Sample records for agglutinins

  1. Febrile/cold agglutinins

    ... diagnose certain infections and find the cause of hemolytic anemia (a type of anemia that occurs when red ... or cold agglutinins can help explain why the hemolytic anemia is occurring and direct treatment.

  2. Salivary agglutinin/glycoprotein-340/DMBT1

    Ligtenberg, Antoon J M; Veerman, Enno C I; Nieuw Amerongen, Arie V;


    Salivary agglutinin (SAG), lung glycoprotein-340 (gp-340) and Deleted in Malignant Brain Tumours 1 (DMBT1) are three names for identical proteins encoded by the dmbt1 gene. DMBT1/SAG/gp-340 belongs to the scavenger receptor cysteine-rich (SRCR) superfamily of proteins, a superfamily of secreted o...

  3. Purification and Characterization of Abrus precatorius Agglutinin

    Absar, Nural; Funatsu, Gunki


    Abrus pvecatorius agglutinin (APA) has been purified by a new purification procedure from the seeds of semen jequiriti produced in Bangladesh and Taiwan. The method was accomplished by 33-50% saturation ammonium sulfate fraction from 1% acetic acid extract of the seeds of semen jequiriti using gel filtration on Sephadex G-75 followed by DEAE-cellulose column chromatography. The molecular weight was estimated to be 126,000 and 122,000 by gel filtration on Sephadex G-150 for Bangladesh-APA and ...

  4. Occurrence of granulocyte cytotoxins and agglutinins.

    Hasegawa, T; Bergh, O J; Terasaki, P I; Graw, R G


    Granulocyte cytotoxic activity in sera from over 257 patients was shown to be distinct from HL-A lymphocytoxic activity. Granulocyte cytotoxins occur in approximately 25 per cent of sera from patients having leukemia, 45 per cent with aplastic anemia, 22 per cent with kidney disease on hemodialysis, and 19 per cent of pregnant women. By testing sera on the same panel of cells, the granulocyte cytotoxic activity was shown not to be associated with granulocyte agglutination activity or lymphocytotoxic acitivty. It is likely that granulocyte cytotoxins and granulocyte agglutinins will be useful in transfusion and bone marrow transplantation as a separate tool from the more widely used lymphocyte cytotoxicity reaction. PMID:1129831

  5. Antibody interactions with Ricinus communis agglutinins studied by biolayer interferometry

    Two related agglutinins are present in the seeds of Ricinus communis (castor): ricin, a dichain ribosome-inactivating protein and Ricinus communis agglutinin-1 (RCA-1), a much less toxic hemagglutinin. Because ricin has been used for experimental cancer chemotherapy as well as for intentional poison...

  6. Llama-Derived Single Domain Antibodies Specific for Abrus Agglutinin

    James P. Carney


    Full Text Available Llama derived single domain antibodies (sdAb, the recombinantly expressed variable heavy domains from the unique heavy-chain only antibodies of camelids, were isolated from a library derived from llamas immunized with a commercial abrin toxoid preparation. Abrin is a potent toxin similar to ricin in structure, sequence and mechanism of action. The selected sdAb were evaluated for their ability to bind to commercial abrin as well as abrax (a recombinant abrin A-chain, purified abrin fractions, Abrus agglutinin (a protein related to abrin but with lower toxicity, ricin, and unrelated proteins. Isolated sdAb were also evaluated for their ability to refold after heat denaturation and ability to be used in sandwich assays as both capture and reporter elements. The best binders were specific for the Abrus agglutinin, showing minimal binding to purified abrin fractions or unrelated proteins. These binders had sub nM affinities and regained most of their secondary structure after heating to 95 °C. They functioned well in sandwich assays. Through gel analysis and the behavior of anti-abrin monoclonal antibodies, we determined that the commercial toxoid preparation used for the original immunizations contained a high percentage of Abrus agglutinin, explaining the selection of Abrus agglutinin binders. Used in conjunction with anti-abrin monoclonal and polyclonal antibodies, these reagents can fill a role to discriminate between the highly toxic abrin and the related, but much less toxic, Abrus agglutinin and distinguish between different crude preparations.

  7. Complement activation by salivary agglutinin is secretor status dependent

    S.T.G. Gunput; A.J.M. Ligtenberg; B. Terlouw; M. Brouwer; E.C.I. Veerman; D. Wouters


    After mucosal damage or gingival inflammation, complement proteins leak into the oral cavity and mix with salivary proteins such as salivary agglutinin (SAG/gp-340/DMBT1). This protein is encoded by the gene Deleted in Malignant Brain Tumors 1 (DMBT1), and it aggregates bacteria, viruses and fungi,

  8. Fatal cold agglutinin-induced haemolytic anaemia: a case report

    Reverberi Roberto


    Full Text Available Abstract Introduction Cold agglutinin disease usually develops as a result of the production of a specific immunoglobulin M auto-antibody directed against the I/i and H antigens, precursors of the ABH and Lewis blood group substances, on red blood cells. Autoimmune and lymphoproliferative disorders, Mycoplasma pneumoniae and other infections can be associated with the production of cold agglutinins. In its classic presentation with haemolytic anaemia and Raynaud's syndrome, cold agglutinin disease is usually idiopathic. Several factors play a role in determining the ability of a cold agglutinin to induce a haemolytic anaemia such as antibody concentration and temperature range, in particular the highest temperature at which antibodies interact with red blood cells. Case presentation A 48-year-old Caucasian man presented to our hospital with symptoms of extreme asthenia caused by severe anaemia. The transfusion of red blood cells (O Rh-positive, started as prescribed by the emergency guidelines in force without pre-transfusion tests, induced fatal haemolysis because of the presence of high levels of anti-H antibodies in his blood, that reacted with the large amount of H antigen in universal (0 red blood cells. Conclusion Emergency transfusion of universal red blood cells (0 Rh-positive or negative is usually accepted by the international guidelines in force in emergency departments. In this report we describe a rare complication caused by the very high concentration in the recipient of cold agglutinins and the activation of the complement system, responsible for red blood cell lysis and consequent fatal cardiovascular shock. We conclude that emergency transfusion of universal red blood cells (0 Rh-positive or negative may be dangerous and its risk should be assessed against the risk of delaying transfusion until the pre-transfusion tests are completed.

  9. Inflammatory and anti-inflammatory effects of soybean agglutinin

    Benjamin C.F.; Figueiredo R.C.; Henriques M.G.M.O.; Barja-Fidalgo C.


    Soybean agglutinin (SBA) lectin, a protein present in raw soybean meals, can bind to and be extensively endocytosed by intestinal epithelial cells, being nutritionally toxic for most animals. In the present study we show that SBA (5-200 µg/cavity) injected into different cavities of rats induced a typical inflammatory response characterized by dose-dependent exudation and neutrophil migration 4 h after injection. This effect was blocked by pretreatment with glucocorticoid (0.5 mg/kg) or by co...

  10. Biogenesis and fate of the cell-cell adhesion molecule, agglutinin, during gametogenesis and fertilization of Chlamydomonas reinhardtii

    Fertilization in Chlamydomonas begins with the species-specific recognition and adhesion between gametes of opposite mating types via agglutinin molecules on the flagellar surface. This adhesion generates a cAMP-mediated sexual signal that initiates the subsequent events of call wall release, mating structure activation, and cell fusion. Although flagella of paired gametes remain attached to each other until the zygote forms, the process is dynamic. Engaged agglutinins rapidly become inactivated and turnover, requiring the constant supply of new agglutinins to replace the lost molecules. A population of cell body associated agglutinins has been postulated to the pool of agglutinins recruited during this turnover. Cell body agglutinins, therefore were identified, purified, localized within the cells and compared to flagellar agglutinins. The relationship between these two agglutinin populations was also examined. Cell body agglutinins were biochemically indistinguishable from the flagellar form with respect to their Mr, sedimentation coefficient, and hydrophobicity elution properties. Functionally, however, these molecules were inactive in situ. The calculated surface density of agglutinins in the cell body and flagellar domains was similar and thus could not explain their functional difference, but two domains contiguous and yet distinctive suggested they may be separated by a functional barrier. To test this, a method was developed, using a monoclonal antibody and cycloheximide, that removed the flagellar agglutinins so movement between the domains could be monitored. Mobilization of agglutinins onto the flagella did not occur unless sexual signaling was induced with cAMP and papaverine

  11. Human salivary agglutinin binds to lung surfactant protein-D and is identical with scavenger receptor protein gp-340

    Ligtenberg, T J; Bikker, F J; Groenink, J;


    agglutinin bound in a similar way to Streptococcus mutans and surfactant protein-D. Histochemically, the distribution of gp-340 in the submandibular salivary glands was identical with the agglutinin distribution, as shown in a previous paper [Takano, Bogert, Malamud, Lally and Hand (1991) Anat. Rec. 230, 307......-318]. We conclude that agglutinin is identical with gp-340, and that this molecule interacts with S. mutans and surfactant protein-D....

  12. Cold agglutinin disease associated with mycoplasma infection in an individual with type 2 diabetes: An atypical case

    Chandrama Shrestha; Liu Min; Zhaohui Mo


    Cold Agglutinin Disease is a hemolytic anemia associated with cold reactive autoantibodies. Although the acute form of cold agglutinin disease can be attributed to autoimmune or infectious diseases and lymphoproliferative diseases, it has, to the best of our knowledge, so far,never been reported as secondary to mycoplasma pneumonia in a type 2 diabetic individual. In this paper, we report a case of cold agglutinin disease following mycoplasma pneumonia in a 47-year-old female patient with typ...

  13. Inflammatory and anti-inflammatory effects of soybean agglutinin

    Benjamin C.F.


    Full Text Available Soybean agglutinin (SBA lectin, a protein present in raw soybean meals, can bind to and be extensively endocytosed by intestinal epithelial cells, being nutritionally toxic for most animals. In the present study we show that SBA (5-200 µg/cavity injected into different cavities of rats induced a typical inflammatory response characterized by dose-dependent exudation and neutrophil migration 4 h after injection. This effect was blocked by pretreatment with glucocorticoid (0.5 mg/kg or by co-injection of N-acetyl-galactosamine (100 x [M] lectin, but not of other sugars (100 x [M] lectin, suggesting an inflammatory response related to the lectin activity. Neutrophil accumulation was not dependent on a direct effect of SBA on the macrophage population since the effect was not altered when the number of peritoneal cells was increased or decreased in vivo. On the other hand, SBA showed chemotactic activity for human neutrophils in vitro. A slight increase in mononuclear cells was observed 48 h after ip injection of SBA. Phenotypic analysis of these cells showed an increase in the CD4+/CD8- lymphocyte population that returned to control levels after 15 days, suggesting the development of an immune response. SBA-stimulated macrophages presented an increase in the expression of CD11/CD18 surface molecules and showed some characteristics of activated cells. After intravenous administration, SBA increased the number of circulating neutrophils and inhibited in a dose-dependent manner the neutrophil migration induced by ip injection of carrageenan into peritoneal cavities. The co-injection of N-acetyl-galactosamine or mannose, but not glucose or fucose, inhibited these effects. The data indicate that soybean lectin is able to induce a local inflammatory reaction but has an anti-inflammatory effect when present in circulating blood

  14. Platelet cold agglutinins and thrombocytopenia: A diagnostic dilemma in the intensive care unit

    TV Bharath Kumar


    Full Text Available We report a case of pseudo-thrombocytopenia due to cold agglutinins against platelets. These cold agglutinins were the cause for diagnostic confusion and resulted in extensive workup and unnecessary therapeutic precautions. A thirty two year old female with Guillain-Barre syndrome was admitted in the ICU and serial work-up showed markedly low levels of platelets. The patient had no symptoms of bleeding and patient was investigated extensively for deciphering the etiology of low platelet count. In-vitro clumping of platelets was suspected and in-vitro studies showed marked clumping of platelets with ethylene-diamine-tetra-acetic acid, citrate and heparinized samples. The manual platelet count was found to be within normal limits. Thrombocytopenia as a result of platelet cold agglutinins is a rare cause of in-vitro low platelet counts. No clinical problems have been reported due to the same.

  15. Reduction of Brucella species and Francisella tularensis cross-reacting agglutinins by dithiothreitol.

    Behan, K A; Klein, G C


    Cross-reaction agglutinin titers to Brucella abortus antigen were found in 42 of 128 tularemia serum specimens, and cross-reaction titers to Francisella tularensis antigen were found in 8 of 34 brucellosis serum specimens. The cross-reaction titers were reduced to 10 or less by dithiothreitol, suggesting that the titers are due to immunoglobulin M antibody.

  16. Apoptosis induction by Maackia amurensis agglutinin in childhood acute lymphoblastic leukemic cells

    Kapoor, Sarika; Marwaha, Ram; Majumdar, Siddhartha;


    Malignant transformation is known to be associated with changes in cell surface carbohydrate-architecture, which can be detected by lectins. In the present study, Maackia amurensis agglutinin (MAA), specific for NeuNAcalpha(2-->3)Gal/GalNAc showed strong binding with lymphoblasts of children havi...

  17. Effect of in vivo γ-irradiation on the binding of wheat germ agglutinin on lymphocyte plasma membranes

    Using quantitative fluorimetry with fluoresceinated wheat germ agglutinin, we have been able to investigate in vivo gamma radiation-induced damage at the outer membrane level of rat splenic lymphocytes, namely damage to the glucosidic moieties of membrane glycoproteins and glycolipids. This paper demonstrates that below an irradiation level of 1 gray (Gy), removal of sialic acid is the major feature leading to new exposed specific binding sites for wheat germ agglutinin, since this lectin is specific for sialic acid and N-acetyl-D-glucosamine. Our studies also suggest that above 1 Gy of irradiation more internal damage occurs, since we observed a striking decrease in wheat germ agglutinin binding sites. (orig.)

  18. The frequency of ABO blood group maternal-fetal incompatibility, maternal iso-agglutinins, and immune agglutinins quantitation in Osogbo, Osun State, South-West of Nigeria

    Oseni Bashiru


    Full Text Available Background : ABO incompatibility in maternal-fetal relationship has been shown to cause hemolytic disease of the newborn (HDNB; a survey which is not yet done in this locality. Aim: Frequency of ABO blood group maternal-fetal incompatibility, maternal iso-agglutinins, and immune agglutinins quantitation was carried out in Osogbo, Osun State, South-West of Nigeria. Settings and Designs : A total of 260 subjects comprising 130 postpartum mothers within the age range of 22-35 years having good obstetrics history and normal delivery, with their 130 neonate babies were used for the study. Materials and Methods : ABO cell and serum groupings were carried out on the subjects using standard antisera and cells with appropriate controls. Direct Coomb′s Test was carried out on neonate red cells. Antibody quantitation by double dilution on the maternal serum using red cells containing corresponding antigen to the antibody was determined. A titer, which is the reciprocal of the highest dilution showing agglutination by Indirect Coombs Test, was determined. Another batch of sera was pretreated with 2-mecarptoethanol before determining the titer. Statistical Analysis: The distribution study results obtained were compared in percentages, whereas the antibodies quantitation was expressed as titers using the mode of the titers for compariso-agglutininsn. Results and Conclusions : Thirty-eight percent (50 mothers were ABO incompatible with their babies, whereas 62% (80 mothers were compatible. The distribution of blood groups in the compatible population showed blood group O (45%; A (30%; B (20%; and AB (5%. Mothers O, A, and B carrying incompatible babies had a frequency of 24% each, whereas mothers AB had 28%. Serologist differences occur in maternal ABO antibodies of corresponding incompatible baby ABO antigens. A high incidence of ABO maternal-fetal incompatibility observed without detection of immune agglutinins is indicative of a rare incidence of HDNB due

  19. Adhesivity and rigidity of erythrocyte membrane in relation to wheat germ agglutinin binding


    Binding of the plant lectin wheat germ agglutinin (WGA) to erythrocyte membranes causes membrane rigidification. One of our objectives has been to directly measure the effects of WGA binding on membrane rigidity and to relate rigidification to the kinetics and levels of WGA binding. Our other objective has been to measure the strength of adhesion and mechanics of cell separation for erythrocytes bound together by WGA. The erythrocyte membrane rigidity was measured on single cells by micropipe...

  20. Nematotoxicity of Marasmius oreades Agglutinin (MOA) Depends on Glycolipid Binding and Cysteine Protease Activity*

    Wohlschlager, Therese; Butschi, Alex; Zurfluh, Katrin; Vonesch, Sibylle C.; auf dem Keller, Ulrich; Gehrig, Peter; Bleuler-Martinez, Silvia; Hengartner, Michael O.; Aebi, Markus; Künzler, Markus


    Fruiting body lectins have been proposed to act as effector proteins in the defense of fungi against parasites and predators. The Marasmius oreades agglutinin (MOA) is a Galα1,3Gal/GalNAc-specific lectin from the fairy ring mushroom that consists of an N-terminal ricin B-type lectin domain and a C-terminal dimerization domain. The latter domain shows structural similarity to catalytically active proteins, suggesting that, in addition to its carbohydrate-binding activity, MOA has an enzymatic ...

  1. O-linked oligosaccharides from salivary agglutinin: Helicobacter pylori binding sialyl-Lewis x and Lewis b are terminating moieties on hyperfucosylated oligo-N-acetyllactosamine

    S. Issa; A.P. Moran; S.N. Ustinov; J.H. Lin; A.J. Ligtenberg; N.G. Karlsson


    Salivary agglutinin plays a vital biological role modulating the protective effect in the oral cavity by interacting with a broad range of oral pathogens. Here, we describe the first characterization of the O-linked oligosaccharides of salivary agglutinin identified by negative ion liquid chromatogr

  2. Enrichment for CFU-C from murine and human bone marrow using soybean agglutinin

    Mouse bone marrow and spleen cells agglutinated by soybean agglutinin (SBA) or peanut agglutinin (PNA) were previously shown to be enriched for spleen colony-forming cells (CFU-S) and sufficiently depleted of graft-versus-host reaction producing cells to allow hematologic reconstitution of lethally irradiated allogeneic recipient mice. A similar enrichment for cells capable of forming colonies in soft agar culture (CFU-C) has now been found in the SBA-agglutinated fraction of mouse bone marrow cells, in contrast to the finding that in human bone marrow the majority of the CFU-C are in the fraction not agglutinated by SBA. Cytofluorometric studies with fluorescein-labeled SBA (FITC-SBA) revealed that the majority of both mouse and human bone marrow cells bind the lectin. Experiments mixing the human marrow fractions separated by SBA reveal that true enrichment for CFU-C is achieved in the unagglutinated fraction, as opposed to a possible depletion of a suppressor cell population. Granulocytic, monocytic, and mixed cell colonies were all enriched in the SBA-unagglutinated cell fraction from human bone marrow

  3. Problems of Cold Agglutinins in Cardiac Surgery: How to Manage Cardiopulmonary Bypass and Myocardial Protection

    Kambiz Alizadeh


    Full Text Available Cold agglutinins are of unique relevance in cardiac surgerybecause of the use of hypothermic cardiopulmonary bypass (CPB. Cold autoimmune diseases are defined by the presence of abnormal circulating proteins (usually IgM or IgA antibodies that agglutinate in response to a decrease in body temperature. These disorders include cryoglobulinemia and cold hemagglutinin disease.Immunoglobulin M autoantibodies to red blood cells, which activateat varying levels of hypothermia, can cause catastrophic hemagglutination,microvascular thrombosis, or hemolysis. Management of anesthesia in these patients includes strict maintenance of normothermia. Patients scheduled for the surgery requiring cardiopulmonary bypass present significant challenges. Use of systemic hypothermia may be contraindicated, and cold cardioplegia solutions may precipitate intracoronary hemagglutination with consequent thrombosis, ischemia, or infarction. Management of CPB andmyocardial protection requires individualized planning. We describea case of MV repair and CABG in a patient with high titercold agglutinins and high thermal amplitude for antibody activation.Normothermic CPB and continuous warm blood cardioplegia weresuccessfully used.


    PANZhi-Xing; WANGYi-Fei


    It has been proved/n our prcvious study that wheat germ agglutinin (WGA) receptor on human sperm membrane is closely related to male fertility and there exists a significant difference in WGA receptors betweea fertile and infertile mcn. In this report, enzyme linked

  5. Salivary agglutinin/DMBT1SAG expression is up-regulated in the presence of salivary gland tumors

    Bikker, F J; van der Wal, J E; Ligtenberg, A J M;


    Salivary agglutinin (SAG) is encoded by the gene Deleted in Malignant Brain Tumors 1 (DMBT1) and represents the salivary variant of DMBT1 (DMBT1(SAG)). While SAG is a bona fide anti-caries factor, DMBT1 was proposed as a candidate tumor-suppressor for brain, digestive tract, and lung cancer. Though...

  6. The Salivary Scavenger and Agglutinin in Early Life: Diverse Roles in Amniotic Fluid and in the Infant Intestine

    Reichhardt, M.P.; Jarva, H.; Been, de M.; Rodriguez, J.M.; Quintana, E.J.; Loimaranta, V.; Vos, de W.M.; Meri, S.


    The salivary scavenger and agglutinin (SALSA), also known as gp340 and dmbt1, is an antimicrobial and inflammation-regulating molecule located at the mucosal surfaces. The present study revealed that SALSA was present in the amniotic fluid (AF) and exceptionally enriched in both meconium and feces o

  7. Expression of Monstera deliciosa agglutinin gene (mda) in tobacco confers resistance to peach-potato aphids.

    Kai, Guoyin; Ji, Qian; Lu, Yang; Qian, Zhongying; Cui, Lijie


    The aphid is one of the most serious pests that causes damage to crops worldwide. Lectins from Araceae plant had been proved useful to control the aphid. Herein, the full-length cDNA of Monstera deliciosa agglutinin (mda) gene was cloned and then introduced into tobacco and the influence of the expression of mda in transgenic tobacco against peach-potato aphids (Myzus persicae) was investigated. Among 92 regenerated plants, 59 positive tobacco lines were obtained. Real-time PCR assays and aphid bioassay test revealed that there is a positive correlation between the expression level of mda and the inhibitory effect on peach-potato aphids. The average anti-pests ability of mda transgenic tobacco was 74%, which was higher than that of other reported lectins from Araceae plant. These results indicated that MDA is one of promising insect resistance proteins selected for the control of peach-potato aphids. PMID:22660606

  8. Application of an Optical Biosensor to Study the Interaction between Porphyrins and Wheat Germ Agglutinin


    An optical biosensor with a stirred cuvette has been used to monitor the interaction of immobilized wheat germ agglutinin (WGA) with two water-soluble cationic porphyrins. The association constants (Ka) of the free base porphyrin and its Zn(Ⅱ) complex form were 2.66 and 27.31×105 L/mol at 20 ℃ respectively. The interactions of the free base porphyrin were further investigated at temperatures between 15 ℃ and 37 ℃. The thermodynamics parameters, changes in free energy, enthalpy and entropy, were -31.23, 22.92, 54.15 kJ/mol respectively. The heat capacity change was -355.53 J·mol-1·K-1. The binding was driven by entropic contribution, and showed strong enthalpy-entropy compensation. It was governed primarily by hydrophobic forces.

  9. Dynamic light scattering study of peanut agglutinin: Size, shape and urea denaturation

    Sagarika Dev; Avadhesha Surolia


    Peanut agglutinin (PNA) is a homotetrameric protein with a unique open quaternary structure. PNA shows non-two state profile in chaotrope induced denaturation. It passes through a monomeric molten globule like state before complete denaturation (Reddy et al 1999). This denaturation profile is associated with the change in hydrodynamic radius of the native protein. Though the molten globule-like state is monomeric in nature it expands in size due to partial denaturation. The size and shape of the native PNA as well as the change in hydrodynamic radius of the protein during denaturation has been studied by dynamic light scattering (DLS). The generation of two species is evident from the profile of hydrodynamic radii. This study also reveals the extent of compactness of the intermediate state.

  10. Cloning and Functional Analysis of the Bifunctional Agglutinin/Trypsin Inhibitor from Helianthus tuberosus L.

    Tuanjie Chang; Hongli Zhai; Songbiao Chen; Guisheng Song; Honglin Xu; Xiaoli Wei; Zhen Zhu


    In order to find new insect resistance genes, four homologous cDNAs, hta-a, hta-b, hta-c and hta-d with lengths of 775, 718, 784 and 752 bp, respectively (GenBank accession numbers AF477031-AF477034), were isolated from a tuber cDNA expression library of Helianthus tuberosus L. Sequence analysis revealed that all four cDNAs contain an open reading frame of 444 bp, coding a polypeptide of 147 amino acid residues, and that the sequences of the cDNAs are very similar to those of the mannose-binding agglutinin genes of the jacalin-related family. In hemagglutination reactions and hapten inhibition assays, affinity-purified HTA (Helianthus tuberosus agglutinin) from induced Escherichia coli BL21(DE3) expressing GST-HTA shows hemagglutination ability and a higher carbohydrate-binding ability for mannose than other tested sugars.Trypsin inhibitory activity was detected in the crude extracts of induced E. coli BL21(DE3)expressing HTA,and was further verified by trypsin inhibitory activity staining on native polyacrylamide gel. The mechanism of interaction between HTA and trypsin was studied by molecular modeling. We found that plenty of hydrogen bonds and electrostatic interactions can be formed between the supposed binding sites of HTA-b and the active site of trypsin, and that a stable HTA/trypsin complex can be formed. The results above imply that HTA might be a bifunctional protein with carbohydrate-binding activity and trypsin inhibitory activity. Moreover,Northern blotting analysis demonstrated that hta is predominantly expressed in tubers of H. tuberosus, very weakly expressed in stems, but not expressed at all in other tissues. Southern blotting analysis indicated that hta is encoded by a multi-gene family. The insect resistance traits have been described in another paper.


    Eliete C. ROMERO


    Full Text Available The persistence of agglutinins detected by MAT has created some problems to the interpretation of the results. The aim of this study was to examine the data of serology from 70 patients with serologically confirmed diagnosis of leptospirosis by during 3-13 months after being affected with leptospires in order to elucidate the interpretation of the persistence of agglutinins detected by MAT. Sixty-one patients sera (87.14% had titers equal or greater than 800. Of these, two individuals maintained titers of 800 thirteen months after the onset. This study showed that only one sample of sera with high titers is not reliable to determine the time at which infection occurred.Persistência de títulos de aglutininas anti-leptospiras em soros humanos diagnosticados pelo teste de aglutinação microscópica A persistência de aglutininas detectadas por MAT tem criado problemas na interpretação dos resultados. O objetivo deste trabalho foi examinar os resultados da sorologia de 70 pacientes com confirmação sorológica de leptospirose durante 3-13 meses após terem sido infectados para se poder elucidar a interpretação da persistência de aglutininas detectadas por MAT. Sessenta e um soros de pacientes (87,14% apresentaram títulos iguais, ou maiores, que 800. Destes, 2 indivíduos mantiveram títulos de 800 treze meses após terem sido infectados. Este estudo mostra que apenas uma amostra de soro, mesmo com alto título de aglutininas, não pode ser considerada para determinar a fase da doença.

  12. Low Prevalence of Brucella Agglutinins in Blood Donors in Central Province of Iran

    Masoomeh Sofian


    Full Text Available Background: Brucellosis is a zoonotic disease of worldwide distribution and has great economic importance. Despite its control in many countries, it remains endemic in Iran. Brucellosis was investigated in many high risk occupational groups; however, few studies on the prevalence of brucellosis among blood donors are available. To determine the seroprevalence of brucellosis antibodies in blood donors, a serological study was carried out in central province of Iran.Materials and Methods: A total of 897 healthy blood donors with mean age 37.23 ± 10.9 years were enrolled in the study. Laboratory tests including Standard Tube Agglutination Test (STA and 2-mercaptoethanol (2ME agglutination were checked in all samples. STA dilution ≥ 1:80, and in the presence of 2-mercaptoethanol (2ME agglutination ≥ 20 was considered positive.Results: Out of 897 cases, 11.9% were inhabitants of rural areas. 41.5% had history of consumption of unpasteurized dairy products and 9.3% had history of contact with domestic animals. A very low level of Brucella agglutinins was present in 3(0.33% of the samples and only one sample (0.11% was found to be truly positive for Brucella agglutinins. 2ME was negative in all samples. None of these 4 subjects showed signs and symptoms of brucellosis in 6 months follow-up.Conclusion: On the basis of our data, brucellosis has no epidemiological and clinical importance in our blood donors; therefore, it is not recommended to perform screening tests such as, STA and 2ME to identify brucellosis antibodies in the sera of blood donors.

  13. Agglutinins and cardiac surgery: a web based survey of cardiac anaesthetic practice; questions raised and possible solutions

    Shah, S.; Gilliland, H; Benson, G


    Introduction Cardiac surgery involves cardiopulmonary bypass during which the core temperature is generally lowered to hypothermic levels. Patients presenting for cardiac surgery are sometimes reported to have cold or warm autoantibodies at the time of blood screening. It is known that cold agglutinins may cause potentially life-threatening haemolysis, intracoronary haemagglutination leading to inadequate cardioplegia distribution, thrombosis, embolism, ischaemia or infarction. The risk (if a...

  14. Discovering the Secrets of the Candida albicans Agglutinin-Like Sequence (ALS) Gene Family—a Sticky Pursuit

    HOYER, LOIS L.; GREEN, CLAYTON B.; Oh, Soon-Hwan; Zhao, Xiaomin


    The Agglutinin-Like Sequence (ALS) family of Candida albicans includes eight genes that encode large cell-surface glycoproteins. The high degree of sequence relatedness between the ALS genes and the tremendous allelic variability often present in the same C. albicans strain complicated definition and characterization of the gene family. The main hypothesis driving ALS family research is that the genes encode adhesins, primarily involved in host-pathogen interactions. Although adhesive functio...

  15. Purification and partial characterization of an agglutinin from Octopus maya serum.

    Alpuche, Juan; Pereyra, Ali; Mendoza-Hernández, Guillermo; Agundis, Concepción; Rosas, Carlos; Zenteno, Edgar


    A 66-kDa lectin (OmA) was purified from the serum of the Yucatan peninsula endemic octopus (Octopus maya) by a single step affinity chromatography on glutaraldehyde-fixed stroma from rat erythrocytes. OmA corresponds to 0.8% of the total circulating protein in the hemolymph; it is composed of three equal subunits of 22kDa each, and 7.4% of linked carbohydrates. The amino acids' composition indicated that agglutinin contained mainly aspartic and glutamic acids, and cysteine and methionine were identified in minor proportion. OmA agglutinates mainly rat, guinea pig, and rabbit erythrocytes, and this activity is partially inhibited by galactosamine, melobiose, galacturonic acid, mannose, and methyl alpha and beta galactosides. Hemagglutinating activity is not dependent on divalent cations, such as Ca(2+), Mg(2+), or Mn(2+). The OmA subunits showed no identity for any lectin in databases but partial identity with the type A hemocyanin from Octopus dolfleini hemolymph; the main similarities are related to tyrosinase domains and copper A and B sites that conform to the oxygen-binding site of hemocyanin. PMID:20105460

  16. Transneuronally transported wheat germ agglutinin labels glia as well as neurons in the rat visual system

    Following intraocular injection in adult rats of 125I-labeled wheat germ agglutinin (I-WGA), the ultrastructural distribution of label in the superior colliculus and lateral geniculate was examined by electron microscope autoradiography. Three days after injection, 5.4% of the label in the lateral geniculate was associated with neuronal perikarya, and 3.6% was associated with glial perikarya. The corresponding figures for the superior colliculus were 5.1% and 0.8%. When the data were expressed as number of grains per micron 2 cytoplasm, there was no statistically significant difference between the grain density over neuronal or glial cytoplasm in either the lateral geniculate or the superior colliculus. A statistical analysis of the distance between the silver grains and the cell membranes showed that in both neurons and glia, the observed labeling was the product of internalized I-WGA and not the result of scatter from the neuropil or from label bound to the surface of the cells. These results indicate that much of the WGA released from axons and axon terminals is not confined to a specific ''transsynaptic'' pathway, but produces a generalized labeling of nearby cells, much like a microinjection of WGA into the region

  17. The prevalence of anti-leptospiral agglutinins in sera of wildlife in southeastern Australia.

    Milner, A R; Wilks, C R; Spratt, D M; Presidente, P J


    Anti-leptospiral agglutinins were found in the serum from 18 (7 species) of 419 (25 species) animals sampled from various areas of southeastern Australia. Positive serologic reactions were observed in 5 of 25 (20%) brush-tailed possum (Trichosurus vulpecula), 1 of 26 (3.8%) tammar wallaby (Macropus eugenii), 2 of 12 (16.7%) swamp wallaby (Wallabia bicolor), 1 of 3 (33.3%) koala (Phascolarctos cinereus), 3 of 41 (7.3%) common wombat (Vombatus ursinus), 2 of 100 (2%) bush rat (Rattus fuscipes) and 4 of 12 (25%) rusa deer (Cervus timorensis). The majority (55.5%) of serologic reactions were to serovar hardjo. No serologic reactions were observed in samples from echidna (Tachyglossus aculeatus), brown antechinus (Antechinus stuartii), swainson's antechinus (Antechinus swaisonsii), long-nosed bandicoot (Perameles nasuta), brown bandicoot(Isoodon obesulus), common ringtail (Pseudocheirus peregrinus), greater glider (Schoinobates volans), eastern grey kangaroo (Macropus giganteus), red-necked wallaby (Macropus rufogriseus), rabbit (Oryctolagus cuniculus), water rat (Hydromys chrysogaster), black rat (Rattus rattus), eastern swamp rat (Rattus lutreolus), broad-toothed rat (Mastacomys fuscus), fox (Vulpes vulpes), sambar deer (Cervus unicolor), hog deer (Axis porcinus) and fallow deer (Dama dama). PMID:7241704

  18. Evolution of the rapidly mutating human salivary agglutinin gene (DMBT1) and population subsistence strategy.

    Polley, Shamik; Louzada, Sandra; Forni, Diego; Sironi, Manuela; Balaskas, Theodosius; Hains, David S; Yang, Fengtang; Hollox, Edward J


    The dietary change resulting from the domestication of plant and animal species and development of agriculture at different locations across the world was one of the most significant changes in human evolution. An increase in dietary carbohydrates caused an increase in dental caries following the development of agriculture, mediated by the cariogenic oral bacterium Streptococcus mutans. Salivary agglutinin [SAG, encoded by the deleted in malignant brain tumors 1 (DMBT1) gene] is an innate immune receptor glycoprotein that binds a variety of bacteria and viruses, and mediates attachment of S. mutans to hydroxyapatite on the surface of the tooth. In this study we show that multiallelic copy number variation (CNV) within DMBT1 is extensive across all populations and is predicted to result in between 7-20 scavenger-receptor cysteine-rich (SRCR) domains within each SAG molecule. Direct observation of de novo mutation in multigeneration families suggests these CNVs have a very high mutation rate for a protein-coding locus, with a mutation rate of up to 5% per gamete. Given that the SRCR domains bind S. mutans and hydroxyapatite in the tooth, we investigated the association of sequence diversity at the SAG-binding gene of S. mutans, and DMBT1 CNV. Furthermore, we show that DMBT1 CNV is also associated with a history of agriculture across global populations, suggesting that dietary change as a result of agriculture has shaped the pattern of CNV at DMBT1, and that the DMBT1-S. mutans interaction is a promising model of host-pathogen-culture coevolution in humans. PMID:25848046

  19. A novel autotransporter of uropathogenic Proteus mirabilis is both a cytotoxin and an agglutinin.

    Alamuri, Praveen; Mobley, Harry L T


    One of the six predicted Proteus mirabilis autotransporters (ATs), ORF c2341, is predicted to contain a serine protease motif and was earlier identified as an immunogenic outer membrane protein in P. mirabilis. The 3.2 kb gene encodes a 117 kDa protein with a 58-amino-acid-long signal peptide, a 75-kDa-long N-terminal passenger domain and a 30-kDa-long C-terminal translocator. Affinity-purified 110 kDa AT exhibited chymotrypsin-like activity and hydrolysed N-Suc-Ala-Ala-Pro-Phe-pNa and N-Suc-Ala-Ala-Pro-Leu-pNa with a K(M) of 22 muM and 31 muM, respectively, under optimal pH of 8.5-9.0 in a Ca(2+)-dependent manner. Activity was inhibited by subtilase-specific inhibitors leupeptin and chymostatin. Both the cell-associated and purified form elicited cytopathic effects on cultured kidney and bladder epithelial cells. Substrate hydrolysis as well as cytotoxicity was associated with the passenger domain and was compromised upon mutation of any of the catalytic residues (Ser366, His147 and Asp533). At alkaline pH and optimal cell density, the AT also promoted autoaggregation of P. mirabilis and this function was independent of its protease activity. Cytotoxicity, autoaggregation and virulence were significantly reduced in an isogenic pta mutant of P. mirabilis. Proteus toxic agglutinin (Pta) represents a novel autotransported cytotoxin with no bacterial homologues that works optimally in the alkalinized urinary tract, a characteristic of urease-mediated urea hydrolysis during P. mirabilis infection. PMID:18430084

  20. Effect of wheat germ agglutinin on the viscoelastic properties of erythrocyte membrane.

    Smith, L; Hochmuth, R M


    The elasticity and viscosity of the human erythrocyte membrane were measured as a function of the concentration of wheat germ agglutinin (WGA) in a suspending solution containing 1 mg/ml albumin, approximately 5 X 10(5) cells/ml and between 0.0 and 0.2 microgram/ml WGA. Membrane elasticity was characterized by the elastic shear modulus, which provided a measure of the resistance of the membrane to constant-area elastic deformations that occurred in the membrane plane. The elastic shear modulus was determined by aspirating a portion of the membrane into a micropipette and measuring the extension of the membrane into the pipette as a function of the suction pressure. The results indicated no significant change in shear modulus for concentrations of WGA between 0.0 and 0.2 microgram/ml. Membrane viscosity was characterized by the coefficient of surface viscosity, which, in effect, was a measure of the membrane's resistance to rates of deformation. This coefficient was determined from the time required for an erythrocyte to recover its undeformed shape after it had been elongated by the application of an equal and opposite force applied at diametrically opposite points on the erythrocyte rim. The value for the coefficient of surface viscosity was found to increase by a factor of almost three when the WGA concentration was increased from 0.0 to 0.2 microgram/ml. These results indicated that, in the presence of albumin, WGA can increase membrane dissipation (viscosity) without altering the structural rigidity (elasticity) of the membrane. PMID:6896878

  1. Vicia villosa agglutinin separates freshly isolated Peyer's Patch T cells into interleukin 5- or interleukin 2-producing subsets


    Murine L3T4 T cells freshly isolated from Peyer's Patch were fractionated based on differential adherence to Vicia villosa agglutinin (VVA). VVA adherent cells secreted IL-5, but not IL-2, after stimulation with Con A and IL-1. In striking contrast, VVA nonadherent PP L3T4 T cells secreted IL-2, but not IL-5, under the same conditions. In addition, supernatants from VVA adherent, but not from VVA nonadherent cells cultures, enhanced IgA secretion by LPS-stimulated splenic B cells to the same ...

  2. Evaluation of the Role of Candida albicans Agglutinin-Like Sequence (Als) Proteins in Human Oral Epithelial Cell Interactions

    Murciano, Celia; Moyes, David L.; Runglall, Manohursingh; Tobouti, Priscila; Islam, Ayesha; HOYER, LOIS L.; Naglik, Julian R.


    The fungus C. albicans uses adhesins to interact with human epithelial surfaces in the processes of colonization and pathogenesis. The C. albicans ALS (agglutinin-like sequence) gene family encodes eight large cell-surface glycoproteins (Als1-Als7 and Als9) that have adhesive function. This study utilized C. albicans Δals mutant strains to investigate the role of the Als family in oral epithelial cell adhesion and damage, cytokine induction and activation of a MAPK-based (MKP1/c-Fos) signalin...

  3. Identification of the bacteria-binding peptide domain on salivary agglutinin (gp-340/DMBT1), a member of the scavenger receptor cysteine-rich superfamily

    Bikker, Floris J; Ligtenberg, Antoon J M; Nazmi, Kamran;


    Salivary agglutinin is encoded by DMBT1 and identical to gp-340, a member of the scavenger receptor cysteine-rich (SRCR) superfamily. Salivary agglutinin/DMBT1 is known for its Streptococcus mutans agglutinating properties. This 300-400 kDa glycoprotein is composed of conserved peptide motifs: 14...... containing exclusively SRCR and SID domains that binds to S. mutans. To define more closely the S. mutans-binding domain, consensus-based peptides of the SRCR domains and SIDs were designed and synthesized. Only one of the SRCR peptides, designated SRCRP2, and none of the SID peptides bound to S. mutans....... Strikingly, this peptide was also able to induce agglutination of S. mutans and a number of other bacteria. The repeated presence of this peptide in the native molecule endows agglutinin/DMBT1 with a general bacterial binding feature with a multivalent character. Moreover, our studies demonstrate for the...

  4. Studies on glycoproteins produced by wild type and wheat germ agglutinin-resistant B16 mouse melanoma cells

    Two variants of B16 mouse melanoma cells have been selected in serum-free medium for their resistance to toxic levels of wheat germ agglutinin isolation 1 (WGA). Chromosome analysis and characteristic melanin production showed that the variants are derived from the parent mouse melanoma cell lines. However, the two variants were less tumorigenic in mice compared to the parent B16 mouse melanoma cells. The variants showed a marked decrease in cell agglutination with WGA. Cell agglutination with recin and peanut lectin was not different between the three cell lines, but the two variants showed a slight increase in agglutination with concanavalin A. The binding of 125I-labeled wheat germ agglutinin to the two variant cells was reduced compared to that of the parent cell. Glycoproteins secreted or shed by the three lines were isolated after growth in serum-free medium in the presence of [3He]glucosamine and bovine serum albumin (1%). These metabolically labeled products were fractionated on the basis of their interaction with WGA-Sepharose (2 mg/ml). The WGA-Sepharose affinity chromatographic data suggested a decrease in WGA-binding glycoprotein(s) secreted to the medium by the two variants. The WGA-bound glycoproteins from the two variants upon SDS-PAGE revealed three bands of approximate molecular weights, 92,000, 56,000, and 42,000, none of which were present in the parent cell line (50,000 molecular weight)

  5. Prevalence and spatial analysis of antileptospiral agglutinins in dairy cattle - Microregion of Sete Lagoas, Minas Gerais, 2009/2010

    R.R. Nicolino


    Full Text Available The aim of this study was to evaluate the seroprevalence of leptospirosis in the dairy herds from Minas Gerais, Brazil, during the years 2009 and 2010. A total of 2,915 serum samples were collected from the lactating cows of 151 properties in eleven municipalities located in the Sete Lagoas region. The Microscopic Agglutination Test was used to detect antileptospiral agglutinins. An individual animal prevalence of 20.7% (95% CI = 17.1% - 24.3% and a herd prevalence of 80.8% (95% CI = 73.8% = 87.7% were determined. The most prevalent serovars were hardjoprajitno at 19.4%; hardjoprajitno strain Norma at 17.4%; and hardjo-bovis at 17.4%. These results show the significance of the hardjo serovar in bovine leptospirosis cases in Minas Gerais.

  6. Transgenic tetraploid Isatis indigotica expressing Bt Cry1Ac and Pinellia ternata agglutinin showed enhanced resistance to moths and aphids.

    Xiao, Ying; Wang, Kai; Ding, Ruxian; Zhang, Hanming; Di, Peng; Chen, Junfeng; Zhang, Lei; Chen, Wansheng


    Co-expression of multiple genes encoding different kinds of insect resistant proteins has been developed to confer a broader spectrum of pest control. Tetraploid Isatis indigotica Fort was transformed with a plasmid, p3300BP, containing Bacillus thuringiensis Cry1Ac gene (Bt) and Pinellia ternata agglutinin gene (Pta) and the selectable marker herbicide resistance gene (Bar) driven by the CaMV35S promoter via Agrobacterium tumefaciens-mediated transformation. The integration and expression of introduced genes in regenerated transgenic plants were confirmed by PCR and Western blot assays. Insect bioassay test demonstrated transgenic lines had significant inhibition to diamondback moths (Plutella xylostella L.) and peach potato aphids (Myzus persicae Sulzer) simultaneously. Our study reported here would be a great motivation for field culture of tetraploid I. indigotica, also providing an efficient molecular breeding strategy to provide insect tolerant plants. PMID:21559837

  7. An Unusual Member of the Papain Superfamily: Mapping the Catalytic Cleft of the Marasmius oreades agglutinin (MOA) with a Caspase Inhibitor

    Gabriele Cordara; André van Eerde; Grahn, Elin M.; Harry C Winter; Goldstein, Irwin J.; Ute Krengel


    Papain-like cysteine proteases (PLCPs) constitute the largest group of thiol-based protein degrading enzymes and are characterized by a highly conserved fold. They are found in bacteria, viruses, plants and animals and involved in a number of physiological and pathological processes, parasitic infections and host defense, making them interesting targets for drug design. The Marasmius oreades agglutinin (MOA) is a blood group B-specific fungal chimerolectin with calcium-dependent proteolytic a...

  8. Measurement of the Fucosylated Sugar Chains of Serum α_1-Antitrypsin : The Relationship between Reactivity with Lens Culinaris Agglutinin and HPLC Analysis of the Pyridylaminated Sugar Chains

    Saitoh, Atsushi; Aoyagi, Yutaka; Mori, Shigeki; Suda, Takeshi; Suzuki, Yasufumi; Sekine, Chuichi; Asakura, Hitoshi


    Twelve samples of α_1-anlitrypsin (AAT) were purified from six patients with hepatocellular carcinoma, one with pancreatic cancer and five healthy individuals, and subjected to the measurement of the fucosylated sugar chain. We compared different results from crossed immunoaffinoelectrophoresis (CIAE) with Lens culinaris agglutinin (LCA) and by the carbohydrate analysis of high-performance liquid chromatography (HPLC) with the pylidylaminated (PA・) oligosaccharides obtained from each AAT. A s...

  9. Allergenicity assessment of Allium sativum leaf agglutinin, a potential candidate protein for developing sap sucking insect resistant food crops.

    Hossain Ali Mondal

    Full Text Available BACKGROUND: Mannose-binding Allium sativum leaf agglutinin (ASAL is highly antinutritional and toxic to various phloem-feeding hemipteran insects. ASAL has been expressed in a number of agriculturally important crops to develop resistance against those insects. Awareness of the safety aspect of ASAL is absolutely essential for developing ASAL transgenic plants. METHODOLOGY/PRINCIPAL FINDINGS: Following the guidelines framed by the Food and Agriculture Organization/World Health Organization, the source of the gene, its sequence homology with potent allergens, clinical tests on mammalian systems, and the pepsin resistance and thermostability of the protein were considered to address the issue. No significant homology to the ASAL sequence was detected when compared to known allergenic proteins. The ELISA of blood sera collected from known allergy patients also failed to show significant evidence of cross-reactivity. In vitro and in vivo assays both indicated the digestibility of ASAL in the presence of pepsin in a minimum time period. CONCLUSIONS/SIGNIFICANCE: With these experiments, we concluded that ASAL does not possess any apparent features of an allergen. This is the first report regarding the monitoring of the allergenicity of any mannose-binding monocot lectin having insecticidal efficacy against hemipteran insects.

  10. Abrus agglutinin is a potent anti-proliferative and anti-angiogenic agent in human breast cancer.

    Bhutia, Sujit K; Behera, Birendra; Nandini Das, Durgesh; Mukhopadhyay, Subhadip; Sinha, Niharika; Panda, Prashanta Kumar; Naik, Prajna Paramita; Patra, Samir K; Mandal, Mahitosh; Sarkar, Siddik; Menezes, Mitchell E; Talukdar, Sarmistha; Maiti, Tapas K; Das, Swadesh K; Sarkar, Devanand; Fisher, Paul B


    Abrus agglutinin (AGG), a plant lectin isolated from the seeds of Abrus precatorius, has documented antitumor and immunostimulatory effects in murine models. To examine possible antitumor activity against breast cancer, we established human breast tumor xenografts in athymic nude mice and intraperitoneally administered AGG. AGG inhibited tumor growth and angiogenesis as confirmed by monitoring the expression of Ki-67 and CD-31, respectively. In addition, TUNEL positive cells increased in breast tumors treated with AGG suggesting that AGG mediates anti-tumorigenic activity through induction of apoptosis and inhibition of angiogenesis. On a molecular level, AGG caused extrinsic apoptosis through ROS generation that was AKT-dependent in breast cancer cells, without affecting primary mammary epithelial cells, suggesting potential cancer specificity of this natural compound. In addition, using HUVECs, AGG inhibited expression of the pro-angiogenic factor IGFBP-2 in an AKT-dependent manner, reducing angiogenic phenotypes both in vitro and in vivo. Overall, the present results establish that AGG promotes both apoptosis and anti-angiogenic activities in human breast tumor cells, which might be exploited for treatment of breast and other cancers. PMID:26914517

  11. Synthesis of tetravalent LacNAc-glycoclusters as high-affinity cross-linker against Erythrina cristagalli agglutinin.

    Ogata, Makoto; Chuma, Yasushi; Yasumoto, Yoshinori; Onoda, Takashi; Umemura, Myco; Usui, Taichi; Park, Enoch Y


    Four kinds of tetravalent double-headed glycoclusters [(LacNAc)4-DHGs] were designed with linkers of varying lengths consisting of alkanedioic carboxyamido groups (C6, C12, C18 and C24) between two bi-antennary LacNAc-glycosides. These glycoclusters served as high-affinity cross-linking ligands for the LacNAc-binding lectin Erythrina cristagalli agglutinin (ECA). The binding activity and cross-linking between each ligand and ECA were characterized by a hemagglutination inhibition (HI) assay, isothermal titration calorimetry (ITC), a quantitative precipitation assay and dynamic light scattering (DLS). For the precipitation assay and DLS measurement, the synthesized (LacNAc)4-DHGs were found to be capable of binding and precipitating the ECA as multivalent ligands. ITC analysis indicated the binding of (LacNAc)4-DHGs was driven by a favorable enthalpy change. Furthermore, the entropy penalty from binding (LacNAc)4-DHGs clearly decreased in a spacer length-dependent manner. The binding affinities of flexible (LacNAc)4-DHGs (C18 and C24) with long spacers were found to be more favorable than those of the clusters having short spacers (C6 and C12). These results were supported by molecular dynamics simulations with explicit water molecules for the tetravalent glycoclusters with ECA. We concluded that the subtle modification in the epitope-presenting scaffolds exerts the significant effect in the recognition efficiency involved in the LacNAc moieties by ECA. PMID:26672510

  12. Effects of wheat germ agglutinin on human gastrointestinal epithelium: Insights from an experimental model of immune/epithelial cell interaction

    Wheat germ agglutinin (WGA) is a plant protein that binds specifically to sugars expressed, among many others, by human gastrointestinal epithelial and immune cells. WGA is a toxic compound and an anti-nutritional factor, but recent works have shown that it may have potential as an anti-tumor drug and as a carrier for oral drugs. To quantitate the toxicity threshold for WGA on normal epithelial cells we previously investigated the effects of the lectin on differentiated Caco2 cells, and showed that in the micromolar range of concentrations WGA could alter the integrity of the epithelium layer and increase its permeability to both mannitol and dextran. WGA was shown to be uptaken by Caco2 cells and only ∼ 0.1% molecules were observed to cross the epithelium layer by transcytosis. Here we show that at nanomolar concentrations WGA is unexpectedly bioactive on immune cells. The supernatants of WGA-stimulated peripheral blood mononuclear cells (PBMC) can alter the integrity of the epithelium layer when administered to the basolateral side of differentiated Caco2 cells and the effects can be partially inhibited by monoclonal antibodies against IL1, IL6 and IL8. At nanomolar concentrations WGA stimulates the synthesis of pro-inflammatory cytokines and thus the biological activity of WGA should be reconsidered by taking into account the effects of WGA on the immune system at the gastrointestinal interface. These results shed new light onto the molecular mechanisms underlying the onset of gastrointestinal disorders observed in vivo upon dietary intake of wheat-based foods.

  13. Development of transgenic cotton lines expressing Allium sativum agglutinin (ASAL for enhanced resistance against major sap-sucking pests.

    Chakravarthy S K Vajhala

    Full Text Available Mannose-specific Allium sativum leaf agglutinin encoding gene (ASAL and herbicide tolerance gene (BAR were introduced into an elite cotton inbred line (NC-601 employing Agrobacterium-mediated genetic transformation. Cotton transformants were produced from the phosphinothricin (PPT-resistant shoots obtained after co-cultivation of mature embryos with the Agrobacterium strain EHA105 harbouring recombinant binary vector pCAMBIA3300-ASAL-BAR. PCR and Southern blot analysis confirmed the presence and stable integration of ASAL and BAR genes in various transformants of cotton. Basta leaf-dip assay, northern blot, western blot and ELISA analyses disclosed variable expression of BAR and ASAL transgenes in different transformants. Transgenes, ASAL and BAR, were stably inherited and showed co-segregation in T1 generation in a Mendelian fashion for both PPT tolerance and insect resistance. In planta insect bioassays on T2 and T3 homozygous ASAL-transgenic lines revealed potent entomotoxic effects of ASAL on jassid and whitefly insects, as evidenced by significant decreases in the survival, development and fecundity of the insects when compared to the untransformed controls. Furthermore, the transgenic cotton lines conferred higher levels of resistance (1-2 score with minimal plant damage against these major sucking pests when bioassays were carried out employing standard screening techniques. The developed transgenics could serve as a potential genetic resource in recombination breeding aimed at improving the pest resistance of cotton. This study represents the first report of its kind dealing with the development of transgenic cotton resistant to two major sap-sucking insects.

  14. An atypical IgM class platelet cold agglutinin induces GPVI-dependent aggregation of human platelets.

    Sánchez Guiu, I M; Martínez-Martinez, I; Martínez, C; Navarro-Fernandez, J; García-Candel, F; Ferrer-Marín, F; Vicente, V; Watson, S P; Andrews, R K; Gardiner, E E; Lozano, M L; Rivera, J


    Platelet cold agglutinins (PCA) cause pseudothrombocytopenia, spurious thrombocytopenia due to ex vivo platelet clumping, complicating clinical diagnosis, but mechanisms and consequences of PCA are not well defined. Here, we characterised an atypical immunoglobulin (Ig)M PCA in a 37-year-old woman with lifelong bleeding and chronic moderate thrombocytopenia, that induces activation and aggregation of autologous or allogeneic platelets via interaction with platelet glycoprotein (GP)VI. Patient temperature-dependent pseudothrombocytopenia was EDTA-independent, but was prevented by integrin αIIbβ3 blockade. Unstimulated patient platelets revealed elevated levels of bound IgM, increased expression of activation markers (P-selectin and CD63), low GPVI levels and abnormally high thromboxane (TX)A2 production. Patient serum induced temperature- and αIIbβ3-dependent decrease of platelet count in allogeneic donor citrated platelet-rich plasma (PRP), but not in PRP from Glanzmann's thrombasthenia or afibrinogenaemia patients. In allogeneic platelets, patient plasma induced shape change, P-selectin and CD63 expression, (14)C-serotonin release, and TXA2 production. Activation was not inhibited by aspirin, cangrelor or blocking anti-Fc receptor (FcγRIIA) antibody, but was abrogated by inhibitors of Src and Syk, and by a soluble GPVI-Fc fusion protein. GPVI-deficient platelets were not activated by patient plasma. These data provide the first evidence for an IgM PCA causing platelet activation/aggregation via GPVI. The PCA activity persisted over a five-year follow-up period, supporting a causative role in patient chronic thrombocytopenia and bleeding. PMID:25994029

  15. Wheat germ agglutinin-conjugated PLGA nanoparticles for enhanced intracellular delivery of paclitaxel to colon cancer cells.

    Wang, Chunxia; Ho, Paul C; Lim, Lee Yong


    The purpose of this study was to investigate the potentiation of the anticancer activity and enhanced cellular retention of paclitaxel-loaded PLGA nanoparticles after surface conjugation with wheat germ agglutinin (WGA) against colon cancer cells. Glycosylation patterns of representative colon cancer cells confirmed the higher expression levels of WGA-binding glycoproteins in the Caco-2 and HT-29 cells, than in the CCD-18Co cells. Cellular uptake and in vitro cytotoxicity of WNP (final formulation) against colon cell lines was evaluated alongside control formulations. Confocal microscopy and quantitative analysis of intracellular paclitaxel were used to monitor the endocytosis and retention of nanoparticles inside the cells. WNP showed enhanced anti-proliferative activity against Caco-2 and HT-29 cells compared to corresponding nanoparticles without WGA conjugation (PNP). The greater efficacy of WNP was associated with higher cellular uptake and sustained intracellular retention of paclitaxel, which in turn was attributed to the over-expression of N-acetyl-D-glucosamine-containing glycoprotein on the colon cell membrane. WNP also demonstrated increased intracellular retention in the Caco-2 (30% of uptake) and HT-29 (40% of uptake) cells, following post-uptake incubation with fresh medium, compared to the unconjugated PNP nanoparticles (18% in Caco-2) and (27% in HT-29), respectively. Cellular trafficking study of WNP showed endocytosed WNP could successful escape from the endo-lysosome compartment and release into the cytosol with increasing incubation time. It may be concluded that WNP has the potential to be applied as a targeted delivery platform for paclitaxel in the treatment of colon cancer. PMID:20804835

  16. Labelling of neurons in the rat superior cervical ganglion after injection of wheat-germ agglutinin-horseradish peroxidase into the contralateral ganglion: evidence of transneuronal labelling.

    ATASEVER, A.; Palaoğlu, S; Erbengi, A; Celik, H H


    Recent studies have shown that injection of the tracer wheat-germ agglutinin-horseradish peroxidase (WGA-HRP) into the superior cervical ganglion (SCG) of one side results in labelling of neurons in the contralateral SCG and the stellate ganglion. This study was designed to verify whether or not bilateral projections from the superior cervical ganglion to the midline structures, particularly to the pineal gland, play a role in the transport of WGA-HRP to the contralateral SCG. One group of ra...

  17. Differences in the labeling of axons of passage by wheat germ agglutinin after uptake by cut peripheral nerve versus injections within the central nervous system

    Injections of a radiolabeled derivative of wheat germ agglutinin (WGA) in the subcortical white matter of the cerebral cortex in mice do not give rise to autoradiographic labeling of axon systems coursing through this fiber mass. Exposing the cut-end of mouse tibial nerve to WGA does, however, produce labeling within dorsal root ganglia and the spinal cord. These findings are discussed with consideration for dissimilarity in mode of central versus peripheral administration of the tracer, as well as in the light of potential relative differences in the uptake and transport of WGA and HRP. (Auth.)

  18. Early Serologic Diagnosis of Mycoplasma pneumoniae Pneumonia: An Observational Study on Changes in Titers of Specific-IgM Antibodies and Cold Agglutinins

    Lee, Sung-Churl; Youn, You-Sook; Rhim, Jung-Woo; Kang, Jin-Han; Lee, Kyung-Yil


    Abstract There have been some limitations on early diagnosis of Mycoplasma pneumoniae (MP) infection because of no immunoglobulin M (IgM) responses and variable detection rates of polymerase chain reaction in the early stage of the disease. We wanted to discuss regarding early diagnostic method using short-term paired titration of MP-specific IgM and cold agglutinins (CAs) in the early stage of MP pneumonia. The participants of this study were 418 children with MP pneumonia during 2 recent ep...

  19. Expression of peanut agglutinin-binding mucin-type glycoprotein in human esophageal squamous cell carcinoma as a marker

    Balakrishnan Ramathilakam


    Full Text Available Abstract Background The TF (Thomson – Friedenreich blood group antigen behaves as an onco-foetal carcinoma-associated antigen, showing increased expression in malignancies and its detection and quantification can be used in serologic diagnosis mainly in adenocarcinomas. This study was undertaken to analyze the sera and tissue level detectable mucin-type glycoprotein (TF-antigen by Peanut agglutinin (PNA and its diagnostic index in serum as well tissues of human esophageal squamous cell carcinoma as marker. Results We examined 100 patients for serological analysis by Enzyme Linked Lectin Assay (ELISA and demonstrated a sensitivity of 87.5%, specificity of 90% and a positive predictive value of 95%. The immuno-histochemical localization of TF antigen by Fluorescence Antigen Technique (FAT in 25 specimens of normal esophageal squamous epithelium specimens and 92 specimens with different grades of, allowed a quicker and more precise identification of its increased expression and this did not correlate with gender and tumor size. There was a positive correlation between membrane bound TF antigen expression with different histological progression, from well differentiated to poorly differentiated, determined by PNA binding. Specimens showed morphological changes and a pronounced increase in PNA binding in Golgi apparatus, secretory granules of the cytosol of well differentiated and an increased cell membrane labeling in moderately and poorly differentiated, when compared with ESCC and normal tissues. Conclusion The authors propose that the expression of TF-antigen in human may play an important role during tumorigenesis establishing it as a chemically well-defined carcinoma-associated antigen. Identification of the circulating TF-antigen as a reactive form and as a cryptic form in the healthy individuals, using PNA-ELLA and Immunohistochemical analysis of TF antigen by FAT is positively correlated with the different histological grades as a simple

  20. Transgenic rice expressing Allium sativum leaf agglutinin (ASAL exhibits high-level resistance against major sap-sucking pests

    Vudem Dasavantha


    Full Text Available Abstract Background Rice (Oryza sativa productivity is adversely impacted by numerous biotic and abiotic factors. An approximate 52% of the global production of rice is lost annually owing to the damage caused by biotic factors, of which ~21% is attributed to the attack of insect pests. In this paper we report the isolation, cloning and characterization of Allium sativum leaf agglutinin (asal gene, and its expression in elite indica rice cultivars using Agrobacterium-mediated genetic transformation method. The stable transgenic lines, expressing ASAL, showed explicit resistance against major sap-sucking pests. Results Allium sativum leaf lectin gene (asal, coding for mannose binding homodimeric protein (ASAL from garlic plants, has been isolated and introduced into elite indica rice cultivars susceptible to sap-sucking insects, viz., brown planthopper (BPH, green leafhopper (GLH and whitebacked planthopper (WBPH. Embryogenic calli of rice were co-cultivated with Agrobacterium harbouring pSB111 super-binary vector comprising garlic lectin gene asal along with the herbicide resistance gene bar, both under the control of CaMV35S promoter. PCR and Southern blot analyses confirmed stable integration of transgenes into the genomes of rice plants. Northern and western blot analyses revealed expression of ASAL in different transgenic rice lines. In primary transformants, the level of ASAL protein, as estimated by enzyme-linked immunosorbent assay, varied between 0.74% and 1.45% of the total soluble proteins. In planta insect bioassays on transgenic rice lines revealed potent entomotoxic effects of ASAL on BPH, GLH and WBPH insects, as evidenced by significant decreases in the survival, development and fecundity of the insects. Conclusion In planta insect bioassays were carried out on asal transgenic rice lines employing standard screening techniques followed in conventional breeding for selection of insect resistant plants. The ASAL expressing rice

  1. Distribution features of lymphocytes with peanut agglutinin positive receptors in gums epithelium of rats in norm and after intrauterine antigenic action

    Burega Yu.


    Full Text Available Background. According to the conception “Lymphocyte is the main factor of morphogenesis” changes in lymphocyte receptor repertory, induced by antigenic action in the fetal period of development, influence on organs and tissues development after birth. Functional activity of immunological immature PNA+ lymphocytes inducing the change in functioning, imbalance in formation cells of microenvironment, synthesis of intracellular substance and the fibers of extracellular matrix leads to violation of morphological and functional condition of organs. Objective. Determine the features of distribution of lymphocytes with receptors to peanut agglutinin in gingival epithelium of rats in norm and after intrauterine antigenic action. Methods. The object of the research was: 224 jaws of 112 white laboratory rats. The rats divided into three groups. First group – intact rats. Second group –rats, which were introduced 0,05 ml solution of antigen in the amniotic fluid on the 18th day of pregnancy by the method of N. Voloshyn, the third group – control, the animals were introduced intrauterine 0,05 ml of physiological solution on the 18th day of pregnancy. The antigen was split vaccine Vaxigrip 2009. Results and conclusion. In newborn animals, after intrauterine antigen action it was determined significantly increased content of PNA+ lymphocytes in the epithelium of gingival mucous, compared with control group, where PNA+ lymphocytes number gradually decreases. On the 11 th day of life, in animals of second group, quantity of intraepithelial PNA+ lymphocytes remains higher. On 45th day of postnatal formation its share does not significantly differ from similar indicators in all groups and decreases compared with neonatal period. Citation: Burega Yu. Distribution features of lymphocytes with peanut agglutinin positive receptors in gums epithelium of rats in norm and after intrauterine antigenic action. Morphologia. 2015;9(3:8-11.


    Marly Francisca Cândido


    Full Text Available

    Foram analisados 694 hemossoros de eqüinos, procedentes de sete rnunicípios goianos, encontrando-se uma positividade para aglutininas antileptóspira que variou de 8,8 a 25,0%, com o valor médio de 14,4%. Diferentes sorotipos foram pesquisados, pela técnica de GALTON et alii (1962, encontrando-se positividade para os sorotipos grippotyphosa, wolffi, ballum e australis. Os autores concluíram pela necessidade de prosseguimento de novas pesquisas, com a finalidade de averiguar-se a presença ou não de diferentes sorotipos observados em outros Estados e não encontrados na presente pesquisa, além da realização de estudo epidemiológico local e regional.

    694 equine serum blood tests were made from horses of 7 counties of the State of Goiás. Positive rates of Leptospira agglutinins varied from 8.8 to 25.0% with an average of 14.4%. Different serum types were studied by the technique developed by GALTON et alii (1962 finding positiveness for the following serum types: grippotyphosa, wolffi, ballum and australis.

  3. Enhanced Resistance of Snowdrop Lectin (Galanthus nivalis L. Agglutinin)-Expressing Maize to Asian Corn Borer (Ostrinia furnacalis Guenée)

    Zhao-Yu WANG; Xiao-Fen SUN; Fei WANG; Ke-Xuan TANG; Ju-Ren ZHANG


    In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-specific promoter were generated through the Agrobacterium tumefaciens-mediated also studied. Thirty-six independently derived plants were subjected to molecular analyses. The level of GNA expression at 0.13%-0.28% of total soluble protein was observed in different transgenic plants. The progeny of three GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to ACB. These plants synthesized GNA at levels above 0.24% total soluble protein and enhanced resistance to ACB was demonstrated by exposing the plants to insects under greenhouse conditions. Semi-artificial diet bioassays also showed the toxic effect of GNA on ACB. Field evaluation of the transgenic plants supported the results from the artificial trial. In the present study, we have obtained new insect-resistant maize material for further breeding work and have found that GNA-expressing plants not only gained significant resistance to homopterans, but also showed toxicity to ACB, which is a type of Lepidoptera.

  4. An Unusual Member of the Papain Superfamily: Mapping the Catalytic Cleft of the Marasmius oreades agglutinin (MOA) with a Caspase Inhibitor.

    Cordara, Gabriele; van Eerde, André; Grahn, Elin M; Winter, Harry C; Goldstein, Irwin J; Krengel, Ute


    Papain-like cysteine proteases (PLCPs) constitute the largest group of thiol-based protein degrading enzymes and are characterized by a highly conserved fold. They are found in bacteria, viruses, plants and animals and involved in a number of physiological and pathological processes, parasitic infections and host defense, making them interesting targets for drug design. The Marasmius oreades agglutinin (MOA) is a blood group B-specific fungal chimerolectin with calcium-dependent proteolytic activity. The proteolytic domain of MOA presents a unique structural arrangement, yet mimicking the main structural elements in known PLCPs. Here we present the X-ray crystal structure of MOA in complex with Z-VAD-fmk, an irreversible caspase inhibitor known to cross-react with PLCPs. The structural data allow modeling of the substrate binding geometry and mapping of the fundamental enzyme-substrate interactions. The new information consolidates MOA as a new, yet strongly atypical member of the papain superfamily. The reported complex is the first published structure of a PLCP in complex with the well characterized caspase inhibitor Z-VAD-fmk. PMID:26901797

  5. In vivo toxicity and immunogenicity of wheat germ agglutinin conjugated poly(ethylene glycol)-poly(lactic acid) nanoparticles for intranasal delivery to the brain

    Biodegradable polymer-based nanoparticles have been widely studied to deliver therapeutic agents to the brain after intranasal administration. However, knowledge as to the side effects of nanoparticle delivery system to the brain is limited. The aim of this study was to investigate the in vivo toxicity and immunogenicity of wheat germ agglutinin (WGA) conjugated poly(ethylene glycol)-poly(lactic acid) nanoparticles (WGA-NP) after intranasal instillation. Sprague-Dawley rats were intranasally given WGA-NP for 7 continuous days. Amino acid neurotransmitters, lactate dehydrogenase (LDH) activity, reduced glutathione (GSH), acetylcholine, acetylcholinesterase activity, tumor necrosis factor α (TNF-α) and interleukin-8 (IL-8) in rat olfactory bulb (OB) and brain were measured to estimate the in vivo toxicity of WGA-NP. Balb/C mice were intranasally immunized by WGA-NP and then WGA-specific antibodies in serum and nasal wash were detected by indirect ELISA. WGA-NP showed slight toxicity to brain tissue, as evidenced by increased glutamate level in rat brain and enhanced LDH activity in rat OB. No significant changes in acetylcholine level, acetylcholinesterase activity, GSH level, TNF-α level and IL-8 level were observed in rat OB and brain for the WGA-NP group. WGA-specific antibodies in mice serum and nasal wash were not increased after two intranasal immunizations of WGA-NP. These results demonstrate that WGA-NP is a safe carrier system for intranasal delivery of therapeutic agents to the brain.

  6. Topology of the polypeptide chain in the complex of agglutinin from castor bean seeds with β-D-galactose in the crystalline state

    The three-dimensional structure of the complex of agglutinin from Ricinus communis with β-D-galactose was established and refined at 2.5 A resolution by X-ray structure analysis. Biocrystals were obtained using dialysis through a semipermeable membrane. X-ray intensity data (Rmerge = 4.6%) were collected from one crystal at 100 K using synchrotron radiation at the DESY outstation [European Molecular Biology Laboratory (EMBL), Hamburg, Germany]. The initial phases were calculated by the molecular replacement method. The atoms of both protein and sugar molecules were localized. Unlike ricin, the ricinlike heterodimer RcA contains only one galactose-binding center in the region of the Asn46-Gly25-Trp37-Lys40 site in the first domain of the B subunit, whereas the second galactose-binding site of the B subunit is lost. One functionally important water molecule, which is bound to the residues Tyr123-Glu176-Arg179-Glu207, was revealed in the region of the active center in the A subunit

  7. Electrogenerated chemiluminescence biosensing for the detection of prostate PC-3 cancer cells incorporating antibody as capture probe and ruthenium complex-labelled wheat germ agglutinin as signal probe

    Yang, Haiying [Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi’an 710062 (China); Department of Chemistry, Yuncheng University, Yuncheng 044300 (China); Li, Zhejian; Shan, Meng; Li, Congcong; Qi, Honglan; Gao, Qiang [Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi’an 710062 (China); Wang, Jinyi [College of Science and College of Veterinary Medicine, Northwest A& F University, Yangling 712100 (China); Zhang, Chengxiao, E-mail: [Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi’an 710062 (China)


    Highlights: • A novel biosensor was developed for the detection of prostate cancer cells. • The selectivity of the biosensor was improved using antibody as capture probe. • The biosensor showed the low extremely detection limit of 2.6 × 10{sup 2} cells mL{sup −1}. • The ruthenium complex-labelled WGA can be transported in the cell vesicles. - Abstract: A highly selective and sensitive electrogenerated chemiluminescence (ECL) biosensor for the detection of prostate PC-3 cancer cells was designed using a prostate specific antibody as a capture probe and ruthenium complex-labelled wheat germ agglutinin as a signal probe. The ECL biosensor was fabricated by covalently immobilising the capture probe on a graphene oxide-coated glassy carbon electrode. Target PC-3 cells were selectively captured on the surface of the biosensor, and then, the signal probe was bound with the captured PC-3 cells to form a sandwich. In the presence of tripropylamine, the ECL intensity of the sandwich biosensor was logarithmically directly proportion to the concentration of PC-3 cells over a range from 7.0 × 10{sup 2} to 3.0 × 10{sup 4} cells mL{sup −1}, with a detection limit of 2.6 × 10{sup 2} cells mL{sup −1}. The ECL biosensor was also applied to detect prostate specific antigen with a detection limit of 0.1 ng mL{sup −1}. The high selectivity of the biosensor was demonstrated in comparison with that of a lectin-based biosensor. The strategy developed in this study may be a promising approach and could be extended to the design of ECL biosensors for highly sensitive and selective detection of other cancer-related cells or cancer biomarkers using different probes.

  8. Electrogenerated chemiluminescence biosensing for the detection of prostate PC-3 cancer cells incorporating antibody as capture probe and ruthenium complex-labelled wheat germ agglutinin as signal probe

    Highlights: • A novel biosensor was developed for the detection of prostate cancer cells. • The selectivity of the biosensor was improved using antibody as capture probe. • The biosensor showed the low extremely detection limit of 2.6 × 102 cells mL−1. • The ruthenium complex-labelled WGA can be transported in the cell vesicles. - Abstract: A highly selective and sensitive electrogenerated chemiluminescence (ECL) biosensor for the detection of prostate PC-3 cancer cells was designed using a prostate specific antibody as a capture probe and ruthenium complex-labelled wheat germ agglutinin as a signal probe. The ECL biosensor was fabricated by covalently immobilising the capture probe on a graphene oxide-coated glassy carbon electrode. Target PC-3 cells were selectively captured on the surface of the biosensor, and then, the signal probe was bound with the captured PC-3 cells to form a sandwich. In the presence of tripropylamine, the ECL intensity of the sandwich biosensor was logarithmically directly proportion to the concentration of PC-3 cells over a range from 7.0 × 102 to 3.0 × 104 cells mL−1, with a detection limit of 2.6 × 102 cells mL−1. The ECL biosensor was also applied to detect prostate specific antigen with a detection limit of 0.1 ng mL−1. The high selectivity of the biosensor was demonstrated in comparison with that of a lectin-based biosensor. The strategy developed in this study may be a promising approach and could be extended to the design of ECL biosensors for highly sensitive and selective detection of other cancer-related cells or cancer biomarkers using different probes

  9. Crystal Structure of the C-terminal Region of Streptococcus mutans Antigen I/II and Characterization of Salivary Agglutinin Adherence Domains

    Larson, Matthew R.; Rajashankar, Kanagalaghatta R.; Crowley, Paula J.; Kelly, Charles; Mitchell, Tim J.; Brady, L. Jeannine; Deivanayagam, Champion (King); (Cornell); (UAB); (Glasgow); (Florida)


    The Streptococcus mutans antigen I/II (AgI/II) is a cell surface-localized protein that adheres to salivary components and extracellular matrix molecules. Here we report the 2.5 {angstrom} resolution crystal structure of the complete C-terminal region of AgI/II. The C-terminal region is comprised of three major domains: C{sub 1}, C{sub 2}, and C{sub 3}. Each domain adopts a DE-variant IgG fold, with two {beta}-sheets whose A and F strands are linked through an intramolecular isopeptide bond. The adherence of the C-terminal AgI/II fragments to the putative tooth surface receptor salivary agglutinin (SAG), as monitored by surface plasmon resonance, indicated that the minimal region of binding was contained within the first and second DE-variant-IgG domains (C{sub 1} and C{sub 2}) of the C terminus. The minimal C-terminal region that could inhibit S. mutans adherence to SAG was also confirmed to be within the C{sub 1} and C{sub 2} domains. Competition experiments demonstrated that the C- and N-terminal regions of AgI/II adhere to distinct sites on SAG. A cleft formed at the intersection between these C{sub 1} and C{sub 2} domains bound glucose molecules from the cryo-protectant solution, revealing a putative binding site for its highly glycosylated receptor SAG. Finally, electron microscopy images confirmed the elongated structure of AgI/II and enabled building a composite tertiary model that encompasses its two distinct binding regions.

  10. Early Serologic Diagnosis of Mycoplasma pneumoniae Pneumonia: An Observational Study on Changes in Titers of Specific-IgM Antibodies and Cold Agglutinins.

    Lee, Sung-Churl; Youn, You-Sook; Rhim, Jung-Woo; Kang, Jin-Han; Lee, Kyung-Yil


    There have been some limitations on early diagnosis of Mycoplasma pneumoniae (MP) infection because of no immunoglobulin M (IgM) responses and variable detection rates of polymerase chain reaction in the early stage of the disease. We wanted to discuss regarding early diagnostic method using short-term paired titration of MP-specific IgM and cold agglutinins (CAs) in the early stage of MP pneumonia.The participants of this study were 418 children with MP pneumonia during 2 recent epidemics (2006-2007 and 2011), and they were diagnosed by an anti-MP IgM antibody test (Serodia Myco II) examined twice during hospitalization at presentation and around discharge (mean of 3.4 ± 1.3 days apart). CA titers were simultaneously examined twice during study period. Anti-MP IgM antibody titer ≥1:40 and CA titer ≥1:4 were considered positive, respectively. The relationships between 2 IgM antibodies in the early stage were evaluated.Regarding MP-specific antibody titers, 148 patients showed a seroconversion, 245 patients exhibited increased titers, and 25 patients had unchanged higher titers (≥1:640) during hospitalization. The median MP-specific antibody titers at each examination time were 1:80 and 1:640, respectively; those of CAs were 1:8 and 1:32, respectively. Illness duration prior to admission showed a trend of association with both titers, and patients with shorter illness duration had a higher rate of negative titers or lower titers at each examination time. CAs and MP-specific antibody titers were correlated in the total patients at presentation and at 2nd examination (P < 0.001, respectively), and the diagnostic corresponding rates of CAs to IgM antibody test were 81% to 96% in patient subgroups.Short-term paired MP specific-IgM determinations in the acute stage may be used as a definitive diagnostic method for MP pneumonia. Paired CA titers showed a correlation with MP-specific antibody titers, suggesting they can be used as an adjuvant diagnostic

  11. Expressions of Proliferating Cell Nuclear Antigen and Wheat Germ Agglutinin Receptor in Human Bladder Carcinoma%膀胱癌增殖细胞核抗原与麦胚凝集素受体的相关关系

    张士文; 葛根; 金伯涛


    [Purpose]To probe the relation of proliferating cell nuclear antigen (PCNA) and wheat germ agglutinin (WGA) receptors expressed in human bladder transitional cell carcinoma (TCC).[Methods]PCNA and WGA receptors were detected by immunohistochemical method (ABC method) in 63 specimens of TCC.[Results]We found that the distributions of PCNA and WGA receptors were increased with increase of histopathological grade in TCC (P<0.01).There was a higher expression in invasive tumors than that in superficial tumors (P<0.005),and there was a positive relation between PCNA and WGA receptors also.[Conclusion]It is shown that PCNA and WGA can be used as tumor markers for bladder cancer.%[目的 ]探讨增殖细胞核抗原 (proliferating cell nuclear antigen,PCNA)和麦胚凝集素 (wheat germ agglutinin,WGA)在膀胱移行细胞癌 (TCC)中表达的相关关系。 [方法 ]采用免疫组织化学 ABC法对 63例 TCC标本进行 PCNA和 WGA受体检测。 [结果 ]PCNA与 WGA的强阳性表达随着肿瘤的病理分级升高而增高;浸润性肿瘤中的 WGA受体的强阳性表达显著高于浅表性肿瘤 (P<0.05); PCNA与 WGA受体表达一致性良好,呈显著性相关 (P<0.005)。 [结论 ]我们认为 PCNA和 WGA受体均可作为 TCC的肿瘤标记物,证明了 TCC细胞的增殖活性增强将改变其细胞膜的抗原性。

  12. Expression of Vicia villosa agglutinin (VVA)-binding glycoprotein in primary breast cancer cells in relation to lymphatic metastasis: is atypical MUC1 bearing Tn antigen a receptor of VVA?

    Kawaguchi, Takanori; Takazawa, Hiroshi; Imai, Shunsuke; Morimoto, Junji; Watanabe, Takanori; Kanno, Masahiko; Igarashi, Seiji


    Aberrant carbohydrate expression frequently occurs in breast cancer and may endow cells with metastatic potential. Here we first studied the relationship between expression of Vicia villosa agglutinin (lectin) (VVA)-binding carbohydrates and aggressive breast cancer. We then investigated the molecular characteristics of these glycoproteins and compared them with those of glycoproteins recognized by the mouse anti-Tn monoclonal antibody (MAb) HB-Tn1. Histochemical studies of samples from 322 cases of invasive ductal carcinoma demonstrated that VVA-binding carbohydrate expression correlated with tumor stage, lymphatic invasion, and lymph node metastasis (p=0.0385, p=0.0019, and p=0.0430. respectively). Western blotting analysis of frozen materials from 39 cases, under denaturing and reducing conditions, revealed that the major cancer cell-specific VVA-binding proteins were molecules of about 30, 33, and >200 kDa. Cases expressing the approximately 33 kDa molecule had significant lymphatic invasion more frequently than did cases not expressing this molecule (p=0.0076). Binding of VVA to the approximately 30 and approximately 33 kDa molecules was completely lost by preincubation of VVA with 1 mM Tn antigen (N-acetylgalactosamine alpha1-O-serine). The VVA-binding molecules appeared to react with VU-3C6 anti-MUC1 MAb. Expression of HB-Tn1 in breast cancer cells showed significant correlation with expression of VVA-binding carbohydrate(s) (p<0.0001) but HB-Tn1 reactivity was not clearly related to breast cancer aggressiveness. Because anti-Tn MAbs bound to Tn antigen clusters, we concluded that atypical MUC1 bearing the noncluster form of Tn antigen is implicated in aggressive growth of primary breast cancer cells, particularly in lymphatic metastasis. PMID:16752227

  13. Aglutininas antileptospíricas em búfalos do Vale do Ribeira, Estado de São Paulo Anti-leptospire agglutinins in buffaloes from Vale do Ribeira, São Paulo State, Brazil

    Hélio Langoni


    Full Text Available Foram estudadas aglutininas antileptospíricas em 403 amostras de soro de búfalos, provenientes de sete Municípios do Vale do Ribeira, Estado de São Paulo, coletadas no período de janeiro de 1992 a junho de 1993. Utilizou-se o teste de soroaglutinação microscópica, considerando-se positivas as amostras cujo título fosse igual ou superior a 100. O maior título encontrado foi 1600 para o sorovar bratislava (1 amostra, seguido de 800 para wolffi (4 amostras. Do total, 152 (37,7% das amostras foram positivas, sendo que, dentre os sorovares testados, a prevalência em ordem decrescente foi: wolffi (68, 44,8%, icterohaemorrhagiae (51, 33,6%, hardjo (51, 33,6%, castellonis (25, 16,5%, djasiman (12, 7,9%, grippotyphosa (10, 6,6%, pomona (8, 5,2%, bratislava (6, 4,0%, copenhageni (5, 3,3% e tarassovi (4, 2,7%.A total of 403 buffaloes serum samples from seven counties of Vale do Ribeira, São Paulo State, Brazil, obtained between January 1992 and June 1993, were studied to determine the prevalence of anti-leptospire agglutinins using the microscopic serum agglutination test. A titre of 100 and above was considered positive. The highest titre found was 1600 to the sorovar bratislava (one sample, followed by 800 to wolffi (4 samples. 152 (37.7% samples from the total were positive to the serovars tested, and their prevalence, in decreasing order, was: wolffi (68, 44.8%, icterohaemorrhagiae (51, 33.6%, hardjo (51, 33.6%, castellonis (25, 16.5%, djasiman (12, 7.9%, grippotyphosa (10, 6.6%, pomona (8, 5.2%, bratislava (6, 4.0%, copenhageni (5, 3.3% and tarassovi (4, 2.7%.

  14. Cold agglutinin disease (CADwith autoimmune haemolytic anaemia: a case report of a coronary artery disease patient Doença por aglutininas a frio (DAC com anemia hemolítica auto-imune: relato de caso de um coronariopata

    Leandro A. Barbosa


    Full Text Available Cold agglutinin disease (CAD with autoimmune haemolytic anemia is characterized by the production of harmful cold autoantibodies associated with increased red cell destruction during exposure to cold. The treatment of CAD is very difficult and a great effort is required to obtain therapeutic success. Cyclophosphamide is a potent immunosuppressive agent which is widely used in all bone marrow transplantation conditioning regimens for patients with acquired severe aplastic anemia. In this report, we describe the case of a coronary artery disease patient with severe CAD, but without lymphoproliferative disease, in which general measures and immunosuppressive therapies were adopted, there by avoiding blood transfusions.A doença por aglutininas a frio (CAD cursando com anemia hemolítica auto-imune (AHAI é decorrente da produção de autoanticorpos que reagem muito bem a baixas temperaturas, dirigidos contra hemácias autólogas. A habilidade desses anticorpos em destruir as hemácias encontra-se diretamente relacionada à sua capacidade em fixar complemento durante a exposição do paciente a baixas temperaturas. A AHAI por anticorpos frios pode ser idiopática - ausência de doença de base - ou secundária, geralmente associada a desordens linfoproliferativas de células B ou determinados processos infecciosos. A hemólise é intravascular, através de aglutininas da classe IgM, com teste direto da antiglobulina humana positivo para complemento. O tratamento da CAD é difícil, exigindo um esforço contínuo, necessário para se obter sucesso terapêutico. A ciclofosfamida é um agente imunossupressor potente, amplamente utilizado em transplantes de medula óssea, particularmente nos portadores de anemia aplástica. Descrevemos o caso de um coronariopata portador de CAD severa, cuja exploração diagnóstica excluiu doença linfoproliferativa. Adotamos medidas gerais de suporte e terapia imunossupressora, coibindo o uso de hemotransfusões.

  15. Determination of trace alkaline phosphatase by affinity adsorption solid substrate room temperature phosphorimetry based on wheat germ agglutinin labeled with 8-quinolineboronic acid phosphorescent molecular switch and prediction of diseases

    Liu, Jia-Ming; Gao, Hui; Li, Fei-Ming; Shi, Xiu-Mei; Lin, Chang-Qing; Lin, Li-Ping; Wang, Xin-Xing; Li, Zhi-Ming


    The 8-quinolineboronic acid phosphorescent molecular switch (abbreviated as PMS-8-QBA. Thereinto, 8-QBA is 8-quinolineboronic acid, and PMS is phosphorescent molecular switch) was found for the first time. PMS-8-QBA, which was in the "off" state, could only emit weak room temperature phosphorescence (RTP) on the acetyl cellulose membrane (ACM). However, PMS-8-QBA turned "on" automatically for its changed structure, causing that the RTP of 8-QBA in the system increased, after PMS-8-QBA-WGA (WGA is wheat germ agglutinin) was formed by reaction between -OH of PMS-8-QBA and -COOH of WGA. More interesting is that the -NH 2 of PMS-8-QBA-WGA could react with the -COOH of alkaline phosphatase (AP) to form the affinity adsorption (AA) product WGA-AP-WGA-8-QBA-PMS (containing -NH-CO- bond), which caused RTP of the system to greatly increase. Thus, affinity adsorption solid substrate room temperature phosphorimetry using PMS-8-QBA as labelling reagent (PMS-8-QBA-AA-SSRTP) for the determination of trace AP was established. The method had many advantages, such as high sensitivity (the detection limit (LD) was 2.5 zg spot -1. For sample volume of 0.40 μl spot -1, corresponding concentration was 6.2 × 10 -18 g ml -1), good selectivity (the allowed concentration of coexisting material was higher, when the relative error was ±5%), high accuracy (applied to detection of AP content in serum samples, the result was coincided with those obtained by enzyme-linked immunoassay), which was suitable for the detection of trace AP content in serum samples and the forecast of human diseases. Meanwhile, the mechanism of PMS-8-QBA-AASSRTP was discussed. The new field of analytical application and clinic diagnosis technique of molecule switch are exploited, based on the phosphorescence characteristic of PMS-8-QBA, the AA reaction between WGA and AP, as well as the relation between AP content and human diseases. The research results promote the development and interpenetrate among molecule


    Vânia Lúcia de Assis Santana


    Full Text Available Objetivou-se investigar a freqüência de aglutininas anti-Brucella abortus em caprinos e ovinos do Sertão do Estado de Pernambuco, Brasil. Foram processadas 700 amostras de soros sangüíneos, das quais 340 eram da espécie caprina (115 machos e 225 fêmeas e 360 (136 machos e 224 fêmeas ovina. Empregou-se a técnica do antígeno acidificado tamponado (AAT corado com rosa bengala (RB. Das 340 amostras de caprinos avaliadas, duas (0,6% foram reagentes ao AAT. Não se observaram associações significativas para as variáveis faixa etária (p= 0,430, raça (p= 0,936 e sexo (p= 0,562. Das 360 amostras de ovinos, nove (2,5% foram reagentes. Também não houve associação significativa entre as variáveis analisadas e a soropositividade para brucelose: faixa etária (p= 0,522; raça (p= 0,576 e sexo (p= 0,461. Verificou-se associação significativa (p= 0,042 entre as espécies estudadas e soropositividade para brucelose nos animais investigados. A soropositividade para Brucella abortus em caprinos e ovinos foi descrita pela primeira vez no Sertão de Pernambuco, fato que pode dificultar o sucesso do Programa Nacional de Controle e Erradicação da Brucelose, tendo em vista que nessa região é comum a criação consorciada de pequenos ruminantes com bovinos, além de representar riscos à Saúde Pública.

    PALAVRAS-CHAVES: Brucelose, ovinos, caprinos, pequenos ruminantes, sorodiagnóstico. The objective was to investigate the frequency of anti-Brucella abortus agglutinins in goats and sheep of the backlands of the State of Pernambuco, Brazil. 700 samples of sanguine serums were processed, of which 340 were of the goat (115 males and 225 females and 360 (136 males and 224 females sheep. The technique of the Tamponed Acidified Antigen (AAT dyed with Bengalese Rose (BR was used. Of the 340 samples of goat evaluated two (0.6% were reactive to AAT. Significant associations were not observed for the variable age group (p = 0.430; race (p = 0

  17. Diversidade antigênica entre amostras de Arcobacter spp isoladas de suínos no Rio Grande do Sul e presença de anticorpos aglutinantes em amostras de soro de porcas com problemas reprodutivos Antigenic diversity among strains of Arcobacter spp isolated from pigs in Rio Grande do Sul, Brazil and presence of agglutinin titers in serum samples of sows with reproductive problems

    Sérgio José de Oliveira


    there was presence of agglutinins in serum samples.

  18. Overexpression of glycosylated proteins in cervical cancer recognized by the Machaerocereus eruca agglutinin Overexpression of glycosylated proteins in cervical cancer recognized by the Machaerocereus eruca agglutinin

    Carlos Solórzano


    Full Text Available In cervical cancer, glycosylation has been suggested as being involved in both its carcinogenesis and
    invasive capacity. In this work, we analyzed mucin type O-glycosylation in biopsies of invasive cervical cancer in
    FIGO stage II B through histochemistry using lectins specific for O-glycosidically linked glycans. Our results
    reveal that the lectin Machaerocereus eruca (MeA, specific for Gal in a Fuca1,2 (GalNAca1,3 Galb1,4 showed
    increased recognition of tumoral cells and tumoral stroma tissue compared to other lectins with similar specificity;
    healthy cervical tissue was negative for MeA. Trypsin treatment of recognized tissues abolished MeA’s recognition;
    moreover, interaction of MeA was inhibited with oligosaccharides from mucin. As demonstrated by
    Western blot of 2-D electrophoresis, MeA recognized ten glycoproteins in the range from 122 to 42 kDa in
    cervical cancer lysates. The LC-ESI-MS/MS analysis of the MeAs’ recognized peptides revealed that the latter
    matched mainly with the amino acid sequences of lamin A/C, vimentin, elongation factor 2, keratin 1, and beta
    actin. Our results suggest that MeA recognizes a complex of over-expressed O-glycosidically-linked proteins that
    play a relevant role in cervical cancer’s invasive capacity. O-glycosylation participates in the disassembly of intercellular
    junctions favoring cancer progression.In cervical cancer, glycosylation has been suggested as being involved in both its carcinogenesis and
    invasive capacity. In this work, we analyzed mucin type O-glycosylation in biopsies of invasive cervical cancer in
    FIGO stage II B through histochemistry using lectins specific for O-glycosidically linked glycans. Our results
    reveal that the lectin Machaerocereus eruca (MeA, specific for Gal in a Fuca1,2 (GalNAca1,3 Galb1,4 showed
    increased recognition of tumoral cells and tumoral stroma tissue compared to other lectins with similar specificity;
    healthy cervical tissue was negative for MeA. Trypsin treatment of recognized tissues abolished MeA’s recognition;
    moreover, interaction of MeA was inhibited with oligosaccharides from mucin. As demonstrated by
    Western blot of 2-D electrophoresis, MeA recognized ten glycoproteins in the range from 122 to 42 kDa in
    cervical cancer lysates. The LC-ESI-MS/MS analysis of the MeAs’ recognized peptides revealed that the latter
    matched mainly with the amino acid sequences of lamin A/C, vimentin, elongation factor 2, keratin 1, and beta
    actin. Our results suggest that MeA recognizes a complex of over-expressed O-glycosidically-linked proteins that
    play a relevant role in cervical cancer’s invasive capacity. O-glycosylation participates in the disassembly of intercellular
    junctions favoring cancer progression.

  19. Neuroglial response to neuron injury. A study using intraneural injection of ricinus communis agglutinin-60.

    Ling, E A; Wen, C Y; J.Y. Shieh; Yick, T Y; Leong, S K


    The present study has shown the selective destruction of large ventral horn neurons in the lumbosacral cord segments following a single injection of RCA-60 into the sciatic nerve. The neurons appeared to undergo structural alteration beginning 3 days after the RCA application. In the postoperative period extending from 1 to 60 days, degeneration of neurons was progressive and irreversible and this elicited a rapid increase in the number of microglial cells. They were most numerous in the 7 da...

  20. Neuroglial response to neuron injury. A study using intraneural injection of ricinus communis agglutinin-60.

    Ling, E A; Wen, C Y; Shieh, J Y; Yick, T Y; Leong, S K


    The present study has shown the selective destruction of large ventral horn neurons in the lumbosacral cord segments following a single injection of RCA-60 into the sciatic nerve. The neurons appeared to undergo structural alteration beginning 3 days after the RCA application. In the postoperative period extending from 1 to 60 days, degeneration of neurons was progressive and irreversible and this elicited a rapid increase in the number of microglial cells. They were most numerous in the 7 days postoperative animals. The massive microglial cells penetrated the neuropil and appeared to strip off the axon terminals from the postsynaptic somata. Occasional axon terminals were phagocytosed by microglia. The numerous microglial cells often formed a multilayered 'barrier' encircling the somata of the RCA-poisoned neurons which eventually became totally disorganised. It is postulated that in the course of neuronal degeneration induced by RCA, microglial cells serve to prevent the leakage or diffusion of the toxic lectin from the neuronal somata into the neighbouring neuropil. They also function as scavenger cells in the removal of degenerating myelinated axons in the longer surviving rats. Oligodendrocytes do not appear to react actively to the degeneration process. However, astrocytes showed a significant increase in the 7 and 15 day postoperative rats and this coincided with the presence of mitotic astrocytes in the same period. PMID:2606792

  1. Ulex europaeus agglutinin II (UEA-II) is a novel, potent inhibitor of complement activation

    Lekowski, Robert; Collard, Charles D.; Reenstra, Wende R.; Stahl, Gregory L.


    Complement is an important mediator of vascular injury following oxidative stress. We recently demonstrated that complement activation following endothelial oxidative stress is mediated by mannose-binding lectin (MBL) and activation of the lectin complement pathway. Here, we investigated whether nine plant lectins which have a binding profile similar to that of MBL competitively inhibit MBL deposition and subsequent complement activation following human umbilical vein endothelial cell (HUVEC)...

  2. Anti-leptospiral agglutinins in marmosets (Saguinus oedipus and Saguinus leucopus from illegal trade

    Viviana Gonzalez-Astudillo


    Full Text Available Objective. Determine the infection status with pathogenic Leptospira of one Saguinus oedipus and nine Saguinus leucopus at the Cali Zoo that had been confiscated in Colombia from illegal trade. Materials and methods. A full physical examination, blood work, urinalysis were conducted in all individuals during the reception health check-up, in addition to running the microagglutination test with a pool of 19 serovars, with a starting dilution of 1:50. Results. A high positive titer (≥1:3200 to Leptospira alexanderi serovar manhao in an asymptomatic S. oedipus was detected. All S. leucopus tested negative or less than 1:50. Conclusions. Captive locations have been documented to artificially enhance opportunities to come into contact with contaminated bodily fluids from peridomestic rodents. However, infectious diseases acquired during the illegal transport of wildlife to major metropolitan centers are rarely considered a wildlife conservation or public health threat. Infection with zoonotic pathogens should also be considered an additional threat to endangered wild primates involved in illegal trade, which could hamper reintroduction efforts or other population management procedures for primate species with restricted and fragmented distributions.

  3. Dynamics of Agglutinin-Like Sequence (ALS) Protein Localization on the Surface of Candida Albicans

    Coleman, David Andrew


    The ALS gene family encodes large cell-surface glycoproteins associated with "C. albicans" pathogenesis. Als proteins are thought to act as adhesin molecules binding to host tissues. Wide variation in expression levels among the ALS genes exists and is related to cell morphology and environmental conditions. "ALS1," "ALS3," and "ALS4" are three of…

  4. Leptospiral agglutinins in captive and free ranging non-human primates in Sarawak, Malaysia

    S. Thayaparan


    Full Text Available Aim: The proposed study was carried out to determine the extent of exposure to leptospirosis in non-human primates. Materials and Methods: Trapping of non-human primates was carried out opportunistically around the Bako National Park and the Matang Wildlife Center in the vicinity of human settlements and tourism areas of Sarawak. Blood samples were obtained from the saphenous vein to determine the presence of antibodies by the Microscopic Agglutination Test (MAT to 17 serovars of Leptospira commonly found in Malaysia. Results: This study reports the screening of twelve primates (eight captive and four free ranging for leptospirosis. Eight of the 12 monkeys (66.6%; 95% CI 34.9-90.1 reacted against one or two serovars of Leptospira (Lai and Leptospira Lepto175. The serovar Lai is considered pathogenic for different mammals, including humans. Leptospira Lepto 175 has been identified as an intermediate strain and further studies are being undertaken on this serovar. Conclusion: These results are important as primates may act as reservoirs of Leptospira spp. for humans, which may potentially affect tourism (economic loss, conservation efforts and public health.

  5. Postmitotic basal cells in squamous cell epithelia are identified with Dolichos biflorus agglutinin - functional consequences

    Hrdličková-Celá, E.; Plzák, J.; Holíková, Z.; Dvořánková, B.; Smetana, Karel


    Roč. 109, č. 10 (2001), s. 714-720. ISSN 0903-4641 R&D Projects: GA AV ČR KSK4055109 Institutional research plan: CEZ:AV0Z4050913 Keywords : squamous cell epithelia * carcinoma * lectin Subject RIV: CE - Biochemistry Impact factor: 1.924, year: 2001

  6. Wisteria Floribunda Agglutinin-Labeled Perineuronal Nets in the Mouse Inferior Colliculus, Thalamic Reticular Nucleus and Auditory Cortex

    Sarah M. Fader; Kazuo Imaizumi; Yuchio Yanagawa; Lee, Charles C.


    Perineuronal nets (PNNs) are specialized extracellular matrix molecules that are associated with the closing of the critical period, among other functions. In the adult brain, PNNs surround specific types of neurons, however the expression of PNNs in the auditory system of the mouse, particularly at the level of the midbrain and forebrain, has not been fully described. In addition, the association of PNNs with excitatory and inhibitory cell types in these structures remains unknown. Therefore...

  7. Transsynaptic transport of wheat germ agglutinin expressed in a subset of type II taste cells of transgenic mice

    Mosinger Bedrich


    Full Text Available Abstract Background Anatomical tracing of neural circuits originating from specific subsets of taste receptor cells may shed light on interactions between taste cells within the taste bud and taste cell-to nerve interactions. It is unclear for example, if activation of type II cells leads to direct activation of the gustatory nerves, or whether the information is relayed through type III cells. To determine how WGA produced in T1r3-expressing taste cells is transported into gustatory neurons, transgenic mice expressing WGA-IRES-GFP driven by the T1r3 promoter were generated. Results Immunohistochemistry showed co-expression of WGA, GFP and endogenous T1r3 in the taste bud cells of transgenic mice: the only taste cells immunoreactive for WGA were the T1r3-expressing cells. The WGA antibody also stained intragemmal nerves. WGA, but not GFP immunoreactivity was found in the geniculate and petrosal ganglia of transgenic mice, indicating that WGA was transported across synapses. WGA immunoreactivity was also found in the trigeminal ganglion, suggesting that T1r3-expressing cells make synapses with trigeminal neurons. In the medulla, WGA was detected in the nucleus of the solitary tract but also in the nucleus ambiguus, the vestibular nucleus, the trigeminal nucleus and in the gigantocellular reticular nucleus. WGA was not detected in the parabrachial nucleus, or the gustatory cortex. Conclusion These results show the usefulness of genetically encoded WGA as a tracer for the first and second order neurons that innervate a subset of taste cells, but not for higher order neurons, and demonstrate that the main route of output from type II taste cells is the gustatory neuron, not the type III cells.

  8. Immune haemolytic anaemia associated with ampicillin dependent warm antibodies and high titre cold agglutinins in a patient with Mycoplasma pneumonia

    Mickley, H; Sørensen, P G


    A case of severe immune haemolytic anaemia in a 54-year-old man suffering from Mycoplasma pneumonia is presented. A strongly positive direct Coombs test with erythrocyte bound IgG, C3d and C4 was demonstrated during the haemolytic process. Further, serologic investigations revealed ampicillin-dep...

  9. Endogenous lectins from cultured soybean cells: isolation of a protein immunologically cross-reactive with seed soybean agglutinin and analysis of its role in binding of Rhizobium japonicum


    Incubation of Rhizobium japonicum with the cultured soybean cell line SB-1, originally derived from the roots of Glycine max, resulted in specific adhesion of the bacteria to the plant cells. This binding interaction appears to be mediated via carbohydrate recognition, since galactose can inhibit the heterotypic adhesion but glucose cannot. Affinity chromatography, on a Sepharose column derivatized with N- caproyl-galactosamine, of the supernatant fraction of a SB-1 cell suspension after enzy...

  10. Ultrastructural changes of the nodose ganglion cells following an intraneural injection of Ricinus communis agglutinin-60 into the vagus nerve in hamsters.

    Ling, E A; Wen, C Y; Shieh, J Y; Yick, T Y; Wong, W C


    Virtually all the ganglion cells in the nodose ganglion in hamsters underwent rapid degeneration following an intraneural injection of RCA-60 into the vagus nerve in the cervical region. The earliest signs of neuronal degeneration were evident in animals which survived 5 days after the ricin application. A remarkable feature was the appearance of a variable number of granular dense bodies measuring 1-4 microns in diameter in the cytoplasm. They were composed of closely stacked cisternae which...

  11. Luffa acutangula agglutinin: Primary structure determination and identification of a tryptophan residue involved in its carbohydrate-binding activity using mass spectrometry.

    Kumar, Gnanesh; Mishra, Padmanabh; Anantharam, Vellareddy; Surolia, Avadhesha


    A lectin from phloem exudates of Luffa acutangula (ridge gourd) was purified on chitin affinity chromatography and characterized for its amino acid sequence and to study the role of tryptophan in its activity. The purified lectin was subjected to various proteolytic digestions, and the resulting peptides were analyzed by liquid chromatography coupled electrospray ionization ion trap mass spectrometer. The peptide precursor ions were fragmented by collision-induced dissociation or electron transfer dissociation experiments, and a manual interpretation of MS/MS was performed to deduce amino acid sequence. This gave rise to almost complete sequence coverage of the lectin which showed high-sequence similarity with deduced sequences of phloem lectins present in the database. Chemical modification of lysine, tyrosine, histidine, arginine, aspartic acid, and glutamic acid residues did not inhibit the hemagglutinating activity. However, the modification of tryptophan residues using N-bromosuccinimide showed the loss of hemagglutinating activity. Additionally, the mapping of tryptophan residues was performed to determine the extent and number of residues modified, which revealed that six residues per molecule were oxidized suggesting their accessibility. The retention of the lectin activity was seen when the modifications were performed in the presence of chitooligosaccharides due to protection of a tryptophan residue (W102) in the protein. These studies taken together have led to the identification of a particular tryptophan residue (W102) in the activity of the lectin. PMID:26597132

  12. Cell surface display of highly pathogenic avian influenza hemagglutinin on the surface of Pichia pastoris cells using alpha-agglutinin for production of oral vaccines

    Yeast are an ideal organism to express viral antigens because yeast glycosylate proteins are more similar to mammals than bacteria, and expression of proteins in yeast is relatively fast and inexpensive. In addition to the convenience of production, for purposes of vaccination, yeast have been show...

  13. Anti-leptospirose agglutinins in equine sera, from São Paulo, Goias, and Mato Grosso do Sul, Brazil, 1996-2001

    H. Langoni


    Full Text Available Equine leptospirosis can present a non-symptomatic form, an acute clinical form, or even develop chronically, causing reproductive alterations, such as abortion and recurrent uveitis. Since the prevalence of leptospirosis in several countries and regions is widely reported, the objective of this study was to verify the prevailing equine leptospirosis in different regions of Brazil. Sera from 1402 blood samples from horses of different age, sex, breed, and purpose were examined. These samples came from southeastern and central west states of Brazil. The method utilized was the Microscopic Agglutination Test (MAT, with 12 different Leptospira serovars. From the sera tested, 754 (54% were positive for one (385 or more (372 serovars. These results were higher when compared to national and international levels. The most commonly found serovars were icterohaemorrhagiae (37.01%, suggesting exposure to rodents, castellonis (16.97%, and djasiman (15.19%. There were significant differences of reagents between sexes, and a tendency toward higher positivity with age. Distribution of sera-reagents related to aptitude showed a markedly higher value for work animals than for sporting ones. Higher rates were found for animals with undefined breed. There were no significant differences related to regional origin. As an indication of multiple exposure, significant associations were observed between the following serovars: castellonis and djasiman; castellonis and grippotyphosa; castellonis and copenhageni; castellonis and icterohaemorrhagiae; castellonis and pomona; canicola and pomona; canicola and djasiman; djasiman and copenhageni; icterohaemorrhagiae and djasiman; icterohaemorrhagiae and pyrogenes; copenhageni and pomona. These results showed the necessity of further studies on the epidemiology of this disease in equines and its relationship to human illness.

  14. Main: 2CWG [RPSD[Archive

    Full Text Available 2CWG 小麦 Bread Wheat ... Triticum aestivum Agglutinin Isolectin 1 Precursor Triticum Aestivum Wheat ... efinement And Structure Analysis Of The Complex Of Wheat ... Germ Agglutinin With A Bivalent Sialoglycopeptide ... GGRVCTNNYCCSKWGSCGIGPGYCGAGCQSGGCDGVFAEAITANSTLLQE wheat _2CWG.jpg ...

  15. How Is Waldenstrom Macroglobulinemia Diagnosed?

    ... in cool temperatures and can block blood vessels). Cold agglutinins Cold agglutinins are antibodies that attack and ... attach to cells only if they contain specific molecules. These antibodies cause color changes, which can be ...

  16. Salivary agglutinin and lung scavenger receptor cysteine-rich glycoprotein 340 have broad anti-influenza activities and interactions with surfactant protein D that vary according to donor source and sialylation

    Hartshorn, Kevan L.; Ligtenberg, Antoon; White, Mitchell R.; van Eijk, Martin; Hartshorn, Max; Pemberton, Lily; Holmskov, Uffe; Crouch, Erika


    salivary gp-340 are identical in protein sequence, salivary gp-340 from one donor had significantly greater antiviral activity against avian-like IAV strains which preferentially bind sialic acids in alpha(2,3) linkage. A greater density of alpha(2,3)-linked sialic acids was present on the salivary gp-340...... from this donor as compared with salivary gp-340 from another donor or several preparations of lung gp-340. Hence, the specificity of sialic acid linkages on gp-340 is an important determinant of anti-IAV activity. Gp-340 binds to SP-D (surfactant protein D), and we previously showed that lung gp-340...

  17. Pesquisa de aglutininas anti Brucella canis em soros humanos na cidade de São Paulo, Brasil Research on agglutinins for Brucella canis in human sera in the city of S. Paulo, Brazil

    Maria Helena Matiko Akao Larsson


    Full Text Available De 330 soros humanos examinados pela prova de soroaglutinação lenta em tubos, 4(1,21% apresentaram aglutininas anti Brucella canis em diluição 1:100 (1 reagente com título 100, 2 reagentes com título 200 e 1 reagente com título 400.Of the 330 human sera tested by tube agglutination test, 4 (1.21% were positive for Brucella canis antibodies with tilers 1:100 or higher (1 reagent with titer of 1:100, 2 reagents with titer of 1:200, and 1 reagent with tiler of 1:400.

  18. 小扁豆凝集素结合型甲胎异质体在肝癌诊断中的意义%Significance of Lens culinaris agglutinin-reactive alpha-fetoprotein in the diagnosis of hepatocellular carcinoma

    卓传尚; 柳丽娟; 吴秋芳


    目的 探讨小扁豆凝集素结合型甲胎蛋白异质体(AFP-L3)在良恶性肝病鉴别诊断的临床价值.方法 应用装有耦联了小扁豆凝集素(LCA)的微量离心柱分离185例肝病患者的AFP-L3,用时间分辨荧光免疫检测血清AFP和AFP-L3含量,计算AFP-L3%.结果 肝细胞癌患者的AFP-L3%明显高于其他肝病患者(χ2=29.329,P<0.001);AFP、AFP-L3%检测肝细胞癌在ROC曲线下的面积AUC分别为0.407和0.841;以AFP-L3%≥12.6%作为诊断标准,AFP-L3诊断肝细胞癌敏感性和特异性分别为83.3%和86.3%.结论 AFP-L3对肝细胞癌诊断准确度明显高于AFP,微量离心柱法检测AFP-L3在良恶性肝脏病变鉴别诊断中具有重要临床价值.

  19. 阴道或直肠注入白芸豆凝集素的长期毒性研究%Study on long-term toxicity test of Yunnan white kidney bean agglutinin in mice

    赵琎; 朱轩轩; 李阔; 杨明洁; 张田; 王敏康


    目的 观察长期阴道或直肠注入白芸豆植物凝集素( WKBA)对小鼠的影响,为人用安全剂量提供参考.方法 将昆明种小鼠分成白芸豆凝集素高、中、低剂量组、壬苯醇醚(乐乐迷,Nonoxynol -9)组、辅料组、生理盐水组,每组雌雄各15只.分别采用阴道/肛门给药.1周给药5次.3个月后进行检查.结果 连续3个月阴道或直肠注入白芸豆凝集素( ≤4 mg/d),小鼠的日常生活没有异常,对小鼠的尿液没有影响,而小鼠的血液学和血液生化指标没有变化规律.结论 连续3个月阴道或直肠注入白芸豆凝集素(≤4 mg/d),小鼠的日常生活没有异常,对小鼠的尿液指标和脏器系数没有影响,而对小鼠血液的生化指标影响不明确.

  20. Correlation between carbohydrate structures on the envelope glycoprotein gp120 of HIV-1 and HIV-2 and syncytium inhibition with lectins

    Hansen, J E; Nielsen, C M; Nielsen, C;


    The binding of 13 different lectins to gp120 partially purified from two HIV-1 isolates and one HIV-2 isolate was studied by in situ staining on electrophoretically separated and electroblotted HIV antigens. The lectins concanavalin A, wheat germ agglutinin, Lens culinaris agglutinin, Vicia faba...... agglutinin, Pisum sativum agglutinin and phytohaem(erythro)agglutinin bound to gp120 of all three isolates. The carbohydrate of gp120 recognized by lectins was thus arranged in at least four types of glycans: a high mannose type glycan, a bisected hybrid or complex type glycan, a biantennary fucosylated...

  1. Glycoproteins from the cuticle of the Atlantic shore crab Carcinus maenas: I. Electrophoresis and Western-blot analysis by use of lectins

    Compère, P.; Jaspar-Versali, M.-F.; Goffinet, G.


    The protein and glycoprotein content of four different neutral or acidic solvent extracts (0.5 M KCl, 10% EDTA, 0.1 N HCI, or 2% acetic acid) from the mineralized exoskeleton of a decapod crustacean, the Atlantic shore crab Carcinus maenas, were characterized by quantitative analysis of proteins, SDS-PAGE analysis, and probing with lectins on blots. The lectins used were Conconavalin A, Jacalin, soybean agglutinin, Maackia amurensis agglutinin II, and Sambucus nigra agglutinin. The results sh...

  2. NCBI nr-aa BLAST: CBRC-MLUC-01-0257 [SEVENS

    Full Text Available Complexes pdb|2CWG|A Chain A, Crystallographic Refinement And Structure Analysis ...inement And Structure Analysis Of The Complex Of Wheat Germ Agglutinin With A Bival...Of The Complex Of Wheat Germ Agglutinin With A Bivalent Sialoglycopeptide From Glycophorin A pdb|2CWG|B Chain B, Crystallographic Ref

  3. NCBI nr-aa BLAST: CBRC-MLUC-01-0257 [SEVENS

    Full Text Available CBRC-MLUC-01-0257 sp|P10969|AGI3_WHEAT ... RecName: Full=Agglutinin isolectin 3; AltName: Full=WGA3; ... Flags: Precursor gb|AAA34257.1| wheat ... germ agglutinin [Triticum turgidum subsp. durum] p ...

  4. NCBI nr-aa BLAST: CBRC-MLUC-01-0257 [SEVENS

    Full Text Available CBRC-MLUC-01-0257 pdb|2UWZ|A Chain A, Crystal Structure Of Wheat ... Germ Agglutinin Isolectin 3 In ... te Ligand pdb|2UWZ|B Chain B, Crystal Structure Of Wheat ... Germ Agglutinin Isolectin 3 In Complex With A Synt ...

  5. Main: 1WGC [RPSD[Archive

    Full Text Available 1WGC 小麦 Bread Wheat ... Triticum aestivum Agglutinin Isolectin 1 Precursor Triticum Aestivum Wheat ... Analysis Of Two Refined N-Acetylneuraminyllactose-*Wheat ... Germ Agglutinin Isolectin Complexes J.Mol.Biol. V. ... GGRVCTNNYCCSKWGSCGIGPGYCGAGCQSGGCDGVFAEAITANSTLLQE wheat _1WGC.jpg ...

  6. Main: 2WGC [RPSD[Archive

    Full Text Available 2WGC 小麦 Bread Wheat ... Triticum aestivum Agglutinin Isolectin 2 Precursor Triticum Aestivum Wheat ... Analysis Of Two Refined N-Acetylneuraminyllactose-*Wheat ... Germ Agglutinin Isolectin Complexes J.Mol.Biol. V. ... GGRVCTNNYCCSKWGSCGIGPGYCGAGCQSGGCDAVFAGAITANSTLLAE wheat _2WGC.jpg ...

  7. Lectin staining shows no evidence of involvement of glycocalyx/mucous layer carbohydrate structures in development of celiac disease

    Toft-Hansen, Henrik; Nielsen, Christian; Biagini, Matteo;


    CD and T1D were examined before and after remission following a gluten-free diet. We performed lectin histochemistry using peanut agglutinin (PNA) and Ulex europaeus agglutinin (UEA) staining for Gal-β(1,3)-GalNAc and Fucα1-2Gal-R, respectively, of the glycocalyx/mucous layer. The staining was scored...

  8. Histological and lectin histochemical studies on the olfactory and respiratory mucosae of the sheep.

    Ibrahim, Dalia; Nakamuta, Nobuaki; Taniguchi, Kazumi; Yamamoto, Yoshio; Taniguchi, Kazuyuki


    The olfactory and respiratory mucosae of the Corriedale sheep were examined using lectin histochemistry in order to clarify the histochemical and glycohistochemical differences between these two tissues. The olfactory epithelium was stained with 13 lectins out of 21 lectins examined, while the respiratory epithelium was positive to 16 lectins. The free border of both of the olfactory and respiratory epithelia was stained with 12 lectins: Wheat germ agglutinin (WGA), succinylated-wheat germ agglutinin (s-WGA), Lycopersicon esculentum lectin (LEL), Solanum tuberosum lectin (STL), Datura stramonium lectin (DSL), Soybean agglutinin (SBA), Bandeiraea simplicifolia lectin-I (BSL-I), Ricinus communis agglutinin-I (RCA-120), Erythrina cristagalli lectin (ECL), Concanavalin A (Con A), Phaseolus vulgaris agglutinin-E (PHA-E) and Phaseolus vulgaris agglutinin-L (PHA-L). The associated glands of the olfactory mucosa, Bowman's glands, were stained with 13 lectins. While both the goblet cells and mucous nasal glands were stained with 8 lectins; five of them (WGA, s-WGA, STL, Vicia villosa agglutinin (VVA) and ECL) were mutually positive among the Bowman's glands, mucous nasal glands and the goblet cells. These findings indicate that the glycohistochemical characteristics of the free borders of both olfactory and respiratory epithelia are similar to each other, suggesting that secretions from the Bowman's glands and those of the goblet cells and mucous nasal glands are partially exchanged between the surface of two epithelia to contribute the functions of the respiratory epithelium and the olfactory receptor cells, respectively. PMID:24200894

  9. NCBI nr-aa BLAST: CBRC-OCUN-01-0528 [SEVENS

    Full Text Available CBRC-OCUN-01-0528 ref|XP_795774.2| PREDICTED: similar to plus agglutinin [Strongylo...centrotus purpuratus] ref|XP_001192339.1| PREDICTED: similar to plus agglutinin [Strongylocentrotus purpuratus] XP_795774.2 0.062 32% ...

  10. Main: 1WGT [RPSD[Archive

    Full Text Available tivum Wheat Germ Agglutinin (Isolectin 3) Lectin (Agglutinin) K.Harata, H.Nagahora, Y.Jigami K.Harata, H.Nagahora, Y.Jigami X-Ray...0026; CHIT_BIND_I_1; 4.|PROSITE; PS50941; CHIT_BIND_I_2; 4. X-Ray Diffraction Len

  11. Cold autoimmune haemolytic anaemia secondary to Epstein Barr virus infection presenting with peripheral gangrene; case report

    Karunarathne Suneth


    Full Text Available Abstract A sixty year old male presented with dark urine, symptomatic anaemia and peripheral gangrene following cold exposure. Investigations revealed that he had haemolysis and serological evidence of recent Epstein Barr virus infection. Although acrocyanosis is commonly associated with cold agglutinin disease, gangrene is a rare complication. Management of secondary cold agglutinin disease is mainly supportive.

  12. Cold autoimmune haemolytic anaemia secondary to Epstein Barr virus infection presenting with peripheral gangrene; case report

    Karunarathne Suneth; Weerasinghe Sajitha; Govindapala Dumitha; Fernando Harshini; Jayaratne Bhaddika


    Abstract A sixty year old male presented with dark urine, symptomatic anaemia and peripheral gangrene following cold exposure. Investigations revealed that he had haemolysis and serological evidence of recent Epstein Barr virus infection. Although acrocyanosis is commonly associated with cold agglutinin disease, gangrene is a rare complication. Management of secondary cold agglutinin disease is mainly supportive.

  13. A lectin histochemical study on carbohydrate moieties of the gonadotropin-like substance in the epithelial cells of Hatschek's pit of Branchiostoma belcheri

    Fang, Y. Q.; Welsch, U.


    The present light microscopic lectin, histochemical study suggests for the first time that the vertebrate gonadotropin-like substance in the basal part of the epithelial cells of Hatschek's pit is a sialic acid-containing glycoprotein. The binding intensity of the epithelial cells in Hatschek's pit to 6 lectins ( Limulus polyphemus agglutinin (LPA), Wheat germ agglutinin (WGA), Helix pomatia agglutinin (HPA), Concanavalin A (Con A), Ulex europaeus agglutinin I (UEA I) and Ricinus communis agglutinin I (RCA I)) indicate that the carbohydrate composition of the gonadotrophic glycoprotein is similar to that of mammals and fish, and that N-acetyl-D-galactosamine, sialic acid, glucosamine, D-mannose and L-fucose are components of the carbohydrate portion.

  14. Genetic labeling of both the axons of transduced, glutamatergic neurons in rat postrhinal cortex and their postsynaptic neurons in other neocortical areas by Herpes Simplex Virus vectors that coexpress an axon-targeted ß-galactosidase and wheat germ agglutinin from a vesicular glutamate transporter-1 promoter

    Zhang, Guo-rong; Cao, Haiyan; Li, Xu; Zhao, Hua; Geller, Alfred I.


    Neuronal circuits comprise the foundation for neuronal physiology and synaptic plasticity, and thus for consequent behaviors and learning, but our knowledge of neocortical circuits is incomplete. Mapping neocortical circuits is a challenging problem because these circuits contain large numbers of neurons, a high density of synapses, and numerous classes and subclasses of neurons that form many different types of synapses. Expression of specific genetic tracers in small numbers of specific sub...

  15. Distribution of lectin-bindings in the testis of the lesser mouse deer, Tragulus javanicus.

    Agungpriyono, S; Kurohmaru, M; Kimura, J; Wahid, A H; Sasaki, M; Kitamura, N; Yamada, J; Fukuta, K; Zuki, A B


    The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d-galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N-acetyl-d-galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N-acetyl-D-glucosamine and sialic acid (wheat germ agglutinin WGA, s-WGA), D-mannose and d-glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), L-fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA-E) sugar residues. In Golgi-, cap-, and acrosome-phase spermatids, lectin-bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s-WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA-E). s-WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA-E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA-E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA-E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin-bindings noted in the testis of lesser mouse deer included the limited distribution of s-WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications. PMID:19245668

  16. A lectin histochemical study on the testis of the babirusa, Babyroussa babyrussa (Suidae).

    Agungpriyono, S; Kurohmaru, M; Prasetyaningtyas, W E; Kaspe, L; Leus, K Y G; Sasaki, M; Kitamura, N; Yamada, J; Macdonald, A A


    The distribution of lectin bindings in the testis of babirusa, Babyrousa babyrussa (Suidae) was studied histochemically using 10 biotinylated lectins, Peanut agglutinin (PNA), Ricinus communis agglutinin (RCA I), Dolichos biflorus agglutinin (DBA), Vicia villosa agglutinin (VVA), Soybean agglutinin (SBA), Wheat germ agglutinin (WGA), Lens culinaris agglutinin (LCA), Pisum sativum agglutinin (PSA), Concanavalin A(Con A) and Ulex europaeus agglutinin (UEA I). Nine of 10 lectins showed a variety of staining patterns in the seminiferous epithelium and interstitial cells. The acrosome of Golgi-, cap- and acrosome-phase spermatids displayed various PNA, RCA I, VVA, SBA and WGA bindings, indicating the presence of glycoconjugates with D-galactose, N-acetyl-D-galactosamine and N-acetyl-D-glucosamine sugar residues respectively. No affinity was detected in the acrosome of late spermatids. LCA, PSA and Con A which have affinity for D-mannose and D-glucose sugar residues were positive in the cytoplasm of spermatids and spermatocytes. DBA was positive only in spermatogonia. In addition to DBA, positive binding in spermatogonia was found for VVA, WGA and Con A, suggesting the distribution of glycoconjugates with N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, D-mannose and D-glucose sugar residues. Sertoli cells were stained intensely with RCA I, WGA and Con A. In Leydig cells, RCA I and Con A were strongly positive, while WGA, LCA and PSA reactions were weak to moderate. The present findings showed that the distribution pattern of lectin binding in the testis of babirusa is somewhat different from that of pig or other mammals reported previously. PMID:17845223

  17. Salmonella enterocolitis

    ... infection if you have: Eaten foods such as turkey, turkey dressing, chicken, or eggs that have not been ... skin. Tests that may be done include: Blood culture Complete blood count with differential Febrile/cold agglutinins ( ...

  18. Cellular carbohydrate components in human, rabbit and rat lacrimal gland. Studies using fluorescein and peroxidase labelled lectins.

    Ahmed, A; Grierson, I


    Orbital lacrimal glands from adult male and female rabbits, rats and humans were examined for the presence of intracellular receptors of four lectins: concanavalin-A agglutinin, lutus tetragonolobus agglutinin, ricinus comunis-60 agglutinin and wheat-germ agglutinin using fluorescein-conjugated lectin and peroxidase labelling methods for fluorescence and electron microscopy, respectively. Lectins were used as specific probes to detect carbohydrate moiety of the lacrimal gland. The pattern of labelling with the lectins suggests that N-acetyl-glucosamine, N-acetyl-D-galactosamine, D-galactose, D-mannose, sialic acid and L-fucose are contained in the lacrimal gland of the three species. The significance of these findings is discussed. PMID:2920911

  19. Use of an Intravascular Warming Catheter during Off-Pump Coronary Artery Bypass Surgery in a Patient with Severe Cold Hemagglutinin Disease.

    Tholpady, Ashok; Bracey, Arthur W; Baker, Kelty R; Reul, Ross M; Chen, Alice J


    Cold hemagglutinin disease with broad thermal amplitude and high titers presents challenges in treating cardiac-surgery patients. Careful planning is needed to prevent the activation of cold agglutinins and the agglutination of red blood cells as the patient's temperature drops during surgery. We describe our approach to mitigating cold agglutinin formation in a 77-year-old man with severe cold hemagglutinin disease who underwent off-pump coronary artery bypass surgery without the use of preoperative plasmapheresis. This experience shows that the use of an intravascular warming catheter can maintain normothermia and prevent the activation and subsequent formation of cold agglutinins. To our knowledge, this is the first reported use of this technique in a patient with cold hemagglutinin disease. The chief feature in this approach is the use of optimal thermal maintenance-rather than the more usual decrease in cold-agglutinin content by means of therapeutic plasma exchange. PMID:27547154

  20. Evaluation of glycophenotype in prostatic neoplasm by chemiluminescent assay

    da Silva, Lúcia Patrícia Bezerra Gomes; Almeida, Sinara Mônica Vitalino; Lima, Luiza Rayanna Amorim; Cavalcanti, Carmelita de Lima Bezerra; Lira, Mariana Montenegro de Melo; da Silva, Maria da Paz Carvalho; Beltrão, Eduardo Isidoro Carneiro; Júnior, Luiz Bezerra de Carvalho


    This work aimed to evaluate the glycophenotype in normal prostate, bening prostatic hyperplasia (BPH) and prostatic adenocarcinoma (PCa) tissues by a chemiluminescent method. Concanavalin A (Con A), Ulex europaeus agglutinin (UEA-I) and Peanut agglutinin (PNA) lectins were conjugated to acridinium ester (lectins-AE). These conjugates remained capable to recognize their specific carbohydrates. Tissue samples were incubated with lectins-AE. The chemiluminescence of the tissue-lectin-AE complex ...

  1. Main: 1K7T [RPSD[Archive

    Full Text Available 1K7T 小麦 Bread Wheat ... Triticum aestivum Agglutinin Isolectin 3 Precursor Triticum Aestivum Molec ... ata M.Muraki, M.Ishimura, K.Harata Interactions Of Wheat -Germ Agglutinin With Glcnacbeta1, 6gal Sequence Bi ... phys.Acta V.1569 10 2002 Hevein-Type Fold SWS:AGI3_WHEAT ,P10969|EMBL; J02961; AAA34257.1; -.|PIR; A28401; A ...

  2. Main: 1K7V [RPSD[Archive

    Full Text Available 1K7V 小麦 Bread Wheat ... Triticum aestivum Agglutinin Isolectin 3 Precursor Triticum Aestivum Molec ... ata M.Muraki, M.Ishimura, K.Harata Interactions Of Wheat -Germ Agglutinin With Glcnacbeta1, 6gal Sequence Bi ... phys.Acta V.1569 10 2002 Hevein-Type Fold SWS:AGI3_WHEAT ,P10969|EMBL; J02961; AAA34257.1; -.|PIR; A28401; A ...

  3. Main: 1K7U [RPSD[Archive

    Full Text Available 1K7U 小麦 Bread Wheat ... Triticum aestivum Agglutinin Isolectin 3 Precursor Triticum Aestivum Molec ... ata M.Muraki, M.Ishimura, K.Harata Interactions Of Wheat -Germ Agglutinin With Glcnacbeta1, 6gal Sequence Bi ... phys.Acta V.1569 10 2002 Hevein-Type Fold SWS:AGI3_WHEAT ,P10969|EMBL; J02961; AAA34257.1; -.|PIR; A28401; A ...

  4. Lectins with Anti-HIV Activity: A Review

    Ouafae Akkouh; Tzi Bun Ng; Senjam Sunil Singh; Cuiming Yin; Xiuli Dan; Yau Sang Chan; Wenliang Pan; Randy Chi Fai Cheung


    Lectins including flowering plant lectins, algal lectins, cyanobacterial lectins, actinomycete lectin, worm lectins, and the nonpeptidic lectin mimics pradimicins and benanomicins, exhibit anti-HIV activity. The anti-HIV plant lectins include Artocarpus heterophyllus (jacalin) lectin, concanavalin A, Galanthus nivalis (snowdrop) agglutinin-related lectins, Musa acuminata (banana) lectin, Myrianthus holstii lectin, Narcissus pseudonarcissus lectin, and Urtica diocia agglutinin. The anti-HIV al...

  5. Lectin histochemistry of palatine glands in the developing rat.

    Hakami, Zaki; Kitaura, Hideki; Honma, Shiho; Wakisaka, Satoshi; Takano-Yamamoto, Teruko


    This study examined the binding pattern of lectins, soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Vicia villosa agglutinin (VVA), Ulex europaeus agglutinin-I (UEA-I), peanut agglutinin (PNA), wheat germ agglutinin (WGA), and succinylated WGA (sucWGA) in the developing rat palatine glands. In adult rats, heterogeneous lectin binding patterns were revealed between the anterior and posterior portions of palatine glands, as DBA, VVA, and WGA were bound more intensely and broadly in the posterior portion. SBA, PNA, and sucWGA showed far less reactivity in the anterior than in the posterior portion. At embryonic day 18 (E18), weak labeling was observed with UEA-I and WGA at the basal membrane of terminal buds, UEA-I and PNA labeled the epithelial cord, and there was no apparent binding for SBA, DBA, VVA, and sucWGA. At E20, after acinar lumenization, all lectins were detected at the acinar cell basal membranes. After birth, all lectins detectably labeled at the mucous cell apical membranes and progressively, with maturation, extended from the apical to basal portions of the cytoplasm. Apparent serous cells were observed around postnatal day 10 (PN10) and bound UEA-I. Lectins reached peak reactivity at PN21 and the binding patterns became identical to those of adults around PN28. PMID:24345684

  6. Diversidade antigênica entre amostras de Arcobacter spp isoladas de suínos no Rio Grande do Sul e presença de anticorpos aglutinantes em amostras de soro de porcas com problemas reprodutivos Antigenic diversity among strains of Arcobacter spp isolated from pigs in Rio Grande do Sul, Brazil and presence of agglutinin titers in serum samples of sows with reproductive problems

    Sérgio José de Oliveira; David Emilio Santos Neves de Barcellos; Sandra Maria Borowski


    O teste de aglutinação microscópica, usando a técnica descrita para o diagnóstico de leptospirose, foi utilizado para verificar a antigenicidade de 47 amostras de Arcobacter cryaerophilus e duas amostras de Arcobacter butzleri isoladas de suínos no Rio Grande do Sul, Brasil, em frente a soros hiperimunes produzidos em coelhos a partir de amostras padrões das bactérias. Verificou-se grande heterogeneidade antigênica e apenas quatro amostras provocaram títulos acima de 1.600 com os anti-soros p...

  7. Flow cytometric analysis of lectin binding to in vitro-cultured Perkinsus marinus surface carbohydrates

    Gauthier, J.D.; Jenkins, J.A.; La Peyre, Jerome F.


    Parasite surface glycoconjugates are frequently involved in cellular recognition and colonization of the host. This study reports on the identification of Perkinsus marinus surface carbohydrates by flow cytometric analyses of fluorescein isothiocyanate-conjugated lectin binding. Lectin-binding specificity was confirmed by sugar inhibition and Kolmogorov-Smirnov statistics. Clear, measurable fluorescence peaks were discriminated, and no parasite autofluorescence was observed. Parasites (GTLA-5 and Perkinsus-1 strains) harvested during log and stationary phases of growth in a protein-free medium reacted strongly with concanavalin A and wheat germ agglutinin, which bind to glucose-mannose and N-acetyl-D-glucosamine (GlcNAc) moieties, respectively. Both P. marinus strains bound with lower intensity to Maclura pomifera agglutinin, Bauhinia purpurea agglutinin, soybean agglutinin (N-acetyl-D-galactosamine-specific lectins), peanut agglutinin (PNA) (terminal galactose specific), and Griffonia simplicifolia II (GlcNAc specific). Only background fluorescence levels were detected with Ulex europaeus agglutinin I (L-fucose specific) and Limulus polyphemus agglutinin (sialic acid specific). The lectin-binding profiles were similar for the 2 strains except for a greater relative binding intensity of PNA for Perkinsus-1 and an overall greater lectin-binding capacity of Perkinsus-1 compared with GTLA-5. Growth stage comparisons revealed increased lectin-binding intensities during stationary phase compared with log phase of growth. This is the first report of the identification of surface glycoconjugates on a Perkinsus spp. by flow cytometry and the first to demonstrate that differential surface sugar expression is growth phase and strain dependent. ?? American Society of Parasitologists 2004.

  8. Different glycosylation of cadherins from human bladder non-malignant and cancer cell lines

    Lityńska Anna


    Full Text Available Abstract Background The aim of the present study was to determine whether stage of invasiveness of bladder cancer cell lines contributes to alterations in glycan pattern of their cadherins. Results Human non-malignant epithelial cell of ureter HCV29, v-raf transfected HCV29 line (BC3726 and transitional cell cancers of urine bladder Hu456 and T24 were grown in cell culture. Equal amounts of protein from each cell extracts were separated by SDS-PAGE electrophoresis and were blotted on an Immobilon P membrane. Cadherins were immunodetected using anti-pan cadherin mAb and lectin blotting assays were performed, in parallel. N-oligosaccharides were analysed by specific reaction with Galanthus nivalis agglutinin (GNA, Sambucus nigra agglutinin (SNA, Maackia amurensis agglutinin (MAA, Datura stramonium agglutinin (DSA, Aleuria aurantia agglutinin (AAA, Phaseolus vulgaris agglutinin (PHA-L and wheat germ agglutinin (WGA. The cadherin from HCV29 cell line possessed bi- and/or 2,4-branched triantennary complex type glycans, some of which were α2,6-sialylated. The cadherin from BC3726 cell line exhibited exclusively high mannose type glycans. Cadherins from Hu456 and T24 cell lines expressed high mannose type glycans as well as β1,6-branched oligosaccharides with poly-N-acetyllactosamine structures and α2,3-linked sialic acid residues. Additionally, the presence of fucose and α2,6-sialic acid residues on the cadherin from T24 cell line was detected. Conclusions These results indicate that N-glycosylation pattern of cadherin from bladder cancer cell line undergoes modification during carcinogenesis.

  9. The typing of Staphylococcus epidermidis by a lectin-binding assay

    Jarløv, J O; Hansen, J E; Rosdahl, V T;


    A new typing method for Staphylococcus epidermidis was developed. Four biotinylated lectins--wheat germ agglutinin (WGA), soy bean agglutinin (SBA), lentil agglutinin (LCA) and Concanavalin A (ConA)--were added to immobilised whole cells of coagulase-negative staphylococci (CNS) in microtitration...... plates. The amount of bound lectin was measured by peroxidase-conjugated avidin followed by a peroxidase reaction. The method was compared to antibiotic-resistance analysis, phage typing, plasmid DNA profiles and slime production. A total of 113 isolates of CNS from 21 patients was investigated and 71...... strains of CNS, including 64 strains of S. epidermidis, were detected if all typing methods were taken into consideration. If only one typing method was used the highest discriminatory power among the S. epidermidis isolates was obtained with the lectin-binding assay which allowed 49 different strains to...

  10. Serine versus threonine glycosylation with α-O-GalNAc: unexpected selectivity in their molecular recognition with lectins.

    Madariaga, David; Martínez-Sáez, Nuria; Somovilla, Víctor J; García-García, Laura; Berbis, M Álvaro; Valero-Gónzalez, Jessika; Martín-Santamaría, Sonsoles; Hurtado-Guerrero, Ramon; Asensio, Juan L; Jiménez-Barbero, Jesús; Avenoza, Alberto; Busto, Jesús H; Corzana, Francisco; Peregrina, Jesús M


    The molecular recognition of several glycopeptides bearing Tn antigen (α-O-GalNAc-Ser or α-O-GalNAc-Thr) in their structure by three lectins with affinity for this determinant has been analysed. The work yields remarkable results in terms of epitope recognition, showing that the underlying amino acid of Tn (serine or threonine) plays a key role in the molecular recognition. In fact, while Soybean agglutinin and Vicia villosa agglutinin lectins prefer Tn-threonine, Helix pomatia agglutinin shows a higher affinity for the glycopeptides carrying Tn-serine. The different conformational behaviour of the two Tn biological entities, the residues of the studied glycopeptides in the close proximity to the Tn antigen and the topology of the binding site of the lectins are at the origin of these differences. PMID:25111627

  11. [Studies on the location of eight lectins in breast carcinoma].

    Zheng, Z; Ji, Z M


    100 cases of breast carcinoma were studied with lectin affinitive histochemistry technology. The result showed that Ricinus comunis agglutinin (RCA1) was located in almost all intraductal carcinomas but one, while the positive rates in the other types were obviously low (P less than 0.05). The positive rate of Ulex europaeus agglutinin-I (UEA1) in well-differentiated types was higher than that in poorly-differentiated ones (P less than 0.05). The location of Peanut agglutinin (PNA), Bandeiraea Simplicifolia (BSL) and UEA1 in breast carcinomas exhibited some regularity and it might be useful in understanding the differentiation of breast carcinomas. No relationship between changes of the eight lectins and metastases in axillary lymph nodes was observed, but the authors considered that PNA-affinitive histochemistry was beneficial to the detection of micrometastases in lymph nodes. PMID:1964401

  12. Characterization of colonic cellular glycoconjugates in colitis and cancer-prone tamarins versus colitis and cancer-resistant primates.

    Moore, R; King, N; Alroy, J


    Differences in colonic secretory glycoconjugates (ie, mucin) between normal and ulcerative colitis-prone patients have been noted. Similar differences may occur in a corresponding primate model, the cotton-top tamarin (CTT), Saguinus oedipus, a New World monkey which suffers from spontaneous chronic colitis and colon cancer. Lectin reagents were used to characterize and compare colonic cell surface, cytoplasmic, and secretory glycoconjugates of 9 clinically healthy cotton-top tamarins, 7 colitis-susceptible, cancer-resistant tamarins (Callithrix jacchus, Saguinus fuscicollis), and 8 colitis and cancer-resistant primates (Aotus trivirgatus, Saimiri sciureus, Macaca fascicularis, and Macaca mulatta). Paraffin-embedded colonic sections were labeled with ten different biotinylated lectins and visualized by the avidin-biotin peroxidase (ABC) method. Significant differences were demonstrated in the pattern of lectin staining between the colitis-resistant and colitis-prone groups of primates. The differences were noted with Griffonia simplicifolia-I (GS-I), Dolichos biflorus agglutinin (DBA), peanut agglutinin (PNA) before and after neuraminidase, Ricinus communis agglutinin-I (RCA-I), soybean agglutinin (SBA), Ulex europaeus agglutinin-I (UEA-I), wheat germ agglutinin (WGA), and succinylated WGA (S-WGA). Significant differences between the CTT and phylogenetically related colitis-prone but cancer-resistant tamarins were demonstrated with SBA, UEA-I, and PNA after desialylation with neuraminidase. These results suggest that differences in colonic cellular glycoconjugates between colitis- and cancer-susceptible species versus colitis-susceptible, cancer-resistant species may be associated with risk of cancer. PMID:3132857

  13. Morphometric Measurements to Quantify the Cerulein Induced Hyperstimulatory Pancreatitis of Rats under the Protective Effect of Lectins

    Jonas, Ludwig; Mikkat, Ulrike; Witte, Anke; Beckmann, Uta; Dölker, Katrin; Weber, Heike; Hahnel, Christian; Kundt, Günther; Nizze, Horst


    In preceding papers we demonstrated an inhibitory effect of wheat germ agglutinin (WGA) and Ulex europaeus agglutinin (UEA) on the cholecystokinin (CCK) binding to the CCK receptor of rat pancreatic cells and also on the CCK induced Ca2+ release and α-amylase secretion in vitro as well as on pancreatic secretion of intact rats in vivo. In the present study we show the same inhibitory effect of both lectins on the cerulein pancreatitis of rats. This acute pancreatitis was induced by supramaxim...

  14. Anopheles gambiae heat shock protein cognate 70B impedes o'nyong-nyong virus replication

    Higgs Stephen; Vanlandingham Dana L; Tsetsarkin Konstantin A; Hong Young S; Sim Cheolho; Collins Frank H


    Abstract Background Phylogenetic and functional analysis was conducted on an Anopheles gambiae gene, ENSANGG00000017398. Based on phylogenetic analysis, this gene belongs to the same lineage as Heat shock protein cognate 70-4 (Hsc70-4) in Drosophila. Accordingly, we propose to name this gene Heat shock protein cognate 70B (HSC70B). We previously reported that expression of HSC70B and other genes including elongation factor-1α (EF-1α) and the agglutinin attachment subunit (agglutinin) were up-...

  15. Characterisation of N-glycans bound to IGFBP-3 in sera from healthy adults.

    Masnikosa, Romana; Baricević, Ivona; Lagundzin, Dragana; Nedić, Olgica


    Human IGFBP-3 contains three potential N-linked glycosylation sites. Published data concerning the type and saccharide composition of the N-glycans is scarce. The aim of this study was to characterise N-glycans covalently attached to IGFBP-3 from sera of healthy adults (men and women). In order to do that a panel of eight lectins covering broad saccharide specificity was used: agarose-immobilised SNA (Sambucus nigra agglutinin), Con A (lectin from Canavalia ensiformis), RCA I (Ricinus communis agglutinin I), PHA-E (Phaseolus vulgaris erythroagglutinin), PHA-L (P. vulgaris leukoagglutinin), succinylated WGA (wheat germ agglutinin), ECL (Erythrina cristagalli lectin) and UEA (Ulex europaeus agglutinin). IGFBP-3 interacted with SNA, Con A, RCA I, PHA-E and, to a much lesser extent, with PHA-L. These results indicate that human IGFBP-3 bears mostly biantennary complex type N-glycans with a very high content of alpha-2,6-linked Sia at their termini. Hybrid type and high-mannose type N-glycans are present, as well as a bisecting GlcNAc residue, which may be core fucosylated. N-glycosylation of IGFBP-3 follows the N-glycosylation pattern of major serum proteins. This study represents a ground for the future research of glycosylation pattern of IGFBP-3 from the circulation of men and women diagnosed with different illnesses. PMID:19800385

  16. Deep trophoblast invasion and spiral artery remodelling in the placental bed of the lowland gorilla

    Pijnenborg, R; Vercruysse, L; Carter, Anthony Michael


    chimpanzee, we postulated the occurrence of deep invasion in gorilla pregnancy. Tissues were processed for histology (PAS, orcein), lectin staining (Ulex europaeus agglutinin 1) and immunohistochemistry (cytokeratin 7/17, α-actin). A specimen of young but undetermined gestational age included deep placental...

  17. Deep trophoblast invasion and spiral artery remodelling in the placental bed of the chimpanzee

    Pijnenborg, R; Vercruysse, L; Carter, Anthony Michael


    -actin, as well as Ulex europaeus agglutinin 1 (UEA1) lectin staining, in this archival material. In both specimens interstitial trophoblast invasion had occurred in both decidua and myometrium. Because of a lack of published data on fetal growth for this species, fetal sizes (7cm and 13cm) could not be...

  18. Haemangioblastoma, Histological and immunohistological study of an enigmatic cerebellar tumour

    Cruz-Sánchez, F. F.; Rossi, M L; Rodríguez-Prados, S.; Nakamura, N; Hughes, J T; Coakham, H. B.


    Paraffin-embedded blocks of 36 cerebellar haemangioblastomas were reacted with a panel of antibodies including glial fibrillary acidic protein, vimentin, epithelial membrane antigen, cytokeratin, Factor VIII, a neuroendocrine marker and with Ulex europaeus. agglutinin The main histological features, apart from the characteristic large abnormal vessels, were a prominent reticulin network, a cystic architecture and cellular and nuclear polymorphism. Two cell type...

  19. A comparative analysis of the attachment of Leptospira interrogans and L. borgpetersenii to mammalian cells.

    Andrade, Gabrielle I; Brown, Paul D


    Leptospirosis, the world's most ubiquitous zoonosis, is caused by pathogenic Leptospira. As microbe-host interactions are specific in pathogenesis, it is likely that there are several molecules mediating the attachment of the Leptospira to mammalian cells. In this study, we analysed the attachment of Leptospira interrogans serovar Portlandvere and Leptospira borgpetersenii serovar Jules to untreated HEp-2 cells or HEp-2 cells treated with the various enzymes, lectins or sugars and to integrins αVβ3 and α5β1, relative to control wells. We found that both serovars bound equally well to HEp-2 cells; however, serovar Jules showed a higher level of attachment to integrins. Both serovars showed an increase in attachment to HEp-2 cells coated with lectins peanut agglutinin, Ulex europaeus agglutinin, soybean agglutinin and Erythrina cristagalli agglutinin (p < 0.05); in the case of Concanavalin A, Jules showed an increase, while Portlandvere showed a significant decrease in attachment. Trypsinizing monolayers resulted in a decrease in attachment for both serovars, while when chondroitinase, neuraminidase and heparinase were used an increase in attachment was recorded. Leptospires coated with sugars showed a decrease in attachment. These results show that serovar Jules' general greater affinity for the mediators examined may suggest a greater potential for virulence over serovar Portlandvere. PMID:22409511

  20. The diagnosis of unilateral testicular obstruction in subfertile males.

    Hendry, W F; Parslow, J M; Stedronska, J; Wallace, D M


    Thirty-two subfertile males with sperms in the ejaculate and unilateral testicular obstruction are reported: the diagnosis was established by exploration of scrotum in 26, clinically in 2, 3 had had previous partially successful epididymovasostomies, and 1 had had an epispadias repair. The past medical history gave relevant information in 27 (84%), and useful findings were made on clinical examination in a further 3 cases. Fifteen patients had sperm counts over 20 million per ml, and 15 were less than 10 million per ml. Twenty-six (81%) had serum antisperm antibodies detected by tray agglutination test (TAT), 21 (81%) of whom had evidence of head-to-head (HH) agglutinins in pure or mixed form. Comparison with 162 vasectomised males and 160 naturally infertile males with antisperm antibodies showed that 55% of the former and 24% of the latter had HH agglutinins on TAT, differences that were highly significant on statistical analysis. Evidence of obstruction was found in 14 (37%) of 38 naturally infertile males with antisperm antibodies and HH or mixed agglutination, but only in 12 (10%) of 122 with TT agglutinins: this difference was also highly significant. Clinical history, physical examination and serum antisperm antibodies, especially if HH agglutinins are present, can suggest the possibility of unilateral testicular obstruction, but confirmation of the diagnosis requires exploration of scrotum. PMID:7150940

  1. Early events of secretory granule formation in the rat parotid acinar cell under the influence of isoproterenol. An ultrastructural and lectin cytochemical study

    F D’Amico


    Full Text Available The events involved in the maturation process of acinar secretory granules of rat parotid gland were investigated ultrastructurally and cytochemically by using a battery of four lectins [Triticum vulgaris agglutinin (WGA, Ulex europaeus agglutinin I (UEA-I, Glycine max agglutinin (SBA, Arachys hypogaea agglutinin (PNA]. In order to facilitate the study, parotid glands were chronically stimulated with isoproterenol to induce secretion. Specimens were embedded in the Lowicryl K4M resin. The trans-Golgi network (TGN derived secretory granules, which we refer to as immature secretory granules, were found to be intermediate structures in the biogenesis process of the secretory granules in the rat parotid acinar cell. These early structures do not seem to be the immediate precursor of the mature secretory granules: in fact, a subsequent interaction process between these early immature granule forms and TGN elements seems to occur, leading, finally, to the mature granules. These findings could explain the origin of the polymorphic subpopulations of the secretory granules in the normal acinar cells of the rat parotid gland. The lectin staining patterns were characteristic of each lectin. Immature and mature secretory gran- ules were labelled with WGA, SBA, PNA, and lightly with UEA-I. Cis and intermediate cisternae of the Golgi apparatus were labelled with WGA, and trans cisternae with WGA and SBA.

  2. Histological and lectin histochemical studies on the main and accessory olfactory bulbs in the Japanese striped snake, Elaphe quadrivirgata.

    Kondoh, Daisuke; Wada, Akimi; Endo, Daisuke; Nakamuta, Nobuaki; Taniguchi, Kazuyuki


    The main and accessory olfactory bulbs were examined by histological methods and lectin histochemistry in the Japanese striped snake. As the results, the histological properties are similar between the main olfactory bulb and the accessory olfactory bulb. In lectin histochemistry, 21 lectins used in this study showed similar binding patterns in the main olfactory bulb and the accessory olfactory bulb. In detail, 15 lectins stained these olfactory bulbs with similar manner, and 6 lectins did not stain them at all. Two lectins, Lycopersicon esculentum lectin (LEL) and Solanum tuberosum lectin (STL), stained the nerve and glomerular layers and did not stain any other layers in both olfactory bulbs. Four lectins, Soybean agglutinin (SBA), Vicia villosa agglutinin (VVA), Peanut agglutinin (PNA) and Phaseolus vulgaris agglutinin-L (PHA-L) stained the nerve and glomerular layers more intensely than other layers in both olfactory bulbs. In addition, VVA showed the dot-like stainings in the glomeruli of both olfactory bulbs. These findings suggest that the degree of development and the properties of glycoconjugates are similar between the main olfactory bulb and the accessory olfactory bulb in the Japanese striped snake. PMID:23257605

  3. Ultrastructural and histochemical properties of the olfactory system in the japanese jungle crow, Corvus macrorhynchos.

    Kondoh, Daisuke; Nashimoto, Mai; Kanayama, Shunsaku; Nakamuta, Nobuaki; Taniguchi, Kazuyuki


    Although it has been commonly believed that birds are more dependent on the vision and audition than the olfaction, recent studies indicate that the olfaction of birds is related to the reproductive, homing, and predatory behaviors. In an attempt to reveal the dependence on the olfactory system in crows, we examined the olfactory system of the Japanese jungle crow (Corvus macrorhynchos) by histological, ultrastructural, and lectin histochemical methods. The olfactory epithelium (OE) of the crow occupied remarkably a small area of the nasal cavity (NC) and had the histological and ultrastructural features like other birds. The olfactory bulb (OB) of the crow was remarkably small and did not possess the olfactory ventricle. The left and right halves of the OB were fused in many cases. In the lectin histochemistry, soybean agglutinin (SBA) and Vicia villosa agglutinin (VVA) stained a small number of the receptor cells (RCs) in the OE and the olfactory nerve layer (ONL) and glomerular layer (GL) on the dorsocaudal region of the OB. Phaseolus vulgaris agglutinin-E (PHA-E) stained several RCs in the OE and the ONL and GL on the ventral region of the OB. These results suggest that 1) the crow has less-developed olfactory system than other birds, and 2) the dedicated olfactory receptor cells project their axons to the specific regions of the OB in the crow. PMID:21478653

  4. Primary breast cancer tumours contain high amounts of IgA1 immunoglobulin

    Welinder, Charlotte; Baldetorp, Bo; Blixt, Klas Ola;


    The Tn antigen (GalNAc alpha-O-Ser/Thr) as defined by the binding of the lectin, helix pomatia agglutinin (HPA) or anti-Tn monoclonal antibodies, is known to be exposed in a majority of cancers, and it has also been shown to correlate positively with the metastatic capacity in breast carcinoma. T...

  5. Quantitative approach to lectin-based glycoprofiling of thymic tissues in the control- and the dexamethasone-treated mice.

    Balcan, Erdal


    Dexamethasone (DEX) is the most commonly used synthetic glucocorticoid in treatment of various inflammatory conditions. Here we focused on evaluating the effect of DEX on apoptosis and glycan profile in the mouse thymic tissues. Histological examinations revealed that the DEX treatment cause severe alterations in thymus, such as disruption of thymic capsule, impaired epithelial cell-thymocyte contacts, cellular loss and increased apoptosis. The identification of thymic glycans in the control- and the DEX-treated mice was carried out by using a panel of five plant lectins, Maackia amurensis agglutinin (MAA), peanut agglutinin (PNA), Sambucus nigra agglutinin (SNA), Concanavalin A (ConA) and wheat germ agglutinin (WGA). Lectin histochemistry results showed that glycosylation pattern of thymus changes upon DEX treatment. For further detailed quantitative analyses of the binding intensities for each lectin, histochemical data were scored as high positive (HP), mild positive (MP) and low positive (LP) and differences among signaling densities were investigated. The staining patterns of thymic regions observed with lectin histochemistry suggest that DEX can affect the thymic glycan profile as well as thymocyte apoptosis. These results are consistent with the opinion that not only sialic acid, but also other sugar motifs may be responsible for thymocyte development. PMID:27067421

  6. Gastrointestinal nematodes and anthelmintic resistance in Danish goat herds

    Holm, Signe A.; Sørensen, Camilla; Thamsborg, Stig M.;


    stained with peanut agglutinin (PNA) for specific detection of Haemonchus contortus. Strongyle eggs were detected with an individual prevalence of 69%, including Nematodirus battus (3.6%) and other Nematodirus species (15.0%). Eimeria spp. were observed in 99.6% of the kids. H. contortus was found in 11...

  7. Partial Purification of the 5-Hydroxytryptamine-Reuptake System from Human Blood Platelets Using a Citalopram-Derived Affinity Resin

    Biessen, E.A.L.; Horn, A.S.; Robillard, G.T.


    This paper describes a procedure for the synthesis and application of a citalopram-derived affinity resin in purifying the 5HT-reuptake system from human blood platelets. A two-step scheme has been developed for partial purification, based on wheat germ agglutinin-lectin (WGA) affinity and citalopra

  8. NCBI nr-aa BLAST: CBRC-MDOM-02-0035 [SEVENS

    Full Text Available protein, putative; cell-surface glycoprotein, putative [Candida dubliniensis CD36] emb|CAX43113.1| ALS protein, putative; agglutinin-like protein, putative; cell-surface glycoprotein, putative [Candida dubliniensis CD36] XP_002419518.1 5e-46 22% ...

  9. A morphological study of the vomeronasal organ and the accessory olfactory bulb in the Korean roe deer, Capreolus pygargus.

    Park, Changnam; Ahn, Meejung; Lee, Jae-Yuk; Lee, Sang; Yun, Youngmin; Lim, Yoon-Kyu; Taniguchi, Kazumi; Shin, Taekyun


    The vomeronasal organ (VNO) and accessory olfactory bulb (AOB) of the Korean roe deer (Capreolus pygargus) were studied histologically to evaluate their morphological characteristics. Grossly, the VNO, encased by cartilage, has a paired tubular structure with a caudal blind end and a rostral connection through incisive ducts on the hard palate. In the VNO, the vomeronasal sensory epithelium (VSE) consists of galectin-3-positive supporting cells, protein gene product (PGP) 9.5-positive receptor cells, and basal cells. The vomeronasal respiratory epithelium (VRE) consists of a pseudostratified epithelium. The AOB strata included a vomeronasal nerve layer (VNL), a glomerular layer (GL), a mitral/tufted cell layer, and a granular cell layer. All lectins used in this study, including Bandeiraea simplicifolia agglutinin isolectin B4 (BSI-B4), soybean agglutinin (SBA), Ulex europaeus agglutinin I (UEA-I), and Triticum vulgaris wheat germ agglutinin (WGA), labeled the VSE with varying intensity. In the AOB, both the VNL and the GL reacted with BSI-B4, SBA, and WGA with varying intensity, but not with UEA-I. This is the first morphological study of the VNO and AOB of the Korean roe deer, which are similar to those of goats. PMID:24055195

  10. Bacterial, fungal, and algal lectins: combatants in tug of war against HIV.

    Feizi, Ten; Liu, Yan; Palma, Angelina S


    High-resolution X-ray crystallography and NMR studies by Koharudin and Gronenborn in this issue provide new information on the mode of N-glycan recognition by a cyanobacterial agglutinin, with anti-HIV activity pointing to the pentamannosyl core as a novel target for therapeutic intervention. PMID:21827940

  11. Main: 2SBA [RPSD[Archive

    Full Text Available A.Dessen, D.Gupta, S.Sabesan, C.F.Brewer, J.C.Sacchettini X-Ray Crystal Structure Of The Soybean Agglutinin ...; PS00307; LECTIN_LEGUME_BETA; 1. X-Ray Diffraction, Resolution: 2.60 Angstrom, R-Factor:: 0.208, R-Free:: N

  12. Comparison of the binding properties of the mushroom Marasmius oreades lectin and Griffonia simplicifolia I-B isolectin to alphagalactosyl carbohydrate antigens in the surface phase

    Kirkeby, Svend; Winter, Harry C; Goldstein, Irwin J


    The binding of two alpha-galactophilic lectins, Marasmius oreades agglutinin (MOA), and Griffonia simplicifolia I isolectin B(4) (GS I-B(4)) to neoglycoproteins and natural glycoproteins were compared in a surface phase assay. Neoglycoproteins carrying various alpha-galactosylated glycans and lam...

  13. Residu Gula Glikokonjugat pada Lambung Depan Kerbau Rawa (Bubalus bubalis Kalimantan Selatan (SUGAR RESIDU OF GLYCOCONJUGATES IN FORESTOMACH OF SOUTH KALIMANTAN SWAMP BUFFALO (BUBALUS BUBALIS

    Anni Nurliani


    Full Text Available The ability of swamp buffaloes to adapt with swamp environment was suggested to be supported bytheir digestive system efficiency. The research was done to obtain scientific explanation about digestiveefficiency of swamp buffalo by identification on kinds and distribution of glycoconjugates in swamp buffaloforestomach. Six male swamp buffaloes aged more than 2.5 year old and had body weight between 300-400kg were used in this study. Samples were obtained from Regency of Banjar slaughter house, SouthKalimantan. Every parts of the forestomach included rumen, reticulum, and omasum was taken andprocessed for microscopic observation with hematoxyline eosin (HE and alcian blue-periodic acid schiff(AB-PAS stainings. Sugar residues of glycoconjugates were localized with lectin histochemistry wheatgerm agglutinin (WGA, ulex europaeus agglutinin (UEA, ricinus communis agglutinin (RCA, concanavalinagglutinin (Con A, and soybean agglutinin (SBA. Every part of swamp buffalo forestomach had kinds ofspecific glycoconjugates with special distribution pattern which were different with other ruminant, andwere suitable for their functions in that part. The existence of D mannose/D glucose glycoconjugates thatwas dominant in forestomach estimated that had important role in supporting fermentative digestionfunction in swamp buffalo, through its function as receptor bacteria attachment. This is suggested as aspecial characteristic in digestive system of swamp buffalo which causes high digestive efficiency inswamp buffalo.

  14. Lectin binding profiles of SSEA-4 enriched, pluripotent human embryonic stem cell surfaces

    Shin Soojung


    Full Text Available Abstract Background Pluripotent human embryonic stem cells (hESCs have the potential to form every cell type in the body. These cells must be appropriately characterized prior to differentiation studies or when defining characteristics of the pluripotent state. Some developmentally regulated cell surface antigens identified by monoclonal antibodies in a variety of species and stem cell types have proven to be side chains of membrane glycolipids and glycoproteins. Therefore, to examine hESC surfaces for other potential pluripotent markers, we used a panel of 14 lectins, which were chosen based on their specificity for a variety of carbohydrates and carbohydrate linkages, along with stage specific embryonic antigen-4 (SSEA-4, to determine binding quantitation by flow cytometry and binding localization in adherent colonies by immunocytochemistry. Results Enriching cells for SSEA-4 expression increased the percentage of SSEA-4 positive cells to 98–99%. Using enriched high SSEA-4-expressing hESCs, we then analyzed the binding percentages of selected lectins and found a large variation in binding percentages ranging from 4% to 99% binding. Lycopersicon (tomatoesculetum lectin (TL, Ricinus communis agglutinin (RCA, and Concanavalin A (Con A bound to SSEA-4 positive regions of hESCs and with similar binding percentages as SSEA-4. In contrast, we found Dolichos biflorus agglutinin (DBA and Lotus tetragonolobus lectin (LTL did not bind to hESCs while Phaseolus vulgaris leuco-agglutinin (PHA-L, Vicia villosa agglutinin (VVA, Ulex europaeus agglutinin (UEA, Phaseolus vulgaris erythro-agglutinin (PHA-E, and Maackia amurensis agglutinin (MAA bound partially to hESCs. These binding percentages correlated well with immunocytochemistry results. Conclusion Our results provide information about types of carbohydrates and carbohydrate linkages found on pluripotent hESC surfaces. We propose that TL, RCA and Con A may be used as markers that are associated with the

  15. The Glycoprofile Patterns of Endothelial Cells in Usual Interstitial Pneumonia

    A Barkhordari


    Full Text Available [THIS ARTICLE HAS BEEN RETRACTED FOR DUPLICATE PUBLICATION] Background: The pathological classification of cryptogenic fibrosing alveolitis has been a matter of debate and controversy for histopathologists.Objective: To identify and specify the glycotypes of capillary endothelial cells in usual interstitial pneumonia (UIP compared to those found in normal tissue.Methods: Sections of formalin-fixed, paraffin-embedded blocks from 16 cases of UIP were studied by lectin histochemistry with a panel of 27 biotinylated lectins and an avidin-peroxidase revealing system.Results: High expression of several classes of glycan was seen de novo in capillary endothelial cells from patients with UIP including small complex and bi/tri-antennary bisected complex N-linked sequences bolund by Concanavalin A and erythro-phytohemagglutinin, respectively, GalNAca1 residues bound by Helix pomatia and Maclura pomifera agglutinins, and L-fucosylated derivatives of type II glycan chains recognized by Ulex europaeus agglutinin-I. Glycans bound by agglutinins from Lycopersicon esculentum (β1,4GlcNAc and Wisteria floribunda (GalNAc as well as GlcNAc oligomers bound by Phytolacca americana and succinylated Wheat Germ agglutinin were also seen in the capillary endothelial cells of UIP. In contrast, L-fucosylated derivatives of type I glycan chains were absent in cells from cases of UIP when Anguilla anguilla agglutinin was applied, unlike the situation in normal tissue.Conclusion: These results may indicate existence of two distinct populations of endothelial cell in UIP with markedly different patterns of glycosylation, reflecting a pattern of differentiation and angiogenesis, which is not detectable morphologically.

  16. Seroepidemiology of Toxoplasma gondii in dogs in Trinidad and Tobago.

    Ali, C N; Harris, J A; Watkins, J D; Adesiyun, A A


    A cross-sectional study was conducted to determine the seroprevalence of Toxoplasma gondii agglutinins and to investigate the relationship between various risk factors and occurrence of toxoplasmosis in dogs in Trinidad. Of a total 250 dogs, comprising domestic, hunting and stray dogs, 80 (32.0%) were positive for T. gondii agglutinins at a titre of > or =1:32 using a latex agglutination test. Stray dogs (60.5%) had statistically significantly higher (P2-3 years age group compared with other age groups. Dogs that consumed home-cooked foods had a seroprevalence of 32.9% compared with those fed commercial dog foods (17.2%) and dogs fed both home-cooked and commercial foods (21.0%). However, the difference was not statistically significant (P>0.05; chi(2)). The rather high prevalence of toxoplasmosis in stray dogs is a good indication of the extent of the infection in the environment. PMID:12719132

  17. Physico-chemical characterization of bacterial and hemagglutinins from the serum of the mud crab Scylla serrata

    SS Jayaraj


    Full Text Available A naturally occurring hemagglutinin (HA, with activity against bacteria and yeast cells were detected in the serum of Scylla serrata using mammalian erythrocytes (RBC, various bacteria and yeast as indicator cells. The serum gave highest HA titer with rabbit RBC, tripsinized yeast and Vibrio fluvialis. An analysis of the physico-chemical properties of the HA showed it to be specifically dependent on the presence of Ca2+ for its activity, stable between pH 7 to 9 and showed thermal stability between 10 to 30 °C. Further studies demonstrated that the HA is precipitable by ammonium sulphate and TCA. HA- inhibition assays performed with carbohydrates revealed that the serum HA was specific for non-reducing terminal glucose with 1-2 glucosidic linkages. Thus this agglutinin appears to be unique among all the known crustacean agglutinins.

  18. The immunological properties of Brucella ribosomal preparations.

    Corbel, M J


    Ribosomes were isolated from Brucella abortus strains 19 and 45/20 by disruption of the cells followed by differential ultracentrifugation. The ribosome preparations contained 2-3 components reacting in immunodiffusion tests but were free of detectable lipopolysaccharide-protein agglutinogen. They crossreacted with antisera to Br. abortus, Br. melitensis, Br. suis and Br. ovis and elicited intradermal delayed hypersensitivity reactions in animals infected with Br. abortus, Br. melitensis or Br. suis. The ribosomes were antigenic in rabbits, guinea pigs and mice. Those from Br. abortus S19 induced agglutinins reaction with smooth brucella strains whereas those from Br. abortus 45/20 induced agglutinins reacting with rough brucella strains. Cattle vaccinated with S19 or 45/20 vaccines or infected with Br. abortus developed pricipitins to ribosomal components at an early stage in the immune response. PMID:816681


    Danilo Cutrim Bezerra


    Full Text Available With the objective of searching the presence of antileptospiras agglutinin in sanguine serum of donkeys used in animal traction vehicles and of the wagon’s conductors in the city of São Luis, MA, and of identifying serovars of higher occurrence, 60 serum samples of donkeys and 60 of conductors were tested regarding 30 serovars of Leptospira interrogans by using the microscopic seroagglutination test. Of the 60 blood serum samples of donkeys, 85% showed seropositivity, reacting to 21 serovars out of the 30 tested. Copenhageni, Pyrogenes, Autumnalis and Icterohaemorrhagiae were the most frequent ones, in decreasing sequence. Regarding the serum of the conductors, 38.34% presented seropositivity, reacting to 12 serovars of Leptospira interrogans and the most frequenty ones were Copenhageni, Pyrogenes, Icterohaemorrhagiae and Autumnalis. These results indicate a high incidence of anti-leptospira agglutinins in donkeys and humans, being serovars copenhageni and pyrogenes the most prevalent.

  20. AcEST: DK948418 [AcEST

    Full Text Available or domain OS=St... 46 0.002 tr|A2DMT1|A2DMT1_TRIVA A-agglutinin attachment subunit, putative.....ESD 1725 >tr|A2DMT1|A2DMT1_TRIVA A-agglutinin attachment subunit, putative OS=Trichomonas vaginalis G3 GN=TV....3 E-value 2.0e-05 Report BLASTX 2.2.19 [Nov-02-2008] Reference: Altschul, Stephe... adhesin for platelets OS=Staphy... 46 2e-04 sp|Q99QY4|SRAP_STAAM Serine-rich adhesin for platelets... OS=Staphy... 46 2e-04 sp|Q8NUJ3|SRAP_STAAW Serine-rich adhesin for platelets OS=Staphy... 45 5e-04 sp|Q

  1. AcEST: DK961080 [AcEST

    Full Text Available 6 2.0 tr|A2DMT1|A2DMT1_TRIVA A-agglutinin attachment subunit, putative... 35 3.5 tr|Q4SYS6|Q4SYS6_TETNG Chromosome undet...080 Tissue type prothallia with plantlets Developmental stage gametophytes with s... SYD ++P SN Y QT +P+Q Sbjct: 307 LTKFSVISDSTCFSSSK-FTTKSYDIESPFKQSNNYVNQTPQPSQ 350 >tr|A2DMT1|A2DMT1_TRIVA A-agglutinin attachme... 2.2.19 [Nov-02-2008] Reference: Altschul, Stephen F., Thomas L. Madden, Alejandro A. Schaffer, Jinghui Zhan...omonas vaginalis G3 Align length 97 Score (bit) 38.1 E-value 0.53 Report BLASTX 2.2.19 [Nov-02-2008] Reference: Altsch

  2. Increased fucosyl glycoconjugate by Mycoplasma hyopneumoniae enhances adherences of Pasteurella multocida type A in the ciliated epithelial cells of the respiratory tract

    Park, Changhoon; Jeong, Jiwoon; KANG, Ikjae; Choi, Kyuhyung; Park, Su-Jin; Chae, Chanhee


    Background The objective of this study was to elucidate the pathogenic mechanisms of how Mycoplasma hyopneumoniae enhances secondary Pasteurella multocida type A infection which leads to porcine enzootic pneumonia in infected pigs. Sixteen pigs were experimentally infected with M. hyopneumoniae and then euthanized at 7, 14, 21 and 28 days post inoculation. In situ hybridization for M. hyopneumoniae DNA and Ulex europaeus agglutinin-I (UEA-I) lectin histochemistry for fucosyl glycoconjugate, w...

  3. Identification of lectin-binding proteins in Chlamydia species.

    Swanson, A F; Kuo, C. C.


    Lectin-binding proteins of chlamydiae were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. All three Chlamydia species tested expressed two proteins when whole-elementary-body lysates were reacted with the biotinylated lectin Dolichos biflorus agglutinin. The protein with a molecular mass of 18 kilodaltons (kDa) responded strongly compared with a higher-molecular-mass protein that varied from 27 to 32 kDa with each chlamydia strain tested. Among six l...

  4. Glycoconjugates Distribution during Developing Mouse Spinal Cord Motor Organizers

    Vojoudi, Elham; Ebrahimi, Vahid; Ebrahimzadeh-Bideskan, Alireza; Fazel, Alireza


    Background: The aim of this research was to study the distribution and changes of glycoconjugates particularly their terminal sugars by using lectin histochemistry during mouse spinal cord development. Methods: Formalin-fixed sections of mouse embryo (10-16 fetal days) were processed for lectin histochemical method. In this study, two groups of horseradish peroxidase -labeled specific lectins were used: N-acetylgalactosamine, including Dolichos biflorus, Wisteria floribunda agglutinin (WFA), ...

  5. Endothelial progenitor cells regenerate infracted myocardium with neovascularisation development ☆

    M.T. Abd El Aziz; Abd El Nabi, E.A.; Abd El Hamid, M.; D. Sabry; Atta, H.M.; L.A. Rahed; A. Shamaa; Mahfouz, S.; Taha, F.M.; S. Elrefaay; Gharib, D.M.; Elsetohy, Khaled A


    We achieved possibility of isolation, characterization human umbilical cord blood endothelial progenitor cells (EPCs), examination potency of EPCs to form new blood vessels and differentiation into cardiomyoctes in canines with acute myocardial infarction (AMI). EPCs were separated and cultured from umbilical cord blood. Their phenotypes were confirmed by uptake of double stains dioctadecyl tetramethylindocarbocyanine-labeled acetylated LDL and FITC-labeled Ulex europaeus agglutinin 1 (DILDL-...

  6. M-Cell Targeting of Whole Killed Bacteria Induces Protective Immunity against Gastrointestinal Pathogens▿

    Chionh, Yok-Teng; Wee, Janet L. K.; Every, Alison L.; Ng, Garrett Z.; Sutton, Philip


    As the majority of human pathogens infect via a mucosal surface, delivery of killed vaccines by mucosal routes could potentially improve protection against many such organisms. Our ability to develop effective killed mucosal vaccines is inhibited by a lack of adjuvants that are safe and effective in humans. The Ulex europaeus agglutinin I (UEA-I) lectin specifically binds M cells lining the murine gastrointestinal tract. We explored the potential for M-cell-targeted vaccination of whole, kill...

  7. Biochimical and Structural caracterization of a lectin from Platypodium elegans Vogel seeds

    Leite, Raquel,


    Lectin activity with specificity for mannose and glucose has been detected in the seed of Platypodium elegans, a legume from the Dalbergiae tribe. The gene of the lectin PELa has been cloned and the resulting 261 amino acid protein belongs to the legume lectin family with similarity with Pterocarpus angolensis agglutinin (PAL) from the same tribe. The recombinant lectin has been expressed in E. coli and refolded from inclusion bodies. Analysis of specificity by Glycan Array evidenced a very u...

  8. Diagnosis of myocardial infarction based on lectin-induced erythrocyte agglutination: a feasibility study

    Bocsi, József; Nieschke, Kathleen; Mittag, Anja; Reichert, Thomas; Laffers, Wiebke; Marecka, Monika; Pierzchalski, Arkadiusz; Piltz, Joachim; Esche, Hans-Jürgen; Wolf, Günther; Dähnert, Ingo; Baumgartner, Adolf; Tarnok, Attila


    Myocardial infarction (MI) is an acute life-threatening disease with a high incidence worldwide. Aim of this study was to test lectin-carbohydrate binding-induced red blood cell (RBC) agglutination as an innovative tool for fast, precise and cost effective diagnosis of MI. Five lectins (Ricinus communis agglutinin (RCA), Phaseolus vulgaris erythroagglutinin (PHA), Datura stramonium agglutinin (DSA), Artocarpus agglutinin (ArA), Triticum agglutinin (TA)) were tested for ability to differentiate between agglutination characteristics in patients with MI (n = 101) or angina pectoris without MI (AP) (n = 34) and healthy volunteers (HV) as control (n =68) . RBC agglutination was analyzed by light absorbance of a stirred RBC suspension in the green to red light spectrum in an agglutimeter (amtec, Leipzig, Germany) for 15 min after lectin addition. Mean cell count in aggregates was estimated from light absorbance by a mathematical model. Each lectin induced RBC agglutination. RCA led to the strongest RBC agglutination (~500 RBCs/aggregate), while the others induced substantially slower agglutination and lead to smaller aggregate sizes (5-150 RBCs/aggregate). For all analyzed lectins the lectin-induced RBC agglutination of MI or AP patients was generally higher than for HV. However, only PHA induced agglutination that clearly distinguished MI from HV. Variance analysis showed that aggregate size after 15 min. agglutination induced by PHA was significantly higher in the MI group (143 RBCs/ aggregate) than in the HV (29 RBC-s/aggregate, p = 0.000). We hypothesize that pathological changes during MI induce modification of the carbohydrate composition on the RBC membrane and thus modify RBC agglutination. Occurrence of carbohydrate-lectin binding sites on RBC membranes provides evidence about MI. Due to significant difference in the rate of agglutination between MI > HV the differentiation between these groups is possible based on PHA-induced RBC-agglutination. This novel assay

  9. Specificity of lectin-immobilized fluorescent nanospheres for colorectal tumors in a mouse model which more resembles the clinical disease

    Kitamura, Tokio; Sakuma, Shinji; Shimosato, Moe; Higashino, Haruki; Masaoka, Yoshie; Kataoka, Makoto; Yamashita, Shinji; Hiwatari, Ken-Ichiro; Kumagai, Hironori; Morimoto, Naoki; Koike, Seiji; Tobita, Etsuo; Hoffman, Robert M.; John C Gore; Pham, Wellington


    We are investigating an imaging agent that enables real-time and accurate diagnosis of early colorectal cancer at the intestinal mucosa by colonoscopy. The imaging agent is peanut agglutinin-immobilized polystyrene nanospheres with surface poly(N-vinylacetamide) chains encapsulating coumarin 6. Intracolonically-administered lectin-immobilized fluorescent nanospheres detect tumor-derived changes through molecular recognition of lectin for the terminal sugar of cancer-specific antigens on the m...

  10. Histological and Lectin Histochemical Studies on the Olfactory and Respiratory Mucosae of the Sheep

    Ibrahim, Dalia; NAKAMUTA, Nobuaki; TANIGUCHI, Kazumi; Yamamoto, Yoshio; TANIGUCHI, Kazuyuki


    ABSTRACT The olfactory and respiratory mucosae of the Corriedale sheep were examined using lectin histochemistry in order to clarify the histochemical and glycohistochemical differences between these two tissues. The olfactory epithelium was stained with 13 lectins out of 21 lectins examined, while the respiratory epithelium was positive to 16 lectins. The free border of both of the olfactory and respiratory epithelia was stained with 12 lectins: Wheat germ agglutinin (WGA), succinylated-whea...

  11. The A0 blood group genotype modifies the jejunal glycomic binding pattern profile of piglets early associated with a simple or complex microbiota.

    Priori, D; Colombo, M; Koopmans, S-J; Jansman, A J M; van der Meulen, J; Trevisi, P; Bosi, P


    The intestinal epithelium glycocalyx sugar motif is an important determinant of the bacterial-host interaction and may be affected in pigs by gut microbiota and by blood group genotype. The aim was to study the effect of intestinal association with different microbiota and A0 blood group genotypes on the expressed glycomic pattern in the small intestine. Twelve caesarean-derived pigs previously associated with a simple association (SA) or complex association (CA) microbiota were selected at 26 to 37 d of age. In each subject, different jejunal loops were perfused for 8 h with enterotoxigenic K88 (ETEC), ETEC fimbriae (F4), (LAM), or a saline control. The piglets were genotyped for A0 blood group and the glycomic profile was evaluated by microscopic screening of lectin binding: peanut agglutinin (PNA), which is galactose specific; agglutinin I (UEA), which is fucose specific; lectin II (MALii), which is sialic acid specific; concavalin A, which is mannose specific; soybean agglutinin (SBA), which is -acetyl-galactosamine specific; and wheat germ agglutinin (WGA), which is -acetyl-glucosamine specific. A0 pigs had fewer UEA-positive cells, MALii-positive cells ( < 0.001), and SBA-positive cells ( < 0.10) than 00 pigs. Simple association pigs had more SBA positive cells ( < 0.01) than CA pigs. Enterotoxigenic K88-perfused intestinal loops had fewer UEA-positive cells ( < 0.01) and WGA positive cells ( < 0.001) cells and more PNA positive cells (only in SA pigs, < 0.01). No effects of introduction of F4 and LAM in the intestinal lumen were observed. The porcine A0 blood group genotype and the luminal presence of ETEC strongly affected the jejunal mucosa glycomic pattern profile whereas an early oral simple or complex microbial association had limited effects. Pig genetic background has relevance on the cross talk between intestinal epithelium glycocalyx sugar motif and ETEC and, ultimately, on the gut microbial colonization in later life. PMID:27065129

  12. A Novel Lectin with Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Dried Fruiting Bodies of the Monkey Head Mushroom Hericium erinaceum

    Yanrui Li; Guoqing Zhang; Tzi Bun Ng; Hexiang Wang


    A lectin designated as Hericium erinaceum agglutinin (HEA) was isolated from dried fruiting bodies of the mushroom Hericium erinaceum with a chromatographic procedure which entailed DEAE-cellulose, CM-cellulose, Q-Sepharose, and FPLC Superdex 75. Its molecular mass was estimated to be 51 kDa and its N-terminal amino acid sequences was distinctly different from those of other isolated mushroom lectins. The hemagglutinating activity of HEA was inhibited at the minimum concentration of 12...

  13. Acetylcholinesterase-positive afferent axons in mucosa of urinary bladder of adult cats: retrograde tracing and degeneration studies

    Wakabayashi, Y.; Kojima, Y.; Makiura, Y.; Tomoyoshi, T.; Maeda, T.


    Acetylcholinesterase (AchE)-positive afferent axons in the mucosa of the cat urinary bladder were examined in the present experiments. Smallsized dorsal root ganglion cells containing AchE enzyme activity were labelled by injection of retrograde tracer (wheat germ agglutinin conjugated to enzymatically inactive horseradish peroxidase gold complex) into the bladder mucosa of adult cats. Results show that 48.9% (901184) of the labelled ganglion cells possesse...

  14. Anti-M Antibody in Solid Tumors-Two Case Reports

    Soni, Shiv Kumar; Goyal, Hari; Sood, S K; Setia, Rasika


    Anti-M antibodies are usually of IgM, appear as cold agglutinins and are clinically insignificant. We are reporting two cases of anti-M in cases of solid tumors where the anti-M caused discrepancy in blood grouping, reacted in coombs phase of crossmatching. Anti-M in first case showed dosage effect. These antibodies can be clinical significant when detected in coombs phase, making M antigen negative coombs compatible unit transfusion imperative.

  15. [Acute oliguric renal failure and haemolytic anaemia following infectious mononucleosis].

    Brkovic, Natasa; Jørgensen, Kit Riegels; Rosenbæk, Jeppe Bakkestrøm; Pedersen, Erling Bjerregaard


    A 19-year-old man was admitted to hospital due to fatigue, nausea, abdominal pain and faint. He was pale and icteric, awake with sufficient respiration and circulation. He had infectious mononucleosis complicated with acute oliguric renal failure and severe haemolytic anaemia with a positive Coombs test. He had a cold agglutinin syndrome. The treatment comprised intermittent haemodialysis, plasmapheresis and heating. He recovered completely after two months. PMID:26573947

  16. Maintenance of biological activity of pertussis toxin radioiodinated while bound to fetuin-agarose.

    Armstrong, G. D.; Peppler, M S


    We developed a method to produce radioiodinated pertussis toxin (PT) which was active in the goose erythrocyte agglutination and CHO cell assay systems. The procedure used fetuin coupled to agarose to prevent inactivation of the toxin during the iodination reaction. Analysis of the labeled PT by affinity chromatography on fetuin-agarose and wheat germ agglutinin-agarose and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that there were minimal amounts of labeled fetuin...

  17. DBA-Lectin Reactivity Defines Mouse Uterine Natural Killer Cell Subsets with Biased Gene Expression 1

    Chen, Zhilin; Zhang, Jianhong; Hatta, Kota; Lima, Patricia D.A.; Yadi, Hakim; Colucci, Francesco; Yamada, Aureo T.; Croy, B. Anne


    Endometrial decidualization, a process essential for blastocyst implantation in species with hemochorial placentation, is accompanied by an enormous but transient influx of Natural Killer (NK) cells. Mouse uterine (u)NK cell subsets have been defined by diameter and cytoplasmic granule number, reflecting stage of maturity and by histochemical reactivity with Periodic Acid Schiff’s (PAS) reagent, with or without co-reactivity with Dolichos biflorus agglutinin (DBA) lectin. We asked whether DBA...

  18. Optimising expression of the recombinant fusion protein biopesticide ω-hexatoxin-Hv1a /GNA in Pichia pastoris : sequence modifications and a simple method for the generation of multi-copy strains.

    Pyati, P; Fitches, E.; Gatehouse, J A


    Production of recombinant protein bio-insecticides on a commercial scale can only be cost effective if host strains with very high expression levels are available. A recombinant fusion protein containing an arthropod toxin, ω-hexatoxin-Hv1a, (from funnel web spider Hadronyche versuta) linked to snowdrop lectin (Galanthus nivalis agglutinin; GNA) is an effective oral insecticide and candidate biopesticide. However, the fusion protein was vulnerable to proteolysis during production in the yeast...

  19. Accessory Olfactory Bulb Function is Modulated by Input from the Main Olfactory Epithelium

    Slotnick, Burton; Restrepo, Diego; Schellinck, Heather; Archbold, Georgina; Price, Stephen; Lin, Weihong


    While it is now established that sensory neurons in both the main olfactory epithelium and the vomeronasal organ may be activated by both general and pheromonal odorants, it remains unclear what initiates sampling by the VNO. Anterograde transport of wheat germ agglutinin-horseradish peroxidase was used to determine that adequate intranasal syringing with zinc sulfate interrupted all inputs to the main olfactory bulb but left intact those to the accessory olfactory bulb. Adult male treated mi...

  20. Does Candida albicans Als5p Amyloid Play a Role in Commensalism in Caenorhabditis elegans?

    Bois, Michael; Singh, Sean; Samlalsingh, Alyssa; Lipke, Peter N.; Garcia, Melissa C.


    Candida albicans, a dimorphic fungus and an opportunistic pathogen, possesses a myriad of adherence factors, including members of the agglutinin-like sequence (Als) family of mannoproteins. The adhesin Als5p mediates adhesion to many substrates and is upregulated during commensal interactions but is downregulated during active C. albicans infections. An amyloid-forming core sequence at residues 325 to 331 is important for Als5p function, because a single-amino-acid substitution at position 32...

  1. Interleukin 2-dependent and interleukin 2-independent pathways of regulation of thymocyte function by interleukin 6.

    Le, J M; Fredrickson, G.; Reis, L F; Diamantstein, T; Hirano, T.; Kishimoto, T.; Vilcek, J


    Recombinant human interleukin 6 (IL-6), also termed B-cell-stimulatory factor 2 (BSF-2) or interferon-beta 2, was found to stimulate the proliferation of mouse thymocytes costimulated with phytohemagglutinin (PHA). In addition, IL-6 synergistically enhanced the stimulation of thymocyte proliferation by recombinant human interleukin 1 (IL-1) or interleukin 2 (IL-2). Mature thymocytes lacking peanut agglutinin receptor are the main target of IL-6 action. Incubation of thymocytes with IL-6 in th...

  2. Tyrosine-specific protein kinase activity is associated with the purified insulin receptor.

    Kasuga, M.; Fujita-Yamaguchi, Y; Blithe, D L; Kahn, C. R.


    Highly purified human placental insulin receptors were obtained by sequential affinity chromatography on wheat germ agglutinin and insulin-agarose. The preparation had an insulin binding capacity of 4,700 pmol/mg of protein approaching theoretical purity. The purified receptor revealed three major bands of Mr 135,000, 95,000, and 52,000 in NaDodSO4/polyacrylamide gel electrophoresis after reduction by dithiothreitol. All three bands were immunoprecipitated by anti-insulin-receptor antibodies....

  3. Flow cytometry for age assessment of a yeast population and its application in beer fermentations

    Kuřec, Michal; Baszczyňski, Martin; Lehnert, Radek; Mota, André; Teixeira, J.A.; Brányik, Tomáš


    An expeditious method of yeast age estimation was developed based on selective bud scar staining (Alexa Fluor 488-labelled wheat-germ agglutinin) and subsequent fluorescence intensity measurement by flow cytometry. The calibration curve resulting from the cytometric determination of average bud scar fluorescence intensities vs. microscopically counted average bud scar numbers of the same cell populations showed a good correlation and allowed routine cell age estimation by...

  4. Dietary Plant Lectins Appear to Be Transported from the Gut to Gain Access to and Alter Dopaminergic Neurons of Caenorhabditis elegans, a Potential Etiology of Parkinson's Disease.

    Zheng, Jolene; Wang, Mingming; Wei, Wenqian; Keller, Jeffrey N; Adhikari, Binita; King, Jason F; King, Michael L; Peng, Nan; Laine, Roger A


    Lectins from dietary plants have been shown to enhance drug absorption in the gastrointestinal tract of rats, be transported trans-synaptically as shown by tracing of axonal and dendritic paths, and enhance gene delivery. Other carbohydrate-binding protein toxins are known to traverse the gut intact in dogs. Post-feeding rhodamine- or TRITC-tagged dietary lectins, the lectins were tracked from gut to dopaminergic neurons (DAergic-N) in transgenic Caenorhabditis elegans (C. elegans) [egIs1(Pdat-1:GFP)] where the mutant has the green fluorescent protein (GFP) gene fused to a dopamine transport protein gene labeling DAergic-N. The lectins were supplemented along with the food organism Escherichia coli (OP50). Among nine tested rhodamine/TRITC-tagged lectins, four, including Phaseolus vulgaris erythroagglutinin (PHA-E), Bandeiraea simplicifolia (BS-I), Dolichos biflorus agglutinin (DBA), and Arachis hypogaea agglutinin (PNA), appeared to be transported from gut to the GFP-DAergic-N. Griffonia Simplicifolia and PHA-E, reduced the number of GFP-DAergic-N, suggesting a toxic activity. PHA-E, BS-I, Pisum sativum (PSA), and Triticum vulgaris agglutinin (Succinylated) reduced fluorescent intensity of GFP-DAergic-N. PHA-E, PSA, Concanavalin A, and Triticum vulgaris agglutinin decreased the size of GFP-DAergic-N, while BS-I increased neuron size. These observations suggest that dietary plant lectins are transported to and affect DAergic-N in C. elegans, which support Braak and Hawkes' hypothesis, suggesting one alternate potential dietary etiology of Parkinson's disease (PD). A recent Danish study showed that vagotomy resulted in 40% lower incidence of PD over 20 years. Differences in inherited sugar structures of gut and neuronal cell surfaces may make some individuals more susceptible in this conceptual disease etiology model. PMID:27014695

  5. Differential lectin binding on walls of thoraco-cervical blood vessels and lymphatics in rats.

    Kagami, H; Uryu, K; Okamoto, K; Sakai, H; Kaneda, T; Sakanaka, M


    Lectin binding in the walls of large to medium-sized blood vessels and lymphatics in the rat thoraco-cervical region was examined histochemically. The tunica intima of the aorta and superficial cervical artery showed positive reactions with wheat germ agglutinin (WGA) and Concanavalin A (ConA) but not with Dolichus biflorus agglutinin (DBA). The tunica media of the aorta exhibited intense WGA binding, especially on the smooth muscle cells, but the tunica media of the superficial cervical artery did not react with the lectin. Neither ConA nor DBA bound to the tunica media of the aorta and superficial cervical artery. The tunica adventitia of both arteries contained sites binding the three lectins, although DBA reactivity declined as the vascular diameter decreased. The tunica intima of the superior vena cava and azygos vein exhibited positive WGA and ConA binding, whereas DBA binding was noted on only part of the tunica intima of the superior vena cava and not on that of the azygos vein. The tunica media and tunica adventitia were reactive for all three lectins. The WGA and ConA binding sites in the tunica adventitia showed loose networks, suggesting lectin binding on connective tissue elements interlacing among smooth muscle bundles. Lectin binding sites in the walls of lymphatics exhibited an arrangement similar to those in the walls of the veins. Moreover valves protruding into the lumen showed intense WGA and ConA binding and scattered DBA binding. Three other lectins (Ulex europaeus agglutinin, peanut agglutinin, Maclura pomifera) were examined, but they showed no reactions with the vessels. Thus, the differential binding of lectins on the walls of blood vessels and lymphatics of various sizes suggests the functional complexity of monosaccharide residues in the vascular walls. PMID:1758681

  6. Streptococcus mutans Adherence: Presumptive Evidence for Protein-Mediated Attachment Followed by Glucan-Dependent Cellular Accumulation

    Staat, Robert H.; Langley, Sharon D.; Doyle, R J


    Adherence of Streptococcus mutans to smooth surfaces has been attributed to the production of sucrose-derived d-glucans. However, several studies indicate that the bacterium will adhere in the absence of sucrose. The present data confirmed that S. mutans adherence to saliva-coated hydroxyapatite beads in the absence of sucrose is described by the Langmuir equation. The nature of the sucrose-independent adherence was studied with the Persea americana agglutinin as a selective adherence inhibit...

  7. Biochemical characterization, localization and immunostimulating properties of a soluble glycoprotein, Ag1, isolated from in vitro cultures of Plasmodium falciparum

    Jakobsen, P H; Jepsen, S; Riley, E M; Theander, T G; Grellier, P; Lihme, A; Hviid, L; Dziegiel, M; Schrevel, J


    Erythrina christagalli agglutinin, which is specific for carbohydrates bearing beta-D-galactose(1-4)-D-N-acetylglucosamine. Indirect immunofluorescence studies showed that Ag1 is located on the surface of trophozoites and schizonts but not on the surface of merozoites. Ag1 is recognized by human immune sera...... from six different malaria-endemic regions. Ag1 induces in vitro proliferation of lymphocytes from malaria-immune individuals in an antigen-specific manner....

  8. Biochemical characterization, localization and immunostimulating properties of a soluble glycoprotein, Ag1, isolated from in vitro cultures of Plasmodium falciparum

    Jakobsen, P H; Jepsen, S; Riley, E M; Theander, T G; Grellier, P; Lihme, A; Hviid, L; Dziegiel, M; Schrevel, J


    Erythrina christagalli agglutinin, which is specific for carbohydrates bearing beta-D-galactose(1-4)-D-N-acetylglucosamine. Indirect immunofluorescence studies showed that Ag1 is located on the surface of trophozoites and schizonts but not on the surface of merozoites. Ag1 is recognized by human immune sera...... from six different malaria-endemic regions. Ag1 induces in vitro proliferation of lymphocytes from malaria-immune individuals in an antigen-specific manner....

  9. Lectin-binding sites as markers of Golgi subcompartments: proximal-to- distal maturation of oligosaccharides


    We investigated the subcellular sites of glycoprotein oligosaccharide maturation by using lectin conjugates to stain lightly-fixed, saponin- permeabilized myeloma cells. At the electron microscopic level, concanavalin A-peroxidase stains the cisternal space of the nuclear envelope, the rough endoplasmic reticulum, and cisternae along the proximal face of the Golgi stack. Conversely, wheat germ agglutinin- peroxidase stains cisternae along the distal face of the Golgi stack, associated vesicle...

  10. Development and Organization of Ocular Dominance Bands in Primary Visual Cortex of the Sable Ferret

    RUTHAZER, E.S.; Baker, G. E.; Stryker, M.P.


    Thalamocortical afferents in the visual cortex of the adult sable ferret are segregated into eye-specific ocular dominance bands. The development of ocular dominance bands was studied by transneuronal labeling of the visual cortices of ferret kits between the ages of postnatal day 28 (P28) and P81 after intravitreous injections of either tritiated proline or wheat germ agglutinin-horseradish peroxidase. Laminar specificity was evident in the youngest animals studied and was similar to that in...

  11. Direct Production of Ethanol from Raw Corn Starch via Fermentation by Use of a Novel Surface-Engineered Yeast Strain Codisplaying Glucoamylase and α-Amylase

    Shigechi, Hisayori; Koh, Jun; Fujita, Yasuya; Matsumoto, Takeshi; Bito, Yohei; Ueda, Mitsuyoshi; SATOH, Eiichi; Fukuda, Hideki; Kondo, Akihiko


    Direct and efficient production of ethanol by fermentation from raw corn starch was achieved by using the yeast Saccharomyces cerevisiae codisplaying Rhizopus oryzae glucoamylase and Streptococcus bovis α-amylase by using the C-terminal-half region of α-agglutinin and the flocculation functional domain of Flo1p as the respective anchor proteins. In 72-h fermentation, this strain produced 61.8 g of ethanol/liter, with 86.5% of theoretical yield from raw corn starch.

  12. Lectin histochemistry and ultrastructure of feline kidneys from six different storage diseases.

    Castagnaro, M; Alroy, J; Ucci, A A; Glew, R H


    We have compared the pattern of lectin staining with the ultrastructural features of kidneys from normal cats and 19 cats with 6 different lysosomal storage diseases. The diseases studied include GM1 and GM2 gangliosidosis, mucopolysaccharidosis (MPS)-I and MPS-VI, sphingomyelin-lipidosis (i.e., Niemann-Pick disease) and mannosidosis. Ten different biotinylated lectins were used as histochemical probes for carbohydrate residues and avidin-biotin-peroxidase complex as visualant. Concanavalia ensiformis agglutinin (Con A) stained mesangial cells in all storage diseases but GM1, epithelial cells in sphingomyelin-lipidosis and mannosidosis, endothelial cells in GM1 and mannosidosis and Bowman's capsule cells in all but GM2. Griffonia simplicifolia agglutinin I (GS-I) stained the glomerular endothelium in all six diseases, but not in control kidneys. Ricinus communis agglutinin-I (RCA-I) stained the glomerular epithelium only in GM1 and MPS-I. Succinylated wheat germ agglutinin (SWGA) stained the glomerular endothelium and epithelium in mannosidosis, and the glomerular epithelium and Bowman's capsule in MPS-I. Ultrastructure studies demonstrated an accumulation of oligosaccharides in cases of mannosidosis and GM1 gangliosidosis, a mixture of oligosaccharides and lipids in MPS-I, MPS-VI and GM2 gangliosidosis and only lipid storage in sphingomyelin lipidosis. These studies show that morphologic and histochemical changes are manifested in some kidney cell types in lysosomal storage diseases, even though the enzyme deficiency occurs in all cell types. Furthermore, we show that the nature of the undegraded stored material is complex and that other factors, such as rate of membrane turn over, membrane composition, and cell function may influence the amount and nature of the "stored" material. PMID:2892300

  13. Glycophenotype Evaluation in Cutaneous Tumors Using Lectins Labeled with Acridinium Ester

    Luiza Rayanna Amorim Lima


    Full Text Available Background. Tumor cells show alterations in their glycosylation patterns when compared to normal cells. Lectins can be used to evaluate these glycocode changes. Chemiluminescence assay is an effective technique for quantitative analysis of proteins, nucleic acids, and carbohydrates due to its high sensitivity, specificity, and rapid testing. Objective. To use histochemiluminescence based on lectin conjugated to acridinium ester (AE for the investigation of glycophenotype changes in cutaneous tumors. Methods. Concanavalin A (Con A, Peanut agglutinin (PNA, Ulex europaeus agglutinin-I (UEA-I, and Maackia amurensis agglutinin (MAA were conjugated to acridinium ester. Biopsies of cutaneous tumors and normal skin were incubated with the lectins-AE, and chemiluminescence was quantified and expressed as Relative Light Units (RLU. Results. Actinic keratosis (AK, keratoacanthoma (KA, squamous cell carcinoma (SCC, and basal cell carcinoma (BCC showed lower expression of α-D-glucose/mannose and α-L-fucose residues compared to normal tissue. Cutaneous tumors displayed higher expression of Gal-β(1-3-GalNAc residues than normal tissue. AK and SCC exhibited higher expression of Neu5Ac-α(2,3Gal residues than normal epidermis. KA and BCC showed equivalent RLU values compared to normal tissue. Conclusions. Lectin histochemiluminescence allowed quantitative assessment of the carbohydrate expression in cutaneous tissues, contributing to eliminate the subjectivity of conventional techniques used in the histopathological diagnosis.

  14. Fluorescence emission and polarization analyses for evaluating binding of ruthenium metalloglycocluster to lectin and tetanus toxin c-fragment

    Okada, Tomoko; Minoura, Norihiko


    We have developed a fluorescent ruthenium metalloglycocluster as a powerful molecular probe for evaluating a binding event between carbohydrates and lectins by fluorescence emission (FE) and fluorescence polarization (FP) analysis. The fluorescent ruthenium metalloglycoclusters, [Ru(bpy-2Gal)3] and [Ru(bpy-2Glc)3], possess clustered galactose and glucose surrounding the ruthenium center. Changes in FE and FP of these metalloglycoclusters were measured by adding each lectin (Peanut agglutinin (PNA), Ricinus communis agglutinin 120 (RCA), Concanavalin A (ConA), or Wheat germ agglutinin (WGA)) or tetanus toxin c-fragment (TCF). Following the addition of PNA, the FE spectrum of [Ru(bpy- 2Gal)3] showed new emission peak and the FP value of [Ru(bpy-2Gal)3] increased. Similarly, the FE spectrum of [Ru(bpy-2Glc)3] showed new emission peak and the FP value increased following the addition of ConA. Since other combinations of the metalloglycoclusters and lectin caused little change, specific bindings of galactose to PNA and glucose to ConA were proved by the FE and FP measurement. From nonlinear least-squares fitting, dissociation constants (Kd) of [Ru(bpy-2Gal)3] to PNA was 6.1 μM, while the Kd values of [Ru(bpy)2(bpy-2Gal)] to PNA was ca. 10-4 M. Therefore, the clustered carbohydrates were proved to increase affinity to lectins. Furthermore, the FP measurements proved specific binding of [Ru(bpy-2Gal)3] to TCF.

  15. Lectin interactions with alpha-galactosylated xenoantigens

    Kirkeby, Svend; Moe, Dennis


    alpha-Galactosylated xenoantigens (Galalpha1-3Galbeta1-4GlcNAcbeta1 and Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc) are often detected with the alpha-Gal specific lectin Griffonia simplicifolia 1 isolectin B4 (GS1 B4). However, this lectin exhibits a broad and variable specificity for......-galactophilic lectins to alpha-galactosylated neoglycoproteins. The lectins were: Euonymus europaeus agglutinin (EEA), Griffonia simplicifolia 1 isolectin B4 (GS1 B4), Maclura pomifera agglutinin (MPA) and Pseudomonas aeruginosa agglutinin (PA-IL). Although both GS1 B4 and MPA strongly bound glycoconjugates terminating...... in Gal there seems to be some differentiation in their sugar binding preferences. MPA was the only lectin that showed high affinity for the pentasaccharide Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc and for the Galalpha-glycans on non-primate thyroglobulin. The length of the xenoantigenic...

  16. Fluorescence emission and polarization analyses for evaluating binding of ruthenium metalloglycoclusters to lectins and tetanus toxin C-fragment

    Okada, Tomoko; Minoura, Norihiko


    We develop a fluorescent ruthenium metalloglycocluster for use as a powerful molecular probe in evaluating the binding between carbohydrates and lectins by fluorescence emission (FE) and fluorescence polarization (FP) analyses. Changes in the FE and FP of these metalloglycoclusters are measured following the addition of lectin [peanut agglutinin (PNA), Ricinus communis agglutinin 120, Concanavalin A (ConA), or wheat germ agglutinin] or tetanus toxin c-fragment (TCF). After the addition of PNA, the FE spectrum of [Ru(bpy-2Gal)3] shows a new emission peak and the FP value of [Ru(bpy-2Gal)3] increases. Similarly, the FE spectrum of [Ru(bpy-2Glc)3] shows a new emission peak and the FP value increases on addition of ConA. Because other combinations of metalloglycoclusters and lectins show little change, specific binding of galactose to PNA and that of glucose to ConA are confirmed by the FE and FP measurements. Resulting dissociation constants (Kd) prove that the metalloglycoclusters with highly clustered carbohydrates show higher affinity for the respective lectins than those with less clustered carbohydrates. Furthermore, specific binding of [Ru(bpy-2Gal)3] to TCF was confirmed by the FP measurement.

  17. Effect of Bu-Zhong-Yi-Qi-Tang on deficiency of N-glycan/nitric oxide and islet damage induced by streptozotocin in diabetic rats

    Xiao-Qiu Liu; Ling Wu; Xue-Jun Guo


    AIM: To investigate the effect of Bu-Zhong-Yi-Qi-Tang (Decoction for Reinforcing Middle Jiao and Replenishing Qi) on deficiency of N-glycan/nitric oxide (NO) and islet damage induced by injecting two medium doses of streptozotocin (STZ). METHODS: Diabetes was induced by intraperitoneal injection of STZ at 55 mg/kg on day 1 and day 8. Islet damage was evaluated using a scoring system. Nitrite, nitrate, α-mannosidase and amylase activities were measured by colorimetry. N-glycan patterns of amylase were determined with lectin [ConA, pisum sativum agglutinin (PSA), peanut agglutinin (PNA), and lens culinaris agglutinin (LCA)] affinity precipitation method. RESULTS: Severe islet necrosis and mild islet atrophy were observed in diabetic rats. The number and size of islets, the activities of α-mannosidase, amylase and nitrite were decreased, while the binding of PNA and LCA to amylase was increased. All of which were improved after treatment with Bu-Zhong-Yi-Qi-Tang. Islet damage was significantly correlated with nitrite, nitrate, α-mannosidase, amylase and the binding of LCA, PNA, and PSA to amylase. PNA, and PSA to amylase.CONCLUSION: STZ- induced i s let damage i s related to N-glycan def iciency in proteins by blocking α-mannosidase activity and no deficiency, accumulation of unfolded proteins, and endoplasmic reticulum stress and activation of cellular signals, all of which are improved after treatment with Bu-Zhong-Yi- Qi-Tang.

  18. Targeting the Cryptococcus neoformans var. grubii Cell Wall Using Lectins: Study of the Carbohydrate-Binding Domain

    Pamella de Brito Ximenes


    Full Text Available Cryptococcus neoformans var. grubii is considered to be the major cause of cryptococcosis in immunosuppressed patients. Understanding cell wall glycoproteins using lectins is of medical interest and can contribute to specific therapy. The aim of this study was to evaluate the carbohydrates on the cell wall of Cryptococcus neoformans var. grubii clinical isolates, using a fluorescein isothiocyanate-lectin binding protocol. Thirty yeast strains stocked in the culture collection were cultivated for 2 days at 30 °C with shaking. Cells were obtained by centrifugation, washed in phosphate-buffered saline, and a suspension of 107 cells/mL was obtained. To determine the binding profile of lectins, concanavalin A (Con A, wheat germ agglutinin (WGA, Ulex europaeus agglutinin I (UEA-I, and peanut agglutinin (PNA conjugated to fluorescein were used. All the tested clinical isolates of Cryptococcus neoformans var. grubii were intensely stained by WGA, moderately stained by Con A, and weakly stained by PNA and UEA-I. Thus, Cryptococcus can be detected in clinical specimens such as blood and cerebrospinal fluid using the fluorescent lectin WGA, which may be considered as an option for detection in cases of suspected cryptococcosis with low laboratory sensitivity. Future applications may be developed using this basic tool.

  19. Prevalence of Salmonella typhi among Patients in Abeokuta, South-Western Nigeria

    I.O. Okonko


    Full Text Available This study reports on the prevalence of typhoid fever between genders among patients in Abeokuta, Nigeria. Typhoid fever caused by Salmonella typhi is an endemic disease in the tropic and sub-tropic and has become a major public health problem in developing countries of the world with an estimated annual incidence of 540 per 100,000. The annual incidence of typhoid is estimated to be about 17 million cases worldwide. It is often encountered in tropical countries including Nigeria where they constitute serious sources of morbidities and mortalities. Blood samples were collected from 840 apparently healthy people; 460 (54.8% females and 380 (45.2% males. The samples were examined for the presence and levels of Salmonella typhi antibodies by Widal agglutination technique. The standard Samonella ‘O’ and ‘H’ suspension (ANTEC diagnostic products were used as antigens. Of the 840 sera tested, agglutinins to Salmonella typhi were most prevalent in female subjects accounting for [426(92.6%] of the ‘H’ antigens and [322(70.0%] of ‘O’ antigens at the various dilutions while in the male subjects, [351(92.4%] accounts for the ‘O��� and [327(86.1%] for the ‘H’ antigens. There was a female preponderance (F/M 2:1. The levels of agglutinin of Salmonella paratyphi C-H [93(24.5% ] and Salmonella typhi C-O [112(29.5%] in the males were however, low. In the females, the low significant agglutinin titres for Salmonella typhi O and Salmonella paratyphi A-O were observed in 27.6% and 36.1% of the sera respectively. The results of this study showed that more males had Salmonella agglutinin titres for S. typhi O [351(92.4%] and S. typhi H [327(86.1%]. More so, 132 (34.7% males had Salmonella agglutinin titres for S. paratyphi A-O, 119 (31.3% for S. paratyphi B-O, 112 (29.5% for S. paratyphi C-O, 117 (31.0% for S. paratyphi A-H, 125 (33.0% for S. paratyphi B-H, and 93 (24.5% for S. paratyphi C-H. It also showed that more females had Salmonella

  20. Real-time cytotoxicity assay for rapid and sensitive detection of ricin from complex matrices.

    Diana Pauly

    Full Text Available BACKGROUND: In the context of a potential bioterrorist attack sensitive and fast detection of functionally active toxins such as ricin from complex matrices is necessary to be able to start timely countermeasures. One of the functional detection methods currently available for ricin is the endpoint cytotoxicity assay, which suffers from a number of technical deficits. METHODOLOGY/FINDINGS: This work describes a novel online cytotoxicity assay for the detection of active ricin and Ricinus communis agglutinin, that is based on a real-time cell electronic sensing system and impedance measurement. Characteristic growth parameters of Vero cells were monitored online and used as standardized viability control. Upon incubation with toxin the cell status and the cytotoxic effect were visualized using a characteristic cell index-time profile. For ricin, tested in concentrations of 0.06 ng/mL or above, a concentration-dependent decrease of cell index correlating with cytotoxicity was recorded between 3.5 h and 60 h. For ricin, sensitive detection was determined after 24 h, with an IC50 of 0.4 ng/mL (for agglutinin, an IC50 of 30 ng/mL was observed. Using functionally blocking antibodies, the specificity for ricin and agglutinin was shown. For detection from complex matrices, ricin was spiked into several food matrices, and an IC50 ranging from 5.6 to 200 ng/mL was observed. Additionally, the assay proved to be useful in detecting active ricin in environmental sample materials, as shown for organic fertilizer containing R. communis material. CONCLUSIONS/SIGNIFICANCE: The cell-electrode impedance measurement provides a sensitive online detection method for biologically active cytotoxins such as ricin. As the cell status is monitored online, the assay can be standardized more efficiently than previous approaches based on endpoint measurement. More importantly, the real-time cytotoxicity assay provides a fast and easy tool to detect active ricin in complex

  1. Adenovirus-mediated expression of pig α(1, 3) galactosyltransferase reconstructs Gal α(1, 3) Gal epitope on the surface of human tumor cells


    Gal α(1,3)Gal(gal epitope)is a carbohydrate epitope and synthesized in large amount by α(1,3)galactosyltransferase [α(1,3)GT] enzyme on the cells of lower mammalian animals such as pigs and mice.Human has no gal epitope due to the inactivation of α(1,3)GT gene but produces a large amount of antibodies(anti-Gal)which recognize Gal α(1,3)Gal structures specifically.In this study,a replicationdeficient recombinant adenoviral vector Ad5sGT containing pig α(1,3)GT cDNA was constructed and characterized.Adenoviral vector-mediated transfer of pig α(1,3)GT gene into human tumor cells such as malignant melanoma A375,stomach cancer SGC-7901,and lung cancer SPC-A-1 was reported for the first time.Results showed that Gal epitope did not increase the sensitivity of human tumor cells to human complement-mediated lysis,although human complement activation and the binding of human IgG and IgM natural antibodies to human tumor cells were enhanced significantly after Ad5sGT transduction.Appearance of gal epitope on the human tumor cells changed the expression of cell surface carbohydrates reacting with Ulex europaeus I(UEA I)lectins,Vicia villosa agglutinin(VVA),Arachis hypogaea agglutinin(PNA),and Glycine max agglutinin(SBA)to different degrees.In addition,no effect of gal epitope on the growth in vitro of human tumor cells was observed in MTT assay.

  2. Dietary plant lectins appear to be transported from the gut to gain access to and alter dopaminergic neurons of Caenorhabditis elegans, a potential etiology of Parkinson’s disease

    Jolene eZheng


    Full Text Available Lectins from dietary plants have been shown to enhance drug absorption in the gastrointestinal tract of rats, be transported trans-synaptically as shown by tracing of axonal and dendritic paths, and enhance gene delivery. Other carbohydrate-binding protein toxins are known to traverse the gut intact in dogs. Post-feeding rhodamine- or TRITC-tagged dietary lectins, the lectins were tracked from gut to dopaminergic neurons (DAergic-N in transgenic Caenorhabditis elegans (C. elegans (egIs1[Pdat-1::GFP] where the mutant has the Green Fluorescent Protein (GFP gene fused to a dopamine transport protein gene labeling dopaminergic neurons, The lectins were supplemented along with the food organism Escherichia coli (OP50. Among nine tested rhodamine/TRITC-tagged lectins, four, including Phaseolus vulgaris erythroagglutinin (PHA-E, Bandeiraea simplicifolia (BS-I, Dolichos biflorus agglutinin (DBA, and Arachis hypogaea (PNA, appeared to be transported from gut to the GFP-DAergic-N. Griffonia Simplicifolia (GSL-I and PHA-E, reduced the number of GFP-DAergic-N suggesting a toxic activity. PHA-E, BS-I, Pisum Sativum (PSA, and Triticum vulgaris agglutinin (Succinylated reduced fluorescent intensity of GFP-DAergic-N. PHA-E, PSA, Concanavalin A, and Triticum vulgaris agglutinin decreased the size of GFP-DAergic-N, while BS-I increased neuron size. These observations suggest that dietary plant lectins are transported to and affect DAergic-N in C. elegans, which support Braak and Hawkes’ hypothesis, suggesting one alternate potential dietary etiology of Parkinson’s disease (PD. A recent Danish study showed that vagotomy resulted in 40% lower incidence of PD over 20 years. Differences in inherited sugar structures of gut and neuronal cell surfaces may make some individuals more susceptible in this conceptual disease etiology model.

  3. Lectin binding and effects in culture on human cancer and non-cancer cell lines: examination of issues of interest in drug design strategies.

    Petrossian, Karineh; Banner, Lisa R; Oppenheimer, Steven B


    By using a non-cancer and a cancer cell line originally from the same tissue (colon), coupled with testing lectins for cell binding and for their effects on these cell lines in culture, this study describes a simple multi-parameter approach that has revealed some interesting results that could be useful in drug development strategies. Two human cell lines, CCL-220/Colo320DM (human colon cancer cells, tumorigenic in nude mice) and CRL-1459/CCD-18Co (non-malignant human colon cells) were tested for their ability to bind to agarose microbeads derivatized with two lectins, peanut agglutinin (Arachis hypogaea agglutinin, PNA) and Dolichos biflorus agglutinin (DBA), and the effects of these lectins were assessed in culture using the MTT assay. Both cell lines bound to DBA-derivatized microbeads, and binding was inhibited by N-acetyl-D-galactosamine, but not by L-fucose. Neither cell line bound to PNA-derivatized microbeads. Despite the lack of lectin binding using the rapid microbead method, PNA was mitogenic in culture at some time points and its mitogenic effect displayed a reverse-dose response. This was also seen with effects of DBA on cells in culture. While this is a simple study, the results were statistically highly significant and suggest that: (1) agents may not need to bind strongly to cells to exert biological effects, (2) cell line pairs derived from diseased and non-diseased tissue can provide useful comparative data on potential drug effects and (3) very low concentrations of potential drugs might be initially tested experimentally because reverse-dose responses should be considered. PMID:17706752

  4. Isolation and partial characterization of a D-galactose-binding lectin from the latex of Synadenium carinatum

    Maria Aparecida de Souza; Francielle Amâncio-Pereira; Cristina Ribeiro de Barros Cardoso; Adriano Gomes da Silva; Edmar Gomes Silva; Lívia Resende Andrade; Janethe Deolina Oliveira Pena; Henrique Lanza; Sandra Regina Afonso-Cardoso


    A lectin from the latex of Synadenium carinatum was purified by affinity chromatography on immobilized-D-galactose-agarose and shown to be a potent agglutinin of human erythrocytes. The haemagglutination of human red cells was inhibited by 3.0 mM N-acetyl-D-galactopyranoside, 6.3 mM methyl-beta-D-galactopyranoside, 50 mM methyl-alpha-D-galactopyranoside and 50 mM D-fucose but not by L-fucose, demonstrating an anomeric and a conformational specificity. According to SDS-PAGE analysis, the lecti...

  5. Effect of lectins on mouse peritoneal macrophage phagocytic activity.

    Maldonado, G; Porras, F; Fernández, L; Vázquez, L; Zenteno, E


    We studied the in vitro ability of lectin-treated murine peritoneal macrophages to attach and phagocytize particulate antigens. Glucose and mannose specific lectins such as Con-A and lentil lectin, as well as complex lactosamine residues specific lectins, such as Phaseolus vulgaris var. cacahuate and Phaseolus coccineus var. alubia, increased the macrophage phagocytic activity towards heterologous erythrocytes, whereas peanut agglutinin, a galactose-specific lectin, diminished the macrophage phagocytic activity. These results suggest that a galactose-N-acetyl-D galactosamine-containing structure could participate as negative modulator of the phagocytic activity. PMID:7851961

  6. Display of wasp venom allergens on the cell surface of Saccharomyces cerevisiae

    Borodina, Irina; Jensen, B. M.; Søndergaard, Ib;


    Saccharomyces cerevisiae preserve their native allergenic properties and whether the yeast native surface glycoproteins interfere with IgE binding. We chose to use the major allergens from the common wasp Vespula vulgaris venom: phospholipase A1, hyaluronidase and antigen 5 as the model. Results: The proteins...... were expressed on the surface as fusions with a-agglutinin complex protein AGA2. The expression was confirmed by fluorescent cytometry (FACS) after staining the cells with antibody against a C-tag attached to the C-terminal end of the allergens. Phospholipase A1 and hyaluronidase retained their...

  7. Functional activities of the Tsh protein from avian pathogenic Escherichia coli (APEC) strains

    Renata K. T. Kobayashi; Gaziri, Luis Carlos J.; Vidotto, Marilda C.


    The temperature-sensitive hemagglutinin (Tsh) expressed by strains of avian pathogenic Escherichia (E.) coli (APEC) has both agglutinin and protease activities. Tsh is synthesized as a 140 kDa precursor protein, whose processing results in a 106 kDa passenger domain (Tshs) and a 33 kDa β-domain (Tshβ). In this study, both recombinant Tsh (rTsh) and supernatants from APEC, which contain Tshs (106 kDa), caused proteolysis of chicken tracheal mucin. Both rTsh (140 kDa) and pellets from wild-type...


    Qian Yang; Ling-shu Wan; Zhi-kang Xu


    A glycopolymer bearing glucose residues was tethered onto the surface of polypropylene microporous membrane by UV-induced graft polymerization of α-allyl glucoside. Concanavalin A (Con A), a glucose recognizing lectin, could be specifically adsorbed to the membrane surface. On the other hand, the membrane surface showed no recognition ability to another lectin peanut agglutinin. Moreover, the recognition complex between the glycosylated membrane surface and Con Acould be inhibited by glucose and mannose solution. This surface glycosylated membrane could be used as affinity membrane for protein separation and purification.


    WANG Xiao-mei; SHAN Jun; CHEN Zhuo-huai


    Aim: To investigate glycoconjugate changes on the cell surface of proliferative lesions and neoplasms of mice lungs at various stages of tumorigenesis, the relation between progressive development of mouse pulmonary tumors and expression of cell surface saccharide. Materials and methods: Thirty - one male A/J strain mice at 5 weeks of age were treated intraperitoneally with a single injection of 20 - methylcholanthrene (20 - MC), 292 pulmonary lesions including 31 hyperplasias, 145 alveolar adenomas, 61 papillary adenomas, 55 papillary adenocarcinomas and their combined type were obtained. The binding affinities of cells in normal respiratory epithelia and in proliferative lesions to four peroxidases - conjugated lectins, Maclura pomifera agglutinin (MPA), Arachis hypogea agglutinin (PNA), Ricinus communis agglutinin (RCA), and wheat germ agglutinin (WGA) were examined. Results: Cells of hyperplasia and alveolar adenoma showed fairly strong affinity to all the four lectins. However, part of papillary adenoma cells and greater part of papillary adenocarcinoma cells lost their binding affinity to MPA, PNA, and RCA, but not to WGA. The bindings of MPA, PNA and RNA were detected predominently on the luminal surfaces of benign tumors but not on the luminal surfaces of malignant tumors. WGA might bind to varied types of benign and malignant tumors. Pretreated with neuraminidase, the lesions enhanced the staining intensity for the four lectins, the binding sites of WGA to malignant tumor cells were numerous. A distinct difference in lectin binding affinity between hyperplasia / alveolar adenoma/papillary adenoma and papillary adenocarcinoma was clearly shown( x2 = 46.89, P < 0.01, x2 = 36.77, P < 0.01 and x2 = 52.87, P < 0.01 ) in this experiment. The complex glycoconjugates on the cell surface of malignant and benign lesions during the development of pulmonary tumor were changed,malignant tumor cells differed from the surface of benign tumor cells, the levels of

  10. Brain alpha-dystroglycan displays unique glycoepitopes and preferential binding to laminin-10/11.

    McDearmon, Erin L; Combs, Ariana C; Sekiguchi, Kiyotoshi; Fujiwara, Hironobu; Ervasti, James M


    alpha-Dystroglycan was quantitatively enriched from mammalian brain based on its uniform reactivity with Vicia villosa agglutinin and resolved into sub-populations possessing or lacking the sulfated glucuronic acid epitope recognized by monoclonal antibody HNK-1. We generated a new monoclonal antibody specific for a glycoepitope on brain alpha-dystroglycan but absent from alpha-dystroglycan expressed in all other tissues examined. Finally, we found that laminin-10/11 preferentially bound to brain alpha-dystroglycan compared to skeletal muscle alpha-dystroglycan. Our results suggest that tissue-specific glycosylation modifies the laminin binding specificity of alpha-dystroglycan. PMID:16709410

  11. Synthesis of amides and sulfonamides of β-D- galactopyranosylamine and β-lactosylamine and evaluation of their interactions with the lectins from Erythrina cristagalli and Ricinus communis

    We report herein the synthesis of some β-D-galactopyranosylamine and β-lactosylamine amides and sulfonamides. The interactions of these compounds with lectins from the seeds of Erythrina cristagalli (LEC) and Ricinus communis (RCA120) were evaluated in a hemagglutination inhibitory activity assay. D-Galactose and lactose were used as reference compounds. The β-lactosylamine amides and sulfonamides were nearly as active as lactose in inhibiting LEC mediated hemagglutination and were less active against RCA120 agglutinin. The β-D-galactopyranosylamine amides and sulfonamides were, with one exception, considerably less active than D-galactose in the assay with both lectins. (author)

  12. [Seroepidemiologic survey of leptospirosis among environmental sanitation workers in an urban locality in the southern region of Brazil].

    de Almeida, L P; Martins, L F; Brod, C S; Germano, P M


    Sera from 386 environmental sanitation workers, concerned with water supply, drains and drainage galleries, sewers, garbage collection and road sweepers, were examined for leptospiral agglutinins by the microscopic agglutination test. Altogether 40 of the 386 workers (10.4%) were positive to one or more serovars; however, the difference in seropositivity between the professional categories was not significant (p 0.05). Of the seropositive workers, 86.9% had agglutination titres > or = 100 and < or = 400; the rates for titres 100 and 400 were higher than 800, 1,600 and 3,200 (p < 0.05). PMID:7997826

  13. Cytostructural Localization of a Tumor-Associated Antigen

    Howard, Donald R.; Batsakis, John G.


    Tumor cell membrane glycoproteins may be involved in the induction of tumor immunity or in the escape of tumors from immunologic defense mechanisms. Forty-four benign and malignant breast lesions were examined for the presence of a carbohydrate precursor antigen (T antigen) of the human blood group system MN. T antigen was demonstrated by means of an immunohistochemical technique to detect tissue binding of peanut agglutinin, a plant lectin, with affinity for T antigen. Malignant breast lesions showed a pattern of T antigen expression different from that of benign breast tissues. A possible role for T antigen in the modulation of the immune response to breast carcinoma is suggested.

  14. Enhancement of bone marrow allografts from nude mice into mismatched recipients by T cells void of graft-versus-host activity.

    Lapidot, T; Lubin, I; Terenzi, A; Faktorowich, Y; Erlich, P; Reisner, Y


    Transplantation of 8 x 10(6) C57BL/6-Nu+/Nu+ (nude) bone marrow cells into C3H/HeJ recipients after conditioning with 8 Gy of total body irradiation has resulted in a markedly higher rate of graft rejection or graft failure compared to that found in recipients of normal C57BL/6 or C57BL/6-Bg+/Bg+ (beige) T-cell-depleted bone marrow. Mixing experiments using different numbers of nude bone marrow cells with or without mature thymocytes (unagglutinated by peanut agglutinin) revealed that engraft...

  15. Altered glycosylation, expression of serum haptoglobin and alpha-1-antitrypsin in chronic hepatitis C, hepatitis C induced liver cirrhosis and hepatocellular carcinoma patients.

    Mondal, Gautam; Saroha, Ashish; Bose, Partha Pratim; Chatterjee, B P


    Liver cirrhosis with hepatitis C viral infection (HCV-LC) causes high risk to develop hepatocellular carcinoma (HCC). Besides diagnosis of liver cirrhosis by biochemical test, imaging techniques, assessment of structural liver damage by biopsy shows several disadvantages. Our aim was to monitor the changes in the expression level of serum proteins and their glycosylation pattern among chronic hepatitis C (HCV-CH), HCV-LC and HCC patients with respect to controls. 2D gel electrophoresis of HCV-CH, HCV-LC and HCC patients' sera showed several protein spots, which were identified by LC-MS. The change in the expression of two prominent protein spots, haptoglobin (Hp) and alpha 1-antitrypsin (AAT) was evaluated by western blot and ELISA. The changes in glycosylation pattern of these serum proteins were assayed using different lectins. Increased level of Hp and AAT was observed in HCV-LC and HCC patients' group whereas those were found to be present less in HCV-CH patient groups with respect to control as determined by ELISA using monoclonal antibodies. Decreased level of sialylation in both Hp and AAT was observed in HCV-LC and HCV-CH patients' group whereas increased level of sialylation was observed in HCC patient groups by ELISA using Sambucus nigra agglutinin. On the other hand increased level of fucosylation in two serum glycoproteins was observed in HCV-LC and HCC patients' group using Lens culinarris agglutinin. High glycan branching was found in HCV-LC and HCC patient groups in Hp but not in HCV-CH as determined by Datura stramonium agglutinin. However, there was no such change observed in glycan branching in AAT of HCV-CH and HCV-LC patients' groups, to the contrary high glycan branching was observed in HCC patients' group. Increased level of exposed galactose in both serum proteins was observed in both HCC patients' group as determined by Ricinus communis agglutinin. The present glycoproteomics study could predict the progression of HCV-CH, HCV-LC and HCC

  16. Mouse whey acidic protein is a novel member of the family of 'four-disulfide core' proteins.

    Hennighausen, L G; Sippel, A E


    Unlike in other mammalian species, the major whey protein in mouse is not alpha-lactalbumin, but a cysteine rich, acidic protein with a molecular weight of 14.0 kDa. We have deduced the amino acid sequence of this mouse acidic of whey protein from the nucleotide sequence of cloned cDNA. The positions of the half cysteines suggest that mouse whey acidic protein (WAP) is a two domain protein, very similar in structure to the plant lectin wheat germ agglutinin and the hypothalamic carrier protei...

  17. Gum and Nose Bleeding as a Presentation of Pediatric Brucellosis



    Full Text Available Introduction Brucellosis symptoms are nonspecific; the most common complaints include fever, sweats, anorexia, headache, malaise, and arthralgia. Hematological manifestations of active brucellosis vary from mild anemia and leukopenia to thrombocytopenia and rarely pancytopenia. Case Presentation We report on an eight-year-old boy who presented epistaxis and gum bleeding. The physical examination revealed petechiae, purpura, ecchymosis, and cervical while inguinal lymphadenopathy and splenomegaly were noted. Brucella agglutinin titer was positive. After five days of specific therapy for brucellosis, fever was controlled, clinical signs and symptoms were improved and platelet count was dramatically increased. Conclusions Sever thrombocytopenia and bleeding may be the presentation of brucellosis.

  18. Investigation of the response to the enterobacterial common antigen after typhoid vaccination

    Arlete M. Milhomem


    Full Text Available Antibodies against the Salmonella typhi enterobacterial common antigen (ECA and the O and H antigens were investigated in sera from healthy male subjects who had been previously vaccinated with the typhoid vaccine. No serological response to ECA was observed. Sera from subjects not previously vaccinated presented titers of ECA hemagglutinins which quantitatively were related to the presence ofH titers, but not to O agglutinins but with no statistical significance. The results are discussed in relation to the possible protective immunological mechanisms in typhoid fever.

  19. Langerhans Cell Histiocytosis of the Temporal Bone.

    Ginat, Daniel Thomas; Johnson, Daniel N; Cipriani, Nicole A


    Langerhans cell histiocytosis involving the temporal bone region is uncommon and can resemble malignant neoplasms on imaging due to high cellularity. Although recognizing the presence of sharp margins with beveled-edges can be helpful, tissue sampling is often necessary for confirming the diagnosis. Cytology classically demonstrates kidney-bean shaped nuclei within the Langerhans cells and immunohistochemical staining is positive for S-100, peanut agglutinin (PNA), MHC class II, CD1a, and Langerin (CD 207). These features are exemplified in this sine qua non radiology-pathology correlation article. PMID:25903273

  20. Equestrian perniosis: a report of 2 cases and a review of the literature.

    Stewart, Campbell L; Adler, Donald J; Jacobson, Abby; Brod, Bruce A; Shinohara, Michi M; Seykora, John T; Elenitsas, Rosalie; Rosenbach, Misha


    Equestrian perniosis (EP) is a rare condition in which patients develop tender burning nodular plaques on their bilateral thighs after riding in the cold. These lesions tend to resolve rapidly with minimal exposure to cold, and wearing loose, layered warm clothing. Unlike acral perniosis, EP has no known systemic disease associations, although 2 reported cases did have elevated cold agglutinins. The histology of this disease is similar to perniosis; however, EP is distinct in that the perivascular lymphocytic infiltrate prominently involves the fat. In this case report, we discuss the clinical and histological findings in 2 cases of EP, including the first documented in a man. PMID:22534636


    Oey Kam Nio


    Full Text Available Penelitian sebelumnya menunjukkan adanya suatu zat toksis pada biji saga yang tidak tergolong faktor "anti-nutritive" seperti "tripsin inhibitors", "phyto-agglutinins"dan "saponins". Ternyata bahwa zat toksis tersebut yang belum dapat diekstraksi dan diidentifikasi, berada di dalam fraksi minyak biji saga. Di samping adanya zat toksis, ditemukan pub bahwa kualitas protein biji saga lebih rendah dari­pada kacang kedele. Hasil analisa asam amino menunjukkan bahwa kadar methionine dan threonine ada­lah terbatas (limiting, apabila ditambah dengan kedua asam amino ini dalam jumlah yang cukup, kuali­tas proteinnya menjadi sama tingginya dengan kacang kedele rebus tanpa tambahan.

  2. Design of a covalently bonded glycosphingolipid microarray

    Arigi, Emma; Blixt, Klas Ola; Buschard, Karsten;


    agglutinin, a monoclonal antibody to sulfatide, Sulph 1; and a polyclonal antiserum reactive to asialo-G(M2)). Preliminary evaluation of the method indicated successful immobilization of the GSLs, and selective binding of test probes. The potential utility of this methodology for designing covalent...... 2-mercaptoethylamine, was also tested. Underivatized or linker-derivatized lyso-GSL were then immobilized on N-hydroxysuccinimide- or epoxide-activated glass microarray slides and probed with carbohydrate binding proteins of known or partially known specificities (i.e., cholera toxin B-chain; peanut...

  3. Rapid reconstitution of a transmembrane protein into supported planar lipid membranes.

    Nakanishi, M


    A procedure for reconstituting a transmembrane protein by the freeze-thaw method into supported planar lipid layers has been developed. A solution containing human glycophorin A was introduced between an alkylated cover glass with lipid layers from soybean phospholipids and a bare glass slide, and was then put in a glass dish which was frozen outside by liquid nitrogen. The lipid layer membranes prepared in this manner have been examined by the binding of both macrophages and wheat germ agglutinin agarose. Macrophages bound more efficiently to the membranes bearing glycophorin A and spread more rapidly than those of the control membranes. PMID:6548452

  4. Direct production of ethanol from raw corn starch via fermentation by use of a novel surface-engineered yeast strain codisplaying glucoamylase and alpha-amylase.

    Shigechi, Hisayori; Koh, Jun; Fujita, Yasuya; Matsumoto, Takeshi; Bito, Yohei; Ueda, Mitsuyoshi; Satoh, Eiichi; Fukuda, Hideki; Kondo, Akihiko


    Direct and efficient production of ethanol by fermentation from raw corn starch was achieved by using the yeast Saccharomyces cerevisiae codisplaying Rhizopus oryzae glucoamylase and Streptococcus bovis alpha-amylase by using the C-terminal-half region of alpha-agglutinin and the flocculation functional domain of Flo1p as the respective anchor proteins. In 72-h fermentation, this strain produced 61.8 g of ethanol/liter, with 86.5% of theoretical yield from raw corn starch. PMID:15294847

  5. Lectin typing of Campylobacter jejuni using a novel quartz crystal microbalance technique

    Yakovleva, Maria E., E-mail: [Department of Infectious Diseases and Medical Microbiology, Lund University, 223 62 Lund (Sweden); Moran, Anthony P. [Department of Microbiology, School of Natural Sciences, National University of Ireland, Galway (Ireland); Safina, Gulnara R. [Department of Analytical and Marine Chemistry, University of Gothenburg, 412 96 Gothenburg (Sweden); Wadstroem, Torkel [Department of Infectious Diseases and Medical Microbiology, Lund University, 223 62 Lund (Sweden); Danielsson, Bengt [Acromed Invest AB, Magistratsvaegen 10, 226 43 Lund (Sweden)


    Seven Campylobacter jejuni strains were characterised by a lectin typing assay. The typing system was based on a quartz crystal microbalance technique (QCM) with four commercially available lectins (wheat germ agglutinin, Maackia amurensis lectin, Lens culinaris agglutinin, and Concanavalin A), which were chosen for their differing carbohydrate specificities. Initially, the gold surfaces of the quartz crystals were modified with 11-mercaptoundecanoic acid followed by lectin immobilisation using a conventional amine-coupling technique. Bacterial cells were applied for lectin typing without preliminary treatment, and resonant frequency and dissipation responses were recorded. The adhesion of microorganisms on lectin surfaces was confirmed by atomic force microscopy. Scanning was performed in the tapping mode and the presence of bacteria on lectin-coated surfaces was successfully demonstrated. A significant difference in the dissipation response was observed for different C. jejuni strains which made it possible to use this parameter for discriminating between bacterial strains. In summary, the QCM technique proved a powerful tool for the recognition and discrimination of C. jejuni strains. The approach may also prove applicable to strain discrimination of other bacterial species, particularly pathogens.

  6. Assessment of lectin inactivation by heat and digestion.

    Pusztai, A; Grant, G


    Proteins/glycoproteins from plants, particularly lectins, are more resistant to heat denaturation than animal proteins (1, 2). With legume seeds, whose lectin content is appreciable, this presents potentially serious problems in nutritional practice. Therefore, before they can be used safely, legume-based food/ feeds usually require thorough and expensive heat processing to inactivate antinutritive components. Indeed, dry or moist heating of seeds at 70°C for several h has little or no effect on their lectin activity (Fig. 1) and treatment at much higher temperatures is needed to inactivate the biological and antinutritional effects of legume lectins (1, 2). The safety aspect is even more serious with some monocot lectins, such as wheatgerm agglutinin or a number of oilseed lectins, such as peanut agglutinin and many others because they are extremely heat stable and normal cooking or other conventional heat treatments may fail to inactivate them (3) Thus, the best way to avoid potential harmful effects of these heat-resistant lectins is to limit their dietary intake to a minimum. Fig. 1. Loss of lectin activity during aqueous heat treatment of soybean at various temperatures. PMID:21374488

  7. Density variant glycan microarray for evaluating cross-linking of mucin-like glycoconjugates by lectins.

    Godula, Kamil; Bertozzi, Carolyn R


    Interactions of mucin glycoproteins with cognate receptors are dictated by the structures and spatial organization of glycans that decorate the mucin polypeptide backbone. The glycan-binding proteins, or lectins, that interact with mucins are often oligomeric receptors with multiple ligand binding domains. In this work, we employed a microarray platform comprising synthetic glycopolymers that emulate natural mucins arrayed at different surface densities to evaluate how glycan valency and spatial separation affect the preferential binding mode of a particular lectin. We evaluated a panel of four lectins (Soybean agglutinin (SBA), Wisteria floribunda lectin (WFL), Vicia villosa-B-4 agglutinin (VVA), and Helix pomatia agglutin (HPA)) with specificity for α-N-acetylgalactosamine (α-GalNAc), an epitope displayed on mucins overexpressed in many adenocarcinomas. While these lectins possess the ability to agglutinate A(1)-blood cells carrying the α-GalNAc epitope and cross-link low valency glycoconjugates, only SBA showed a tendency to form intermolecular cross-links among the arrayed polyvalent mucin mimetics. These results suggest that glycopolymer microarrays can reveal discrete higher-order binding preferences beyond the recognition of individual glycan epitopes. Our findings indicate that glycan valency can set thresholds for cross-linking by lectins. More broadly, well-defined synthetic glycopolymers enable the integration of glycoconjugate structural and spatial diversity in a single microarray screening platform. PMID:22967056

  8. Binucleate trophoblast giant cells in the water buffalo (Bubalus bubalis) placenta.

    Carvalho, A F; Klisch, K; Miglino, M A; Pereira, F T V; Bevilacqua, E


    The binucleate trophoblast giant cells (BNC) of the water buffalo, Bubalus bubalis, placenta were studied, with emphasis on the synthesis of BNC-specific proteins. Placentomal tissues of 27 water buffalos (2-10 months of pregnancy) were processed for light and electron microscopy. The frequency of BNCs was 20% of the trophoblastic cells in 2-3-month placentas and increased to 27% in the later stages. Ultrastructurally, binucleate cells displayed a prominent granular endoplasmic reticulum and Golgi apparatus, typical of cells involved with protein synthesis and exportation. The buffalo BNCs contained periodic acid-Schiff (PAS)-positive granules and reacted with antisera against bovine placental lactogen, prolactin-related protein-I, and pregnancy-associated glycoproteins. Lectin histochemistry with Dolichos biflorus agglutinin, Vicia villosa agglutinin, and Phaseolus vulgaris leucoagglutinin showed specific staining of BNCs. Different stages of BNC migration and fusion with uterine epithelial cells were observed. Trinucleate feto-maternal hybrid cells were the typical outcome of cell fusions. These cells underwent degeneration, with typical morphological features of apoptosis. The results revealed a strong homology between water buffalo and cattle BNCs concerning cell morphology, protein expression, glycosylation pattern, and characteristics of cell migration and fusion. PMID:16240388

  9. The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study.

    Jones, Carolyn J P; Wilsher, Sandra A; Wooding, F B P; Benirschke, K; Allen, W R


    The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. Some differences between Okapi and Giraffe were evident. Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. PMID:25527317

  10. Microarrays with varying carbohydrate density reveal distinct subpopulations of serum antibodies.

    Oyelaran, Oyindasola; Li, Qian; Farnsworth, David; Gildersleeve, Jeffrey C


    Antigen arrays have become important tools for profiling complex mixtures of proteins such as serum antibodies. These arrays can be used to better understand immune responses, discover new biomarkers, and guide the development of vaccines. Nevertheless, they are not perfect and improved array designs would enhance the information derived from this technology. In this study, we describe and evaluate a strategy for varying antigen density on an array and then use the array to study binding of lectins, monoclonal antibodies, and serum antibodies. To vary density, neoglycoproteins containing differing amounts of carbohydrate were synthesized and used to make a carbohydrate microarray with variations in both structure and density. We demonstrate that this method provides variations in density on the array surface within a range that is relevant for biological recognition events. The array was used to evaluate density dependent binding properties of three lectins (Vicia villosa lectin B4, Helix pomatia agglutinin, and soybean agglutinin) and three monoclonal antibodies (HBTn-1, B1.1, and Bric111) that bind the tumor-associated Tn antigen. In addition, serum antibodies were profiled from 30 healthy donors. The results show that variations in antigen density are required to detect the full spectrum of antibodies that bind a particular antigen and can be used to reveal differences in antibody populations between individuals that are not detectable using a single antigen density. PMID:19366269

  11. Distribution patterns of influenza virus receptors and viral attachment patterns in the respiratory and intestinal tracts of seven avian species

    Costa Taiana


    Full Text Available Abstract This study assessed the presence of sialic acid α-2,3 and α-2,6 linked glycan receptors in seven avian species. The respiratory and intestinal tracts of the chicken, common quail, red-legged partridge, turkey, golden pheasant, ostrich, and mallard were tested by means of lectin histochemistry, using the lectins Maackia amurensis agglutinin II and Sambucus nigra agglutinin, which show affinity for α-2,3 and α-2,6 receptors, respectively. Additionally, the pattern of virus attachment (PVA was evaluated with virus histochemistry, using an avian-origin H4N5 virus and a human-origin seasonal H1N1 virus. There was a great variation of receptor distribution among the tissues and avian species studied. Both α-2,3 and α-2,6 receptors were present in the respiratory and intestinal tracts of the chicken, common quail, red-legged partridge, turkey, and golden pheasant. In ostriches, the expression of the receptor was basically restricted to α-2,3 in both the respiratory and intestinal tracts and in mallards the α-2,6 receptors were absent from the intestinal tract. The results obtained with the lectin histochemistry were, in general, in agreement with the PVA. The differential expression and distribution of α-2,3 and α-2,6 receptors among various avian species might reflect a potentially decisive factor in the emergence of new viral strains.

  12. Glycan-mediated uptake in urothelial primary cells: Perspectives for improved intravesical drug delivery in urinary tract infections.

    Pichl, Clara Maria; Feilhauer, Sophie; Schwaigerlehner, Rose-Marie; Gabor, Franz; Wirth, Michael; Neutsch, Lukas


    Urinary tract infections (UTIs) are among the most common bacterial infections. Despite a wide range of therapeutic options, treatment success is compromised by multiresistance and the efficient mechanism of tissue colonization of uropathogenic Escherichia coli (UPEC). In advanced drug delivery systems, a similar, glycan-mediated targeting mechanism may be realized by conjugating the drug to a plant lectin. This may lead to the drug being more efficiently accumulated at the desired site of action, the bacterial reservoirs. In this study, we aimed at elucidating the potential of this biorecognitive approach. Glycan-triggered interaction cascades and uptake processes of several plant lectins with distinct carbohydrate specificities were characterized using single cells and monolayer culture. Due to pronounced cytoadhesive and cytoinvasive properties, wheat germ agglutinin (WGA) emerged as a promising targeter in porcine urothelial primary cells. The lectin-cell interaction proved highly stabile in artificial urine, simulating the conditions in actual application. Colocalisation studies with internalized WGA and lens culinaris agglutinin (LCA) revealed that intracellular accumulation sites were largely identical for GlcNAc- and Mannose-specific lectins. This indicates that WGA-mediated delivery may indeed constitute a potent tool to reach bacteria taken up via a FimH-triggered invasion process. Existing pitfalls in intravesical treatment schedules may soon be overcome. PMID:26383837

  13. Local cholinergic and non-cholinergic neural pathways to the rat supraoptic nucleus

    An estimated two thirds of the input to the supraoptic nucleus of the rat hypothalamus (SON) including a functionally significant cholinergic innervation, arise from local sources of unknown origin. The sources of these inputs were identified utilizing Golgi-Cox, retrograde tracing, choline acetyltransferase immunocytochemistry and anterograde tracing methodologies. Multipolar Golgi impregnated neurons located dorsal and lateral to the SON extend spiney processes into the nucleus. Injections of the retrograde tracers, wheat germ agglutinin or wheat germ agglutinin-horseradish peroxidase, into the SON labeled cells bilaterally in the arcuate nucleus, and ipsilaterally in the lateral hypothalamus, anterior hypothalamus, nucleus of the diagonal band, subfornical organ, medial preoptic area, lateral preoptic area and in the region dorsolateral to the nucleus. Immunocytochemistry for choline acetyltransferase revealed cells within the ventro-caudal portion of cholinergic cell group, Ch4, which cluster dorsolateral to the SON, and extend axon- and dendrite-like processes into the SON. Cells double-labeled by choline acetyltransferase immunocytochemistry and retrograde tracer injections into the SON are localized within the same cholinergic cell group dorsolateral to the SON. Injections of the anterograde tracer, Phaseolus vulgaris-leucoagglutinin, deposited dorsolateral to the SON results in labeled pre-and post-synaptic processes within the SON. The identification and characterization of endogenous immunoglobulin within the SON and other neurons innervating areas lacking a blood-brain barrier established a novel and potentially important system for direct communication of the supraoptic cells with blood-borne constitutents

  14. Differences in cellular glycoconjugates of quiescent, inflamed, and neoplastic colonic epithelium in colitis and cancer-prone tamarins.

    Moore, R; King, N; Alroy, J


    In the preceding paper the authors demonstrated that the lectin staining patterns of normal colonic epithelium obtained from colitis and carcinoma-prone cotton top tamarins (CTTs), Saguinus oedipus, a New World primate, differs from colitis- and carcinoma-resistant primate species. In this study they determined the usefulness of cytochemical features in inflamed epithelium as indicators for malignant change. They compared the lectin staining pattern in inflamed mucosa and adjacent mucosa with colonic carcinoma from 8 CTTs with that of 9 clinically healthy CTTs with no histologic evidence of colitis. Deparaffinized sections were labeled with ten biotinylated lectins and stained by the avidin-biotin peroxidase complex method. Numerous significant differences were demonstrated in the lectin staining pattern between normal epithelium and colonic carcinoma; fewer between normal and chronic inflamed epithelium. However, between chronic inflamed epithelium and colonic carcinoma significant staining differences were observed with only two lectins, peanut agglutinin (PNA) and Ulex europaeus agglutinin-I (UEA-I). These findings suggest that there is a progression in alteration of lectin staining pattern from normal epithelium, via chronic colitis, to colonic carcinoma. Furthermore, the differences between chronic colitis and colonic carcinoma are expressed only with those lectins that are associated with malignant transformation of human colonic epithelium. PMID:3132858

  15. Identification and preliminary characterization of a Ca2+-dependent hemagglutinin in the celomic fluid of Sipunculus nudus

    L Ballarin


    Full Text Available A soluble agglutinin was purified by affinity chromatography of the celomic fluid of the marine worm Sipunculus nudus. This agglutinin requires metal cations for its activity and is specific for derivatives of D-galactose. It resulted lightly thermostable, with a pH optimum around 7.5. On SDS-PAGE, it was resolved in two bands, of 33 and 31 kDa in reducing conditions and 29 and 26 kDa in non-reducing conditions. This behavior is probably due to the presence of disulfide bridges between cysteine residues, which are required for the correct functioning of the hemagglutinin, as β-mercaptoethanol completely abolish the agglutinating activity of cell-free celomic fluid. The purified lectin can influence in vitro phagocytosis of yeast by celomic leukocytes: in the presence of the molecule, ingestion of foreign particles results significantly decreased and yeast cells agglutinate and forms rosettes around the celomocytes. This suggests a role of the molecule in immunosurveillance.

  16. Anatomy, histochemistry and immunohistochemistry of the olfactory subsystems in mice

    Arthur William Barrios


    Full Text Available The four regions of the murine nasal cavity featuring olfactory neurons were studied anatomically and by labelling with lectins and relevant antibodies with a view to establishing criteria for the identification of olfactory subsystems that are readily applicable to other mammals. In the main olfactory epithelium and the septal organ the olfactory sensory neurons (OSNs are embedded in quasi-stratified columnar epithelium; vomeronasal OSNs are embedded in epithelium lining the medial interior wall of the vomeronasal duct and do not make contact with the mucosa of the main nasal cavity; and in Grüneberg’s ganglion a small isolated population of OSNs lies adjacent to, but not within, the epithelium. With the exception of Grüneberg’s ganglion, all the tissues expressing olfactory marker protein (OMP (the above four nasal territories, the vomeronasal and main olfactory nerves, and the main and accessory olfactory bulbs are also labelled by Lycopersicum esculentum agglutinin, while Ulex europaeus agglutinin I labels all and only tissues expressing Gi2 (the apical sensory neurons of the vomeronasal organ, their axons, and their glomerular destinations in the anterior accessory olfactory bulb. These staining patterns of UEA-I and LEA may facilitate the characterization of olfactory anatomy in other species. A 710-section atlas of the anatomy of the murine nasal cavity has been made available on line.

  17. Anatomy, histochemistry, and immunohistochemistry of the olfactory subsystems in mice.

    Barrios, Arthur W; Núñez, Gonzalo; Sánchez Quinteiro, Pablo; Salazar, Ignacio


    The four regions of the murine nasal cavity featuring olfactory neurons were studied anatomically and by labeling with lectins and relevant antibodies with a view to establishing criteria for the identification of olfactory subsystems that are readily applicable to other mammals. In the main olfactory epithelium and the septal organ the olfactory sensory neurons (OSNs) are embedded in quasi-stratified columnar epithelium; vomeronasal OSNs are embedded in epithelium lining the medial interior wall of the vomeronasal duct and do not make contact with the mucosa of the main nasal cavity; and in Grüneberg's ganglion a small isolated population of OSNs lies adjacent to, but not within, the epithelium. With the exception of Grüneberg's ganglion, all the tissues expressing olfactory marker protein (OMP) (the above four nasal territories, the vomeronasal and main olfactory nerves, and the main and accessory olfactory bulbs) are also labeled by Lycopersicum esculentum agglutinin, while Ulex europaeus agglutinin I labels all and only tissues expressing Gαi2 (the apical sensory neurons of the vomeronasal organ, their axons, and their glomerular destinations in the anterior accessory olfactory bulb). These staining patterns of UEA-I and LEA may facilitate the characterization of olfactory anatomy in other species. A 710-section atlas of the anatomy of the murine nasal cavity has been made available on line. PMID:25071468

  18. Evaluation of the antischistosomal activity of sulfated α-D-glucan from the lichen Ramalina celastri free and encapsulated into liposomes

    R.V.S. Araújo


    Full Text Available The antischistosomal activity of the sulfated polysaccharide α-D-glucan (Glu.SO4 extracted from Ramalina celastri was evaluated after encapsulation into liposomes (Glu.SO4-LIPO in Schistosoma mansoni-infected mice. The effect of treatment with Glu.SO4 and Glu.SO4-LIPO (10 mg/kg on egg elimination, worm burden and hepatic granuloma formation was assessed using female albino Swiss mice, 35-40 days of age, weighing 25 ± 2 g, infected with 150 cercariae/animal (Biomphalaria glabrata, BH strain. Four groups (N = 10 were studied, two controls (empty liposomes and NaCl and two treated groups (Glu.SO4-LIPO and Glu.SO4 using a single dose. Parasitological analysis revealed that Glu.SO4-LIPO was as efficient as Glu.SO4 in reducing egg elimination and worm burden. Treatment with free Glu.SO4 and Glu.SO4-LIPO induced a statistically significant reduction in the number of granulomas (62 and 63%, respectively. Lectin histochemistry showed that wheat germ agglutinin intensely stained the egg-granuloma system in all treated groups. On the other hand, peanut agglutinin stained cells in the control groups, but not in the treated groups. The present results suggest a correlation between the decreasing number of hepatic egg-granulomas and the glycosylation profile of the egg-granuloma system in animals treated with free Glu.SO4 or Glu.SO4-LIPO.

  19. Affinity Separation of Lectins Using Porous Membranes Immobilized with Glycopolymer Brushes Containing Mannose or N-Acetyl-D-Glucosamine

    Yoshiko Miura


    Full Text Available Porous membranes with glycopolymer brushes were prepared as biomaterials for affinity separation. Glycopolymer brushes contained acrylic acid and D-mannose or N-acetyl-D-glucosamine, and were formed on substrates by surface-initiated atom transfer radical polymerization. The presence of glycopolymer brush was confirmed by X-ray photoelectron spectroscopy, contact angle, and ellipsometry measurements. The interaction between lectin and the glycopolymer immobilized on glass slides was confirmed using fluorescent-labeled proteins. Glycopolymer-immobilized surfaces exhibited specific adsorption of the corresponding lectin, compared with bovine serum albumin. Lectins were continuously rejected by the glycopolymer-immobilized membranes. When the protein solution was permeated through the glycopolymer-immobilized membrane, bovine serum albumin was not adsorbed on the membrane surface. In contrast, concanavalin A and wheat germ agglutinin were rejected by membranes incorporating D-mannose or N-acetyl-D-glucosamine, respectively. The amounts of adsorbed concanavalin A and wheat germ agglutinin was increased five- and two-fold that of adsorbed bovine serum albumin, respectively.

  20. The effect of the state of differentiation on labeling of epidermal cell surface glycoproteins

    Epidermal cells were grown in a medium in which the Ca++ concentration controlled the stage of differentiation. Cell surface molecules of differentiated and undifferentiated cells were compared by lactoperoxidase-catalyzed iodination, by the interaction with 125I-lectins, and by the metabolic incorporation of L-(3H)-fucose. Molecular weights of the labeled components were determined by SDS-PAGE and autoradiography. After lactoperoxidase iodination, most of the radioactivity was found in polypeptide bands of 79,000, 65,000 and 56,000 daltons. The 79,000 band is the most intense for undifferentiated cells but disappears as differentiation proceeds. The 56,000 band is present in normal cells at all stages of differentiation but is absent from neoplastic cells. Glycoproteins reacted with 125I-lectins were found at 180,000, 130,000 and 85,000 daltons. The 130,000 band was the most prominent for differentiated cells labeled with wheat germ agglutinin but was essentially absent from the undifferentiated cells. With Ricinus communis agglutinin, this band was weaker for undifferentiated than for differentiated cells but was the most intense for both. After metabolic incorporation of tritiated fucose, radioactive glycoproteins were found at 130,000 and 85,000 daltons, with comparable intensities for differentiated and undifferentiated cells

  1. Direct targeting of cancer cells: a multiparameter approach.

    Heinrich, Eileen L; Welty, Lily Anne Y; Banner, Lisa R; Oppenheimer, Steven B


    Lectins have been widely used in cell surface studies and in the development of potential anticancer drugs. Many past studies that have examined lectin toxicity have only evaluated the effects on cancer cells, not their non-cancer counterparts. In addition, few past studies have evaluated the relationship between lectin-cell binding and lectin toxicity on both cell types. Here we examine these parameters in one study: lectin-cell binding and lectin toxicity with both cancer cells and their normal counterparts. We found that the human colon cancer cell line CCL-220/Colo320DM bound to agarose beads derivatized with Phaseolus vulgaris agglutinin (PHA-L) and wheat germ agglutinin (WGA), while the non-cancer human colon cell line CRL-1459/CCD-18Co did not. When these lectins were tested for their effects on cell viability in culture, both cell lines were affected by the lectins but at 6, 48 and 72 h incubation times, PHA-L was most toxic to the cancer cell line in a concentration dependent manner. At 48 h incubation, WGA was more toxic to the cancer cell line. The results suggest that it may be possible to develop lectin protocols that selectively target cancer cells for death. In any case, examination of both malignant cells and their non-malignant counterparts, analysis of their binding characteristics to immobilized lectins, and examination of the toxicity of free lectins in culture, provides a multiparameter model for obtaining more comprehensive information than from more limited approaches. PMID:16181664

  2. Partial isolation and characterisation of a hemagglutinating factor from avocado seed.

    Yaakobovich, Y; Neeman, I


    Extracts of ground avocado seeds (Fuerte and Hass varieties), prepared in different buffer solutions (pH 2.0-12.0), show hemagglutinating activity towards A, B, AB, and H (0) human erythrocytes. The extract showing the highest titer of aggulination was extracted at pH 10.5. The crude extract also causes hemolysis of fresh washed erythrocytes. The hemagglutinating factor is not inhibited by most of the simple sugars tested, e.g., D-glucose, D-mannose, D-galactose, and glucose-amine. The only sugars which show some inhibitory effect are N-Acetyl-neuraminic acid, melibiose, and stachiose. Basic amino acids, e.g., lysine and arginine also inhibit its activity. However the most potent inhibitors of the agglutinin are proteins and glycoproteins such as bovine serum albumin, collagen, thyroglobulin, ovalbumin, mucin, and fetuin. The agglutinin is adsorbed on polymer beads such as Sepharose 4B, Sephadex G100, Agarose, and Chitin, and it reacts with hog erythrocyte membranes. It can be partially eluted from those materials with alkaline buffers (pH 9.0-10.5). PMID:6578749

  3. A kinetic analysis of the tumor-associated galactopyranosyl-(1→3)-2-acetamido-2-deoxy--D-galactopyranoside antigen-lectin interaction

    Bandaru Narasimha Murthy; Narayanaswamy Jayaraman


    A kinetic study of the tumor-associated galactopyranosyl-(1→3)-2-acetamido-2-deoxy--Dgalactopyranoside (T-antigen) with lectin peanut agglutinin is described. The disaccharide antigen was synthesized by chemical methods and was functionalized suitably for immobilization onto a carboxymethylated sensor chip. The ligand immobilized surface was allowed interaction with the lectin peanut agglutinin, which acted as the analyte and the interaction was studied by the surface plasmon resonance method. The ligand-lectin interaction was characterized by the kinetic on-off rates and a bivalent analyte binding model was found to describe the observed kinetic constants. It was identified that the antigen-lectin interaction had a faster association rate constant (a1) and a slower dissociation rate constant (d1) in the initial binding step. The subsequent binding step showed much reduced kinetic rates. The antigen-lectin interaction was compared with the kinetic rates of the interaction of a galactopyranosyl-(1→ 4)--D-galactopyranoside derivative and a mannopyranoside derivative with the lectin.

  4. Design of near-infrared fluorescent bioactive conjugated functional iron oxide nanoparticles for optical detection of colon cancer

    Corem-Salkmon E


    Full Text Available Enav Corem-Salkmon, Benny Perlstein, Shlomo MargelThe Institute of Nanotechnology and Advanced Materials, Department of Chemistry, Bar-Ilan University, Ramat-Gan, IsraelBackground: Colon cancer is one of the major causes of death in the Western world. Early detection significantly improves long-term survival for patients with the disease. Near-infrared (NIR fluorescent nanoparticles hold great promise as contrast agents for tumor detection. NIR offers several advantages for bioimaging compared with fluorescence in the visible spectrum, ie, lower autofluorescence of biological tissues, lower absorbance, and consequently deeper penetration into biomatrices.Methods and results: NIR fluorescent iron oxide nanoparticles with a narrow size distribution were prepared by nucleation, followed by controlled growth of thin iron oxide films onto cyanine NIR dye conjugated gelatin-iron oxide nuclei. For functionalization, and in order to increase the NIR fluorescence intensity, the NIR fluorescent iron oxide nanoparticles obtained were coated with human serum albumin containing cyanine NIR dye. Leakage of the NIR dye from these nanoparticles into phosphate-buffered saline solution containing 4% albumin was not detected. The work presented here is a feasibility study to test the suitability of iron oxide-human serum albumin NIR fluorescent nanoparticles for optical detection of colon cancer. It demonstrates that encapsulation of NIR fluorescent dye within these nanoparticles significantly reduces photobleaching of the dye. Tumor-targeting ligands, peanut agglutinin and anticarcinoembryonic antigen antibodies (αCEA, were covalently conjugated with the NIR fluorescent iron oxide-human serum albumin nanoparticles via a poly(ethylene glycol spacer. Specific colon tumor detection was demonstrated in chicken embryo and mouse models for both nonconjugated and the peanut agglutinin-conjugated or αCEA-conjugated NIR fluorescent iron oxide-human serum albumin

  5. Childhood brucellosis--a microbiological, epidemiological and clinical study.

    Mantur, B G; Akki, A S; Mangalgi, Smita S; Patil, S V; Gobbur, R H; Peerapur, B V


    A total of 5726 blood specimens (from children aged 14 years and younger) were studied for the serological evidence of brucellosis. Ninety-three (1.6 per cent) showed diagnostic agglutinin titres with a geometric mean titre of 403 (SD +/- 547). Forty-three (59.7 per cent) blood specimens yielded the growth of Brucella melitensis. Thirty-nine patients (41.93 per cent) were shepherds, who constituted the major occupational group affected in the present series. More than 60 per cent of the patients had a history of both consumption of fresh goat's milk and close animal contact. The habit of consuming fresh goat's milk to obtain relief from chronic ailments was noted in nine patients. Seventy-three (78.49 per cent) were males and 20 (21.51 per cent) were females, with a male to female ratio of 3:1. The disease occurred mainly in the school age group (mean age 10.3 years). All the patients had an acute history of less than 2 months. Forty-nine (52.68 per cent) patients presented with persistent fever, 19 (20.43 per cent) with joint pain, and the rest with a combination of fever and joint pain with and without low backache, fever being the commonest complaint. One case presented with involuntary movements of limbs alone and the other with burning feet only. Pityriasis alba was the consistent physical finding, with fever in the majority of the patients. The major joint found to be involved was the knee (52.77 per cent). The synovial fluid obtained from the knee joint of five patients demonstrated Brucella agglutinins and also three grew B. melitensis. Eight patients presented with complications that included skin lesions (3), carditis (2), neurobrucellosis such as chorea (1), peripheral neuritis (1), and meningitis (1). Brucella melitensis biotype 1 was successfully isolated from the papular eruption of one out of three cases who presented with skin lesions. To our knowledge this is the fourth confirmed isolation of B. melitensis from skin lesions with brucellosis

  6. Syntheses of Sulfo-Glycodendrimers Using Click Chemistry and Their Biological Evaluation

    Tomohiro Fukuda


    Full Text Available A series of novel glycol-clusters containing sulfonated N-acetyl-D-glucosamine (GlcNAc have been synthesized using click chemistry. Three dendrimers with aromatic dendrons were synthesized using chlorination, azidation and click chemistries. The resulting dendrimers were modified with azide-terminated sulfonated GlcNAc using click chemistry. The sulfonated dendrimers showed affinity for proteins, including the lectin wheat germ agglutinin and amyloid beta peptide (1-42. The dendrimers of G1 and G2 in particular showed the largest affinity for the proteins. The addition of the sulfonated GlcNAc dendrimers of G1 and G2 exhibited an inhibition effect on the aggregation of the amyloid beta peptide, reduced the b-sheet conformation, and led to a reduction in the level of nanofiber formation.

  7. PNA: a marker of neoplastic progression and differentiation in the gastro-intestinal tract.

    Grigolato, P; Benetti, A; Berenzi, A; Villanacci, V; Tardanico, R


    We examined 35 cases of stomach carcinoma and 40 cases of colonic carcinoma with PNA associated with peroxidase (peanut agglutinin, lectin which binds to the terminal disaccharide galactose beta (1,3)-N-acetil-galacto-samine). In this way evaluation of the functional aspects of the normal-neoplastic sequence was undertaken. This method was carried out for histological and ultrastructural investigations. The results obtained in both cases showed a different reactivity in the evolution of neoplastic disease: in fact, positivity in dysplasia is finely granular intracytoplasmic, whereas in well-differentiated neoplastic transformation such a reactivity is preferentially localized along the cellular membranes, with restoration of gross positivity in the cytoplasm for the poorly-differentiated neoplasm. We therefore believe PNA to be a marker not only of neoplastic progression but of differentiation as well: we also hypothesize it to reveal glycoprotein groups with possible antigenic power, involved in immunologic interactions between tumor and host. PMID:2283482

  8. Títulos de anticorpos aglutinantes induzidos por vacinas comerciais contra leptospirose bovina Agglutinating antibody titers induced by commercial vaccines against bovine leptospirosis

    Gabriela de Godoy Cravo Arduino


    serovars. All vaccines used were capable to product agglutinins for the Hardjo and Wolffi serovars observed at 3 days after vaccination, remaining until the 150th day; those serovars induced the highest titres of agglutinins. Vaccine D, in spite of not containing the Wolffi serovar, induced the production of agglutinins to this serovar. Agglutinins to the Canicola serovar were only observed in the animals vaccinated with the D bacterine. Vaccine D induced the highest average titers of antibodies to all tested serovars.

  9. Visual detection of serum asialohaptoglobin by plasmonic sandwich ELLSA--a new platform for cirrhosis diagnosis.

    Bose, Partha Pratim; Mandal, Gautam; Kumar, Dharmendra; Duseja, Ajay; Chatterjee, Bishnu Pada


    The cirrhotic condition of the liver has long been acknowledged as the preface to liver cancer. The desialylation status of the serum acute phase protein, haptoglobin, has been introduced as a new diagnostic analyte for liver cirrhosis. The reliability of this new diagnostic molecule has been evaluated in 30 liver cirrhosis patients having a history of earlier viral hepatitis C (HCV-LC). A novel enzyme linked lectinosorbent assay has been developed coupled with the plasmon mechanism of gold nanoparticle aggregation as the colorimetric read out which can visually distinguish the cirrhotic liver patients from the normal healthy and hepatitis C controls. The assay can be useful for rapid point-of-care detection, and even an untrained person can execute it without a specialized instrument. This method employs Sambucus nigra agglutinin (SNA) to detect the extent of α-2,6 sialylation of serum haptoglobin, the new diagnostic molecule for liver cirrhosis. PMID:26568048

  10. Detection of Sugar-Lectin Interactions by Multivalent Dendritic Sugar Functionalized Single-Walled Carbon Nanotubes

    Vasu, K S; Bagul, R S; Jayaraman, N; Sood, A K; 10.1063/1.4739793


    We show that single walled carbon nanotubes (SWNT) decorated with sugar functionalized poly (propyl ether imine) (PETIM) dendrimer is a very sensitive platform to quantitatively detect carbohydrate recognizing proteins, namely, lectins. The changes in electrical conductivity of SWNT in field effect transistor device due to carbohydrate - protein interactions form the basis of present study. The mannose sugar attached PETIM dendrimers undergo charge - transfer interactions with the SWNT. The changes in the conductance of the dendritic sugar functionalized SWNT after addition of lectins in varying concentrations were found to follow the Langmuir type isotherm, giving the concanavalin A (Con A) - mannose affinity constant to be 8.5 x 106 M-1. The increase in the device conductance observed after adding 10 nM of Con A is same as after adding 20 \\muM of a non - specific lectin peanut agglutinin, showing the high specificity of the Con A - mannose interactions. The specificity of sugar-lectin interactions was chara...