Strain-dependent variation in the early transcriptional response to CNS injury using a cortical explant system

<p>Abstract</p> <p>Background</p> <p>While it is clear that inbred strains of mice have variations in immunological responsiveness, the influence of genetic background following tissue damage in the central nervous system is not fully understood. A cortical explant system was employed as a model for injury to determine whether the immediate transcriptional response to tissue resection revealed differences among three mouse strains.</p> <p>Methods</p> <p>Immunological mRNAs were measured in cerebral cortex from SJL/J, C57BL/6J, and BALB/cJ mice using real time RT-PCR. Freshly isolated cortical tissue and cortical sections incubated in explant medium were examined. Levels of mRNA, normalized to β-actin, were compared using one way analysis of variance with pooled samples from each mouse strain.</p> <p>Results</p> <p>In freshly isolated cerebral cortex, transcript levels of many pro-inflammatory mediators were not significantly different among the strains or too low for comparison. Constitutive, baseline amounts of CD74 and antisecretory factor (ASF) mRNAs, however, were higher in SJL/J and C57BL/6J, respectively. When sections of cortical tissue were incubated in explant medium, increased message for a number of pro-inflammatory cytokines and chemokines occurred within five hours. Message for chemokines, IL-1α, and COX-2 transcripts were higher in C57BL/6J cortical explants relative to SJL/J and BALB/cJ. IL-1β, IL-12/23 p40, and TNF-α were lower in BALB/cJ explants relative to SJL/J and C57BL/6J. Similar to observations in freshly isolated cortex, CD74 mRNA remained higher in SJL/J explants. The ASF mRNA in SJL/J explants, however, was now lower than levels in both C57BL/6J and BALB/cJ explants.</p> <p>Conclusions</p> <p>The short-term cortical explant model employed in this study provides a basic approach to evaluate an early transcriptional response to neurological damage, and can identify expression differences in genes that are influenced by genetic background.</p

Progressive changes in microglia and macrophages in spinal cord and peripheral nerve in the transgenic rat model of amyotrophic lateral sclerosis

<p>Abstract</p> <p>Background</p> <p>The role of neuroinflammation in motor neuron death of amyotrophic lateral sclerosis (ALS) is unclear. The human mutant superoxide dismutase-1 (hmSOD1)-expressing murine transgenic model of ALS has provided some insight into changes in microglia activity during disease progression. The purpose of this study was to gain further knowledge by characterizing the immunological changes during disease progression in the spinal cord and peripheral nerve using the more recently developed hmSOD1 rat transgenic model of ALS.</p> <p>Methods</p> <p>Using immunohistochemistry, the extent and intensity of tissue CD11b expression in spinal cord, lumbar nerve roots, and sciatic nerve were evaluated in hmSOD1 rats that were pre-clinical, at clinical onset, and near disease end-stage. Changes in CD11b expression were compared to the detection of MHC class II and CD68 microglial activation markers in the ventral horn of the spinal cord, as well as to the changes in astrocytic GFAP expression.</p> <p>Results</p> <p>Our study reveals an accumulation of microglia/macrophages both in the spinal cord and peripheral nerve prior to clinical onset based on CD11b tissue expression. The microglia formed focal aggregates in the ventral horn and became more widespread as the disease progressed. Hypertrophic astrocytes were not prominent in the ventral horn until after clinical onset, and the enhancement of GFAP did not have a strong correlation to increased CD11b expression. Detection of MHC class II and CD68 expression was found in the ventral horn only after clinical onset. The macrophages in the ventral nerve root and sciatic nerve of hmSOD1 rats were observed encircling axons.</p> <p>Conclusions</p> <p>These findings describe for the first time in the hmSOD1 rat transgenic model of ALS that enhancement of microglia/macrophage activity occurs pre-clinically both in the peripheral nerve and in the spinal cord. CD11b expression is shown to be a superior indicator for early immunological changes compared to other microglia activation markers and astrogliosis. Furthermore, we suggest that the early activity of microglia/macrophages is involved in the early phase of motor neuron degeneration and propose that studies involving immunomodulation in hmSOD1transgenic models need to consider effects on macrophages in peripheral nerves as well as to microglia in the spinal cord.</p

AN IMPROVED ESTIMATOR FOR ASSESSING THE MEASURE OF AGREEMENT WITH A GOLD STANDARD

St. Laurent (1998, Biometrics 54, 537-545) developed a measure of agreement for method comparison studies in which an approximate method of measurement is compared to a gold standard method of measurement. The measure of agreement proposed was shown to be related to a population intraclass correlation coefficient. This paper develops a family of estimators for the measure of agreement based on pivotal quantities. A blend of two particular members of the family is suggested as an estimator itself. In general, this estimator outperforms the maximum likelihood estimator in terms of bias and mean-squared error

Australia’s first trap-and-haul fishway: Nerang River, Queensland

Australia’s first trap-and-haul fishway was built on the Nerang River in southeast Queensland during enlargement of the Hinze Dam. In gaining government approval for the project, the constructing alliance (HDA) concluded that providing upstream fish passage would have significant environmental benefits by conserving upstream native fish communities and encouraging recruitment into the reservoir’s recreational fishery. But downstream passage, other than modifications to protect fish emigrating during spill events, could not be justified. Severely constrained outflows from the dam and the impacts of downstream urban development contrast starkly with upstream habitat conditions. North American design approaches were adapted to suit the river’s fishes and streamflows. Challenges for fish passage at the site included the marked restriction of downstream river flows and the location of the flow-release point 300m from the spillway. A barrier weir was built to prevent upstream migrants bypassing the fishway entrance during spillway flows. HDA developed a trap-and-haul system to collect migrating fish at the weir and transport them by tanker to multiple upstream release areas. This system provides flexibility to accommodate varying fish biomass; ability to operate over a range of flows up to 20-year ARI events; facilities for sorting, data collection and removal of pest species; and capacity to limit predation mortalities. Fishway performance studies led to redesign of the entrance vee-trap and other modifications. To 2017, 55,590 fish from 27 large and small-bodied species used the fishway, together with 8 turtles. Fish of three pest species were identified in the sorting facility and removed to prevent their entry to the reservoir. This paper covers the project life cycle for Australia’s first trap-and-haul fishway including the basis for selecting the fishway type, design and construction. Fishway performance studies and results from ongoing operation, including the lessons learned and the improvements made, are also discussed

Proof of concept for an innovative pump fishway design to move fish upstream over dams

Reversing worldwide declines in freshwater fish while making sustainable use of water resources will require effective and economical fishways to restore fish migrations. Mitigation of barrier effects at dams and weirs is too often impeded by poor fishway performance and high costs, so that many fish migrations continue to be obstructed. Improved and less-costly designs are urgently needed. Our innovative pump fishway concept combines fish-behaviour insights, proved fishways techniques and aquaculture’s pumping methods for safe upstream transport of living fish. We ran a series of experimental trials using several scale-model fishway designs with young, hatchery-bred fish. Our horizontal-cylinder design successfully combined volitional-passage functions of existing fishways with non-volitional transport in a conduit carrying pumped water. Several key principles of fish behaviour in fishways led to design improvements: disturbed fish often seek refuge at depth; fishes’ escape reactions strongly motivate swimming into flows; and curved structures aid passage by reducing delays. Replicated trials finally produced an average of 98% successful passage, within brief cycling periods. The pump fishway concept offers potential for effective upstream fish passage at new and existing sites \u3e~2m high, with low construction and maintenance costs and highly adaptable operation in variable flow regimes. Development beyond the concept-trial phase is now a priority

Planning and establishment of a high throughput screening site

In 1996 and 1997, Glaxo Wellcome's US Research division planned and established their second generation research strategy. An important aspect of the strategy entailed development of two automated screening sites in Biochemistry in Research Triangle Park, NC. Development of the new operations required many decisions to be made very quickly, including automated process design, system selection and site preparation. Descriptions of the decision made in the development of one of the screening sites are presented in this paper

Gene Ontology Analysis of Pairwise Genetic Associations in Two Genome-Wide Studies of Sporadic ALS

It is increasingly clear that common human diseases have a complex genetic architecture characterized by both additive and nonadditive genetic effects. The goal of the present study was to determine whether patterns of both additive and nonadditive genetic associations aggregate in specific functional groups as defined by the Gene Ontology (GO)

Combined fluorescence and reflectance spectroscopy for in vivo quantification of cancer biomarkers in low - and high- grade glioma surgery

Biomarkers are indicators of biological processes and hold promise for the diagnosis and treatment of disease. Gliomas represent a heterogeneous group of brain tumors with marked intra- and inter-tumor variability. The extent of surgical resection is a significant factor influencing post-surgical recurrence and prognosis. Here, we used fluorescence and reflectance spectral signatures for in vivo quantification of multiple biomarkers during glioma surgery, with fluorescence contrast provided by exogenously-induced protoporphyrin IX (PpIX) following administration of 5-aminolevulinic acid. We performed light-transport modeling to quantify multiple biomarkers indicative of tumor biological processes, including the local concentration of PpIX and associated photoproducts, total hemoglobin concentration, oxygen saturation, and optical scattering parameters.We developed a diagnostic algorithm for intra-operative tissue delineation that accounts for the combined tumor-specific predictive capabilities of these quantitative biomarkers. Tumor tissue delineation achieved accuracies of up to 94% (specificity=94%, sensitivity=94%) across a range of glioma histologies beyond current state-of-the-art optical approaches, including state-of-the-art fluorescence image guidance. This multiple biomarker strategy opens the door to optical methods for surgical guidance that use quantification of well-established neoplastic processes. Future work would seek to validate the predictive power of this proof-of-concept study in a separate larger cohort of patients

Multifactor dimensionality reduction for graphics processing units enables genome-wide testing of epistasis in sporadic ALS

Motivation: Epistasis, the presence of gene–gene interactions, has been hypothesized to be at the root of many common human diseases, but current genome-wide association studies largely ignore its role. Multifactor dimensionality reduction (MDR) is a powerful model-free method for detecting epistatic relationships between genes, but computational costs have made its application to genome-wide data difficult. Graphics processing units (GPUs), the hardware responsible for rendering computer games, are powerful parallel processors. Using GPUs to run MDR on a genome-wide dataset allows for statistically rigorous testing of epistasis