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Biomedical subjects

J Comas

Publications and source records attributed to J Comas.

At least 19 recordsLinked to original sources

A hybrid supervisory system to support WWTP operation: implementation and validation.

Integrated operation of Wastewater Treatment Plants is still far from being solved. A reasonable proposal should link advanced and robust control algorithms to some knowledge-based techniques, allocating the detailed engineering to numerical computations, while delegating the logical analysis and reasoning to supervisory intelligent systems. This paper describes the development and implementation of a knowledge-based Hybrid Supervisory System to support the operation of a real Wastewater Treatment Plant. The system integrates different reasoning modules, overcoming the limitations in the use of each single technique, while providing an agent based architecture with additional modularity and independence. It is structured into three separated levels: data gathering, diagnosis, and decision support. The different tasks of the system are performed in a seven-step cycle: data gathering and update, diagnosis, supervision, prediction, communication, actuation, and evaluation phase. In spite of certain reservations of the scientific community about the use of these techniques, the system is successfully performing real-time support to the operation of the Granollers facility since September 1999. Results of the first four-month validation period are shown and discussed. An example of the system behavior is also shown in the paper. The conclusions indicate the key steps which are necessary to transfer the system to another facility.

Algorithms↗

Rapid flow cytometry--Nile red assessment of PHA cellular content and heterogeneity in cultures of Pseudomonas aeruginosa 47T2 (NCIB 40044) grown in waste frying oil.

The accumulation of cytoplasmic polyhydroxyalkanoates (PHAs) and the heterogeneity ofbacterial populations were analysed by flow cytometry and SYTO-13 and Nile red staining in rhamnolipid-producing Pseudomonas aeruginosa cultures grown in waste frying oil as carbon source. A combination of SYTO-13 and Nile red fluorescence with cytometric forward and side scatter values may allow increases in the final production of polyhydroxyalkanoates (PHA) by two basic mechanisms: (i) rapid assessment of polyhydroxyalkanoate content and (ii) definition of flow cytometric cell sorting protocols to select high polyhydroxyalkanoate (PHA)-producing strains. We report a rapid (less than 30 min) flow cytometric assessment of PHAs in Pseudomonas aeruginosa 47T2 following Nile red staining: (i) to estimate cellular PHAs content; (ii) to study heterogeneity of the batch cultures producing PHAs and (iii) to establish the basis for sorting sub-populations with a high capacity to accumulate PHAs.

Cooking↗

Development of a case-based system for the supervision of an activated sludge process.

The development of a case-based reasoning system for the supervision of an activated sludge process is presented here. The methodology proposed permits the use of past experiences to solve new problems that arise in the process. These experiences are classified as cases or situations. The adaptation of cases and the generation of new cases are used to tune the response of the system and to learn from the new information generated by the process. The case and the case library definition, the initial seed, the search and retrieval process, the adaptation, the action, the evaluation and the learning steps are presented and outlined. The process studied is the wastewater treatment plant of Girona, Spain. Two examples of the response of the system to two different operational situations are presented. The paper also outlines the integration of different fields in a multidisciplinary approach as the most optimal solution to ensure the successful control and supervision of complex processes like the activated sludge process. With this aim the integration of an array of specific supervisory intelligent systems (for the logical analysis and reasoning) and numerical computations for detailed engineering is suggested.

Bacteria↗

Second-order functions are the simplest correlations between flow cytometric light scatter and bacterial diameter.

Second-order mathematical relationships between bacterial cell diameter determined by electric particle analyser and flow cytometric forward light scatter in axenic cultures are obtained and discussed. Since it is technically impossible today to obtain both measurements for each individual cell, standard regression techniques cannot be applied. To overcome this limitation, we assume that these two parameters are related by a monotone increasing function that enables their mathematical relationships to be studied. Our conclusion is that forward light scatter data cannot be linearly transformed into bacterial size values by an accurate and universal function. However, second-order relationships seem to be the simplest satisfactory relationships between cell diameter and forward light scatter in eubacteria.

Bacteria↗

Flow cytometric analysis of a marine LAS-degrading consortia.

The specific nucleic acid fluorochrome SYTO-13 was used in flow cytometric analysis to assess changes in the density and heterogeneity of marine bacterial populations which biodegrade linear alkylbenzene sulphonate (LAS). Seawater samples with LAS and incubated in the laboratory (20 degrees C, 100 rpm, 30 days) were used to monitor LAS-degrading consortia. Flow cytometric studies and culture methods were used to characterize the LAS degrading bacterioplankton consortia. Fluorescence and scatter signals enabled us to define three regions (R1, R2 and R3) in the dual parameter cytograms. The distribution of the bacterial counts in these regions allowed us to monitor the formation and evolution of the consortia.

Alkanesulfonic Acids↗

Evaluation of free radical production, mitochondrial membrane potential and cytoplasmic calcium in mammalian neurons by flow cytometry.

The overexcitation of glutamate receptors is believed to be the cause of several neurodegenerative disorders. The determination of calcium fluxes, mitochondrial membrane potential (MMP) variations or the production of reactive oxygen species (ROS) in mammalian cells are usually measured during the development of potentially useful drugs that might interfere in the events induced by glutamate receptor activation. By using flow cytometry with dissociated cerebellar granule cells, we have developed a rapid and economical method to measure changes in biochemical parameters that are involved in neuronal cell death. The formation of intracellular ROS is measured using 2',7'-dichlorofluorescin diacetate (DCFH-DA). The mitochondrial membrane potential is assessed by the retention of rhodamine 123 (Rh123), a specific fluorescent cationic dye that is readily sequestered by active mitochondria, depending on their transmembrane potential. Finally, intracellular calcium increases are detected by using the calcium-selective indicator Indo-1. Cell viability is also assessed by using propidium iodide (PI) which stains DNA strands of permeabilized cells. This method might be useful for the screening of new drugs with potential neuroprotective activity, with improved cost/effectiveness ratio compared to other techniques.

Animals↗

Assessment of the effects of gramicidin, formaldehyde, and surfactants on Escherichia coli by flow cytometry using nucleic acid and membrane potential dyes.

Two membrane potential sensitive dyes (Rhodamine 123 and bis-oxonol) and three nucleic acid dyes (propidium iodide, SYTO-13, and SYTO-17) were used to assess the effect of surfactants on Escherichia coli. The ability of E. coli to be stained by these probes was validated at different physiological states. Propidium iodide was used to assess the integrity of cell envelopes. Two double staining methods based on propidium iodide with SYTO-13 and bis-oxonol with SYTO-17 were used to improve the discrimination between bacteria and micelles or aggregated particles generated by the presence of surfactants. A rapid (1 h contact time between cells and surfactants, and less than 5 min for staining and obtaining data) Rhodamine 123 flow cytometric assay was developed to assess the bactericidal effect of surfactants.

Anti-Infective Agents, Local↗

Mitochondrial membrane potential measurement in rat cerebellar neurons by flow cytometry.

Mitochondrial membrane potential (MMP) in dissociated rat cerebellar neurons was measured using rhodamine 123 (Rh 123) as fluorescent dye, and flow cytometry. Dye distribution was studied by confocal scanning microscopy. Propidium iodide (PI)-marked cells (dead cells) were not stained by Rh 123, while the green fluorescence of living cells was restricted to mitochondria. Incubation of cells with different ionophores resulted in a maximal inhibition of Rh 123 fluorescence of 27.0 +/- 5.9% (valinomycin), 55.6 +/- 7.2% (ionomycin), and 37.3 +/- 5.1% (gramicidin). Ionophores decreased cell viability at high concentrations, measured as the number of propidium iodide-marked cells. Exposure of cell suspensions to the mitochondrial specific uncoupling agent CCCP caused a decrease in Rh 123 fluorescence (40 +/- 6.1%). Conversely, oxidative stress induced by H2O2 did not affect Rh 123 fluorescence. Impairment of glucose bioavailability reduced Rh 123 fluorescence. 2-Deoxy-D-glucose decreased the MMP with a maximal inhibition of 24.0 +/- 4.4%. Lack of glucose in the incubation medium also resulted in a decrease in MMP. Moreover, application of L-glutamate and N-methyl-D-aspartate (NMDA) (the excitatory amino acids) decreased Rh 123 uptake in a dose-dependent manner, which suggests that the measurement of MMP in dissociated cerebellar neurons by flow cytometry is a suitable method to detect the activity of drugs acting on glutamate receptors.

Animals↗

Comparison of four lymphocyte isolation methods applied to rodent T cell subpopulations and B cells.

The aim of this study was to establish the validity of four lymphocyte isolation methods. The effects of three different erythrocyte lysing methods commonly used in the analysis of human cells, namely, lysis by ammonium chloride (AC), Becton Dickinson lysis (BDL) and the Coulter Q-Prep (CQP) preparation system were established by flow cytometry on rat lymphocyte subsets. The results were compared with those obtained with a Ficoll-Isopaque (FI) density gradient procedure adapted for use with rat cells. Lymphocyte isolation by AC or FI gradient was performed before labelling the lymphocyte subpopulations, whereas the BDL and CQP methods were performed after staining the cells in whole blood. The FI gradient yielded the lowest CD5+, CD4+ and CD25+ cell percentages. On the other hand AC lysis produced higher percentages of T cells and lower percentages of B cells than the other methods studied. The percentages obtained after BDL or CQP methods for T lymphocyte subsets and B cells were found to be reproducible. The commercial methods (BDL and CQP) are faster but rather expensive, whereas AC lysis and FI gradient separations are cheap and particularly useful when there is a requirement to culture the cells.

Animals↗

Surgery for congenital mitral valve disease in the first year of life.

Between 1980 and 1993, 20 patients less than 1 year of age underwent operations for congenital mitral valve disease. Ten patients had congenital mitral incompetence and 10 had congenital mitral stenosis. Mean age was 6.6 +/- 3.4 months and mean weight was 5.6 +/- 1.5 kg. Atrioventricular canal defects, univentricular heart, class III/IV hypoplastic left heart syndrome, discordant atrioventricular and ventriculoarterial connections, and acquired mitral valve disease were excluded. Indications for operation were intractable heart failure or severe pulmonary hypertension, or both. Associated lesions, present in 90% of the patients, had been corrected by a previous operation in seven. In congenital mitral incompetence there was normal leaflet motion (n = 3), leaflet prolapse (n = 2), and restricted leaflet motion (n = 5). In congenital mitral stenosis anatomic abnormalities were parachute mitral valve (n = 4), typical mitral stenosis (n = 3), hammock mitral valve (n = 2), and supramitral ring (n = 1). Mitral valve repair was initially performed in 19 patients and valve replacement in one with hammock valve. Concurrent repair of associated lesions was performed in 12 patients. The operative mortality rate was zero. There were six early reoperations in five patients for mitral valve replacement (n = 4), a second repair (n = 1), and prosthetic valve thrombectomy (n = 1). One late death occurred 9 months after valve replacement. Late reoperations for mitral valve replacement (n = 2), aortic valve replacement (n = 1), mitral valve repair (n = 2), subaortic stenosis resection (n = 1), and second mitral valve replacement (n = 1) were performed in five patients. Actuarial freedom from reoperation is 58.0% +/- 11.3% (70% confidence limits 46.9% to 68.9%) at 7 years. After a mean follow-up of 67.6 +/- 42.8 months, 94% of living patients are in New York Heart Association class I. Doppler echocardiographic studies among the 13 patients with a native mitral valve show mitral incompetence of greater than moderate degree in one patient and no significant residual mitral stenosis. Overall, six patients have mitral prosthetic valves with a mean transprosthetic gradient of 6.2 +/- 3.7 mm Hg. These results show that surgical treatment for congenital mitral valve disease in the first year of life can be performed with low mortality. Valve repair is a realistic goal in about 70% of patients and possibly more with increased experience. Reoperation rate is still high and is related to complexity of mitral lesions and associated anomalies, but late functional results are encouraging.

Follow-Up Studies↗

Flow cytometric assessment of Escherichia coli and Salmonella typhimurium starvation-survival in seawater using rhodamine 123, propidium iodide, and oxonol.

The use of flow cytometry in microbiology allows rapid characterization of cells from a nonhomogeneous population. A method based on flow cytometry to assess the effects of lethal agents and the bacterial survival in starved cultures through the use of membrane potential-sensitive dyes and a nucleic acid marker is presented. The use of propidium iodide, rhodamine, and oxonol has facilitated the differentiation of cells of Escherichia coli and Salmonella typhimurium of various states of vitality following various treatments (heat, sonication, electroporation, and incubation with gramicidin) and during starvation in artificial seawater. The fluorescence intensity is directly correlated with viable cell counts for rhodamine 123 labelling, whereas oxonol and propidium iodide labelling is inversely correlated with viable counts. The distribution of rhodamine and oxonol uptake during starvation-survival clearly indicates that single-species starved bacteria are heterogeneous populations, and flow cytometry can be a fundamental tool for quantifying this heterogeneity.

Escherichia coli↗

Variations in flow cytometric forward scatter signals and cell size in batch cultures of Escherichia coli.

Flow cytometry was used to study the lag, exponential, stationary and death phases of non-fixed cultures of Escherichia coli. Fluctuations in the forward angle scatter signal (FALS) were compared with cell size as measured by scanning electron microscopy at low temperature and image analysis. A correlation between FALS and cell size was not observed, although a correlation (r = -0.8) was obtained between FALS and the age of the culture for the first eleven days of incubation. Marked increases in FALS were observed during the lag phase, which were attributed both to changes in size and changes in structure or chemical composition. The distribution of FALS for all culture phases was asymmetric, and was associated with the cell size distribution.

Cell Size↗

A flow cytometric study of the rat Yoshida AH-130 ascites hepatoma.

The implantation of the Yoshida AH-130 ascites hepatoma to rats results in a marked reduction in body weight in the tumour-bearing hosts. This is associated with an important reduction in both food intake and energetic efficiency in the last period of tumour growth. The growth of the tumour mass has a clear initial exponential phase, in which the maximum cell density is reached, with the majority of cells being in the synthetic phase (S) followed by most of the cells being in the G0/G1 phase, as determined using flow cytometry.

Animals↗

Anatomic correction of transposition of the great arteries in neonates.

OBJECTIVES: This retrospective study attempts to assess the results of the neonatal anatomic repair of transposition of the great arteries by a single institution. BACKGROUND: Anatomic correction of transposition of the great arteries by means of the arterial switch operation is now widely accepted as the therapeutic method of choice. METHODS: Four hundred thirty-two consecutive neonates underwent an arterial switch operation for various forms of transposition of the great arteries. There were 362 neonates with transposition and intact ventricular septum, 47 with a ventricular septal defect, 6 with intact ventricular septum and coarctation of the aorta and 17 with ventricular septal defect and coarctation. Among patients with coarctation, 18 underwent a single-stage repair through median sternotomy. The mean age was 13.1 +/- 4.2 days. Coronary artery distribution was described according to the origin and initial course of the arteries. RESULTS: Overall in-hospital mortality was 7.8% (34 patients) and was 7.6% for transposition with intact ventricular septum, 8.5% for transposition with ventricular septal defect and 13.3% for transposition with ventricular septal defect and coarctation. Univariate analysis of risk factors revealed that coronary anatomy was the main determinant for operative survival. A mean follow-up time of 44 +/- 19 months was achieved in all but five survivors. More than 95% were in New York Heart Association functional class I, without medication and with normal left ventricular function. Reoperation was performed in 20 patients: early reoperation (< 30 days) in 4 and late reoperation in 16. Actuarial survival rates at 5 years were 91.3% (transposition with intact ventricular septum) and 81.06% (transposition with ventricular septal defect). As well, freedom from reoperation at 5 years was 96.8% (transposition with intact ventricular septum) and 84.6% (transposition with ventricular septal defect). CONCLUSIONS: The arterial switch operation is feasible in almost all forms of transposition of the great arteries in neonates as primary and definitive repair. Palliative surgery is recommended in cases with complex intracardiac anatomy not amenable to early repair.

Aorta↗

Comparison of different types of cardioplegia and reperfusion on myocardial metabolism and free radical activity.

Current techniques of myocardial protection during global ischemia include hypothermia, cardioplegic arrest and controlled reperfusion. To compare different types of cardioplegia and reperfusion techniques we measured the levels of adenine nucleotides and malondialdehyde (MDA, as free radical activity) in 33 patients undergoing heart surgery. The patients were randomized in three groups according to the characteristics of cardioplegia and reperfusion: cold blood cardioplegia with unmodified blood reperfusion (control group, 11 patients), crystalloid cardioplegia and reperfusion (Hôpital Lariboisière protocol, 11 patients) and crystalloid cardioplegia with allopurinol enriched blood reperfusion (Hôpital Broussais protocol, 11 patients). Myocardial biopsy specimens were obtained before cardioplegic arrest (preischemic values), at the end of ischemia and after 30 minutes of reperfusion. Biopsy specimens were analyzed by high performance liquid chromatography for levels of adenine nucleotides and MDA. In the three groups, the preischemic values of adenine nucleotides and MDA were not significantly different. For AMP and ADP concentrations neither treatment nor biopsy-time effects appeared. ATP concentration decreased significantly with biopsy-time without specific treatment effect. For MDA concentration neither treatment nor biopsy-time effects were observed. This study suggests that there is no statistically significant difference between any of the three cardioplegia and reperfusion techniques for either ATP or MDA; the three reperfusion techniques limit the free radical activity but do not prevent the fall in high energy phosphates.

Adenine Nucleotides↗

[Value of coronary reperfusion in cases of very low cardiac output following coronary artery bypass grafts].

From January 1986 to December 1988, 244 patients had a coronary artery revascularization at St Joseph Hospital. Four patients (1.8%) died in post operative time (less than 30 days). None died directly from cardiogenic shock. Five patients had a severe low cardiac output (2 h to 48 h post-op) and needed cardiac resuscitation. These patients were immediately taken to the operating room. The coronary bypasses were explored (2 technical errors corrected), the metabolic disorders were treated and a coronary reperfusion was done with the Buckberg's technique. All the patients could be weaned from cardiopulmonary bypass, two with intra aortic balloon pumping. One patient died of digestive complication the 10th postoperative day. The four other patients survived and are in good clinical condition. The severe low cardiac output after coronary revascularization can be reversible with resuscitation in operating room, short assistance, and coronary reperfusion.

Aged↗