Signaling life and death in the thymus: timing is everything

T lymphocytes are generated in the thymus, where developing thymocytes must accept one of two fates: They either differentiate or they die. These fates are chiefly determined by signals that originate from the T cell receptor (TCR), a single receptor complex with a remarkable capacity to decide between distinct cell fates. This review explores TCR signaling in thymocytes and focuses on the kinetic aspects of ligand binding, coreceptor involvement, protein phosphorylation, and mitogen-activated protein kinase (MAPK) activation. Understanding the logic of TCR signaling may eventually explain how thymocytes and T cells distinguish self from nonself, a phenomenon that has fascinated immunologists for 50 years.     

Crystallography of the hexagonal ferrites

The hexagonal ferrites form an unusual group of complex, ferrimagnetic oxides embodying some 60 known crystal structures. These include phases for which the structural unit cell is larger than that in any known inorganic materials. The various hexagonal ferrite modifications fall into two distinct structural series, each formed by the ordered interlayering (stacking) of two discrete building blocks; these blocks stack along the c crystallographic axis in varying ratios and varying permutations to form strictly coherent, reproducible crystal structures. This mixed-layering aspect of the hexagonal ferrites permits direct, visual observation of the sequence of their subunit-cell stacking elements, after etching, by means of electron microscopy. The sequence of stacked blocks in such structures constitutes the only information lacking for a complete, three-dimensional structure determination. Direct access to this information provides an immediate, unique solution of the crystal structure problem in each case and thereby avoids the dilemmas of a classical diffraction approach to such large unit cells. Ferrite structures with hexagonal c dimensions of 1455 and 1577 angstroms have been uniquely solved by direct electron microscopic readout of surface etch features. One must exercise caution, however, in generalizing these findings to other materials. The method is successful in the case of the hexagonal ferrites because these are mixed-layer structures, wherein the building blocks react at different rates to a specific etchant. Mixed-layer systems are not uncommon in crystallography, and it is likely that similar techniques can be developed for other such materials. Regardless of the validity of this prognosis, however, it is quite evident that high-resolution replica electron microscopy is a most promising tool for the direct observation of surface structure on an ultramicro scale. During the studies reported here replica resolution capability was improved to about 10 angstroms; final resolution is limited by the particle size of the platinum shadowing material. Careful control of experimental conditions during replica preparation or an alternate choice of shadowing material, or both, might reasonably improve the resolution by a factor of 2. This resolution is within the range of most unit cell dimensions and approaches interatomic distances in solid-state materials. The potential of such an experimental capability needs no elaboration.     

G protein signalling involved in host recognition and mycoparasitismrelated chitinase expression in Trichoderma atroviride

Mycoparasitic species of Trichoderma are commercially applied as biological control agents against various fungal pathogens. The mycoparasitic interaction is host specific and includes recognition, attack and subsequent penetration and killing of the host. Investigations on the underlying events revealed that Trichoderma responds to multiple signals from the host (e.g. lectins or other ligands such as low molecular weight components released from the host's cell wall) and host attack is ac…     

Stable isotope N and C labelling of different functional groups of earthworms and their casts: A tool for studying trophic links

Earthworms (Oligochaeta: Lumbricidae) have substantial effects on the structure and fertility of soils with consequences for the diversity of plant communities and associated ecosystem functions. However, we still lack a clear understanding of the functional role earthworms play in terrestrial ecosystems, partly because easy-to-use methods to quantify their activities are missing. In this study, we tested whether earthworms and their casts can be dual-labelled with ¹⁵N and ¹³C stable isotopes by cultivating them in soil substrate amended with ¹⁵N ammonium nitrate and ¹³C-glucose. Additionally, we also wanted to know whether (i) earthworms from different functional groups (soil-feeders vs. litter-feeders) and their casts would differ in their incorporation of stable isotopes, (ii) if enrichment levels are higher if the same amount of isotopes is applied in one dose or in staggered doses, and (iii) if isotopic enrichment in casts changes when they are stored in a conditioning cabinet or in a pot filled with soil placed in a greenhouse. Our findings show the feasibility of dual-labelling tissues and casts of both litter-feeding (Lumbricus terrestris) and soil-feeding (Aporrectodea caliginosa) earthworms using the same method. The advantage of this method is that earthworms and their casts can be labelled under realistic conditions by cultivating them for only four days in soil that received a one-time addition of commercially available stable isotopes instead of offering labelled plant material. In earthworms, the isotopic enrichment remained at a stable level for at least 21 days; labelled casts could be stored for at least 105 days without significantly decreasing their isotopic signals. This simple and efficient method opens new avenues for studying the role of these important ecosystem engineers in nutrient cycling and their functional relationships with other organisms.     

Immunochemical and mass spectrometry detection of residual proteins in gluten fined red wine

Recently, wheat gluten has been proposed as technological adjuvant in order to clarify wines. However, the possibility that residual gluten proteins remain in treated wines cannot be excluded, representing a hazard for wheat allergic or celiac disease patients. In this work, commercial wheat glutens, in both partially hydrolyzed (GBS-P51) and nonhydrolyzed (Gluvital 21000) forms, were used as fining agents in red wine at different concentrations. Beside immunoenzymatic analyses using anti-gliadin, anti-prolamin antibodies and pooled sera of wheat allergic patients, a method based on liquid chromatography coupled to mass spectrometry has been proposed to detect residues of gluten proteins. Residual gluten proteins were detected by anti-prolamin antibodies, anti-gliadin antibodies and sera-IgE only in the wine treated with GBS-P51 at concentration 50, 150, and 300 g/hL, respectively, whereas no residual proteins were detected by these systems in the wine treated with Gluvital 21000. In contrast liquid chromatography-mass spectrometry analyses allowed the detection of proteins in red wines fined down to 1 g/hL of Gluvital 21000 and GBS-P51. Our results indicate that MS methods are superior to immunochemical methods in detecting gluten proteins in wines and that adverse reactions against gluten treated wines cannot be excluded.     

Species-specific sensitivity of earthworm coelomocytes to dermal metal (Cd,Cu, Ni,Pb, Zn) exposures: Methodological approach

Earthworm immune-competent cells (coelomocytes) can be adversely affected by soil metal contamination. The aim of the present paper was to perform comparative studies on the coelomocytes of four lumbricid species dermally exposed for 2 days to filter paper soaked with Ni, Cu, Zn, Cd, or Pb chlorides (1 mM). Coelomocytes were expelled by electro-stimulation, counted, and assayed ex vivo by flow cytometry and spectrofluorometry. The coelomocytes of Allolobophora chlorotica and Dendrobaena veneta contain both amoebocytes and riboflavin-storing eleocytes; consequently, they were subjected to assays based on flow cytometric quantification of autofluorescent eleocytes and spectrofluorometric measurement of riboflavin content in coelomocyte lysates. In Al. chlorotica, the number of coelomocytes, the percentage of eleocytes, and the amount of riboflavin were significantly lower in Cu-exposed worms although these cytometric parameters were less affected by Ni, Zn, Cd exposure, and entirely unaffected by Pb exposure. In D. veneta, such cellular effects were also only observed in Cu-exposed worms. The coelomic fluids of Aporrectodea caliginosa and Lumbricus rubellus are constitutionally impoverished of eleocytes; therefore, these species could not be subjected to assays measuring the riboflavin content of these granular cells. Rather, the amoebocytes of Ap. caliginosa and L. rubellus were subjected to flow cytometric measurements of in vitro neutral red uptake (NR). However, the NR uptake assay was technically demanding, requiring a strictly normalized incubation period over all samples to yield useful comparative data. In contrast, the riboflavin content in the coelomocyte lysates of eleocyte-rich species appears to be a robust and convenient immune-function biomarker of environmental stress.     

Riboflavin storage in earthworm chloragocytes and chloragocyte-derived eleocytes and its putative role as chemoattractant for immunocompetent cells

We have shown previously that riboflavin is stored in chloragosomes of chloragocyte-derived earthworm eleocytes what may have an adaptive value for worms vulnerable to soil-derived pathogen invasion. Thus, it is important to determine whether riboflavin stores are also present in earthworm species with low eleocyte counts, especially in the chloragocytes of their chloragogen tissue that surrounds the coelomic site of the intestine, and find out its putative roles in earthworm immunity. Experiments were performed on adult worms from 6 species. Freely floating coelomocytes (consisting of amoebocytes plus species-specific numbers of eleocytes) were extruded by mild electric shock and then chloragocytes from the same anaesthetized worm were mechanically detached into buffer. Both cell suspensions were analyzed by spectrofluorometry for riboflavin content. It turned out that riboflavin is stored in species-specific quantities in all investigated species. Riboflavin storage predominates in free coelomocytes of eleocyte-rich species (Eisenia andrei, Dendrobaena veneta, Allolobophora chlorotica) while in chloragocytes of species with few eleocytes (Lumbricus terrestris, L. rubellus, and Aporrectodea caliginosa). Upon a massive microbial impact, the coelomocytes (both amoebocytes and eleocytes) are involved in the formation of multicellular bodies encapsulating soil-derived pathogens, what is connected with enhanced coelomocyte mobility. Thus the second aim of the present investigation was to check if riboflavin can participate in coelomocyte accumulations. Tests performed in a 48-well chemotaxis chamber revealed that riboflavin behave as chemoattractant for coelomocytes of all investigated earthworm species, which may be one of mechanisms underpinning its immunomodulatory functions.     

Relating genetic variation of ecologically important tree traits to associated organisms in full-sib aspen families

Knowledge on phenological, morphometric, and phytochemical variation of local progenies of European aspen (Populus tremula, L.) is limited. The goal of this study was to characterize variation in growth and ecologically important leaf properties in aspen full-sib families in relation to interacting organisms (mycorrhiza, endophytes, and insects) and to determine whether these interactions were affected by soil application of a systemic fungicide. In local progenies, within-family variation of neutral molecular genetic markers (nuclear microsatellites) was higher than between families. Significant variation in growth, production of phenolic defensive compounds and other phytochemical leaf traits was found between families. Phenolic compounds showed clear negative correlation with generalist herbivores, but did not result in negative trade-off with biomass production. Differences in mycorrhizal colonization were not found among full-sib families and application of a systemic fungicide suppressed neither mycorrhizal colonization nor infestation with insects. However, a strong suppression of endophytes occurred, whose long-term consequences may require attention when fungicides are used in agroforestry plantations.     

Abdominal irradiation as a rescue therapy for feline gastrointestinal lymphoma: a retrospective study of 11 cats (2001-2008)

In this retrospective study medical records of 11 cats with gastrointestinal lymphoma were evaluated to determine the efficacy of radiation therapy when used in a rescue therapy setting. All cats had relapsed or resistant lymphoma. Two fractions of radiation were delivered over 2 days for a total of 800cGy. Acute effects of radiation were not noted, except one cat that had a self-limiting loss of appetite. Response was noted in 10/11 cats. Median survival post-radiation therapy was 214 days and the overall median survival in this study was 355 days. This study suggests that abdominal irradiation for feline gastrointestinal lymphoma was well tolerated and may contribute to a positive clinical response.