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Biomedical subjects

J A Fernandez

Publications and source records attributed to J A Fernandez.

At least 19 recordsLinked to original sources
PubMed2004

Digestibility of carbohydrates in growing pigs: a comparison between the T-cannula and the steered ileo-caecal valve cannula.

We compared the determination of ileal and total tract digestibility of carbohydrates in five experimental diets using a double 5 x 5 Latin square design involving a total of 10 cannulated pigs; half of the pigs were equipped with a simple T-cannula and the other half with steered ileo-caecal valve (SICV)-cannula. The diets consisted of nitrogen-free mixture and soya bean meal, sunflower meal, peas or rape seed cake diluted to about 180 g/kg DM protein with the nitrogen-free mixture. There was no significant difference in the digestibility values using the two types of cannulas with regard to organic matter, sugars (sum of glucose, fructose and sucrose), alpha-galactosides (sum of raffinose, stachyose and verbascose), starch, cellulose, total non-cellulosic polysaccharides (NCP) and insoluble NCP constituents. The digestibility values for the NCP residues arabinose and galactose, however, were estimated higher but with a lower variability with the SICV-cannula compared with the T-cannula. The type of cannula did no influence the estimation of the total tract digestibility for any of the major dietary constituents, but the total tract digestibility was slightly more variable when the pigs were equipped with the SICV-cannula compared with the T-cannula. There was no difference in the ileal digestibility of sugars, alpha-galactosides, cellulose and the NCP arabinose and uronic acids residues among the experimental diets, while the ileal digestibility of starch and the remaining NCP sugar residues varied between diets. The total tract digestibility was complete for sugars, alpha-galactosides and starch, whereas the digestibility of the cell wall constituents varied in accordance with the polymeric composition of the cell walls. It was concluded that ileal digesta samples from SICV-cannula are more homogenous than those from the T-cannula. In cases where the precision of each determination is crucial, the SICV-cannula should be the option.

Animal Feed↗
PubMed2002

Uptake and elimination of radiocaesium in fish and the "size effect.".

A number of hypotheses have previously been developed concerning the rates of uptake and elimination of radiocaesium (137Cs) in fish. These include the influence of potassium and other water chemical parameters on both uptake and elimination, and the effect of fish size on accumulation. In order to test these hypotheses, we have assembled a data set comprising more than 1,000 measurements of radiocaesium (137Cs) in predatory fish (perch, pike and brown trout) in nine European lakes during the years after Chernobyl. These data have been analysed using simple models for uptake and excretion of 137Cs in fish, showing that: 1. Fish-water concentration factors (CF) were inversely proportional to potassium [K+] concentration of the different lakes, in agreement with previous studies. 2. The uptake rate of 137Cs in fish was negatively correlated with lake [K+], but excretion rate was independent of [K+]. 3. Lower than expected CF values were found in one lake, Iso Valkjärvi, Finland. This is attributed to inhibition of the K+ (and therefore 137Cs) high affinity transport system in aquatic plants and fish by low pH and/or low Ca2+. 4. The inclusion of fish weight as a parameter in our dynamic model significantly improves the ability of the model to fit the observed measurements of 137Cs. 5. The model developed from the above hypotheses was able to fit the data from nine different lakes to within approximately a factor of 3 of the observed values.

Animals↗
PubMed2001

Anti-inflammatory, antithrombotic, and neuroprotective effects of activated protein C in a murine model of focal ischemic stroke.

BACKGROUND: Activated protein C (APC) contributes to systemic anticoagulant and anti-inflammatory activities. APC may reduce organ damage by inhibiting thrombin generation and leukocyte activation. Neutrophils and cerebrovascular thrombosis contribute to ischemic neuronal injury, suggesting that APC may be a potential protective agent for stroke. METHODS AND RESULTS: We examined the effects of APC in a murine model of focal ischemia. After middle cerebral artery occlusion/reperfusion, the average survival time in controls was 13.6 hours. Animals that received purified human plasma-derived APC 2 mg/kg IV either 15 minutes before or 10 minutes after stroke induction survived 24 hours and were killed for neuropathological analysis. APC 2 mg/kg given before or after onset of ischemia restored cerebral blood flow, reduced brain infarct volume (59% to 69%; P:<0.003) and brain edema (50% to 61%; P:<0.05), eliminated brain infiltration with neutrophils, and reduced the number of fibrin-positive cerebral vessels by 57% (P:<0.05) and 25% (nonsignificant), respectively. The neuroprotective effect of APC was dose-dependent and associated with significant inhibition of ICAM-1 expression on ischemic cerebral blood vessels (eg, 61% inhibition with 2 mg/kg APC). Intracerebral bleeding was not observed with APC. CONCLUSIONS: APC exerts anti-inflammatory, antithrombotic, and neuroprotective effects in stroke. Central effects of APC are likely to be related to improved maintenance of the blood-brain barrier to neutrophils and to reduced microvascular obstructions and fibrin deposition.

Animals↗
PubMed2001

A mechanistic sub-model predicting the influence of potassium on radiocesium uptake in aquatic biota.

It is often argued that the quality of science is related to the possibilities of making accurate predictions. It has also long been argued that due to the complex nature of ecosystems, it will never be possible to predict important target variables, especially with more comprehensive dynamic models. New results in radioecology have, however, demonstrated that those arguments are no longer valid. The key to the predictive success lies in the structuring of the model. The accident at Chernobyl has, in fact, provided science with an intriguing opportunity to study how the pulse of 137Cs is transported through ecosystem pathways, thus revealing the basic structure of these ecosystems, i.e. which are the key and the less-important pathways. It is paradoxical to conclude that the Chernobyl accident is, perhaps, the most important factor behind the revolution in predictive ecosystem modelling which lies behind the decrease in the uncertainty factor from 10 to 0.25-0.5. The sub-model for the potassium moderator presented in this paper is an example of a mechanistically based sub-model used within the framework of a more comprehensive lake model for 137Cs. The K-moderator presented is derived from the existing knowledge on ion transport in biological membranes and takes into account ion equilibria modelled by the Nernst equation and the uptake kinetics quantified by the Michaelis-Menten model. It provides the type of structure to this overall lake model that helps to explain the excellent predictive power of this model.

Animals↗
PubMed2001

Susceptibility testing of pathogenic fungi with itraconazole: a process analysis of test variables.

A 2(10-5) fractional factorial model was used to investigate the influence of 10 process variables in broth microdilution susceptibility tests with itraconazole against eight isolates of Candida species and six isolates of filamentous fungi in two growth media. An analysis of variance (ANOVA) indicated that glucose concentration and incubation time both significantly influenced control turbidity optical density (OD) values for most of the Candida spp. isolates, while incubation in >10% CO(2) versus ambient air, incubation temperature and inoculum size significantly influenced these OD values for about half of the yeast isolates. Control OD values for the mould isolates were most influenced by incubation time and temperature, and by occlusion of the wells with an adhesive sticker. Three statistical approaches, ANOVA, rank transformation and Mann-Whitney U-test, were used to assess the influence of the variable combinations on MIC, determined with a 50% growth reduction end-point. Incubation temperature and time, glucose concentration and inoculum size were the variables that most often affected susceptibility results to the level of statistical significance; however, the supplier of RPMI 1640 medium, the use of adhesive stickers and the atmosphere of incubation significantly influenced the MIC for some isolates. The medium used to prepare the test inoculum, the solvent used to prepare the stock solution and the shape of the microdilution plate wells significantly affected outcome, but only sporadically. A principal component analysis of the data matrix confirmed this order of relative influence of the test variables on the MIC. Since each fungal isolate responded differently to combinations of process variables in the test, we conclude that any unified method for antifungal susceptibility determination represents a compromise, rather than an idealized system.

Antifungal Agents↗
PubMed2000

Cardiolipin is a normal component of human plasma lipoproteins.

Anticardiolipin (anti-CL) antibodies, diagnostic for antiphospholipid antibody syndrome, are associated with increased risks of venous and arterial thrombosis. Because CL selectively enhances activated protein C/protein S-dependent anticoagulant activities in purified systems and because CL is not known to be a normal plasma component, we searched for CL in plasma. Plasma lipid extracts [chloroform/methanol (2:1, vol/vol)] were subjected to analyses by using TLC, analytical HPLC, and MS. A plasma lipid component was purified that was indistinguishable from reference CL (M:1448). When CL in 40 fasting plasma lipid extracts (20 males, 20 females) was quantitated by using HPLC, CL (mean +/- SD) was 14.9 +/- 3.7 microgram/ml (range 9.1 to 24.2) and CL was not correlated with phosphatidylserine (3.8 +/- 1.7 microgram/ml), phosphatidylethanolamine (64 +/- 20 microgram/ml), or choline-containing phospholipid (1,580 +/- 280 microgram/ml). Based on studies of fasting blood donors, CL (>/=94%) was recovered in very low density, low density, and high density lipoproteins (11 +/- 5.3%, 67 +/- 11.0%, and 17 +/- 10%, respectively), showing that the majority of plasma CL (67%) is in low density lipoprotein. Analysis of relative phospholipid contents of lipoproteins indicated that high density lipoprotein is selectively enriched in CL and phosphatidylethanolamine. These results shows that CL is a normal plasma component and suggest that the epitopes of antiphospholipid antibodies could include CL or oxidized CL in lipoproteins or in complexes with plasma proteins (e. g., beta(2)-glycoprotein I, prothrombin, protein C, or protein S) or with platelet or endothelial surface proteins.

Antibodies, Antiphospholipid↗
PubMed2000

Industrial emergency response training: an assessment of long-term impact of a union-based program.

BACKGROUND: The long-term impact of the United Automobile Workers' (UAW) Industrial Emergency Response Training on health and safety conditions and practices in plants is described. Two strategies are combined in this 24-hour training: the use of peer (worker) trainers and a participant-centered approach, the Small Group Activity Method (SGAM). METHODS: Impact was assessed through a telephone survey of 67 trainees conducted on average 16.8 months post-training. RESULTS: 74% of trainees reported using at least one of the provided written training materials after training. Of those reporting an accident in their facility after training, 88% said it was handled differently because of training. Trainees report that peer-trainers are more knowledgeable and communicate better than do other types of trainers. Trainees also reported that SGAM made training more applicable to their workplace. 98.5% of trainees like the methods utilized. CONCLUSIONS: Worker-trainers and SGAM are effective training methods enhancing the long-term impact of union-based workplace health and safety training programs.

Adult↗
PubMed2000

Multifactorial protective mechanisms to limit viral replication in the lung of mice during primary murine cytomegalovirus infection.

In this article, we investigated the protective host immune mechanisms against acute murine cytomegalovirus (MCMV) infection. For this purpose, we used various knockout mice lacking molecules, which include interferon-gamma (IFN-gamma), interferon-gamma receptor (IFN-gamma-R), interferon regulatory factor-1 (IRF-1), inducible nitric oxide synthase (iNOS), and perforin. We also used mutant mice lacking Fas molecule. When we infected these mice with MCMV and determined the viral titers in their lungs at different time points, we found that IFN-gamma, IFN-gamma-R, IRF-1, iNOS, and perforin-deficient mice developed significantly higher titers of infectious MCMV in the lung, compared to those observed in their respective wild-type controls. In the lungs of Fas-mutant mice, viral titers were similar to those obtained in wild-type mice.

Animals↗
PubMed2000

Prognostic value of protein C concentrations in neutropenic patients at high risk of severe septic complications.

OBJECTIVE: To assess the prognostic value of protein C, endogenous activated protein C, and D-dimer concentrations in patients at high risk of developing severe septic complications secondary to cytostatic chemotherapy. DESIGN: Prospective, comparative, single-center study. SETTING: Specialized ward for treating patients with acute leukemia and associated intensive care unit at a university hospital. SUBJECTS: Twenty-six consecutive patients who developed either severe sepsis (n = 13) or septic shock (n = 13) during chemotherapy-induced neutropenia (leukocytes <1,000/microL). INTERVENTION: None, other than standard care. MEASUREMENTS AND MAIN RESULTS: Baseline blood samples were obtained from 97 adult patients treated with intensive cytostatic chemotherapy. Serial blood sampling was performed in 62 of 97 patients who developed fever (>38.3 degrees C). Thirteen patients progressed to severe sepsis and 13 patients to septic shock. Protein C, endogenous activated protein C, and D-dimer were measured in these 26 patients. At fever onset, protein C concentrations decreased from normal baseline concentrations and were significantly lower in the group of patients who progressed to septic shock compared with those who developed severe sepsis (medians for protein C activity: 23.1% vs. 69.5%; p = .0003). The median elapsed time between detection of fever and the diagnosis of severe sepsis or septic shock was 16 hrs and 12 hrs, respectively. All septic shock patients died, whereas patients who progressed only to severe sepsis survived. CONCLUSIONS: Septic shock in neutropenic patients is associated with increased protein C consumption. The data demonstrate that the coagulation cascade is activated and produces a hypercoagulable state before the onset of clinical symptoms of severe sepsis and septic shock. Low protein C concentrations at the onset of fever and before the onset of clinical symptoms of severe sepsis or septic shock may have prognostic value in predicting an unfavorable outcome. Protein C measurements may help identify patients at risk in an early phase of neutropenic sepsis. It is also attractive to speculate that because low protein C concentrations were seen in these patients, protein C replacement may be beneficial in sepsis.

Antineoplastic Agents↗
PubMed2000

Causes of thrombophilia yet to be discovered: a personal view.

Prediction of genetic risk factors for venous thrombosis might best be left for only wise soothsayers. Nonetheless, based on the principle that hypercoagulability, either systemic or vascular-bed-specific, predisposes to thrombosis, we venture some speculations. Hyperactivity of platelets could be caused by elevated numbers of surface glycoproteins or defective signal transduction pathways. The reported efficacy of aspirin for prevention of venous thrombosis is consistent with the prediction that certain platelet defects may increase the risk of venous thrombosis. Hyperlipidaemia is associated with hypercoagulability, and lipoproteins exhibit procoagulant (e.g. triglyceride-rich particles and oxidized low-density lipoprotein) or anticoagulant (e.g. high-density-lipoprotein's cofactor activity for activated protein C/protein S) activities. This leads to the prediction that defects in lipids and/or lipoproteins may increase the risk for venous thrombosis. Interestingly, statins were recently reported to prevent the occurrence of venous thrombosis in the HERS (Heart and Estrogen/Progestin Replacement) trial. We also predict that new defects in the protein C pathway (e.g. defective endothelial protein C receptor or novel cofactors for activated protein C/protein S) will be discovered. Risk factors affecting the majority of patients will likely involve new single nucleotide polymorphisms (SNPs) like the factor V nt G1691A or prothrombin nt G20210A SNPs. The Human Genome Project will soon accelerate discovery of new SNPs that are risk factors for venous thrombosis.

Activated Protein C Resistance↗
PubMed1999

Extensive venous and arterial thrombosis associated with an inhibitor to activated protein C.

Activated protein C resistance (APCR) in the absence of alterations in the factor V gene has been observed during pregnancy, in patients on oral contraceptives, in the presence of antiphospholipid antibodies, and in patients with ischemic stroke. We report a 49-year-old woman with recurrent major venous and arterial thromboses who displayed pronounced APCR, yet no changes in the activated protein C (APC) cleavage sites of factor V. The APCR values determined by four different assays were similar to those obtained in plasma from a homozygote for factor V Q506. Addition of IgG isolated from the patient's serum to normal plasma lowered the APCR ratio from 2.4 to 1.6. Incubation of patient's IgG with normal APC resulted in a profound change in the mobility of APC in crossed immunoelectrophoresis. APC was also shown to bind to patient's IgG immobilized on a protein A agarose column. Factor Va inactivation by APC was inhibited by patient's IgG, but not by control IgG in the presence or absence of either phospholipids or protein S. These results provide evidence for the existence of an acquired antibody against APC in the patient's plasma, which gave rise to the APCR phenotype and was probably responsible for the major thrombotic events. We suggest that acquired APCR due to anti-APC antibodies be considered a potential cause for severe venous and arterial thromboses.

Autoantibodies↗
PubMed1999

Direct observation of the self-association of dilute proteins in the presence of inert macromolecules at high concentration via tracer sedimentation equilibrium: theory, experiment, and biological significance.

The technique of tracer sedimentation equilibrium [Rivas, G., et al. (1994) Biochemistry, 2341-2348 (1); Rivas, G., et al. (1996) J. Mol. Recognit. 9, 31-38 (2)] is utilized, together with an extension of the theory of sedimentation equilibrium of highly nonideal solutions [Chatelier and Minton, (1987) Biopolymers 26, 1097-1113 (3)], to characterize the thermodynamic activity and/or the state of association of a dilute, labeled macromolecular solute in the presence of an arbitary concentration of a second, unlabeled macromolecular solute. Experiments are performed on solutions of labeled fibrinogen (0.25-1 g/L) in bovine serum albumin (0-100 g/L) in the presence and absence of divalent cations (Ca(2+), Mg(2+)), and on solutions of labeled tubulin (0.2-0.6 g/L) in dextran (0-100 g/L). It is found that in the absence of the divalent cations, the large dependence of the thermodynamic activity of fibrinogen on BSA concentration is well accounted for by a simple model for steric repulsion. In the presence of the cations and sufficiently large concentrations of BSA (>30 g/L), fibrinogen appears to self-associate to a weight-average molar mass approximately twice that of monomeric fibrinogen. Tubulin appears to self-associate to an extent that increases monotonically with increasing dextran concentration, reaching a weight-average molar mass almost 3 times that of the alphabeta dimer in the presence of 100 g/L dextran. Possible biological ramifications are discussed.

Buffers↗
PubMed1999

Phenotypic and functional characterization of CD8(+) T cell clones specific for a mouse cytomegalovirus epitope.

A series of CD8(+) T cell clones, specific for the IE1 epitope YPHFMPTNL, of the immediate-early protein 1 of the murine cytomegalovirus (MCMV) were generated in order to determine their protective activity against this infection and correlate their phenotypic markers with antiviral activity. We found that the adoptive transfer of three of these anti-MCMV CD8(+) T cell clones into irradiated naive mice resulted in protection against challenge, while another CD8(+) T cell clone, of the same specificity, failed to confer protection. The clones that conferred protection against lethal challenge reduced greatly viral replication in the lung and other organs of the mice. Using one of the protective anti-MCMV CD8(+) T cell clones we found that in order to be fully protective the cells had to be transferred to recipient mice no later than 1 day after MCMV challenge. The adoptive transfer of these CD8(+) T cell clones also protected CD4(+) T-cell-depleted mice. Phenotypic characterization of the anti-MCMV clones revealed that the nonprotective clone expressed very low levels of CD8 molecules and produced only small amounts of TNF-alpha upon antigenic stimulation. Most importantly, our current study demonstrates that this MHC class I-restricted IE1 epitope of MCMV is efficiently presented to CD8(+) T cell clones in vivo and further strengthens the possibility of the potential use of CD8(+) T cell clones as immunotherapeutic tools against cytomegalovirus-induced disease.

Animals↗
PubMed1999

Phosphorylation- and activation-independent association of the tyrosine kinase Syk and the tyrosine kinase substrates Cbl and Vav with tubulin in B-cells.

Aggregation of the B-cell antigen receptor leads to the activation of the 72-kDa Syk protein-tyrosine kinase and the phosphorylation of tubulin on tyrosine. To explore the requirement of Syk catalytic activity for tubulin phosphorylation, tubulin was isolated from cytosolic fractions from anti-IgM-activated B-cells (DT40) that lacked endogenous Syk and immunoblotted with anti-phosphotyrosine antibodies. Tubulin was not tyrosine-phosphorylated in Syk- B-cells. Phosphorylation could be restored by the expression of wild-type, but not catalytically inactive, Syk. However, both catalytically inactive and wild-type Syk were capable of constitutive association with tubulin, indicating that tubulin phosphorylation is not required for this interaction. Anti-phosphotyrosine antibody immunoblotting of proteins adsorbed to colchicine-agarose revealed the presence of three major tubulin-associated phosphoproteins of 110, 90, and 74 kDa, the phosphorylation of which was dependent on Syk expression. The proteins of 110 and 90 kDa were identified as Cbl and Vav, two proto-oncogene products known to become prominently phosphorylated following receptor engagement. Both proteins were shown to be constitutively associated with tubulin.

Animals↗