Biomedical subjects

A van der Heijden

Publications and source records attributed to A van der Heijden.

15 recordsLinked to original sources

PubMed2004

Expression and activity of citrullinating peptidylarginine deiminase enzymes in monocytes and macrophages.

BACKGROUND: Antibodies directed to proteins containing the non-standard amino acid citrulline, are extremely specific for rheumatoid arthritis (RA). Peptidylcitrulline can be generated by post-translational conversion of arginine residues. This process, citrullination, is catalysed by a group of calcium dependent peptidylarginine deiminase (PAD) enzymes. OBJECTIVE: To investigate the expression and activity of four isotypes of PAD in peripheral blood and synovial fluid cells of patients with RA. RESULTS: The data presented here show that citrullination of proteins by PAD enzymes is a process regulated at three levels: transcription-in peripheral blood PAD2 and PAD4 mRNAs are expressed predominantly in monocytes; PAD4 mRNA is not detectable in macrophages, translation-translation of PAD2 mRNA is subject to differentiation stage-specific regulation by its 3' UTR, and activation-the PAD proteins are only activated when sufficient Ca(2+) is available. Such high Ca(2+) concentrations are normally not present in living cells. In macrophages, which are abundant in the inflamed RA synovium, vimentin is specifically citrullinated after Ca(2+) influx. CONCLUSION: PAD2 and PAD4 are the most likely candidate PAD isotypes for the citrullination of synovial proteins in RA. Our results indicate that citrullinated vimentin is a candidate autoantigen in RA.

Arthritis, Rheumatoid↗

PubMed2001

Identification of a novel cis-acting RNA element involved in nuclear export of hY RNAs.

Ro RNPs are small cytoplasmic RNA-protein complexes of unknown function that have been found in all metazoan cells studied so far. In human cells, Ro RNPs consist of one of four small RNA molecules, termed hY RNAs and at least two well-characterized proteins, Ro60 and La. In previous Xenopus laevis oocyte microinjection studies, we showed that an intact Ro60 binding site (Stem-loop 1) is a prerequisite for efficient nuclear export of hY1 RNA, whereas an intact La-binding site promotes nuclear retention (Simons et al. RNA, 1996, 2:264-273). Here we present evidence that the distal half (Stem 2) of the conserved base-paired stem structure found in all hY RNAs also plays a critical role in the export process. A minimal RNA molecule containing this region, L1S2 RNA, competes effectively for the export of full-length hY1 RNAs and is itself exported very rapidly in a Ro60-independent and RanGTP-dependent manner. Mutational analyses of this RNA shows that a 5'/3' terminal double-stranded stem structure (>10 bp) of no specific nucleotide sequence constitutes a novel nuclear export element (NEE). Cross-competition studies indicate that this type of NEE may also be involved in export of other classes of RNAs. Like full-length hY1 RNA, L1S2 RNA also competes for export of ET-202 RNA, an RNA that was selected for its efficient nuclear export in the presence of the nuclear transport inhibitor, VSV Matrix protein (Grimm et al. Proc Natl Acad Sci USA, 1997, 94:10122-10127). However, export of L1S2 RNA is strongly inhibited by VSV-M protein, showing that these RNAs use partially overlapping, but not identical export pathways. We propose that export of Y RNAs is mediated by two contiguous cis-acting elements in the 5'/3' double-stranded stem region that is conserved between different Y RNAs.

Active Transport, Cell Nucleus↗

PubMed2001

Basic domains target protein subunits of the RNase MRP complex to the nucleolus independently of complex association.

The RNase MRP and RNase P ribonucleoprotein particles both function as endoribonucleases, have a similar RNA component, and share several protein subunits. RNase MRP has been implicated in pre-rRNA processing and mitochondrial DNA replication, whereas RNase P functions in pre-tRNA processing. Both RNase MRP and RNase P accumulate in the nucleolus of eukaryotic cells. In this report we show that for three protein subunits of the RNase MRP complex (hPop1, hPop4, and Rpp38) basic domains are responsible for their nucleolar accumulation and that they are able to accumulate in the nucleolus independently of their association with the RNase MRP and RNase P complexes. We also show that certain mutants of hPop4 accumulate in the Cajal bodies, suggesting that hPop4 traverses through these bodies to the nucleolus. Furthermore, we characterized a deletion mutant of Rpp38 that preferentially associates with the RNase MRP complex, giving a first clue about the difference in protein composition of the human RNase MRP and RNase P complexes. On the basis of all available data on nucleolar localization sequences, we hypothesize that nucleolar accumulation of proteins containing basic domains proceeds by diffusion and retention rather than by an active transport process. The existence of nucleolar localization sequences is discussed.

Apoptosis Regulatory Proteins↗

PubMed2000

Detailed analysis of the phosphorylation of the human La (SS-B) autoantigen. (De)phosphorylation does not affect its subcellular distribution.

The La (SS-B) autoantigen is an evolutionarily conserved phosphoprotein which plays an important role, most likely as an RNA chaperone, in various processes, such as the biosynthesis and maturation of RNA polymerase III transcripts in the cell nucleus and (internal) initiation of translation in the cytoplasm. In this study, the phosphorylation state of this protein from human HeLa and HEp-2 cells was characterized by high-resolution two-dimensional IEF/SDS-PAGE analysis, and phosphorylation sites were mapped by nanoelectrospray mass spectrometry. Furthermore, the effect of phosphorylation at the sites identified on the subcellular distribution of the protein was studied by site-directed mutagenesis. At least 14 isoelectric isoforms were discerned on 2-D gels with La protein from both types of cells. Metabolic labeling in combination with alkaline phosphatase treatment revealed that only a limited number of these isoforms could be attributed to phosphorylation. Four phosphorylation sites, Thr-302, Ser-325, Thr-362, and Ser-366, were mapped by mass spectrometric analysis of the isolated La protein from HeLa cells or the carboxy-terminal half of this protein. The analysis of mutants of La, in which the respective phosphorylated residues were replaced by either a neutral (alanine) or an acidic (aspartate) residue, by microinjection into Xenopus laevis oocytes on the one hand and transfection of HEp-2 cells on the other hand revealed that the subcellular distribution of this protein was not affected by these amino acid substitutions. These results strongly suggest that the signals that determine the subcellular distribution of this protein are not regulated by (de)phosphorylation of the target residues examined.

Amino Acid Sequence↗

PubMed1999

Rapid nucleolytic degradation of the small cytoplasmic Y RNAs during apoptosis.

We have investigated the fate of the RNA components of small ribonucleoprotein particles in apoptotic cells. We show that the cytoplasmic Ro ribonucleoprotein-associated Y RNAs are specifically and rapidly degraded during apoptosis via a caspase-dependent mechanism. This is the first study describing the selective degradation of a specific class of small structural RNA molecules in apoptotic cells. Cleavage and subsequent truncation of Y RNAs was observed upon exposure of cells to a variety of apoptotic stimuli and were found to be inhibited by Bcl-2, zinc, and several caspase inhibitors. These results indicate that apoptotic degradation of Y RNAs is dependent on caspase activation, which suggests that the nucleolytic activity responsible for hY RNA degradation is activated downstream of the caspase cascade. The Y RNA degradation products remain bound by the Ro60 protein and in part also by the La protein, the only two proteins known to be stably associated with intact Ro ribonucleoprotein particles. The size of the Y RNA degradation products is consistent with the protection from degradation of the most highly conserved region of the Y RNAs by the bound Ro60 and La proteins. Our results indicate that the rapid abrogation of the yet unknown function of Y RNAs might be an early step in the systemic deactivation of the dying cell.

Anisomycin↗

PubMed1999

Trace-level determination of polar flavour compounds in butter by solid-phase extraction and gas chromatography-mass spectrometry.

Volatile compounds are responsible for the aromas of butter. A simple technique for the determination of these components is described which is based on solid-phase extraction (SPE) after melting of the butter and separation of the aqueous phase from the fat. Volatile flavours present in the water fraction are collected by off-line SPE on cartidges packed with a copolymer sorbent. After desorption with 500 microliters of methyl acetate, 1-microliter aliquots are quantified and/or identified by gas chromatography-mass spectrometry. The procedure was tested with respect to recovery, linearity and limit of detection in real-life samples using five polar model analytes. It allows the characterisation of polar flavour compounds in butter prior to and after heat treatment at 170 degrees C. From the five model compounds, vanillin, traces of diacetyl and maltol were found to be present in the butter samples. After heat treatment 500-1000-fold increased concentration of maltol, and substantial amounts of furaneol were detected.

Benzaldehydes↗

PubMed1999

The La (SS-B) autoantigen, a key protein in RNA biogenesis, is dephosphorylated and cleaved early during apoptosis.

In the past few years, a role for apoptotic processes in the development of autoimmune diseases has been suggested. An increasing number of cellular proteins, which are modified during apoptosis, has been described, and many of these proteins have been identified as autoantigens. We have studied the effects of apoptosis on the La protein in more detail and for the first time demonstrate that this autoantigen is rapidly dephosphorylated after the induction of apoptosis. Dephosphorylation of the La protein was observed after induction of apoptosis by several initiators and in various cell types. Furthermore, we demonstrate that at least a subset of the La protein is proteolytically cleaved in vivo, generating a 45 kDa fragment. Dephosphorylation as well as cleavage of La is inhibited by ZnSO4 as well as by several tetrapeptide caspase inhibitors, indicating that these processes require the activation of caspases. Dephosphorylation of La is inhibited by low concentrations of okadaic acid, suggesting that a PP2A-like phosphatase is involved. Generation of the 45 kDa fragment is consistent with proteolytic cleavage at amino acids 371 and/or 374. The possible significance of the apoptotic changes in the La protein for autoantibody production is discussed.

Amino Acid Sequence↗

PubMed1996

Actinomycosis, a sheep in wolves' clothes.

We present a patient referred for radiotherapy for a presumed pulmonary malignancy, who was found to suffer from an actinomycotic infection. This case illustrates the importance of early consideration of actinomycosis when diagnostic methods are negative for malignancy or specific chestwall and bony lesions are observed.

Actinomycosis↗

PubMed1995

Spleen pigmentation in young C57BL mice is caused by accumulation of melanin.

It has been previously reported that in 2 C57BL mouse sublines a dark pigmentation of the cranial part of the spleen occurs in up to 30% of the animals within the populations. It was not clear whether this discoloration is caused by melanosis, lipofuscinosis or haemosiderosis. With the use of light and electron microscopy of stained spleen sections, we identified the pigment in 14 out of 60 C57BL mice aged 8-10 wks. In the mice with pigmented spleens there was accumulation of melanin, predominantly in melanophores. Literature data indicate that apart from melanin, lipofuscin and haemosiderin can be observed in splenic macrophages provided that the mice are older than those studied by us. We conclude that melanin is the principal pigment causing spleen discoloration in young C57BL mice. Splenic melanosis displays inter-individual variation, but its relevance from a pathophysiological point of view remains obscure.

Animals↗

PubMed1994

Dietary fructose v. glucose in rats raises urinary excretion, true absorption and ileal solubility of magnesium but decreases magnesium retention.

Apparent Mg absorption, that is Mg intake minus faecal excretion, was found to be greater in rats fed on diets containing fructose instead of glucose. This effect of fructose was accompanied by enhanced urinary excretion and diminished retention of Mg. True Mg absorption was then determined with the use of oral and intraperitoneal administration of tracer doses of 28 Mg. True Mg absorption was significantly greater in rats fed on fructose. There was no significant effect of fructose v. glucose on faecal excretion of endogenous Mg. It was hypothesized that fructose enhances the solubility of Mg in the ileal lumen and thereby facilitates its absorption. The distribution of Mg between the solid and liquid phases of the ileum was determined in rats fed on either glucose or fructose. Fructose reduced the amount of Mg in the solid phase but raised both the amount and the concentration of Mg in the liquid phase. We conclude that the dietary-fructose-induced stimulation of Mg absorption in rats is caused by a raised solubility of ileal Mg, but the mechanism by which fructose exerts this effect and why it was not associated with a decrease in faecal excretion of endogenous Mg remain unknown. Dietary fructose v. glucose did not systematically affect the apparent absorption of Ca and P.

Animals↗

PubMed1992

Correspondence.

Explore the source record for details and available documents.

Letter↗

PubMed1991

An unusual cause of diplopia in a cancer patient.

A 47-year-old woman with metastatic infiltrating lobular carcinoma of the breast developed diplopia. Computed tomography of the orbits showed enlargement and irregularity of the right inferior rectus and inferior obliques muscles. Biopsies of these muscles contained breast carcinoma cells. This case report discusses the causes of diplopia in cancer patients, with special attention to the diagnostic problems of metastasis in extraocular muscles. The possible combined occurrence of metastasis in the leptomeninges and extraocular muscles is also to be borne in mind if the latter diagnosis is not to be missed.

Breast Neoplasms↗

PubMed1987

Bromocriptine is unable to suppress TRH-stimulated beta-endorphin/beta-lipotropin and cortisol secretion in normal pregnant women after delivery.

The course of plasma beta-endorphin/beta-lipotropin, cortisol and prolactin (PRL) levels was followed from 0.5 till 5 h after normal delivery in 13 healthy women. Six subjects who did not want to breast-feed their child received 2.5 mg bromocriptine orally 1 h after delivery. After 3 h the effect of the intravenous administration of 200 micrograms thyrotropin-releasing hormone (TRH) was also measured. Elevated plasma beta-endorphin and cortisol levels decreased after delivery in a (log) linear fashion which was not influenced by bromocriptine. TRH elicited a significant short-lived identical increase in plasma beta-endorphin/beta-lipotropin concentrations in the control and the bromocriptine-treated subjects. TRH similarly delayed the rapid decline in plasma cortisol levels in both groups of women. Basal and TRH-induced PRL levels were rapidly suppressed by bromocriptine. These studies show the presence of a paradoxical increase of beta-endorphin/beta-lipotropin and cortisol levels in response to TRH occurring shortly after delivery in normal women. This response cannot be mediated by the placenta. The absence of an inhibiting effect of bromocriptine on basal and TRH-induced beta-endorphin and cortisol release does not lend support to the hypothesis of the presence of a functionally active intermediate pituitary lobe in man early in puerperium.

Bromocriptine↗

PubMed1975

Structure-taste relationship of some sweet-tasting dipeptide esters.

The sweetness of dipeptide esters, structurally related to L-aspartyl-L-phenylalanine methyl ester (Aspartame), depends among other things on molecular size. This paper describes a method for predicting whether a dipeptide ester is sweet or not with the aid of atomic models. The method is based on the determination of the size, length and shape of the side chain (R) of the amino-acid ester attached to L-aspartic acid. We measured the side chains of 28 dipeptide esters, 13 of which were synthesized and evaluated for sweetness in our laboratory. Data of the other esters were taken from the literature. Twentyone dipeptide esters were sweet; the size and length of the side chains could be correlated with the degree of sweetness. An ester is sweet provided the length of the side chain (R) is between 4.8 and 8.8 A, and its size greater than or equal to 29 A; as for shape of R in sweet esters, there were two maxima in space filling at about 2 and 4 A or about 3 and 5 A from the asymmetric carbon atom.

Aspartame↗

PubMedDate not supplied

[Effect of increasing protein ingestion on the nitrogen balance of mechanically ventilated critically ill patients receiving total parenteral nutrition].

The amount of protein recommended to minimise N loss in critically ill patients receiving total parenteral nutrition (TPN) varies in the literature. Therefore, we studied the effect of increased protein intake on the N balance, administering TPN with either 1.2 g protein/kg/day (low N diet) or 1.8 g protein/kg/day (high N diet). Fifteen mechanically ventilated critically ill patients were studied in a surgical intensive care unit. After at least two days of standard TPN, patients were randomly assigned to either the low or the high N diet. Ten patients were studied on the low N diet and 11 on the high N diet; 6 patients were studied on both diets. Nonprotein energy was supplied according to estimated energy requirements. For five consecutive days, the N balance was measured daily. Total urinary nitrogen (TUN) was analysed using the Kjeldahl method. There was no difference in N balance between the groups. On the low N diet, N balance was -0.113 +/- 0.088 and on the high N diet -0.113 +/- 0.109 g N/kg/day. In patients studied twice, N balance was -0.087 +/- 0.054 and -0.050 +/- 0.060 g N/kd/day respectively. Results of a previous pilot study showed that in 20 similar patients the N balance became 80% less negative (from -5.7 +/- 5.1 to -1.1 +/- 8.2 g N/day) when protein intake was increased from 0.9 to 1.5 g/kg/day. Since these results are consistent with other studies, we conclude that the optimal range of protein supply in this type of critically ill patients is approximately 1.1-1.5 g protein/kg/day.

Adolescent↗