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Introduction: Triple negative breast cancer (TNBC), a highly aggressive subtype accounting for 15–20% of all breast cancer cases, faces limited treatment options often accompanied by severe side effects. In recent years, natural extracellular nanovesicles derived from plants have emerged as promising candidates for cancer therapy, given their safety profile marked by non-immunogenicity and absence of inflammatory responses. Nevertheless, the potential anti-cancer effects of Citrus limonL.-derived extracellular nanovesicles (CLENs) for breast cancer treatment is still unexplored.Methods: In this study, we investigated the anti-cancer effects of CLENs on two TNBC cell lines (4T1 and HCC-1806 cells) under growth conditions in 2D and 3D culture environments. The cellular uptake efficiency of CLENs and their internalization mechanism were evaluated in both cells using confocal microscopy. Thereafter, we assessed the effect of different concentrations of CLENs on cell viability over time using a dual approach of Calcein-AM PI live-dead assay and CellTiter-Glo bioluminescence assay. We also examined the influence of CLENs on the migratory and evasion abilities of TNBC cells through wound healing and 3D Matrigel drop evasion assays. Furthermore, Western blot analysis was employed to investigate the effects of CLENs on the phosphorylation levels of phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and extracellular signal- regulated kinase (ERK) expression.Results: We found that CLENs were internalized by the cells via endocytosis, leading to decreased cell viability, in a dose- and time-dependent manner. Additionally, the migration and evasion abilities of TNBC cells were significantly inhibited under exposed to 40 and 80 μg/mL CLENs. Furthermore, down-regulated expression levels of phosphorylated phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and extracellular signal-regulated kinase (ERK), suggesting that the inhibition of cancer cell proliferation, migration, and evasion is driven by the inhibition of the PI3K/AKT and MAPK/ERK signaling pathways.Discussion: Overall, our results demonstrate the anti-tumor efficiency of CLENs against TNBC cells, highlighting their potential as promising natural anti-cancer agents for clinical applications in cancer treatment.

Plant-derived nanovesicles (PDNVs) have recently emerged as natural delivery systems of biofunctional compounds toward mammalian cells. Considering their already described composition, anti-inflammatory properties, stability, and low toxicity, PDNVs offer a promising path for developing new preventive strategies for several inflammatory diseases, among which the inflammatory bowel disease (IBD). In this study, we explore the protective effects of industrially produced lemon vesicles (iLNVs) in a rat model of IBD. Characterization of iLNVs reveals the presence of small particles less than 200 nm in size and a profile of bioactive compounds enriched in flavonoids and organic acids with known beneficial properties. In vitro studies on human macrophages confirm the safety and anti-inflammatory effects of iLNVs, as evidenced by the reduced expression of pro-inflammatory cytokines and increased levels of anti-inflammatory markers. As evidenced by in vivo experiments, pre-treatment with iLNVs significantly alleviates symptoms and histological features in 2,4 dinitrobenzensulfuric acid (DNBS)-induced colitis in rats. Molecular pathway analysis reveals modulation of NF-κB and Nrf2, indicating anti-inflammatory and antioxidant effects. Finally, iLNVs affects gut microbiota composition, improving the consistent colitis-related alterations. Overall, we demonstrated the protective role of industrially produced lemon nanovesicles against colitis and emphasized their potential in managing IBD through multifaceted mechanisms.

Chronic inflammation is associated with the occurrence of several diseases. However, the side effects of anti‐inflammatory drugs prompt the identification of new therapeutic strategies. Plant‐derived extracellular vesicles (PDEVs) are gaining increasing interest in the scientific community for their biological properties. We isolated PDEVs from the juice of Citrus limon L. (LEVs) and characterized their flavonoid, limonoid and lipid contents through reversed‐phase high‐performance liquid chromatography coupled to electrospray ionization quadrupole time‐of‐flight mass spectrometry (RP‐HPLC–ESI‐Q‐TOF‐MS). To investigate whether LEVs have a protective role on the inflammatory process, murine and primary human macrophages were pre‐treated with LEVs for 24 h and then were stimulated with lipopolysaccharide (LPS). We found that pre‐treatment with LEVs decreased gene and protein expression of pro‐inflammatory cytokines, such as IL‐6, IL1‐β and TNF‐α, and reduced the nuclear translocation and phosphorylation of NF‐κB in LPS‐stimulated murine macrophages. The inhibition of NF‐κB activation was associated with the reduction in ERK1‐2 phosphorylation. Furthermore, the ability of LEVs to decrease pro‐inflammatory cytokines and increase anti‐inflammatory molecules was confirmed ex vivo in human primary T lymphocytes. In conclusion, we demonstrated that LEVs exert anti‐inflammatory effects both in vitro and ex vivo by inhibiting the ERK1‐2/NF‐κB signalling pathway. [ABSTRACT FROM AUTHOR]

Introduction: Triple negative breast cancer (TNBC), a highly aggressive subtype accounting for 15-20% of all breast cancer cases, faces limited treatment options often accompanied by severe side effects. In recent years, natural extracellular nanovesicles derived from plants have emerged as promising candidates for cancer therapy, given their safety profile marked by non-immunogenicity and absence of inflammatory responses. Nevertheless, the potential anti-cancer effects of Citrus limon L .-derived extracellular nanovesicles (CLENs) for breast cancer treatment is still unexplored. Methods: In this study, we investigated the anti-cancer effects of CLENs on two TNBC cell lines (4T1 and HCC-1806 cells) under growth conditions in 2D and 3D culture environments. The cellular uptake efficiency of CLENs and their internalization mechanism were evaluated in both cells using confocal microscopy. Thereafter, we assessed the effect of different concentrations of CLENs on cell viability over time using a dual approach of Calcein-AM PI live-dead assay and CellTiter-Glo bioluminescence assay. We also examined the influence of CLENs on the migratory and evasion abilities of TNBC cells through wound healing and 3D Matrigel drop evasion assays. Furthermore, Western blot analysis was employed to investigate the effects of CLENs on the phosphorylation levels of phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and extracellular signal- regulated kinase (ERK) expression. Results: We found that CLENs were internalized by the cells via endocytosis, leading to decreased cell viability, in a dose- and time-dependent manner. Additionally, the migration and evasion abilities of TNBC cells were significantly inhibited under exposed to 40 and 80 μg/mL CLENs. Furthermore, down-regulated expression levels of phosphorylated phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and extracellular signal-regulated kinase (ERK), suggesting that the inhibition of cancer cell proliferation, migration, and evasion is driven by the inhibition of the PI3K/AKT and MAPK/ERK signaling pathways. Discussion: Overall, our results demonstrate the anti-tumor efficiency of CLENs against TNBC cells, highlighting their potential as promising natural anti-cancer agents for clinical applications in cancer treatment., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Cui, Perini, Augello, Palmieri, De Spirito and Papi.)

Plant-derived nanovesicles (PDNVs) have recently emerged as natural delivery systems of biofunctional compounds toward mammalian cells. Considering their already described composition, anti-inflammatory properties, stability, and low toxicity, PDNVs offer a promising path for developing new preventive strategies for several inflammatory diseases, among which the inflammatory bowel disease (IBD). In this study, we explore the protective effects of industrially produced lemon vesicles (iLNVs) in a rat model of IBD. Characterization of iLNVs reveals the presence of small particles less than 200 nm in size and a profile of bioactive compounds enriched in flavonoids and organic acids with known beneficial properties. In vitro studies on human macrophages confirm the safety and anti-inflammatory effects of iLNVs, as evidenced by the reduced expression of pro-inflammatory cytokines and increased levels of anti-inflammatory markers. As evidenced by in vivo experiments, pre-treatment with iLNVs significantly alleviates symptoms and histological features in 2,4 dinitrobenzensulfuric acid (DNBS)-induced colitis in rats. Molecular pathway analysis reveals modulation of NF-κB and Nrf2, indicating anti-inflammatory and antioxidant effects. Finally, iLNVs affects gut microbiota composition, improving the consistent colitis-related alterations. Overall, we demonstrated the protective role of industrially produced lemon nanovesicles against colitis and emphasized their potential in managing IBD through multifaceted mechanisms. [Display omitted] • Lemon nanovesicles (iLNVs) play a protective role against DNBS-induced colitis in rat. • iLNVs decrease colitis-associated inflammatory damages. • iLNVs restore the rat intestinal permeability acting on tight junctions. • iLNVs shape the gut microbiota. [ABSTRACT FROM AUTHOR]

The present study reports a benign approach to prepare citrate capped iron particles using juice of a lemon fruit. Resultant particles were subjected to detailed adsorption studies for removal of arsenite aqueous solution of defined concentration. The planer reflection of XRD analysis suggested that iron particles were crystalline in nature. FT-IR study suggested that functional groups like -NH2, -NO2, -CO, and --CS were involved in arsenite ion sorption. The composite removal efficiency for As (III) was found to be 88.58%, at pH 4.5, with contact time of 30 min and temperature at 30°C. Data of adsorption were analyzed using various parameter like isotherm models, thermodynamic and kinetic models. Chemisorptions resulted in to formation of monolayer arsenite ions over the surface of prepared iron particles with maximum adsorption capacity; 55.53 mg/gm. Adsorbent hold good promises to remove arsenic from contaminated water. [ABSTRACT FROM AUTHOR]

The experiment was carried out in the main orchard of the Horticulture Department / Al-Hindiya Station for the production of certified citrus seedlings / Karbala Governorate / Iraqi Ministry of Agriculture, during the 2018 season. The experiment included 9 treatments and three replicates to study the interaction between two factors, The first factor was Optimus plus nutrient solution in three concentrations (0, 1, 2 ml. L-1) which contains (5% nitrogen, 30% amino acid and 3% organic nitrogen) and the second factor was Fe-chelate with three concentrations (0, 500, 1000 mg.L-1). Local lemon grafted on sour were sprayed at a rate of three applications (before sour, month after the last spray). The results showed that spraying with nano-fertilizer Optimus plus at concentration of 2 ml.L-1 and Fe-chelate a concentration of 1000 mg.L-1 resulted in the highest increase in all studied traits including the leaf area, the leaf content of total chlorophyll, and carbohydrates, the number of fruits, fruit weight and total yield, with significant differences compared to the control treatment. [ABSTRACT FROM AUTHOR]

Background & Objective: One of the plants used in traditional medicine is lemon which has analgesic effect. However, little research has been performed on the analgesic effect of lemon and mechanisms of action with an emphasis on neurotransmitters systems. Therefore, the present study set to investigate the peripheral effects of lemon essential oil on somatic pain using formalin test with an emphasis on histaminergic system in male Wistar rats. Materiala & Methods: Sixty male rats weighing approximately 200-250g and aged 14-16 wk were divided into 10 groups: sham (Salin + Formalin 1% intraplantar), three treatment groups with lemon essential oil (EO) (12.5, 25 and 50 mg/kg), three treatment groups with Chlorpheniramine (5, 10 and 20 mg/kg), 1 treatment group with Histamine (10 mg/kg), 1 pretreatment group with Chlorpheniramine (20 mg/kg) + EO (50mg/kg), and 1 pretreatment group with Histamine (10 mg/kg) + EO (50 mg/kg). Formalin test was used to assess somatic pain. Data analysis was performed using one-way ANOVA. Results: Intraperitoneal injection of lemon essential oil reduced the pain response induced by formalin in both phases (P

This study aimed to evaluate the in vitro antifungal activity (AA) of the essential oil (EO) of lemon ( Citrus limon L.) against three pathogenic fungi attacking grapevine wood. The composition of the EO was also studied. Ten volatile components were identified by gas chromatography–mass spectrometry. The results showed that the EO consists of volatile components where monoterpene hydrocarbons are the most abundant ones. Four major components were identified, which represent 99.9% of the total EO (limonene, neral, β-pinene, and γ-terpinene). The AA of the EO was evaluated against three pathogenic fungi attacking grapevine wood ( Eutypa sp., Botryosphaeria dothidea , and Fomitiporia mediterranea ). The results showed that the EO exerts AA against all tested fungi and significantly inhibits their growth. Eutypa sp. is the most sensitive fungus. These results show, for the first time, a new use for the EO of lemon ( C. limon L.) to control fungal diseases of grapevine wood. [ABSTRACT FROM AUTHOR]

Background & Objective: One of the plants used in traditional medicine is lemon which has analgesic effect. However, little research has been performed on the analgesic effect of lemon and mechanisms of action with an emphasis on neurotransmitters systems. Therefore, the present study set to investigate the peripheral effects of lemon essential oil on somatic pain using formalin test with an emphasis on histaminergic system in male Wistar rats. Materiala & Methods: Sixty male rats weighing approximately 200-250g and aged 14-16 wk were divided into 10 groups: sham (Salin + Formalin 1% intraplantar), three treatment groups with lemon essential oil (EO) (12.5, 25 and 50 mg/kg), three treatment groups with Chlorpheniramine (5, 10 and 20 mg/kg), 1 treatment group with Histamine (10 mg/kg), 1 pretreatment group with Chlorpheniramine (20 mg/kg) + EO (50mg/kg), and 1 pretreatment group with Histamine (10 mg/kg) + EO (50 mg/kg). Formalin test was used to assess somatic pain. Data analysis was performed using one-way ANOVA. Results: Intraperitoneal injection of lemon essential oil reduced the pain response induced by formalin in both phases (P<0.05). Pretreatment with chlorpheniramine and lemon essential oil enhanced the analgesic response in both phases (P<0.05). Conclusion: Lemon essential oil had analgesic effects, probably caused by the histaminergic system. [ABSTRACT FROM AUTHOR]

Abstract: The objective of this paper was to study the effect of essential oils (EOs) thymol and carvacrol applied in citrus packing lines in combination with commercial wax on microbial spoilage, lemon decay and quality parameters during postharvest storage and compared with control (tap water), wax and wax + imazalil as positive control. Results showed that the treatment with wax + EOs was able to reduce yeast and mould and total aerobe counts on lemon surface and decayed lemons after 8 weeks of storage, the efficacy being similar to that obtained from wax + imazalil treatment. In addition, lemons coated with EOs or imazalil showed similar quality attributes (weight loss, softening and colour) retention compared with control or waxed fruits. [Copyright &y& Elsevier]

Highlights: [•] Characterisation of phenolic compounds in whole-lemon powder by HPLC–ESI-TOF-MS. [•] Study of the stability under different drying processes and storage conditions. [•] Determination of furanic compounds to evaluate heat damage. [•] Low temperature drying negatively influences the phenolic composition of lemon. [•] Heat damage could be controlled by phenolic profile and antioxidant activity. [ABSTRACT FROM AUTHOR]

Proteins acting as powerful inhibitors of plant pectin methylesterase and polygalacturonase were isolated from whole lemon fruits (Citrus limon L.). Pectin methylesterase inhibitor (PMEI) and polygalacturonase inhibitor protein (PGIP) were purified using DEAE Sepharose column, resulting in fold purity of 89.13 and 81.16 and having a molecular mass of 35 and 38 kDa, respectively as estimated using SDS-PAGE and MALDI-TOF mass spectroscopy. The optimum pH of purified PMEI and PGIP was pH 6 and pH 4.5 while the inhibitors showed good stability in the pH range of 5–8 and 3.5 to 5.5, respectively. Both the inhibitors from C. limon demonstrated an optimum temperature of 55 °C. Thermal inactivation data suggested that purified PGIP was more heat stable than PMEI. The inhibition kinetics of PMEI and PGIP towards C. limon PME and C. limon PG was of a non-competitive type. Both PMEI and PGIP obeyed first-order inactivation kinetics. The PMEI and PGIP exhibited different extent of inhibition towards PME and PG from other fruit sources analyzed in this study. As these inhibitors inhibit PME and PG from other plant sources they can be used in fruit-based products to control undesirable endogenous enzyme activities as an alternative to thermal processing. [Display omitted] • Citrus limon PMEI was purified up to 89.13 fold using DEAE Sepharose resin. • PMEI (∼35 kDa) showed optimum inhibitory activity at 55 °C and pH 6.0 • Citrus limon PGIP was purified to 81.16 fold using DEAE Sepharose resin. • PGIP ∼38 kDa showed optimum inhibitory activity at 55 °C and pH 4.5 • PGIP was more heat stable than PMEI. [ABSTRACT FROM AUTHOR]

Brown rot is a common and devastating disease of different fruits and vegetables. During 2018, a profound brown rot was observed on lemon (Citrus lemon L.) in Islamabad, Pakistan and adjacent areas. This study was designed to identify disease causing pathogen of this disease. Diseased lemon fruit were collected from the orchards and disease causing pathogen was isolated. The morphology of pathogen was examined under stereoscope and compound microscope and the pathogen was identified as Alternaria alternata. For the molecular characterization of this fungus, 18S ribosomal RNA gene was sequenced and its phylogenetic analyses were performed. BLAST analysis revealed 100% similarity with A. alternate NS8, 18S ribosomal RNA gene, partial sequence (MK346034.1). This is the first report of brown rot of lemon, caused by A. alternata in Pakistan. For ecofriendly control of this disease, extracts of indigenous medicinal plants including Justacia adhatoda, Azadriachta indica, Foeniculum vulgare and Mentha spicata were used. In these plant extracts, the presence of various functional groups such as alcohols, carboxylic acids, esters, alkanes and alkenes was revealed by Fourier transform infrared spectroscopy (FTIR). Both in vitro and in vivo assay showed the significant role of aqueous plant extracts to suppress the growth of A. alternata. Among these phytoextracts, A. indica proved to be the most efficient in suppressing the growth of the pathogen and controlling disease. Findings of this study suggest the use of these plant extracts for environment friendly control of fruit rot disease. • Isolation of Fungi from lemon fruit. • Evaluation of pathogenicity of the isolated fungi. • Control of brown rot disease by using Phytoextracts. [ABSTRACT FROM AUTHOR]

Background: Exosome-like nanovesicles are biological nanostructures mediating cell-tocell communication and capable to load selected cargos also in the interaction among different species., Objective: We aimed to explore the content of exosome-like nanovesicles derived from Citrus limon L. and to analyze the effects of their uptake on human cells., Method: We isolated exosome-like nanovesicles from Citrus limon L. juice (EXO-CLs) by differential centrifugation. EXO-CLs were analyzed for short RNA content by advanced sequencing technologies, and for ascorbic acid (vitamin C) and citrate content by enzymatic assays. EXO-CLs anti-oxidant and pro-differentiative potential was evaluated in vitro on mesenchymal stromal cells (MSC), a common tool for regenerative strategies for several human tissues., Results: We showed that EXO-CLs carry detectable amounts of citrate and vitamin C and, although it was not possible to identify specific miRNAs, we detected short RNA sequences (20-30 bp) with unknown functions and with different distribution size in respect to whole Citrus limon L. juice. In vitro, EXO-CLs were uptaken by MSC and had a significant protective effect against oxidative stress. Furthermore, regarding the potential benefit for human bone health, we found that EXO-CLs modulate MSC differentiation versus the osteogenic lineage., Conclusion: We demonstrated that incubation with EXO-CLs exert antioxidant activity in human cells. This is most likely due to the direct delivery and uptake of micronutrients by human cells that are well preserved inside the nanovesicle membrane, including the unstable vitamin C. Based on our results, we speculate that fruit-derived nanovesicles have the potential to mediate interspecies influence after food intake., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Nanosized vesicles are considered key players in cell to cell communication, thus influencing physiological and pathological processes, including cancer. Nanovesicles have also been found in edible-plants and have shown therapeutic activity in inflammatory bowel diseases; however information on their role in affecting cancer progression is missing.Our study identify for the first time a fraction of vesicles from lemon juice (Citrus limon L.), obtained as a result of different ultracentrifugation, with density ranging from 1,15 to 1,19 g/ml and specific proteomic profile. By using an in vitro approach, we show that isolated nanovesicles inhibit cancer cell proliferation in different tumor cell lines, by activating a TRAIL-mediated apoptotic cell death. Furthermore, we demonstrate that lemon nanovesicles suppress CML tumor growth in vivo by specifically reaching tumor site and by activating TRAIL-mediated apoptotic cell processes. Overall, this study suggests the possible use of plant-edible nanovesicles as a feasible approach in cancer treatment.