Agglutination Process Research Articles

Penicillamine prevents ram sperm agglutination in media that support capacitation.

Ram spermatozoa are difficult to capacitate in vitro. Here we describe a further complication, the unreported phenomenon of head-to-head agglutination of ram spermatozoa following dilution in the capacitation medium Tyrodes plus albumin, lactate and pyruvate (TALP). Sperm agglutination is immediate, specific and persistent and is not associated with a loss of motility. Agglutination impedes in vitro sperm handling and analysis. So the objectives of this study were to investigate the cause of sperm agglutination and potential agents which may reduce agglutination. The percentage of non-agglutinated, motile spermatozoa increased when bicarbonate was omitted from complete TALP suggesting that bicarbonate ions stimulate the agglutination process. d-penicillamine (PEN), a nucleophilic thiol, was highly effective at reducing agglutination. The inclusion of 250 μM PEN in TALP reduced the incidence of motile, agglutinated spermatozoa from 76.7 ± 2.7% to 2.8 ± 1.4%. It was then assessed if PEN (1 mM) could be included in existing ram sperm capacitation protocols (TALP +1 mM dibutyryl cAMP, caffeine and theophylline) to produce spermatozoa that were simultaneously capacitated and non-agglutinated. This protocol resulted in a sperm population which displayed high levels of tyrosine phosphorylated proteins and lipid disordered membranes (merocyanine-540) while remaining motile, viable, acrosome-intact and non-agglutinated. In summary, PEN (1 mM) can be included in ram sperm capacitation protocols to reduce sperm agglutination and allow for the in vitro assessment of ram sperm capacitation.

Protein post-translational modification in host defense: the antimicrobial mechanism of action of human eosinophil cationic protein native forms.

Knowledge on the contribution of protein glycosylation in host defense antimicrobial peptides is still scarce. We have studied here how the post-translational modification pattern modulates the antimicrobial activity of one of the best characterized leukocyte granule proteins. The human eosinophil cationic protein (ECP), an eosinophil specific granule protein secreted during inflammation and infection, can target a wide variety of pathogens. Previous work in human eosinophil extracts identified several ECP native forms and glycosylation heterogeneity was found to contribute to the protein biological properties. In this study we analyze for the first time the antimicrobial activity of the distinct native proteins purified from healthy donor blood. Low and heavy molecular weight forms were tested on Escherichia coli cell cultures and compared with the recombinant non-glycosylated protein. Further analysis on model membranes provided an insight towards an understanding of the protein behavior at the cytoplasmic membrane level. The results highlight the significant reduction in protein toxicity and bacteria agglutination activity for heavy glycosylated fractions. Notwithstanding, the lower glycosylated fraction mostly retains the lipopolysaccharide binding affinity together with the cytoplasmic membrane depolarization and membrane leakage activities. From structural analysis we propose that heavy glycosylation interferes with the protein self-aggregation, hindering the cell agglutination and membrane disruption processes. The results suggest the contribution of post-translational modifications to the antimicrobial role of ECP in host defense.

English

This study investigated the feasibility and accuracy of blood types identification through deciduous teeth, using absorption-elution. A total of 15 deciduous teeth extracted from 15 children were used to identify blood types which were compared with the blood types of the children's parents.  The blood types of 5 (of 5) samples were fully-identified whose agglutination process took place less than a week. The blood types of four samples (of 5 samples) were identified if the agglutination process took place more than a week. The blood types of two samples (of 5 samples) were identified if the agglutination process took place more than a month. Taken together, our results suggest that deciduous teeth can be used to identify blood types using absorption-elution method for forensic odontology and the accuracy of blood types identification depends on the duration of agglutination.     Key words: Absorption-ellusi, blood type, deciduous teeth, agglutination.

An Alternative Chemical Redox Method for the Production of Bispecific Antibodies: Implication in Rapid Detection of Food Borne Pathogens

Bi-functional antibodies with the ability to bind two unrelated epitopes have remarkable potential in diagnostic and bio-sensing applications. In the present study, bispecific antibodies that recognize human red blood cell (RBC) and the food borne pathogen Listeria monocytogenes (L. monocytogenes) were engineered. The procedure involves initial reduction of a mixture of anti-RBC and anti-Listeria antibodies followed by gradual re-oxidation of the reduced disulphides. This facilitates association of the separated antibody chains and formation of hybrid immunoglobulins with affinity for the L. monocytogenes and human RBC. The bispecific antibodies caused the agglutination of the RBCs only in the presence of L. monocytogenes cells. The agglutination process necessitated the specific presence of L. monocytogenes and the red colored clumps formed were readily visible with naked eyes. The RBC agglutination assay described here provides a remarkably simple approach for the rapid and highly specific screening of various pathogens in their biological niches.

Monounsaturated Fatty Acid Ether Oligomers Formed during Heating of Virgin Olive Oil Show Agglutination Activity against Human Red Blood Cells

The present work focuses on the characterization of molecules formed when virgin olive oil is heated at 130 °C for 24 h open in air, which are found to be strong agglutinins. The hemagglutinating activity of the newly formed molecule isolated from the heated virgin olive oil sample was estimated against human red blood cells (RBCs). Dimers and polymers (high molecular weight molecules) were identified through thin layer chromatography (TLC) of the oil mixture. (1)H and (13)C nuclear magnetic resonance (NMR) and gas chromatography-mass spectroscopy (GC-MS) were the methods used for structural characterization. Among others, oligomerization of at least two monounsaturated fatty acids (FA) by an ether linkage between the hydrocarbon chains is involved. Light microscopy was used to characterize and visualize the agglutination process. Agglutination without fusion or lysis was observed. It was concluded that the heating of virgin olive oil open in air, among other effects, produces oligomerization as well as polymerization of unsaturated FA, possibly of monohydroxy, monounsaturated FA that is associated with strong hemagglutinating activity against human RBCs. The nutritional value and the effects on human health of such oligomers are not discussed in the literature and remain to be investigated.

Hemoglobin S Polymerization Effect on Water Self-Diffusion Coefficient

The transversal relaxation time, the effective transversal relaxation time and the water self-diffusion coefficient are evaluated during hemoglobin S polymerization. One homogeneous permanent magnet and one inhomogeneous and portable unilateral magnet with a very strong and constant static magnetic field gradient were utilized. The Carr-Purcell-Meiboom-Gill method was used before and after placing the studied samples 24 hours at 36°C to guarantee the polymerization. The transversal relaxation shows two exponents after polymerization supporting the concept of partially polymerized hemoglobin. The effective transversal relaxation time decreases around 40%, which can be explained by the increase of water self-diffusion coefficient 1.8 times as a main value. This result can be explained considering the effects of the agglutination process on the obstruction and hydration effects in a partially polymerized solution.

Application of a spectrally filtered probing light beam and RGB decomposition of microphotographs for flow registration of ultrasonically enhanced agglutination of erythrocytes

We propose a development of the flow microscopy method to increase the resolving power upon registration of erythrocyte agglutination. We experimentally show that the action of a ultrasonic standing wave on an agglutinating mixture blood-serum leads to the formation of so large erythrocytic immune complexes that it seems possible to propose a new two-wave optical method of registration of the process of erythrocyte agglutination using the RGB decomposition of microphotographs of the flow of the mixture under study. This approach increases the reliability of registration of erythrocyte agglutination and, consequently, increases the reliability of blood typing. Our results can be used in the development of instruments for automatic human blood typing.

Mass Transport Mechanism in the Retina of the Human Eye

The metabolic secretions produced by the cells inside the human eye are accumulated simultaneously in all of its parts; subject only to their origin. However, the symptomatology is well characterized by the location of accumulation. Genetic factors are directly related to how the clumps of metabolic secretions occur and the form of accumulation is very specific to the movement of the eyes based on the activities performed by the patient, so any outcome is unpredictable. The outer retinal layers are avascular and are supplied by diffusion from the choriocapillatis. Abnormalities of retinal blood circulation and drainage of metabolic secretions are key indicators to retinal dysfunction and may point to perturbations of systemic circulation. The weakening of the cornea bending mechanism results in the loss of variation of intraocular pressure. Consequently, due to the increase of its work activities, the weakening of the forced convection mechanism of the metabolic secretions produced in the retina initiates the accumulation of these secretions without uniformity in the retina, causing gradual obstruction of its system of mass transport, nutrients and metabolic secretions. This accumulation is readily observed by pressing ones hands for several minutes on closed eyes, subsequently, there is the perception of light in one without a region of uniformity. This work analyzes the anatomic structure and biophysics, depicted in the scientific literature, along with the symptoms of the presbyopia treatment of the first author of this paper, to understand that the accumulation of metabolic secretions causes an agglutination and dehydration process by decanting that triggers the loss of flexibility of the fibrous coat, vascular coat (uveal tissue) and nervous coat (retina) and consequently the gradual obstruction of the system of mass transport in the retina, compromising its nutrition and obstructing the drainage of its metabolic secretion.

Human blood group typing based on digital photographs of RBC agglutination process

A method for the monitoring of the human erythrocyte agglutination reaction in vitro, which is the basis for determining the human blood type (group), is proposed. The method is based on a statistical analysis of digital photographs of the agglutination process. It is experimentally shown that this analysis of photographs makes it possible to determine the probability that the agglutination reaction of erythrocytes of the studied specimen of blood with corresponding isohemagglutinating sera does occur. To increase the rate of the agglutination reaction of erythrocytes and to improve the sensitivity of the method of monitoring, the bioobject under study is subjected to the action of ultrasonic waves, as was previously proposed by the authors, and the result of the erythrocyte agglutination reaction is read optically. It is shown that, in principle, the method of statistical processing of digital photographs can be used to develop devices for automatic human blood typing in the AB0 and Rh systems.

Kinase‐Actuated Immunoaggregation of Peptide‐Conjugated Gold Nanoparticles

A simple and sensitive one-step approach for the rapid colorimetric detection of v-Src kinase using functionalized gold nanoparticles (NPs) in solution is reported. The generic concept operates without labeled ATP-derivatives, and signals are recorded directly by spectrophotometry. For the first time, surface plasmon resonance spectroscopy is applied to an NP system and compared to a semi-quantitative NP reaction model that was developed to determine the association rate constants and the rate-limiting step of the NP agglutination process.

Comparison of human RNase 3 and RNase 7 bactericidal action at the Gram‐negative and Gram‐positive bacterial cell wall

The eosinophil cationic protein/RNase 3 and the skin-derived RNase 7 are two human antimicrobial RNases involved in host innate immunity. Both belong to the RNase A superfamily and share a high cationicity and a common structural architecture. However, they present significant divergence at their primary structures, displaying either a high number of Arg or Lys residues, respectively. Previous comparative studies with a membrane model revealed two distinct mechanisms of action for lipid bilayer disruption. We have now compared their bactericidal activity, identifying some features that confer specificity at the bacterial cell wall level. RNase 3 displays a specific Escherichia coli cell agglutination activity, which is not shared by RNase 7. The RNase 3 agglutination process precedes the bacterial death and lysis event. In turn, RNase 7 can trigger the release of bacterial cell content without inducing any cell aggregation process. We hypothesize that the RNase 3 agglutination activity may depend on its high affinity for lipopolysaccharides and the presence of an N-terminal hydrophobic patch, and thus could facilitate host clearance activity at the infection focus by phagocytic cells. The present study suggests that the membrane disruption abilities do not solely explain the protein bacterial target preferences and highlights the key role of antimicrobial action at the bacterial cell wall level. An understanding of the interaction between antimicrobial proteins and their target at the bacterial envelope should aid in the design of alternative peptide-derived antibiotics.

The first microbialite - coral mounds in the Cenozoic (Uppermost Paleocene) from the Northern Tethys (Slovenia): Environmentally-triggered phase shifts preceding the PETM?

Upper Thanetian microbialite-coral mounds from the Adriatic Carbonate Platform (SW Slovenia) are described herein for the first time, representing an important case study of extensively microbially-cemented boundstones in the Early Paleogene. The mounds are constructed primarily by microbialites associated to small-sized coral colonies, forming metric bioconstructions in a mid-ramp setting. Detailed macroscopic and microscopic studies show that microbes are the major framework builders, playing a prominent role in the stabilization and growth of the mounds, with corals being the second most important component. Microbial carbonates represent up to 70% of the mounds, forming centimetric-thick crusts alternating with coral colonies. The microbial nature of the crusts is demonstrated by their growth form and internal microfabrics, showing accretionary, binding, and encrusting growth fabrics, often with gravity-defying geometries. Thin sections and polished slabs reveal a broad range of mesofabrics, with dense, structureless micrite (leiolite), laminated crusts (stromatolites), and clotted micritic masses (thrombolites). A first layer of micro-encrusters, including leiolites and thrombolites, occurs in cryptic habitats, whereas discontinuous stromatolites encrust the upper surface of corals. A second encrustation, the major mound construction phase, follows and is dominated by thrombolites, encrusting corals and other micro-encrusters. This sequence represents the basic constructional unit horizontally and vertically interlocked, in an irregular pattern, to form the mounds. The processes, which favored the deposition of these microbial carbonates, were mainly related to in situ precipitation, with minor evidences for grain agglutination and trapping processes. Scleractinian corals comprise moderately diversified community of small (centimetric) colonial, massive, platy encrusting, and branching forms. Coral colonies are distributed uniformly throughout the mounds without developing any ecological zonation. These features indicate that coral development remained at the pioneer stage throughout the mound growth. The spatial relationships between corals and microbialites, as well as the characteristics of microbial crusts and coral colonies, indicate a strong ecological competition between corals and microbes. A model for the evolution of the trophic structures during the mound growth is proposed, with changes in the paleoecology of the main bioconstructors triggered by frequent environmental perturbations. Turbidity and nutrient pressure, interpreted here as related to frequent recurrences of wet phases during the warm, humid climate of the Uppermost Thanetian, might have promoted temporary dominance of microbes over corals, causing rapid environmentally-driven “phase shifts” in the dominant biota.

Real-time monitoring of the cell agglutination process with a quartz crystal microbalance

The real-time monitoring of the agglutination process of human hepatic normal cells (L-02) at the quartz crystal microbalance (QCM) gold (Au) electrode was performed. Two lectins, concanavalin A (Con A) and wheat germ agglutinin (WGA), induced the cell agglutination, resulting in the different Δ f 0 and Δ R 1 responses from those caused by the normal cell attachment and growth. The cell–Con A–cell aggregates had higher affinity for the Au substrate due to the excellent adsorption ability of Con A, which was revealed by increased Δ f 0 and Δ R 1 shifts and the obvious mass effect of QCM. In contrast, the lower adsorption ability of cell–WGA–cell aggregates was related to the same characteristic of WGA, presenting the decreased Δ f 0 and Δ R 1 responses and the time-extended adhesion phase. Parallel microscopic observation experiments were also carried out and exhibited comparable results. The Δ f 0 responses during the processes of cell growth and cell agglutination were analyzed using the equations Δ f 0 = a 0 + a 1 e - t / τ 1 + a 2 e - t / τ 2 + a 3 e - t / τ 3 and Δ f 0 = a 0 + a 1 e - t / τ 1 + a 2 e - t / τ 2 , respectively. Furthermore, the current work proved that the QCM measurement technique based on cell agglutination was useful for discriminating hepatic normal cells (L-02) and hepatic cancer cells (Bel7402).

Development and evaluation of realistic microbioassays in freely suspended droplets on a chip

A novel technique for biomolecular detection in microliter droplets floating on the surface of high density oil is presented. Each droplet was captured and manipulated dielectrophoretically and was used as a site for a microscopic bioassay based on agglutination of antibody-conjugated particles. The results were read out by the pattern of unagglomerated gold nanoparticles collected on the droplet surface. Two formats of bioassays, namely gold only agglutination and gold and latex agglutination, were investigated experimentally by varying analyte concentration, particle size and concentration, number of antigen binding sites per particle, time for incubation, and rate of particle collection on the droplet surface. The microbioassays performance was also evaluated with ricin antibodies and compared to the ricin assays in field use. It is estimated that the droplet based assays require 100x smaller sample volume and are ten times more sensitive, though they require longer times to complete. The experiments were interpreted by modeling the kinetics of particle agglutination and mass transfer processes inside the droplets. The incubation time and antigen concentration values calculated by the model correlate well with the experimental results. The results could allow for development of efficient immunoassays on a chip requiring even smaller sample volumes.

Sugar−Quantum Dot Conjugates for a Selective and Sensitive Detection of Lectins

One-pot synthesized neoglycoconjugates with a reactive thiol group are introduced here for functionalization with carbohydrates for solubilization and stabilization of CdSe-ZnS quantum dots in aqueous solution. Three different sizes of quantum dots (QDs) with lactose, melibiose, and maltotriose on their surface have been utilized, for the first time, for lectin detection through agglutination assay. The sugar-QDs thus synthesized were characterized by transmission election microscopy (TEM), fluorescence, and absorption spectroscopy. Agglutination of sugar-QDs by three different lectins occurred through specific multivalent carbohydrate-lectin interactions and was studied extensively by monitoring the scattered light at 600 nm. This assay was very selective, which has been demonstrated by a more selective binding of soybean agglutinin (SBA) with melibiose-QD, as compared to lactose-QD, and specific deagglutination caused by alpha-d-galactose, while alpha-d-mannose did not show any effect. The detection sensitivity of the maltotriose-QD was tested with Concanavalin A (ConA), and as little as 100 nM of the lectin was detected using light scattering. The detection sensitivity of this protocol has been enhanced considerably by the fluorescence properties of QDs. This agglutination process seems to occur through formation of smaller soluble aggregates, which further associate to form larger aggregates.

Increased rate of apoptosis and diminished phagocytic ability of human neutrophils infected with Afa/Dr diffusely adhering Escherichia coli strains.

The proinflammatory effect of Afa/Dr diffusely adhering Escherichia coli (Afa/Dr DAEC) strains have been recently demonstrated in vitro by showing that polymorphonuclear leukocyte (PMN) transepithelial migration is induced after bacterial colonization of apical intestinal monolayers. The effect of Afa/Dr DAEC-PMN interaction on PMN behavior has been not investigated. Because of the putative virulence mechanism of PMN apoptosis during infectious diseases and taking into account the high level of expression of the decay-accelerating factor (DAF, or CD55), the receptor of Afa/Dr DAEC on PMNs, we sought to determine whether infection of PMNs by Afa/Dr DAEC strains could promote cell apoptosis. We looked at the behavior of PMNs incubated with Afa/Dr DAEC strains once they had transmigrated across polarized monolayers of intestinal (T84) cells. Infection of PMNs by Afa/Dr DAEC strains induced PMN apoptosis characterized by morphological nuclear changes, DNA fragmentation, caspase activation, and a high level of annexin V expression. However, transmigrated and nontransmigrated PMNs incubated with Afa/Dr DAEC strains showed similar elevated global caspase activities. PMN apoptosis depended on their agglutination, induced by Afa/Dr DAEC, and was still observed after preincubation of PMNs with anti-CD55 and/or anti-CD66 antibodies. Low levels of phagocytosis of Afa/Dr DAEC strains were observed both in nontransmigrated and in transmigrated PMNs compared to that observed with the control E. coli DH5alpha strain. Taken together, these data strongly suggest that interaction of Afa/Dr DAEC with PMNs may increase the bacterial virulence both by inducing PMN apoptosis through an agglutination process and by diminishing their phagocytic capacity.

85 Regeneration of protoplasts from disintegrated cells of the multi‐cellular marine green alga microdictyon umbilicatum

Protoplast regeneration from extruded cytoplasm of the multi‐cellular marine green alga Microdictyon umbilicatum (Velley) Zanardini (Cladophorales, Anadyomenaceae) was investigated. The early process of protoplast formation is comprised of two steps: agglutination of cell organelles into protoplasmic masses followed by generation of a temporary enclosing envelope around them. Agglutination of cell organelles was mediated by a lectin‐carbohydrate complementary system. Three sugars, D‐galactosamine, D‐glucosamine, and a‐D‐mannose, inhibited the agglutination process, and three complementary lectins for the above sugars, peanut agglutinin, Ricinus communis agglutinin and concanavalin A, bound to the surfaces of chloroplasts. Agglutination assay using human erythrocytes showed the presence of lectins specific for the above sugars in the algal vacuolar sap. The lectin has been purified by the use of D‐mannose agarose affinity column. Its Molecular weight was shown to be 36,000 dalton by SDS‐PAGE gel electrophoresis. When the basic regeneration process was accomplished, the cells chose one of two developmental strategies; about 70% of one‐celled protoplasts transformed into reproductive cells within two weeks after wounding, while others began cell division and grew into typical Microdictyon plants. Quadriflagellate swarmers were liberated from the reproductive cells, and they germinated into mature plants

A saga dos descendentes de rodínia na construção de Gondwana

All proposed reconstructions for the Rodinia supercontinent published during the last decade use to present a series of problems, of both fundaments (so including some of conceptual nature) and real geologic character. Besides a greater investment in the geological research as a whole, it seems to be necessary to give more attention to the already published articles and look to be acquainted with the regional geology of the countries of the third world. For the particular case of South America there are data enough to point out that the agglutination and the latter fission of Rodinia were rather long and diachronous tectonic processes, from a place to another. The singularity of Rodinia (if it have really existed) as a supercontinet was episodic, a short-living stage. With some good level of confidence, it is possible to state that some of the processes of agglutination (from ~1,5 up to 0,96 Ga) and fission (1,0 – 0,8-0,63 Ga) could coexist and compete for some time intervals, from a province to another.The number of blocks generated during the fission of Rodinia was large and diverse in terms of magnitude, crustal nature, thermal ages, as well as tectonic and rheological behavior during the agglutination of Gondwana, in Neoproterozoic times (up to the Ordovician, in Western Gondwana). Nevertheless, in all reconstrutions of Rodinia already published, only the three largest blocks have been considered, namely the Amazonico, S. Francisco-Congo and Rio de la Plata blocks, what is very far from the geological reality. This paper attempts to discriminate all descendant blocks of Rodinia and their roles and levels of crustal reworking during the construction/agglutination of Gondwana. A second step was to draw a preliminary comparative table, with all possible gradations, since the almost undisturbed large blocks (e.g. Amazonico) up to the small and entirely restructured blocks. There are some particular cases of strong reworking, where the discrimination of the previous blocks was only achieved with the progress of isotopic investigation.

Monounsaturated fatty acid oligomerization is responsible for the agglutination activity of heated virgin olive oil

The present study focuses on the isolation, purification and characterization of a molecule formed when virgin olive oil is heated at 130 °C for 24 h in air, that is found to be a strong agglutinin. The hemagglutinating activity of the novel molecule isolated from the heated olive oil was evaluated against human red blood cells (RBCs). Thin layer chromatography (TLC) of the oil mixture showed appearance of high molecular weight molecules, dimers and polymers. 13C nuclear magnetic resonance (NMR) and mass chromatography–mass spectroscopy (GC–MS) were used for structure elucidation. A linear oligomerization of monounsaturated fatty acids is involved. Light microscopy and electron microscopy were used to characterize and visualize the agglutination process. Agglutination without lysis or fusion was observed. The unheated olive oil and the isolated compound were also tested in-vitro against normal and malignant colon and breast cells. The results showed the highest reduction of tumor cells with the isolated novel compound. We conclude that virgin olive oil when heated in air produces oligomerization/polymerization of free unsaturated fatty acid possibly oleic acid (OA) that is a strong hemagglutinin against human RBCs with possible anti-cancer properties but with unknown nutritional effects on human health.

Macrocyclic glycoclusters. Self-aggregation and phosphate-induced agglutination behaviors of calix[4]resorcarene-based quadruple-chain amphiphiles with a huge oligosaccharide pool.

Macrocyclic glycocluster compounds 2n (n = 2-7) with four alkyl (undecyl) chains and eight oligosaccharide moieties on the opposite sides of the calix[4]resorcarene macrocycle are prepared from the reactions of the corresponding octaamine derivative with maltooligosaccharide lactones. Combined evidence from dynamic light scattering (DLS), gel permeation chromatography (GPC), and transmission electron microscopy (TEM) indicates that they form small micelle-like nanoparticles (d congruent with 3 nm) in water. In the presence of Na2HPO4/NaH2PO4, nanoparticles are agglutinated with phosphate ions as a glue to grow in size up to 60-100 nm, as revealed by DLS as well as microscopy (TEM and AFM). The phosphate-induced agglutination processes can be followed by surface plasmon resonance (SPR). Amphiphile 2n is readily immobilized on the hydrophobized sensor chip of SPR to give a closely packed monolayer with oligosaccharide moieties exposed to bulk water. While there is no further adsorption of 2n on the resulting monolayer, this does occur when the latter is pretreated with the phosphate salts, ultimately giving rise to a multilayer upon repeated treatment of the chip with 2n and Na2HPO4/NaH2PO4 in an alternate manner. Kinetic analyses show that the phosphate-mediated inter(saccharide) interactions in terms of rate and affinity are markedly dependent on the oligosaccharide chain lengths (n), becoming more favorable with increasing n's. The novel aggregation and agglutination behaviors observed are discussed in terms of immobilizable and irreversible micelles on the basis of the cone-shaped structure of quadruple-chain amphiphile 2n having a huge saccharide pool and the efficiency of multiple hydrogen bonding therein. The unique intermolecular binding properties of compound 22 and analogues so far reported are reviewed in light of the present finding.