A Nested Case-Control Study to Explore the Association between Immunoglobulin G N-glycans and Ischemic Stroke / 生物医学与环境科学（英文）

OBJECTIVE@#This study prospectively investigates the association between immunoglobulin G (IgG) N-glycan traits and ischemic stroke (IS) risk.@*METHODS@#A nested case-control study was conducted in the China suboptimal health cohort study, which recruited 4,313 individuals in 2013-2014. Cases were identified as patients diagnosed with IS, and controls were 1:1 matched by age and sex with cases. IgG N-glycans in baseline plasma samples were analyzed.@*RESULTS@#A total of 99 IS cases and 99 controls were included, and 24 directly measured glycan peaks (GPs) were separated from IgG N-glycans. In directly measured GPs, GP4, GP9, GP21, GP22, GP23, and GP24 were associated with the risk of IS in men after adjusting for age, waist and hip circumference, obesity, diabetes, hypertension, and dyslipidemia. Derived glycan traits representing decreased galactosylation and sialylation were associated with IS in men (FBG2S2/(FBG2 + FBG2S1 + FBG2S2): odds ratio ( OR) = 0.92, 95% confidence interval ( CI): 0.87-0.97; G1 n: OR = 0.74, 95% CI: 0.63-0.87; G0 n: OR = 1.12, 95% CI: 1.03-1.22). However, these associations were not found among women.@*CONCLUSION@#This study validated that altered IgG N-glycan traits were associated with incident IS in men, suggesting that sex discrepancies might exist in these associations.

CAR T cells redirected against tumor-specific antigen glycoforms: can low-sugar antigens guarantee a sweet success? / 医学前沿

Immune-based therapies have experienced a pronounced breakthrough in the past decades as they acquired multiple US Food and Drug Administration (FDA) approvals for various indications. To date, six chimeric antigen receptor T cell (CAR-T) therapies have been permitted for the treatment of certain patients with relapsed/refractory hematologic malignancies. However, several clinical trials of solid tumor CAR-T therapies were prematurely terminated, or they reported life-threatening treatment-related damages to healthy tissues. The simultaneous expression of target antigens by healthy organs and tumor cells is partly responsible for such toxicities. Alongside targeting tumor-specific antigens, targeting the aberrantly glycosylated glycoforms of tumor-associated antigens can also minimize the off-tumor effects of CAR-T therapies. Tn, T, and sialyl-Tn antigens have been reported to be involved in tumor progression and metastasis, and their expression results from the dysregulation of a series of glycosyltransferases and the endoplasmic reticulum protein chaperone, Cosmc. Moreover, these glycoforms have been associated with various types of cancers, including prostate, breast, colon, gastric, and lung cancers. Here, we discuss how underglycosylated antigens emerge and then detail the latest advances in the development of CAR-T-based immunotherapies that target some of such antigens.

Association between plasma IgG N-glycan profiling and suboptimal health status among undergraduates in a college in Shandong Province / 中华健康管理学杂志

Objective:To explore the association between suboptimal health status (SHS) and plasma IgG N-glycans levels among undergraduates in a college in Shandong Province.Methods:A case-control study was conducted from September to November 2017, 100 college students who underwent physical examinations at Weifang University in Shandong, were selected as study participants based on the inclasion and exclusion criteria of the study. According to the criteria of SHS, the participants were divided into an SHS group ( n=50) and a health control group ( n=50). Plasma IgG N-glycosylaton levels were analyzed by means of ultra-high liquid chromatography (UPLC), and 24 glycan peaks were obtained. The Mann-Whitney U-test and binary logistic regression analysis were performed to investigate the association between IgG N-glycans and SHS. P<0.05 was considered statistically significant. Receiver operating characteristic (ROC) curve analyses were used to evaluate the possibility of plasma IgG N-glycans being a biomarker of SHS. Results:The results of univariate and multivariate analysis showed that GP17 was associated with SHS ( P<0.05), and the relative abundance of initial glycan peaks (GP17) was higher in the SHS group compared with the control group. ROC curve analysis showed that the area under the curve (AUC) of the baseline model was 0.826 (95% confidence interval [ CI]: 0.747-0.905, P<0.001); the AUC of the glycan-based model was 0.631 (95% CI: 0.519-0.744, P=0.002), and the AUC of the combined model was 0.848 (95% CI: 0.763-0.912, P<0.001). Compared with the baseline model, the diagnostic efficiency of the combined model revealed a trend of improvement. Conclusions:The SHS of the students in a college in Shandong Province was associated with an IgG N-glycan level of GP17, which was significantly higher than that of the control group.

Comparing different domains of analysis for the characterisation of N-glycans on monoclonal antibodies / 药物分析学报

With the size of the biopharmaceutical market exponentially increasing, there is an aligned growth in the importance of data-rich analyses, not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships. In monoclonal antibodies, many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known. The importance of their function focuses analytical research efforts on the development of robust, accurate and fast methods to support drug development and quality control. Released N-glycan analysis is considered as the gold standard for glycosylation characterisation;however, it is not the only method for quantitative analysis of glycoform heterogeneity. In this study, ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies. While observing good comparability between the quantitative results generated, it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow ac-cording to application and the desired depth of data generated.

Molecular characteristics of polysulfated fractions from the green seaweed Caulerpa cupressoides and actions on thrombin generation in vitro

Caulerpa cupressoides produces sulfated polysaccharides (Cc-SPs) with serpin-dependent anticoagulant effect, but their actions on thrombin generation (TG) are unknown. This study aimed to partially characterize Cc-SPs and examine their potential as modulators of TG. Infrared analysis characterized extract containing three ulvan fractions (Cc-SP1, -SP2 and -SP3) separated by DEAEcellulose chromatography, with differences in the relative proportions of sulfate (10.99-18.38%) and total sugars (46.59-51.12%), without presenting proteins. Charge density patterns and nonSPs varying from 8 to > 100 kDa on agarose and polyacrylamide gel electrophoresis by sequential staining with toluidine blue and stains-all were also confirmed by gel permeation chromatography. The molecular weight of Cc-SP2 was not altered after treatment with 0.4 M HCl up to 5 h. Only Cc-SP2 altered the activated partial thromboplastin time (15 ± 0.3 IU) vs. heparin (193 IU) and abolished at high concentrations (> 4.1 μg) TG by intrinsic pathway in 60-fold diluted human plasma, while at 4.1 μg attenuated TG by 33.87% delaying the lag phase (32 min.) vs. control (28 min.). Cc-SP2 induced concentration-dependent TG in system without cephalin. Heparin abolished TG at 4.15-fold lower amount, but did not stimulate TG. Therefore, Cc-SPs express dual effects on thrombosis in vitro.

Structural and physical-chemical analyses of sulfated polysaccharidesfrom the sea lettuce Ulva lactuca and their effects on thrombin generation / Análises estrutural e físico-química de polissacarídeos sulfatados da alface do mar Ulva lactuca e seus efeitos sobre geração de trombina

Ulva lactuca (Chlorophyceae) has biotechnologically-important sulfated-polysaccharides (Ul-SPs), but their potentials on thrombin generation (TG) are unknown. This study analyzed the structural and physicalchemical features of the Ul-SPs as modulators of TG. Proteolytic digestion yielded (13.13%) extract containing sulfate (20.43%) and total sugars (65.72%), besides ulvan consisting of rhamnose, xylose, glucose, glucuronic acid and α-/β-types glycosidic linkages as characterized by one-/two-dimensions nuclear magnetic resonance (NMR) experiments. Fractionation of the Ul-SPs by DEAE-cellulose chromatography yielded Ul-SP1 and Ul-SP2 (0.50 and 0.75 M NaCl, respectively) showing sulfation (15.72-18.04%) and total sugars (59.73-60.58%) consistent with the charge density pattern by combination of agarose/polyacrylamide gel eletrophoresis using sequential staining with toluidine blue and stains-all, although with slight differences in their sizes (40 and >100 kDa, respectively). By both activated partial thromboplastin time (APTT) and prothrombin time (PT) tests, anticoagulation of the fractions was virtually detected by APTT (0.39 and 0.43 IU, respectively) against heparin (193 IU). Fractions acted differently on both intrinsic/extrinsic pathways in TG using 60-fold diluted human plasma, with 50% efficacies up to 8.3 μg, whereas at high concentrations suggested intrinsic hypercoagulability since heparin abolished both systems at low amounts. Ul-SPs block TG, but predicting thrombosis in increasing doses.

Clorofícea Ulva lactuca possui polissacarídeos sulfatados (Ul-PSs) importantes biotecnologicamente, porém são desconhecidos seus potenciais sobre geração de trombina (GT). Analisaram-se as características estruturais e físico-químicas dos Ul-PSs como moduladores de GT. Digestão proteolítica rendeu (13,13%) extrato contendo sulfato (20,43%) e açúcares totais (65,72%), além de ulvana, como caracterizada por experimentos de ressonância magnética nuclear uni-/bi-dimensionais, consistindo de ramnose, xilose, glucose, ácido glucurônico e ligações glicosídicas tipos-α/-β. Fracionamento dos Ul-PSs por cromatografia de DEAE-celulose rendeu Ul-PS1 e Ul-PS2 (0,50 e 0,75 M de NaCl, respectivamente) mostrando sulfatação (15,72-18,04%) e açúcares totais (59,73- 60,58%) consistentes com o grau de densidade de carga por combinação de eletroforese em gel de agarose/poliacrilamida usando coramento sequencial com azul de toluidina e "stains-all", embora com diferenças quanto aos seus tamanhos (40 e >100 kDa, respectivamente). Por ambos os testes do tempo de tromboplastina parcial ativada (TTPA) e do tempo de protrombina, anticoagulação das frações foi detectada virtualmente pelo TTPA (0,39 e 0,43 UI, respectivamente) frente heparina (193 UI). Frações atuaram diferentemente sobre ambas as vias intrínsica/extrínsica na GT usando plasma humano diluído 60 vezes, com eficácias de 50% até 8,3 μg, enquanto em concentrações maiores sugeriram hipercoagulabilidade intrínsica visto que heparina aboliu ambos os sistemas em quantidades baixas. Ul-PSs bloqueiam GT, porém prevendo trombose em doses crescentes.

In vitro inactivation of thrombin generation by polysulfated fractions isolated from the tropical coenocytic green seaweed Caulerpa racemosa (Caulerpaceae, Bryopsidales) / Inativação in vitro da geração de trombina por frações polissulfatadas isoladas da macroalga verde cenocítica tropical Caulerpa racemosa (Caulerpaceae; Bryopsidales)

Pharmacological efficacy of Caulerpa racemosa (Chlorophyta) sulfated polysaccharidic (SPs) fractions (F I→III) on models of coagulation and inflammation has been demonstrated, but not their effects on thrombin generation (TG). This study examined fractions for composition and physical-chemical characteristics and in vitro inactivation of TG by F I and F II in 60-fold diluted human plasma using continuous method. Papain-extraction yield of 0.7% revealed F I→III by DEAE-cellulose chromatography, with differences among the relative proportions of sulfate (17.37-24.00%), total sugars (30.03-48.34%) and absence of proteins. Charge density patterns and molecular sizes > 100 kDa of the fractions were verified by both agarose/polyacrylamide analyses, respectively. These electrophoreses combined with toluidine blue/Stains-All also indicated nonSPs. Anticoagulant effects of 4.76 (F I), 12.00 (F II) and 2.32 (F II) IU mg-1 by activated partial thromboplastin time test were recorded against heparin (193 IU mg-1), without changes in prothrombin time. Diluted plasma treated with F I and F II reduced concentration-dependent and sulfation pattern TG by both intrinsic and extrinsic pathways, with 50% inactivation by intrinsic pathway of F II even at 4.1 µg. Heparin abolished TG at least 4-fold lower. Therefore, C. racemosa produces SPs with TG inhibition.

Eficácia farmacológica de frações (F I→III) polissacarídicas sulfatadas (PSs) da Chlorophyta Caulerpa racemosa sobre modelos de coagulação e inflamação tem sido demonstrada, exceto seus efeitos sobre geração de trombina (GT). Examinaram-se frações quanto à composição, características físico-químicas e inativação in vitro de GT por F I e F II, em plasma humano diluído 60 vezes usando método contínuo. Rendimento de extração-papaína (0,7%) revelou, por cromatografia de DEAE -celulose, F I→III com diferenças entre as proporções relativas de sulfato (17,37-24,00%), açúcares totais (30,03-48,34%) e ausência de proteínas. Foram verificados, por ambas as análises agarose/poliacrilamida, graus de densidade de carga e tamanhos moleculares > 100 kDa das frações, respectivamente. Também essas eletroforeses, combinadas com azul de toluidina/Stains-All, indicaram polissacarídeos não sulfatados. Foram registrados, pelo teste do tempo de tromboplastina parcial ativada, efeitos anticoagulantes de 4,76 (F I), 12,00 (F II) e 2,32 (F II) UI mg-1 contra heparina (193 UI mg- 1), porém não modificando tempo de protrombina. Plasma diluído tratado com F I e F II reduziu GT por ambas as vias intrinsíca/extrínsica, dependente de concentração e grau de sulfatação, com F II em 4,1 µg apresentando eficácia de 50% pela via intrínsica. Heparina, quatro vezes menos, aboliu GT. Portanto, C. racemosa produz PSs com inibição de GT.

HEPARAN SULFATE AND CHONDROITIN SULFATE: GENERAL ASPECTS OF THEIR PARTICIPATION DURING Plasmodium falciparum DEVELOPMENT / SULFATO DE HEPARÁN Y SULFATO DE CONDROITINA: ASPECTOS GENERALES DE SU PARTICIPACIÓN DURANTE EL DESARROLLO DE Plasmodium falciparum

Background: Malaria is one of the most important infectious diseases worldwide due to its high morbidity and mortality rates every year in tropical countries. Despite efforts in malaria research, several mechanisms underlying hostparasite interactions remain unclear, which is a big obstacle for the management and control of malaria. Recently, numerous studies have attempted to provide a better understanding of the physiopathological mechanisms to assist in the design of new drugs, vaccines and transmission blocking agents. These research topics have indicated that glycans are key molecules in the life cycle of the malarial parasites. The aim of this review is to highlight the relevance of glycans for the development and transmission of Plasmodium and to use that information as a valuable research tool to fight malaria. Because glycans play roles in parasite invasion and interactions with the mosquito host, both of which are part of "parasite development", this review seeks to specify the role of glycans in parasite development. Methods: This review was mainly based on research articles published between 1985 and 2015 that were obtained from the PubMed and Embase databases. The keywords used in this search were sulfated glycans, malaria, Anopheles and Plasmodium. Conclusions: Sulfated glycoconjugates are intimately linked to the development, transmission and survival of Plasmodium in the intermediate and definitive hosts. A better understanding of the role of sulfated glycoconjugates in malaria infection would permit the development of new therapeutic strategies and the design of strategies to inhibit parasite transmission.

La malaria es considerada una de las enfermedades infecciosas de mayor importancia alrededor del mundo debido a la alta morbimortalidad que causa cada año en países tropicales. A pesar de los esfuerzos de investigación en malaria, muchos de los mecanismos que entrañan las interacciones hospedero-parásito aún no son claros, lo que constituye un gran obstáculo en el manejo y control de la malaria. Numerosos estudios se han llevado a cabo en los últimos años en busca de una mejor comprensión de los mecanismos fisiopatológicos, diseño de nuevas drogas, diseño de una vacuna y bloqueo de la transmisión. En todos estos temas de investigación, un elemento común son los glicanos como moléculas clave en el ciclo de vida de los parásitos de la malaria. El objetivo de esta revisión es mostrar como los glicanos se necesitan para el desarrollo y la transmisión de Plasmodium y como esta información resulta ser una valiosa herramienta en la investigación para combatir la malaria. Métodos: La presente revisión se basó principalmente en artículos originales publicados entre 1985 y 2015, obtenidos de las bases de datos PubMed y EmBase. La búsqueda fue hecha en inglés y se usaron las palabras clave: glicanos sulfatados, malaria, Anopheles y Plasmodium. Conclusión: Los glicoconjugados sulfatados están íntimamente vinculados al desarrollo, la transmisión y la supervivencia de Plasmodium, tanto en el hospedero intermediario como en el hospedero definitivo. Una mejor comprensión del rol de los glicoconjugados sulfatados en la infección malárica permitiría el desarrollo de nuevas alternativas terapéuticas, así como el diseño de estrategias para inhibir la transmisión.

An anti-dengue and anti-herpetic polysulfated fraction isolated from the coenocytic green seaweed Caulerpa cupressoides inhibits thrombin generation in vitro / Uma fração polissulfatada anti- dengue e anti- herpética isolada da alga marinha verde cenocítica Caulerpa cupressoides inibe geração de trombina in vitro

The antiviral potency of Caulerpa cupressoides (Chlorophyta) sulfated polysaccharidic fractions (Cc -SP1→3) associated with thrombin generation (TG) assay is unknown. This study analyzed the structure of Cc-SP1 and its effects against herpes viruses (HSV-1 and HSV-2), dengue-virus (DENV-1) , human-metapneumovirus (HMPV) and adenovirus (AdV) in Vero and mosquito C6/36 cell-lines and as inhibitor of TG in 60- fold diluted human plasma using continuous system. Infrared analysis indicated ulvan containing 11% sulfate, 40.16% total sugars and 6% uronic acid. Procedures of agarose/polyacrylamide gels electrophoresis revealed two major components presenting sulfate and non SPs, with molecular dispersion ranging from 8 to > 100 kDa, respectively, as confirmed by gel permeation chromatography. Cc-SP1 did not cause cytotoxicity up to 1000 µg mL-1 and was more effective inhibitor of DENV-1 (96%, 0.35 µg mL-1) compared with HSV-1 (90%, 27 µg mL- 1) and HSV-2 (99.9%, 0.9 µg mL-1) in Vero cell-line, whose selectivity indexes were > 714, > 9.2 and > 131, respectively, but was inactive against HMPV, AdV and C6/36 cell-line using plate reduction assay. Cc-SP1 required concentration 20.8-fold higher of SPs than heparin for abolishes intrinsically TG, but at concentrations so far the anti-HSV-1. Therefore, TG assay provides data to guide studies of Cc-SP1 on anticoagulant/antiviral effects.

A potência antiviral de frações polissacarídicas sulfatadas de Caulerpa cupressoides (Chlorophyta) (Cc-PS1→3) é desconhecida e associada com ensaio de geração de trombina (GT). Analisaram-se de Cc-PS1 estrutura e efeitos contra herpesviroses (HSV-1 e HSV-2), vírus da dengue (DENV-1), metapneumovírus-humano (MPVH) e adenovírus (AdV) em linhagens de células Vero e de mosquito C6/36 e como inibidora de GT em plasma humano diluído 60 vezes usando sistema contínuo. Análise de infravermelho indicou 'ulvana' contendo sulfato (11%), açúcares totais (40,16%) e ácido urônico (6%). Procedimentos de electroforese em géis de agarose/poliacrilamida revelaram dois componentes majoritários apresentando sulfato e polissacarídeos não sulfatados, com dispersão molecular variando de 8 a > 100kDa, respectivamente, confirmada por cromatografia de permeação em gel. Cc-PS1 não causou citotoxicidade até 1000 µg mL-1 e se mostrou, em linhagem de célula Vero usando ensaio de redução de placas, inibidora efetiva de DENV-1 (96%; 0,35 µg mL-1) comparada com HSV-1 (90%; 27 µg mL-1) e HSV-2 (99,9%; 0,9 µg mL-1), cujos índices de seletividade foram > 714; > 9,2 e > 278, respectivamente, enquanto, demonstrou-se ineficaz sobre HMPV, AdV e linhagem de célula C6/36. Foi requerida concentração 20,8 vezes maior de Cc-PS1 que heparina para abolir GT intrinsicamente, mas em concentrações diferentes da anti-HSV-1. Portanto, ensaio de GT fornece dados para estudos direcionados sobre efeitos anticoagulante/antiviral de Cc-PS1.

Comparison between CA19-9,CEA and specific N-Glycans in early middle stage pancreatic carcinoma serum / 实用肿瘤学杂志

Objective To test and compare CA19-9,CEA with Specific N-Glycans in early-middle stage pancreatic carcinoma serum to find a better tumor marker in early pancreatic carcinoma .Methods To find the difference of N-Glycans structure ,DSA-FACE was performed in 35 early middle stage pancreatic carcinoma patients and 50 healthy human .Meanwhile ,we tested CA19-9 and CEA in the pancreatic carcinoma patients at the same stage.Result The serum N -glycan profiles of pancreatic carcinoma was identified by the DSA -FACE technique.The results showed that between pancreatic carcinoma patients and healthy persons ,there were significant differences in N -glycans.The peak 13,14 and 17 were the most significant peaks which would be most likely picked as a new tumor marker of pancreatic carcinoma .Taking log(p14 ×p17/p13)as indicator of the ROC curve analysis,and the area under ROC curve was 0.799 ±0.050 with 84.9% sensitivity and 68% speci-ficity.Meanwhile,the sensitivity of CA19-9 was 61.2%,the sensitivity of CEA was 11.7%.Conclusion The Peak 13,14 and 17 are the most significant peaks which would be picked as a new tumor marker of pancreatic carcinoma.Espicially,its sensitivity is superior to CA19-9、CEA for early middle stage patients .

Heparin Co-factor II Thrombin Complex as a Biomarker for Mucopolysaccharidosis: Indian Experience.

Background: Serum heparin cofactor II-thrombin complex (HCII-T) is an emerging biomarker for mucopolysaccharidosis disease (MPS I and MPS II). Methods: Seventeen cases (6 MPS I and 11 MPS II) and sixty healthy controls were enrolled in study, conducted from September 2008 to December 2012. The mean ± SD age of MPS1 (n=6, 5 males) and MPS II was 7.02 ± 3.25 and 5.2 ± 2.15 years, respectively. Disease status was confirmed by clinical features and enzyme assay. Urinary glycosaminoglycans were measured in spot urine samples and expressed in relation to creatinine content. HCIIT measurement was done using sandwich ELISA at enrolment and after 12 and 24 months of recruitment. Results: Urinary glycosaminoglycans and HCIIT were elevated in all patients compared to their healthy controls. Both markers could not discriminate between the type of mucopolysaccharidosis. Conclusion: Heparin Cofactor II Thrombin Complex is a good biomarker for mucopolysaccharidosis I and II.

Significance of postoperative structural changes in serum N-glycans in pancreatic cancer patients / 临床肝胆病杂志

Objective To investigate the structural changes in specific serum N-glycans in pancreatic cancer patients and to identify the specific serum maker of pancreatic cancer.Methods The pancreatic cancer patients who visited the Third Affiliated Hospital of Harbin Medical University from June 201 1 to December 2013 were assigned to preoperative serum group (123 cases)and postoperative serum group (78 cases);healthy controls whose serum samples were collected in the Physical Examination Center were selected as control serum group (271 cases).DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis (DSA-FACE)was used to analyze serum N -glycans and compare them between the three groups.Results The serum N-glycan profiles in pancreatic cancer patients were identified by DSA-FACE.The results indicated that N-glycan peak 8 in preoperative serum group was significantly lower than those in control serum group (t=2.735,P<0.05)and postoperative serum group (P<0.05),but no significant difference was found between the postoperative serum group and control serum group.Conclusion N-glycan peak 8 can be considered as a serum marker of pancreatic cancer.

Lectinhistoquímica del granuloma inducido por el bacilo de Calmette Guérin en Piaractus mesopotamicus / Lectinhistochemical staining of granuloma induced by bacillus Calmette-Guerin in Piaractus mesopotamicus

Objetive. This study was conducted to evaluate, by means of lectinhistochemistry (LHC), the expression of carbohydrates in granulomas induced by the bacillus Calmette-Guerin (BCG) in muscle tissue of Piaractus mesopotamicus after 33 days. Material and methods. Histological sections with 3 μm thick were incubated with the following lectins :WGA (Wheat germ agglutinin), DBA (Dolichos biflorus agglutinin) and HPA (Helix pomatia agglutinin), and the results were evaluated by light microscopy. Results. Acid fast bacilli were stained by Ziehl Neelsen (ZN) and strong labeled by WGA in the cytoplasm of macrophages. Labeling with DBA was intense in fibroblasts and weak in macrophages. On the other hand, HPA binding was stronger in macrophages, especially in those that were in close contact with epithelioid cells, without evidence of binding to fibroblasts. The epithelioid cells were not labeled by the used lectins, but they were identified by Hematoxilin-Eosin (HE). The lectins labeled specific type saccharides in glycoproteins, as N-acetylglucosamine present in bacilli and macrophages, as well as N-acetyl-galactosamine in macrophages. The control group showed no inflammation or lectin binding. Conclusions. This technique may be useful in identifying receptors for WGA, DBA and the HPA lectins in epithelioid granuloma induced by BCG in P. mesopotamicus.

El presente estudio fue realizado para evaluar por medio de lectinhistoquímica (LHC), la expresión de carbohidratos en granulomas inducidos por el bacilo de Calmette-Guérin (BCG) en músculo de Piaractus mesopotamicus después de 33 días. Materiales y métodos. Cortes histológicos de 3 µm de grosor fueron incubados con las siguientes lectinas: WGA (Wheat germ aglutinin), DBA (Dolichos biflorus agglutin) y HPA (Helix pomatia agglutinin), y los resultados evaluados por medio de microscopia de luz. Resultados. Bacilos ácido resistentes fueron identificados por la tinción de Ziehl Neelsen(ZN). Se observó un marcaje intenso con WGA en el citoplasma de macrófagos. El marcaje con DBA fue intenso en fibroblastos y débil en macrófagos. Con la lectina HPA el marcaje fue intenso en macrófagos, principalmente en los que estaban en estrecho contacto con las células epitelióides, externamente se observó marcaje débil en fibroblastos. Las células epitelióides no fueron marcadas por las lectinas, pero fueron identificadas con la tinción de Hematoxilina-Eosina (HE). Las lectinas tuvieron un tipo de marcaje específico en algunos monosacáridos, como N-acetilglucosamina presente en los bacilos y en macrófagos, y N-acetilgalactosamina en macrófagos. En el grupo control no fue observada inflamación así como tampoco marcaje con las lectinas. Conclusiones. Esta técnica resultó eficiente en la identificación de receptores para las lectinas WGA, DBA y HPA en el granuloma epitelióide inducido por BCG en P. mesopotamicus.

Structure Characteristics of N-linked Glycans in Salivary Glycoproteins of Chronic Gastritis Patients with Spleen Deficiency / 广州中医药大学学报

【Objective】To analyze the structure of N-linked glycans in salivary glycoproteins and amylase of chronic gastritis(CG)patients with spleen deficiency after stimulation of acid,and to explore the relationship of N-linked glycans structure with salivary amylase activity and splenic hypofunction.【Methods】Iodine-spectrophotometry was used to detect the amylase activity in 47 CG patients with spleen deficiency,15 CG patients with dampness-heat in the spleen and stomach,and 12 healthy volunteers before and after acid stimulation.Method of concanacalin A(ConA) affinity chromatography was used to separate total salivary glycoproteins and amylase and to analyze N-linked glycans structure in 27 CG patients with spleen deficiency,14 CG patients with dampnessheat in the spleen and stomach,and 5 healthy volunteers after acid stimulation.ConA affinity reversed-phase two-dimension chromatography was applied to separate total salivary glycoproteins and to analyze the constitute and distribution of total salivary glycoproteins with different N-linked glycans in 13 CG patients with spleen deficiency,6 CG patients with dampness-heat in the spleen and stomach,and 5 healthy volunteers after acid stimulation.【Results】After acid stimulation,salivary amylase activity in CG with spleen deficiency decreased as compared with that before acid stimulation(P

Progress in congenital disorders of glycosylation / 中国病理生理杂志

Congenital disorders of glycosylation(CDG) are a rapidly growing group of genetic diseases that are due to defects in the synthesis of glycans and in the attachment of glycans to other compounds. Most CDG are multisystemic diseases often involving severe psychomotor retardation. The CDG causing sialic acid deficiency of N-glycans can be diagnosed by isoelectric focusing of serum transferrin. Glycan structural analysis, yeast genetics and knockout animal models are essential tools in the elucidation of novel CDG. In this review, we focus on the current knowledge of the pathogenesis and identification of nine primary glycosylation diseases.