Lessons learned from microsatellite development for nonmodel organisms using 454 pyrosequencing.

Microsatellites, also known as simple sequence repeats (SSRs), are among the most commonly used marker types in evolutionary and ecological studies. Next Generation Sequencing techniques such as 454 pyrosequencing allow the rapid development of microsatellite markers in nonmodel organisms. 454 pyrosequencing is a straightforward approach to develop a high number of microsatellite markers. Therefore, developing microsatellites using 454 pyrosequencing has become the method of choice for marker development. Here, we describe a user friendly way of microsatellite development from 454 pyrosequencing data and analyse data sets of 17 nonmodel species (plants, fungi, invertebrates, birds and a mammal) for microsatellite repeats and flanking regions suitable for primer development. We then compare the numbers of successfully lab-tested microsatellite markers for the various species and furthermore describe diverse challenges that might arise in different study species, for example, large genome size or nonpure extraction of genomic DNA. Successful primer identification was feasible for all species. We found that in species for which large repeat numbers are uncommon, such as fungi, polymorphic markers can nevertheless be developed from 454 pyrosequencing reads containing small repeat numbers (five to six repeats). Furthermore, the development of microsatellite markers for species with large genomes was also with Next Generation Sequencing techniques more cost and time-consuming than for species with smaller genomes. In this study, we showed that depending on the species, a different amount of 454 pyrosequencing data might be required for successful identification of a sufficient number of microsatellite markers for ecological genetic studies.

Fewer invited talks by women in evolutionary biology symposia.

Lower visibility of female scientists, compared to male scientists, is a potential reason for the under-representation of women among senior academic ranks. Visibility in the scientific community stems partly from presenting research as an invited speaker at organized meetings. We analysed the sex ratio of presenters at the European Society for Evolutionary Biology (ESEB) Congress 2011, where all abstract submissions were accepted for presentation. Women were under-represented among invited speakers at symposia (15% women) compared to all presenters (46%), regular oral presenters (41%) and plenary speakers (25%). At the ESEB congresses in 2001-2011, 9-23% of invited speakers were women. This under-representation of women is partly attributable to a larger proportion of women, than men, declining invitations: in 2011, 50% of women declined an invitation to speak compared to 26% of men. We expect invited speakers to be scientists from top ranked institutions or authors of recent papers in high-impact journals. Considering all invited speakers (including declined invitations), 23% were women. This was lower than the baseline sex ratios of early-mid career stage scientists, but was similar to senior scientists and authors that have published in high-impact journals. High-quality science by women therefore has low exposure at international meetings, which will constrain Evolutionary Biology from reaching its full potential. We wish to highlight the wider implications of turning down invitations to speak, and encourage conference organizers to implement steps to increase acceptance rates of invited talks.

Independence among physiological traits suggests flexibility in the face of ecological demands on phenotypes.

Phenotypic flexibility allows animals to adjust their physiology to diverse environmental conditions encountered over the year. Examining how these varying traits covary gives insights into potential constraints or freedoms that may shape evolutionary trajectories. In this study, we examined relationships among haematocrit, baseline corticosterone concentration, constitutive immune function and basal metabolic rate in red knot Calidris canutus islandica individuals subjected to experimentally manipulated temperature treatments over an entire annual cycle. If covariation among traits is constrained, we predict consistent covariation within and among individuals. We further predict consistent correlations between physiological and metabolic traits if constraints underlie species-level patterns found along the slow-fast pace-of-life continuum. We found no consistent correlations among haematocrit, baseline corticosterone concentration, immune function and basal metabolic rate either within or among individuals. This provides no evidence for constraints limiting relationships among these measures of the cardiovascular, endocrine, immune and metabolic systems in individual red knots. Rather, our data suggest that knots are free to adjust individual parts of their physiology independently. This makes good sense if one places the animal within its ecological context where different aspects of the environment might put different pressures on different aspects of physiology.

Contemporary gene flow and mating system of Arabis alpina in a Central European alpine landscape.

BACKGROUND AND AIMS: Gene flow is important in counteracting the divergence of populations but also in spreading genes among populations. However, contemporary gene flow is not well understood across alpine landscapes. The aim of this study was to estimate contemporary gene flow through pollen and to examine the realized mating system in the alpine perennial plant, Arabis alpina (Brassicaceae). METHODS: An entire sub-alpine to alpine landscape of 2 km(2) was exhaustively sampled in the Swiss Alps. Eighteen nuclear microsatellite loci were used to genotype 595 individuals and 499 offspring from 49 maternal plants. Contemporary gene flow by pollen was estimated from paternity analysis, matching the genotypes of maternal plants and offspring to the pool of likely father plants. Realized mating patterns and genetic structure were also estimated. KEY RESULTS: Paternity analysis revealed several long-distance gene flow events (≤1 km). However, most outcrossing pollen was dispersed close to the mother plants, and 84 % of all offspring were selfed. Individuals that were spatially close were more related than by chance and were also more likely to be connected by pollen dispersal. CONCLUSIONS: In the alpine landscape studied, genetic structure occurred on small spatial scales as expected for alpine plants. However, gene flow also covered large distances. This makes it plausible for alpine plants to spread beneficial alleles at least via pollen across landscapes at a short time scale. Thus, gene flow potentially facilitates rapid adaptation in A. alpina likely to be required under ongoing climate change.

Effectiveness of individualized developmental care for low-risk preterm infants: behavioral and electrophysiologic evidence.

OBJECTIVE: We assessed the effectiveness of individualized developmental support in the special care nursery for low-risk preterm infants. SETTING: A university-affiliated teaching hospital. PARTICIPANTS: Twelve healthy full-term infants, and 24 low-risk preterm infants randomly assigned to a control or an experimental group. DESIGN: The preterm control group received standard care and the preterm experimental group received individualized developmental care at the same special care nursery. OUTCOME MEASURES: Medical, behavioral (Assessment of Preterm Infants' Behavior and Prechtl's Neurological Examination of the Full-Term Newborn Infant), and electrophysiologic outcome (using quantitative electroencephalography with topographic mapping) of all three groups was assessed 2 weeks after the expected due date. RESULTS: No between- or among-group medical differences were seen for this low-risk, healthy sample. The preterm experimental group showed behavioral and electrophysiologic performances comparable to those of the full-term group, whereas the preterm control group performed significantly less well. Behavioral measures suggested significantly poorer attentional functioning for the preterm control group. Electrophysiologic results implicated the frontal lobe. CONCLUSIONS: Individualized developmental intervention supports neurobehavioral functioning as measured at 2 weeks post-term. It appears to prevent frontal lobe and attentional difficulties in the newborn period, the possible causes of behavioral and scholastic disabilities often seen in low-risk preterm infants at later ages.

Prosthetic replacement of the aortic arch.

Four patients are reported in whom the aortic arch and variable portions of the ascending and descending aorta were replaced with a prosthesis. In three patients the preoperative diagnosis was dissecting aneurysm of the aortic arch and in one an arteriosclerotic aneurysm of the aortic arch was present. A combination of surface cooling and cardiopulmonary bypass was utilized to produce total body hypothermia. Arch replacement was carried out during a period of total circulatory arrest. Cardiopulmonary bypass was then utilized to warm the patient and resuscitate the heart. The average duration of cerebral ischemia was 43 minutes and the average duration of myocardial ischemia was 74 minutes. The average lowest esophageal temperature was 14 degrees C., and the average lowest rectal temperature was 18 degrees C. Three patients are alive and well 4 to 13 months following surgery. One patient died 4 days postoperatively of pulmonary insufficiency. This experience indicates that by utilizing total body hypothermia and circulatory arrest aortic arch replacement can be carried out with an acceptable mortality rate. Corrective surgery could be offered to patients with life-threatening enlarging aneurysms of the aortic arch.

Surgical treatment of aneurysms of the inferior left ventricular wall.

Experience with surgical treatment of 10 patients with aneurysms of the inferior wall of the left ventricle is presented. Six of the 10 aneurysms were false (pseudoaneurysms), and four were classified as true aneurysms. All except one resulted from myocardial infarction. Combined procedures, performed at the time of aneurysm resection, included mitral valve replacement (five patients), coronary artery bypass grafting (four patients), and closure of an interventricular septal defect (one patient). Three of four patients with true inferior aneurysms had mitral valve dysfunction, whereas only two of six patients with false aneurysms required mitral valve replacement (one because of infective endocarditis). Nine of the 10 patients survived operation, and all are functionally improved except one. On the basis of this and previously reported experience, it is concluded that a substantial proportion of inferior left ventricular aneurysms exhibit the pathological features of false aneurysms. Because of the associated propensity toward rupture of such lesions, an aggressive surgical approach is recommended.

Informed consent and the elderly. An ethical challenge for critical care nursing.

Nursing ethics was long reticent about ethical challenges that elderly patients pose for critical care nursing. One of these, the legal doctrine of informed consent, has important implications for critical care nursing ethics. Deriving from the principle of respect for persons and intended to preserve their autonomy, informed consent represents both a duty and an ideal for care givers to implement in the process of ethical decision making. All too often the ideal is lost, however, and the doctrine reduced to a sterile and bureaucratic procedure. When elderly patients are unable to give an adequately informed consent, advance directives can enable them to express their wishes by prior choosing. If they become cognitively impaired, however, it is much more difficult to determine what, if any, preferences such a patient might express if able to do so. Medical empowerment of the elderly, a laudable social goal, can be as contradictory as informed consent itself and many elderly patients may opt out of their own decision making. The resultant moral distress of such a complex process is still another ethical challenge that faces the critical care nurse. Because nursing holds a position of moral centrality among the health care professions, critical care nurses cannot avoid the prospect that issues like those identified in this article will continue to challenge and confront them in the coming decade. By turning to colleagues in nursing as well as other professions, nurses can best strengthen and consolidate their vital role as mediators of meaning and morality in life-and-death situations.

Computerized chaplaincy--today and tomorrow.

Health care chaplaincy is not an obvious setting for "high tech" methods and equipment. Yet computer technology is rapidly revolutionizing both medicine and ministry. DRGs (diagnosis-related groups) and other innovations in health care make it imperative that chaplains be both familiar and comfortable with computer technology. Obstacles to this goal include computer "block," clergy resistance, cost factors and the myth that pastoral care requires no technology. Chaplains are urged to explore new uses for computers in ministry, while being cautious about too rapidly embracing technology as an end in itself. Each chaplain must determine for herself/himself how best to utilize computers in chaplaincy.

Protection of patient rights and welfare in the VA cooperative studies program.

Four VA Cooperative Studies Program Coordinating Centers have been established by the Cooperative Studies Program of the Medical Research Service of the Veterans Administration. Affiliated with each Coordinating Center is a Human Rights Committee (HRC). These HRCs function similarly to the Institutional Review Board but differ from it in many significant ways. This paper presents the organizational structures, the responsibilities, and the inherent dynamics of the HRCs. With clear and defined objectives and guidelines, the HRCs monitor the multicenter cooperative studies conducted within the VA health care system. The HRCs strive to balance the need for protecting patients' rights and welfare with the need for research. Its specific role is to ensure that multicenter trials are planned, implemented, and evaluated such that the rights and welfare of study subjects are protected. This is accomplished by reviewing the content and the procedures of the informed consent during the planning stage as well as by the site visits and continuing reviews during the ongoing phase of each cooperative study.