No abstract

Epithelial Ovarian cancer has the highest mortality rate among gynaecological cancers. Altered glycosylation is associated with oncogenic transformation producing tumor-associated carbohydrate antigens. We investigated the potential of natural occurring anti-glycan antibodies in the diagnosis of ovarian cancer by using printed glycan array. Anti-glycan antibodies bound to 203 chemically synthesized printed glycans were detected via biotin-streptavidin fluorescence system in serum of women with normal operative findings (healthy controls; n=24) and non-mucinous borderline or ovarian cancer of various FIGO stages (n=33). Data were validated measuring blood group associated di-, tri and tetra- saccharide antigens on known ABO blood groups. Anti-glycan antibodies demonstrated high reproducibility (rc>0.9). Cluster analysis identified repetitive patterns of specific core carbohydrate structures: 11 N-linked glycans, 3 O-linked glycans, 2 glycosphingolipids. Biomarker detection revealed 24 glycans including P1 (Galα1-4Galβ1-4GlcNAcβ; p<0.001) significantly discriminating between (low-) malignant tumors and healthy controls. Comparable sensitivity and specificity with tumor marker CA125 was achieved by a panel of multivariate selected and linear combined anti-glycan antibody signals (79.2% and 84.8%, respectively). Our findings demonstrate the potential of glycan arrays in the development of a new generation of biomarkers for ovarian cancer.

In common marmoset monkeys (Callithrix jacchus, order Primates), infants aged 2-28 days were deprived of parental care for 30-120 min/day in order to investigate the long-term effects of this neglect-stress model on affect and cognition in a primate species. Basal morning levels of urinary cortisol across the first year of life were unaffected in early deprived marmosets relative to their sibling controls. Basal morning levels of urinary dopamine were chronically increased. This peripheral increase in dopamine activity could represent a marker for central dopamine hyperactivity. Certainly, subadult early deprived marmosets exhibited performance deficits in two dopamine-regulated neuropsychological tasks. They demonstrated: (1) impaired behavioral inhibition in an object reaching with detour task, exhibiting significantly more nonreinforced forward reaches to a reward visible inside a cube that could only be retrieved through an opening to the side of the cube; and (2) impaired reversal learning in a two-way discrimination task based on visual icons presented on a touch-sensitive computer screen. These findings provide further evidence for the relevance of this novel primate model of parent-infant neglect to the environmental causes and mechanisms of human developmental psychopathology.

The mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) are nuclear transcription factors that mediate many of the basal and stress functions and effects of the corticosteroid hormones, including those related to brain development. Despite this, relatively little is known about the postnatal ontogeny of MR and GR gene and protein expression in the central nervous system, and this is particularly true of the primates, including humans. Here we describe the postnatal ontogeny of central MR and GR gene and protein expression in the common marmoset monkey. By developing marmoset-specific riboprobes and using in situ hybridization, it was demonstrated that MR mRNA expression in the dentate gyrus and Ammon's horn was significantly greater in marmoset infants (aged 4-6 weeks) than in neonates (1-2 days), juveniles (4-5 months) and adults (3-6 years), with expression in the latter three ontogenetic stages being broadly similar. In the same subjects and ontogenetic stages, GR mRNA expression was developmentally consistent in the marmoset dentate gyrus and Ammon's horn, as well as in the paraventricular nucleus of the hypothalamus. Qualitative immunohistochemical comparison of infants and adults demonstrated that MR protein expression in the hippocampus was, as for mRNA, also greater in infants than adults, and that hippocampal GR protein was, as for mRNA, also similar in infants and adults. The increase in MR mRNA expression between the stages of neonate and infant co-occurred with a reduction in basal plasma ACTH and cortisol titres. The ontogenetic profiles of MR and GR gene expression in the marmoset monkey are therefore fundamentally different from those described for the rat and the mouse. This evidence for the postnatal ontogeny of central corticosteroid nuclear receptor expression in a primate is important for understanding both the developmental stage-specific significance of stress exposure and its potential long-term effects on health and disease.

Glycan-binding antibodies form a significant subpopulation of both natural and acquired antibodies and play an important role in various immune processes. They are for example involved in innate immune responses, cancer, autoimmune diseases, and neurological disorders. In the present study, a microsphere-based flow-cytometric immunoassay (suspension array) was applied for multiplexed detection of glycan-binding antibodies in human serum. Several approaches for immobilization of glycoconjugates onto commercially available fluorescent microspheres were compared, and as the result, the design based on coupling of end-biotinylated glycopolymers has been selected. This method requires only minute amounts of glycans, similar to a printed glycan microarray. The resulting glyco-microspheres were used for detection of IgM and IgG antibodies directed against ABO blood group antigens. The possibility of multiplexing this assay was demonstrated with mixtures of microspheres modified with six different ABO related glycans. Multiplexed detection of anti-glycan IgM and IgG correlated well with singleplex assays (Pearson's correlation coefficient r = 0.95-0.99 for sera of different blood groups). The suspension array in singleplex format for A/B trisaccharide, H(di) and Le(x) microspheres corresponded well to the standard ELISA (r > 0.94). Therefore, the described method is promising for rapid, sensitive, and reproducible detection of anti-glycan antibodies in a multiplexed format.