Identification of differentially expressed genes in a Giardia lamblia WB C6 clone resistant to nitazoxanide and metronidazole.

OBJECTIVES The characterization of differential gene expression in Giardia lamblia WB C6 strain C4 resistant to metronidazole and nitazoxanide using microarray technology and quantitative real-time PCR. METHODS In a previous study, we created and characterized the G. lamblia WB C6 clone C4 resistant to nitazoxanide and metronidazole. In this study, using a microarray-based approach, we have identified open-reading frames (ORFs) that were differentially expressed in C4 when compared with its wild-type WB C6. Using quantitative real-time PCR, we have validated the expression patterns of some of those ORFs, focusing on chaperones such as heat-shock proteins in wild-type and C4 trophozoites. In order to induce an antigenic shift, trophozoites of both strains were subjected to a cycle of en- and excystation. Expression of selected genes and resistance to nitazoxanide and metronidazole were investigated after this cycle. RESULTS Forty of a total of 9115 ORFs were found to be up-regulated and 46 to be down-regulated in C4 when compared with wild-type. After a cycle of en- and excystation, resistance of C4 to nitazoxanide and metronidazole was lost. Resistance formation and en-/excystation were correlated with changes in expression of ORFs encoding for major surface antigens such as the variant surface protein TSA417 or AS7 ('antigenic shift'). Moreover, expression patterns of the cytosolic heat-shock protein HSP70 B2, HSP40, and of the previously identified nitazoxanide-binding proteins nitroreductase and protein disulphide isomerase PDI4 were correlated with resistance and loss of resistance after en-/excystation. C4 trophozoites had a higher thermotolerance level than wild-type trophozoites. After en-/excystation, this tolerance was lost. CONCLUSIONS These results suggest that resistance formation in Giardia to nitazoxanide and metronidazole is correlated with altered expression of genes involved in stress response such as heat-shock proteins.

Regional vegetation changes in the tropical Andes expressed as arboreal pollen (AP%), standardised ordination scores (DCA z-scores), and aquatic vegetation ratios (D/SS); files used for plots

We compare eight pollen records reflecting climatic and environmental change from the tropical Andes. Our analysis focuses on the last 50 ka, with particular emphasis on the Pleistocene to Holocene transition. We explore ecological grouping and downcore ordination results as two approaches for extracting environmental variability from pollen records. We also use the records of aquatic and shoreline vegetation as markers for lake level fluctuations, and precipitation change. Our analysis focuses on the signature of millennial-scale variability in the tropical Andes, in particular, Heinrich stadials and Greenland interstadials. We identify rapid responses of the tropical vegetation to this climate variability, and relate differences between sites to moisture sources and site sensitivity.

(Table 1) Main constituents of coccolith size fractions expressed as carbonate contribution at Bass River during the PETM

Coccoliths, calcite plates produced by the marine phytoplankton coccolithophores, have previously shown a large array of carbon and oxygen stable isotope fractionations (termed "vital effects"), correlated to cell size and hypothesized to reflect the varying importance of active carbon acquisition strategies. Culture studies show a reduced range of vital effects between large and small coccolithophores under high CO2, consistent with previous observations of a smaller range of interspecific vital effects in Paleocene coccoliths. We present new fossil data examining coccolithophore vital effects over three key Cenozoic intervals reflecting changing climate and atmospheric partial pressure of CO2 (pCO2). Oxygen and carbon stable isotopes of size-separated coccolith fractions dominated by different species from well preserved Paleocene-Eocene thermal maximum (PETM, ~56 Ma) samples show reduced interspecific differences within the greenhouse boundary conditions of the PETM. Conversely, isotope data from the Plio-Pleistocene transition (PPT; 3.5-2 Ma) and the last glacial maximum (LGM; ~22 ka) show persistent vital effects of ~2 per mil. PPT and LGM data show a clear positive trend between coccolith (cell) size and isotopic enrichment in coccolith carbonate, as seen in laboratory cultures. On geological timescales, the degree of expression of vital effects in coccoliths appears to be insensitive topCO2 changes over the range ~350 ppm (Pliocene) to ~180 ppm (LGM). The modern array of coccolith vital effects arose after the PETM but before the late Pliocene and may reflect the operation of more diverse carbon acquisition strategies in coccolithophores in response to decreasing Cenozoic pCO2.

The contract of affreightment as expressed in charterparties and bills of lading /

Includes index.

Territorial slavery question : observations on Senator Douglas' views of popular sovereignty, as expressed in Harper's Magazine for September, 1859.

Printed in double columns.

The booke of the Universall kirk of Scotland: wherein the headis and conclusionis devysit be the ministers and commissionaris of the particular kirks thereof, are especially expressed and contained;

Different edition published under title: Acts and proceedings of the General assemblies of the Kirk of Scotland, from the year M.D. LX.

Opinions concerning standardized milk expressed by dairy leaders in agricultural colleges.

Mode of access: Internet.

The opinions of Sir Robert Peel, expressed in Parliament and in public.

Mode of access: Internet.

The bloody buoy, thrown out as a warning to the political pilots of America; or, A faithful relation of a multitude of acts of horrid barbarity, such as the eye never witnessed, the tongue never expressed, or the imagination conceived, until the commencement of the French revolution.

Mode of access: Internet.

An inquiry into the Scriptural doctrine concerning the devil and Satan : and into the extent of duration expressed by the terms olim, aion, and aionios, rendered everlasting, forever, etc. in the common version, and especially when applied to punishment /

Later eds. have title: Three inquiries ...

An inquiry into the Scriptural doctrine concerning the devil and Satan : and into the extent of duration expressed by the terms olim, aion, and aionios, rendered everlasting, forever, etc. in the common version, and especially when applied to punishment /

Later eds. have title: Three inquiries ...