Trichome micromorphology in <Emphasis Type="Italic">Teucrium</Emphasis> (Lamiaceae) species growing in Croatia

Micromorphological investigation of the types, dimensions and distribution of characteristic trichomes in leaves and stems in Teucrium L. species (T. arduini L., T. chamaedrys L., T. flavum L., T. montanum L., T. polium L., and T. scordium L. subsp. scordioides Schreb.) distributed in Croatia was carried out as part of the taxonomical study of the genus Teucrium. Secretory types of hairs, peltate and capitate hairs were observed on the epidermis of stems and leaves of all investigated species. Non-secretory, acicular hairs were almost completely lacking on stems of T. scordium subsp. scordioides. Flagelliform hairs were not found in T. flavum and T. polium. Cladose hairs were present only in T. polium. The largest micromorphological variability was established between wild and cultivated samples of T. arduini and T. scordium subsp. scordioides, while cultivated and wild specimens of T. polium were almost identical. Differences were primarily observed in trichome dimensions and much less in micromorphological features.     

A new species of <Emphasis Type="Italic">Isospora</Emphasis> Schneider, 1881 (Apicomplexa: Eimeriidae) in Ruppell’s agama <Emphasis Type="Italic">Agama rueppelli</Emphasis> (Vaillant) (Sauria: Agamidae) from East Africa,with a review of this genus in agamid lizards

Coprological examinations of eight Ruppell’s agamas Agama rueppelli (Vaillant) revealed the presence of a coccidium of the genus Isospora Schneider, 1881 that represents a previously undescribed species. Oöcysts of Isospora farahi n. sp. are spherical or subspherical, 29.1 (26–31) × 28.8 (26–31) μm, with a shape-index of 1.01 (1–1.07). An oöcyst residuum, polar granules and micropyle are absent. The oöcyst wall is bilayered, brownish and smooth, c. 1.5–2 μm thick. The sporocysts are oval, 16.6 (15–18) × 11.4 (11–12) μm, with a shape-index of 1.46 (1.25–1.64) and both Stieda and substieda bodies. A sporocyst residuum is present as medium-sized granules scattered irregularly among the sporozoites. The sporozoites are vermiform, with a large posterior spherical refractile body. Endogenous development is intranuclear in the epithelial cells of the small intestine. Sporulation is unknown, as oöcysts were recovered from the faeces.     

Mapping field-scale spatial patterns of size and activity of the denitrifier community

There is ample evidence that microbial processes can exhibit large variations in activity on a field scale. However, very little is known about the spatial distribution of the microbial communities mediating these processes. Here we used geostatistical modelling to explore spatial patterns of size and activity of the denitrifying community, a functional guild involved in N-cycling, in a grassland field subjected to different cattle grazing regimes. We observed a non-random distribution pattern of the size of the denitrifier community estimated by quantification of the denitrification genes copy numbers with a macro-scale spatial dependence (6–16 m) and mapped the distribution of this functional guild in the field. The spatial patterns of soil properties, which were strongly affected by presence of cattle, imposed significant control on potential denitrification activity, potential N₂O production and relative abundance of some denitrification genes but not on the size of the denitrifier community. Absolute abundance of most denitrification genes was not correlated with the distribution patterns of potential denitrification activity or potential N₂O production. However, the relative abundance of bacteria possessing the nosZ gene encoding the N₂O reductase in the total bacterial community was a strong predictor of the N₂O/(N₂ + N₂O) ratio, which provides evidence for a relationship between bacterial community composition based on the relative abundance of denitrifiers in the total bacterial community and ecosystem processes. More generally, the presented geostatistical approach allows integrated mapping of microbial communities, and hence can facilitate our understanding of relationships between the ecology of microbial communities and microbial processes along environmental gradients.     

Influenza A Virus Induces an Immediate Cytotoxic Activity in All Major Subsets of Peripheral Blood Mononuclear Cells

Background

A replication defective influenza A vaccine virus (delNS1 virus) was developed. Its attenuation is due to potent stimulation of the innate immune system by the virus. Since the innate immune system can also target cancer cells, we reasoned that delNS1 virus induced immune-stimulation should also lead to the induction of innate cytotoxic effects towards cancer cells.

Methodology/Principal Findings

Peripheral blood mononuclear cells (PBMCs), isolated CD56+, CD3+, CD14+ and CD19+ subsets and different combinations of the above subsets were stimulated by delNS1, wild type (wt) virus or heat inactivated virus and co-cultured with tumor cell lines in the presence or absence of antibodies against the interferon system. Stimulation of PBMCs by the delNS1 virus effectively induced cytotoxicity against different cancer cell lines. Surprisingly, virus induced cytotoxicity was exerted by all major subtypes of PBMCs including CD56+, CD3+, CD14+ and CD19+ cells. Virus induced cytotoxicity in CD3+, CD14+ and CD19+ cells was dependent on virus replication, whereas virus induced cytotoxicity in CD56+ cells was only dependent on the binding of the virus. Virus induced cytotoxicity of isolated cell cultures of CD14+, CD19+ or CD56+ cells could be partially blocked by antibodies against type I and type II (IFN) interferon. In contrast, virus induced cytotoxicity in the complete PBMC preparation could not be inhibited by blocking type I or type II IFN, indicating a redundant system of activation in whole blood.

Conclusions/Significance

Our data suggest that apart from their well known specialized functions all main subsets of peripheral blood cells also initially exert a cytotoxic effect upon virus stimulation. This closely links the innate immune system to the adaptive immune response and renders delNS1 virus a potential therapeutic tool for viro-immunotherapy of cancer.     

Interstitial Cajal-like cells of human Fallopian tube express estrogen and progesterone receptors

Cells of the female reproductive tract are subject to hormonal control via sex steroid genomic receptors expressed at nuclear level. We previously showed that interstitial Cajal-like cells (ICLC) of human myometrium expressed estrogen and progesterone receptors (ER/PR). Our aim, based on these results, was to see if ER and/or PR could be found also in tubal ICLC. Indeed, we present here immunohistochemical evidence that ICLC of human Fallopian tube (isthmic region) have such receptors. Stromal ICLC, as well as ICLC among smooth muscle layers, were identified in tissue sections by their morphological features (e.g. several very long, moniliform, prolongations of cell body) as well as by c-kit positivity, vital staining with methylene blue or silver impregnation. Additional evidence was provided by sequential staining for c-kit and for PR on the same cell, by ‘sandwich method’. In vitro, the 4th passage cell cultures from Fallopian tube muscularis exhibiting ICLC morphology showed the presence of ER-alpha and/or PR-A by immunofluorescence. In conclusion, our data suggest that ICLC could function as steroid sensors, and might be implicated in Fallopian tube motility (via gap junctions or juxta- and/or paracrine mechanisms).     

Phospholemman phosphorylation alters its fluorescence resonance energy transfer with the Na/K-ATPase pump

Phospholemman (PLM) or FXYD1 is a major cardiac myocyte phosphorylation target upon adrenergic stimulation. Prior immunoprecipitation and functional studies suggest that phospholemman associates with the Na/K-pump (NKA) and mediates adrenergic Na/K-pump regulation. Here, we tested whether the NKA-PLM interaction is close enough to allow fluorescence resonance energy transfer (FRET) between cyan and yellow fluorescent (CFP/YFP) fusion proteins of Na/K pump and phospholemman and whether phospholemman phosphorylation alters such FRET. Co-expressed NKA-CFP and PLM-YFP in HEK293 cells co-localized in the plasma membrane and exhibited robust FRET. Selective acceptor photobleach increased donor fluorescence (F(CFP)) by 21.5 +/- 4.1% (n = 13), an effect nearly abolished when co-expressing excess phospholemman lacking YFP. Activation of protein kinase C or A progressively and reversibly decreased FRET assessed by either the fluorescence ratio (F(YFP)/F(CFP)) or the enhancement of donor fluorescence after acceptor bleach. After protein kinase C activation, forskolin did not further reduce FRET, but after forskolin pretreatment, protein kinase C could still reduce FRET. This agreed with phospholemman phosphorylation measurements: by protein kinase C at both Ser-63 and Ser-68, but by protein kinase A only at Ser-68. Expression of PLM-YFP and PLM-CFP resulted in even stronger FRET than for NKA-PLM (F(CFP) increased by 37 +/- 1% upon YFP photobleach), and this FRET was enhanced by phospholemman phosphorylation, consistent with phospholemman multimerization. Co-expressed PLM-CFP and Na/Ca exchange-YFP were highly membrane co-localized, but FRET was undetectable. We conclude that phospholemman and Na/K-pump are in very close proximity (FRET occurs) and that phospholemman phosphorylation alters the interaction of Na/K-pump and phospholemman.     

Health risks associated with wild animal translocation: a case of the European bison and an alien parasite

Introducing an animal into a new location could be hazardous in the form of disease transmission, especially with respect to infections that are often overlooked. Such introduced infectious agents, including parasitic ones, then pose potential danger to the native animal population. Within the conservation program, the European bison was introduced into many European countries. However, this largest European herbivore was recognized as a new host for an invasive parasitic nematode, Ashworthius sidemi, in Poland in 1998. Since then, the prevalence of this non-native parasite in Poland has increased not only in bison but also in other wild ruminants. In 2011 five European bison individuals were transported from Poland to the Czech Republic. In the current study, we examined the gastrointestinal tracts of two European bison and two red deer culled in the ?idlov game reserve. A. sidemi was identified in all investigated animals using both morphological and molecular methods; infection intensity was higher in bison than in deer. Our findings represent the first record of this invasive parasite in European bison at the Czech territory. The results of this study indicate changes in epidemiological patterns of Ashworthius infections in the climatic condition of Central Europe as well as the need to verify the reliability of ashworthiosis intravital diagnostics. One can expect A. sidemi to spread gradually in the Czech Republic and colonize other native ruminant hosts.