<p dir="ltr"><b>Background</b></p><p dir="ltr">Heart failure (HF) is a chronic clinical syndrome characterized by symptoms including breathlessness, fatigue, swelling of the ankles, and signs such as edema pulmonary crackles etc. During HF, pathogenic mechanisms including hemodynamic overload, ventricular remodeling, aberrant calcium handling, excessive neurohormonal stimulation contribute to the worsening and progression of the condition. Ventricular arrhythmias are the common cause of sudden cardiac death (SCD) in HF patients.</p><p dir="ltr">Hyperactivation of the sympathetic nervous system (SNS), a characteristic of HF, causes an increase in circulating catecholamines which becomes detrimental to-adrenergic receptors (-AR) leading to signaling dysfunction, and decrease in contractility and the ionotropic reserve. Expression of calcium/calmodulin-dependent protein kinase II (CaMKII), a downstream effector of-AR and a key regulator of calcium homeostasis, has been shown to be enhanced in HF. CaMKII-mediated mechanisms have been demonstrated to contribute to cardiac remodeling, arrhythmias by pathological regulation of ion channels, and contractile dysfunction.</p><p dir="ltr">The human ether-a-go-go related gene (hERG) encodes the pore-forming subunit of the voltage-gated potassium channel that conduct the rapid component of the delayed rectifier potassium current, <i>I</i>_Kr. The gating kinetics of <i>I</i>_Krmakes it a crucial determinant of the duration of the plateau phase of atrial and ventricular action potential (AP). Reduced <i>I</i>_Kr density due to loss-of-function mutations or pharmacological blockage of hERG channels precipitate arrhythmias. Downregulation of <i>I</i>_Kr density and protein have been reported in HF. Recent studies suggest that microRNAs (miRNAs) are involved in pathological downregulation of hERG.</p><p dir="ltr">miRNA are small non-coding RNAs of approximately 22 nucleotides in length that function as gene expression regulatory elements by repression translation. Aberrant miRNA expression has associated with cancer, cardiovascular, autoimmune, and inflammatory disorders.</p><p dir="ltr"><b>Objective</b></p><p dir="ltr">The overarching objective of this study is to investigate the mechanisms of CaMKII-mediated regulation of hERG function, including assessment of an interplay with miR-362-3p during sustained β-AR stimulation. In Specific Aim 1, the effect of CaMKII activation through sustained β-AR stimulation on hERG function and miR-362-3p expression will be assessed. The mechanism of miR-362-3p upregulation will be evaluated in Specific Aim 2, and in Specific Aim 3, the interactome of miR-362-3p and binding sites will be characterized and predicted, respectively.</p><p dir="ltr"><b>Methods</b></p><p dir="ltr">Whole-cell, voltage clamp electrophysiology experiments were performed in HEK 293 cells stably expressing hERG (hERG-HEK) and both hERG and wild-type CaMKIIδ<br>(hERG/CaMKII-HEK) following treatment with isoproterenol for 48 hours, and after transfection with miR-362-3p. The effect of CaMKII activation on miR-362-3p was assessed using real-time quantitative polymerase chain reaction (RT-qPCR). Total RNA was isolated 48 hours after isoproterenol treatment and the TaqMan assay was used to reverse transcribe and analyze miR-362-3p expression. Cells were transfected with cJun siRNA and precursor miR-362-3p to assess the role of cJun miR-362-3p upregulation during sustained β-AR stimulation with isoproterenol. The interactome of miR-362-3p was assessed in both cell lines using enhanced crosslinking immunoprecipitation (eCLIP) assay. miR-362-3p binding sites were predicted using RNAStructure Duplexfold after identification of miR-362-3p chimeric molecules from eCLIP experiment. Interaction analysis was performed using GeneMania in Cytoscape to identify genes that were potentially downregulated by miR-362-3p and been reported to interact with hERG.</p><p dir="ltr"><b>Results</b></p><p dir="ltr">In Specific Aim 1, the effect of sustained β-AR stimulation on hERG currents and endogenous miR-362-3p was assessed in hERG-HEK and hERG/CaMKII-HEK cells. Using whole-cell voltage clamp electrophysiology, we demonstrated that 48 hours treatment with 100 nM isoproterenol reduced hERG currents in hERG/CaMKII-HEK cells (p = 0.032) but had no effect on the voltage dependence of activation (p = 0.61) relative to control vehicle. Isoproterenol treatment for 48 hours, however, had no effect on hERG currents (p = 0.58) and the voltage dependence of activation (p = 0.99) in hERG-HEK cells. The effect of sustained isoproterenol treatment on miR-362-3p was also assessed using RT-qPCR. In hERG/CaMKII cells, 48 hours isoproterenol treatment increased miR-362-3p expression (2.3 folds; p = 0.038) relative to control vehicle. hERG/CaMKII-HEK cells were also treated with 500 nM KN-93 or its inactive analogue, KN-92, in an attempt to reverse CaMKII effect on miR-362-3p expression. Treatment with KN-93 decreased miR-362-3p expression (0.5-fold; p = 0.002) relative KN-92 treatment. Isoproterenol treatment had no effect on miR-362-3p expression in hERG-HEK cells (p = 0.38).</p><p dir="ltr">The regulatory mechanism of miR-362-3p expression was evaluated in Specific Aim 2. The role of an activator protein-1 (AP-1)-like sequence located at 98 base pairs upstream of miR-362-3p transcription start site was probed using siRNA inhibition of cJun, a central protein of the AP-1 complex, and deletion of the site sequence. The effect of exogenous miR-362-3p on hERG currents were first assessed. Precursor miR-362-3p decreased hERG currents (p = 0.003) compared to control plasmid. The effect of CaMKII overexpression was also assessed on exogenous miR-363-3p expression. Isoproterenol treatment in hERG/CaMKII-HEK cells transfected with precursor miR-362-3p increased mature miR-362-3p expression (0.029) compared to control vehicle treatment. Inhibition of cJun inhibition with cJun-specific siRNA decreased mature miR-362-3p expression (0.5-fold; p = 0.027) compared to scramble siRNA in hERG-HEK cells. In hERG-HEK cells transfected with mutated precursor miR-362-3p (AP-1-like site deleted), cJun inhibition with siRNA had no effect on miR-362-3p expression (p = 0.40).</p><p dir="ltr">The focus of Specific Aim 3 was to characterize the interactome of miR-362-3p as well as predict the miRNA response element (MRE) of its target mRNAs using enhanced crosslinking immunoprecipitation. A network analysis was also performed to identify miR-362-3p targets that have been reported to interact with hERG. Approximately 23% of miR-362-3p mRNA targets from the eCLIP assay have also been catalogued in miRNA database, TargetScanHuman, as miR-362-3p targets. miR-362-3p chimeric molecules with 853 unique targets, of which 75 were identified to interact with hERG through the network analysis. Four unique chimeric molecules between miR-362-3p and hERG mRNA were identified, but the interactions were non-canonical (located in the coding sequence of hERG and outside the seed region of miR-362-3p). Thirty five of the 75 miR-362-3p targets that were identified to interact had a chimeric read ≥ 3, a cutoff number indicating non-random chimeric formation. Using RNAStructure DuplexFold, miR-362-3p was predicted to form canonical binding with 12 of 35 mRNA targets. HSPA4, a heat shock protein involved in the maturation and trafficking of hERG, was identified in a canonical interaction (8-mer) with miR-362-3p.</p><p dir="ltr"><b>Conclusion</b>:</p><p dir="ltr">Sustained β-AR stimulation increases miR-362-3p expression and decreases hERG currents in CaMKII overexpressing cells. cJun mediates miR-362-3p upregulation by interacting with an AP-1-like sequence upstream of miR-362-3p transcription start site. Pathological regulation of <i>I</i>_Kr by CaMKII mediated by miR-362-3p during sustained-AR may contribute to increased risk of arrhythmias in states of increase catecholaminergic activity, such as HF.</p>

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No description available.

Perceptual features are qualitative features used to describe music properties in relation to human perception instead of typical musical theory concepts such as pitches and chords. This report describes a music discovery platform which uses three different methods of music playlist generation to investigate if and how perceptual features work when used for music discovery. One method abstracts away the complexity of perceptual features and the other two lets users use them directly. Two user testing sessions were performed to evaluate the browser and compare the different methods. Test participants found the playlist generation to work well in general, and especially found the method which uses emotions as an interface to be intuitive, enjoyable and something they would use to find new music. The other two methods which let users directly interact with perceptual features were less popular, especially among users without musical education. Overall, using perceptual features for music discovery was successful, although methods should be chosen with the intended audience in mind. / Perceptuella särdrag är kvalitativt framtagna särdrag som beskriver musik med fokus på mänsklig perception snarare än musikteoribegrepp som tonhöjd och ackord. Den här rapporten beskriver en musikhemsida som använder tre olika metoder för att generera spellistor med avsikt att undersöka om och hur perceptuella särdrag fungerar för att hitta ny musik. En metod abstraherar bort perceptuella särdragens komplexitet och de andra två metoderna låter testare använda dem utan abstraktion. Två användbarhetstest utfördes för att utvärdera musikhemsidan och jämföra de olika metoderna. Testanvändare tyckte överlag att genereringen av spellistor fungerade bra och att speciellt metoden som använde känslor som gränssnitt var intuitiv, rolig att använda och en metod de skulle använda för att hitta ny musik. De andra två metoderna som tillät användare att direkt använda perceptuella särdrag var mindre populära, speciellt bland användare utan musikutbildning. Överlag var användandet av perceptuella särdrag för att hitta musik en framgång, dock bör metoderna väljas utifrån användarnas kunskap.

[ES] La presente tesis doctoral, titulada "papel del miR-99a-5p en cáncer de mama: transformando hallazgos moleculares en herramientas clínicas" está centrada en el estudio del microRNA miR-99a-5p en cáncer de mama y en la determinación de su potencial como diana terapéutica y biomarcador para el diagnóstico de dicha enfermedad. En el primer capítulo, se lleva a cabo una revisión general del cáncer de mama y los microRNAs. A continuación, en el segundo capítulo se presentan los objetivos. En el tercer capítulo se compararon los perfiles de expresión de microRNAs de una línea celular de cáncer de mama resistente a doxorrubicina frente a la línea celular parental, dando lugar a la identificación del microRNA miR-99a-5p como el más desregulado entre ambas. A continuación, se confirmó que el miR-99a-5p aumenta la sensibilidad a doxorrubicina mediante la inhibición directa de la expresión de COX-2, que conlleva la inhibición de la expresión de la proteína ABCG2, ampliamente descrita por su papel en resistencia a fármacos. En base a los resultados obtenidos, se diseñaron nanodispositivos basados en nanopartículas mesoporosas de sílice para la liberación de miR-99a-5p y doxorrubicina en los tumores. Se confirmó la eficacia de las nanopartículas para reducir el crecimiento tumoral así como los efectos adversos asociados a la doxorrubicina libre en un modelo ortotópico murino de cáncer de mama. En el cuarto capítulo, se evaluó el potencial del miR-99a-5p como biomarcador para el diagnóstico del cáncer de mama. Se determinó la expresión del microRNA en tumores primarios de cáncer de mama y en tejidos sanos. Los resultados mostraron que el miR-99a-5p se encuentra infraexpresado en los tejidos cancerosos. A continuación, con el objeto de determinar el potencial del miR-99a-5p como biomarcador mínimamente invasivo, se determinaron los niveles de expresión en plasma de pacientes de cáncer de mama y de donantes sanos. Se encontró que el miR-99a-5p se encuentra a mayor concentración en el plasma de pacientes con cáncer de mama. Estos resultados se validaron en una cohorte independiente. Mediante el análisis en base a curvas ROC se confirmó que el miR-99a-5p es útil como biomarcador no invasivo para el diagnóstico del cáncer de mama incluso en estadíos tempranos de la enfermedad. El quinto capítulo se centra el diseño y validación de un biosensor basado en soportes de alúmina mesoporosa para la detección del miR-99a-5p en plasma. Los poros de una placa de alúmina mesoporosa se cargaron con el fluoróforo rodamina B, y se utilizaron oligonucleótidos complementarios a la secuencia del miR-99a-5p como puertas moleculares capaces de retener los fluoróforos en el interior de los poros. En presencia del miR-99a-5p, se produce liberación de rodamina B, que es posteriormente detectada ópticamente. Se confirmó la especificidad del biosensor, así como su alta sensibilidad. Además, utilizando muestras de plasma de pacientes de cáncer de mama y controles sanos, se confirmó la eficiencia del dispositivo para detectar cáncer de mama incluso en estadíos tempranos. Por último, en los capítulos cinco y seis se presentan la discusión general y las conclusiones principales extraídas. En conclusión, esta tesis doctoral demuestra que el miR-99a-5p tiene potencial como diana terapéutica y biomarcador para el cáncer de mama, y podría ser útil para mejorar el pronóstico de los pacientes con dicha enfermedad. Se ha elucidado que uno de los mecanismos por los que el miR-99a-5p está involucrado en sensibilidad a doxorrubicina es mediante la regulación del eje COX-2/ABCG2. Además, las nanopartículas diseñadas para la administración combinada de miR-99a-5p y doxorrubicina se presentan como una herramienta con gran potencial para el tratamiento oncológico. Por último, aprovechando el potencial del miR-99a-5p como biomarcador diagnóstico, se ha desarrollado un sistema de detección que podría ser una herramienta útil para mejorar la detección temprana del cáncer de mama. / [CA] La present tesis doctoral, titulada "paper del miR-99a-5p en càncer de mama: transformant descobriments moleculars en eines clíniques" està centrada en l'estudi del microRNA miR-99a-5p en càncer de mama i en la determinació del seu potencial com a diana terapèutica i biomarcador per al diagnòstic d'aquesta malaltia. En el primer capítol, es du a terme una revisió general del càncer de mama i els microRNAs. A continuació, en el segon capítol es presenten els objectius. En el tercer capítol es van comparar els perfils d'expressió de microRNAs d'una línia cel·lular de càncer de mama resistent a doxorrubicina amb la seua línia parental, donant lloc a la identificació del microRNA miR-99a-5p com el més desregulat entre ambdues. A continuació, es va confirmar que el miR-99a-5p augmenta la sensibilitat al fàrmac mijançant la inhibició directa de l'expressió de COX-2 de forma directa, que comporta la inhibició de l'expressió de la proteïna ABCG2 que està àmpliament descrita pel seu paper en resistència a fàrmacs. Basant-se en els resultats obtinguts, es van dissenyar nanodispositius basats en nanopartícules mesoporoses de sílice per a l'alliberament d'una combinació de miR-99a-5p i doxorrubicina en els tumors. L'eficacia dels sistemes per a reduir el creixement tumoral així com els efectes adversos associats a la doxorrubicina lliure es va confirmar en un model ortotòpic murí de càncer de mama. En el quart capítol, es va avaluar el potencial del miR-99a-5p com a biomarcador per al diagnòstic del càncer de mama. Es va determinar l'expressió del microRNA en tumors primaris de càncer de mama i en teixits sans. Els resultats van mostrar que el miR-99a-5p es troba infraexpressat en els teixits cancerosos. A continuació, amb l'objectiu de determinar el potencial del miR-99a-5p com a biomarcador mínimament invasiu, es van determinar els nivells d'expressió en plasma de pacients de càncer de mama i de donants sans. Es va trobar que el miR-99a-5p es troba a major concentració en el plasma de pacients amb càncer de mama. Aquestos resultats es van validar en una cohort independent. Per mitjà de l'anàlisi basant-se en corbes ROC es va confirmar que el miR-99a-5p és útil com a biomarcador no invasiu per al diagnòstic del càncer de mama inclús en estadis primerencs de la malaltia. El quint capítol es centra en el disseny i validació d'un biosensor basat en suports d'alúmina nanoporosa per la detecció del miR-99a-5p en plasma. Els porus d'una placa d'alúmina es van carregar amb l'indicador fluorescent rodamina B, i es van utilitzar oligonucleòtids complementaris a la seqüència del miR-99a-5p com a portes moleculars amb la capacitat de retindre els fluoròfors a l'interior dels porus. En presència del miR-99a-5p, es produeix l'alliberament de rodamina B, que serà posteriorment detectada òpticament. Es va confirmar l'especificitat del biosensor, així com la seua alta sensibilitat. A més, utilitzant mostres de plasma de pacients de càncer de mama i controls sans, es va confirmar l'eficiència del dispositiu per a detectar càncer de mama inclús en estadis primerencs. Finalment, en els capítols sis i set es presenten la discussió general i les conclusions principals. En conclusió, aquesta tesi doctoral demostra que el miR-99a-5p té potencial com a diana terapèutica i biomarcador per al càncer de mama, i podria ser útil per a millorar el pronòstic dels pacients amb dita malaltia. S'ha elucidat que un dels mecanismes pels quals el miR-99a-5p està involucrat en sensibilitat a doxorrubicina és per mitjà de la regulació de l'eix COX-2/ABCG2. A més, les nanopartícules dissenyades per a l'administració combinada de miR-99a-5p i doxorrubicina es presenta com una ferramenta amb gran potencial per al tractament oncològic. Finalment, aprofitant el potencial del miR-99a-5p com a biomarcador diagnòstic, s'ha desenvolupat un sistema de detecció que podria ser una ferramenta útil per a millorar la detecció primerenca del càncer de mama. / [EN] This PhD thesis, entitled "Role of miR-99a-5p in breast cancer: translating molecular findings into clinical tools" is focused on the study of the microRNA miR-99a-5p in breast cancer and in determining its potential as a therapeutic target and biomarker for the diagnosis of this disease. In the first chapter, a general review of breast cancer is carried out and microRNAs are also introduced. Next, in the second chapter, the objectives are detailed. In the third chapter, the microRNA expression profile of a doxorubicin-resistant breast cancer cell line was compared with its parental cell line, leading to the identification of the microRNA miR-99a-5p as the most deregulated between the two of them. Subsequently, it was confirmed that miR-99a-5p increases drug sensitivity through directly targeting COX-2, which leads to the inhibition of the expression of the ABCG2 protein, widely described to be involved in drug resistance. Based on the obtained results, nanodevices based on mesoporous silica nanoparticles were designed for the combined release of miR-99a-5p and doxorubicin in tumors. The ability of the systems to specifically target the CD44 receptor, which is overexpressed in breast cancer tumors, as well as to release the cargo specifically after internalization, were tested in vitro. Furthermore, the efficacy of the nanoparticles to reduce tumor growth and the adverse effects associated with free doxorubicin was confirmed in a murine orthotopic model of breast cancer. In the fourth chapter, the potential of miR-99a-5p as a biomarker for the diagnosis of breast cancer was evaluated. First, microRNA expression was determined in primary breast cancer tumors and compared with healthy tissues. The results showed that miR-99a-5p is downregulated in breast cancer tissues. Next, to determine the potential of the microRNA as a minimally invasive biomarker, the expression levels of miR-99a-5p in plasma of breast cancer patients and healthy donors were determined. In this case, it was found that miR-99a-5p is at higher concentrations in the plasma of breast cancer patients. These results were validated in an independent cohort. Through the ROC curve analysis, it was confirmed that miR-99a-5p is useful as a non-invasive biomarker for the diagnosis of breast cancer even at early stages. The fifth chapter focus on the design and validation of a biosensor based on nanoporous alumina supports for the detection of miR-99a-5p in plasma. The pores of the alumina plate were loaded with the fluorescent indicator rhodamine B, and oligonucleotides with a sequence complementary to the miR-99a-5p were used as molecular gates able to maintain fluorophores inside the pores. In the presence of miR-99a-5p, the capping oligonucleotide is displaced, leading to the opening of the pores and the release of rhodamine B, which is subsequently optically detected. The specificity and high sensitivity of the biosensor were confirmed. Furthermore, the efficiency of the device to detect breast cancer even at early stages was confirmed using plasma samples from breast cancer patients and healthy controls. Finally, chapters five and six present the general discussion and conclusions. In conclusion, this PhD thesis demonstrates that miR-99a-5p is a molecule with potential as a therapeutic target and biomarker for breast cancer, and it could be useful to improve the prognosis of patients. It has been elucidated that one of the mechanisms by which miR-99a-5p is involved in doxorubicin sensitivity is through the regulation of the COX-2/ABCG2 axis. Furthermore, nanoparticles designed for the combined administration of miR-99a-5p and doxorubicin are presented as a tool with great potential for cancer treatment and could be useful for the administration of microRNAs as therapy. Finally, taking advantage of the potential of miR-99a-5p as a diagnostic biomarker, a detection system has been developed and could be a useful tool to improve early detection of breast cancer. / This work was supported by, CIBERONC (CB16/12/00481), CIBER-BBN (CB07/01/2012), Spanish Government PI18/01219 (ISCIII), project RTI2018- 100910-B-C41 (MCUI/AEI/FEDER, UE) and the Generalitat Valenciana (PROMETEO/2018/024) for support. IG-C was funded by Generalitat Valenciana (ACIF/2016/030). LP was funded by Ministerio de Economia, Industria y Competitividad (FPI grant). SS was funded by Instituto de Salud Carlos III and the European Social Fund for the financial support “Sara Borrell” (CD16/000237). JMC was funded by Sociedad Española de Oncología Médica (Río Hortega-SEOM. The authors would like to thank the INCLIVA Biobank (PT17/0015/0049; B.000768 ISCIII) and the Valencian Biobanking Network integrated into the Spanish National Biobanks Network for providing the human biological samples. We would also like to thank the plasma donors and the nursing staff that accepted to participate in this study. / Garrido Cano, I. (2021). Role of miR-99a-5p in breast cancer: Translating molecular findings into clinical tools [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/180358

Female adults of many mosquito species possess distinct physiological features adapting to blood feeding for successful reproduction. The disease pathogens that are transmitted by mosquitoes have evolved to take advantages of the indispensable blood feedings to complete their transmission cycles and to survive attacks from the mosquito's innate immune system. Normal egg development and mosquito immunity are tightly controlled by tissue- and stage-specific gene expression and coordinated by many signal molecules in the mosquito. Understanding gene regulation affecting mosquito reproduction and malaria parasites infection is of paramount importance for developing novel malaria control strategies. A growing body of evidence indicates that microRNAs (miRNAs) are involved in egg maturation and immune reactions against invading pathogens in mosquitoes. However, the molecular mechanisms by which specific miRNAs selectively modulate reproduction and the survival of pathogens are largely unknown. The miRNA-induced gene-silencing pathway in mosquitoes was mostly extrapolated from the studies of flies. To explore the dynamics of miRNAs in reproduction, I used small RNAs sequencing to monitor miRNAs expression and their association with Argonaute 1 (Ago1) and Argonaute 2 (Ago2) in the malaria mosquito Anopheles gambiae (An. gambiae) during the 72-h period immediately after blood feeding. I found the abundance and Ago loading of most of the mature miRNAs were relatively stable after blood ingestion. However, miRNAs of the miR-309/286/2944 cluster were considerably upregulated after blood feeding. I confirmed that miR-309 is essential for normal egg development by depletion of endogenous miR-309 with a specific antagomir. In addition, my results showed that the Ago association of some miRNAs was not proportional to their cellular abundance implying additional regulation at miRNA integration. To investigate the functional roles of miRNAs and define context-dependent miRNA-mRNA interactions during the reproductive process, I have applied an innovative experimental approach to study miRNA-mRNA interactome. CLEAR (covalent ligation of endogenous Argonaute-bound RNAs)-CLIP can generate miRNA-mRNA chimeras from UV-irradiation stabilized Ago-miRNA-mRNA complex. My results have defined tens of thousands of miRNA-mRNA interactions in mosquitoes, including novel targets for mosquito-specific miRNAs. Verification of the predicted interactions using mRNA-seq, ribosome-profiling, and luciferase reporter assay revealed a reliable miRNA-mRNA interaction network. Based on the detected interactions, I refined the paring rules for mosquito miRNAs and illustrated the dynamic pairing between different regions of miRNAs with their targets in vivo. The miRNA-mRNA interactions were compared using this approach at multiple time points before and after blood feeding. Importantly, this study showed that the interactions were dynamic and enriched in genes that are involved in metabolisms, supporting the proposed functions of miRNAs in coordinating the gene regulation in mosquito reproduction. Plasmodium falciparum (P. falciparum) is a major human malaria parasite. To understand the functions of miRNAs in the mosquito resistance to Plasmodium infection, we analyzed the miRNA-mRNA interactions after female mosquitoes taking a P. falciparum-infected blood meal or an uninfected blood meal. Comparison of the interactions revealed enhanced miRNA-mRNA interactions after P. falciparum infection involving a group of immunity-related genes. In summary, this study has provided a systematic view and significantly advanced our understanding of the miRNA functions in mosquito reproduction and P. falciparum infection. / PHD / Female mosquito is able to transmit lots of disease to the human when it bites for blood. The blood meal provides necessary nutrient for mosquito reproduction and spread the pathogens such as malaria and Zika at the same time. Thus understanding the molecular mechanism behind this process would be greatly helpful to develop novel vector control strategy. Here, we found a distinct class of RNAs contributing to the regulation of mosquito blood meal and parasite infection. We used a novel biochemical method to decoding the special role of these kinds of RNAs in these processes. We found them regulating mosquito metabolism and immunity. This study significantly deepened our knowledge about the process of mosquito reproduction and transmitting diseases.

Introduction: In 2022 the conflict between Russia and Ukraine led Facebook to encourage a rise of hate speech and use of violence against Russian people. This led the Russian government to take measures and ban the use of western social media such as Facebook and Instagram in Russia and proclaim the Meta organization as an extremist organization. The ban caused various reactions which consequently led to a divergence in responses and opinions. Furthermore, the situation raises the question if the ban of western social media platforms is affecting Russian ICTs users' perception and political opinion. Aim: The aim of the study is to examine how the ban of western social media giants in Russia (Facebook, Instagram) affect Russian ICTs users’ attitude in the context of “critical citizen” or regime supporter. Also, the reasons why some Russians avoid western ICTs ban by using VPN and how some Russians experience ICTs censorship. A theoretical framework has been developed for the purpose to identify the different factors which might possibly lead to the emergence of “critical citizen” or pro-government citizen. Methodology: Semi-structured interviews were conducted to collect empirical data and fulfill the purpose of the thesis. Conclusion and contribution: The empirical data shows that the ban of Western ICTs (Facebook, Instagram) in Russia did not negatively affect ICTs users’ attitude toward Russian government nor encouraged an emergence of “critical citizen” rather in most cases raise an understanding and support of the ban even among those who still using Facebook and Instagram.  Findings show that VPN usage in most cases correlated with habits of preferences rather than a seeking of western political information. Furthermore, the user's perception on Facebook and Instagram censorship is more neutral/negative in context of general restriction and neutral/positive in context of Western influence resistance.

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No description available.

The present COVID-19 pandemic has revealed that several characteristics render patients especially prone to developing severe COVID-19 disease, i.e., the male sex, obesity, and old age. An explanation for the observed pattern of vulnerability has been proposed which is based on the concept of low sensitivity of the TLR7-signaling pathway at the time of infection as a common denominator of vulnerable patient groups. We will discuss whether the concept of established TLR-tolerance in macrophages and dendritic cells of the obese and elderly prior to infection can explain not only the vulnerability of these two demographic groups towards development of a severe infection with SARS-CoV-2, but also the observed cytokine response in these vulnerable patients, which is skewed towards pro-inflammatory cytokines with a missing interferon signature.