Analysis of Microsatellite Polymorphism in Inbred Knockout Mice

Previously, we found that the genotype of 42 out of 198 mouse microsatellite loci, which are distributed among all chromosomes except the Y chromosome, changed from monomorphism to polymorphism (CMP) in a genetically modified inbred mouse strain. In this study, we further examined whether CMP also relates to the homologous recombination in gene knockout (KO) mouse strains. The same 42 microsatellite loci were analyzed by polymerase chain reaction (PCR) in 29 KO inbred mouse strains via short tandem sequence repeat (STR) scanning and direct sequence cloning to justify microsatellite polymorphisms. The C57BL/6J and 129 mouse strains, from which these 29 KO mice were derived, were chosen as the background controls. The results indicated that 10 out of 42 (23.8%) loci showed CMP in some of these mouse strains. Except for the trinucleotide repeat locus of D3Mit22, which had microsatellite CMP in strain number 9, the core sequences of the remaining 41 loci were dinucleotide repeats, and 9 out of 41 (21.95%) showed CMPs among detected mouse strains. However, 11 out of 29 (37.9%) KO mice strains were recognized as having CMPs. The popular dinucleotide motifs in CMP were (TG)n (50%, 2/4), followed by (GT)n (27.27%, 3/11) and (CA)n (23.08%, 3/13). The microsatellite CMP in (CT)n and (AG)n repeats were 20% (1/5). According to cloning sequencing results, 6 KO mouse strains showed insertions of nucleotides whereas 1 showed a deletion. Furthermore, 2 loci (D13Mit3 and D14Mit102) revealed CMP in 2 strains, and mouse strain number 9 showed CMPs in two loci (D3Mit22 and D13Mit3) simultaneously. Collectively, these results indicated that microsatellite polymorphisms were present in the examined inbred KO mice

Daidzein Prevents the Increase in CD4+CD28null T Cells and B Lymphopoesis in Ovariectomized Mice: A Key Mechanism for Anti-Osteoclastogenic Effect

Estrogen deficiency leads to an upregulation of TNF-α producing T cells and B-lymphopoesis which augments osteoclastogenesis. Estrogen deficiency also increases the population of premature senescent CD4+CD28null T cells which secrete a higher amount of TNF-α thus leading to enhanced osteoclastogenesis. Isoflavonoids like daidzein and genistein are found mostly in soybeans, legumes, and peas. These share structural similarity with 17β-stradiol (E2) and have osteoprotective role. This study explores the effect of daidzein (Daid) on the proliferation of TNF-α producing T cells, premature senescent T cells and B cell lymphopoesis under estrogen deficient conditions. For this study adult Balb/c mice were treated with Daid at 10 mg/kg body weight dose by oral gavage daily post ovariectomy (Ovx). After six weeks animals were autopsied and bone marrow and spleen cells were collected for FACS analysis. Blood serum was collected for ELISA. It was observed that Ovx mice treated with Daid for six weeks show reduction in Ovx induced expansion of CD4+ T cells in bone marrow and spleen when analysed by flow cytometry. Estrogen deficiency led to increased prevalence of TNF-α secreting CD4+CD28null T cells, however, treatment with Daid increased the percentage of CD4+CD28+ T cells. Co-culture of CD4+CD28null T cells and bone marrow resulted in enhanced osteoclastogenesis as evident by increased tartarate resistant acid phosphatase (TRAP) expression, an osteoclast marker. However, treatment with Daid resulted in reduced osteoclastogenesis in CD4+CD28null T cells and bone marrow cell co-culture. Daid also regulated B lymphopoesis and decreased mRNA levels of RANKL in B220+ cells. Taken together, we propose that one of the mechanisms by which Daid prevents bone loss is by reversing the detrimental immune changes as a result of estrogen deficiency

The Regulation of MS-KIF18A Expression and Cross Talk with Estrogen Receptor

This study provides a novel view on the interactions between the MS-KIF18A, a kinesin protein, and estrogen receptor alpha (ERα) which were studied in vivo and in vitro. Additionally, the regulation of MS-KIF18A expression by estrogen was investigated at the gene and protein levels. An association between recombinant proteins; ERα and MS-KIF18A was demonstrated in vitro in a pull down assay. Such interactions were proven also for endogenous proteins in MBA-15 cells were detected prominently in the cytoplasm and are up-regulated by estrogen. Additionally, an association between these proteins and the transcription factor NF-κB was identified. MS-KIF18A mRNA expression was measured in vivo in relation to age and estrogen level in mice and rats models. A decrease in MS-KIF18A mRNA level was measured in old and in OVX-estrogen depleted rats as compared to young animals. The low MS-KIF18A mRNA expression in OVX rats was restored by estrogen treatment. We studied the regulation of MS-KIF18A transcription by estrogen using the luciferase reporter gene and chromatin immuno-percipitation (ChIP) assays. The luciferase reporter gene assay demonstrated an increase in MS-KIF18A promoter activity in response to 10−8 M estrogen and 10−7M ICI-182,780. Complimentary, the ChIP assay quantified the binding of ERα and pcJun to the MS-KIF18A promoter that was enhanced in cells treated by estrogen and ICI-182,780. In addition, cells treated by estrogen expressed higher levels of MS-KIF18A mRNA and protein and the protein turnover in MBA-15 cells was accelerated. Presented data demonstrated that ERα is a defined cargo of MS-KIF18A and added novel insight on the role of estrogen in regulation of MS-KIF18A expression both in vivo and in vitro

Exercise and bone health across the lifespan

With ageing, bone tissue undergoes significant compositional, architectural and metabolic alterations potentially leading to osteoporosis. Osteoporosis is the most prevalent bone disorder, which is characterised by progressive bone weakening and an increased risk of fragility fractures. Although this metabolic disease is conventionally associated with ageing and menopause, the predisposing factors are thought to be established during childhood and adolescence. In light of this, exercise interventions implemented during maturation are likely to be highly beneficial as part of a long-term strategy to maximise peak bone mass and hence delay the onset of age- or menopause-related osteoporosis. This notion is supported by data on exercise interventions implemented during childhood and adolescence, which confirmed that weight-bearing activity, particularly if undertaken during peripubertal development, is capable of generating a significant osteogenic response leading to bone anabolism. Recent work on human ageing and epigenetics suggests that undertaking exercise after the fourth decade of life is still important, given the anti-ageing effect and health benefits provided, potentially occurring via a delay in telomere shortening and modification of DNA methylation patterns associated with ageing. Exercise is among the primary modifiable factors capable of influencing bone health by preserving bone mass and strength, preventing the death of bone cells and anti-ageing action provided

Virologie / Bakteriologie / Mykologie

141 - Effizienz von Kaliumhypochlorit zur Inaktivierung ausgewählter pilzlicher, bakterieller und viraler PflanzenkrankheitserregerEfficancy of Potassium Hypochlorite (KClO) to inactivate selected plant pathogenic fungi, bacteria and virusesMarlon-Hans Rodríguez, Martina Bandte, Gerhard Fischer, Carmen Büttner142 - Eignung von elektrolytisch generiertem Kaliumhypochlorit zur Inaktivierung von Pflanzenviren in rezirkulierender Nährlösungen im Gewächshausanbau von TomatenAbility of electrolysed produced Potassium Hypochlorite (KClO) to inactivate plant viruses in recirculating nutrient solutions in greenhouse production of tomatosJanine Paulke, Martina Bandte, Carmen Büttner143 - Ultrafiltration und Ultrazentrifugation zur Konzentrierung von Pflanzenviren in NährlösungUltrafiltration and ultracentrifugation as tools to concentrate plant viruses in nutrient solutionJanina Vincenz, Martina Bandte, Carmen Büttner144 - Reinigung doppelsträngiger RNA in Verbindung mit Hochdurchsatzsequenzierung als Werkzeug zum Nachweis von RNA Viren in PflanzenThe combination of double-stranded RNA isolation and deep sequencing as an unspecific diagnostic tool to assess the presence of RNA viruses in plantsTill Lesker, Paul Rentz, Edgar Maiss145 - Impact of silica supplementation on virus infected cucumber culturesRolle der Kieselsäureapplikation Virus infizierter GurkenkulturenSabine Holz, Grzegorz Bartoszewski , Michael Kube, Carmen Büttner146 - Untersuchungen zum Auftreten des Arabis mosaic virus in Birken aus Rovaniemi (Finnland) mit Virus-spezifischen SymptomenInvestigations on the occurence of Arabis mosaic virus in birches from Rovaniemi (Finland) with virus-specific symptomsRichard Pauwels, Markus Rott, Susanne von Bargen, Carmen Büttner147 - Cherry leaf roll virus in Betula spp. in Finland: what do we know about its population diversity?Cherry leaf roll virus in Birken-Arten in Finnland: Was wissen wir über die Populationsdiversität?A. Rumbou, S. von Bargen, M. Rott, R. Jalkanen, C. Büttner148 - Viruserkrankungen im WeinbauViroses in viticultureHenriette Gruber, Patricia Bohnert, Christiane Rieger149 - Molecular analysis of Tobacco rattle virus isolates from potatoes in various parts of GermanyKerstin Lindner, Renate Koenig150 - Detektion und Diversität des European mountain ash ringspot-associated virus (EMARaV) in Ebereschen (Sorbus aucuparia L.) in NorwegenDetection and variability of European mountain ash ringspot-associated virus (EMARaV) in Sorbus aucuparia L. in NorwayTheresa Büttner, Jenny Robel, Hans-Peter Mühlbach, Susanne von Bargen, Carmen Büttner151 - Charakterisierung des European mountain ash ringspot-associated virus (EMARaV) in Mehlbeerenarten (Sorbus spp.)Characterization of the European mountain ash ringspot-associated virus (EMARaV) in whitebeam species (Sorbus spp.)Luisa Dieckmann, Jenny Robel, Susanne von Bargen, Carmen Büttner152 - Vollständige Genomsequenz eines Carrot virus S Isolates aus Meerfenchel aus SpanienW. Menzel, P. Menzel, S. Winter153 - Nachweis und vollständige Sequenzierung eines Carla- und eines Potex-virus aus Epiphyullum spec.Detection and complete sequence of a Carla- and Potexvirus in Epiphyllum spec.Edgar Maiss, Paul Rentz, Annette Hohe, Rosa Herbst154 - Analysis of mixed populations of latent viruses of apple and rubbery wood disease of apple using new generation sequencingAnalyse von Mischpopulationen latenter Apfelviren und der Gummiholzkrankheit an Apfel mittels HochdurchsatzsequenzierungVladimir Jakovljevic, Patricia Otten, Jonathon Blake, Wilhelm Jelkmann155 - Experiments on transmission of viroids under glass and longevity of viroid RNA in detached leaves under different storage conditionsThi Thu Vo, Heinz-Wilhelm Dehne, Stephan Winter, Joachim Hamacher156 - Phytoplasmen in Schleswig-HolsteinPhytoplasmas in the state of Schleswig-HolsteinG. Henkel, C. Willmer, M. Wunderlich, B. Golecki157 . Phytoplasmen verändern das Dufststoffbouquet ihres pflanzlichen LebensraumsPlant volatile emission is affected by phytoplasma infectionMargit Rid, Kai Lukat, Svenja Hoferer, Jürgen Gross159 - Ist das Wurzelbild ein Sortierungsmerkmal für durch Candidatus Phytoplasma pyri verursachten Birnenverfall?Is the root file a sorting feature for Pear decline caused by Canditatus Phytoplama pyri?Georg Henkel, Claudia Willmer, Bernd Kaland, Bettina Golecki160 - Die Bedeutung von β-Caryophyllen als Lockstoff für die Apfeltriebsucht übertragende Blattsaugerart Cacopsylla pictaThe impact of β-caryophyllene as attractant for the Apple Proliferation transmitting insect Cacopsylla pictaConstanze Mesca, Svenja Hoferer, Jürgen Gross161 - Echte Mehltauarten an Beet- und BalkonpflanzenSpecies of powdery mildews on bedding plantsUlrike Brielmaier-Liebetanz162 - Echter Mehltau an Petersilie – Untersuchungen zum WirtspflanzenspektrumPowdery Mildew of Parsley – studies on the host rangePeggy Marx, Ute Gärber163 - Falscher Mehltau an Petersilie – Untersuchungen zum Wirtspflanzenspektrum und molekularbiologische CharakterisierungDowny mildew of parsley – studies on the host range and molecular characterizationGabriele Leinhos, Hermann-Josef Krauthausen, Frank Brändle164 - Welkekrankheit an Euonymus japonicaWilt disease on Euonymus japonicaUlrike Brielmaier-Liebetanz, Roswitha Ulrich, Stefan Wagner, Sabine Werres165 - Taxonomische Analyse der mikrobiellen Gemeinschaft von Zuckerrüben unter unterschiedlichen Lagerbedingungen mittels Hochdurchsatz-Amplikonsequenzierung von unterschiedlichen MarkergenenTaxonomic analysis of the microbial community in stored sugar beets using highthroughput sequencing of different marker genesSebastian Liebe, Daniel Wibberg, Anika Winkler, Alfred Pühler, Andreas Schlüter, Mark Varrelmann166 - Molecular characterization of a novel mycovirus found in Rhizoctonia solani AG 2-2IIIBMolekulare Charakterisierung eines neuen Mycovirus aus Rhizoctonia solani AG 2-2 IIIBAnika Bartholomäus, Mark Varrelman

Interrelationships between Yeast Ribosomal Protein Assembly Events and Transient Ribosome Biogenesis Factors Interactions in Early Pre-Ribosomes

Early steps of eukaryotic ribosome biogenesis require a large set of ribosome biogenesis factors which transiently interact with nascent rRNA precursors (pre-rRNA). Most likely, concomitant with that initial contacts between ribosomal proteins (r-proteins) and ribosome precursors (pre-ribosomes) are established which are converted into robust interactions between pre-rRNA and r-proteins during the course of ribosome maturation. Here we analysed the interrelationship between r-protein assembly events and the transient interactions of ribosome biogenesis factors with early pre-ribosomal intermediates termed 90S pre-ribosomes or small ribosomal subunit (SSU) processome in yeast cells. We observed that components of the SSU processome UTP-A and UTP-B sub-modules were recruited to early pre-ribosomes independently of all tested r-proteins. On the other hand, groups of SSU processome components were identified whose association with early pre-ribosomes was affected by specific r-protein assembly events in the head-platform interface of the SSU. One of these components, Noc4p, appeared to be itself required for robust incorporation of r-proteins into the SSU head domain. Altogether, the data reveal an emerging network of specific interrelationships between local r-protein assembly events and the functional interactions of SSU processome components with early pre-ribosomes. They point towards some of these components being transient primary pre-rRNA in vivo binders and towards a role for others in coordinating the assembly of major SSU domains