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Biomedical subjects

C A Pereira

Publications and source records attributed to C A Pereira.

At least 19 recordsLinked to original sources

Biomechanical and ultrastructural comparison of cryopreservation and a novel cellular extraction of porcine aortic valve leaflets.

Heart valve substitutes of biological origin often fail by degenerative mechanisms. Many authors have hypothesized that mechanical fatigue and structural degradation are instrumental to in vivo failure. Since the properties of the structural matrix at implantation may predetermine failure, we have examined the ultrastructure, fracture, mechanics, and uniaxial high-strain-rate viscoelastic properties of: (1) fresh, (2) cryopreserved, and (3) cellular extracted porcine aortic valve leaflets. The cellular extraction process is being developed in order to reduce immunological attack and calcification. Cryopreservation causes cellular disruption and necrotic changes throughout the tissue, whereas extraction removes all cells and lipid membranes. Both processes leave an intact collagen and elastin structural matrix and preserve the high-strain-rate viscoelastic characteristics of the fresh leaflets. Extraction does cause a 20% reduction in the fracture tension and increases tissue extensibility, with the percent strain at fracture rising to 45.3 +/- 4 (mean +/- SEM) from 31.5 +/- 3 for fresh leaflets. However, extraction does preserve matrix structure and mechanics over the physiological loading range. Glutaraldehyde fixation produces increased extensibility, increased elastic behavior, and, when applied to extracted leaflets, it causes a marked drop in fracture tension, to 50% of that for fresh leaflets. The combination of extraction and fixation may lead to early degenerative failure. The cellular extraction technique alone may be a useful alternative to glutaraldehyde fixation in preparing bioprosthetic heart valves.

Animals

Interferon-gamma levels during the course of Trypanosoma cruzi infection of Calomys callosus (Rodentia-Cricetidae) and Swiss mice.

Serum levels of interferon-gamma (IFN-gamma) were evaluated in Calomys callosus and Swiss mice during the course of infection by four strains of Trypanosoma cruzi. All strains stimulated the production of this interleukine; however, the timing of its onset and permanence varied among strains and between the two animal models. When chronically infected animals with no detectable serum IFN-gamma were challenged with the homologous strain, they produced quantities comparable with those obtained during the acute phase of infection. In C. callosus there was a correlation between H2O2 liberation by peritoneal macrophages and serum IFN-gamma levels, whereas no such correlation was found in mice. C. callosus had a higher capacity to heal histopathological lesions, whereas lesions in mice were progressive. The results obtained suggest that C. callosus develops well-adapted immune mechanisms that may be important for its role as a reservoir of T. cruzi.

Animals

Specific T-cell response correlates with resistance of genetic heterogeneous mouse populations to mouse hepatitis virus 3 infection.

In a recently published study [Vassão RC, Mello IGC, Pereira CA (1994) Arch Virol 137: 277-288] we have shown that the genetically selected high antibody responder mice (HIII) are susceptible and the low antibody responder counterparts (LIII) are resistant to death induced by experimental infection with mouse hepatitis virus 3 (MHV3). This report shows that the MHV3 titers in the peritoneal exudate (PE) of HIII mice, 3 days after infection, were more than 2 log greater than in the resistant LIII mice, the interferon gamma (IFN gamma) titers in the PE of both mouse populations being not significantly different. The treatment with monoclonal antibodies (mAb) against CD4+ or CD8+ T cells induced susceptibility among LIII mice. The depletion of CD4+ T-cell subset in LIII mice was evidenced by, and led to a significant reduction in, the IFN gamma synthesis in their PEs with a 100 fold increase in MHV3 titers. When lymph node cells (LNC) were harvested from MHV3-infected mice and stimulated "in vitro" with MHV3 inactivated by ultraviolet radiation (uv-MHV3), only LNC from LIII mice were capable of proliferating and synthesizing significant amounts of interleukin 2 (IL-2). The LNC proliferation and IL-2 synthesis were inhibited by treatment with mAbs against CD4 or CD8 molecules. The MHV3 infection induced in both lines of mice a profound depression of the mitogenic response of LNC to phytohemaglutinin (PHA). A correlation between the specific T-cell response and the resistance to MHV3 infection is discussed.

Animals

A multi-sample denaturation temperature tester for collagenous biomaterials.

The temperature at which collagen denatures from a triple helix to a random coil structure is a useful measure of the degree of crosslinking. A new multi-sample denaturation temperature tester (DTT) has been constructed for rapid determination of the collagen denaturation temperature of natural tissues and collagenous biomaterials. To validate the system, the denaturation temperatures measured for the DTT are compared with results from differential scanning calorimetry (DSC). Data are presented for bovine pericardium in three states with denaturation temperatures ranging from 68 to 85 degrees C: fresh, or crosslinked with glutaraldehyde or the epoxide reagent Denacol EX-512 poly (glycidyl ether). Denaturation temperatures measured by DTT were not significantly different from those measured by differential scanning calorimetry (DSC); however, DSC onset systematically occurred at a slightly lower temperature than that measured by DTT. This result, seen only for fresh tissue is in agreement with earlier experiments using hydrothermal isometric tension (HIT) testing. By contrast, DTT and DSC onset were identical for the exogenously crosslinked materials. Since the measured transition temperature was independent of initial load, this variable may be chosen to yield sharper force-temperature transitions with a given sample geometry. This instrument allows accurate assessment of collagen denaturation temperatures for multiple samples in a fraction of the time required by other methods.

Animals

Outcome and quality of life of patients with severe chronic limb ischaemia: a cohort study on the influence of diabetes.

OBJECTIVE: To determine the influence of diabetes on the use of arterial reconstruction, the rate of amputation and death, and the quality of life of patients with severe limb ischaemia. DESIGN: A prospective study of patients with the first episode of ischaemia. SETTING: University tertiary referral centre. METHODS: Thirty-seven patients with diabetes and 50 without diabetes, were studied over a 12 month period with complete follow-up. MAIN OUTCOME MEASURES: The proportion of patients undergoing an arterial reconstruction, amputation rate, death rate, and quality-of-life scores. RESULTS: Patients with diabetes underwent an arterial reconstruction less often than patients without diabetes (7/37 vs. 18/50). The odds of patients with diabetes having a higher incidence of adverse outcome was 1666:1 for minor amputation, 26:1 for major amputation, and 4.7:1 for death. There was a tendency towards a lower quality of life for patients with diabetes at 3 (OR 1.94, p = 0.036), 6 (OR 1.58, p = 0.117), and 12 (OR 1.47, p = 0.185) months. CONCLUSIONS: In patients with diabetes, (1) the opportunity of undergoing an arterial reconstruction is lower, (2) morbidity and mortality are higher, and (3) the quality of life tends to be worse.

Activities of Daily Living

Incidence of and risk factors for hepatitis B virus and hepatitis C virus infection among haemodialysis and CAPD patients: evidence for environmental transmission.

Hepatitis B virus (HBV) serum markers (HBsAg, anti-HBs, anti-HBc) and antihepatitis C antibody (anti-HCV) were prospectively followed in haemodialysis and CAPD patients. From January 1987 to January 1990, 185 patients on haemodialysis and 124 on CAPD were analysed. Among patients susceptible to HBV (69 on haemodialysis and 70 on CAPD), there were 17 HBsAg seroconversions on haemodialysis (0.19/patient-year) and 1 on CAPD (0.01/patient-year). A Cox proportional hazards model showed that haemodialysis treatment was the only risk factor significantly associated with HBV infection, thus suggesting transmission through the environment. Regarding hepatitis C, 83 anti-HCV-negative patients on haemodialysis and 46 on CAPD were followed. There were 18 seroconversions on haemodialysis (0.15/patient-year) and two seroconversions on CAPD (0.03/patient-year). Haemodialysis treatment was also the only risk factor significantly associated with a higher risk of HCV infection. The hazard ratio for HCV infection in haemodialysis patients was 5.7 compared to CAPD patients. Nevertheless, for one patient on CAPD treatment transfusions were the only possible source of HCV infection. In conclusion, both viruses were transmitted mainly through the haemodialysis environment, but the role of transfusions could not be excluded.

Adolescent

Mouse hepatitis virus type 3 infection provokes a decrease in the number of sinusoidal endothelial cell fenestrae both in vivo and in vitro.

Fenestrations of hepatic endothelial cells play an active role as a sieving barrier allowing extensive exchange between the blood and liver parenchyma. Alteration of these structures may be induced in the course of various pathological events and provoke important perturbations of liver function. We demonstrate here that sinusoidal endothelial cells are permissive for mouse hepatitis virus 3 (MHV3) in vivo and in vitro and that this infection leads to a striking decrease in the number of fenestrae. The disappearance of these structures observed under scanning electron microscopy or in cryofracture preparations in vivo and in vitro cannot be reversed by the action of cytochalasin B on the microfilament network. The decrease in the porosity seems to be related directly to the productive infection of the endothelial cells, because it was not observed in A/J mice resistant to the virus and in susceptible BALB/c mice immunized with a thermosensitive mutant in which no viral replication occurs. In conclusion, a viral infection of liver endothelial cells may cause extensive loss of the fenestrations and thus lead to important functional pertubations.

Animals

Neutralization of the effect of Crotalus durissus terrificus venom by gangliosides.

We determined the ability of a mixture of gangliosides (16% GD1b, 19% GT1b, 21% GM1, 40% GD1a) to neutralize the effect of Crotalus durissus terrificus (Cdt) venom in vitro and in vivo. Protection was indicated by the absence of muscular contractions, hind limb paralysis or death of BALB/c mice (16-18 g) after receiving Cdt venom (1 microgram Cdt venom containing 0.6 microgram protein) at the doses indicated. A dose of Cdt venom above 0.9 microgram (ip) or 1 microgram (im) induced muscular contraction and above 1.2 micrograms (ip) or 5.5 micrograms (im) the venom induced muscular contraction and hind limb paralysis. Cdt venom above 2.5 micrograms (ip) or 9 micrograms (im) induced all these symptoms and 95 to 100% death in experimental animals. The lethal dose 50% of the Cdt venom used was 8 micrograms (im) and 1.5 micrograms (ip). In in vitro studies, 4 mg gangliosides neutralized the effect of up to 1.5 micrograms Cdt venom. Quantities as low as 0.2 mg gangliosides were capable of neutralizing 0.9 microgram of Cdt venom in vitro. Intramuscular treatment with 1 mg gangliosides performed 60 min after the intramuscular injection of 5 micrograms Cdt venom protected 100% of the animals. In contrast, no protection was achieved with intraperitoneal treatment with gangliosides. The data show that gangliosides were effective in neutralizing the toxic effects induced by Crotalus durissus terrificus venom both in vitro and in vivo and that post-exposure intramuscular treatment with gangliosides could protect animals experimentally inoculated with the venom.

Animals

Development of a pericardial acellular matrix biomaterial: biochemical and mechanical effects of cell extraction.

There is evidence to suggest that the cellular components of homografts and bioprosthetic xenografts may contribute to calcification or immunogenic reactions. A four-step detergent and enzymatic extraction process has been developed to remove cellular components from bovine pericardial tissue. The process results in an acellular matrix material consisting primarily of elastin, insoluble collagen, and tightly bound glycosaminoglycans. Light and electron microscopy confirmed that nearly all cellular constituents are removed without ultrastructural evidence of damage to fibrous components. Collagen denaturation temperatures remained unaltered. Biochemical analysis confirmed the retention of collagen and elastin and some differential extraction of glycosaminoglycans. Low strain rate fracture testing and high strain rate viscoelastic characterization showed that, with the exception of slightly increased stress relaxation, the mechanical properties of the fresh tissue were preserved in the pericardial acellular matrix. Crosslinking of the material in glutaraldehyde or poly(glycidyl ether) produced mechanical changes consistent with the same treatments of fresh tissue. The pericardial acellular matrix is a promising approach to the production of biomaterials for heart valve or cardiovascular patching applications.

Animals

Effect of molecular structure of poly(glycidyl ether) reagents on crosslinking and mechanical properties of bovine pericardial xenograft materials.

With the identification of the exacerbating effect of glutaraldehyde on calcification of heart valve materials, there exists a renewed interest in both alternative reagents and the effects of crosslinking on connective tissues. One potentially useful class of reagents are poly(glycidyl ether) compounds. We have examined 5 of these reagents with different molecular sizes and functionalities for their effects on mechanical properties and collagen denaturation (shrinkage) temperature. Samples of bovine pericardium were tested fresh or after 48 h fixation in one of the five compounds for denaturation temperature, stress-strain response, stress relaxation, plastic deformation, and fracture properties. Of the compounds tested, those with intermediate length backbones and 4 or 5 epoxide groups were most effective in producing intrahelical crosslinking and increased denaturation temperature over 48 h. However, in samples examined after 17 months of fixation, all reagents had equivalently increased the denaturation temperature. Examination of mechanical results revealed two distinct mechanisms for mechanical change. Observed shifting of the stress-strain curve to the right (due to shrinkage), increased plastic deformation, and some reduction of stress relaxation are all unrelated to denaturation temperature (and hence to changes in intrahelical crosslinking). An alternate mechanism, perhaps formation of intermolecular crosslinks may be responsible. Intrahelical crosslinking produces only lesser reductions in stress relaxation. Cross-comparison of reagents of differing molecular structure provides a useful tool toward increased understanding of the mechanical consequences of tissue crosslinking.

Animals

Role of macrophages, interferon gamma and procoagulant activity in the resistance of genetic heterogeneous mouse populations to mouse hepatitis virus infection.

Genetic heterogeneous mouse populations selected for high (HIII) and low (LIII) antibody response were used to study some aspects of mouse hepatitis virus 3 (MHV3) infection, such as the resistance pattern, virus replication in the liver and peritoneal exudate or in cultured peritoneal macrophages, the interferon (IFN) synthesis in the serum and peritoneal exudate and the procoagulant activity (PCA) of the peritoneal exudate (PEC) and spleen cells (SC). The HIII mice, when compared to their LIII mice counterparts, were susceptible to MHV3 infection showing higher virus titres in the liver and peritoneal exudate, comparable IFN alpha/beta or IFN gamma titres in the peritoneal exudate or in the serum, and higher levels of PCA of PEC and SC. A higher virus titre was detected in the supernatants of HIII mouse macrophages infected with MHV3. The activation of HIII mouse macrophages with LPS, IFN alpha/beta or IFN gamma, in contrast to that of LIII mouse macrophages, did not induce an antiviral effect with partial restriction of the MHV3 replication. The LPS antiviral activity was shown to be partially exerted by IFN alpha/beta synthesis. The IFN gamma was shown to be more effective in inducing an antiviral state in LIII macrophages, when compared to IFN alpha/beta. The data obtained are consistent with the notion that the resistance mechanisms to the MHV3 infection involve the PCA and the sensitivity of macrophages to IFN.

Animals

A genetic analysis of macrophage activation and specific antibodies in relation to the resistance of heterogeneous mouse populations to MHV3 infection.

The genetically selected high antibody responder mice (HIII) are susceptible and the low antibody responder mice (LIII) are resistant to the experimental infection with Mouse Hepatitis Virus 3 (MHV3). The mortality rates of the F1 hybrids and of the F2 segregants showed the codominance of the susceptible and resistant characters. The direct individual intrapopulation correlation between the induction of antiviral state in macrophages activated by IFN gamma and the resistance to the virus infection, showed that an antiviral state could be induced in resistant mouse macrophages, whereas in susceptible mouse macrophages no restriction of virus replication could be observed. A direct inter- and intrapopulation correlation of pre-existing antibody titres against MHV3 with the mortality and a direct interpopulation correlation of those titres with the mean survival time of susceptible animals was shown. The data indicate, among the mechanisms of resistance against the virus infection, a role of IFN gamma macrophage-activation and of antibodies against MHV3 which may delay the mean survival time in susceptible animals.

Animals

Biomonitoring of nurses handling antineoplastic drugs.

The micronuclei analysis in exfoliated cells of the buccal cavity was employed in the cytogenetic monitoring of nurses handling antineoplastic drugs. The group under study consisted of 25 subjects who showed a marked increase in micronucleated cells as compared with the control group (Chi-square = 15.12, with one degree of freedom, P < 0.001).

Adult

Gamma-IFN and macrophage respiratory burst in Calomys callosus challenged with Trypanosoma cruzi bloodstream and metacyclic forms.

Parasitemia levels of Calomys callosus inoculated with a high dose (HBT) of 4 x 10(3) Trypanosoma cruzi strain M226 bloodstream trypomastigotes (BT) exceeded those with the same inoculum of metacyclic trypomastigotes (MT) while a similar parasitemia was obtained with a low dose (LBT) of 5 x 10(2) of BT. Serum IFN-gamma levels during the acute phase of infection were higher in the LBT inoculated group when compared with the group inoculated with HBT, while the IFN-gamma levels in MT inoculated animals were close to uninfected controls. Spontaneous liberation of H2O2 of peritoneal macrophages explanted from animals on days 21 and 28 after infection was comparable to that of controls for HBT and LBT groups while that of the MT inoculated group was significantly higher. Phorbol Myristate Acetate (PMA) stimulation resulted in high H2O2 liberation specially in the infected groups. In vitro challenge with BT suppressed the small amount of spontaneous H2O2 release, while MT challenge stimulated this release to a limited degree in infected groups. In this animal model, interacting with a parasite strain isolated from the same host, macrophage activation as measured by H2O2 release was low, while the same strain had been previously observed to result in hyperactivation of mouse macrophages. We suggest that this distinctive behavior may be due to a host-parasite adaptation.

Animals

Psychiatric manifestations of systemic lupus erythematosus: clinical features, symptoms, and signs of central nervous system activity in 43 patients.

Forty-three female inpatients with active systemic lupus erythematosus (SLE) were studied by a multidisciplinary team to answer the following research questions: 1) What are the features of the psychopathology in patients with active SLE? and 2) In these patients, what is the relationship between psychiatric disorders and symptoms and signs suggesting activity of SLE in the CNS? Our a priori hypothesis was that, in patients with active SLE, those with psychiatric manifestations would have more symptoms and signs of CNS activity than those without psychiatric manifestations. Psychiatric evaluation consisted of standardized psychiatric instruments and diagnostic criteria. The assessment of SLE systemic and central nervous system (CNS) activity consisted of rheumatologic, neurologic, and ophthalmologic evaluations; serum and cerebral spinal fluid (CSF) analysis; brain computerized tomography (CT); and electroencephalogram (EEG). Twenty-seven patients (63%) presented psychiatric symptoms (Psychiatric Group), and 16 (37%) patients presented no current psychiatric diagnosis (Nonpsychiatric Group). These groups were compared in terms of the above variables. Depressive syndrome was the most frequent diagnosis (44%) followed by delirium (7%) and dementia (5%). Psychiatric symptoms were associated with subjective cognitive impairment (85%) and neurologic abnormality (85%). Widened cortical sulci was the most frequent CT alteration and was equally common in both groups. No statistical difference was found between the 2 groups regarding their general clinical evaluation, serum and CSF exams, or EEG alterations. To determine whether the severity of psychiatric symptoms was related to CNS activity, we divided the 27 patients with psychiatric manifestations into 2 groups: the Major Group--18 patients with major psychopathology, and the Minor Group--9 patients with mild depressive syndromes.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Poisoning by the use of Datura leaves in a homemade toothpaste.

Datura stramonium and related species are relatively common causes of atropine-like poisoning by ingestion or inhalation. Toxic absorption after mucosal application is evident in 24 h of atropinism sustained by a woman who used a toothpaste mixed with the leaves and flowers of Datura sp., table salt, vinegar and an alcoholic beverage.

Absorption

Relationship of interferon synthesis and the resistance of mice infected with street rabies virus.

Genetically homogeneous and heterogeneous mouse populations were tested for resistance to experimental street rabies virus infection and their ability to synthesize interferon (IFN) during the infection. The genetically heterogenous HI mouse population was highly resistant (12% mortality), and the genetically homogeneous BALB/c and C3H mice as well as the genetically heterogeneous Sw and LI mouse populations were susceptible (60 to 71% mortality). The genetically homogeneous A/J mice were highly susceptible (85% mortality) to experimental street rabies infection. The ability of these mice to synthesize IFN as measured in serum 4 days after the infection was directly related to the degree of resistance, with the highly resistant HI mice showing large amounts of IFN (850 U/ml), and the susceptible mice showing low amounts of IFN (50 to 280 U/ml). IFN induced within the central nervous system and measured in brain homogenates during infection was not correlated with resistance. The present data suggest that high levels of IFN occurring in serum early during infection with street rabies virus contribute to the resistance of these mice.

Animals