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Biomedical subjects

M E Pereira

Publications and source records attributed to M E Pereira.

At least 19 recordsLinked to original sources

Mercury in urban soils: a comparison of local spatial variability in six European cities.

The objective of this study was to quantify and assess for the first time the variability of total mercury in urban soils at a European level, using a systematic sampling strategy and a common methodology. We report results from a comparison between soil samples from Aveiro (Portugal), Glasgow (Scotland), Ljubljana (Slovenia), Sevilla (Spain), Torino (Italy) and Uppsala (Sweden). At least 25 sampling points (in about 4-5 ha) from a park in each city were sampled at two depths (0-10 and 10-20 cm). Total mercury was determined by pyrolysis atomic absorption spectrometry with gold amalgamation. The quality of results was monitored using certified reference materials (BCR 142R and BCR 141R). Measured total mercury contents varied from 0.015 to 6.3 mg kg(-1). The lowest median values were found in Aveiro, for both surface (0-10 cm) and sub-surface (10-20 cm) samples (0.055 and 0.054 mg kg(-1), respectively). The highest median mercury contents in soil samples were found in samples from Glasgow (1.2 and 1.3 mg kg(-1), for surface and sub-surface samples, respectively). High variability of mercury concentrations was observed, both within each park and between cities. This variability reflecting contributions from natural background, previous anthropogenic activities and differences in the ages of cities and land use, local environmental conditions as well as the influence of their location within the urban area. Short-range variability of mercury concentrations was found to be up to an order of magnitude over the distance of only a few 10 m.

Cities↗

Effectiveness of ZnCl2 in protecting against nephrotoxicity induced by HgCl2 in newborn rats.

This work investigated the preventive effects of ZnCl(2) on renal and hepatic alterations induced by HgCl(2) in young rats. Wistar rats of 3 days old were treated (s.c.) on consecutive days with saline or ZnCl(2) 27 mg/kg/day from the 3rd to the 7th and with saline or HgCl(2) 5.0mg/kg/day from the 8th to the 12th day of life. Pups were sacrificed 24h after the last dose and samples were collected. The creatinine and urea dosages, used as renal parameters, presented increases of 35% and 500%, respectively. The alanine aminotransferase and lactic dehydrogenase activities, used as hepatic parameters, presented a decrease (40%) and no alteration, respectively, by mercury exposure. The glycemia was diminished and the hepatic glycogen was not modified by mercury. All the mercury effects were prevented by zinc. These results suggest that mercury intoxication of young rats alters the renal function but does not modify the hepatic parameters, and previous exposure to zinc is able to avoid the renal damage.

Alanine Transaminase↗

Can Nassarius reticulatus be used as a bioindicator for Hg contamination? Results from a longitudinal study of the Portuguese coastline.

The focus of this work is to explore the use of the netted whelk, Nassarius reticulatus (L.), as an indicator of mercury (Hg) contamination, by assessing the concentration of Hg in the sediments and in the whelk along the entire Portuguese coast. Total Hg concentrations ranged from below the detection limit (0.01 ng absolute mercury) up to 0.87 mg kg(-1) dry weight (dwt) in sediments and between 0.06 and 1.02 mg kg(-1) (dwt) for organisms, with no significant differences between males and females. Although organic mercury was not detected in the sediments, it represented, on average, 52% of the total Hg in the whelk tissues, and as high as 88% in some cases, suggesting mercury accumulation from dietary intake. Significant negative correlations were found between the total Hg concentrations in the sediments and the log(10) of Hg concentrations in whelk tissues males (r=-0.64; P<0.01) and females (r=-0.52; P<0.01) indicating that the species is a poor indicator of Hg contamination. Nevertheless, since the highest concentrations of organic mercury in the whelk tissues were found in the least contaminated areas, this species must be highly relevant in the trophic web, namely on the possible biomagnification of mercury. The high dietary mercury accumulation from feeding on carrion and the low bioavailability of mercury to whelks in estuarine sediments may be the basis of the mercury accumulation pattern in N. reticulatus.

Animals↗

Perspectives of digestive pest control with proteinase inhibitors that mainly affect the trypsin-like activity of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae).

The present study describes the main characteristics of the proteolytic activities of the velvetbean caterpillar, Anticarsia gemmatalis Hübner, and their sensitivity to proteinase inhibitors and activators. Midguts of last instar larvae reared on an artificial diet were homogenized in 0.15 M NaCl and centrifuged at 14,000 g for 10 min at 4 degrees C and the supernatants were used in enzymatic assays at 30 degrees C, pH 10.0. Basal total proteolytic activity (azocasein hydrolysis) was 1.14 +/- 0.15 absorbance variation min(-1) mg protein(-1), at 420 nm; basal trypsin-like activity (N-benzoyl-L-arginine-p-nitroanilide, BApNA, hydrolysis) was 0.217 +/- 0.02 mmol p-nitroaniline min(-1) mg protein(-1). The maximum proteolytic activities were observed at pH 10.5 using azocasein and at pH 10.0 using BApNA, this pH being identical to the midgut pH of 10.0. The maximum trypsin-like activity occurred at 50 degrees C, a temperature that reduces enzyme stability to 80 and 60% of the original, when pre-incubated for 5 and 30 min, respectively. Phenylmethylsulfonyl fluoride inhibited the proteolytic activities with an IC50 of 0.39 mM for azocasein hydrolysis and of 1.35 mM for BApNA hydrolysis. Benzamidine inhibited the hydrolysis with an IC50 of 0.69 and 0.076 mM for azocasein and BApNA, respectively. The absence of cysteine-proteinases is indicated by the fact that 2-mercaptoethanol and L-cysteine did not increase the rate of azocasein hydrolysis. These results demonstrate the presence of serine-proteinases and the predominance of trypsin-like activity in the midgut of Lepidoptera insects, now also detected in A. gemmatalis, and suggest this enzyme as a major target for pest control based on disruption of protein metabolism using proteinase inhibitors.

Animals↗

Sensitivity of delta-ALA-D (E.C. 4.2.1.24) of rats to metals in vitro depends on the stage of postnatal growth and tissue.

Heavy metals, like cadmium, lead, and mercury, are potential toxic substances. The exposure to these metals can cause renal disturbances and neurological alterations. Young rats are more sensitive to harmful agents than adult animals. Delta-ALA-D enzyme acts as a biomarker of these exposures, since it has high affinity for divalent metals. The purpose of this search was to investigate the sensitivity of delta-ALA-D from suckling rats to cadmium, lead or mercury in vitro. IC(50) for delta-ALA-D activity of brain, kidneys, and liver from rats with ages between 1 and 6, 8 and 13 or 17 and 21 days was determined using metals concentrations that range from 0 to 200 microM for CdCl(2), 0 to 600 microM for HgCl(2) and from 0 to 50 microM for lead acetate. The results demonstrated that the cerebral delta-ALA-D activity is more sensitive to lead acetate than to cadmium and mercury. Delta-ALA-D from hepatic tissue is the most resistant to presence of mercury chloride in assay medium. Lead and cadmium are more toxic to renal enzyme than mercury. To sum up, the sensitivity of delta-ALA-D enzyme of young rats to heavy metals studied depends on the phase of development and tissue.

Age Factors↗

Effect of mercuric chloride and lead acetate treatment during the second stage of rapid post-natal brain growth on the behavioral response to chlorpromazine and on delta-ALA-D activity in weaning rats.

During the early post-natal period the brain is extremely sensitive to external agents. In the present study, we examined the effects of the treatment with lead acetate (3.5 or 7.0 mg/kg) and mercuric chloride (2.5 or 5.0 mg/kg) during the early post-natal period (day 8-12) on the behavioral response to chlorpromazine (CPZ) of 22-day-old rats. The effects of these metals on the sulfhydryl-containing enzyme delta-aminolevulinate dehydratase (delta-ALA-D) were also investigated. Mercuric chloride (2.5 mg/kg) did not affect brain enzyme activity, but caused a significant stimulation of renal delta-ALA-D of 24-day-old rats (27%), while animals treated with 5 mg/kg HgCl(2) showed a small but significant inhibition of cerebral (10%) and renal delta-ALA-D activity (15%). Lead acetate (3.5 or 7 mg/kg) treatment did not affect renal or cerebral delta-ALA-D. Mercuric chloride treatment (5 mg/kg) changed the pattern of open-field activity and the CPZ-induced catalepsy. However, since the undernutrition that accompanied the metal treatment also caused changes in CPZ-induced catalepsy, the effect of mercury on catalepsy could not be clearly established. Lead acetate treatment (7 mg/kg) changed the pattern of open-field motor activity and abolished the decrease in activity observed in control rats. The cataleptic response of animals to CPZ was also affected by lead acetate treatment (7 mg/kg). The increase in activity is compatible with the hyperactivity described in animals exposed to lead for long periods. Thus, the present study suggests that a short exposure to lead or mercury during suckling (second stage of rapid post-natal brain growth) caused permanent changes in locomotor activity that can be interpreted as hyperactivity. Additionally, the behavioral response to CPZ was affected by metal treatment indicating an alteration in the dopaminergic system.

Animals↗

Interference in the Limulus amebocyte lysate assay for endotoxin determination in peritoneal dialysis fluids and concentrates for hemodialysis.

The interference of the saline concentration of fluids for peritoneal dialysis and concentrates for hemodialysis on the Limulus amebocyte lysate (LAL) assay for endotoxins was investigated. The experiments were carried out individually with each substance that compose fluids for hemodialysis, to determine the possible inhibition or enhancement effects that they could cause on the LAL assay. The compositions were also assayed to investigate the possibility of synergistic effect. They were assayed by the gel-clot method from two different suppliers, and the samples that showed inhibition effect were also assayed by the chromogenic method. The samples were analysed at successive dilutions, with different LAL sensitivities, to satisfy the endotoxin limits of 5 EU/ml for the concentrate and 0.25 EU/ml for the fluid for dialysis peritoneal. The results showed that the major interference on the gel-clot assay occurs in presence of acetic acid and in concentrates containing acid acetic, even the pH being adjusted between 6.5 and 7.5. However, the test, after an adequate dilution, could be validating for all samples. Chromogenic test can be used for peritoneal dialysis fluids considering a limit of 0.25 EU/ml and sample dilution of eight times, but it cannot be used for concentrates for hemodialysis without further dilution. Considering the results and that the chromogenic is a more time-consuming method, endotoxins in fluids for hemodialysis can be satisfactorily assayed by the gel-clot method.

Dialysis Solutions↗

Parenteral nutrition and cyclosporine: do lipids make a difference? A prospective randomized crossover trial.

AIMS: This prospective, controlled, randomized crossover trial was conducted to assess the effects of parenteral nutrition, with or without lipids, in cyclosporine (CyA) pharmacokinetics. METHODS: 10 adult patients were randomized on the day of allogeneic bone marrow transplantation to receive isocaloric and isonitrogenous parenteral nutrition admixtures without (regimen A) or with lipids (regimen B). Admixtures were started on average by day + 7.4; 5 patients received regimen A followed by B, 5 in reverse order. Blood samples were collected at day 4 after transplantation, under oral diet, and 4 days after the initiation of each regimen as the sole nutrition support. At each time point, 8 whole blood samples were analysed for CyA to evaluate: area under the curve (AUC), trough concentration and systemic clearance. Clinical/laboratory events were recorded until 31 months of follow-up. RESULTS: There was no evidence of a period or treatment-by period interaction, thus results were combined for further analysis. There were no statistically significant differences between regimens in any CyA pharmacokinetic parameters; there were no significant differences from baseline values, except for a higher systemic clearance of CyA with regimen A (0.40+/-0.09 vs 0.29+/-0.06 L/Kg/h, p=0.03). CONCLUSIONS: The provision of 0.8 g/Kg/d of a 50:50 mixture of medium and long chain triglycerides did not affect CyA parameters, which were closer to baseline. In the short or long term there were no attributable side effects.

Adult↗

Reconciliation in captive Guyanese squirrel monkeys (Saimiri sciureus).

The tendency for agonistic interaction to increase the probability of friendly interaction between social partners has been demonstrated across a range of Old World primates. While research on such post-conflict behavior proceeds into an hypothesis-testing phase, new comparative information must accumulate to provide full phylogenetic perspective on primate social behavior. Data from New World and prosimian primates are yet extremely limited. We studied captive squirrel monkeys (Saimiri sciureus) via post-conflict (PC) and matched control (MC) observations and analyzed results using both the PC-MC and time-rule methods. Former opponents maintaining affiliative relationships soon engaged in friendly interaction following large proportions of agonistic interactions, whereas non-affiliated individuals, including virtually all male-female pairs, reconciled conflicts rarely. Close-proximity approaching and huddling contact constituted the principal modes of post-conflict amicability. Agonistic interactions of relatively high intensity were most likely to be reconciled and most likely to be reconciled via physical contact. High vulnerability of Saimiri to predation may have favored this species' strong inclination to reconcile soon after agonistic interaction. Research on free-living populations of this and other primate species is needed to illuminate similarities and differences across taxa.

Aggression↗

Mating season aggression and fecal testosterone levels in male ring-tailed lemurs (Lemur catta).

The challenge hypothesis (J. C. Wingfield, R. E. Hegner, B. G. Ball, and A. M. Duffy, 1990, Am. Nat. 136, 829-846) proposes that in birds, reptiles, and fish, "the frequency or intensity of reproductive aggression as an effect of T[estosterone] is strongest in situations of social instability, such as during the formation of dominance relationships, the establishment of territorial boundaries, or challenges by a conspecific male for a territory or access to mates" (p. 833). To determine the extension of this hypothesis to mammalian species, we tested predictions of the hypothesis in a nonpaternal, seasonal breeding, prosimian primate (ring-tailed lemurs, Lemur catta). Semi-free-ranging males were studied during periods of social stability (premating period) and instability (mating period). The annual mating season consists of several days during which males fight for access to promiscuous group females as each individually becomes sexually receptive for 1 day. Male rates of aggression were compared to fecal testosterone levels within premating and mating periods. In the premating period male rate of aggression was not significantly correlated with testosterone level. By contrast, during the mating season testosterone and aggression levels were positively and significantly correlated. However, on days just preceding estrus, male rate of aggression was not significantly correlated with testosterone, but on days of estrus, when aggressive challenges peaked sharply, testosterone and aggression were highly positively correlated. These results suggest that the challenge hypothesis applies to mammals as well as to birds, reptiles, and fish. In addition, elevations in testosterone were tightly circumscribed around days of estrus, suggesting a compromise between costs and benefits of elevated testosterone levels.

Aggression↗

Self and nonself stimulatory molecules induce preferential expansion of CD5+ B cells or activated T cells of chagasic patients, respectively.

It has previously been demonstrated that Trypanosoma cruzi-derived antigens (TRP) and human parasite-specific antibodies (Id) stimulate proliferation of cells from Chagasic patients. More recently, we have shown that activated T cells and CD5+ B cells are present in elevated levels in the peripheral blood of Chagasic patients. Upon in vitro exposure to these two different types of stimulatory molecules (TRP, Id), we now show that each of these elevated populations respond differentially to TRP or Id. We found that stimulation with TRP led to preferential expansion of activated T cells, while Id preferentially stimulated CD5+ B cells and CD8+ T cells. Moreover, this expansion of CD5+ B cells by Id was even more pronounced in cultures of cells from Chagasic patients with the severe, cardiac form of the disease, as compared to indeterminate patients. CD8+ T cells comprise approximately 50% of the total T cells in cultures stimulated by Id while in TRP-stimulated cultures their frequency is proportionally lower. Since parasite antigens and antiparasite antibodies are always present in the host during the chronic phase of the disease, they may also be involved with differential activation mechanisms of these cell populations in vivo.

Adult↗

Cell-specific activation of nuclear factor-kappaB by the parasite Trypanosoma cruzi promotes resistance to intracellular infection.

The transcription factor nuclear factor-kappaB (NF-kappaB) is central to the innate and acquired immune response to microbial pathogens, coordinating cellular responses to the presence of infection. Here we demonstrate a direct role for NF-kappaB activation in controlling intracellular infection in nonimmune cells. Trypanosoma cruzi is an intracellular parasite of mammalian cells with a marked preference for infection of myocytes. The molecular basis for this tissue tropism is unknown. Trypomastigotes, the infectious stage of T. cruzi, activate nuclear translocation and DNA binding of NF-kappaB p65 subunit and NF-kappaB-dependent gene expression in epithelial cells, endothelial cells, and fibroblasts. Inactivation of epithelial cell NF-kappaB signaling by inducible expression of the inhibitory mutant IkappaBaM significantly enhances parasite invasion. T. cruzi do not activate NF-kappaB in cells derived from skeletal, smooth, or cardiac muscle, despite the ability of these cells to respond to tumor necrosis factor-alpha with NF-kappaB activation. The in vitro infection level in these muscle-derived cells is more than double that seen in the other cell types tested. Therefore, the ability of T. cruzi to activate NF-kappaB correlates inversely with susceptibility to infection, suggesting that NF-kappaB activation is a determinant of the intracellular survival and tissue tropism of T. cruzi.

Animals↗

A combination of benznidazole and ketoconazole enhances efficacy of chemotherapy of experimental Chagas' disease.

Despite the large number of patients infected with Trypanosoma cruzi, there are no commercial drugs available with high efficacy for use in the clinical treatment of Chagas' disease (American trypanosomiasis). As the prospects of the introduction of new compounds by the pharmaceutical industry are poor, alternative strategies are being designed to identify candidates among drugs already available on the market that could be used in combination to provide a synergic effect and improve the efficacy of chemotherapy. In this paper we investigated a possible synergic effect of drugs in mice inoculated with isolates of T. cruzi susceptible (CL), moderately resistant (Y) and naturally resistant (Colombiana) to benznidazole and nifurtimox. Our data demonstrated that the combination of benznidazole with ketoconazole induced a synergic effect in mice infected with the CL and Y isolate of T. cruzi. No differences were observed, however, in animals infected with the Colombiana isolate, suggesting that the synergic effect of benznidazole and ketoconazole is influenced by the isolate of parasite and that this could be important in further studies searching for useful combinations of drugs. Moreover, we observed that early treatment with ketoconazole could increase the cure rate in animals infected with the Y isolate. No positive effect, in relation to cure rate, was observed with the combination of benznidazole and ofloxacin. Our results re-emphasize the importance of identifying those compounds already on the market with synergic effects able to enhance the cure of T. cruzi infection.

Animals↗

Mediation of Cryptosporidium parvum infection in vitro by mucin-like glycoproteins defined by a neutralizing monoclonal antibody.

The protozoan parasite Cryptosporidium parvum is a significant cause of diarrheal disease worldwide. Attachment to and invasion of host intestinal epithelial cells by C. parvum sporozoites are crucial steps in the pathogenesis of cryptosporidiosis. The molecular basis of these initial interactions is unknown. In order to identify putative C. parvum adhesion- and invasion-specific proteins, we raised monoclonal antibodies (MAbs) to sporozoites and evaluated them for inhibition of attachment and invasion in vitro. Using this approach, we identified two glycoproteins recognized by 4E9, a MAb which neutralized C. parvum infection and inhibited sporozoite attachment to intestinal epithelial cells in vitro. 4E9 recognized a 40-kDa glycoprotein named gp40 and a second, >220-kDa protein which was identified as GP900, a previously described mucin-like glycoprotein. Glycoproteins recognized by 4E9 are localized to the surface and apical region of invasive stages and are shed in trails from the parasite during gliding motility. The epitope recognized by 4E9 contains alpha-N-acetylgalactosamine residues, which are present in a mucin-type O-glycosidic linkage. Lectins specific for these glycans bind to the surface and apical region of sporozoites and block attachment to host cells. The surface and apical localization of these glycoproteins and the neutralizing effect of the MAb and alpha-N-acetylgalactosamine-specific lectins strongly implicate these proteins and their glycotopes as playing a role in C. parvum-host cell interactions.

Animals↗

Serum levels of cytokines in patients envenomed by Tityus serrulatus scorpion sting.

Seventeen patients stung by Tityus serrulatus scorpion were classified as mild (pain at the site of the sting, n = 6), moderate (local pain and one of the following manifestations: vomiting, psychomotor agitation, prostration, sweating, tachypnea, tachycardia and mild arterial hypertension, n = 10) and severe cases (equal moderate cases plus cardiac failure, pulmonary edema and shock, n = 1). Venous blood was sampled for biochemical and hematological analysis and for IL-1alpha, IL-6, IL-10, TNF-alpha, IFN-gamma and GM-CSF ELISAs at the time of hospital admission, 6 h (moderate and severe cases), and 12, 18, 36 and 72 h (severe case) later. Ten age-matched healthy volunteers were used as control. Increased serum levels of IL-1alpha was noticed in all patients, high levels of IL-6, IFN-gamma and GM-CSF were observed only in a patient with severe envenomation. Our data suggest that a systemic inflammatory response-like syndrome is triggered during severe envenomation caused by T. serrulatus sting and that release of cytokines may be involved in this response.

Blood Cell Count↗