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W Goossens

Publications and source records attributed to W Goossens.

At least 37 records · Page 2Linked to original sources

The prevention of erythrocyte swelling upon dilution after freezing and thawing.

Cellular swelling of erythrocytes exposed to Me2SO during freezing and thawing may lead to hemolysis upon dilution of the cryoprotectant with pure electrolyte buffer. Excessive cell swelling is effectively avoided by exposing the RBC to the nonpenetrating sorbitol after thawing and before dilution. Due to the initial reduction in volume by sorbitol, cell swelling upon dilution may not cause hemolysis particularly with concentrations of 0.05 to 0.15 M of sorbitol in the diluting electrolyte buffer. Membrane damage incurred during freezing and thawing is particularly pronounced with the older red cell population, while the younger population membrane integrity can be preserved to an optimal degree.

Blood Preservation↗

Ultrastructural alterations in the aqueous outflow pathway of adult buphthalmic rabbits.

The aqueous outflow pathway of adult rabbit eyes with congenital glaucoma (buphthalmos) was examined by light microscopy and by scanning and transmission electron microscopy. The morphology of the buphthalmic rabbit aqueous outflow pathway was markedly abnormal when examined at 6 months, 1 yr, and 2 yr displaying apparent loss and/or compression of the iris pillars, dilation of the intertrabecular spaces, loss of endothelial cell-to-cell association and disorganization of trabecular lamellae, and posterior displacement of the aqueous plexus. In addition, the trabecular meshwork lamellae were observed only adjacent to the sclera and the inner portion of the trabecular meshwork was limited to swirls of collagen with scattered cells. These morphological findings suggest that the disease process in the rabbit principally involves an alteration in the differentiation and maintenance of the structural integrity of the trabecular meshwork. The loss of structural support of the buphthalmic trabecular meshwork may be a factor in the wide variation in intraocular pressure and may allow for compression of the trabecular meshwork against the aqueous plexus.

Animals↗

K2- or K3-EDTA: the anticoagulant of choice in routine haematology?

The choice of K2- or K3-EDTA as the preferred anticoagulant for blood count remains controversial. We compared the effect of different concentrations of both anticoagulants on normal blood. In optimal conditions (appropriate anticoagulant concentration and measurements done between 1 and 4 h after phlebotomy), no marked differences are seen between either EDTA salt. Important discrepancies appear, however, in less optimal conditions, as often happens in day to day practice. The packed cell volume, when measured on centrifuged blood, decreases with increasing anticoagulant concentrations and this is most pronounced with the K3 salt. This phenomenon has been reported by different authors and is ascribed to shrinking of erythrocytes in an hypertonic medium. Automated instruments react in a different way, their MCV is not influenced by K3-EDTA concentrations up to ten times normal, while K2-EDTA, at high concentrations, results in a slight increase in MCV, as measured with three of the instruments. With most instruments, the accuracy of the white cell count is not markedly influenced. However, when measured with the Unipath CD 3000 all tested blood samples, taken in high K3 (but not in K2) concentrations, showed an appreciable decrease in the leucocyte count (to less than 50% of the original value, at a concentration of 15 g/l, 24 h after blood collection). Measurement of RDW and automated differentials is also influenced by the choice of anticoagulant when determinations are done in less than optimal conditions. We conclude that the choice of anticoagulant, its use at an appropriate concentration and the age of the blood sample are important matters and should be given due consideration.(ABSTRACT TRUNCATED AT 250 WORDS)

Anticoagulants↗

Monocyte counting: discrepancies in results obtained with different automated instruments.

To determine the accuracy of several methods for measuring the monocyte count, the results obtained by a number of different automated cell counters were analysed. Considerable discrepancies occurred for monocyte counts obtained in normal blood among the counters. The results of a visual monocyte count on a total of 800 leucocytes were used as the reference method. The technique of measuring the monocyte count by using dual staining with monoclonal antibodies CD45 and CD14 provided the closest agreement with the reference method. Six other automated counting systems were assessed. Two of these systems (Coulter VCS and Technicon H1) gave results, which, although under-estimating monocytosis, correlated well with the results obtained by the reference technique. A third system (Toa Sysmex NE-8000) gave unreliable results. Three of the automated systems evaluated measured a "third population"--that is, monocytes together with other leucocytes. One of these systems (Ortho ELT 1500), overestimated the count, as expected, but correlated well with the reference method. The second of these "third population counters" (Coulter S Plus IV) correlated moderately well with the reference monocytosis, while the Toa Sysmex E-5000 correlated poorly. It is clear that problems exist in the evaluation of different instruments for counting monocytes. An accurate and reliable reference method is a pre-requisite to evaluate this aspect of cell counters. As the visual method is too cumbersome a different reference method would be useful. Based on the results of this study, it is suggested that the technique using fluorescence labelled monoclonal antibodies should be regarded as an acceptable alternative.

Adult↗

[Vitamin B12 (cobalamin)-deficiency in the elderly].

In order to determine the relevance of low vitamin B12 concentration in elderly persons (greater than 65 years), serum vitamin B12 levels were measured in 292 consecutive patients upon their admission to a geriatric ward. Eighty (27%) of them were anaemic (haemoglobin less than or equal to 7.14 mmol/l). The vitamin B12 concentration was not statistically different between males and females, nor between the anaemic and non-anaemic groups. Twenty-two patients (7.5%) had a low vitamin B12 level; of these, 10 had a haemoglobin concentration higher than 7.14 mmol/l and a mean corpuscular volume lower than 98 fl. In addition, serum vitamin B12 levels were also measured in 178 consecutive anaemic patients hospitalized in the same ward. Seventeen (9.5%) of them had a vitamin B12 deficiency although in 11 the MCV was lower than 98 fl. Hypersegmented polymorphonuclear leukocytes were rarely observed in the peripheral blood smear. Pernicious anaemia could only be diagnosed in 1 patient. Serum vitamin B12 estimation may be necessary to detect vitamin B12 deficiency but a low level requires additional diagnostic investigation.

Aged↗

Iodine and selenium deficiency associated with cretinism in northern Zaire.

Selenium status was determined in an endemic-goiter area and in a control area of Zaire. Compared with the reference values of a noniodine-deficient area, serum selenium in subjects living in the core of the northern Zaire endemic-goiter belt (Karawa villages) was seven times lower in 52 school-children and similarly low in 23 cretins; erythrocyte glutathione peroxidase (RBC-GPX) was five times lower in schoolchildren and still two times lower in cretins (P = 0.004). In a less severely iodine-deficient city of the same endemia (Businga), selenium status was moderately altered. RBC-GPX activity was linearly associated with serum selenium concentration up to a value of 1140 nmol/L and leveled off at approximately 15 U/g Hb at greater selenium concentration. At Karawa villages, selenium supplementation normalized both the serum selenium and the RBC-GPX. This combined iodine and selenium deficiency could be associated with the elevated frequency of endemic myxedematous cretinism in Central Africa.

Adolescent↗

Reticulocyte count using thiazole orange. A flow cytometry method.

Recently flow cytometry techniques have been developed to replace the microscope reticulocyte count. We used thiazole orange, a RNA binding fluorochrome, to discriminate reticulocytes from mature erythrocytes. Thiazole orange and the Retic-COUNT software package were evaluated for performance of routine analysis on different flow instruments. The applied methodology analysed 10(4) cells semi-automatically in an easily performed manner. Consistent results were obtained with dipotassium EDTA anticoagulated blood (stable for 30 h after venesection), with incubation times in thiazole orange solution ranging from 2 to 7 h at 25 degrees C. This allowed flexibility in specimen collection and storage and assay performance with no change in results. Changes of incubation temperature up to 30 degrees C had no measurable effect. The values obtained showed good linearity, precision and accuracy for normal, low and high reticulocyte counts. However interferences were observed: RBC autofluorescence, nucleated RBC, Howell-Jolly bodies, high leucocyte count, high platelet count and giant platelets, all falsely increased the number of reticulocytes. These artifacts were eliminated by software gate corrections, thus leaving less than 5% of the specimen to be reanalysed by the microscopic method. The thiazole orange flow cytometric method was determined to be a fast, reliable method for the routine clinical quantitation of reticulocytes.

Benzothiazoles↗

Plasma atrial natriuretic peptide and the renin-aldosterone system during long-term administration of the diuretic xipamide in man.

We have studied the effect of xipamide on plasma alpha-atrial natriuretic peptide and the renin-aldosterone-kallikrein system in twelve healthy men, using a double-blind cross-over design. After a run-in period on placebo for 1 week the subjects were treated with either placebo (n = 6) or xipamide 20 mg once daily (n = 6) for 16 weeks and were then switched to the alternative medication for another 16 weeks. The plasma concentration of alpha-atrial natriuretic peptide fell after 1 week of xipamide administration and increased during prolonged xipamide administration but remained suppressed. The changes in plasma alpha-ANP observed after 1 week of xipamide were negatively correlated with the changes in haematocrit and haemoglobin. Plasma renin activity (PRA), aldosterone concentration (PAC), and urinary excretion of aldosterone and kallikrein increased after 1 week of xipamide administration, levelled off during the second and fourth weeks, but remained elevated during further prolonged xipamide administration for 16 weeks. The xipamide-induced changes in PRA and PAC were positively correlated with the changes in the haematocrit and haemoglobin. Our data suggest that the changes in plasma renin, aldosterone, and alpha-atrial natriuretic peptide during xipamide administration may be related to diuretic-induced volume contraction.

Aldosterone↗

Erythrocyte 2,3-diphosphoglycerate concentration before and after a marathon in men.

Erythrocyte 2,3-diphosphoglycerate (2,3-DPG) concentration was studied in 23 runners before and after a marathon race. Blood samples were drawn from an antecubital vein the morning before the race (baseline), at 3 p.m. 2 h before the start, on finishing, and 12 and 36 h later. Compared to the baseline values, erythrocyte 2,3-DPG concentration was increased (p less than 0.001) immediately after the marathon from 4.62 +/- 0.14 to 5.56 +/- 0.13 mumol.ml-1 RBC and remained elevated 12 h later (5.45 +/- 0.14 mumol.ml-1 RBC): it returned to prerace values 36 h after completion of the marathon.

2,3-Diphosphoglycerate↗

Effects of training on erythrocyte 2,3-diphosphoglycerate in normal men.

The erythrocyte 2,3-diphosphoglycerate concentration (2,3-DPG) and the activity of red cell hexokinase, pyruvate kinase, glucose-6 phosphate dehydrogenase and glutathione reductase were studied in 27 normal volunteers before and after 2 and 4 months of physical endurance training. The 4 months of training increased maximal oxygen uptake and physical working capacity (PWC130) by 16% (p less than 0.001) and 29% (p less than 0.001) respectively. Resting heart rate was decreased (p less than 0.001) by 11 beats.min-1. With 2 months of training the erythrocyte 2,3-DPG concentration increased by 9% (p less than 0.001); with 4 months training the increase was only 4% (p less than 0.05). The training-induced increase in red cell 2,3-DPG was not accompanied by enhanced activity of erythrocyte hexokinase, pyruvate kinase, glucose-6 phosphate dehydrogenase or glutathione reductase. It is concluded that the rise in red cell 2,3-DPG induced by physical endurance training is not due to activation of red cell glycolytic enzymes or the enzymes involved in the pentose-phosphate cycle.

2,3-Diphosphoglycerate↗

Indicators of cell breakdown in plasma of men during and after a marathon race.

Plasma indicators of muscle cell leakage and of hemolysis were studied in 23 runners before and after a marathon race. Blood samples were drawn from an antecubital vein the morning before the race (baseline), at 3 p.m., i.e., 2 h before the start, on arrival, 12 and 36 h, and 7 days later. Compared with the baseline values, the plasma creatinine phosphokinase MM and MB subfractions, aldolase and glutamicoxaloacetic transaminase activity were increased immediately after the race, rose further 12 h after the marathon, and remained elevated the race, rose further 12 h after the marathon, and remained elevated 36 h and 7 days later. The plasma lactate dehydrogenase activity and myoglobin concentration were increased on arrival and returned to the pre-race activity 7 days after the marathon. Compared with the pre-race values, the plasma haptoglobin concentration was decreased immediately and 12 h after the marathon. Our data show that indicators of muscle cell leakage and of hemolysis in plasma, withdrawn after a marathon race, remained elevated for up to 7 days after the race.

Adult↗

Erythrocyte and leucocyte sodium and potassium transport systems during long-term diuretic administration in men.

The effect of xipamide on the intracellular concentration and transmembrane fluxes of Na+ and K+ was studied in 12 normal male subjects, using a double-blind cross-over design. After a run-in period on placebo for 1 week, the subjects were treated with either placebo (n = 6) or xipamide 20 mg once a day (n = 6) for 16 weeks and were then switched to the alternative medication for another 16 weeks. The intra-erythrocyte and intra-leucocyte Na+ concentration was increased by 11 and 7%, respectively, during xipamide administration, while the intracellular K+ concentration was decreased by 3 and 4%, respectively. No significant effect of xipamide could however be demonstrated on the ouabain-sensitive, bumetanide-sensitive or ouabain-bumetanide-resistant 86Rb uptake and on the maximal 3H-ouabain binding in erythrocytes and leucocytes. The red cell Na+-Li+ countertransport was also not changed in the xipamide-treated subjects. Our data suggest that the increased intracellular Na+ concentration and the decreased K+ concentration in red and white blood cells of xipamide-treated subjects cannot be attributed to changes in the activity of the Na+ pump, the Na+-K+ cotransport or Na+-Li+ countertransport system or to changes in the number of active Na+ pump units.

Adult↗

The effect of blood transfusion on cerebral blood-flow in preterm infants: a Doppler study.

Stable infants with anaemia needing a transfusion with adult red blood-cells were studied to elucidate changes in brain blood-flow velocity. Within 24 hours and at five to six days following transfusion a substantial mean flow velocity reduction was observed. Haemodynamic factors contributing to the reduction were an increase in cerebrovascular resistance and an increase in whole blood viscosity, as reflected by a raised pulsatility index. Transfusion with adult red blood-cells causes an elevation in haemoglobin concentration, thereby increasing the total oxygen-carrying capacity of arterial blood; however, this lowers the concentration of fetal haemoglobin which possesses a higher affinity for oxygen. Since cerebral oxygen transport is equal to the product of cerebral blood-flow and arterial oxygen content, this finding suggests the existence of a homeostatic mechanism for cerebral oxygen transport. The actual amount of cerebral oxygen transport was found to increase progressively as the percentage of fetal haemoglobin rose above 30 per cent. At higher fetal haemoglobin levels, appropriate elevations in cerebral blood-flow occurred, causing an increase in the supply of oxygen to the brain.

Blood Flow Velocity↗

Effects of physical endurance training on the plasma renin-angiotensin-aldosterone system in normal man.

The effect of physical endurance training on the plasma renin-angiotensin-aldosterone system was studied in 27 normal sedentary volunteers aged between 20 and 55 years, using a randomized two-period cross-over study design. After 4 months of training (2.5 h/week), peak oxygen uptake and physical working capacity at a heart rate of 130 beats/min were increased by 16% (P less than 0.01) and 29% (P less than 0.001) respectively, whereas resting heart rate was decreased by 15% (P less than 0.001). The plasma noradrenaline concentration and haematocrit were both decreased (P less than 0.01) after training. For the total group of subjects, the small decreases in plasma renin activity (PRA) and in the plasma concentrations of angiotensin-I, angiotensin-II and aldosterone were not statistically significant. However, the change in PRA during the training period was negatively correlated with the increase in physical working capacity (r = -0.49, P less than 0.01), suggesting that PRA decreased only in those subjects with the greatest increase in exercise capacity. Also, the change in plasma aldosterone during training was negatively related to the rise in physical working capacity (r = -0.57, P less than 0.01). Furthermore, the changes in plasma angiotensin-I (r = 0.75), angiotensin-II (r = 0.49) and aldosterone (r = 0.43) during the training period correlated positively with the change in PRA. It is concluded that physical endurance training, leading to a substantial gain of physical working capacity, suppresses the plasma renin-angiotensin-aldosterone system in normal man.

Adult↗

Dietary sodium variation, erythrocyte cationic transport and plasma renin-aldosterone in men.

Erythrocyte concentrations and fluxes of sodium and potassium were investigated in normal white male subjects during dietary sodium restriction and repletion, each period lasting for 16 weeks. Intraerythrocyte sodium concentration decreased and red cell ouabain-sensitive 86Rubidium-uptake increased during dietary sodium restriction while no significant changes were observed in the total, furosemide-resistant and furosemide-sensitive sodium-efflux and the sodium, lithium-countertransport. The decrease in intraerythrocyte sodium concentration could have resulted from the observed increase in sodium, potassium-ATPase pump activity. The latter increase could have been secondary to the early decrease in a digitalis-like plasma inhibitor and the later increase could have been facilitated by the late rise in the intracellular adenosine triphosphate concentration, which is the energy supplier for this pump. During the subsequent first month of sodium repletion intraerythrocyte sodium concentration remained low. Red cell ouabain-sensitive 86Rubidium-uptake and adenosine triphosphate concentration remained elevated and returned to baseline only after 16 weeks. This long-term effect suggests either the involvement of a mechanism which can only be slowly reversible or a mechanism which is irreversible so that normalization takes place only when new red cells are released into the circulation.

Adult↗

Fluorescence-labeled lectins, glycoconjugates, and the development of the mouse AOP.

The development of the aqueous outflow pathway (AOP) in early postnatal mouse eyes was examined for the presence of a variety of lectin receptors using fluorescein isothiocyanate (FITC) conjugated lectins, 1 micron araldite plastic sections, and computer-aided fluorescence photography. The trabecular meshwork anlage (days 1-4) was characterized by the presence of loosely arranged cells and an extracellular matrix that exhibited intense areas of Con A- and RCA-lectin staining, and absence of WGA- and LPA-lectin staining. By day 6, trabecular meshwork LPA- and WGA-positive materials were observed as focal areas of staining. By day 10, LPA- and WGA-positive materials were present as diffuse areas of staining, as the AOP differentiated into an organized and functional biological filter. The age-dependent pattern of LPA- and WGA-positive materials indicated that there were time-dependent points in the synthesis of glycoconjugates in the developing AOP. The results suggest: The composition and/or conformation of the glycoconjugates on cells and extracellular matrix changed as the AOP differentiated into a functional tissue. The use of FITC lectins as biological markers for studies of the AOP provided information on the potential role of glycoconjugates in the development of the normal AOP. Modification in the type, amount, and distribution of glycoconjugates may provide a basis for understanding the cellular mechanisms of abnormal development of the AOP, eg, congenital glaucoma.

Animals↗

Sodium and potassium fluxes and concentrations in erythrocytes of normal subjects during prolonged sodium depletion and repletion.

The erythrocyte concentration and fluxes of sodium and potassium were investigated in normal white male subjects during dietary sodium restriction and repletion, each period lasting for 16 weeks. During dietary sodium restriction the intra-erythrocyte sodium concentration decreased and the red cell ouabain-sensitive 86Rb-uptake increased; no significant changes were observed in the ouabain-insensitive fluxes of sodium such as the total, frusemide-resistant and frusemide-sensitive Na+-efflux and the Na+, Li+-countertransport. The decrease in the intra-erythrocyte sodium concentration could result from an increased Na+,K+-ATPase pump activity. The latter increase could be secondary to the early decrease in a digitalis-like plasma inhibitor and the late increase could be facilitated by the late rise in the intracellular ATP concentration, which is the energy supplier for this pump. During the subsequent first month of sodium repletion the intra-erythrocyte sodium concentration remained low, the red cell ouabain-sensitive 86Rb-uptake and ATP concentration remained elevated and returned to baseline only after 16 weeks. This long-term effect suggests either the involvement of a mechanism which can only be slowly reversible or a mechanism which is irreversible so that normalization takes place only when new red cells are released into the circulation.

Adult↗