Prion Protein Is Decreased in Alzheimer's Brain and Inversely Correlates with BACE1 Activity,Amyloid-β Levels and Braak Stage

The cellular prion protein (PrP^C) has been implicated in the development of Alzheimer''s disease (AD). PrP^C decreases amyloid-β (Aβ) production, which is involved in AD pathogenesis, by inhibiting β-secretase (BACE1) activity. Contactin 5 (CNTN5) has also been implicated in the development of AD by a genome-wide association study. Here we measured PrP^C and CNTN5 in frontal cortex samples from 24 sporadic AD and 24 age-matched control brains and correlated the expression of these proteins with markers of AD. PrP^C was decreased in sporadic AD compared to controls (by 49%, p = 0.014) but there was no difference in CNTN5 between sporadic AD and controls (p = 0.217). PrP^C significantly inversely correlated with BACE1 activity (r_s = −0.358, p = 0.006), Aβ load (r_s = −0.456, p = 0.001), soluble Aβ (r_s = −0.283, p = 0.026) and insoluble Aβ (r_s = −0.353, p = 0.007) and PrP^C also significantly inversely correlated with the stage of disease, as indicated by Braak tangle stage (r_s = −0.377, p = 0.007). CNTN5 did not correlate with Aβ load (r_s = 0.040, p = 0.393), soluble Aβ (r_s = 0.113, p = 0.223) or insoluble Aβ (r_s = 0.169, p = 0.125). PrP^C was also measured in frontal cortex samples from 9 Down''s syndrome (DS) and 8 age-matched control brains. In contrast to sporadic AD, there was no difference in PrP^C in the DS brains compared to controls (p = 0.625). These data are consistent with a role for PrP^C in regulating Aβ production and indicate that brain PrP^C level may be important in influencing the onset and progression of sporadic AD.     

Scanning electron microscopic observations of early stages of phagocytosis of E. coli by human neutrophils

Summary Changes in surface morphology, as observed by scanning electron microscopy, appear rapidly when human polymorphonuclear neutrophils (PMN) are challenged with bacteria. Monolayers of PMN adhering to glass were incubated with opsonized E. coli from 5 sec to 10 min, and then fixed and prepared for SEM. As early as 5 sec after phagocytic challenge, E. coli are found in contact with PMN and in the process of engulfment into open cavities formed by lamellipodia. The shape of the mouth of the forming phagocytic vacuole is related to the orientation of bacteria during entry. Bacteria engulfed into early forming phagosomes are surrounded by a large open space between the bacteria and the phagosome wall. As phagocytosis proceeds, the space is reduced and the loose fit around the entering bacteria becomes tight. By 30 sec, bacteria may be completely internalized and by 1 min phagocytized E. coli are packed into bulging PMN. The observations reveal the variability and rapidity of the phagocytic response and confirm the presence of sensitive mechanisms for host defense by PMN.This work was supported by research grants from the University of North Carolina Research Council and the National Institutes of Health (A1 02430)     

Nitric Oxide and Cnidarian-Dinoflagellate Symbioses: Pieces of a Puzzle

The presence of nitric oxide synthase (NOS) activity is demonstratedin the tropical marine cnidarian Aiptasia pallida and in itssymbiotic dinoflagellate algae, Symbiodinium bermudense. Enzymeactivity was assayed by measuring the conversion of arginineto citrulline. Biochemical characterization of NOS from Aiptasiawas characterized with respect to cellular localization, substrateand cofactor requirements, inhibitors, and kinetics. In responseto acute temperature shock, anemones retracted their tentacles.Animals subjected to such stress had lower NOS activities thandid controls. Treatment with NOS inhibitors caused tentacularretraction, while treatment with the NOS substrate L-arginineinhibited this response to stress, as did treatment with NOdonors. These results provide a preliminary biochemical characterizationof, and suggest a functional significance for, NOS activityin anthozoan-algal symbiotic assemblages.     

Intensive Care Ethics in Evolution

The ethics of treating the seriously and critically ill have not been static throughout the ages. Twentieth century medicine has inherited from the nineteenth century a science which places an inappropriate weight on diagnosis over prognosis and management, combined with a seventeenth century duty to prolong life. However other earlier ethical traditions, both Hippocratic and Christian, respected both the limitations of medicine and emphasised the importance of prognosis. This paper outlines some of the historical precedents for the treatment of the critically ill, and also how the current paradigm limits clinical practice and causes ethical tensions. An understanding that other paradigms have been ethically acceptable in the past allows wider consideration and acceptance of alternatives for the future. However future alternatives will also have to address the role of technology, given its importance in this area of medicine.     

Dynamic Localization of Tat Protein Transport Machinery Components in Streptomyces coelicolor

The Tat pathway transports folded proteins across the bacterial cytoplasmic membrane and is a major route of protein export in the Streptomyces genus of bacteria. In this study, we have examined the localization of Tat components in the model organism Streptomyces coelicolor by constructing enhanced green fluorescent protein (eGFP) and mCherry fusions with the TatA, TatB, and TatC proteins. All three components colocalized dynamically in the vegetative hyphae, with foci of each tagged protein being prominent at the tips of emerging germ tubes and of the vegetative hyphae, suggesting that this may be a primary site of Tat secretion. Time-lapse imaging revealed that localization of the Tat components was highly dynamic during tip growth and again demonstrated a strong preference for apical sites in growing hyphae. During aerial hypha formation, TatA-eGFP and TatB-eGFP fusions relocalized to prespore compartments, indicating repositioning of Tat components during the Streptomyces life cycle.     

WIKI4, a Novel Inhibitor of Tankyrase and Wnt/?-Catenin Signaling

The Wnt/ß-catenin signaling pathway controls important cellular events during development and often contributes to disease when dysregulated. Using high throughput screening we have identified a new small molecule inhibitor of Wnt/ß-catenin signaling, WIKI4. WIKI4 inhibits expression of ß-catenin target genes and cellular responses to Wnt/ß-catenin signaling in cancer cell lines as well as in human embryonic stem cells. Furthermore, we demonstrate that WIKI4 mediates its effects on Wnt/ß-catenin signaling by inhibiting the enzymatic activity of TNKS2, a regulator of AXIN ubiquitylation and degradation. While TNKS has previously been shown to be the target of small molecule inhibitors of Wnt/ß-catenin signaling, WIKI4 is structurally distinct from previously identified TNKS inhibitors.