Cytopathological Outcomes of Knocking down Expression of Mitochondrial Complex II Subunits in Dictyostelium discoideum

Mitochondrial Complex II is composed of four core subunits and mutations to any of the subunits result in lowered Complex II activity. Surprisingly, although mutations in any of the subunits can yield similar clinical outcomes, there are distinct differences in the patterns of clinical disease most commonly associated with mutations in different subunits. Thus, mutations to the SdhA subunit most often result in mitochondrial disease phenotypes, whilst mutations to the other subunits SdhB-D more commonly result in tumour formation. The reason the clinical outcomes are so different is unknown. Here, we individually antisense-inhibited three of the Complex II subunits, SdhA, SdhB or SdhC, in the simple model organism Dictyostelium discoideum. Whilst SdhB and SdhC knockdown resulted in growth defects on bacterial lawns, antisense inhibition of SdhA expression resulted in a different pattern of phenotypic defects, including impairments of growth in liquid medium, enhanced intracellular proliferation of the bacterial pathogen Legionella pneumophila and phagocytosis. Knockdown of the individual subunits also produced different abnormalities in mitochondrial function with only SdhA knockdown resulting in broad mitochondrial dysfunction. Furthermore, these defects were shown to be mediated by the chronic activation of the cellular energy sensor AMP-activated protein kinase. Our results are in agreement with a role for loss of function of SdhA but not the other Complex II subunits in impairing mitochondrial oxidative phosphorylation and they suggest a role for AMP-activated protein kinase in mediating the cytopathological outcomes.     

Supplementation with SCFAs Re-Establishes Microbiota Composition and Attenuates Hyperalgesia and Pain in a Mouse Model of NTG-Induced Migraine

Migraine is a common brain-disorder that affects 15% of the population. Converging evidence shows that migraine is associated with gastrointestinal disorders. However, the mechanisms underlying the interaction between the gut and brain in patients with migraine are not clear. In this study, we evaluated the role of the short-chain fatty acids (SCFAs) as sodium propionate (SP) and sodium butyrate (SB) on microbiota profile and intestinal permeability in a mouse model of migraine induced by nitroglycerine (NTG). The mice were orally administered SB and SP at the dose of 10, 30 and 100 mg/kg, 5 min after NTG intraperitoneal injections. Behavioral tests were used to evaluate migraine-like pain. Histological and molecular analyses were performed on the intestine. The composition of the intestinal microbiota was extracted from frozen fecal samples and sequenced with an Illumina MiSeq System. Our results demonstrated that the SP and SB treatments attenuated hyperalgesia and pain following NTG injection. Moreover, SP and SB reduced histological damage in the intestine and restored intestinal permeability and the intestinal microbiota profile. These results provide corroborating evidence that SB and SP exert a protective effect on central sensitization induced by NTG through a modulation of intestinal microbiota, suggesting the potential application of SCFAs as novel supportive therapies for intestinal disfunction associated with migraine.     

The Use of Stories in User Experience Design

Stories capture the characteristics of the design space and audience that designers and engineers need to understand to build a complete and useful software experience. A story is a design communication tool that transcends the cultural divides of multidisciplinary teams and intertwines a technology with its user's goals. This article describes how stories are powerful tools in software design, defines the elements that make a compelling story, and presents the use of stories at IBM from the authors' experience. It also explores the benefits at each phase of the design process and how stories evolve throughout the design process.     

Digital neighborhoods

With the advent of ‘big data’ there is an increased interest in using social media to describe city dynamics. This paper employs geo-located social media data to identify ‘digital neighborhoods’ – those areas in the city where social media is used more often. Starting with geo-located Twitter and Foursquare data for the New York City region in 2014, we applied spatial clustering techniques to detect significant groupings or ‘neighborhoods’ where social media use is high or low. The results show that beyond the business districts, digital neighborhoods occur in communities undergoing shifting socio-demographics. Neighborhoods that are not digitally oriented tend to have higher proportion of minorities and lower incomes, highlighting a social–economic divide in how social media is used in the city. Understanding the differences in these neighborhoods can help city planners interested in generating economic development proposals, civic engagement strategies, and urban design ideas that target these areas.     

Experimental realization of ‘universal homodyne tomography’ with a single local oscillator

We report preliminary experimental measurement of the twin-beam quantum state via optical homodyne tomography using a single local oscillator. The experiment is a realization of the recently reported ‘universal homodyne tomography’ technique. The results agree well with theoretical predictions and reveal the non-classical photon-number correlation between the signal and idler photons of the twin-beam state.     

An NMR-based metabolomic assessment of cultured cobia health in response to dietary manipulation

Commercial aquaculture feeds rely heavily on fishmeal and fish oil, which can be expensive and ecologically unsustainable. To evaluate the efficacy of reduced fishmeal diets for outgrowth, a dietary study was conducted on the finfish cobia, Rachycentron canadum. NMR-based metabolomic techniques were used to assess the effect of decreasing dietary fishmeal on the health of the cobia. Filtered serum ¹H NMR spectra analysed by principal components analysis (PCA) showed cobia fed reduced fishmeal diets were metabolically different than cobia on control diets. In particular, tyrosine and betaine increased in cobia fed reduced fishmeal diets while glucose decreased, suggesting that these cobia were not receiving the necessary nutritional components required for energy and growth. The formulated control diet contributed to enriched growth and significantly elevated lactate levels suggesting enhanced gut microflora metabolism in response to dietary components. The results show that NMR-based metabolomic analysis is a useful tool in aquaculture studies.     

Molecular Mechanisms Underlying TDP-43 Pathology in Cellular and Animal Models of ALS and FTLD

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are neurodegenerative disorders that exist on a disease spectrum due to pathological, clinical and genetic overlap. In up to 97% of ALS cases and ~50% of FTLD cases, the primary pathological protein observed in affected tissues is TDP-43, which is hyperphosphorylated, ubiquitinated and cleaved. The TDP-43 is observed in aggregates that are abnormally located in the cytoplasm. The pathogenicity of TDP-43 cytoplasmic aggregates may be linked with both a loss of nuclear function and a gain of toxic functions. The cellular processes involved in ALS and FTLD disease pathogenesis include changes to RNA splicing, abnormal stress granules, mitochondrial dysfunction, impairments to axonal transport and autophagy, abnormal neuromuscular junctions, endoplasmic reticulum stress and the subsequent induction of the unfolded protein response. Here, we review and discuss the evidence for alterations to these processes that have been reported in cellular and animal models of TDP-43 proteinopathy.     

Repurposing the Pummerer Rearrangement: Determination of Methionine Sulfoxides in Peptides

The reversible oxidation of methionine residues in proteins has emerged as a biologically important post-translational modification. However, detection and quantitation of methionine sulfoxide in proteins is difficult. Our aim is to develop a method for specifically derivatizing methionine sulfoxide residues. We report a Pummerer rearrangement of methionine sulfoxide treated sequentially with trimethylsilyl chloride and then 2-mercaptoimidazole or pyridine-2-thiol to produce a dithioacetal product. This derivative is stable to standard mass spectrometry conditions, and its formation identified oxidized methionine residues. The scope and requirements of dithioacetal formation are reported for methionine sulfoxide and model substrates. The reaction intermediates have been investigated by computational techniques and by ¹³C NMR spectroscopy. These provide evidence for an α-chlorinated intermediate. The derivatization allows for detection and quantitation of methionine sulfoxide in proteins by mass spectrometry and potentially by immunochemical methods.