Ecophysiological response of aspen (Populus tremuloides) and jack pine (Pinus banksiana) to atmospheric nitrogen deposition on reconstructed boreal forest soils in the Athabasca oil sands region.

Oil and gas extraction in the Athabasca Oil Sands Region of northeastern Alberta, Canada has increased anthropogenic nitrous oxide (NOx) and ammonia (NH3) emissions over the past three decades, leading to a potential increase in N deposition. Deposition on reclaimed sites was hypothesized to be higher than in surrounding boreal forests, but had not been quantified. The objective of this study was to assess the implications of this potentially increased deposition on reclaimed aspen (Populus tremuloides Michx.) and pine (Pinus banksiana Lamb.) ecosystems through the use of several N status indicators, including N deposition, total and available concentrations in plants and soils, and δ15N values in deposition and plants and soils. Atmospheric N deposition, which was dominated by ammonium (NH4+), averaged 24 kg N ha-1 year-1 as bulk precipitation and 6 kg N ha-1 year-1 as throughfall. Increased N deposition influenced the N cycle in both aspen and pine stands. Aspen appeared to be actively biocycling N as indicated by a closed N cycle, resulting in minimal N losses. Whereas the N cycle in pine may be more open as indicated by the dominance of soil nitrate (NO3-), and enrichment of 15N in available soil NH4+, root and foliar N. Therefore, we suggest that pine stands on reclamation sites may be at kinetic N saturation where the rate of N inputs exceeds vegetation and soil N net sinks, and do not require additional N fertilizer inputs.

Ellagitannins as synergists of ACV on the replication of ACV-resistant strains of HSV 1 and 2.

The plant-derived polyphenolic compounds castalagin, vescalagin and grandinin (C-glucosidic ellagitannins containing nonahydroxyterphenoyl) manifested a strong inhibitory effect on the replication of acyclovir-resistant strains of herpes simplex viruses (HSV) type 1 and 2 in MDBK cells in focus forming units (i.e., microscopically registered microplaques) reduction assay and in two variants of cytopathic effect inhibition test. The effect on the acyclovir (ACV)-resistant herpes simplex virus type 1 (HSV-1) strain was markedly higher compared to that on the ACV-resistant herpes simplex virus type 2 (HSV-2). The three compounds showed comparable levels of antiviral activity against ACV-resistant HSV strains, in contrast with previous results where castalagin exerted the highest degree of activity against wild type HSV strains (Vilhelmova et al., 2011). Combinations of ellagitannins and ACV were tested on the ACV-resistant strains of both HSV-1 and 2 and produced synergistic effects that were revealed by applying the three-dimensional approach of Prichard and Shipman (1990). The ellagitannin(s)-ACV combination applied against ACV-resistant HSV-1 produced a much stronger synergistic effect compared to the effect observed against ACV-resistant HSV-2. The study of the effects of the combination ellagitannin(s)-ACF on intact cell monolayers did not show any toxicity resulting from interaction between the two substances. Altogether, the results obtained in this study demonstrate the highly promising potential of these plant polyphenols as antiherpetic agents.

Trastuzumab induced in vivo tissue remodelling associated in vitro with inhibition of the active forms of AKT and PTEN and RhoB induction in an ovarian carcinoma model.

BACKGROUND: The incidence of ovarian cancer has been increasing worldwide and it is currently the leading cause of death from gynaecological malignancy. Unlike breast cancer, the prognostic role of the human epidermal growth factor receptor-2 (HER-2) in ovarian carcinoma remains controversial. METHODS: The aim of this preclinical study was to further characterise the biological, molecular and cellular effects of trastuzumab (Herceptin) using NIH-OVCAR-3 and derived cell lines both in vitro and in vivo. RESULTS: In vitro assessments have shown that trastuzumab treatment inhibited total and phosphorylated HER-2. This was associated with inhibition of the phosphorylated form of phosphatase and tensin homologue (PTEN), mitogen-activated protein kinase and AKT, and the total level of p27(kip). Inhibition of PTEN is associated with phosphorylated MEK1/2 upregulation, suggesting a specific inhibition of the protein phosphatase function of PTEN. Moreover, trastuzumab induced the upregulation of RhoB. These molecular modifications promote inhibition of cell migration and potentially restoration of tumour cell contact inhibition. RhoB induction in NIH-OVCAR-3 control cell lines mimics the molecular and cellular trastuzumab long-time exposition effects. RhoB inhibition in NIH-OVCAR-3 long-time exposed to trastuzumab cell line reverses the cellular and molecular effects observed in this model. In vivo examinations have shown that these changes are also associated with the restoration of structural, morphological and normal functions of the peritoneum of an ovarian carcinoma mouse model. CONCLUSION: These results provide an indication of the mechanisms underlying the anti-tumour activity of trastuzumab that strongly implicate RhoB in an ovarian carcinoma model that does not show HER-2 amplification or overexpression. These findings highlight that trastuzumab effects involve a possible cross-talk between RhoB and PTEN in the early stages of tumour re-growth in a model of micrometastatic ovarian cancer.

Recreating a functioning forest soil in reclaimed oil sands in northern alberta: an approach for measuring success in ecological restoration.

During oil-sands mining all vegetation, soil, overburden, and oil sand is removed, leaving pits several kilometers wide and up to 100 m deep. These pits are reclaimed through a variety of treatments using subsoil or a mixed peat-mineral soil cap. Using nonmetric multidimensional scaling and cluster analysis of measurements of ecosystem function, reclamation treatments of several age classes were compared with a range of natural forest ecotypes to discover which treatments had created ecosystems similar to natural forest ecotypes and at what age this occurred. Ecosystem function was estimated from bioavailable nutrients, plant community composition, litter decomposition rate, and development of a surface organic layer. On the reclamation treatments, availability of nitrate, calcium, magnesium, and sulfur were generally higher than in the natural forest ecotypes, while ammonium, P, K, and Mn were generally lower. Reclamation treatments tended to have more bare ground, grasses, and forbs but less moss, lichen, shrubs, trees, or woody debris than natural forests. Rates of litter decomposition were lower on all reclamation treatments. Development of an organic layer appeared to be facilitated by the presence of shrubs. With repeated applications of fertilizers, measured variables for the peat-mineral amendments fell within the range of natural variability at about 20 yr. An intermediate subsoil layer reduced the need for fertilizer and conditions resembling natural forests were reached about 15 yr after a single fertilizer application. Treatments over tailings sand receiving only one application of fertilizer appeared to be on a different trajectory to a novel ecosystem.

Nitrogen mineralization and microbial activity in oil sands reclaimed boreal forest soils.

Organic materials including a peat-mineral mix (PM), a forest floor-mineral mix (L/S), and a combination of the two (L/PM) were used to cap mineral soil materials at surface mine reclamation sites in the Athabasca oil sands region of northeastern Alberta, Canada. The objective of this study was to test whether LFH provided an advantage over peat by stimulating microbial activity and providing more available nitrogen for plant growth. Net nitrification, ammonification, and N mineralization rates were estimated from field incubations using buried bags. In situ gross nitrification and ammonification rates were determined using the 15N isotope pool dilution technique, and microbial biomass C (MBC) and N (MBN) were measured by the chloroform fumigation-extraction method. All reclaimed sites had lower MBC and MBN, and lower net ammonification and net mineralization rates than a natural forest site (NLFH) used as a control, but the reclamation treatment using LFH material by itself had higher gross and net nitrification rates. A positive correlation between in situ moisture content, dissolved organic N, MBC, and MBN was observed, which led us to conduct a moisture manipulation experiment in the laboratory. With the exception of the MBN for the L/S treatment, none of the reclamation treatments ever reached the levels of the natural site during this experiment. However, materials from reclamation treatments that incorporated LFH showed higher respiration rates, MBC, and MBN than the PM treatment, indicating that the addition of LFH as an organic amendment may stimulate microbial activity as compared to the use of peat alone.

Recognition characters in peptide-polyphenol complex formation.

Dietary polyphenols have received attention for their anti-oxidative, anti-carcinogenic and anti-neurodegenerative effects. Polyphenols bind to proteins leading to the formation of soluble or insoluble protein-polyphenol complexes which could significantly influence their biological activities. NMR and molecular modeling studies were performed to investigate the influence of the bulk, flexibility and hydrophobicity of polyphenols on the association with bradykinin, the peptide model. Our results show that the strength of the interactions could be positively correlated with polyphenol hydrophobicity and a comparison between pentagalloylglucose and vescalagin indicated that flexibility might play a positive role in the interaction with peptides and proteins.

Identification of the lipophilic factor produced by macrophages that stimulates steroidogenesis.

Macrophages are known to release a lipophilic factor that stimulates testosterone production by Leydig cells. This macrophage-derived factor (MDF) is thought to be physiologically relevant, because removal of macrophages from the testis results in altered testosterone secretion and reduced fertility. The purpose of the present study was to purify this factor, elucidate its chemical structure, and determine whether it is both present in the testis and acts when injected intratesticularly. Culture media from testicular and peritoneal macrophages were extracted with ether, and the organic phase was sequentially purified on C18, silica, and cyano-HPLC columns. MDF was detected using a rat Leydig cell bioassay, with testosterone secretion being the end point. Purified material and crude ether extracts were analyzed by gas chromatography/mass spectrometry and nuclear magnetic resonance spectroscopy. The time of elution of MDF from both testicular and peritoneal macrophages was identical on all three HPLC columns. A single peak was observed when MDF, obtained from the final HPLC column, was analyzed by gas chromatography. The MS fragmentation pattern of purified material from both peritoneal and testicular macrophages was identical to that of a reference preparation of 25-hydroxycholesterol. Also, the nuclear magnetic resonance spectrum of MDF was similar to that of authentic 25-hydroxycholesterol. When 25-hydroxycholesterol was subjected to the identical purification scheme as MDF, it was found to elute at the same times as MDF on all three columns and elicited activity in the Leydig cell bioassay as expected. Control medium purified identically did not contain 25-hydroxycholesterol or have biological activity. Ether extracts of testis contained 25-hydroxycholesterol, indicating that this compound is present under physiological conditions. Similarly, when 25-hydroxycholesterol was injected into the testis of adult rats, testosterone production was increased within 3 h. Taken together, these data indicate that the lipophilic factor produced by macrophages that stimulates steroidogenesis is 25-hydroxycholesterol.