Shaping our future: animal health in a global trading environment

Ireland is currently experiencing both the positive and negative effects of global trade in agricultural products. With increasing global competition, there is little doubt that Irish agricultural product must increasingly compete on the basis of quality, rather than price

Acidithiobacillus ferrooxidans metabolism: from genome sequence to industrial applications

<p>Abstract</p> <p>Background</p> <p><it>Acidithiobacillus ferrooxidans </it>is a major participant in consortia of microorganisms used for the industrial recovery of copper (bioleaching or biomining). It is a chemolithoautrophic, γ-proteobacterium using energy from the oxidation of iron- and sulfur-containing minerals for growth. It thrives at extremely low pH (pH 1–2) and fixes both carbon and nitrogen from the atmosphere. It solubilizes copper and other metals from rocks and plays an important role in nutrient and metal biogeochemical cycling in acid environments. The lack of a well-developed system for genetic manipulation has prevented thorough exploration of its physiology. Also, confusion has been caused by prior metabolic models constructed based upon the examination of multiple, and sometimes distantly related, strains of the microorganism.</p> <p>Results</p> <p>The genome of the type strain <it>A. ferrooxidans </it>ATCC 23270 was sequenced and annotated to identify general features and provide a framework for <it>in silico </it>metabolic reconstruction. Earlier models of iron and sulfur oxidation, biofilm formation, quorum sensing, inorganic ion uptake, and amino acid metabolism are confirmed and extended. Initial models are presented for central carbon metabolism, anaerobic metabolism (including sulfur reduction, hydrogen metabolism and nitrogen fixation), stress responses, DNA repair, and metal and toxic compound fluxes.</p> <p>Conclusion</p> <p>Bioinformatics analysis provides a valuable platform for gene discovery and functional prediction that helps explain the activity of <it>A. ferrooxidans </it>in industrial bioleaching and its role as a primary producer in acidic environments. An analysis of the genome of the type strain provides a coherent view of its gene content and metabolic potential.</p

Lactic Acidosis in a Newborn With Adrenal Calcifications

A patient is reported who presented in the newborn period with an unusual combination of congenital lactic acidosis and bilateral calcifications in the adrenal medulla, visible on standard abdominal x-ray and ultrasound examination. At birth, the proband was hypotonic and dystrophic. She developed respiratory insufficiency, cardiomegaly, and hepatomegaly and died at the age of 38 d. Examination of postmortem heart muscle revealed multiple areas of myocardial infarction with dystrophic calcifications. In the medulla of the adrenal glands, foci of necrosis and calcifications, and in the liver, multiple zones of necrosis and iron deposition were detected. Biochemical-analysis in heart muscle revealed a decreased activity of complex IV of the oxidative phosphorylation (OXPHOS) and in liver a combined deficiency involving the complexes I, III, IV, and V. The findings were suggestive of a defect in biosynthesis of the mitochondrially encoded subunits of the OXPHOS complexes. Extensive analysis of the proband's mitochondrial DNA revealed neither pathogenic deletions and point mutations nor copy number alterations. Relative amounts of mitochondrial transcripts for the ribosomal mitochondrial 12S rRNA (12S) and mitochondrial 16S rRNA (16S) were significantly increased suggesting a compensatory mechanism involving the transcription machinery to low levels of translation. The underlying molecular defect has not been identified yet

Engineering of human-induced pluripotent stem cells for precise disease modeling

Stem cell technologies and gene editing techniques are two of the most promising recent developments in biomedicine. The ability to reprogram common human cells into induced pluripotent stem cells (hiPSCs) and turn them into the cells of interest has already become a powerful research tool, thus providing a unique platform for disease studies. In combination with the use of designer nucleases approach to repair or to introduce disease-causing mutations, both are valuable in developing personalized disease models. This chapter provides an overview on designer nucleases-based gene editing in hiPSCs, describing the principles of CRISPR/Cas systems along with consecutive methodological steps such as nucleases selection, isolation, and genotyping of modified hiPSC clones with emphasis on the crucial role of isogenic cell lines in disease modeling. Moreover, the production of rare or complex genotypes in patient cell lines requires efficient and streamlined gene editing technologies. However, precise genome editing applications rely on infrequent homology-directed repair (HDR), with the abundant nonhomologous end joining (NHEJ) formed indels presenting a barrier to achieving high rates of precise sequence modification. The methods presented here are supported by theoretical framework to allow for the incorporation of inevitable improvements to achieve either higher rates of gene editing by promotion of HDR over NHEJ or application of different CRISPR/Cas platforms for robust and multiplex gene editing, toward decoding of neurodevelopmental as well as for modeling of late onset disorders by fast-forwarding the biological clock. Due to easy in theory but laborious and inefficient in practice, the precise and efficient genome editing in hiPSCs could be only achieved by the proper combination of the described methods in the process. This eventually would lead to generation of wide range of disease models for decoding of sporadic, polygenic, undiagnosed, and rare disorders using the adequate experimental design following appropriate gene editing toolbox selection