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Biomedical subjects

L Oliveira

Publications and source records attributed to L Oliveira.

At least 19 recordsLinked to original sources

Comparative study of the whisky aroma profile based on headspace solid phase microextraction using different fibre coatings.

A dynamic headspace solid-phase microextraction (HS-SPME) and gas chromatography coupled to ion trap mass spectrometry (GC-(IT)MS) method was developed and applied for the qualitative determination of the volatile compounds present in commercial whisky samples which alcoholic content was previously adjusted to 13% (v/v). Headspace SPME experimental conditions, such as fibre coating, extraction temperature and extraction time, were optimized in order to improve the extraction process. Five different SPME fibres were used in this study, namely, poly(dimethylsiloxane) (PDMS), poly(acrylate) (PA), Carboxen-poly(dimethylsiloxane) (CAR/PDMS), Carbowax-divinylbenzene (CW/DVB) and Carboxen-poly(dimethylsiloxane)-divinylbenzene (CAR/PDMS/DVB). The best results were obtained using a 75 microm CAR/PDMS fibre during headspace extraction at 40 degrees C with stirring at 750 rpm for 60 min, after saturating the samples with salt. The optimised methodology was then applied to investigate the volatile composition profile of three Scotch whisky samples--Black Label, Ballantines and Highland Clan. Approximately seventy volatile compounds were identified in the these samples, pertaining at several chemical groups, mainly fatty acids ethyl esters, higher alcohols, fatty acids, carbonyl compounds, monoterpenols, C13 norisoprenoids and some volatile phenols. The ethyl esters form an essential group of aroma components in whisky, to which they confer a pleasant aroma, with "fruity" odours. Qualitatively, the isoamyl acetate, with "banana" aroma, was the most interesting. Quantitatively, significant components are ethyl esters of caprilic, capric and lauric acids. The highest concentration of fatty acids, were observed for caprilic and capric acids. From the higher alcohols the fusel oils (3-methylbutan-1-ol and 2.phenyletanol) are the most important ones.

Alcoholic Beverages↗

Effects of age and physical activity on the autonomic control of heart rate in healthy men.

The effects of the aging process and an active life-style on the autonomic control of heart rate (HR) were investigated in nine young sedentary (YS, 23 +/- 2.4 years), 16 young active (YA, 22 +/- 2.1 years), 8 older sedentary (OS, 63 +/- 2.4 years) and 8 older active (OA, 61 +/- 1.1 years) healthy men. Electrocardiogram was continuously recorded for 15 min at rest and for 4 min in the deep breathing test, with a breath rate of 5 to 6 cycles/min in the supine position. Resting HR and RR intervals were analyzed by time (RMSSD index) and frequency domain methods. The power spectral components are reported in normalized units (nu) at low (LF) and high (HF) frequency, and as the LF/HF ratio. The deep breathing test was analyzed by the respiratory sinus arrhythmia indices: expiration/inspiration ratio (E/I) and inspiration-expiration difference (deltaIE). The active groups had lower HR and higher RMSSD index than the sedentary groups (life-style condition: sedentary vs active, P < 0.05). The older groups showed lower HFnu, higher LFnu and higher LF/HF ratio than the young groups (aging effect: young vs older, P < 0.05). The OS group had a lower E/I ratio (1.16) and deltaIE (9.7 bpm) than the other groups studied (YS: 1.38, 22.4 bpm; YA: 1.40, 21.3 bpm; OA: 1.38, 18.5 bpm). The interaction between aging and life-style effects had a P < 0.05. These results suggest that aging reduces HR variability. However, regular physical activity positively affects vagal activity on the heart and consequently attenuates the effects of aging in the autonomic control of HR.

Adult↗

Identification of anaerobic threshold using heart rate response during dynamic exercise.

The objective of the present study was to characterize the heart rate (HR) patterns of healthy males using the autoregressive integrated moving average (ARIMA) model over a power range assumed to correspond to the anaerobic threshold (AT) during discontinuous dynamic exercise tests (DDET). Nine young (22.3 +/- 1.57 years) and 9 middle-aged (MA) volunteers (43.2 +/- 3.53 years) performed three DDET on a cycle ergometer. Protocol I: DDET in steps with progressive power increases of 10 W; protocol II: DDET using the same power values as protocol 1, but applied randomly; protocol III: continuous dynamic exercise protocol with ventilatory and metabolic measurements (10 W/min ramp power), for the measurement of ventilatory AT. HR was recorded and stored beat-to-beat during DDET, and analyzed using the ARIMA (protocols I and II). The DDET experiments showed that the median physical exercise workloads at which AT occurred were similar for protocols I and II, i.e., AT occurred between 75 W (116 bpm) and 85 W (116 bpm) for the young group and between 60 W (96 bpm) and 75 W (107 bpm) for group MA in protocols I and II, respectively; in two MA volunteers the ventilatory AT occurred at 90 W (108 bpm) and 95 W (111 bpm). This corresponded to the same power values of the positive trend in HR responses. The change in HR response using ARIMA models at submaximal dynamic exercise powers proved to be a promising approach for detecting AT in normal volunteers.

Adult↗

Effects of chronic heart disease on skeletal muscle fiber size.

Size changes in muscle fibers of subjects with chronic heart disease (CHD) have been reported, although a consensus has not been achieved. The aims of the present study were to investigate a possible association between CHD and fiber size changes in the brachial biceps compared to subjects without heart disease. Forty-six muscle samples were obtained in autopsies of individuals (13 to 84 years) without neuromuscular disorders, 19 (10 males and 9 females) with, and 27 (14 males and 13 females) without CHD. In all cases muscle sections were stained with hematoxylin and eosin and processed for the visualization of myofibrillar ATPase activity. The lesser diameter of type 1 and type 2 fibers was obtained tracing their outlines (at least 150 fibers of each type per sample) onto an image analyzer connected to a computer. The results were analyzed statistically comparing males and females with and without CHD. Type 1 fiber mean lesser diameters were 51.51 and 54.52 microm in males (normal range 34-71 microm) and 45.65 and 55.42 microm in females (normal range 34-65 microm) without and with CHD, respectively; type 2 fibers measured 54.31, 58.23, 41.15, and 49.57 microm, respectively (normal range 36-79 microm for males and 32-59 microm for females). No significant difference in fiber size was detected in 24 males with and without CHD, while in 22 females there was a significant increase in size in those with cardiomyopathy. We concluded that CHD does not determine significant changes in fiber size. However, in females, there is some hypertrophy which, despite within normal range, may reflect morphologic heterogeneity of the sample, or the daily life activities in the upper limbs as a compensatory mechanism to fatigability that affect predominantly the lower limbs in subjects with CHD.

Adolescent↗

Occurrence and characterization of a nucleopolyhedrovirus from Spodoptera littoralis (Lepidoptera: Noctuidae) isolated in the azores.

A nucleopolyhedrovirus (SpliMNPV-Az) was isolated from diseased larvae of Spodoptera littoralis, collected at the Island of S. Miguel in Azores. The virulence of this isolate was tested against S. littoralis larvae in laboratory. LD50 against 2nd and 3rd instars were not significantly different, 1.44x10(4), 3.89x10(4) OBs per larvae, respectively, but both were significantly different from that against 4th instar, which was 61.3x10(4) OBs per larvae. The complete codons sequence of SpliMNPV-Az Polh gene obtained was 750 bp (NCBI GenBank Accession No. AY600451). This sequence was compared to other 38 polyhedrin genes from NPVs and to 6 granulin genes from GVs and resulted to be identical to the sequence of a SpliMNPV previously published, thus indicating that the natural host of SpliMNPV-Az must be S. littoralis. Genetic distances estimated from restriction enzymes profiles showed SpliMNPV-Az is close to the Egyptian SpliMNPV type B, despite some degree of genetic divergence suggested by slight differences observed on PstI profile.

Amino Acid Sequence↗

Heavier-than-air flying machines are impossible.

Many G protein-coupled receptor (GPCR) models have been built over the years. The release of the structure of bovine rhodopsin in August 2000 enabled us to analyze models built before that period to learn more about the models we build today. We conclude that the GPCR modelling field is riddled with 'common knowledge' similar to Lord Kelvin's remark in 1895 that "heavier-than-air flying machines are impossible", and we summarize what we think are the (im)possibilities of modelling GPCRs using the coordinates of bovine rhodopsin as a template. Associated WWW pages: www.gpcr.org/articles/2003_mod

Amino Acid Sequence↗

Behavioral modulation by mutilation pictures in women.

Previous studies have shown that women are more emotionally expressive than men. It is unclear, however, if women are also more susceptible to the emotional modulation of behavior imposed by an affective stimulus. To investigate this issue, we devised a task in which female subjects performed six sequential trials of visual target detection following the presentation of emotional (mutilation and erotic) or neutral pictures (domestic utensils and objects) and compared the data obtained in the present study with those described in a previous study with male subjects. The experiment consisted of three blocks of 24 pictures and each block had an approximate duration of 4 min. Our sample consisted of 36 subjects (age range: 18 to 26 years) and each subject performed all blocks. Trials following the presentation of mutilation pictures (283 ms) had significantly slower reaction times than those following neutral (270 ms) pictures. None of the trials in the "pleasant block" (271 ms) was significantly different from those in the "neutral block". The increase in reaction time observed in the unpleasant block may be related in part to the activation of motivational systems leading to an avoidance behavior. The interference effect observed in this study was similar to the pattern previously described for men. Thus, although women may be more emotionally expressive, they were not more reactive to aversive stimuli than men, as measured by emotional interference in a simple reaction time task.

Adolescent↗

Contributions of stimulus valence and arousal to visual activation during emotional perception.

Neuroimaging experiments have revealed that the visual cortex is involved in the processing of affective stimuli: seeing emotional pictures leads to greater activation than seeing neutral ones. It is unclear, however, whether such differential activation is due to stimulus valence or whether the results are confounded by arousal level. In order to investigate the contributions of valence and arousal to visual activation, we created a new category of "interesting" stimuli designed to have high arousal, but neutral valence, and employed standard neutral, unpleasant, and pleasant picture categories. Arousal ratings for pleasant and neutral pictures were equivalent, as were valence ratings for interesting and neutral pictures. Differential activation for conditions matched for arousal (pleasant vs neutral) as well as matched for valence (interesting vs neutral) indicated that both stimulus valence and arousal contributed to visual activation.

Adult↗

Correlated mutation analyses on very large sequence families.

The 'omics era' (the era of genomics, proteomics, and so forth) is marked by a flood of data that need to be interpreted to become useful information. Thanks to genome sequencing projects, large numbers of sequence families with more than a thousand members each are now available. Novel analytical techniques are needed to deal with this avalanche of sequence data. Sequence entropy is a measure of the information present in an alignment, whereas sequence variability represents the mutational flexibility at a particular position. Entropy versus variability plots can reveal the roles of groups of residues in the overall function of a protein. Such roles can be as part of the main active site, part of a modulator binding site, or transduction of a signal between those sites. Residues that are involved in a common function tend to stay conserved as a group, but when they mutate, they tend to mutate together. Correlated mutation analysis can detect groups of residue positions that show this behaviour. The combination of entropy, variability and correlation is a powerful tool to convert sequence data into useful information. This analysis can, for example, detect the key residues involved in cooperativity in globins, the switch regions in ras-like proteins and the calcium binding and signalling residues in serine proteases. We have extrapolated from these three classes of structurally and functionally well-described proteins to G-protein-coupled receptors (GPCRs). We can detect the residues in the main functional site in GPCRs that are responsible for G-protein coupling, the residues in the endogenous agonist binding site, and the residues in between that transduce the signal to and fro between these sites. The results are discussed in the light of a simple two-step evolutionary model for the development of functional proteins.

Amino Acid Sequence↗

Aluminum toxicity studies in Vaucheria longicaulis var. macounii (Xanthophyta, Tribophyceae). I. Effects on cytoplasmic organization.

Using differential interference contrast (DIC) and epifluorescence microscopy, we tested the hypothesis that exposure to environmentally significant levels of aluminum (Al) would cause rapid changes in cytoplasmic organization in vegetative filaments of the coenocytic alga, Vaucheria longicaulis Hoppaugh var. macounii Blum resulting in the loss of cytoplasmic streaming. In untreated cells, DIC microscopy revealed the presence of cortical cytoplasmic strands that were oriented longitudinally to the cell axis as well as sub-cortical cytoplasmic strands that exhibited a reticulate morphology. Organelles such as chloroplasts and mitochondria translocated throughout the cell in close association with the cortical longitudinal cytoplasmic strands. Staining with the lipophilic dye, 3,3-dihexyloloxacarbocyanine, revealed structures that appeared to be endoplasmic reticulum (ER). This organelle closely resembled, in location and appearance, the cytoplasmic strands visualized using DIC microscopy. The addition of Al (80 µM) resulted in the inhibition of cytoplasmic streaming as well as the dissipation of the putative cortical longitudinal ER within one minute. Subsequently, the DIC-visible cortical cytoplasmic strands exhibited progressive degrees of disorganization. Throughout these changes, chloroplasts and mitochondria remained visibly associated with the cortical cytoplasmic strands.

Journal Article↗

Aluminum toxicity studies in Vaucheria longicaulis var. macounii (Xanthophyta, Tribophyceae). II. Effects on the F-actin array.

In this study, we test the hypothesis that exposure to environmentally significant concentrations of aluminum (Al, 80 µM) causes the microfilament array of Vaucheria longicaulis var. macounii vegetative filaments to become fragmented and disorganized. Changes in F-actin organization following treatment of vegetative filaments by Al are examined using vital staining with fluorescein phalloidin. In the cortical cytoplasm of the apical zone of pH 7.5 and pH 4.5 control cells, axially aligned bundles of F-actin lead to a region of diffuse, brightly stained material. Dimly stained focal masses are noted deeper in the cytoplasm of the apical zone whereas they are absent from the zone of vacuolation. The F-actin array is visualized in the cortical cytoplasm of the region of the cell, distal to the apical tip, which exhibits vigorous cytoplasmic streaming (zone of vacuolation) as long, axially aligned bundles with which chloroplasts and mitochondria associate. Thirty minutes following treatment with aluminum, and for the next 8-16 h, the F-actin array is progressively disorganized. The longitudinally aligned F-actin array becomes fragmented. Aggregates of F-actin, such as short rods, amorphous and stellate F-actin focal masses, curved F-actin bundles and F-actin rings replace the control array. Each of these structures may occur in association with chloroplasts or independently with no apparent association with organelles. Images are recorded which indicate that F-actin rings not associated with organelles may self-assemble by successive bundling of F-actin fragments. The fragmentation and bundling of F-actin in cells of V. longicaulis upon treatment with aluminum resembles those reported after diverse forms of cell disturbance and supports the hypothesis that aluminum-induced changes in the F-actin array may be a calcium-mediated response to stress.

Journal Article↗

Heart rate variability under resting conditions in postmenopausal and young women.

The aim of the present study was to compare the modulation of heart rate in a group of postmenopausal women to that of a group of young women under resting conditions on the basis of R-R interval variability. Ten healthy postmenopausal women (mean +/- SD, 58.3 +/- 6.8 years) and 10 healthy young women (mean +/- SD, 21.6 +/- 0.82 years) were submitted to a control resting electrocardiogram (ECG) in the supine and sitting positions over a period of 6 min. The ECG was obtained from a one-channel heart monitor at the CM5 lead and processed and stored using an analog to digital converter connected to a microcomputer. R-R intervals were calculated on a beat-to-beat basis from the ECG recording in real time using a signal-processing software. Heart rate variability (HRV) was expressed as standard deviation (RMSM) and mean square root (RMSSD). In the supine position, the postmenopausal group showed significantly lower (P<0.05) median values of RMSM (34.9) and RMSSD (22.32) than the young group (RMSM: 62.11 and RMSSD: 49.1). The same occurred in the sitting position (RMSM: 33.0 and RMSSD: 18.9 compared to RMSM: 57.6 and RMSSD: 42.8 for the young group). These results indicate a decrease in parasympathetic modulation in postmenopausal women compared to young women which was possibly due both to the influence of age and hormonal factors. Thus, time domain HRV proved to be a noninvasive and sensitive method for the identification of changes in autonomic modulation of the sinus node in postmenopausal women.

Adult↗

[Mitral flow propagation velocity assessed with M-mode color Doppler in patients with dilated cardiomyopathy].

BACKGROUND: The role of diastolic disfunction in the setting of severe left ventricular (LV) disfunction is usually forgotten due to difficulties in quantification and interpretation. The propagation velocity (PV) of mitral inflow into the left ventricle has been proposed as a load independent indicator of LV diastolic function. Our aim was to correlate PV with age, body surface, chambers dimensions, wall thickness and with classic parameters of systolic and diastolic function. We had in mind the validation of this index in the quantification of LV filling and in risk stratification of patients with dilated cardiomyopathy (DC). MATERIAL AND METHODS: We prospectively studied 32 consecutive patients, mean age 56.1 +/- 15.7 years, 22 (68.75%) male, mean ejection fraction 28.1 +/- 7.5%. All of them were in NYHA class < or = II and on ACE inhibitors. Eight (25%) were on beta-blockers. Patients without sinus rhythm, paced or with significant hypertensive, congenital or valvular heart disease were previously excluded. The following parameters of LV systolic function were assessed by echocardiography: fractional shortening, ejection fraction and cardiac output. LV filling was assessed by transmitral flow pulsed Doppler analysis: isovolumic relaxation time (IVRT), peak E and A wave velocities, E wave deceleration time (DT). We also evaluated a new Doppler index of combined systolic and diastolic myocardial performance (Tei index). PV (cm/s) was calculated by Color M-mode. Univariate regression analysis was performed (PV as dependent variable). RESULTS: We detected a high prevalence of diastolic function disturbances: 25 (78.1%) patients with a delayed relaxation pattern and only 3 (9.4%) with a restrictive pattern. PV did not correlate with age, body surface and LV geometry. There was a significant correlation between PV and E/A ratio (r = 0.61; p < 0.0001), IVRT (r = -0.50; p = 0.006), DT (r = -0.41; p = 0.01) and Tei index (r = -0.36; p = 0.04). CONCLUSIONS: Our results indicate that, in patients with a predominance of delayed relaxation, there is a strong relationship between PV and the classic parameters of LV filling. For similar levels of systolic impairment, PV could be an easy, fast and reproducible quantitative indicator of severity on DC.

Cardiomyopathy, Dilated↗

Receptors coupling to G proteins: is there a signal behind the sequence?

Upon the binding of their ligands, G protein-coupled receptors couple to the heterotrimeric G proteins to transduce a signal. One receptor family may couple to a single G protein subtype and another family to several ones. Is there a signal in the receptor sequence that can give an indication of the G protein subtype selectivity? We used a sequence analysis method on biogenic amine and adenosine receptors and concluded that a weak signal can be detected in receptor families where specialization for coupling to a given G protein occurred during a recent divergent evolutionary process. Proteins 2000;41:448-459.

Algorithms↗

Down-regulation of iron regulatory protein 1 gene expression by nitric oxide.

Iron regulatory protein 1 (IRP1) is an RNA binding protein that posttranscriptionally modulates the expression of mRNAs coding for proteins involved in iron metabolism. It has long been held that its RNA binding activity is regulated posttranslationally by the insertion/extrusion of a 4Fe-4S cluster, without changes in IRP1 levels. However, the question of a possible regulation of the expression of this protein has remained open. In the present study we analyzed the modulation of IRP1 expression in murine macrophages. We showed that activation by IFN-gamma and/or lipopolysaccharide, which induces IRP1 RNA binding activity via nitric oxide (NO), results simultaneously in a reduction in IRP1 protein levels, as determined by Western blot analyses. IRP1 expression decreased time-dependently to about 40% of control levels after 16 h. Down-regulation of IRP1 protein levels was correlated with the amount of NO produced and was partially abolished by the NO synthase (NOS) inhibitor N-monomethyl-l-arginine. No changes in IRP1 levels could be detected in stimulated peritoneal macrophages from NOS2 knockout (NOS2(-/-)) mice, unlike wild-type mice. Converse modulation of IRP1 RNA binding activity and IRP1 levels could be reproduced by exogenous NO and also was observed in nonmacrophage cells cocultured with NO-producing macrophages. We also analyzed IRP1 mRNA levels by Northern blotting and found a decrease in IRP1 mRNA expression after stimulation with IFN-gamma plus lipopolysaccharide, which was abrogated in the presence of N-monomethyl-l-arginine. This is evidence that IRP1 is regulated by a physiological stimulus other than posttranslationally.

Animals↗

Intracellular iron status as a hallmark of mammalian cell susceptibility to oxidative stress: a study of L5178Y mouse lymphoma cell lines differentially sensitive to H(2)O(2).

The redox properties of iron make this metal a key participant in oxygen-mediated toxicity. Accordingly, L5178Y (LY) mouse lymphoma cell lines, which display a unique inverse cross-sensitivity to ionizing radiation (IR) and hydrogen peroxide (H(2)O(2)), are a suitable model for the study of possible differences in the constitutive control of intracellular iron availability. We report here that the level of iron in the cytosolic labile iron pool (LIP), ie, potentially active in the Fenton reaction, is more than 3-fold higher in IR-resistant, H(2)O(2)-sensitive (LY-R) cells than in IR-sensitive, H(2)O(2)-resistant (LY-S) cells. This difference is associated with markedly greater content of ferritin H-subunits (H-Ft) in LY-S than in LY-R cells. Our results show that different expression of H-Ft in LY cells is a consequence of an up-regulation of H-Ft mRNA in the LY-S mutant cell line. In contrast, posttranscriptional control of iron metabolism mediated by iron-responsive element-iron regulatory proteins (IRPs) interaction is similar in the 2 cell lines, although IRP1 protein levels in iron-rich LY-R cells are twice those in iron-deficient LY-S cells. In showing that LY cell lines exhibit 2 different patterns of intracellular iron regulation, our results highlight both the role of high LIP in the establishment of pro-oxidant status in mammalian cells and the antioxidant role of ferritin. (Blood. 2000;95:2960-2966)

Aconitate Hydratase↗

Postanal repair for fecal incontinence--is it worthwhile?

PURPOSE: Patients with idiopathic or neurogenic incontinence without an isolated sphincter defect may be suitable candidates for a postanal repair. The aim of this study was to assess the results of postanal repair in patients with idiopathic or neurogenic fecal incontinence and to evaluate the role of various parameters, including preoperative physiologic testing on outcome. METHODS: Postanal repair was offered by a single surgeon to patients meeting the following criteria: incontinence score of at least 12 of 20, absence of an isolated anterior external anal sphincter defect, and failed conservative, medical, and biofeedback management. Physiologic investigation and clinical findings of female patients who had postanal repair for fecal incontinence between 1992 and 1998 were reviewed. Physiologic investigation included anorectal manometry, pudendal nerve terminal motor latency, concentric needle electromyography, and endoanal ultrasonography. Follow-up was obtained by telephone questionnaire; moreover, patients were asked to grade the outcome of their surgery as excellent or good (success) or as fair or poor (failure). RESULTS: Twenty-one patients of median age 68 (range, 40-80) years had a mean duration of fecal incontinence before postanal repair of 6.8 (range, 0.5-22) years. Twenty patients (95 percent) were available for at least one year of follow-up. Seventeen patients (80.9 percent) had at least one prior vaginal delivery, and prior sphincteroplasty had been performed in 10 patients (47.6 percent). The morbidity and mortality rates were 5 and 0 percent, respectively. After a mean follow-up period of three (range, 1-7.5) years, seven patients (35 percent) considered surgery to be successful and had a statistically significant decrease in their incontinence score. Neither prolongation of pudendal nerve terminal motor latency nor external sphincter damage as noted on electromyography or any of the preoperative manometric parameters correlated with outcome. Furthermore, patients' ages at surgery did not correlate with the degree of postoperative improvement in continence scores nor did the duration of the patients' symptoms, number of vaginal deliveries, or a history of previous surgery for fecal incontinence. CONCLUSION: None of the factors assessed was demonstrated to be predictive of outcome after postanal repair; moreover, the currently available preoperative testing has not altered the success rate, which remains low (35 percent). Despite the low success rate, the absence of any mortality and the low morbidity suggest that postanal repair may be a valid therapeutic approach. However, it should be offered only to selected patients with persistent, severe fecal incontinence despite an anatomically intact external anal sphincter who are not candidates for or refuse all other operative modalities.

Action Potentials↗