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Siklamende görülen kendileme depresyonu, farklı ploidi seviyeleri, kendine uyuşmazlık ve abortifembriyo oluşumu nedeniyle klasik yöntemlerle siklamen ıslahı oldukça zordur. Bu nedenle in vitrohaploidizasyon, siklamen ıslahında yaygın olarak kullanılmaktadır. Bu çalışmada in vitrohaploidizasyon ve in vitro çoğaltımın ilk aşaması olarak bitki doku kültürü ortamlarının ve hormonlarınetkisi araştırılmıştır. Çalışmada bitkisel materyal olarak, Cyclamen persicum ve Cyclamenpseudibericum’un ovül eksplantları, ortam olarak ise 2,4-dichlorophenoxyaceticacid (2,4-D) ve 6-(?,? –dimethylallylamino) purine’ (2iP)’in farklı konsantrasyonlarını içeren ½MS ve B5 ortamlarıkullanılmıştır. Çalışma sonucunda, Cyclamen persicum türünde en iyi kallus oluşum oranı; 1.5 mg L-1(2,4-D ve 0.5 mg L-12iP içeren B5 ortamından (%70 oranında) elde edilmiştir. Cyclamenpseudibericum türü için ise kallus oluşum oranı % 48 olmuş ve 1 mg L-12,4-D ve 0.3 mg L-12iP içerenB5 ortamından elde edilmiştir. Cyclamen persicum türünde en yüksek embriyo uyartımı 1.5 mg L-12,4-D ve 0.8 mg L-12iP içeren ½MS ortamında gelişen kalluslardan elde edilmiştir (%55). Ancak Cyclamenpseudibericum türünde besi ortamlarının hiç birinden embriyo uyartımı sağlanamamıştır. Sonuç olarak,kallus uyartımında B5 ortamı, ½MS ortamından daha iyi sonuç vermesine rağmen embriyo uyartımıiçin yüksek oksin ve düşük sitokinin konsatrasyonuna sahip ½MS ortamı daha iyi bulunmuş ve 2.0 mgL-12,4-D ve 0.5 mg L-12iP içeren ½MS ortamından 42 adet bitkicik elde edilmiştir. Cyclamen breeding is difficult due to selfing depression different ploidy levels, self-incompatibility andabortive embryo formation. Therefore, in vitro haploidisation is commonly used in cyclamen breeding.In this study, we investigated effects of different tissue culture media and plant hormones as a first of invitro haploidisation and propagation. We used ovule explants from Cyclamen persicum and Cyclamenpseudibericum on ½MS and B5 culture mediums supplemented with different concentration of 2,4-dichlorophenoxyaceticacid (2,4-D) and 6-(?,? –dimethylallylamino) purine (2iP). According to resultsbest callus formation rate was 70% obtained from B5 medium supplemented with 1.5 mg L-12,4-D and0.5 mg L-1 2iP in Cyclamen persicum. It was 48% on B5 medium supplemented with 1 mg L-12,4-Dand 0.3 mg L-12iP in Cyclamen pseudibericum. In Cyclamen persicum, highest embryo induction ratewas 55% obtained from callus regenerated on ½MS medium supplemented with 1.5 mg L-12,4-D and0.8 mg L-12iP. However, embryo induction was not obtained on any medium for Cyclamenpseudibericum. Overall, ½MS culture medium supplemented with high auxin and low cytokininconcentration is better for embryo induction although B5 medium was better than ½MS medium forcallus formation and 42 plantlets obtained only from callus regenerated on ½MS medium supplementedwith 2.0 mg L-12,4-D and 0.5 mg L-1 2iP.

Arbuscular mycorrhizal symbiosis is a mutualistic association established between the roots of most of land plants and soil fungi of the Glomeromycete subphylum. This interaction is beneficial for both partners: the fungus provides the plant with minerals (mainly phosphorus and nitrogen) and in return obtains carbon molecules (in the form of lipids and oses), which are essential for its survival. This interaction is initiated by a molecular dialogue, involving chitinic molecules from fungal origin and phytohormones secreted by the plant, the strigolactones. Once the symbiosis is established, it is controlled by plant hormones, which promote or inhibit its maintenance. Interestingly, although many soil bacterial and fungal species have been able to produce plant hormones to modulate their interactions with plants, few studies describe the ability of arbuscular mycorrhizal fungi to biosynthesize such molecules. Furthermore, genes encoding two putative phytohormone receptors - Cytokinins and Ethylene - have however been identified in certain endomycorrhizal fungi. The first objective of this work was to identify plant hormones in the model endomycorrhizal fungus Rhizophagus irregularis. As this fungus is an obligate biotroph, unable to develop in the absence of a plant partner, we used Rhizophagus irregularis germinating spores in axenic conditions in order to avoid any contamination from the plant. Using sensitive and precise biochemical analysis methods, we have demonstrated the presence of four phytohormones in fungal extracts: an auxin (indole-3-acetic acid), a cytokinin (isopentenyl-adenosine), a gibberellic acid (GA4) and ethylene. Our experiments further suggest that, like in most fungi, R. irregularis uses a KMBA-dependent pathway to biosynthesize ethylene. The second objective was to determine the origin of biosynthesis of these hormones. They can either be metabolized by the fungus, or can be imported during symbiotic exchanges with the plant. To demonstrate a possible fungal biosynthesis, we initiated isotopic labelling experiments, by germinating spores in the presence of labelled carbon sources. A third objective was to initiate work on the effects of cytokinins and ethylene on R. irregularis, since it has putative receptors for these two phytohormones. Biochemical evidence for the presence of hormones in R. irregularis is an important discovery for the community. This discovery opens up new perspectives to understand their roles and regulations during the symbiosis. These could have a direct impact on the physiology of endomycorrhizal fungi, contribute to the plant-fungus or inter-plant molecular dialogue, via the mycelial network, but also influence the microbial communities in the soil. These results will further encourage the study of plants in their natural microbiome and will provide a better understanding of how this common hormonal language, has contributed to this symbiosis through the evolution, that started when plants colonized the land and interacted with soil fungi.; La symbiose mycorhizienne à arbuscules est une association mutualiste qui s'établit entre les racines de la majorité des plantes terrestres et les champignons souterrains du sous-phylum des Gloméromycetes. Cette interaction est bénéfique pour les deux partenaires : le champignon fournit à la plante des minéraux (principalement phosphore et azote) et il obtient en retour des molécules carbonées (sous forme de lipides et d'oses), indispensables à sa survie. Cette interaction est initiée par un dialogue moléculaire, impliquant d'une part des molécules chitiniques d'origine fongique et d'autre part des hormones secrétées par la plante, les strigolactones. Une fois la symbiose établie, elle est contrôlée par des hormones végétales, qui favorisent ou inhibent son maintien. De manière intéressante, malgré que de nombreuses espèces bactériennes et fongiques du sol aient été décrites comme capables de produire des hormones végétales, pour moduler leurs interactions avec les plantes, peu d'études décrivent la capacité des champignons mycorhiziens à arbuscules à biosynthétiser de telles molécules. Pourtant, des gènes codants pour des récepteurs putatifs à deux phytohormones - Cytokinines et Ethylène - ont été identifiés chez certains champignons endomycorhiziens. Le premier objectif de ce travail a été d'identifier des hormones végétales chez le champignon endomycorhizien modèle Rhizophagus irregularis. Ce champignon étant biotrophe obligatoire, incapable de se développer en absence de partenaire végétal, nous avons utilisé des spores en germination axénique afin d'éviter toute contamination issue de la plante. A l'aide de méthodes d'analyses biochimiques, sensibles et précises, nous avons mis en évidence la présence de quatre hormones végétales dans les extraits fongiques : une auxine (acide indole-3-acétique), une cytokinine (isopentenyl-adénosine), un acide gibbérellique (GA_4) et l'éthylène. Nos expériences suggèrent, de plus, que comme chez la plupart des champignons, R. irregularis utilise une voie KMBA-dépendante pour biosynthétiser l'ethylène. Le deuxième objectif a été de déterminer l'origine de biosynthèse de ces hormones. Elles peuvent soit être métabolisées par le champignon, soit être importées durant les échanges symbiotiques avec la plante. Pour démontrer une éventuelle biosynthèse fongique, nous avons initié des expériences d'incorporation et de suivi de marquages isotopiques, en faisant germer des spores en présence de sources de carbones marqués. Un troisième objectif a visé à initier des travaux sur les effets des cytokinines et de l'éthylène sur R. irregularis, puisqu'il possède des récepteurs putatifs à ces deux phytohormones. La preuve biochimique de la présence d'hormones chez R. irregularis constitue une découverte importante pour la communauté. Cette découverte ouvre de nouvelles perspectives quant à leurs rôles et leurs régulations lors de la symbiose. Celles-ci pourraient avoir un impact direct sur la physiologie des champignons endomycorhiziens, contribuer au dialogue moléculaire plante-champignon ou inter-plantes, via le réseau mycélien, mais également influencer les communautés microbiennes du sol. Ces résultats encourageront à davantage étudier les plantes dans leur microbiome naturel et permettront de mieux comprendre comment un langage hormonal commun, qui s'est développé au cours de l'évolution, a contribué à l'interaction entre les plantes et les champignons mycorhiziens depuis qu'elles ont colonisé le milieu terrestre.

International audience; Inflammation is not only the first line of defense of the organism but is also required in many reproductive processes such as ovulation, corpus luteum development, luteolysis, uterine clearance after insemination and post partum. Nevertheless, if excessive or persistent, inflammation can switch from a positive mechanism to a deleterious process, impairing oocyte quality and embryo development. Not only uterine but also non genital inflammatory sites can depreciate reproductive performances, with a carry over effect of 2 to 4 months. Since the metabolic challenges of the peripartum transition period make difficult for the cow to control inflammation, dairy cows are frequently in a pro-inflammatory stage, suggesting that inflammation, rather than infection, is a limiting factor of fertility in modern dairy cows. Within the first week after calving, cows have to mount an intense inflammatory response to the bacterial invasion of the uterine cavity with the challenge of being able to switch it off in no more than 5-6 weeks. The absence of neutrophils on endometrial smear is associated with the highest success rate at insemination. Since a fine tuning - rather than an absence - of inflammation is required along the reproductive cycle, anti-inflammatory drugs do not allow any improvement of pregnancy rate, except in the specific case of embryo transfer. Appropriate management of the transition period (especially nutritional) and in a long term perspective, genetic selection contribute to improve the aptitude of cows to controls the intensity of inflammatory process.

International audience; Rapid adjustment of plant physiology and development to external fluctuations is critical for sessile organism, giving a singular interest to network signaling controlling these mechanisms. Among many adaptation processes, root plasticity is primordial to optimize nutrient acquisition but relies on a complex network integrating local and systemic (root shoot) signaling. Indeed, locally, plants invest resource in soil area where nutrients are available and systemically they adjust nutrient acquisition to the whole plant demand.Our main goal is to decipher systemic signaling underlying the perception of nitrate heterogeneous provision, in Arabidopsis. Using the split-root system, in which physically isolated root systems of the same plant were challenged with different environments, we previously demonstrated that cytokinin biosynthesis constitutes one critical component of root-shoot-root communication. By combining the use of cytokinin mutants with hormone measurements, transcriptomic analysis, nitrate uptake assays, and root growth measurements, we show that root to shoot trans-zeatin (tZ) translocation is likely crucial for long distance signaling controlling rapid sentinel gene regulation and long-term functional acclimation to heterogeneous nitrate supply. Interestingly, shoot transcriptome profiling revealed that glutamate/glutamine metabolism is likely a target of tZ root-to-shoot translocation, prompting an interesting hypothesis regarding shoot-to-root communication. Finally, this study also highlights tZ-independent pathways triggered by variation into nitrogen supply.

The aim of the present work was to test the efficiency of the phytoextraction process involving the use of exogenous phytohormone (cytokinins, CKs) and fertilizer (nitrogen, N) treatments in phytotechnologies to address risk management in “recalcitrant areas”. The CKs and N treatments, alone or combined (CKs + N) in a Modulated Application (MA), were tested on the crop plant Helianthus annuus, common to Mediterranean area, fast growing and with high biomass production. Plants were grown on boron (B) contaminated sediments (collected from a geothermal area located in Tuscany (Italy). Plant growth, B uptake, together with plant stress parameters were investigated. Boron is easily taken up and translocated by some crop plants, but the high phytotoxicity can dramatically impact the plant growth and consequently the applicability and efficiency of the phytoextraction process. As indicators of plant stress, oxidative balance and photosynthetic parameters were investigated to give a deeper insight of phytotoxic mechanisms. Results showed that while each treatment (CKs and N alone) had significantly positive effects on plant health, the MA treatment provided a synergistic effect on morphological parameters and biomass production as a whole. After MA treatment, plants showed antioxidant activity comparable to that of the control (unpolluted sediments) and showed an increase of net photosynthesis. Moreover, our data showed very high values of B uptake and translocation (about 800 mg kg−1 in shoots), without any alteration triggered by the treatments (CKs and N alone or combined in MA). B phytoextraction resulted increased about fivefold with the MA treatments, while each treatment alone increased only two or three folds when treated with either CKs or N. The MA treatment is not “contaminant specific”, so it could be applied in other “recalcitrant areas” where different types of contaminations occur, in order to overcome limitations of plant growth.

Cytokinin signaling is a key regulatory pathway of many aspects in plant development and environmental stresses. Herein, we initiated the identification and functional characterization of the five CHASE-containing histidine kinases (CHK) in the economically important Malus domestica species. These cytokinin receptors named MdCHK2, MdCHK3a/MdCHK3b, and MdCHK4a/MdCHK4b by homology with Arabidopsis AHK clearly displayed three distinct profiles. The three groups exhibited architectural variations, especially in the N-terminal part including the cytokinin sensing domain. Using a yeast complementation assay, we showed that MdCHK2 perceives a broad spectrum of cytokinins with a substantial sensitivity whereas both MdCHK4 homologs exhibit a narrow spectrum. Both MdCHK3 homologs perceived some cytokinins but surprisingly they exhibited a basal constitutive activity. Interaction studies revealed that MdCHK2, MdCHK4a, and MdCHK4b homodimerized whereas MdCHK3a and MdCHK3b did not. Finally, qPCR analysis and bioinformatics approach pointed out contrasted expression patterns among the three MdCHK groups as well as distinct sets of co-expressed genes. Our study characterized for the first time the five cytokinin receptors in apple tree and provided a framework for their further functional studies.

Substantial progress has been made in the understanding of early stages of the symbiotic interaction between legume plants and rhizobium bacteria. Those include the specific recognition of symbiotic partners, the initiation of bacterial infection in root hair cells, and the inception of a specific organ in the root cortex, the nodule. Increasingly complex regulatory networks have been uncovered in which cytokinin (CK) phytohormones play essential roles in different aspects of early symbiotic stages. Intriguingly, these roles can be either positive or negative, cell autonomous or non-cell autonomous, and vary, depending on time, root tissues, and possibly legume species. Recent developments on CK symbiotic functions and interconnections with other signaling pathways during nodule initiation are the focus of this review.

It is well established that plant hormones such as auxins, cytokinins (CKs), and abscisic acid (ABA) not only govern important plant physiological traits but are key players in plant–microbe interactions. A poorly appreciated fact, however, is that both microbes and animals produce and perceive plant hormones and their mimics. Moreover, dietary plant hormones impact on human physiological process such as glucose assimilation, inflammation, and cell division. This leads us to wonder whether plant hormones could ensure functions in microbes per se as well as in animal–microbe interactions. We propose here and explore the hypothesis that plant hormones play roles in animal–microbiota relationships, with consequences for human health.

While the role of many cytokines during infection is revealed, the implication of several recently discovered cytokines during infection is not completely understood. The involvement of currently described interleukin-32 (IL-32) in infectious mastitis (an inflammation of the mammary gland), caused by two prevalent mastitis pathogens, Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) , was not investigated so far. We determined expression of IL-32, IL-6 and IL-8 in S. aureus- as compared to E.coli-infected bovine mammary gland epithelial cells. Using live bacteria we found that in S. aureus-infected cells, induction of IL-6 and IL-8 expression was less pronounced than in E. coli-infected cells. Notably, IL-32 expression was decreased in S. aureus-infected cells, while it was increased in E. coli-infected cells. IL-32 expression in E. coli- and S. aureus-infected udder tissue resembles in vitro responses. We identified the staphylococcal phenol-soluble modulin (PSM) peptides as key contributors to these effects, as IL-32, IL-6 and IL-8 expression by epithelial cells exposed to psm mutant strains was significantly increased as compared to cells exposed to the isogenic S. aureus wild-type strain, indicating that PSMs inhibit the production of these interleukins. The use of genetically complemented strains confirmed this observation. Inasmuch as the decreased expression of IL-32, which is involved in dendritic cell maturation, impairs immune responses, our results support a PSM-dependent mechanism that allows for the development of chronic S aureus-related mastitis.

El desarrollo de técnicas de micropropagación en orquídeas ofrece múltiples ventajas como la reproducción de genotipos superiores, raros, estériles o libres de enfermedades. El objetivo del presente trabajo fue evaluar el uso de suplementos orgánicos (agua de coco y pulpa de banana) en el crecimiento y desarrollo in vitro de plántulas del híbrido intergenérico Laeliocattleya (Hadrolaelia purpurata x Cattleya intermedia), comparando su respuesta con fitorreguladores de auxinas y citoquininas. Para ello se realizaron los siguientes cuatro experimentos: 1. Suplementos orgánicos comparados con diferentes concentraciones de kinetina (N6-furfuril amino purina; KIN) y ácido naftalenacético (ANA), 2. Suplementos orgánicos comparados con diferentes concentraciones de ácido 3-indolbutírico (IBA) y N6-benzyladenine (6BA), 3. Suplementos orgánicos comparados con diferentes concentraciones de IBA y KIN, y 4. Suplementos orgánicos comparados con diferentes concentraciones de ANA y 6BA. El suplemento orgánico de agua de coco fue estadísticamente superior al resto (P

Türkiye, Myrsinaceae familyasına giren Cyclamen cinsi bakımından büyük bir potansiyele sahiptir. Ülkemizde, altısı endemik 10 siklamen türü doğal olarak yetişmektedir. Siklamen çeşitlerinin geliştirilmesinde klasik ıslah yöntemleri yanında günümüzde biyoteknolojik yöntemler de kullanılmaktadır. Ancak, siklamende görülen kendileme depresyonu, farklı ploidi seviyeleri, kendine uyuşmazlık ve abortif embriyo oluşumu nedeniyle klasik yöntemlerle siklamen ıslahı oldukça zordur. Bu nedenle in vitro haploidizasyon, siklamen ıslahında yaygın olarak kullanılmaktadır. Bu çalışmada in vitro haploidizasyon ve in vitro çoğaltımın ilk aşaması olarak bitki doku kültürü ortamlarının ve hormonların etkisi araştırılmıştır. Çalışmada bitkisel materyal olarak, Cyclamen persicum ve Cyclamen pseudibericum'un ovül eksplantları, ortam olarak ise 2,4-dichlorophenoxyaceticacid (2,4-D) ve 6-(γ ,γ –dimethylallylamino) purine' (2iP)'in farklı konsantrasyonlarını içeren ½MS ve B5 ortamları kullanılmıştır. Çalışma sonucunda, Cyclamen persicum türünde en iyi kallus oluşum oranı; 1,5 mg.L-1 (2,4-D ve 0,5 mg.L-1 2iP içeren B5 ortamından (%70 oranında) elde edilmiştir. Cyclamen pseudibericum türü için ise kallus oluşum oranı %48 olmuş ve 1 mg.L-1 2,4-D ve 0,3 mg.L-1 2iP içeren B5 ortamından elde edilmiştir. Cyclamen persicum türünde en yüksek embriyo uyartımı 1,5 mg.L-1 2,4-D ve 0,8 mg.L-1 2iP içeren ½MS ortamında gelişen kalluslardan elde edilmiştir (%55). Ancak Cyclamen pseudibericum türünde besi ortamlarının hiç birinden embriyo uyartımı sağlanamamıştır. Sonuç olarak, kallus uyartımında B5 ortamı, ½MS ortamından daha iyi sonuç vermesine rağmen embriyo uyartımı için yüksek oksin ve düşük sitokinin konsatrasyonuna sahip ½MS ortamı daha iyi bulunmuş ve 2,0 mg.L-1 2,4-D ve 0,5 mg.L-1 2iP içeren ½MS ortamından 42 adet bitkicik elde edilmiştir. Turkey has a great potential in view of Cyclamen genus that belong to Myrsinaceae family. Ten cyclamen species are naturally grown in our country and six of them are endemic to the country. Besides using classical breeding methods to develop new cyclamen cultivars, biotechnological methods have been used currently. However, classical crossing methods in cyclamen breeding are difficult due to selfing depression different ploidy levels, self-incompatibility and abortive embryo formation. Therefore, in vitro haploidisation commonly used in cyclamen breeding. In this study, we investigated effects of different tissue culture media and plant hormones as a first of in vitro haploidisation and propagation. We used ovule explants from Cyclamen persicum and Cyclamen pseudibericum on ½MS and B5 culture mediums supplemented with different concentration of 2,4-dichlorophenoxyaceticacid (2,4-D) and 6-(γ ,γ –dimethylallylamino) purine (2iP).According to results best callus formation rate was 70% obtained from B5 medium supplemented with 1.5 mg.L-1 2,4-D and 0,5 mg.L-1 2iP in Cyclamen persicum. It was 48% on B5 medium supplemented with 1 mg.L-1 2,4-D and 0.3 mg.L-1 2iP in Cyclamen pseudibericum. In Cyclamen persicum, highest embryo induction rate was 55% obtained from callus regenerated on ½MS medium supplemented with 1.5 mg.L-1 2,4-D and 0.8 mg.L-1 2iP. However, embryo induction was not obtained on any medium for Cyclamen pseudibericum. Overall, ½MS culture medium supplemented with high auxin and low cytokinin concentration is better for embryo induction although B5 medium was better than ½MS medium for callus formation and 42 plantlets obtained only from callus regenerated on ½MS medium supplemented with 2.0 mg.L-1 2.4-D and 0,5 mg.L-1 2iP. 71

Plants form the basis of the food webs that sustain animal life. Exogenous factors, such as nutrients and sunlight, and endogenous factors, such as hormones, cooperate to control both the growth and the development of plants. We assessed how Arabidopsis thaliana integrated nutrient and hormone signaling pathways to control root growth and development by investigating the effects of combinatorial treatment with the nutrients nitrate and ammonium; the hormones auxin, cytokinin, and abscisic acid; and all binary combinations of these factors. We monitored and integrated short-term genome-wide changes in gene expression over hours and longterm effects on root development and architecture over several days. Our analysis revealed trends in nutrient and hormonal signal cross-talk and feedback, including responses that exhibited logic gate behavior, which means that they were triggered only when specific combinations of signals were present. From the data, we developed a multivariate network model comprising the signaling molecules, the early gene expression modulation, and the subsequent changes in root phenotypes. This multivariate network model pinpoints several genes that play key roles in the control of root development and may help understand how eukaryotes manage multifactorial signaling inputs.

The impact of infection by the low-virulent ASFV/NH/P68 (NHV) and the highly virulent ASFV/L60 (L60) isolates on porcine macrophages was assessed through the quantification of IFNalpha, TNFalpha, IL12p40, TGFbeta and ASFV genes by real-time PCR at 2, 4 and 6 h post-infection. Increased IFNalpha, TNFalpha and IL12p40 expression was found in infection with NHV, in which expression of TGFbeta was lower than in infection with L60. Principal component analysis showed a positive interaction of cytokines involved in cellular immune mechanisms, namely IFNalpha and IL12p40 in the NHV infection. Quantification by ELISA confirmed higher production of IFNalpha, TNFalpha and IL12p40 in the NHV-infected macrophages. Overall, our studies reinforce and clarify the effect of the NHV infection by targeting cellular and cellular-based immune responses relevant for pig survival against ASFV infection.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:03:25Z No. of bitstreams: 1 S1413-70542007000500001.pdf: 303547 bytes, checksum: d9641f657c00b36f23be0dd30310fd63 (MD5) Made available in DSpace on 2013-08-22T19:03:25Z (GMT). No. of bitstreams: 1 S1413-70542007000500001.pdf: 303547 bytes, checksum: d9641f657c00b36f23be0dd30310fd63 (MD5) Previous issue date: 2007-10-01 Made available in DSpace on 2013-09-30T20:05:17Z (GMT). No. of bitstreams: 2 S1413-70542007000500001.pdf: 303547 bytes, checksum: d9641f657c00b36f23be0dd30310fd63 (MD5) S1413-70542007000500001.pdf.txt: 30189 bytes, checksum: eacb898efae98e73deb574c7d30db8dd (MD5) Previous issue date: 2007-10-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:53:24Z No. of bitstreams: 2 S1413-70542007000500001.pdf: 303547 bytes, checksum: d9641f657c00b36f23be0dd30310fd63 (MD5) S1413-70542007000500001.pdf.txt: 30189 bytes, checksum: eacb898efae98e73deb574c7d30db8dd (MD5) Made available in DSpace on 2014-05-20T13:53:24Z (GMT). No. of bitstreams: 2 S1413-70542007000500001.pdf: 303547 bytes, checksum: d9641f657c00b36f23be0dd30310fd63 (MD5) S1413-70542007000500001.pdf.txt: 30189 bytes, checksum: eacb898efae98e73deb574c7d30db8dd (MD5) Previous issue date: 2007-10-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação para o Desenvolvimento da UNESP (FUNDUNESP) A Cattleya é considerada a mais bela e procurada de todas as orquídeas. Esta demanda cria a necessidade do desenvolvimento de técnicas mais eficazes de propagação que ofereçam maior número de indivíduos em período de tempo e espaço físico reduzido, além de manterem a uniformidade e a identidade genotípica para atender às exigências do mercado. Objetivou-se neste trabalho avaliar a regeneração de meristemas provenientes de gemas cultivadas in vitro do híbrido Blc Owen Holmes Ponkan x Brassavola digbiana nº 2, em diferentes concentrações de BAP (6-benzilaminopurina). Gemas com 37 dias de cultivo foram inoculadas em meio de cultura líquido com reguladores vegetais, nos respectivos tratamentos: T1 - MS líquido + água de coco; T2 - MS + água de coco + 0,1 mg L-1 ANA (ácido naftaleno acético) + 0,2 mg L-1 BAP; T3 - MS + água de coco + 0,1 mg L-1 ANA + 0,5 mg L-1 BAP; T4 - MS + água de coco + 0,1 mg L-1 ANA + 1,0 mg L-1 BAP. Os subcultivos se realizaram aos 18, 32, 62, 93, 123, 153 e 184 dias e as avaliações foram feitas observando-se a porcentagem de sobrevivência, contaminação, oxidação e regeneração dos explantes. Observou-se que 86,7% dos explantes sobreviveram e aos 93 dias ocorreu o primeiro sinal de regeneração. BAP (1,0 mg L-1) promoveu resultados satisfatórios com a formação de 132 perfilhos aos 184 dias de cultivo, potencializando a técnica para produção em escala comercial dessa espécie. Cattleya is considered the most beautiful and searched among all orchids. This fact creates the need of developing more effective propagation techniques to become available a greater number of individuals in reduced periods of time and physical space, besides maintenance of uniformity and genotipic identity to meet the needs of the market. The objective of this work was to evaluate the regeneration of the cultivated shoot meristematic in vitro of the hybrid Blc Owen Holmes Ponkan x Brassavola digbiana nº 2 in the different BAP (6-benzylaminopurine) concentrations. The explants were inoculated in the liquid medium with or without growth regulators in the following combinations: T1: MS liquid + coconut water; T2: MS + coconut water + 0.1 mg L-1 NAA + 0.2 mg L-1 BAP; T3: MS + coconut water + 0.1 mg L-1 NAA + 0.5 mg L-1 BAP and T4: MS + coconut water + 0.1 mg L-1 NAA + 1.0 mg L-1 BAP. Subcultivatings and evaluations (percentage of survival, contamination, medium color, oxidation and regeneration) were made at 18, 32, 62, 93, 123, 153, and 184 days. At 93 days the first signs of regeneration were observed. BAP (1.0 mg L-1) enhanced good results with the formation of 132 sprouts in the 184 days, concluded that the utilization of BAP improved morphogenic response with a higher number of shoots. Universidade Estadual Paulista Júlio de Mesquita Filho Departamento de Química e Bioquímica Universidade Estadual Paulista Júlio de Mesquita Filho Departamento de Química e Bioquímica

Key message Hypocotyl semi-dwarfism in BP was only due to less cell number. Daylight was required to inhibit cell division in the mutant. ABA, IAA and cytokinins would be involved. Abstract Coffea arabica 'Laurina' is a natural mutant of Coffea arabica 'Bourbon' (B) and is known under the trade name 'Bourbon Pointu' (BP). Under daylight, the laurina mutation leads to pleiotropic effects, including semi-dwarf hypocotyls. At the opposite, semi-dwarfism of BP seedlings disappeared under darkness conditions. The first step was to describe the morphological impact of the mutation in seedlings growing under daylight by comparison with seedlings growing under darkness. As the hypocotyl length was mainly affected, the second step was to investigate histological modifications in the organ comparing B and BP seedlings growing under daylight. Result of this investigation indicated that the mutation does not impact on cell length. Moreover, cytometry analyses showed absence of endoreduplication. Actually, the mutation influenced the cell number and this effect appeared before the 40th day after sowing. The length difference of hypocotyls between B and BP was due to lower cell number in BP, indicating possible involvement of phytohormones. Investigations showed the decrease of cytokinin and auxin levels in BP compared to B, while the cytokinin/auxin ratio remained constant in both varieties. By contrast, abscisic acid content increased in BP. Concurrently these results indicate the lowering of cell division, due to the mutation.

Background Today, when more than 60% of animal diseases are zoonotic, understanding their origin and development and identifying protective immune responses in ruminants are major challenges. Robust, efficient and cost-effective tools are preconditions to solve these challenges. Cytokines play a key role in the main mechanisms by which the immune system is balanced in response to infectious pathogens. The cytokine balance has thus become the focus of research to characterize immune response in ruminants. Currently, SYBR Green reverse transcriptase quantitative PCR (RT-qPCR) is the most widely method used to investigate cytokine gene expression in ruminants, but the conditions in which the many assays are carried out vary considerably and need to be properly evaluated. Accordingly, the quantification of gene expression by RT-qPCR requires normalization by multiple reference genes. The objective of the present study was thus to develop an RT-qPCR assay to simultaneously quantify the expression of several cytokines and reference genes in three ruminant species. In this paper, we detail each stage of the experimental protocol, check validation parameters and report assay performances, following MIQE guidelines. Results Ten novel primer sets were designed to quantify five cytokine genes (IL-4, IL-10, IL-12B, IFN-γ and TNF-α) and five reference genes (ACTB, GAPDH, H3F3A, PPIA and YWHAZ) in cattle, sheep, and goats. All the primer sets were designed to span exon-exon boundaries and use the same hybridization temperature. Each stage of the RT-qPCR method was detailed; their specificity and efficiency checked, proved and are reported here, demonstrating the reproducibility of our method, which is capable of detecting low levels of cytokine mRNA up to one copy whatever the species. Finally, we checked the stability of candidate reference gene expression, performed absolute quantification of cytokine and reference gene mRNA in whole blood samples and relative expression of cytokine mRNA in stimulated PBMC samples. Conclusions We have developed a novel RT-qPCR assay for the simultaneous relative quantification of five major cytokines in cattle, sheep and goats, and their accurate normalization by five reference genes. This accurate and easily reproducible tool can be used to investigate ruminant immune responses and is widely accessible to the veterinary research community. Electronic supplementary material The online version of this article (doi:10.1186/s12917-015-0382-0) contains supplementary material, which is available to authorized users.

İşyeri ortamında sensitizan kimyasallara maruziyet solunum yoluyla ilgili çok ciddi mesleki hastalıklara yol açmaktadır. Son yıllarda gıdalarda katkı maddesi olarak kullanılan Diasetil maddesinin bronşiyolitis obliterans ve astım gibi hastalıklarla ilişkili olduğu ve bu nedenle üretim yerinde çalışan işçi sağlığı için tehlike oluşturduğu düşünülmektedir. Tez çalışmamızda bu tehlikenin boyutunu değerlendirmek üzere Diasetil'in sensitizan karakteristiğini ortaya koymayı amaçladık. Negatif, pozitif ve çözücü kontrol gruplarıyla birlikte Diasetil'in sensitizasyon potansiyelinin belirlenmesi için hayvan refahının gözetildiği ve radyoaktif olmayan ex vivo Lokal Lenf Düğümü Testi (LLNA):BrdU-ELISA yöntemi kısa süreli ve kronik maruziyet protokolleri ile yürütülmüştür. İmmün reaksiyonlarda sitokinlerin ve antikorların başlıca rolünün olmasından ve bu zamana kadar Diasetil'in tüm sitokin profili aydınlatılmadığından, sitokin profilleme ve fare IgE testi çalışmaya dahil edilmiştir. Deneylerimiz sonucunda Diasetil'in %10 (ağırlık/hacim) konsantrasyonda çözücü kontrole kıyasla lenfosit poliferasyonunu anlamlı şekilde indüklemediği, aynı konsantrasyonda her iki maruziyet süresinde de anlamlı IgE antikoru artışına neden olduğu gözlenmiştir. Bu çalışmanın Diasetil'in işçi sağlığına yönelik taşıdığı riskin değerlendirilmesine ve sağlık otoritelerince gerekli düzenlemelerin yapılmasına katkı sağlayacağı düşünülmektedir. Exposure to chemical sensitizers in workplace settings leads to significant occupational diseases related to respiratory tract. In recent years diacetyl, which is used as a food additive, is considered to be associated with various diseases such as bronchiolitis obliterans and asthma and therefore diacetyl is thought to be as an important hazard for worker's health. With this thesis study we aimed to characterize the sensitization potency of diacetyl for the purpose of its hazard evaluation. For determining the sensitization potency of diacetyl with negative, pozitive and vehicle control groups, animal welfare caring and non-radioactive ex vivo Local Lymph Node Assay (LLNA):BrdU-ELISA method with short term and chronic exposure protocols was performed. Due to significant role of cytokines and antibodies in immune reactions and with limited knowledge about the whole cytokine profile of diacetyl up to now, cytokine profiling and Mouse IgE test were incorporated into research. As a consequence of our experiments, lymphocyte proliferation induced with a concentration of 10% (w/v) of diacetyl was not found as statistically significant in comparison with vehicle control. The same concentration of the test material induced statistically significant IgE antibody levels in comparison with vehicle control group in both exposure protocols. In conclusion this research supports the studies in the field of risk assessment of diacetyl for occupational safety and contibutes to health authorities to adopt new regulations 131

Phyllotaxis, the spatio-temporal pattern of organogenesis at the shoot apical meristem, emerges in large part from inhibitory fields consisting in auxin-depleted areas centered on organs. We recently demonstrated the existence of an additional hormone-based inhibitory field generated by Arabidopsis Histidine Phosphotransfer Protein 6 (AHP6), an inhibitor of cytokinin signaling. We have shown that the spatio-temporal distribution of AHP6 in the meristem is essential for optimizing the rhythmicity of organ initiation. Here, we further analyzed AHP6 expression using fluorescent whole mount mRNA in situ hybridization and demonstrate a precise control of AHP6 level and expression domain over time. While we previously showed a regulation of AHP6 directly downstream of auxin, we show here that AHP6 transcription is unlikely influenced by cytokinin distribution in the meristem. Finally, we provide evidence that cytokinins and auxin might act synergistically during organ initiation, providing a plausible explanation for how AHP6 regulates phyllotaxis.

Giriş-Amaç: Kronik böbrek hastalarında kardiyovasküler hastalıklar mortalitenin en önemli nedenidir. Beslenme bozukluğunun ileri formu olan protein-enerji tükenme sendromu bu hızlanmış mortaliteye katkıda bulunmaktadır. Kronik böbrek hastalarında iştahsızlık, iştahı düzenleyen hormonlar arasındaki dengenin bozulması, inflamasyon, diyaliz işlemi, istirahat halindeki enerji harcanmasının artması, insülin direnci ve D-vitamini eksikliği gibi hormonal bozukluklar, metabolik asidoz, anabolizmanın yavaşlaması, fiziksel aktivitenin azalması ve komorbid hastalıklar protein-enerji tükenme sendromunun gelişmesine neden olmaktadır. Çalışmamızdaki amaç; iştahı arttırma özelliği olan agouti-related peptit düzeyinin, kronik böbrek yetmezliği bulunan protein-enerji tükenme sendromlu hastalardaki klinik durum, antropometrik ölçümler ve inflamatuvar sitokinler ile arasındaki ilişkiyi incelemektir.Gereç-Yöntem: Çalışmaya 76 hasta alındı. Protein-enerji tükenme sendromu kritelerinin 3 veya daha fazlasına sahip olan 38 hasta protein-enerji tükenme sendromu grubunu, 2 kritere sahip olan 27 hasta malnutrisyon grubunu, 11 sağlıklı birey kontrol grubunu oluşturdu. Genel biyokimyasal parametreler, tam kan sayımı, IL-1, 6, TNF-α, agouti-related peptit tahlilleri çalışıldı. Vücut kitle indeksi, günlük protein alımı, orta kol kas çevresi alanı hesaplandı. Hastaların kullandıkları ilaçlar ile eşlik eden hastalıklar kaydedildi.Bulgular: Çalışmaya aldığımız hastaların %51'inde protein-enerji tükenme sendromu mevcuttu.Anlamlı olarak agouti-related peptit düzeyi protein-enerji tükenme sendromu grubunda kontrol grubuna göre daha düşük, TNF-α ve CRP düzeyi ise daha yüksekti. Üç grup içinde albumin değeri ve vücut kitle indeksi en düşük protein enerji tükenme sendromu grubunda tespit edildi. Orta kol kas çevresi alanı ve günlük protein alımı, kontrol grubuna göre yine protein-enerji tükenme sendromu grubunda anlamlı daha düşüktü. Agouti-related peptit düzeyi ile günlük protein alımı arasında pozitif yönlü, orta kol kas çevresi alanı ile CRP ve albumin ile TNF-α arasıda negatif yönlü bir korelasyon tespit edildi. Protein-enerji tükenme sendromu grubundaki hastaların %97'sinde yağsız doku kitle indeksinde azalma mevcuttu. Hemodiyaliz ve periton diyalizi yapan protein-enerji tükenme sendromlu hastalar arasında agouti-related peptit düzeyi açısından anlamlı farklılık yoktu. Tartışma: Kronik böbrek hastalarında protein-enerji tükenme sendromu yaygın bulunmaktadır. Günlük protein alımının azalması ve artmış inflamasyon, protein-enerji tükenme sendromu gelişimine katkıda bulunabilir. Protein-enerji tükenme sendromlu hastalarda agouti-related peptit düzeyi ciddi düzeyde düşüktür ve bu durum günlük protein alımını etkiliyebilir. Agouti-related peptit düzeyini arttırmaya yönelik girişimler, protein-enerji tükenme sendromu gelişme riskini azaltabilir. Introduction, aim: Cardiovascular diseases are the most important causes of mortality in patients with chronic kidney disease. Protein energy wasting syndrome, an advanced form of malnutrition, has contributed to this accelerated mortality. Anorexia, the imbalance of hormones which regulate appetite, inflammation, dialysis, the incresed resting energy expenditure, hormonal derangement such as insulin resistance and vitamin D deficiency, metabolic acidosis, decreased anabolism, decreased physical activity and comorbidities, have caused the development of protein energy wasting syndrome in patients with chronic kidney disease. The aim of our study is to evaluate the correalation between the level of agouti-related peptide which increase apetite and clinic, anthropometric measurements and cytokines in patients with chronic renal failure and protein energy wasting syndrome.Material-methods: Seventy-six patients were included in this study. Thirty-eight patients who have three or more criteria of protein energy wasting syndrome, 27 patients who have two criteria and 11 healthy individuals created protein energy wasting syndrome group, malnutrition group, and control group respectively. The general biochemical parameters, complete blood count, IL-1, 6, TNF-α, agouti-related peptide were measured. Body mass index, daily protein intake, mid-arm muscle circumference area were calculated. Comorbidities and received drugs were enrolled.Results: Fifty-one percent of our patients had protein energy wasting syndrome criteria. Agouti-related peptide was significantly lower and TNF-α, CRP levels were significantly higher in protein energy wasting syndrome group than control group. The lowest albumin value and body mass index were identified in protein energy wasting syndrome group among the three groups. According to control group, mid-arm circumference area and dietary protein intake were significantly lower in protein energy wasting syndrome group. The positive correlation between agouti-related peptide and daily protein intake, the negative correlation between mid-arm circumference area and CRP, between albumin and TNF-α were found. There was a reduction in lean tissue index in 97% of patients in protein energy wasting syndrome group. There was no significant difference according to agouti-related peptide level between hemodialysis and peritoneal dialysis patients with protein energy wasting syndrome.Discussion: Protein energy wasting syndrome are common in patients with chronic kidney disease. The reduction of daily protein intake and the increased inflammation may contribute to the development of protein energy wasting syndrome. Agouti-related peptide level is seriously low in patients with protein energy wasting syndrome. This situation may influence daily protein intake. The approaches which increase agouti-related peptide level may reduce the risk of development of protein energy wasting syndrome. 78

BACKGROUND: Selecting experimental material at the optimal physiological stage is of paramount importance for successful cryopreservation. OBJECTIVE: The study was to investigate the effect of the physiological state of grapevine buds on their regrowth after liquid nitrogen exposure. METHODS: In a first set of experiments, we tested the regrowth of cryopreserved buds sampled from microcuttings cultured on shooting medium containing benzylaminopurine or zeatin riboside for various durations. In a second set of experiments, we studied the regrowth after liquid nitrogen exposure of buds sampled from different positions on the stem of in vitro plantlets. RESULTS: Regrowth of cryopreserved buds sampled from microcuttings was higher (30%), compared to buds sampled directly from in vitro plantlets (23%), for all culture durations of microcuttings on shooting medium tested (2-6 weeks). Addition of cytokinin in the shooting medium improved regrowth of cryopreserved buds compared to buds sampled from microcuttings cultured on medium devoid of growth regulators; however similar results were obtained with the two cytokinins tested. Buds sampled on nodes 3-4 and 6-7 (from the top of the stem) displayed higher regrowth compared to shoot tips. No significant differences were noted in regrowth after cryopreservation between buds sampled from microcuttings produced from the terminal node, or nodes 3-4 and 6-7. CONCLUSION: The physiological state of the plant material is important for cryopreservation success. Actively growing buds sampled from microcuttings displayed higher regrowth compared to buds sampled directly on in vitro plantlets.

Bu çalışmada, streptozotosin ile diyabet oluşturulmuş sıçanlarda Saccharomyces cerevisiae kaynaklı ticari beta glukanın antioksidan ve immunoprotektif etkileri çeşitli parametreler kullanılarak araştırılmıştır. Oksidatif stres ve doku hasarının belirlenmesi için, karaciğer ve böbrek dokularında malondialdehid (MDA) ve miyeloperoksidaz (MPO) aktiviteleri, antioksidan etki için süperoksid dismutaz (SOD), katalaz (CAT), glutatyon peroksidaz (GPx) ve glutatyon redüktaz (GR) enzimlerinin aktiviteleri, immunoprotektif etkiler için inflamatuvar (TNF-α ve IL-6) ve antiinflamatuvar (IL-10) sitokinlerin serum düzeyleri incelendi. Çalışmada 40 adet Spraque Dawley erkek sıçan, 4 gruba ayrıldı. Grup 1 kontrol (n=10), grup 2 diyabet (n=7), grup 3 beta glukan (n=10) ve grup 4 (n=7) diyabet+beta glukan olarak belirlendi. Deneysel diyabet, grup 2 ve 4'teki sıçanlara intraperitoneal olarak 60 mg/kg STZ enjeksiyonu ile oluşturuldu. Enjeksiyondan 24 saat sonra grup 2 ve grup 4'teki sıçanlara beta glukan (50 mg/kg) 10 gün süreyle nazogastrik sonda ile oral olarak verildi. Deney başlangıcı ve sonunda vücut ağırlıkları ölçülmüş, diyabet ve diyabet+beta glukan gruplarındaki hayvanların deney sonu vücut ağırlıklarında anlamlı bir azalma görüldü (p< 0.001). Diyabet ve diyabet+beta glukan gruplarının glukoz düzeyleri deney başındaki glukoz düzeyi ile karşılaştırıldığında anlamlı bir artış tespit edilmiştir (p< 0.001). Beta glukan grubunda ise anlamlı bir değişiklik görülmemiştir. Açlık kan şekeri ise, diyabet grubunda yükselmiş, diyabet+beta glukan grubunda ise diyabet grubu ile kıyaslandığında anlamlı bir azalma tespit edilmiştir (p

In this study we observed the way in which cytokinine influence the growing of corn seedlings and the conditions in which they present a nitrogen deficiency. We worked with two different concentrations of cytokinine: 1% and respectively 0.1%, introduced into 800 ml of a complete Knop solution, a Knop solution containing a double quantity of nitrogen, compared to the control solution, a Knop solution containing half of the quantity of nitrogen and a Knop solution absent in nitrogen. Daily measurings was made for 14 days, considering the length of the roots and shots, in every experimental variant. This study shows that a direct proportionality exists between the growing rate of the corn seedlings and the nitrogen concentration. It was also showed that the average of the growing rate for a period of 14 days was of 0.28 cm at a nitrogen concentration on 2.0 g/l, and only 0.05 cm in case of nitrogen deficiency. Analyzing the average of the shots growing rate (cm), it was 0.39 in case of the variant with cytokinine 0.1%, 0.45 in case of the variant with cytokinine 1% and 0.36 in case of the control sample.

Balıkesir Üniversitesi, Fen Bilimleri Enstitüsü, Biyoloji Ana Bilim Dalı, ADAMTS'ler (A Disintegrin And Metalloprotease domain with ThromboSpondin type I motifs) embriyonik gelişim, anjiyogenez ve kıkırdak degredasyonu gibi önemli fizyolojik süreçlerde görev yapan metaloproteazlardır. Bu ailenin üyelerinden ADAMTS2 fibriler prokollajenlerin amino uçlarının kesilmesinde görev yapmaktadır. ADAMTS2 genindeki mutasyonlar insanlarda Ehler Danlos sendromu tip VIIC'ye neden olmaktadır. Organizma için daha birçok önemli fonksiyonu bulunan ADAMTS2'nin transkripsiyonel regülasyonunun aydınlatılması oldukça önemlidir. Bu çalışmada transkripsiyon başlangıç bölgesinden 760bç'ye kadar uzanan ADAMTS2 promotor bölgesi ve transkripsiyonel olarak en aktif bölgenin belirlenmesi amacıyla üç farklı uzunlukta [-530/+112], [-324/+112], [-180/+112] promotor parçası lusiferaz vektörüne klonlandı. Saos (insan osteosarkoma) hücrelerinde yapılan geçici transfeksiyon analizleri sonucunda bütün parçaların yüksek promotor aktivitesi gösterdiği ve transkripsiyonel aktivite için en küçük promotor parçasının yeterli olduğu belirlendi. Biyoinformatik analizlerle ADAMTS2 promotorunun 110bç'lik 5'UTR bölgesine sahip olduğu, TATA kutusu içermediği, GCkutuları, GA kutuları, CpG adaları, SP1 ve USF için bağlanma motifleri içerdiği belirlendi. Yapılan ko-transfeksiyon çalışmalarıyla USF ve SP1'in ADAMTS2 promotor aktivitesini arttırdığı belirlendi. USF'nin fonksiyonel olarak promotora bağlandığı da EMSA çalışmalarıyla belirlendi. ADAMTS2'nin çalışılan hücre hatlarında en fazla MG-63'te ve en az HUVEC'te ifade olduğu belirlendi. Saos hücrelerinde ADAMTS2 proteini immunfloresans ile işaretlendi. Ayrıca Saos hücrelerinde IL-6 ve IL-1a stimülasyonuyla ADAMTS2 ve ADAMTS3 ekspresyonunun mRNA ve protein düzeyinde arttığı qRT-PCR ve Western blot çalışmalarıyla belirlendi. Benzer MG-63 modelinde de sonuçlar doğrulandı. IL-6'nın ADAMTS2 promotor aktivitesini arttırdığı belirlendi. Saos hücrelerinde yapılan inhibisyon çalışmalarıyla IL-6'nın STAT yolunu ve de-novo protein sentezini aktive ettiği belirlendi. IL-6 sitokininin Saos hücrelerinde hücre proliferasyonuna neden olmazken IL-1a'nın hücre proliferasyonuna neden olduğu MTT çalışmalarıyla belirlendi., ADAMTS (A Disintegrin And Metalloprotease domain with ThromboSpondin type I motifs) are metalloproteinases with demonstrated functionality in embryonic development, angiogenesis and cartilage degradation. One of the members of this family, ADAMTS2 responsible for removal of the amino terminus of the fibrillar procollagens. Mutations in the ADAMTS2 gene causes Ehler Danlos syndrome type VIIC in humans. It is quite important to elucidate the transcriptional regulation of the ADAMTS2 which has many important functions for organism. In this study, ADAMTS2 promoter region extending up to 760bp from the transcription start site and three different portions of the promoter [-530/+112], [-324/+112], [-180/+112] fragments were cloned into luciferase vector to determine transcriptionally most active region. As a result high promoter activity was determined with all constructs in transiently transfected Saos (human osteosarcoma) cells and the smallest construct was enough to exhibit the transcriptional activity. Bioinformatic analyses revealed that TATA-less ADAMTS2 promoter has a 110bp 5'UTR region including GC, GA boxes, CpG islands, SP1 and USF binding motifs. The co-transfection studies determined increased activity of the ADAMTS2 promoter with USF and SP1. Functional binding of USF to ADAMTS2 promoter was determined by EMSA studies. Expression of ADAMTS2 was determined at most MG-63 and at least HUVEC among the studied cell lines. ADAMTS2 protein was labelled in Saos cells by immunofluorescence. Increased ADAMTS2 and ADAMTS3 expression at mRNA and protein levels with IL-6 and IL-1' stimulation of Saos cells was also determined qRT-PCR and Western blotting studies. Results were also confirmed similar MG-63 model. IL-6 was increased activity of the ADAMTS2 promoter. Inhibition studies showed STAT pathway and de-novo protein synthesis activation after IL-6 stimulation in Saos cells. Cell proliferation wasn't determined with IL-6 stimulation in Saos cells by MTT experiments. On the contrary IL-1a caused cell proliferation.

Richard Tafner Zsfassung in dt. und engl. Sprache Graz, Univ., Masterarb., 2013

Plants produce cytokinin (CK) hormones for controlling key developmental processes like source/sink distribution, cell division or programmed cell-death. Some plant pathogens have been shown to produce CKs but the function of this mimicry production by non-tumor inducing pathogens, has yet to be established. Here we identify a gene required for CK biosynthesis, CKS1, in the rice blast fungus Magnaporthe oryzae. The fungal-secreted CKs are likely perceived by the plant during infection since the transcriptional regulation of rice CK-responsive genes is altered in plants infected by the mutants in which CKS1 gene was deleted. Although cks1 mutants showed normal in vitro growth and development, they were severely affected for in planta growth and virulence. Moreover, we showed that the cks1 mutant triggered enhanced induction of plant defenses as manifested by an elevated oxidative burst and expression of defense-related markers. In addition, the contents of sugars and key amino acids for fungal growth were altered in and around the infection site by the cks1 mutant in a different manner than by the control strain. These results suggest that fungal-derived CKs are key effectors required for dampening host defenses and affecting sugar and amino acid distribution in and around the infection site., Author Summary The role of plant-like hormonal compounds produced by fungal pathogens during infection has not been elucidated. Here we identified a conserved gene in most fungi, required for cytokinin production by the rice blast fungus and for its full virulence. Fungal-derived cytokinins are likely potent inhibitors of plant immunity. They are also needed to maintain elevated sugar contents at the site of infection and to drain or consume essential amino acids at, and around, the infection site. Thus, cytokinins represent the second example, after the bacterially-produced coronatine, of plant hormones hijacked by pathogens to successfully invade plant tissues. These findings also suggest that this invasion strategy could be widely conserved among fungi.

Mercury-contaminated soils from a petrochemical plant in southern Italy were investigated to assess the phytoextraction efficiency of crop plants treated with the phytohormone, cytokinine (CK foliar treatment), and with the thioligand, ammonium thiosulfate (TS, soil application). Plant biomass, evapotranspiration, Hg uptake and distribution in plant tissues following treatment were compared. Results indicate the effectiveness of CK in increasing plant biomass and the evapotranspiration rate while IS treatment promoted soil Hg solubility and availability. The simultaneous addition of CK and IS treatments increased Hg uptake and translocation in both tested plants with up to 248 and 232% in Brassica juncea (Indian mustard) and Helianthus annuus (sunflower) respectively. B. juncea was more effective in Hg uptake, whereas H. annuus gave better response regarding plant biomass production. The effectiveness of the treatments was confirmed by the calculation of Hg phytoextraction and evaluation of labile-Hg residue in the soil after plant growth. In one growing cycle the plants subject to simultaneous CK and IS treatment significantly reduced labile-Hg pools that were characterized by the soil sequential extraction, but did not significantly affect the pseudototal metal content in the soil. Results support the use of plant growth regulators in the assisted phytoextraction process for Hg-contaminated soils. (C) 2012 Elsevier B.V. All rights reserved.

Amaç. BH tekrarlayan oral ve genital ülserler, üveit ve deri lezyonları ile karakterize kronik inflamatuar bir hastalıktır. BH etiyopatogenezi henüz ayrıntılı olarak tanımlanamamakla birlikte, genetik, enfeksiyon, immun kompleksler, otoantikorlar ve oksidatif stres Behçet nedenleri arasında ileri sürülmektedir. Çalışmamızda inflamatuar reaksiyonlarda rol oynadığı düşünülen IL-1ß, IL-2, IL-6, TNF-?, TLR4 ve Hsp60 genlerinin ekspresyon düzeylerine bakılarak hastalığın etyolojisindeki otoenflamatuar olayların açıklığa kavuşturulması ve hastalığın tedavisine moleküler genetik temelli katkıda bulunulması amaçlandı.Materyal ve Metot. Çalışmamızda, Behçet tanısı almış 40 hasta ve kontrol grubu olarak sağlıklı 30 bireyde RT-PZR yöntemi ile IL-1ß, IL-2, IL-6, TNF-? TLR4 ve Hsp60 genlerinin ekspresyon düzeyleri çalışıldı. İstatistiksel verilerin değerlendirilmesinde SPSS versiyon 13, Ki-kare ve One Way Anova istatistiksel analiz yöntemleri kullanıldı.Bulgular. Behçet hastalarında ekspresyon seviyelerine bakılan 6 genden 3'ünün (IL-1ß, IL-6 ve TLR4) ekspresyonlarında istatistiksel olarak kontrol grubuna göre önemli derecede azalma gözlendi. Hastaların kullandıkları siklosporin ve kolşisin gibi ilaçların ekspresyon üzerine etkisi, ilaçları kullanan gruplar arasında karşılaştırıldı. Elde edilen bulgulara göre IL-1ß, IL-6 ve TLR4 genlerinin ekspresyon seviyeleri arasında bu ilaçların etkisi istatistiksel olarak anlamlı bulunmadı.Sonuç. Sonuç olarak, bu çalışmada Behçet hastalarının kullandıkları immünsüpresörlerin IL-1ß, IL-6 ve TLR4 genlerinin ekspresyon seviyelerini azaltıcı yönde etki gösterdiği belirlendi.Anahtar kelimeler: Behçet Hastalığı, Ekspresyon, Sitokin Aim. BD is a chronic inflamatory condition characteriged with recurrent oral and genital ulcers, uveit and skin lessions. Although the pathogenesis of the disease remains are not clearly understood, genetics, infections, immune coplex, autoantibodies, and oxidative stres are counted among possible reasons. In our study, by examining expression lewels of IL-1ß, IL-2, IL-6 TNF-?, TLR4 and Hsp60 genes with the aim of molecular genetically based contribution to the treatment of the disease and with the aim of revealing autoinflammature events in the disease etiology.Material and Methods. In our study, we determined the expression lewels of IL-1ß, IL-2, IL-6 TNF-?, TLR4 and Hsp60 genes by means of RT-PCR in 40 patiens with Behçet?s diagnoses and the control group consisting of 30 healthy individuals. SPSS version 13, Chi-square and One Way Anova statistical analysis methods were used for statistical evaluation of data.Results. It was observed that there was significant reduction in 3 of (IL-1ß, IL-6, TLR4) 6 genes expressions in Behçet?s patients by means of statistical analysis. Effects of drugs, such as cyclosporine and colchicine that used by patients, were compared to both drugs user groups. According to the findings obtained, the distinctions between IL-1ß, IL-6, TLR4 gene expressions was not statistically significant in both drug users.Conclusion. In this study, we determined that, immunosuppressors that used by Behcet?s patients had impact in reducing way on expresssion lewels of IL-1ß, IL-6 and TLR4 genes.Key Words; Behçet?s Disease, Cytokine, Expression 79

Roots of young Norway spruce seedlings were incubated under hydroculture conditions in a synthetic nutrient medium containing either 3H-isopentenyladenosine, isopentenyladenosine or zeatin riboside. When feeding with 3H-isopentenyladenosine a new radiaolabelled metabolite was found in the feeding solution as well as in root extracts. Isopentenyladenosine and zeatin riboside were metabolised and for both compounds an unknown metabolite was detected in the feeding solution. The metabolites were purified by solid phase extraction, HPLC and partially characterised. A major characteristic of the metabolites is their reactivity in the presence of NH4OH, which results in the formation of the cytokinin bases isopentenyladenine or zeatin, respectively. UV-spectra and the chemical characteristics indicate that the new metabolites are closely related. The GC-MS analysis revealed, that the metabolites are true derivatives of isopentenyladenine and zeatin. The biogenesis of the new metabolites is discussed with regard to plant microbial interactions.

The agrobacterial isopentenyltransferase (ipt) gene, under the control of its native promoter, was transferred to poplar (Populus tremula x P. alba) by an Agrobacterium co-cultivation method. The ipt-transformed stem explants developed calli that regenerated many buds in the absence of exogenous cytokinins. Microcuttings of the ipt transformants exhibited frequently branching shoots with short internodes that were unable to root. In this material, the concentrations of zeatin, zeatin riboside and isopentenyladenosine, determined by an indirect enzyme-linked immunosorbent assay (ELISA), were 4.8-. 17.1- and 14.6-fold higher, respectively, than in non-transformed shoots. Results are discussed with regard to cytokinin metabolism.

Promising immunomodulatory properties of select strains of dairy[i] propionibacteria[/i]. 4 .Congress of European Microbiologists FEMS 2011

Effect of GA3 and paclobutrazol on adventitious shoot regeneration of two [i]Pelargonium[/i] sp.. 7. ISHS on In Vitro Culture & Horticultural Breeding

Alzheimer Hastalığı (AH), ileri yaştaki demansların en sık formudur. Kognitif fonksiyonlarda ilerleyici bozulma ile karakterizedir. Klinik pratikte, AH'nın kesin kriterlerine rağmen tanısı sekonder nedenlerin ve diğer demansif hastalıkların dışlanması ile konur. AH'da erken tanı ve hastalığın erken evrelerinde müdahale, tedavinin etkinliği açısından büyük önem taşır. Bu sebeple sağlıklı bireylerde olası bir yatkınlığın tespiti hastalığa karşı alınabilecek önlemler bakımından yararlı olacaktır. Araştırmamızda adiposit kökenli bir sitokin olan Visfatin'in -948 G-T polimorfizminin AH'na yatkınlıktaki etkisi incelenmiştir.Çalışmaya 40 Alzheimer hastası ve 40 sağlıklı kontrol alınmıştır. AH olguları Dokuz Eylül Üniversitesi Tıp Fakültesi Nöroloji Anabilim Dalı Demans polikliniğinde takip edilen hastalardan ardışık sırayla 20.06.2009-20.03.2010 tarihleri arasında alınmıştır. Kontrol grubu, hastalar ile benzer demografik özelliklere sahip gönüllü sağlıklı kişilerden oluşturulmuştur. Hasta ve kontrollere Mini Mental Durum Testi (MMDT), Sözel Bellek Süreçleri Testi (SBST), Benton Yüz Tanıma Testi, Kategorik Akıcılık Testi uygulanmıştır. AH ve kontrol olgularının kan örneklerinden Visfatin -948 G-T polimorfizmi real-time PCR ile allel spesifik primerler kullanılarak erime eğrisi analizi ile incelenmiştir. Visfatin -948 G-T polimorfizmi dışında serum açlık kan şekeri (AKŞ), Low Density Lipid (LDL), High Density Lipid (HDL), trigliserid, total kolesterol ve C-Reaktif Protein (CRP) düzeyleri de ölçülmüştür.AH ve kontrol grubu -948 G-T polimorfizmi açısından değerlendirildiğinde iki grup arasında istatistiksel olarak anlamlı bir fark bulunmamıştır.Çalışmamızdaki örneklemde Visfatin -948 G-T polimorfizminin AH'na yatkınlığa neden olmadığı saptanmıştır. Ayrıca klinik bulgularla -948 G-T polimorfizmi arasında da bir ilişki saptanamamıştır. AH patogenezinde Visfatin'in olası rolünü araştıracak daha geniş örneklem grubunda yapılacak yeni çalışmalara ihtiyaç vardır. Alzheimer?s disease (AD) is the most common form of dementia in elderly. It is characterized by progressive deterioration of cognitive functions. In clinical practice, current criteria for diagnosis of AD are still largely based on the exclusion of secondary causes and other dementive disorders. Eary diagnosis and treatment at early stages are very important at AD for the efficacy of treatment. Pointing out a possible predisposition will be helpful for taking precautions against AD. In our study, we examined the effect of Visfatin -948 G-T polymorphism as a predisposition for AD.In the present study, 40 AD patients and 40 healthy control subjects were included. AD patients were selected consecutively from Dementia outpatient clinic in Dokuz Eylül University, Department of Neurology between the dates of 20.06.2009-20.03.2010. Control cases were selected from the healthy elderly volunteer persons with similar demographic features. We studied -948 G-T polymorphism from AD patients and healty controls peripheric blood samples with real-time PCR using allel specific primer and melting curve analysis. Beside -948 G-T polymorphism we investigated serum fasting glucose levels (FGL) , LDL, HDL, triglyceride, total cholesterol and C-Reactive Protein (CRP). All the patients and controls also were examined with the Mini Mental State Examination (MMSE), The Rey Auditory Verbal Learning Test (RAVLT), Benton Facial Recognition Test and Category Naming Test.There were no significant difference between patients and healthy cases for -948 G-T polymorphism.According to our experiment group, we show that the -948 G-T polymorphism does not cause predisposition for AD. Future studies with larger numbers of patients and controls are necessary to clarify exact role of Visfatin in AD pathogenesis. 55