Biomedical subjects

G Pereira

Publications and source records attributed to G Pereira.

At least 19 recordsLinked to original sources

PubMed2007

Metal-chelating compounds produced by ectomycorrhizal fungi collected from pine plantations.

AIMS: To investigate the in vitro production of metal-chelating compounds by ectomycorrhizal fungi collected from pine plantations in southern Chile. METHODS AND RESULTS: Scleroderma verrucosum, Suillus luteus and two isolates of Rhizopogon luteolus were grown in solid and liquid modified Melin-Norkans (MMN) media with and without iron addition and the production of iron-chelating compounds was determined by Chrome Azurol S (CAS) assay. The presence of hydroxamate and catecholate-type compounds and organic acids was also investigated in liquid medium. All isolates produced iron-chelating compounds as detected by CAS assay, and catecholates, hydroxamates as well as oxalic, citric and succinic acids were also detected in all fungal cultures. Scleroderma verrucosum produced the greatest amounts of catecholates and hydroxamates whereas the highest amounts of organic acids were detected in S. luteus. Nevertheless, the highest catecholate, hydroxamate and organic acid concentrations did not correlate with the highest CAS reaction which was observed in R. luteolus (Yum isolate). CONCLUSIONS: Ectomycorrhizal fungi produced a variety of metal-chelating compounds when grown in liquid MMN medium. However, the addition of iron to all fungi cultures reduced the CAS reaction, hydroxamate and organic acid concentrations. Catecholate production was affected differently by iron, depending on the fungal isolate. SIGNIFICANCE AND IMPACT OF THE STUDY: The ectomycorrhizal fungi described in this study have never been reported to produce metal-chelating compound production. Moreover, apart from some wood-rotting fungi, this is the first evidence of the presence of catecholates in R. luteolus, S. luteus and S. verrucosum cultures.

Basidiomycota↗

PubMed2006

Presence of Shiga toxin-producing E. coli O157:H7 in a survey of wild artiodactyls.

This study was carried out to evaluate the role of wild artiodactyls as reservoirs of Escherichia coli O157:H7 for livestock and humans. Retroanal mucosal swabs samples from 206 red deer (Cervus elaphus), 20 roe deer (Capreolus capreolus), 6 fallow deer (Dama dama) and 11 mouflon (Ovis musimon), collected during the hunting season (autumn-winter) in South-western Spain, were screened. Samples were pre-enriched in modified buffered peptone water, concentrated by an immunomagnetic separation technique and cultured onto selective cefixime tellurite sorbitol MacConkey agar. Polymerase chain reaction (PCR) was used to detect the presence of genes coding O157 and H7 antigens and the virulence factors verocytotoxin, intimin and enterohaemolysin. Three E. coli O157:H7 isolates were obtained from red deer (1.5%). Two of them showed inability to ferment sorbitol and lack of beta-d-glucuronidase (GUD) activity, however, the other strain investigated was an atypical sorbitol-fermenting E. coli O157:H7 with GUD(+) activity. This is the first report pointing to red deer as a reservoir of E. coli O157:H7 in Spain.

Adhesins, Bacterial↗

PubMed2005

Bovine tuberculosis in wild boar (Sus scrofa), red deer (Cervus elaphus) and cattle (Bos taurus) in a Mediterranean ecosystem (1992-2004).

During the last 12 years, an increasing frequency in condemnation of hunted red deer and wild boar carcasses due to the presence of tubercle-like lesions has been observed in Extremadura (Western Spain). Before 1993, tuberculosis was a very rare finding in hunted animals. The current tuberculosis regional prevalence in cattle approaches 0.4% after years of expensive test and slaughter campaigns. It is imperative to investigate the epidemiology of Mycobacterium bovis infection in red deer and wild boar in order to keep a good health status and to maintain the effectiveness of domestic species TB eradication programs. The present paper evaluates the problem in Sierra de San Pedro, estimating the prevalence of TB in wild boar and red deer, the main wild artiodactyls in the area, and domestic cattle since 1992-2004, by the use of a low-cost surveillance method based on detailed pathological inspection of hunted animal carcasses. Microbiology and molecular epidemiology studies on several M. bovis isolates from domestic and wild animals helped to define the interspecies contacts. These findings, as well as recent history of game estates management and descriptive epidemiology field work, throw light on the rise and maintenance of these epizootics.

Animals↗

PubMed2001

Modes of spindle pole body inheritance and segregation of the Bfa1p-Bub2p checkpoint protein complex.

Yeast spindle pole bodies (SPBs) duplicate once per cell cycle by a conservative mechanism resulting in a pre-existing 'old' and a newly formed SPB. The two SPBs of yeast cells are functionally distinct. It is only the SPB that migrates into the daughter cell, the bud, which carries the Bfa1p-Bub2p GTPase-activating protein (GAP) complex, a component of the spindle positioning checkpoint. We investigated whether the functional difference of the two SPBs correlates with the time of their assembly. We describe that in unperturbed cells the 'old' SPB always migrates into the bud. However, Bfa1p localization is not determined by SPB inheritance. It is the differential interaction of cytoplasmic microtubules with the mother and bud cortex that directs the Bfa1p-Bub2p GAP to the bud-ward-localized SPB. In response to defects of cytoplasmic microtubules to interact with the cell cortex, the Bfa1p-Bub2p complex binds to both SPBs. This may provide a mechanism to delay cell cycle progression when cytoplasmic microtubules fail to orient the spindle. Thus, SPBs are able to sense cytoplasmic microtubule properties and regulate the Bfa1p-Bub2p GAP accordingly.

Antineoplastic Agents↗

PubMed2001

The role of the yeast spindle pole body and the mammalian centrosome in regulating late mitotic events.

Centrosomes of vertebrate cells and spindle pole bodies (SPBs) of fungi were first recognized through their ability to organize microtubules. Recent studies suggest that centrosomes and SPBs also have a function in the regulation of cell cycle progression, in particular in controlling late mitotic events. Regulators of mitotic exit and cytokinesis are associated with the SPB of budding and fission yeast. Elucidation of the molecular roles played by these regulators is helping to clarify the function of the SPB in controlling progression though mitosis.

Animals↗

PubMed2001

Microbial respiration and chemical composition of different sediment fractions in waterbodies of the upper Paraná River floodplain, Brazil.

Four size fractions of the sediment of six environments of the upper Paraná River floodplain were analyzed for carbon, nitrogen, and phosphorus contents and microbial respiration (oxygen consumption). Particle size did not affect nitrogen and phosphorus content or microbial activity, but did affect carbon content (F = 4.274, df = 3; 20, p = 0.020). The carbon concentration of ultra-fine particles was significantly lower than that of other sizes of sediment particles. Microbial respiration values were well predicted by sediment chemical composition, as shown by multiple regression (microbial respiration = -0.39 - 0.210C + 0.108N + 0.796P; F = 7.0495, p = 0.0022). However, phosphorus was the element which best explained the microbial respiration (partial coefficient = 0.796, p = 0.0039, n = 23). Considering that i) phosphorus was the best predictor of microbial respiration; ii) phosphorus is trapped in the series of reservoirs located upstream from the section of the floodplain studied; and iii) microbial respiration is a measure of decomposition rates and nutrient cycling, we hypothesize that the long-term accumulation of litter detritus and reduction of nutrient cycling in environments of the upper Paraná River floodplain are probable impacts of this decrease in phosphorus caused by the upstream reservoirs.

Bacteria↗

PubMed2001

[Characteristics of falls among free living elders].

BACKGROUND: Falls by older adults are a frequent problem among ambulatory patients in primary care. AIM: To describe the prevalence and features of falls among elders consulting to an ambulatory Geriatric Clinic. PATIENTS AND METHODS: Persons aged 60 years or more were surveyed about the number of falls in the preceding six months, the characteristics and consequences of each falls. Biopsychosocial characteristics were recorded and the Tinetti gait and balance test was performed in all patients reporting falls. RESULTS: In 104 (18.2%) of 571 clinical consultations, one or more falls were reported. Among patients who fell and provided complete data (n = 95), 64% reported one fall and 36% reported two or more, totaling 156 falls to analyze. The mean age of the patients with falls was 71.8 +/- 6.8 years. The functional and cognitive status was normal in 73 and 71.6% of cases respectively and 38% carried out periodical physical activity. Fifty seven percent of falls occurred outside of home, and an extrinsic factor was a precipitating cause in 55% of the falls. A post-fall syndrome appeared in 21% of cases and 2.6% resulted in fractures. Falling two or more times versus one time during the last six months was statistically associated with an age over 75, an absence of periodic physical activity, functional impairments, three or more chronic diseases, neurological diseases and with living alone, among other variables. CONCLUSIONS: Falls among elders occur mainly outside of home, in subjects older than 75 years old, functionally dependent and with an important involvement of extrinsic factors. Physical activity, as well as the control of environmental risks, could be protective factors against recurrent falls.

Accidental Falls↗

PubMed2000

Distribution, targeting, and internalization of the sst4 somatostatin receptor in rat brain.

Somatostatin mediates its diverse physiological effects through a family of five G-protein-coupled receptors (sst(1)-sst(5)); however, knowledge about the distribution of individual somatostatin receptor proteins in mammalian brain is incomplete. In the present study, we have examined the regional and subcellular distribution of the somatostatin receptor sst(4) in the rat CNS by raising anti-peptide antisera to the C-terminal tail of sst(4). The specificity of affinity-purified antibodies was demonstrated using immunofluorescent staining of HEK 293 cells stably transfected with an epitope-tagged sst(4) receptor. In Western blotting, the antiserum reacted specifically with a broad band in rat brain, which migrated at approximately 70 kDa before and approximately 50 kDa after enzymatic deglycosylation. sst(4)-Like immunoreactivity was most prominent in many forebrain regions, including the cerebral cortex, hippocampus, striatum, amygdala, and hypothalamus. Analysis at the electron microscopic level revealed that sst(4)-expressing neurons target this receptor preferentially to their somatodendritic domain. Like the sst(2A) receptor, sst(4)-immunoreactive dendrites were often closely apposed by somatostatin-14-containing fibers and terminals. However, unlike the sst(2A) receptor, sst(4) was not internalized in response to intracerebroventricular administration of somatostatin-14. After percussion trauma of the cortex, neuronal sst(4) receptors progressively declined at the sites of damage. This decline coincided with an induction of sst(4) expression in cells with a glial-like morphology. Together, this study provides the first description of the distribution of immunoreactive sst(4) receptor proteins in rat brain. We show that sst(4) is strictly somatodendritic and most likely functions in a postsynaptic manner. In addition, the sst(4) receptor may have a previously unappreciated function during the neuronal degeneration-regeneration process.

Age Factors↗

PubMed2000

The Bub2p spindle checkpoint links nuclear migration with mitotic exit.

Bfa1p and Bub2p are spindle checkpoint proteins that likely have GTPase activation activity and are associated with the budding yeast spindle pole body (SPB). Here, we show that Bfa1p and Bub2p bind the Ras-like GTPase Tem1p, a component of the mitotic exit network, to the cytoplasmic face of the SPB that enters the bud, whereas the GDP/GTP exchange factor Lte1p is associated with the cortex of the bud. Migration of the SPB into the bud probably allows activation of Tem1p through Lte1p, thereby linking nuclear migration with mitotic exit. Since components of the Bub2p checkpoint are conserved in other organisms, we propose that the position of the SPB or mammalian centrosome controls the timing of mitotic exit.

Cell Cycle↗

PubMed2000

[Eclossion of cysts of two species of Dendrocephalus (Anostraca: Thamnocephalidae) of potential use as food in aquaculture].

The use of Artemia salina nauplii as live food has significantly aided culture of commercial fish and shrimps in recent years. However, reported deficiencies in the nutritional value of some strains originated the study of freshwater fairy shrimps as an alternative to Artemia. This study presents cyst biometry for Dendrocephalus geayi and D. spartaenovae (Anostraca: Thamnocephalidae), and the effects of some physicochemical variables on cyst hatching. The biometric characteristics of D. geayi and D. spartaenovae are within the size range of commercial Artemia strains. Favorable conditions to hatch D. geayi are 30 degrees C and conductivity near that of distilled water (<5 micromhos). For D. spartaenovae, these conditions are 28 degrees C and 280 micromhos. A very low salinity such as 1 per thousand inhibits hatching in both species.

Animal Feed↗

PubMed1999

Interaction of the yeast gamma-tubulin complex-binding protein Spc72p with Kar1p is essential for microtubule function during karyogamy.

The spindle pole body component Kar1p has a function in nuclear fusion during conjugation, a process known as karyogamy. The molecular role of Kar1p during this process is poorly understood. Here we show that the yeast gamma-tubulin complex-binding protein Spc72p interacts directly with the N-terminal domain of Kar1p, thereby targeting the gamma-tubulin complex to the half bridge, a substructure of the spindle pole body, where it organizes microtubules. This binding of Spc72p to Kar1p has only a minor role during vegetative growth, whereas it becomes essential for karyogamy in mating cells, explaining the important role of Kar1p in this process. We also show that the localization of Spc72p within the spindle pole body changes throughout the cell cycle and even more strongly in response to mating pheromone. Taken together, these observations suggest that the relocalization of Spc72p within the spindle pole body is the 'landmark' event in the pheromone-induced reorganization of the cytoplasmic microtubules.

Fungal Proteins↗

PubMed1999

Epitope tagging of yeast genes using a PCR-based strategy: more tags and improved practical routines.

Epitope tagging of proteins as a strategy for the analysis of function, interactions and the subcellular distribution of proteins has become widely used. In the yeast Saccharomyces cerevisiae, molecular biological techniques have been developed that use a simple PCR-based strategy to introduce epitope tags to chromosomal loci (Wach et al., 1994). To further employ the power of this strategy, a variety of novel tags was constructed. These tags were combined with different selectable marker genes, resulting in PCR amplificable modules. Only one set of primers is required for the amplification of any module. Furthermore, convenient laboratory techniques are described that facilitate the genetic manipulations of yeast strains, as well as the analysis of the epitope-tagged proteins.

Blotting, Western↗

PubMed1999

Intercomparison of calibration procedures for 192Ir HDR sources in Brazil.

The lack of well established dosimetry protocols for HDR sources is a point of great concern regarding the uniformity of procedures within a particular country and worldwide. The main objective of this paper is to report the results from ten institutions of an intercomparison of calibration procedures for 192Ir HDR sources currently in use in Brazil. The treatment irradiator of one institution was calibrated by a reference system and used by all participants with their own measuring electrometers and ionization chambers under the same experimental conditions. Two methods were used: the calibration jig and the well-type ionization chamber. Each participant was allowed to use their own method and formalism. The results of this exercise were very positive since this was the first time in Brazil that a group of users gathered to share their experience and openly discuss the physical concepts behind the calibration procedures. The results were all within +/-3.0%, except one case where -4.6% was observed and later identified as a problem with the Nk value for x-rays. Though the magnitude of the deviations found was generally acceptable considering the diversity of formalisms currently in use, a proposal is now being prepared to be adopted as a national protocol. The identification of the institutions was left out for the sake of confidentiality.

Brachytherapy↗

PubMed1999

Evaluation of a long-chain polyunsaturated fatty acid supplemented formula on growth, tolerance, and plasma lipids in preterm infants up to 48 weeks postconceptional age.

BACKGROUND: The last trimester of pregnancy is a period of rapid accretion of long-chain polyunsaturated fatty acids, both in the central nervous system and the body as a whole. Human milk contains these fatty acids, whereas some preterm infant formulas do not. Infants fed formulas without these fatty acids have lower plasma and erythrocyte concentrations than infants fed human milk. Preclinical and clinical studies have demonstrated that single-cell sources (algal and fungal) of long-chain polyunsaturated fatty acids are bioavailable. A balanced addition of fatty acids from these oils to preterm formula results in blood fatty acid concentrations in low birth weight infants comparable to those of infants fed human milk. METHODS: In the present study the growth, acceptance (overall incidence of discontinuation, reasons for discontinuation, overall incidence and type of individual adverse events), and plasma fatty acid concentrations were compared in three groups of infants fed a long-chain polyunsaturated fatty acid-supplemented preterm infant formula, an unsupplemented control formula, or human milk. The study was prospective, double-blind (formula groups only), and randomized (formula groups only). Two hundred eighty-eight infants were enrolled (supplemented formula group, n = 77; control formula group, n = 78; human milk group, n = 133). RESULTS: Anthropometric measurements at enrollment, at first day of full oral feeding, and at both 40 and 48 weeks postconceptional age did not differ between the formula groups, whereas the human milk-fed group initially grew at a lower rate. The incidence of severe adverse events was rare and not significantly different between formula groups. The groups fed either human milk or supplemented formula had long-chain polyunsaturated fatty acid concentrations higher than those in the control formula group. CONCLUSIONS: The results of this study demonstrate the safety and efficacy of a preterm formula supplemented with long-chain polyunsaturated fatty acids from single-cell oils.

Aging↗

PubMed1999

[The implantation of VDD pacemakers with a single electrode--a comparative study. The experience of the last 5 years].

OBJECTIVE: To compare the epidemiological characteristics and immediate results of all first single lead VDD pacemaker (PM) implantations with those of an equal number of dual chamber DDD PM, implanted during a 5-year period in a tertiary-care hospital. POPULATION AND METHODS: A total of 41 patients (pts) (25 males, mean age of 69.0 +/- 11.8 years) underwent a VDD PM implantation, from 30-11-92 to 15-9-97. This group was compared with an equal number of patients (28 males, mean age of 69.9 +/- 7.31 years) with a DDD PM implanted in the same period, selected by a criterion of immediate temporal proximity of procedure. For each patient we collected the clinical and electrocardiographic (ECG) indications for PM implantation, parameters of atrial (AS) and ventricular (VS) sensing and ventricular pacing (VP), X-ray exposure time (XRT) and complications. RESULTS: In the VDD group, 46.3% of the patients had syncope, 51.2% had complete AV block on the ECG, and 14.6% were PM-dependent. Analyzed procedure-related parameters were as follows: P-wave amplitude: 2.1 +/- 0.6 V; AS threshold: 1.2 +/- 0.7 V; R-wave amplitude: 9.1 +/- 3.3 V; VS threshold: 7.0 +/- 2.0 V; VP thresholds: 0.68 +/- 0.24 mA, 0.43 +/- 0.12 V (for a spike duration of 0.5 ms); ventricular impedance: 644.9 +/- 132.0 ohm; XRT; 7' 43" +/- 8' 23". There were two minor complications, for an incidence of 4.9% (one local hematoma and a vagal reaction). In the DDD group the clinical and ECG characteristics were similar, but there was a 22.0% prevalence of sinus-node dysfunction, VS 0% in the VDD group). The P-wave amplitude and AS threshold were significantly (p < 0.005) better (2.8 +/- 0.9 V and 2.8 +/- 0.9 V respectively). The other parameters were similar to those of the VDD group. CONCLUSIONS: The immediate results of VDD PM implantation are good and comparable with those of DDD PM, although with worse acute AS parameters.

Aged↗

PubMed1998

Spc98p directs the yeast gamma-tubulin complex into the nucleus and is subject to cell cycle-dependent phosphorylation on the nuclear side of the spindle pole body.

In the yeast Saccharomyces cerevisiae, microtubules are organized by the spindle pole body (SPB), which is embedded in the nuclear envelope. Microtubule organization requires the gamma-tubulin complex containing the gamma-tubulin Tub4p, Spc98p, and Spc97p. The Tub4p complex is associated with cytoplasmic and nuclear substructures of the SPB, which organize the cytoplasmic and nuclear microtubules. Here we present evidence that the Tub4p complex assembles in the cytoplasm and then either binds to the cytoplasmic side of the SPB or is imported into the nucleus followed by binding to the nuclear side of the SPB. Nuclear import of the Tub4p complex is mediated by the essential nuclear localization sequence of Spc98p. Our studies also indicate that Spc98p in the Tub4p complex is phosphorylated at the nuclear, but not at the cytoplasmic, side of the SPB. This phosphorylation is cell cycle dependent and occurs after SPB duplication and nucleation of microtubules by the new SPB and therefore may have a role in mitotic spindle function. In addition, activation of the mitotic checkpoint stimulates Spc98p phosphorylation. The kinase Mps1p, which functions in SPB duplication and mitotic checkpoint control, seems to be involved in Spc98p phosphorylation. Our results also suggest that the nuclear and cytoplasmic Tub4p complexes are regulated differently.

Amino Acid Sequence↗

PubMed1997

The spindle pole body component Spc97p interacts with the gamma-tubulin of Saccharomyces cerevisiae and functions in microtubule organization and spindle pole body duplication.

Previously, we have shown that the gamma-tubulin Tub4p and the spindle pole body component Spc98p are involved in microtubule organization by the yeast microtubule organizing centre, the spindle pole body (SPB). In this paper we report the identification of SPC97 encoding an essential SPB component that is in association with the SPB substructures that organize the cytoplasmic and nuclear microtubules. Evidence is provided for a physical and functional interaction between Tub4p, Spc98p and Spc97p: first, temperature-sensitive spc97(ts) mutants are suppressed by high gene dosage of SPC98 or TUB4. Second, Spc97p interacts with Spc98p and Tub4p in the two-hybrid system. Finally, immunoprecipitation and fractionation studies revealed complexes containing Tub4p, Spc98p and Spc97p. Further support for a direct interaction of Tub4p, Spc98p and Spc97p comes from the toxicity of strong SPC97 overexpression which is suppressed by co-overexpression of TUB4 or SPC98. Analysis of temperature-sensitive spc97(ts) alleles revealed multiple spindle defects. While spc97-14 cells are either impaired in SPB separation or mitotic spindle formation, spc97-20 cells show an additional defect in SPB duplication. We discuss a model in which the Tub4p-Spc98p-Spc97p complex is part of the microtubule attachment site at the SPB.

Cloning, Molecular↗

PubMed1997

Centrosome-microtubule nucleation.

In many cell types the formation of microtubules from tubulin subunits is initiated at defined nucleation sites at the centrosome. These sites contain the conserved gamma-tubulin which is in association with additional not very will characterised proteins, identified as components of a gamma-tubulin ring complex from Xenopus egg extracts or from suppressor screens in the yeast Saccharomyces cerevisiae. In this review we discuss two recently proposed models of how the gamma-tubulin complex assists in the assembly of tubulin to form microtubules. These models propose different roles for gamma-tubulin and the other proteins in the complex in tubulin assembly. While the structure and composition of a microtubule nucleation site is becoming clearer, it is still unknown how the cell-cycle dependent regulation of microtubule nucleation sites is achieved and whether they disassemble after microtubule formation in order to allow microtubule fluxes towards the centrosome which have been observed in mitotic cells.

Animals↗