Inactivation of p53 Rescues the Maintenance of High Risk HPV DNA Genomes Deficient in Expression of E6

The human papillomavirus DNA genome undergoes three distinct stages of replication: establishment, maintenance and amplification. We show that the HPV16 E6 protein is required for the maintenance of the HPV16 DNA genome as an extrachromosomal, nuclear plasmid in its natural host cell, the human keratinocyte. Based upon mutational analyses, inactivation of p53 by E6, but not necessarily E6-mediated degradation of p53, was found to correlate with the ability of E6 to support maintenance of the HPV16 genome as a nuclear plasmid. Inactivation of p53 with dominant negative p53 rescued the ability of HPV16 E6STOP and E6SAT mutant genomes to replicate as extrachromosomal genomes, though not to the same degree as observed for the HPV16 E6 wild-type (WT) genome. Inactivation of p53 also rescued the ability of HPV18 and HPV31 E6-deficient genomes to be maintained at copy numbers comparable to that of HPV18 and HPV31 E6WT genomes at early passages, though upon further passaging copy numbers for the HPV18 and 31 E6-deficient genomes lessened compared to that of the WT genomes. We conclude that inactivation of p53 is necessary for maintenance of HPV16 and for HPV18 and 31 to replicate at WT copy number, but that additional functions of E6 independent of inactivating p53 must also contribute to the maintenance of these genomes. Together these results suggest that re-activation of p53 may be a possible means for eradicating extrachromosomal HPV16, 18 or 31 genomes in the context of persistent infections.     

Evidence of interpopulation variation in the germination of Eryngium maritimum L. (Apiaceae)

Plant Ecology - Germination is considered as one of the most crucial steps in a plant’s lifetime. The germination of Eryngium maritimum, a dune species located across all European coasts, has...     

Potential cultivation of Hordeum vulgare L. in soils with high mercury background concentrations

Experimental work was carried out under close-to-real conditions to study mercury uptake by Hordeum vulgare L. cultivated in lysimeter experiments. The soil in the lysimeter experiment was obtained from a test plot located near Almadén (Spain) and had a mean mercury content of 22.9 mg kg(-1). A sequence of four crops was sown starting in autumn 2000 and repeated on a yearly basis until 2004. The first crop was grown in the field prior to the extraction of 5 one-cubic-meter lysimeters. The succeeding crops were sown in the lysimeter experiments at the CIEMAT Research Centre (Madrid, Spain). Samples of root and shoot were obtained during the four seasons. Concentrations of mercury at plant maturity in roots vary between I and 3 mg kg(-1) and in straw and grain the concentrations range from 72 to 480 microg kg(-1) and from 5 to 257 microg kg(-1), respectively. In order to assess the potential risk for human health and animal feed, an evaluation of the mercury content in the edible part of the crop has been carried out. According to legislation, there is no human health intoxication risk with a balanced consumption; otherwise, the forage use would have to be controlled.     

Overfishing of small pelagic fishes increases trophic overlap between immature and mature striped dolphins in the Mediterranean Sea

The interactions among diet, ecology, physiology, and biochemistry affect N and C stable isotope signatures in animal tissues. Here, we examined if ecological segregation among animals in relation to sex and age existed by analyzing the signatures of δ(15)N and δ(13)C in the muscle of Western Mediterranean striped dolphins. Moreover, we used a Bayesian mixing model to study diet composition and investigated potential dietary changes over the last two decades in this population. For this, we compared isotope signatures in samples of stranded dolphins obtained during two epizootic events occurring in 1990 and 2007-2008. Mean δ(13)C values for females and males were not significantly different, but age-related variation indicated δ(13)C enrichment in both sexes, suggesting that females and males most likely fed in the same general areas, increasing their consumption of benthic prey with age. Enrichment of δ(15)N was only observed in females, suggesting a preference for larger or higher trophic level prey than males, which could reflect different nutritional requirements. δ(13)C values showed no temporal variation, although the mean δ(15)N signature decreased from 1990 to 2007-2008, which could indicate a dietary shift in the striped dolphin over the last two decades. The results of SIAR indicated that in 1990, hake and sardine together contributed to 60% on the diet of immature striped dolphins, and close to 90% for mature striped dolphins. Conversely, the diet of both groups in 2007-2008 was more diverse, as hake and sardine contributed to less than 40% of the entire diet. These results suggest a dietary change that was possibly related to changes in food availability, which is consistent with the depletion of sardine stocks by fishing.     

Geomicrobiology of Pyrite (FeS2) Dissolution: Case Study at Iron Mountain,California

Geomicrobiology of pyrite weathering at Iron Mountain, CA, was investigated by molecular biological, surface chemical, surface structural, and solution chemical methods in both laboratory and field-based studies. Research focused at sites both within and peripheral to the ore-body. The acid-generating areas we have examined thus far at Iron Mountain (solution pH <1.0, temperature> 35 C) were populated by species other than Thiobacillus ferrooxidans . 16S rDNA bacterial sequence analysis and domain- and specieslevel oligonucleotide probe-based investigations confirmed the presence of planktonic Leptospirillum ferrooxidans and indicated the existence of other species apparently related to other newly described acidophilic chemolithotrophs. T. ferrooxidans was confined to relatively moderate environments (pH 2-3, 20-30 C) that were peripheral to the orebody. Dissolution rate measurements indicated that, per cell, attached and planktonic species contributed comparably in acid release. Surface colonization experiments in the laboratory and field indicated that attachment was specific to sulfides instead of to silicates, occurred in crystallographically preferred orientations, and, after cell division, resulted in a monolayer of cells at a maximum density of 8 X 106 cells cm2. In situ geochemical characterization throughout the year revealed that the microbial community that controlled acid generation varied and could be correlated with seasonal and spatial fluctuations in geochemical conditions.     

Zirconium Nitride for Plasmonic Cloaking of Visible Nanowire Photodetectors

Light scattered by a photodetector disturbs the probing field, resulting in noise. Cloaking is an effective method to reduce this noise. Here we investigate theoretically an emerging plasmonic material, zirconium nitride (ZrN), as a plasmonic cloak for silicon (Si) nanowire-based photodetectors and compare it with a traditional plasmonic material, gold (Au). Using Mie formalism, we have obtained the scattering cancelation across the visible spectrum. We found that ZrN cloaks produce a significant decrease in the scattering from bare Si nanowires, which is 40% greater than that obtained with Au cloaks in the wavelength region of 400–500 nm. The scattering cancelations become comparable at 550 nm, with Au providing a better scattering cancelation compared to ZrN over the wavelength region of 600–700 nm. To include the absorption and provide a measure of overall performance on noise reduction, a figure of merit (FOM), defined as the ratio of the absorption efficiency and the scattering efficiency of the cloaked nanowire to that of the bare Si nanowire, was calculated. We show that the optimized ZrN cloak provides up to 3 times enhancement of the FOM over a bare Si NW and a 60% improvement over an optimized Au-cloaked NW, in the wavelength region of 400–500 nm. An optimized Au-cloaked NW shows up to 17.69 times improvement in the wavelength region of 600–700 nm over a bare Si NW and up to a 2.7 times improvement over an optimized ZrN-cloaked NW. We also predicted the optimal dimensions for the cloaked NWs with respect to the largest FOM at various wavelengths between 400 and 650 nm.

     

Indirect Transmission of Influenza A Virus between Pig Populations under Two Different Biosecurity Settings

Respiratory disease due to influenza virus is common in both human and swine populations around the world with multiple transmission routes capable of transmitting influenza virus, including indirect routes. The objective of this study was to evaluate the role of fomites in influenza A virus (IAV) transmission between pig populations separated by two different biosecurity settings. Thirty-five pigs were divided into four experimental groups: 10 pigs (1 replicate) were assigned to the infected group (I), 10 pigs (2 replicates of 5 pigs) were assigned to the low biosecurity sentinel group (LB), 10 pigs (2 replicates of 5 pigs) were assigned to the medium biosecurity sentinel group (MB), and 5 pigs (1 replicate) were assigned to the negative control group (NC). Eight of 10 pigs in the infected group were inoculated with IAV and 36 hours following inoculation, personnel movement events took place in order to move potentially infectious clothing and personal protective equipment (PPE) to sentinel pig rooms. Following contact with the infected group, personnel moved to the MB group after designated hygiene measures while personnel moved directly to the LB group. Nasal swabs and blood samples were collected from pigs to assess IAV infection status and fomites were sampled and tested via RRT-PCR. All experimentally inoculated pigs were infected with IAV and 11 of the 144 fomite samples collected following contact with infected pigs were low level positive for IAV genome. One replicate of each sentinel groups LB and MB became infected with IAV and all five pigs were infected over time. This study provides evidence that fomites can serve as an IAV transmission route from infected to sentinel pigs and highlights the need to focus on indirect routes as well as direct routes of transmission for IAV.     

Differential Organ Distribution,Pathogenicity and Benomyl Sensitivity of Colletotrichum spp. from Blackberry Plants in Northern Colombia

Blackberry anthracnose, caused by Colletotrichum spp., is an important disease of cultivated blackberry in the world. In Colombia, it is the number one limiting factor for commercial production. This study was conducted to determine the species of Colletotrichum infecting blackberry plants as well as the organ distribution, pathogenicity and response to benomyl of the isolated strains. Sixty isolates from stems (n = 20), thorns (n = 20) and inflorescences (n = 20) were identified as Colletotrichum acutatum and Colletotrichum gloeosporioides by a species‐specific polymerase chain reaction (PCR). Both Colletotrichum species were found in the same plant but on different organs. Colletotrichum gloeosporioides species predominated in thorn lesions (n = 16) and C. acutatum in stems (n = 15) and inflorescence (n = 15). Pathogenicity assays on detached blackberry organs demonstrated differences between the two species with an average period of lesion development of 8.7 days for C. gloeosporioides and 10.3 days for C. acutatum. Wound inoculated organs had 90% disease development compared to 17.5% in non‐wounded. All C. acutatum isolates (n = 34) were benomyl tolerant, whereas C. gloeosporioides isolates (n = 26) were 30.7% sensitive and 69.2% moderately tolerant. Phylogenetic analysis with ITS sequences of a subset of 18 strains showed that strains classified as C. gloeosporioides had 100% identity to Colletotrichum kahawae, which belongs to the C. gloeosporioides species complex, whereas C. acutatum strains clustered into two different groups, with high similarity to the A2 and the A4 molecular groups. These data demonstrate for the first time the differential distribution of both species complexes in blackberry plant organs and further clarifies the taxonomy of the strains.     

Isolation and characterization of thylakoid membranes from the filamentous cyanobacterium Nostoc punctiforme

Nostoc   punctiforme strain Pasteur Culture Collection (PCC) 73102, a sequenced filamentous cyanobacterium capable of nitrogen fixation, is used as a model organism for characterization of bioenergetic processes during nitrogen fixation in Nostoc . A protocol for isolating thylakoid membranes was developed to examine the biochemical and biophysical aspects of photosynthetic electron transfer. Thylakoids were isolated from filaments of N.   punctiforme by pneumatic pressure-drop lysis. The activity of photosynthetic enzymes in the isolated thylakoids was analysed by measuring oxygen evolution activity, fluorescence spectroscopy and electron paramagnetic resonance spectroscopy. Electron transfer was found functional in both PSII and PSI. Electron transfer measurements in PSII, using diphenylcarbazide as electron donor and 2,6-dichlorophenolindophenol as electron acceptor, showed that 80% of the PSII centres were active in water oxidation in the final membrane preparation. Analysis of the membrane protein complexes was made by 2D gel electrophoresis, and identification of representative proteins was made by mass spectrometry. The ATP synthase, several oligomers of PSI, PSII and the NAD(P)H dehydrogenase (NDH)-1L and NDH-1M complexes, were all found in the gels. Some differences were noted compared with previous results from Synechocystis sp. PCC 6803. Two oligomers of PSII were found, monomeric and dimeric forms, but no CP43-less complexes. Both dimeric and monomeric forms of Cyt b ₆/ f could be observed. In all, 28 different proteins were identified, of which 25 are transmembrane proteins or membrane associated ones.