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S Morais

Publications and source records attributed to S Morais.

At least 19 recordsLinked to original sources
PubMed2006

Dietary neutral lipid level and source in Senegalese sole (Solea senegalensis) larvae: effect on growth, lipid metabolism and digestive capacity.

Contrary to larval essential fatty acid (EFA) requirements, the effect of dietary neutral lipid supply has been little investigated in marine fish larvae. The present work investigates the effect of feeding Senegalese sole larvae on Artemia enriched with higher or lower doses of lipid emulsion. Two lipid sources - soybean oil and fish oil - were compared. From 16 days after hatching (DAH) onwards, larvae were fed one of four experimental treatments: Artemia enriched on a high or low dose of soybean oil emulsion (HS and LS) or Artemia enriched on a high or low dose of fish oil emulsion (HF and LF). In terms of growth, the dietary lipid level did not have a significant effect while the soybean oil treatments induced a lower growth than the fish oil-enriched Artemia. The fatty acid (FA) composition of the larvae closely reflected the dietary quantitative and qualitative FA profile. Only slight dietary effects were noted in the activity of trypsin, lipase and alkaline phosphatase. A higher amount of lipid droplets was noticeable in the posterior intestine epithelia and in the hepatocytes of larvae fed Artemia enriched with higher lipid doses, while LS-Artemia induced the lower lipid accumulation on the basal zone of the enterocytes, in accordance with the lowest total lipid level measured in this treatment. These results suggest an important effect of dietary total lipid level on lipid accumulation in the enterocytes and on FA absorption. At 33 DAH a tube feeding trial was conducted with 14C-labelled oleic acid (OA) or triolein (TRI), showing that the lower accumulation of lipid droplets in the larvae fed LS was associated with a significantly higher absorption and retention in the gut and body tissues of the TRI label. For OA no significant differences between treatments were found. TRI label was considerably more evacuated than OA, indicating that sole larvae may have a lower capacity to incorporate a triacylglycerol, which needs to be digested. Finally, OA appears to be preferentially utilized for energy production, accumulating more in larval tissues when absorbed in higher amounts.

Animals↗
PubMed2004

Dietary TAG source and level affect performance and lipase expression in larval sea bass (Dicentrarchus labrax).

The influence of dietary TAG source (fish oil, triolein, and coconut oil) and level (7.5 and 15% of the diet) on growth, lipase activity, and mRNA level was studied in sea bass larvae, from mouth opening until day 24 and from day 37 to 52. Fish oil and triolein induced better growth in both experiments, this being significant at a higher dietary level. Coconut oil significantly decreased growth at the higher level, possibly as the result of an excessive supply of medium-chain TAG. Growth was not related to lipase specific activity, suggesting a production in excess to dietary needs. Body lipid content was positively related to dietary lipid level and was affected by lipid quality. In addition, larval FA composition generally reflected that of the diet. The source of dietary lipid, but not the quantity, was shown to affect lipase activity significantly. Coconut oil diets induced the highest lipase activity, whereas the effect of fish oil was age dependent-it was similar to coconut oil at day 24 but induced the lowest lipase activity in 52-d-old larvae. The differential lipase response was probably caused by differences in the FA composition of the diet, related to the specificity of lipase toward FA differing in chain length and degree of saturation. No significant differences were found in lipase/glyceraldehyde-3-phosphate dehydrogenase mRNA, which suggests the existence of a posttranscriptional regulation mechanism.

Age Factors↗
PubMed2003

Metallothionein concentrations in a population of Patella aspera: variation with size.

The determination of metallothionein (MT) concentrations in species with different feeding habits is important from the ecotoxicological point of view because it provides a better understanding of the role of these proteins in metal uptake pathways. The main objective was study the variation of MT and metal (Cd, Cu and Zn) concentrations with size and weight in the limpets Patella aspera. In addition investigate the relationship between MT and metal concentrations in limpets from different metal load environments of the south coast of Portugal with the aim to use MT in P. aspera as a biomarker of metal exposure. MT concentrations in the whole soft tissues of P. aspera increased with size and weight while metals decrease with size and weight. MT concentrations showed no significant relationship with Cd or Cu concentrations in the limpets from the South Portuguese Coast. However, a negative exponential relationship detected between MT and Zn concentrations suggest that Zn bound to MT might be displaced by Cd or Cu ions. The ability of limpets to store both Cd and Cu bound to MT may be responsible for the tolerance of this species to contaminated environments. The relationship between MT concentrations and Zn indicates that MT seems to play a minor role in binding Zn in Patella species.

Adaptation, Physiological↗
PubMed2003

Female haemophiliac homozygous for the factor VIII intron 22 inversion mutation, with transcriptional inactivation of one of the factor VIII alleles.

Phenotypic expression of X-linked recessive disorders, including haemophilia A, is rare in females. This report describes a female with sporadic severe haemophilia A. The female patient and her family members were evaluated by coagulation assays. Visible detectable disturbance of X chromosome structure or number, as well as 2N von Willebrand disease, were excluded as possible explanations of the haemophilia A phenotype. Molecular studies, factor VIII (FVIII) intron 22 inversion mutation analysis showed that the severe haemophilia A phenotype is the result of a maternally inherited, distal, FVIII gene inversion and a paternally inherited de novo, also distal, FVIII gene inversion. Furthermore, comparative single-stranded conformation polymorphism analysis revealed the absence of detectable maternally inherited abnormal FVIII gene transcript in the patient's peripheral blood lymphocytes. X chromosome methylation analysis indicates that this could be explained by preferential inactivation of the maternally inherited X chromosome carrying the distal FVIII gene inversion.

Alleles↗
PubMed2001

Evaluation of enzyme-linked immunoassays for the determination of chloroacetanilides in water and soils.

Reliable and sensitive indirect ELISAs for the quantitative determination of metolachlor, alachlor, and acetochlor were developed. Each herbicide was conjugated to a carrier protein via thioether linkage, and the product was used either as an immunogen or to prepare coating conjugates. The suitability of using the same chemical strategy to raise polyclonal antibodies against chloroacetanilides structurally related compounds and their metabolites is discussed. Under best conditions, detection limits of 0.06, 0.3, and 0.4 microg/L for metolachlor, alachlor, and acetochlor were reached, respectively. The optimized ELISAs were also highly specific, showing little or no cross-reactivity to other similar compounds. Immunoassays were used as a toolto determine critical chloroacetanilide herbicides in water and soil samples without purification steps. The excellent recoveries obtained (mean value ranging between 90% and 98%) confirm the potential of this approach to control these herbicides in the environment being applied as a "screening" method either for field monitoring or laboratory.

Acetamides↗
PubMed2000

Application of stripping voltammetry and microelectrodes in vitro biocompatibility and in vivo toxicity tests of AISI 316L corrosion products.

Adsorptive stripping voltammetric procedures, using mercury film microelectrodes, were optimised and applied to quantify total iron, chromium and nickel in samples of osteoblast-like cells culture medium and mice organs (liver, kidney and spleen) obtained from, respectively, in vitro and in vivo 316L stainless steel corrosion products biocompatibility and toxicity studies. The methods were based on the pre-concentration of the iron-catechol complex by adsorption at the potential of -1.80 V (vs. Ag/AgCl), of the chromium-diethylenetriaminepentaacetic acid complex at -1.00 V or -1.15 V (vs. Ag/AgCl) and of the nickel-dimethylglyoxime complex at -0.70 V (vs. Ag/AgCl). The detection limits achieved for each metal ion (i) in the culture medium were 1.93x10(-8) mol/L Fe, 2.80x10(-10) mol/L Cr and 7.70x10(-9) mol/L Ni for a collection time of 30 s, 40 s and 10 s, respectively, and (ii) in the mice organ solutions were 1.37x10(-8) mol/L Fe, 1.54x10(-8) mol/L Cr and 1.58x10(-9) mol/L Ni for an adsorption time of 25 s, 25 s and 15 s, respectively. The accuracy of the proposed procedures was verified by comparison of the results obtained by adsorptive stripping voltammetry with those attained by atomic absorption spectrometry for the same set of samples and good agreement was found. The in vitro study showed that stainless steel corrosion products affect the expression of the osteogenic phenotype. The in vivo mice model, used to investigate the systemic effects provoked by the corrosion products per se, indicated that Fe, Cr and Ni are partially accumulated in the organs studied and that Ni induced the more significant morphological alterations.

Animals↗
PubMed1999

Immunofiltration: a methodology for preconcentration and determination of organic pollutants.

A new concentration procedure using an immunofiltration-based method is described. The approach enables quantitative determination of organic pollutants by filtering large volumes of sample through a poly(vinylidene difluoride) membrane where antibodies have been immobilized by passive adsorption. The analysis is based on a sequential competitive enzyme immunoassay. A wide range of sample volumes have been tested (0.2-5.0 mL) for each type of antibody. The improvement on the assay sensitivity and specificity achieved by means of this concentration procedure is discussed. Using this technique and the insecticide carbaryl as a model analyte, a concentration factor of at least 13 and a limit of detection of 4.75 ng/L are accomplished. The suitability of this methodology is demonstrated by the quantification of the insecticide in several types of water samples (bottled, estuarine, and physiological-saline solutions) with recoveries ranging between 102 and 111%. This method has proved to concentrate carbaryl directly, in an accurate way, for residue analysis without using organic solvents or any extraction process. Furthermore, this procedure offers the advantages of carrying out in the same system both preconcentration and quantitative determination of the analyte.

Antibodies↗
PubMed1999

Selection and characterisation of membranes by means of an immunofiltration assay. Application to the rapid and sensitive determination of the insecticide carbaryl.

The characterisation and selection of membranes by means of an immunofiltration assay is described. The chemical composition of the membranes was: nitro-cellulose, polyamide, polyvinylidene difluoride, polyethersulfone, cellulose acetate, regenerated cellulose, cellulose nitrate, and glass fibre. In order to characterise the membranes according to their binding capacity, immobilisation stability, sensitivity and hydrodynamic properties, two basic immunofiltration formats were performed. In both formats, enzyme label (horseradish peroxidase, HRP) and colorimetric detection were used. In the immobilised antibody format, three monoclonal antibodies (mAb) against the insecticide carbaryl were immobilised on the membranes by passive adsorption. In the immobilised hapten format, two haptens conjugated to bovine serum albumin (BSA) were immobilised. Immobilon-P was the best membrane with regard to the characterisation criteria and permitted the filtration of large volume (5.0 ml) through the membrane without release of the receptor. The immobilisation of the receptor (antibody or haptenic conjugate) was pH dependent. Good results with regard to mAb-antigen recognition, were obtained using 50 mM carbonate/bicarbonate buffer, pH 9.6. However, the most sensitive assays were achieved using, 10 mM phosphate buffer, 137 mM NaCl, 2.7 mM KCI (PBS), pH 7.4 as immobilisation buffer. Furthermore, all these results permit the choice of the best membrane for the rapid and sensitive determination of carbaryl. This study will assist the development of dipsticks, immunoelectrodes, membrane-based immunoreactors or immunoconcentration devices that are based on the use of membranes as immunosupports.

Animals↗
PubMed1999

In vitro osteoblastic differentiation of human bone marrow cells in the presence of metal ions.

For periods up to 21 days human bone marrow was cultured in control conditions that favor the proliferation and differentiation of osteoblastic cells. The effect of AISI 316L corrosion products and the corresponding major separate metal ions (Fe, Cr, and Ni) were studied in three different phases of the culture period in order to investigate the effects of metal ions in cell populations representative of osteoblastic cells in different stages of differentiation. Toxicity consequences of the presence of metal ions in bone marrow cultures were evaluated by biochemical parameters (enzymatic reduction of MTT, alkaline phosphatase activity, and total protein content), histochemical assays (identification of ALP-positive cells and Ca and phosphates deposits), and observation of the cultures by light and scanning electron microscopy. Culture media were analyzed for total and ionized Ca and P and also for metal ions (Fe, Cr, and Ni). The presence of AISI 316L corrosion products and Ni salt in bone marrow cultures during the first and second weeks of culture significantly disturbs the normal behavior of these cultures, interfering in the lag phase and exponential phase of cell growth and ALP expression. However, the presence of these species during the third week of culture, when expression of osteoblastic functions occurs (mineralization process), did not result in any detectable effect. Fe salt also disturbs the behavior of bone marrow cell cultures when present during the lag phase and proliferation phase, and a somewhat compromised response between the normal pattern (control cultures) and intense inhibition (AISI 316L corrosion products and Ni salt-added cultures) was observed. Fe did not affect the progression of the mineralization phase. Osteogenic cultures exposed to Cr salt (Cr3+) presented a pattern similar to the controls, indicating that this element does not interfere, in the concentration studied, in the osteoblastic differentiation of bone marrow cells. Quantification of metal ions in the culture media showed that Cr (originated from AISI 316L corrosion products but from not Cr3+ salt) and Ni (originated from AISI 316L corrosion products and Ni salt) appear to be retained by the bone marrow cultures.

Alkaline Phosphatase↗
PubMed1998

Decreased consumption of Ca and P during in vitro biomineralization and biologically induced deposition of Ni and Cr in presence of stainless steel corrosion products.

The purpose of this study was to investigate the effects of 316L stainless steel (SS) corrosion products on the in vitro biomineralization process, because tissue necrosis, bone loss, impaired bone mineralization, and loosening of orthopedic implants are associated with ions and debris resulting from biodegradation. Rat bone marrow cells were cultured in experimental conditions that favored the proliferation and differentiation of osteoblastic cells and were exposed to SS corrosion products obtained by electrochemical means for periods ranging from 1 to 21 days. Quantification of total and ionized Ca and P, as well as Fe, Cr, and Ni, ions in the culture media of control and metal added cultures during the incubation period was performed to study the influence of corrosion products on the Ca and P consumption that occurs during the mineralization process. Control cultures and metal effects on cultures were evaluated concerning DNA content, enzymatic reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and alkaline phosphatase (ALP) activity. Histochemical detection of ALP, Ca, and phosphate deposition, and examination of the cultures by scanning and transmission electron microscopy (SEM and TEM) were also performed. The presence of SS corrosion products resulted in impairment of the normal behavior of rat bone marrow cultures. Levels of Cr and Ni in the medium of cultures exposed to 316L SS corrosion products decreased throughout the incubation period, suggesting a regular deposition of these species; these results were supported by TEM observation of the cultures. Cultures exposed to the corrosion products presented lower DNA content, MTT reduction, and ALP activity and failed to form mineralized areas. These cultures showed negative staining on histochemical reactions for the identification of calcium and phosphate deposition and SEM and TEM examination did not show mineral globular structures or mineralization foci, respectively, which is characteristic of cultures grown in control conditions. These results suggest that metal ions associated with 316L SS are toxic to osteogenic cells, affecting their proliferation and differentiation.

Animals↗
PubMed1998

Effects of AISI 316L corrosion products in in vitro bone formation.

Rat bone marrow cells were cultured in experimental conditions that favour the proliferation and differentiation of osteoblastic cells (i.e., 2.52 x 10(-4) mol l(-1) ascorbic acid, 10(-2) mol l(-1) beta-glycerophosphate and 10(-8) mol l(-1) dexamethasone) in the absence and in the presence of stainless-steel corrosion products, for a period of 18 days. An AISI 316L stainless-steel slurry (SS) was obtained by electrochemical means and the concentrations of the major metal ions, determined by atomic absorption spectrometry, were 8.78 x 10(-3) mol l(-1) of Fe, 4.31 x 10(-3) mol l(-1) of Cr and 2.56 x 10(-3) mol l(-1) of Ni. Bone marrow cells were exposed to 0.01, 0.1 and 1% of the SS and at the end of the incubation period, control and treated cultures were evaluated by histochemical assays for the identification of the presence of alkaline phosphatase and also calcium and phosphate deposition. Cultures were further observed by scanning electron microscopy. Levels of total and ionised calcium and phosphorus in the culture media collected from control and metal exposed cell cultures were also quantified. Histochemical staining showed that control cultures presented a strong reaction for the presence of alkaline phosphatase and exhibited formation of calcium and phosphates deposits. The presence of 0.01% SS caused no detectable biological effects in these cultures, 0.1% SS impaired osteoblastic behaviour and, 1% SS resulted in cell death. In the absence of bone cells, levels of total and ionised calcium and phosphorus in the control and metal added culture medium were similar throughout the incubation period. A significant decrease in the levels of ionised calcium and phosphorus were observed in the culture medium of control cultures and also in cultures exposed to 0.01% SS after two weeks of incubation, an event related with the formation of mineral calcium phosphate deposits in these cultures. In cultures grown in the presence of 0.1 and 1% SS corrosion products, levels of calcium and phosphorus were similar to those observed in the absence of cells. Results showed that stainless-steel corrosion products above certain concentrations may disturb the normal behaviour of osteoblast-like rat bone marrow cell cultures.

Alkaline Phosphatase↗
PubMed1998

Chromium determination in osteoblast-like cell culture medium by catalytic cathodic stripping voltammetry with a mercury microelectrode.

A catalytic cathodic stripping voltammetric procedure for the determination of total chromium in osteoblast-like cell culture medium using a mercury film microelectrode (MFM) was optimised. The method is based on the pre-concentration of the Cr(III)-diethylenetriaminepentaacetic acid (DTPA) complex by adsorption at the potential of-1.00 V (vs. Ag/AgC1) in the presence of 10 x 10(-3) mol/L DTPA, 0.70 mol/L sodium nitrate, 0.04 mol/L sodium acetate and 1.0 x 10(-3) mol/L potassium permanganate at pH 5.9-6.0. The limit of detection obtained for a 40 s collection time was 2.80 x 10(-10) mol/L of chromium. The results achieved by stripping voltammetry using the MFM were compared to those obtained by atomic absorption spectrometry (AAS) to ensure the reliability of the electrochemical method. This procedure proved to be an alternative to AAS and valuable in biocompatibility studies performed in vitro using osteoblast-like cells.

Acetates↗
PubMed1997

A comparative study by the enzyme-linked immunofiltration assay of solid phases used in the development of flow immunosensors.

The application of an inert membrane-based, enzyme-linked immunofiltration assay (ELIFA) to the characterization of immunosorbents suitable for flow immunosensor development is described. For direct assays, eight monoclonal antibodies (MAb) raised against the insecticide carbaryl were immobilized on three sorbents, namely, controlled pore glass (CPG), hydrazide derivatized agarose beads and a hydrophilic polymer with immobilized Protein A/G. The interaction between immobilized antibodies and antigen was directly detected using a carbaryl hapten conjugated to horseradish peroxidase. Immunosorbent characterization was based on both sensitivity and re-usability. Optimal immunosorbent regeneration was achieved using 0.1 M glycine/HCl, pH 2.0 as the desorbent solution. The best covalent immunosorbent was obtained by immobilizing LIB-CNA36 MAb on hydrazide derivatized agarose beads. The best immunosorbent obtained by reversible immobilization was LIB-CNH45 MAb on Protein A/G. Using this support the eventual irreversible denaturation of covalently immobilized MAbs was overcome. For indirect assays, N-hydroxisuccinimide derivatized agarose beads and glutaraldehyde-activated CPG were used as sorbents for hapten immobilization via the amino groups of a carrier protein. In this format, antigen-MAb interactions were detected using a peroxidase-conjugated rabbit anti-mouse immunoglobulin. The highest sensitivity was achieved by LIB-CNH45 MAb in combination with derivatized agarose beads. All these results demonstrated the suitability of ELIFA as a fast, precise and easy-to-use technique for immunosorbent selection.

Biosensing Techniques↗
PubMed1995

Immunophenotypic characteristics of acute leukaemia after myelodysplastic syndromes.

PURPOSE: To analyse the immunophenotype of acute leukaemia (AL) after myelodysplastic syndromes (MDS) (MDS-AL) and to compare the immunophenotypic profile of acute myeloblastic leukaemia (AML) secondary to MDS (MDS-AML) with that of "de novo"-AML. PATIENTS AND METHODS: Twenty patients with MDS-AL and 29 patients with "de novo"-AML were studied. Morphocytochemical and flow cytometric studies were done in each case. RESULTS: All the MDS-AL studied displayed a myeloid phenotype (MDS-AML). The main difference between MDS-AML and "de novo"-AML was a significantly higher frequency of CD34 expression in the first group. Differences concerning the expression of other non-lineage related or myeloid-associated markers were not statistically significant, although the percentage of cases CD15(+) was lower in MDS-AML. The overall frequency of expression of lymphoid-associated markers was similar in both groups, T-cell markers being more frequently detected. CONCLUSIONS: Our findings support the usefulness of immunophenotyping studies to characterize MDS-AL and suggest some immunophenotyping differences between MDS-AML and "de novo"-AML which might have biological and prognostic significance.

Adult↗
PubMed1994

Analysis of the essential sequences of the factor VIII gene in twelve haemophilia A patients by single-stranded conformation polymorphism.

We report the analysis by single-stranded conformation polymorphism of the essential sequences of the factor VIII(FVIII) gene (total length about 14 kb) including the entire coding sequence, flanking intronic sequences and the putative regulatory sequences 5' to the gene, in twelve unselected haemophilia A patients of Portuguese origin. Direct sequencing of the fragments with an altered migration pattern led to the identification of the disease-producing mutations in five patients. Three of these mutations, namely a 1 bp insertion in a motif of eight consecutive A residues at codon 1439 (FVIIIPorto3); a C to T transition at codon 1966 (Arg-->Stop), found in an inhibitor-positive patient (FVIIIMontijo); and a G to A transition at codon 479 (Gly-->Arg; FVIIIPorto1), have been reported in other ethnic groups. The two novel mutations are the substitution of AG by GG at the 3' end of intron 4 (FVIIILisboa1) destroying the invariant splice acceptor sequence, and a G to A transition at codon 1948 resulting in an aspartic acid substitution for glycine (FVIIIPorto2).

Base Sequence↗
PubMed1992

The gel test: some problems and solutions.

The gel centrifugation test (GT) is a method of transfusion serology, based on the fact that, after centrifugation, unagglutinated red blood cells (RBC) pass easily through a gel, while agglutinated RBC do not. The introduction of the GT to our blood bank transfusion routine [strictly following the manufacturer's instructions (DiaMed ID Micro Typing System)] resulted in problems with the interpretation of the results. These were overcome after the introduction of modifications, which included: (1) the systematic use of 1% RBC suspensions; (2) the use of 50 microliters of 1% RBC suspensions and 25 microliters of serum in all tests; (3) the control of all negative indirect antiglobulin tests (IAT) and direct antiglobulin tests (DAT) by the addition of 50 microliters of a 1% IgG coated RBC suspension followed by centrifugation; and (4) the systematic use of saline-suspended RBC for ABO typing in patients with positive DAT.

Blood Group Antigens↗
PubMedDate not supplied

In vitro biomineralization by osteoblast-like cells. I. Retardation of tissue mineralization by metal salts.

The cytocompatibility of stainless steel 316L (SS 316L) corrosion products was investigated with particular focus on the dose- and time-effect of electrochemically dissolved SS and the corresponding separate metal ions on osteogenic bone marrow derived cells. Type AISI 316L stainless steel (Fe 63.9%, Cr 18.0%, Ni 12.5%, Mo 2.8%, Si 1.2%, Mn 1.6% and C 0.025%, weight for weight) was anodically dissolved in Hank's Balanced Salt Solution (HBSS) and diluted to the following concentrations: 500 microg ml(-1) of Fe, 122 microg ml(-1) of Cr and 101 microg ml(-1) of Ni, as estimated by atomic absorption spectrometry. Similarly, salt solutions containing 50 microg ml(-1) of Fe (FeCl3 x 6H2O), 122 microg ml(-1) of Cr (CrCl3 x 6H2O) or 101 microg ml(-1) of Ni (NiNO3) were prepared. All solutions were diluted 1:10(3), 1:10(4) and 1:10(5) and their effects on cell proliferation and function of rabbit bone marrow cells were studied up to 28 days of culture. Bone marrow cells (second subculture) were cultured in alpha-Minimal Essential Medium (alpha-MEM) supplemented with 10% fetal bovine serum 10(-8) mol l(-1) dexamethasone, 2.52 x 10(-4) mol l(-1) ascorbic acid and 10(-2) mol l(-1) beta-glycerophosphate. The osteoblast response to the presence of metal ions was evaluated by biochemical assays (enzymatic reduction of MTT for evaluation of cell viability/proliferation, and estimation of alkaline phosphatase (ALP) activity) and histochemical assays (identification of ALP positive cells and calcium and phosphates deposits). Results suggest a decrease in the expression of the osteoblast phenotype in the presence of ion and alloy solutions. Stainless steel corrosion products elicited slight effects but the corresponding metal ions produced pronounced effects on the osteoblast phenotype, namely an alteration in the levels and temporal expression of ALP and lower and retarded tissue mineralization ability.

Alkaline Phosphatase↗