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The aim of the study was to enhance specificity of immunological diagnosis in toxocarosis. The investigations concem: detection of circulating parasite antigens using dot-ELISA technique and detection of specific IgG antigodies using ELISA test with low molecular weight (≤30 kD) excretory-secretory (ES) larval T. canis antigens. The circulating antigens were investigated in 60 sera and 20 anterior chamber fluids from children with ocular form of toxocarosis. These antigens were present only in 3 sera and 2 anterior chamber fluids in children with active intraocular inflamation and fluid antibodies titers 1:5120 and 1:20000. Comparative study with different ELISA tests indicated that use of low molecular weight antigens increase specificity of reaction.
The dynamics of IgA- and IgG-containing cells in the stomach mucosa of rabbits infected with Obeliscoides cuniculi was studied by means of a direct immunofluorescence method. An increase in the numbers of IgA-containing cells was found during the histotrophic phase of infection. An aggregation of secretory IgA on the external surface of the cuticle of worms was also observed. There was no stimulation of proliferation of local IgG-containing cells. The numbers of IgG-containing cells in infected rabbits were very low, and also very few IgG-containing cells were found in the stomach mucosa of uninfected rabbits. A weak immunological response in the blood sera, was observed only in a few animals after infection with various doses of larvae. No difference was found in the level of antibody titers as a result of exposure of animals to various doses of larvae. A significant increase in the number of IgA-containing cells in the stomach of infected rabbits might imply that О. cuniculi stimulated a local immune response of parasitized rabbits.
Przedstawiono opis procedur zaadaptowanych do izolacji i wykrywania DNA pasożytów z rodzaju Dirofilaria w materiale izolowanym od ludzi i przechowywanym w: etanolu (24 tygodnie), formalinie (46 tygodni) lub zatopionym w bloczkach parafinowych (25 tygodni). Najwyższą czułość PCR obserwowano dla próbek DNA uzyskanych z robaków przechowywanych w etanolu, a najniższą dla próbek pochodzących z materiału przechowywanego w formalinie. Izolacja DNA z pasożyta utrwalonego w formalinie była trudniejsza niż w wypadku pozostałych preparatów, a produkty jego powielania były wykrywalne tylko w Real Time PCR. Uzyskane wyniki pokazują, że metody molekularne są stosunkowo łatwym w użyciu sposobem identyfikacji nicieni z rodzaju Dirofilaria obecnych w materiale klinicznym utrwalonym na różne sposoby. Wskazane jest niestosowanie formaliny do przechowywania materiału, który ma być użyty do diagnostyki opartej o PCR.
The identified mutations in the pfcrt, dhfr and dhps genes of Plasmodium falciparum show a very high correlation with resistance to chloroquine, pyrimethamine and sulfadoxine, the drugs that are still used as malaria chemoprophylaxis or treatment. We undertook a molecular screening of 82 Polish P. falciparum isolates, mainly imported from different countries of sub-Saharan Africa to assess their molecular drug-resistance profiles. Only 4 isolates showed no mutations in the three analyzed gene fragments. In the remaining isolates from one to six mutations in one or more examined genes were found. Different mutations in the pfcrt, dhfr and dhps genes were found in ca. 76%, 80% and 70% of P. falciparum isolates, respectively. About forty our patients used chloroquine or pyrimethamine + sulfadoxine as malaria chemoprophylaxis and/or antimalarial treatment, but without success. In all but 5 of the P. falciparum isolates obtained from these persons, mutations associated to resistance of the parasite to chloroquine and the antifolate drugs were found.
Dirofilaria (Nochtiella) repens Railliet et Henry, 1911 (Nematoda: Onchocercidae) is a subcutaneous parasite of dogs and other carnivorous animals, with human acting as incidental hosts. D. repens occurs endemically in warm climates on various continents, in Europe mainly in Mediterranean countries. The aim of this study was to summarize information on human dirofilariosis in Poland, taking into consideration parasitological and epidemiological data. Between April 2009 – December 2011, in the parasitological laboratories of Medical University in Warsaw and the National Institute of Public Health/National Institute of Hygiene, fragments of affected human tissues and parasite specimens were examined microscopically. Molecular methods were used to confirm the results from eight microscopic investigations. A literature review to summarize all data on dirofilarial infections in humans in Poland was conducted. In these investigations, autochthonous dirofilariosis was found in humans for the first time in Poland. During the last 3 years, 12 new cases of human D. repens dirofilariosis were recognized. Since 2007, a total of 18 D. repens infection have been found in humans in Poland. Parasitic changes were located in various parts of the body, in the form of subcutaneous nodules containing single nematodes surrounded by granulation tissue (15 cases). In 3 cases, a subconjuctival localization was found. Seventeen of the 18 described cases were noted in central Poland where dirofilariosis occured in dogs. In this area, autochtonous infection was identified in 3 women who had never left Poland in their lives; the others were probably infected outside the country while staying in endemic regions. Data on human and canine infection collected from central Poland during the last 5 years indicates that Dirofilaria repens has been introduced into our country, and that the infection is successfully spreading, with the border of the endemic area currently on 52°N, 21°E. To control the epidemiological situation it is necessary to identify D. repens hosts within local mosquito populations, and to monitor dogs. Because of the increasing number of cases of human infections, whether introduced or local, physicians should take dirofilariosis into consideration in differential diagnosis of skin and eye diseases.
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