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See detailSimultaneous Determination of Deoxyribonucleoside in the Presence of Ribonucleoside Triphosphates in Human Carcinoma Cells by High-Performance Liquid Chromatography
Decoster, L-A; Chen, X; Lejeune, F et al

in Analytical Biochemistry (1999), 270

Simultaneous determination of ribonucleoside and deoxyribonucleoside triphosphates in cells by HPLC is an analytical challenge since the concentration of dNTP present in mammalian cells is several orders ... [more ▼]

Simultaneous determination of ribonucleoside and deoxyribonucleoside triphosphates in cells by HPLC is an analytical challenge since the concentration of dNTP present in mammalian cells is several orders of magnitude lower than the corresponding NTP. Hence, the quantitation of dNTP in cells is generally performed after selective oxidation or removal of the major NTP. The procedures reported so far are lengthy and cumbersome and do not enable the simultaneous determination of NTP. We report the development of a simple, direct HPLC method for the simultaneous determination of dNTP and NTP in colon carcinoma WiDr cell extracts using a stepwise gradient elution ion-pairingHPLCwith uv detection at 260 nm and with a minimal chemical manipulation of cells. Exponentially growing WiDr cells were harvested by centrifugation, rinsed with phosphate- buffered saline, and carefully counted. The pellets were suspended in a known volume of ice-cold water and deproteinized with an equal volume of 6% trichloroacetic acid. The acid cell extracts (corresponding to 2.53 106 cells/100 ml) were centrifuged at 13,000g for 10 min at 4°C. The resulting supernatants were stored at 280°C prior to analysis. Aliquots (100 ml) were neutralized with 4.3 ml saturated Na2CO3 solution prior the injection of 40 ml onto the HPLC column (injection speed 250 ml/min). Chromatographic separations were performed using two Symmetry C18 3.5-mm (2 3 3.9 3 150 mm) columns (Waters), connected in series equipped with a Sentry guard column (3.9 3 20 mm i.d.) filled with the same packing material. The HPLC columns were kept at 30°C. The mobile phase was delivered at a flow rate of 0.5 ml/min, with the following stepwise gradient elution program: % solvent A/solvent B, 100/0 at 0 min 3 100/0 at 1 min 3 36/64 at 5 min 3 31/69 at 90 min 3 31/69 at 105 min 3 0/100 at 106 min 3 0/100 at 120 min; 50/50 MeOH/solvent B from 121 to 130 min; 100% solvent A from 131 to 160 min. Solvent A contained 0.01 M KH2PO4, 0.01 M tetrabutylammonium chloride, and 0.25% MeOH and was adjusted to pH 7.0 (550 ml 10 N NaOH for 1 liter solvent A). Solvent B consisted of 0.1 MKH2PO4, 0.028Mtetrabutylammonium chloride, and 30% MeOH and was neutralized to pH 7.0 (1.4 ml 10 N NaOH for 1 liter solvent B). Even though dNTPs are minor components of cell extracts, satisfactory regression coefficients were obtained for their calibration curves (r2 > 0.99) established with the addition–calibration methods up to 120 pmol/40-ml injection. The applicability of the method was demonstrated by in vitro studies of the modulation of NTP and dNTP pools in WiDr colon carcinoma cell lines exposed to various pharmacological concentrations of cytostatic drugs (i.e., FMdC, IUdR, gemcitabine). In conclusion, this optimized, simplified, analytical method enables the simultaneous quantitation of NTP and dNTP and may represent a valuable tool for the detection of minute alterations of cellular dNTP/NTP pools induced by anticancer/ antiviral drugs and diseases. © 1999 Academic Press [less ▲]

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See detailTwo new aminopeptidases from Ochrobactrum anthropi active on D-alanyl-p-nitroanilide.
Fanuel, L; Thamm, Iris ULg; Kostanjevecki, V et al

in Cellular and Molecular Life Sciences : CMLS (1999), 55(5), 812-8

Two new enzymes which hydrolyse D-alanyl-p-nitroanilide have been detected in Ochrobactrum anthropi LMG7991 extracts. The first enzyme, DmpB, was purified to homogeneity and found to be homologous to the ... [more ▼]

Two new enzymes which hydrolyse D-alanyl-p-nitroanilide have been detected in Ochrobactrum anthropi LMG7991 extracts. The first enzyme, DmpB, was purified to homogeneity and found to be homologous to the Dap protein produced by O. anthropi SCRC C1-38 (ATCC49237). The second enzyme, DmpA, exhibits a similar substrate profile when tested on p-nitroanilide derivatives of glycine and L/D-alanine, but the amounts produced by the Ochrobactrum strain were not sufficient to allow complete purification. Interestingly, the DmpA preparation also exhibited an L-aminopeptidase activity on the tripeptide L-Ala-Gly-Gly but it was not possible to be certain that the same protein was responsible for both p-nitroanilide and peptide hydrolysing activities. The gene encoding the DmpA protein was cloned and sequenced. The deduced protein sequence exhibits varying degrees of similarity with those corresponding to several open reading frames found in the genomes of other prokaryotic organisms, including Mycobacteria. None of these gene products has been isolated or characterised, but a tentative relationship can be proposed with the NylC amidase from Flavobacterium sp. K172. [less ▲]

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See detailExploration isocinétique : analyse des paramètres chiffrés
Croisier, Jean-Louis ULg; Crielaard, Jean-Michel ULg

in Annales de Réadaptation et de Médecine Physique (1999), 42

Introduction : Dans le domaine de l'évaluation de la fonction musculaire, l'isocinétisme permet la quantification de paramètres définissant la qualité de force. Méthode-Objectif : Il s'agit cependant d ... [more ▼]

Introduction : Dans le domaine de l'évaluation de la fonction musculaire, l'isocinétisme permet la quantification de paramètres définissant la qualité de force. Méthode-Objectif : Il s'agit cependant d'optimaliser l'interprétation du bilan isocinétique. Le choix des paramètres les plus représentatifs exige une étude de reproductibilité. Résultats : Lors d'efforts concentriques et excentriques des muscles fléchisseurs et extenseurs du genou, le moment de force maximum répond aux critères qualitatifs souhaités (coefficients de variation faibles); les paramètres de puissance et de travail ne semblent pas apporter d'information complémentaire discriminante en pratique courante. Conclusion: Les auteurs proposent des valeurs de référence et fixent des "limites" précises permettant de distinguer le normal du pathologique, particulièrement pour les paramètres différence bilatérale et rapport agonistes/antagonistes. [less ▲]

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See detailTwist number statistics as an additional measure of habitat perimeter irregularity.
Bogaert, Jan ULg; Van Hecke, P; Moermans, R et al

in Environmental and Ecological Statistics (1999), 6(3), 275-290

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See detailL'ostéoarthrose chez le chien - Pathophysiologie et diagnostic
Loneux, P.; Balligand, Marc ULg

in Annales de Médecine Vétérinaire (1999), (143), 153-160

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See detailEtude du role de Clostridium perfringens dans l'entérotoxémie bovine
Manteca, Christophe; Daube, Georges ULg; Mainil, Jacques ULg

in Bulletin et Mémoires de l'Académie Royale de Médecine de Belgique (1999), 154(6, Pt 2),

Bovine enterotoxaemia is an acute to peracute syndrome occurring mainly in calves and characterized by the sudden or very rapid death of the calf, with colics, convulsions and nervous disorders as ... [more ▼]

Bovine enterotoxaemia is an acute to peracute syndrome occurring mainly in calves and characterized by the sudden or very rapid death of the calf, with colics, convulsions and nervous disorders as clinical signs, if any. The most pronounced lesion is a necrohaemorrhagic enteritis of the jejunum, the ileum, and sometimes the colon. Suckling beef calves are the most frequently affected ones. In 67% of the 78 field cases investigated, some kind of stress was observed 24 to 36 hours prior to the death: change in diet or pasture, vaccination... The most frequently isolated bacteria, and the one isolated in highest numbers, was non-sporulated non-enterotoxigenic toxinotype A Clostridium perfringens. Reproduction of the lesions was successful in a ligated intestinal loop assay in one calf with a few of these strains, more especially with one of them, which was shown later to produce another recently described toxin, the beta 2 toxin. A role for this beta 2 toxin in bovine enterotoxaemia is thus speculated for future research. [less ▲]

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See detailNonresponsive Generalized Bacterial Infection Associated with Systemic Lupus Erythematosus in a Beauceron
Clercx, Cécile ULg; Mc Entee, Kathleen ULg; Gilbert, S. et al

in Journal of the American Animal Hospital Association (1999), 35(3, May-Jun), 220222224-8

A case of concurrent canine systemic lupus erythematosus (SLE) and generalized bacterial infection in a six-year-old female Beauceron is reported. The dog presented with purulent nasal and ocular ... [more ▼]

A case of concurrent canine systemic lupus erythematosus (SLE) and generalized bacterial infection in a six-year-old female Beauceron is reported. The dog presented with purulent nasal and ocular discharges, skin lesions (including seborrhea, hyperkeratotic areas, and papules as well as ecchymoses around the eyes, on both sides of the pinnae, and on the vulva), generalized lymph node enlargement, a mitral murmur, and lameness. Later, facial swelling, a retrobulbar abscess, and a cough also developed. Occurrence of a generalized bacterial infection was established by culture of group-C, beta-hemolytic Streptococcus from the throat, the mouth, a biopsy site (popliteal lymph node area), the retrobulbar abscess, and the lung. The diagnosis of SLE was based on the clinical signs and particularly on the occurrence of antinuclear antibody (ANA) and antidoublestranded-desoxyribonucleic acid (ds-DNA) antibody. Interestingly, the latter type of antibodies were also detected in two young female puppies whelped by this dog. Salient histological findings included an extreme cell depletion of the lymph nodes and spleen and severe pneumonitis and peribronchiolitis. The results of this case indicate that a definite diagnosis of canine SLE can, at times, be made on the basis of the presence of serum ANA and ds-DNA antibodies. [less ▲]

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See detailL'ostéoarthrose chez le chien - Traitement
Loneux, P.; Balligand, Marc ULg

in Annales de Médecine Vétérinaire (1999), (143), 163-175

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See detailRadiothérapie externe ou anti-inflammatoire non-stéroïdien pour la prévention des ossifications hétérotopiques après prothèse totale de hanche?
Zouhair, A; Ozsahin, M; Mouhsine, E et al

in Schweizerische Medizinische Wochenschrift (1999), 129(9), 370-376

External radiotherapy or non-steroid antiinflammatory drugs for prevention of heterotopic ossification following total hip replacement Heterotopic ossification (HO) is defined as the development of ... [more ▼]

External radiotherapy or non-steroid antiinflammatory drugs for prevention of heterotopic ossification following total hip replacement Heterotopic ossification (HO) is defined as the development of abnormal ossification in soft tissues. HO is a common disease after total hip replacement. Many therapeutic modalities have been proposed to prevent HO. The most commonly used modalities are nonsteroidal anti-inflammatory drugs (NSAID) or ionizing radiation administered just before or immediately after total hip replacement. As far as external radiation therapy is concerned, there are several published randomized studies aimed at investigating its efficacy and timing related to surgery, and at comparing ionizing irradiation to NSAID. In this article we review the published data in order to define guidelines which could be used in daily practice for the choice of prophylactic treatment against HO. [less ▲]

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See detailLes troubles cognitifs consécutifs à un trauma crânien léger chez l'enfant
Closset, Annette; Van der Linden, Martial ULg; Misson, Jean-Paul ULg

in Percentile (1999), 4

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See detailSingle motor unit H reflexes recorded in thenar muscles at rest.
Wang, François-Charles ULg; Delwaide, Paul ULg

in Muscle & nerve (1999), 22(2), 291-2

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See detailEffectiveness of an acute pain service inception in a general hospital
Bardiau, Françoise ULg; Braeckman, M.M.; SEIDEL, Laurence ULg et al

in Journal of Clinical Anesthesia (1999), 11(7), 583-589

STUDY OBJECTIVES: To assess the effects of an Acute Pain Service (APS) inception on postoperative pain management in a general teaching hospital using pain indicators as performance measures. DESIGN: Open ... [more ▼]

STUDY OBJECTIVES: To assess the effects of an Acute Pain Service (APS) inception on postoperative pain management in a general teaching hospital using pain indicators as performance measures. DESIGN: Open, prospective, nonrandomized, observational study. SETTING: Postanesthesia Care Unit, surgical wards of University Hospital Center of Charleroi. PATIENTS: 1304 patients in the pre-APS inception phase and 671 patients after its implemention who have undergone various types of surgery (orthopedics, gynecology, urology, neurosurgery, stomatology, ear, nose, and throat, ophthalmic, abdominal, vascular-thoracic, plastic, and maxillofacial). INTERVENTIONS: An APS, nurse-based, anesthesiologist-supervised model was devised, based on the concept that postoperative pain relief can be greatly improved by providing in-service training for surgical nursing staff and optimal use of systemic analgesics. MEASUREMENTS AND MAIN RESULTS: Postoperative pain was assessed using a visual analog scale (VAS) every 4 hours for 72 hours in the two phases. Analgesic consumption was registered at the same time. Time-related VAS scores were summarized using several pain indicators. There was an overall improvement in the pain scores after APS inception. The differences were most pronounced, around 50%, in patients undergoing vascular, maxillofacial, gynecologic, and urologic surgeries, and stomatology. Regular administration of paracetamol and nonsteroidal antiinflammatory drugs decreased morphine consumption in the second phase. CONCLUSION: This study validates the benefits of a formal APS, using continuous monitoring of rest pain intensity and analgesic consumption in the postoperative period. Results not only support previous research findings but also offer outcome-based tools to evaluate current practices as compared with desired outcomes. [less ▲]

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See detailMorphométrie du pied du cheval et maladie naviculaire
Collin, B.; Gabriel, Annick ULg

in Bulletin de l'Académie Vétérinaire de France (1999), 72

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