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Biomedical subjects

W van Dijk

Publications and source records attributed to W van Dijk.

At least 37 records · Page 2Linked to original sources

Pituitary gonadotrophs are strongly activated at the beginning of spermatogenesis in African catfish, Clarias gariepinus.

Pituitary gonadotrophs were studied in male African catfish between 1 and 37 wk of age using antisera against the LH subunits for immunohistological and radioimmunological purposes, and cRNA probes for in situ hybridization. Immunoreactive material was already detectable at the earliest age examined. In juveniles, the signal for the common glycoprotein alpha subunit (GP alpha) was stronger than that for the LH beta subunit. Accordingly, an excess of radioimmunoassayable GP alpha 100 times that of LHbeta was recorded in the pituitary. Using in situ hybridization, the mRNAs were detected 7 (GP alpha) and 13 (LHbeta) wk after hatching. Detection of LHbeta mRNA coincided with a 300-fold increase in the pituitary content of LHbeta and intact LH, whereas GP alpha increased only 15-fold. The number of gonadotrophs per pituitary and the amount of LH per gonadotroph also increased strongly. The strong, initial increase in pituitary LH levels was always associated with the presence of spermatocytes. However, in a limited number of cases (3 out of 12 fish), the pituitary LH content was low despite the presence of spermatocytes. The number of gonadotrophs, the staining intensities (reflecting protein and mRNA), and the pituitary LH content kept increasing, although at a reduced rate, until completion of the first wave of spermatogenesis. In view of the excess of GP alpha over LHbeta, we conclude that expression of the two subunits is regulated in part by different mechanisms, and that expression of LHbeta is rate-limiting for the amount of intact LH. The strong activation of the gonadotrophs shortly after meiosis opens the possibility that a signal of testicular origin stimulates LH expression, in particular its beta subunit. In the absence of a FSH-like gonadotropin in catfish, we propose that LH covers all functions requiring gonadotropic regulation in the African catfish.

Animals↗

Glycosylation of alpha 1-acid glycoprotein in septic shock: changes in degree of branching and in expression of sialyl Lewis(x) groups.

The occurrence of differences in acute-phase response, with respect to concentration and glycosylation of alpha 1-acid glycoprotein (AGP) was studied in the sera of patients, surviving or not from septic shock. Crossed affino-immunoelectrophoresis was used with concanavalin A and Aleuria aurantia lectin for the detection of the degree of branching and fucosylation, respectively, and the monoclonal CSLEX-1 for the detection of sialyl Lewisx (SLeX) groups on AGP. Septic shock apparently induced an acute-phase response as indicated by the increased serum levels and changed glycosylation of AGP. In the survivor group a transient increase in diantennary glycan content was accompanied by a gradually increasing fucosylation and SLeX expression, comparable to those observed in the early phase of an acute-inflammatory response. Remarkably, in the non-survivor group a modest increase in diantennary glycan content was accompanied by a strong elevation of the fucosylation of AGP and the expression of SLeX groups on AGP, typical for the late phase of an acute-phase response. Our results suggest that these changes in glycosylation of AGP can have a prognostic value for the outcome of septic shock.

Acute-Phase Reaction↗

Elevated serum level and altered glycosylation of alpha 1-acid glycoprotein in hyperimmunoglobulinemia D and periodic fever syndrome: evidence for persistent inflammation.

Crossed affinoimmunoelectrophoresis using concanavalin A and Aleuria aurantia lectin as diantennary glycan- and fucose-specific affinocomponents, respectively, was applied to study changes in the concentration and glycosylation of the acute phase protein alpha 1-acid glycoprotein (AGP) in sera obtained from patients with hyperimmunoglobulinemia D and periodic fever syndrome. Increases in concentration of AGP compared to control values were found not only during attacks, but also during remissions. Compared to healthy controls, the presence of diantennary glycan-containing glycoforms of AGP also increased during febrile attacks, while no changes were found during remissions. A continuous high degree of alpha 1-->3 fucosylation was accompanied by a continuous high expression of sialyl Lewisx on AGP. Despite the clinical picture of recurrent febrile attacks with asymptomatic intervals, these studies indicate that hyperimmunoglobulinemia D should be considered a condition of persistent inflammation.

Adolescent↗

Alpha 1-acid glycoprotein (orosomucoid): pathophysiological changes in glycosylation in relation to its function.

The aim of this review is to summarize the research efforts of the last two decades with respect to (i) the determination and characterization of the changes in glycosylation of AGP under various physiological and pathological states; and (ii) the effects of such changes on its possible anti-inflammatory functions. It will become clear that the heterogeneity observed in the glycosylation of AGP in serum, represents various so-called glycoforms of AGP, of which the relative amounts are strictly determined by the (patho) physiological conditions.

Acute Disease↗

Lol p XI, a new major grass pollen allergen, is a member of a family of soybean trypsin inhibitor-related proteins.

BACKGROUND: Monoclonal antibodies were obtained against an unknown allergen from Lolium perenne grass pollen. The allergen had an apparent molecular mass of 18 kd on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Earlier immunoblotting studies had shown that carbohydrate-specific IgG antibodies recognize an antigen of similar size. OBJECTIVE: We sought to characterize the allergen biochemically and immunologically. METHODS: The amino acid sequence of the allergen was determined by automated Edman degradation, and its monosaccharide composition was determined by gas chromatographic analysis. A panel of 270 grass pollen-positive sera was assessed in a RAST with the purified allergen. Protease digestion (proteinase K) and chemical deglycosylation (trifluoromethane sulfonic acid) were used to distinguish between carbohydrate and peptide epitopes for IgE antibodies. RESULTS: The allergen was shown to be a glycoprotein with a molecular mass of 16 kd, of which 8% is carbohydrate. Its amino acid sequence shares 32% homology with soybean trypsin inhibitor (Kunitz) but lacks its active site. No homology was found with known grass pollen allergens, hence it was designated Lol p XI. A high degree of homology (44%) was found with a tree pollen allergen, Ole e I, the major allergen of olive pollen. More than 65% of grass pollen-positive sera had IgE against Lol p XI. IgE reactivity was demonstrated both with the carbohydrate moiety and the peptide backbone. CONCLUSIONS: Lol p XI is a new major grass pollen allergen carrying an IgE-binding carbohydrate determinant. Lol p XI is structurally related to the major allergen from olive pollen.

Allergens↗

Pubertal development of male African catfish (Clarias gariepinus). Pituitary ultrastructure and responsiveness to gonadotropin-releasing hormone.

Pubertal development was studied in male African catfish by immunocytochemical examination of pituitary gonadotrophs and by monitoring the responsiveness of gonadotropin (GTH) secretion to salmon GnRH analogue (sGnRHa) in vitro. Experiments were carried out with fish from 9 to 28 wk of age. Fish were assigned to four groups, according to the stage of spermatogenesis: I, spermatogonia alone; II, spermatogonia and spermatocytes; III, spermatogonia, spermatocytes, and spermatids; IV, all germ cell stages, including spermatozoa. Basal and sGnRHa-stimulated secretion of the LH-like GTH II increased 3- to 4-fold from stage I to II and from stage II to III, whereas a 15-fold increase was recorded from stage III to IV. The ED50 values of sGnRHa varied between 0.08 and 0.49 nM, stages II and III being less sensitive. The highest dosage of sGnRHa (100 nM) led to a reduction of GTH secretion. In the first three stages, the pituitary secreted large amounts of free alpha-subunit while free GTH II beta-subunit was not detected at any stage of development. Antisera against GTH II and its alpha- and beta-subunits were used for immunocytochemical studies. In stages I and II, two subtypes of gonadotrophs, which differed in the size and labeling intensity of their secretory granules, were present. Both types of granules were immunopositive for the two subunits of GTH II. In stages III and IV, only gonadotrophs of the subtype with the larger granules were found. Globules and irregular, membrane-bound masses (IMs), probably arising through fusion of secretory granules, appeared in the gonadotrophs in stage III and became more prominent in stage IV. Globules and IMs were immunopositive for the beta-subunit but negative for the alpha-subunit. We conclude that the two subtypes of gonadotrophs represent different developmental stages of GTH II-producing cells, as they shared immunolabeling for the alpha- and the beta-subunits of GTH II. The scarcity of GTH II beta-subunit may be rate-limiting for the amount of intact GTH II available for secretion, particularly at early stages of development. In contrast, at more advanced stages when the readily releasable pool of GTH II has greatly increased, the amount of GTH II also appears to be controlled by modification or elimination of the alpha-subunit from globules and IMs.

Amino Acid Sequence↗

Antimetabolite-induced increases in the invasive capacity of murine leukaemia L1210 cells.

Pretreatment of murine leukaemia L1210 cells with non-lethal concentrations of various antimetabolites increased the in vitro invasive capacity of these cells into monolayers of rat embryo fibroblasts. The increase in invasive capacity was partly correlated with the induced cell cycle arrest. The concomitant increase in cell surface fucosylation and inhibition of invasion with sulphate indicate a role for glycoproteins in this process. Our results suggest that treatment with antimetabolites may lead to a more aggressive phenotype by altering cell surface properties.

Animals↗

Diagnosis of hepatitis C virus infection using two second-generation enzyme immunoassays with a recombinant immunoblot assay for confirmation.

A total of 1,016 serum samples from patients with either non-A, non-B hepatitis or risk factors for hepatitis C virus (HCV) infection were examined in two second-generation enzyme immunoassays (EIAs), the UBI HCV EIA (Organon Teknika, The Netherlands) and the Wellcozyme anti-HCV (Murex Diagnostics, UK), for detection of antibodies to HCV. An immunoblot assay that uses four recombinant antigens, the 4-RIBA (Chiron, USA), was used as a confirmatory assay. Of the 1,016 samples, 195 (19.2%) were reactive in both EIAs, while ten yielded discrepant results. One hundred eighty of the 195 (92%) positive reactions were confirmed in the 4-RIBA; 13 sera yielded an indeterminate result and two were negative. None of the sera with discrepant results reacted positively in the confirmatory test, while two sera showed an indeterminate pattern. In contrast to the screening of antibodies to HCV among blood donors, confirmatory testing of antibodies to HCV with the 4-RIBA seems to have limited added value in the diagnostic examination of clinical samples from patients with suspected HCV infection.

Diagnostic Errors↗

Heterogeneity of human biliary mucin: functional implications.

1. Human gallbladder mucin has been implicated as playing a role in the pathogenesis of gallstones. In previous studies no differences have been found in the content or composition of mucins derived from control bile or cholesterol gallstone bile. Until now, no differences were also found between these two groups of mucins with regard to their ability to cause cholesterol nucleation. In the accompanying paper we have reported that there is a strong heterogeneity of gallbladder mucins derived from individual patients (M. J. A. van Wijland, J. H. Klinkspoor, L. Th. de Wit, R. P. J. Oude Elferink, G. N. J. Tytgat and A. K. Groen, Clin Sci 1994; 86: 67-74). In the present study we further investigated a possible patient to patient heterogeneity of mucin by means of immunological and functional characterization of mucins isolated from hepatic bile of six different patients with gallstones. 2. Considerable heterogeneity was found. Two of the mucins barely reacted with a polyclonal anti-mucin antibody, whereas the other four mucins reacted very strongly. Lectin-binding studies indicated that the glycans of these two mucins expressed less D-N-acetylgalactosamine residues than the other four mucins. This was confirmed by analyses of the glycan compositions. These studies furthermore indicated that the glycans were of the O-linked type, contained alpha-D-N-acetylglucosamine and were fucosylated, sialylated and sulphated to different extents. Except for a strong heterogeneity in the sugar composition of the mucins, heterogeneity was also found in the biological activity of the mucins.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

Inflammation-induced expression of sialyl Lewis X-containing glycan structures on alpha 1-acid glycoprotein (orosomucoid) in human sera.

The glycosylation of the acute phase glycoprotein alpha 1-acid glycoprotein (AGP) in human sera is subject to marked changes during acute inflammation as a result of the cytokine-induced hepatic acute phase reaction. The changes described thus far comprise alterations in the type of branching of the carbohydrate structures as revealed by increased reactivity of AGP with concanavalin A. We now report on acute inflammation-induced increases in alpha 1-->3-fucosylated AGP molecules, as detected by the reactivity of AGP towards the fucose-binding Aleuria aurantia lectin (AAL) in crossed affino-immunoelectrophoresis of human sera. Laparotomy of women, for the removal of benign tumors of the uterus, was used as a model for the development of the hepatic acute phase response. Hugh increases were detected in the amounts of strongly AAL-reactive fractions of AGP, presumably containing three or more fucosylated N-acetyllactosamine units. At least part of these Lewis X-type glycans (Gal beta 1-->[Fuc alpha 1-->3]GlcNAc-R) appeared to be substituted also with an alpha 2-->3-linked sialic acid residue. This was revealed by the laparotomy-induced abundant staining of AGP with an antisialyl Lewis X monoclonal antibody (CSLEX-1) on blots of sodium dodecyl sulfate-polyacrylamide gels containing AGP isolated from the sera of a patient at various days after operation. It is concluded that acute inflammation induces a strong increase in sialyl Lewis X-substituted AGP molecules that persists at a high level throughout the inflammatory period. We postulate that these changes represent a physiological feedback response on the interaction between leukocytes and inflamed endothelium, which is mediated via sialylated Lewis X structures and the selectin endothelial-leukocyte adhesion molecule 1.

Acute-Phase Proteins↗

The utility of lipid-associated sialic acid (LASA or LSA) as a serum marker for malignancy. A review of the literature.

The utility of the lipid-associated sialic acid (LASA or LSA) test as a serum marker for malignancy is reviewed. The name LASA or LSA test is confusing because it suggests that only or mainly lipid-bound sialic acid is measured. In reality, glycoprotein-bound sialic acid is determined predominantly. The assay appears to have a particularly high positivity rate in leukemia, Hodgkin's disease, melanoma, sarcoma, advanced ovarian carcinoma and oropharyngeal tumors, suggesting that LASA may serve as a valuable marker in these malignancies. As a consequence of the rise of sialic acid-rich acute-phase proteins, such as alpha 1-acid glycoprotein, in inflammatory diseases the specificity of LASA and therefore its diagnostic accuracy is low. LASA can be useful for monitoring cancer patients during treatment, especially in combination with other tumor markers.

Antigens, Neoplasm↗

Inflammation-induced changes in expression and glycosylation of genetic variants of alpha 1-acid glycoprotein. Studies with human sera, primary cultures of human hepatocytes and transgenic mice.

The relative occurrence of genetic variants of human alpha 1-acid glycoprotein (AGP) in relation to changes in glycosylation was studied in sera of patients with burn injury, media of cytokine-treated primary cultures of human hepatocytes and Hep 3B cells, and sera of transgenic mice expressing the human AGP-A gene. It is concluded (i) that the glycosylation of AGP was not dependent on its genetic expression and (ii) that both the variants determined by the AGP-A gene as well as by the AGP-B/B' genes are increased after inflammation or treatment with interleukins 1 and 6.

Animals↗

Determining the 222Rn exhalation rate of building materials using liquid scintillation counting.

A new method for determining the free 222Rn exhalation rate from building materials is described. The sample is enclosed in a container from which the exhaled Rn is continuously purged by nitrogen gas. After 2-3 h, when the Rn level in the container has reached a steady-state concentration, the outflowing Rn is trapped on silica gel at about -190 degrees C. About 16 h after sampling, the silica gel is analyzed by liquid scintillation counting to determine the area exhalation rate. The method described has a good repeatability and reproducibility with coefficients of variation of 7.8% and 8.3%, respectively, at 5 Bq m-2 h-1. The low limit of detection of 11 mBq 222Rn offers the opportunity to quantify the exhalation rate of almost all kinds of building materials. It was found that the air humidity strongly influences the exhalation rates of building material and, therefore, should be controlled. Two typical building materials were investigated. For gypsum, an increase in the exhalation rate with increasing water vapor pressure was found, whereas for concrete, a linear decrease with increasing water vapor pressure was observed. The 222Rn area exhalation rates of 20 Dutch building materials, including some experimental ones, were determined at 50% RH, 20 degrees C, showing a range of less than 0.02-15.8 Bq m-2 h-1. The lowest values were found for natural gypsum board, the highest for phosphogypsum blocks. Building materials containing fly ash gave area exhalation rates comparable to those of similar materials without fly ash.

Air Pollutants, Radioactive↗

Effect of transgenic expression of human alpha 1-acid glycoprotein (AGP) on the glycosylation of human and mouse AGP in various transgenic mouse sera.

The occurrence and the glycosylation of human alpha 1-acid glycoprotein (AGP) was studied in two classes of transgenic mice expressing either the A, B and B' genes (ABB'-mice) or only the A gene of human AGP (A-mice). The glycosylation of the human AGP molecules in the transgenic mouse sera was compared with the glycosylation of mouse AGP in the same animal and with human AGP in normal human serum by studying their heterogeneity in binding to concanavalin A (Con A), using crossed affino immunoelectrophoresis (CAIE) with Con A as the affinocomponent in the first dimension gel. Three to four different glycosylated fractions of human as well as mouse AGP were revealed by this method in all the transgenic mouse sera. A close relationship was apparent between the heterogeneities in Con A binding of human and mouse AGP in the same transgenic mouse. The magnitude of this so-called Con A reactivity was, however, strongly dependent on the transgenic mouse studied. Especially within the group of ABB'-mice dramatic changes in Con A reactivity were found when the human AGP genes were expressed. This indicates in the first place that the oligosaccharide chains of the human AGP molecules expressed also mouse-specific features. Secondly, and more importantly, these findings indicate that the expression of the human AGP genes affected the glycosylation process of the transgenic mouse liver. This organ is the source of the AGP forms occurring in serum. We do not know whether this effect has been caused by the introduction or the expression of the human gene(s) or by the presence of human AGP in the Golgi system or in serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute-Phase Proteins↗

How to become involved in EDS.

This article presents ways in which dental students may become involved in EDS. It is important that EDS has the support of dental students as it is a Committee which works in their interest.

European Union↗

Isolation and characterization of BanLec-I, a mannoside-binding lectin from Musa paradisiac (banana).

A lectin (BanLec-I) from banana (Musa paradisiac) with a binding specificity for oligomannosidic glycans of size classes higher than (Man)6GlcNAc was isolated and purified by affinity chromatography on a Sephadex G-75 column. It did not agglutinate untreated human or sheep erythrocytes, but it did agglutinate rabbit erythrocytes. BanLec-I stimulated T-cell proliferation. On size-exclusion chromatography, BanLec-I has a molecular mass of approx. 27 kDa, and on SDS/PAGE the molecular mass is approx. 13 kDa. The isoelectric point is 7.2-7.5. BanLec-I was found to be very effective as a probe in detecting glycoproteins, e.g. on nitrocellulose blots.

Animals↗