Collimated blue light generation in rubidium vapor

We describe an experiment for generating and characterizing a beam of collimated blue light (CBL) in a rubidium vapor.Two low-power, grating-feedback diode lasers, operating at 780.2 nm (5S3/2 → 5D5/2) and 776.0 nm (5P3/2 → 5D5/2), respectively, provide step-wise excitation to the 5D excited state in rubidium. Under the right experimental conditions, cascade decay through the 6P excited state will yield a collimated blue (420-nm) beam of light with high temporal and spatial coherence. We investigate the production of a blue beam under a variety of experimental conditions and characterize the spatial coherence and spectral characteristics. This experiment provides advanced undergraduate students with a unique opportunity to investigate nonlinear optical phenomena in the laboratory and uses equipment that is commonly available in laboratories equipped to investigate diode-laser-based absorption spectroscopy in rubidium

Collimated blue light generation in rubidium vapor

We describe an experiment for generating and characterizing a beam of collimated blue light (CBL) in a rubidium vapor.Two low-power, grating-feedback diode lasers, operating at 780.2 nm (5S3/2 → 5D5/2) and 776.0 nm (5P3/2 → 5D5/2), respectively, provide step-wise excitation to the 5D excited state in rubidium. Under the right experimental conditions, cascade decay through the 6P excited state will yield a collimated blue (420-nm) beam of light with high temporal and spatial coherence. We investigate the production of a blue beam under a variety of experimental conditions and characterize the spatial coherence and spectral characteristics. This experiment provides advanced undergraduate students with a unique opportunity to investigate nonlinear optical phenomena in the laboratory and uses equipment that is commonly available in laboratories equipped to investigate diode-laser-based absorption spectroscopy in rubidium

Modulation of γ-Secretase Activity by Multiple Enzyme-Substrate Interactions: Implications in Pathogenesis of Alzheimer's Disease

BACKGROUND: We describe molecular processes that can facilitate pathogenesis of Alzheimer's disease (AD) by analyzing the catalytic cycle of a membrane-imbedded protease γ-secretase, from the initial interaction with its C99 substrate to the final release of toxic Aβ peptides. RESULTS: The C-terminal AICD fragment is cleaved first in a pre-steady-state burst. The lowest Aβ42/Aβ40 ratio is observed in pre-steady-state when Aβ40 is the dominant product. Aβ42 is produced after Aβ40, and therefore Aβ42 is not a precursor for Aβ40. The longer more hydrophobic Aβ products gradually accumulate with multiple catalytic turnovers as a result of interrupted catalytic cycles. Saturation of γ-secretase with its C99 substrate leads to 30% decrease in Aβ40 with concomitant increase in the longer Aβ products and Aβ42/Aβ40 ratio. To different degree the same changes in Aβ products can be observed with two mutations that lead to an early onset of AD, ΔE9 and G384A. Four different lines of evidence show that γ-secretase can bind and cleave multiple substrate molecules in one catalytic turnover. Consequently depending on its concentration, NotchΔE substrate can activate or inhibit γ-secretase activity on C99 substrate. Multiple C99 molecules bound to γ-secretase can affect processive cleavages of the nascent Aβ catalytic intermediates and facilitate their premature release as the toxic membrane-imbedded Aβ-bundles. CONCLUSIONS: Gradual saturation of γ-secretase with its substrate can be the pathogenic process in different alleged causes of AD. Thus, competitive inhibitors of γ-secretase offer the best chance for a successful therapy, while the noncompetitive inhibitors could even facilitate development of the disease by inducing enzyme saturation at otherwise sub-saturating substrate. Membrane-imbedded Aβ-bundles generated by γ-secretase could be neurotoxic and thus crucial for our understanding of the amyloid hypothesis and AD pathogenesis

An evaluation of the self-assembly enhancing properties of cell-derived hexameric amyloid-β

A key hallmark of Alzheimer’s disease is the extracellular deposition of amyloid plaques composed primarily of the amyloidogenic amyloid-β (Aβ) peptide. The Aβ peptide is a product of sequential cleavage of the Amyloid Precursor Protein, the first step of which gives rise to a C-terminal Fragment (C99). Cleavage of C99 by γ-secretase activity releases Aβ of several lengths and the Aβ42 isoform in particular has been identified as being neurotoxic. The misfolding of Aβ leads to subsequent amyloid fibril formation by nucleated polymerisation. This requires an initial and critical nucleus for self-assembly. Here, we identify and characterise the composition and self-assembly properties of cell-derived hexameric Aβ42 and show its assembly enhancing properties which are dependent on the Aβ monomer availability. Identification of nucleating assemblies that contribute to self-assembly in this way may serve as therapeutic targets to prevent the formation of toxic oligomers