A murine model of variant late infantile ceroid lipofuscinosis recapitulates behavioral and pathological phenotypes of human disease.

Neuronal ceroid lipofuscinoses (NCLs; also known collectively as Batten Disease) are a family of autosomal recessive lysosomal storage disorders. Mutations in as many as 13 genes give rise to ∼10 variants of NCL, all with overlapping clinical symptomatology including visual impairment, motor and cognitive dysfunction, seizures, and premature death. Mutations in CLN6 result in both a variant late infantile onset neuronal ceroid lipofuscinosis (vLINCL) as well as an adult-onset form of the disease called Type A Kufs. CLN6 is a non-glycosylated membrane protein of unknown function localized to the endoplasmic reticulum (ER). In this study, we perform a detailed characterization of a naturally occurring Cln6 mutant (Cln6(nclf)) mouse line to validate its utility for translational research. We demonstrate that this Cln6(nclf) mutation leads to deficits in motor coordination, vision, memory, and learning. Pathologically, we demonstrate loss of neurons within specific subregions and lamina of the cortex that correlate to behavioral phenotypes. As in other NCL models, this model displays selective loss of GABAergic interneuron sub-populations in the cortex and the hippocampus with profound, early-onset glial activation. Finally, we demonstrate a novel deficit in memory and learning, including a dramatic reduction in dendritic spine density in the cerebral cortex, which suggests a reduction in synaptic strength following disruption in CLN6. Together, these findings highlight the behavioral and pathological similarities between the Cln6(nclf) mouse model and human NCL patients, validating this model as a reliable format for screening potential therapeutics

Flow Resistance Dynamics in Step-pool Channels: 2. Partitioning Between Grain, Spill, and Woody Debris Resistance

In step-pool stream channels, flow resistance is created primarily by bed sediments, spill over step-pool bed forms, and large woody debris (LWD). In order to measure resistance partitioning between grains, steps, and LWD in step-pool channels we completed laboratory flume runs in which total resistance was measured with and without grains and steps, with various LWD configurations, and at multiple slopes and discharges. Tests of additive approaches to resistance partitioning found that partitioning estimates are highly sensitive to the order in which components are calculated and that such approaches inflate the values of difficult-to-measure components that are calculated by subtraction from measured components. This effect is especially significant where interactions between roughness features create synergistic increases in resistance such that total resistance measured for combinations of resistance components greatly exceeds the sum of those components measured separately. LWD contributes large proportions of total resistance by creating form drag on individual pieces and by increasing the spill resistance effect of steps. The combined effect of LWD and spill over steps was found to dominate total resistance, whereas grain roughness on step treads was a small component of total resistance. The relative contributions of grain, spill, and woody debris resistance were strongly influenced by discharge and to a lesser extent by LWD density. Grain resistance values based on published formulas and debris resistance values calculated using a cylinder drag approach typically underestimated analogous flume-derived values, further illustrating sources of error in partitioning methods and the importance of accounting for interaction effects between resistance components

Pain Experiences and Their Relation to Opioid Misuse Risk and Emotion Dysregulation

Pain is a complex, multidimensional experience but often is measured as a unidimensional experience. This study aimed to separately assess the sensory and affective components of pain and identify their relations to important pain-related outcomes, particularly in terms of opioid misuse risk and emotion dysregulation among patients with chronic pain receiving treatment in Appalachia. Two hundred and twelve patients presenting to a multidisciplinary pain center completed the Difficulties in Emotion Regulation Scale (DERS-18), Screener and Opioid Assessment for Patients with Pain—Revised (SOAPP-R), and short-form McGill Pain Questionnaire (SF-MPQ). The sensory experience of pain was unrelated to emotion dysregulation (r = 0.06, p = 0.57) and weakly related to opioid misuse risk (r = 0.182, p \u3c 0.05). In contrast, the affective experience of pain was moderately related to emotion dysregulation (r = 0.217, p \u3c 0.05) and strongly related to opioid misuse risk (r = 0.37, p \u3c 0.01). In addition, emotion dysregulation predicted variance in opioid misuse risk above and beyond the affective and sensory experiences of pain ((b = 0.693, p \u3c 0.001). The results suggest patients with a strong affective experience versus sensory experience of pain and challenges with emotion regulation may require a more comprehensive intervention to address these underlying components in order to reduce their risk of misusing opioid medications

Solvent effects on Grubbs’ pre-catalyst initiation rates

Initiation rates for Grubbs and Grubbs-Hoveyda second generation pre-catalysts have been measured accurately in a range of solvents. Solvatochromic fitting reveals different dependencies on key solvent parameters for the two pre-catalysts, consistent with different mechanisms by which the Grubbs and Grubbs-Hoveyda pre-catalysts initiate

What is the initiation step of the Grubbs-Hoveyda olefin metathesis catalyst?

Density function theory calculations reveal that the Grubbs-Hoveyda olefin metathesis pre-catalyst is activated by the formation of a complex in which the incoming alkene substrate and outgoing alkoxy ligand are both clearly associated with the ruthenium centre. The computed energies for reaction are in good agreement with the experimental values, reported here

The electrophilic fluorination of enol esters using SelectFluor : a polar two-electron process

The reaction of enol esters with SelectFluor is facile and leads to the corresponding α-fluoroketones under mild conditions and, as a result, this route is commonly employed for the synthesis of medicinally important compounds such as fluorinated steroids. However, despite the use of this methodology in synthesis, the mechanism of this reaction and the influence of structure on reactivity are unclear. We present a rigorous mechanistic study of the fluorination of these substrates, informed primarily by detailed and robust kinetic experiments. The results of this study implicate a polar two-electron process via an oxygen-stabilised carbenium species, rather than a single-electron process involving radical intermediates. The structure/reactivity relationships revealed here will assist synthetic chemists in deploying this type of methodology in the syntheses of α-fluoroketone

A synthesis-enabled relative stereochemical assignment of the C1-C28 region of hemicalide.

Through synthesising both candidate diastereomers of a model C1-C28 fragment of the potent cytotoxic marine polyketide hemicalide, an assignment of the relative configuration between the C1-C15 and C16-C26 regions has been achieved. By detailed NMR comparisons with the natural product, the relative stereochemistry between these two 1,6-related stereoclusters is elucidated as 13,18-syn rather than the previously proposed 13,18-anti relationship. A flexible and modular strategy using an advanced C1-C28 ketone fragment 22 is outlined to elucidate the remaining stereochemical features and achieve a total synthesis

Ribosomal Proteins RPS11 and RPS20, Two Stress-Response Markers of Glioblastoma Stem Cells, Are Novel Predictors of Poor Prognosis in Glioblastoma Patients.

Glioblastoma stem cells (GSC) co-exhibiting a tumor-initiating capacity and a radio-chemoresistant phenotype, are a compelling cell model for explaining tumor recurrence. We have previously characterized patient-derived, treatment-resistant GSC clones (TRGC) that survived radiochemotherapy. Compared to glucose-dependent, treatment-sensitive GSC clones (TSGC), TRGC exhibited reduced glucose dependence that favor the fatty acid oxidation pathway as their energy source. Using comparative genome-wide transcriptome analysis, a series of defense signatures associated with TRGC survival were identified and verified by siRNA-based gene knockdown experiments that led to loss of cell integrity. In this study, we investigate the prognostic value of defense signatures in glioblastoma (GBM) patients using gene expression analysis with Probeset Analyzer (131 GBM) and The Cancer Genome Atlas (TCGA) data, and protein expression with a tissue microarray (50 GBM), yielding the first TRGC-derived prognostic biomarkers for GBM patients. Ribosomal protein S11 (RPS11), RPS20, individually and together, consistently predicted poor survival of newly diagnosed primary GBM tumors when overexpressed at the RNA or protein level [RPS11: Hazard Ratio (HR) = 11.5, p<0.001; RPS20: HR = 4.5, p = 0.03; RPS11+RPS20: HR = 17.99, p = 0.001]. The prognostic significance of RPS11 and RPS20 was further supported by whole tissue section RPS11 immunostaining (27 GBM; HR = 4.05, p = 0.01) and TCGA gene expression data (578 primary GBM; RPS11: HR = 1.19, p = 0.06; RPS20: HR = 1.25, p = 0.02; RPS11+RPS20: HR = 1.43, p = 0.01). Moreover, tumors that exhibited unmethylated O-6-methylguanine-DNA methyltransferase (MGMT) or wild-type isocitrate dehydrogenase 1 (IDH1) were associated with higher RPS11 expression levels [corr (IDH1, RPS11) = 0.64, p = 0.03); [corr (MGMT, RPS11) = 0.52, p = 0.04]. These data indicate that increased expression of RPS11 and RPS20 predicts shorter patient survival. The study also suggests that TRGC are clinically relevant cells that represent resistant tumorigenic clones from patient tumors and that their properties, at least in part, are reflected in poor-prognosis GBM. The screening of TRGC signatures may represent a novel alternative strategy for identifying new prognostic biomarkers

Stratigraphy, Radiocarbon Dating and Culture History of Charlie Lake Cave, British Columbia

Three seasons of fieldwork at Charlie Lake Cave, British Columbia, have revealed a sequence of stratified deposits that spans the Late Pleistocene and entire Holocene. Analyses of sediments, radiocarbon dates, faunal remains, and artifacts show that the site was first occupied by people at about 10 500 B.P., when local environments were more open than today. By 9500 B.P., boreal forest had moved into the area, and human use of the site was minimal until about 7000 B.P., when a brief occupation of the site probably included a human burial. Use of the site intensified after about 4500 B.P., possibly because the cave became more accessible. The site was used both as a residential base camp and as a more temporary hunting station or lookout.   Article Summary by Jonathan C. Driver, May 2015   After we had completed our excavations in 1991 we decided that we should focus on writing up what we had excavated, and not undertake further excavations at the site. This paper was written to summarize our state of knowledge about the archaeological remains at the site, and focused on describing the stone tools, the overall stratigraphy, and the dating. The stratigraphic summary replaced earlier work based on the 1983 excavations, and we were able to refine our dating of the site as a result of more radiocarbon dates. The different cultural periods were based mainly on the work done by Martin Handly for his M.A. thesis at Trent University. The long list of authors reflects the need for a team approach to archaeological work, and include the project directors (Knut Fladmark and Jon Driver), stone tool analysis and development of the cultural sequence (Martin Handly), animal bone analysis (Randall Preston and Jon Driver), sediment analysis (Greg Sullivan and Knut Fladmark), and radiocarbon dating (Erle Nelson). The most important aspect of the site is that it preserves a very rare record of humanly made artifacts dating from the end of the last ice age (at least 10,500 BC) to very recent times. The many layers at the site allow us to separate the different cultural periods. Good preservation of bone allowed us to submit radiocarbon dates that provide approximate ages for the various cultures that used the site. The artifacts that exhibit the most change through time are projectile points – the sharp stone tips for spears, darts and arrows. In much of western Canada it is difficult to date archaeological sites, because many of them are found in shallow soils where radiocarbon dating is difficult for two reasons. First, animal bone is often not preserved due to the acidic nature of the soils. Second, although charcoal is often found, it cannot be reliably associated with human activity, because natural forest fires also produce charcoal. As a result, archaeologists look at the style of the projectile points to assign approximate ages. Tse’K’wa provides an opportunity to link artifacts of different styles to radiocarbon dates in a site with many distinct layers. So Tse’K’wa is a foundation for understanding the sequence of different cultures in the region. The article also discusses the possible early presence of microblade technology. Microblades are the most efficient way of producing a cutting edge when the base technology is chipped stone. A small piece of high-quality raw material (known as the core) is shaped in such a way that numerous parallel-sided slivers of stone can be removed. These “microblades” can then be hafted in wood or antler to form knives or arrow barbs. The concept is rather like our utility knives that have replaceable blades. This technology allows people to carry small quantities of high quality stone with them, ensuring that they always have a sharp blade available. Not all archaeologists agree that the early microblade core from Tse’K’wa is part of this technology, because it doesn’t conform to the classic methods of core manufacture. However, we argue that evidence for the removal of microblades is very obvious, and the lack of classic core preparation is because of the tabular nature of the raw material. The paper also introduces some information about animal bones, including the raven burials, evidence for environmental change, and the presence of collared lemming. These topics were subsequently explored in more detail in other papers

in silico verification and parallel reaction monitoring prevalidation of potential prostate cancer biomarkers

Purpose: Targeted proteomics of potential biomarkers is often challenging. Hence, we developed an intermediate workflow to streamline potential urinary biomarkers of prostate cancer (PCa). Materials & methods: Using previously discovered potential PCa biomarkers; we selected proteotypic peptides for targeted validation. Preliminary in silico immunohistochemical and single reaction monitoring (SRM) verification was performed. Successful PTPs were then prevalidated using parallel reaction monitoring (PRM) and reconfirmed in 15 publicly available databases. Results: Stringency-based targetable potential biomarkers were shortlisted following in silico screening. PRM reveals top 12 potential biomarkers including the top ranking seven in silico verification-based biomarkers. Database reconfirmation showed differential expression between PCa and benign/normal prostatic urine samples. Conclusion: The pragmatic penultimate screening step, described herein, would immensely improve targeted proteomics validation of ..