Ana Maria Aparecida Guaraldo

Transmission of Chagas Disease (American Trypanosomiasis) by Food

In April 2009, the centenary of the discovery of the American trypanosomiasis, or Chagas disease, was celebrated. A hundred years after the discovery, little has been invested in diagnostics and treatment because the disease affects mainly poor people in developing countries. However, some changes in the epidemiology of the disease are of great importance today. Chagas disease transmitted through food is a public health concern in all areas where there is a reservoir of Trypanosoma cruzi in wild animals (e.g., mammals and marsupials) and/or where infected triatomine bugs are in contact with human food source items (especially fruits and vegetables). Recently, several outbreaks of illness related to the ingestion of food contaminated with T. cruzi have been recorded in Brazil, Colombia, and Venezuela.

Evolução dos esporocistos de Schistosoma mansoni Sambon, 1907 em Biomphalaria glabrata (Say, 1818) e Biomphalaria tenagophila (D'Orbigny, 1835)

Estudou-se a evolucao dos esporocistos de Shistosoma mansoni das linhagens BH e SJ respectivamente em Biomphalaria glabrata e Biomphalaria tenagophila. Utilizando-se cortes histologicos foram avaliados o aspecto e numero de esporocistos primarios desde a primeira ate a oitava semana de infeccao, a contar do dia em que cada molusco foi exposto a 100 miracidios. No decorrer da primeira semana constataram-se diferencas significativas entre as linhagens estudadas quanto ao numero e aspecto dos esporocistos primarios, A distribuicao por orgaos e a evolucao dos esporocistos foi observada ate a fase de formacao das cercarias infectantes.

Chromatin supraorganization and extensibility in mouse hepatocytes with development and aging.

Chromatin supraorganization and extensibility, which lead to the formation of extended chromatin fibers (ECF), are affected by starvation and refeeding in adult mouse hepatocytes. It is expected that they could also change with mouse development and aging.

Chromatin supraorganization and extensibility in mouse hepatocytes following Starvation and refeeding

The effect of 48 h of starvation and of 48 h of refeeding subsequent to starvation on chromatin supraorganization and extensibility was studied in hepatocytes from adult mice.

Parasite survey in mouse and rat colonies of Brazilian laboratory animal houses kept under differents sanitary barrier conditions

A parasitological study was undertaken to determine the health status of 15 mouse and 10 rat colonies bred in 18 Brazilian laboratory animal houses maintained under different sanitary barrier conditions which supply animals for teaching, research purposes and manufacture of biological products for medical or veterinary use. Parasitological methods were used for diagnosis of mites, lices, helminthes and protozoan parasites. A questionnaire was answered by institutions with the intention to obtain information about the existence of barriers against infections and of regular sanitary monitoring program of their colonies. The questionnaire data show that the majority of the animal houses investigated do not possess an efficient sanitary barrier system able to keep animals under controlled health sanitary conditions. Ecto and endoparasite infections are widespread in the colonies and multiple infections were common in animals from most facilities investigated. The prevalences of parasites detected among the mouse and rat colonies of the laboratory animal houses investigated were: Myocoptes musculinus (46.6%), Myobia musculi (26.6%), Radfordia ensifera (13.3%), Syphacia obvelata (86.6%), Aspiculuris tetraptera (60.0%), Hymenolepis nana (53.3%), Spironucleus muris (80.0%), Tritrichomonas muris (80.0%), Giardia muris (66.0%), Entamoeba muris (20.0%), Eimeria sp. (13.3%), Hexamastix muris (26.6%), Poliplax spinulosa (30.0%), Poliplax serrata (10.0%), Radfordia ensifera (30.0%), Syphacia muris (80.0%), Hymenolepis nana (40.0%), Trichosomoides crassicauda (55.5%), Spironucleus muris (90.0%), Tritrichomonas muris (80.0%), Giardia muris (60.0%), Entamoeba muris (80.0%), Eimeria sp. (60.0%) and Hexamastix muris (60.0%).

Enhancement of natural killer cell function by titanocenes in mice bearing Ehrlich ascites tumour

In the present work, we studied the effects of two titanocenes, biscyclopentadienyldichlorotitanium IV, (DDCT) and its derivative, biscyclopentadienylditiocianatetitanium IV (BCDT), on the activity of natural killer (NK) cells in Ehrlich ascites tumour (EAT)-bearing BALB/c mice. In order to investigate a more direct effect of these compounds on NK cell function, we performed experiments with severe combined immunodeficiency (SCID) mice, which exhibit a normal NK cell response in the absence of T and B cells. The treatment consisted of intraperitoneal (i.p.) administration of 15 mg/kg/day of DDCT for 2 days or 10 mg/kg/day of BCDT for 3 days. In addition, to verify whether the effects produced by the titanocenes were compound specific or related to a direct antitumour effect, we also investigated the effects of a 3-day treatment with 100 mg/kg of cyclophosphamide cyclophosphamide on NK cell activity. Our results demonstrated that, in BALB/c and SCID mice, NK cell function declined to subnormal levels after inoculation of the tumour. In these animals, although treatment with DDCT and BCDT significantly enhanced NK cell function, only DDCT restored NK cell activity to normal values in all stages studied. Conversely, treatment with cyclophosphamide reduced NK cell function in nontumour bearing SCID mice and was also unable to restore the decreased NK activity of tumour-bearing SCID mice, thus demonstrating that the enhancement of NK cell function by titanocenes is compound specific. The same effect of cyclophosphamide was observed with BALB/c mice. In the present study, the up-modulatory effects of these two compounds on NK cell function reveal a new aspect of the mechanism of antitumoural action of titanocenes.

DNA content, chromatin supraorganization, nuclear glycoproteins and RNA amounts in hepatocytes of mice expressing insulin-dependent diabetes.

Chromatin supraorganization and extensibility and nuclear glycoprotein content have been reported to change in hepatocytes from mice during development and aging, as well as under starvation and refeeding conditions. In non-obese diabetic (NOD) mice, the expression of insulin-dependent diabetes may be accompanied by metabolic changes in the liver. These changes are likely to be similar to those involved in the aging processes of non-diabetic animals. Therefore, we hypothesized that the chromatin organization, as well as the physical properties and compositions of hepatocyte nuclei would also be affected in NOD mice in the same way as those in aged non-diabetic mice. Nuclear image parameters were evaluated by image analysis of Feulgen-stained preparations. Chromatin extensibility in response to gravity was observed with polarized light after lysis and toluidine blue staining. The Con-A response of nuclear glycoproteins was evaluated with scanning microspectrophotometry. These characteristics were assessed using hepatocyte imprints from female NOD mice after a 28-day period of diabetes expression. Observations and measurements were made in comparison to healthy BALB/c mice. Total RNA amounts were determined for livers of NOD and BALB/c mice. Enhanced polyploidy levels, a decrease in chromatin higher-order packing states, an increased frequency of extended chromatin fiber formation, and deeper Con-A-responsive chromatin areas were observed in the hepatocytes of the NOD mice expressing insulin-dependent diabetes. Reduced amounts of total RNA were also found in the livers of these mice. Our findings for NOD mice expressing insulin-dependent diabetes are consistent with previously reported data for old-aged mice of the inbred strain A/Uni and may reflect changes in transcriptional activities associated with the stressful physiological demands on the liver during the expression of diabetes.

Survival In Vitro and Virulence of Trypanosoma cruzi in Acai Pulp in Experimental Acute Chagas Disease

Chagas disease is a parasitic infection with high socioeconomic impact throughout Latin America. Although this severe, incurable disease can be transmitted by several routes, oral transmission is currently the most important route in the Amazon Basin. Açaí pulp has nutritional properties and is popular throughout Brazil and abroad. However, this pulp has been associated with microepidemics of acute Chagas disease (ACD) in northern Brazil, where açaí fruit is the main food supplement. In this study, we examined the in vitro survival and in vivo virulence of Trypanosoma cruzi Y strain in açaí pulp. Aliquots of in natura açaí pulp produced in Belém city in the northern Brazilian state of Pará were mixed with 10⁵ trypomastigotes. The samples were incubated at room temperature or at 4 or -20°C for various periods, and the parasites were isolated by forced sieving. The resulting eluates were examined by microscopy, and the trypomastigotes were administered intraperitoneally, orally, or by gavage to immunodeficient mice (C.B-17-Prkdc(scid)/PasUnib) that had been pretreated with antibiotics. Parasitemia was quantified by the Brener method, and mortality was recorded daily. All routes of administration resulted in ACD. A 5-day delay in the onset of parasitemia occurred with oral administration. The survival and virulence of the parasites were unaffected by prior incubation at room temperature for 24 h, at 4°C for 144 h, and at -20°C for 26 h. These results indicate that T. cruzi can survive and retain its virulence in açaí pulp under various conditions and that cooling and freezing are not suitable methods for preventing foodborne ACD.

Esquistossomose mansônica em camundongos experimentalmente subnutridos

Mansoni schistosomiasis was studied in mice fed on a low protein diet. Four groups of the Swiss breed Mus musculus were used in an experiment with two factors, each with two levels: 1-non-infected, normal diet; 2 - infected, normal diet ; 3 - non-infected, low protein diet; 4 - infected, low protein diet. The mice were killed for observation at age 90 days, after 60 days of infection, for those infected. It was found that the worms suffered the effects of malnutrition, mainly males, whose population count was cut by half, in addition to poor individual growth. The hepatic granuloma count was found to be smaller in the undernourished group; while the corresponding lesions were also generally smaller. There was a marked leukopenia in the malnourished mice, even more so in the infected mice. The severe lymphopenia and eosinopenia observed suggest that the immunologic system of the host was affected by malnutrition. Mortality rates were far larger in the infected, low protein diet group. One may then surmise that the malnourished mice offered lower resistance to the infection, their lower number of granuloma notwithstanding.

Nuclear phenotypes and DNA fragmentation in tendon fibroblasts of NOD mice

Abstract Changes in DNA/chromatin structure, ploidy degrees and cell death possibly caused by oxidative stress during the insulin-dependent diabetes have been reported for different cell types. However, all these studies have been carried in streptozotocin-induced diabetic rats or mice and showed contradictory results. In this work, nuclear phenotypes and DNA fragmentation were investigated in fibroblasts from mice spontaneously developing insulin- dependent diabetes (NOD) and compared with healthy (BALB/C) mice. Geometric, densitometric and textural parameters obtained for Feulgen-stained nuclei by image analysis were used to define nuclear phenotypes. Significant differences were observed for nuclear sizes and for densitometric and textural parameters of the tendon nuclei. Optical density, Feulgen-DNA values, transmittance variability per nucleus and nuclear entropy values were significantly higher in the NOD mice. The Feulgen-DNA amounts for the NOD and BALB/C mice were found to be distributed into several doubling Feulgen-DNA classes. The frequency of nuclei with the smallest Feulgen-DNA amounts, which may represent DNA fragmentation and loss, was lower in fibroblasts of the NOD mice (2.3%) in comparison to the BALB/C mice (38%). In contrast, the frequency of polyploid nuclei in NOD mice was higher (24.5%) than that in BALB/C mice (1.9%). Based on optical density, transmittance variability per nucleus, and nuclear entropy data, a larger contrast between highly and less densely packed states, was demonstrated for the fibroblasts of the NOD mice. Maybe the deeper condensation of the highly packed chromatin evident in NOD fibroblasts is related to silencing of some genes involved with changes in tendon supraorganization with the diabetes.