Plant-soil interactions alter nitrogen and phosphorus dynamics in an advancing subarctic treeline.

Treelines advance due to climate warming. The impacts of this vegetation shift on plant-soil nutrient cycling are still uncertain, yet highly relevant as nutrient availability stimulates tree growth. Here, we investigated nitrogen (N) and phosphorus (P) in plant and soil pools along two tundra-forest transects on Kola Peninsula, Russia, with a documented elevation shift of birch-dominated treeline by 70 m during the last 50 years. Results show that although total N and P stocks in the soil-plant system did not change with elevation, their distribution was significantly altered. With the transition from high-elevation tundra to low-elevation forest, P stocks in stones decreased, possibly reflecting enhanced weathering. In contrast, N and P stocks in plant biomass approximately tripled and available P and N in the soil increased fivefold toward the forest. This was paralleled by decreasing carbon (C)-to-nutrient ratios in foliage and litter, smaller C:N:P ratios in microbial biomass, and lower enzymatic activities related to N and P acquisition in forest soils. An incubation experiment further demonstrated manifold higher N and P net mineralization rates in litter and soil in forest compared to tundra, likely due to smaller C:N:P ratios in decomposing organic matter. Overall, our results show that forest expansion increases the mobilization of available nutrients through enhanced weathering and positive plant-soil feedback, with nutrient-rich forest litter releasing greater amounts of N and P upon decomposition. While the low N and P availability in tundra may retard treeline advances, its improvement toward the forest likely promotes tree growth and forest development.

Enhancing Bromelain Recovery from Pineapple By-Products: A Sustainable Approach for Value Addition and Waste Reduction.

Pineapple by-products are good sources of bromelain, a complex enzyme with commercial applications. This study evaluated the feasibility of producing bromelain powders from pineapple waste using an organic solvent-free approach. Pineapple by-products (from var. MD2), including cores, peels, crowns, stems, and basal stems, were homogenized with deionized water, and the pH of the mixture was adjusted to 4.5 and 9 (isoelectric points of fruit bromelain and stem bromelain), clarified, ultra-filtered, and freeze-dried to produce bromelain powders. The enzymatic activity of the bromelain powders was measured using the gelatin and casein digestion methods. The bromelain powders from the crowns did not show significant enzymatic activity (p < 0.05). Meanwhile, bromelain powders produced from the cores and peels had an enzymatic activity of 694 gelatin digesting units (GDU)/g and 124 casein digesting units (CDU)/mg, and 1179 GDU/g and 217 CDU/mg, respectively. Bromelain powders from the basal stems showed the highest enzymatic activity (2909 GDU/g and 717 CDU/mg). Increasing the pH of the liquid mixture before the purification and freeze drying significantly (p < 0.05) reduced the enzymatic activity of the bromelain powders. Using a practical and organic solvent-free approach, this study demonstrates the feasibility of producing bromelain powders with high enzymatic activity from pineapple waste.

Antifungal activity of copper oxide nanoparticles derived from Zizyphus spina leaf extract against Fusarium root rot disease in tomato plants.

Incorporating green chemistry concepts into nanotechnology is an important focus area in nanoscience. The demand for green metal oxide nanoparticle production has grown in recent years. The beneficial effects of using nanoparticles in agriculture have already been established. Here, we highlight some potential antifungal properties of Zizyphus spina leaf extract-derived copper oxide nanoparticles (CuO-Zs-NPs), produced with a spherical shape and defined a 13-30 nm particle size. Three different dosages of CuO-Zs-NPs were utilized and showed promising antifungal efficacy in vitro and in vivo against the selected fungal strain of F. solani causes tomato root rot disease, which was molecularly identified with accession number (OP824846). In vivo  results indicated that, for all CuO-Zs-NPs concentrations, a significant reduction in Fusarium root rot disease occurred between 72.0 to 88.6% compared to 80.5% disease severity in the infected control. Although treatments with either the chemical fungicide (Kocide 2000) showed a better disease reduction and incidence with (18.33% and 6.67%) values, respectively, than CuO-Zs-NPs at conc. 50 mg/l, however CuO-Zs-NPs at 250 mg/l conc. showed the highest disease reduction (9.17 ± 2.89%) and lowest disease incidence (4.17 ± 3.80%). On the other hand, CuO-Zs-NPs at varied values elevated the beneficial effects of tomato seedling vigor at the initial stages and plant growth development compared to either treatment with the commercial fungicide or Trichoderma Biocide. Additionally, CuO-Zs-NPs treatments introduced beneficial results for tomato seedling development, with a significant increase in chlorophyll pigments and enzymatic activity for CuO-Zs-NPs treatments. Additionally, treatment with low concentrations of CuO-Zs-NPs led to a rise in the number of mature pollen grains compared to the immature ones.  however the data showed that CuO-Zs-NPs have a unique antifungal mechanism against F. solani, they  subsequently imply that CuO-Zs-NPs might be a useful environmentally friendly controlling agent for the Fusarium root rot disease that affects tomato plants.

Antioxidant potentialities and gastroprotective effect of Reichardia picroides extracts on Ethanol/HCl induced gastric ulcer rats.

The goal of this study was to determine for the first time the polyphenol content, antioxidant, and gastroprotective properties of the roots and leaves of Reichardia picroides. TPC considerably varied as a function of organs and solvent nature and ranged from 50 to 284.80 mg GAE/g DW. Leaves exhibited the highest amount of phenolics by using acetone 70%, the same tendency was observed for antioxidant activity. Besides, in vivo gastro-protective effects following HCl/EtOH-induced ulcer models displayed that roots extract at a high dose (500 mg) seemed to be the best performing extract with a decrease of ulceration index (UI) and an increase in the percentage of protection (PP), SOD, CAT, and GPX activities. All these data have been proved with principal component analysis (PCA). Overall, the results indicated that R. picroides could be considered a valuable source of natural compounds, which are beneficial for human health.

Intercropping system modulated soil-microbe interactions that enhanced the growth and quality of flue-cured tobacco by improving rhizospheric soil nutrients, microbial structure, and enzymatic activities.

As the promotive/complementary mechanism of the microbe-soil-tobacco (Nicotiana tabacum L.) interaction remains unclear and the contribution of this triple interaction to tobacco growth is not predictable, the effects of intercropping on soil nutrients, enzymatic activity, microbial community composition, plant growth, and plant quality were studied, and the regulatory mechanism of intercropping on plant productivity and soil microenvironment (fertility and microorganisms) were evaluated. The results showed that the soil organic matter (OM), available nitrogen (AN), available phosphorus (AP), available potassium (AK), the urease activity (UE) and sucrase activity (SC), the diversity, abundance, and total and unique operational taxonomic units (OTUs) of bacteria and fungi as well as plant biomass in T1 (intercropping onion), T2 (intercropping endive), and T3 (intercropping lettuce) treatments were significantly higher than those of the controls (monocropping tobacco). Although the dominant bacteria and fungi at the phylum level were the same for each treatment, LEfSe analysis showed that significant differences in community structure composition and the distribution proportion of each dominant community were different. Proteobacteria, Acidobacteria, and Firmicutes of bacteria and Ascomycota and Basidiomycetes of fungi in T1, T2, and T3 treatments were higher than those of the controls. Redundancy analysis (RDA) suggested a close relation between soil characteristic parameters and microbial taxa. The correlation analysis between the soil characteristic parameters and the plant showed that the plant biomass was closely related to soil characteristic parameters. In conclusion, the flue-cured tobacco intercropping not only increased plant biomass and improved chemical quality but also significantly increased rhizospheric soil nutrient and enzymatic activities, optimizing the microbial community composition and diversity of rhizosphere soil. The current study highlighted the importance of microbe-soil-tobacco interactions in maintaining plant productivity and provided the potential fertilization practices in flue-cured tobacco production to maintain ecological sustainability.

[Seasonal Regulation of Soil Microbial Carbon and Phosphorus Metabolisms in an Apple Orchard: Evidence from the Enzymatic Stoichiometry Method].

Soil microbial carbon (C), nitrogen (N), and phosphorus (P) nutrient requirements and metabolic limitations are closely related to the availability of environmental nutrients. However, it is unclear how manure and chemical fertilization shift nutrient limitations for microbes in terms of the soil enzymatic stoichiometry in an apple orchard. Therefore, based on the long-term experiment located in an apple orchard established in 2008, this study applied the theory and method of soil enzyme stoichiometry to systematically investigate the effects of the combined application of manure and chemical fertilizers on soil C, N, and P turnover-related enzyme activities (ß-1,4-glucosidase, BG; leucine aminopeptidase, NAG; ß-1,4-N-acetylglucosaminidase, LAP; and acid or alkaline phosphatase, PHOS) and their stoichiometric characteristics and analyzed their relationships with environmental factors and microbial carbon use efficiency. The experiment was designed with four treatments, such as, no fertilization input as the control (CK), single application of chemical fertilizer (NPK), combined application of manure and chemical fertilizer (MNPK), and single application of manure (M). The results revealed that:① at different growth stages of fruit trees, the soil microbial biomass C (microC) content of manure fertilizer treatments (MNPK and M) was significantly higher than that of no manure fertilizer treatments (CK and NPK). The content of microbial biomass N (microN) in the NPK, MNPK, and M treatments increased by 89%, 269%, and 213%, respectively, compared with that in CK (P<0.05). ② Compared with those in the fertilization treatments, CK had higher leaf N and P contents (29.8 g·kg-1 and 2.17 g·kg-1) at the germination stage, and the leaf P content at the germination stage alone was significantly negatively correlated with soil available phosphorus (AP) content. ③ Soil enzyme stoichiometry analysis demonstrated that all data points in this study were above the 1:1 line, indicating that microbial communities had a strong phosphorus limitation. The range of vector length and angle was 0.56-0.79 and 59.3°-67.7°, respectively, in the growth period of fruit trees, and the vector angle was >45° in this study, which also reflected the strong phosphorus limitation of microorganisms. ④ RDA and random forest model analysis showed that organic carbon and available nitrogen (AN) were the main physical and chemical factors affecting vector length; AP, AN, and soil water content were the main physical and chemical factors affecting vector angle. Combined with SEM analysis, AN and dissolved organic carbon (DOC) directly affected microC and microN, AP directly affected microP and microN, DOC and AP directly affected vector length, and AP and microN directly affected vector angle. In addition, microbial carbon utilization was positively correlated with vector length and negatively correlated with vector angle. In summary, the combined application of manure and chemical fertilizers regulated microbial carbon and phosphorus metabolism by affecting soil carbon and phosphorus content at different growth stages of fruit trees, thereby affecting microbial carbon utilization. This study provides a scientific basis for manure and chemical fertilizers to improve soil quality and maintain soil health.

Antimicrobial activity and enzymatic analysis of endophytes isolated from Codonopsis pilosula.

The roots of the medicinal plant Codonopsis pilosula (Franch.) Nannf (C. pilosula) possess most medicinal supplements. In current research on C. pilosula root endophytes were isolated, identified, and evaluated for their antimicrobial activity against human pathogens such as Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Salmonella typhi, and Pseudomonas aeruginosa and the fungi Candida albicans and Aspergillus niger. Endophytes C.P-8 and C.P-20 exhibited very significant antimicrobial activity, the secondary metabolite of C.P-8 registered at retention time 24.075 by HPLC analysis. Significant minimum inhibitory concentration (MIC) of C.P-8 was exhibited at 250 µg/ml against S. aureus and 500 µg/ml against B. subtilis. Qualitative, quantitative analyses, and partial purification of enzymes and purity was analysed by molecular weight determined by SDS‒PAGE of enzymes produced by C.P-20, amylase-64 kDa, protease-64 kDa, chitinase-30 kDa, and cellulase-54 kDa. Optimum pH and temperature of the partially purified enzymes, was carried out. The partially purified enzymes from C.P-20 displayed maximum activity at pH 6-7 and temperatures of 40°C-45°C. Moreover, the above endophytes will be useful tools for producing active enzymes and active bioantimicrobial agents against human pathogens.

Mineral-Solubilizing Bacteria-Mediated Enzymatic Regulation and Nutrient Acquisition Benefit Cotton's (Gossypium hirsutum L.) Vegetative and Reproductive Growth.

Many farmers' incomes in developing countries depend on the cultivation of major crops grown in arid and semi-arid regions. The agricultural productivity of arid and semi-arid areas primarily relies on chemical fertilizers. The effectiveness of chemical fertilizers needs to improve by integration with other sources of nutrients. Plant growth-promoting bacteria can solubilize nutrients, increase plant nutrient uptake, and supplement chemical fertilizers. A pot experiment evaluated the promising plant growth-promoting bacterial strain's effectiveness in promoting cotton growth, antioxidant enzymes, yield, and nutrient uptake. Two phosphate solubilizing bacterial strains (Bacillus subtilis IA6 and Paenibacillus polymyxa IA7) and two zinc solubilizing bacterial strains (Bacillus sp. IA7 and Bacillus aryabhattai IA20) were coated on cotton seeds in a single as well as co-inoculation treatments. These treatments were compared with uninoculated controls in the presence and absence of recommended chemical fertilizer doses. The results showed the co-inoculation combination of Paenibacillus polymyxa IA7 and Bacillus aryabhattai IA20 significantly increased the number of bolls, seed cotton yield, lint yield, and antioxidants activities, including superoxide dismutase, guaiacol peroxidase, catalase, and peroxidase. Co-inoculation combination of Bacillus subtilis IA6 and Bacillus sp. IA16 promoted growth attributes, including shoot length, root length, shoot fresh weight, and root fresh weight. This co-inoculation combination also increased soil nutrient content. At the same time, Paenibacillus polymyxa IA7 + Bacillus aryabhattai IA20 increased nutrient uptake by plant shoots and roots compared.

Antioxidant, Tyrosinase, α-Glucosidase, and Elastase Enzyme Inhibition Activities of Optimized Unripe Ajwa Date Pulp (Phoenix dactylifera) Extracts by Response Surface Methodology.

The Ajwa date (Phoenix dactylifera L., Arecaceae family) is a popular edible fruit consumed all over the world. The profiling of the polyphenolic compounds of optimized unripe Ajwa date pulp (URADP) extracts is scarce. The aim of this study was to extract polyphenols from URADP as effectively as possible by using response surface methodology (RSM). A central composite design (CCD) was used to optimize the extraction conditions with respect to ethanol concentration, extraction time, and temperature and to achieve the maximum amount of polyphenolic compounds. High-resolution mass spectrometry was used to identify the URADP's polyphenolic compounds. The DPPH-, ABTS-radical scavenging, α-glucosidase, elastase and tyrosinase enzyme inhibition of optimized extracts of URADP was also evaluated. According to RSM, the highest amounts of TPC (24.25 ± 1.02 mgGAE/g) and TFC (23.98 ± 0.65 mgCAE/g) were obtained at 52% ethanol, 81 min time, and 63 °C. Seventy (70) secondary metabolites, including phenolic, flavonoids, fatty acids, and sugar, were discovered using high-resolution mass spectrometry. In addition, twelve (12) new phytoconstituents were identified for the first time in this plant. Optimized URADP extract showed inhibition of DPPH-radical (IC50 = 87.56 mg/mL), ABTS-radical (IC50 = 172.36 mg/mL), α-glucosidase (IC50 = 221.59 mg/mL), elastase (IC50 = 372.25 mg/mL) and tyrosinase (IC50 = 59.53 mg/mL) enzymes. The results revealed a significant amount of phytoconstituents, making it an excellent contender for the pharmaceutical and food industries.

Microalgae biomass as a renewable biostimulant: meat processing industry effluent treatment, soil health improvement, and plant growth.

Microalgae biomass contributes to effluent bioremediation. It is a concentrated source of nutrients and organic carbon, making it a potential alternative as a soil biostimulant. In this context, this study aimed to evaluate the soil application of microalgae biomass produced from the meat processing industry effluent treatment. The biomass was applied dry and as a mixture to demonstrate its potential to increase plant production and soil metabolic functions, analyzed short-term. Doses of 0.25%, 0.5%, 1%, and 2% biomass were applied in soils from (i) Horizon A: taken at a depth between 0 and 10 cm and; (ii) Horizon B: taken at a depth between 20 and 40 cm. Corn growth (Zea Mays L.), basal soil respiration, microbial biomass carbon, total organic carbon, ß-glucosidase, acid phosphatase, arylsulfatase, and urease enzymatic activity were evaluated in each sample. It is concluded that applying 2% microalgae biomass led to higher basal soil respiration, microbial biomass carbon, and ß-glucosidase, acid phosphatase, arylsulfatase enzymatic activity in both soils. On the other hand, boron may have contributed to urease activity reduction in Soil A. Although 2% biomass led to higher soils characteristics, that dose did not promote higher plant growth. Hence, considering that plant growth must be in line with changes in soil characteristics, the result that provided the higher plant shoot dry matter mass was by applying 0.55% biomass in both soils. Therefore, the application of microalgae biomass produced from a meat processing industry effluent treatment promoted a biologically active soil and boosted plant growth.

Gene coexpression networks allow the discovery of two strictosidine synthases underlying monoterpene indole alkaloid biosynthesis in Uncaria rhynchophylla.

Plant-derived monoterpene indole alkaloids (MIAs) from Uncaria rhynchophylla (UR) have huge medicinal properties in treating Alzheimer's disease, Parkinson's disease, and depression. Although many bioactive UR-MIA products have been isolated as drugs, their biosynthetic pathway remains largely unexplored. In this study, untargeted metabolome identified 79 MIA features in UR tissues (leaf, branch stem, hook stem, and stem), of which 30 MIAs were differentially accumulated among different tissues. Short time series expression analysis captured 58 pathway genes and 12 hub regulators responsible for UR-MIA biosynthesis and regulation, which were strong links with main UR-MIA features. Coexpression networks further pointed to two strictosidine synthases (UrSTR1/5) that were coregulated with multiple MIA-related genes and highly correlated with UR-MIA features (r > 0.7, P < 0.005). Both UrSTR1/5 catalyzed the formation of strictosidine with tryptamine and secologanin as substrates, highlighting the importance of key residues (UrSTR1: Glu309, Tyr155; UrSTR5: Glu295, Tyr141). Further, overexpression of UrSTR1/5 in UR hairy roots constitutively increased the biosynthesis of bioactive UR-MIAs (rhynchophylline, isorhynchophylline, corynoxeine, etc), whereas RNAi of UrSTR1/5 significantly decreased UR-MIA biosynthesis. Collectively, our work not only provides candidates for reconstituting the biosynthesis of bioactive UR-MIAs in heterologous hosts but also highlights a powerful strategy for mining natural product biosynthesis in medicinal plants.

Near-Infrared-Enhanced Dual Enzyme-Mimicking Ag-TiO2-x@Alginate Microspheres with Antibactericidal and Oxygeneration Abilities to Treat Periodontitis.

The effective treatment for periodontitis is to completely and sustainedly eradicate the bacterial pathogens from the complex periodontal pockets. Local sustained-release antibiotics as a complementary treatment after scaling and root planning can sustainedly combat bacterial pathogens in the periodontal pockets to help treat the disease, but the increasing concern of bacterial resistance limits its future use. Here, we reported a local antibacterial system based on microsized multifunctional Ag-TiO2-x encapsulated in alginate (ATA) microspheres. We confirmed that ATA displayed strong photothermally enhanced dual enzyme-mimicking (peroxidase-like and catalase-like) activities and weak photocatalytic activity under 808 nm near-infrared (NIR) irradiation, which could boost the generation of reactive oxygen species (ROS) and O2 in the presence of low-level H2O2. As a result, the ATA/H2O2/NIR system exhibited efficient antibacterial activity against Porphyromonas gingivalis and Streptococcus gordonii in both planktonic and biofilm forms. With the help of ROS, ATA could release Ag+ in concentrations sufficient to inhibit periodontal pathogens as well. Moreover, the in situ-generated oxygen was supposed to alleviate the local hypoxic environment and would help downregulate the lipopolysaccharide-mediated inflammatory response of periodontal stem cells. The in vivo rat periodontitis treatment results demonstrated that the ATA/H2O2/NIR system reduced the bacterial load, relieved inflammation, and improved tissue healing. Our work developed a new local prolonged bactericidal and oxygenation system for enhanced periodontitis. Avoiding the usage of antibiotics and nanomaterials, this strategy showed great promise in adjunctive periodontitis treatment and also in other biomedical applications.

Eco-Friendly Solution Based on Rosmarinus officinalis Hydro-Alcoholic Extract to Prevent Biodeterioration of Cultural Heritage Objects and Buildings.

Biodeterioration of cultural heritage is caused by different organisms capable of inducing complex alteration processes. The present study aimed to evaluate the efficiency of Rosmarinus officinalis hydro-alcoholic extract to inhibit the growth of deteriogenic microbial strains. For this, the physico-chemical characterization of the vegetal extract by UHPLC-MS/MS, its antimicrobial and antibiofilm activity on a representative number of biodeteriogenic microbial strains, as well as the antioxidant activity determined by DPPH, CUPRAC, FRAP, TEAC methods, were performed. The extract had a total phenol content of 15.62 ± 0.97 mg GAE/mL of which approximately 8.53% were flavonoids. The polyphenolic profile included carnosic acid, carnosol, rosmarinic acid and hesperidin as major components. The extract exhibited good and wide spectrum antimicrobial activity, with low MIC (minimal inhibitory concentration) values against fungal strains such as Aspergillus clavatus (MIC = 1.2 mg/mL) and bacterial strains such as Arthrobacter globiformis (MIC = 0.78 mg/mL) or Bacillus cereus (MIC = 1.56 mg/mL). The rosemary extract inhibited the adherence capacity to the inert substrate of Penicillium chrysogenum strains isolated from wooden objects or textiles and B. thuringiensis strains. A potential mechanism of R. officinalis antimicrobial activity could be represented by the release of nitric oxide (NO), a universal signalling molecule for stress management. Moreover, the treatment of microbial cultures with subinhibitory concentrations has modulated the production of microbial enzymes and organic acids involved in biodeterioration, with the effect depending on the studied microbial strain, isolation source and the tested soluble factor. This paper reports for the first time the potential of R. officinalis hydro-alcoholic extract for the development of eco-friendly solutions dedicated to the conservation/safeguarding of tangible cultural heritage.

An extracellular Cu/Zn superoxide dismutase from Neocaridina denticulata sinensis: cDNA cloning, mRNA expression and characterizations of recombinant protein.

Neocaridina denticulata sinensis possesses characters of rapid growth, tenacious vitality, short growth cycle, transparent, and easy feeding. Therefore, it is gradually being developed into an animal model for basic research on decapod crustaceans. Herein, a Cu/Zn superoxide dismutase (Cu/Zn-SOD), named as Nd-ecCu/Zn-SOD, was identified and characterized from N. denticulata sinensis. The full-length cDNA sequence of Nd-ecCu/Zn-SOD is 829 bp containing a 684 bp open reading frame, which encodes a protein of 227 amino acid residues with a typical Sod_Cu domain. The quantitative real-time PCR analysis showed that Nd-ecCu/Zn-SOD mRNA was expressed in all the tested tissues. Under challenge with copper, the mRNA expression of Nd-ecCu/Zn-SOD reached the maximum at 6 h, and decreased until 24 h. After 24 h of exposure, its expression was up-regulated significantly at 36 h. After then its expression sharply decreased with a comeback at 48 h. The result indicated that Nd-ecCu/Zn-SOD might play an important role in the stress response of N. denticulata sinensis. The expression of Nd-ecCu/Zn-SOD in gills challenged with Vibrio parahaemolyticus changed in a time-dependent manner. Nd-ecCu/Zn-SOD was lowly expressed in early developmental stages by RNA-Seq technology, yet it showed that a cyclical rise and fall occurred between middle stages and late stages. In addition, Nd-ecCu/Zn-SOD was recombinantly expressed using E. coli and the recombinant protein was purified as a single band on SDS-PAGE. The recombinant Nd-ecCu/Zn-SOD (rNd-ecCu/Zn-SOD) existed enzymatic activity under a wide range of temperature and pH. The exposure of metal ions was found that Zn2+, Mg2+, Ca2+, Ba2+, and Cu2+ could inhibit the enzymatic activity of rNd-ecCu/Zn-SOD, and Mn2+ increased the enzymatic activity of rNd-ecCu/Zn-SOD. These results indicate that Nd-ecCu/Zn-SOD may play a pivotal role in resistant against oxidative damage and act as a biomarker under stressful environment.

Effective treatment of resistant opportunistic fungi associated with immuno-compromised individuals using silver biosynthesized nanoparticles.

Drug resistance in filamentous fungus to antifungal medicines is a huge problem in biomedical applications; so, an effective strategy for treating opportunistic fungal infections is needed. Mentha piperita is a very fascinating plant to treat a variety of ailments as home remedies. Eighteen strains of Aspergillus species were used for this study which are having a unique antifungal resistance profile in presence of silver nanoparticles (AgNPs). AgNPs were prepared, using an aqueous extract of M. Piperita and characterized it by various techniques. Structural properties of AgNPs were systematically studied using X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM), Fourier-transform infrared spectroscopy (FT-IR), and Raman measurement, which emanate the single-phase fcc structure of silver nanoparticles. The spherical nature and elemental analysis of as-synthesized AgNPs were confirmed using scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) spectroscopy, respectively. The optical study has been analyzed using UV-Vis spectroscopy and band gap was calculated as 2.51 eV, using Tauc plot. To analyze and validate the good efficacy of the disc approach, antifungal activity of AgNPs nanoparticles in different concentrations against isolates was achieved in both disc and broth microdilution. The extracellular enzymatic activity of A. fumigatus was found to explore the precise impact of nanoparticles on fungal metabolism. The antifungal efficacy of AgNPs against all fungi was highly successful in disc method. The broth approach underlined the favorable results of the disc method. It provided more precise results in determining the minimum inhibition concentration (MIC), as well as the minimum effective concentration (MEC). A. fumigatus (AM6) enzymatic activity was boosted by AgNPs. Also, ß-galactosidase, ß-glucuronidase, and ß-glucosidase are necessary enzymes whose activity has been boosted. Consequently, M. piperita AgNPs can play a major and intriguing function against resistant Aspergillus species with a significant shift in the enzymatic activity profile of fungi due to this action.

Advances in Membrane-Bound Catechol-O-Methyltransferase Stability Achieved Using a New Ionic Liquid-Based Storage Formulation.

Membrane-bound catechol-O-methyltransferase (MBCOMT), present in the brain and involved in the main pathway of the catechol neurotransmitter deactivation, is linked to several types of human dementia, which are relevant pharmacological targets for new potent and nontoxic inhibitors that have been developed, particularly for Parkinson's disease treatment. However, the inexistence of an MBCOMT 3D-structure presents a blockage in new drugs' design and clinical studies due to its instability. The enzyme has a clear tendency to lose its biological activity in a short period of time. To avoid the enzyme sequestering into a non-native state during the downstream processing, a multi-component buffer plays a major role, with the addition of additives such as cysteine, glycerol, and trehalose showing promising results towards minimizing hMBCOMT damage and enhancing its stability. In addition, ionic liquids, due to their virtually unlimited choices for cation/anion paring, are potential protein stabilizers for the process and storage buffers. Screening experiments were designed to evaluate the effect of distinct cation/anion ILs interaction in hMBCOMT enzymatic activity. The ionic liquids: choline glutamate [Ch][Glu], choline dihydrogen phosphate ([Ch][DHP]), choline chloride ([Ch]Cl), 1- dodecyl-3-methylimidazolium chloride ([C12mim]Cl), and 1-butyl-3-methylimidazolium chloride ([C4mim]Cl) were supplemented to hMBCOMT lysates in a concentration from 5 to 500 mM. A major potential stabilizing effect was obtained using [Ch][DHP] (10 and 50 mM). From the DoE 146% of hMBCOMT activity recovery was obtained with [Ch][DHP] optimal conditions (7.5 mM) at -80 °C during 32.4 h. These results are of crucial importance for further drug development once the enzyme can be stabilized for longer periods of time.

Immune response in sexual inverted and non-inverted Nile tilapia fingerlings supplemented with organic acid and essential oil.

The development and intensification of tilapia farming depends on the manipulation of some physiological functions, such as the sexual inversion of larvae using a synthetic androgen (17α-methyltestosterone). This inversion, however, may represent a potential oxidative stress factor and cause damage to animals in the short, medium, and long term. Dietary supplementation of natural antioxidant compounds is an interesting alternative to combat such damage. To test this hypothesis, an experimental trial was carried out involving sexual inverted and non-inverted Nile tilapia fingerlings, both supplemented and not supplemented with a blend of organic acids and essential oils protected by microencapsulation. Animals were divided into four experimental groups: NI (non-inverted animals), I (sexual inverted animals), NI + M (non-inverted animals supplemented with microcapsules), and I + M (sexual inverted animals supplemented with microcapsules). Blood parameters (WBC - white blood cells; LY - lymphocytes; RBC - red blood cells; HGB - hemoglobin; HCT - hematocrit number; MCH - mean corpuscular hemoglobin; MCV - mean corpuscular volume and MCHC - mean corpuscular hemoglobin concentration), as well as oxidative stress markers (enzymatic activity of superoxide dismutase - SOD and catalase - CAT; and total antioxidant capacity - 2,2-diphenyl-1-picryl-hydrazyl (DPPH)) and gene expression (heat shock protein 70 kDa - HSP70) were evaluated. The HGB (p < 0.001) and HCT (p = 0.005) parameters were reduced beyond the recommended limits for the animals in group I. The MCV varied statistically between the groups (p < 0.001). However, all values were within the recommended range for the species, jointly indicating normocytic anemia in group I fingerlings at the time of collection. The activity of CAT and SOD, as well as DPPH differed statistically between the experimental groups (p < 0.001), with the lowest SOD and CAT activity, as well as the highest DPPH registered in animals supplemented with microcapsules. The expression of HSP70 was lower in I + MI animals (p < 0.001). The synergistic evaluation of the results indicates that animals sexual inverted during the larval stage have a lower total antioxidant capacity in the fingerling stage, which reflects a worsening in hematological and enzymatic parameters related to immunity; and that dietary supplementation with blend of organic acids and essential oils protected by microencapsulation is sufficient to improve the immunological response both in sexual inverted and non-inverted fingerlings.

Unraveling the metabolic shift in anaerobic digestion pathways associated with the alteration of onion skin waste concentration.

Onion skin waste (OSW) is common waste in developing countries, which can cause severe environmental pollution when not properly treated. Value-added products can be chemically extracted from OSW; however, that process is not economically feasible. Alternatively, dry anaerobic digestion (DAD) of OSW is a promising approach for both energy recovery and environment protection. The main hurdles during DAD of OSW can be the hydrolysis and acidification. In batch tests, sludge digestate (SD) rich with methanogens was co-digested with different fractions of OSW for enhancing hydrolysis and raising biogas productivity. The cumulative biogas production (CBP) was 36.6 ± 0.3 mL for sole DAD of SD (100% SD) and increased up to 281.9 ± 14.1 mL for (50% SD: 50% OSW) batch. Self-delignification of OSW took place by SD addition, where the lignin removal reached 75.3 ± 10.5% for (85% SD: 15% OSW) batch. Increasing the fraction of OSW (45% SD: 55% OSW) reduced the delignification by a value of 68.8%, where initial lignin concentration was 9.48 ± 1.6% in dry weight. Lignin breaking down resulted a high fraction of phenolic compounds (345.6 ± 58.8 mg gallic acid equivalent/g dry weight) in the fermentation medium, causing CBP drop (219.0 ± 28.5 mL). The presence of elements (K, Ca, Mg, Fe, Zn, Mn, S and P) in OSW improved the enzymatic activity, facilitated phenolic compounds degradation, shifted the metabolism towards acetate fermentation pathway, and raised biogas productivity. Acidogenesis was less affected by phenolic compounds than methanogenesis, causing higher H2 contents and lower CH4 contents, at batches with high share of OSW.

The DnaJ-like Zinc Finger Protein ORANGE Promotes Proline Biosynthesis in Drought-Stressed Arabidopsis Seedlings.

Orange (OR) is a DnaJ-like zinc finger protein with both nuclear and plastidial localizations. OR, and its orthologs, are highly conserved in flowering plants, sharing a characteristic C-terminal tandem 4× repeats of the CxxCxxxG signature. It was reported to trigger chromoplast biogenesis, promote carotenoid accumulation in plastids of non-pigmented tissues, and repress chlorophyll biosynthesis and chloroplast biogenesis in the nucleus of de-etiolating cotyledons cells. Its ectopic overexpression was found to enhance plant resistance to abiotic stresses. Here, we report that the expression of OR in Arabidopsis thaliana was upregulated by drought treatment, and seedlings of the OR-overexpressing (OE) lines showed improved growth performance and survival rate under drought stress. Compared with the wild-type (WT) and OR-silencing (or) lines, drought-stressed OE seedlings possessed lower contents of reactive oxygen species (such as H2O2 and O2-), higher activities of both superoxide dismutase and catalase, and a higher level of proline content. Our enzymatic assay revealed a relatively higher activity of Δ1-pyrroline-5-carboxylate synthase (P5CS), a rate-limiting enzyme for proline biosynthesis, in drought-stressed OE seedlings, compared with the WT and or lines. We further demonstrated that the P5CS activity could be enhanced by supplementing exogenous OR in our in vitro assays. Taken together, our results indicated a novel contribution of OR to drought tolerance, through its impact on proline biosynthesis.

Modulation of Antioxidant Defense in Farmed Rainbow Trout (Oncorhynchus mykiss) Fed with a Diet Supplemented by the Waste Derived from the Supercritical Fluid Extraction of Basil (Ocimum basilicum).

Phytotherapy is based on the use of plants to prevent or treat human and animal diseases. Recently, the use of essential oils and polyphenol-enriched extracts is also rapidly increasing in the aquaculture sector as a means of greater industrial and environmental sustainability. Previous studies assessed the antibacterial and antiparasitic effects of these bioactive compounds on fish. However, studies on the modulation of oxidative stress biomarkers are still scant to date. Thus, in this study, the modulation of antioxidant defense against oxidative stress exerted by fish diets supplemented with a basil supercritical extract (F1-BEO) was assessed in rainbow trout Oncorhynchus mykiss. The F1-BEO extracted with supercritical fluid extraction was added to the commercial feed flour (0.5, 1, 2, 3% w/w) and mixed with fish oil to obtain a suitable compound for pellet preparation. Fish were fed for 30 days. The levels of stress biomarkers such as superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glyoxalase I, glyoxalase II, lactate dehydrogenase, glutathione and malondialdehyde showed a boost in the antioxidant pathway in fish fed with a 0.5% F1-BEO-supplemented diet. Higher F1-BEO supplementation led to a failure of activity of several enzymes and the depletion of glutathione levels. Malondialdehyde concentration suggests a sufficient oxidative stress defense against lipid peroxidation in all experimental groups, except for a 3% F1-BEO-supplemented diet (liver 168.87 ± 38.79 nmol/mg prot; kidney 146.86 ± 23.28 nmol/mg prot), compared to control (liver 127.76 ± 18.15 nmol/mg prot; kidney 98.68 ± 15.65 nmol/mg prot). Our results suggest supplementing F1-BEO in fish diets up to 0.5% to avoid potential oxidative pressure in farmed trout.