Development and validation of nursing students' moral sensitivity questionnaire in Taiwan.

Ethical literacy is a critical aspect of professional nursing development. It is considered an essential quality that nursing professionals should possess throughout their careers. Moral sensitivity serves as the foundation for developing ethical literacy. The objective of this study was to develop a reliable tool for assessing moral sensitivity among nursing students. The questionnaire was developed following a rigorous approach, consisting of three stages process, combining the Schwartz-Barcott and Kim hybrid model of concept development with the methodology suggested by Devellis and Waltz. A total of 297 nursing students (287 females, 10 males; mean age: 18.7 years) participated in the study, with five invalid questionnaires excluded from the analysis. The questionnaire's reliability was established through internal consistency and test-retest reliability analyses. Furthermore, the moral sensitivity questionnaire for nursing students demonstrated satisfactory validity through the results of construct, convergent and discriminant validation procedures. The study findings revealed a significant correlation between the internship performance of students and their overall moral sensitivity score. The questionnaire would be appropriated to be included as a supplemental measure for ethical literacy evaluation.

Effectiveness of Motivational Interviewing in Regard to Activities of Daily Living and Motivation for Rehabilitation among Stroke Patients.

BACKGROUND: Stroke patients urgently need rehabilitation to enhance activities of daily living. This study aims to determine whether motivational interviewing (MI) improves the performance of activities of daily living and enhances motivation for rehabilitation among first-stroke patients. METHODS: A quasi-experimental design was used in this study. The study recruited 65 patients between March and October 2016. Before the intervention, all patients received routine care. The experimental group (n = 33) received weekly sessions of MI for 6 weeks, whereas the control group (n = 32) received individual attention from a research nurse weekly for 6 weeks. Structured questionnaires were used to collect data, including demographic data, activities of daily living data (Barthel index {BI} and instrumental activities of daily living {IADLs} scale), and rehabilitation motivation data. RESULTS: The BI and IADLs scores significantly improved with time in both the experimental and control groups. The generalized estimating equation approach showed that at 6 weeks and 3 months after the intervention, the rehabilitation motivation scores in the experimental group were respectively 3.10 and 2.54 points higher than those in the control group, with significant differences. CONCLUSIONS: MI could effectively enhance motivation for rehabilitation among stroke patients.

Nurses' perspectives on moral distress: A Q methodology approach.

BACKGROUND: Moral distress occurs when nurses experience ethical dilemmas. Issues related to these dilemmas are addressed in some nursing education courses. Nurses' reaction to dilemma such as moral distress is relatively less noticed. OBJECTIVE: This study aimed to identify and describe the various types of perceptions of moral distress exhibited by nurses. RESEARCH DESIGN: This study applied Q methodology to explore the perspectives of nurses regarding moral distress. Data were collected in two stages. First, in-depth interviews were conducted to collect nurses' opinions. Sentences that best fit the concepts of moral distress were extracted for the construction of Q statements. Second, nurses subjectively ranked these Q statements so that the relevant severity of moral distress could be determined using Q sorts. The study participants were nurses at a regional teaching hospital in northeast Taiwan. A total of 60 participants were invited to rank 40 moral distress Q statements. Ethical considerations: The study protocol was approved by the institutional review board of National Yang-Ming University Hospital. Only the participants who signed an informed consent form participated in the study. The respondents' right to withdraw from the study was respected. FINDINGS: Five types of responses were identified regarding the nurses' perspectives. These types were "conflict with personal values," "excessive of workload," "curbing of autonomy," "constraint engendered by organizational norms," and "self-expectation frustration." CONCLUSION: The findings regarding nurses' experiences of moral distress can be used to construct multifaceted policies and solutions and to incorporate ethical education in training programs.

Evaluating the effect of three teaching strategies on student nurses' moral sensitivity.

BACKGROUND: The Taiwan Nursing Accreditation Council has proposed eight core professional nursing qualities including ethical literacy. Consequently, nursing ethics education is a required course for student nurses. These courses are intended to improve the ethical literacy. Moral sensitivity is the cornerstone of ethical literacy, and learning moral sensitivity is the initial step towards developing ethical literacy. OBJECTIVES: To explore the effect of nursing ethics educational interventions based on multiple teaching strategies on student nurses moral sensitivity. Based on the visual, auditory and kinaesthetic model, three strategies were developed for determining the programme components and corresponding learning styles. RESEARCH DESIGN: This was a quasi-experimental study. PARTICIPANTS: A total of 234 junior-college student nurses participated in this study. All participants were aged 18-19 years. Ethical considerations: The study protocol was approved by the institutional review boards of Kaohsiung Veterans General Hospital. Only the participants who signed an informed consent form took part in the study. The participants were permitted to withdraw from the study at any point if they wished to do so without affecting their academic score. RESULTS: The scores of Modified Moral Sensitivity Questionnaire for Student Nurses were significantly improved after the intervention of integrating multiple teaching strategies ( p = .042). Significant relationships were observed between the satisfaction scores of two teaching strategies and moral sensitivity. The results indicated that using multiple teaching strategies is effective for promoting nursing ethics learning. CONCLUSION: This strategy was consistent with the student nurses' preferred learning style and was used to correct their erroneous ethical conceptions, assisting in developing their ethical knowledge.

Fluorescent saxitoxins for live cell imaging of single voltage-gated sodium ion channels beyond the optical diffraction limit.

A desire to better understand the role of voltage-gated sodium channels (Na(V)s) in signal conduction and their dysregulation in specific disease states motivates the development of high precision tools for their study. Nature has evolved a collection of small molecule agents, including the shellfish poison (+)-saxitoxin, that bind to the extracellular pore of select Na(V) isoforms. As described in this report, de novo chemical synthesis has enabled the preparation of fluorescently labeled derivatives of (+)-saxitoxin, STX-Cy5, and STX-DCDHF, which display reversible binding to Na(V)s in live cells. Electrophysiology and confocal fluorescence microscopy studies confirm that these STX-based dyes function as potent and selective Na(V) labels. The utility of these probes is underscored in single-molecule and super-resolution imaging experiments, which reveal Na(V) distributions well beyond the optical diffraction limit in subcellular features such as neuritic spines and filopodia.

The double-helix microscope super-resolves extended biological structures by localizing single blinking molecules in three dimensions with nanoscale precision.

The double-helix point spread function microscope encodes the axial (z) position information of single emitters in wide-field (x,y) images, thus enabling localization in three dimensions (3D) inside extended volumes. We experimentally determine the statistical localization precision σ of this approach using single emitters in a cell under typical background conditions, demonstrating σ < 20 nm laterally and <30 nm axially for N ≈ 1180 photons per localization. Combined with light-induced blinking of single-molecule labels, we present proof-of-concept imaging beyond the optical diffraction limit of microtubule network structures in fixed mammalian cells over a large axial range in three dimensions.

Modeling Parkinson's disease using induced pluripotent stem cells.

Our understanding of the underlying molecular mechanism of Parkinson's disease (PD) is hampered by a lack of access to affected human dopaminergic (DA) neurons on which to base experimental research. Fortunately, the recent development of a PD disease model using induced pluripotent stem cells (iPSCs) provides access to cell types that were previously unobtainable in sufficient quantity or quality, and presents exciting promises for the elucidation of PD etiology and the development of potential therapeutics. To more effectively model PD, we generated two patient-derived iPSC lines: a line carrying a homozygous p.G2019S mutation in the leucine-rich repeat kinase 2 (LRRK2) gene and another carrying a full gene triplication of the α-synuclein encoding gene, SNCA. We demonstrated that these PD-linked pluripotent lines were able to differentiate into DA neurons and that these neurons exhibited increased expression of key oxidative stress response genes and α-synuclein protein. Moreover, when compared to wild-type DA neurons, LRRK2-G2019S iPSC-derived DA neurons were more sensitive to caspase-3 activation caused by exposure to hydrogen peroxide, MG-132, and 6-hydroxydopamine. In addition, SNCA-triplication iPSC-derived DA neurons formed early ubiquitin-positive puncta and were more sensitive to peak toxicity from hydrogen peroxide-induced stress. These aforementioned findings suggest that LRRK2-G2019S and SNCA-triplication iPSC-derived DA neurons exhibit early phenotypes linked to PD. Given the high penetrance of the homozygous LRRK2 mutation, the expression of wild-type α-synuclein protein in the SNCA-triplication line, and the clinical resemblance of patients afflicted with these familial disorders to sporadic PD patients, these iPSC-derived neurons may be unique and valuable models for disease diagnostics and development of novel pharmacological agents for alleviation of relevant disease phenotypes.

Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores.

Superresolution imaging techniques based on sequential imaging of sparse subsets of single molecules require fluorophores whose emission can be photoactivated or photoswitched. Because typical organic fluorophores can emit significantly more photons than average fluorescent proteins, organic fluorophores have a potential advantage in super-resolution imaging schemes, but targeting to specific cellular proteins must be provided. We report the design and application of HaloTag-based target-specific azido DCDHFs, a class of photoactivatable push-pull fluorogens which produce bright fluorescent labels suitable for single-molecule superresolution imaging in live bacterial and fixed mammalian cells.

Single-molecule spectroscopy and imaging of biomolecules in living cells.

The number of reports per year on single-molecule imaging experiments has grown roughly exponentially since the first successful efforts to optically detect a single molecule were completed over two decades ago. Single-molecule spectroscopy has developed into a field that includes a wealth of experiments at room temperature and inside living cells. The fast growth of single-molecule biophysics has resulted from its benefits in probing heterogeneous populations, one molecule at a time, as well as from advances in microscopes and detectors. This Perspective summarizes the field of live-cell imaging of single biomolecules.

Azido push-pull fluorogens photoactivate to produce bright fluorescent labels.

Dark azido push-pull chromophores have the ability to be photoactivated to produce bright fluorescent labels suitable for single-molecule imaging. Upon illumination, the aryl azide functionality in the fluorogens participates in a photochemical conversion to an aryl amine, thus restoring charge-transfer absorption and fluorescence. Previously, we reported that one compound, DCDHF-V-P-azide, was photoactivatable. Here, we demonstrate that the azide-to-amine photoactivation process is generally applicable to a variety of push-pull chromophores, and we characterize the photophysical parameters including photoconversion quantum yield, photostability, and turn-on ratio. Azido push-pull fluorogens provide a new class of photoactivatable single-molecule probes for fluorescent labeling and super-resolution microscopy. Lastly, we demonstrate that photoactivated push-pull dyes can insert into bonds of nearby biomolecules, simultaneously forming a covalent bond and becoming fluorescent (fluorogenic photoaffinity labeling).

DCDHF fluorophores for single-molecule imaging in cells.

There is a persistent need for small-molecule fluorescent labels optimized for single-molecule imaging in the cellular environment. Application of these labels comes with a set of strict requirements: strong absorption, efficient and stable emission, water solubility and membrane permeability, low background emission, and red-shifted absorption to avoid cell autofluorescence. We have designed and characterized several fluorophores, termed "DCDHF" fluorophores, for use in live-cell imaging based on the push-pull design: an amine donor group and a 2-dicyanomethylene-3-cyano-2,5-dihydrofuran (DCDHF) acceptor group, separated by a pi-rich conjugated network. In general, the DCDHF fluorophores are comparatively photostable, sensitive to local environment, and their chemistries and photophysics are tunable to optimize absorption wavelength, membrane affinity, and solubility. Especially valuable are fluorophores with sophisticated photophysics for applications requiring additional facets of control, such as photoactivation. For example, we have reengineered a red-emitting DCDHF fluorophore so that it is dark until photoactivated with a short burst of low-intensity violet light. This molecule and its relatives provide a new class of bright photoactivatable small-molecule fluorophores, which are needed for super-resolution imaging schemes that require active control (here turning-on) of single-molecule emission.

A photoactivatable push-pull fluorophore for single-molecule imaging in live cells.

We have reengineered a red-emitting dicyanomethylenedihydrofuran push-pull fluorophore so that it is dark until photoactivated with a short burst of low-intensity violet light. Photoactivation of the dark fluorogen leads to conversion of an azide to an amine, which shifts the absorption to long wavelengths. After photoactivation, the fluorophore is bright and photostable enough to be imaged on the single-molecule level in living cells. This proof-of-principle demonstration provides a new class of bright photoactivatable fluorophores, as are needed for super-resolution imaging schemes that require active control of single molecule emission.

Photophysical properties of acene DCDHF fluorophores: long-wavelength single-molecule emitters designed for cellular imaging.

We report the solvatochromic, viscosity-sensitive, and single-molecule photophysics of the fluorophores DCDHF-N-6 and DCDHF-A-6. These molecules are members of the dicyanomethylenedihydrofuran (DCDHF) class of single-molecule emitters that contain an amine electron donor and a DCDHF acceptor linked by a conjugated unit; DCDHF-N-6 and DCDHF-A-6 have naphthalene- and anthracene-conjugated linkers, respectively. These molecules maintain the beneficial photophysics of the phenylene-linked DCDHF (i.e., photostability, emission wavelength dependence on solvent polarity, and quantum yield sensitivity to solvent viscosity), yet offer absorption and emission at longer wavelengths that are more appropriate for cellular imaging. We demonstrate that these new fluorophores are less photolabile in an aqueous environment than several other commonly used dyes (rhodamine 6G, Texas Red, and fluorescein). Finally, we image single copies of the acene DCDHFs diffusing in the plasma membrane of living cells.