PubMed Health⌕ Search

Biomedical subjects

A Claeys

Publications and source records attributed to A Claeys.

At least 19 recordsLinked to original sources
PubMed2002

Diplopia as an initial manifestation of disseminated non-hodgkin's lymphoma.

A 37-year-old male presented a sudden diplopia. Ocular examination showed a partial paresis of the left eye (LE) on the left gaze, progressing in a few days towards a total ophthalmoplegia. Further investigation revealed an intra-orbital mass, immunohistologically diagnosed as a Diffuse Large B-cell Lymphoma (DLBCL), according to the WHO classification. Since the patient was in an advanced, disseminated stage of the disease (IVA-E), treatment was based on systemic and intrathecal chemotherapy with a pancranial radiotherapy. The clinical course was poor with only a 10-month survival. We wish to stress that the possibility of orbital malignancy in young adults with acute onset of ophthalmoplegia should be included in the differential diagnosis.

Adult↗
PubMed2000

High-level expression of active HIV-1 integrase from a synthetic gene in human cells.

A synthetic gene encoding for HIV-1 integrase was designed to circumvent the intrinsic instability and the repressor elements present in the wild-type gene. High-level expression of HIV-1 integrase was obtained in various human cell lines independently of viral accessory proteins. A human 293T cell line was selected that stably expresses HIV-1 integrase and has growth kinetics comparable to the parental cell line. The enzyme was localized in the nucleus and remained stably associated with the chromosomes during mitosis. Lentiviral vector particles carrying the inactivating D64V mutation in the integrase gene were capable of stably transducing 293T cells when complemented in the producer cells with integrase expressed from the synthetic gene. When the cell line that stably expresses integrase was infected with the defective viral particles, complementation of integrase activity was detected as well. Expression of active HIV-1 integrase in human cells will facilitate the study of the interplay between host and viral factors during integration.

Amino Acid Sequence↗
PubMed2000

DNA-Dependent protein kinase is not required for efficient lentivirus integration.

How DNA is repaired after retrovirus integration is not well understood. DNA-dependent protein kinase (DNA-PK) is known to play a central role in the repair of double-stranded DNA breaks. Recently, a role for DNA-PK in retroviral DNA integration has been proposed (R. Daniel, R. A. Katz, and A. M. Skalka, Science 284:644-647, 1999). Reduced transduction efficiency and increased cell death by apoptosis were observed upon retrovirus infection of cultured scid cells. We have used a human immunodeficiency virus (HIV) type 1 (HIV-1)-derived lentivirus vector system to further investigate the role of DNA-PK during integration. We measured lentivirus transduction of scid mouse embryonic fibroblasts (MEF) and xrs-5 or xrs-6 cells. These cells are deficient in the catalytic subunit of DNA-PK and in Ku, the DNA-binding subunit of DNA-PK, respectively. At low vector titers, efficient and stable lentivirus transduction was obtained, excluding an essential role for DNA-PK in lentivirus integration. Likewise, the efficiency of transduction of HIV-derived vectors in scid mouse brain was as efficient as that in control mice, without evidence of apoptosis. We observed increased cell death in scid MEF and xrs-5 or xrs-6 cells, but only after transduction with high vector titers (multiplicity of infection [MOI], >1 transducing unit [TU]/cell) and subsequent passage of the transduced cells. At an MOI of <1 TU/cell, however, transduction efficiency was even higher in DNA-PK-deficient cells than in control cells. Taken together, the data suggest a protective role of DNA-PK against cellular toxicity induced by high levels of retrovirus integrase or integration. Another candidate cellular enzyme that has been claimed to play an important role during retrovirus integration is poly(ADP-ribose) polymerase (PARP). However, no inhibition of lentivirus vector-mediated transduction or HIV-1 replication by 3-methoxybenzamide, a known PARP inhibitor, was observed. In conclusion, DNA-PK and PARP are not essential for lentivirus integration.

Animals↗
PubMed1990

Assay for acetyl-CoA:arylamine N-acetyltransferase by high-performance liquid chromatography applied to serotonin N-acetylation.

A specific assay to measure the activity of the enzyme acetyl-CoA:arylamine N-acetyltransferase (EC 2.3.1.5) from pigeon liver is described. The assay is based on the HPLC analysis of N-acetylserotonin formed by the enzymatic reaction. A reversed-phase column (Spherisorb 5-microns ODS 2; 150 x 3.2 mm) eluted with 0.1 M sodium acetate (pH 4.75)/methanol (75:25) permits baseline separation of serotonin and N-acetylserotonin within 5.3 min. Several variables on the enzyme reaction were studied to obtain maximum activity. The enzyme is most active in glycine buffer at pH 9.5. The apparent Km value for serotonin (at 0.6 mM CoASAc) is 0.246 mM and 9.9 microM for CoASAc (at 1.5 mM serotonin). To avoid acetyl-CoA or N-acetylserotonin consumption in side-reactions, the enzyme was purified. A two-step purification process (ammonium sulfate fractionation and affinity chromatography on immobilised amethopterin) yielded 60-70% of the initial enzyme activity with a purification factor of 455-560.

Acetyl Coenzyme A↗
PubMed1990

Automated analysis of muscle fibre images.

A technique for studying the distribution and size of different fibre types in muscles is proposed for automated analysis of individual fibres in optical density images from ATPase-stained muscle sections. After delineation, fibres may be classified into different histological types (1, 2A, 2B and 2C) using the measurement of their mean optical density (mOD). The densitometric measurements were obtained from three serial histological slides stained under different conditions. The delineation procedure is performed on one of the images: the resulting mask is fitted to the other images using a linear coordinate transform. Along with densitometric measurements, the lesser diameter of the fibres is computed. Both in processing and in analysis, extensive use was made of mathematical morphology tools. All software was implemented on a VICOM digital image processor, extended with a VISIOMORPH morphoprocessor board.

Adenosine Triphosphatases↗
PubMed1989

Transcriptional pattern of 21-hydroxylase gene (P-450C21) during embryonic development, before, and after birth in mice as determined by in situ hybridization.

21-Hydroxylase is a member of the P-450 superfamily of genes involved in the biosynthesis of cortisol and aldosterone in the adrenal cortex. Congenital adrenal hyperplasia, a well-characterized disease, originates from a lack of this enzyme. We present in this report an in situ hybridization study aimed at detecting 21-hydroxylase activity during murine development, from mid gestation to adulthood. Our results demonstrate that even during the embryonic period the adrenal cortex is the only major site of transcription of this enzyme, which is detectable beginning at embryonic day 14. In addition, a peculiar topographical pattern of transcriptional activity, characteristic of the stage of differentiation of the gland, could be drawn. Using a computer-assisted method, we were able to quantitate the relative transcription level at each stage of development. A steady increase in the level of transcription was demonstrated throughout embryonic life to birth, with a drop during the prepubertal period and a final rise at adult age. The possible physiological significance of our findings is discussed.

Adrenal Cortex↗
PubMed1989

Fully automated measurements by light microscopy of tissue sections using a cellular array computer.

Software was developed for the acquisition, segmentation and analysis of microscopic OD-images on a VICOM digital image processor, extended with a VISIOMORPH morphoprocessor board. The delineation algorithms for peroxisomes, lysosomes, and nuclei in liver, kidney, and adrenal gland sections start by thresholding the difference between the original image and a low pass filtered version. The resulting binary mask is then processed by morphological operations in order to produce an object overlay. The efficiency of the programs is evaluated by comparing delineated objects at different OD-levels, created by varying the stain or by multiplying the original pixel values with constant factors. Manual delineation on some images is also used as a reference. More complex algorithms are used for the delineation of muscle fibres in ATP-ase-stained sections and immunocytochemically labelled cells in monolayer preparations. Muscle images from parallel sections with different stainings are matched with a coordinate transform, enabling the transfer of the object mask from a single delineated image to the unprocessed images and thus obtain all necessary information for fibre classification. After segmentation, the OD-images and their object overlays are fed into a data extraction program, measuring for each delineated object user-selected features. Data are sent to a VAX for statistical interpretation.

Adrenal Cortex↗
PubMed1988

Autoradiographic localization of D1 and D2 dopamine receptors in the human brain.

The distribution of dopamine D1 and D2 receptors in several human brain regions was investigated using autoradiography with the radioligands [3H]SCH 23390 and [3H]spiroperidol. The highest densities of both dopamine receptor types are seen in the nucleus caudatus, putamen and nucleus accumbens. Whereas the density of the D2 receptors is similar in the two segments of the globus pallidus, the pars medialis of the globus pallidus contains a three-fold higher concentration of D1 receptors than the pars lateralis. D1 and D2 receptors are present in the amygdala and substantia nigra. Both receptor types are absent in the cerebellum. The thalamus contains low densities of D1 receptors but no D2 receptors. Only D2 receptors are seen in the anterior lobe of the pituitary gland. The whole cerebral cortex is rich in D1 receptors, while D2 receptors, in low concentrations, are confined to the entorhinal area and cingulate cortex.

Aged↗
PubMed1988

Immobilization of acetylcoenzyme A synthetase and the preparation of an enzyme reactor for the synthesis of [11C]acetylcoenzyme A.

The enzyme acetylcoenzyme A synthetase (acetate-CoA ligase (AMP forming), EC 6.2.1.1) from Saccharomyces cerevisiae (baker's yeast) is used for the synthesis of 1 mumol [11C]acetylcoenzyme A. (CoA-[11C]Ac). A screening of the immobilization of the enzyme on differently derivatized controlled pore glass beads (50 nm pore size and 125-180 micron particle size) was performed. Several silanes, spacer arms and terminal reactive groups were tested. The immobilized enzyme was subjected to storage stability tests. From these experiments, the method of choice was selected: immobilization on CNBr-activated controlled pore glass. The immobilized parameters were optimized further to improve the activity of the enzyme-loaded glass beads. The latter were packed in a glass column. The kinetic properties of the column were investigated and optimized to obtain an almost complete conversion of 1 mumol acetate into acetylcoenzyme A (CoA-Ac) within a few minutes. This is realized with an enzyme reactor (13.0 x 0.5 cm) containing 6.12 U active acetylcoenzyme A synthetase immobilized onto 1 g controlled pore glass.

Acetate-CoA Ligase↗
PubMed1988

Prolactin cell subpopulations separated on discontinuous Percoll gradient: an immunocytochemical, biochemical, and physiological characterization.

A single-step procedure was devised to separate PRL cells from the rat anterior pituitary gland. After dissociation, cells were centrifuged on a Percoll gradient. Three layers were recovered. The composition of the different layers was evaluated using immunocytochemistry (with antisera to the six pituitary hormones), and in situ hybridization [with DNA complementary to PRL or to GH messenger RNA (mRNA)]. Both methods yielded identical values. PRL cells were recovered in the lower density layer (layer 1) with a good yield (that is 81% of the total PRL cells of the initial cell suspension) and in addition, markedly enriched (indeed 85% of the cells in layer 1 stained for PRL). A second layer (layer 2: intermediate density) contained most of the remaining PRL cells which were, however, heavily contaminated mainly by GH cells and cells that did not stain for any of the known pituitary hormones. A third layer (layer 3: higher density) was enriched in GH cells to 93% (representing, however, only 10% of the initial pituitary GH cells). In addition, PRL and GH were measured by RIA in culture medium and in cell lysates. Hormone biosynthesis was monitored by polyacrylamide gel electrophoresis and autoradiography after culture in the presence of [35S]methionine. These experiments confirmed that layer 1 was enriched in cells containing, and producing, PRL and depleted from GH cells. Cells in layer 2 contained and produced more GH than PRL. PRL cells from layer 1 responded to dopamine and to vasoactive intestinal polypeptide in the same way as PRL cells in the unseparated pituitary cell population. In contrast PRL cells in layer 2 had a lower basal secretion rate but a higher response to vasoactive intestinal polypeptide. Unless this represents a paracrine effect of non-PRL cells, PRL cells in layer 2 exhibit different properties and may therefore form a distinct subpopulation of PRL cells.

Animals↗
PubMed1988

Automated measurements on cell nuclei in tissue sections. Validation procedures.

A methodology was developed for fully automated measurements of nuclear features in Feulgen-stained tissue sections by means of videomicroscopy and image analysis. Segmentation is performed within one minute on 512 X 512 optical density (OD) images covering about 75 nuclei, resulting in a graphic contour overlay. The corresponding image subset is scanned by an object data extraction program, producing the raw figures for statistical interpretation. The segmentation software was evaluated by three tests, involving comparison with manual delineation and assessment of the influence of OD. Two case studies (ACTH-stimulated adrenal cortex and pancreatic carcinoma) illustrate the biologic accuracy and medical significance of the described methodology.

Adenocarcinoma↗
PubMed1988

Automated measurement of kidney lysosomes by light microscopy. Influence of cyclosporine treatment.

After staining for acid phosphatase, video images were acquired from 1-micron sections of the kidneys of rats treated with cyclosporine. Automated segmentation of the lysosomes was followed by measurements of their area, number and optical density; tubular area was delineated manually. In addition to small lysosomes, representing more than 80% of the total number, all kidneys contained a second population of larger organelles. Cyclosporine treatment with 50 mg/kg/day for one week and with 25 mg/kg/day for three weeks, but not with 25 mg/kg/day for one week, induced a significant increase of lysosomal area, especially in the larger organelles (+200%). In a few animals, this was accompanied by a reduction in the number of lysosomes, suggesting a fusion of the organelles. Other parameters were not consistently modified by treatment. Cyclosporine treatment also elicited the appearance of fat droplets; however, lysosomes were not enlarged in tubular cells containing fat droplets.

Animals↗
PubMed1986

Direct assay for phosphotransacetylase and acetyl-coenzyme A carboxylase by high-performance liquid chromatography.

A simple and specific assay to measure the activity of two coenzyme A derivative-processing enzymes, i.e., phosphotransacetylase (EC 2.3.1.8) and acetyl-coenzyme A carboxylase (EC 6.4.1.2), is described. The assay is based on the HPLC analysis of the short-chain coenzyme A derivatives formed by the enzymatic reaction, viz., acetyl-CoA and malonyl-CoA. For this purpose, ion-pair reversed-phase HPLC conditions are optimized. Furthermore, the influence of several variables on the enzyme reaction is studied in order to get maximum activity. Due to its short analysis time, good selectivity, and chromatogram information, HPLC proves to be an excellent method for the assay of these enzymes.

Acetyl Coenzyme A↗
PubMed1986

Microscale synthesis of nitrogen-13-labeled cisplatin.

A microscale synthesis of [13N]cisplatin (cis-dichlorodiammineplatinum(II), cis-DDP) from cyclotron-produced [13N]ammonia is presented. Temperature, reaction time, ratios, and concentration of reactants have been optimized for each step of the synthesis. Purification is performed by ion exchange chromatography. Radiochemical purity and optimization processes are controlled by high performance liquid chromatography and high performance thin layer chromatography--22 mCi [13N]cisplatin in 10 ml of solution is produced. The entire procedure takes approximately 15 min and the specific activity is approximately 300 mCi/mumole at EOB.

Ammonia↗