Lectins

Lectins are group of proteins or glycoprotein of non immunological origins, which can recognize specific carbohydrate structure. In this research work hemaggulutination was used as technique for screening Bangladeshi vegetables for lectin. Five varieties of legume plant seeds Canavalia gladiata(Sword bean),Lens culinaris (Moshordal), Peasum sativam (Motorsuti), Vigna unguiculata subsp. sesquipedalis (Borboti), Cajanus cajan (Arhordal), one Amaranthaceae Amaranthus caudate (katoadata) and one fruit Citrullus lanatus (Watermelon) species from Bangladesh were examined for lectins with chicken and human erythrocytes. Crude extracts from all the species showed agglutinating activity against the erythrocytes used. The lowest protein concentration required to produce erythrocytes agglutination varied remarkable ranging from 0.7 µg/ml to 8080 µg/ml. The strongest activities were shown in the agglutination of human blood erythrocytes by partial purification of lectin from Moshordal and chick...

Human erythrocyte sialoglycoproteins, or glycophorins, were chromatographed by lectin-affinity and metal-interaction chromatography on high-performance liquid chromatographic columns. Glycophorins A, B and C were separated from other proteins and from glycophorin E by using a column containing wheat germ agglutinin, immobilized on a microparticulate silica support. The glycophorins were adsorbed on the lectin column from a mobile phase containing 0.25 A4 sodium chloride and recovered by stepwise desorption with 0.2 M N-acetylglucosamine solution. Glycophorins A, B and C were separated into the individual components on a silica-bound iminodiacetic acid stationary phase in the copper(H) chelate form. The separation of the glycophorins by metal-interaction chromatography was accomplished by decreasing salt gradient elution. Retention times and resolution of the individual glycophorins were sensitive to the initial sodium chloride concentration and the pH of the eluent. Addition of methanol to the eluent increased the resolution. The effects of linear, decreasing gradients of pH and methanol in 25 rnM phosphate buffer on the resolution of glycophorins were also investigated. In both types of chromatography the mobile phases contained 0.05% (w/v) sodium dodecyl sulfate. With octylglycoside or CHAPS in the eluent glycophorins A and C could not be eluted. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to analyze all the chromatographic results.

a glycosylated arginase acting as a fungal lectin from Peltigera canina is able to produce recruitment of cyanobiont Nostoc cells and their adhesion to the hyphal surface. This implies that the cyanobiont would develop organelles to motility toward the chemoattractant. however when visualized by transmission electron microscopy, Nostoc cells recently isolated from P. canina thallus do not reveal any motile, superficial organelles, although their surface was covered by small spindles and serrated layer related to gliding. The use of S-(3,4-dichlorobenzyl)isothiourea, blebbistatin, phalloidin and latrunculin a provide circumstantial evidence that actin microfilaments rather than MreB, the actin-like protein from prokaryota, and probably, an aTPase which develops contractile function similar to that of myosin II, are involved in cell motility. These experimental facts, the absence of superficial elements (fimbriae, pili or flagellum) related to cell movement, and the appearance of sunken cells during of after movement verified by scanning electron microscopy, support the hypothesis that the motility of lichen cyanobionts could be achieved by contraction-relaxation episodes of the cytoskeleton induced by fungal lectin act as a chemoattractant.

Most commercial yeast strains are nonflocculent. However, controlled flocculation phenotypes could provide significant benefits to many fermentation-based industries. In nonflocculent laboratory strains, it has been demonstrated that it is possible to adjust flocculation and adhesion phenotypes to desired specifications by altering expression of the otherwise silent but dominant flocculation (FLO) genes. However, FLO genes are characterized by high allele heterogeneity and are subjected to epigenetic regulation. Extrapolation of data obtained in laboratory strains to industrial strains may therefore not always be applicable. Here, we assess the adhesion phenotypes that are associated with the expression of a chromosomal copy of the FLO1, FLO5, or FLO11 open reading frame in two nonflocculent commercial wine yeast strains, BM45 and VIN13. The chromosomal promoters of these genes were replaced with stationary phase-inducible promoters of the HSP30 and ADH2 genes. Under standard laboratory and wine making conditions, the strategy resulted in expected and stable expression patterns of these genes in both strains. However, the specific impact of the expression of individual FLO genes showed significant differences between the two wine strains and with corresponding phenotypes in laboratory strains. The data suggest that optimization of the flocculation pattern of individual commercial strains will have to be based on a strain-by-strain approach.

The infectious and diagnostic stage of Giardia lamblia (also known as G. intestinalis or G. duodenalis) is the cyst. The Giardia cyst wall contains fibrils of a unique b-1,3-linked N-acetylgalactosamine (GalNAc) homopolymer and at least three cyst wall proteins (CWPs) composed of Leu-rich repeats (CWP LRR) and a C-terminal conserved Cys-rich region (CWP CRR). Our goals were to dissect the structure of the cyst wall and determine how it is disrupted during excystation. The intact Giardia cyst wall is thin (,400 nm), easily fractured by sonication, and impermeable to small molecules. Curled fibrils of the GalNAc homopolymer are restricted to a narrow plane and are coated with linear arrays of oval-shaped protein complex. In contrast, cyst walls of Giardia treated with hot alkali to deproteinate fibrils of the GalNAc homopolymer are thick (,1.2 mm), resistant to sonication, and permeable. The deproteinated GalNAc homopolymer, which forms a loose lattice of curled fibrils, is bound by native CWP1 and CWP2, as well as by maltose-binding protein (MBP)-fusions containing the full-length CWP1 or CWP1 LRR. In contrast, neither MBP alone nor MBP fused to CWP1 CRR bind to the GalNAc homopolymer. Recombinant CWP1 binds to the GalNAc homopolymer within secretory vesicles of Giardia encysting in vitro. Fibrils of the GalNAc homopolymer are exposed during excystation or by treatment of heat-killed cysts with chymotrypsin, while deproteinated fibrils of the GalNAc homopolymer are degraded by extracts of Giardia cysts but not trophozoites. These results show the Leu-rich repeat domain of CWP1 is a lectin that binds to curled fibrils of the GalNAc homopolymer. During excystation, host and Giardia proteases appear to degrade bound CWPs, exposing fibrils of the GalNAc homopolymer that are digested by a stage-specific glycohydrolase.

Two cDNAs, isolated from a Trypanosoma cruzi amastigote library immunoscreened with sera from patients with Chagas disease, encode proteins with sequence homology to eukaryotic components of the cellular sorting and recycling machinery. These proteins, denominated TcAGL, present an N-terminal lectin domain and a C-terminal region containing repetitive amino acids and a poly-glutamine tract. They are products of polymorphic alleles of a single copy gene constitutively expressed during the parasite life cycle. Polyclonal antibodies obtained from mice immunized with the recombinant antigen recognize proteins with apparent molecular weight ranging from 95 to 120 kDa in cell lysates from all three life stages and in various strains of the parasite. Sera from Chagas disease patients recognize the recombinant antigen in ELISA and immunoprecipitation assays but not in Western blot assays under denaturing conditions. Consistent with its proposed role in the glycoprotein secreting pathway, immunofluorescence analyses and expression of a green fluorescent protein-tagged TcAGL protein indicate a sub-cellular localization in the vicinity of the flagellar pocket membrane and the Golgi complex of the parasite. Ó

Animal cells internalize specific extracellular macromolecules (ligands) by using specialized cell surface receptors that operate through a complex and highly regulated process known as receptor-mediated endocytosis, which involves the binding, internalization, and transfer of ligands through a series of distinct intracellular compartments. For the uptake of a variety of carbohydrate-containing macromolecules, such as glycoproteins, animal cells use specialized membrane-bound lectins as endocytic receptors that recognize different sugar residues or carbohydrate structures present on various ligands. The hepatic asialoglycoprotein receptor, which recognizes glycoconjugates containing terminal galactose or N-acetylgalactosamine residues, was the first membrane lectin discovered and has been a classical system for studying receptor-mediated endocytosis. Studies of how the asialoglycoprotein receptor functions have led to the discovery of two functionally distinct, parallel pathways of clathrin-mediated endocytosis (called the State 1 and State 2 pathways), which may also be utilized by all the other endocytic recycling receptor systems. Another endocytic membrane lectin, the hyaluronan/chondroitin sulfate receptor, which has recently been purified and cloned, is responsible for the turnover in mammals of these glycosaminoglycans, which are important components of extracellular matrices. We discuss the characteristics and physiological importance of these two proteins as examples of how lectins can function as endocytic receptors.

A highly purified trypsin inhibitor was obtained from Echinodorus paniculatus when an extract prepared from E. paniculatus seed flour (25 g l À1 , with 0.1 M ammonium acetate buffer, pH 8.3, under agitation for 6 min at 28°C) was chromatographed on Sephadex G-25 (12 ml h À1 ), followed by affinity chromatography on immobilized Cratylia mollis isolectins (Cra Iso 1,2,3-Sepharose). The column chromatography was performed at 24°C; the matrix was washed (30 ml h À1 ) with 0.1 M sodium phosphate buffer, pH 7.4 or with the same buffer containing 0.2 M glucose, followed by application of inhibitor sample and elution with 0.015 M sodium borate buffer, pH 7.4, or 1.0 M NaCl. A purified fraction of inhibitor was obtained by gel filtration chromatography (GF-450/HPLC column). Trypsin inhibitory activity was eliminated when the inhibitor was treated with metaperiodate showing that the carbohydrate moiety was important for trypsin inhibition. Binding of inhibitor was also evaluated on immobilized concanavalin A (Con A-Sepharose) using previously described chromatographic conditions with results similar to Cra Iso 1,2,3-Sepharose chromatography.

Griffonia simplicifolia, which reportedly binds to terminal N-acetylglucosamine residues in glycoconjugate oligosaccharides also shows affinity for glycogen. In the present study, the lectin was conjugated to horseradish peroxidase and applied to paraffin sections of kidney from streptozotocin-diabetic rats, insulintreated and untreated, and age-matched control rats. Griffonia simplicifolia agglutinin II detected glycogen in cortical ascending thick limbs of untreated diabetic rat kidneys as early as 24 h following injection of streptozotocin. The number of stained cells increased steadily so that by day 14 of diabetes the lectin reacted with nearly all of the cells lining ascending thick limbs in the cortex and adjacent outer stripe of the outer medulla. Glycogen was never identified in the inner medullary stripe. Comparison of Griffonia simplicifolia agglutinin II and periodic acid-Schiff staining revealed that periodic acid-Schiff could not clearly detect glycogen until 14 days following injection of streptozotocin, which substantiated earlier claims that Griffonia simplicifolia agglutinin II might be a more sensitive indicator of glycogen than periodic acid-Schiff. The distribution of glycoconjugate containing terminal N-acetylglucosamine stainable with the lectin was unchanged in diabetic kidneys. Griffonia simplicifolia agglutinin II served in the present study to further characterise the sequence of abnormal glycogen accumulation in streptozotocin-diabetic rat kidneys. In addition, it was shown that the lectin's ability to antedate periodic acid-Schiff detection of glycogen has utility in histochemical investigations in diabetes.

The present study evaluated the toxicity of Microgramma vacciniifolia rhizome lectin (MvRL) to Artemia salina, human tumour cell lines (larynx epidermoid carcinoma Hep-2, NCI-H292 lung mucoepidermoid carcinoma, and chronic myelocytic leukaemia K562), and normal peripheral blood mononuclear cells (PBMCs), as well as to Biomphalaria glabrata embryos and adults. MvRL was toxic to A. salina (LC 50 = 159.9 mg/mL), and exerted cytotoxic effects on NCI-H292 cells (IC 50 = 25.23 mg/mL). The lectin (1-100 mg/mL) did not affect the viability of K562 and Hep-2 tumour cells, as well as of PBMCs. MvRL concentration of 1, 10, and 100 mg/mL promoted malformations (mainly exogastrulation) in 7.8%, 22.5%, and 27.7% of embryos, respectively, as well as delayed embryo development in 42.0%, 69.5%, and 54.7% of embryos, respectively. MvRL at a concentration of 100 mg/mL killed B. glabrata embryos (17.7%) and adults (25%). Further, MvRL damaged B. glabrata reproductive processes, which was evidenced by observations that snails exposed to the lectin (100 mg/mL) deposited fewer eggs than those in the control group, and approximately 40% of the deposited eggs exhibited malformations. Comparison of these results with that from A. salina assay indicates that MvRL is adulticidal at the concentration range which is toxic to environment. In conclusion, the cytotoxicity of MvRL on tumour cell and absence of toxicity to normal cell indicate its potential as chemotherapeutic drug. Also, the study revealed that the lectin is able to promote deleterious effects on B. glabrata embryos at environmentally safe concentrations.

The human NK gene complex localized on chromosome 12p12.3-p13.2 codes for several lectin-like receptor genes expressed by NK cells as well as by other hematopoietic cells. In this study, by using the expressed sequence tag database we identified a novel receptor gene, designated as killer cell lectin-like receptor, subfamily F, member 1 (KLRF1), encoding a putative type II transmembrane glycoprotein. The KLRF1 gene has been localized on the high-resolution physical map of chromosome 12p. The genomic structure of the KLRF1 gene and the existence of one spliced variant are also described. KLRF1 was expressed at the mRNA level in peripheral blood leukocytes, activated NK cells, monocytes and NK and myeloid cell lines. The presence of two immunoreceptor tyrosine-based inhibitory-like motifs within the cytoplasmic tail of KLRF1 suggests an inhibitory role in NK cell and monocyte activity.

Differential scanning calorimetry was used to study the thermal unfolding of hevein, a 43-residue disulfide-rich protein whose three-dimensional structure has been determined by X-ray diffraction. In the range pH 2.0-3.7 this process was approximately 75 % reversible as judged by repeated scans on the same sample. The ratios of van't Hoff to calorimetric enthalpies were considerably larger than one, suggesting that intermolecular cooperation is involved in the unfolding of this protein. Alternatively, it is possible that the partial irreversibility of this process may cause distortions of the endotherm that affect the calculation of the van't Hoff enthalpy. Experimental changes in heat capacity and enthalpy were compared with those calculated from polar and nonpolar surface areas buried in the native state. It was found that when the unfolded state is represented as an extended chain without disulfide cross-links, experimental and calculated parameters agree well. However, if the unfolded protein is modeled with the presence of disulfide bridges, the agreement between the two sets of parameters is lost. The entropy changekesidue at 112 "C is considerably smaller than the average value for globular proteins, thus suggesting that, as expected, disulfide bonds strongly influence the entropy of the unfolded state of this protein. Fax: +52 5 7244666. Abbreviations. AH,,, calorimetric enthalpy ; AH,,, van't Hoff enthalpy ; AC,, change in heat capacity upon protein unfolding; T,, temperature at the maximum of the calorimetric curve; Av, change in the number of protons bound upon protein unfolding; A, and A,,, change in the solvent-accessible polar and apolar areas due to denaturation, respectively; AS,, unfolding entropy at T,.

I denne artikkelen diskuteres «blodtypedietten», som de siste årene har fått mye oppmerksomhet i Norge. Hovedhypotesen er at kostholdet bør tilpasses den enkeltes blodtype (ABO-systemet) for at vedkommende skal oppnå god helse. Selv om enkelte deler av det teoretiske fundamentet blodtypedietten bygger på er faglig veletablerte, er delene satt sammen til en resonnementkjede som ikke er holdbar. Ernaeringsmessig er blodtypedietten i det store og hele forsvarlig, med unntak av for lite kalsium i O-dietten og vitamin D i Adietten. De positive erfaringene som rapporteres er ikke vitenskapelig dokumentert, selv om det er fullt mulig å gjennomføre kontrollerte, randomiserte studier.

Background: Natural insecticides against the vector mosquito Aedes aegypti have been the object of research due to their high level of eco-safety. The water-soluble Moringa oleifera lectin (WSMoL) is a larvicidal agent against A. aegypti. This work reports the effects of WSMoL on oviposition and egg hatching of A. aegypti.

The article reviews the significant contributions to, and the present status of, applications of computational methods for the characterization and prediction of protein-carbohydrate interactions. After a presentation of the specific features of carbohydrate modeling, along with a brief description of the experimental data and general features of carbohydrate-protein interactions, the survey provides a thorough coverage of the available computational methods and tools. At the quantum-mechanical level, the use of both molecular orbitals and density-functional theory is critically assessed. These are followed by a presentation and critical evaluation of the applications of semiempirical and empirical methods: QM/MM, molecular dynamics, free-energy calculations, metadynamics, molecular robotics, and others. The usefulness of molecular docking in structural glycobiology is evaluated by considering recent docking- validation studies on a range of protein targets. The range of application...

Dietary lectins, present in beans and other edible plant products, pose a potential threat to consumers due to their capacity to induce histamine release from basophils. In this study, we analyzed the capacity of 16 common, in particular dietary, lectins to induce human basophils to secrete IL-4 and IL-13, the key promoters of Th2 responses and IgE synthesis. Several of the lectins, especially concanavalin A, lentil lectin, phytohemagglutinin, Pisum sativum agglutinin and Sambucus nigra agglutinin, triggered basophils to release IL-4 at concentrations of up to 1 ng/10 6 basophils. Lectins with high IL-4-inducing capacity also stimulated the release of IL-13 and histamine. Lectin-induced IL-4 and IL-13 release reached a maximum after 4-6 h and more than 18 h, respectively. Affinoblotting revealed that lectins with the capacity to induce mediator release bind to IgE, suggesting IgE binding as initial step of signal generation. In conclusion, several dietary lectins can trigger human basophils to release IL-4 and IL-13. Since lectins can enter the circulation after oral uptake, they might play a role in inducing the so-called early IL-4 required to switch the immune response towards a Th2 response and type I allergy. Abbreviations: Fc 4 RI: High-affinity IgE receptor IMDM: Iscove's modified Dulbecco's medium 918 H. Haas et al. Eur. J. Immunol. 1999. 29: 918-927

SRL is a cell wall associated developmental-stage specific lectin secreted by Sclerotium rolfsii, a soil-born pathogenic fungus. SRL displays specificity for TF antigen (Galβ1→3GalNAc-α-Ser//Thr) expressed in all cancer types and has tumour suppressing effects in vivo. Considering the immense potential of SRL in cancer research, we have generated two variant gene constructs of SRL and expressed in E. coli to refine the sugar specificity and solubility by altering the surface charge. SSR1 and SSR2 are two different recombinant variants of SRL, both of which recognize TF antigen but only SSR1 binds to

Peripherin is a type III intermediate filament (IF) abundantly expressed in developing neurons, but in the adult, it is primarily found in neurons extending to the peripheral nervous system. It has been suggested that peripherin may play a role in axonal elongation and/or cytoskeletal stabilization during development and regeneration. To further clarify the function of peripherin, we generated and characterized mice with a targeted disruption of the peripherin gene. The peripherin null mice were viable, reproduced normally and did not exhibit overt phenotypes. Microscopic analysis revealed no gross morphological defects in the ventral and dorsal roots, spinal cord, retina and gut, but protein analyses showed increased levels of the type IV IF a-internexin in ventral roots of peripherin null mice. Whereas the number and caliber of myelinated motor and sensory axons in the L5 roots remained unchanged in peripherin knockout mice, there was a substantial reduction ( 34%) in the number of L5 unmyelinated sensory fibers that correlated with a decreased binding of the lectin IB4. These results demonstrate a requirement of peripherin for the proper development of a subset of sensory neurons. Keywords: CGRP, dorsal root ganglion neurons, IB4, intermediate filament, sensory and motor neuronstransgenic mice.

This study examined the corticospinal tract in monkey by utilizing the anterograde transport of wheat germ lectin conjugated to horseradish peroxidase (WGA HRP) at the light microscopic level and the axonal transport of 3H-proteins with both light and electron microscopic autoradiographic techniques. The animals survived 3–9 days after the injections of 3H-leucine or 3H-leucine/WGA HRP into either motor or sensory cortices. With the laminar schema of Rexed as a guide to the layers of the spinal gray matter, qualitative and quantitative analyses of labeled projections of the corticospinal tract (CST) were made. With the light microscope, axons from the sensory cortex labeled with WGA-HRP could be observed in the contralateral spinal gray from lamina I to the border of laminae VI/VII, the heaviest distribution being located in medial III–VI. There was a small ipsilateral projection to V and VI. With 3H label, laminae I and II revealed few overlying silver grains; many grains overlay laminae III–VI. Projections from the motor cortex labeled with either WGA-HRP or 3H extended from the contralateral laminae III/IV border into the motor nucleus (lamina IX) and were seen to be somewhat more dense in the lateral areas of the spinal gray. The motor cortex projected heavily to ipsilateral VIII, and in sparse amounts to ipsilateral V and VI.Electron microscopy of radioactive axons from the sensory cortex to dorsal horn revealed many radioactive myelinated fibers and some labeled non-myelinated axons. Labeled terminals contacted medium to small dendrites; there were a few labeled C-type profiles in glomeruli and occasional axo-axonal or dendro-axonal contacts, the labeled cortical axons being the postsynaptic structure. In ventral horn following motor cortex injections, the labeled axons were all myelinated. The synaptic contacts were found on small, medium, and large proximal dendrites as well as on cell bodies. Labeled terminals which formed the central element in glomeruli were also seen in this region. Most of the labeled corticospinal terminals in dorsal and ventral horn contained rounded vesicles, but a significant number revealed pleomorphic vesicles. The relationship of these morphological findings to physiological studies of the CST is presented.

The mannose receptor (MR) is an endocytic protein on macrophages and dendritic cells, as well as on hepatic endothelial, kidney mesangial, tracheal smooth muscle, and retinal pigment epithelial cells. The extracellular portion contains two types of carbohydrate-recognition domain (CRD): eight membrane-proximal C-type CRDs and a membrane-distal cysteine-rich domain (Cys-MR). The former bind mannose-, N-acetylglucosamine-, and fucose-terminating oligosaccharides, and may be important in innate immunity towards microbial pathogens, and in antigen trapping for processing and presentation in adaptive immunity. Cys-MR binds to the sulfated carbohydrate chains of pituitary hormones and may have a role in hormonal clearance. A second feature of Cys-MR is binding to macrophages in marginal zones of the spleen, and to B cell areas in germinal centers which may help direct MR-bearing cells toward germinal centers during the immune response. Here we describe two novel classes of carbohydrate li...

Bone alkaline phosphatase is a marker of osteoblast activity. In order to study the posttranscriptional modification (glycosylation) of bone alkaline phosphatase in bone disease, we investigated the relationship between mass and catalytic activity of bone alkaline phosphatase in patients with osteoporosis and hyperthyroidism. Serum bone alkaline phosphatase aetivity was measured after lectin precipitation using the Iso-ALP test kit. Mass concentration of bone alkaline phosphatase was determined with an immunoradiometric assay (Tandem-R Ostase). In general, serum bone alkaline phosphatase mass and activity concentration correlated well. The activity : mass ratio of bone alkaline phosphatase was low in hyperthyroidism. Activation energy of the reaction catalysed by bone alkaline phosphatase was high in osteoporosis and in hyperthyroidism. Experiments with neuraminidase digestion further demonstrated that the thermodynamic heterogeneity of bone alkaline phosphatase can be explained by a different glycosylation of the enzyme.

In the postgenomic era, the analytical challenges associated with deciphering the structure–function relationship of complex oligosaccharides have resulted in the development of high-throughput technologies that are well suited for glycomic analysis. 1, 2 Lectin microarrays ...

The gene of the varicella-zoster virus (VZV) glycoprotein gE, engineered to code for a truncated molecule lacking the anchor and carboxy-terminal tail domains, was transfected into Chinese hamster ovary (CHO) cells via the pEEl4 mammalian expression vector. One recombinant cell line, CHO-gE-2-9, secreted high levels of truncated gE into the culture medium. The product was purified to near homogeneity by a combination of anion exchange, hydrophobic and metal-chelate chromatographies. Purified recombinant gE showed the expected amino-terminal sequence and its glycosylation pattern proved similar to that of the natural product. When injected into mice, using either Freund's or alum as adjuvant, the native truncated gE induced complement-dependent neutralizing antibodies. In contrast, when the molecule was first denatured, it lost immunogenicity with alum. These data show that the recombinant gE, although truncated, could potentially be included in a subunit vaccine against VZV infection.

This article reports the genotoxicity assessment of an extract of M. oleifera seed powder and the water-soluble Moringa oleifera lectin (WSMoL) isolated from seeds. The lectin isolated by chitin chromatography showed hemagglutinating activity with different erythrocytes, activity in a broad pH range (4.5 to 9.5), and retention of hemagglutinating activity after being heated to 100 • C. Genotoxicity of the seed extract and WSMoL were assessed using the cell-free plasmid DNA as well as the Salmonella typhimurium (Ames and Kado) assays with TA97, TA98, TA100, and TA102 in the presence or absence of hepatic metabolization. Seed extract at concentration (0.2 μg/μL) recommended to treat water was not genotoxic by Ames, Kado, and cell-free plasmid DNA assays. S. typhimurium strains showed to be sensitive to M. oleifera extract revealing a mutagenic effect at doses higher than 0.6 μg/μL with hepatic metabolization. The extract at doses higher than 0.4 μg/μL, without hepatic metabolization, was mutagenic for TA100 and TA102. WSMoL was nonmutagenic by used assays. The use of high concentrations of the extract may pose a risk to human health and the safe use of M. oleifera seed powder to treat water for human consumption requires more study; however, the purified lectin could be an alternative for water treatment.

Characterization of the lectins from onion (Allium cepa), shallot (A. ascalonicum) and leek (A. porrum) has shown that these lectins differ from previously isolated Alliaceae lectins not only in their molecular structure but also in their ability to inhibit retrovirus infection of target cells. cDNA libraries constructed from poly(A)-rich RNA isolated from young shoots of onion, shallot and leek were screened for lectin cDNA clones using colony hybridization. Sequence analysis of the lectin cDNA clones from these three species revealed a high degree of sequence similarity both at the nucleotide and at the amino acid level. Apparently the onion, shallot and leek lectins are translated from mRNAs of ca. 800 nucleotides. The primary translation products are preproproteins (ca. 19 kDa) which are converted into the mature lectin polypeptides (12.5-13 kDa) after post-translational modifications. Southern blot analysis of genomic DNA has shown that the lectins are most probably encoded by a family of closely related genes which is in good agreement with the sequence heterogeneity found between different lectin cDNA clones of one species.

We have shown that 4-Dibenzocyclooctynol (DIBO), which can easily be obtained by a streamlined synthetic approach, reacts exceptionally fast in the absence of a Cu I catalyst with azido-containing compounds to give stable triazoles. Chemical modifications of DIBO, such as oxidation of the alcohol to a ketone, increased the rate of strain promoted azide-alkyne cycloadditions (SPAAC). Installment of a ketone or oxime in the cyclooctyne ring resulted in fluorescent active compounds whereas this property was absent in the corresponding cycloaddition adducts, thereby providing the first example of a metal-free alkyne-azide fluoro-switch click reaction. The alcohol or ketone functions of the cyclooctynes offer a chemical handle to install a variety of different tags, thereby facilitating biological studies. It was found that DIBO modified with biotin combined with metabolic labeling with an azido-containing monosaccharide can determine relative quantities of sialic acid of living cells that have defects in glycosylation (Lec CHO cells). A combined use of metabolic labeling/SPAAC and lectin staining of cells that have defects in the Conserved Oligomeric Golgi (COG) complex revealed that such defects have a greater impact on O-glycan sialylation than galactosylation, whereas sialylation and galactosylation of N-glycans was similarly impacted. These results highlight that the fidelity of Golgi trafficking is a critical parameter for the types of oligosaccharides that are being biosynthesized by a cell. Furthermore, by modulating the quantity of biosynthesized sugar nucleotide, cells may have a means to selectively alter specific glycan structures of glycoproteins.

Aims: To assess the effects of two commonly used oral hypoglycemic medications metformin and pioglitazone on serum concentrations of omentin and leptin in patients with newly diagnosed type 2 diabetes. Methods: In a clinical trial setting (NCT01593371), patients were randomly allocated to either metformin 1000 mg daily (n=41), or pioglitazone 30 mg daily (n=50). Serum concentrations of omentin and leptin were measured at baseline and after 12 weeks. Patients' weight, waist circumference, blood pressure, fasting plasma glucose, fasting insulin, HbA1c, highly sensitive C-reactive protein, and serum lipids were also measured at the two visits. Results: Baseline concentrations of omentin and leptin were not different between the two arms of the trial. After three months, metformin decreased both omentin and leptin concentrations in women, and leptin concentrations only in men. On the other hand, pioglitazone reduced both adipokines only in women, but not men. Univariate and multivariate ANCOVA models revealed that both interventions are equally effective in reducing omentin concentration (p=0.497 for women and 0.344 for men in multivariate models controlling for the effects of confounding variables). Similarly, neither medication was more effective in reducing leptin concentrations after three months (p=0.822 for women and 0.441 for men in multivariate models). Conclusions: Metformin and pioglitazone at pharmacologic doses are equally effective in alteration of serum omentin and leptin concentrations in patients with diabetes, albeit sex differences in response to medications exist. Implication of these findings on long term management and complication prevention of diabetes needs to be elucidated.

Lectins was extracted from processed and unprocessed black-eyed peas, green beans, soybeans, peanuts, and red kidney beans by grinding using attrition mill and then soaked in distilled water. The proteins in the supernatant were precipitated by dissolving 10g of ammonium sulphate in 100ml of the supernatants. The lectin extracts were evaluated for hemagglutinin activity using human erythrocytes from blood group A + , B + , AB + , and O + , respectively. Result revealed that lectins from unprocessed soybean, black-eyed pea and green bean are blood group selective and may be classified as blood type specific lectins. While lectins from kidney beans and peanuts agglutinated all blood groups and may be classified as panhemagglutinin lectins, i.e. they agglutinated erythrocytes from all blood groups. Result shows that blood group O is less affected by hemagglutinin activity of lectin. Reduction/loss of hemagglutinin activity was observed with reduction in concentration of the crude lectin extracts in all unprocessed legumes. Boiling (at 100 0 C) for 30minutes completely inactivated hemagglutinin activity of lectin in Black eyed peas, soybeans, green beans and red kidney beans. Slight hemagglutinin activity was seen in lectin from roasted peanuts, indicating that dry heating may be very effective method to inactivate lectin completely. Also, in this study, lectins were extracted from rice (Oryza sativa), maize (Zea mays), sorghum (Sorghum bicolor), Acha (Digitaria excells), millet (Panicum maniceum); partial purification of the extract assayed from hemugglutinin activity using 4% of human erythrocyte in lectin buffer. Extract from these selected cereals showed positive and negative agglutination reaction with blood group A + , B + , O + and AB + depending on the concentration (dilution) and the blood group. Lectin extract from maize showed no agglutination reaction when diluted in blood group A + , B + , and AB + and even when processed and diluted. While crude lectin extract from rice showed no activity in blood group A + , B + , and AB + and when unprocessed and showed lectin agglutination activity when processed by boiling (cooking) showing that heat may encourage its activity. Blood group B + , and AB + showed no activity in all dilution. Also acha showed high agglutination activity in blood group A + , B + , AB + and O + but losses activity in dilution a 2 and a 3. Crude lectin extract from processed sorghum showed

The effector capacity of endogenous lectins on cell adhesion/growth prompts studies to turn them into pharmaceutically stable forms. Using human galectin-2 as a proof-of-principle model, we first introduced mutations at the site of one of the two Cys residues, that is, C57A, C57M, and C57S. Only the C57M variant was expressed in bacteria in soluble form in high yield. No notable aggregation of the modified homodimeric lectin occurred during 3 weeks of storage. This mutational process also facilitated the site-directed introduction of poly(ethylene glycol) into the remaining sulfhydryl group (Cys75). Product analysis revealed rather complete conjugation with one chain per subunit in the homodimer. We note that neither the secondary structure alteration nor the absence of binding ability to a glycoprotein (asialofetuin) was observed. The results thus document the feasibility of tailoring a human galectin for enhanced stability to aggregation as well as monoPEGylation, which enables further testing of biological properties including functionality as growth regulator and the rate of serum clearance.

The process of biofouling of marine structures and substrates, such as platforms or ship hulls, proceeds in multiple steps. Soon after the formation of an initial conditioning film, formed via the adsorption of organic particles to natural or manmade substrates, a population of different bacterial taxa associates under the formation of a biofilm. These microorganisms communicate through a complex quorum sensing network. Macro-foulers, e.g., barnacles, then settle and form a fouling layer on the marine surfaces, a process that globally has severe impacts both on the economy and on the environment. Since the ban of tributyltin, an efficient replacement of this antifouling compound by next-generation antifouling coatings that are environmentally more acceptable and also showing longer half-lives has not yet been developed. The sponges, as sessile filter-feeder animals, have evolved antifouling strategies to protect themselves against micro-and subsequent macrobiofouling processes. Experimental data are summarized and suggest that coating of the sponge surface with bio-silica contributes to the inhibition of the formation of a conditioning film. A direct adsorption of the surfaces by microorganisms can be impaired through poisoning the organisms with direct-acting secondary metabolites or toxic peptides. In addition, first, compounds from sponges have been identified that interfere with the anti-quorum sensing network. Sponge secondary metabolites acting selectively on diatom colonization have not yet been identified. Finally, it is outlined that direct-acting secondary metabolites inhibiting the growth of macro-fouling animals and those that poison the multidrug resistance pump are available. It is concluded that rational screening programs for inhibitors of the complex and dynamic problem of biofilm production, based on multidisciplinary studies and using sponges as a model, are required in the future.

S U M M A R Y The subcompartmentalization of the white pulp in the spleen is the result of interactions of specific resident stromal cells and migrating subtypes of lymphocytes. Because carbohydrate residues of cell membranes and extracellular matrices are involved in cell-cell and cell-matrix interactions, they were investigated in rat spleen by a broad panel of lectins. Splenic macrophages, which were also demonstrated by Perls' Prussian blue reaction, were labeled selectively by most mannose-specific lectins and gave the characteristic distribution patterns in all splenic (sub)compartments. One recently isolated lectin, Chelidonium majus agglutinin (CMA), visualized predominantly central arterioles, the reticular meshwork (RM) in the periarteriolar lymphatic sheaths (PALS), the circumferential reticulum cells limiting PALS and follicles, and some follicular dendritic cells (FDCs) in white pulp. The endothelial cells of venous sinuses in red pulp were also labeled by CMA and, if frozen sections were used, CMA also labeled the macrophages of the red pulp. Compared to CMA, the monoclonal antibody CD11, which can be used only in frozen sections, stained almost solely the fibrous (extracellular) component of the RM. Because CMA stains the reticulum cells in particular, it is better suited to visualize the stromal architecture of splenic white pulp than the monoclonal antibody. Because CMA can be applied to paraffin-embedded material, it is a particularly useful tool to study the splenic stromal architecture in archival material.

To meet the demands for food of the expanding world population, there is need of new ways for protecting plant crops against predators and pathogens while avoiding the use of environmentally aggressive chemicals. A milestone in this field was the introduction into crop plants of genes expressing Bacillus thuringiensis entomotoxic proteins. In spite of the success of this new technology, however, there are difficulties for acceptance of these 'anti-natural' products by the consumers and some concerns about its biosafety in mammals. An alternative could be exploring the plant's own defense mechanisms, by manipulating the expression of their endogenous defense proteins, or introducing an insect control gene derived from another plant. This review deals with the biochemical features and mechanisms of actions of plant proteins supposedly involved in defense mechanisms against insects, including lectins, ribosome-inactivating proteins, enzymes inhibitors, arcelins, chitinases, ureases, and modified storage proteins. The potentialities of genetic engineering of plants with increased resistance to insect predation relying on the repertoire of genes found in plants are also discussed. Several different genes encoding plant entomotoxic proteins have been introduced into crop genomes and many of these insect resistant plants are now being tested in field conditions or awaiting commercialization. q

A novel plant lectin was isolated from saltstressed rice (Oryza sativa L.) plants and partially characterized. The lectin occurs as a natural mixture of two closely related isoforms consisting of two identical non-covalently linked subunits of 15 kDa. Both isoforms are best inhibited by mannose and exhibit potent mitogenic activity towards T-lymphocytes. Biochemical analyses and sequence comparisons further revealed that the rice lectins belong to the subgroup of mannosebinding jacalin-related lectins. In addition, it could be demonstrated that the lectins described here correspond to the protein products of previously described saltstress-induced genes. Our results not only identify the rice lectin as a stress protein but also highlight the possible importance of protein-carbohydrate interactions in stress responses in plants. Abbreviations: Calsepa = Calystegia sepium agglutinin; ConA = concanavalin A; HCA = hydrophobic-cluster analysis; Heltuba = Helianthus tuberosus agglutinin; KM+ = mannose-binding lectin from Artocarpus integrifolia; MPA = Maclura pomifera agglutinin; Orysata = Oryza sativa agglutinin

Discrimination of self and nonself is one of the features of all animal species but the ways of elimination of nonself are different. Defense strategies of invertebrates, which lack antibodies and lymphocytes, are based on innate defense mechanisms. The study of such, undoubtedly less complex, defense mechanisms in invertebrates may shed,a new light on the more sophisticated immunity of vertebrates. The main aim of this review is to show on one experimental model -an oligochaete annelid -cellular and humoral defense pathways protecting against microbial infection.

Galectins are a family of galactose binding lectins that have become the focus of attention of cancer biologists due to their numerous regulatory roles in normal cellular metabolism and also because of their altered levels in various cancers. They are reportedly similar to several prominent and established modulators of apoptosis. In this review, we present a brief outline of the advancements in the methodology used to detect and identify them and their therapeutic applications in cancer. Their possible interactions with other glycoconjugates are also discussed and a vision for their future use in diagnosis and therapeutics is provided.

Projections from the nucleus tractus solitarii (NTS) to autonomic control regions of the ventrolateral medulla, particularly the nucleus reticularis rostroventrolateralis (RVL), which serves as a tonic vasomotor center, were analyzed in rat by anterograde, retrograde, and combined axonal transport techniques.