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M Bastos

Publications and source records attributed to M Bastos.

At least 19 recordsLinked to original sources
PubMed2002

Studies on the properties of Celluclast/Eudragit L-100 conjugate.

A cellulase from Trichoderma reesei was immobilized on Eudragit L-100, a reversibly soluble polymer depending on the pH of the medium. The solubility of the modified cellulase was studied at different pH values. By changing the pH, the adsorption equilibrium of the derivatized proteins is switched towards the liquid phase, thus making recycling possible. This method allows for improved stability, without major loss of specific activity. The adsorption of cellulase on Eudragit lowers the enthalpy of denaturation, but affects only slightly the denaturation temperature. The use of carbodiimide was ineffective on linking the enzymes covalently to the polymer, since the immobilization process was found to be only mediated by non-covalent forces.

Adsorption↗
PubMed2002

[Childhood epilepsy syndromes and diseases].

The classification of seizures and epileptics syndromes made it possible on the prognostic level to differentiate the benign epilepsies and the serious epilepsies. In children, several concepts are to be considered for a better comprehension of the specificity of the epilepsies related to this age of the life: synaptogenesis, receptors ontogenesis and ionic channels, myelinogenesis, epigenesis. Epilepsy is often associated with cognitive disorders and behaviour disorders in children. The relations between cognitive functions and epilepsy are multifactorial with intrication of neurodevelopmental, environmental and psychogenic factors. Identification of epileptic syndromes genetically determined and genetic diseases becoming complicated by epilepsy, allow a complementary approach in the comprehension and diagnosis of childhood epilepsy.

Cerebral Cortex↗
PubMed2001

Thermodynamics of the helix-coil transition: Binding of S15 and a hybrid sequence, disulfide stabilized peptide to the S-protein.

Pancreatic ribonuclease A may be cleaved to produce two fragments: the S-peptide (residues 1-20) and the S-protein (residues 21-124). The S-peptide, or a truncated version designated as the S15 peptide (residues 1-15), combines with the S-protein to produce catalytically active complexes. The conformation of these peptides and many of their analogues is predominantly random coil at room temperature; however, they populate a significant fraction of helical form at low temperature under certain solution conditions. Moreover, they adopt a helical conformation when bound to the S-protein. A hybrid sequence, disulfide-stabilized peptide (ApaS-25), designed to stabilize the helical structure of the S-peptide in solution, also combines with the S-protein to yield a catalytically active complex. We have performed high-precision titration microcalorimetric measurements to determine the free energy, enthalpy, entropy, and heat capacity changes for the binding of ApaS-25 to S-protein within the temperature range 5-25 degrees C. The thermodynamic parameters for both the complex formation reactions and the helix-to-coil transition also were calculated, using a structure-based approach, by calculating changes in accessible surface area and using published empirical parameters. A simple thermodynamic model is presented in an attempt to account for the differences between the binding of ApaS-25 and the S-peptide. From this model, the thermodynamic parameters of the helix-to-coil transition of S15 can be calculated.

Amino Acids↗
PubMed1998

[Kidney transplantation in patients with type I and type II diabetes mellitus].

A total of 618 patients with end-stage renal disease received kidney transplants between 1980 and September 1996. Twenty eight of them were diabetics. Better results were achieved for type 1 diabetic patients than for type 2 (mortality: 5.9% vs 27.3%; functioning graft: 88.2% vs 72.7%). The morbility was also higher in those patients (infections: 81.8% vs 29.4%; vascular complications: 45.5% vs 17.6%). Actuarial patient and graft survival were lower for type 2 than for non diabetic patients. For type 1 diabetics the results are similar to those for non diabetics. Better results can probably be achieved by restricting the selection criteria. The decision to transplant or maintain on dialysis should be made on a case by case basis.

Adult↗
PubMed1997

[Craniocerebral imaging in children with short stature].

OBJECTIVE: To analyse the type and frequency of cranial CT and NMR imaging anomalies in children of short stature. PATIENTS AND INTERVENTIONS: We studied 57 children of short stature with a mean age (+/-SD) of 10.1 +/- 3.8 years, 34 boys and 23 girls, all of them with auxometric criteria of GH deficiency. After studying the pituitary function and determination of karyotype in the girls, the children were classified in to five groups:-Isolated GHD (IGHD) (n = 32), multiple pituitary hormone deficiency (MPHD) (n = 6), neurosecretory dysfunction (NSD) (n = 8), Turner syndrome (n = 7) and idiopathic short stature (ISS) (n = 4). The imaging methods used were cranial CT or NMR. RESULTS: Of the 57 children studied the CT/NMR was abnormal in 37(64.9%) children. We found anomalies in 65.6% of IGHD patients, 62.5% of NSD patients, 100% of MPHD patients and 57.1% and 25% in the Turner s. patients and ISS patients respectively. The most frequent anomaly was hypoplastic pituitary found in 50% of IGHD patients, 37.5% of NSD patients and 33.3% of MPHD patients. None of the cases of Turner s. or ISS had hypoplastic pituitary. An empty sella was the second most frequent anomaly found in 7 patients (IGHD-3, MPHD-3, DNS-1). Of the 25 children in which NMR was performed, 8 had hypoplastic pituitary and stalk and 2 had interruption of the pituitary stalk and ectopic neurohypophysis. CONCLUSION: These results strengthen the necessity for CT/NMR imaging in children of short stature which, besides allowing identification of tumors, also permits the diagnosis of idiopathic GHD because of its frequent association with cranial imaging anomalies, mainly hypoplastic pituitary.

Body Height↗
PubMed1995

Inhibitors of human heart chymase based on a peptide library.

We have synthesized two sets of noncleavable peptide-inhibitor libraries to map the S and S' subsites of human heart chymase. Human heart chymase is a chymotrypsin-like enzyme that converts angiotensin I to angiotensin II. The first library consists of peptides with 3-fluorobenzylpyruvamides in the P1 position. (Amino acid residues of substrates numbered P1, P2, etc., are toward the N-terminal direction, and P'1, P'2, etc., are toward the C-terminal direction from the scissile bond.) The P'1 and P'2 positions were varied to contain each one of the 20 naturally occurring amino acids and P'3 was kept constant as an arginine. The second library consists of peptides with phenylalanine keto-amides at P1, glycine in P'1, and benzyloxycarbonyl (Z)-isoleucine in P4. The P2 and P3 positions were varied to contain each of the naturally occurring amino acids, except for cysteine and methionine. The peptides of both libraries are attached to a solid support (pins). The peptides are evaluated by immersing the pins in a solution of the target enzyme and evaluating the amount of enzyme absorbed. The pins with the best inhibitors will absorb most enzyme. The libraries select the best and worst inhibitors within each group of peptides and provide an approximate ranking of the remaining peptides according to Ki. Through this library, we determined that Z-Ile-Glu-Pro-Phe-CO2Me and (F)-Phe-CO-Glu-Asp-ArgOMe should be the best inhibitors of chymase in this collection of peptide inhibitors. We synthesized the peptides and found Ki values were 1 nM and 1 microM, respectively. The corresponding Ki values for chymotrypsin were 10 nM and 100 microM. The use of libraries of inhibitors has advantages over the classical method of synthesis of potential inhibitors in solution: the libraries are reusable, the same libraries can be used with a variety of different serine proteases, and the method allows the screening of hundreds of compounds in short periods of time.

Amino Acid Sequence↗
PubMed1995

[Pancreatic transplantation].

The AA. report on their very limited experience with whole pancreatic transplantation. For the moment this is the only experience in Portugal, the detailed report of the 3 cases may be of some help to other Portuguese groups interested in starting this therapeutical approach for insulin-dependent end-stage renal failure patients. The main comments focus on the third simultaneous pancreas-kidney transplantation, which failed for technical reasons, mainly related to less than good selection of both donor and recipient. In all the three cases the technique preferred was the duodenocystostomy. The 2 first cases are doing very well, free of insulin and with normal glucose metabolism, at 15 and 7 months after grafting. The AA. also make some considerations on the indications, complications and follow-up of patients with pancreas-kidney transplantation.

Adult↗
PubMed1994

Heat conduction calorimeters: time constants, sensitivity and fast titration experiments.

Seven heat conduction calorimeters have been evaluated in terms of sensitivity and thermal response time. The use of a dynamic method to correct for the thermal inertia of the calorimeter is shown to reduce by one order of magnitude the time required to conduct a stepwise titration experiment involving a fast reaction. The ability to determine association constants of strong 1:1 complexes has been evaluated in terms of the precision of determining thermal energy.

Calorimetry↗
PubMed1994

[Pulse wave velocity as initial marker of atherosclerosis].

OBJECTIVE: To evaluate the influence of hypercholesterolaemia on arterial distensibility. MATERIAL AND METHODS: 43 male New Zealand White rabbits, with similar ages and weights, were included in the present study. The animals were divided in two groups: Group A (n = 15) was fed a normal diet; Group B (n = 28) was fed normal diet plus 0.1% cholesterol. at the beginning and after 6 and 9 months, blood samples were obtained for determination of serum cholesterol (total, esterified, LDL) and Triglyceride levels. Pulse wave velocity (PWV) was also evaluated, by mecanography, after 6 and 9 months of the beginning of the experiment. After 6 months (Group A = 4 and Group B = 7) and 9 months (Group A = 6 and Group B = 7) of the experiment, some animals were killed for anatomopathological studies. RESULTS: Major differences were obtained between the two groups, specially in what concerns to LDL and cholesterol levels (p < 0.001). There was also a remarkable difference in PWV between the two groups (6.078 +/- 0.162/9.002 +/- 0.196 m/s at 6 months and 7.639 +/- 0.590/9.557 +/- 0.543 m/s at 9 months) from the rabbits fed normal or cholesterol diet, respectively. The anatomical lesions were only significant after 9 months. However there was a decrease in aorta internal diameters at thoracic and renal levels at 6 months (34% and 53%) and at 9 months (29% and 33%), without significant changes in their thickness. In the heart, the left ventricle (LV) had a significant thickness increase after 6 months (about 43%). CONCLUSIONS: These data indicate that even before anatomical lesions had occurred, important functional changes are present, in the arterial wall. Then, the evaluation of the PWV could be a promising non-invasive diagnostic method of early atherosclerosis, with obvious implications concerning its prophylaxis and therapy.

Animals↗
PubMed1993

Effects of osmotic stress and growth stage on cellular pH and polyphosphate metabolism in Neurospora crassa as studied by 31P nuclear magnetic resonance spectroscopy.

High-resolution 31P-NMR was employed to investigate the effects of growth stage and environmental osmolarity on changes of polyphosphate metabolism and intracellular pH in intact Neurospora crassa cells. Our study showed that changes of these parameters were growth-dependent. The ratio of polyphosphate to orthophosphate in vacuoles increased from 2.4 to 13.5 in N. crassa as cells grew from early log phase to stationary phase. Cytoplasmic pH and vacuolar pH changed, respectively, from 6.91 and 6.49 in early log phase cells to 7.25 and 6.84 in stationary phase cells. Hypoosmotic shock of N. crassa produced growth-dependent changes including: (i) a rapid hydrolysis of polyphosphate with a concomitant increase in the concentration of the cytoplasmic phosphate, (ii) an increase in cytoplasmic pH, and (iii) an increase in vacuolar pH. Early log phase cells produced the most dramatic response whereas the stationary phase cells appeared to be recalcitrant to the osmotic stress. Thus, 95% and 60% of polyphosphate in the early log phase and mid-log phase cells, respectively, disappeared in response to hypoosmotic shock, but little or no hydrolysis of polyphosphate occurred in stationary cells. The cytoplasmic pH and the vacuolar pH increased in response to hypoosmotic shock by 0.4 and 0.53 unit, respectively, in early log phase cells; and by 0.22 and 0.27 unit, respectively, in the mid-log phase cells. In contrast, hypoosmotic shock of the stationary phase cells did not cause any change in intracellular pH. The osmotic stress-induced polyphosphate hydrolysis and pH changes in early log and mid-log phase cells were reversible, suggesting that these changes were related environment osmolarity. Addition of polyamines or basic amino acids which are known to be sequestered in vacuoles did not effect polyphosphate metabolism.

Amino Acids↗
PubMed1993

Unmodified pancreatic islet allograft rejection results in the preferential expression of certain T cell activation transcripts.

Polymerase chain reaction-assisted reverse transcription was used to study the temporal course of rejection in unmodified recipients of murine pancreatic islet cell allografts (DBA/2-->B6AF1) by using syngeneic tissues as controls. The histologic appearance of the grafts was analyzed in parallel. Preproinsulin and constant region of the TCR-beta chain transcripts were studied as markers of graft integrity and infiltrating T cell mass, respectively. The participation of certain cytokines and CTL were analyzed by the detection of IL-2, IFN-gamma, IL-4, and CTL-specific serine protease (granzyme B) transcripts. The time-related disappearance of intragraft preproinsulin transcripts correlated with graft destruction, whereas the intensity of intragraft TCR-beta chain transcript levels correlated with the magnitude of mononuclear leukocyte infiltration in allografts. In unmodified allografts, the magnitude of IL-2 and IFN-gamma intragraft mRNA levels correlated with the intense mononuclear leukocyte infiltrate found on histologic examination at day 8. Only after stable IL-2 gene transcription on day 8 does evidence of graft destruction become apparent, indicating that IL-2 gene activation is closely related to and probably required for expression of alloimmune cytopathic processes. In contrast, IL-4 transcripts were absent or detected in low copy number throughout this time course. Intragraft expression of granzyme B mRNA, a CTL-specific transcript, peaked from day 8 to day 12 in allografts compared with syngeneic grafts or normal tissue. In syngeneic grafts IL-2 and/or IL-4 mRNA was essentially not detected. Although IFN-gamma and granzyme B transcripts were detected in syngeneic grafts, after 4 days the levels of detected transcripts were far less than those noted in allografts. In vivo detection of intragraft IL-2 transcripts in the relative absence of detectable IL-4 transcripts strongly suggests IL-2-dependent immune effector mechanisms are associated with, and perhaps responsible, for allograft rejection. Apparently IL-4-dependent effector mechanisms are not necessary for allograft rejection.

Animals↗
PubMed1992

Enhanced tumor necrosis factor in anti-CD3 antibody stimulated diabetic NOD mice: modulation by PGE1 and dietary lipid.

Administration of OKT3 anti-CD3 monoclonal antibody (mAb) to patients for transplant rejection, is associated with a distinct and often severe clinical syndrome related to massive cytokine release. Previous reports have similarly demonstrated increased levels of serum tumor necrosis factor alpha (TNF alpha) in normal mice following administration of 1452-C11 anti-CD3 mAb. In this study, we compared serum TNF alpha levels at baseline and after anti-CD3 stimulation among three groups of mice: normal BALB/c controls, pre-diabetic non-obese diabetic (NOD) mice, and diabetic NOD mice. Baseline serum TNF alpha levels, as measured by L929 cell bioassay, were 2xhigher in diabetic NOD and 3xhigher in pre-diabetic NOD compared with BALB/c. Ninety minutes after anti-CD3 mAb stimulation, serum from BALB/c controls and pre-diabetic NOD contained 2- to 8-fold higher levels of TNF-alpha as compared to untreated control mice. In contrast, following anti-CD3 mAb, there was a dramatic 20-fold increase in serum TNF alpha in diabetic NOD mice (levels > 5000 pg/ml). Additionally, anti-CD3 mAb increased the steady-state TNF alpha mRNA transcripts. Spleens from diabetic mice given anti-CD3 mAb had higher steady-state TNF alpha mRNA than spleen from normal mice similarly treated. The enhanced release of circulating TNF alpha after anti-CD3 mAb in diabetic NOD mice was abrogated by pre-treatment of mice with prostaglandin E1 (PGE1) 30 min prior to anti-CD3 mAb stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Alprostadil↗
PubMed1992

[Pulse wave velocity as expression of arterial compliance and its importance in the evaluation of arterial hypertension].

STUDY OBJECTIVE: To determine the arterial compliance through the evaluation of pulse wave velocity. DESIGN: Open study with direct comparison of different groups within a 12 week evaluation period. PATIENTS: 69 patients, 49 with hypertension and 20 normals individuals. INTERVENTIONS: Different groups with the following treatments: Isradipine, Lisinopril, Dilevalol and no therapy. MEASUREMENTS AND MAIN RESULTS: There is marked differences in the pulse wave velocity when hypertensive patients are compared with normal individuals (p < 0.001). In a 12 week therapeutic evaluation there is an improvement in the pulse wave velocity particularly when the arterial pressure was lowered to normal values in the hypertensive patients: Lisinopril (p < 0.005), Isradipine (p < 0.005), Dilevalol (p < 0.025). CONCLUSIONS: It is very easy to evaluate the pulse wave velocity. Arterial compliance, which may be evaluated using the pulse wave velocity, is significantly reduced in hypertensive patients, compared with age-matched control subjects. The use of antihypertensive drugs is associated with changes in arterial compliance. There is a significant decrease in the pulse wave velocity after the administration of ACE inhibitors, calcium channel blockers and beta blockers for an equivalent fall in blood pressure. These observations emphasizes the potential importance of the changes in the large arteries, considered as either an associated factor or a consequence of elevated blood pressure, in the evaluation of the cardiovascular morbidity, and mortality of patients treated for hypertension.

Adult↗