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Human melanoma antigen O-acetylated ganglioside GD3 is recognized by Cancer antennarius lectin.

A lectin that can specifically bind to O-acetylsialic acids, found in glycoproteins and gangliosides, was purified to homogeneity from a crab Cancer antennarius (crab lectin) (Ravindranath, M. H., Higa, H. H., Cooper, E. L., and Paulson, J. C. (1985) J. Biol. Chem. 260, 8850-8856; Correction (1986) J. Biol. Chem. 261, 1983; Ravindranath, M. H., and Paulson, J. C. (1987) Methods Enzymol. 138, 520-527). We tested lectin binding to human melanoma cell lines to identify O-acetylsialylated gangliosides on the melanoma cell surface. The highest degree of binding of the crab lectin was demonstrated on a melanoma cell line, UCLASO-M25. To confirm that the binding was due to O-acetylsialic acid in the alkali-labile gangliosides, the gangliosides were isolated and purified from M25 cells and individually coated onto sheep asialoerythrocytes, which served as targets in an agglutination assay using the lectin. The crab lectin agglutinated the asialo-sheep erythrocytes coated with alkali-labile gangliosides, but the lectin failed to agglutinate the asialoerythrocytes coated with GM3, GD3, and base-treated gangliosides. Subsequently, the purified alkali-labile M25 ganglioside was base-treated and applied to TLC, and we found that it was converted to a slower migrating species identical to the disialolactosylceramide (GD3). These results indicate that O-acetyl GD3 expressed on the melanoma cell surface is recognized by the lectin. Because O-acetyl GD3 is not expressed on human normal tissues, we examined the capability of O-acetyl GD3 to induce immune responses in man. Sera from patients with melanoma were tested against M25 cells in an immuneadherence assay, and those positive to the M25 cell line were further tested for specificity to O-acetyl gangliosides. The presence of autoantibodies to O-acetyl-GD3 in melanoma sera was confirmed by blocking of the antigen sites on M25 cells by the lectin or preabsorption of the sera with erythrocytes bearing O-acetyl gangliosides. The data provide evidence that O-acetyl-GD3 may represent an important tumor marker for detection and treatment of human melanoma.

Acetylation↗

Lack of specificity of brain gangliosides in the modulation of lymphocyte activation.

A potential role for glycolipid gangliosides to act as immunomodulating agents has been suggested. Most studies have employed brain gangliosides. We have systematically investigated highly purified murine brain gangliosides for their ability to modulate lymphocyte activation. All sialic acid classes of ganglioside inhibited lipopolysaccharide (LPS)-induced antibody secretion and all polysialated gangliosides inhibited LPS-induced DNA synthesis. Monosialated gangliosides had no effect on DNA synthesis induced by LPS. 8-BrcGMP-induced DNA synthesis was also inhibited, suggesting that a negative signal was delivered to B lymphocytes by co-cultivation with exogenous gangliosides. The lack of specificity with respect to sialic acid class observed in these studies suggests that further investigation of an immunomodulatory role for gangliosides focus on endogenous lymphocyte gangliosides.

Animals↗

Gangliosides and synaptic transmission.

Several physiologically relevant findings are reported indicating a functional involvement of gangliosides (sialoglycolipids) in the process of neuronal transmission: a) changes of gangliosides in both concentration and composition during developmental maturation of the CNS and with the level of phylogenetic organization; b) metabolic variations of gangliosides in relation to sensory stimulations; c) involvement of gangliosides in the process of thermal adaptations; d) correlations between long-lasting compensatory changes in the composition of brain gangliosides and long-term changes in the bio-electrical activity of the CNS and the learning ability; e) pronounced ability of gangliosides to complex with Ca2+-ions. On the basis of these experimental data an actualized hypothesis on the molecular interaction of brain gangliosides in the process of synaptic transmission is presented. The main feature of this hypothesis is the assumption that when the negatively charged sialic acids of gangliosides form complexes with Ca2+-ions (cluster-formation) these parts of the synaptic membrane become rigid. Dissociation of the Ca2+-ganglioside-complexes induces an "opening" of the presynaptic zone of interactions by means of an increase of membraneous fluidity, together with Ca2+-influx thus enabling fusion of vesicles and release of transmitter.

Acclimatization↗

Gangliosides enhance the membrane actions of ethanol and pentobarbital.

The physical properties of vesicles containing phosphatidylcholine (PC) and gangliosides were evaluated by the fluorescent probes 1,6-diphenyl-1,3,5-hexatriene, trans- parinarate (probes of the lower acyl regions), and 1-(4- trimethylammoniumphenyl )-6-phenyl-1,3,5-hexatriene (a probe of the glycerol backbone and upper acyl regions). Exposure of PC vesicles to ethanol produced a decrease in polarization of fluorescence of these probes, but large concentrations (200-600 mM) were required. Incorporation of 10 mole% rat brain synaptic gangliosides into the PC vesicles enhanced the effect of ethanol by several-fold. Synaptic gangliosides also enhanced the effect of ethanol on vesicles containing cholesterol in addition to PC. Comparison of mono-, di-, tri-, and asialogangliosides , sphingosine, and sphingomyelin indicated the importance of the oligosaccharide moiety, but not the sialic acid residues, in the enhancement of ethanol action by gangliosides. Variation of assay temperature and the acyl composition of the PC demonstrated that gangliosides increased the effects of ethanol only if the lipids were near the phase transition temperature. Gangliosides also produced a marked enhancement of the effect of pentobarbital on fluorescence polarization of probes in PC vesicles. The membrane partioning of pentobarbital was increased by gangliosides, but it was not clear that this increase in drug concentration in the membrane was sufficient to account for the large change in fluorescence polarization. These results suggest that ganglioside content may be an important determinant of the effects of alcohols and barbiturates on biomembranes. In particular, the high drug sensitivity of neuronal membranes may be related to the enrichment of gangliosides in these membranes.

Animals↗

Isolation and structural characterization of human lymphocyte and neutrophil gangliosides.

Gangliosides were isolated from purified preparations of human peripheral blood lymphocytes and neutrophils. Structural analyses and comparisons were performed by direct probe mass spectrometry and by degradation studies with the following enzymes: Escherichia freundii endo-beta-galactosidase; Clostridium perfringens and Arthrobacter ureafaciens neuraminidase; and jack bean beta-N-acetylhexosaminidase and beta-galactosidase. This combination of techniques allowed us to obtain carbohydrate composition and sequence information without the aid of methylation or carbohydrate compositional analyses using only 1-2 mg of purified gangliosides. On the basis of these studies we propose that human lymphocytes and neutrophils have gangliosides with the following structures. NeuAc alpha 2 leads to 3Gal beta 1 leads to 4Glc beta 1 leads to 1Cer Structure A NeuAc alpha 2 leads to ? GlcNAc beta 1 leads to 3Gal beta 1 leads to 4Glc beta 1 leads to 1Cer Structure B NeuAc alpha 2 leads to ? Gal beta 1 leads to 3,4GlcNAc beta 1 leads to 3Gal beta 1 leads to 4Glc beta 1 leads to 1Cer Structure C All three compounds were isolated from both cell types with structure A being the major lymphocyte ganglioside and structure C the major neutrophil ganglioside. Structure B is a novel ganglioside and may represent a leukocyte-specific glycosphingolipid. Neuraminidase degradation studies demonstrated that only one ganglioside species of each cell type contains an internally linked sialic acid residue, and on the basis of thin layer chromatographic analysis this component is the same as the major brain ganglioside, GM1 (II3-N-acetylneuraminosyl-gangliotetraosylceramide). In addition, large gangliosides with the general structure NeuAc alpha 2 leads to ?(Gal beta 1 leads to 3,4GlcNAc beta 1 leads to 3)n Gal beta 1 leads to 4Glc beta 1 leads to 1Cer were isolated. These results are discussed as they relate to blood group antigens and specific cell surface markers in human leukocytes.

Carbohydrate Conformation↗

Glycolipids of murine lymphocyte subpopulations. Structural characterization of thymus gangliosides.

Several sialic acid-containing glycolipids have been isolated and purified from murine thymus. Eleven of these acidic glycolipids (gangliosides) have been analyzed by several techniques, permitting us to propose a tentative structure for each. Sugar analysis reveals that all 11 gangliosides contain the same base structure, Gal(beta 1-3)GalNAc(beta 1-4)Gal(beta 1-4)Glc(beta 1-1) ceramide, which is gangliotetraosylceramide. No glucosamine or fucose is present in glycolipids from this tissue. There appears to be considerable heterogeneity in the sialic acid moiety of these gangliosides with respect to their structures and linkages. One ganglioside contains only N-glycolylneuraminic acid. Four of the gangliosides are common brain-type structures and contain sialic acid linked to internal galactose which is insusceptible to Vibrio cholerae neuraminidase. In addition, four of the gangliosides are susceptible to base hydrolysis and are believed to contain O-acetyl derivatives of sialic acid. Two gangliosides have also been shown to contain both N-acetyl- and N-glycolylneuraminic acid. There is also heterogeneity within the ceramide portion of some gangliosides which may be responsible for the appearance of an additional three ganglioside bands on thin layer chromatography.

Animals↗

Characterization and quantitative determination of gangliosides and neutral glycosphingolipids in human liver.

The neutral and acidic glycolipids from the liver of an 11-year-old male were quantitatively isolated and characterized. The total concentration of gangliosides was also determined in sampled from five other human livers. Lipid-bound sialic acid varied between 190-248 nmol/g with a mean value of 212 nmol/g. The major ganglioside was GM3, which represented 91.6% of the sialic acid. Besides GM3 and GM1, a wide variety of other minor monosialogangliosides were isolated. Gangliosides of the gangliotetraose series with up to four sialic acids were demonstrated in human liver for the first time. The composition of the ceramide portion of the gangliotetraose gangliosides was considerably different from that of the "visceral" gangliosides, GM3, GD3, and LM1 (sialosyl-lactoneotetraosylceramide), which suggests that these two groups of gangliosides are biosynthesized in two different pools. The concentration of the neutral glycolipids was approximately the same as that of the gangliosides. Lactosylceramide was the largest fraction, closely followed by galactosylceramide, glucosylceramide, and globotriaosylceramide. The ceramide composition of the neutral glycolipids resembled that of the "visceral" gangliosides, suggesting that they are metabolically related. 2-Hydroxy fatty acids were found in glucosyl-, galactosyl-, and lactosylceramides as well as in ganglioside GM3.

Adolescent↗

Gangliosides of human, cat, and rabbit spinal cords and cord myelin.

Gangliosides were isolated from whole spinal cords and cord myelin of human, cat, and rabbit by a revised methodology. The method included the sequential application of DEAE-Sephadex column chromatography, base treatment, Sephadex G-50 column chromatography, and finally Iatrobeads column chromatography. The human whole spinal cord was found to contain about one-tenth of the ganglioside concentration as in cerebral gray matter and about one-third of that in cerebral white matter. Low levels of gangliosides were also found in cat and rabbit whole cords. Only N-acetyl neuraminic acid could be detected in the ganglioside fractions of all three species. The whole cords also possessed unique ganglioside patterns when compared with the patterns of cerebral tissues. The most prominent and consistent features were the reduced concentration of Gd1a and increased amounts of Gm3 and Gd3. Human, but not cat and rabbit, spinal cord also contained Gm4 as one of the major gangliosides. Myelin prepared from the spinal cords of all three species also contained gangliosides. The amounts were only about half of those in the respective cerebral white matter myelin. The cord myelin ganglioside pattern was generally similar to the cerebral white matter myelin within the same species. Gm1 was the most abundant ganglioside in the cord myelin. Gm4 was found to be highly enriched only in myelin prepared from human sources.

Animals↗

Fluctuations in levels of various gangliosides in the different tracts of the oviduct of the frog (Rana esculenta) during the reproductive cycle.

The ganglioside content and pattern have been followed in the different tracts (rectus, convoluted and uterine) of the frog oviduct during the reproductive cycle. The main variations we observed are: a) average higher levels of ganglioside sialic acid in the preovulatory phase, with two peaks in March and April for the convoluted and rectus tract, respectively, and a more homogenous behaviour for the uterine tract; in all three tracts of the oviduct a minimum coincident with the ovulation has been found; b) a balanced presence of sulfolipids and gangliosides in the uterine tract: in fact sulfolipids, whose variations have been determined in a previous work, are higher when gangliosides are lower and vice versa, maintaining nearly constant the total negative charge due to these glycolipids; c) an alternate fluctuation of monosialo- and disialo-gangliosides in the preovulatory phase and a net trend toward the increase of monosialo- and the decrease of disialogangliosides in the postovulatory phase; trisialo-gangliosides are in general less represented and show less marked variations; d) the presence of particular gangliosides in particular moments of the reproductive cycle: Fuc-GM1, a fucosylated ganglioside, is higher than the more represented GM1 during the ovulation, while GD1 alpha, a ganglioside with a sialic acid residue linked to GalNAc, is steadily present in all three tracts after ovulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Immunohistochemical localization of ganglioside components in hepatocellular carcinoma and liver cirrhosis using monoclonal antibody.

BACKGROUND: In our previous studies (1, 2) a remarkable difference in the patterns of gangliosides was demonstrated between normal human liver and hepatocellular carcinoma (HCC) on thin layer chromatography, consisting of a marked increased in GM2 and the unidentified gangliosides. The proliferation of the unidentified gangliosides was also revealed in certain types of liver cirrhosis. The precise chemical structure of the unidentified components still remains to be elucidated, and there are no comprehensive data on these components investigated in liver tissue from patients with liver cirrhosis, HCC, or malignancies of the biliary and gastrointestinal tracts. In view of this, we studied the immunohistochemical localization of these ganglioside components in these tissues. EXPERIMENTAL DESIGN: Two unidentified gangliosides were extracted and monoclonal antibody were prepared. Localization of these gangliosides on tissue sections of liver obtained from HCC and liver cirrhosis, and malignancies of the biliary and gastrointestinal tracts was investigated by performing immunohistochemical staining using monoclonal antibody. The carbohydrate composition and sialic acid species of these components were determined by gas-liquid chromatography analysis. The sialic acid linkage to the terminal moiety of these components was also investigated by sialidase treatment. RESULTS: Staining with monoclonal antibody against specific gangliosides revealed dense brown granules in HCC tissue sections, whereas normal liver, kidney, and spleen sections were negative. Tissue sections of cirrhotic livers generally stained weakly except one case that showed slight to moderate staining. These antigens were immunohistochemically negligible in sections from one case each of gastric, colon, and common bile duct cancer. The ganglioside components identified in HCC were confirmed to be sialosylparagloboside with an alpha 2-6galactosyl terminus as determined by gas-liquid chromatography analysis and enzymatic hydrolysis with different sialidases. CONCLUSIONS: These components may be a useful marker in the detection of HCC and for subclassification of HCC from the standpoint of ganglioside metabolism.

Adult↗

Photochemical labeling of human erythrocyte membrane proteins with radioiodinated 4-azidosalicylic acid derivatives of G(M3), G(D3), G(M1), and FucG(M1) gangliosides.

Photoreactive gangliosides of high specific radioactivity may prove useful for studies on glycosphingolipid functions. We prepared 4-azidosalicylic acid (ASA) acylated derivatives of GM3, GD3, GM1, and FucGM1 gangliosides (gangliosides-ASA). Gangliosides-ASA were characterized by their TLC mobility, UV spectra, carbohydrate composition, and digestion with leech endoceramidase. After radioiodination to about 200 Ci/mmole gangliosides-ASA were used for photochemical labeling of human erythrocytes. Radioiodinated gangliosides-ASA were incorporated into erythrocytes in a time and concentration dependent manner, the kinetics and extent of incorporation being similar for all the gangliosides-ASA used. Radioiodinated gangliosides-ASA incorporated into erythrocytes were resistant to trypsin digestion while treatment with 1% BSA removed about 90% of the label. Incubation with cholera toxin protected radioiodinated GM1-ASA and, to a lesser extent, FucGM1-ASA but not GM3-ASA and GD3-ASA, against removal with BSA. After photolysis about 40-50% of radioactivity was firmly bound to erythrocyte lipids and proteins. The ratio of lipid- to protein-bound radioactivity ranged from 2.2:1 to 3.2:1. Photolabeled proteins were analyzed by SDS/PAGE followed by autoradiography. Band 3 was the most extensively photolabeled protein with all the radioiodinated gangliosides-ASA used. DIDS, an inhibitor of band 3 protein activity, caused reduction in photolabeling of this protein by about 20%.

Azides↗

Ganglioside expression in tissues of mice lacking beta2-microglobulin.

This study presents a comparative analysis of gangliosides from lymphoid (spleen and thymus) and other (brain, liver, lungs and muscle) tissues of C57BL/6 mice lacking the gene for beta2-microglobulin (beta2M), a constitutive component of the MHC class I molecule. Ganglioside fractions in the tissues of mice homozygous (beta2M-/-) and heterozygous (beta2M-/+) for the gene deletion were determined by high performance thin-layer chromatography (HPTLC), followed by immunostaining with specific polyclonal antibodies. Ubiquitous gangliosides GM3(Neu5Ac) and GM3(Neu5Gc) were the dominant gangliosides in the lungs of the control beta2M-/+ mice, whereas the homozygous knockout mice had substantially decreased expression of these structures. The lungs of the beta2M-/- mice also had reduced expression of T-lymphocyte-specific GM1b-type gangliosides (GM1b and GalNAc-GM1b). beta2M-deficient mice also had more GM1a and GD1a gangliosides in the liver, and several neolacto-series gangliosides were increased in the brain and lungs. This study provides in vivo evidence that the beta2M molecule can influence the acquisition of a distinct ganglioside assembly in different mouse organs, implicating its non-immunological functions.

Animals↗

[Gangliosides GM3 and GD3 in human stomach and breast tumors].

Gangliosides of human gastric and mammary tumours and of homologous normal tissues were studied by using biochemical methods and specific antisera. It was found that in most cases GM3, GD3 and GM1 are predominant gangliosides, whereas several polar components are minor ones. A comparison of the relative amount of ganglioside fractions revealed that in gastric tumours the per cent content of polar compounds is higher than in intact tissue; however, the absolute content of all gangliosides is markedly increased. A comparative study of the composition of mammary tumour and normal tissue gangliosides demonstrated two types of changes: i) the absolute content of all gangliosides in tumour tissue was increased and, ii) the increase in the content of total gangliosides was paralleled with the appearance of a new fraction (presumably GM4), the decrease of the GD3 content and the disappearance of polar gangliosides. A possible mechanism of this effect is discussed.

Breast Neoplasms↗

Coordinate regulation of ganglioside glycosyltransferases in differentiating NG108-15 neuroblastoma x glioma cells.

The enzymatic basis for ganglioside regulation during differentiation of NG108-15 mouse neuroblastoma x rat glioma hybrid cells was studied. This cell line contains four gangliosides that lie along the same biosynthetic pathway: GM3, GM2, GM1, and GD1a. Chemically induced neuronal differentiation of NG108-15 cells led to an 80% drop in the steady-state level of their major ganglioside, GM3, a sixfold increase in the level of a minor ganglioside, GM2 (which became the predominant ganglioside of differentiated cells); and relatively little change in the levels of GM1 and GD1a, which lie further along the same biosynthetic pathway. The enzymatic basis for this selective change in ganglioside expression was investigated by measuring the activity of two glycosyltransferases involved in ganglioside biosynthesis. UDP-N-acetylgalactosamine: GM3 N-acetylgalactosaminyltransferase (GM2-synthetase) activity increased fivefold during butyrate-induced differentiation, whereas UDP-galactose: GM2 galactosyltransferase (GM1-synthetase) activity decreased to 10% of its control level. Coordinate regulation of these two glycosyltransferases appears to be primarily responsible for the selective increase of GM2 expression during NG108-15 differentiation.

Alprostadil↗

An anti-ganglioside antibody-secreting hybridoma induces neuropathy in mice.

Immune responses against gangliosides are strongly implicated in the pathogenesis of some variants of Guillain-Barré syndrome (GBS). For example, IgG antibodies against GM1, GD1a, and related gangliosides are frequently present in patients with post-Campylobacter acute motor axonal neuropathy (AMAN) variant of GBS, and immunization of rabbits with GM1 has produced a model of AMAN. However, the role of anti-ganglioside antibodies in GBS continues to be debated because of lack of a passive transfer model. We recently have raised several monoclonal IgG anti-ganglioside antibodies. We passively transfer these antibodies by intraperitoneal hybridoma implantation and by systemic administration of purified anti-ganglioside antibodies in mice. Approximately half the animals implanted with an intraperitoneal clone of anti-ganglioside antibody-secreting hybridoma developed a patchy, predominantly axonal neuropathy affecting a small proportion of nerve fibers. In contrast to hybridoma implantation, passive transfer with systemically administered anti-ganglioside antibodies did not cause nerve fiber degeneration despite high titre circulating antibodies. Blood-nerve barrier studies indicate that animals implanted with hybridoma had leaky blood-nerve barrier compared to mice that received systemically administered anti-ganglioside antibodies. Our findings suggest that in addition to circulating antibodies, factors such as antibody accessibility and nerve fiber resistance to antibody-mediated injury play a role in the development of neuropathy.

Animals↗

Chemical residues of ganglioside molecules involved in interactions with lymphocyte surface targets leading to CD4 masking and inhibition of mitogenic proliferation.

Human lymphocyte CD4 becomes undetectable in the presence of exogenous gangliosides (CD4 masking), as originally described by Offner et al. (J. Immunol. 1987. 139: 3295). CD4 masking is apparently due to in situ rearrangement of the glycoprotein; since no direct binding of ganglioside to CD4 could be demonstrated, it was suggested that the effect could be mediated by interactions with other, as yet unidentified, surface structures. To gain insight into the structural requirements of the interaction(s) that leads to CD4 masking, we assayed the effects of a battery of gangliosides and of ganglioside derivatives on (a) CD4 masking; (b) cholera toxin binding (as a well known ganglioside-protein interaction) and (c) inhibition of lymphocyte mitogenic proliferation (as a second ganglioside interaction with a lymphocyte surface target). Our results indicate that the three interactions are distinctly different, since ganglioside chemical groups which are essential for one of the interactions are irrelevant for the others, and lead to the conclusion that gangliosides can interact with lymphocyte surface targets in a number of ways, causing a number of independent biological effects.

CD4 Antigens↗

Evaluation of the efficiency of an assay procedure for gangliosides in human serum.

An efficiency assessment of a ganglioside assay procedure was carried out on human serum gangliosides from healthy subjects of different sex and age. The analysis of the gangliosides, extracted with chloroform/methanol and purified by lipid partitioning, ion exchange column chromatographic separation and desalting procedures as described by Senn et al. (1989) Eur J Biochem 181: 657-62, was performed by HPTLC followed by densitometric quantification. The yield of the procedure, expressed as radioactivity recovery, was determined by adding GM3 ganglioside, tritium labelled at the sialic acid acetyl group and at the C3 position of sphingosine, to the lyophilized serum or by associating it with the serum lipoproteins. In spite of the fact that the extraction and purification procedures were performed exactly as described we found the radioactivity recovery to be variable (25-50%) and much lower than that proposed. Much of the radioactivity was found in the organic phase after lipid partitioning, whilst all the ganglioside purification steps led to some further loss. After the introduction of some modifications to the procedure the recovery improved, reaching 67-79%. The analyses on 33 samples of 5 ml showed a human serum ganglioside content of about 10 nmol ml-1 (as corrected for the recovery), and confirmed that GM3 ganglioside is the main component of the total serum ganglioside mixture.

Adult↗

Lactational changes in the fatty acid composition of human milk gangliosides.

The objectives of this work were to study the FA composition of milk gangliosides, as well as to gain further insight into the characterization of human milk gangliosides. The potential capacity of human milk gangliosides to adhere to human enterotoxigenic Escherichia coli (ETEC-strains) was also studied. Human milk gangliosides were isolated and identified by high-performance TLC or immunoassay. The latter also was used to assay bacterial adhesion. The FA composition of gangliosides was studied by GC. The presence of O-acetyl GD3 (Neu5,9Ac2alpha2-8 NeuAcalpha2-3Galbeta1-4GlcCer) and trace amounts of GM1 [Galgamma1]3-3GalNAcgamma1,-3(Neualpha2-3)Galbeta1-4GlcCerl in human milk was confirmed. Medium-chain FA were almost absent in colostrum, whereas in the subsequent stages they rose to 20%. The levels of long-chain FA decreased after colostrum. With respect to the degree of saturation, gangliosides from colostrum were richer in monounsaturated FA than gangliosides synthesized during the rest of the lactation period, opposite to the pattern for PUFA. A human-ETEC colonization factor antigen II-expressing strain showed binding capacity to human milk GM3 (NeuAcalpha2-3Gal[1-4GlcCer). New data on human milk gangliosides have been gathered. A thorough knowledge of their composition is needed since they may have important biological implications in regard to newborns' defense against infection.

Adhesiveness↗