Sambucus Sieboldiana Research Articles

In situ characterization of the agglutination of lectins via cross-linking of carbohydrates by time-resolved measurement of forward static light scattering.

We present real-time observations of a structurally variable process for cross-linking agglutination between multivalent lectins and glycoclusters using a small-angle forward static light scattering (F-SLS) technique. In this study, a cross-linking agglutination reaction was carried out using a tetravalent Neu5Acα2,6LacNAc-glycocluster and Sambucus sieboldiana agglutinin (SSA). The scattering intensity of time-resolved F-SLS increased with formation of the Neu5Acα2,6LacNAc-glycocluster-SSA cross-linked complex. Using this approach, fine sequential cross-linking agglutination between glycoclusters and lectins was observed in real-time. The rate of increase in the intensity of time-resolved F-SLS increased with the concentration of sialo-glycoclusters and SSA. Structural analysis based on the fractal dimension using time-resolved F-SLS patterns revealed that the density of the aggregates changed with progression of the cross-linking reaction until equilibrium was reached. This is the first report to evaluate the cross-linking agglutination reaction between glycoclusters and lectins and analysis of the subsequent structure of the obtained aggregates using time-resolved measurements of F-SLS.

Changes in Gastric Mucosal Glycosylation Before and After Helicobacter pylori Eradication Using Lectin Microarray Analysis.

Glycosylation is a common post-translational modification, and it has been reported that alterations in the glycosylation patterns on cells are related to cell proliferation, differentiation, tissue adhesion, and carcinogenesis. This study aimed to investigate the relationship between Helicobacter pylori infection and gastric mucosal glycosylation using a lectin microarray system. Gastric mucosal samples were obtained from 10 Helicobacter pylori-non-infected patients, 10 H. pylori-infected patients, and 10 after H. pylori-eradicated patients who underwent gastric mucosal biopsy by endoscopy in our institute. The gastric gland cells which were isolated from formalin-fixed, paraffin-embedded gastric mucosal biopsy samples using laser capture microdissection were used for lectin microarray to obtain lectin-glycan interaction values. Comparison of the lectin-glycan interaction values before and after eradication in the same patients showed significant increases for Ricinus communis agglutinin 120, Trichosanthes japonica agglutinin II, Euonymus europaeus lectin, jacalin, Amaranthus caudatus agglutinin, and Maclura pomifera agglutinin and significant decreases for Urtica dioica agglutinin, Lycopersicon esculentum lectin, Ulex europaeus agglutinin, Sambucus nigra agglutinin, Sambucus sieboldiana agglutinin, and Trichosanthes japonica agglutinin I. Furthermore, jacalin and MPA in the gastric antrum were significantly decreased with H. pylori infection compared with the without infection group and improved to the levels seen without infection as a result of eradication. Lycopersicon esculentum lectin, Sambucus nigra agglutinin, Sambucus sieboldiana agglutinin, and Trichosanthes japonica agglutinin I in the gastric body were significantly increased with H. pylori infection and improved to the level seen without infection as a result of eradication. H. pylori infection changes the lectin binding state which is related to various cancers on the gastric mucosal cell. Furthermore, those changes are reversible by H. pylori eradication.

Retraction to: Inhibitory activity against biological enzyme and anti-microbial activity of phenolics fromSambucus sieboldianavar.pendulaLeaves

Retraction to: Inhibitory activity against biological enzyme and anti-microbial activity of phenolics from<i>Sambucus sieboldiana</i>var.<i>pendula</i>Leaves

Inhibitory activity against biological enzyme and anti-microbial activity of phenolics from Sambucus sieboldiana var. pendula Leaves

말오줌나무 잎 추출물의 생리활성 및 항균활성을 측정한 결과, 말오줌나무 잎 추출물의 생리활성은 말오줌나무 잎 추출물에 함유되어있는 phenolic성분들에 의해 효과가 발현되는 것으로 판단되었다. 말오줌나무 잎으로부터 생리활성에 관여하는 phenolic 화합물을 물과 에탄올로 추출하였을 때 각각 11.60±0.18과 12.39±0.07 mg/g의 함량을 나타내었다. 말오줌나무 잎의 ACE 저해 활성은 50-200 μg/mL의 phenolic을 함유한 물과 에탄올 추출물에서 각각 71.94-92.08와 48.42-78.33%의 저해 활성이 측정되었으며, 농도의존적인 양상을 나타내었다. 말오줌나무 잎 추출물의 XOase에 대한 추출물의 저해 활성은 50-200 μg/mL의 phenolic을 함유한 물과 에탄올 추출물에서 각각 42.11-100와 35.61-100%의 매우 우수한 저해효과를 나타내었다. 말오줌나무잎 추출물에 대한 HAase 저해활성은 200 μg/mL의 phenolic을 함유한 농도에서 물과 에탄올 추출물이 각각 25.35±0.41와 43.38±2.26%의 HAase 저해활성을 나타내었다. 말오줌나무 잎추출물의 항균활성은 충치균인 S. mutans와 식중독균인 S. aureus 및 여드름균인 P. acnes에 대한 우수한 항균 효과를 나타내었다. 따라서 말오줌나무는 고혈압억제, 통풍억제 및 염증억제 작용 등의 생리활성 및 S. mutans와 S. aureus 및 P. acnes에 대한 항균활성이 우수한 것으로 확인되어 말오줌나무를 활용한 기능성 식품 소재로서 산업화에 활용할 수 있을 것으로 생각되었다.

Species-specific and heterogeneous distribution of sialoglycoconjugates in the primary olfactory center of three species of Asian salamanders (Cynops)

Sialic acids (Sia) are terminal components of glycoconjugates that are involved in molecular and cellular interactions in the olfactory system. Diverse glycoconjugates are expressed in the salamander olfactory projection; however, their sialylation and the linkage of Sia to underlying sugars remain largely unknown. The present study aimed to determine the expression of Sia linked to galactose (Gal)-N-acetylglucosamine and N-acetylgalactosamine (GalNAc) in the olfactory bulbs of three species of salamanders using lectin binding. Abundant distribution of sialoglycoconjugates was observed in the salamander olfactory bulb by lectins, Sambucus sieboldiana (SSA) and Maackia amurensis (MAM). Moreover, SSA and MAM showed heterogeneous bindings in the primary olfactory projection of Cynops pyrrhogaster and C. orientalis. Lectin reactivities obviously decreased in all layers of the olfactory bulb after sialidase digestion, indicating selective binding to sialoglycoconjugates. Next, we examined the expression of the subterminal sugar residues, Gal and GalNAc, after terminal Sia removal. Desialylation in the olfactory bulb enhanced the reactivity of Jacalin and Vicia villosa (VVA) lectins that recognize Gal and GalNAc respectively. Together with the binding of SSA and MAM, Sia linked to Gal and GalNAc might be a major component of sialoglycoconjugates in the salamander olfactory projection.

Aqueous synthesis of sialylglycopeptide‐grafted glycopolymers with high affinity for the lectin and the influenza virus hemagglutinin

ABSTRACTGlycopolymers with pendant complex‐type sialyl N‐glycans containing heptapeptides, that is, sialylglycopeptides (SGPs), were synthesized using a water soluble polymer backbone bearing N‐hydroxysulfosuccinimidyl esters by post‐polymerization modification in water. Although SGP has three amino groups on the peptide chain, the substitution reaction occurs preferentially at the N‐terminus α‐amino group in the lysine residue onto the polymer side chain because the reactivity of such α‐amino group is higher than that of the ε‐amino group in the lysine residue under mild acidic aqueous condition. The resulting SGP‐grafted glycopolymers exhibited strong interaction with the lectin Sambucus sieboldiana agglutinin and the human influenza A virus hemagglutinin, with higher binding associate constant values than those of free saccharide according to quartz crystal microbalance analysis. © 2020 Wiley Periodicals, Inc. J. Polym. Sci. 2020, 58, 548–556

Simple and Rapid Detection of Glycoforms by "Lectin Inhibition" Assay.

Glycoforms are otherwise identical proteins with different glycosylation. A lectin, Sambucus sieboldiana agglutinin (SSA), specifically binds glycoforms having α2,6-sialyl residues. The binding is found to inhibit antigen-antibody reaction; e.g., SSA inhibits anti-transferrin antibody binding to α2,6-sialylated transferrin (Tf) (SSA inhibition). SSA inhibition is not observed with other Tf glycoforms, indicating that the inhibition is glycoform-specific. Here we describe the application of SSA inhibition to ELISA as a specific assay for quantifying α2,6-sialylated Tf.

Lectin-Based Assay for Glycoform-Specific Detection of α2,6-sialylated Transferrin and Carcinoembryonic Antigen in Tissue and Body Fluid.

Antibodies are useful for detecting glycoprotein antigens, but a conventional antibody recognizes only a protein epitope rather than a glycan. Thus, glycan isoform detection generally requires time- and labor-consuming processes such as lectin affinity column chromatography followed by sandwich ELISA. We recently found antigen-antibody reactions that were inhibited by lectin binding to glycans on the glycoprotein antigen, leading to a convenient glycoform-specific assay. Indeed, Sambucus sieboldiana agglutinin (SSA) lectin, a binder to sialylα2,6galactose residue, inhibited antibody binding to α2,6-sialylated transferrin (Tf) (SSA inhibition). SSA inhibition was not observed with other glycoforms, such as periodate-treated, sialidase-treated and sialidase/galactosidase-treated Tf, suggesting that the assay was glycoform-specific. SSA inhibition was also applicable for visualizing localization of α2,6-sialylated-Tf in a liver section. This is the first immunohistochemical demonstration of glycoform localization in a tissue section. SSA inhibition was utilized for establishing ELISA to quantify α2,6-sialylated carcinoembryonic antigen (CEA), a marker for various cancers. In addition, α2,6-sialylated-CEA was visualized in a colonic adenocarcinoma section by SSA inhibition. The method would further be applicable to a simple and rapid estimation of other α2,6-sialylated glycoproteins and have a potential aid to histopathological diagnosis.

Two non-cytotoxic type 2 ribosome-inactivating proteins (Sambucus Sieboldiana lectin and Sambucus Nigra lectin) lead neurosphere cells to caspase-independent apoptosis

We found a new phenomenon of cell damage and/or cell death of almost all types of neurosphere cells derived from telencephalons of mouse embryos. The cell damage/death was induced by non-cytotoxic type 2 ribosome-inactivating proteins: Sambucus Sieboldiana lectin (SSA) and Sambucus Nigra lectin (SNA). Self-renewal and aggregation of the neurosphere cells were inhibited under the presence of SSA or SNA. These lectins led the neurosphere cells to cell damage by making them lose their proliferation capacity. It was found that Siaα2-6Galβ1-4GlcNAc or Siaα2-6Gal on the membrane of neurosphere cells play an important role in the induction of cell damage of neurosphere cells. Moreover, despite the fact that most of the neurosphere cells cultured under the presence of SSA or SNA were Annexin V positive, these cells were not identified by DNA ladder assays. Additionally, the cells were not rescued from cell death even under the presence of a pan-caspase inhibitor. These findings imply that cell death of neurosphere cells caused by SSA or SNA is caspase-independent apoptosis via an exogenous pathway. SSA and SNA recognition molecules were detected as 39 and 42 kDa glycoproteins on the membrane of neurosphere cells.

Constituents from the Branches of Sambucus sieboldiana var. pendula with the Properties of Collagen Synthesis Activation

Article history: Received on: 02/02/2015 Revised on: 12/02/2015 Accepted on: 21/03/2015 Available online: 27/04/2015 For the purpose of developing anti-wrinkle cosmetic ingredients, the extracts from branches of a woody plant Sambucus sieboldiana var. pendula were examined on collagen synthesis activities using fibroblast HDFn cells. As a result, the S. sieboldiana ethanol extract (SSE) proved to activate the production of type I procollagen in a dose-dependent manner without showing cell toxicity. Phytochemical study was conducted to isolate the active constituents in the extracts by solvent fractionation followed by chromatographic purifications. From this procedure, two known compounds, kaempferol 3-O-sophoroside (1) and daucosterol (2), were identified by spectroscopic studies. From the isolates, the flavonoid glycoside 1 was verified to induce the synthesis of the type I procollagen dose-dependently. These results suggested that S. sieboldiana extract containing the flavonoid 1 could be useful as an active ingredient in wrinkle-care cosmetics.

In situ visualization of a glycoform of transferrin: localization of α2,6-sialylated transferrin in the liver.

We previously found that a lectin, Sambucus sieboldiana agglutinin (SSA), bound to α2,6-sialylated glycan epitopes on transferrin and inhibited anti-transferrin antibody binding to the antigen in ELISA (SSA inhibition). Here we report that SSA inhibition is applicable to immunohistochemistry, localizing α2,6-sialylated transferrin in the liver. Immunohistochemistry using anti-transferrin polyclonal antibody revealed that transferrin was detected in hepatocytes near interlobular veins. Addition of SSA lectin markedly attenuated the staining. Sialidase treatment of a liver section abolished SSA binding and concomitantly cancelled SSA inhibition, suggesting that SSA binding to glycan epitopes on the section was essential for the inhibition. To examine the importance of proximity between antigen epitopes and SSA-binding (glycosylation) sites, we prepared two anti-peptide antibodies against partial amino acid sequences of transferrin. One antibody (Tf-596Ab) is against a peptide sequence, Cys596-Ala614, which is proximal to N-glycosylation sites (Asn-432 and Asn-630). The other (Tf-120Ab) is against a peptide sequence, Val120-Cys137, distal to the sites. The staining signals of Tf-596Ab were reduced by the addition of SSA, whereas those of Tf-120Ab were reduced only a little. This result suggests that proximity of the antigen epitope to SSA binding sites is critical for SSA inhibition in immunohistochemistry.

Two cases of placental mesenchymal dysplasia: Case reports

Two cases of placental mesenchymal dysplasia: Case reports

Effect of Sambucus sieboldiana Extract on the Cell Growth and Extracellular Matrix Formation in Osteoblast Cells

Sambucus sieboldiana (SS) is a member of the family Caprifoliaceae and has been recommended as a functional material because of its several bioactivities. Although numerous literatures are available on the pharmacological and biological activities, the biological activity of SS in bone regeneration process has not yet been well-defined. Therefore, in this study, the effect of SS was investigated in the proliferation and differentiation of MC3T3-E1 osteoblastic cell line. The treatment of SS did not significantly affect the cell proliferation in MC3T3-E1 cells. SS significantly accelerated the mineralization and significantly increased the expression of alkaline phosphatase (ALP) and osteocalcin (OC) mRNAs, compared to the control, in the differentiation of MC3T3-E1 cells. SS significantly accelerated the decrease of osteonectin (ON) mRNA expression as compared with the control in a time-dependent manner in the differentiation of MC3T3-E1 cells. These results suggest that the SS facilitate the osteoblast differentiation and mineralization in MC3T3-E1 osteoblastic cells. Therefore, there may be potential properties for development and clinical application of bone regeneration materials.

The Distribution of Vascular Plants in Ulleungdo and Nearby Island Regions (Gwaneumdo, Jukdo), Korea

The distribution of vascular plants in Ulleungdo Island and nearby islands (Gwaneumdo and Jukdo) located in Ulleung-gun area, Gyeongsangbuk-do. On-site investigation was conducted twice on June and September, 2012, investigating total 99 families, 304 genera and 508 taxa. This study also arranged already-secured specimens of the Korea National Arboretum (KH) (1917-2011), organizing 106 families, 362 genera and 646 taxa. As a result, total 108 families, 375 genera and 685 taxa of vascular plants were organized in the investigation area. In detail, this study confirmed 30 taxa of endemic plants including Sambucus sieboldiana var. pendula, Silene takeshimensis, and Corydalis filistipes, 41 taxa of rare plants including Arisaema takesimense, Dendranthema zawadskii var. lucidum, and Tripterospermum japonicum, 4 taxa of special plants of forest protection designated by the Korea Forest Service including Abelia coreana var. insularis, Fagus engleriana. Besides, specific plants designated by the Ministry of Environment were 121 taxa, and naturalized plants were 89 taxa.

A lectin microarray study of glycoantigens in neonatal porcine islet-like cell clusters

BackgroundBesides α-Gal expression, the differences of glycosylation and antigenicity between adult pig islets (APIs) and neonatal porcine islet-like cell clusters (NPCCs) are altogether unclear. In this study, lectin microarray analyses of NPCCs were performed and the results compared with the corresponding values for wild-type APIs and NPCCs from α-Gal transferase knockout (GalT-KO) pig. MethodsNPCCs were isolated from 1–3-d-old neonatal wild-type pigs and cultured for 1 d, 5 d, and 9 d, using a previously described technique. Alternatively, the isoration of APIs were isolated based on the method for human islets. ResultsIn a comparison between NPCCs and APIs, all of the NPCCs showed higher signals for Sambucus nigra, Sambucus sieboldiana, and Trichosanthes japonica I and the binding of α2,6 sialc acid, whereas the APIs showed stronger signals for Lotus tetragonolobus, Aleuria aurantia, Narcissus pseudonarcissus, and Galanthus nivalis, suggesting that APIs contain high levels of high-mannose forms. Among the NPCCs, NPCC (day1) appeared to be richer than the others in Lotus tetragonolobus, Narcissus pseudonarcissus, Galanthus nivalis, and Urtica dioica, implying the presence of high-mannose forms. However, as a whole, the signals for many lectins for NPCCs were very similar. The NPCCs from a GalT-KO pig indicated not only the downregulation of α-Gal expression but α-GalNAc as well, and α2-6 sialic acid was upregulated. ConclusionsThe results reported herein contain useful information for the future production of immunomodified pigs with less antigenicity than GalT-KO pigs toward clinical applications of NPCCs.

Glycan profiling of endometrial cancers using lectin microarray

Cell surface glycans change during the process of malignant transformation. To characterize and distinguish endometrial cancer and endometrium, we performed glycan profiling using an emerging modern technology, lectin microarray analysis. The three cell lines, two from endometrial cancers [well-differentiated type (G1) and poorly differentiated type (G3)] and one from normal endometrium, were successfully categorized into three independent groups by 45 lectins. Furthermore, in cancer cells, a clear difference between G1 and G3 type was observed for the glycans recognized with six lectins, Ulex europaeus agglutinin I (UEA-I), Sambucus sieboldiana agglutinin (SSA), Sambucus nigra agglutinin (SNA), Trichosanthes japonica agglutinin I (TJA-I), Amaranthus caudatus agglutinin (ACA), and Bauhinia purpurea lectin (BPL). The lectin microarray analysis using G3 type tissues demonstrated that stage I and stage III or IV were distinguished depending on signal pattern of three lectins, Dolichos biflorus agglutinin (DBA), BPL, and ACA. In addition, the analysis of the glycans on the ovarian cancer cells showed that only anticancer drug-sensitive cell lines had almost no activities to specific three lectins. Glycan profiling by the lectin microarray may be used to assess the characteristics of tumors and potentially to predict the success of chemotherapy treatment.

Associated expression of α2,3sialylated type 2 chain structures with lymph node metastasis in distal colorectal cancer

One of the typical modifications on the surface of cancer cells is sialylation of terminal carbohydrates. The expression of several types of sialylation of glycoconjugates was investigated in colorectal cancer. The cancer tissue specimens obtained from 65 colorectal cancer patients were stained with sialic acid-binding lectins from Maackia amurensis (MAM), Sambucus sieboldiana (SSA), Maackia amurensis agglutinin (MAA) and monoclonal antibodies, and compared with their clinicopathological features. Cancer tissue specimens from 44.6% of patients had positive staining with MAM, which recognized α2,3sialylated type 2 chain (NeuAcα2,3Galβ1,4GlcNAcβR) structures, but normal colorectal mucosa showed only weak staining with MAM was observed. More lymph node metastases and lymphatic invasion were seen in patients with positive staining with MAM (P < 0.01), while not with other lectins or antibodies that recognized sialylated glycoconjugates or sialyl Lewis-related antigens. The five-year survival rate of patients with MAM-positive staining was significantly lower than that with MAM-negative staining when including T0-1 cases, but there was no difference in cases with T2-4. There was no difference in the patients' survival rates when the tissues were stained with MAA, SSA or PNA lectins. α2,3Sialylated type 2 chain structures were predominantly expressed in colorectal tissues associated with the malignant transformation, in particular, with lymphatic spread of distal colorectal adenocarcinomas.

Design and Synthesis of High-Avidity Tetravalent Glycoclusters as Probes for Sambucus sieboldiana Agglutinin and Characterization of their Binding Properties

We designed and synthesized three tetravalent sialo-glycoclusters that had different separations between the terminal sialic acids and the linking carboxy groups of the ethylene glycol bis(β-aminoethyl ether)-N,N,N',N'-tetraacetate scaffold to serve as ligands for the sialic acid-binding lectin Sambucus sieboldiana agglutinin (SSA). The interaction between each glycocluster and SSA was characterized by hemagglutination inhibition, quantitative precipitation, and double-diffusion assays. For the precipitation assays, the precipitin curves indicated that the ligands and SSA bound in either a 1:1 or a 1:2 ratio, i.e., stoichiometrically. The strong interactions of these sialo-glycoclusters with SSA could be ascribed to a combination of multivalency and spacer effects. We also assessed the nature of the ligand-SSA complexes by isothermal titration calorimetry and dynamic light scattering. The results of those experiments indicated that formation of intermolecular complexes occurred at less than stoichiometric ratios of ligand to SSA concentrations and that, as the concentrations of the ligands increased, larger cross-linked aggregates formed. Large aggregates that were present concurrently with visible precipitation and with a particle size centered at ~600 to 800 nm were identified by dynamic light scattering.

Identification of Glycan Structure Alterations on Cell Membrane Proteins in Desoxyepothilone B Resistant Leukemia Cells

Resistance to tubulin-binding agents used in cancer is often multifactorial and can include changes in drug accumulation and modified expression of tubulin isotypes. Glycans on cell membrane proteins play important roles in many cellular processes such as recognition and apoptosis, and this study investigated whether changes to the glycan structures on cell membrane proteins occur when cells become resistant to drugs. Specifically, we investigated the alteration of glycan structures on the cell membrane proteins of human T-cell acute lymphoblastic leukemia (CEM) cells that were selected for resistance to desoxyepothilone B (CEM/dEpoB). The glycan profile of the cell membrane glycoproteins was obtained by sequential release of N- and O-glycans from cell membrane fraction dotted onto polyvinylidene difluoride membrane with PNGase F and β-elimination respectively. The released glycan alditols were analyzed by liquid chromatography (graphitized carbon)-electrospray ionization tandem MS. The major N-glycan on CEM cell was the core fucosylated α2-6 monosialo-biantennary structure. Resistant CEM/dEpoB cells had a significant decrease of α2-6 linked sialic acid on N-glycans. The lower α2-6 sialylation was caused by a decrease in activity of β-galactoside α2-6 sialyltransferase (ST6Gal), and decreased expression of the mRNA. It is clear that the membrane glycosylation of leukemia cells changes during acquired resistance to dEpoB drugs and that this change occurs globally on all cell membrane glycoproteins. This is the first identification of a specific glycan modification on the surface of drug resistant cells and the mechanism of this downstream effect on microtubule targeting drugs may offer a route to new interventions to overcome drug resistance.

Partial-filling affinity capillary electrophoresis of glycoprotein oligosaccharides derivatized with 8-aminopyrene-1,3,6-trisulfonic acid

Partial-filling affinity capillary electrophoresis has been applied to the simultaneous analysis of interactions between glycoprotein oligosaccharides and certain plant lectins. A lectin solution and a mixture of glycoprotein-derived oligosaccharides labeled with 8-aminopyrene-1,3,6-trisulfonic acid were introduced to a neutrally coated capillary in this order, and separated by application of a negative voltage. Interaction of a lectin with each oligosaccharide in the mixture was observed as the specific retardation or dissipation of peaks, in addition to the size/charge separation of oligosaccharides by zone electrophoresis in the remainder (≈90%) of the capillary. The strength of the interaction with lectin was controlled by introducing an appropriate volume of lectin solution. Application of various specificities of lectins indicated characteristic migration profiles of the oligosaccharides. Moreover, sequential injection of four lectins ( Maachia amurensis mitogen, Sambucus sieboldiana agglutinin, Erythrina cristagalli agglutinin, Aleuria aurantia lectin) induced complete dissipation of complex-type oligosaccharides and enabled specific determination of the presence of high-mannose oligosaccharides without the interference or alteration of the electropherogram in porcine thyroglobulin. This method was also applied to determine the binding constants of ovalbumin-derived oligosaccharides to wheat germ agglutinin.