Trichinella spiralis and breast carcinoma--a case report

Authors report about a patient with recurrent ductal invasive breast carcinoma and trichinosis. The patient underwent mastectomy of the left breast with evacuation of the axilla because of the cancer. Radiation therapy was received. An infestation with Trichinella spiralis was diagnosed two years after The patient was treated with mebendazole. A local recurrence of the tumor was found on the chest wall six years after the surgery. Tumor excision was performed. Histological analysis pointed at a ductal invasive carcinoma with numerous parasites of Trichinella spiralis present within both the muscle and the tumor tissue. The finding of parasites in the tumor tissue witnesses in favor of infestation, and the parasite morphology preserved in the tumor shows at the protective effects of the cysts, i.e. preventing parasite necrosis.     

<Emphasis Type="BoldItalic">Betula</Emphasis> spp. pollen in the atmosphere of Novi Sad (2000–2002)

Pollen of Betula spp. is one of the main European aeroallergens. The aim of this study was to determine characteristics and occurrence of the Betula pollen in Novi Sad atmosphere, based on 3-year observations (2000–2002), and to compare pollen season start dates calculated by different methods. Pollen samples have been collected by Hirst volumetric method with a 7-day Burkard spore trap. Four methods (Sum 75, 2.5%, 30 and 1 pg/m ³) have been used for determination of the start dates of the Betula pollen season and the results have been compared. The total annual pollen sum increased during the observed period. In 2000, 2001 and 2002, the highest daily pollen concentrations were 97, 137 and 1034 pg/m ³, respectively. The earliest Betula pollen season start has been calculated by the 1 pg/m ³ method.     

Xenopus laevis transgenesis by sperm nuclear injection

The stable integration of transgenes into embryos of the frog Xenopus laevis is achieved using the procedure described here. Linear DNA containing the transgene is incorporated randomly into sperm nuclei that have had their membranes disrupted with detergent treatment. Microinjection of these nuclei into unfertilized eggs produces viable embryos that can be screened for activity of the transgene. The proportion of embryos that harbor the transgene varies from 10 to 40% of the total number of surviving embryos. Multiple copies of the transgene can integrate as a concatemer into the sperm genome, and more than one site of DNA integration might occur within resulting animals. Germ cell transmission of the transgene is routine and the procedure is well suited to the production of transgenic reporter frog lines. One day should be allocated for the preparation of the sperm nuclei, which are stored as aliquots for future use. The transgenesis reaction and egg injection take one morning.     

Recent vegetation history of the North Adriatic grasslands: Expansion and decay of an anthropogenic habitat

A summary pollen diagram and old cartographic materials were used to reconstruct the recent vegetation history and trends of temporal dynamics of the North Adriatic Karst grasslands. Before the Roman period some evidence about grassland vegetation exists from Neolithic settlements, but deforestation, detectable on the landscape level because of pollen, started 2500-2000 years BP. A large-scale anthropogenically-driven process of clearance started in the Roman period as is reflected in the increasing pollen of grasses (Poaceae) and other herbaceous species (Apiaceae). The presence of pioneer trees (Quercus, Ostrya), juniper and grasses during 1000-400 years BP suggests that cycling processes of cultivation (strong grazing pressure), abandonment and re-cultivation were present. A 250-year-old map shows a peak of probable deforestation, where grasslands have 3.2 times larger surface area than on a recent land-use map in a 665.8 km² large pilot area. More than 60% of grasslands changed into forest since then. On a sub-plot area of 626 ha (1% of the pilot area), where habitat mappings were performed, only 12.8% were still grasslands without tall herb invasions or scrub encroachments. Another 27.1% were grasslands with early signs of reforestation. The traditionally open landscape of the North Adriatic Karst was able to recover to forest due to almost total abandonment and sufficient mesic climate conditions.     

A human origin type II strain of Toxoplasma gondii causing severe encephalitis in mice

Despite its capacity for sexual reproduction and global distribution, Toxoplasma gondii has a highly clonal structure, with the majority of isolates belonging to three distinct clonal types. Congenital toxoplasmosis has been associated with type I and type II strains. We here present the first characterization of a T. gondii strain (BGD1) from South-East Europe, isolated from the umbilical blood of a 24-week-old fetus in Serbia. Genotyping, performed by PCR-RFLP using a set of nested PCR markers (5'SAG2, 3'SAG2, BTUB, SAG3, and GRA6), showed that the BGD1 strain possessed a type II genotype. The cytokine patterns in Swiss-Webster mice inoculated with brain cysts of BGD1 and the prototype type II ME49 strain were similar until 180 days post-infection, with highly elevated IFN-gamma, IL-12 and IL-10 by d7 and decreasing thereafter. While both strains induced pneumonia and hepatitis in acute infection (d14), chronic infection (d56) was characterized, in addition to hepatitis, by severe meningoencephalitis, associated with numerous brain cysts. Thus, the BGD1 strain of T. gondii has type II genotypic and immunologic characteristics, but unlike other type II strains of human origin, induces severe encephalitis, making it an alternative to the sheep-derived ME49 strain for experimental models of infection.     

Developmental Expression Patterns of Arabidopsis XTH Genes Reported by Transgenes and Genevestigator

The plant cell wall is the structural basis of cellular form and thus forms a foundation on which morphogenesis builds organs and tissues. Enzymes capable of modifying major wall components are prominent candidates for regulating wall form and function. Xyloglucan endotransglucosylases/hydrolases (XTHs) are predicted to participate in xyloglucan integration and/or restructuring. XTHs are encoded by large gene families in plants; the Arabidopsis genome encodes 33 XTHs. To gain insight into the potential physiological relevance of the distinct members of this family, GUS reporter fusion genes were constructed, and plants expressing these transgenes were characterized to reveal spatial and temporal patterns of expression. In addition, Genevestigator sources were mined for comprehensive and comparative XTH expression regulation analysis. These data reveal that the Arabidopsis XTHs are likely expressed in every developmental stage from seed germination through flowering. All organs show XTH::GUS expression and most, if not all, are found to express multiple XTH::GUS genes. These data suggest that XTHs may contribute to morphogenesis at every developmental stage and in every plant organ. Different XTHs have remarkably diverse and distinct expression patterns indicating that paralogous genes have evolved differential expression regulation perhaps contributing to the maintenance of the large gene family. Extensive overlap in XTH expression patterns is evident; thus, XTHs may act combinatorially in determining wall properties of specific tissues or organs. Knowledge of gene-specific expression among family members yields evidence of where and when gene products may function and provides insights to guide rational approaches to investigate function through reverse genetics. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

Site-specific mutagenesis of the histidine precursor of diphthamide in the human elongation factor-2 gene confers resistance to diphtheria toxin

Protein synthesis elongation factor 2 (EF-2) from eukaryotes contains a conserved post-translationally modified histidine residue known as diphthamide. Diphthamide is a unique site of ADP-ribosylation by diphtheria toxin (DT), which is responsible for cell killing. In this report, we describe the construction of DT-resistant HeLa cell lines by engineering the toxin-resistant form of its specific substrate, protein elongation factor-2. Using site-specific mutagenesis of the histidine precursor of diphthamide, the histidine residue of codon 715 in human EF-2 cDNA was substituted with one of four amino acid residue codons: leucine, methionine, asparagine or glutamine. Mutant EF-2s were subcloned into a pCMVexSVneo expression vector, transfected into HeLa cells, and DT-resistant cell clones were isolated. The protective effect of mutant EF-2s against cell killing by DT, after exposing all four mutant strains derived from HeLa cells to different concentrations of the toxin (5-20 ng/mL) was demonstrated by: (1) the normal morphological appearance of the cells; (2) their unaffected or slightly slower growth rates; (3) their undisturbed electrophoretic DNA profiles whose integrity was virtually preserved. Mutant cell strains showed also considerable levels of resistance to very high concentrations of DT, in that they maintained slower but consistent rates of cell growth. It was hence concluded that despite its strict conservation and unique modification, the diphthamide histidine appears not to be essential to the function of human EF-2 in protein synthesis. In addition, DT-resistant HeLa cell clones should prove valuable hosts for various DT gene-containing vectors that express the toxin intracellularly.