focus is to evaluate the external financial support required in industrial clusters against specific objectives and highlight the potential beneficiaries and losers in financial terms because of industrial symbiosis while closing material loops.MethodThis study provides an agent-based approach to record the system response based on three case studies to demonstrate the potential cost variations: The first case is about utilizing only naturally available resources by all the industrial entities and is taken as a base case. The second case is about targeting the lowest cost for each product, and the third one provides a sustainable and flexible solution by targeting the best transformation methods and materials.ConclusionThe study concludes with valuable insights to identify the economically compromised entities in an industrial cluster network by considering economic deviations beyond a critical value. It can help take concrete measures in the form of incentives or investors subsidies by governmental organizations, regulators, and policymakers to intervene and stimulate markets through targeted financial support/policies to the compromised entities. This results in improved materials loop closing, essentially promoting sustainable production systems in industrial clusters. Additionally, such financial support/incentives also influence the scarcity or accumulation of by-products or low-value materials, ultimately improving the industrial network's environmental and economic performance.

Partners in crime: The feedback loop between metabolic reprogramming and immune checkpoints in the tumor microenvironment.

The tumor microenvironment (TME) is a complex and constantly changing cellular system composed of heterogeneous populations of tumor cells and non-transformed stromal cells, such as stem cells, fibroblasts, endothelial cells, pericytes, adipocytes, and innate and adaptive immune cells. Tumor, stromal, and immune cells consume available nutrients to sustain their proliferation and effector functions and, as a result of their metabolism, produce a wide array of by-products that gradually alter the composition of the milieu. The resulting depletion of essential nutrients and enrichment of by-products work together with other features of the hostile TME to inhibit the antitumor functions of immune cells and skew their phenotype to promote tumor progression. This review briefly describes the participation of the innate and adaptive immune cells in recognizing and eliminating tumor cells and how the gradual metabolic changes in the TME alter their antitumor functions. In addition, we discuss the overexpression of the immune checkpoints and their ligands as a result of nutrient deprivation and by-products accumulation, as well as the amplification of the metabolic alterations induced by the immune checkpoints, which creates an immunosuppressive feedback loop in the TME. Finally, the combination of metabolic and immune checkpoint inhibitors as a potential strategy to treat cancer and enhance the outcome of patients is highlighted.

SGLT2 inhibitors: role in protective reprogramming of cardiac nutrient transport and metabolism.

Sodium-glucose cotransporter 2 (SGLT2) inhibitors reduce heart failure events by direct action on the failing heart that is independent of changes in renal tubular function. In the failing heart, nutrient transport into cardiomyocytes is increased, but nutrient utilization is impaired, leading to deficient ATP production and the cytosolic accumulation of deleterious glucose and lipid by-products. These by-products trigger downregulation of cytoprotective nutrient-deprivation pathways, thereby promoting cellular stress and undermining cellular survival. SGLT2 inhibitors restore cellular homeostasis through three complementary mechanisms: they might bind directly to nutrient-deprivation and nutrient-surplus sensors to promote their cytoprotective actions; they can increase the synthesis of ATP by promoting mitochondrial health (mediated by increasing autophagic flux) and potentially by alleviating the cytosolic deficiency in ferrous iron; and they might directly inhibit glucose transporter type 1, thereby diminishing the cytosolic accumulation of toxic metabolic by-products and promoting the oxidation of long-chain fatty acids. The increase in autophagic flux mediated by SGLT2 inhibitors also promotes the clearance of harmful glucose and lipid by-products and the disposal of dysfunctional mitochondria, allowing for mitochondrial renewal through mitochondrial biogenesis. This Review describes the orchestrated interplay between nutrient transport and metabolism and nutrient-deprivation and nutrient-surplus signalling, to explain how SGLT2 inhibitors reverse the profound nutrient, metabolic and cellular abnormalities observed in heart failure, thereby restoring the myocardium to a healthy molecular and cellular phenotype.