Design and Syntheses of Fluorescent Cytosine Analogues

The avid hybridization of peptide nucleic acid (PNA) to DNA and RNA along with the molecule’s biological stability has led it to be used in both antisense and antigene capacities. PNA acts against translation via a steric blockade mechanism. It is therefore reasonable to assume that increased heteroduplex stability could lead to increased potency. Two ways of doing this were explored. I) N-Terminal attachment of a platinous chloride chelating moiety to PNA complementary to Xenopus noggin was synthesized with the objective of selective, covalent platination of the target transcript in vivo. Phenotypes consistent with knockdown of the selected gene product were observed; however, it remains unclear whether this is solely due to site-specific platination. II) The use of the modified cytosine nucleobase [bis-o-(aminoethoxy)phenyl]pyrrolocytosine (boPhpC) was previously shown to increase heteroduplex stability, putatively by interaction with guanine at both the Watson-Crick and Hoogsteen faces. PNAs containing this base were synthesized to target mutant huntingtin mRNA - of which the product is the causative agent of Huntington’s disease – and tested in patient derived fibroblasts where selective inhibition of mutant huntintin was observed with concomitant fluorescence imaging. Modified nucleobases find use in fields ranging from materials science to cytogenetics and has been an area of much endeavour over the past years. Modifications at the 5-position of uracil abound but examples of similarly modified cytosine are lacking. Rapidly developing the inventory of cytosine analogues is a primary goal of ours. We aim to synthesize compounds that may be used as base discriminating fluorophores, created pre- or post-synthetically. Through the reaction of 5-ethynyldeoxycytidine with 1,3-dipoles such as nitrile oxides or azides a series of heterocycle-appended cytosine analogues have been prepared and their fluorescence properties studied. They exhibit moderate to good quantum yields and with high sensitivity to their environment and are considered good candidates for further use as base-discriminating fluorophores. Based on the known pyrrolocytosine scaffold, Indole-3-acetamide substituted deoxypyrrolocytidine (IAMpC) has shown the highest degree of solvatochromism for any pyrrolocytosine analogue known to date. The synthesis of oligodeoxynucleotides containing this base was carried out; however, final deprotection of the base does not proceed smoothly and the modification was found to be slightly destabilizing towards duplex formation

Protocol for Monitoring Fish Communities in Small Streams in the Heartland Inventory and Monitoring Network, Version 2.0

Executive Summary Fish communities are an important component of aquatic systems and are good bioindicators of ecosystem health. Land use changes in the Midwest have caused sedimentation, erosion, and nutrient loading that degrades and fragments habitat and impairs water quality. Because most small wadeable streams in the Heartland Inventory and Monitoring Network (HTLN) have a relatively small area of their watersheds located within park boundaries, these streams are at risk of degradation due to adjacent land use practices and other anthropogenic disturbances. Shifts in the physical and chemical properties of aquatic systems have a dramatic effect on the biotic community. The federally endangered Topeka shiner (Notropis topeka) and other native fishes have declined in population size due to habitat degradation and fragmentation in Midwest streams. By protecting portions of streams on publicly owned lands, national parks may offer refuges for threatened or endangered species and species of conservation concern, as well as other native species. This protocol describes the background, history, justification, methodology, data analysis and data management for long-term fish community monitoring of wadeable streams within nine HTLN parks: Effigy Mounds National Monument (EFMO), George Washington Carver National Monument (GWCA), Herbert Hoover National Historic Site (HEHO), Homestead National Monument of America (HOME), Hot Springs National Park (HOSP), Pea Ridge National Military Park (PERI), Pipestone National Monument (PIPE), Tallgrass Prairie National Preserve (TAPR), and Wilson\u27s Creek National Battlefield (WICR). The objectives of this protocol are to determine the status and long-term trends in fish richness, diversity, abundance, and community composition in small wadeable streams within these nine parks and correlate the long-term community data to overall water quality and habitat condition (DeBacker et al. 2005)

Characterising the impact of sex on severe asthma (SA) in the UK Severe Asthma Registry (UKSAR)

Peer reviewedPostprin

FOLFIRINOX for advanced pancreatic cancer:The Princess Margaret Cancer Centre experience

BACKGROUND: FOLFIRINOX has been shown to significantly increase both overall survival (OS) and progression-free survival (PFS) in metastatic pancreas cancer. There is limited data regarding the treatment of locally advanced pancreatic cancer. We present a retrospective study of patients with both locally advanced and metastatic pancreas cancer using FOLFIRINOX as first-line therapy in our centre. METHODS: This is a retrospective review of patients treated with FOLFIRINOX for pancreatic cancer at Princess Margaret Cancer Centre, between December 2011 and July 2014. The primary objective was to evaluate the efficacy and safety of FOLFIRINOX when used with dose modifications. RESULTS: One hundred two patients were identified; 66 metastatic and 36 locally advanced. Sixty-eight per cent of patients initiated treatment with a dose reduction. The median (95% CI) OS in the metastatic group was 13.1 (6.3–16.1) months with full dose and 12.9 (10.3–30.1) months with modified dose. The median (95% CI) OS in the locally advanced group was 11.1 (6.1–not reached) months with full dose and 23 (not reached–not reached) months with modified dose. The median (95% CI) PFS in the metastatic group was 6.2 (4.9–15.2) months with full dose and 8.7 (5.7–12.9) months with modified dose. The median (95% CI) PFS in the locally advanced group was 11.1 (3.1–not reached) months with full dose and 10.4 (6.8–not reached) months with modified dose. Grade 3/4 haematologic adverse events were observed in 43% of patients. Grade 3/4 non-haematologic adverse events were observed in 28% of patients. Patient well-being significantly improved from baseline to cycle 4 (P=0.002). CONCLUSIONS: Efficacy was achievable with dose-modified FOLFIRINOX in daily setting. The safety of FOLFIRINOX remains a concern with a high rate of grades 3 and 4 neutropaenia despite dose reduction

Resilience of ecosystem service delivery in grasslands in response to single and compound extreme weather events

Extreme weather events are increasing in frequency and magnitude with profound effects on ecosystem functioning. Further, there is now a greater likelihood that multiple extreme events are occurring within a single year. Here we investigated the effect of a single drought, flood or compound (flood + drought) extreme event on temperate grassland ecosystem processes in a field experiment. To assess system resistance and resilience, we studied changes in a wide range of above- and below-ground indicators (plant diversity and productivity, greenhouse gas emissions, soil chemical, physical and biological metrics) during the 8 week stress events and then for 2 years post-stress. We hypothesized that agricultural grasslands would have different degrees of resistance and resilience to flood and drought stress. We also investigated two alternative hypotheses that the combined flood + drought treatment would either, (A) promote ecosystem resilience through more rapid recovery of soil moisture conditions or (B) exacerbate the impact of the single flood or drought event. Our results showed that flooding had a much greater effect than drought on ecosystem processes and that the grassland was more resistant and resilient to drought than to flood. The immediate impact of flooding on all indicators was negative, especially for those related to production, and climate and water regulation. Flooding stress caused pronounced and persistent shifts in soil microbial and plant communities with large implications for nutrient cycling and long-term ecosystem function. The compound flood + drought treatment failed to show a more severe impact than the single extreme events. Rather, there was an indication of quicker recovery of soil and microbial parameters suggesting greater resilience in line with hypothesis (A). This study clearly reveals that contrasting extreme weather events differentially affect grassland ecosystem function but that concurrent events of a contrasting nature may promote ecosystem resilience to future stress

Probing the Role of Melanocortin Type 1 Receptor Agonists in Diverse Immunological Diseases

Background: The melanocortin α-melanocyte stimulating hormone (α-MSH), an endogenous peptide with high affinity for the melanocortin 1 receptor (MC1r), has demonstrated prevention and reversal of intestinal and ocular inflammation in animal models. Preclinical studies were performed to determine whether two MC1r receptor agonists, PL-8177 and PL-8331, exhibit actions and efficacy similar to α-MSH in preventing and reversing intestinal and ocular inflammation.Methods: Both PL-8177 and PL-8331 were assessed in a Eurofins LeadProfilingScreen selectivity panel including 72 in vitro assays. PL-8177 and PL-8331 were evaluated in an in vitro assay using human whole blood stimulated by lipopolysaccharide to determine inhibition of tumor necrosis factor alpha (TNF-α); for comparison, adrenocorticotropic hormone (ACTH) and α-MSH were used as positive controls. PL-8177, dosed at 0.5, 1.5, and 5.0 μg, was assessed in a cannulated rat model of dinitrobenzene sulfonic acid (DNBS)-induced bowel inflammation versus vehicle and oral sulfasalazine. PL-8177 was also dosed at 0.3 mg/kg/mouse injected intraperitoneally versus untreated controls and α-MSH treatment in mice with experimental autoimmune uveitis (EAU). PL-8331 at 3 doses, 3 times daily, was evaluated in a murine model of scopolamine-induced dry eye disease (SiccaSystemTM model), versus twice-daily Restasis® and Xiidra®.Results: Both PL-8177 and PL-8331 demonstrated no significant activity at the 1 μm concentration in any of the 72 in vitro assays. PL-8177 and PL-8331 inhibited lipopolysaccharide-induced TNF-α to a similar degree as ACTH and α-MSH. In the DNBS rat model of bowel inflammation, PL-8177 was significantly superior to untreated controls at all 3 doses (P < 0.05) in reducing bowel inflammation parameters, with effects similar to sulfasalazine. In the murine EAU model, PL-8177 significantly reduced retinal inflammation scores versus untreated controls (P = 0.0001) over 3–5 weeks, and to a similar degree as α-MSH. In the murine scopolamine-induced model of dry eye disease, PL-8331 reduced corneal fluorescein staining scores at all doses, significantly (P = 0.02) for the highest dose (1 × 10-5 mg⋅mL-1), and similarly to Restasis®; Xiidra® demonstrated no effect.Conclusion: The MC1r receptor agonists PL-8177 and PL-8331 exhibited actions similar to those of α-MSH in preventing and reversing intestinal and ocular inflammation in preclinical disease models

Strategies to optimize the impact of nutritional surveys and epidemiological studies

The development of nutrition and health guidelines and policies requires reliable scientific information. Unfortunately, theoretical considerations and empirical evidence indicate that a large percentage of science-based claims rely on studies that fail to replicate. The session "Strategies to Optimize the Impact of Nutrition Surveys and Epidemiological Studies" focused on the elements of design, interpretation, and communication of nutritional surveys and epidemiological studies to enhance and encourage the production of reliable, objective evidence for use in developing dietary guidance for the public. The speakers called for more transparency of research, raw data, consistent data-staging techniques, and improved data analysis. New approaches to collecting data are urgently needed to increase the credibility and utility of findings from nutrition epidemiological studies. Such studies are critical for furthering our knowledge and understanding of the effects of diet on health

Protocol for Monitoring Aquatic Invertebrates of Small Streams in the Heartland Inventory & Monitoring Network, Version 2.1

Executive Summary The Heartland Inventory and Monitoring Network (HTLN) is a component of the National Park Service’s (NPS) strategy to improve park management through greater reliance on scientific information. The purposes of this program are to design and implement long-term ecological monitoring and provide information for park managers to evaluate the integrity of park ecosystems and better understand ecosystem processes. Concerns over declining surface water quality have led to the development of various monitoring approaches to assess stream water quality. Freshwater streams in network parks are threatened by numerous stressors, most of which originate outside park boundaries. Stream condition and ecosystem health are dependent on processes occurring in the entire watershed as well as riparian and floodplain areas; therefore, they cannot be manipulated independently of this interrelationship. Land use activities—such as timber management, landfills, grazing, confined animal feeding operations, urbanization, stream channelization, removal of riparian vegetation and gravel, and mineral and metals mining—threaten stream quality. Accordingly, the framework for this aquatic monitoring is directed towards maintaining the ecological integrity of the streams in those parks. Invertebrates are an important tool for understanding and detecting changes in ecosystem integrity, and they can be used to reflect cumulative impacts that cannot otherwise be detected through traditional water quality monitoring. The broad diversity of invertebrate species occurring in aquatic systems similarly demonstrates a broad range of responses to different environmental stressors. Benthic invertebrates are sensitive to the wide variety of impacts that influence Ozark streams. Benthic invertebrate community structure can be quantified to reflect stream integrity in several ways, including the absence of pollution sensitive taxa, dominance by a particular taxon combined with low overall taxa richness, or appreciable shifts in community composition relative to reference condition. Furthermore, changes in the diversity and community structure of benthic invertebrates are relatively simple to communicate to resource managers and the public. To assess the natural and anthropogenic processes influencing invertebrate communities, this protocol has been designed to incorporate the spatial relationship of benthic invertebrates with their local habitat including substrate size and embeddedness, and water quality parameters (temperature, dissolved oxygen, pH, specific conductance, and turbidity). Rigid quality control and quality assurance are used to ensure maximum data integrity. Detailed standard operating procedures (SOPs) and supporting information are associated with this protocol

Assessing mesoscale material response under shock & isentropic compression via high-resolution line-imaging VISAR.

Of special promise for providing dynamic mesoscale response data is the line-imaging VISAR, an instrument for providing spatially resolved velocity histories in dynamic experiments. We have prepared two line-imaging VISAR systems capable of spatial resolution in the 10-20 micron range, at the Z and STAR facilities. We have applied this instrument to selected experiments on a compressed gas gun, chosen to provide initial data for several problems of interest, including: (1) pore-collapse in copper (two variations: 70 micron diameter hole in single-crystal copper) and (2) response of a welded joint in dissimilar materials (Ta, Nb) to ramp loading relative to that of a compression joint. The instrument is capable of resolving details such as the volume and collapse history of a collapsing isolated pore