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Biomedical subjects

C Silva

Publications and source records attributed to C Silva.

At least 19 recordsLinked to original sources

Evaluation of L1 and L2 minimum norm performances on EEG localizations.

OBJECTIVE: In this work we study the performance of minimum norm methods to estimate the localization of brain electrical activity. These methods are based on the simplest forms of L(1) and L(2) norm estimates and are applied to simulated EEG data. The influence of several factors like the number of electrodes, grid density, head model, the number and depth of the sources and noise levels was taken into account. The main objective of the study is to give information about the dependence, on these factors, of the localization sources, to allow for proper interpretation of the data obtained in real EEG records. METHODS: For the tests we used simulated dipoles and compared the localizations predicted by the L(1) and L(2) norms with the location of these point-like sources. We varied each parameter separately and evaluated the results. RESULTS: From this work we conclude that, the grid should be constructed with approximately 650 points, so that the information about the orientation of the sources is preserved, especially for L(2) norm estimates; in favorable noise conditions, both L(1) and L(2) norm approaches are able to distinguish between more than one point-like sources. CONCLUSIONS: The critical dependence of the results on the noise level and source depth indicates that regularized and weighted solutions should be used. Finally, all these results are valid both for spherical and for realistic head models.

Artifacts↗

Isolation of a beta-carotene over-producing soil bacterium, Sphingomonas sp.

A carotenoid-accumulating bacterium isolated from soil, identified as a Sphingomonas sp., grew at 0.18 h(-1) and produced 1.7 mg carotenoids g(-1) dry cell, among which beta-carotene (29% of total carotenoids) and nostoxanthin (36%). A mutant strain, obtained by treatment with ethyl methanesulfonate, accumulated up to 3.5 mg carotenoids g(-1) dry cell. Accumulation of beta-carotene by this strain depended on the oxygenation of the growth medium, with maximal accumulation (89%) occurring under limiting conditions. Beta-carotene accumulation could be further enhanced by incubating the cells in the presence of glycerol (either not or only slowly assimilated) and yeast extract resulting in an accumulation of 5.7 mg beta-carotene g(-1) dry cell wt. The strain used lactose as carbon source with similar biomass and carotenoid production, providing a viable alternative use for cheese whey ultra-filtrate.

Genetic Engineering↗

Experimental investigation of dynamical coupling between turbulent transport and parallel flows in the JET plasma-boundary region.

The dynamical coupling between turbulent transport and parallel flows has been investigated in the plasma boundary region of the Joint European Torus tokamak. Experimental results show that there is a dynamical relationship between transport and parallel flows. As the size of transport events increases, parallel flows also increase. These results show that turbulent transport can drive parallel flows in the plasma boundary of fusion plasmas. This new type of measurement is an important element to unravel the overall picture connecting radial transport and flows in fusion plasmas.

Journal Article↗

Determination of lead in biological samples by use of slurry sampling electrothermal atomic absorption spectrometry.

Slurry-sampling electrothermal atomic absorption spectrometry has been applied to the determination of lead in several biological samples (fish and marine algae). The slurries were prepared both by magnetic shaking and microwave-heating and the effect of instrument operating conditions and slurry preparation conditions on the signal were examined. In addition, results from slurry sampling were compared with those obtained by microwave-assisted acid digestion of the same samples and no significant differences were found between them when the analysis of variance (ANOVA) was applied. The between-batch precision of the slurry techniques employed was similar to that for the microwave-assisted digestion procedure; values were always below 6.7%, except for the Dicentrarchus labrax sample for which the value obtained was 9.5% when using slurry magnetic shaking and 7.6% when using the slurry microwave heating. The accuracy of the slurry methodology employed was also evaluated by analysis of two biological reference materials (NIST-1577b and IAEA-V10); percentage recoveries obtained were between 95.6 and 98.5% of the values certified for lead.

Animals↗

Interchain vs. intrachain energy transfer in acceptor-capped conjugated polymers.

The energy-transfer processes taking place in conjugated polymers are investigated by means of ultrafast spectroscopy and correlated quantum-chemical calculations applied to polyindenofluorenes end-capped with a perylene derivative. Comparison between the time-integrated luminescence and transient absorption spectra measured in solution and in films allows disentangling of the contributions arising from intrachain and from interchain energy-migration phenomena. Intrachain processes dominate in solution where photoexcitation of the polyindenofluorene units induces a rather slow energy transfer to the perylene end moieties. In films, close contacts between chains favors interchain transport of the excited singlet species (from the conjugated bridge of one chain to the perylene unit of a neighboring one); this process is characterized by a 1-order-of-magnitude increase in transfer rate with respect to solution. This description is supported fully by the results of quantum-chemical calculations that go beyond the usual point-dipole model approximation and account for geometric relaxation phenomena in the excited state before energy migration. The calculations indicate a two-step mechanism for intrachain energy transfer with hopping along the conjugated chains as the rate-limiting step; the higher efficiency of the interchain transfer process is mainly due to larger electronic coupling matrix elements between closely lying chains.

Energy Transfer↗

Comparison of the genomes of two Xanthomonas pathogens with differing host specificities.

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline. Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis. Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries. Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.

Gene Order↗

Neurovirulence depends on virus input titer in brain in feline immunodeficiency virus infection: evidence for activation of innate immunity and neuronal injury.

Lentiruses cause neurological disease depending on the virus strain and its neurotropism, yet it remains uncertain to what the impact of infectious virus quantity in the brain early in infection is on the subsequent development of neurological disease or neurovirulence. We investigated the relationship between infectious virus input titer and the resulting neurovirulence, using ex vivo and in vivo assays of feline immunodeficiency virus (FIV)-induced neurovirulence. FIV infection of cell cultures and neonatal cats was performed using 10(2.5) (low-titer) or 10(4.5) (high-titer) 50% tissue culture infectious doses (TCID(50))/ml of the neurovirulent FIV strain, V1CSF. Ex vivo neurotoxicity assays revealed that conditioned medium (CM) from feline macrophages infected with high-titer (P <.001) or low-titer (P <.01) V1CSF induced greater neuronal death than CM from mock-infected cells. In vivo, animals infected intracranially with high-titer V1CSF showed neurodevelopmental delays compared to mock-infected animals (P <.001) and animals infected with low-titer V1CSF (P <.02), concurrent with reduced weight gains and greater depletion of CD4+ cells over a 12-week period. Neuropathological changes, including astrogliosis, macrophage activation, and neuronal damage, were evident in V1CSF-infected animals and were viral titer dependent. In vivo magnetic resonance (MR) spectroscopy and proton nuclear magnetic resonance ((1)H-NMR) spectroscopy of tissue extracts revealed evidence of neuronal injury, including reduced N-acetyl aspartate/creatine (P <.05) and increased trimethylamine/creatine (P <.05) ratios, in the frontal cortex of high-titer V1CSF-infected animals compared to the other groups. T2-weighted MR imaging detected increased signal intensities in the frontal cortex and white matter of V1CSF-infected animals relative to controls, which was more evident as viral titer increased (P <.01). The present findings indicate that lentivirus infectious titers in the brain during the early stages of infection determine the severity of neurovirulence, reflected by neurobehavioral deficits, together with neuroradiological and neuropathological findings of activation of innate immunity and neuronal injury.

Animals↗

Envelope gene-mediated neurovirulence in feline immunodeficiency virus infection: induction of matrix metalloproteinases and neuronal injury.

The release of neurotoxins by activated brain macrophages or microglia is one mechanism proposed to contribute to the development of neurological disease following infection by lentiviruses, including feline immunodeficiency virus (FIV). Since molecular diversity in the lentiviral envelope gene influences the expression of host molecules implicated in neuronal injury, the role of the envelope sequence in FIV neuropathogenesis was investigated by using the neurovirulent FIV strain V1CSF, the nonneurovirulent strain Petaluma, and a chimera (FIVCh) containing the V1CSF envelope gene in a Petaluma background. All three viruses replicated in primary feline macrophages with equal efficiency, but conditioned medium from V1CSF- or FIVCh-infected cells was significantly more neurotoxic than medium from Petaluma-infected cultures (P < 0.001) and could be attenuated in a dose-dependent manner by treatment with either the matrix metalloproteinase (MMP) inhibitor prinomastat (PMT) or function-blocking antibodies to MMP-2. Although FIV sequences were detectable by PCR in brain tissue from neonatal cats infected with each of the viral strains, immunohistochemistry revealed increased astrogliosis and macrophage activation in the brains of V1CSF- and FIVCh-infected cats relative to the other groups, together with elevated markers of neuronal stress that included morphological changes and increased c-fos immunoreactivity. Similarly, MMP-2, but not MMP-9, mRNA and protein expression was increased in brain tissues of V1CSF- and FIVCh-infected cats relative to Petaluma-infected animals (P < 0.01). Infection with V1CSF or FIVCh was also associated with greater CD4(+) cell depletion (P < 0.001) and neurodevelopmental delays (P < 0.005), than in Petaluma-infected animals; these deficits improved following PMT therapy. These findings indicated that diversity in the envelope gene sequence influenced the neurovirulence exhibited by FIV both in vitro and in vivo, possibly through a mechanism involving the differential induction of MMP-2.

Animals↗

[Pharmacological treatment of obesity].

The foundation on which the treatment of obesity rests is made up of the procedures aimed at unbalancing the equation of energy balance in favour of calorie consumption. Pharmacological treatment is aimed at favouring a reduction of calorie intake or stimulating calorie production. Depending on their mechanism of action, the drugs that are used in the treatment of obesity can be divided into appetite inhibitors, inhibitors of food intake or blockers of fat digestion and stimulators of thermogenesis. Amongst the appetite inhibitors, besides the adrenergic agents such as those of the amphetamine type, which are forbidden because of their addictive effect, are the serotonin uptake inhibitors such as dexfenfluramine (withdrawn in 1997) and fluoxetine. The recent commercialisation of sibutramine, a serotonin and norepinephrine reuptake inhibitor with an appetite inhibiting and thermogenesis stimulating effect, offer new perspectives in this field. Amongst inhibitors, mention is deserved by orlistat that inhibits the absorption of upto 30% of ingested fat without causing secondary effects of significance. In the area of thermogenic drugs there are the ephedrine-caffeine association, and the adrenergic beta antagonists, which are still being researched. There are numerous future perspectives amongst which are leptin, analogs of GLP-1 and cholecystokinin and neuropeptide Y antagonists. It is necessary to have reliable predictive elements that make it possible to know the most useful drugs, as well as the sort of patients who will benefit most from the different pharmacological treatments.

English Abstract↗

HIV-1 Tat neurotoxicity is prevented by matrix metalloproteinase inhibitors.

The release of potentially neurotoxic molecules by HIV-infected brain macrophages is accompanied by neuronal injury and death that results in the development of HIV-associated dementia (HAD). Among the potential neurotoxins implicated in the development of HAD is the HIV-1 transactivating protein, Tat. To investigate the mechanism by which Tat causes neurotoxicity, brain-derived Tat sequences from nondemented (Tat-ND) and demented (Tat-HAD) AIDS patients, which differed primarily in the augmenting region of Tat, were expressed in U937 monoblastoid cells and primary human macrophages. Cells expressing Tat-HAD protein exhibited elevated matrix metalloproteinase (MMP)-2 and -7 release and activation, but cells expressing Tat-ND did not exhibit enhanced MMP expression. Conditioned media from Tat-HAD-transfected cells caused significantly greater neuronal death (15.4 +/- 4.3%) than did Tat-ND (4.4 +/- 2.1%) or nontransfected (2.1 +/- 0.8%) cell-derived conditioned media. The neurotoxicity induced by Tat-HAD was inhibited by anti-MMP-2 or -7 antibodies (p < 0.005) but not by antibodies against MMP-9 or Tat. Similarly, scid/nod mice receiving striatal implants of Tat-HAD-transfected cells exhibited greater neurobehavioral abnormalities and neuronal loss (p < 0.005) than did animals receiving Tat-ND or nontransfected cells, which were reduced by treatment with the MMP inhibitor prinomastat (p < 0.005). These findings indicate that Tat causes neuronal death through an indirect mechanism that is Tat sequence dependent and involves the induction of MMPs.

AIDS Dementia Complex↗

Monocyte activation and differentiation augment human endogenous retrovirus expression: implications for inflammatory brain diseases.

Human endogenous retroviruses (HERVs) have been implicated as causative agents in diseases characterized by inflammation and macrophage activation, such as multiple sclerosis. Because monocyte activation and differentiation influence retroviral transcription and replication, we investigated the contribution of these processes to the expression of four HERV families (HERV-W, HERV-K, HERV-E, and HERV-H) in human monocytes, and autopsied brain tissue from patients with brain diseases associated with increased macrophage activity. Reverse transcriptase-polymerase chain reaction analysis of primary macrophages and U937 monocytoid cells stimulated with phorbol-12-myristate-13-acetate or lipopolysaccharide revealed three- to ninefold increases in HERV-W, HERV-K, and HERV-H RNA levels. In addition, elevated reverse transcriptase activity and HERV RNA were detectable in supernatants from PMA-stimulated U937 cultures, properties that could be attenuated with the inhibitor of monocyte differentiation threonine-lysine-proline. In contrast, stimulation of monocytes decreased or had no effect on HERV-E expression. Compared with controls, HERV-W and HERV-K expression was increased in brain tissue from patients with multiple sclerosis or human immunodeficiency virus infection or AIDS, with concomitant elevated tumor necrosis factor-alpha levels. Similarly, elevated HERV-W levels were detected in patients with Alzheimer's dementia only when tumor necrosis factor-alpha expression was also evident (2 of 6 cases). The detection of several HERVs in inflammatory brain diseases and the capacity to augment HERV expression in monocytes with compounds that influence cellular activity suggest that increased expression of these viruses is a consequence of increased immune activity rather than causative of distinct diseases.

Adult↗

Natural selection at the MJD locus: phenotypic diversity, survival and fertility among Machado-Joseph Disease patients from the Azores.

Machado-Joseph Disease (MJD) is an autosomal dominant neurodegenerative disorder of adult onset, associated with the expansion of a (CAG)n tract in the coding region of the causative gene, localized on 14q32.1. Machado-Joseph Disease shows non-Mendelian features typical of other triplet repeat disorders, including clinical heterogeneity, variable age at onset and anticipation. Three phenotypes have been proposed (clinical types 1, 2 and 3). Type 1 is associated with early age at onset and a high repeat number of the CAG sequence, and Types 2 and 3 have later onset and lower numbers of CAG repeats. This paper investigates whether there is selection against the MJD gene, acting through differential survival. nuptiality and fertility associated with clinical type and age at onset. The study sample comprised 40 MJD patients from the Azores (Portugal) having fully documented reproductive histories and known dates of death. The proportion of married patients of each clinical type increased from 0.22 among Type 1 patients, to 0.40 in Type 2 and 0.95 in Type 3. Age at onset and length of survival were also associated with marital status, with the married cases having later mean age at onset and longer mean survival time. In the whole sample, clinical type was associated with fertility, with significantly fewer children born to Type 1 patients. Among married patients clinical type was not associated with age at marriage, reproductive span or number of children. No reduction of fertility was detected among married patients in whom the onset of MJD was below the age of 50. The authors' interpretation of these results is that the high-repeat CAG haplotypes associated with early age at onset and clinical Type 1 are selected against through reduced survival and fertility. The fertility component of selection is mediated by nuptiality rather than marital fertility.

Adolescent↗

Encephalopathy with staphylococcal endocarditis: multiple neuropathological findings.

BACKGROUND: Infective endocarditis is associated with serious neurological sequelae. OBJECTIVE: Here, we report a patient with Staphylococcus aureus endocarditis, secondary to congenital heart disease, with subacute onset of multiple neurological complications. RESULTS: Despite prompt antibiotic treatment with rapid sterilization of blood cultures, the patient died with brain herniation within 96 hours of admission. Neuropathological examination showed intraparenchymal hemorrhages, mycotic aneurysms, micro-abscesses and septic arteritis with accompanying infarction. Immunocytochemical studies revealed enhanced CD45 and GFAP immunoreactivity, together with adenosine A1 receptor detection on macrophages and microglia. CONCLUSIONS: Infective endocarditis is associated with multiple neuropathological lesions, which may contribute to its poor clinical outcome and activation of cells of monocyte-microglial lineage throughout the brain.

Adult↗

Clinical and genetic heterogeneity in hereditary haemochromatosis: association between lymphocyte counts and expression of iron overload.

To identify a new marker of expression of disease, independent of HFE genotype in patients with hereditary haemochromatosis (HHC), the total peripheral blood lymphocyte counts were analysed according to iron status in two groups of subjects with HFE mutations. The groups consisted of 38 homozygotes for C282Y, and 107 heterozygotes for the C282Y or compound heterozygotes for C282Y and H63D. For control purposes, total lymphocyte counts and iron status were also examined in 20 index patients with African dietary iron overload, a condition not associated with HFE mutations, and in 144 members of their families and communities. Mean lymphocyte numbers were lower in C282Y homozygous HHC index subjects with cirrhosis and higher iron stores than in those without cirrhosis and with lower iron burdens [(1.65 +/- 0.43) x 10(6)/mL vs. (2.27 +/- 0.49) x 10(6)/mL; p = 0.008]. Similarly, mean lymphocyte counts were significantly lower in C282Y heterozygotes and C282Y/H63D compound heterozygotes with iron overload and increased serum ferritin concentrations compared to those with normal serum ferritin concentrations (p < 0.05). Statistically significant negative correlations were found, in males, between lymphocyte counts and the total body iron stores, either in C282Y homozygous HHC patients (p = 0.031 in a multiple regression model dependent on age) and in C282Y heterozygotes or C282Y/H63D compound heterozygotes with iron overload (p = 0.029 in a simple linear model). In contrast, lymphocyte counts increased with increasing serum ferritin concentrations among the index subjects with African iron overload (r = 0.324, not statistically significant) and among the members of their families and communities (r = 0.170, p = 0.042). These results suggest that a lower peripheral blood lymphocyte count is associated with a greater degree of iron loading in HFE haemochromatosis but not in African iron overload, and they support the notion that the lymphocyte count may serve as a marker of a non-HFE gene that influences the clinical expression of HFE haemochromatosis.

Adult↗

A light- and electron-microscopic histopathological study of human bladder mucosa after intravesical resiniferatoxin application.

OBJECTIVE: To determine the morphology of bladder mucosa and the integrity of its mucin coat in patients with detrusor hyper-reflexia treated with intravesical resiniferatoxin. PATIENTS AND METHODS: Seven patients with detrusor hyper-reflexia were treated intravesically with resiniferatoxin dissolved in 10% ethanol in saline (50 nmol/L solution in two and 100 nmol/L in five). Patients were clinically evaluated by a voiding chart and filling cystometry before and 3 months after each resiniferatoxin application. In addition, they underwent cystoscopy and bladder biopsies at 22-33 months after the first instillation and at 7-23 months after the last one. Tissue samples for light microscopy were fixed in 4% paraformaldehyde, embedded in paraffin and stained with haematoxylin-eosin or periodic acid-Schiff reagent (PAS). Those for electron microscopy were fixed in 5% glutaraldehyde and embedded in resin. RESULTS: The resiniferatoxin instillation was not painful. Three months after treatment the mean voiding frequency decreased and five incontinent patients became continent. The maximum cystometric capacity increased in all patients; at cystoscopy the bladders appeared normal. On light microscopy the urothelium was of normal morphology and stained with PAS in the luminal cells and in the basement membrane. Mononuclear inflammatory cells were occasionally apparent in the lamina propria. On electron microscopy epithelial cells were visible in a thick basal lamina. Superficial cells had the usual irregular contour and contained numerous membrane-coated vesicles. In the lamina propria, unmyelinated axonal profiles with occasional varicosities could be identified. CONCLUSIONS: Intravesical resiniferatoxin improved urinary frequency, incontinence and bladder capacity in patients with detrusor hyper-reflexia, causing no morphological change in the bladder mucosa. The PAS reactivity of the carbohydrate moieties present in the mucin coat and the basement membrane was unchanged by resiniferatoxin.

Administration, Intravesical↗

T-cell reactivity against streptococcal antigens in the periphery mirrors reactivity of heart-infiltrating T lymphocytes in rheumatic heart disease patients.

T-cell molecular mimicry between streptococcal and heart proteins has been proposed as the triggering factor leading to autoimmunity in rheumatic heart disease (RHD). We searched for immunodominant T-cell M5 epitopes among RHD patients with defined clinical outcomes and compared the T-cell reactivities of peripheral blood and intralesional T cells from patients with severe RHD. The role of HLA class II molecules in the presentation of M5 peptides was also evaluated. We studied the T-cell reactivity against M5 peptides and heart proteins on peripheral blood mononuclear cells (PBMC) from 74 RHD patients grouped according to the severity of disease, along with intralesional and peripheral T-cell clones from RHD patients. Peptides encompassing residues 1 to 25, 81 to 103, 125 to 139, and 163 to 177 were more frequently recognized by PBMC from RHD patients than by those from controls. The M5 peptide encompassing residues 81 to 96 [M5(81-96) peptide] was most frequently recognized by PBMC from HLA-DR7+ DR53+ patients with severe RHD, and 46.9% (15 of 32) and 43% (3 of 7) of heart-infiltrating and PBMC-derived peptide-reactive T-cell clones, respectively, recognized the M5(81-103) region. Heart proteins were recognized more frequently by PBMC from patients with severe RHD than by those from patients with mild RHD. The similar pattern of T-cell reactivity found with both peripheral blood and heart-infiltrating T cells is consistent with the migration of M-protein-sensitized T cells to the heart tissue. Conversely, the presence of heart-reactive T cells in the PBMC of patients with severe RHD also suggests a spillover of sensitized T cells from the heart lesion.

Antigen Presentation↗

Progesterone does not alter osmotic regulation of AVP.

To test the hypothesis that progesterone, independent of estrogen, decreases the plasma osmotic threshold for arginine vasopressin (AVP) release and thirst onset, we compared AVP and thirst responses to hypertonic saline infusion (HSI) during administration of oral contraceptives (OCs) containing progesterone (OCP) with responses to infusion of OCs containing progesterone and estrogen (OCEP). Eight women (29 +/- 2 yr) were infused with 3% NaCl (120 min, 0.1 ml. kg body wt(-1). min(-1)) and consumed fluid (90 min, 15 ml/kg body wt) in the early follicular and midluteal phases of a 28-day menstrual cycle and also after 4 wk of OCP and after 4 wk of OCEP in a randomized crossover design. Baseline plasma osmolality (P(osm)) was lower in the luteal phase (280 +/- 1 mosmol/kgH(2)O) and during OCEP (283 +/- 1 mosmol/kgH(2)O) than in the follicular phase (286 +/- 1 mosmol/kgH(2)O, P < 0.05) but was unaffected by OCP (284 +/- 1 mosmol/kgH(2)O). P(osm) remained lower in the follicular phase than in the luteal phase and with OCEP throughout the first 50 min of HSI. The mean abscissal plasma AVP concentration-P(osm) intercept was unaffected by OCP (267 +/- 1 mosmol/kgH(2)O) but was greater in the follicular phase (273 +/- 2 mosmol/kgH(2)O) than in the luteal phase (266 +/- 4 mosmol/kgH(2)O) and with OCEP (268 +/- 2 mosmol/kgH(2)O, P < 0.05). There were no differences in osmotic thresholds for thirst onset across experimental days. Despite the lower osmotic threshold for AVP release during the luteal phase and with OCEP, fluid balance, renal free water clearance, and Na(+) regulation during HSI were unaffected by menstrual phase or OC treatment, indicating a lower osmotic operating point for body water balance. OCP did not affect osmotic AVP regulation, suggesting that progesterone does not affect osmotic fluid regulation through a mechanism independent of estrogen.

Adult↗