Agglutinin Activity Research Articles

Influence of nutrition medium composition on biosynthesis of agglutinins of Rhizoctonia solani RS

The influence of the composition of the nutrient medium on the yield of biomass and the biosynthesis of agglutinins of the mold fungus strain Rhizoctonia solani RS was studied. At the first stage of research, the agglutinin-producing micromycete strain was grown on liquid nutrient media (potato-glucose (PG), Chapek, Sabouraud) at a temperature of 28°C for 8 days. At the next stage of research, the composition of the culture medium was modified in order to select sources of nitrogen (NH4NO3, (NH4)2SO4, NaNO3, arginine, tryptophan, asparagine, threonine, peptone) and carbon (sucrose, glucose, starch) nutrition. Cultivation of the Rh. solani RS on a liquid PG medium, containing glucose in an amount of 20.0 g/l led to a maximum yield of fungal biomass of 23.58 ± 1.30 g/l and the production of its agglutinins (titer 16384). The studied micromycete strain was characterized by the ability to actively use the amino acids threonine or asparagine when added to the above nutrient medium in an amount of 0.1 mg/ml. At the same time, the hemagglutination activity of the micromycete agglutinins increased twofold (titer 32768) compared to the activity of agglutinins on the PG medium without the addition of amino acids (titer 16384).

Co-extraction of soy protein and polysaccharide with lipid-lowering activity: Characterization of functional property, nutritional property and colonic fermentation property through a metabolomics approach

Defatted soy meal is a candidate for preparing lipid-lowering active ingredient. The study aimed to optimally co-extract soy protein and polysaccharide with lipid-lowering activity and improved functional and nutritional properties using non-conventional extraction techniques, i.e. thermal-, high pressure homogenization- and enzyme-assisted extraction. Non-conventional extraction enhanced the yields of soy protein and soluble saccharides and inactivated anti-nutrients. Soy extract (SE) prepared by pectinase-assisted extraction possessed not only strong in vitro lipid-lowering activity (i.e. cholesterol micellar inhibition activity, bile salts binding capacity, pancreatic lipase inhibition activity and cholesterol esterase inhibition activity), but also excellent functional (i.e. instant solubility of protein, low turbidity and acceptable color) and nutritional properties (i.e. none agglutinin activity). Resistant-digested fragment of 7S/11S played key role in lipid-lowering activity of SE. Moreover, SE beneficially altered colonic fermentation property of soy protein through promoting the release of bioactive peptides and organic acids, accelerating tryptophan and linoleic acid metabolism to produce indole acrylic acid, indole lactic acid and linoleic acid, and inhibiting putrefactive compounds (i.e. piperidine and tyramine). Oligosaccharide and polysaccharide in SE improved functional and nutritional properties of soy protein without compromising its lipid-lowering activity.

Mucin O-glycans suppress quorum-sensing pathways and genetic transformation in Streptococcus mutans.

INTRODUCTORY PARAGRAPHMucus barriers accommodate trillions of microbes throughout the human body while preventing pathogenic colonization.1 In the oral cavity, saliva containing the mucins MUC5B and MUC7 forms a pellicle that coats the soft tissue and teeth to prevent infection by oral pathogens, such as Streptococcus mutans.2 Salivary mucin can interact directly with microbes through selective agglutinin activity and bacterial binding,2–4 but the extent and basis of saliva’s protective functions are not well understood. Using an ex vivo saliva model, we identify MUC5B as an inhibitor of microbial virulence. Specifically, we find natively purified MUC5B downregulates the expression of quorum sensing pathways activated by the competence stimulating peptide (CSP) and the sigX inducing peptide (XIP).5 Further, MUC5B prevents the acquisition of antimicrobial resistance through natural genetic transformation, a process activated through quorum sensing. Our data reveal the effect of MUC5B is mediated by its associated glycans, which are potent suppressors of quorum sensing and genetic transformation, even when removed from the mucin backbone. Together, these results present mucin glycans as a host strategy for domesticating potentially pathogenic microbes without killing.

Oxidation induced by dielectric-barrier discharge (DBD) plasma treatment reduces soybean agglutinin activity

In this study, potential of dielectric-barrier discharge (DBD) plasma treatment (40 kV, 12 kHz at 1, 2, 3 and 4 min) to eliminate soybean agglutinin (SBA) activity was investigated in a SBA model system and soymilk. The plasma treatment decreased the SBA in the model system and hemagglutination activity was decreased by 87.31%. SDS-PAGE analysis confirmed the degradation of the SBA polypeptide chain. The multi-spectroscopic analysis revealed a two-stage structure alteration in the SBA upon exposure to the plasma treatment. Oxidation of NH-/NH2- at the peptide bond disrupted the hydrogen bonds and altered the secondary structure of SBA. Further oxidation of aromatic amino acid, cleavage of peptide bonds and the breakage of polypeptide led to the SBA fragmentation and complete unfolding of the protein. The SBA inactivation by the plasma treatment was confirmed in soymilk. Plasma treatment is a promising technology for the elimination of SBA in soybean product.

Purification and Partial Characterization of Agglutinin Lectin from Heamolymph of German Cockroach, Blattella germanica.

Background:Lectin molecules have crucial biological role in insects’ immune system. The aim of present study was to find the agglutinin activities in haemolymph of German cockroach, Belatella germanica with appropriate screening and purification.Methods:The heamolymph of cockroach was collected and agglutinin test performed against different animal and human red blood cells (RBC). Then sugar inhibition assay was carried out to find carbohydrate specific binding lectin. The proteins of haemolymph was purified using ion-exchange chromatography (HPLC) and each fraction was tested for agglutinin activity. Finally the molecular weight of the agglutinin protein was determined using SDS-page.Results:The most agglutinin activity of haemolymph was found against RBC of mouse at titer 1/128ml/L dilution and sugar inhibition assay showed that fucos, N-acetyglucoseamine and galactose reduced titer of agglutinin to ½ml/L. Only one fraction of heamolymph at rotation time of 36 minute showed agglutinin activity. The molecular weight of this lectin was measured as 120Kds.Conclusion:The range of agglutinin activities against different RBC indicates that the isolated lectin is not specific for a particular carbohydrate. In addition, the isolated lectin at low concentration present in heamolymph should be an innate lactin not secreted, because we found it without any trigger immunity of the insect.

Venom-dominated animal survival adaptation

Predation and defense are the core contents of ecological adaptation and physiological ecology of species. Animal venom system is a typical “biochemical weapon system” for animals to implement predation and defense functions, which is of great significance for understanding the physiological mechanism of animal ecological adaptation. Toxin molecules from various venomous animals have high structural and functional diversity. A variety of biological activities have been identified from the venom of poisonous animals, including neurotoxic activity, enzyme activity, cytotoxic activity, antibacterial activity, agglutinin activity, hemolytic activity, antithrombotic activity, coagulation activity, immunomodulatory activity, enzyme inhibitor activity, bradykinin enhancing activity, and antiviral activity. Toxins with high affinity and high selectivity are used by venomous animals to act on cell membranes, ion channels, receptors or enzymes to affect the nervous system, skeletal system, cardiovascular and cerebrovascular system and immune system, thus implementing fast and efficient predation and defense. In order to prey on poisonous animals, predators have been undergoing coevolution with poisonous animals. In this review, we introduce the molecular basis of predation and defense of venomous animals, the diversity of pharmacological functions and the adaptive mechanisms related to predation and defense of several typical venomous animals.

Development of Multiple Myeloma of the IgA Type in a Patient with Cold Agglutinin Disease: Transformation or Coincidence?

Cold agglutinin disease (CAD) is an autoimmune hemolytic anemia and a distinct, clonal bone marrow lymphoproliferative disorder, characterized in most cases by a monoclonal IgMκ serum protein. We describe a CAD patient presenting with a monoclonal immunoglobulin of the IgAλ class. For years, she remained asymptomatic apart from the hemolytic anemia until eventually she developed multiple myeloma (MM) of the IgAλ phenotype. Prior to the development of MM, her hemolytic anemia responded to rituximab monotherapy. After she was diagnosed with MM, both conditions responded well to bortezomib-based therapy. We performed further investigations to determine whether her MM represented a progression/transformation of CAD or an independent disease. Flow cytometry and biopsy findings convincingly confirmed two unrelated clonal B-cell disorders. On this background, we also discuss previously published reports on cold agglutinin activity in patients with IgA gammopathy. In conclusion, cold agglutinins of the IgA class do probably not result in CAD. If a monoclonal immunoglobulin other than IgMκ is found in a patient with CAD, the coexistence of two unrelated B-cell clones should be suspected.

Aktivitas aglutinin dari beberapa jenis rumput laut di Teluk Manado

Perairan Teluk Manado memiliki keragaman rumput laut yang tinggi. Penelitian ini bertujuan untuk mencari jenis-jenis rumput laut dari perairan Teluk Manado yang memiliki aktivitas aglutinin. Sampel rumput laut diambil di sekitar Teluk Manado. Setelah dibersihkan dan diidentifikasi jenisnya, sampel dihaluskan dan diekstraksi. Ekstrak diuji pada sel darah merah (eritrosit) manusia dari 4 golongan (A, B, O dan AB) dengan kontrol dibuat tanpa penambahan ekstrak. Aktivitas agglutinin ditentukan berdasarkan titer aglutinasi pada cekungan dengan pengenceran tertinggi yang masih menyebabkan aglutinasi sel. Aktivitas agglutinin tiap jenis rumput laut diuji pada berbagai temperatur dan pH. Dari lima jenis rumput laut yang diuji, hanya tiga yang mampu mengaglutinasi sel darah merah manusia golongan A, B, O dan AB. Halimeda makroloba dan H. taenicola tidak menunjukkan adanya aktivitas aglutinin. Berdasarkan hasil penelitian, dapat disimpulkan bahwa rumput laut jenis Ulva fasciata, Halymenia durvillaei dan Laurencia obtusa dari perairan teluk Manado memiliki aktivitas agglutinin.

EFFECT OF CHEMICAL, PHYSICAL AND ENZYMATIC TREATMENTS ON LACTOCOCCUS LACTIS' SURFACE AGGLUTININ AND ITS PRECIPITATING ABILITY

ABSTRACT The precipitating proterty of Lactococcus lactis agglutinin has a negative effect on yield of starch and expanded delaminating time during Chinese traditional mung bean starch processing; thus, it is of interest to find a way to destroy the precipitating property of the agglutinin. In the present study, L. lactis isolated from sour liquid was treated with chemical, physical, and enzymatic methods, respectively. It was found that 12.0–13.7 mol/L of ethanol or 12.3 mol/L of hydrogen peroxide or 0.1 mol/L of HCl, or 0.1 of mol/L NaOH or 0.3 mol/L trichloroacetic acid could destroy the ability of the agglutinin from L. lactis to precipitate starch and expand delaminating time significantly. Periodic acid sodium which destroys the structure of glycosyl also made L. lactis lose its agglutinin activity and precipitating ability. Similarly, pepsin and trypsin had the same function as periodic acid sodium did. The present study might provide a good way to increase the starch yield and delaminating time during the mung bean processing.PRACTICAL APPLICATIONSSour liquid processing has been used for hundreds of years to produce mung bean starch in China because this kind of starch had better quality than that from other processing methods. However, this processing consumed a lot of water while producing a large amount of waste water, resulting in environmental pollution.Many scientists considered that Lactococcus lactis in sour liquid is the main factor to precipitate starch. The agglutinin on the surface of L. lactis is directly involved in starch deposition. Therefore, our research focuses on improvement of starch yield by using L. lactis‐immobilized columns through which starch slurry passes. This technology will cut down waste water and meet the national requirements, so that we do not doubt the prospect of the technology.To develop this technology, we should learn what kinds of factors will affect the precipitating ability of agglutinin of L. lactis. This manuscript reports a study on how chemical, physical and enzymatic substance can affect the precipitating ability of agglutinin.

Immunoregulatory role for a public IgM idiotype in the induction of autoimmune diseases in Mycoplasma pneumoniae infection

Mycoplasma pneumoniae (MP) infection is associated with the emergence of various autoimmune disorders and autoantibody production. The most common autoantibodies induced are of anti-I specificity and express cold agglutinin (CA) activity. However, the mechanisms by which the microbial infection triggers the appearance of these autoantibodies are still unknown. To investigate these mechanisms, we used BALB/c mice as experimental models. In this paper, we show that BALB/c mice polyclonal antisera to MP react with human CA IgMs, and reciprocally, that BALB/c mice polyclonal antisera to human IgM CA react with MP. However, antibodies directed against MP and against CA IgM triggered by both immunizations represent two separate sets of antibodies. This was also confirmed using monoclonal antibodies derived from the immunized mice. Among these MAb we selected a monoclonal antibody MAb1D3 which reveals a cross-reactive idiotope (CRI) shared by human CA and other MIgMs with various autoantibody activities (anti-MAG and anti-IF). The CRI defined by MAb1D3 is a recurrent interspecies idiotope that is expressed by post infectious IgM antibodies to MP. Hence, we present in this study new data showing that the concomitant appearance of CAs and anti-MP IgM antibodies during acute MP infection is the consequence of a common idiotypic regulation of antibodies to infectious and to self antigens.

Extracellular protein production and morphogenesis of Lentinula edodes in submerged culture

Along with a brief review of Lentinula edodes (shiitake mushroom) submerged cultivation history within the framework of important extracellular proteins biosynthesis, this study contains the authors’ own results. The possibility of regulating the lectin activity of shiitake using the synthetic components is shown. The time course of lectin production in culture liquid of L. edodes in different media under submerged culture conditions was studied. The activity of agglutinins depended on the ratio between carbon and nitrogen sources and the pH of the culture medium. A relationship between the chemical composition of nutrient medium, the activity of extracellular lectins of L. edodes, and the formation of pigmented mycelial film in liquid culture has been found. The formulation of medium, on which the brown mycelial film appears in several days of submerged cultivation, is proposed. The results obtained make a contribution to the present notion of biochemical processes that give rise to the occurrence of the aforesaid morphological structure of shiitake. Finally, two extracellular lectins from the submerged culture of L. edodes have been isolated and purified to homogeneity. Their physicochemical properties and composition have been studied.

False-positive reactions of some monoclonal anti-c reagents

Cross-reactivities of monoclonal reagents for red blood cell (RBC) typing are seen very rarely, e.g. in some reagents for testing of ABO or Lewis antigens. We report on two patients in whom we observed weak reactions using a monoclonal anti-c reagent containing clone MS35, but no reactions with anti-c reagents containing other monoclonal or polyclonal antibodies. To resolve this discrepancy, we studied RHCE exon 1 (Ex1) and 2 from genomic DNA of one patient and performed quality controls of the false-positive reacting anti-c reagent. RHCE Ex1 showed no abnormalities as well as RH Ex2 examined with primers specific for Ex2 D/C. No amplification product of RH Ex2 was received with primers specific for c. A titration study with untreated, papain-treated and sialidase-treated adult and newborn RBCs showed anti-i reactivity of the false-positive reacting reagent. This reactivity could not be removed by absorption with c-negative newborn RBCs without reducing the anti-c titre in the same way, indicating a cross-reactivity. Some manufacturers give a remark on a possible cold agglutinin activity of their monoclonal anti-c reagents in the instruction leaflet, but in order to avoid such irritation, we recommend to remove this cross-reactivity by dilution during the manufacturing process.

Biochemical and Genomic Analysis of Neoculin Compared to Monocot Mannose-Binding Lectins

Neoculin occurring in an edible tropical fruit is a heterodimeric protein which has both sweetness and a taste-modifying activity that converts sourness to sweetness. Both the primary and the overall tertiary structures of neoculin resemble those of monocot mannose-binding lectins. This study investigated differences in biochemical properties between neoculin and the lectins. Structural comparison between the mannose-binding sites of lectins and the corresponding regions of neoculin showed that there is at least one amino acid substitution at each site in neoculin, suggesting a reason for the lack of its mannose-binding ability. This was consistent with hemagglutination assay data demonstrating that neoculin had no detectable agglutinin activity. DNA microarray analysis indicated that neoculin had no significant influence on gene expression in Caco-2 cell, whereas kidney bean lectin (Phaseolus vulgaris agglutinin) greatly influenced various gene expressions. These data strongly suggest that neoculin has no lectin-like properties, encouraging its practical use in the food industry.

Anti-D and anti-i activities are inseparable in V4-34-encoded monoclonal anti-D: the same framework 1 residues are required for both reactivities

The heavy-chain V4-34 germline gene segment is mandatory for pathologic cold-reacting autoantibodies with anti-I/i specificity (cold agglutinins) and is also preferentially used by monoclonal immunoglobulin M alloantibodies against D and other Rh antigens. The use of the V4-34 segment by monoclonal anti-D has previously been shown to also confer anti-I/i reactivity (cold agglutinin activity), which has implications for the use of such antibodies for Rh blood typing. V4-34 framework 1 (FR1) sequence is believed to be critical for cold agglutinin activity of cold agglutinins. The aim of this investigation was to use site-directed mutagenesis of a recombinant V4-34-encoded anti-D to determine the contribution of V4-34 FR1 sequence to anti-D activity and whether mutational modifications in the FR1 region could separately alter anti-D and anti-i activities. The results show that amino acid changes in V4-34 FR1 at W7, A23, and Y25 have a profound effect on anti-D activity as well as on anti-i activity. It was not possible to substantially reduce or remove anti-i activity without reducing anti-D activity to a comparable extent. The same nonpolar hydrophobic amino acids in FR1 are critical for maintaining both anti-D and anti-i activity. It is proposed that these residues influence the conformation of the antigen-binding site.

Anti‐D and anti‐i activities are inseparable in V4‐34‐encoded monoclonal anti‐D: the same framework 1 residues are required for both reactivities

BACKGROUND: The heavy‐chain V4‐34 germline gene segment is mandatory for pathologic cold‐reacting autoantibodies with anti‐I/i specificity (cold agglutinins) and is also preferentially used by monoclonal immunoglobulin M alloantibodies against D and other Rh antigens. The use of the V4‐34 segment by monoclonal anti‐D has previously been shown to also confer anti‐I/i reactivity (cold agglutinin activity), which has implications for the use of such antibodies for Rh blood typing. V4‐34 framework 1 (FR1) sequence is believed to be critical for cold agglutinin activity of cold agglutinins.STUDY DESIGN AND METHODS: The aim of this investigation was to use site‐directed mutagenesis of a recombinant V4‐34‐encoded anti‐D to determine the contribution of V4‐34 FR1 sequence to anti‐D activity and whether mutational modifications in the FR1 region could separately alter anti‐D and anti‐i activities.RESULTS: The results show that amino acid changes in V4‐34 FR1 at W7, A23, and Y25 have a profound effect on anti‐D activity as well as on anti‐i activity. It was not possible to substantially reduce or remove anti‐i activity without reducing anti‐D activity to a comparable extent.CONCLUSIONS: The same nonpolar hydrophobic amino acids in FR1 are critical for maintaining both anti‐D and anti‐i activity. It is proposed that these residues influence the conformation of the antigen‐binding site.

Deposition-associated diseases related with a monoclonal compound

Up to 3% of adults over 50 years of age show a monoclonal peak values in blood or urine. Findings and prognosis will be distinct in view of the nature of this factor. In B-cell neoplasias (multiple myeloma, Waldeström macroglobulinaemia, chronic myeloid leukaemia and non-Hodgkin lymphoma) the clinical pattern is dominated by the systemic effects produced by the expansion of the malign clone; the monoclonal protein may result in hyperviscosity syndrome or renal damage. On the other hand, there are other less frequent processes called diseases associated to monoclonal components, where the main clinical manifestations and prognosis depend of the biological effects of the monoclonal protein. With reference to this last group, which is the objective of this revision, no bone lesions, anaemia or a greater tendency to infections usually occur when compared with the first group. Even so, there are some cases of interposition between both groups: for instance, type IgM immunoglobulin present in Waldeström macroglobulinaemia may have cold agglutinin activity, and in the case of multiple myeloma, the clone may secrete amyloidogenic light chains.

Effect of culture medium composition on the activity of extracellular lectins of Lentinus edodes

The time course of lectin production in culture liquid of the basidial fungus Lentinus edodes strain F-249 in different media under the conditions of submerged culture was studied. The activity of agglutinins depended on the ratio between carbon and nitrogen sources and pH of culture medium. The activity of lectin in culture medium was maximal when the fungus was grown in a medium containing L-arabinose as a source of carbon and L-asparagine as a source of nitrogen (C : N ratio, (9.5-12): 1)) on the day 15-18 of culturing at pH 8-9.

On the Possible Function of Telfairia occidentalis Agglutinin in the Plant

The fate of Telfairia occidentalis seed Agglutinin, TOA, has been monitored during germination. While the level of the Agglutinin in the cotyledons decreased sharply in the first three days to about half of the initial level, it stabilises at this level for the following twelve days. In this interval, Agglutinin activity becomes manifest in the radicle on the fourth day, peaking on the fifth and decreasing rapidly thereafter. In the plumule, the lectin activity becomes manifest on the sixth day, peaks on the seventh and decreases rapidly thereafter. No lectin activity is detectable in any plant tissue including the rump of the cotyledons twenty-seven days after germination. The implications of these observations on the possible role of lectins in plants are discussed.

The Effect of the Carbohydrate Components of Pea Roots on the Enzymatic Activity of the Surface Agglutinins of Rhizobium leguminosarum bv. viciae 252

The interaction of the surface agglutinins of Rhizobium leguminosarum by. viciae 252 with the carbohydrate components of host pea roots alters the beta-glucosidase and proteolytic activities of the agglutinins.

Rapid cytotoxicity of human B lymphocytes induced by VH4-34 (VH4.21) gene-encoded monoclonal antibodies, II.

We have previously described complement-independent killing of human B lymphocytes by two IgM MoAbs derived from the VH4-34 (VH4.21) gene. Analysis of 17 independently derived VH4-34-encoded MoAbs shows that B cell toxicity is not limited to the two described MoAbs, but is a general property shared by a subset of MoAbs derived from the VH4-34 gene. As observed by two independent microscopy techniques, giant membrane pores were formed on target B cells within 10-15 min of exposure to cytotoxic VH4-34-derived MoAbs. Toxicity by individual MoAb correlated directly to its B cell binding intensity measured by FACS, i.e. stronger the binding greater the killing. Sequence analysis showed that V(H) region in germ-line or in near germ-line configuration was necessary but not sufficient for B cell binding. In addition, a particular sequence motif enriched in basic amino acids in the CDR3 may be required to supplement the reactivity mediated by the V(H) region of the MoAb molecule. VH4-34-encoded antibodies that fulfil the above sequence requirements have cold agglutinin activity towards the i antigen of cord erythrocytes. In vivo, such anti-i/anti-B cell antibodies are rarely detected in healthy adults, but serum levels are dramatically elevated in selective pathological conditions, such as systemic lupus erythematosus and infectious mononucleosis. This strict regulation may be related to the novel and rapid mechanism of human B cell toxicity demonstrated by antibodies encoded by a single human V(H) gene.