Efficacy of Infliximab after Failure of Subcutaneous Anti-TNF Agents in Patients with Moderate to Severe Ulcerative Colitis

Aim: To assess the efficacy of infliximab in ulcerative colitis (UC) patients who had failed therapy with adalimumab or golimumab. Methods: Retrospective analysis of prospectively acquired data of all anti-TNF naive patients with moderate to severe UC who received adalimumab or golimumab in 4 tertiary referral centres. Patients with primary non response or secondary loss of response to adalimumab or golimumab received therapy with infliximab. Clinical response and remission rates were assessed at week 14 and 54 after initiation of infliximab. Results: Between September 2015 and September 2017, 29 of 58 (50%) anti-TNF naive patients with moderate to severe UC failed therapy with adalimumab (n=38) or golimumab (n=20). Twenty one of 29 (72.4%) patients were primary non responders and 8 (27.6%) patients lost response to adalimumab or golimumab. All these 29 patients received infliximab, while 15 (51.7%) were on concomitant azathioprine therapy. Eighteen (62.1%) and 10 (34.5%) patients showed clinical response and clinical remission at week 14 respectively, while 14 (48.3%) patients were on clinical remission at week 54 after initiation of infliximab. Azathioprine co-administration at the start of infliximab was associated with a greater proportion of patients achieving clinical remission at week 54 (10 of 15 patients on combination therapy vs 4 of 14 patients on infliximab monotherapy, p=0.04). Conclusions: A significant proportion of anti-TNF naive patients with moderate to severe UC who have failed 1st course therapy with subcutaneous anti-TNF agents can achieve clinical response and/or remission with 2nd course therapy with infliximab

Risk of missing colorectal cancer with COVID-adapted diagnostic pathway using quantitative faecal immunochemical testing

Background: COVID-19 has brought an unprecedented challenge to healthcare services. The authors’ COVID-adapted pathway for suspected bowel cancer combines two quantitative faecal immunochemical tests (qFITs) with a standard CT scan with oral preparation (CT mini-prep). The aim of this study was to estimate the degree of risk mitigation and residual risk of undiagnosed colorectal cancer. Method: Decision-tree models were developed using a combination of data from the COVID-adapted pathway (April–May 2020), a local audit of qFIT for symptomatic patients performed since 2018, relevant data (prevalence of colorectal cancer and sensitivity and specificity of diagnostic tools) obtained from literature and a local cancer data set, and expert opinion for any missing data. The considered diagnostic scenarios included: single qFIT; two qFITs; single qFIT and CT mini-prep; two qFITs and CT mini-prep (enriched pathway). These were compared to the standard diagnostic pathway (colonoscopy or CT virtual colonoscopy (CTVC)). Results: The COVID-adapted pathway included 422 patients, whereas the audit of qFIT included more than 5000 patients. The risk of missing a colorectal cancer, if present, was estimated as high as 20.2 per cent with use of a single qFIT as a triage test. Using both a second qFIT and a CT mini-prep as add-on tests reduced the risk of missed cancer to 6.49 per cent. The trade-off was an increased rate of colonoscopy or CTVC, from 287 for a single qFIT to 418 for the double qFIT and CT mini-prep combination, per 1000 patients. Conclusion: Triage using qFIT alone could lead to a high rate of missed cancers. This may be reduced using CT mini-prep as an add-on test for triage to colonoscopy or CTVC

A molecular map of long non-coding RNA expression, isoform switching and alternative splicing in osteoarthritis

Osteoarthritis is a prevalent joint disease and a major cause of disability worldwide with no curative therapy. Development of disease-modifying therapies requires a better understanding of the molecular mechanisms underpinning disease. A hallmark of osteoarthritis is cartilage degradation. To define molecular events characterizing osteoarthritis at the whole transcriptome level, we performed deep RNA sequencing in paired samples of low- and high-osteoarthritis grade knee cartilage derived from 124 patients undergoing total joint replacement. We detected differential expression between low- and high-osteoarthritis grade articular cartilage for 365 genes and identified a 38-gene signature in osteoarthritis cartilage by replicating our findings in an independent dataset. We also found differential expression for 25 novel long non-coding RNA genes (lncRNAs) and identified potential lncRNA interactions with RNA-binding proteins in osteoarthritis. We assessed alterations in the relative usage of individual gene transcripts and identified differential transcript usage for 82 genes, including ABI3BP, coding for an extracellular matrix protein, AKT1S1, a negative regulator of the mTOR pathway and TPRM4, coding for a transient receptor potential channel. We further assessed genome-wide differential splicing, for the first time in osteoarthritis, and detected differential splicing for 209 genes, which were enriched for extracellular matrix, proteoglycans and integrin surface interactions terms. In the largest study of its kind in osteoarthritis, we find that isoform and splicing changes, in addition to extensive differences in both coding and non-coding sequence expression, are associated with disease and demonstrate a novel layer of genomic complexity to osteoarthritis pathogenesis

Primary osteoarthritis chondrocyte map of chromatin conformation reveals novel candidate effector genes

Objectives: Osteoarthritis is a complex disease with a huge public health burden. Genome-wide association studies (GWAS) have identified hundreds of osteoarthritis-associated sequence variants, but the effector genes underpinning these signals remain largely elusive. Understanding chromosome organisation in three-dimensional (3D) space is essential for identifying long-range contacts between distant genomic features (e.g., between genes and regulatory elements), in a tissue-specific manner. Here, we generate the first whole genome chromosome conformation analysis (Hi-C) map of primary osteoarthritis chondrocytes and identify novel candidate effector genes for the disease. Methods: Primary chondrocytes collected from 8 patients with knee osteoarthritis underwent Hi-C analysis to link chromosomal structure to genomic sequence. The identified loops were then combined with osteoarthritis GWAS results and epigenomic data from primary knee osteoarthritis chondrocytes to identify variants involved in gene regulation via enhancer-promoter interactions. Results: We identified 345 genetic variants residing within chromatin loop anchors that are associated with 77 osteoarthritis GWAS signals. Ten of these variants reside directly in enhancer regions of 10 newly described active enhancer-promoter loops, identified with multiomics analysis of publicly available chromatin immunoprecipitation sequencing (ChIP-seq) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) data from primary knee chondrocyte cells, pointing to two new candidate effector genes SPRY4 and PAPPA (pregnancy-associated plasma protein A) as well as further support for the gene SLC44A2 known to be involved in osteoarthritis. For example, PAPPA is directly associated with the turnover of insulin-like growth factor 1 (IGF-1) proteins, and IGF-1 is an important factor in the repair of damaged chondrocytes. Conclusions: We have constructed the first Hi-C map of primary human chondrocytes and have made it available as a resource for the scientific community. By integrating 3D genomics with large-scale genetic association and epigenetic data, we identify novel candidate effector genes for osteoarthritis, which enhance our understanding of disease and can serve as putative high-value novel drug targets

Volatile organic compounds in breath can serve as a non-invasive diagnostic biomarker for the detection of advanced adenomas and colorectal cancer

Contains fulltext : 220031.pdf (Publisher’s version ) (Open Access)BACKGROUND: Colorectal cancer (CRC) is the third most common cancer diagnosis in the Western world. AIM: To evaluate exhaled volatile organic compounds (VOCs) as a non-invasive biomarker for the detection of CRC and precursor lesions using an electronic nose. METHODS: In this multicentre study adult colonoscopy patients, without inflammatory bowel disease or (previous) malignancy, were invited for breath analysis. Two-thirds of the breath tests were randomly assigned to develop training models which were used to predict the diagnosis of the remaining patients (external validation). In the end, all data were used to develop final-disease models to further improve the discriminatory power of the algorithms. RESULTS: Five hundred and eleven breath samples were collected. Sixty-four patients were excluded due to an inadequate breath test (n = 51), incomplete colonoscopy (n = 8) or colitis (n = 5). Classification was based on the most advanced lesion found; CRC (n = 70), advanced adenomas (AAs) (n = 117), non-advanced adenoma (n = 117), hyperplastic polyp (n = 15), normal colonoscopy (n = 125). Training models for CRC and AAs had an area under the curve (AUC) of 0.76 and 0.71 and blind validation resulted in an AUC of 0.74 and 0.61 respectively. Final models for CRC and AAs yielded an AUC of 0.84 (sensitivity 95% and specificity 64%) and 0.73 (sensitivity and specificity 79% and 59%) respectively. CONCLUSIONS: This study suggests that exhaled VOCs could potentially serve as a non-invasive biomarker for the detection of CRC and AAs. Future studies including more patients could further improve the discriminatory potential of VOC analysis for the detection of (pre-)malignant colorectal lesions. (https://clinicaltrials.gov Identifier NCT03488537)

Development of a novel thermal insulation system for building envelope application

The building sector is responsible for 38% of global energy and process-related greenhouse gas emissions. In recent decades, a rapid increase in energy consumption in buildings has been witnessed, making energy reduction a pressing issue. Thermal insulation systems, that can operate dynamically in response to changing transient conditions, by alternating between thermally conductive and insulated states, are gaining attention in their architectural applications, since they constitute an effective mean for reducing energy consumption while simultaneously improving occupants comfort levels. Despite the pioneering work that has been conducted by researchers to develop adaptive insulation technologies in different engineering fields, currently none of the adaptive insulation technologies are embedded in real-world buildings for a number of reasons. The aim of this thesis, therefore is to identify these reasons and on this basis, to explore, the different ways in which an adaptive insulation system can be created. There are no established standards or requirements for the development of adaptive thermal insulation technologies. Consequently, this research initially focuses on the creation of design criteria that address aspects related to thermal performance, design feasibility, and technological complexity of the examined systems. Subsequently, the exploration of different design concepts was initiated by first creating a scheme of aspects, concerning the geometry of the core, the mobility of the outer layer, the number of cavity compartments, the type of actuation and the types of mechanisms that facilitate the transition between thermal insulation states. From the combination of these aspects, six design alternatives emerged, the principle of operation of which is based on the idea of a structure that can inflate and deflate during the transition of the system from the insulated to the conductive state. The core geometry of the concepts was modeled within TRISCO steady-state 3D software tool, where various parameters were tested in order to obtain the final topology. The aim was to achieve a thermal resistance comparable to that of state-of-the-art insulating materials in their insulated state and furthermore, to achieve a large range of shift in order to acquire a significantly low thermal resistance in the conductive state comparable to the case of an uninsulated concrete block. Simultaneously, the thermal transmittance (U-value) for the insulated and conductive states of each concept was estimated through numerical simulations in TRISCO and validated using analytical models for comparison. The results of the analysis indicated that, the thickness of the air cavity and the emissivity of the membrane material greatly affect the examined thermal resistance. Additionally, the range of shift depends significantly on the degree of evacuation of the air cavity. It has been found that technologies whose structure and working principle allow full compression of the core in the conductive state, can lead to a lower thermal resistance which is governed by solid conduction. The results of the simulations showed that one of the examined concepts satisfied the initial goal, achieving thermal transmission which ranged from 0.13 W/𝑚2K to 2.79 W/𝑚2K in the insulated and conductive states respectively. Whereas, the thermal resistance ranged from 7.5 𝑚2/KW to 0.4 𝑚2/KW in the insulatedand conductive states respectively. The obtained values of thermal transmittance from TRISCO, were compared with the thermal transmittance from analytical formulas. In this way, the accuracy of the numerical simulations was validated. The final stage of the design exploration phase led to the selection of the final design through a multi-criteria analysis that is aligned with the aforementioned initial design criteria. The final design is a 1.5 m by 0.75 m opaque panel of low emissivity honeycomb core made up of 10 mm cells. In the insulated state the panel has a thickness of 0.18 m while after being compressed the thickness is reduced to 1/3. Actuation of the system is achieved using a dual-function pump that supplies air to a system of channels that access the core. An approximation of the sizes of the pump and pipes were given for the design of the air-supply system. In this way, the system can be reversibly switched between an inflated state governed by gaseous conduction and a deflated state, where the solid conduction of adjacent surfaces governs. The architectural application of the adaptive insulation system, considers the element as an infill component in which the outer skin is integrated into the system, implying that the transition between thermal states is visible from the façade. To conclude, this research explores the potentials of using the parameters influencing heat transfer capability for the development of an adaptive insulation system intended to be used in the building envelope. The research through design resulted in a promising design based on the resulting thermal performance indicators. However, to realize the proposed technology, tests on physical model need to be conducted. The thermal performance of the system should be validated experimentally in order to derive statistical data for its safe application and the actual dimensions of the air-supply system need to be determined.Architecture, Urbanism and Building Sciences | Building Technolog

IoT ekosistemų panaudojimas sporto vartotojų elgsenos monetizavimui

The emergence of technology and IoT, has resulted in the creation of a complex and multipurpose usage of consumers engagement behavior data. Nowadays, sports consumers function in complex ecosystems that comprise by many different stakeholders who must work together to provide the best services and unlock the commercial potential of the so-called IoT potential. By connecting devices, vehicles, infrastructures and cities with people, businesses can generate new revenue streams and other economic and financial aspects. This fact leads to a twofold contribution of this paper: (i) identify and quantify the sports consumer behavior (ii) propose a monetization model in that will evolve sports consumers data experience within IoT ecosystem. Within this context, the practicability of this relationship between the sports consumer behavior and IoT applications, along with a sports consumer evolution analysis within this context are analyzed considering a proof-of-concept for helping businesses and organizations that offer IoT-enabled products or services, to most value from their content

Linguistic and stylistic approach on the Plutarch's Moralia

The present doctoral thesis examines how the multifaceted personality of Plutarch’s is reflected lexicographically and stylistically in the corpus of Moralia and more specifically highlights the following aspects: the didactic, the scientific, the religious and the collaborative. With reference to the lexicological part of the research, linguistic terms are presented that are connected to the traditional vocabulary of religion and science, which the prose writer used in his work, but where the main emphasis is on the analysis of the linguistic creations of the Chaironian writer, which are related with the specific sectors. Issues related to the construction of the philosopher’s neologisms are also investigated and one of them refers to his preference to proceed with the creation of terms, which carry as their first linguistic garments some specific intentions, such as the uninflected word σύν, because with its help managed to verbally capture a basic concept that resided in his thought, that of collectivity. The stylistic observations of this particular study concern the world of impersonal verbs and verbal adjectives in -τέον. Of all the impersonal verbs the prose writer used, with greater intensity and frequency, δεῖ and within the doctoral thesis are analyzed and explained the reasons which prompted him to make this specific stylistic choice, as the various concepts with connected δεῖ as well as some more impersonal verbs, such as χρή and προσήκει. In addition to impersonal verbs Plutarch in Moralia made extensive use of the verbal adjectives in -τέον in order to convey his moral teachings.Η παρούσα διατριβή εξετάζει πώς αποτυπώνεται λεξικογραφικά και υφολογικά η πολυσχιδής προσωπικότητα του Πλουτάρχου στο corpus των Ηθικών και πιο συγκεκριμένα αναδεικνύει τις ακόλουθες πτυχές της: τη διδακτική, την επιστημονική, τη θρησκευτική και τη συλλογική. Αναφορικά με το λεξικολογικό τμήμα της έρευνας παρουσιάζονται γλωσσικοί όροι που συνδέονται με το παραδοσιακό λεξιλόγιο της θρησκείας και της επιστήμης, το οποίο χρησιμοποίησε ο πεζογράφος στην εργογραφία του, αλλά εκεί όπου δίνεται κυρίως έμφαση είναι στην ανάλυση των ἅπαξ λεγομένων του χαιρώνειου συγγραφέα, τα οποία σχετίζονται με τους συγκεκριμένους τομείς. Διερευνώνται και θέματα σχετικά με την κατασκευή των νεολογισμών του φιλοσόφου και ένα από αυτά αναφέρεται στην προτίμησή του να προβαίνει στη λεξιπλασία όρων, οι οποίοι φέρουν ως πρώτα γλωσσικά τους ενδύματα κάποιες συγκεκριμένες προθέσεις, όπως είναι η άκλιτη λέξη σύν, διότι με τη βοήθειά της κατάφερε να αποτυπώσει λεκτικά μια βασική έννοια που κατοικούσε στη σκέψη του, αυτήν της συλλογικότητας. Οι υφολογικές παρατηρήσεις της συγκεκριμένης μελέτης αφορούν τον κόσμο των απρόσωπων ρημάτων και των ρηματικών επιθέτων σε -τέον. Από όλα τα απρόσωπα ρήματα ο πεζογράφος χρησιμοποίησε, με μεγαλύτερη ένταση και συχνότητα, το δεῖ και εντός της διατριβής αναλύονται και επεξηγούνται οι λόγοι, οι οποίοι τον ώθησαν στη συγκεκριμένη υφολογική επιλογή καθώς παρουσιάζονται και οι διάφορες έννοιες με τις οποίες συνέδεσε το δεῖ, αλλά και ορισμένα ακόμα απρόσωπα ρήματα, όπως το χρή και το προσήκει. Πέρα από τα απρόσωπα ρήματα ο Πλούταρχος στα Ηθικά προέβη και σε διευρυμένη χρήση και των ρηματικών επιθέτων σε -τέον προκειμένου να μεταλαμπαδεύσει με μεγαλύτερη ευκρίνεια τα νάματα αρετής και παιδείας του

Μελέτη της λειτουργίας και ποικιλότητας της μικροβιακής κοινότητας στην φυλλόσφαιρα και ριζόσφαιρα φυτών και αλληλεπιδράσεις με το περιβάλλον

Phyllosphere and rhizosphere are plant-associated micro-habitats that are known to support diverse microbial communities whose structure is mediated by plants. We aimed to disentangle the mechanisms shaping the microbial communities in the phyllosphere and the soil root zone and identify their response to agricultural practices like soil organic amendment and pesticide application. The focus was on plants indigenous to Mediterranean ecosystems, some of them producing essential oils which are known to exert antimicrobial activities, and also cultivated plants. We initially explored the factors shaping the microbial community of the phyllosphere in plants native to semi-arid Mediterranean ecosystems using q-PCR and amplicon sequencing approaches. We collected leaves at two largely contrasting seasons (summer and winter) from 8 perennial plants with varying attributes, that belong to different functional groups: (i) woody sclerophyllous evergreen, semi-deciduous and non woody shrubs (ii) aromatic and non-aromatic. We determined the abundance of bacteria, Crenarchaea, fungi, Alternaria and Cladosporium (main airborne fungi) via q-PCR, and the structure of the epiphytic bacterial, archaeal and fungal community via amplicon sequencing. We observed strong seasonal effects but no clear plant-host effects on microbial abundance: bacteria showing higher abundance in the winter, and all others in the summer. Plant-host and season were equal determinants of the composition of the bacterial and fungal communities, whereas the archaeal community showed plant-host driven patterns. Plant habit exhibited a stronger filtering effect on the epiphytic microbial communities compared to the aromatic plant nature which affected only the fungal community. The bacterial community was dominated by Chloroflexi and α-proteobacteria in the summer and winter respectively, with OTUs of Sphingomonas, Rhizobia and Methylobacterium favored in the winter. The archaeal community was dominated by the Soil Crenarchaeotic Group (SCG) and Aenigmarchaeota. The fungal community is mostly comprised of Ascomycota with Capnodiales, Pleosporaceae and Dothioraceae being key members whose abundance varied by plant host and season. We extended our study on aromatic plants by exploring their use, as soil amendments. We employed a pot study to examine the impact of peppermint (Menta piperita), spearmint (Menta spicata) and rosemary (Rosemarinus officinalis), in comparison with an organic fertilizer, on the dynamics of key bacterial taxa, Crenarchaea, fungi and functional microbial groups like ammonia-oxidizing bacteria (AOB) and archaea (AOA), sulfur-oxidizing bacteria (SOB) and catA-, pcaH-carrying bacteria involved in C cycling. We further explored possible interactions between soil amendments and the presence of tomato plant. Soil amendment with peppermint, spearmint and the organic fertilizer increased the abundance of proteobacteria and fungi, in contrast to rosemary, characterized by essential oils with a different chemical profile compared to mints, which benefited these copiotrophic microbial groups only in the presence of tomato plants. We further explored this complex interaction via amplicon sequencing analysis of bacteria, archaea and fungi. This verified the key role of rosemary soil amendment in shaping the bacterial, archaeal and fungal community and its beneficial role in the abundance of proteobacteria. Multivariate analysis identified OTUs belonging to Actinobacteria, mostly associated with undisturbed soil systems (i.e Blastococcus, Rubrobacter, Solirubrobacter, Agromyces) that were negatively affected by rosemary amendment. On the contrary we observed a striking dominance of the cellulose-decomposing basidiomycetes Minimedusa in soils amended with rosemary. The known antibiotic properties of this fungus might explain the negative effects of rosemary soil amendment on Nectriaceae also observed. We finally explored the potential impact of pesticides, as external perturbation factor, on the abundance and diversity of the microbial communities on plant leaves and the soil root zone. We tested the hypothesis that these two habitats support distinct microbial communities but exhibit a similar response (accelerated biodegradation or toxicity) to their repeated exposure to the biodegradable fungicide iprodione. Pepper plants received four repeated foliage or soil applications of iprodione which accelerated its degradation in soil (DT50_1st=1.23 and DT50_4th = 0.48 days) and on plant leaves (DT50_1st >365 and DT50_4th = 5.95 days). The composition of the epiphytic and soil bacterial and fungal communities, determined by amplicon sequencing, were significantly altered by iprodione. The archaeal epiphytic and soil communities responded differently; the former showed no response to iprodione. Three closely related iprodione-degrading Paenarthrobacter strains were isolated from soil and phyllosphere. They hydrolyzed iprodione to 3,5-dichloraniline (3,5-DCA) via the formation of 3,5-dichlorophenyl-carboxiamide and 3,5-dichlorophenylurea-acetate, a pathway shared by other soil-derived arthrobacters implying a phylogenetic specialization in iprodione biotransformation. Overall, we showed that phyllosphere is a habitat colonized by diverse bacteria and fungi, while archaea are less abundant and diverse. The epiphytic microbial community in Mediterranean plants, is shaped by plant-host and seasonality. The use of aromatic plants as soil amendment was found to stimulate copiotrophic microorganisms and microorganisms allelopathic against soil-borne plant pathogens. Finally, we showed that the epiphytic microbiome, responds to pesticide applications, with some microbes became acclimated to degrade pesticides. This thesis has reported the first epiphytic bacterium, a Paenarthrobacter strain, that could degrade iprodione and also suggested an uncommon specialization of Arthrobacter in the degradation of this fungicide.Η φυλλόσφαιρα και η ριζόσφαιρα είναι μικρο-ενδιαιτήματα του φυτού που είναι γνωστό ότι φέρουν ποικιλόμορφες μικροβιακές κοινότητες, η δομή των οποίων εξαρτάται από το φυτό. Σκοπός μας ήταν η διερεύνηση των μηχανισμών που διαμορφώνουν τη μικροβιακή κοινότητα της φυλλόσφαιρας και του εδάφους που βρίσκεται περιμετρικά της ρίζας, και η ταυτοποίηση της απόκρισης τους σε καλλιεργητικές πρακτικές, όπως η χρήση εδαφοβελτιωτικών και οι εφαρμογές γεωργικών φαρμάκων. Εστιάσαμε σε γηγενή φυτά του Μεσογειακού συστήματος, μερικά εκ των οποίων παράγουν αιθέρια έλαια που είναι γνωστά για τις αντιμικροβιακές τους ιδιότητες, και επίσης σε καλλιεργούμενα φυτά. Αρχικά διερευνήσαμε τους παράγοντες που διαμορφώνουν τη μικροβιακή κοινότητα της φυλλόσφαιρας γηγενών φυτών του ημιάνυδρου Μεσογειακού οικοσυστήματος, χρησιμοποιώντας προσεγγίσεις με q-PCR και amplicon sequencing. Συλλέξαμε φύλλα στις δύο εποχές με τις μεγαλύτερες αντιθέσεις (καλοκαίρι και χειμώνας) από 8 πολυετή φυτά με διαφορετικά χαρακτηριστικά που ανήκουν σε διάφορες λειτουργικές ομάδες: (i) αειθαλή ξυλώδη σκληρόφυλλα, φρύγανα και πόες, (ii) αρωματικά και μη-αρωματικά. Καθορίσαμε την αφθονία των βακτηρίων, Κρεναρχαίων, μυκήτων Alternaria και Cladosporium (κύριοι αερομεταφερόμενοι μύκητες) μέσω q-PCR, και τη δομή της επιφυτικής κοινότητας των βακτηρίων, αρχαίων και μυκήτων μέσω amplicon sequencing. Παρατηρήσαμε έντονη επίδραση της εποχής, αλλά όχι ξεκάθαρη επίδραση του φυτού-ξενιστή στη μικροβιακή αφθονία: η βακτηριακή αφθονία ήταν υψηλότερη τον χειμώνα, και η αφθονία όλων των υπόλοιπων μικροοργανισμών ήταν αυξημένη το καλοκαίρι. Φυτό-ξενιστής και εποχή είχαν ίση επίδραση στη σύσταση της μικροβιακής κοινότητας των βακτηρίων και μυκήτων, ενώ η κοινότητα των αρχαίων καθορίζεται από το φυτό-ξενιστή. Το φυτικό είδος αποτελεί σημαντικότερο παράγοντα διαμόρφωσης των επιφυτικών μικροβιακών κοινοτήτων, σε σχέση με την αρωματική φύση των φυτών, η οποία επηρεάζει μόνο τη μυκητιακή κοινότητα. Η βακτηριακή κοινότητα κυριαρχείται από Chloroflexi και α-Πρωτεοβακτήρια το καλοκαίρι και χειμώνα αντίστοιχα, με τα OTUs των Sphingomonas, Rhizobia και Methylobacterium να ευνοούνται τον χειμώνα. Η κοινότητα των αρχαίων κυριαρχείται από Soil Crenarchaeotic Group (SCG) και Aenigmarchaeota. Η μυκητιακή κοινότητα αποτελείται κυρίως από Ascomycota με Capnodiales, Pleosporaceae και Dothioraceae, να είναι τα μέλη-κλειδιά των οποίων η αφθονία διαφοροποιείται αναλόγως του φυτού-ξενιστή και της εποχής. Επεκτείναμε τη μελέτη μας και στα αρωματικά φυτά, εξερευνώντας τη χρήση τους ως εδαφοβελτιωτικά. Πραγματοποιήσαμε ένα πείραμα με γλάστρες, ώστε να μελετήσουμε την επίδραση της μέντας (Menta piperita), του δυόσμου (Menta spicata) και του δενδρολίβανου (Rosemarinus officinalis), σε σύγκριση με ένα οργανικό εδαφοβελτιωτικό, στη δυναμική των κύριων βακτηριακών taxa, Κρεναρχαίων, μυκήτων και άλλων λειτουργικών μικροβιακών ομάδων, όπως τα αμμώνια-οξειδωτικά βακτήρια (AOB) και αρχαία (AOA), τα θείο-οξειδωτικά βακτήρια (SOB) και τα catA-, pcaH- φέροντα βακτήρια, τα οποία εμπλέκονται στον κύκλο του άνθρακα. Επιπλέον διερευνήσαμε πιθανές αλληλεπιδράσεις μεταξύ εδαφοβελτιωτικών και παρουσίας του φυτού τομάτας. Η βελτίωση του εδάφους με μέντα, δυόσμο και οργανικό εδαφοβελτιωτικό, αύξησε την αφθονία των Πρωτεοβακτηρίων και μυκήτων. Αντίθετα το δενδρολίβανο, το οποίο έχει χαρακτηριστικό αιθέριο έλαιο με διαφορετική σύσταση σε σχέση με τις μέντες έδρασε ευεργετικά στις κοπιοτροφικές αυτές μικροβιακές ομάδες, μόνο κατά την παρουσία του φυτού της τομάτας. Επιπροσθέτως εξερευνήσαμε τις πολύπλοκες αλληλεπιδράσεις μέσω ανάλυσης amplicon sequencing στις μικροβιακές κοινότητες των βακτηρίων, αρχαίων και μυκήτων. Αυτό επιβεβαίωσε τον ρόλο κλειδί του δενδρολίβανου ως εδαφοβελτιωτικού στη σύσταση της βακτηριακής και μυκητιακής κοινότητας καθώς και αυτής των αρχαίων, αλλά και τον ευεργετικό του ρόλο στην αφθονία των Πρωτεοβακτηρίων. Η εφαρμογή του δενδρολίβανου επηρέασε αρνητικά OTUs που ανήκουν στα Ακτινοβακτήρια, τα οποία σχετίζονται κυρίως με ανέγγιχτα εδαφικά συστήματα (π.χ. Blastococcus, Rubrobacter, Solirubrobacter, Agromyces), όπως προέκυψε από την πολυπαραγοντική ανάλυση. Αντίθετα παρατηρήθηκε μία εντυπωσιακή κυριαρχία των διασπαστών κυτταρίνης βασιδιομυκήτων Minimedusa σε εδάφη εμπλουτισμένα με δενδρολίβανο. Οι γνωστές αντιμικροβιακές ιδιότητες του μύκητα αυτού ίσως να εξηγούν την αρνητική επιρροή του δενδρολίβανου ως εδαφοβελτιωτικού, που παρατηρήθηκε στα Nectriaceae. Τέλος, εξερευνήσαμε την πιθανή επίδραση των γεωργικών φαρμάκων, ως εξωτερικού παράγοντα διαταραχής της αφθονίας και ποικιλότητας των μικροβιακών κοινοτήτων στα φύλλα του φυτού και στο έδαφος στην περιοχή της ρίζας. Εξετάσαμε την υπόθεση ότι αυτά τα δύο ενδιαιτήματα ενώ φέρουν διαφορετικές μικροβιακές κοινότητες, αντιδρούν το ίδιο (επιταχυνόμενη βιοδιάσπαση ή τοξικότητα) στην επαναλαμβανόμενη έκθεση τους στο βιοδιασπώμενο μυκητοκτόνο iprodione. Σε φυτά πιπεριάς εφαρμόστηκαν τέσσερις επαναλαμβανόμενες φυλλικές ή εδαφικές εφαρμογές του iprodione, διαδικασία που επιτάχυνε τη διάσπαση στο έδαφος (DT50_1st=1.23 και DT50_4th = 0.48 ημέρες) και στα φύλλα (DT50_1st >365 και DT50_4th = 5.95 ημέρες). Η σύσταση της επιφυτικής και εδαφικής βακτηριακής και μυκητιακής κοινότητας, η οποία καθορίστηκε με amplicon sequencing, άλλαξε στατιστικώς σημαντικά από το iprodione. Η επιφυτική και εδαφική κοινότητα των αρχαίων αντέδρασε διαφορετικά· με το πρώτο να μην αντιδρά στο iprodione. Τρία συγγενικά στελέχη Paenarthrobacter που διασπούν το iprodione, απομονώθηκαν από έδαφος και φύλλα. Υδρολύουν το iprodione σε 3,5-dichloraniline (3,5-DCA) μέσω σχηματισμού 3,5-dichlorophenyl-carboxiamide και 3,5-dichlorophenylurea-acetate, ένα μονοπάτι που μοιράζονται με άλλα arthrobacters που έχουν απομονωθεί από το έδαφος, προτείνοντας μία φυλογενετική ειδίκευση στη βιομετασχηματισμό του iprodione. Συνολικά, αποδείξαμε πως η φυλλόσφαιρα είναι ένα ενδιαίτημα που αποικίζεται από διαφορετικά βακτήρια και μύκητες, ενώ η αφθονία και ποικιλομορφία των αρχαίων είναι μειωμένη. Η επιφυτική μικροβιακή κοινότητα των Μεσογειακών φυτών διαμορφώνεται από το φυτό-ξενιστή και την εποχή. Η εφαρμογή αρωματικών φυτών ως εδαφοβελτιωτικού βρέθηκε να διεγείρει τους κοπιοτροφικούς μικροοργανισμούς και αλληλοπαθητικούς εναντίων εδαφογενών φυτοπαθογόνων. Τέλος, αποδείξαμε ότι το επιφυτικό μικροβίωμα, αντιδρά στις εφαρμογές γεωργικών φαρμάκων, με μερικά μικρόβια να εγκλιματίζονται στο να διασπούν γεωργικά φάρμακα. Αυτή η διδακτορική διατριβή ανέφερε το πρώτο επιφυτικό βακτήριο, ένα στέλεχος Paenarthrobacter, το οποίο είναι ικανό να διασπά το iprodione και επίσης προτείνει μία μη-κοινή ειδίκευση των Arthrobacter στη διάσπαση αυτού του μυκητοκτόνου