CsSHMT3 gene enhances the growth and development in cucumber seedlings under salt stress.

Salt stress is one of the major factors limiting plant growth and productivity. Many studies have shown that serine hydroxymethyltransferase (SHMT) gene play an important role in growth, development and stress response in plants. However, to date, there have been few studies on whether SHMT3 can enhance salt tolerance in plants. Therefore, the effects of overexpression or silencing of CsSHMT3 gene on cucumber seedling growth under salt stress were investigated in this study. The results showed that overexpression of CsSHMT3 gene in cucumber seedlings resulted in a significant increase in chlorophyll content, photosynthetic rate and proline (Pro) content, and antioxidant enzyme activity under salt stress condition; whereas the content of malondialdehyde (MDA), superoxide anion (H2O2), hydrogen peroxide (O2·-) and relative conductivity were significantly decreased when CsSHMT3 gene was overexpressed. However, the content of chlorophyll and Pro, photosynthetic rate, and antioxidant enzyme activity of the silenced CsSHMT3 gene lines under salt stress were significantly reduced, while MDA, H2O2, O2·- content and relative conductivity showed higher level in the silenced CsSHMT3 gene lines. It was further found that the expression of stress-related genes SOD, CAT, SOS1, SOS2, NHX, and HKT was significantly up-regulated by overexpressing CsSHMT3 gene in cucumber seedlings; while stress-related gene expression showed significant decrease in silenced CsSHMT3 gene seedlings under salt stress. This suggests that overexpression of CsSHMT3 gene increased the salt tolerance of cucumber seedlings, while silencing of CsSHMT3 gene decreased the salt tolerance. In conclusion, CsSHMT3 gene might positively regulate salt stress tolerance in cucumber and be involved in regulating antioxidant activity, osmotic adjustment, and photosynthesis under salt stress. KEY MESSAGE: CsSHMT3 gene may positively regulate the expression of osmotic system, photosynthesis, antioxidant system and stress-related genes in cucumber.

Black Chokeberry (Aronia melanocarpa) Juice Supplementation Improves Oxidative Stress and Aging Markers in Testis of Aged Rats.

Spermatogenesis is a process that continues until the end of an individual's life, although with reduced activity with advancing age. Inflammation, oxidation, and apoptosis are events considered as predictors of pathogenesis and the development of age-related diseases observed in aged testes. The use of natural compounds with antioxidant and anti-inflammatory properties has a beneficial effect on the inflammatory and oxidative status of the aged testis. The aim of this study was to determine the effect of supplementation with antioxidant-rich black chokeberry (Aronia melanocarpa) juice on several markers of oxidative stress and aging in rat testis. In total, 24 male Wistar rats were divided into three experimental groups: young controls aged 2 months, old controls aged 27 months, and 27-month-old rats supplemented with black chokeberry juice at a dose of 10 mL/kg for 3 months. A. melanocarpa juice supplementation led to reduced oxidative stress, manifested by increased immunoexpression of nNOS, eNOS, and MAS1 in the seminiferous tubules and in the Leydig cells. The morphometrically determined tubule structure data showed no significant differences between the three groups. However, the intensity of the immunoreaction for TRK-C and NT3 in Leydig cells was demonstrably higher in the supplemented old animals compared with the old controls. There was a significantly higher number of blood vessels around the seminiferous tubules in the supplemented animals compared to the old controls. These data indicate that supplementation with A. melanocarpa juice slows down aging processes in the testis and preserves the functional activity of Leydig cells.

Biological Activities of Citrus-Derived Extracellular Vesicles on Human Cells: The Role of Preservation.

Extracellular vesicles (EVs) have been identified as important mediators for cell-to-cell communication. Citrus-based EVs in particular offer an excellent platform for nutraceutical delivery systems, as their endemic cargo includes micronutrients (e.g., ascorbic acid), which contribute to their antioxidant capacity. Despite being extensively investigated as to their therapeutic and diagnostic potential, their cargo is inherently unstable and thus directly affected by their storage and preservation. In this study, EVs were isolated from citrus fruit using tangential flow filtration and evaluated for their physicochemical characteristics, antioxidant activity and effects on human cells. To assess how their isolation and preservation methods affect these properties, the EVs were tested immediately after isolation (from fresh and freeze-thawed juices) or following freeze-drying. A measurable biological effect of cryoprotection on citrus-derived EVs was evident, whether during or after isolation. This was more pronounced in the cell-based assays, ranging from -4% to +32% in human skin fibroblast proliferation. Nevertheless, the effects on human cancer cells varied depending on the cell line. Although these results should be considered preliminary observations, subject to further investigation, it is safe to state that any type of preservation is expected to impact the EVs' biological activity.

Nutritional composition, lipid profile and stability, antioxidant activities and sensory evaluation of pasta enriched by linseed flour and linseed oil.

Pasta assortments fortified with high quality foods are a modern nutritional trends. This study, explored the effects of fortification with linseed flour (LF) and linseed oil (LO) on durum wheat pasta characteristics. Wheat flour semolina was replaced with 5%, 10% and 15% of LF or 1%, 2.5% and 5% of LO. Control pasta CP (without LF or LO addition), LF-enriched pasta LFP 5%, LFP 10% and LFP 15% and LO-enriched pasta LOP 1%, LOP 2.5% and LOP 5% was compared for the proteins, fat and phenolic contents and fatty acids (FA) profile. Impact on lipid oxidation and sensory evaluation were also determined. Fortification of pasta with LF improved significantly (p < 0.05) the contents of protein, fat and phenolic compared to CP whereas the enrichment of pasta with LO resulted in a significant increase (p < 0.05) in the content of fat and a significant decrease in protein and phenolic contents. All the formulations decreased the saturated FA percent and increased the polyunsaturated FA percent with enhancement of omega-3 FA content. Antioxidant activity measured by FRAP and DPPH assays was improved after the fortification. For lipid oxidation, the replacement of semolina by LF or LO promoted an increase (p < 0.05) on TBARS values in level-dependent manner. Regarding sensory evaluation, the two types of fortification did not affect the taste; flavor and aroma of cooked pasta, but LOP 5% showed the highest score of the overall acceptability. The results recommended the possibility of producing pasta supplemented with LF or LO (even at a level of 15% and 5% respectively) as a functional food.

PFOA accumulation in the leaves of basil (Ocimum basilicum L.) and its effects on plant growth, oxidative status, and photosynthetic performance.

BACKGROUND: Perfluoroalkyl substances (PFASs) are emerging contaminants of increasing concern due to their presence in the environment, with potential impacts on ecosystems and human health. These substances are considered "forever chemicals" due to their recalcitrance to degradation, and their accumulation in living organisms can lead to varying levels of toxicity based on the compound and species analysed. Furthermore, concerns have been raised about the possible transfer of PFASs to humans through the consumption of edible parts of food plants. In this regard, to evaluate the potential toxic effects and the accumulation of perfluorooctanoic acid (PFOA) in edible plants, a pot experiment in greenhouse using three-week-old basil (Ocimum basilicum L.) plants was performed adding PFOA to growth substrate to reach 0.1, 1, and 10 mg Kg- 1 dw. RESULTS: After three weeks of cultivation, plants grown in PFOA-added substrate accumulated PFOA at different levels, but did not display significant differences from the control group in terms of biomass production, lipid peroxidation levels (TBARS), content of α-tocopherol and activity of ascorbate peroxidase (APX), catalase (CAT) and guaiacol peroxidase (POX) in the leaves. A reduction of total phenolic content (TPC) was instead observed in relation to the increase of PFOA content in the substrate. Furthermore, chlorophyll content and photochemical reflectance index (PRI) did not change in plants exposed to PFAS in comparison to control ones. Chlorophyll fluorescence analysis revealed an initial, rapid photoprotective mechanism triggered by PFOA exposure, with no impact on other parameters (Fv/Fm, ΦPSII and qP). Higher activity of glutathione S-transferase (GST) in plants treated with 1 and 10 mg Kg- 1 PFOA dw (30 and 50% to control, respectively) paralleled the accumulation of PFOA in the leaves of plants exposed to different PFOA concentration in the substrate (51.8 and 413.9 ng g- 1 dw, respectively). CONCLUSION: Despite of the absorption and accumulation of discrete amount of PFOA in the basil plants, the analysed parameters at biometric, physiological and biochemical level in the leaves did not reveal any damage effect, possibly due to the activation of a detoxification pathway likely involving GST.

Allelopathic effect of the methanol extract of the weed species-red sorrel (Rumex acetosella L.) on the growth, phytohormone content and antioxidant activity of the cover crop - white clover (Trifolium repens L.).

Allelopathy is a biological process in which one organism releases biochemicals that affect the growth and development of other organisms. The current investigation sought to determine the allelopathic effect of Rumex acetosella on white clover (Trifolium repens) growth and development by using its shoot extract (lower IC50 value) as a foliar treatment. Here, different concentrations (25, 50, 100, and 200 g/L) of shoot extract from Rumex acetosella were used as treatments. With increasing concentrations of shoot extract, the plant growth parameters, chlorophyll and total protein content of Trifolium repens decreased. On the other hand, ROS, such as O2.- and H2O2, and antioxidant enzymes, including SOD, CAT, and POD, increased with increasing shoot extract concentration. A phytohormonal study indicated that increased treatment concentrations increased ABA and SA levels while JA levels were reduced. For the identification of allelochemicals, liquid‒liquid extraction, thin-layer chromatography, and open-column chromatography were conducted using R. acetosella shoot extracts, followed by a seed bioassay on the separated layer. A lower IC50 value was obtained through GC/MS analysis. gammaSitosterol was identified as the most abundant component. The shoot extract of Rumex acetosella has strong allelochemical properties that may significantly impede the growth and development of Trifolium repens. This approach could help to understand the competitive abilities of this weed species and in further research provide an alternate weed management strategy.

Sustainable remediation of chromium-contaminated soils: boosting radish growth with deashed biochar and strigolactone.

Chromium (Cr) stress significantly hinders crop production by disrupting nutrient uptake, impairing plant growth, and contaminating soil, posing a substantial threat to agricultural sustainability. The use of deashed biochar (DAB) and strigolactone can be an effective solution to mitigate this issue. Deashed biochar enhances crop production by improving soil structure, water retention, and nutrient availability while mitigating the bioavailability of toxic substances. Strigolactone boosts plant growth by stimulating root growth, branching, shoot formation, and overall plant physiology. Nevertheless, the scientific rationale behind their collective use as an amendment to counter Cr stress remains to be substantiated. Therefore, in this study, a blend of DAB and strigolactone was employed as additives in radish cultivation, both in the absence of Cr stress and under the influence of 200Cr stress. Four treatments, i.e., 0, 20µM Strigolactone, DAB, and 20µM Strigolactone + DAB, were applied in four replications following a completely randomized design. Results demonstrate that 20µM Strigolactone + DAB produced significant improvement in radish shoot length (27.29%), root length (45.60%), plant fresh weight (33.25%), and plant dry weight (78.91%), compared to the control under Cr stress. Significant enrichment in radish chlorophyll a (20.41%), chlorophyll b (58.53%), and total chlorophyll (31.54%) over the control under Cr stress, prove the efficacy of 20µM Strigolactone + DAB treatment. In conclusion, 20µM Strigolactone + DAB is the recommended amendment for mitigating Cr stress in radish. Farmers should consider using Strigolactone + DAB amendments to combat Cr stress and enhance radish growth, contributing to a more resilient agricultural ecosystem.

Enhancing french basil growth through synergistic Foliar treatment with copper nanoparticles and Spirulina sp.

BACKGROUND: This study investigates a novel idea about the foliar application of nanoparticles as nanofertilizer combined with a natural stimulant, blue-green algae Spirulina platensis L. extract, as a bio-fertilizer to achieve safety from using nanoparticles for enhancement of the growth and production of the plant. Thus, this experiment aimed to chemically synthesize copper nanoparticles via copper sulfate in addition to evaluate the impact of CuNPs at 500, 1000, and 1500 mg/L and the combination of CuNPs with or without microalgae extract at 0.5, 1, and 1.5 g/L on the morphological parameters, photosynthetic pigments accumulation, essential oil production, and antioxidant activity of French basil. RESULTS: The results revealed that foliar application of CuNPs and its interaction with spirulina extract significantly increased growth and yield compared with control, the treatments of 1000 and 1500 mg/L had less impact than 500 mg/L CuNPs. Plants treated with 500 mg/L CuNPs and 1.5 g/L spirulina extract showed the best growth and oil production, as well as the highest accumulation of chlorophylls and carotenoids. The application of CuNPs nanofertilizer caused a significant increase in the antioxidant activity of the French basil plant, but the combination of CuNPs with spirulina extract caused a decrease in antioxidant activity. CONCULOSION: Therefore, foliar application of natural bio-fertilizer with CuNPsis necessary for obtaining the best growth and highest oil production from the French basil plant with the least damage to the plant and the environment.

Effect of preharvest biofilm application regimes on cracking and fruit quality traits in '0900 Ziraat' sweet cherry cultivar.

BACKGROUND: Fruit cracking impacts the quality of sweet cherry, significantly affecting its marketability due to increased susceptibility to injury, aesthetic flaws, and susceptibility to pathogens. The effect of 1% biofilm (Parka™) application regimes on fruit cracking and other quality parameters in the '0900 Ziraat' cherry cultivar was investigated in this study. Fruit sprayed with water were served as control (U1). Fruit treated only once with biofilm three, two and one week before the commercial harvest were considered as U2, U3 and U4, respectively. Fruit treated with biofilm three, two, and one week before harvest were considered as U5; three and two week before harvest as U6; two and one week before harvest as U7; and fruit treated three and one week before harvest as U8. RESULTS: In both measurement periods, the lower cracking index was obtained in biofilm-treated sweet cherry fruit. However, the firmness of biofilm-treated fruit was higher than that of the control fruit. The lowest respiration rate was observed in U7, while the highest weight was recorded in U4 and U5 than the control. The biofilm application decreased fruit coloration. The biofilm application also increased the soluble solids content of the fruit. The U2, U3 and U4 applications at harvest showed higher titratable acidity than the control. In both measurement periods, the vitamin C content of the U2, U5, U6, U7 and U8 applications was found to be higher than that of the control. The total monomeric anthocyanin of the U3 and U8 applications was higher than that of the control. Furthermore, the antioxidant activity of the U2, U3 and U5 in the DPPH, and the U7 and U8 in FRAP were measured higher thanthat of the control. CONCLUSIONS: The application of biofilms has the potential to mitigate fruit cracking, prolong postharvest life of sweet cherries, and enhance fruit firmness.

Comparative morpho-physiological and biochemical responses of Capsicum annuum L. plants to multi-walled carbon nanotubes, fullerene C60 and graphene nanoplatelets exposure under water deficit stress.

Water deficit stress is one of the most significant environmental abiotic factors influencing plant growth and metabolism globally. Recently, encouraging outcomes for the use of nanomaterials in agriculture have been shown to reduce the adverse effects of drought stress on plants. The present study aimed to investigate the impact of various carbon nanomaterials (CNMs) on the physiological, morphological, and biochemical characteristics of bell pepper plants subjected to water deficit stress conditions. The study was carried out as a factorial experiment using a completely randomized design (CRD) in three replications with a combination of three factors. The first factor considered was irrigation intensity with three levels [(50%, 75%, and 100% (control) of the field capacity (FC)] moisture. The second factor was the use of carbon nanomaterials [(fullerene C60, multi-walled carbon nanotubes (MWNTs) and graphene nanoplatelets (GNPs)] at various concentrations [(control (0), 100, 200, and 1000 mg/L)]. The study confirmed the foliar uptake of CNMs using the Scanning Electron Microscopy (SEM) technique. The effects of the CNMs were observed in a dose-dependent manner, with both stimulatory and toxicity effects being observed. The results revealed that exposure to MWNTs (1000 mg/L) under well-watered irrigation, and GNPs treatment (1000 mg/L) under severe drought stress (50% FC) significantly (P < 0.01) improved fruit production and fruit dry weight by 76.2 and 73.2% as compared to the control, respectively. Also, a significant decrease (65.9%) in leaf relative water content was obtained in plants subjected to soil moisture of 50% FC over the control. Treatment with GNPs at 1000 mg/L under 50% FC increased electrolyte leakage index (83.6%) compared to control. Foliar applied MWNTs enhanced the leaf gas exchange, photosynthesis rate, and chlorophyll a and b concentrations, though decreased the oxidative shock in leaves which was demonstrated by the diminished electrolyte leakage index and upgrade in relative water content and antioxidant capacity compared to the control. Plants exposed to fullerene C60 at 100 and 1000 mg/L under soil moisture of 100 and 75% FC significantly increased total flavonoids and phenols content by 63.1 and 90.9%, respectively, as compared to the control. A significant increase (184.3%) in antioxidant activity (FRAP) was observed in plants exposed to 200 mg/L MWCNTs under irrigation of 75% FC relative to the control. The outcomes proposed that CNMs could differentially improve the plant and fruit characteristics of bell pepper under dry conditions, however, the levels of changes varied among CNMs concentrations. Therefore, both stimulatory and toxicity effects of employed CNMs were observed in a dose-dependent manner. The study concludes that the use of appropriate (type/dose) CNMs through foliar application is a practical tool for controlling the water shortage stress in bell pepper. These findings will provide the basis for more research on CNMs-plant interactions, and with help to ensure their safe and sustainable use within the agricultural chains.

Identification of a novel gene PpBCG1 that has a positive role in desiccation tolerance in the moss Physcomitrium patens.

Water-to-land transition is a hallmark of terrestrialization for land plants and requires molecular adaptation to resist water deficiency. Lineages- or species-specific genes are widespread across eukaryotes, and yet the majority of those are functionally unknown and not annotated. Recent studies have revealed that some of such genes could play a role in adapting to environmental stress responses. Here, we identified a novel gene PpBCG1 (Bryophyte Co-retained Gene 1) in the moss Physcomitrium patens that was responsive to dehydration and rehydration. Under de- and rehydration treatments, PpBCG1 was significantly co-expressed with the dehydrin-encoding gene PpDHNA. Microarray data revealed that PpBCG1 was highly expressed in tissues of spores, female organ archegonia, and mature sporophytes. In addition, the Ppbcg1 mutant showed reduced ability of dehydration tolerance, whose plants were accompanied by a relatively low level of chlorophyll content during recovery. Comprehensive transcriptomics uncovered a detailed set of regulatory processes that were affected by the PpBCG1 disruption. Moreover, experimental evidence showed that PpBCG1 might function in the antioxidant activity, abscisic acid (ABA) pathway, and intracellular calcium (Ca2+) homeostasis to resist desiccation. Together, our study provides insights into the roles of one bryophyte co-retained gene in the desiccation tolerance.

Acanthopanax senticosus cultures fermented by Lactobacillus rhamnosus enhanced immune response through improvement of antioxidant activity and inflammation in crucian carp (Carassius auratus).

Lactic acid bacteria (LAB) have been recognized as safe microorganism that improve micro-flora disturbances and enhance immune response. A well-know traditional herbal medicine, Acanthopanax senticosus (As) was extensively utilized in aquaculture to improve growth performance and disease resistance. Particularly, the septicemia, skin wound and gastroenteritis caused by Aeromonas hydrophila threaten the health of aquatic animals and human. However, the effects of probiotic fermented with A. senticosus product on the immune regulation and pathogen prevention in fish remain unclear. Here, the aim of the present study was to elucidate whether the A. senticosus fermentation by Lactobacillus rhamnosus improve immune barrier function. The crucian carp were fed with basal diet supplemented with L. rhamnosus fermented A. senticosus cultures at 2 %, 4 %, 6 % and 8 % bacterial inoculum for 8 weeks. After trials, the weight gain rate (WGR), specific growth rate (SGR) were significantly increased, especially in LGG-6 group. The results confirmed that the level of the CAT, GSH-PX, SOD, lysozyme, and MDA was enhanced in fish received with probiotic fermented product. Moreover, the L. rhamnosus fermented A. senticosus cultures could trigger innate and adaptive immunity, including the up-regulation of the C3, C4, and IgM concentration. The results of qRT-PCR revealed that stronger mRNA transcription of IL-1ß, IL-10, IFN-γ, TNF-α, and MyD88 genes in the liver, spleen, kidney, intestine and gills tissues of fish treated with probiotic fermented with A. senticosus product. After infected with A. hydrophila, the survival rate of the LGG-2 (40 %), LGG-4 (50 %), LGG-6 (60 %), LGG-8 (50 %) groups was higher than the control group. Meanwhile, the pathological damage of the liver, spleen, head-kidney, and intestine tissues of probiotic fermentation-fed fish could be alleviated after pathogen infection. Therefore, the present work indicated that L. rhamnosus fermented A. senticosus could be regard as a potential intestine-target therapy strategy to protecting fish from pathogenic bacteria infection.

Synthesis and characterization of marine seagrass (Cymodocea serrulata) mediated titanium dioxide nanoparticles for antibacterial, antibiofilm and antioxidant properties.

Cymodocea serrulata mediated titanium dioxide nanoparticles (TiO2 NPs) were successfully synthesized. The XRD pattern and FTIR spectra demonstrated the crystalline structure of TiO2 NPs and the presence of phenols, flavonoids and alkaloids in the extract. Further SEM revealed that TiO2 NPs has uniform structure and spherical in shape with their size ranged from 58 to 117 nm. Antibacterial activity of TiO2 NPs against methicillin-resistant Staphylococcus aureus (MRSA) and Vibrio cholerae (V. cholerae), provided the zone of inhibition of 33.9 ± 1.7 and 36.3 ± 1.9 mm, respectively at 100 µg/mL concentration. MIC of TiO2 NPs against MRSA and V. cholerae showed 84% and 87% inhibition at 180 µg/mL and 160 µg/mL respectively. Subsequently, the sub-MIC of V. cholerae demonstrated minimal or no impact on bacterial growth at concentration of 42.5 µg/mL concentration. In addition, TiO2 NPs exhibited their ability to inhibit the biofilm forming V. cholerae which caused distinct morphological and intercellular damages analysed using CLSM and TEM. The antioxidant properties of TiO2 NPs were demonstrated through TAA and DPPH assays and exposed its scavenging activity with IC50 value of 36.42 and 68.85 µg/mL which denotes its valuable antioxidant properties with potential health benefits. Importantly, the brine shrimp based lethality experiment yielded a low cytotoxic effect with 13% mortality at 100 µg/mL. In conclusion, the multifaceted attributes of C. serrulata mediated TiO2 NPs encompassed the antibacterial, antioxidant and anti-biofilm inhibition effects with low cytotoxicity in nature were highlighted in this study and proved the bioderived TiO2 NPs could be used as a promising agent for biomedical applications.

Involvement of the Keap1-Nrf2-ARE pathway in the antioxidant activity of sinomenine.

Sinomenine is a pure alkaloid isolated from Sinomenium acutum. This study is aimed to investigate the critical role of the nuclear factor erythroid 2-related factor 2 (Nrf2)-kelch-like ECH-associated protein-1(Keap1)-antioxidant response element (ARE) antioxidative signaling pathway in protecting sinomenine against H2O2-induced oxidative injury. Cytotoxicity and antioxidant experiments to initially determine the protective effects of sinomenine show that sinomenine has no effect on the decreased cell viability and presents similar potency in scavenging all three free radicals. The binding affinity between sinomenine and Keap1 was determined via fluorescence polarization assay, with IC50 of 13.52 µM. Quantum chemical calculation and theoretical simulation illustrated that sinomenine located into the Nrf2-binding site of Keap1 via hydrophobic and hydrogen interactions, showing high stability and binding affinity. On the basis of the stable binding of sinomenine with Keap1, sinomenine efficiently induced nuclear translocation of Nrf2, and increased in ARE activity in a concentration-dependent manner. Quantitative polymerase chain reaction provided further evidences that sinomenine-induced protection upregulated ARE-dependent genes, such as NAD(P)H quinone oxidoreductase 1, hemeoxygenase-1, and glutamate-cysteine ligase modifier subunit. Western blot confirmed that sinomenine increased the expressions of these antioxidative enzymes. Taken together, in vitro and in silico evaluations demonstrate that sinomenine inhibits the binding of Keap1 to Nrf2, promotes the nuclear accumulation of Nrf2 and thus leads to the upregulated expressions of Nrf2-dependent antioxidative genes. Our findings also highlight the use of sinomenine for pharmacological or therapeutic regulation of the Nrf2-Keap1-ARE system, which is a novel strategy to prevent the progression of oxidative injury.

Neuroprotective and Anti-inflammatory Effects of Rubiscolin-6 Analogs with Proline Surrogates in Position 2.

Naturally occurring peptides, such as rubiscolins derived from spinach leaves, have been shown to possess some interesting activities. They exerted central effects, such as antinociception, memory consolidation and anxiolytic-like activity. The fact that rubiscolins are potent even when given orally makes them very promising drug candidates. The present work tested whether rubiscolin-6 (R-6, Tyr-Pro-Leu-Asp-Leu-Phe) analogs have neuroprotective and anti-inflammatory effects. These hypotheses were tested in the 6-hydroxydopamine (6-OHDA) injury model of human neuroblastoma SH-SY5Y and lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The determination of reactive oxygen species (ROS), mitochondrial membrane potential (MMP), Caspase-3 activity, lipid peroxidation and nitric oxide (NO) production allowed us to determine the effects of peptides on hallmarks related to Parkinson's Disease (PD) and inflammation. Additionally, we investigated the impact of R-6 analogs on serine-threonine kinase (also known as protein kinase B, AKT) and mammalian target of rapamycin (mTOR) activation. The treatment with analogs 3 (Tyr-Inp-Leu-Asp-Leu-Phe-OH), 5 (Dmt-Inp-Leu-Asp-Leu-Phe-OH) and 7 (Tyr-Inp-Leu-Asp-Leu-Phe-NH2) most effectively prevented neuronal death via attenuation of ROS, mitochondrial dysfunction and Caspase-3 activity. Peptides 5 and 7 significantly increased the protein expression of the phosphorylated-AKT (p-AKT) and phosphorylated-mTOR (p-mTOR). Additionally, selected analogs could also ameliorate LPS-mediated inflammation in macrophages via inhibition of intracellular generation of ROS and NO production. Our findings suggest that R-6 analogs exert protective effects, possibly related to an anti-oxidation mechanism in in vitro model of PD. The data shows that the most potent peptides can inhibit 6-OHDA injury by activating the PI3-K/AKT/mTOR pathway, thus playing a neuroprotective role and may provide a rational and robust approach in the design of new therapeutics or even functional foods.

Antioxidant compounds produced by endolichenic fungus Penicillium sp.-strain 1322P isolated from Pyxine subcinerea.

Endolichenic fungi are expecting for new bioresources of pharmacological compounds. However, the number of investigations targeting antioxidant compounds produced by endolichenic fungi remains limited. To discover new antioxidant compounds, we analyzed the antioxidant activity of the methanol extracts derived from isolated lichen mycobionts or endolichenic fungi induced from Pyxine subcinerea. We performed this analysis using the oxygen radical absorbance capacity (ORAC) method. As a result, we isolated from an endolichenic fungus identified as Penicillium sp.-stain 1322P in Pyxine subcinerea. This fungus produced a red pigment, and its chemical structure was determined to be sclerotioramine based on the analytical data obtained from NMR, LC-MS/MS, and HPLC-PDA. Sclerotioramine exhibited high antioxidant activity, and the ORAC values (mean ± SD) of sclerotioramine and sclerotiorin were 11.4 ± 0.36 and 4.86 ± 0.70 mmol TE per gram of the respective pure compound. Thus, the antioxidant activity of sclerotioramine was greater than twice that of sclerotiorin. This work represents the first report that the antioxidant activity of sclerotioramine is higher than that of the sclerotiorin.

Assessment of the in-vitro probiotic efficacy and safety of Pediococcus pentosaceus L1 and Streptococcus thermophilus L3 isolated from Laban, a popular fermented milk product.

Probiotics are beneficial microorganisms, mostly lactic acid bacteria (LAB), that offer health benefits to the host when consumed in adequate amounts. This study assessed the probiotic efficacy and safety of LAB strains isolated from Laban, a traditional fermented milk product. Seven primarily selected Gram-positive, catalase-negative, non-spore-forming isolates were examined for their antimicrobial activity against the bacterial pathogens Bacillus cereus, Salmonella typhi, Staphylococcus aureus, and Vibrio cholera, and the fungal pathogen Candida albicans. Two isolates, identified as Pediococcus pentosaceus L1 and Streptococcus thermophilus L3, which showed antimicrobial activity against all pathogens, were further evaluated for their probiotic competence. The selected isolates demonstrated strong resistance to low pH, bile salts, and phenol, indicating their potential for gastric endurance. They also exhibited high cell surface hydrophobicity to various hydrocarbons, autoaggregation, and coaggregation properties, demonstrating strong adhesion abilities. In addition, both isolates showed strong antioxidant activity and were non-hemolytic. Although the isolates had some resistance to certain antibiotics, they were generally susceptible to commonly used antibiotics. The two LAB strains also exhibited promising technological properties, such as milk coagulation and exopolysaccharide production, indicating their potential to enhance the quality of dairy products. The results suggest that the LAB strains isolated from Laban have strong potential as probiotics, and due to their food origin, they are highly likely to exhibit maximal efficacy in food and pharmaceutical products for human consumption.

The colorful fungi of the Chilean forests: Production, chemical characterization and possible applications of their pigments.

In Chile, as in the rest of the world, only a small fraction of the fungal diversity inhabiting the wide variety of its ecosystems is known. This diversity must hide an inestimable richness of species with interesting biotechnological potential, including fungal pigment producers. Recently, interest in filamentous fungi has increased significantly due to their importance as alternative sources of pigments and colorants that are environmentally and human health friendly. As a result, fungal pigments are gaining importance in various industrial applications, such as food, textiles, pharmaceuticals, cosmetics, etc. The increasing consumer demand for "green label" natural colorants requires the exploration of different ecosystems in search of new fungal species that are efficient producers of different pigment with a wide range of colors and ideally without the co-production of mycotoxins. However, advances are also needed in pigment production processes through fermentation, scale-up from laboratory to industrial scale, and final product formulation and marketing. In this respect, the journey is still full of challenges for scientists and entrepreneurs. This chapter describes studies on pigment-producing fungi collected in the forests of central-southern Chile. Aspects such as the exploration of potential candidates as sources of extracellular pigments, the optimization of pigment production by submerged fermentation, methods of pigment extraction and purification for subsequent chemical characterization, and formulation (by microencapsulation) for potential cosmetic applications are highlighted. This potential use is due to the outstanding bioactivity of most fungal pigments, making them interesting functional ingredients for many applications. Finally, the use of fungal pigments for textile and spalting applications is discussed.

Mapping a sustainable approach: biosynthesis of lactobacilli-silver nanocomposites using whey-based medium for antimicrobial and bioactivity applications.

This study explores a sustainable approach for synthesizing silver nanocomposites (AgNCs) with enhanced antimicrobial and bioactivity using safe Lactobacillus strains and a whey-based medium (WBM). WBM effectively supported the growth of Lactobacillus delbrueckii and Lactobacillus acidophilus, triggering a stress response that led to AgNCs formation. The synthesized AgNCs were characterized using advanced spectroscopic and imaging techniques such as UV‒visible, Fourier transform infrared (FT-IR) spectroscopy, transmission electron (TEM), and scanning electron microscopy with energy dispersive X-ray analysis (SEM-Edx). Lb acidophilus-synthesized AgNCs in WBM (had DLS size average 817.2-974.3 ± PDI = 0.441 nm with an average of metal core size 13.32 ± 3.55 nm) exhibited significant antimicrobial activity against a broad spectrum of pathogens, including bacteria such as Escherichia coli (16.47 ± 2.19 nm), Bacillus cereus (15.31 ± 0.43 nm), Clostridium perfringens (25.95 ± 0.03 mm), Enterococcus faecalis (32.34 ± 0.07 mm), Listeria monocytogenes (23.33 ± 0.05 mm), methicillin-resistant Staphylococcus aureus (MRSA) (13.20 ± 1.76 mm), and filamentous fungi such as Aspergillus brasiliensis (33.46 ± 0.01 mm). In addition, Lb acidophilus-synthesized AgNCs in WBM exhibit remarkable free radical scavenging abilities, suggesting their potential as bioavailable antioxidants. These findings highlight the dual functionality of these biogenic AgNCs, making them promising candidates for applications in both medicine and nutrition.

Fate of melatonin orally administered in preterm newborns: Antioxidant performance and basis for neuroprotection.

Preterm infants cannot counteract excessive reactive oxygen species (ROS) production due to preterm birth, leading to an excess of lipid peroxidation with malondialdehyde (MDA) production, capable of contributing to brain damage. Melatonin (ME), an endogenous brain hormone, and its metabolites, act as a free radical scavenger against ROS. Unfortunately, preterms have an impaired antioxidant system, resulting in the inability to produce and release ME. This prospective, multicenter, parallel groups, randomized, double-blind, placebo-controlled trial aimed to assess: (i) the endogenous production of ME in very preterm infants (gestational age ≤ 29 + 6 WE, 28 infants in the ME and 26 in the placebo group); (ii) the exogenous hormone availability and its metabolization to the main metabolite, 6-OH-ME after 15 days of ME oral treatment; (iii) difference of MDA plasma concentration, as peroxidation marker, after treatment. Blood was collected before the first administration (T1) and after 15 days of administration (T2). ME and 6-OH-ME were detected by liquid chromatography tandem mass spectrometry, MDA was measured by liquid chromatograph with fluorescence detection. ME and 6-OH-ME were not detectable in the placebo group at any study time-point. ME was absent in the active group at T1. In contrast, after oral administration, ME and 6-OH-ME resulted highly detectable and the difference between concentrations T2 versus T1 was statistically significant, as well as the difference between treated and placebo groups at T2. MDA levels seemed stable during the 15 days of treatment in both groups. Nevertheless, a trend in the percentage of neonates with reduced MDA concentration at T2/T1 was 48.1% in the ME group versus 38.5% in the placebo group. We demonstrated that very preterm infants are not able to produce endogenous detectable plasma levels of ME during their first days of life. Still, following ME oral administration, appreciable amounts of ME and 6-OH-ME were available. The trend of MDA reduction in the active group requires further clinical trials to fix the dosage, the length of ME therapy and to identify more appropriate indexes to demonstrate, at biological and clinical levels, the antioxidant activity and consequent neuroprotectant potential of ME in very preterm newborns.