65 research outputs found
Microdissection: A method developed to investigate mechanisms involved in transmissible spongiform encephalopathy pathogenesis
BACKGROUND: The transmissible spongiform encephalopathies (TSEs) are a group of neurodegenerative diseases affecting both human and animals. The neuroanatomical changes which occur in the central nervous system (CNS) of TSE infected animals include vacuolation, gliosis, neuronal loss and the deposition of a disease specific protein, PrP(Sc). Experimental murine models of scrapie, a TSE of sheep, have revealed that pathology may be confined to specific brain areas with targeting of particular neuronal subsets depending on route of injection and scrapie isolate. To assess the biochemical changes which are taking place in these targeted areas it was necessary to develop a reliable sampling procedure (microdissection) which could be used for a variety of tests such as western blotting and magnetic resonance spectroscopy. METHODS: The method described is for the microdissection of murine brains. To assess the usefulness of this dissection technique for producing similar sample types for analysis by various down-stream biochemical techniques, the areas dissected were analysed for PrP(Sc )by western blotting and compared to immunocytochemical (ICC) techniques. RESULTS: Results show that the method generates samples yielding a consistent protein content which can be analysed for PrP(Sc). The areas in which PrP(Sc )is found by western blotting compares well with localisation visualised by immunocytochemistry. CONCLUSION: The microdisssection method described can be used to generate samples suitable for a range of biochemical techniques. Using these samples a range of assays can be carried out which will help to elucidate the molecular and cellular mechanisms underlying TSE pathogenesis. The method would also be useful for any study requiring the investigation of discrete areas within the murine brain
Protease-Sensitive Conformers in Broad Spectrum of Distinct PrPSc Structures in Sporadic Creutzfeldt-Jakob Disease Are Indicator of Progression Rate
The origin, range, and structure of prions causing the most common human prion disease, sporadic Creutzfeldt-Jakob disease (sCJD), are largely unknown. To investigate the molecular mechanism responsible for the broad phenotypic variability of sCJD, we analyzed the conformational characteristics of protease-sensitive and protease-resistant fractions of the pathogenic prion protein (PrPSc) using novel conformational methods derived from a conformation-dependent immunoassay (CDI). In 46 brains of patients homozygous for polymorphisms in the PRNP gene and exhibiting either Type 1 or Type 2 western blot pattern of the PrPSc, we identified an extensive array of PrPSc structures that differ in protease sensitivity, display of critical domains, and conformational stability. Surprisingly, in sCJD cases homozygous for methionine or valine at codon 129 of the PRNP gene, the concentration and stability of protease-sensitive conformers of PrPSc correlated with progression rate of the disease. These data indicate that sCJD brains exhibit a wide spectrum of PrPSc structural states, and accordingly argue for a broad spectrum of prion strains coding for different phenotypes. The link between disease duration, levels, and stability of protease-sensitive conformers of PrPSc suggests that these conformers play an important role in the pathogenesis of sCJD
More stories on Th17 cells
For more than two decades, immunologists have been using the so-called Th1/Th2 paradigm to explain most of the phenomena related to adaptive immunity. the Th1/Th2 paradigm implied the existence of two different, mutually regulated, CD4(+) T helper subsets: Th1 cells, driving cell-mediated immune responses involved in tissue damage and fighting infection against intracellular parasites; and Th2 cells that mediate IgE production and are particularly involved in eosinophilic inflammation, allergy and clearance of helminthic infections. A third member of the T helper set, IL-17-producing CD4(+) T cells, now called Th17 cells, was recently described as a distinct lineage that does not share developmental pathways with either Th1 or Th2 cells. the Th17 subset has been linked to autoimmune disorders, being able to produce IL-17, IL-17F and IL-21 among other inflammatory cytokines. Interestingly, it has been reported that there is not only a cross-regulation among Th1, Th2 and Th17 effector cells but there is also a dichotomy in the generation of Th17 and T regulatory cells. Therefore, Treg and Th17 effector cells arise in a mutually exclusive fashion, depending on whether they are activated in the presence of TGF-beta or TGF-beta plus inflammatory cytokines such as IL-6. This review will address the discovery of the Th17 cells, and recent progress on their development and regulation.Crohn's and Colitis Foundation of AmericaNIHLa Jolla Inst Allergy & Immunol, La Jolla, CA 92037 USAUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilNIH: RO1 AI050265-06Web of Scienc
Observations of Ly Emitters at High Redshift
In this series of lectures, I review our observational understanding of
high- Ly emitters (LAEs) and relevant scientific topics. Since the
discovery of LAEs in the late 1990s, more than ten (one) thousand(s) of LAEs
have been identified photometrically (spectroscopically) at to . These large samples of LAEs are useful to address two major astrophysical
issues, galaxy formation and cosmic reionization. Statistical studies have
revealed the general picture of LAEs' physical properties: young stellar
populations, remarkable luminosity function evolutions, compact morphologies,
highly ionized inter-stellar media (ISM) with low metal/dust contents, low
masses of dark-matter halos. Typical LAEs represent low-mass high- galaxies,
high- analogs of dwarf galaxies, some of which are thought to be candidates
of population III galaxies. These observational studies have also pinpointed
rare bright Ly sources extended over kpc, dubbed
Ly blobs, whose physical origins are under debate. LAEs are used as
probes of cosmic reionization history through the Ly damping wing
absorption given by the neutral hydrogen of the inter-galactic medium (IGM),
which complement the cosmic microwave background radiation and 21cm
observations. The low-mass and highly-ionized population of LAEs can be major
sources of cosmic reionization. The budget of ionizing photons for cosmic
reionization has been constrained, although there remain large observational
uncertainties in the parameters. Beyond galaxy formation and cosmic
reionization, several new usages of LAEs for science frontiers have been
suggested such as the distribution of {\sc Hi} gas in the circum-galactic
medium and filaments of large-scale structures. On-going programs and future
telescope projects, such as JWST, ELTs, and SKA, will push the horizons of the
science frontiers.Comment: Lecture notes for `Lyman-alpha as an Astrophysical and Cosmological
Tool', Saas-Fee Advanced Course 46. Verhamme, A., North, P., Cantalupo, S., &
Atek, H. (eds.) --- 147 pages, 103 figures. Abstract abridged. Link to the
lecture program including the video recording and ppt files :
https://obswww.unige.ch/Courses/saas-fee-2016/program.cg
Shedding Light on the Galaxy Luminosity Function
From as early as the 1930s, astronomers have tried to quantify the
statistical nature of the evolution and large-scale structure of galaxies by
studying their luminosity distribution as a function of redshift - known as the
galaxy luminosity function (LF). Accurately constructing the LF remains a
popular and yet tricky pursuit in modern observational cosmology where the
presence of observational selection effects due to e.g. detection thresholds in
apparent magnitude, colour, surface brightness or some combination thereof can
render any given galaxy survey incomplete and thus introduce bias into the LF.
Over the last seventy years there have been numerous sophisticated
statistical approaches devised to tackle these issues; all have advantages --
but not one is perfect. This review takes a broad historical look at the key
statistical tools that have been developed over this period, discussing their
relative merits and highlighting any significant extensions and modifications.
In addition, the more generalised methods that have emerged within the last few
years are examined. These methods propose a more rigorous statistical framework
within which to determine the LF compared to some of the more traditional
methods. I also look at how photometric redshift estimations are being
incorporated into the LF methodology as well as considering the construction of
bivariate LFs. Finally, I review the ongoing development of completeness
estimators which test some of the fundamental assumptions going into LF
estimators and can be powerful probes of any residual systematic effects
inherent magnitude-redshift data.Comment: 95 pages, 23 figures, 3 tables. Now published in The Astronomy &
Astrophysics Review. This version: bring in line with A&AR format
requirements, also minor typo corrections made, additional citations and
higher rez images adde
Enzymatic formulation capable of degrading scrapie prion under mild digestion conditions
The prion agent is notoriously resistant to common proteases and conventional sterilisation procedures. The current methods known to destroy prion infectivity such as incineration, alkaline and thermal hydrolysis are harsh, destructive, environmentally polluting and potentially hazardous, thus limit their applications for decontamination of delicate medical and laboratory devices, remediation of prion contaminated environment and for processing animal by-products including specified risk materials and carcases. Therefore, an environmentally friendly, non-destructive enzymatic degradation approach is highly desirable. A feather-degrading Bacillus licheniformis N22 keratinase has been isolated which degraded scrapie prion to undetectable level of PrPSc signals as determined by Western Blot analysis. Prion infectivity was verified by ex vivo cell-based assay. An enzymatic formulation combining N22 keratinase and biosurfactant derived from Pseudomonas aeruginosa degraded PrPSc at 65°C in 10 min to undetectable level -. A time-course degradation analysis carried out at 50°C over 2 h revealed the progressive attenuation of PrPSc intensity. Test of residual infectivity by standard cell culture assay confirmed that the enzymatic formulation reduced PrPSc infectivity to undetectable levels as compared to cells challenged with untreated standard scrapie sheep prion (SSBP/1) (p-value = 0.008 at 95% confidence interval). This novel enzymatic formulation has significant potential application for prion decontamination in various environmentally friendly systems under mild treatment conditions
Prionen als Erreger der Creutzfeldt-Jakob-Erkrankungen und der bovinen spongiformen Enzephalopathie (BSE)
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