An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry

Characterization of host-pathogen interactions is a fundamental approach in microbiological and immunological oriented disciplines. It is commonly accepted that host cells start to change their phenotype after engulfing pathogens. Techniques such as real time PCR or ELISA were used to characterize the genes encoding proteins that are associated either with pathogen elimination or immune escape mechanisms. Most of such studies were performed in vitro using primary host cells or cell lines. Consequently, the data generated with such approaches reflect the global RNA expression or protein amount recovered from all cells in culture. This is justified when all host cells harbor an equal amount of pathogens under experimental conditions. However, the uptake of pathogens by phagocytic cells is not synchronized. Consequently, there are host cells incorporating different amounts of pathogens that might result in distinct pathogen-induced protein biosynthesis. Therefore, we established a technique able to detect and quantify the number of pathogens in the corresponding host cells using immunofluorescence-based high throughput analysis. Paired with multicolor staining of molecules of interest it is now possible to analyze the infection profile of host cell populations and the corresponding phenotype of the host cells as a result of parasite load.     

Transmission of parasites in the coastal waters of the Arctic seas and possible effect of climate change

The review deals with the biodiversity, life cycles, distribution and temperature adaptations of parasites circulating in the coastal waters of northern polar seas. Special attention is given to helminths of marine birds, which are the most common parasites in the coastal waters. Among them, the focus is on trematodes. Factors responsible for the impoverished species composition of parasites in the Arctic are analyzed. It is shown that species without free-living larvae in the life cycle have an advantage in this environment. The abundance of cestodes and acanthocephalans in Arctic seabirds is linked with the high proportion of crustaceans in their diet. The phenomenon of nonspecific parasitism (occurrence of parasites in atypical host species) is analyzed from an evolutionary viewpoint. Characteristic features in the spatial distribution of infection of marine coastal invertebrates with parasite larvae are considered, and factors that determine it are specified. The prevalence of infection in intermediate hosts is closely connected with the abundance of final hosts, which makes it possible to estimate the abundance of final hosts in a given region and reveal trends in its changes. Trematodes have a high potential for temperature acclimation. This facilitates their transmission in the northern seas but, on the other hand, makes it unlikely that the transmission process would be intensified upon an increase in summer temperatures resulting from climate warming. However, intensification of transmission may well occur due to the prolongation of the warm season (“transmission window”), which has been predicted and is already observed. It is suggested that warming in the Arctic promotes both the entry of certain “southern” species into the Arctic and the trans-Arctic interpenetration of the North Atlantic and North Pacific parasitic faunas. A case is made for the necessity to broaden the scope of parasitological research in the Arctic and Subarctic, including parasitological monitoring at the reference sites of the sea coast.     

The Cultural-Historical Development of Mental Processes and the Theory of Stepwise Formation of Mental Actions and Concepts

The article examines the problem of the relationship between universal and specific forms of cultural–historical development of mental processes. From different methodological approaches, the author prefers an activity approach which emphasizes genetic primacy of external practical activity. This approach allows, according to the author, universality of internalization and at the same time ethnic uniqueness of mental processes. The article clarifies the problem of internalization through two practical examples: 1) the development of mental counting actions and 2) the process of human goal setting.     

Mycelial elongation and sporulation of two fungi on amended media in light or dark

Botrytis allii andCollectotrichum dematium are onion pathogens which can infect in the field and cause decay in storage. Some phenolics can hinder development of these fungi, but the effect of cytokinins is not clear. Cytokinins (kinetin or 6-benzyladenine) or phenolics (caffeic or chlorogenic acids) were added to agar at concentrations of 0 to 10^–3 M. Cultures were continuously irradiated with fluorescent light or maintained in the dark for 6 days. On unamended media, final mycelial elongation was 45 or 17.8 mm and sporulation was 28 or 10.6 × 10⁴ spores/ml forBotrytis andColletotrichum, respectively. ForBotrytis, mycelial elongation was slightly (5%) but significantly increased and sporulation increased by 21% by incubation on phenolics as compared to cytokinins. Mycelial extension ofColletotrichum was not affected by amendment. Sporulation ofColletotrichum on kinetin was 16 to 28% greater than on the other amendments. As amendments concentration increased elongation of mycelia of both fungi decreased. Sporulation ofBotrytis increased by 60% as amendment concentration increased from 0 to 10^–5 M and then decreased 25% at 10^–3 M. As amendment concentration increased from 0 to 10^–3 M, sporulation ofColletotrichum increased by 45%. Incubation in light increased mycelial extension 3 to 17% forBotrytis andColletotrichum respectively, and sporulation was increased approximately 78% for both fungi. These compounds do not appear to inhibit development of theseBotrytis orColletotrichum species in culture.