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F Vara

Publications and source records attributed to F Vara.

At least 19 recordsLinked to original sources
PubMed2003

[Influence of different pharmacological agents in the ex vivo proliferation of mesothelial cells obtained from the peritoneal effluent of patients treated with peritoneal dialysis].

UNLABELLED: Mesothelial cells (MC) are the first peritoneal membrane barrier in contact with dialysate. The aim of this study was to analyze the in vitro capacity of different pharmacological agents to modify the ex vivo proliferation of MC obtained from the peritoneal effluent of patients treated with peritoneal dialysis (PD). MATERIAL AND METHODS: Thirty cultures of MC taken from nocturnal peritoneal effluent were performed. After identification, MC are subcultured in 24 multi-well plates, adding the different exogenous agents. Proliferative capacity and cell morphology were estimated on day 16th of culture. The agents evaluated were insulin, IGF-1, tamoxifen, labetalol, carvedilol, enalapril and losartan. RESULTS: Insulin shows a dose-dependent effect on MC growth, with a limit that is stimulated by the addition of fetal bovine serum (FBS). Concentrations higher than 100 micrograms/ml, are not associated with further growth, even with cell damage. In contrast, the wide range of IGF-1 dose used did not affect to MC proliferation. Tamoxifen causes negative effects on MC growth just a very high doses, not resembling doses in clinical practice. Labetalol does not modify MC proliferation used under therapeutic calculated range. However, concentrations higher than 40 micrograms/ml showed a negative influence on growth, behaving as lethal doses that over 100 micrograms/ml. The addition of FBS attenuates this effect. These effects were very similar to that caused by carvedilol addition. Enalapril and losartan act as antiproliferative agents for MC. This effect is potentiated with angiotensin II, reaching lethal concentrations increasing the dose. In conclusion, mesothelial cell growth ex vivo taken from nocturnal peritoneal effluent on PD patients is an useful tool to explore the effects of any pharmacological agent on the biology of the cell of the peritoneum. The agents used had any influence in the proliferation capacity of mesothelial cells.

Ascitic Fluid↗
PubMed1999

[A prospective study of meningitis diagnosed in a 3rd-level hospital during a 1-year period].

OBJECTIVE: One-year prospective observational study of meningitis diagnosed at a third level hospital. PATIENTS AND METHODS: All patients with a cerebrospinal fluid (CSF) specimen with cyto-biochemical characteristics and clinical picture consistent with meningitis were included in the study. They were followed from admission to hospital up to discharge or exitus. The epidemiologic characteristics of patients, etiology, related risk factors and predisposing situations, CSF characteristics, clinical manifestations, clinical course, and antibiotic susceptibility of the causative agents were analyzed. RESULTS: Ninety-five cases were included. Seventy-six (69.4%) were community acquired and 29 (30.5%) nosocomially acquired meningitis. Among community acquired meningitis, 31 (46.9%) were of bacterial origin (8 N. meningitidis, 3 H. influenzae, 2 S. pneumoniae, 1 Streptococcus group B, 1 Listeria monocytogenes, 1 Staphylococcus aureus, and 1 Brucella spp.); CSF culture was negative in 14 cases (41.2%). In most cases neither risk factor nor predisposing situations were detected. Patients with purulent meningitis and negative CSF culture had a significantly lower number of complications than patients with positive CSF culture. Among patients previously treated with beta-lactam antibiotics (8 cases) the probability of a negative CSF culture was greater than among not treated patients (OR 16.00, 95% CI 1.45-764.68; p = 0.011). The remaining cyto-biochemical characteristics were similar in both groups. Thirty-five cases (53.03%) of community acquisition were lymphocytic meningitis (31 viral, 3 tuberculous, and 1 luetic meningitis). Among nosocomial cases (29 cases, 30.5%), most were caused by gram-negative bacilli and microorganisms of the Staphylococcus genus. Fourteen cases (48.2%) were related to some type of neurosurgical procedure. Overall, only two exitus cases were recorded. CONCLUSIONS: The etiologic agents of community acquired meningitis are mainly N. meningitidis, S. pneumoniae and Haemophilus influenzae. The previous antibiotic therapy did not influence thy cyto-biochemical characteristics of CSF but it did influence the yielding of culture. Meningitis with negative CSF culture has a significantly lower number of complications. The availability of a Neurosurgery Department at a hospital confers a change in the epidemiologic spectrum of diagnosed meningitis, with a higher incidence of nosocomial meningitis. In our environment, a substantial proportion of cases due to Staphylococcus microorganisms was observed.

Adolescent↗
PubMed1998

Jugular venous oxygen monitoring: a helpful technique in the early diagnosis of a traumatic carotid-cavernous sinus fistula.

This report describes the early diagnosis of a right traumatic carotid-cavernous sinus fistula (CCSF) in a patient with head injury manifested as an acute increase in right jugular venous oxygen saturation and with no ophthalmic clinical signs. High values of jugular venous oxygen saturation must be cautiously interpreted with the clinical examination and computed tomographic findings to establish an accurate diagnosis of hyperemia with or without a CCSF.

Adult↗
PubMed1998

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↗
PubMed1996

[Videothoracoscopy in the surgical treatment of pneumothorax: report of 46 cases].

We report our 3-years experience with video-thoracoscopy for pneumothorax. Forty-six endoscopies (34 men, 12 women; mean age 27 +/- 9 years) were performed. Patients were under general anesthesia and intubation was selective. The conventional approach was used, with 3 incisions through which the camera and endoscopic instruments were inserted. Results were good in 39 (84.7%) patients. Pneumothorax recurred in 4 (8.7%) patients during the 3 years of follow-up. Thoracotomy was performed in 3 patients, because of lack of lung expansion in 1, persistent leak in another and encapsulated empyema in the third. Mean hospital stay after surgery was 7.3 days. Video-thoracoscopy can be considered a good therapeutic alternative to thoracotomy in the surgical treatment of patients with pneumothorax.

Adult↗
PubMed1996

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↗
PubMed1995

The influence of adrenalin on the pharmacokinetics of interpleurally administered lidocaine in patients with pancreatic neoplasia.

The influence of adrenalin on the pharmacokinetics of lidocaine given interpleurally to 10 patients with pancreatic neoplasia was studied. Five patients received an interpleural dose of lidocaine (200 mg; control group), and 5 patients received an interpleural dose of lidocaine (200 mg) plus adrenalin (1:200,000). Plasma and cerebrospinal fluid (CSF) levels of lidocaine were measured before and at specified times (up to 8 hours) after the dose. The analytical technique was radioimmunoassay; and plasma and CSF data were assessed using noncompartmental analysis. The drug was quickly absorbed into the plasma in the control group (Cmax = 2.76 +/- 0.10 microgram/mL at 0.33 +/- 0.14 hours after administration); whereas drug access to CSF was decreased and occurred slowly (Cmax = 0.32 +/- 0.07 microgram/mL at 1.66 +/- 1.35 hours). The drug was eliminated more quickly from plasma than from CSF, with half-lives of 1.71 +/- 0.43 hours and 3.86 +/- 1.27 hours, respectively. The simultaneous administration of adrenalin delayed absorption (tmax = 0.91 +/- 0.52 hours). The drug elimination half-lives in plasma and CSF of this group increased to 3.22 +/- 1.22 hours and 8.71 +/- 3.28 hours, respectively. The duration of the analgesia, evaluated as the time until the patient needed another dose, increased from 8.2 +/- 1.5 hours in the control group to 9.7 +/- 1.3 hours in the group that received adrenalin. From these results the levels that would be reached on a multiple-dose regimen (D = 200 mg, tau = 8 hours) were predicted.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗
PubMed1994

Cell populations present in the nocturnal peritoneal effluent of patients on continuous ambulatory peritoneal dialysis and their relationship with peritoneal function and incidence of peritonitis.

OBJECTIVE: To study the relationship between peritoneal effluent cells and infection rate and to relate this population with functional characteristics. DESIGN: Prospective, longitudinal, and comparative study. SETTING: Outpatient continuous ambulatory peritoneal dialysis (CAPD) unit of a university medical center. PARTICIPANTS: Seventy-one uninfected patients, treated for 0-156 months on CAPD, in stable condition were studied (33 female, 38 male). INTERVENTIONS: Nocturnal peritoneal effluent (NPE) was drained with EDTA (2.5 mmol/L) at 37 degrees C and centrifuged at 2500 rpm for 9 minutes. MEASUREMENTS: Accumulated peritoneal inflammation days/year and ultrafiltration/diffusion (mass transfer coefficients (MTCs) for small molecules) capacities were recorded. Cellular count (cells/night) was performed using a Neubauer chamber. Macrophage function was assessed by cytochemical (lysosomal enzyme content: ANAE, beta-glucuronidase, acid phosphatase) and immunohistochemical procedures (expression of membrane antigens, CD4, 11b, 11c, 14, 16, 25, 35, and 71). RESULTS: The macrophage is the most frequently appearing cell in the NPE. Cell count decreases over time on CAPD (from 20 x 10(6) to 5 x 10(6) after the first year). Intrapatient variability was low, but interpatient differences were marked. Mesothelial cell count remained stable over time (0.25-0.5 x 10(6)). Four of our patients showed a "transforming" change in these cells. Previous incidence of peritonitis and values of functional measurements did not correlate with cell count or expressions of macrophage function (lysosome enzyme content and percentage of cells expressing different membrane antigens). CONCLUSION: There is difficulty interpreting the results on peritoneal effluent cells and their relationship with the incidence of peritonitis and functional characteristics of the peritoneum. No definite conclusions can be drawn other than the great interpatient and intrapatient variability. The presence of abnormal peritoneal cells with undetermined origin and function suggests the need for periodic studies of peritoneal effluent cells on long-term CAPD patients.

Cell Count↗
PubMed1993

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↗
PubMed1991

Effects of the addition to dialysate of calcitriol H-R-erythropoietin (EPO) and interferon-alpha-2b (IFN) on mitogenic activity of peritoneal effluent.

The presence of cellular growth factors in peritoneal effluents from CAPD patients have been described. The purpose of this paper has been to evaluate the effects of mitogenic activity of peritoneal effluents of the in vivo and in vitro additions of Calcitriol, EPO and IFN. These drugs were added to nocturnal peritoneal dialysate (in vivo) and to nocturnal effluent (in vitro) and then these fluids were tested for mitogenic-induced capacity on mice and human fibroblasts in culture. Our data show that the addition of Calcitriol and Erythropoietin have little effects on peritoneal effluent behaviour. Interferon alpha-2b seem to be useful for decreasing human fibroblasts growth.

Animals↗
PubMed1990

Fibroblast growth factor stimulates protein kinase C in quiescent 3T3 cells without Ca2+ mobilization or inositol phosphate accumulation.

To elucidate the transmembrane signalling processes initiated by fibroblast growth factor (FGF), we have studied the effect of recombinant basic FGF (bFGF) on various early events associated with mitogenesis in Swiss 3T3 fibroblasts. bFGF, at mitogenic concentrations, neither induced Ca2+ mobilization from intracellular stores nor increased the accumulation of inositol phosphates. In contrast, bFGF stimulated the phosphorylation of the Mr 80,000 (80K) cellular protein which is a major substrate of protein kinase C. This effect was potentiated by the diacylglycerol kinase inhibitor R59022. Two-dimensional polyacrylamide gel electrophoresis and phosphopeptide mapping showed that the 80K phosphoproteins generated in response to bFGF, bombesin, and phorbol 12,13-dibutyrate were indistinguishable. Down-regulation of protein kinase C prevented bFGF stimulation of 80K phosphorylation. Other protein kinase C-dependent early events such as transmodulation of the epidermal growth factor receptor, cytoplasmic alkalinization, inhibition of vasopressin induced increase in cytosolic [Ca2+], and enhancement of cAMP accumulation in response to forskolin were also induced by bFGF. Similar results were obtained when bFGF was added to quiescent cultures of tertiary mouse embryo fibroblasts. We conclude that bFGF stimulates protein kinase C through a signal transduction pathway distinct from inositol phospholipid turnover and Ca2+ mobilization.

Animals↗
PubMed1989

Insulin influence (used as an additive to dialysate) on the mitogenic-induced effect of the peritoneal effluent in CAPD patients.

Peritoneal effluent in CAPD patients is the result of the interaction between dialysate and patient through the peritoneal membrane. Among the factors transferred from the patient to dialysate are solutes capable of stimulating fibroblasts: Interleukin-I, Interferon-gamma, and other proteins. Insulin is a well known mitogenic coadjuvant able to act sinergistically with other mitogens in the stimulation of fibroblast proliferation under experimental conditions. Our objective has been to study the effect of insulin added to dialysate in vivo on the in vitro mitogen-induced capacity of the nocturnal peritoneal effluent in 8 diabetics. Two different samples were studied: basal and with in vivo added insulin with the patients' usual doses. A Swiss 3T3 line of mice fibroblasts was used for studies, adding 50 microliters of peritoneal effluent. To confirm in vivo potential mitogenic activity, we also added PDBu (Phorbol dibuturate), a well known mitogenic agent, to both samples. Furthermore, another growth factor, EGF (Epidermal growth factor), was also added to 5 other samples. Insulin addition to the CAPD bag transforms a non-mitogenic peritoneal effluent into a mitogenic one (mean: 4.4 times over basal). At the doses used in vivo there is no linear correlation between insulin dose and DNA synthesis (r: -0.28, NS). Both PDBu and EGF in vitro addition induced a remarkable increment in the mitogenic capacity of the peritoneal effluent. We conclude that insulin added to CAPD bags behaves as a remarkable mitogenic coadjuvant for mice fibroblasts. Consequently, its long-term and universal clinical use should be reconsidered.

Animals↗
PubMed1987

Inhibition of the proton pumping ATPases of yeast and oat root plasma membranes by dicyclohexylcarbodiimide.

The inhibition of the proton-pumping ATPases of yeast and oat root plasma membranes by dicyclohexylcarbodiimide (DCCD) can be correlated with the covalent incorporation of the inhibitor. Full inhibition of the yeast enzyme required the incorporation of about 1 mol DCCD/mol of the ATPase polypeptide of 100 kDa. A kinetic study of the interaction of DCCD with the yeast and oat ATPases indicates a second-order rate constant of about 500 M-1 min-1 and a stoichiometry of 1 mol DCCD/mol of enzyme, in agreement with the amount of DCCD incorporated by the yeast enzyme. It is proposed that DCCD reacts with a single carboxylic group present in a hydrophobic region of these proton-pumping ATPases and which could participate in proton binding and transport.

Amino Acid Sequence↗
PubMed1985

Stimulation of Na+/H+ antiport activity by epidermal growth factor and insulin occurs without activation of protein kinase C.

Addition of phorbol 12, 13 dibutyrate (PBt2) or a combination of epidermal growth factor (EGF) and insulin to quiescent cultures of Swiss 3T3 cells increased intracellular pH in a Na+-dependent fashion. In contrast to PBt2, EGF plus insulin failed to stimulate protein kinase C and elicited the ionic response in cells lacking this enzyme. We suggest that the stimulation of the Na+/H+ antiport in Swiss 3T3 cells is mediated by at least two separate pathways, only one of which is dependent upon the activation of protein kinase C.

Animals↗
PubMed1985

Ionic responses rapidly elicited by activation of protein kinase C in quiescent Swiss 3T3 cells.

Diacylglycerol and phorbol esters activate protein kinase C in intact cells. We report here that addition of the synthetic diacylglycerol 1-oleoyl-2-acetylglycerol (OAG) to quiescent cultures of Swiss 3T3 cells caused a marked increase in the rate of ouabain-sensitive 86Rb+ uptake, a measure of the activity of the Na+/K+ pump. The effect was dose-dependent and could be detected after 1 min of exposure to the diacylglycerol. OAG stimulated Na+ influx via an amiloride-sensitive pathway and increased intracellular pH by 0.15 pH unit. Phorbol 12,13-dibutyrate (PBt2) also enhanced ouabain-sensitive 86Rb+ uptake and amiloride-sensitive 22Na+ influx. Prolonged treatment (40 hr) of 3T3 cells with PBt2 at a saturating dose, which reduces the number of PBt2 binding sites and protein kinase C activity, abolished the ionic response of the cells to a subsequent addition of either OAG or PBt2. Appropriate controls using acid "loads" and the Na+ ionophore monensin showed that the function of the Na+/H+ antiport system and of the Na+/K+ pump was not impaired in the PBt2-desensitized cells. We suggest that activation of protein kinase C elicits, either directly or indirectly, enhanced Na+/H+ antiport activity, which, in turn, leads to Na+ influx, intracellular pH modulation, and stimulation of the Na+/K+ pump.

Amiloride↗
PubMed1983

Phosphorylated intermediate of the ATPase of plant plasma membranes.

A partially purified preparation of the plant plasma membrane ATPase was phosphorylated when incubated with [gamma-32P]ATP. The phosphoprotein formed has the characteristics of an enzyme intermediate because of its rapidity of phosphorylation and dephosphorylation. The sensitivity of the phosphoenzyme bond to alkaline pH and to hydroxylamine indicates that it is an acylphosphate. Both the ATPase activity and the phosphorylation of the enzyme exhibited an apparent Km value of 0.3 mM ATP. When the phosphorylated enzyme was analyzed by electrophoresis in sodium dodecyl sulfate, only one major band with a molecular weight of about 105,000 contained radioactivity. These results indicate that the plant plasma membrane ATPase has a subunit composition and reaction mechanism similar to the cation-pumping ATPases of animal and fungal plasma membranes.

Adenosine Triphosphatases↗
PubMed1982

Partial purification and properties of the proton-translocating ATPase of plant plasma membranes.

The plasma membrane ATPase of plant cells has been postulated to operate as an electrogenic proton pump which derives the co-transport of nutrients with protons and which possibly catalyzes K+ transport (Poole, R. J. (1978) Annu. Rev. Plant Physiol. 29, 437-460). In addition, the enzyme seems to determine cell growth after hormonal stimulation by acidifying the external medium (Marré, E. (1979) Annu. Rev. Plant Physiol. 30, 273-288). In order to substantiate this important physiological role, the ATPase from oat root plasma membranes has been solubilized with a zwitterionic detergent and partially purified. A polypeptide of 93,000 daltons was enriched in the course of the purification. The enzyme was completely specific for ATP as substrate and it was inhibited by vanadate, diethylstilbestrol, and dicyclohexylcarbodiimide but not by oligomycin or ouabain. The ATPase activity was stimulated by K+ but this occurred only at acidic pH and the effect was less than 100%. After reconstitution of proteoliposomes by a freeze-thaw-sonication procedure, proton transport driven by ATP was demonstrated by the quenching of acridine dye fluorescence. Proton transport occurred in the absence of K+ and its electrogenic nature was demonstrated by the requirement for permeant ions (nitrate or K+ with valinomycin). It is suggested that the enzyme is an electrogenic proton pump, somewhat stimulated by K+, but not involved in the transport of this cation.

Adenosine Triphosphatases↗