KRAS and BRAF mutations are rare and related to DNA mismatch repair deficiency in gastric cancer from the East and the West: results from a large international multicentre study.

BACKGROUND: Inhibitors of the epidermal growth factor (EGFR) signaling pathway have a major role in the treatment of KRAS wild-type colorectal cancer patients. The EGFR pathway has been shown to be activated in gastric cancer (GC). However, published data on KRAS and BRAF mutation status is limited in GC and has not been compared between GC from different geographic regions. METHODS: The prevalence of KRAS and BRAF mutations was established in 712 GC: 278 GC from the United Kingdom, 230 GC from Japan and 204 GC from Singapore. The relationship between KRAS/BRAF mutation status, DNA mismatch repair (MMR) status, clinicopathological variables and overall survival was analysed. RESULTS: Overall, 30 (4.2%) GC carried a KRAS mutation. In total, 5.8% of the UK GC, 4% of Japan GC and 1.5% of Singapore GC were KRAS mutant. KRAS mutant GC had fewer lymph node metastases in the UK cohort (P=0.005) and were more frequent in elderly patients in the Japan cohort (P=0.034). KRAS mutations were more frequent in MMR-deficient GC in the UK and the Japanese cohort (P<0.05). A BRAF mutation was only detected in a single Japanese GC. CONCLUSIONS: This large multicentre study demonstrated that KRAS mutations and DNA MMR deficiency have a role in a small subgroup of GC irrespective of country of origin, suggesting that this subgroup of GC may have developed along a common pathway. Further studies need to establish whether concomitant mutations or amplifications of other EGFR signalling pathway genes may contribute to the activation of this pathway in GC.

Development of environmental thresholds for streams in agricultural watersheds.

Global increases in consumption of chemical nutrients, application of pesticides, and water withdrawal to enhance agricultural yield have resulted in degraded water quality and reduced water availability. Efforts to safeguard or improve environmental conditions of agroecosystems have usually focused on managing on-farm activities to reduce materials loss and conserve habitat. Another management measure for improving environmental quality is adoption of environmental performance standards (also called outcome-based standards). This special collection of six papers presents the results of four years of research to devise scientifically credible approaches for setting environmental performance standards to protect water quantity and quality in Canadian agriculturally dominated watersheds. The research, conducted as part of Canada's National Agri-Environmental Standards Initiative, aimed to identify Ideal Performance Standards (the desired environmental state needed to maintain ecosystem health) and Achievable Performance Standards (the environmental conditions achievable using currently available and recommended best available processes and technologies). Overviews of the papers, gaps in knowledge, and future research directions are presented. As humans, livestock, and wildlife (both terrestrial and aquatic) experience greater pressures to share the same limited water resources, innovative research is needed that incorporates a landscape perspective, economics, farm practices, and ecology to advance the development and application of tools for protecting water resources in agricultural watersheds.

Effects of the veterinary pharmaceutical salinomycin and its formulation on the plant Brassica rapa.

Veterinary pharmaceuticals are emerging contaminants found throughout the environment, and their presence and effects are a matter of concern. The purpose of this study was to compare the phytotoxicity of salinomycin (pure compound = 96 %) and Sacox 120 (formulated product = 120 g salinomycin/kg) to the plant species Brassica rapa as well as to investigate salinomycin persistence in soil. Calculated EC/IC(50) values for salinomycin and Sacox 120 were 1.10 and 2.88 and 2.19 and 18.03 mg/kg, respectively, based on salinomycin concentration. For exposure of B. rapa to salinomycin, significant adverse effects were observed for growth end points at the greater concentrations. For the reproduction end point (i.e., number of buds), as well as root length and wet mass, significant differences were observed at the lower concentrations (stimulating growth) and adverse effects at the greater concentrations. This study confirmed that the toxic effects of Sacox 120 are attributable to the active ingredient salinomycin. Liquid chromatography-electrospray ionization-mass spectrometry analyses confirmed that exposure concentrations of salinomycin were 90 and 83 % of the nominal concentrations, respectively, in the soils amended with either pure or formulated product. At the end of the experiment, after 14 days, salinomycin concentrations for both tests (salinomycin and Sacox 120) decreased to 6.2 and 5.8 % of the nominal exposure concentrations, respectively. Detected salinomycin concentrations in plant shoots ranged from 3.47 to 41.0 ng/g dry shoot. This study shows the importance of using plants as tools to evaluate environmental risk and as a bridge to relate environment and human health risks.

Application of nitrogen and phosphorus criteria for streams in agricultural landscapes.

Efforts to control eutrophication of water resources in agriculturally dominated ecosystems have focused on managing on-farm activities to reduce nutrient loss; however, another management measure for improving water quality is adoption of environmental performance criteria (or 'outcome-based standards'). Here, we review approaches for setting environmental quality criteria for nutrients, summarize approaches developed in Canada for setting 'ideal' and 'achievable' nutrient criteria for streams in agricultural watersheds, and consider how such criteria could be applied. As part of a 'National Agri-Environmental Standards Initiative', the Government of Canada committed to the development of non-regulatory environmental performance standards that establish total P (TP) and total N (TN) concentrations to protect ecological condition of agricultural streams. Application of four approaches for defining ideal standards using only chemistry data resulted in values for TP and TN spanning a relatively narrow range of concentrations within a given ecoregion. Cross-calibration of these chemically derived standards with information on biological condition resulted in recommendations for TP and TN that would likely protect aquatic life from adverse effects of eutrophication. Non-point source water quality modelling was then conducted in a specific watershed to estimate achievable standards, i.e. chemical conditions that could be attained using currently available and recommended management practices. Our research showed that, taken together, short-term achievable standards and ultimate ideal standards could be used to set policy targets that should, if realized, lower N and P concentrations in Canadian agricultural streams and improve biotic condition.

How does climate variability affect water quality dynamics in Canada's oil sands region?

In Canada's oil sands region, classic boreal hydrology (i.e., winter low flow followed by peaks during spring freshet and then summer flow recession) combined with erosion of both natural and anthropogenically-exposed bitumen results in seasonal and inter-annual variability in stream water chemistry. Using data collected from all seasons over three years (2012-2015), we investigated the mechanisms driving spatial and temporal change in the concentration of 26 water quality parameters for six rivers draining Canada's oil sands region. Mantel tests showed a strong spatial aggregation of climatic drivers (average daily precipitation, accumulated precipitation, snow water equivalent) associated with west versus east discharge patterns. Wavelet analysis highlighted unique watershed attributes, in particular the importance of developed area in lowering responsiveness to seasonal precipitation. Concentrations of most chemical parameters (20 of 23) showed distinct temporal patterns that were correlated with seasonal changes in hydrology which, in turn, were related to changes in weather. Comparison of concentrations observed in this study with those reported in the scientific literature for the same watersheds showed 81% of comparisons differed significantly. This was likely due to the short duration of previous field campaigns and thus the sampling of a very narrow window of the annual streamflow regime.

Catchment properties and the photosynthetic trait composition of freshwater plant communities.

Unlike in land plants, photosynthesis in many aquatic plants relies on bicarbonate in addition to carbon dioxide (CO2) to compensate for the low diffusivity and potential depletion of CO2 in water. Concentrations of bicarbonate and CO2 vary greatly with catchment geology. In this study, we investigate whether there is a link between these concentrations and the frequency of freshwater plants possessing the bicarbonate use trait. We show, globally, that the frequency of plant species with this trait increases with bicarbonate concentration. Regionally, however, the frequency of bicarbonate use is reduced at sites where the CO2 concentration is substantially above the air equilibrium, consistent with this trait being an adaptation to carbon limitation. Future anthropogenic changes of bicarbonate and CO2 concentrations may alter the species compositions of freshwater plant communities.

Vascularisation of Norian CRS bone cement and its replacement by autologous bone when used for orbital reconstruction.

Various bone cements based on calcium phosphate have been used as adjuncts for repairing both the craniofacial and axial skeleton. Ideally these materials should provide initial strength and contour for the reconstruction, and be replaced over time by physiological absorption and bony deposition. Although there is evidence from animal models to support this, opportunities for human studies are rare. Here we offer clinical and histological evidence of this process.

Episodic acidification of 5 rivers in Canada's oil sands during snowmelt: A 25-year record.

Episodic acidification during snowmelt is a natural phenomenon that can be intensified by acidic deposition from heavy industry. In Canada's oil sands region, acid deposition is estimated to be as much as 5% of the Canadian total and large tracks of northeastern Alberta are considered acid-sensitive because of extensive peatland habitats with poorly weathered soils. To identify the frequency, duration and severity of acidification episodes during snowmelt (the predominant hydrological period for delivery of priority pollutants from atmospheric oil sands emissions to surface waters), a 25-year record (1989 to 2014) of automated water quality data (pH, temperature, conductivity) was assembled for 3 rivers along with a shorter record (2012-2014) for another 2 rivers. Acidic episodes (pH<7, ANC<0) were recorded during 39% of all 83 snowmelt events. The severity (duration x magnitude) of episodic acidification increased exponentially over the study period (r2=0.56, P<0.01) and was strongly correlated (P<0.01) with increasing maximum air temperature and weakly correlated with regional land development (P=0.06). Concentrations of aluminum and 11 priority pollutants (Sb, As, Be, Cd, Cr, Cu, Pb, Se, Ag, Tl and Zn) were greatest (P<0.01) during low (<6.5) pH episodes, particularly when coincident with high discharge, such that aluminum and copper concentrations were at times high enough to pose a risk to juvenile rainbow trout (Oncorhynchus mykiss). Although low pH (pH<6.5) was observed during only 8% of 32 acidification episodes, when present, low pH typically lasted 10days. Episodic surface water acidification during snowmelt, and its potential effects on aquatic biota, is therefore an important consideration in the design of long-term monitoring of these typically alkaline (pH=7.72±0.05) rivers.

Tri- and tetrapeptide analogues of kinins as potential renal vasodilators.

Tri- and tetrapeptide analogues were synthesized and evaluated as renal vasodilators. These peptides were prepared by standard coupling reactions, which also provided good yields with hindered alpha-methyl amino acid derivatives. Preliminary evidence of renal vasodilator activity was determined in anesthetized dogs by measuring the effects on renal blood flow and calculating the accompanying changes in renal vascular resistance. The most potent compounds contained, in their basic structure, the L-prolyl-DL-alpha-methylphenylalanyl-L-arginine and L-prolyl-DL-alpha-methylphenylalanylglycyl-L-proline arrays. Substitution on the N-terminal proline with 4-phenylbutyryl and 4-(4-hydroxyphenyl)butyryl side chains produced enhanced renal vasodilator activity and, in certain cases, selectivity for the renal vasculature.

Trisubstituted pyridine leukotriene B4 receptor antagonists: synthesis and structure-activity relationships.

A series of trisubstituted pyridines have been prepared that exhibit in vitro leukotriene B4 (LTB4, 1) receptor antagonist activity. Previous disubstituted pyridines from these labs showed high affinity for the LTB4 receptor but demonstrated agonist activity in functional assays (e.g., 2, Ki = 1 nM). Compound 4, the initial lead compound of this new series, showed only modest affinity by comparison (Ki = 282 nM); however, 4 was a receptor antagonist with no demonstrable agonist activity up to 10 microM. Subsequent modifications of the lipid tail and aryl head group region led to the discovery of aniline 50 (SB 201146). This compound, also free of agonist activity, possesses high affinity for the LTB4 receptor (Ki = 4.7 nM).

Dopaminergic activity of substituted 6-chloro-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepines.

6-Chloro-7,8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepines were synthesized and evaluated as agonists of central and peripheral dopamine receptors. These benzazepines were prepared by cyclization of certain amino alcohols followed by demethylation of the 7,8-dimethoxy groups of the precursors to the 7,8-catecholic moiety. Preliminary evidence of dopaminergic activity was determined in anesthetized dogs by measuring the effects on renal blood flow and calculating the accompanying changes in renal vascular resistance. The most potent compounds contained an hydroxyl group on the 1-phenyl group or were substituted at the 3' position with a chloro, methyl, or trifluoromethyl group. Evidence for central dopaminergic activity was obtained by measuring rotational effects in rats lesioned in the substantia nigra and also in an in vitro assay which measured stimulation of rat striatal adenylate cyclase. The compounds with the best central dopaminergic activity were generally the benzazepines which were the most lipophilic, were substituted on the 3' position of th 1-phenyl group, and contained either a 3-N-methyl or 3-N-allyl group.

(E)-3-[6-[[(2,6-dichlorophenyl)thio]methyl]-3-(2-phenylethoxy)-2- pyridinyl]-2-propenoic acid: a high-affinity leukotriene B4 receptor antagonist with oral antiinflammatory activity.

An extensive structure-activity study based around the high-affinity leukotriene B4 (LTB4) receptor antagonist SB 201146 (1) led to the identification of (E)-3-[6-[[(2,6-dichlorophenyl)-thio]methyl]-3-(2-phenylethoxy)-2- pyridinyl]-2-propenoic acid (3). This compound displays high affinity for the human neutrophil LTB4 receptor (Ki = 0.78 nM), blocks LTB4-induced Ca2+ mobilization with an IC50 of 6.6 +/- 1.5 nM, and demonstrates potent oral and topical antiinflammatory activity in a murine model of dermal inflammation.

(E)-3-[[[[6-(2-carboxyethenyl)-5-[[8-(4- methoxyphenyl)octyl]oxy]-2-pyridinyl]-methyl]thio]methyl]benzoic acid and related compounds: high affinity leukotriene B4 receptor antagonists.

(E)-3-[[[[6-(2-Carboxyethenyl)-5-[[8-(4- methoxyphenyl)octyl]oxy]-2-pyridinyl]methyl]thio]methyl]benzoic acid (11, SB 201993) is a novel, potent LTB4 receptor antagonist. Compound 11 arose from a structure-activity study of a series of trisubstituted pyridines that demonstrated LTB4 receptor antagonist activity. The placement of an additional methylene unit in the sulfur containing chain linking the pyridine and benzoic acid moieties of lead compound 8 (K(i) = 80 nM) resulted in a greater than 10-fold increase in receptor affinity. Additionally, in this new series of compounds, the oxidation state of the sulfur was found to be critical to the activity, i.e., the sulfoxide and sulfone showed substantially lower affinity for the LTB4 receptor. Compound 11 competitively inhibits the binding of [3H]LTB4 to LTB4 receptors on human polymorphonuclear leukocytes with a Ki of 7.1 nM and blocks both the LTB4-induced calcium mobilization and the LTB4-induced degranulation responses in these cells with IC50 values of 131 and 271 nM, respectively. Compound 11 demonstrated oral LTB4 antagonist activity as well as topical antiinflammatory activity in the mouse.

Survival of free DNA encoding antibiotic resistance from transgenic maize and the transformation activity of DNA in ovine saliva, ovine rumen fluid and silage effluent.

To assess the likelihood that the bla gene present in a transgenic maize line may transfer from plant material to the microflora associated with animal feeds, we have examined the survival of free DNA in maize silage effluent, ovine rumen fluid and ovine saliva. Plasmid DNA that had previously been exposed to freshly sampled ovine saliva was capable of transforming competent Escherichia coli cells to ampicillin resistance even after 24 h, implying that DNA released from the diet could provide a source of transforming DNA in the oral cavity of sheep. Although target DNA sequences could be amplified by polymerase chain reaction from plasmid DNA after a 30-min incubation in silage effluent and rumen contents, only short term biological activity, lasting less than 1 min, was observed in these environments, as shown by transformation to antibiotic resistance. These experiments were performed under in vitro conditions; therefore further studies are needed to elucidate the biological significance of free DNA in the rumen and oral cavities of sheep and in silage effluent.

In vitro and in vivo pharmacological characterization of SB 201993, an eicosanoid-like LTB4 receptor antagonist with anti-inflammatory activity.

Leukotriene B4 (LTB4) and 12-(R)-hydroxy-5,8,10,14-eicosatetraenoic acid (12-[R]-HETE) have been postulated to contribute to the pathophysiology of inflammatory diseases. SB 201993, (E)-3-[[[[6-(2-carboxyethenyl)-5-[[8-(4-methoxyphenyl)octyl] oxy]-2-pyridinyl] methyl] thio] methyl] benzoic acid, identified from a chemical series designed as ring-fused analogs of LTB4, was evaluated as an antagonist of LTB4- and 12-(R)-HETE-induced responses in vitro and for anti-inflammatory activity in vivo. SB 201993 competitively antagonized [3-H]-LTB4 binding to intact human neutrophils (Ki = 7.6 nM) and to membranes of RBL 2H3 cells expressing the LTB4 receptor (RBL 2H3-LTB4R; IC50 = 154 nM). This compound demonstrated competitive antagonism of LTB4- and 12-(R)-HETE-induced Ca2+ mobilization responses in human neutrophils (IC50s of 131 nM and 105 nM, respectively) and inhibited LTB4-induced Ca2+ mobilization in human cultured keratinocytes (IC50 = 61 nM), RBL 2H3-LTB4R cells (IC50 = 255 nM) and mouse neutrophils (IC50 = 410 nM). SB 201993 showed weak LTD4-receptor binding affinity (Ki = 1.9 microM) and inhibited 5-lipoxygenase (IC50 of 3.6 microM), both in vitro and ex vivo. In vivo, SB 201993 inhibited LTB4-induced neutrophil infiltration in mouse skin and produced dose-related, long lasting topical anti-inflammatory activity against the fluid and cellular phases of arachidonic acid-induced mouse ear inflammation (ED50 of 580 microg/ear and 390 microg/ear, respectively). Similarly, anti-inflammatory activity was also observed in the murine phorbol ester-induced cutaneous inflammation model (ED50 of 770 and 730 microg/ear, respectively, against the fluid and cellular phases). These results indicate that SB 201993 blocks the actions of LTB4 and 12-(R)-HETE and inhibits a variety of inflammatory responses; and thus may be a useful compound to evaluate the role of these mediators in disease models.

Current Velocity and Its Effect on Aquatic Macrophytes in Flowing Waters.

Current velocity significantly affected the biomass and shoot density of aquatic macrophytes in two slow-flowing rivers in western Canada. Studies of aquatic macrophyte communities at three sites on the Bow River, Alberta, Canada, between 1982 and 1985 showed that biomass decreased with increasing current velocity within the weed bed over the range 0.01-1 m/s; at current speeds in excess of 1 m/s, aquatic macrophytes were rare. Transplant experiments in which Potamogeton pectinatus was grown in pails containing three sediments differing in texture at three sites with different current velocities also demonstrated that biomass and shoot density were affected by both the direct effects of current velocity on plant shoots and its indirect effects on sediment nutrient concentrations. These results indicate that current velocity is an important factor regulating aquatic macrophyte biomass in flowing waters and suggest that even a relatively modest increase in current velocity within weed beds reduces the abundance of submerged aquatic plants.

Comparative study of the ipsilateral full thickness forearm skin graft in closure of radial forearm flap donor site defects.

The early experience of our unit with the technique of ipsilateral full thickness forearm skin grafting of the radial forearm flap donor site defect is described. The technique provides the advantages of a full thickness skin graft whilst avoiding the need to harvest skin from a remote area. We have compared the results with that of split skin grafts used contemporaneously for the same purpose in our unit; the advantages and disadvantages are outlined. Continued success has encouraged us to utilize this technique routinely with certain modifications which we describe.

Management of locally aggressive sinus disease using craniofacial exposure and the galeal frontalis fascia-muscle flap.

Histologically, benign sinus diseases can behave aggressively, potentially causing sight- and life-threatening conditions requiring surgical treatment. Four patients illustrating acute and chronic infection, single and multiple sinus polyposis, and mucoceles are presented and discussed to illustrate how the principles of craniofacial exposure, resection, and reconstruction may be employed to treat aggressive sinus disease. The bicoronal and buccal sulcus approaches minimize visible scarring; wide exposure of the orbital contents and dura reduce the risk of excessive bleeding, neurologic complications, and orbital volume derangements. Immediate cranial bone grafting and the well-vascularized galeal-frontalis myofascial flap obliterate the dead space and isolate the extradural space from contaminated facial cavities, diminishing the risk of infection. The craniofacial surgical approach is particularly appropriate for the management of aggressive frontoethmoidal and multiple sinus involvement because it allows the treatment of sinus cavities under direct vision and complete resection of sinus disease.

Closure of large orocutaneous fistulas in end-stage malignant disease.

Expanding Silastic foam was used to close and dress a large orocutaneous salivary fistula secondary to a massive local recurrence of squamous cell carcinoma. The technique is simple, rapid, and effective. The patient found the dressing superior in every way to the traditional absorbent dressings previously supplied. We recommend this technique for the closure of large defects in the head and neck.

Management of first branchial cleft anomalies: report of two cases.

Embryological anomalies of the first branchial apparatus result in rare forms of developmental abnormality of the head and neck. Their presentation may be similar to other conditions and they may easily be overlooked by the unwary when considering the differential diagnosis of a parotid swelling or a neck sinus. Consequently, they may be mismanaged on one or more occasions. We encountered two patients in whom such problems arose and conclude that appropriate investigation and management by a team skill ed in surgery of this region is necessary if a satisfactory outcome is to be achieved.