Ensayos de bioacumulación de principios activos farmacológicos de Procambarus clarkii y Scrobicularia plana. Uso como bioindicadores en el Parque Natural de Doñana

En la presente Tesis Doctoral, se ha optimizado y validado un procedimiento analítico para la extracción y determinación simultánea de veintitrés principios activos farmacológicos, pertenecientes a muy diversas familias terapéuticas, en músculo abdominal y glándula digestiva de Procambarus clarkii. La extracción combina el uso simultáneo de una mezcla extractante acetonitrilo:agua y una digestión enzimática asistida por energía de microondas a baja potencia. La determinación de los extractos se realizó mediante cromatografía líquida de alta resolución acoplada a un detector de masas de triple cuadrupolo (HPLC-MS/MS). La metodología se tomó como referencia para optimizar y validar un procedimiento para la determinación de veintiséis principios activos farmacológicos mediante cromatografía líquida de alta resolución acoplada a un detector de masas de cuadrupolo-tiempo de vuelo (UPLC-MS-qTOF). Se llevó a cabo un estudio temporal de seis años sobre la presencia de analitos en especímenes de P.clarkii procedentes del Parque Nacional de Doñana empleando el procedimiento de HPLC-MS/MS triple cuadrupolo. Por último, se llevaron a cabo ensayos de exposición sobre los especímenes de P.clarkii y de S. plana sometidos a un cóctel de fármacos (flumequina, ciprofloxacina e ibuprofeno) a concentraciones de 10 y 100 µg L-1

Iconic Character of Bestiary Images in the Novels of F. Werfel and E. Canetti

The anthropomorphic depiction of animals bearing allegorical meanings is the well-represented and actively demanded bestiary of world literature. It reflects the mythological thinking of writers and is an integral part of the worldview basis on which literary works are based. Bestiary images in the artistic text acquire the status of universal representative symbols. This study discusses bestiary images in novelistic works of Austrian writers, Franz Werfel (1890-1945) and Elias Canetti (1905-1994). Using the semiotic approach, the researchers define a range of images and meanings that are related to these two writers as representatives of the era of historical upheavals and individual authorial purposes that reflect the basis of the worldview of each of them. A bestiary image in a literary text can function as an iconic sign, which, on the one hand, reflects the material object in its materiality, and on the other hand, contributes to the emerging of "new", constructed by analogy, aesthetic reality. The similarity to the referent, in this case, is included in the overall system of ontological values. An iconic sign, after Ch.S. Pierce, refers to a simple sign based on the similarity to a thing and participating in the creation of symptoms of a higher order – symbols. Bestiary images in a literary text acquire the status of universal representative characters. The functioning of animal images in the text, their nomination, combinatorics, communication with the elements, time periods, and behavioural patterns are the way of the study of the philosophical foundations of the author's world picture. Canetti’s bestiary is represented by metaphorical images of a monkey, a cat, a pig, and a tortoise, which are used as a tool for analysing various psychic and psychological states of the characters of the novel "Blinding" (Die Blendung, 1931-1932). Multiple forms of anthropopathy and zoomorphism are based on the writer's attitude towards the initial "equality" of man and animal. The study of zoopoetics (the term of J. Lacan) of Werfel’s novel "Barbara, or Piety" (Barbara oder die Frömigkeit, 1929) helps to reveal the axiological foundation upon which the writer constructs his novels. The functioning of images of animals, a horse, for example, is related to the semantics of sacrifice that is rethought and acquiring new meanings in a new historical context. Composite images connecting different characteristics and associated with various natural elements are important. It is apparent that the study of the works by Werfel and Canetti, given the iconic nature of bestiary images, seems relevant to detect common patterns of development of European literature and culture of the first third of the 20th Century

‘Unpacking’ technical attribution and challenges for ensuring stability in cyberspace

Submission to 2021–2025 UN Open-Ended Working Group (OEWG) on security of and in the use of information and communications technologies

Modern Austrian Novel: Endless Wanderings in the Labyrinths of Kafka's Castle

The purpose of the article is to discuss the perception of Kafka's The Castle (Das Schloss, 1922) in the novels The Peaches Killers (Die Pfirsichtöter, 1972) by Alfred Kolleritsch, Among the Bieresch (Bei den Bieresch, 1979) by Klaus Hoffer and Into the Castle (Ins Schloss, 2004) by Marianne Gruber. The reference to the writers and their works is no coincidence; preference is given to the artists whose creative manner reflects the most fashionable trends in Western European literary process - from avant-garde to postmodernism. The authors of the article deliberately arrange the analysed works in chronological order to follow the stages in the development of German postmodernism which originates from modernist literature. The universal Kafkaesque discourse suggests the existence of direct and inverse connections between the author and the reader, the extra-textual tradition and reality. The article focuses on the narrative strategies of Austrian avant-garde (Kolleritsch), analyses postmodern discourse (Hoffer, Gruber) in the Austrian literature of the second half of the 20th and early 21st centuries, reveals signs of typological similarity between the novels by Kafka, Kolleritsch, Hoffer, and Gruber, which seems productive for understanding the influence of modernist literature on the development of the postmodern paradigm in the German-language literary space. Austrian literature, to a greater extent, is fraught with the search for new forms of self-expression rather than with the artistic “overcoming the past” - the awareness of collective guilt.  It brought to the forefront the authors in whose works the age of change was reflected. Literary avant-garde has been replaced by authors who skillfully “play” with the previous culture and establish a dialogue with the present. The comparative methodology is to reveal the perception of "Kafkaesque discourse" in modern Austrian literature and to draw conclusions about the ways authors treat ontological questions

Macrophage scavenger receptors: Tumor support and tumor inhibition

Tumor-associated macrophages (TAMs) are a heterogeneous population of myeloid cells that constitute up to 50% of the cell mass of human tumors. TAMs interact with the components of the tumor microenvironment (TME) by using scavenger receptors (SRs), a large superfamily of multifunctional receptors that recognize, internalize and transport to the endosomal/lysosomal pathway apoptotic cells, cytokines, matrix molecules, lipid modified lipoproteins and other unwanted-self ligands. In our review, we summarized state-of-the art for the role of macrophage scavenger receptors in tumor development and their significance as cancer biomarkers. In this review we focused on functional activity of TAM-expressing SRs in animal models and in patients, and summarized the data for different human cancer types about the prognostic significance of TAM-expressed SRs. We discussed the role of SRs in the regulation of cancer cell biology, cell-cell and cell-matrix interaction in TME, immune status in TME, angiogenesis, and intratumoral metabolism. Targeting of tumor-promoting SRs can be a promising therapeutic approach in anti-cancer therapy. In our review we provide evidence for both tumor supporting and tumor inhibiting functions of scavenger receptors expressed on TAMs. We focused on the key differences in the prognostic and functional roles of SRs that are specific for cancer types. We highlighted perspectives for inhibition of tumor-promoting SRs in anti-cancer therapy

New angiogenic regulators produced by TAMs: perspective for targeting tumor angiogenesis

Angiogenesis is crucial to the supply of a growing tumor with nutrition and oxygen. Inhibition of angiogenesis is one of the main treatment strategies for colorectal, lung, breast, renal, and other solid cancers. However, currently applied drugs that target VEGF or receptor tyrosine kinases have limited efficiency, which raises a question concerning the mechanism of patient resistance to the already developed drugs. Tumor-associated macrophages (TAMs) were identified in the animal tumor models as a key inducer of the angiogenic switch. TAMs represent a potent source not only for VEGF, but also for a number of other pro-angiogenic factors. Our review provides information about the activity of secreted regulators of angiogenesis produced by TAMs. They include members of SEMA and S100A families, chitinase-like proteins, osteopontin, and SPARC. The COX-2, Tie2, and other factors that control the pro-angiogenic activity of TAMs are also discussed. We highlight how these recent findings explain the limitations in the efficiency of current anti-angiogenic therapy. Additionally, we describe genetic and posttranscriptional mechanisms that control the expression of factors regulating angiogenesis. Finally, we present prospects for the complex targeting of the pro-angiogenic activity of TAMs

PFKFB3 overexpression in monocytes of patients with colon but not rectal cancer programs pro-tumor macrophages and is indicative for higher risk of tumor relapse

IntroductionCirculating monocytes are main source for tumor-associated macrophages (TAMs) that control tumor growth, angiogenesis, metastasis and therapy resistance. We raised the questions how monocyte programming is affected by growing tumors localized in colon and rectal sections, and how treatment onsets affect monocyte programming in the circulation.MethodsPatients with rectal cancer and colon cancer were enrolled in the study. Peripheral blood monocytes were characterized by phenotypic analysis using flow cytometry, by transcriptomic analysis using RNA sequencing and by gene expression analysis using real-time RT-PCR. Phenotypic analysis was performed with IF/confocal microscopy. Spatial transcriptomic analysis was applied using GeoMX DSP-NGS.ResultsIn patients with rectal cancer, increased amount of CCR2+ monocytes was indicative for the absence of both lymphatic and hematogenous metastasis. In contrast, in patients with colon cancer CD163+ monocytes were indicative for LN metastasis. NGS analysis identified tumor-specific transcriptional programming of monocytes in all CRC patients compared to healthy individuals. The key transcriptional difference between monocytes of patients with colon and rectal cancer was increased expression of PFKFB3, activator of glycolysis that is currently considered as therapy target for major solid cancers. PFKFB3-expressing monocyte-derived macrophages massively infiltrated tumor in colon. Nanostring technology identified correlation of PFKFB3 with amount and tumor-promoting properties of TAMs in colon but not in rectal cancer. PFKFB3 was indicative for tumor relapse specifically in colon cancer.DiscussionOur findings provide essential argument towards CRC definition to cover two clinically distinct cancers – colon cancer and rectal cancer, that differentially interact with innate immunity

PFKFB3 overexpression in monocytes of patients with colon but not rectal cancer programs pro-tumor macrophages and is indicative for higher risk of tumor relapse

Introduction: Circulating monocytes are main source for tumor-associated macrophages (TAMs) that control tumor growth, angiogenesis, metastasis and therapy resistance. We raised the questions how monocyte programming is affected by growing tumors localized in colon and rectal sections, and how treatment onsets affect monocyte programming in the circulation. Methods: Patients with rectal cancer and colon cancer were enrolled in the study. Peripheral blood monocytes were characterized by phenotypic analysis using flow cytometry, by transcriptomic analysis using RNA sequencing and by gene expression analysis using real-time RT-PCR. Phenotypic analysis was performed with IF/confocal microscopy. Spatial transcriptomic analysis was applied using GeoMX DSP-NGS. Results: In patients with rectal cancer, increased amount of CCR2+ monocytes was indicative for the absence of both lymphatic and hematogenous metastasis. In contrast, in patients with colon cancer CD163+ monocytes were indicative for LN metastasis. NGS analysis identified tumor-specific transcriptional programming of monocytes in all CRC patients compared to healthy individuals. The key transcriptional difference between monocytes of patients with colon and rectal cancer was increased expression of PFKFB3, activator of glycolysis that is currently considered as therapy target for major solid cancers. PFKFB3-expressing monocyte-derived macrophages massively infiltrated tumor in colon. Nanostring technology identified correlation of PFKFB3 with amount and tumor-promoting properties of TAMs in colon but not in rectal cancer. PFKFB3 was indicative for tumor relapse specifically in colon cancer. Discussion: Our findings provide essential argument towards CRC definition to cover two clinically distinct cancers – colon cancer and rectal cancer, that differentially interact with innate immunity

Búsqueda de alimentos ricos en provitamina D (Ergosterol)

En este trabajo se presentan los resultados obtenidos para el contenido en provitamina D2 (ergosterol) en diferentes muestras de origen vegetal. Muestras de lechuga, dátiles, uvas, arroz y champiñones fueron sometidas a un tratamiento de extracción con hexano y el contenido en ergosterol medido mediante análisis por cromatografía líquida de alta resolución con detección ultravioleta de diodos en fila, obteniéndose los mayores contenidos en ergosterol en las muestras de champiñón y lechuga

Monitoring of pharmaceuticals in aquatic biota (Procambarus clarkii) of the Doñana National Park (Spain)

In this work the presence of different pharmaceuticals at Doñana National Park (Spain) and their main entry sources (input source or entry points) have been stated over the 2011–2016 years period. Twenty-three selected pharmaceuticals (corresponding to eight therapeutic families) were evaluated in crayfish and water samples from Doñana National Park (Spain) (six sampling points selected in order to cover different possible pollution sources into and surrounding the Park). The multiresidue determination was carried out using enzymatic-microwave assisted extraction prior to high performance liquid chromatography mass spectrometry detection. Sulphonamides (sulfadiazine, sulfamerazine, sulfamethazine, and sulfamethoxazole); trimethoprim, an antibiotic that is frequently co-administered with sulfamethoxazole; amphenicols (chloramphenicol, florfenicol and thiamphenicol); fluoroquinolones (ciprofloxacin, enrofloxacin, flumequine, danofloxacin, gatifloxacin, norfloxacin, marbofloxacin and grepafloxacin); penicillins (amoxicillin); tetracyclines (chlortetracycline and oxytetracycline); non-steroidal anti-inflammatory drugs (salicylic acid and ibuprofen); beta-blocker drugs (atenolol); and antiepileptics (carbamazepine) were analysed. Ciprofloxacin, ibuprofen, salicylic acid, flumequine, and carbamazepine were detected and/or quantified at some of the selected sampling points. A clear ecotoxicological risk to the ecosystem was demonstrated from the occurrence of ciprofloxacin in samples obtained after the punctual and massive presence of people inside the Park. Furthermore, flumequine and carbamazepine have been detected in Procambarus clarkii specimens in concentrations around 30 ng g−1 and 14 ng g−1, respectively, and their occurrence in the specimens could indicate the persistence of the discharge sources. The main source of pharmaceuticals into the Park might be the livestock farming activities, and the influence of urban wastewaters from surrounding villages does not seem to be very important.Dirección General de Investigación y Gestión del Plan Nacional de I+D+i (Ministerio de Economía y Competitividad) CTM2012-38720- C03-01, CTM2015-67902-C02-01-PFEDER/ Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación PGC2018-096608-B-C2