The use of subjunctive/indicative with operators of probability modality in Latin American Spanish (central, Mexican and Caribian areas).

The purpose of this bachelor thesis is to verify the use of subjunctive/indicative after operator quizás in all of its possible variations, and also taking into account regional usage differences in Latin American Spanish, focusing on the Central area (Costa Rica, Nicaragua) and the Caribbean area (Cuba, Puerto Rico, Panama and Dominican Republic). The introduction deals with the theoretical aspect of this grammar phenomenon and the rest of the thesis concentrates on statistical research which has been developed with the help of the Royal Spanish Academy's corpus CREA

Alteration: Diminutives and Augmentatives in Contemporary Italian

This thesis deals with alteration: diminutives and augmentatives in contemporary Italian, and their pragmatic functions. The aim of this thesis is to create a list of diminutives and augmentatives used in Italian fiction. The analysis of expressions is based on the texts contained in InterCorp. The starting point is a theoretical introduction to the word formation with special attention to alterative suffixes in Italian and Czech language, with a partial comparison to Spanish. The thesis also concentrates on hypocorisms, phonological and suffixal restrictions, and lexicalization of alteratives. An integral part is the chapter dedicated to pragmatics, speech acts theory and pragmatic use of alterative suffixes. The results of this thesis represent concrete illustrations for various communication situations documented from corpus

Epistemic Modality in Specialized Spanish and Czech Texts: A Comparative Analysis

The thesis deals with the epistemic modality, its types and modes of expression in specialized Spanish and Czech texts. The aim of this thesis is to perform a comparative analysis which is based on a statistical analysis of linguistic texts. The introduction primarily concentrates on the different typologies of Czech and foreign authors, the classification of modal attitudes, and the lexical and grammatical means of expressing modality. The result of this thesis is the comparison of the frequency that can be observed between the aforementioned languages

Are Histidine Kinases of Arbuscular Mycorrhizal Fungi Involved in the Response to Ethylene and Cytokinins?

International audienceSignals are exchanged at all stages of the arbuscular mycorrhizal (AM) symbiosis between fungi and their host plants. Root-exuded strigolactones are well-known early symbiotic cues, but the role of other phytohormones as interkingdom signals has seldom been investigated. Here we focus on ethylene and cytokinins, for which candidate receptors have been identified in the genome of the AM fungus Rhizophagus irregularis. Ethylene is known from the literature to affect asymbiotic development of AM fungi, and in the present study, we found that three cytokinin forms could stimulate spore germination in R. irregularis. Heterologous complementation of a Saccharomyces cerevisiae mutant strain with the candidate ethylene receptor RiHHK6 suggested that this protein can sense and transduce an ethylene signal. Accordingly, its N-terminal domain expressed in Pichia pastoris displayed saturable binding to radiolabeled ethylene. Thus, RiHHK6 displays the expected characteristics of an ethylene receptor. In contrast, the candidate cytokinin receptor RiHHK7 did not complement the S. cerevisiae mutant strain or Medicago truncatula cytokinin receptor mutants and seemed unable to bind cytokinins, suggesting that another receptor is involved in the perception of these phytohormones. Taken together, our results support the hypothesis that AM fungi respond to a range of phytohormones and that these compounds bear multiple functions in the rhizosphere beyond their known roles as internal plant developmental regulators. Our analysis of two phytohormone receptor candidates also sheds new light on the possible perception mechanisms in AM fungi

Pepsin digest of wheat gliadin fraction increases production of IL-1β via TLR4/MyD88/TRIF/MAPK/NF-κB signaling pathway and an NLRP3 inflammasome activation.

Celiac disease (CD) is a gluten-responsive, chronic inflammatory enteropathy. IL-1 cytokine family members IL-1β and IL-18 have been associated with the inflammatory conditions in CD patients. However, the mechanisms of IL-1 molecule activation in CD have not yet been elucidated. We show in this study that peripheral blood mononuclear cells (PBMC) and monocytes from celiac patients responded to pepsin digest of wheat gliadin fraction (PDWGF) by a robust secretion of IL-1β and IL-1α and a slightly elevated production of IL-18. The analysis of the upstream mechanisms underlying PDWGF-induced IL-1β production in celiac PBMC show that PDWGF-induced de novo pro-IL-1β synthesis, followed by a caspase-1 dependent processing and the secretion of mature IL-1β. This was promoted by K+ efflux and oxidative stress, and was independent of P2X7 receptor signaling. The PDWGF-induced IL-1β release was dependent on Nod-like receptor family containing pyrin domain 3 (NLRP3) and apoptosis-associated speck like protein (ASC) as shown by stimulation of bone marrow derived dendritic cells (BMDC) from NLRP3(-/-) and ASC(-/-) knockout mice. Moreover, treatment of human PBMC as well as MyD88(-/-) and Toll-interleukin-1 receptor domain-containing adaptor-inducing interferon-β (TRIF)(-/-) BMDC illustrated that prior to the activation of caspase-1, the PDWGF-triggered signal constitutes the activation of the MyD88/TRIF/MAPK/NF-κB pathway. Moreover, our results indicate that the combined action of TLR2 and TLR4 may be required for optimal induction of IL-1β in response to PDWGF. Thus, innate immune pathways, such as TLR2/4/MyD88/TRIF/MAPK/NF-κB and an NLRP3 inflammasome activation are involved in wheat proteins signaling and may play an important role in the pathogenesis of CD

PDWGF stimulates BMDC to IL-1β production through NLRP3 and ASC.

<p>(<b>A</b>) BMDC from WT, NLRP3−/− and ASC−/− mice were exposed to PDWGF (100 µg/ml) alone for 24 h; or first PDWGF was added for 21.5 h, the subsequently ATP (2 mM) was added for additional 2.5 h. IL-1β was measured in culture supernatants. (<b>B</b>) Flow-cytometric evaluation of PDWGF-induced maturation assessed by CD40, CD80, and CD86 expression on BMDC from WT and NLRP3−/− mice. WT and NLRP3−/− BMDC were cultured with 100 µg/ml of PDWGF (green), as well as 0.1 µg/ml of LPS (red) or 100 µg/ml of OVA (grey-filled) as positive and negative controls, respectively. Isotype controls are represented in black overlays. (<b>C</b>) Cells were preincubated with caspase-1 inhibitor Z-YVAD-fmk for 30 min, and then exposed to PDWGF in combination with ATP. Production of IL-1β was measured in culture supernatants. Results are expressed as mean ± SD from 4 independent experiments. The levels of significance for KO BMDC vs. WT BMDC are indicated as follows: *P<0.05, **P<0.01, and ***P<0.001.</p

TLR signaling is required for pro-IL-1β synthesis in response to PDWGF.

<p>WT BMDC and MyD88−/−, TRIF−/−, and IL-1R−/− KO BMDC were treated with PDWGF alone or in combination with ATP and (<b>A</b>) IL-1β production was evaluated after 24 h. (<b>B</b>) Cell lysates were evaluated for <i>de novo</i> synthesis of pro-IL-1β (<b>C</b>) WT BMDC and TLR2−/−, TLR4−/−, and TLR2/4−/− KO BMDC were treated with PDWGF alone or in combination with ATP, and IL-1β production was evaluated after 24 h. (<b>D</b>) Cell lysates were evaluated for <i>de novo</i> synthesis of pro-IL-1β. Data in (A) and (C) are expressed as mean ± SD from 5 independent experiments. *P<0.05, **P<0.01 vs. WT BMDC. Blots in (B) and (D) are representative from 3 independent experiments. β-actin was used as a loading control. (<b>E</b>) Celiac PBMC were treated with PDWGF alone or in combination with anti-TLR4 or anti-TLR2 Ab. IL-1β secretion was evaluated after 24 h. LPS was used as a positive control. Mean ± SD, 8 independent experiments. **P<0.01, ***P<0.001 vs. cells without anti-TLR Ab.</p

Genotype frequencies for <i>NALP1</i> (rs12150220) and <i>NALP3</i> (rs10754558).

<p>OR- odds ratio, CI- confidence interval.</p

PDWGF digest induces IL-1β, IL-18, and IL-1α release in monocytes and PBMC from CD patients.

<p>IL-1β (A), IL-18 (B) and IL-1α (C) levels were quantified in cell supernatants by ELISA. Data are given as mean ± SD from 39 patients and 15 healthy donors (HD). *P<0.05, **P<0.01 (CD vs. HD). (<b>D</b>). PDWGF-induced activation of PBMC is not due to LPS contamination. Results are shown as the percentage of the cytokine production from 4 CD patients. The data were normalized to the result from untreated cells which was set as 100%. Mean ± SD, n = 4 independent experiments, ***P<0.001 compared to untreated cells.</p

PDWGF-induced IL-1β production from celiac patient PBMC is modulated by K+ efflux, but is independent of the P2X7 receptor; as shown by (A) ELISA, mean ± SD, n = 10, ***P<0.001 vs. PDWGF-treated cells; and by (B) Western blot.

<p>Representative blots from 5 independent experiments are shown. (<b>C</b>) Inhibition of ROS modulate PDWGF-induced IL-1β secretion, mean ± SD, n = 10; as well as (D) pro-IL-1β production from PBMC of CD patients. Representative blots from 3 independent experiments are shown. β-actin was used as a loading control. ***P<0.001 vs. PDWGF-treated cells.</p