Implementing post-normal science with or for EU policy actors: using quantitative story-telling.

There is increasing recognition of the wicked nature of the intertwined climate, biodiversity and economic crises, and the need for adaptive, multi-scale approaches to understanding the complexity of both the problems and potential responses. Most science underpinning policy responses to sustainability issues, however, remains overtly apolitical and focussed on technical innovation; at odds with a critical body of literatures insisting on the recognition of systemic problem framing when supporting policy processes. This paper documents the experience of implementing a mixed method approach called quantitative story-telling (QST) to policy analysis that explicitly recognises this normative dimension, as the methodology is part of a post-normal science (PNS) toolkit. The authors reflect on what was learnt when considering how QST fared as a tool for science-policy interaction, working with European Union (EU) level policy actors interested in sustainable agriculture and sustainable development goal 2. These goals-also known as UN Agenda 2030-are the latest institutionalisation of the pursuit of sustainable development and the EU has positioned itself as taking a lead in its implementation. Thus, the paper illustrates our experience of using PNS as an approach to science policy interfaces in a strategic policy context; and illustrates how the challenges identified in the science-policy literature are amplified when working across multiple policy domains and taking a complex systems approach. Our discussion on lessons learnt may be of interest to researchers seeking to work with policy-makers on complex sustainability issues.

Chromatin structure of two genomic sites for targeted transgene integration in induced pluripotent stem cells and hematopoietic stem cells.

Achieving transgene integration into preselected genomic sites is currently one of the central tasks in stem cell gene therapy. A strategy to mediate such targeted integration involves site-specific endonucleases. Two genomic sites within the MBS85 and chemokine (C-C motif) receptor 5 (CCR5) genes (AAVS1 and CCR5 zinc-finger nuclease (CCR5-ZFN) sites, respectively) have recently been suggested as potential target regions for integration as their disruption has no functional consequence. We hypothesized that efficient transgene integration maybe affected by DNA accessibility of endonucleases and therefore studied the transcriptional and chromatin status of the AAVS1 and CCR5 sites in eight human induced pluripotent stem (iPS) cell lines and pooled CD34+ hematopoietic stem cells (HSCs). Matrix chromatin immunoprecipitation (ChIP) assays demonstrated that the CCR5 site and surrounding regions possessed a predominantly closed chromatin configuration consistent with its transcriptional inactivity in these cell types. In contrast, the AAVS1 site was located within a transcriptionally active region and exhibited an open chromatin configuration in both iPS cells and HSCs. To show that the AAVS1 site is readily amendable to genome modification, we expressed Rep78, an AAV2-derived protein with AAVS1-specific endonuclease activity, in iPS cells after adenoviral gene transfer. We showed that Rep78 efficiently associated with the AAVS1 site and triggered genome modifications within this site. On the other hand, binding to and modification of the CCR5-ZFN site by a ZFN was relatively inefficient. Our data suggest a critical influence of chromatin structure on efficacy of site-specific endonucleases used for genome editing.

The evolution of inquilinism, host-plant use and mitochondrial substitution rates in Tamalia gall aphids.

We used mitochondrial DNA data to infer phylogenies for 28 samples of gall-inducing Tamalia aphids from 12 host-plant species, and for 17 samples of Tamalia inquilinus, aphid 'inquilines' that obligately inhabit galls of the gall inducers and do not form their own galls. Our phylogenetic analyses indicate that the inquilines are monophyletic and closely related to their host aphids. Tamalia coweni aphids from different host plants were, with one exception, very closely related to one another. By contrast, the T. inquilinus aphids were strongly genetically differentiated among most of their host plants. Comparison of branch lengths between the T. coweni clade and the T. inquilinus clade indicates that the T. inquilinus lineage evolves 2.5-3 times faster for the cytochrome oxidase I gene. These results demonstrate that: (1) Tamalia inquilines originated from their gall-inducing hosts, (2) communal (multi-female) gall induction apparently facilitated the origin of inquilinism, (3) diversification of the inquilines has involved rapid speciation along host-plant lines, or the rapid evolution of host-plant races, and (4) the inquilines have undergone accelerated molecular evolution relative to their hosts, probably due to reduced effective population sizes. Our findings provide insight into the behavioural causes and evolutionary consequences of transitions from resource generation to resource exploitation.

Chemical oxygen demand analysis of wastewater using trivalent manganese oxidant with chloride removal by sodium bismuthate pretreatment.

Current chemical oxygen demand (COD) analyses generate wastes containing hexavalent and trivalent chromium, mercury, and silver. Waste disposal is difficult, expensive, and poses environmental hazards. A new COD test is proposed that eliminates these metals and shortens analysis time, where trivalent manganese oxidant replaces hexavalent chromium (dichromate). A silver catalyst is not required. Optional pretreatment removes chloride via oxidation to chlorine using sodium bismuthate, eliminating the need for mercury. Sample aqueous and solid components are separated for chloride removal, then recombined for total COD measurement. Soluble and nonsoluble COD can be determined separately. Digestion at 150 degrees C is complete in 1 hour. Results are determined by titration or by spectrophotometric reading. Test wastes contain none of the metals regulated for disposal under the Resource Conservation and Recovery Act. Results are shown for selected organic compounds and various wastewaters. Statistical comparisons are made with dichromate COD and biochemical oxygen demand (BOD5) test values.

Natural history of choroidal neovascularization in high myopia.

High myopia, or pathologic myopia, usually refers to a condition in which individuals have greater than 6 diopters of myopia or an axial length greater than 26 to 27 mm. The natural history of choroidal neovascularization in high myopia is variable, and reports to date have some conflicting information, but analysis shows fairly poor final visual outcomes. This outcome complicates recommendations for treatment and must be considered in future studies aimed at managing this disease.

Soft phonation in the male singing voice: a preliminary study.

Sustained high notes, diminishing gradually from the loudest to the softest phonation within a maneuver called messa di voce, are examined in two contrasting professional tenor voices. Signals of the sound pressure level, electroglottograph, and mean esophageal pressure are recorded, and similar maneuvers by the same subjects are examined stroboscopically. The lyric voice is found to make a gradual diminuendo while maintaining nearly constant posture of the vocal tract together with a phase of complete closure in the glottal cycle. The robust voice, by contrast, passes abruptly from a production of high subglottal pressure and a high closed quotient to one of low pressure and incomplete closure, and the transition is marked by a sudden opening of the previously constricted laryngeal collar. It is proposed that the mode of soft voice production demonstrated by the robust voice be recognized as a distinct register of the singing voice.

Psychosocial aspects of cancer genetics: women at high risk for breast and ovarian cancer.

In the past five years the advent of cancer genetic testing has created concern about the negative psychosocial sequelae of genetic counseling and testing. Research indicates that the women most likely to seek genetic testing are anxious about carrying a gene mutation and developing breast cancer. Women who are at high risk have poor knowledge and the expectation of being a gene-mutation carrier. High levels of distress have been shown to interfere with decision-making about genetic testing. Further, individuals who decline genetic testing may be at increased risk for depressive symptoms even more than those who are found to be gene-mutation carriers. There is great concern that inappropriate candidates will seek genetic testing. Improved education and access to genetic counseling are essential to help women make appropriate decisions about genetic testing. Strategies for the prevention of breast and ovarian cancer are explored, and methods to reduce the adverse psychosocial effects of decision-making about genetic testing and preventive treatment strategies are suggested.

Corticosteroids as an antiangiogenic agent for histoplasmosis-related subfoveal choroidal neovascularization.

The purpose of this study was to evaluate the role of corticosteroids in managing subfoveal choroidal neovascularization (CNV) secondary to the presumed ocular histoplasmosis syndrome. The cases of eighteen patients with histoplasmosis-related subfoveal CNV treated with corticosteroids were reviewed. Ten patients received oral prednisone for 4 to 6 weeks, and eight received a single sub-Tenon's injection of triamcinalone. Visual acuity outcomes were analyzed along with side effect profiles. At two-week follow-up, the prednisone group showed a median improvement in Snellen visual acuity of +2.0 lines, while the triamcinalone group remained essentially stable with a 0.5 line median loss. At treatment end (4 to 6 weeks), both groups showed no significant change in median acuity at 0.0 and -1.0 lines, respectively. Median final vision at 3 months also remained essentially stable at -0.5 lines for each group. Three patients reported anxiety, all of whom were taking prednisone 80 mg daily. Two patients reported increased appetite and weight gain on regimens of prednisone 80 and 100 mg daily. There were no adverse effects reported in the other patients receiving oral prednisone or in any patient receiving sub-Tenon's triamcinalone. The results suggest a beneficial effect of corticosteroids in stabilizing subfoveal CNV secondary to ocular histoplasmosis. In this small series, oral prednisone resulted in a short-term improvement in visual acuity, which stabilized over longer follow-up. The sub-Tenon's triamcinalone group achieved similar final stabilization without the initial improvement. Corticosteroids may be particularly valuable in managing neovascularization in patients who are awaiting interventions currently under development, in preventing recurrence after subfoveal surgery, or in treating non-surgical candidates. Further study is warranted to define the precise role of corticosteroids in this condition.

On pitch jumps between chest and falsetto registers in voice: data from living and excised human larynges.

The paper offers a new concept of studying abrupt chest-falsetto register transitions (jumps) based on the theory of nonlinear dynamics. The jumps were studied in an excised human larynx and in three living subjects (one female and two male). Data from the excised larynx revealed that a small and gradual change in tension of the vocal folds can cause an abrupt change of register and pitch. This gives evidence that the register jumps are manifestations of bifurcations in the vocal-fold vibratory mechanism. A hysteresis was observed; the upward register jump occurred at higher pitches and tensions than the downward jump. Due to the hysteresis, the chest and falsetto registers can be produced with practically identical laryngeal adjustments within a certain range of longitudinal tensions. The magnitude of the frequency jump was measured as the "leap ratio" F0F:F0C (fundamental frequency of the falsetto related to that of the chest register) and alternatively expressed as a corresponding musical interval, termed the "leap interval." Ranges of this leap interval were found to be different for the three living subjects (0-5 semitones for the female, 5-10 and 10-17 for the two males, respectively). These differences are considered to reflect different biomechanical properties of the vocal folds of the examined subjects. A small magnitude of the leap interval was associated with a smooth chest-falsetto transition in the female subject.

DNA repair and mutagen sensitivity in patients with triple primary cancers.

The purpose of this study was to measure DNA repair capacity and mutagen sensitivity in patients who have had three or more primary forms of cancer. It was hypothesized that, if abnormalities in DNA repair and mutagen sensitivity were cancer susceptibility factors, such findings would be seen with regularity in individuals with multiple primary cancers. DNA repair capacity was measured by determining repair of UV-irradiated plasmid DNA (pCMVCAT) transfected into peripheral blood lymphocytes. Results from 18 patients and a like number of age- and sex-matched controls demonstrated a significant difference in DNA repair capacity (P < 0.0001; odds ratio = 14). Mutagen sensitivity was measured by determining the mean number of chromatid breaks per cell after in vitro exposure to either bleomycin or 4-nitroquinoline-1-oxide. The difference in mean bleomycin- or 4-nitroquinoline-1-oxide-induced mutagen sensitivity between cases and controls was not statistically significant. Fourteen of the 18 patients had positive family histories of cancer; in 10, the history was compatible with cancer susceptibility syndromes. Although the numbers were small, there was no suggestion in this study that treatment or the presence of cancer was the cause of the DNA repair abnormalities encountered. These findings support the concept of diminished DNA repair capacity as an underlying feature in the development of a mutator phenotype.

Comparison of vocal tract formants in singing and nonperiodic phonation.

The skilled use of nonperiodic phonation techniques in combination with spectrum analysis has been proposed here as a practical method for locating formant frequencies in the singing voice. The study addresses the question of the degree of similarity between sung phonations and their nonperiodic imitations, with respect to both frequency of the first two formants as well as posture of the vocal tract. Using magnetic resonance imaging (MRI), linear predictive coding (LPC), and spectrum analysis, two types of nonperiodic phonation (ingressive and vocal fry) are compared with singing phonations to determine the degree of similarity/difference in acoustic and spatial dimensions of the vocal tract when these phonation types are used to approximate the postures of singing. In comparing phonation types, the close similarity in acoustic data in combination with the relative dissimilarity in spatial data indicates that the accurate imitations are not primarily the result of imitating the singing postures, but have instead an aural basis.

Failure to prescribe warfarin to patients with nonrheumatic atrial fibrillation.

OBJECTIVE: To determine how often warfarin was prescribed to patients with nonrheumatic atrial fibrillation in our community in 1992 when randomized trials had demonstrated that warfarin could prevent stroke with little increase in the rate of hemorrhage, and to determine whether warfarin was prescribed less frequently to older patients-the patients at highest risk of stroke but of most concern to physicians in terms of the safety of warfarin. DESIGN: Cross-sectional study. Appropriateness of warfarin was classified for each patient based on the independent judgments of three physicians applying relevant evidence and guidelines. SETTING: Two teaching hospitals and five community-based practices. PATIENTS: Consecutive patients with nonrheumatic atrial fibrillation (n = 189). MEASUREMENTS AND MAIN RESULTS: Warfarin was prescribed to 44 (23%) of the 189 patients. Warfarin was judged appropriate in 98 patients (52%), of whom 36 (37%) were prescribed warfarin. Warfarin was prescribed to 11 (14%) of 76 patients aged 75 years or older with hypertension, diabetes mellitus, or past stroke, the group at highest risk of stroke. In a multivariable logistic regression model controlling for appropriateness of warfarin and other patient characteristics, patients aged 75 years or older were less likely than younger patients to be treated with warfarin (odds ratio 0.25; 95% confidence interval 0.10, 0.65). CONCLUSIONS: Warfarin was prescribed infrequently to these patients with nonrheumatic atrial fibrillation, especially the older patients and even the patients for whom warfarin was judged appropriate. These findings indicate a substantial opportunity to prevent stroke.

Why isn't warfarin prescribed to patients with nonrheumatic atrial fibrillation?

OBJECTIVE: To determine the opinions of selected physicians in our community about use of warfarin for patients with nonrheumatic atrial fibrillation, and to determine the relation of the physicians' opinions to their practices. DESIGN: Survey of physicians, using eight hypothetical clinical vignettes to characterize physicians' opinions about use of warfarin in patients with nonrheumatic atrial fibrillation, according to patient age, risk of bleeding, and risk of stroke. SETTING: Two teaching hospitals and five community-based practices. PARTICIPANTS: Eighty physicians who cared for 189 consecutive patients with nonrheumatic atrial fibrillation. MEASUREMENTS AND MAIN RESULTS: The survey response rate was 73%. Nearly all responding physicians (90%) recommended warfarin for at least one vignette. However, physicians recommended warfarin less often for vignettes depicting 85-year-old patients than for matched vignettes depicting 65-year-old patients (odds ratio [OR] 0.03; 95% confidence interval [CI] 0.01, 0.08), and less often for cases with specified risk factors for bleeding than for matched cases without the risk factors (OR 0.01; 95% CI 0.004, 0.03); warfarin was recommended more often for cases with a recent stroke than for matched cases without this history (OR 8.2; 95% CI 3.6, 18). In practice, warfarin was prescribed more often (p < or = .05) by physicians reporting good personal experience and by those who had favorable opinions about its use. However, even physicians with good experience and favorable opinions did not prescribe warfarin to half of their patents for whom warfarin was independently judged appropriate. CONCLUSIONS: Physicians' opinions frequently opposed warfarin for older patients with nonrheumatic atrial fibrillation, and for those with bleeding risk factors. Physicians' opinions, as well as other barriers to warfarin therapy, most likely contribute to its infrequent prescription.

Standardized laryngeal videostroboscopic rating: differences between untrained and trained male and female subjects, and effects of varying sound intensity, fundamental frequency, and age.

To determine the influence of the factors gender, vocal training, sound intensity, pitch, and aging on vocal function, videolaryngostroboscopic images of 214 subjects, subdivided according to gender and status of vocal training, were evaluated by three judges with standardized rating scales, comprising aspects of laryngeal appearance (larynx/pharynx ratio; epiglottal shape; asymmetry arytenoid region; compensatory adjustments; thickness, width, length, and elasticity of vocal folds) and glottal functioning (amplitudes of excursion; duration, percentage, and type of vocal fold closure; phase differences; location of glottal chink). The video registrations were made while the subjects performed a set of phonatory tasks, comprising the utterance of the vowel /i/ at three levels of both fundamental frequency and sound intensity. Analysis of the rating scales showed generally sufficient agreement among judges. With the exception of more frequently observed complete closure and lateral phase differences of vocal fold excursions in trained subjects, no further differences were established between untrained and trained subjects. With an alpha level of p = 0.005, men differed from women with respect to laryngeal appearance (larynx/pharynx ratio, compensatory adjustments, and the presence of omega and deviant-shaped epiglottises), and their vocal folds were rated thicker in the vertical dimension, smaller in the lateral dimension, longer, and more tense, with smaller amplitudes of excursion during vibration. Glottal closure in male subjects was rated more complete, but briefer in duration. Significant effects of the factors pitch, sound intensity, and age on vocal fold appearance and glottal functioning were ascertained. Awareness of the influence of these factors, as well as the factor gender, on the rated scales is essential for an adequate evaluation of laryngostroboscopic images.

A subharmonic vibratory pattern in normal vocal folds.

This study observes in detail an F0/2 (sounding an octave below an original tone) subharmonic vibratory pattern produced in a normal larynx. Simultaneous electroglottographic and photoglottographic measurements reveal two different open phases within a subharmonic cycle- the first shorter with a simple shape, the second longer with a shape containing a "ripple." Such parameters as the large open quotient (ca. 0.8) and the high airflow values (ca. 1000 cm3/s) distinguish this phonation from the vocal fry (pulse) register. Using an electronic divider to track the subharmonic frequency, a method has been developed to observe the subharmonic vibration of the vocal folds stroboscopically. The stroboscopic visualization reveals an unusual mucosal movement during the "ripple," characterized by an opening movement of the upper margins, which interrupts the closing movement of the vocal folds. An explanation is offered that this vibratory pattern arises as a consequence of detuning of the usually identical frequencies of the dominant modes of the vocal folds, with 3:2 entrainment replacing the normal 1:1 pattern.

Differences in phonetogram features between male and female subjects with and without vocal training.

Singing not only requires good voluntary control over phonation and a musical ear, it also demands certain capacities of the voice source. These capacities include a desirable range of sound intensity and frequency, which can be measured and represented in a phonetogram. The influence of specific factors on voice capacities may be ascertained by the analysis of phonetograms. To determine the influence of the factors gender and voice training, phonetograms of 224 subjects, subdivided accordingly into four groups, were analyzed in two different ways. One is based on the rescaling of phonetograms, whereas the other derives analytic variables from the features' shape, area, and dynamic range. Analysis showed that, regarding gender, male subjects are able to produce softer phonations, whereas female subjects produced louder phonations at specific parts of their comparable frequency ranges. Trained subjects have a larger enclosed area of the phonetogram, which is primarily based on extended soft voice capabilities in both genders and the significantly larger frequency range in trained female subjects. The shape analysis, performed with Fourier Descriptors, revealed differences for the factors gender and training.

Measurement of formant frequencies and bandwidths in singing.

That singers under certain circumstances adjust the articulation of the vocal tract (formant tuning) to enhance acoustic output is both apparent from measurements and understood in theory. The precise effect of a formant on an approaching (retreating) harmonic as the latter varies in frequency during actual singing, however, is difficult to isolate. In this study variations in amplitude of radiated sound components as well as supraglottal and subglottal (esophageal) pressures accompanying the vibrato-related sweep of voice harmonics were used as a basis for estimating the effective center frequencies and bandwidths of the first and second formants.