Boundary spanning at the science-policy interface: the practitioners' perspectives.

Cultivating a more dynamic relationship between science and policy is essential for responding to complex social challenges such as sustainability. One approach to doing so is to "span the boundaries" between science and decision making and create a more comprehensive and inclusive knowledge exchange process. The exact definition and role of boundary spanning, however, can be nebulous. Indeed, boundary spanning often gets conflated and confused with other approaches to connecting science and policy, such as science communication, applied science, and advocacy, which can hinder progress in the field of boundary spanning. To help overcome this, in this perspective, we present the outcomes from a recent workshop of boundary-spanning practitioners gathered to (1) articulate a definition of what it means to work at this interface ("boundary spanning") and the types of activities it encompasses; (2) present a value proposition of these efforts to build better relationships between science and policy; and (3) identify opportunities to more effectively mainstream boundary-spanning activities. Drawing on our collective experiences, we suggest that boundary spanning has the potential to increase the efficiency by which useful research is produced, foster the capacity to absorb new evidence and perspectives into sustainability decision-making, enhance research relevance for societal challenges, and open new policy windows. We provide examples from our work that illustrate this potential. By offering these propositions for the value of boundary spanning, we hope to encourage a more robust discussion of how to achieve evidence-informed decision-making for sustainability.

Design and testing of miniaturized plasma sensor for measuring hypervelocity impact plasmas.

An increasingly notable component of the space environment pertains to the impact of meteoroids and orbital debris on spacecraft and the resulting mechanical and electrical damages. Traveling at speeds of tens of km/s, when these particles, collectively referred to as hypervelocity particles, impact a satellite, they vaporize, ionize, and produce a radially expanding plasma that can generate electrically harmful radio frequency emission or serve as a trigger for electrostatic discharge. In order to measure the flux, composition, energy distribution, and temperature of ions and electrons in this plasma, a miniaturized plasma sensor has been developed for carrying out in-situ measurements in space. The sensor comprises an array of electrostatic analyzer wells split into 16 different channels, catering to different species and energy ranges in the plasma. We present results from numerical simulation based optimization of sensor geometry. A novel approach of fabricating the sensor using printed circuit boards is implemented. We also describe the test setup used for calibrating the sensor and show results demonstrating the energy band pass characteristics of the sensor. In addition to the hypervelocity impact plasmas, the plasma sensor developed can also be used to carry out measurements of ionospheric plasma, diagnostics of plasma propulsion systems, and in other space physics experiments.

Beyond single-marker analyses: mining whole genome scans for insights into treatment responses in severe sepsis.

Management of severe sepsis, an acute illness with high morbidity and mortality, suffers from the lack of effective biomarkers and largely empirical predictions of disease progression and therapeutic responses. We conducted a genome-wide association study using a large randomized clinical trial cohort to discover genetic biomarkers of response to therapy and prognosis utilizing novel approaches, including combination markers, to overcome limitations of single-marker analyses. Sepsis prognostic models were dominated by clinical variables with genetic markers less informative. In contrast, evidence for gene-gene interactions were identified for sepsis treatment responses with genetic biomarkers dominating models for predicting therapeutic responses, yielding candidates for replication in other cohorts.

Designing pharmacogenetic projects in industry: practical design perspectives from the Industry Pharmacogenomics Working Group.

Pharmacogenetic association studies have the potential to identify variations in DNA sequence which impact drug response. Identifying these DNA variants can help to explain interindividual variability in drug response; this is the first step in personalizing dosing and treatment regimes to a patient's needs. There are many intricacies in the design and analysis of pharmacogenetic association studies, including having adequate power, selecting proper endpoints, detecting and correcting the effects of population stratification, modeling genetic and nongenetic covariates accurately, and validating the results. At this point there are no formal guidelines on the design and analysis of pharmacogenetic studies. The Industry Pharmacogenomics Working Group has initiated discussions regarding potential guidelines for pharmacogenetic study design and analyses (http://i-pwg.org) and the results from these discussions are presented in this paper.

Genomewide association for schizophrenia in the CATIE study: results of stage 1.

Little is known for certain about the genetics of schizophrenia. The advent of genomewide association has been widely anticipated as a promising means to identify reproducible DNA sequence variation associated with this important and debilitating disorder. A total of 738 cases with DSM-IV schizophrenia (all participants in the CATIE study) and 733 group-matched controls were genotyped for 492,900 single-nucleotide polymorphisms (SNPs) using the Affymetrix 500K two-chip genotyping platform plus a custom 164K fill-in chip. Following multiple quality control steps for both subjects and SNPs, logistic regression analyses were used to assess the evidence for association of all SNPs with schizophrenia. We identified a number of promising SNPs for follow-up studies, although no SNP or multimarker combination of SNPs achieved genomewide statistical significance. Although a few signals coincided with genomic regions previously implicated in schizophrenia, chance could not be excluded. These data do not provide evidence for the involvement of any genomic region with schizophrenia detectable with moderate sample size. However, a planned genomewide association study for response phenotypes and inclusion of individual phenotype and genotype data from this study in meta-analyses hold promise for eventual identification of susceptibility and protective variants.

Common polymorphisms of CYP2C19 and CYP2C9 affect the pharmacokinetic and pharmacodynamic response to clopidogrel but not prasugrel.

BACKGROUND: Thienopyridines are metabolized to active metabolites that irreversibly inhibit the platelet P2Y(12) adenosine diphosphate receptor. The pharmacodynamic response to clopidogrel is more variable than the response to prasugrel, but the reasons for variation in response to clopidogrel are not well characterized. OBJECTIVE: To determine the relationship between genetic variation in cytochrome P450 (CYP) isoenzymes and the pharmacokinetic/pharmacodynamic response to prasugrel and clopidogrel. METHODS: Genotyping was performed for CYP1A2, CYP2B6, CYP2C19, CYP2C9, CYP3A4 and CYP3A5 on samples from healthy subjects participating in studies evaluating pharmacokinetic and pharmacodynamic responses to prasugrel (60 mg, n = 71) or clopidogrel (300 mg, n = 74). RESULTS: In subjects receiving clopidogrel, the presence of the CYP2C19*2 loss of function variant was significantly associated with lower exposure to clopidogrel active metabolite, as measured by the area under the concentration curve (AUC(0-24); P = 0.004) and maximal plasma concentration (C(max); P = 0.020), lower inhibition of platelet aggregation at 4 h (P = 0.003) and poor-responder status (P = 0.030). Similarly, CYP2C9 loss of function variants were significantly associated with lower AUC(0-24) (P = 0.043), lower C(max) (P = 0.006), lower IPA (P = 0.046) and poor-responder status (P = 0.024). For prasugrel, there was no relationship observed between CYP2C19 or CYP2C9 loss of function genotypes and exposure to the active metabolite of prasugrel or pharmacodynamic response. CONCLUSIONS: The common loss of function polymorphisms of CYP2C19 and CYP2C9 are associated with decreased exposure to the active metabolite of clopidogrel but not prasugrel. Decreased exposure to its active metabolite is associated with a diminished pharmacodynamic response to clopidogrel.

Steroid-free maintenance immunosuppression after heart transplantation.

BACKGROUND: Steroids are routinely used in almost all immunosuppressive protocols after cardiac transplantation. The metabolic side effects of steroids are well known and could lead to significant morbidity and mortality in the posttransplant period. There is growing evidence to suggest that steroids may not be a requirement for adequate immunosuppression and that morbidity may be reduced by withdrawing steroids in select patients. We have reviewed our series of patients undergoing heart transplantation in whom steroids were weaned postoperatively. METHODS: We retrospectively reviewed all adult patients undergoing heart transplantation at our institution between November 1993 and April 2000 treated with a-triple-drug immunosuppressive regimen. Medications were recorded at discharge and at 6, 12, and 24 months posttransplant to determine the success of steroid weaning. Freedom from infection and rejection as well as overall survival was calculated using Kaplan-Meier methods. RESULTS: By 24 months posttransplant, almost 70% of patients were receiving double-drug therapy. Survival for the entire group was excellent with 1-, 3-, and 5-year survival of 98%+/-2.0%, 93.2%+/-3.8%, and 88.3%+/-6.0%, respectively. Freedom from rejection at 6 months was 60.7%+/-6.5%, at 1 year was 60.7%+/-6.5%, and at 2 years was 58.5%+/-6.7%. Infectious complications were low with freedom from infection at 6 months of 78.5+/-5.5%, at 1 year of 76.5%+/-5.7%, and at 2 years of 72.0%+/-6.2%. CONCLUSIONS: Our data suggest that an immunosuppressive regimen without long-term steroid administration results in excellent survival rates without an apparent increase in rejection or infectious complications.

Measuring postural changes in blood pressure in the healthy elderly.

BACKGROUND: Background The reproducibility of postural changes in blood pressure of a healthy elderly population determined using standard clinical measurements is not known. OBJECTIVE: To assess the differences in reproducibility of postural changes in blood pressure in healthy elderly subjects 1 and 3 min after standing within a day and between visits spaced 6 weeks apart. METHODS: Casual readings of blood pressures of supine and standing subjects were measured twice during the day by the same observer on two occasions 6 weeks apart using a semi-automatic syphgmomanometer. Twenty-two subjects with no known risk factors for orthostatic hypotension (13 men) aged 69+/-3 years (mean+/-SD) with a mean initial screening supine blood pressure of 153+/-19/88+/-11 mmHg were recruited. RESULTS: There were significant differences(P<0.001) between the postural changes both for systolic and for diastolic blood pressure between 1 and 3 min of standing, the largest falls occurring after 1 min of standing, though we found no variation between morning and afternoon measurements and between visits. The coefficients of reproducibility between visits for the postural changes in blood pressure after 1 and 3 min of standing were large both for systolic and for diastolic blood pressure, ranging from 9.8 to 29.3 mmHg. CONCLUSIONS: There was a consistent and significant difference between the postural changes in blood pressure after 1 and 3 min of standing for this healthy elderly population, but there was no significant difference between the postural changes in morning and afternoon measurements and between visits. This marked variability in the postural change in blood pressure with duration of standing must be taken into account when assessing the prevalence of orthostatic hypotension and the effects of treatment in patients with orthostatic falls in blood pressure.

Audit of opioid prescribing: the effect of hospital guidelines.

The effects of prescribing guidelines for analgesia were assessed by auditing prescriptions for opioids before and after the introduction of hospital prescribing guidelines. Opioid prescriptions were collected by the pharmacy department over a 2-week period in November 1994 and repeated in November 1995. Following the initial audit, analgesic prescribing guidelines were introduced. A statistically significant increase was achieved in the number of prescriptions that were correct for both dose and frequency according to both the British National Formulary recommendations (40-61%; p < 0.001) and our Acute Pain Service guidelines (16-26%; p < 0.05). There was a statistically significant decrease in the number of prescriptions that were inadequate for both dose and frequency according to both the British National Formulary recommendations (18-3%; p < 0.001) and our Acute Pain Service guidelines (36-17%; p = 0.001). The use of accessible prescribing guidelines promotes demonstrable improvements in opioid prescribing.

Comparison of diurnal systolic blood pressure change as defined by wrist actigraphy, fixed time periods and cusum.

AIMS: To assess whether differences exist in nocturnal blood pressure (BP) levels and the diurnal BP change when using fixed time and wrist actigraphy methods to define the night-time period. METHODS: Untreated hypertensive (n = 48) and normotensive (n = 33) subjects (mean age 67 years: range 29-90) underwent simultaneous 24-h ambulatory BP monitoring and wrist actigraph monitoring. The diurnal BP change and nocturnal BP levels were assessed using two fixed night-time definitions-a reference period of 22.00-06.59 and also 00.00-05.59, as well as unedited and edited actigraph values and cumulative sums (cusums) analysis. RESULTS: The reference definition of night-time 22.00-06.59 resulted in the highest values for night-time BP compared to other definitions (p < 0.05), although actigraph defined night-time BP values were not significantly different from the more restrictive definition of night-time (00.00-05.59). Restrictive night-time, edited and unedited actigraph and cusums values for the day-night difference were all significantly greater than the value calculated using the reference night-time period. Dipping status changed significantly depending on which definition of night-time was used. CONCLUSIONS: Significant differences exist in nocturnal BP levels and circadian change between the various methods for defining night-time. The routine use of wrist actigraphy does not however appear to alter the value of night-time BP levels when compared to a more restrictive fixed-time definition of night-time.

What method should be used to define 'night' when assessing diurnal systolic blood pressure variation in the elderly?

Diurnal blood pressure (BP) variation can be assessed by cusums-derived measures and by the day-night BP difference from time-, activity- and diary-defined 'night' periods. Reproducibility of diurnal systolic BP (SBP) variation by these different methods was studied in 19 active elderly normotensives, mean age 68.5 years. Subjects underwent simultaneous 24 h BP (Spacelabs 90207) and activity (Gaehwiler wrist actigraph) monitoring on two occasions (median interval 70 days). On the first occasion, mean diurnal SBP variation was 15.1 +/- 8.1 mm Hg by fixed-time definition of 'night' (22.07). When compared with 22-07 defined 'night' period, actigraph- and diary-defined 'night-time' was significantly reduced (-60 +/- 49, and -48 +/- 51 min, respectively) and consequently diurnal SBP variation was significantly greater at 18.2 +/- 8.1 mm Hg and 17.6 +/- 8.4 mm Hg, respectively. Actigraph recordings were also used to exclude 'night' BP readings associated with activity, but this did not significantly alter the diurnal SBP variation. Cusums-derived circadian alteration magnitude resulted in the greatest value for SBP variation (23.4 +/- 6.7 mm Hg). However, reproducibility of diurnal SBP variation was poor by fixed-time method with a coefficient of variation (CV) of > 50%, and only improved to 40% with diary use. Actigraph measurements, even if used to exclude BP values associated with disturbed sleep, did not improve this further. Cusums-derived measures of diurnal variation slightly improved reproducibility with a CV of 34.6% and may be a better method in the assessment of diurnal BP variation in the elderly.

Genomic position affects the expression of tobacco mosaic virus movement and coat protein genes.

Alterations in the genomic position of the tobacco mosaic virus (TMV) genes encoding the 30-kDa cell-to-cell movement protein or the coat protein greatly affected their expression. Higher production of 30-kDa protein was correlated with increased proximity of the gene to the viral 3' terminus. A mutant placing the 30-kDa open reading frame 207 nucleotides nearer the 3' terminus produced at least 4 times the wild-type TMV 30-kDa protein level, while a mutant placing the 30-kDa open reading frame 470 nucleotides closer to the 3' terminus produced at least 8 times the wild-type TMV 30-kDa protein level. Increases in 30-kDa protein production were not correlated with the subgenomic mRNA promoter (SGP) controlling the 30-kDa gene, since mutants with either the native 30-kDa SGP or the coat protein SGP in front of the 30-kDa gene produced similar levels of 30-kDa protein. Lack of coat protein did not affect 30-kDa protein expression, since a mutant with the coat protein start codon removed did not produce increased amounts of 30-kDa protein. Effects of gene positioning on coat protein expression were examined by using a mutant containing two different tandemly positioned tobamovirus (TMV and Odontoglossum ringspot virus) coat protein genes. Only coat protein expressed from the gene positioned nearest the 3' viral terminus was detected. Analysis of 30-kDa and coat protein subgenomic mRNAs revealed no proportional increase in the levels of mRNA relative to the observed levels of 30-kDa and coat proteins. This suggests that a translational mechanism is primarily responsible for the observed effect of genomic position on expression of 30-kDa movement and coat protein genes.

Dopamine D1 and D2 receptor interactions in the MPTP-treated marmoset.

In a modified MPTP model of Parkinson's disease in the marmoset, both L-DOPA and the dopamine D2 agonist quinpirole were found to exhibit anti-bradykinetic activity. Both the dopamine D1 agonist SKF38393 and the D1 antagonist SCH23390 reduced the anti-bradykinetic action of L-DOPA and quinpirole. These results are discussed with respect to partial agonist activity of SKF38393 and the possibility that other dopamine receptors may be required for anti-Parkinsonian drug activity.

A gene near the plasmid pSa origin of replication encodes a nuclease.

We have cloned and sequenced a gene (nuc) from the IncW plasmid pSa which shows amino acid sequence similarity to staphylococcal nuclease (EC 3.1.4.7) and to the parB locus of plasmid RP4. The 525 bp open reading frame encodes a 174-amino-acid potential polypeptide of 19.7 kDa. Expression of the gene was confirmed using an in vitro transcription-translation assay which produced a protein of identical size. Nuclease activity was demonstrated using DNA as the substrate in toluidine blue-DNA agar plates. The deduced amino acid sequence revealed a signal sequence, and TnphoA insertion within the open reading frame indicated that a portion of the protein is transported across the bacterial cell membrane.

The osa gene of pSa encodes a 21.1-kilodalton protein that suppresses Agrobacterium tumefaciens oncogenicity.

The incompatibility group W plasmid pSa suppresses Agrobacterium tumefaciens oncogenicity (J. Loper and C. Kado, J. Bacteriol. 139:591-596, 1979). The oncogenic suppressive activity was localized to a 3.1-kb region of pSa by Tn5 mutagenesis and deletion analysis. Within this fragment, a 1.1-kb subclone bearing oncogenic suppressive activity was subjected to further characterization. Nucleotide sequencing of the 1.1-kb fragment revealed a 570-bp open reading frame (ORF1) that has a coding capacity for a protein of 21.1 kDa. Sequencing of flanking regions revealed a second ORF (ORF2) located 3 bp upstream of ORF1, with a coding capacity for a protein of 22.8 kDa. Gene fusions of these ORFs to a T7 phi 10 expression system in Escherichia coli resulted in the synthesis of polypeptides of the predicted sizes. An E. coli promoter consensus sequence was not found in the expected positions in the region preceding ORF1. However, several sequences with similarity to the consensus -10 sequence of the A. tumefaciens vir gene promoters were found upstream of ORF1. Potential translational start signals are upstream of ORF1 and ORF2. These sequences showed no significant similarity at the nucleotide or amino acid levels with those in available data bases. However, the C-terminal portion of the ORF1 protein is rich in hydrophobic residues. Perhaps oncogenicity suppression is effected by an association of this protein with the Agrobacterium membrane such that T-DNA transfer is blocked.

Procedure for assessing the behavioral effects of novel anti-parkinsonian drugs in normal and MPTP-treated marmosets following central microinfusions.

Comparison between drug effects in MPTP- and non-MPTP-treated marmosets following intracerebroventricular (ICV) and intraputamen administration are given in the present report. Both side-effect profiles and the ability to reverse MPTP-induced hypokinesia were assessed using a variety of dopamine receptor agonists (quinpirole, PHNO, ADTN), cholinergic antagonists (scopolamine, secoverine, himbacine), a glutamate receptor antagonist (MK801) and a 5-HT receptor agonist (8-OH-DPAT). Our results show that direct infusion of agents into the putamen or via the ICV route can reverse parkinsonian-like symptoms in marmosets. Furthermore, chronic implantation of cannulae into the putamen or ventricles of nonhuman primates can be useful in assessing the therapeutic activity of agents that do not readily cross the blood-brain barrier.

Behavioural effects of serotonin agonists and antagonists in the rat and marmoset.

The present study was conducted to investigate the effects of various 5-hydroxytryptamine (5-HT) agonists and antagonists on motor behaviour in rats and marmosets. Various motor-based responses were assessed after central or peripheral administration of 5-HT agents to rats and marmosets. Drugs acting as agonists at the 5-HT1A receptor (8-OHDPAT, gepirone, BMY-7378, NAN-190, PAPP (LY165163) and flesinoxan) and 5-HT2/1C receptors (DOI) were found to reverse neuroleptic-induced catalepsy in the rat whereas 5-HT2/1C antagonists (mianserin, ritanserin and ICI-170,809) and the 5-HT1 antagonist ((+/-)pindolol) increased catalepsy. Agonists acting at 5-HT3 receptors (phenylbiguanide and 2-methyl-5-HT) had no effect on catalepsy. The putative 5-HT1A antagonist, (+/-) pindolol, attenuated the reversal of catalepsy by 8-OHDPAT. Although both 8-OHDPAT and BMY-7378 were tested, only the latter was found to reduce apomorphine-induced stereotypy. Bilateral or unilateral infusions of 8-OHDPAT, BMY-7378 or pindolol into the substantia nigra of non-lesioned rats had no effect on spontaneous locomotor or rotational activity, respectively. However, 8-OHDPAT and BMY-7378 were found to increase or decrease motor activity, after injection into the median or dorsal raphe nuclei, respectively. Finally, 8-OHDPAT and BMY-7378 were found to be inactive against MPTP-induced bradykinesia in the marmoset. It is concluded that both 5-HT1A and 5-HT2/1C receptors are involved in the anti-cataleptic effects of 5-HT agents. The 5-HT1A receptors are probably situated within the raphe, whereas the location of the 5-HT2/1C receptors remains undetermined.

Neuroleptic-induced catalepsy as a model of Parkinson's disease. I. Effect of dopaminergic agents.

Catalepsy was observed in the rat following intrastriatal injections of the dopamine antagonists sulpiride or fluphenazine and after subcutaneous administration of fluphenazine. The neuroleptic-induced catalepsy was reversed by the classical anti-parkinsonian agent L-DOPA and by agents that function through dopamine systems such as d- and methamphetamine and the direct D2 receptor agonist quinpirole. The D1 agonist SKF 38393, and the D1/D2 agonist apomorphine, were ineffective in this model. These results support limited use of the rat catalepsy model for the screening of potential anti-parkinsonian compounds and indicate that this procedure can provide valuable information concerning striatal dopamine function.

Neuroleptic-induced catalepsy as a model of Parkinson's disease. II. Effect of glutamate antagonists.

Subcutaneous administration of fluphenazine elicits catelepsy that can be attenuated by the glutamate antagonists MK801 and phencyclidine (PCP). 3-[-(+)-2-carboxy piperazine-4-yl]-propyl-1-phosphanate (CPP) was found to be ineffective in this model. Intrastriatal injections of sulpiride or fluphenazine were also found to induce catalepsy which could be attenuated by MK801 and PCP. These results illustrate that nondopaminergic compounds might possibly be of value in the treatment of Parkinson's disease. Furthermore it was demonstrated that this paradigm can be utilized to investigate neurotransmitter interactions within the striatum. This was clearly emphasized by the observation that bilateral administration of MK801 into the striatum increased basal locomotor activity.