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Determination of growth-phase dependent influences exerted by prions on yeast lipid content using HPTLC—densitometry

Bui Q., Sherma J., Fried B., Hines K.

Acta Chromatographica

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2016

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Vol. 28, no. 3

373--385

EN

Prions of the baker’s yeast Saccharomyces cerevisiae allow for the inheritance of complex traits based solely on the acquisition of cytoplasmic protein aggregates and confer distinctive phenotypes to the cells which harbor them, creating heterogeneity within an otherwise clonal cell population. These phenotypes typically arise from a loss-of-function of the prion-forming protein that is unable to perform its normal cellular function( s) while sequestered in prion amyloid aggregates, but the specific biochemical consequences of prion infection are poorly understood. To begin to address this issue, we initiated a direct investigation into the potential control that yeast prions exert over fungal lipid content by utilizing the prions [URE3] and [PSI+], the first two prions discovered in yeast. We utilized silica gel high-performance thin-layer chromatography (HPTLC)—densitometry to conduct pair-wise quantifications of the relative levels of free sterols, free fatty acids, and triacylglycerols [petroleum ether—diethyl ether—acetic acid (80:20:1) mobile phase, phosphomolybdic acid (PMA) detection reagent]; steryl esters and squalene (hexane—petroleum ether—diethyl ether—acetic acid (50:20;5:1), PMA]; and phosphatidylethanolamine, phosphatidylcholine, and phosphatidylinositol (chloroform– diethyl ether—acetic acid (65:25:4.5), cupric sulfate—phosphoric acid) in otherwise clonal prion-infected ([PSI+] or [URE3]) and prion-free ([psi−] or [ure-o]) cells in two growth phases: log-phase and stationary phase. Our analysis revealed multiple statistically significant differences (p < 0.00625) between prion-infected and prion-free cells. Interestingly, prion-induced changes varied dramatically by growth phase, indicating that prions exert differential influences on cell physiology between log and stationary growth. Further experimental replication and extension of the analysis to other prions is expected to resolve additional physiological effects of prion infection. This investigation demonstrates that HPTLC—densitometry is an effective method for studying prion-induced alterations in lipid content in yeast.

2

Charakterystyka peptydów i białek zaangażowanych w utrzymywanie homeostazy jonów miedzi w układzie nerwowym człowieka

Komoszyńska A., Maćkiewicz Z.

Wiadomości Chemiczne

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2007

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[Z] 61, 7-8

547-565

EN

Copper is an essential metal in living organisms. They all require copper as a catalytic cofactor for biological processes such as respiration, iron transport, oxidative stress protection and pigmentation. Nevertheless, exposure to the excess of copper can damage cells and organs. Copper ions and complexes can induce formation of reactive oxygen species, that can damage biomolecules including unsaturated lipids and DNA. In the past few years exciting advances have been made toward understanding how copper is transported within cells. Dysfunction of copper metabolism can lead to its excess or defi-ciency. Two examples of illness related to alterations in copper metabolism are Menkes and Wilson diseases. The understanding of copper homeostasis has become important in clinical medicine as the metal could be involved in the pathogenesis of some neurological disorders such as Alzheimer, Parkinson or prion diseases.

3

Notatki chaotyczne. Cz. 50. O prionach

Siemion I. Z.

Wiadomości Chemiczne

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2004

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[Z] 58, 1-2

131--141

4

Priony i choroby prionowe

Laboratorium - Przegląd Ogólnopolski

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2004

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nr 5

48--50

PL

Spożyte wraz z pokarmem priony przedostają się do układu pokarmowego, a następnie do śledziony, kłębków Peyera, migdałków i grudek chłonnych, gdzie ulegają replikacji. Powielone priony są następnie przenoszone przez komórki limfoidalne do tkanki glejowej i nerwowej.

5

Priony - zagrożenia biologiczne spuścizną byłego tysiąclecia

Zużewicz M.A.

Bezpieczeństwo Pracy : nauka i praktyka

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2001

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nr 6

11-13

PL

W pracy omówiono problemy dotyczące nowego rodzaju zagrożeń biologicznych. Przedstawiono historię badań niezwykłego czynnika biologicznego jakim jest prion. Zwrócono uwagę na występowanie tego patogenu w środowisku człowieka i jego działanie na makroogranizmy. Wskazano grupy zawodowe, które mogą być eksponowane na działanie prionów. Przedstawiono dane i hipotezy, przydatne w kreowaniu bezpieczniejszego miejsca pracy oraz kierunki postępowania w celu zmniejszenia ryzyka zachorowania.

EN

The paper discusses a new type of biohazards. The author presents the history of research on the prion, an unusual biological factor. She points to the presence Of this pathogen in the human environment and its influence on macroorganisms, and lists occupational groups possibly exposed to prions. Data and hypotheses useful in creating a less harmful workplace and ways of minimizing the risk of falling ill are presented.

6

Sterylizacja odpadów zakażonych prionami

Rathberger K.

LAB Laboratoria, Aparatura, Badania

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1998

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R. 3, nr 2

18-19