ABSTRACT
Glycans mediate various biological processes through carbohydrate-protein interactions, and glycan microarrays have become indispensable tools for understanding these mechanisms. However, advances in functional glycomics are hindered by the absence of convenient and universal methods for obtaining natural glycan libraries with diverse structures from glycoconjugates. To address this challenge, we have developed an integrative approach that enables one-pot release and simultaneously capture, separation, structural characterization, and functional analysis of N/O-glycans. Using this approach, glycoconjugates are incubated with a pyrazolone-type heterobifunctional tag-ANPMP to obtain glycan-2ANPMP conjugates, which are then converted to glycan-AEPMP conjugates. We prepared a tagged glycan library from porcine gastric mucin, soy protein, human milk oligosaccharides, etc. Following derivatization by N-acetylation and permethylation, glycans were subjected to detailed structural characterization by ESI-MSn analysis, which revealed >83 highly pure glycan-AEPMPs containing various natural glycan epitopes. A shotgun microarray is constructed to study the fine details of glycan-bindings by proteins and antisera.
Subject(s)
Proteins , Pyrazolones , Animals , Humans , Swine , Glycoconjugates , Polysaccharides/chemistry , Glycomics/methodsABSTRACT
Glycans have been proven to play special roles in keeping human health as a class of nutritional and bioactive ingredients in many food materials. However, their broad use in the food industry is hindered by the lack of comprehensive analytical methods for high-quality food glycomics studies and large-quantity raw materials for their production. This study focuses on structural identification and quantitative comparison of bioactive N-glycans in seven species of livestock and poultry plasma as potential natural glycan resources by a novel comprehensive relative quantification strategy based on stable isotope labeling with nondeuterated and deuterated 4-methyl-1-(2-hydrazino-2-oxoethyl)-pyridinium bromide (d0/d7-HMP) in combination with linkage-specific derivatization of sialic acid residues. Methodological validation of the method in terms of detection sensitivity, signal resolution, quantification linearity, precision, and accuracy on model neutral and complicated sialylated glycans demonstrated its advantages over the existing methods. Based on this method, a series of bioactive N-glycans were found in seven species of livestock and poultry plasma, and their differences in structure, abundance percentages, and relative contents of N-glycans were revealed, demonstrating their excellent applicability for comprehensive food glycomics analysis and great exploitation potential of these plasma samples as large-quantity raw materials in producing bioactive N-glycans for application in food and pharmaceutical industries.
Subject(s)
Livestock , Poultry , Animals , Humans , Polysaccharides/chemistry , N-Acetylneuraminic Acid , Glycomics/methodsABSTRACT
Glycomics analysis has been undermined by the lack of structurally defined individual glycans as model compounds. However, it is challenging to prepare individual glycans from natural resources, mainly due to separation difficulties caused by highly diverse structure, complicated mixture form and chromophore-free property of naturally-existing glycans. In this study, we report a simple, universal and low-cost glycan separation strategy, glycoselection, which allows preparation of individual reducing glycans from their mixtures through reversible chromogenic derivatization by hydrazide chemistry in combination with two-dimensional high-performance liquid chromatography (2D-HPLC). Investigations on reaction conditions using lactose and maltodextrin as model glycans showed the feasibility of reversible hydrazide labeling and one-pot hydrazone conversion under mild conditions, the good stability of hydrazone-form derivatives of glycans in solution and the difference among seven selected hydrazine-carrying chromogenic reagents in product yields during glycan labeling and post-column detagging. The 2D-HPLC separation conditions were established on maltodextrin, from which fourteen highly-purified individual reducing oligo-glucans were ultimately obtained. Using this strategy, we also successfully prepared and identified eleven individual neutral reducing N-glycans from chicken ovalbumin and thirteen individual neutral reducing oligosaccharides and eight individual sialylated ones from human milk, demonstrating its good applicability to different types of reducing glycans as well as biological samples. Given the compatibility of individual reducing glycans with almost all of glycan derivatization protocols and analytical techniques of glycans and the potential of the method for larger scale application, this work provides a universal approach to compound-specific analysis of natural glycans and has great significance for glycomics studies.
Subject(s)
Glycomics , Hydrazones , Humans , Chromatography, High Pressure Liquid , Glycomics/methods , Polysaccharides/chemistry , HydrazinesABSTRACT
OBJECTIVE: Evidence has shown the importance of tumor necrosis factor alpha (TNF-alpha) in the pathophysiological feature in schizophrenia patients. This study aims to determine the impact of a single-nucleotide polymorphism (SNP) in the TNF-alpha gene promoter on the susceptibility, onset age, and cognitive function of schizophrenia. METHOD: The SNP -1031T>C in the TNF-alpha gene was genotyped in 905 patients and 571 healthy controls. The Positive and Negative Syndrome Scale (PANSS) was used to assess the schizophrenia symptoms and the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) for cognitive function. RESULTS: There was no significant difference in allele or genotype distribution of the SNP -1031T>C between patients and controls (p = .85, p = .98). This polymorphism had no significant genotypic effect on the symptomatology assessed by the PANSS. Interestingly, this polymorphism was significantly correlated with onset age in schizophrenia patients (p = .004). We found an earlier onset age in patients with the TT genotype compared to those with the CT and CC genotypes (both p < .05). Moreover, there were significant genotypic effects on the immediate memory index, visuospatial/constructional index, and RBANS total score (all p < 0.05) in the patient group. CONCLUSIONS: Our results suggest that the SNP -1031T>C of the TNF-alpha gene may not be associated with susceptibility to schizophrenia but possibly acts as a modulator for its onset age as well as for cognitive deficits. (PsycINFO Database Record (c) 2019 APA, all rights reserved).
Subject(s)
Genetic Predisposition to Disease , Polymorphism, Single Nucleotide , Schizophrenia/genetics , Tumor Necrosis Factor-alpha/genetics , Adult , Age of Onset , Aged , Alleles , Asian People , China , Cognition/physiology , Female , Genotype , Humans , Male , Middle Aged , Neuropsychological Tests , Promoter Regions, Genetic , Young AdultABSTRACT
Recent compelling research has demonstrated a pathophysiologic role for proinflammatory cytokines of microglial origin in decreasing neurocognitive function. Psychiatric diseases are already known to have reduced cognitive function and are also associated with increased inflammation. To elaborate on these data, our study aims to investigate how a particular polymorphism of the tumor necrosis factor gene, TNF-α -1031T/C, affects neurocognitive performance in patients with schizophrenia. We recruited 905 patients with schizophrenia and 571 healthy control subjects. We employed the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) to test for neurocognitive function and the positive and negative syndrome scale to evaluate schizophrenia severity. The -1031T/C polymorphism was genotyped in both healthy controls and schizophrenic patients. Our results demonstrate that patients with the C allele (either T/C or C/C) possessed increased immediate memory index, visuospatial/constructional index, and RBANS total scores as compared to patients without it (p < .05). In healthy controls, there was no significant difference across genotypes (p > .05). Our findings demonstrate that the TNF-α -1031T/C polymorphism may not play a role in the susceptibility of schizophrenia itself, but may be involved in the cognitive deficits of schizophrenia. This suggests an important role for cytokine signaling in mediating the severity of cognitive dysfunction in schizophrenia.
Subject(s)
Cognitive Dysfunction/genetics , Schizophrenia/genetics , Tumor Necrosis Factor-alpha/genetics , Adult , Alleles , Case-Control Studies , Cognition/physiology , Cognition Disorders/genetics , Female , Gene Frequency/genetics , Genetic Predisposition to Disease/genetics , Genotype , Humans , Male , Memory, Short-Term , Middle Aged , Neuropsychological Tests , Polymorphism, Single Nucleotide/genetics , Psychiatric Status Rating Scales , Schizophrenia/metabolism , Schizophrenic Psychology , Tumor Necrosis Factor-alpha/physiologyABSTRACT
Evidence shows that BDNF may regulate activity-dependent forms of synaptic plasticity underlying learning and memory. Previous studies reported low BDNF levels and cognitive impairment in the early stage of schizophrenia. Our current study aimed to explore the association between serum BDNF and cognitive functions in first-episode drug-naïve (FEDN) patients with schizophrenia, which has been under-investigated. We recruited 80 FEDN patients and 80 healthy controls and examined the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) and serum BDNF in both groups. Patient psychopathology was assessed using the Positive and Negative Syndrome Scale (PANSS). BDNF levels were significantly lower in patients compared to controls (pâ¯<â¯0.001). The RBANS total score and nearly all indexes (all pâ¯<â¯0.001) except for visuospatial/constructional index (pâ¯>â¯0.05) were significantly lower in patients than controls. No significant correlation was found between BDNF and any index or total scores of RBANS in either patients or healthy controls (all pâ¯>â¯0.05). However, the PANSS negative subscale score were negatively associated with both the immediate memory and language indexes (both pâ¯<â¯0.005). Our findings suggest that excessive cognitive impairments are present in the early stage of schizophrenia. Low BDNF may contribute to the pathogenesis of schizophrenia, but maybe not to its cognitive impairments.
Subject(s)
Brain-Derived Neurotrophic Factor/blood , Cognitive Dysfunction/blood , Cognitive Dysfunction/diagnosis , Schizophrenia/blood , Schizophrenia/diagnosis , Adolescent , Adult , Biomarkers/blood , Cognition/physiology , Cognitive Dysfunction/epidemiology , Female , Humans , Male , Memory, Short-Term/physiology , Middle Aged , Neuropsychological Tests , Schizophrenia/epidemiology , Young AdultABSTRACT
The forkhead-box P2 (FOXP2) gene polymorphism has been reported to be involved in the susceptibility to schizophrenia; however, few studies have investigated the association between FOXP2 gene polymorphism and clinical symptoms in schizophrenia. This study investigated whether the FOXP2 gene was associated with the development and symptoms of schizophrenia in relatively genetically homogeneous Chinese population. The FOXP2 rs10447760 polymorphism was genotyped in 1069 schizophrenia inpatients and 410 healthy controls using a case-control design. The patients' psychopathology was assessed by the Positive and Negative Syndrome Scale (PANSS). We found no significant differences in the genotype and allele distributions between the patient and control groups. Interestingly, we found significant differences in PANSS total, positive symptom, and general psychopathology scores between genotypic subgroups in patients, with the higher score in patients with CC genotype than those with CT genotype (all p < 0.05). After adjusting demographic and clinical variables, the difference still remained significant for the PANSS positive symptom score and general psychopathology (both p < 0.05). Our findings suggest that the FOXP2 rs10447760 polymorphism may not contribute to the development of schizophrenia, but may contribute to the clinical symptoms of schizophrenia among Han Chinese.