Vanillic Acid Inhibits Lung Carcinogenesis By Modulates Glycoprotein Abnormalities, Membrane-Bound Enzymes, And Inflammatory Markers

Objective: The current plan was accompanied to explicate the possible protective role of vanillic acid (VA), on modification in lipid peroxidation, inflammatory cytokines, membrane-bound enzymes, and glycoconjugates during B(a)P induced lung cancer in Swiss albino mice. Methods: Benzo(a)pyrene was administered orally (50 mg/kg b. wt) to induce lung cancer in Swiss albino mice. lipid peroxidation, serum marker enzymes, inflammatory cytokines, membrane-bound ATPases and protein-bound carbohydrate components (Hexose, hexosamine, sialic acid and fucose) and Mast cells and PAS staining were carried out. Results: Lung cancer possessing animals exhibited increased levels of lipid peroxidation, ADA, AHH, γ-GT, 5’-NT, LDH, cytokines such as TNF-α and IL-1β, protein-bound carbohydrate components (protein-hexose, hexosamine, sialic acid, and fucose) also diminished activity of membrane-bound ATPases (Na+/K+ATPases, Ca2+ATPases, and Mg2+ATPase). Treatment with VA significantly ameliorated all these activities. Conclusion: Overall, the present study evidence to the VA has effective anti-inflammatory in addition to free radical scavenging activity for the duration of lung carcinogenesis in Swiss albino mice.

Leishmania infantum induces high phagocytic capacity and intracellular nitric oxide production by human proinflammatory monocyte

BACKGROUND The mechanism of resistance to SbIII in Leishmania is complex, multifactorial and involves not only biochemical mechanisms, but also other elements, such as the immune system of the host. OBJECTIVES In this study, putative changes in the immunological profile of human monocytes infected with wild-type (WT) and antimony (SbIII)-resistant Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum lines were evaluated. METHODS Susceptibility assays WT and SbIII-resistant L. braziliensis and L. infantum were performed using lines THP-1 human monocytic lineage. Phagocytic capacity, cytokine profile, intracellular nitric oxide (NO) production and surface carbohydrate residues profile were performed in peripheral blood monocytes by flow cytometry. FINDINGS The phagocytic capacity and intracellular NO production by classical (CD14++CD16-) and proinflammatory (CD14++CD16+) monocytes were higher in the presence of L. infantum lines compared to L. braziliensis lines. The results also highlight proinflammatory monocytes as the cellular subpopulation of major relevance in a phagocytosis event and NO expression. It is important to note that L. infantum induced a proinflammatory cytokine profile characterised by higher levels of TNF-α in culture supernatant than L. braziliensis. Conversely, both Leishmania lines induce high levels of IL-6 in culture supernatant. Analysis of the expression profile of surface carbohydrates showed that L. braziliensis presents 4.3-fold higher expression of galactose(β1,4)N-acetylglucosamine than L. infantum line. Interestingly, the expression level of α-N-acetylgalactosamine residues was 2-fold lower in the SbIII-resistant L. braziliensis line than its counterpart WT line, indicating differences in surface glycoconjugates between these lines. MAIN CONCLUSIONS Our results showed that L. braziliensis and L. infantum induce different innate immune responses and a highly inflammatory profile, which is characteristic of infection by L. infantum, the species associated with visceral disease.

Scedosporium apiospermum, Scedosporium aurantiacum, Scedosporium minutisporum and Lomentospora prolificans: a comparative study of surface molecules produced by conidial and germinated conidial cells

BACKGROUND Scedosporium/Lomentospora species are opportunistic mould pathogens, presenting notable antifungal resistance. OBJECTIVES/METHODS We analysed the conidia and germinated conidia of S. apiospermum (Sap), S. aurantiacum (Sau), S. minutisporum (Smi) and L. prolificans (Lpr) by scanning electron microscopy and exposition of surface molecules by fluorescence microscopy. FINDINGS Conidia of Sap, Smi and Sau had oval, ellipsoidal and cylindrical shape, respectively, with several irregularities surrounding all surface areas, whereas Lpr conidia were rounded with a smooth surface. The germination of Sap occurred at the conidial bottom, while Smi and Sau germination primarily occurred at the centre of the conidial cell, and Lpr germination initiated at any part of the conidial surface. The staining of N-acetylglucosamine-containing molecules by fluorescein-labelled WGA primarily occurred during the germination of all studied fungi and in the conidial scars, which is the primary location of germination. Calcofluor white, which recognises the polysaccharide chitin, strongly stained the conidial cells and, to a lesser extent, the germination. Both mannose-rich glycoconjugates (evidenced by fluoresceinated-ConA) and cell wall externally located polypeptides presented distinct surface locations and expression according to both morphotypes and fungal species. In contrast, sialic acid and galactose-containing structures were not detected at fungal surfaces. MAIN CONCLUSIONS The present study demonstrated the differential production/exposition of surface molecules on distinct morphotypes of Scedosporium/Lomentospora species.

Histochemical differences along the intestine of Corydoras paleatus (Siluriformes: Callichthyidae) / Diferencias histoquímicas a lo largo del intestino de Corydoras paleatus (Siluriformes: Callichthyidae)

AbstractThe Neotropical catfish Corydoras paleatus is a facultative air-breather and the caudal half of the intestine is involved in gas exchange. In South America, air-breathing fishes are found in tropical or sub-tropical freshwaters where the probability of hypoxia is high. The aim of this study was to characterize by traditional histochemical and lectinhistochemical methods the pattern of carbohydrate in the intestinal mucosa. Intestine samples were taken from 25 healthy adult specimens collected in Buenos Aires (Argentina). Samples were fixed by immersion in 10 % buffered formalin and routinely processed and embedded in paraffin wax. Subsequently, these sections were incubated in the biotinylated lectins battery. Labeled Streptavidin-Biotin (LSAB) system was used for detection, diaminobenzidine as chromogen and haematoxylin as a contrast. To locate and distinguish glycoconjugates (GCs) of the globet cells, we used the following histochemical methods: PAS; PAS*S; KOH/ PA*S; PA/Bh/KOH/PAS; KOH/PA*/Bh/PAS; Alcian Blue and Toluidine Blue at different pHs. Microscopically, the general structure of vertebrate intestine was observed and showed all the cell types characteristic of the intestinal epithelium. The cranial sector of catfish intestine is a site of digestion and absorption and its structure is similar to other fish groups. In contrast, enterocytes of the caudal portion are low cuboidal cells; and between these, globet cells and capillaries are observed, these latter may reach the mucosal lumen. Underlying the epithelium, observed a well-developed lamina propria-submucosa made of connective tissue; this layer was highly vascularized and did not exhibit glands. According to histochemistry, the diverse GCs elaborated and secreted in the intestine are associated with specific functions in relation to their physiological significance, with special reference to their role in lubrication, buffering effect and prevention of proteolytic damage to the epithelium together with other biological processes, such as osmoregulation and ion exchange. The lectinhistochemical analysis of the intestinal mucosa reveals the presence of terminal residues of glucose, mannose and galactose. In conclusion, this study has shown that GCs synthesized in the intestine of C. paleatus exhibit a high level of histochemical complexity and that the lectin binding pattern of the intestinal mucosa is characteristic of each species and the variations are related with the multiple functions performed by the mucus in the digestive tract. The information generated here may be a relevant biological tool for comparing and analyzing the possible glycosidic changes in the intestinal mucus under different conditions, such as changes in diet or different pathological stages. Rev. Biol. Trop. 64 (1): 327-340. Epub 2016 March 01.

ResumenEl pez neotropical Corydoras paleatus, de respiración aérea de tipo facultativa, utiliza el sector caudal del intestino para el intercambio gaseoso. En América del Sur, los peces con respiración aérea se encuentran en las aguas dulceacuícolas tropicales y subtropicales, donde la probabilidad de hipoxia es alta. El objetivo de este trabajo fue caracterizar mediante técnicas histoquímicas tradicionales y de lectinhistoquímica el patrón de carbohidratos de la mucosa intestinal. Para ello se utilizaron muestras de intestino de 25 ejemplares sanos adultos recolectados en la provincia de Buenos Aires (Argentina). Las muestras fueron fijadas en formol amortiguado al 10 % y se procesaron para su inclusión en parafina. Posteriormente, los cortes fueron incubados con una batería de lectinas biotiniladas. Se utilizó el sistema de marcado con estreptavidina-biotina (LSAB) para su detección, diaminobencidina como cromógeno y hematoxilina como colorante de contraste. Para localizar y diferenciar los glicoconjugados (GCs) de las células caliciformes, se utilizaron las siguientes técnicas histoquímicas: PAS, PAS*S, PAPS, KOH/PA*S, PA/Bh/KOH/PAS, KOH/PA*/Bh/PAS, Azul Alcian y Azul de Toluidina a diferentes pHs. Microscópicamente, se observa la estructura general del intestino de los vertebrados y el epitelio intestinal presenta todos los tipos celulares característicos de esta región. El sector craneal del intestino de este teleósteo, es el sitio de digestión y absorción, y posee una estructura similar a la de otros grupos de peces. En cambio, los enterocitos de la porción caudal, son células cúbicas bajas, entre ellos se observan células caliciformes y capilares sanguíneas que llegan hasta el lumen de la mucosa. Por fuera del epitelio, se observa una lámina propia-submucosa muy desarrollada compuesta por tejido conectivo, altamente vascularizada que no presenta glándulas. De acuerdo con las técnicas histoquímicas, los diversos GCs elaborados y secretados por la mucosa intestinal se encuentran asociados con funciones específicas de importancia fisiológica, como su rol en la lubricación, su efecto amortiguador y la prevención de daños proteolíticos del epitelio junto con otros procesos biológicos, tales como la osmorregulación y el intercambio iónico. El análisis lectinhistoquímico de la mucosa intestinal revela la presencia de residuos terminales de glucosa, manosa y galactosa. En conclusión, en este estudio se demuestra que los GCs sintetizados en el intestino de C. paleatus muestran un alto nivel de complejidad histoquímica y que el patrón de unión de lectina de la mucosa intestinal es característico para cada especie y las variaciones se hallan relacionadas con las múltiples funciones realizadas por el mucus en el tracto digestivo. La información brindada en este trabajo es una herramienta de relevancia biológica para comparar y analizar los posibles cambios glicosídicos del mucus intestinal bajo diferentes condiciones como los cambios en la dieta o diferentes estados patológicos.

Effect of Codonopsis and Glycyrrhiza glycoconjugates on migration and membrane potential of IEC-6 cells / 中国药理学与毒理学杂志

OBJECTIVE To observe the effect of Codonopsis and Glycyrrhiza glycoconjugates on migration and membrane potential of small intestinal epithelial cells(IEC-6),and to explore the promoting effects of Yiqi jianpi herb Codonopsis and Glycyrrhiza on gastrointestinal mucosal injury repair and the underlying mechanisms. METHODS Under normal conditions or loaded with the inhibitor of potassium channel 4-aminopyridine(4-AP),IEC-6 cells were treated with Codonopsis and Glycyrrhiza glycoconjugates (25-200 mg · L-1) for 24 h,respectively. IEC-6 cell migration was observed by the phase contrast microscope and cell membrane potential was detected by flow cytometry. RESULTS Codonopsis and Glycyrrhiza glycoconjugates (50 and 100 mg · L-1) increased the number of migrated IEC-6 cell compared with normal control group(P<0.01,P<0.05). Compared with normal control group,4-AP reduced the number of migrated IEC-6 cell(P<0.01). Codonopsis and Glycyrrhiza glycoconjugates (50-200 mg · L-1)reversed cell migration inhibited by 4-AP significantly when compared with 4-AP model group(P<0.01). The results of flow cyometry analysis showed that the cell membrane poten?tial was increased after treatment with Codonopsis and Glycyrrhiza glycoconjugates(50 mg · L-1)compared with normal control group and resulted in an increase in cell membrane hyperpolarization(P<0.01). Compared with normal control group,4-AP decreased the cell membrane potential(P<0.01)and resulted in cell membrane depolarization. Also,compared with 4-AP model group,cell membrane depolarization induced by 4-AP was reversed by treatment with Codonopsis and Glycyrrhiza glycoconjugates(100 and 200 mg·L-1). CONCLUSION Codonopsis and Glycyrrhiza glycoconjugates may promote gastrointestinal mucosal injury repair and the mechanisms may involve the activation of signaling pathways by affecting polyamine-dependent intestinal epithelial cell migration voltage-gated K+channels.

Snake venom galactoside-binding lectins: a structural and functional overview

Snake venom galactoside-binding lectins (SVgalLs) comprise a class of toxins capable of recognizing and interacting with terminal galactoside residues of glycans. In the past 35 years, since the first report on the purification of thrombolectin from Bothrops atrox snake venom, several SVgalLs from Viperidae and Elapidae snake families have been described, as has progressive improvement in the investigation of structural/functional aspects of these lectins. Moreover, the advances of techniques applied in protein-carbohydrate recognition have provided important approaches in order to screen for possible biological targets. The present review describes the efforts over the past 35 years to elucidate SVgalLs, highlighting their structure and carbohydrate recognition function involved in envenomation pathophysiology and potential biomedical applications.(AU)

Snake venom galactoside-binding lectins: a structural and functional overview

Snake venom galactoside-binding lectins (SVgalLs) comprise a class of toxins capable of recognizing and interacting with terminal galactoside residues of glycans. In the past 35 years, since the first report on the purification of thrombolectin from Bothrops atrox snake venom, several SVgalLs from Viperidae and Elapidae snake families have been described, as has progressive improvement in the investigation of structural/functional aspects of these lectins. Moreover, the advances of techniques applied in protein-carbohydrate recognition have provided important approaches in order to screen for possible biological targets. The present review describes the efforts over the past 35 years to elucidate SVgalLs, highlighting their structure and carbohydrate recognition function involved in envenomation pathophysiology and potential biomedical applications.

Effects of the 2, 4-D herbicide on gills epithelia and liver of the fish Poecilia vivipara / Efeitos do herbicida 2, 4-D no epitélio das brânquias e no fígado do peixe Poecilia vivípara

The 2,4-dichlorophenoxyacetic acid, usually named 2,4-D is one of the most widely used herbicides in the world. Acute toxicity of 2,4-D herbicide was investigated through its effects on guppies (Poecilia vivipara Bloch et Schneider 1801). Fish were exposed to the herbicide at concentrations of 10, 20 and 40µl per liter of water for 24 hours to determine its effects on gills and liver epithelia. The estimated LC50 was 34.64µl of 2,4-D per liter of water. Histochemical analyses and Feulgen's reaction were conducted to detect glycoconjugates and DNA, respectively, in gills and liver epithelia. Histochemistry revealed qualitative variations of glycoconjugates present on mucous cells and granules. The four types of mucous cells contained neutral granules, acids, or both. Increasing amounts of syalomucins were observed from the control group to the group exposed to the highest concentration of 2,4-D, suggesting increased mucous viscosity and the formation of plaques that could inhibit gas exchange and osmoregulation. Lamellar fusion observed in the group exposed to 40µl of 2,4-D suggests a defense mechanism. Hepatocytes showed vacuolization in the 10 and 20µl/L groups. The 40 µl/L group showed normal hepatocytes as well as changed ones, many Ito cells, micronuclei, and nuclear swelling. These effects may be associated with toxicity or adaptative processes to cellular stress. The data from this study indicates the importance of assessing similar risks to aquatic species and suggests that Poecilia vivipara is an adequate biological model for analysis of environmental contamination.

A toxicidade aguda do herbicida 2,4-D foi investigada através dos efeitos no peixe Poecilia vivípara (Bloch et Schneider, 1801). Grupos de peixes foram expostos ao herbicida nas concentrações de 10, 20 e 40µl por litro de água, durante 24 horas. As brânquias e o fígado foram estudados. A concentração letal média (CL50) do herbicida para a espécie em questão foi de 34,64µl/l. Foram realizadas colorações histoquímicas e coloração de Feulgen para identificar glicoconjugados e DNA, respectivamente, nos tecidos acima citados. Os métodos histoquímicos revelaram os tipos de glicoconjugados presentes nas células mucosas e nos grânulos. Os quatro tipos de células mucosas apresentaram glicoconjugados neutros, ácidos, ou ambos em um mesmo tipo celular. Observou-se a presença crescente de sialomucinas do grupo controle até o grupo exposto a maior concentração de 2,4-D, sugerindo aumento da viscosidade do muco e, consequentemente, formação de placas que impedem as trocas gasosas e a osmorregulação. A fusão lamelar observada no grupo exposto a 40µl de 2,4-D sugere ser um mecanismo de defesa. Os hepatócitos apresentaram processo de vacuolização nos grupos 10 e 20µl/l. No grupo de 40µl/l, observou-se a presença de células de Ito, micronúcleos e hepatócitos normais e outros com edema nuclear. Este estudo indica a importância da avaliação de riscos semelhantes a espécies aquáticas e sugere a espécie Poecilia vivipara como modelo biológico adequado para análises de contaminação ambiental.

Lectin histochemical aspects of the ovarian lamellae epithelium of Genypterus blacodes (Schneider, 1801) / Padrão de reação lectino-histoquímico do epitélio laminar ovariano em Genypterus blacodes (Schneider, 1801)

The composition and distribution of the glycoconjugates (GCs) secreted by the epithelium of ovarian lamellae with reference to the reproductive biology of Genypterus blacodes (Schneider, 1801) through lectin hi stochemistry is here discussed. In this species, the epithelial cells that line the ovarian cavity presented sharp morphological variations along the reproductive cycle related to the mucus secretion that accompanies oocyte ma turation. During sp awning season, residues of mannose and N-acetylglucosamine were detected in the glycocalyx of those cells using lectinhistochemistry. N- acetylgalactosamine and fucose were also observed in the same zone. The greatest variations in the lectinhistochemical pattern were found in the apical cytoplasm composition in comparison to the basal zone of the cells. The results of the present study were discussed by comparing their possible functional implications.

A composição e distribuição dos glicoconjugados (GCs) secretado pelo epitélio do ovário de lamelas com referência à biologia reprodutiva de Genypterus blacodes (Schneider, 1801) através da histoquímica com lectinas é aqui discutida. Nesta espécie, as células epiteliais que revestem a cavidade do ovário apresentou acentuada variação morfológica ao longo do ciclo reprodutivo relacionados com a secreção de muco que acompanha a maturação do oócito. Durante a época de desova, de resíduos de manose e N-acetilglicosamina foram detectados no glicocálix dessas células usando histoquímica de lectinas. N-acetilgalactosamina e fucose também foram observados na mesma zona. As maiores variações no padrão de lectinas foram encontradas na composição do citoplasma apical, em comparação com a zona basal das células. Os resultados do presente estudo foram discutidos, comparando as suas possíveis implicações funcionais.

The Properties of Glycoconjugates in the Rat Nasal Mucosa following Inhalation of Formaldehyde / 대한해부학회지

The present study was carried out to investigate the glycoconjugate properties of the nasal mucosa in the rat after inhalation of formaldehyde. Sprague-Dawley male rats were inhalated 30 ppm formaldehyde for 3 times with 3 hours exposure. The olfactory and respiratory mucosa in the nasal mucosa were taken from the animals on 3, 6,9 days and 2, 3, 4, 5 weeks after inhalation of formaldehyde. The properties of glycoconjugate of the olfactory and respiratory mucosa were investigated using nine biotinylated lectins (PSA, UEA I, PHA-L, BSL I, PNA, MAL I, DBA, BSL II or sWGA). In experimental groups, the degenerative changes of the olfactory epithelium were observed until 3 weeks after inhalation of formaldehyde, but the respiratory epithelium was no change. In control group, the olfactory cells in the olfactory epithelium reacted with PSA, UEA I, PNA, DBA, BSL II, sWGA, and the supporting cells reacted with PSA, PHA-L, PNA, MAL I, DBA, BSL II, sWGA, and Bowman's glands reacted with all the lectins. In experimental groups, the olfactory cells reacted with UEA I, DBA, and the supporting cells reacted with PHA-L, MAL I, DBA, UEA I, and the positive reaction of Bowman's glands was increased. In control group, the goblet cells in the respiratory epithelium reacted with UEA I, MAL I, and the ciliated columnar cells reacted with PSA, UEA I, PHA-L, BSL I, DBA, BSL II, sWGA, and the septal nasal glands reacted with all the lectins except UEA I. In experimental groups, the goblet cells reacted with UEA I, MAL I and PNA. Conclusively, the olfactory mucosa was shown a lot of changes in the properties of glycoconjugates following inhalation of formaldehyde, but respiratory mucosa was shown feeble change. These results suggest that there were different sugar residues of glycoconjugate in the olfactory and respiratory mucosa following inhalation of formaldehyde, respectively.

Componentes glicosilados de la envoltura de Mycobacterium tuberculosis que intervienen en la patogénesis de la tuberculosis / Structural components of the envelope of Mycobacterium tuberculosis that intervene in the pathogenesis of tuberculosis

Los componentes glicosilados de la envoltura de Mycobacterium tuberculosis tienen un papel importante en la inmunopatogénesis de la tuberculosis. Permiten la adhesión, penetración y persistencia de la micobacteria en el macrófago; de igual manera, participan en los mecanismos de activación de estas células y la producción de citocinas relevantes durante la respuesta inmune. En esta revisión, examinamos las características de las principales estructuras sacarídicas de la superficie de la micobacteria y su relación con la modulación de la respuesta inmune.

The glycosylated compounds of Mycobacterium tuberculosis envelope play an important role in the immunopathogenesis of tuberculosis. These molecules are involved in the binding to the host cell surface followed by their internalization and persistence in macrophages; likewise they take part in the macrophage's activation and the production of cytokines that are relevant during the immune response against mycobacteria. In this review we examine the molecular characteristics of the main mycobacterial cell surface saccharide structures and their relation with the modulation of the immune response to tuberculosis.

SO2 Effects on Glycoconjugates of the Respiratory Mucosa in Rat Nasal Cavity / 체질인류학회지

The experiments of this study was performed to investigate the effects of sulfur dioxide on the changes of glycoconjugates of respiratory system of the rat. Sprague -Dawley male rats weighing about 200 ~250g were divided into a control group and SO2 exposed groups. Again SO2 exposed groups were divided into 10 ppm, 25 ppm, 50 ppm, 100 ppm and 200 ppm subgroups according to concentrations of SO2 and each SO2 exposed groups were divided into 1, 3 and 6 hours groups. For the histological changes, H -E(hematoxylin -eosin) and PAS(periodic acid Schiff) staining were used and to investigate the change of sugar residues of glycoconjugates, biotinylated lectins(DBA, SBA, PNA, BSL -1, sWGA, UEA -1, LCA and Con A) were applied. Generally, the effects of SO2 on the rat nasal respiratory region were more serious at the high concentrations. Moreover, as the exposed time was longer even at the low concentrations, the effects of SO2 were similar to those of high concentration. Compared with all SO2 concentrations, the longer exposed time was, the more serious the effects of SO2 were. In the SO2 exposed groups the binding of PNA, RCA -1 and UEA -1 of cilia in the nasal septal respiratory epithelium tended to increase in the 10 ppm and 25 ppm SO2 exposed groups but it tended to decrease in the 100 ppm and 200 ppm SO2 exposed groups. In the cytoplasm of columnar cells of nasal septal respiratory epithelium, Con A binding increased in all the SO2 exposed groups. In the goblet cells DBA, SBA, PNA, RCA -1 and UEA -1 binding increased remarkably in the 50 ppm SO2 exposed groups but it decreased largely or disappeared in the 100 ppm and 200 ppm SO2 exposed groups. The binding of SBA, PNA, BSL -1, UEA -1 and Con A in the intraepithelial mucous cells which were not detected in the control group, increased in the 25 ppm and 50 ppm SO2 exposed groups while it tended to decrease in the 100 ppm and 200 ppm SO2 exposed groups. The binding of sWGA increased according to the concentrations of SO2 were higher and exposed times were longer. In the superior nasal septal gland, the binding of PNA increased in the 50 ppm and 100 ppm SO2 exposed groups and that of Con A increased in the 25 ppm and 50 ppm SO2 exposed groups. In the inferior nasal septal gland, except for LCA, the binding of the other lectins increased remarkably in the 25 ppm and 50 ppm SO2 exposed groups but it tended to decrease in the 100 ppm and 200 ppm SO2 groups. In the mucous duct cells, the reaction of PNA and RCA -1 increased compared with that of the control group. And the reaction of BSL -1 and UEA -1 increased in the lower concentrations of 50 ppm SO2 exposed group but it decreased in the 100 ppm and 200 ppm SO2 exposed groups. The binding of Con A increased in the 25 ppm and 50 ppm SO2 exposed groups. Consequently, from the results above mentioned that SO2 affected serious changes on glycoconjugates metabolism in the nasal cavity.

Study on the effect of products of neuraminidase gene(nan A) in the infection of Streptococcus pneumoniae in chinchillas / 中华传染病杂志

To explore the effect of products of nan A gene on the changes of cell surface carbohydrates of the chinchilla eustachian tube after infection with Streptococcus pneumoniae. Methods Using lectin histochemical techique to compare the changes of the cell surface carbohydrates in the chinchilla eustachian tube after infection with S.pneumoniae D39 or ?NA1 mutant. Results The labeling pattern revealed that the staining with Limax flavus agglutinin (LFA) and Sambucus nigra agglutinin (SNA) was decreased in epithelium of the eustachian tube in the D39 cohort compared to the uninfected control, which indicated that the normal terminal sialic acid residue were removed. Concurrently, the increased staining with wheat germ agglutinin (WGA), succinylated wheat germ agglutinin (Succ WGA), Bandeiraea simplicfolia lectin II (BSL II), peanut agglutinin (PNA) and Erythrina cristagalli lectin (ECL) was observed in the lumen surface of eustachian tube subsequent to intranasal inoculation with D39. However, the ?NA1 neuraminidase deficient mutant did not show any significant changes in the lectin labeling patterns as compared with those of the control cohort. Conclusions The products of the nan A gene play an important role in the changes of cell surface carbohydrates and thus may be responsible for the colonization in the chinchilla eustachian tube after infection with Streptococcus pneumoniae.

Histochemistry on the Effects of Sulfur Dioxide on Glycoconjugates of Rat Skin / 체질인류학회지

This experiment was performed the effect of the glycoconjugates of the skin in the Sprague-Dawley male rats exposed to SO2 for 1, 3 and 6 hours with various concentrations (10, 25, 50, 100 and 200 ppm). To investigate the change of glycoconjugates of skin, seven biotinylated lectins (DBA, SBA, PNA, BSL-1, sWGA, UEA-1, Con A) were applied with ABC method. The epidermis of experiment rats was more or less different according to the concentration of SO2, comparing control group, and beta-N-acetyl-D-galactosamine, alpha-L-fucose, alpha-D-mannose and alpha-D-glucose of the epidermis tend to be increased according to exposed SO2 time, but alpha-D-galactose was tend to be decreased. Galactose-beta-1, 3-Nacetyl- D-galactosamine was tend to more or less increase in horny layer of the epidermis, but decrease in granular and spinous layers. As contrasted control group, beta-N-acetyl-D-glucosamine in the dermis of experiment rats was decreased, but alpha- D-mannose and alpha-D-glucose were increased. In the hair follicle of the experiment rats, beta-N-acetyl-D-galactosamine and galactose-beta-1, 3-N-acetyl-Dgalactosamine tend to be decreased according to exposed SO2 time though more or less differ from the portions of the hair follicle, and beta-N-acetyl-D-glucosamine and alpha-L-fucose were found noticeably to increase comaring control group. According to exposed SO2 time, the decrease of alpha-D-galactose tend to be little more pronounced in the outer root sheath of the upper portion of hair follicle. In contrast, the same sheath of the upper portion in above follicle was increased relatively. alpha-D-mannose and alpha-D-glucose were increased in the root sheath of hair follicle and hair cortex, but decreased in the hair medulla. In the experiment rats, the beta-N-acetyl-D-galactosamine tend to be decreased gradually according to exposed SO2 time, but the decrease of galactose-beta-1, 3-N-acetyl-D-galactosamine were showed significantly as contrasted control group. beta-N-acetyl-D-glucosamine, alpha-D-mannose and alpha-D-glucose were increased gradually according to exposed SO2 time, but alpha-L-fucose was increased remarkably. The effect on lectin binding pattern of the glycoconjugates in the rat skin according to the concentrations and exposed time of SO2 was noted that the alternations of lectin binding pattern tend to be a little more pronounced in low concentration of exposed SO2 for a long time than in high concentration of exposed SO2 for a short time. The alternations of lectin binding pattern were appeared almost similar in concentrations of 50 ppm SO2 and above.

Lectin Histochemistry for Studying Glycoconjugates in Rat Lingual Salivary Glands during the Postnatal Development / 체질인류학회지

The developmental changes of the lingual salivary glands in the postnatal rats were examined by lectin histochemical methods. For the morphological changes, H-E and PAS staining were used. The biotinylated lectins used in the study were DBA, SBA, PNA, BSL-1, sWGA, RCA-1, UEA-1, Con A and LCA. The promordia and undifferentiated acini of the lingual glands were found in the mucous glands at 0 day suckling rat and the von Ebner's glands at 3 day suckling rat, respectively. The differentiation and maturation of the lingual glands were faster than those of the von Ebner's gland. The differentiation and proliferation of both glands were occurred remarkably at suckling periods rather than weaning periods. The lectin binding pattern of glandular promordia and undifferentiated serous acini in von Ebner's gland was weak in BSL-1 and weak to moderate in RCA-1. DBA and sWGA showed tendency to increase in 1 week suckling rat, but The binding reactivity of other lectins was disappeared except BSL-1 that was reacted tracely in 2 and 3 weak suckling and 4 week weaning rat. RCA-1, PNA, sWGA, BSL-1 and SBA of the differentiated serous acini were appeared in the 2 week suckling rat and SBA and sWGA was more intense. Especially, the reactivity of these lectins of suckling periods was showed more tendency to increase than that of weaning periods. The increase of PNA, SBA and BSL-1 was prominent during suckling and weaning periods. RCA-1 and sWGA were decreased in 5 week rat, increased in 6 week rat, and then decreased in adult rat. UEA-1 which was not shown from 0 day to 2 week was showed trace to moderate reactivity in some serous acini. Con A and PNA of glandular promordia and undifferentiated mucous acini were appeared trace or weak, and absent at 0 day suckling rat, but PNA reactivity was showed tendency to incerase at 3 day suckling rat. Other lectins of these promordia and acini were not showed reactivity. In the differentiated mucous acini at 0 day suckling rat, all mucous acini were weak to moderate with DBA, and some of mucous acini also were weak to moderate with BSL-1. Most mucous acini showed weak reactivity with SBA, but some mucous acini showed trace or weak reactivity with RCA, PNA, sWGA and BSL-1. The reactivity of BSL-1 and sWGA was increased from birth to 2 week and then decreased, and absent at 5 week. But it increased at 6 week. RCA-1 and PNA also increased in the acini up to 1 week. However, PNA reactivity was absent at 5 and 6 week. With RCA-1, the intensity of reactivity was increased. Differentiated mucous acini was reacted to increase with SBA from birth, the intensity was strong in weaning periods rather than suckling period. UEA-1 reactivity was showed to decrease from 1 week to 2 week and moderately increased from 3 week to 5 week, and thereafter decreased. DBA binding pattern was somewhat changed throughout the observation periods but it was predominent.

Prelectin Histochemical Study on Glycoconjugates of Rat Lingual Salivary Glands during the Postnatal Development / 체질인류학회지

The developmental changes of the lingual salivary glands in the postnatal rats were examined by morphological and prelectin histochemical methods. For the morphological changes, H -E and PAS staining were used. The mucosubstances stained with PAS, AB pH 2.5, AB pH 1.0 and AF pH 1.7 -AB pH 2.5. The promodia and undifferentiated acini of the lingual glands were detected the mucous glands at o day suckling rat, the von Ebner 's glands at 3 day suckling rat, respectively. The differentiation and maturation of the lingual glands were appeared rapidly than that of the von Ebner 's gland, and that of both glands were occurred remarkably suckling periods more than weaning periods. von Ebner 's gland contained only neutral mucins during postnatal developmental rat. The undifferentiated serous acini of these gland contained a small amounts of neutral mucins from 3 day suckling rat, and the amounts of these mucins were increased continuously according to the differentiation of these glands. The amounts of these mucins predominated at weaning periods more than suckling periods. The lingual mucous glands contained the mixture of neutral and acid mucins, and the amounts of these mucins were tended continuously to increase according to the glandular differentiation. In these glands, the suckling periods were predominant with neutral mucins, but weaning periods were abundant in acid mucins. The differentiated mucous acini of the lingual gland contained large amounts of neutral mucins and small to moderate amounts of acid mucins from 0 day suckling rat, but the undifferentiated mucous acini contained small to moderate amounts of neutral mucins and trace amounts or none of acid mucins in the suckling rat. The amounts of sulfomucin and sialomucin of the differentiated mucous acini was increased in both suckling and weaning periods. The increase of these mucins was markable in weaning periods than suckling periods. The amount of sialomucin was abundant at 0 day suckling rat, but the amount of sulfomucin very increased after 3 day suckling rat, and these mucins strikingly increased after 2 weeks suckling rat.

Lectin Histochemistry for Effects of N - Nitrosodimethylamine on Glycoconjugates in the Rat Lingual Salivary Glands / 체질인류학회지

The effect of NDMA after oral administration (17 mg/ml) on the glycoconjugates of lingual von Ebner's gland and mucous gland were investigated with lectin histochemical methods. For lectin histochemical studies, the biotinylated lectins (DBA, PNA, SBA, BSL -1, sWGA, RCA -1, LCA, UEA -1, and ConA) were applied. Lectin binding patterns of glycoconjugates of lingual von Ebner's gland showed the decreased affinity for DBA, PNA, BSL -1 and sWGA in NDMA -treated group compared with control group. The remarkable decrease of binding affinity of NDMA -treated group was observed in PNA for 12 and 24 hours, DBA for 96 hours, BSL -1 for 72 hours, and sWGA for 3 hours, while the striking decrease of BSL -1 and sWGA binding was observed in NDMA -treated group for 12 hours. But these decreases of binding were tended to recover in PNA and sWGA after 72 hours of NDMA treatment, and in DBA after 120 hours. The binding affinity of SBA and RCA -1 was decreased in NDMA -treated group for 3 hours, while the other NDMA -treated group showed an increased affinity. Especially, the increase of SBA binding was remarkable. There was a little change in binding affinity of UEA -1, LCA and Con A in NDMA -treated group. Lectin binding patterns of glycoconjugates of lingual mucous gland showed decreased affinities for SBA, sWGA and UEA -1 in NDMA -treated group. The striking decreases of binding affinity for NDMA -treated group was observed in SBA and sWGA for 3 hours, and UEA -1 for 3 and 24 hours. And the remarkable decreases of binding affinity for NDMA -treated group was found in SBA for 24 and 48 hours, sWGA for 48, 72 and 96 hours, and UEA -1 for 48 hours. These decreases of binding affinity of NDMA -treated group were tended to recover in SBA and UEA -1 after 96 hours and in sWGA after 120 hours. The binding affinity for PNA and ConA showed a little but not remarkable increase in NDMA - treated group, and LCA binding showed a little decrease following a little increase in NDMA - treated group. The affinity of DBA binding was decreased in NDMA -treated group for 12 hours and 24 hours, while the other NDMA -treated group showed an increased affinity. Especially, there was a remarkable increase in NDMA -treated group for 96 hours. From these results, it is suggested that the toxicity of NDMA may be related with the carcinogen of the rat tongue, and glycoconjugates are concerned with the repaire of the destruction of the lingual mucous acini.

Characterization of the Multinucleated Giant Cell Glycoconjugates in the Tubercle of Human Pulmonary Tuberculosis / 체질인류학회지

Multinucleated giant cells (MGCs) are a prominent characteristic of granulomatous inflammation including tuberculosis. The present study was performed to investigate the characteristics and distribution pattern of intracellular and cell surface glycoconjugates of the MGCs in human pulmonary tubercles using lectin histochemistry. The cytoplasmic staining patterns could be divided into three groups. First, VVL, LCA and SBA showed intense reactivity in the great majority of the MGCs. Second, BS -I, DBA, WGA, PNA, ECL, PHA -L and PHA -E also showed positive staining in the cytoplasm of many MGCs, but the reaction patterns were not uniform. Third, the other group (BS -I - B4 and UEA -I) exhibited very weak or no staining in the cytoplasm. With regard to the membranous staining, the lectin binding patterns could be divided into two groups. First, WGA, ECL, PHA -L & PHA -E showed intense membranous staining. Second, the other lectins (BS -I, BS -I -B4, DBA, VVL, LCA, PNA, SBA and UEA -I) did not show any membranous staining. There was no significant difference in lectin binding patterns between the two types of MGCs. Our results demonstrated the characterization of glycoconjugates expressed in the MGCs in human pulmonary tubercles.

Glycoconjugate Histochemistry of Rat Vaginal Epithelium During the Estrous Cycle / 체질인류학회지

Estrous cycle -related histological and histochemical changes in the vaginal epithelium of mature female rats were studied with PAS (periodic acid Schiff) alcian blue pH 2.5 and biotinylated lectins (DBA, SBA, PNA, BSL -1, sWGA, UEA -1, RCA -1, Con A and LCA).The prominent characteristic changes that occured during the estrous cycle were mucinous transformation in proestrus and cornification in estrus. In proestrus, the superficial mucinous cells of the epithelium were increased in number and enlarged in size, and the amount of acid and neutral mucosubstances was more increase in proestrus than in diestrus and metestrus. About the binding pattern of all lectins examined to the superficial mucinous cells, in diestrus, the binding pattern of these cells showed a similar affinity as in metestrus with intense DBA and UEA -1 reactivity. In proestrus, however, these cells were reactive with seven lectins examined except LCA and PNA, and DBA, SBA, BSL -1, RCA -1 and UEA -1 reacted more strongly than in diestrus and metestrus. In estrus, the superficial cornified cell layers showed a weak reactivity of SBA, BSL -1 and PNA. In diestrus and metestrus, the mucinous cells in the intermediate layers of the basal portion of vaginal fold stained with eight lectins examined except LCA and showed the same binding pattern to the superficial mucinous cells. About the distribution of glycoconjugates in the intermediate layer, the upper spindle cells showed different binding pattern according to the estrous stages. In diestrus, estrus, and metestrus, these cells showed a affinity for all lectins examined. In proestrus, however, DBA and PNA staining were not observed, and stained more intensely with sWGA, SBA and UEA -1, and less intensely with BSL -1 and RCA - 1. In estrus, DBA and PNA reactivity reappeared as trace, and RCA -1 and sWGA reactivity increased. In metestrus, sWGA reactivity reduced and BSL -1 and UEA -1 increased continually. The lower rounded cells of the intermediate layers stained with all lectins examined in estrus, with six lectins examined except Con A, DBA and UEA -1 in proestrus and with five lectins examined except DBA, UEA -1, sWGA and BSL -1 in diestrus and metestrus. BSL -1 reactivity for the layers increased in proestrus, estrus and metestrus, and PNA reactivity increased in estrus and reduced in metestrus. The basal layer of the vaginal epithelium showed different binding pattern to the different portion of vagina, and showed faint staining of BSL -1, SBA and RCA -1, and moderately staining of BSL -1 in proestrus and estrus. In conclusion, alpha /-N -acetyl -D -galactosamine, alpha /-D -galactose and alpha -L -fucose participate in the mucinous transformation of the vaginal epithelium, and beta -N -acetyl -D -glucosamine participates in the cornification of the vaginal epithelium.

Classification of the Carbohydrate using pectin Binding Characteristics and PAS Reaction in Human Gingiva / 체질인류학회지

This study was performed in order to recognize the identifications of the glycoproteins containing oligosaccharides in human gingiva. After made paraffin sections of human gingiva at 4µm, the sections were incubated with 7 lectins (UEA-I, BS-I, SBA, DBA, WGA, PNA, PNA after neuraminidase treated, Con-A). In order to increase specificity of reactions, the sections were applicated with ABC system. And then the sections were incubated with DAB and were counterstained with hematoxylin. Using the same sections, the sections were done H-E and PAS stains. In WGA, DBA and Con-A, plasma membranes of the layers of all epithelium and connective tissue were stained. In BS-I ; In the epithelium of marginal gingiva, plasma membranes of upper layer of the spinous cell layer and granular cell layer were stained. And in epithelium of sulcular gingiva, plasma membranes of the all spinous cell layer and granular cell layer were stained. In SBA ; Plasma membranes of the granular cell layer were stained. In PNA ; In the epithelium of marginal gingiva, plasma membranes of the basal cell layer and lower layer of spinous cell layer were stained. But lectin reactions were not occurred in thc sulcular gingiva. In PNA treated neuraminidase, plasma membranes of the all epithelial layer except basal cell layer membranes especially cytoplasms of upper layer at the sulcular gingiva and connective tissue were reacted. 1. By the above results, authors could know the identification of oligosaccharides existing g1ycoproteins in the human gingiva. 1) All epithelial layer ; α-D-N-Acetyl-Galactosamine, Sialic acid, D-Glucosamine, α-D-Mannose 2) Basal cell layer ; Galactose-β-(1-3)-N-Acetyl-Galactosamine 3) Spinous cell layer ; α-D-Galactose, Galactose-β-(1-3)-N-Acetyl-Galactosamine 4) Granular cell layer ; α-D-Galactose 5) Connective tissue ; α-D-N-Acetyl-Galactosamine, Siallic acid, β-(1-4)-D-Acetyl-Glucosamine, α-D-Glucosamine, α-D-Mannose 2. The Galactose-β-(1-3)-N-Acetyl-Galactosamine was not existed in the basal cell layer and spinous cell layer in the sulcular gingiva.