Polyamine Synthesis and Interconversion by the Microsporidian Encephalitozoon cuniculi

Polyamines are small cationic molecules necessary for growth and differentiation in all cells. Although mammalian cells have been studied extensively, particularly as targets of polyamine antagonists, i.e. antitumor agents, polyamine metabolism has also been studied as a potential drug target in microorganisms. Since little is known concerning polyamine metabolism in the microsporidia, we investigated it in Encephalitozoon cuniculi, a microspordian associated with disseminated infections in humans. Organisms were grown in RK-13 cells and harvested using Percoll gradients. Electron microscopy indicated that the fractions banding at 1.051-1.059/g/ml in a microgradient procedure, and 1.102-1.119/g/ml in a scaled-up procedure were nearly homogenous, consisting of pre-emergent (immature) spores which showed large arrays of ribosomes near polar filament coils. Intact purified pre-emergent spores incubated with [1H] ornithine and methionine synthesized putrescine, spermidine, and spermine, while [14C]spermine was converted to spermidine and putrescine. Polyamine production from ornithine was inhibitable by DL-alpha-difluoromethylornithine (DFMO) but not by DL-alpha-difluoromethylarginine (DFMA). Cell-free extracts from mature spores released into the growth media had ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (AdoMetdc), and spermidine/spermine N1-acetyltransferase (SSAT) activities. ODC activity was inhibited by DFMO, but not by DFMA. AdoMetdc was putrescine-stimulated and inhibited by methylglyoxal-bis(guanylhydrazone); arginine decarboxylase activity could not be detected. It is apparent from these studies that Encephalitozoon cuniculi pre-emergent spores have a eukaryotic-type polyamine biosynthetic pathway and can interconvert exogenous polyamines. Pre-emergent spores were metabolically active with respect to polyamine synthesis and interconversion, while intact mature spores harvested from culture supernatants had little metabolic activity.     

A new species of Clivia (Amaryllidaceae) endemic to the Pondoland Centre of Endemism, South Africa

Clivia robusta B.G. Murray, Ran, de Lange, Hammett, Truter et Swanevelder sp. nov. (Amaryllidaceae) is a tubular, pendulous-flowered Clivia species, restricted to the Pondoland Centre of Endemism, South Africa. The unique morphology, distribution, karyotype and molecular fingerprint distinguish it from all other pendulous-flowered species in the genus.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 369–374.     

The Development of Activities of Some Enzymes Concerned with Glycollate Metabolism in Greening Bean Leaves

The activities of phosphoglycollate phosphatase (EC 3.1.3.18 [EC] ),glycollate oxidase (EC 1.1.3.1 [EC] .). catalase (EC 1.11.1.6 [EC] ), theperoxisomal NADH-glyoxylate reductase (EC 1.1.1.26 [EC] ) which isconsidered to function as a hydroxypyruvate reductase in theperoxisomes, and the chloro-plastic NADPH-dependent glyoxylatereductaae, have been measured in extracts prepared from 14-d-olddark-grown bean leaves during the course of their greening inresponse to exposure to continuous illumination. All of theenzymes were found in the dark-grown leaves and on a per-leafbasis the activities increased from 6- to 12-fold with the exceptionof a 2–3-fold increase of NADPH-dependent glyoxylate reductaseduring 96-h greening, while the activities either remained constantor declined during similar periods in darkness. Initial lagperiods were evident before the illumination-induced increasesin enzyme activities. As D-threo-chloramphenicol did not affectthe increase in activity of any of these enzymes it would appearthat the increases were in no way dependent on protein synthesisby 70S ribosomes, or on the development of photosynthetic activity.     

A COMPARISON OF 30° AND 37° AS INCUBATION TEMPERATURES IN THE PRESUMPTIVE COLIAEROGENES TEST FOR RAW AND PASTEURIZED MILK

SUMMARY: Incubation at 30° and 37° for the presumptive coli-aerogenes test for raw and pasterurized milk has been investigated. There were more positives at the lower temperature and it is suggested that in this test, incubation at 30° might provide a much better guide to the hygienic quality of both raw and pasterurized milk. The ability of the coli-aerogenes bacteria studied to ferment lactose in MacConkey's broth at 30° but not at 37° was found to be a stable factor which was unchanged by prolonged storage on agar slopes at room temperature or on continued incubation in MacConkey's broth at temperatures above the optimum for lactose fermentation.     

Characterization of the TRBP domain required for Dicer interaction and function in RNA interference

Background  

Dicer, Ago2 and TRBP are the minimum components of the human RNA-induced silencing complex (RISC). While Dicer and Ago2 are RNases, TRBP is the double-stranded RNA binding protein (dsRBP) that loads small interfering RNA into the RISC. TRBP binds directly to Dicer through its C-terminal domain.     

The cellular prion protein and its role in Alzheimer disease

The cellular prion protein (PrP^C) is a membrane-bound glycoprotein especially abundant in the central nervous system (CNS). The scrapie prion protein (PrP^Sc, also termed prions) is responsible of transmissible spongiform encephalopathies (TSE), a group of neurodegenerative diseases which affect humans and other mammal species, although the presence of PrP^C is needed for the establishment and further evolution of prions.The present work compares the expression and localization of PrP^C between healthy human brains and those suffering from Alzheimer disease (AD).In both situations we have observed a rostrocaudal decrease in the amount of PrP^C within the CNS, both by immunoblotting and immunohistochemistry techniques. PrP^C is higher expressed in our control brains than in AD cases. There was a neuronal loss and astogliosis in our AD cases. There was a tendency of a lesser expression of PrP^C in AD cases than in healthy ones. And in AD cases, the intensity of the expression of the unglycosylated band is higher than the di- and monoglycosylated bands.With regards to amyloid plaques, those present in AD cases were positively labeled for PrP^C, a result which is further supported by the presence of PrP^C in the amyloid plaques of a transgenic line of mice mimicking AD.The work was done according to Helsinki Declaration of 1975, and approved by the Ethics Committee of the Faculty of Medicine of the University of Navarre.Key words: cellular prion protein, Alzheimer disease, transgenic mice     

Avifauna response to hurricanes: regional changes in community similarity

Global climate models predict increases in the frequency and intensity of extreme climatic events such as hurricanes, which may abruptly alter ecological processes in forests and thus affect avian diversity. Developing appropriate conservation measures necessitates identifying patterns of avifauna response to hurricanes. We sought to answer two questions: (1) does avian diversity, measured as community similarity, abundance, and species richness, change in areas affected by hurricane compared with unaffected areas, and (2) what factors are associated with the change(s) in avian diversity? We used North American Breeding Bird Survey data, hurricane track information, and a time series of Landsat images in a repeated measures framework to answer these questions. Our results show a decrease in community similarity in the first posthurricane breeding season for all species as a group, and for species that nest in the midstory and canopy. We also found significant effects of hurricanes on abundance for species that breed in urban and woodland habitats, but not on the richness of any guild. In total, hurricanes produced regional changes in community similarity largely without significant loss of richness or overall avian abundance. We identified several potential mechanisms for these changes in avian diversity, including hurricane‐induced changes in forest habitat and the use of refugia by birds displaced from hurricane‐damaged forests. The prospect of increasing frequency and intensity of hurricanes is not likely to invoke a conservation crisis for birds provided we maintain sufficient forest habitat so that avifauna can respond to hurricanes by shifting to areas of suitable habitat.