Insular amyloidosis and diabetes mellitus in a crab-eating macaque (Macaca fascicularis)

Clinicopathologic examination of a crab-eating macaque (Macaca fascicularis) with chronic weight loss and bilateral cataracts revealed high fasting serum glucose, glucosuria and hypercholesterolemia. Clinical signs were eliminated by treatment once a day with isophane insulin suspension. Extensive insular amyloidosis was found microscopically sixty days later.     

Caprylic acid‐induced impurity precipitation from protein A capture column elution pool to enable a two‐chromatography‐step process for monoclonal antibody purification

This article presents the use of caprylic acid (CA) to precipitate impurities from the protein A capture column elution pool for the purification of monoclonal antibodies (mAbs) with the objective of developing a two chromatography step antibody purification process. A CA‐induced impurity precipitation in the protein A column elution pool was evaluated as an alternative method to polishing chromatography techniques for use in the purification of mAbs. Parameters including pH, CA concentrations, mixing time, mAb concentrations, buffer systems, and incubation temperatures were evaluated on their impacts on the impurity removal, high‐molecular weight (HMW) formation and precipitation step yield. Both pH and CA concentration, but not mAb concentrations and buffer systems, are key parameters that can affect host–cell proteins (HCPs) clearance, HMW species, and yield. CA precipitation removes HCPs and some HMW species to the acceptable levels under the optimal conditions. The CA precipitation process is robust at 15–25°C. For all five mAbs tested in this study, the optimal CA concentration range is 0.5–1.0%, while the pH range is from 5.0 to 6.0. A purification process using two chromatography steps (protein A capture column and ion exchange polishing column) in combination with CA‐based impurity precipitation step can be used as a robust downstream process for mAb molecules with a broad range of isoelectric points. Residual CA can be effectively removed by the subsequent polishing cation exchange chromatography. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1515–1525, 2015     

Impact of Altitude on Power Output during Cycling Stage Racing

Purpose

The purpose of this study was to quantify the effects of moderate-high altitude on power output, cadence, speed and heart rate during a multi-day cycling tour.

Methods

Power output, heart rate, speed and cadence were collected from elite male road cyclists during maximal efforts of 5, 15, 30, 60, 240 and 600 s. The efforts were completed in a laboratory power-profile assessment, and spontaneously during a cycling race simulation near sea-level and an international cycling race at moderate-high altitude. Matched data from the laboratory power-profile and the highest maximal mean power output (MMP) and corresponding speed and heart rate recorded during the cycling race simulation and cycling race at moderate-high altitude were compared using paired t-tests. Additionally, all MMP and corresponding speeds and heart rates were binned per 1000m (<1000m, 1000–2000, 2000–3000 and >3000m) according to the average altitude of each ride. Mixed linear modelling was used to compare cycling performance data from each altitude bin.

Results

Power output was similar between the laboratory power-profile and the race simulation, however MMPs for 5–600 s and 15, 60, 240 and 600 s were lower (p ≤ 0.005) during the race at altitude compared with the laboratory power-profile and race simulation, respectively. Furthermore, peak power output and all MMPs were lower (≥ 11.7%, p ≤ 0.001) while racing >3000 m compared with rides completed near sea-level. However, speed associated with MMP 60 and 240 s was greater (p < 0.001) during racing at moderate-high altitude compared with the race simulation near sea-level.

Conclusion

A reduction in oxygen availability as altitude increases leads to attenuation of cycling power output during competition. Decrement in cycling power output at altitude does not seem to affect speed which tended to be greater at higher altitudes.     

Macroecology of Australian Tall Eucalypt Forests: Baseline Data from a Continental-Scale Permanent Plot Network

Tracking the response of forest ecosystems to climate change demands large (≥1 ha) monitoring plots that are repeatedly measured over long time frames and arranged across macro-ecological gradients. Continental scale networks of permanent forest plots have identified links between climate and carbon fluxes by monitoring trends in tree growth, mortality and recruitment. The relationship between tree growth and climate in Australia has been recently articulated through analysis of data from smaller forest plots, but conclusions were limited by (a) absence of data on recruitment and mortality, (b) exclusion of non-eucalypt species, and (c) lack of knowledge of stand age or disturbance histories. To remedy these gaps we established the Ausplots Forest Monitoring Network: a continental scale network of 48 1 ha permanent plots in highly productive tall eucalypt forests in the mature growth stage. These plots are distributed across cool temperate, Mediterranean, subtropical and tropical climates (mean annual precipitation 850 to 1900 mm per year; mean annual temperature 6 to 21°C). Aboveground carbon stocks (AGC) in these forests are dominated by eucalypts (90% of AGC) whilst non-eucalypts in the understorey dominated species diversity and tree abundance (84% of species; 60% of stems). Aboveground carbon stocks were negatively related to mean annual temperature, with forests at the warm end of the temperature range storing approximately half the amount of carbon as forests at the cool end of the temperature range. This may reflect thermal constraints on tree growth detected through other plot networks and physiological studies. Through common protocols and careful sampling design, the Ausplots Forest Monitoring Network will facilitate the integration of tall eucalypt forests into established global forest monitoring initiatives. In the context of projections of rapidly warming and drying climates in Australia, this plot network will enable detection of links between climate and growth, mortality and carbon dynamics of eucalypt forests.     

PCR-Free Enrichment of Mitochondrial DNA from Human Blood and Cell Lines for High Quality Next-Generation DNA Sequencing

Recent advances in sequencing technology allow for accurate detection of mitochondrial sequence variants, even those in low abundance at heteroplasmic sites. Considerable sequencing cost savings can be achieved by enriching samples for mitochondrial (relative to nuclear) DNA. Reduction in nuclear DNA (nDNA) content can also help to avoid false positive variants resulting from nuclear mitochondrial sequences (numts). We isolate intact mitochondrial organelles from both human cell lines and blood components using two separate methods: a magnetic bead binding protocol and differential centrifugation. DNA is extracted and further enriched for mitochondrial DNA (mtDNA) by an enzyme digest. Only 1 ng of the purified DNA is necessary for library preparation and next generation sequence (NGS) analysis. Enrichment methods are assessed and compared using mtDNA (versus nDNA) content as a metric, measured by using real-time quantitative PCR and NGS read analysis. Among the various strategies examined, the optimal is differential centrifugation isolation followed by exonuclease digest. This strategy yields >35% mtDNA reads in blood and cell lines, which corresponds to hundreds-fold enrichment over baseline. The strategy also avoids false variant calls that, as we show, can be induced by the long-range PCR approaches that are the current standard in enrichment procedures. This optimization procedure allows mtDNA enrichment for efficient and accurate massively parallel sequencing, enabling NGS from samples with small amounts of starting material. This will decrease costs by increasing the number of samples that may be multiplexed, ultimately facilitating efforts to better understand mitochondria-related diseases.     

A Multivariate Genome-Wide Association Analysis of 10 LDL Subfractions,and Their Response to Statin Treatment,in 1868 Caucasians

We conducted a genome-wide association analysis of 7 subfractions of low density lipoproteins (LDLs) and 3 subfractions of intermediate density lipoproteins (IDLs) measured by gradient gel electrophoresis, and their response to statin treatment, in 1868 individuals of European ancestry from the Pharmacogenomics and Risk of Cardiovascular Disease study. Our analyses identified four previously-implicated loci (SORT1, APOE, LPA, and CETP) as containing variants that are very strongly associated with lipoprotein subfractions (log₁₀Bayes Factor > 15). Subsequent conditional analyses suggest that three of these (APOE, LPA and CETP) likely harbor multiple independently associated SNPs. Further, while different variants typically showed different characteristic patterns of association with combinations of subfractions, the two SNPs in CETP show strikingly similar patterns - both in our original data and in a replication cohort - consistent with a common underlying molecular mechanism. Notably, the CETP variants are very strongly associated with LDL subfractions, despite showing no association with total LDLs in our study, illustrating the potential value of the more detailed phenotypic measurements. In contrast with these strong subfraction associations, genetic association analysis of subfraction response to statins showed much weaker signals (none exceeding log₁₀Bayes Factor of 6). However, two SNPs (in APOE and LPA) previously-reported to be associated with LDL statin response do show some modest evidence for association in our data, and the subfraction response proles at the LPA SNP are consistent with the LPA association, with response likely being due primarily to resistance of Lp(a) particles to statin therapy. An additional important feature of our analysis is that, unlike most previous analyses of multiple related phenotypes, we analyzed the subfractions jointly, rather than one at a time. Comparisons of our multivariate analyses with standard univariate analyses demonstrate that multivariate analyses can substantially increase power to detect associations. Software implementing our multivariate analysis methods is available at http://stephenslab.uchicago.edu/software.html.     

Longitudinal Neurostimulation in Older Adults Improves Working Memory

An increasing concern affecting a growing aging population is working memory (WM) decline. Consequently, there is great interest in improving or stabilizing WM, which drives expanded use of brain training exercises. Such regimens generally result in temporary WM benefits to the trained tasks but minimal transfer of benefit to untrained tasks. Pairing training with neurostimulation may stabilize or improve WM performance by enhancing plasticity and strengthening WM-related cortical networks. We tested this possibility in healthy older adults. Participants received 10 sessions of sham (control) or active (anodal, 1.5 mA) tDCS to the right prefrontal, parietal, or prefrontal/parietal (alternating) cortices. After ten minutes of sham or active tDCS, participants performed verbal and visual WM training tasks. On the first, tenth, and follow-up sessions, participants performed transfer WM tasks including the spatial 2-back, Stroop, and digit span tasks. The results demonstrated that all groups benefited from WM training, as expected. However, at follow-up 1-month after training ended, only the participants in the active tDCS groups maintained significant improvement. Importantly, this pattern was observed for both trained and transfer tasks. These results demonstrate that tDCS-linked WM training can provide long-term benefits in maintaining cognitive training benefits and extending them to untrained tasks.     

Diversity and potential sources of microbiota associated with snow on western portions of the Greenland Ice Sheet

Snow overlays the majority of the Greenland Ice Sheet (GrIS). However, there is very little information available on the microbiological assemblages that are associated with this vast and climate‐sensitive landscape. In this study, the structure and diversity of snow microbial assemblages from two regions of the western GrIS ice margin were investigated through the sequencing of small subunit ribosomal RNA genes. The origins of the microbiota were investigated by examining correlations to molecular data obtained from marine, soil, freshwater and atmospheric environments and geochemical analytes measured in the snow. Snow was found to contain a diverse assemblage of bacteria (Alphaproteobacteria, Betaproteobacteria and Gammaproteobacteria) and eukarya (Alveolata, Fungi, Stramenopiles and Chloroplastida). Phylotypes related to archaeal Thaumarchaeota and Euryarchaeota phyla were also identified. The snow microbial assemblages were more similar to communities characterized in soil than to those documented in marine ecosystems. Despite this, the chemical composition of snow samples was consistent with a marine contribution, and strong correlations existed between bacterial beta diversity and the concentration of Na⁺ and Cl^?. These results suggest that surface snow from western regions of Greenland contains exogenous microbiota that were likely aerosolized from more distant soil sources, transported in the atmosphere and co‐precipitated with the snow.