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by Jason G. Walling, Leslie A. Zalapa, Marcus A. Vinje Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a popular method for measuring transcript abundance. The most commonly used method of interpretation is relative quantification and thus necessitates the use of normalization controls (i.e. reference genes) to standardize transcript abundance. The most popular gene targets for RT-qPCR are housekeeping genes because they are thought to maintain a static transcript level among a variety of samples. However, more recent studies have shown, several housekeeping genes are not reliably stable. This is the first study to examine the potential of several reference genes for use in RT-qPCR normalization during barley malting. The process of malting barley mechanizes the imbibition and subsequent germination of barley seeds under controlled conditions. Malt quality is controlled by many pleiotropic genes that are determined by examining the result of physiological cha ...

by Andrea Kosovac, Jes Johannesen, Oliver Krstić, Milana Mitrović, Tatjana Cvrković, Ivo Toševski, Jelena Jović The stolbur phytoplasma vector Hyalesthes obsoletus is generally considered as a polyphagous species associated with numerous wild and cultivated plants. However, recent research in southeastern Europe, the distribution centre of H . obsoletus and the area of most stolbur-inflicted crop diseases, points toward specific host-plant associations of the vector, indicating specific vector-based transmission routes. Here, we study the specificity of populations associated with four host-plants using mitochondrial and nuclear genetic markers, and we evaluate the evolution of host-shifts in H . obsoletus . Host-plant use was confirmed for Convolvulus arvensis , Urtica dioica , Vitex agnus-castus and Crepis foetida . Mitochondrial genetic analysis showed sympatric occurrence of three phylogenetic lineages that were ecologically delineated by host-plant preference, but were morph ...

by Philipp Solbach, Patrick Chhatwal, Sabrina Woltemate, Evelina Tacconelli, Michael Buhl, Markus Gerhard, Christoph K. Thoeringer, Maria J. G. T. Vehreschild, Nathalie Jazmati, Jan Rupp, Michael P. Manns, Oliver Bachmann, Sebastian Suerbaum Background Clostridium difficile infection (CDI) is a major cause of hospital-acquired diarrhea. Secondary bile acids were shown to confer resistance to colonization by C . difficile . 7α-dehydroxylation is a key step in transformation of primary to secondary bile acids and required genes have been located in a single bile acid-inducible (bai) operon in C . scindens as well as in C . hiranonis , two Clostridium sp. recently reported to protect against C . difficile colonization. Aim To analyze baiCD gene abundance in C . difficile positive and negative fecal samples. Material & methods A species-specific qPCR for detecting baiCD genes was established. Fecal samples of patients with CDI, asymptomatic toxigenic C . difficile colonization (TCD), non ...

by Cassio Geremia Freire, Admir José Giachini, João Peterson Pereira Gardin, Ana Claudia Rodrigues, Renato Luis Vieira, César Milton Baratto, Simone Silmara Werner, Bernardo Haas Abreu Like many other species of trees native to the Brazilian Mata Atlântica (Atlantic Forest), the Myrtaceae, such as the Red Araza ( Psidium cattleianum Sabine), are widely cited as arbuscular mycorrhizal formers. Nevertheless, recent studies show evidence that Myrtaceae from different tropical, subtropical and neotropical ecosystems can also prompt the formation of ectomycorrhizae, indicating that this species' ectomycorrhizal status should be further explored. Because of this, this research effort studied the in vitro interaction between the Red Araza and two ectomycorrhizal fungi isolates, belonging to the Pisolithus microcarpus (D17) and Scleroderma citrinum (UFSC-Sc133) species. An analysis was performed to determine the formation of ectomycorrhizal structures, or lack thereof, and the development ...

by Lisiery N. Paiatto, Fernanda G. D. Silva, Áureo T. Yamada, Wirla M. S. C. Tamashiro, Patricia U. Simioni Introduction In addition to conventional therapies, several new strategies have been proposed for modulating autoimmune diseases, including the adoptive transfer of immunological cells. In this context, dendritic cells (DCs) appear to be one of the most promising treatments for autoimmune disorders. The present study aimed to evaluate the effects of adoptive transfer of DCs obtained from both naïve and ovalbumin (OVA)-tolerant mice on the severity of TNBS induced colitis and analyze the eventual protective mechanisms. Methods and results To induce oral tolerance, BALB/c mice were fed 4mg/mL OVA solution for seven consecutive days. Spleen DCs were isolated from tolerant (tDC) and naïve (nDC) mice, and then adoptively transferred to syngeneic mice. Three days later, colitis was induced in DC treated mice by intrarectal instillation of 100μg2,4,6-trinitrobenzenesulfonic acid (T ...

by Chung Hyun Cho, Ji Won Choi, Daryl W. Lam, Kyeong Mi Kim, Hwan Su Yoon The red algal subclass Nemaliophycidae includes both marine and freshwater taxa that contribute to more than half of the freshwater species in Rhodophyta. Given that these taxa inhabit diverse habitats, the Nemaliophycidae is a suitable model for studying environmental adaptation. For this purpose, we characterized plastid genomes of two freshwater species, Kumanoa americana (Batrachospermales) and Thorea hispida (Thoreales), and one marine species Palmaria palmata (Palmariales). Comparative genome analysis identified seven genes ( ycf 34, ycf 35, ycf 37, ycf 46, ycf 91, grx , and pbs A) that were different among marine and freshwater species. Among currently available red algal plastid genomes (127), four genes ( pbs A, ycf 34, ycf 35, ycf 37) were retained in most of the marine species. Among these, the pbs A gene, known for encoding heme oxygenase, had two additional copies ( HMOX1 and HMOX2 ) that were newly ...

by Dorien Clarisse, Karlien Van Wesemael, Jan Tavernier, Fritz Offner, Ilse M. Beck, Karolien De Bosscher Glucocorticoids (GCs) are a cornerstone in the treatment of lymphoid malignancies such as multiple myeloma (MM) and acute lymphoblastic leukemia (ALL). Yet, prolonged GC use is hampered by deleterious GC-related side effects and the emergence of GC resistance. To tackle and overcome these GC-related problems, the applicability of selective glucocorticoid receptor agonists and modulators was studied, in search of fewer side-effects and at least equal therapeutic efficacy as classic GCs. Compound A (CpdA) is a prototypical example of such a selective glucocorticoid receptor modulator and does not support GR-mediated transactivation. Here, we examined whether the combination of CpdA with the classic GC dexamethasone (Dex) may improve GC responsiveness of MM and ALL cell lines. We find that the combination of Dex and CpdA does not substantially enhance GC-mediated cell killing. In lin ...

by Sohyun Cho, Lari M. Hiott, John B. Barrett, Elizabeth A. McMillan, Sandra L. House, Shaheen B. Humayoun, Eric S. Adams, Charlene R. Jackson, Jonathan G. Frye Surface waters are important sources of water for drinking, industrial, agricultural, and recreational uses; hence, contamination of water by fecal, pathogenic, or antimicrobial resistant (AR) bacteria is a major environmental and public health concern. However, very little data is available on prevalence of these bacteria in surface water throughout a watershed. This study aimed to characterize Escherichia coli present in the Upper Oconee Watershed, a mixed-use watershed in Athens, GA, USA for potential pathogenicity and AR. E . coli were enumerated by colony counts, cultured by enrichment and direct plating, and characterized by phylo-groups, diarrheagenic pathotypes, and antimicrobial susceptibility. From the analysis, 99.3% (455/458) of the total samples were positive for E . coli resulting in 496 isolates. E . coli counts ...

by Allison R. Sirois, Daniela A. Deny, Samantha R. Baierl, Katia S. George, Sarah J. Moore Mesothelin is a cell surface protein that is overexpressed in numerous cancers, including breast, ovarian, lung, liver, and pancreatic tumors. Aberrant expression of mesothelin has been shown to promote tumor progression and metastasis through interaction with established tumor biomarker CA125. Therefore, molecules that specifically bind to mesothelin have potential therapeutic and diagnostic applications. However, no mesothelin-targeting molecules are currently approved for routine clinical use. While antibodies that target mesothelin are in development, some clinical applications may require a targeting molecule with an alternative protein fold. For example, non-antibody proteins are more suitable for molecular imaging and may facilitate diverse chemical conjugation strategies to create drug delivery complexes. In this work, we engineered variants of the fibronectin type III domain (Fn3) non-a ...

by Kazumi Matsubara, Yuki Iwasaki, Issei Nishiki, Kazuharu Nomura, Atushi Fujiwara Japanese eel ( Anguilla japonica ) constitutes one of the most important food fish in Japan; accordingly, genome sequencing and linkage mapping have been conducted for the purpose of artificial cultivation. In the next stage, integration of genomic sequences within linkage groups (LG) is required to construct higher-resolution genetic markers for quantitative trait loci mapping and selective breeding of beneficial traits in farming. In order to identify LG1-linked scaffolds from the draft genome assembly (323,776 scaffolds) reported previously, we attempted to isolate chromosomes corresponding to LG1 by flow sorting and subsequent analyses. Initially, single chromosomes were randomly collected by chromosome sorting and subjected to whole-genome amplification (WGA). A total of 60 WGA samples were screened by PCR with primers for a known LG1-linked scaffold, and five positive WGA samples were sequenced by ...