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1.
Calvet CM Toma L De Souza FR Meirelles Mde N Pereira MC 《The Journal of eukaryotic microbiology》2003,50(2):97-103
Cytoadherence is an important step for the invasion of a mammalian host cell by Trypanosoma cruzi. Cell surface macromolecules are implicated in the T. cruzi-cardiomyocyte recognition process. Therefore, we investigated the role of cell surface proteoglycans during this invasion process and analyzed their expression after the parasite infected the target cells. Treatment of trypomastigote forms of T. cruzi with soluble heparan sulfate resulted in a significant inhibition in successful invasion, while chondroitin sulfate had no effect. Removal of sulfated glycoconjugates from the cardiomyocyte surface using glycosaminoglycan (GAG) lyases demonstrated the specific binding of the parasites to heparan sulfate proteoglycans. Infection levels were reduced by 42% whenthe host cells were previously treated with heparitinase II. No changes were detected in the expression of GAGs infected cardiomyocytes even after 96 h of infection. Our data demonstrate that heparan sulfate proteoglycans, but not chondroitin sulfate, mediate both attachment and invasion of cardiomyocytes by T. cruzi. 相似文献
2.
da Silveira RB Pigozzo RB Chaim OM Appel MH Silva DT Dreyfuss JL Toma L Dietrich CP Nader HB Veiga SS Gremski W 《Biochimie》2007,89(3):289-300
Loxoscelism (the condition produced by the bite of brown spiders) has been reported worldwide, but especially in warmer regions. Clinical manifestations include skin necrosis with gravitational spreading while systemic loxoscelism may include renal failure, hemolysis and thrombocytopenia. The venom contains several toxins, of which the best biochemically and biologically studied is the dermonecrotic toxin, a phospholipase-D. Purified toxin induces cutaneous and systemic loxoscelism, especially necrotic lesions, hematological disturbances and renal failure. Herein, we describe cloning, heterologous expression and purification of two novel dermonecrotic toxins: LiRecDT4 and LiRecDT5. The recombinant proteins stably expressed in Escherichia coli cells were purified from culture supernatants in a single step using Ni(2+)-chelating chromatography producing soluble proteins of 34 kDa (LiRecDT4) and 37 kDa (LiRecDT5). Circular dichroism analysis evidenced correctly folding for toxins but differences in secondary structures. Both proteins were recognized by whole venom serum antibodies and by a specific antibody to dermonecrotic toxin. Also, recombinant toxins with phospholipase activity induced experimental skin lesions and caused a massive inflammatory response in rabbit skin dermis. Nevertheless, toxins displayed different effects upon platelet aggregation, increase in vascular permeability and not caused death in mice. These characteristics in combination with functional studies illustrates that a family of dermonecrotic toxins exists, and includes two novel members that are useful for future structural and functional studies. They will also be useful in biotechnological ends, for example, as inflammatory and platelet aggregating studies, as antigens for serum therapy source and for lipids biochemical research. 相似文献
3.
Andrew J.?NossEmail author Imke?Oetting Rosa?Leny?Cuéllar 《Biodiversity and Conservation》2005,14(11):2679-2693
Declared in 1995, the 34,400 km2 Kaa-Iya del Gran Chaco National Park is the first protected area in South America co-managed by an indigenous organization, the Capitanía del Alto y Bajo Isoso (CABI). In 1997, based on historical occupation by the Isoseño-Guaraní over the past 300 years, CABI formally demanded a 19,000 km2 `Tierra Comunitaria de Orígen' (TCO) that adjoins, but does not overlap, the national park. The creation of TCOs and the co-administration of protected areas are elements of decentralization processes in Bolivia, whereby the management of land and natural resources is devolving to departmental and municipal levels of government. This paper examines biodiversity monitoring in the context of a community wildlife management program developed with CABI. Hunter self-monitoring (100–150 hunters per month) combined with monthly activity records for potential hunters (7637 observed hunter-months) permit estimations of total offtakes of subsistence game species for 1996–2003, as well as catch-per-unit-effort over the same time period. These data show considerable fluctuations from year to year and no declining trends that would suggest over-hunting. Monitoring populations of multiple game species can be relatively expensive, even with the voluntary support of hunters, considering data collection and analysis, as well as presentation and discussion through community meetings. At the same time, monitoring does not provide highly accurate assessments of short-term changes in wildlife resources. However, relatively simple participatory methods are important for generating information on long-term trends and for creating a context for community discussion of formal wildlife management. 相似文献
4.
Evolutionary dynamics of the kinetochore network in eukaryotes as revealed by comparative genomics
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Jolien JE van Hooff Eelco Tromer Leny M van Wijk Berend Snel Geert JPL Kops 《EMBO reports》2017,18(9):1559-1571
During eukaryotic cell division, the sister chromatids of duplicated chromosomes are pulled apart by microtubules, which connect via kinetochores. The kinetochore is a multiprotein structure that links centromeres to microtubules, and that emits molecular signals in order to safeguard the equal distribution of duplicated chromosomes over daughter cells. Although microtubule‐mediated chromosome segregation is evolutionary conserved, kinetochore compositions seem to have diverged. To systematically inventory kinetochore diversity and to reconstruct its evolution, we determined orthologs of 70 kinetochore proteins in 90 phylogenetically diverse eukaryotes. The resulting ortholog sets imply that the last eukaryotic common ancestor (LECA) possessed a complex kinetochore and highlight that current‐day kinetochores differ substantially. These kinetochores diverged through gene loss, duplication, and, less frequently, invention and displacement. Various kinetochore components co‐evolved with one another, albeit in different manners. These co‐evolutionary patterns improve our understanding of kinetochore function and evolution, which we illustrated with the RZZ complex, TRIP13, the MCC, and some nuclear pore proteins. The extensive diversity of kinetochore compositions in eukaryotes poses numerous questions regarding evolutionary flexibility of essential cellular functions. 相似文献
5.
Coulson-Thomas VJ Coulson-Thomas YM Gesteira TF de Paula CA Mader AM Waisberg J Pinhal MA Friedl A Toma L Nader HB 《Cell and tissue research》2011,346(2):223-236
During cancer cell growth many tumors exhibit various grades of desmoplasia, unorganized production of fibrous or connective
tissue, composed mainly of collagen fibers and myofibroblasts. The accumulation of an extracellular matrix (ECM) surrounding
tumors directly affects cancer cell proliferation, migration and spread; therefore the study of desmoplasia is of vital importance.
Stromal fibroblasts surrounding tumors are activated to myofibroblasts and become the primary producers of ECM during desmoplasia.
The composition, density and organization of this ECM accumulation play a major role on the influence desmoplasia has upon
tumor cells. In this study, we analyzed desmoplasia in vivo in human colorectal carcinoma tissue, detecting an up-regulation of collagen I, collagen IV and collagen V in human colorectal
cancer desmoplastic reaction. These components were then analyzed in vitro co-cultivating colorectal cancer cells (Caco-2 and HCT116) and fibroblasts utilizing various co-culture techniques. Our findings
demonstrate that direct cell-cell contact between fibroblasts and colorectal cancer cells evokes an increase in ECM density,
composed of unorganized collagens (I, III, IV and V) and proteoglycans (biglycan, fibromodulin, perlecan and versican). The
desmoplastic collagen fibers were thick, with an altered orientation, as well as deposited as bundles. This increased ECM
density inhibited the migration and invasion of the colorectal tumor cells in both 2D and 3D co-culture systems. Therefore
this study sheds light on a possible restricting role desmoplasia could play in colorectal cancer invasion. 相似文献
6.
Hendrik G. de Vries J. Margriet Collée Marjan H. R. van Veldhuizen Leny Achterhof Cees Th. Smit Sibinga Hans Scheffer Charles H. C. M. Buys Leo P. ten Kate 《Human genetics》1996,97(3):334-336
Mouthwashes can be used as a DNA resource for mutation detection and, because collection and DNA isolation is simple and cheap, they could in particular, be used for large numbers of samples. To determine the failure rate (the proportion of mouth samples in which no PCR product was obtained) and the specificity of buccal epithelial cell mutation detection in large numbers of samples, we collected mouthwashes and blood samples from 11413 blood donors and tested the mouthwashes for the F508 mutation, which has an estimated frequency of 75% among cystic fibrosis chromosomes in The Netherlands. Blood samples were tested for the F508 mutations only if the mutation was identified in the mouthwash or in the case of a failure to obtain PCR products. The sensitivity of the test was determined in mouthwashes of 75 F508 carriers known from earlier family studies. These samples were offered blindly between the mouthwashes of the blood donors. Both specificity and sensitivity of the mouthwash procedure were 100%. The overall failure rate was 5.6%. This large figure was caused mainly by insufficient rinsing of the mouth in one particular blood bank. Exclusion of the results of this blood bank reduced the failure rate to 1.8%. Our results also confirm that for a large number of samples the mouthwash procedure is suitable for mutation detection and, with proper instructions, can be used in community screening. 相似文献
7.
Andrew?J.?NossEmail author Erika?Cuéllar Rosa?Leny?Cuéllar 《Human ecology: an interdisciplinary journal》2004,32(6):685-702
Community wildlife management is being tested across the tropics as a means of promoting the conservation and sustainable use of wildlife resources. Key to successful programs is the effective participation of local hunters and communities in monitoring, planning, decision-making and implementation. We evaluate one method to achieve this participation, namely hunter self-monitoring. Between 1997 and 2000, Izoceño hunters from 22 communities in the Bolivian Chaco have voluntarily participated in monitoring their hunting activities, measuring and recording data on captured animals and hunting methods in personal notebooks. Despite the lack of remuneration, participation exceeds 60% of active hunters. However, the written information and specimens provided are not complete, and are biased according to hunting methods and prey characteristics. Complementary research is essential to answer specific research questions. Nevertheless, hunter self-monitoring serves to raise awareness of wildlife management issues at the communal or indigenous territory level, as evidenced by preliminary actions taken by hunters and communities in the Izozog. 相似文献
8.
Victoria R. Rendell Robert H. Gilman Edward Valencia Gerson Galdos-Cardenas Manuela Verastegui Leny Sanchez Janet Acosta Gerardo Sanchez Lisbeth Ferrufino Carlos LaFuente Maria del Carmen Abastoflor Rony Colanzi Caryn Bern 《PloS one》2015,10(3)
Background
Congenital transmission is a major source of new Trypanosoma cruzi infections, and as vector and blood bank control continue to improve, the proportion due to congenital infection will grow. A major unanswered question is why reported transmission rates from T. cruzi-infected mothers vary so widely among study populations. Women with high parasite loads during pregnancy are more likely to transmit to their infants, but the factors that govern maternal parasite load are largely unknown. Better understanding of these factors could enable prioritization of screening programs to target women most at risk of transmission to their infants.Methodology/Principal Findings
We screened pregnant women presenting for delivery in a large urban hospital in Bolivia and followed infants of infected women for congenital Chagas disease. Of 596 women screened, 128 (21.5%) had confirmed T. cruzi infection; transmission occurred from 15 (11.7%) infected women to their infants. Parasite loads were significantly higher among women who transmitted compared to those who did not. Congenital transmission occurred from 31.3% (9/29), 15.4% (4/26) and 0% (0/62) of women with high, moderate and low parasite load, respectively (χx2 for trend 18.2; p<0.0001). Twin births were associated with higher transmission risk and higher maternal parasite loads. Infected women without reported vector exposure had significantly higher parasite loads than those who had lived in an infested house (median 26.4 vs 0 parasites/mL; p<0.001) with an inverse relationship between years of living in an infested house and parasite load.Conclusions/Significance
We hypothesize that sustained vector-borne parasite exposure and repeated superinfection by T. cruzi may act as an immune booster, allowing women to maintain effective control of the parasite despite the down-regulation of late pregnancy. 相似文献9.
Litia A. Carvalho Louise C. Vitorino Roberta P.M. Guimarães Silvana Allodi Ricardo A. de Melo Reis Leny A. Cavalcante 《Biochemical and biophysical research communications》2014
We examined the effects of conditioned medium from olfactory ensheathing glia (OEGCM) on the differentiation of oligodendrocytes in mixed cultures of early postnatal hippocampi. Differentiation was judged from the numerical density (ND) of cells immunoreactive to 2′3′ cyclic nucleotide 3′phosphodiesterase (CNPase) and O4 antibodies. NDs increased according to inverted-U dose–response curves, particularly for CNPase+ cells (9-fold at optimal dilution) and these changes were blocked by inhibitors of ERK1, p38-MAPK, and PI3K. Our results raise the possibility that OEG secreted factor(s) may counteract demyelination induced by trauma, neurodegenerative diseases, and advanced age, and should stimulate novel methods to deliver these factors and/or potentiating chemicals. 相似文献
10.
Wagner Baetas-da-Cruz Roger M Macedo-Silva Alessandra Santos-Silva Andrea Henriques-Pons Maria F Madeira Suzana Corte-Real Leny A Cavalcante 《The journal of histochemistry and cytochemistry》2004,52(8):1047-1055
Leishmania amazonensis, an obligatory intracellular parasite, survives internalization by macrophages, but no information is available on the involvement of microglia. We have investigated microglia-protozoa interactions in mixed glial cultures infected with promastigote forms of L. amazonensis after lipopolysaccharide (LPS) or dexamethasone (DM) treatment. After 2 hr of exposure to parasites in control cultures, there was a small number of infected microglia (1%). Preincubation with LPS or DM led to 14% or 60% of microglial cells with attached parasites, respectively. DM treatment resulted in 39% of microglial cells with internalized parasites (controls or LPS-treated cells had < or =1%). Scanning electron micrographs showed numerous filopodia in DM-treated cells, whereas these projections were rarely observed in LPS-treated or control cells. DM treatment also affected the intramicroglial survival of Leishmania. In control cultures, internalized parasites, tagged with an anti-lipophosphoglycan (anti-LPG) antibody, showed fragmented DNA [terminal deoxyribonucleotide transferase-mediated dUTP-X nick end labeling (TUNEL+)] after 4 hr of interaction, but changes seemed slightly delayed in DM-treated cultures. After 12 hr, there were no LPG+/TUNEL+ profiles in controls, whereas rare LPG+ profiles still persisted in DM-treated cells. Our results suggest that microglia are highly effective in the elimination of Leishmania and that the process can be effectively studied by LPG/TUNEL double labeling. 相似文献