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Biomedical subjects

M Fernández de Castro

Publications and source records attributed to M Fernández de Castro.

At least 19 recordsLinked to original sources

Serum level of vascular endothelial growth factor is influenced by erythropoietin treatment in peritoneal dialysis patients. (Grupo de Estudios Peritoneales de Madrid).

Some patients on long-term peritoneal dialysis (PD) develop a hyperpermeability state, owing to peritoneal neoangiogenesis. Vascular endothelial growth factor (VEGF), a potent mitogen for endothelial cells, has been implicated in most diseases characterized by microvascular neoformation. Erythropoietin (EPO) is able to induce endothelial proliferation in vitro. Our aim was to elucidate whether VEGF serum levels are influenced by EPO treatment, and whether VEGF serum level maintains a relationship with peritoneal transport data. We analyzed serum levels of VEGF in 35 PD patients (18 males, 17 females). Mean age was 58 years, with a mean time on PD of 98 +/- 75 months. Of the 35 patients, 19 were on automated peritoneal dialysis, and 16 were on continuous ambulatory peritoneal dialysis. Seven patients had diabetes. Peritoneal transport parameters were: urea mass transfer coefficient (MTC), 19.5 +/- 6.6 mL/min; creatinine MTC, 9.9 +/- 4.7 mL/min; net ultrafiltration, 491 +/- 166 mL per 4-hour dwell. Twenty seven patients were under therapy with recombinant human erythropoietin (rHuEPO). Mean serum VEGF levels were 347 +/- 203 pg/mL (range 66-857 pg/mL), with most patients in the normal range (60-700 pg/mL). VEGF levels did not correlate with age, sex, primary renal disease, diabetes, type of PD, time on PD, peritonitis, and cumulative glucose load. We found no correlation with urea MTC, creatinine MTC, ultrafiltration rate, or protein effluent levels. However, a significant negative correlation with residual renal function was seen (r = -0.39, p < 0.05). Patients treated with rHuEPO showed significantly higher serum levels of VEGF than non treated patients (375 +/- 220 pg/mL vs 251 +/- 75 pg/mL, p < 0.05), although they had similar residual renal function. We conclude that increased serum VEGF levels are associated with EPO treatment. Consequently, VEGF might have a role in the EPO effects found in PD patients. Whether both agents are related to peritoneal neoangiogenesis requires further research.

Biological Transport↗

Ex vivo proliferation of mesothelial cells directly obtained from peritoneal effluent: its relationship with peritoneal antecedents and functional parameters.

The peritoneal membrane requires anatomico-functional integrity to guarantee long-term stability for peritoneal dialysis (PD). Since mesothelial cells (MC) are active cells and the first part of the membrane to contact the dialysate, they are important in maintaining this stability. Mesothelial cells released daily into peritoneal effluent are able to grow in culture. This growth capacity may be related to some of the anatomicofunctional characteristics of each peritoneum. Our aim was to culture mesothelial cells taken from peritoneal effluents drained by 32 PD-stable patients, and relate this growth capacity to individual peritoneal data. Cells were taken from a residual fluid after sedimentation, washed twice with phosphate-buffered saline (PBS), and seeded into 25-cm2 tissue-culture flasks. These flasks were incubated in a humidified 5%-CO2 atmosphere. After MC confluence, cells were detached by trypsinization, passaged into 24-well plates, and finally counted. Cells were identified by morphology and immuno-histochemical characteristics. Cells from 28 out of 32 patients showed an appropriate growth in culture. Mesothelial cell confluence was reached in a mean of 18.2 +/- 8 days. After 7 days of seeding in plate wells, the cell growth showed a significant and progressive increase until day 16. Mesothelial cell growth rate was inversely related to PD duration. Neither peritonitis incidence nor other demographic characteristic were related to MC growth. Creatinine and urea mass transfer coefficients (MTC), but not ultrafiltration (UF) capacity, were significantly related to MC growth rate. In conclusion, the growth in culture of MC taken directly from PD bags is certainly possible. This growth is influenced by some of the intrinsic peritoneal characteristics derived from the peritoneal dialysis process. This tool could be useful in evaluating individual peritoneal conditions and, probably, as a method for peritoneal viability follow-up, although further research is required.

Ascitic Fluid↗

Immunomodulation of peritoneal macrophages by granulocyte-macrophage colony-stimulating factor in humans.

Colony-stimulating factors are growth factors which induce differentiation of the hematopoietic stem cells. Granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulates proliferation and improves functions of neutrophils and monocyte/macrophages. A macrophage submesothelial stratum has been suggested to constitute the first line of peritoneal defense. We have tested whether intraperitoneally administered GM-CSF could increase the number and activation of peritoneal macrophages in peritoneal dialysis patients. Eight stable patients injected 17 micrograms of GM-CSF in each of their four daily CAPD bags over three days. The clinical status, the peritoneal effluent and peripheral blood cell count, membrane receptor expression, phagocytosis activity and cytokine levels were monitored at days 0, 1, 3, 10 and 28. GM-CSF administration caused a large increase in peritoneal macrophage number (89-fold mean increase after 72 hr), returning to baseline seven days after withdrawal. GM-CSF triggered an increase in the expression of CD11b/CD18 (CR3) and its counterreceptor CD54, indicating the cellular progression into a more activated state. Both the number of phagocytic cells (55 +/- 15% to 83 +/- 10%, P < 0.05) and the phagocytic index (137 +/- 29 to 255 +/- 61, P < 0.01) were also augmented. Peritoneal effluent cytokine-chemokine levels demonstrated an increase in IL-6 and MCP-1 levels while TNF-alpha, IL-1, IL-8, MIP-1 alpha and RANTES were not significantly altered. GM-CSF administration did not affect the peritoneal transport of water or solutes. Minor side-effects were registered in two patients. In conclusion, intraperitoneal GM-CSF causes a marked and transient recruitment of primed macrophages into the peritoneum without inducing inflammatory parameters. GM-CSF should improve the peritoneal defensive capacity through potentiation of the effector functions of resident and newly-recruited macrophages.

Adult↗

Presence of different mitogenic activities in the nocturnal peritoneal effluent of patients treated with CAPD.

Peritoneal cells in continuous ambulatory peritoneal dialysis (CAPD) patients are continually being regenerated. We previously reported that nocturnal peritoneal effluent is mitogenic on human and mouse fibroblasts in culture, especially when a comitogen is present. The nature, origin, and role of this mitogenic activity remain undetermined. The data resulting from the addition of different comitogen to dialysate suggest that the peritoneal effluent contains different growth factors with molecular weight greater than 10,000 dalton. Also, the presence of a growth inhibitor is plausible. In conclusion, different growth-promoting and inhibiting activities are present in the peritoneal effluent, suggesting a complex cellular relationship as a result of peritoneal dialysis with unknown consequences.

Animals↗

[Evaluation of the Coulter STKS hematologic autoanalyzer].

PURPOSE: To express the results of an evaluation of the Coulter STKS blood cell counter. MATERIAL AND METHODS: The following data were collected: inaccuracy, carry-over, linearity, comparison with other systems, suspect morphology alarms, stability of calibration, time-stability of samples and working velocity. The protocol recommended by the ICSH was followed for the first four of the above. RESULTS: Good results were achieved for cell counts and derived parameters. The leucocyte differential count showed fair results for neutrophils, lymphocytes, monocytes and eosinophils; false basophilia was usually associated with the presence of abnormal lymphocytes. No differences were found between adult and paediatric samples. The usefulness of the suspect morphology alarms was also evaluated; they were found fairly sensitive for the presence of stabs, large amounts of atypical lymphocytes and blast cells, and moderately sensitive for erythroblasts and platelet clumps. The predictive value of alarms was found high in any type of pathology, but scarcely valuable when considering the pathology indicated by the alarm, such predictive value being noticeably increased upon associated alarms. CONCLUSION: The Coulter STKS cell counter appears as adequate for the haematology laboratory running daily high number of samples with high pathologic rates.

Blood Cell Count↗

[The association of myasthenia gravis and hairy-cell leukemia].

Myasthenia gravis (MG) and tricholeucemia (TL) produce an important immune alteration favoring the appearance of neoplastic and autoimmune disorders. The case of a 53 years old patient who developed type B TL 5 years after the diagnosis of MG is reported. Upon revision of the literature, 37 cases of association of MG and malignant hemopathies, fundamentally lymphoproliferative were found. In general, MG precedes the appearance of the neoplastic process. The autoimmunity and immunodeficiency characteristic of MG probably constitute the pathogenic mechanism leading to the appearance of the neoplastic process.

Bone Marrow↗

[Evaluation of the Sysmex NE-8000 hematologic autoanalyzer].

The results of an evaluation of the Sysmex NE-8000 cell counter are reported. Good results were attained for all values of red and white cell counts. Platelet values were good as well, although samples with thrombocytosis were overstated to some extent. Fair results were found for the white-cell differential count concerning neutrophils, eosinophils and lymphocytes, but false monocytosis were frequently seen in paediatric blood samples, whereas some monocytosis were undetected. The alarms for suspected pathology had only moderate sensitivity when used as the only criterion. The predictive value of such alarms was low, except for the "platelet clumps?" sign.

Automation↗

[Evaluation of a totally automated alternative system for determining the rate of erythrocyte sedimentation].

The Ves-MaticR system (VS) for evaluating erythrocyte sedimentation rate (ESR) is compared to Westergren method. A good correlation between both methods is obtained in spite of small differences observed; the accuracy of the VS is also studied. Concerning the time variation, a small reduction of the ESR in the first seven hours is observed, being significant at 24 hours. The most important advantages of the VS are the near complete disappearance of the contamination risk, the reduction of the time employed in setting up, sedimentation and reading, and the reliability.

Blood Sedimentation↗