Characterization of mineral composition of leaves and flowers of wild-growing Sambucus nigra.

The objective of this study was to determine the mineral content in the leaves and flowers of wild-grown Sambucus nigra collected from eleven different locations in Kosovo. The samples were digested in a microwave system using the wet digestion method. The minerals were determined by the application of inductively coupled plasma-atomic emission spectrometry (ICP-AES) and inductively coupled plasma-mass spectrometry (ICP-MS). A total of 31 elements were determined, 15 elements by the ICP-AES method (Al, B, Ba, Ca, Cr, Cu, Fe, K, Mg, Mn, Na, P, Sr, V, and Zn) and 16 elements by the ICP-MS method (Ag, As, Be, Bi, Cd, Co, Cs, Ga, Hg, In, Li, Ni, Pb, Rb, Tl, and U). The leaves of S. nigra show a higher content of minerals compared to the flowers, except for the flower of the sample SN-FL10, which is characterized by a high concentration of Fe, Al, Pb, Be, and Tl. The concentration of heavy metals and toxic elements (Pb, Cd, and Hg) was within the permissible concentrations according to Eur. Ph.

Elder (Sambucus nigra), identified by high-content screening, counteracts foam cell formation without promoting hepatic lipogenesis.

Cholesterol deposition in intimal macrophages leads to foam cell formation and atherosclerosis. Reverse cholesterol transport (RCT), initiated by efflux of excess cholesterol from foam cells, counteracts atherosclerosis. However, targeting RCT by enhancing cholesterol efflux was so far accompanied by adverse hepatic lipogenesis. Here, we aimed to identify novel natural enhancers of macrophage cholesterol efflux suitable for the prevention of atherosclerosis. Plant extracts of an open-access library were screened for their capacity to increase cholesterol efflux in RAW264.7 macrophages trace-labeled with fluorescent BODIPY-cholesterol. Incremental functional validation of hits yielded two final extracts, elder (Sambucus nigra) and bitter orange (Citrus aurantium L.) that induced ATP binding cassette transporter A1 (ABCA1) expression and reduced cholesteryl ester accumulation in aggregated LDL-induced foam cells. Aqueous elder extracts were subsequently prepared in-house and both, flower and leaf extracts increased ABCA1 mRNA and protein expression in human THP-1 macrophages, while lipogenic gene expression in hepatocyte-derived cells was not induced. Chlorogenic acid isomers and the quercetin glycoside rutin were identified as the main polyphenols in elder extracts with putative biological action. In summary, elder flower and leaf extracts increase macrophage ABCA1 expression and reduce foam cell formation without adversely affecting hepatic lipogenesis.

Detection of Sialic Acid to Differentiate Cervical Cancer Cell Lines Using a Sambucus nigra Lectin Biosensor.

Pap smear screening is a widespread technique used to detect premalignant lesions of cervical cancer (CC); however, it lacks sensitivity, leading to identifying biomarkers that improve early diagnosis sensitivity. A characteristic of cancer is the aberrant sialylation that involves the abnormal expression of α2,6 sialic acid, a specific carbohydrate linked to glycoproteins and glycolipids on the cell surface, which has been reported in premalignant CC lesions. This work aimed to develop a method to differentiate CC cell lines and primary fibroblasts using a novel lectin-based biosensor to detect α2,6 sialic acid based on attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and chemometric. The biosensor was developed by conjugating gold nanoparticles (AuNPs) with 5 µg of Sambucus nigra (SNA) lectin as the biorecognition element. Sialic acid detection was associated with the signal amplification in the 1500-1350 cm-1 region observed by the surface-enhanced infrared absorption spectroscopy (SEIRA) effect from ATR-FTIR results. This region was further analyzed for the clustering of samples by applying principal component analysis (PCA) and confidence ellipses at a 95% interval. This work demonstrates the feasibility of employing SNA biosensors to discriminate between tumoral and non-tumoral cells, that have the potential for the early detection of premalignant lesions of CC.

Levels and sources of polycyclic aromatic hydrocarbons (PAHs) near hospitals and schools using leaves and barks of Sambucus nigra and Acacia melanoxylon.

Polycyclic aromatic hydrocarbons (PAHs) are among the most studied organic compounds in urban environments, due to their known threat to human health. This study extends the current knowledge regarding the ability of different vegetative parts of different tree species to accumulate PAHs. Moreover, exposure intensity to PAHs in areas frequented by population susceptible to adverse health effects of air pollution is evaluated. For this, leaves and barks of Sambucus nigra (S. nigra) and Acacia melanoxylon (A. melanoxylon) were collected at urban areas in the Andean city of Quito, at seven points near hospitals and schools. A methodology, previously developed, for the extraction, purification, and quantification of PAHs associated with the leaves and bark of S. nigra was employed and also validated for leaves and bark of A. melanoxylon. The total PAH level varied from 119.65 ng g-1 DW (dry weight) to 1969.98 ng g-1 DW (dry weight) with naphthalene (Naph), fluoranthene (Flt), pyrene (Pyr), chrysene (Chry), and benzo[a]pyrene (BaP) predominating in all samples. The results indicate that the leaves and bark of tree species studied have certain abilities to bio-accumulate PAH according to their molecular weight. The leaves of S. nigra and bark of A. melanoxylon showed the highest ability to accumulate PAHs, mainly those with high and medium molecular weight, respectively. The highest incidence of light molecular weight PAHs was found in the leaves of A. melanoxylon. Furthermore, coal combustion, biomass burning, and vehicle emissions were identified as the main PAHs sources. Concentrations of PAHs associated with tree species suggest an affectation in areas frequented by populations susceptible to air pollution. This fact shows the importance of regulatory scheme to significantly improve the air quality in the city integrating a knowledge-based decision-making.

From the name to the popular image of the plant: the Polish names for the black elder (Sambucus nigra).

The names of plants convey information on their appearance (shape, structure, colour), taste or smell, their uses (practical, ceremonial, magical and medicinal) as well as the beliefs and convictions associated with them. Assuming that the particular features of plants, entrenched in their names, must have been important to language users for some reason, the analysis of plant names can help reconstruct traditional knowledge about plants. The author analyses the standard and dialectal names for the black elder (Sambucus nigra) in Polish, juxtaposing the plant's features revealed in its names (linguistic data) with the cultural accounts associated with the plant ("with-linguistic" data). This allows for the reconstruction of the following features of the plant: (a) the appearance of the shrub, (b) the properties of its fruit, (c) the smell of the plant, (d) the place where it grows, (e) the time of harvesting, (f) its use in folk medicine and (g) the association of the plant with impure powers and diseases. The conducted analyses show that reaching for hard "linguistic evidence" (standard and folk names) makes it possible to compile hierarchies of the characteristics of the plants described. Situating these names against the background of "with-linguistic" data leads to the conclusion that folk nomenclature and folk knowledge enrich and complement each other. The vast number of names for the black elder with different onomasiological bases, presenting different points of view, also demonstrate the relationship between the degree of lexical differentiation and the cultural meaning of the plant. The ethnolinguistic analysis of the names for the black elder (Sambucus nigra), similarly to ethnobotanical studies of folk plant names, provides insights into past and contemporary uses of the plant. Thus, it can provide a starting point for further ethnobotanical research.

New esters from the essential oil of dry flowers of elder (Sambucus nigra L.).

BACKGROUND: Elder (Sambucus nigra L.) has relevance for the food, fragrance and pharmaceutical industries. Flowers of this species emit a very pleasant scent; for processing purposes, inflorescences are either collected from the wild or harvested from a cultivated crop. The study of elderflower-derived volatiles bears both phytochemical and commercial importance. RESULTS: Three samples of dry elderflower essential oil obtained from laboratory-scale hydrodistillations were analyzed. By use of gas chromatography-mass spectrometry, synthesis and NMR studies of chromatographic fractions of a distillation water extract prepared in a semi-industrial scale steam distillation, 252 constituents of the oil were identified; 115 compounds were not previously reported as elderflower volatiles, seven of which were new natural esters. Particularly interesting were those of isosenecioic (3-methylbut-3-enoic) acid because these were never before found in the plant kingdom. CONCLUSION: With these identifications, the known essential oil constituents accounted for 89.0-93.0% of the analyzed samples. Although the number of known S. nigra flower-derived volatiles is now quite high, further research (both analytical and olfactory) is needed to unveil all of the relevant contributions to the unique odor of elderflowers. © 2023 Society of Chemical Industry.

Flavonoid-Rich Sambucus nigra Berry Extract Enhances Nrf2/HO-1 Signaling Pathway Activation and Exerts Antiulcerative Effects In Vivo.

Sambucus nigra (SN) berry extract is characterized by high antioxidant and anti-inflammatory activity. The current study aimed to investigate the effect of SN berry extract against indomethacin (IND)-induced gastric ulcer in rats and the mechanism involved. SN berry extract alleviated IND-induced gastric ulcers, as shown by assessing pathological manifestations in the gastric mucosa. These protective effects are attributed to attenuated oxidative damage to the gastric mucosa, correlated to increased activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), enhanced glutathione (GSH) levels, total antioxidant capacity (TAC), and upregulation of the Nrf2/HO-1 cascade. Moreover, oxidative stress markers, including malondialdehyde (MDA) and total oxidant status (TOS), were downregulated in SN-extract-treated animals. Furthermore, SN berry extract suppressed gastric mucosal inflammation by downregulating interleukin (IL)-33, IL-1ß, IL-6, and tumor necrosis factor-alpha (TNF-α) levels, and attenuating myeloperoxidase (MPO) activity. The protective effects of SN berry extract were similar to those exerted by esomeprazole (ESO), an acid-secretion-suppressive drug. In conclusion, SN berry extract has antiulcerative effects, alleviating oxidative stress and inflammation.

Health-Promoting Properties: Anti-Inflammatory and Anticancer Properties of Sambucus nigra L. Flowers and Fruits.

Sambucus nigra L. has been used for centuries in traditional medicine thanks to its valuable healing properties. The healing properties result from its high content of biologically active compounds, mainly antioxidants, which contribute to its anti-inflammatory and anticancer properties. In our review, we have presented scientific studies evaluating the anti-inflammatory and anticancer effects of extracts and their components from S. nigra L. flowers and fruits. The results of the research show that the effect of antioxidant phytochemicals contained in their composition reduces the level of free radicals and pro-inflammatory cytokines, prevents mutations that increase the risk of cancer development, and inhibits cell proliferation, induction of apoptosis, and changes in intracellular signaling, consequently inhibiting the growth of malignant tumors and the formation of metastases. Flowers and fruits of S. nigra L. are a valuable source of nutraceutical and pharmacological substances that can support prevention and anti-inflammatory and oncological therapy without negative side effects for the patient.

The Effect of Sample Pretreatment on the Anthocyanin Content in Czech Wild Elderberry (Sambucus nigra L.).

This work focusses on the analysis of anthocyanins present in wild Czech elderberries, using spectrophotometric methods and liquid chromatography. Spectrophotometric methods were used to determine antioxidant capacity, total phenolic content, and total anthocyanin content. Further, four major elderberry anthocyanins were determined using reversed-phase high-performance liquid chromatography with isocratic elution of 30% aqueous methanol with 5% of formic acid. All optimised methods were applied to the analysis of extracts prepared from frozen and dried elderberry fruit samples, and the results were evaluated using principal component analysis which clearly divided the samples into individual groups according to the sample pretreatment (frozen and dried samples). The frozen samples reached higher values of antioxidant capacity and total phenolic and anthocyanin contents compared to the dried samples, probably due to the degradation of anthocyanins during the drying process.

Anti-Inflammatory and Protective Effects of Water Extract and Bioferment from Sambucus nigra Fruit in LPS-Induced Human Skin Fibroblasts.

In this study, an attempt was made to evaluate the antioxidant, anti-inflammatory and protective effects of the Sambucus nigra fruit extract and its ferment obtained by fermentation with kombucha tea fungus. For this purpose, fermented and non-fermented extracts were compared in terms of their chemical composition by the HPLC/ESI-MS chromatographic method. The antioxidant activity of the tested samples was assessed using DPPH and ABTS assays. Cytotoxicity was also determined using Alamar Blue and Neutral Red tests to assess the viability and metabolism of fibroblast and keratinocyte skin cells. Potential anti-aging properties were determined by their ability to inhibit the activity of the metalloproteinases collagenase and elastase. Tests showed that the extract and the ferment have antioxidant properties and stimulate the proliferation of both cell types. The study also assessed the anti-inflammatory activity of the extract and ferment by monitoring levels of the pro-inflammatory interleukins IL-6, IL-1ß, tumor necrosis factor (TNF-α) and anti-inflammatory IL-10 in lipopolysaccharide (LPS)-treated fibroblast cells. The results indicate that both the S. nigra extract and its kombucha ferment can be effective in preventing free-radical-induced cell damage and have positive effects on skin cell health.

Efficacy of traditional herbal formulas on human immunity.

In the present study, we reviewed the efficacy of traditional herbal formulas on human immunity. A literature survey was performed in PubMed, UpToDate, Proquest Central Databases of Kirikkale University, Google and Google Scholar databases from the internet. Search key words were "immune", "immune system", "herbal", "Pelargonium Sidoides", "Echinacea Purpurea", "Sambucus Nigra", "Beta Glucan", "Vitamin C", "Zinc". The immune system is a natural self-defense mechanism made up of cells that assist the body in distinguishing between self and non-self-molecules. All immune system components must be regularly modified in order to keep the body defenses up against the ever-evolving microbes that are constantly looking for new ways to attack the host. A Chinese herbal formulation is a combination of several herbs. The practitioner begins with one or two major substances that are intended to treat the ailment. The reproducibility of the efficacy of herbal medicines is dependent on the consistency of the quality of each unique raw herb. Pelargonium Sidoides, Echinacea Purpurea, Sambucus Nigra, Beta Glucan, Vitamin C, and Zinc are some herbal treatments utilized for their benefits on human immunity. Herbal remedies are undoubtedly valuable in boosting impaired immune function, particularly where damage has occurred due to malnutrition, chronic disease or previous infections. At present, however, an invincible immune system remains firmly in the realm of fantasy.

Antioxidant properties and antibacterial activity of water extracts from Sambucus nigra L. under different conditions.

INTRODUCTION: In folk medicine, dried white flowers of Sambucus nigra L. are used to make infusions, decoctions, and juices. AIM: The present article aims to study and compare the antioxidant activity of aqueous solutions of leaves and flowers of Sambucus nigra L obtained at different exposure times and assess the antibacterial activity of these solutions against Escherichia coli ATCC 8739, Salmonella NCTC 6017, Listeria monocytogenes NCTC 11994, and Staphylococcus aureus ATCC 25093. MATERIALS AND METHODS: We studied the physicochemical properties of aqueous extracts of leaves (fresh) and flowers (fresh and dry) of Sambucus nigra L collected from the Rhodope region of Bulgaria. The samples from Sambucus nigra L were analyzed to determine their total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). The diameters (in millimeters) of the growth inhibition zones of four pathogens were measured, and a comparative assessment of their antibacterial activity was made. RESULTS: The infusions of fresh blossoms and fresh leaves of Sambucus nigra L had the highest antioxidant activity at the total contact time of 30 minutes (82.7 mmol TE/100 ml) and 35 minutes (36.5 mmol TE/100 ml), respectively. The phenol-richest infusions were those made from dried flowers of Sambucus nigra L after a 30-minute contact time (86.7 mg GAE/ml). Of the four pathogens we studied, we found that the extracts affected partially only the pathogenic bacteria of Salmonella. CONCLUSIONS: The highest content of bioactive components was obtained from dried blossoms of Sambucus nigra L. for infusions with a total contact time of 30 minutes and for decoctions at a contact time of 45 minutes.

Interaction of lectin Sambucus nigra with sialylated trisaccharides in the presence of osmolytes. Chronopotentiometric sensing.

Trisaccharides bind to their interaction partners-lectins relatively weakly, which makes detection of their complexes challenging. In this work, we show that an osmolyte presence improves the distinguishing complexes of lectin Sambucus nigra with trisialyllactoses with various binding affinities. The addition of osmolyte, non-binding sugar mannose significantly improved the precision of binding experiments performed using chronopotentiometric stripping at the electrode surface and fluorescence analysis in solution. Osmolytes minimized nonspecific interactions between binding sugar and lectin. Obtained findings can be utilized in any in vitro methods studying interactions of carbohydrates, respectively their conjugates with proteins. The study of carbohydrate interactions appears important since they play essential roles in a variety of biological processes including carcinogenesis.

Bioactive Potential of Elderberry (Sambucus nigra L.): Antioxidant, Antimicrobial Activity, Bioaccessibility and Prebiotic Potential.

Due to its abundance of physiologically active ingredients, one of the oldest medicinal herbs, elderberry (EB) Sambucus nigra L., is beneficial for both therapeutic and dietary purposes. This study determined the bioaccessibility of the phenolic compounds and the prebiotic potential of the polyphenols from freeze-dried EB powder (FDEBP), along with the antioxidant and antimicrobial activities of this extract. The most significant phenolic compounds in black EB are represented by anthocyanins (41.8%), predominating cyanidin-sambubiosides and cyanidin-glucosides (90.1% of the identified anthocyanins). The FRAP assay obtained the highest antioxidant activity value (185 ± 0.18 µmol Fe2+/g DW). The most sensitive to the antimicrobial activity of the extract was proven to be Staphylococcus aureus, and Pseudomonas aeruginosa had the lowest minimum inhibitory concentration of 1.95 mg/mL. To determine the prebiotic potential of the polyphenols, the cell growth of five probiotic strains (Lactobacillus plantarum, L. casei, L. rhamnosus, L. fermentum and Saccharomyces boulardii) was tested. The influence on cell growth was positive for all five probiotic strains used. Overall, the most significant increase (p < 0.05) was recorded at 1.5% FDEBP, on L. casei with a growth index (GI) of 152.44%, very closely followed by GI at 0.5% and 1% concentrations. The stability of the total phenolic compounds through simulated gastronitestinal digestion was increased (93%), and the bioaccessibility was also elevated (75%).

The Effect of the Plant Stabilisation Method on the Composition and Antioxidant Properties of Elderflower (Sambucus nigra L.) Extract.

Elderflower extracts are known to be a source of valuable substances that show a wide spectrum of biological activity, including antibacterial and antiviral properties, which demonstrate a degree of effectiveness against SARS CoV-2. In this work, the influence of fresh inflorescence stabilisation methods (freezing, air drying, and lyophilisation) and extraction parameters on the composition and antioxidant properties of the extracts were studied. Wild elderflower plants growing in the Malopolska Region of Poland were studied. Antioxidant activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl free radical-scavenging ability and ferric-reducing antioxidant power assays. The total phenolic content was determined using the Folin-Ciocalteu method and the phytochemical profile of the extracts was analysed using HPLC. The obtained results showed that the best method for the stabilisation of elderflower was lyophilisation, and the determined optimal maceration parameters were 60% methanol as a solvent and a process time of 1-2 days.

Phytochemical Profiling of Sambucus nigra L. Flower and Leaf Extracts and Their Antimicrobial Potential against Almond Tree Pathogens.

Despite extensive research on the chemical composition of elderberries and their numerous uses in pharmaceutical, beverage, and food production, there is still a lack of knowledge about Sambucus nigra leaves and flowers' antimicrobial activity against plant pathogens. In this study, the phytoconstituents of their aqueous ammonia extracts were first characterized by infrared spectroscopy and gas chromatography-mass spectrometry. The major phytocompounds identified in the flower extract were octyl 2-methylpropanoate; 3,5-dihydroxy-6-methyl-2,3-dihydropyran-4-one; propyl malonic acid; adenine; and 1-methyl-2-piperidinemethanol. Concerning the leaf extract, 1,6-anhydro-ß-D-glucopyranose; oleic acid; 2,1,3-benzothiadiazole; 2,3-dihydro-benzofuran; and 4-((1E)-3-hydroxy-1-propenyl)-2-methoxyphenol and other phenol derivatives were the main constituents. The potential of the extracts to act as bioprotectants was then investigated against three almond tree pathogens: Diaporthe amygdali, Phytophthora megasperma, and Verticillium dahliae. In vitro tests showed higher activity of the flower extract, with EC90 values in the 241-984 µg·mL-1 range (depending on the pathogen) vs. 354-1322 µg·mL-1 for the leaf extract. In addition, the flower extract led to full protection against P. megasperma at a dose of 1875 µg·mL-1 in ex situ tests on artificially-infected excised almond stems. These inhibitory concentrations were lower than those of commercial fungicides. These findings suggest that S. nigra aerial organs may be susceptible to valorization as an alternative to synthetic fungicides for the protection of this important crop.

Effect of Enriching Gingerbread Cookies with Elder (Sambucus nigra L.) Products on Their Phenolic Composition, Antioxidant and Anti-Glycation Properties, and Sensory Acceptance.

Elder products are still underutilized sources of phytochemicals, mainly polyphenols, with extensive pharmacological effects on the human body. In this study, gingerbread cookies covered in chocolate (GC) were enriched with elderflower dry extract (EF) and juice concentrate (EB). The cookies (GC, GCEF, and GCEFEB) and the additives (EF and EB) were analyzed for total phenolic content (TPC), phenolic compound profile, antioxidant capacity (AC), and advanced glycation end products' (AGEs) formation in both the free and bound phenolic fractions. Sensory analysis of the cookies was performed using an effective acceptance test (9-point hedonic scale), and purchase intent was evaluated using a 5-point scale. It was found that the flavonoid content was significantly increased (20-60%) when EF and EB were added to the cookies. Moreover, the EF addition to chocolate-covered GCs enhanced the content of phenolic acids (up to 28%) in the bound phenolic fraction. An increase in the AC values of enriched cookies was found, and the free phenolic fraction differed significantly in this regard. However, inhibition of AGEs by elder products was only observed in the bound phenolic fraction. In addition, EF and EB improved the overall acceptance of the cookies, mostly their taste and texture. Thus, elder products appear to be valuable additives to gingerbread cookies, providing good sensory quality and functional food characteristics.

Effective removal of Cr (VI) ions using low-cost biomass leaves (Sambucus nigra L.) in aqueous solution.

The tannery industries have become an important part of societal growth; however, these processes have produced huge volumes of effluents containing heavy metals, particularly Cr(VI) oxyanions. The study is crucial and cost-effective for reducing the chromium (VI) from industrial wastewater. In order to meet the sustainable development goal (SDG) objective 6.3, the capacity of Sambucus nigra L. to adsorb heavy metal is established with the purpose of eradicating hazardous chemical contamination and reducing pollution. In this study, discontinuous tests were carried out to determine the efficiency of Cr(VI) sorption on leaves of Sambucus nigra L. Adsorption factors such as pH, temperature, adsorbent dosage, and contact time were evaluated. At a dosage of 3 g/L and pH 2, an efficiency of 98.22% was achieved under favorable conditions. The equilibrium and kinetic models that best fitted the experimental data are non-linear Freundlich and; pseudo-second order, and intra-particle diffusion, respectively. The thermodynamic parameters of the adsorption process, including Gibbs free energy (ΔG0), enthalpy (ΔH0), and entropy (ΔS0), were measured at 291, 303, 323, and 343 K, indicating that the phenomena was spontaneous and endothermic. The chemical analyses and surface morphology of the adsorbent were analyzed using SEM (scanning electron microscopy), EDS (energy dispersive spectroscopy), FTIR (Fourier transform infra-red), XRD (X-ray diffraction), and ICP-OES (inductively coupled plasma optical-emission spectroscopy) techniques. The results showed that Sambucus nigra L. has a significant removal efficiency of Cr(VI) in the contaminated solutions, establishing adsorbent as a low cost, readily available, and environmentally friendly and ensuring its potential for industrial usage.

Assessment of the role of anthocyanin standardized elderberry (Sambucus nigra) extract as an immune-stimulating nutraceutical of Nile tilapia, Oreochromis niloticus.

The study of nutraceuticals and their connection to immunity is an expanding field of research. The use of nutraceuticals to alleviate stress and enhance immunity in adverse aquaculture environments have been examined to a certain extent. To elucidate the understanding, we focused on the immunological effect of membrane-separated 13% anthocyanin standardized elderberry (EB) extract with maltodextrin excipient, widely used first-line nutraceuticals to augment the immunity, in aquaculture fish, Nile tilapia. To evaluate the potential of EB-extract, we assessed their capability to enhance lymphocyte proliferation and interleukin-2 production in an in-vitro condition using spleen and thymus lymphocytes. The experiments on spleen and thymus T-cells demonstrated significantly higher T-cell proliferation by EB-extract when lectin mitogen Con A was present as a stimulator. Likewise, our spleen B-cell proliferation result reveals a significant effect of EB-extracts, along with B-cell stimulator non-lectin mitogen LPS. Further, the quantification of IL-2 indicates elevated IL-2 levels when spleen T-cells were cultured with EB-extracts and with Con A present as a stimulator. These suggest that 13% anthocyanin standardized EB-extracts can aggrandize fish cells' cellular and humoral immune responses. With further research, elderberry extracts could be used to supplement commercial feed in aquaculture to reduce stress and stimulate the immune response.

Structure and Biological Properties of Ribosome-Inactivating Proteins and Lectins from Elder (Sambucus nigra L.) Leaves.

Ribosome-inactivating proteins (RIPs) are a group of proteins with rRNA N-glycosylase activity that catalyze the removal of a specific adenine located in the sarcin-ricin loop of the large ribosomal RNA, which leads to the irreversible inhibition of protein synthesis and, consequently, cell death. The case of elderberry (Sambucus nigra L.) is unique, since more than 20 RIPs and related lectins have been isolated and characterized from the flowers, seeds, fruits, and bark of this plant. However, these kinds of proteins have never been isolated from elderberry leaves. In this work, we have purified RIPs and lectins from the leaves of this shrub, studying their main physicochemical characteristics, sequences, and biological properties. In elderberry leaves, we found one type 2 RIP and two related lectins that are specific for galactose, four type 2 RIPs that fail to agglutinate erythrocytes, and one type 1 RIP. Several of these proteins are homologous to others found elsewhere in the plant. The diversity of RIPs and lectins in the different elderberry tissues, and the different biological activities of these proteins, which have a high degree of homology with each other, constitute an excellent source of proteins that are of great interest in diagnostics, experimental therapy, and agriculture.