Activity Of Coenzyme Research Articles

Pathological and Hormonal Alterations of the Testis in Balb/C Mice Treated With Spirotetramat Insecticide

Background and Objectives: The side effects of insecticides are undeniable due to their excessive use by farmers. This study aims to investigate the impact of spirotetramat insecticide on pathological and hormonal alteration in BALB/c mice testes. Methods: In this experimental study, 24 adult BALB/c male mice, with a body weight range of 25-30 g, were obtained from the Animal House of Uremia University and examined. Mice were randomly divided into three groups of eight: control, experimental group 1, and experimental group 2. The mice of experimental group 1 received 2.5 mg/kg, and experimental group 2 received 10 mg/kg of spirotetramat. The control group received the same amount of distilled water for 21 days through the mouth by gavage. After this period, the mice were anesthetized and evaluated. Data were analyzed using a 1-way analysis of variance in SPSS version 19 software. Results: The results of the study showed that spirotetramat increased the thickness of the tunica albuginea, the diameter of the seminiferous tubules, the differentiation index of the testis tube, and the spermiogenesis index, but it decreased the thickness of the epithelium of the spermatogenic tubules, the replacement index of active spermatogonia, Leydig cells, and FSH, LH, and testosterone hormone levels. The thickness of interstitial tissue, Sertoli, and lymphocyte cells did not change significantly. Conclusion: By inhibiting the activity of acetyl coenzyme A, spirotetramat insecticide disrupts energy production and prevents the synthesis of lipids in the cells, finally decreasing spermatogenesis in mice.

Seaweed as a feed additive to mitigate enteric methane emissions in ruminants: Opportunities and challenges1

Cutting farming-related methane emissions from ruminants is critical in the battle against climate change. Since scientists initially investigated the potential of marine macroalgae to reduce methane emissions, using seaweeds as an anti-methanogenic feed additive has become prevailing in recent years. Asparagopsis taxiformis is the preferred species because it contains a relatively higher concentration of bromoform. As a type of halogenated methane analogue, bromoform contained in A. taxiformis can specifically inhibit the activity of coenzyme M methyltransferase, thereby blocking the ruminal methanogenesis. However, bromoform is a potential toxin and ozone-depleting substance. In response, current research focuses on the effects of bromoform-enriched seaweed supplementation on ruminant productivity and safety, as well as the impact of large-scale cultivation of seaweeds on the atmospheric environment. The current research on seaweed still needs to be improved, especially in developing more species with low bromoform content, such as Bonnemaisonia hamifera, Dictyota bartayresii, and Cystoseira trinodis. Otherwise, seaweed is rich in bioactive substances and exhibits antibacterial, anti-inflammatory, and other physiological properties, but research on the role of these bioactive compounds in methane emissions is lacking. It is worthy of deeper investigation to identify more potential bioactive compounds. As a new focus of attention, seaweed has attracted the interest of many scientists. Nevertheless, seaweed still faces some challenges as a feed additive to ruminants, such as the residues of heavy metals (iodine and bromine) and bromoform in milk or meat, as well as the establishment of a supply chain for seaweed cultivation, preservation, and processing. We have concluded that the methane-reducing efficacy of seaweed is indisputable. However, its application as a commercial feed additive is still influenced by factors such as safety, costs, policy incentives, and regulations.

Nicotinamide Adenine Dinucleotide: The Redox Sensor in Aging-Related Disorders.

Significance: Nicotinamide adenine dinucleotide (NADH) represents the reduced form of NAD+, and together they constitute the two forms of the nicotinamide adenine dinucleotide whose balance is named as the NAD+/NADH ratio. NAD+/NADH ratio is mainly involved in redox reactions since both the molecules are responsible forcarrying electrons to maintain redox homeostasis. Recent Advances: NADH acts as a reducing agent, and one of the most known processes exploiting NADH function is energy metabolism. The two main pathways generating energy and involving NADH are glycolysis and oxidative phosphorylation, occurring in cell cytosol and in the mitochondrial matrix, respectively. Critical Issues: Although NADH is primarily produced through the reduction of NAD+ and consumed by its own oxidation, several are the biosynthetic and consumption pathways, reflecting the NADH role in multiple cellular processes. This review gathers all the main current data referring to NADH incorrelation with metabolic and cellular pathways, such as its coenzyme activity, effect in cell death, and on modulating redox and calcium homeostasis. Future Directions: Gene expression control, as well as the potential impact on neurodegenerative, cardiac disorders and infections, suggest NADH application in clinical settings.Thorough clinical trials and continued investigation into the long-term impacts of NADH are crucial to validate its effectiveness and safety, thereby facilitating its wider acceptance as a therapeutic option in medical practice.

Application of Aptamer-SERS Nanotags for Unveiling the PD-L1 Immunomarker Progression Correlated to the Cell Metabolic Bioprocess.

In recent years, the expression and progression of programmed cell death ligand 1 (PD-L1) as an immunomarker in the context of a cell metabolic environment has gained significant attention in cancer research. However, intercellular bioprocesses that control the dynamics of PD-L1 have been largely unexplored. This study aimed to explore the cell metabolic states and conditions that govern dynamic variations of PD-L1 within the cell metabolic environment using an aptamer-based surface-enhanced Raman scattering (SERS) approach. The aptamer-SERS technique offers a sensitive, rapid, and powerful analytical tool for targeted and nondestructive detection of an immunomarker with high sensitivity and specificity. By combining aptamer-SERS with cell state profiling, we investigated the modulation in PD-L1 expression under different metabolic states, including glucose deprivation, metabolic coenzyme activity, and altered time/concentration-based cytokine availability. The most intriguing features in our findings include the cell-specific responses, cell differentiation by revealing distinct patterns, and dynamics of PD-L1 in different cell lines. Additionally, the time-dependent variations in PD-L1 expression, coupled with the dose-dependent relationship between glucose concentration and PD-L1 levels, underscore the complex interplay between immune checkpoint regulation and cellular metabolism. Therefore, this work demonstrates the advantages of using highly-sensitive and specific aptamer-SERS nanotags for investigating the immune checkpoint dynamics and related metabolic bioprocess.

Mechanisms of anaerobic treatment of sulfate-containing organic wastewater mediated by Fe0 under different initial pH values.

The anaerobic treatment of sulfide-containing organic wastewater (SCOW) is significantly affected by pH, causing dramatic decrease of treatment efficiency when pH deviates from its appropriate range. Fe0 has proved as an effective strategy on mitigating the impact of pH. However, systematic analysis of the influence mechanism is still lacking. To fill this gap, the impact of different initial pH values on anaerobic treatment efficiency of SCOW with Fe0 addition, the change of fermentation type and methanogens, and intra-extracellular electron transfer were explored in this study. The results showed that Fe0 addition enhanced the efficacy of anaerobic treatment of SCOW at adjusted initial pH values, especially at pH 6. Mechanism analysis showed that respiratory chain-related enzymes and electron shuttle secretion and resistance reduction were stimulated by soluble iron ions generated by Fe0 at pH 6, which accelerated intra-extracellular electron transfer of microorganisms, and ultimately alleviated the impact of acidic pH on the system. While at pH 8, Fe0 addition increased the acetogenic bacteria abundance, as well as optimized the fermentation type and improved the F420 coenzyme activity, resulting in the enhancement of treatment efficiency in the anaerobic system and remission of the effect of alkaline pH on the system. At the neutral pH, Fe0 addition had both advantages as stimulating the secretion ofrespiratory chainand electron transfer-related enzymes at pH 6 and optimizing the fermentation type pH 8, and thus enhanced the treatment efficacy. This study provides important insights and scientific basis for the application of new SCOW treatment technologies.

Shading in fruit changes the polyphenol accumulation of pellicle by regulating activity of key enzymes and expression of their gene related polyphenol anabolism of Juglans sigillata Dode

Polyphenols, abundant in the pellicle of walnuts, confer health benefits to the human body. The intricate relationship between light and phenolic compounds underscores the impact of shading, which can effectively diminish phenolic content and enhance the palatability of food. In this study, we delved into the connection between polyphenol accumulation, related enzyme activities, and gene expression in the pellicle of Juglans sigillata Dode under varying degrees of shading. The results show that different shade intensities can change polyphenols’ content and affect the activity of key polyphenols anabolism enzymes and their gene expression. With the increase of the shading intensity, the total phenolics content (TPC) and total flavonoids content (TFC) were lower than the nomal light. In the mature stage, shading 60 % was lowest. 7 individual phenolics were identified in the pellicle, and the content of each varied significantly with the decrease of light intensity. Except for p-coumaric acid and ferulic acid, which were higher than the control at mature stage, all phenolics were lower than the control, but the overall trend was the same as that of the control. The activity of key enzymes in the anabolism of polyphenols was significantly affected after shading. In complete darkness phenylalanine ammonia lyase (PAL) was highest at the kernel-filling stage, shading 20 % of chalcone synthase (CHS) and shading 60 % of flavonol synthase (FLS) responded most strongly at the hard-core stage, while flavanone 3-hydroxylase (F3H) was most active at shading 60 % of kernel-filling stage, which was 2.03, 1.38, 4.67, and 2.08 times that of the control, respectively. The expression levels of key genes showed that the expression level of JsPAL was significantly increased at hard-core stage and mature stage, and the expression of JsPAL was 3.85 times higher than that of the control at the mature stage with 60 % shading. Correlation analysis showed that PAL enzyme activity was highly correlated with TPC, TFC and various individual phenolics with the increase of shading intensity. The content of p-coumaric acid was positively correlated with the activities of 4-coumarate coenzyme A ligase (4CL), F3H and FLS. The increase of shading intensity enhanced the correlation between total phenolics content, total flavonoids content, individual phenolics substances and JsPAL, JsC4H, JsCHS, JsCHI and JsFLS. Overall, PAL was a key enzyme affecting polyphenol accumulation under different shading levels, while at the genetic level, JsPAL, JsC4H, JsCHS, JsCHI and JsFLS were the key genes affecting them. This study will potentially provide more information for light management and polyphenol regulation in walnut cultivation.

Calcium chloride treatment maintains quality and enhances antioxidant capacity of fresh-cut carrots through modulating DcCMLs-DcCAMTA3-mediated phenylpropanoid pathway

Whitening and microbial infection are two major reasons to induce quality deterioration in fresh-cut carrots. Calcium chloride (CaCl2) could enhance the quality attributes in fresh-cut carrots, but the molecular mechanism is still not illuminated. In this study, CaCl2 treatment suppressed the increases of whiteness index and total bacterial count as well as enhanced the antioxidant capacity of fresh-cut carrots. In addition, CaCl2 treatment promoted phenolics accumulation by upregulating the transcripts and enzyme activity of phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H) and 4-coumarate coenzyme A ligase (4CL) in phenylpropanoid pathway. However, this treatment inhibited the increase of enzyme activity and gene expression of cinnamyl alcohol dehydrogenase (CAD), thus suppressing lignin synthesis and whitening process in fresh-cut carrots. Moreover, a calmodulin-binding transcription activator (CAMTA) transcription factor DcCAMTA3, was induced in response to CaCl2 treatment. DcCAMTA3 activated the transcription of DcPAL1, DcC4H and Dc4CL1 genes and repressed the expression of DcCAD1 gene by directly targeting the CG-box. DcCML3/11, a calmodulin-like protein, interacted with DcCAMTA3, and this interaction enhanced DcCAMTA3-mediated transcriptional activation of DcPAL1, DcC4H and Dc4CL1 and repression of DcCAD1. Therefore, CaCl2 treatment is effective in maintaining quality and enhancing antioxidant capacity of fresh-cut carrots by the cooperative action of DcCMLs and DcCAMTA3 in positively regulating phenylpropanoid pathway.

Dietary curcumin supplementation alleviates diquat-induced oxidative stress in the liver of broilers

This study purposed to investigate the alleviating effect of dietary curcumin supplementation on oxidative stress in the liver of broilers induced by diquat. One-day-old Cobb broilers (400) were selected and randomly divided into 5 groups, with 8 replicates and 10 broilers per replicate. The control group and the diquat group were fed the basal diet, while the curcumin supplementation groups were fed the basal diet supplemented with different amounts of curcumin (50, 100, and 150 mg/kg). On d 21 of the test, 1 broiler was randomly selected from each replicate and intraperitoneally injected with 20 mg/mL of diquat solution at a dose of 1 mL/kg BW or equivalent physiological saline (for the control group). After 48 h of feeding, the selected broilers were slaughtered for analysis. The results show that diquat treatment reduced the antioxidant capacity of the liver, caused oxidative stress, and affected its lipid metabolism. However, diet supplementation using curcumin completely or partially reversed the effect of diquat on the liver of broilers. The blood alanine aminotransferase activity, total bilirubin and total protein levels, and liver Caspase-3 mRNA abundance in broilers were lower or significantly lower in the curcumin supplementation group than in the diquat group (P < 0.05). The curcumin supplementation groups had significantly higher total antioxidant capacity activity but significantly lower malondialdehyde in the liver of broilers than the diquat group (P < 0.05). The blood triglyceride level of broilers was lower or significantly lower in the curcumin supplementation groups than in the diquat group (P < 0.05). The activities of cetyl coenzyme A carboxylase in the liver were significantly lower in the 150 mg/kg curcumin supplementation groups than in the DQ group (P < 0.05). In conclusion, dietary curcumin supplementation could ameliorate the effects of diquat-induced oxidative stress on the antioxidant capacity, tissue morphology, and lipid metabolism of the liver of broilers, thus protecting the liver. The recommended dosage for broiler diets is 100 to 150 mg/kg curcumin.

Exogenous methyl jasmonate combined with Ca2+ promote resveratrol biosynthesis and stabilize sprout growth for the production of resveratrol-rich peanut sprouts

Promoting resveratrol accumulation in plants and utilizing resveratrol-rich plants as raw materials for the development of functional foods is a promising development direction. The effects of methyl jasmonate (MeJA), in combination with CaCl2 and Ca2+ inhibitors, on physiological metabolism and resveratrol enrichment of peanut sprouts were investigated. MeJA combined with CaCl2 increased Ca2+ content, calmodulin content, and Ca2+- adenosine triphosphatase activity, as well as upregulated calcium-binding proteinase expression levels. Treatment with MeJA plus CaCl2 significantly increased peroxidase and superoxide dismutase activities and antioxidant capacities, significantly decreased the content of malondialdehyde and hydrogen peroxide, which resulted in a significantly increased in sprout length and fresh weight, and alleviated the inhibition of sprout growth. MeJA plus CaCl2 significantly increased the activities of phenylalanine ammonia-lyase and 4-coumarate coenzyme A ligase and upregulated the expression levels of phenylalanine ammonia-lyase, cinnamic acid 4-hydroxylase, and resveratrol synthase, thus significantly increasing resveratrol content. However, MeJA combined with Ca2+ antagonists reversed these effects. These results indicate that MeJA interacts with Ca2+ to promote resveratrol synthesis in peanut sprouts and to improve sprout stress tolerances.

Study of blood microcirculation in the area of purulent inflammatory process in patients with phlegmon of the maxillofacial region with the analyzer “LASMA PF”

The article is devoted to the assessment of the state of the microcirculatory bed of the skin in the purulent-inflammatory process in patients with phlegmon of the maxillofacial region who were treated in the maxillofacial department of the Moscow Pirogov State Clinical Hospital. Microcirculation parameters were recorded by laser Doppler flowmetry (LDF) with simultaneous temperature recording directly from the skin in the area of purulent-inflammatory infiltrate, by applying the apparatus of the analyzer «LASMA PF» to the patient’s skin. The diagnosis by the analyzer «LAZMA PF» is based on the simultaneous assessment of the activity of the tissue coenzyme of the reduced nicotinamide adenine dinucleotide (NADH) by fluorescence spectroscopy and indicators of microcirculation of blood flow by laser Doppler flowmetry. It is concluded that the differences obtained during the study between microcirculatory disorders in the purulent-inflammatory process and the healthy side indicate the need for drug therapy that regulates blood rheology and the state of the microcirculatory bed.

Ozone mitigates the flesh discoloration in response to 1-methylcyclopropene by promoting anthocyanin biosynthesis in postharvest nectarines

The effects of combined treatments with 1-methylcyclopropene (1-MCP) and ethylene absorbent (EA) or ozone (O3) on the anthocyanins biosynthesis in nectarines stored at 0 °C were investigated during the storage period. The results indicated that 1-MCP+EA and 1-MCP+O3 effectively inhibited respiration and ethylene release. 1-MCP+EA reduced anthocyanins accumulation and inhibited the red coloration in pulp, whereas 1-MCP+O3 accelerated the red pigmentation. Contrasted to the 1-MCP group, 1-MCP+O3 exhibited higher anthocyanins contents, expression levels of PpPAL1, PpCHS, PpDFR, PpANS, PpUFGT, and PpMYB10.2 and activities of 4-coumarate coenzyme A ligase, flavanone 3-hydroxylase, dihydroflavonol 4-reductase, uridine diphosphate flavonoid 3-O-glucosyltransferase involved in the phenylpropanoid pathway. These results indicated that 1-MCP+O3 promoted the anthocyanins biosynthesis due to the up-regulation of enzymes activities and anthocyanins biosynthetic genes and transcription factor. This study may shed new light on improving quality and color development and optimizing 1-MCP application for postharvest preservation of fruit.

Characteristics of digested sludge-derived biochar for promoting methane production during anaerobic digestion of waste activated sludge

This study investigated a novel method for enhancing methane production during anaerobic digestion of waste activated sludge with digested sludge-derived biochar (DSBC). Using response surface methodology, the following process conditions for DSBC synthesis were optimized: heating rate = 13.23 °C/min, pyrolysis temperature = 516 °C, and heating time = 192 min. DSBC significantly enhanced the methane production by 48 % and improved key coenzyme activity that accelerated the bioconversion of organic matter while promoting the decomposition and transformation of volatile fatty acids. Consequently, the lag period of methane production was shortened to 4.89 days, while the average proportion of methane greatly increased to 73.22%. Thus, DSBC could facilitate efficient methanogenesis in the anaerobic system by promoting electron transfer between syntrophic partners through the charge-discharge cycle of surface oxygen-containing functional groups. The study provides a reference for the resource utilization of anaerobic sludge residues and efficient anaerobic methanogenesis from sludge.

Effects of Exogenous Nitric Oxide Treatment on Grape Berries Against Botrytis cinerea and Alternaria alternata Related Enzymes and Metabolites.

Postharvest losses of grape berries caused by the pathogenic fungi Botrytis cinerea and Alternaria alternata have been widely reported, and nitric oxide (NO) as a plant signaling molecule to control postharvest diseases has recently become an active research topic. This study aimed to investigate the regulatory effect of NO on the interaction between grape berries and fungi. During interactions between grape berries and pathogenic fungi, treatment with 10 mM sodium nitroprusside (SNP, an NO donor) delayed the decline of the physiological quality of the grape berries and had positive effects on the weight loss rate, firmness, and respiration intensity. SNP treatment increased the activities of superoxide dismutase (SOD) and polyphenol oxidase (PPO) and inhibited the activities of peroxidase (POD) and catalase (CAT) of grape berries during the resistance to fungal pathogen infection. In addition, the increase in browning degree and the accumulation of hydrogen peroxide were inhibited by SNP treatment. In the phenylpropane metabolic pathway, the activities of phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), and 4-coumaric acid coenzyme A ligase (4CL) were increased during the activation of grape berries during the resistance to pathogen infection by SNP, and the intermediate metabolites lignin, flavonoids, and total phenols were accumulated. In addition, SNP treatment had a regulatory effect on the gene expression levels of SOD, POD, PPO, PAL, and 4CL. These results suggested that SNP treatment was effective for the preservation and disease reduction of grape berries.

A concerted mechanism involving ACAT and SREBPs by which oxysterols deplete accessible cholesterol to restrict microbial infection.

Most of the cholesterol in the plasma membranes (PMs) of animal cells is sequestered through interactions with phospholipids and transmembrane domains of proteins. However, as cholesterol concentration rises above the PM's sequestration capacity, a new pool of cholesterol, called accessible cholesterol, emerges. The transport of accessible cholesterol between the PM and the endoplasmic reticulum (ER) is critical to maintain cholesterol homeostasis. This pathway has also been implicated in the suppression of both bacterial and viral pathogens by immunomodulatory oxysterols. Here, we describe a mechanism of depletion of accessible cholesterol from PMs by the oxysterol 25-hydroxycholesterol (25HC). We show that 25HC-mediated activation of acyl coenzyme A: cholesterol acyltransferase (ACAT) in the ER creates an imbalance in the equilibrium distribution of accessible cholesterol between the ER and PM. This imbalance triggers the rapid internalization of accessible cholesterol from the PM, and this depletion is sustained for long periods of time through 25HC-mediated suppression of SREBPs and continued activation of ACAT. In support of a physiological role for this mechanism, 25HC failed to suppress Zika virus and human coronavirus infection in ACAT-deficient cells, and Listeria monocytogenes infection in ACAT-deficient cells and mice. We propose that selective depletion of accessible PM cholesterol triggered by ACAT activation and sustained through SREBP suppression underpins the immunological activities of 25HC and a functionally related class of oxysterols.

Evaluation of the synergistic effects of biochar and biogas residue on CO2 and CH4 emission, functional genes, and enzyme activity during straw composting

This study examined the effects of biochar, biogas residue, and their combined amendments on CO2 and CH4 emission, enzyme activity, and related functional genes during rice straw composting. Results showed that the biogas residue increased CO2 and CH4 emissions by 13.07 % and 74.65 %, while biochar had more obvious inhibition. Biogas residue addition enhanced functional gene abundance more than biochar. Biogas residue raised the methanogens mcrA gene by 2.5 times. Biochar improved the Acetyl-CoA synthase and β-glucosidase activities related to carbon fixation and decreased coenzyme activities related to methanogens. Biochar and biogas residue combined amendments enhanced the acsB gene abundance for CO2 assimilation process and decreased methyl-coenzyme M reductase α subunit activity. Pearson correlation analysis indicated that organic matter was the significant variable affecting CO2 and CH4 emissions (P < 0.01). These results indicated biochar played significant roles in carbon loss and greenhouse emissions caused by biogas residue incorporation during composting.

Stimulating Effect of Trichococcus flocculiformis on a Coculture of Syntrophomonas wolfei and Methanospirillum hungatei.

ABSTRACTSyntrophic anaerobic consortia comprised of fatty acid-degrading bacteria and hydrogen/formate-scavenging methanogenic archaea are of central importance for balanced and resilient natural and manufactured ecosystems: anoxic sediments, soils, and wastewater treatment bioreactors. Previously published studies investigated interaction between the syntrophic bi-cultures, but little information is available on the influence of fermentative bacteria on syntrophic fatty acid oxidation, even though fermentative organisms are always present together with syntrophic partners in the above-mentioned ecosystems. Here, we present experimental observations of stimulated butyrate oxidation and methane generation by a coculture of Syntrophomonas wolfei with any of the following methanogens: Methanospirillum hungatei, Methanobrevibacter arboriphilus, or Methanobacterium formicicum due to the addition of a fermentative Trichococcus flocculiformis strain ES5. The addition of T. flocculiformis ES5 to the syntrophic cocultures led to an increase in the rates of butyrate consumption (120%) and volumetric methane production (150%). Scanning electron microscopy of the most positively affected coculture (S. wolfei, M. hungatei, and T. flocculiformis ES5) revealed a tendency of T. flocculiformis ES5 to aggregate with the syntrophic partners. Analysis of coculture’s proteome with or without addition of the fermentative bacterium points to a potential link with signal transducing systems of M. hungatei, as well as activation of additional butyryl coenzyme A dehydrogenase and an electron transfer flavoprotein in S. wolfei.IMPORTANCE Results from the present study open doors to fascinating research on complex microbial cultures in anaerobic environments (of biotechnological and ecological relevance). Such studies of defined mixed populations are critical to understanding the highly intertwined natural and engineered microbial systems and to developing more reliable and trustable metabolic models. By investigating the existing cultured microbial consortia, like the ones described here, we can acquire knowledge on microbial interactions that go beyond “who feeds whom” relations but yet benefit the parties involved. Transfer of signaling compounds and stimulation of gene expression are examples of indirect influence that members of mixed communities can exert on each other. Understanding such microbial relationships will enable development of new sustainable biotechnologies with mixed microbial cocultures and contribute to the general understanding of the complex natural microbial interactions.

Exogenous melatonin stimulated isoflavone biosynthesis in NaCl-stressed germinating soybean (Glycine max L.)

Melatonin (MT) has gained increasing attention due to its pleiotropic effects. In this study, the function of exogenous MT on the response to NaCl stress and isoflavone biosynthesis in germinating soybeans was investigated. Results showed the exogenous MT (100 μM) application neutralised the negative effects of NaCl stress (60 mM), induced sprout growth, biomass and fluorescence intensity of intracellular free calcium, decreased malondialdehyde, H2O2 content and fluorescence intensity of O2•-, and enhanced superoxide dismutase, catalase and peroxidas activities of germinating soybeans. Meanwhile, total flavonoids and different forms of isoflavone content were enhanced by MT application, not only companied by the up-regulated relative gene expression of cinnamic acid 4-hydroxylase chalcone reductase, chalcone isomerase 1A, isoflavone reductase and isoflavone synthase 1 that involved in isoflavone biosynthesis, but also increased activities of phenylalanine ammonia lyase and 4-coumarate coenzyme A ligase. Given the evidence from the present study, it's proposed that the exogenous MT could relieve NaCl stress and stimulate isoflavone biosynthesis in germinating soybeans.

Molecular Manipulation of the miR396 and miR399 Expression Modules Alters the Response of Arabidopsis thaliana to Phosphate Stress.

In plant cells, the molecular and metabolic processes of nucleic acid synthesis, phospholipid production, coenzyme activation and the generation of the vast amount of chemical energy required to drive these processes relies on an adequate supply of the essential macronutrient, phosphorous (P). The requirement of an appropriate level of P in plant cells is evidenced by the intricately linked molecular mechanisms of P sensing, signaling and transport. One such mechanism is the posttranscriptional regulation of the P response pathway by the highly conserved plant microRNA (miRNA), miR399. In addition to miR399, numerous other plant miRNAs are also required to respond to environmental stress, including miR396. Here, we exposed Arabidopsis thaliana (Arabidopsis) transformant lines which harbor molecular modifications to the miR396 and miR399 expression modules to phosphate (PO4) starvation. We show that molecular alteration of either miR396 or miR399 abundance afforded the Arabidopsis transformant lines different degrees of tolerance to PO4 starvation. Furthermore, RT-qPCR assessment of PO4-starved miR396 and miR399 transformants revealed that the tolerance displayed by these plant lines to this form of abiotic stress most likely stemmed from the altered expression of the target genes of these two miRNAs. Therefore, this study forms an early step towards the future development of molecularly modified plant lines which possess a degree of tolerance to growth in a PO4 deficient environment.

Enhanced Biosynthesis of Fatty Acids Is Associated with the Acquisition of Ciprofloxacin Resistance in Edwardsiella tarda.

ABSTRACTMisuse and overuse of antibiotics drive the selection and spread of antibiotic-resistant bacteria. Although genetic mutations have been well defined for different types of antibiotic resistance, ways to revert antibiotic resistance are largely unexplored. Here, we adopted a proteomics approach to investigate the mechanism underlying ciprofloxacin resistance in Edwardsiella tarda, a representative pathogen that infects both economic animal species and human beings. By comparing the protein expression profiles of ciprofloxacin-sensitive and -resistant E. tarda, a total of 233 proteins of differential abundance were identified, where 53 proteins belong to the functional categories of metabolism, featuring a disrupted pyruvate cycle and decreased energy metabolism but increased fatty acid biosynthesis. The altered pyruvate cycle and energy metabolism were confirmed by gene expression and biochemical assays. Furthermore, the role of fatty acid biosynthesis and quinolone resistance were explored. The expression level and enzymatic activity of acetyl coenzyme A (acetyl-CoA) carboxylase, the first step of fatty acid biosynthesis, were increased in ciprofloxacin-resistant E. tarda. Treatment of ciprofloxacin-resistant E. tarda with acetyl-CoA carboxylase and 3-oxoacyl-[acyl carrier protein] synthase II inhibitors, 2-aminooxazole and triclosan, respectively, reduced the expression of fatty acid biosynthesis and promoted quinolone-mediated killing efficacy to antibiotic-resistant bacteria. Similar results were obtained in clinically isolated E. tarda strains. Our study suggests that energy metabolism has been reprogramed in ciprofloxacin-resistant bacteria that favor the biosynthesis of fatty acid, presenting a novel target to tackle antibiotic-resistant bacteria.IMPORTANCE Edwardsiella tarda is the causative agent of edwardsiellosis, which imposes huge challenges on clinics and aquaculture. Due to the overuse of antibiotics, the emergence and spread of antibiotic-resistant E. tarda threaten human health and animal farming. However, the mechanism of ciprofloxacin resistance in E. tarda is still lacking. Here, iTRAQ (isobaric tags for relative and absolute quantification)-based proteomics was performed to identify a differential proteome between ciprofloxacin-sensitive and -resistant E. tarda. The fluctuated pyruvate cycle and reduced energy metabolism and elevated fatty acid biosynthesis are metabolic signatures of ciprofloxacin resistance. Moreover, inhibition of biosynthesis of fatty acids promotes quinolone-mediated killing efficacy in both lab-evolved and clinically isolated strains. This study reveals that a ciprofloxacin resistance mechanism is mediated by the elevated biosynthesis of fatty acids and the depressed pyruvate metabolism and energy metabolism in E. tarda. These findings provide a novel understanding for the ciprofloxacin resistance mechanism in E. tarda.

Lipoic acid. Kinetics and pluripotent biological properties and derivatives.

Lipoic acid (LA) is globally known and its supplements are widely used. Despite its importance for the organism it is not considered a vitamin any more. The multiple metabolic forms and the differences in kinetics (absorption, distribution and excretion), as well as the actions of its enantiomers are analysed in the present article together with its biosynthetic path. The proteins involved in the transfer, biotransformation and activity of LA are mentioned. Furthermore, the safety and the toxicological profile of the compound are commented, together with its stability issues. Mechanisms of lipoic acid intervention in the human body are analysed considering the antioxidant and non-antioxidant characteristics of the compound. The chelating properties, the regenerative ability of other antioxidants, the co-enzyme activity and the signal transduction by the implication in various pathways will be discussed in order to be elucidated the pleiotropic effects of LA. Finally, lipoic acid integrating analogues are mentioned under the scope of the multiple pharmacological actions they acquire towards degenerative conditions.