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Biomedical subjects

L Oliveira

Publications and source records attributed to L Oliveira.

At least 37 records · Page 2Linked to original sources
PubMed2000

Effects of hepatocellular iron imbalance on nitric oxide and reactive oxygen intermediates production in a model of sepsis.

BACKGROUND/AIMS: In mammals iron homeostasis is most important, as imbalance of iron such as iron overload may lead to severe diseases. Recently, it has been shown that the iron regulatory protein-1 is partially controlled by nitric oxide and reactive oxygen intermediates, molecules frequently seen in inflammatory events. The aim of the present study was to investigate the effects of impaired iron homeostasis on the interaction of nitric oxide, and reactive oxygen intermediate formation in hepatocytes in a model of acute inflammation. METHODS: Hepatocytes isolated from Corynebacterium parvum (C parvum)-injected rats were used to examine the formation of nitrogen and oxygen intermediates by iron deprivation and iron overload in the presence of lipopolysaccharide. In addition, we investigated the RNA binding and aconitase activity of iron regulatory protein-1. RESULTS: In the present study we show that iron overload in lipopolysaccharide-treated C. parvum-primed hepatocytes downregulated the RNA binding of iron regulatory protein-1 and aconitase activity. Subsequently, we observed a reduced formation of nitrite/nitrate and S-nitrosothiols but an increased production of reactive oxygen species, and hepatocellular damage. Moreover, the addition of iron to cell cultures caused a further increase in cellular damage, a drop in the cellular glutathione pool, and an increase in peroxynitrite and hydroxyl-like radicals. In contrast, addition of deferoxamine (an iron chelator) to lipopolysaccharide-treated C. parvum-primed hepatocytes protected cells by stabilizing the GSH content, maintaining the nitric oxide formation, and by reducing Fenton oxidants. CONCLUSIONS: Our results show that the antioxidative effects of iron chelators prevent the formation of toxic Fenton oxidants in severe inflammatory events, which should be considered in the treatment of disorders characterized by an iron imbalance.

Animals↗
PubMed2000

Effects of some isoprostanes on the human umbilical artery in vitro.

1. Cumulative concentration-effect curves for the selective prostanoid TP receptor agonist U46619 and six isoprostanes were constructed in the human isolated umbilical artery. 2. All compounds except 8-iso-PGF3 alpha produced concentration-dependent contractions. The contractile response to the isoprostanes increased with each cumulative addition up to a point, after which subsequent addition reduced the contraction below the previous level. This 'downturn' in the concentration-effect curve did not occur with U46619. 3. The potencies of the compounds tested were as follows (pEC50 +/- s.e.mean): U46619, 6.7 +/- 0.2; 8-iso-PGE2, 6.5 +/- 0.1; 8-iso-PGF2 alpha, 5.8 +/- 0.2; 8-iso-PGE1, 5.4 +/- 0.1; 8-iso-PGF1 alpha, 5.0 +/- 0.1; 8-iso-PGF2 beta > 4.8; 8-iso-PGF3 alpha >> 4.8 (n = 4-17). Neither 8-iso-PGF2 beta nor 8-iso-PGF3 alpha at 44 microM had a significant effect on cumulative concentration-effect curves to U46619. 4. The selective TP receptor antagonist GR32191 (0.1 microM) caused rightward shifts in the concentration-effect curves to all the active compounds. pA2 values for GR32191 against U46619, 8-iso-PGE2, 8-iso-PGF2 alpha, 8-iso-PGE1 were 7.6 +/- 0.2, 9 +/- 1, 8.2 +/- 0.3 and 7.7 +/- 0.3, respectively (n = 4). 5. Neither N omega-nitro-L-arginine methyl ester (100 microM) nor the selective DP receptor antagonist BW A868C (50 nM) affected the complex concentration-effect curve to 8-iso-PGE2 (n = 3). 6. Stable contractions to U46619 (1-3 microM) were unaffected by anandamide at concentrations up to 60 microM.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed2000

Mutational analysis of the interaction of the N- and C-terminal ends of angiotensin II with the rat AT(1A) receptor.

1. The role of different residues of the rat AT(1A) receptor in the interaction with the N- and C-terminal ends of angiotensin II (AngII) was studied by determining ligand binding and production of inositol phosphates (IP) in COS-7 cells transiently expressing the following AT(1A) mutants: T88H, Y92H, G196I, G196W and D278E. 2. G196W and G196I retained significant binding and IP-production properties, indicating that bulky substituents in position 196 did not affect the interaction of AngII's C-terminal carboxyl with Lys(199) located three residues below. 3. Although the T88A mutation did not affect binding, the T88H mutant had greatly decreased affinity for AngII, suggesting that substitution of Thr(88) by His might hinder binding through an indirect effect. 4. The Y92H mutation caused loss of affinity for AngII that was much less pronounced than that reported for Y92A, indicating that His in that position can fulfil part of the requirements for binding. 5. Replacing Asp(278) by Glu caused a much smaller reduction in affinity than replacing it by Ala, indicating the importance of Asp's beta-carboxyl group for AngII binding. 6. Mutations in residues Thr(88), Tyr(92) and Asp(278) greatly reduced affinity for AngII but not for Sar(1) Leu(8)-AngII, suggesting unfavourable interactions between these residues and AngII's aspartic acid side-chain or N-terminal amino group, which might account for the proposed role of the N-terminal amino group of AngII in the agonist-induced desensitization (tachyphylaxis) of smooth muscles.

Amino Acid Sequence↗
PubMed1999

Thioredoxin activation of iron regulatory proteins. Redox regulation of RNA binding after exposure to nitric oxide.

Iron regulatory proteins (IRP1 and IRP2) are redox-sensitive RNA-binding proteins that modulate the expression of several genes encoding key proteins of iron metabolism. IRP1 can also exist as an aconitase containing a [4Fe-4S] cluster bound to three cysteines at the active site. We previously showed that biosynthesis of nitric oxide (NO) induces the transition of IRP1 from aconitase to apoprotein able to bind RNA. This switch is also observed when cytosolic extracts are exposed to NO donors. However, the activation of IRP1 under these conditions is far from maximal. In this study we examined the capacity of physiological reducing systems to cooperate with NO in the activation of IRP1. Cytosolic extracts from the macrophage cell line RAW 264.7 or purified IRP1 were incubated with NO donors and subsequently exposed to glutathione or to thioredoxin (Trx), a strong protein disulfide reductase. Trx was the most effective, inducing a 2-6-fold enhancement of the RNA binding activity of NO-treated IRP1. Furthermore, the effect of NO on IRP1 from cytosolic extracts was abolished in the presence of anti-Trx antibodies. We also studied the combined effect of NO and Trx on IRP2, which exhibits constitutive RNA binding activity. We observed an inhibition of IRP2 activity following exposure to NO donors which was restored by Trx. Collectively, these results point to a crucial role of Trx as a modulator of IRP activity in situations of NO production.

Animals↗
PubMed1999

Characterization of excitatory prostanoid receptors in the human umbilical artery in vitro.

1. 5-HT and the prostanoid TP receptor agonists, U46619 and I-BOP, constricted the human umbilical artery with pEC50 values of 7.3+/-0. 2, 6.7+/-0.1, and 7.3+/-0.2, respectively. The selective TP receptor antagonist, GR32191 (0.1 microM), shifted the concentration-effect curves to U46619 and I-BOP to the right, but had no effect on the response to 5-HT. 2. The natural prostaglandins, PGF2alpha and PGE2, caused concentration-dependent contraction with pEC50 values of 5.2+/-0.2 and 4.9+/-0.2, respectively. PGD2 was a partial agonist with a pEC50 of 5.24+/-0.03. GR32191 (0.1 microM) inhibited the responses to all of these compounds suggesting that they produce contraction by acting at TP receptors. 3. Sulprostone failed to elicit contraction in the human umbilical artery at concentrations up to 4.4 microM suggesting the absence of EP1 and EP3 receptors. Despite this, 17-phenyltrinor PGE2 and GR63799 both induced contraction at concentrations above 1 microM, but the effects were sensitive to GR32191 (0.1 microM). 4. Fluprostenol had no effect on the human umbilical artery at concentrations up to 17 microM suggesting the absence of FP receptors. Cloprostenol was ineffective in two tissues, but caused contraction in one tissue at the highest concentration tested (1.7 microM). However, this response was abolished in the presence of GR32191 (0.1 microM). 5. The effects of four TP receptor antagonists were assessed by global non-linear regression analysis. GR32191, SQ29548, SQ30741, and ICI192605 competitively inhibited responses to U46619 with pKb values of 8.0+/-0.1, 7.6+/-0.1, 7.0+/-0. 2 and 8.1+/-0.1, respectively. 6 These results suggest that the human umbilical artery functionally expresses TP receptors, but not EP1, EP2 or FP receptors.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1999

A low resolution model for the interaction of G proteins with G protein-coupled receptors.

A model is presented for the interaction between G proteins and G protein-coupled receptors. The model is based on the fact that this interaction shows little specificity and thus conserved parts of the G proteins have to interact with conserved parts of the receptors. These parts are a conserved negative residue in the G protein, a fully conserved arginine in the receptor and a series of residues that are not conserved but always hydrophobic like the hydrophobic side of the C-terminal helix of the G protein and the hydrophobic side of a helix in the C-terminal domain of the receptor. Other, mainly cytosolic, factors determine the specificity and regulation of this interaction. The relation between binding and activation will be shown. A large body of experimental evidence supports this model. Despite the fact that the model does not provide atomic resolution, it can be used to explain some experimental data that would otherwise seem inexplicable, and it suggests experiments for its falsification or verification.

Amino Acid Sequence↗
PubMed1999

Evaluation of electromyographic activity and heart rate responses to isometric exercise. The role played by muscular mass and type.

The purpose of the present study was to examine the relationship between the electromyographic (EMG) activity and heart rate (HR) responses induced by isometric exercise performed by knee extension (KE) and flexion (KF) in men. Fifteen healthy male subjects, 21 +/- 1.3 years (mean +/- SD), were submitted to KE and KF isometric exercise tests at 100% of maximal voluntary contraction (MVC). The exercises were performed with one leg (right or left) and with two legs simultaneously, for 10 s in the sitting position with the hip and knee flexed at 90 degrees. EMG activity (root mean square values) and HR (beats/min) were recorded simultaneously both at rest and throughout the sustained contraction. The HR responses to isometric exercise in KE and KF were similar when performed with one and two legs. However, the HR increase was always significantly higher in KE than KF (P < 0.05), whereas the EMG activity was higher in KE than in KF (P < 0.05), regardless of the muscle mass (one or two legs) involved in the effort. The correlation coefficients between HR response and the EMG activity during KE (r = 0.33, P > 0.05) and KF (r = 0.15, P > 0.05) contractions were not significant. These results suggest that the predominant mechanism responsible for the larger increase in HR response to KE as compared to KF in our study could be dependent on qualitative and quantitative differences in the fiber type composition found in each muscle group. This mechanism seems to demand a higher activation of motor units with a corresponding increase in central command to the cardiovascular centers that modulate HR control.

Adult↗
PubMed1999

Cellular immune responses to Mycobacterium tuberculosis in a patient with Takayasu's arteritis.

INTRODUCTION: Takayasu's arteritis (TA) is a disease of unknown aetiology, characterized histologically by an inflammatory cell infiltrate that affects all layers of the arterial wall. Its association with tuberculosis (TB) was described 50 years ago, based on the presence of Langhan's giant cells and granulomas similar to those found in tuberculous lesions. The presence of TB in patients with TA well as been reported in several studies as well as a positive tuberculous response, but these associations could be fortuitous in countries where TB is endemic. Recent studies have shown that patients with TA have a heightened humoral response to mycobacterial antigens including the 65 kDa fraction, a heat shock protein (HSP) that has also been found to be expressed in the arterial wall of patients with TA. The purpose of this study was to determine lymphoproliferative response and interferon-gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) stimulated by live Mycobacterium tuberculosis (Mtb) H37Rv and a panel of mycobacterial antigens, in the hope of contributing to a better understanding of the cellular immune responses to Tuberculosis in Takayasu's arteritis. MATERIAL AND METHODS: Standard lymphoproliferation tests and IFN-gamma determination (ELISA) were performed in a 47-year old black man who fulfilled criteria for TA and 10 healthy controls, BCG vaccinated, Mantoux positive. The following were used: Mtb H37Rv, Purified Protein Derivative (PPD), purified 30 kDa, recombinant M. bovis BCG 10 kDa, 38 kDa, 65 kDa, 70 kDa, Short Term-Culture Filtrate Proteins (ST-CFP), Mid Term-Culture Filtrate Proteins (MT-CFP) obtained from H37Rv and phytohemaglutinin (PHA) as mitogen for positive control. RESULTS: PBMC from the patient with TA when compared to the mean values of the 10 healthy donors showed decreased proliferation in response to all antigens, with the exception of 65 kDa. The TA patient showed a similar pattern of IFN-gamma production to that obtained with control donors, with the exception of higher IFN-gamma production in response to ST-CFP and MT-CFP. CONCLUSIONS: We have shown reactivity of peripheral lymphocytes to HSP 65 kDa and a trend towards higher production of IFN-gamma in response to ST-CFP and MT-CFP in a patient with TA. These facts, together with the already established heightened humoral response, strengthens the association between TB and TA. However, HSP 65 kDa is not specific to TB and we conclude that similar studies using lymphocytes obtained from the arterial wall of TA patients may help to clarify the role of mycobacterial infection in Takayasu's arteritis.

Antigens, Bacterial↗
PubMed1998

Converse modulation of IRP1 and IRP2 by immunological stimuli in murine RAW 264.7 macrophages.

Iron regulatory proteins (IRP1 and IRP2) are two cytoplasmic RNA-binding proteins that control iron metabolism in mammalian cells. Both IRPs bind to specific sequences called iron-responsive elements (IREs) located in the 3' or 5' untranslated regions of several mRNAs, in particular mRNA encoding ferritin and transferrin receptor. In this study, we followed in parallel the in vivo regulation of the two IRPs in physiologically stimulated macrophages. We show that stimulation of mouse RAW 264.7 macrophage-like cells increased IRP1 IRE binding activity 4-fold, whereas IRP2 activity decreased 2-fold 8 h after interferon-gamma/lipopolysaccharide treatment. Decrease in IRP2 was not due to nitric oxide (NO) production and did not require de novo protein synthesis. Our data therefore indicate that the two IRPs can be conversely regulated in response to the same stimulus. In addition, the effect of endogenously produced NO on IRP1 was further characterized in an activated macrophage/target cell system. We show that NO acts as an intercellular signal to increase IRP1 activity in adjacent cells. As the effect was detectable within 1 h and did not require de novo protein synthesis, this result supports a direct action of NO on IRP1.

Animals↗
PubMed1998

Pudendal neuropathy is predictive of failure following anterior overlapping sphincteroplasty.

PURPOSE: This study assessed the efficacy of anterior overlapping sphincteroplasty and parameters predictive of a successful outcome. METHODS: Clinical findings and physiologic investigations of female patients who underwent anterior overlapping sphincteroplasty for fecal incontinence between 1988 and 1996 were reviewed. The extent of sphincter damage was assessed at needle electromyography as the number of quadrants exhibiting decreased motor unit potentials. Prolonged pudendal nerve terminal motor latencies were those of greater than 2.2 ms. The size of the endoanal ultrasound defect was assessed as degrees circumference of the external sphincter in which viable muscle was absent. Patients were reviewed by telephone questionnaire and were asked to grade the outcome of their surgery as excellent or good (success) or fair or poor (failure). Incontinence was graded using a scoring system of 0 (perfect continence) to 20 (complete incontinence). RESULTS: There were 100 patients who had an overlapping sphincteroplasty; complete follow-tip information was obtained for 77 patients at a median of 24 (range, 2-96) months. The median age was 47 (range, 25-80) years and they had a median duration of incontinence of four (range, 0.1-39) years. Prior sphincteroplasty had been performed in 30 patients with a median of one (range, 1-7) operations. Investigations performed included electromyography (n = 49), pudendal nerve terminal motor latency (n = 71), endoanal ultrasound (n = 49), and manometry (n = 67). Sixty percent of patients had improved continence and 42 (55 percent) considered their surgery to have been successful as attested to by a significant decrease in their incontinence score (from 15.1 +/- 4.5 to 4.3 +/- 4.2; P < 0.0001). Neither patient age, parity, prior sphincteroplasty, cause or duration of incontinence, extent of electromyography damage, size of the endoanal ultrasound defect, nor any manometric parameter correlated with outcome. However, 62 percent of 59 patients with bilaterally normal pudendal nerve terminal motor latencies had a successful outcome compared with only 16.7 percent of 12 patients with unilateral or bilateral prolonged pudendal nerve terminal motor latencies (P < 0.01). CONCLUSION: Bilateral normal pudendal nerve terminal motor latencies are the only factors predictive of long-term success after overlapping sphincteroplasty.

Adult↗
PubMed1998

Residues Val254, His256, and Phe259 of the angiotensin II AT1 receptor are not involved in ligand binding but participate in signal transduction.

The role of the external third of helix VI of the angiotensin II (AII) AT1 receptor for the interaction with its ligand and for the subsequent signal transduction was investigated by individually replacing residues 252-256 by Ala, and residues 259 or 261 by Tyr, and permanently transfecting the resulting mutants to Chinese hamster ovary (CHO) cells. Binding experiments showed no great changes in affinity of any of the mutants for AII, [Sar1]-AII, or [Sar1, Leu8]-AII, but the affinity for the nonpeptide antagonist DuP753 was significantly decreased. The inositol phosphate response to AII was remarkably decreased in mutants V254A, H256A, and F259Y. These results indicate that AT1 residues Val254, His256, and Phe259 are not involved in ligand binding but participate in signal transduction. Based in these results and in others from the literature, it is suggested that, in addition to the His256 imidazole ring, the Phe259 aromatic ring interacts with the AII's Phe8, thus contributing to the signal-triggering mechanism.

Angiotensin II↗
PubMed1998

[Cellular immune response to mycobacterial antigens].

The authors studied the immunological response before and after beginning anti-tuberculous therapy, of a previously health, HIV negative, 22 year old black male, from the Republic of Cape Verde. The patient had multiple vertebral bony lesions associated to subcutaneous abscesses. As immunological markers of antigen recognition, we measured blastogenic and cytotoxic responses and gamma-IFN secretion towards 30 kD, 65 kD, filtrate proteins of M. tuberculosis, M. tuberculosis (H37Rv) and PPD cultures. To characterise the role of cytokines during infection, expression of mRNA for gamma-IFN, IL-4 and IL-10 was also analysed. A slight increase of lymphocyte proliferation and gamma-IFN production was seen in response to purified protein derivative (PPD) and short term culture filtrate proteins (ST-CFP), after one month of therapy. More significant, was the increase in M. tuberculosis and PPD-specific cytolytic T lymphocyte response after one one month of treatment. After 6 months of treatment, blastogenic and cytotoxic responses and gamma-IFN production were considerably higher toward the antigen panel. The CD4/CD8 ratio increased from 0.7 to 1.4 after treatment. We observed that ST-CFP and MT-CFP induced increasingly higher lymphoproliferation and gamma-IFN production, confirming their role in the protective immune responses to M. tuberculosis. The reduced immune responses in the peripheral blood of this patient probably reflect a high activity in the local sites of infection. This case of disseminated tuberculosis infection maybe related to nutritional or social factors or may represent an example of reduced in ate resistance against tuberculosis infection.

Adult↗
PubMed1998

[Pulmonary embolism: a clinical case documented, in acute setting, using pulmonary scintigraphic perfusion].

The acute clinical evolution is presented of a patient with previous myocardial infarction who was admitted for dyspnea and chest pain. Acknowledging the possibility of a new ischemic event, confronted with aortic dissection or pulmonary embolism, the clinical investigation is described up to the scintigraphic diagnosis of pulmonary embolism. The clinical presentation, diagnostic investigation and therapeutic decisions are discussed, with emphasis on the usefulness of pulmonary scintigraphy.

Adult↗
PubMed1998

The interaction of class B G protein-coupled receptors with their hormones.

In common with many G protein-coupled receptors, dysfunction in members of the Class B or glucagon-like receptors can elicit a wide spectrum of disease related activities. Consequently, they are potential targets in many different areas of pharmacological research. Unlike the class A or rhodopsin-like receptors, for which at least some structural similarity to bacteriorhodopsin has been detected, absolutely no structural information is available for the Class B G protein-coupled receptors. We present a computational study that exploits the experimental work performed by evolution to indicate residues that are potentially involved in ligand binding in the Class B G protein-coupled receptors. We perform an analysis of mutations that occurred in a correlated fashion between the receptors and their peptidic ligands. The inference that the residues detected in this manner are involved in a direct interaction between the receptor and the ligand is in good agreement with the mutation studies that have already been published.

Amino Acid Sequence↗
PubMed1997

Mechanical bowel preparation for elective colorectal surgery. A prospective, randomized, surgeon-blinded trial comparing sodium phosphate and polyethylene glycol-based oral lavage solutions.

AIM: The aim of this study was to compare the cleansing ability, patient compliance, and safety of two oral solutions for elective colorectal surgery. METHODS: All eligible patients were prospectively randomized to receive either 4 l of standard polyethylene glycol (PEG) solution or 90 ml of sodium phosphate (NaP) as mechanical bowel preparation for colorectal surgery. A detailed questionnaire was used to assess patient compliance. In addition, the surgeons, blinded to the preparation, intraoperatively evaluated its quality. Postoperative septic complications were also assessed. The calcium serum level was monitored before and after bowel preparation. Statistical analysis was performed using the Wilcoxon's rank-sum test and Fisher's exact test. RESULTS: Two hundred patients, well matched for age, gender, and diagnosis, were prospectively randomized to receive either PEG or NaP solutions for elective colorectal surgery. All patients completed all phases of the trial. There was a significant decrease in serum calcium levels after administration of both NaP (mean, 9.3-8.8 mg/dl) and PEG (9.2-8.9 mg/dl), respectively (P < 0.0001), with no clinical sequelae. However, patient tolerance to NaP was superior to PEG: less trouble drinking the preparation (17 vs. 32 percent; P < 0.0002), less abdominal pain (12 vs. 22 percent; P = 0.004), less bloating (7 vs. 28 percent), and less fatigue (8 vs. 17 percent), respectively. Additionally, 65 percent of patients who received the NaP preparation stated they would repeat this preparation again compared with only 25 percent for the PEG group (P < 0.0001). Ninety-five percent of patients who received the NaP solution tolerated 100 percent of the solution compared with only 37 percent of the PEG group (P < 0.0001). For quality of cleansing, surgeons scored NaP as "excellent" or "good" in 87 compared with 76 percent after PEG (P = not significant). Rates of septic and anastomotic complications were 1 percent and 1 percent for NaP and 4 percent and 1 percent for PEG, respectively (P = not significant). CONCLUSION: Both oral solutions proved to be equally effective and safe. However, patient tolerance of the small volume of NaP demonstrated a clear advantage over the traditional PEG solution.

Colonic Diseases↗