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The fate of three pesticides (vinclozolin, dimethoate, cyproconazole) in plums, from field treatment to the drying process, was studied. Only vinclozolin showed measurable residue concentrations at harvest, while dimethoate was completely degraded after two weeks and cyproconazole was present at negligible levels just after treatment. During the drying process into prunes, the residues were not reduced during the fruit washing stage, but the drying stage led to complete elimination of vinclozolin residues.

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As demonstrated previously in our laboratory, B-1 cells migrate from the peritoneal cavity of mice and home to a distant site of inflammation to become macrophage-like cells. However, the influence that these cells might have on the kinetics and fate of the inflammatory process is not known. Considering that macrophages are pivotal in the inflammatory reaction, we decided to investigate the possible influence B-1 cells could have on macrophage activities in vitro. Our results show that peritoneal macrophages from Xid mice, a mouse strain deprived of B-1 cells, have higher phagocytic indexes for zymozan particles when compared with macrophages from wild-type mice. Moreover, macrophages from wild-type mice have a lower ability to release nitric oxide and hydrogen peroxide when compared with macrophages from Xid mice. Experiments using cocultures of B-1 cells and macrophages from Xid mice in transwell plates demonstrated that B-1 cells down-regulate macrophage activities. These observations also indicate that this phenomenon is not due to a physical interaction between these two cell populations. As B-1 cells are one of the main sources of interleukin (IL)-10, we demonstrate in this study that adherent peritoneal cells from Xid mice produce significantly less amounts of this cytokine in culture when compared with IL-10 production by cells from wild-type mice. When B-1 cells from IL-10 knock-out mice and macrophages from wild-type mice were cocultured in transwell plates, the phagocytic index of macrophages was not altered demonstrating that B-1 cells can influence the effector functions of macrophages in vitro via IL-10 secretion.

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Heme is present in all cells, acting as a cofactor in essential metabolic pathways such as respiration and photosynthesis. Moreover, both heme and its degradation products, CO, iron and biliverdin, have been ascribed important signaling roles. However, limited knowledge is available on the intracellular pathways involved in the flux of heme between different cell compartments. The cattle tick Boophilus microplus ingests 100 times its own mass in blood. The digest cells of the midgut endocytose blood components and huge amounts of heme are released during hemoglobin digestion. Most of this heme is detoxified by accumulation into a specialized organelle, the hemosome.We followed the fate of hemoglobin and albumin in primary cultures of digest cells by incubation with hemoglobin and albumin labeled with rhodamine. Uptake of hemoglobin by digest cells was inhibited by unlabeled globin, suggesting the presence of receptor-mediated endocytosis. After endocytosis, hemoglobin was observed inside large digestive vesicles. Albumin was exclusively associated with a population of small acidic vesicles, and an excess of unlabeled albumin did not inhibit its uptake. The intracellular pathway of the heme moiety of hemoglobin was specifically monitored using Palladium-mesoporphyrin IX (Pd-mP) as a fluorescent heme analog. When pulse and chase experiments were performed using digest cells incubated with Pd-mP bound to globin (Pd-mP-globin), strong yellow fluorescence was found in large digestive vesicles 4 h after the pulse. By 8 h, the emission of Pd-mP was red-shifted and more evident in the cytoplasm, and at 12 h most of the fluorescence was concentrated inside the hemosomes and had turned green. After 48 h, the Pd-mP signal was exclusively found in hemosomes. In methanol, Pd-mP showed maximal emission at 550 nm, exhibiting a red-shift to 665 nm when bound to proteins in vitro.The red emission in the cytosol and at the boundary of hemosomes suggests the presence of heme-binding proteins, probably involved in transport of heme to the hemosome. The existence of an intracellular heme shuttle from the digestive vesicle to the hemosome acting as a detoxification mechanism should be regarded as a major adaptation of ticks to a blood-feeding way of life. To our knowledge, this is the first direct observation of intracellular transport of heme in a living eukaryotic cell. A similar approach, using Pd-mP fluorescence, could be applied to study heme intracellular metabolism in other cell types.

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A giant anteater (Myrmecophaga tridactyla) was found with closed comminuted fractures on the fight radius and ulna and left humerus he duration of which was unknown. The animal was unable to use either of he thoracic limbs. The fractures were stabilized with 3.5-mm titanium plates and a commercially available mixture of micro lyophilized bovine cortical osseous and bovine BMP (Gen-tech(R), Baumer, Brazil) was implanted into the fractures sites. Postoperative radiographic evaluations were performed every 30 days and after four months. Bone healing was observed in all of he fractures. The animal was able to be reintroduced into its natural habitat. From his case we conclude that despite he low metabolic fate of the giant anteater, which is an inherent characteristic of this species, he treatment of radius, ulna and humerus fractures by means of plates and screws, associated with BMP on the Myrmecophaga tridactyla, was a success.

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The fate of eight fungicides (benalaxyl, fenarimol, iprodione, metalaxyl, myclobutanil, procymidone, triadimefon, and vinclozolin) and five insecticides (dimethoate, fenthion, methidathion, parathion methyl, and quinalphos) in wine and its byproducts (cake and lees) during the production of distilled spirits was studied. Among the pesticides studied, only fenthion, quinalphos, and vinclozolin residues were present in the distilled spirits. During wine distillation, respectively 13% and 5% of the initial residues of fenthion and vinclozolin were transferred to the distilled spirit. Low percentages (2% for fenthion and 0.1% for vinclozolin) of these active ingredients (AI) also passed from the lees to the final-distilled spirit, when samples were fortified at 10.1 and 26.1 ppm for fenthion and vinclozolin, respectively. Quinalphos passed only from the lees to the final-distilled spirit in percentages lower than 1% when samples were fortified at the highest concentration (4.6 ppm).

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Over a 3-year period, all colony foundations of the social wasps Polistes versicolor and Polistes simillimus were registered, and the fate and growth of all colonies were followed. P. simillimus exhibited a greater number of colony-founding attempts, while P. versicolor had a larger number of adult colonies. P. simillimus had greater cell numbers and number of adults produced per colony. P. simillimus reutilized only a small percentage of brood cells for adult production for up to 2 generations, while P. versicolor reutilized a large percentage of brood cells for up to 3 generations. Consequently, cell production was higher in P. simillimus. Because of a high rate of adult production and extensive cell production, we suggest that P. simillimus may demonstrate paragynous social organization, and may demonstrate an intermediate form between polygynous and monogynous Vespidae. Paragynous associations may lead to lower predation pressures and a relative independence of adult production on time.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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The fate of folpet from the treatment on vine to the production of wine was studied. Sunlight degraded folpet to unknown products. Phthalimide was a minor metabolite formed on grapes from folpet. Folpet degraded in must, giving 80% phthalimide; the results obtained with model solutions showed that in must folpet can also give small amounts of phthalic acid. During wine-making folpet degraded completely, and at the end of fermentation phthalimide was only present in wine. This compound was stable in wine after several months. The presence of folpet in grapes inhibited the alcoholic fermentation of Saccharomyces cerevisiae and Kloeckera apiculata completely. Phthalimide, on the contrary, had no negative effect on the fermentative action of the two yeasts. GC and HPLC methods were developed to determine folpet and its metabolites.

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The persistence of three pesticides (fenitrothion, dimethoate, and ziram) in apricots in field conditions and their fate during the drying process were studied. After the treatments, the pesticides showed fast decay rates with pseudo-first-order kinetics and half-lives ranging from 6.9 to 9.9 days. The drying process showed a different effect on residue concentrations in dried apricots: omethoate (metabolite of dimethoate) and ziram residues had almost doubled, while fenitrothion disappeared and dimethoate remained constant.

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The effect of increasing phosphorus (P) intake on P utilization was investigated in balance experiments using 12 Saanen goats, 4 to 5 mo of age and weighing 20 to 30 kg. The goats were given similar diets with various concentrations of P, and 32P was injected to trace the movement of P in the body. A P metabolism model with four pools was developed to compute P exchanges in the system. The results showed that P absorption, bone resorption, and excretion of urinary P and endogenous and fecal P all play a part in the homeostatic control of P. Endogenous fecal output was positively correlated to P intake (P < .01). Bone resorption of P was not influenced by intake of P, and P recycling from tissues to the blood pool was lesser for low P intake. Endogenous P loss occurred even in animals fed an inadequate P diet, resulting in a negative P balance. The extrapolated minimum endogenous loss in feces was .067 g of P/d. The minimum P intake for maintenance in Saanen goats was calculated to be .61 g of P/ d or .055 g of P/(kg.75·d) at 25 kg BW. Model outputs indicate greater P flow from the blood pool to the gut and vice versa as P intake increased. Intake of P did not significantly affect P flow from bone and soft tissue to blood. The kinetic model and regressions could be used to estimate P requirement and the fate of P in goats and could also be extrapolated to both sheep and cattle.

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Due to the exclusively maternal inheritance of mitochondria, mitochondrial genotypes can be coupled to a particular nuclear genotype by continuous mating of founder females and their female offspring to males of the desired nuclear genotype. However, backcrossing is a gradual procedure that, apart from being lengthy, cannot ascertain that genetic and epigenetic changes will modify the original nuclear genotype. Animal cloning by nuclear transfer using host ooplasm carrying polymorphic mitochondrial genomes allows, among other biotechnology applications, the coupling of nuclear and mitochondrial genotypes of diverse origin within a single generation. Previous attempts to use Bos taurus oocytes as hosts to transfer nuclei from unrelated species led to the development to the blastocyst stage but none supported gestation to term. Our aim in this study was to determine whether B. taurus oocytes support development of nuclei from the closely related B. indicus cattle and to examine the fate of their mitochondrial genotypes throughout development. We show that indicus:taurus reconstructed oocytes develop to the blastocyst stage and produce live offspring after transfer to surrogate cows. We also demonstrate that, in reconstructed embryos, donor cell-derived mitochondria undergo a stringent genetic drift during early development leading, in most cases, to a reduction or complete elimination of B. indicus mtDNA. These results demonstrate that cross-subspecies animal cloning is a viable approach both for matching diverse nuclear and cytoplasmic genes to create novel breeds of cattle and for rescuing closely related endangered cattle.

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The HIV-infected individuals have been identified as a peculiar group whose propensity to the development of abnormalities in lipids metabolism supports the hypothesis that AIDS itself can be considered as an independent risk factor for the occlusive diseases development. The AIDS progression, as well as the therapy against HIV has been capable to show an array of metabolic disturbances that HIV-infected patients are prone to. These metabolic alterations affect the fate of plasmatic lipids and homocysteine as a result of three factor mainly: (i) the viral infection per se which triggers the development of hypertriglyceridemia and hipocholesterolemia; (ii) multiple vitamins and micronutrients deficiencies, that favors an onset of hyperhomocysteinemia; (iii) the state-of-the-art therapy for HIV infection, which is accompanied to idiosyncratic effects encompassing the lipid metabolism. In this context, a variety of risk factors to atherosclerosis can be identified in the HIV-infected individual. Of note, it must be considered that once life expectancy of these patients has been expanded due to the effective therapy, on the other hand they can accelerate atherosclerotic disease or its pathological appearance in the same extent.

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In order to assess a new strategy of DNA vaccine for a more complete understanding of its action in immune response, it is important to determine the in vivo biodistribution fate and antigen expression. In previous studies, our group focused on the prophylactic and therapeutic use of a plasmid DNA encoding the Mycobacterium leprae 65-kDa heat shock protein (Hsp65) and achieved an efficient immune response induction as well as protection against virulent M. tuberculosis challenge. In the present study, we examined in vivo tissue distribution of naked DNA-Hsp65 vaccine, the Hsp65 message, genome integration and methylation status of plasmid DNA. The DNA-Hsp65 was detectable in several tissue types, indicating that DNA-Hsp65 disseminates widely throughout the body. The biodistribution was dose-dependent. In contrast, RT-PCR detected the Hsp65 message for at least 15 days in muscle or liver tissue from immunized mice. We also analyzed the methylation status and integration of the injected plasmid DNA into the host cellular genome. The bacterial methylation pattern persisted for at least 6 months, indicating that the plasmid DNA-Hsp65 does not replicate in mammalian tissue, and Southern blot analysis showed that plasmid DNA was not integrated. These results have important implications for the use of DNA-Hsp65 vaccine in a clinical setting and open new perspectives for DNA vaccines and new considerations about the inoculation site and delivery system. © 2006 Coelho-Castelo et al; licensee BioMed Central Ltd.

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Purpose: Bone maintenance after mandibular reconstruction with autogenous iliac crest may be disappointing due to extensive resorption in the long term. The potential of the guided-bone regeneration (GBR) technique to enhance the healing process in segmental defects lacks comprehensive scientific documentation. This study aimed to investigate the influence of polylactide membrane permeability on the fate of iliac bone graft (BG) used to treat mandibular segmental defects. Materials and Methods: Unilateral 10-mm-wide segmental defects were created through the mandibles of 34 mongrel dogs. All defects were mechanically stabilized, and the animals were divided into 6 treatment groups: control, BG alone, microporous membrane (poly L/DL-lactide 80/20%) (Mi); Mi plus BG; microporous laser-perforated (15 cm2 ratio) membrane (Mip), and Mip plus BG. Calcein fluorochrome was injected intravenously at 3 months, and animal euthanasia was carried out at 6 months postoperatively. Results: Histomorphometry showed that BG protected by Mip was consistently related to larger amounts of bone compared with other groups (P ≤ .0001). No difference was found between defects treated with Mip alone and BG alone. Mi alone rendered the least bone area and reduced the amount of grafted bone to control levels. Data from bone labeling indicated that the bone formation process was incipient in the BG group at 3 months postoperatively regardless of whether or not it was covered by membrane. In contrast, GBR with Mip tended to enhance bone formation activity at 3 months. Conclusions: The use of Mip alone could be a useful alternative to BG. The combination of Mip membrane and BG efficiently delivered increased bone amounts in segmental defects compared with other treatment modalities. © 2008 American Association of Oral and Maxillofacial Surgeons.