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PubMed2003

BARCODE-ALL: accelerated and cost-effective genetic risk stratification in acute leukemia using spectrally addressable liquid bead microarrays.

An increasing number of risk-stratifying genetic lesions in acute leukemia are being discovered and characterized. To translate this important and increasing volume of information from the research laboratory into effective clinical care, however, new, fast and comprehensive assays are needed. Toward this end, we have developed a two-stage multiplexing assay of broad applicability, which combines multiplex polymerase chain reaction with multiplex detection on spectrally addressable liquid bead microarrays. Using pediatric lymphoblastic leukemia as a model system, we demonstrate that all seven of the fusion transcripts resulting from risk-stratifying chromosomal translocations can be assayed in a single well of a 96-well multiplate with 100% specificity and sensitivity, within 6 h of specimen collection. The assay is automatic and high throughput and represents a significant improvement over previously available assays targeting the same genetic changes. We conclude that user-defined assays that multiplex both target selection and detection may have broad applicability in the management of hematological malignancies.

Automation↗
PubMed2003

Electronically addressable SnO2 inverted opal gas sensors fabricated on interdigitated gold microelectrodes.

Electronically addressable thin films of tin oxide gas sensors with well-defined opaline microstructures and reproducible sensor-to-sensor responses have been fabricated on interdigitated gold microelectrodes through self-assembly growth of a monodisperse polystyrene latex film onto the electrodes followed by infiltration of tin tert-butoxide and calcination of the film.

Journal Article↗
PubMed2003

Questioning the "big assumptions". Part I: addressing personal contradictions that impede professional development.

BACKGROUND: The ultimate success of recent medical curriculum reforms is, in large part, dependent upon the faculty's ability to adopt and sustain new attitudes and behaviors. However, like many New Year's resolutions, sincere intent to change may be short lived and followed by a discouraging return to old behaviors. Failure to sustain the initial resolve to change can be misinterpreted as a lack of commitment to one's original goals and eventually lead to greater effort expended in rationalizing the status quo rather than changing it. OBJECTIVE: The present article outlines how a transformative process that has proven to be effective in managing personal change, Questioning the Big Assumptions, was successfully used in an international faculty development program for medical educators to enhance individual personal satisfaction and professional effectiveness. This process systematically encouraged participants to explore and proactively address currently operative mechanisms that could stall their attempts to change at the professional level. CONCLUSIONS: The applications of the Big Assumptions process in faculty development helped individuals to recognize and subsequently utilize unchallenged and deep rooted personal beliefs to overcome unconscious resistance to change. This approach systematically led participants away from circular griping about what was not right in their current situation to identifying the actions that they needed to take to realize their individual goals. By thoughtful testing of personal Big Assumptions, participants designed behavioral changes that could be broadly supported and, most importantly, sustained.

Attitude↗
PubMed2003

Effects of various N-terminal addressing signals on sorting and folding of mammalian CYP11A1 in yeast mitochondria.

Topogenesis of cytochrome p450scc, a resident protein of the inner membrane of adrenocortical mitochondria, is still obscure. In particular, little is known about the cause of its tissue specificity. In an attempt to clarify this point, we examined the process in Saccharomyces cerevisiae cells synthesizing cytochrome p450scc as its native precursor (pCYP11A1) or versions in which its N-terminal addressing presequence had been replaced with those of yeast mitochondrial proteins: CoxIV(1-25) and Su9(1-112). We found the pCYP11A1 and CoxIV(1-25)-mCYP11A1 versions to be effectively imported into yeast mitochondria and subjected to proteolytic processing. However, only minor portions of the imported proteins were incorporated into mitochondrial membranes, whereas their bulk accumulated as aggregates insoluble in 1% Triton X-100. Along with previously published data, this suggests that a distinguishing feature of the import of the CYP11A1 precursors into yeast mitochondria is their easy translocation into the matrix where the foreign proteins mainly undergo proteolysis or aggregation. The fraction of CYP11A1 that happens to be inserted into the inner mitochondrial membrane is effectively converted into the catalytically active holoenzyme. Experiments with the Su9(1-112)-mCYP11A1 construct bearing a re-export signal revealed that, after translocation of the fused protein into the matrix and its processing, the Su9(67-112) segment ensures association of the mCYP11A1 body with the inner membrane, but proper folding of the latter does not take place. Thus it can be said that the most specific stage of CYP11A1 topogenesis in adrenocortical mitochondria is its confinement and folding in the inner mitochondrial membrane. In yeast mitochondria, only an insignificant portion of the imported CYP11A1 follows this mechanism.

Animals↗
PubMed1999

Addressing adverse events through clinical indicators.

In an attempt to improve the reporting rate of adverse drug reactions the Adverse Drug Reaction Advisory Committee approached the Australian Council on Healthcare Standards Care Evaluation Program to develop a set of indicators to improve healthcare standards by heightening awareness amongst clinical staff of the morbidity, mortality and financial implications of adverse drug reactions. Ten clinical indicators addressing: (i) reporting of adverse drug reactions; (ii) adherence to treatment protocols for anaphylaxis; (iii) monitoring of warfarin; and (iv) monitoring of streptokinase, were field tested in ten Australian health-care organizations, to determine that the data were available, that the indicators were relevant to clinical practice and that the measures were achievable. Based on the results of this field test, six adverse drug reaction clinical indicators will be introduced into the Australian Council on Healthcare Standards Evaluation and Quality Improvement Program from January 1999.

Adverse Drug Reaction Reporting Systems↗
PubMed1998

Hermeneutic philosophy. Part II: A nursing research example of the hermeneutic imperative to address forestructures/pre-understandings.

Hermeneutic research requires that pre-understandings are brought to consciousness in order to provide the phenomenon under investigation with the greatest opportunity to reveal itself. This hermeneutic imperative is dealt with in the present study. My research involved explicating the practice knowledge of nursing on residential adolescent mental health units, and as I had worked on such units I held pre-understandings that would influence the research. I addressed pre-understandings in three ways: (i) by developing understandings of practice knowledge through a hermeneutic conceptualisation of it; (ii) by working out forestructures of the phenomenon under investigation. Using Heidegger's writing on forestructures, 20 statements were generated, interpreted and reconceptualised as my forestructures of residential adolescent mental health nursing. I used this work to review participant interviews in an effort to prevent myself from only finding what I already assumed I would find in relation to practice knowledge; and (iii) by formulating pre-understandings of the phenomenon of interest. Using Gadamer's writing on reflection, seven stories were written and interpreted as my pre-understandings of adolescent mental health nursing practice. I used this work to consider the presence of pre-understandings during textual interpretation in an attempt to prevent premature interpretive closure. In these ways, I brought my forestructures/pre-understandings to consciousness, reflected on them, questioned their origins, adequacy and legitimacy and thereby took account of their influence on myself and the research.

Adolescent↗
PubMed2001

Conserving the vitality of suffering: addressing family constraints to illness conversations.

When persons are confronted with life-threatening or chronic illness, there is always a possibility that family members other than the person experiencing the illness also suffer as they attempt to manage their own distress. This paper describes exemplars from a hermeneutic study that explored therapeutic conversations between nurses and families who were living with a member experiencing ischaemic heart disease. These conversations uncovered the complexity of both individual and family suffering following acute cardiac illness events, and called attention to nursing practices that addressed some of the ways that family members were constrained from having conversations about the illness. When spaces were created for these conversations to occur, possibilities for making sense of illness and suffering were revealed as vital. These understandings support the efforts of family members to sustain and conserve family relationships in the face of illness and suffering, and inform both the professional and personal lives of nurses.

Adaptation, Psychological↗
PubMedDate not supplied

In harm's way: recognizing and addressing alcohol risk for rural disadvantaged pregnant mothers.

The following article describes the efforts of a group of rural Vermont maternal child health (MCH) nurses who addressed alcohol risk for the pregnant mothers they served through the Women, Infants, and Children (WIC) Supplemental Nutrition program. As front-line providers, MCH nurses are in a position to both improve the detection of problem drinking during pregnancy and to strengthen maternal alcohol prevention and intervention initiatives. Results showed that a more refined screening for alcohol provided an opportunity to promote harm reduction.

Alcohol Drinking↗
PubMed2005

[Proteasomal degradation: from addressing of substrates to therapeutical perspectives].

The proteasome is the main intracellular proteolytic machinery. It is involved in all major cellular functions and decisions. It has long been thought that prior ubiquitinylation of almost all of its substrates was necessary for degradation. It has also long been considered that ubiquitinylation and degradation were two uncoupled mechanisms and that the recruitment of ubiquitinylated species was only performed by specialized subunits of the proteasome. The recent literature questions this simplified view. It also suggests that, on the one hand, the fraction of proteins hydrolyzed by the proteasome independently of their ubiquitinylation has largely been underestimated and, on the other hand, that the recognition of ubiquitinylated proteins involves complex addressing systems. Furthermore, it indicates a higher order structuration of the ubiquitin/proteasome pathway, a fraction of the proteasome and of ubiquitinylation enzymes being engaged in supramolecular complexes. Finally, proteasomal degradation is altered in a number of pathological situations. It, thus, constitutes a therapeutic target and the first applications are emerging.

Animals↗
PubMed2001

Addressing domestic violence through maternity services: policy and practice.

OBJECTIVE: to explore current policies and practices in maternity units that aim to identify, assess and support women experiencing domestic violence. DESIGN AND SETTING: a postal survey, conducted between June and October 1999, of all NHS Trusts in England and Wales that provided maternity services. PARTICIPANTS: Heads of Midwifery or the midwife with expertise or interest in domestic violence in each Trust. MAIN OUTCOME MEASURES: use of written policies and agreed practices for identifying and referring women experiencing domestic violence, such as availability of information, routine questioning of all women and offering women an appointment without their partner. RESULTS: 87% (183) of the 211 NHS Trusts providing maternity care participated in the survey. Twelve per cent of units had written policies for identifying women experiencing domestic violence, and a further 30% had some form of agreed practice. Less than half of maternity units routinely offered women an appointment without their partner, and just over half displayed material about domestic violence in places where women receive maternity care. Only three units had undertaken audit on their domestic violence practices. CONCLUSIONS: there is considerable variability around England and Wales in policies and practices related to domestic violence. It is evident that clear guidelines for identification and referral, training, audit and the integration of domestic violence policies with child protection and other policies are necessary to fully address the issues.

Education, Nursing, Continuing↗
PubMed2002

A region addresses patient safety.

The Pittsburgh Regional Healthcare Initiative (PRHI) is a coalition of 35 hospitals, 4 major insurers, more than 30 major and small-business health care purchasers, dozens of corporate and civic leaders, organized labor, and partnerships with state and federal government all working together to deliver perfect patient care throughout Southwestern Pennsylvania. PRHI believes that in pursuing perfection, many of the challenges facing today's health care delivery system (eg, waste and error in the delivery of care, rising costs, frustration and shortage among clinicians and workers, financial distress, overcapacity, and lack of access to care) will be addressed. PRHI has identified patient safety (nosocomial infections and medication errors) and 5 clinical areas (obstetrics, orthopedic surgery, cardiac surgery, depression, and diabetes) as ideal starting points. In each of these areas of work, PRHI partners have assembled multifacility/multidisciplinary groups charged with defining perfection, establishing region-wide reporting systems, and devising and implementing recommended improvement strategies and interventions. Many design and conceptual elements of the PRHI strategy are adapted from the Toyota Production System and its Pittsburgh derivative, the Alcoa Business System. PRHI is in the proof-of-concept phase of development.

Health Care Coalitions↗
PubMed2006

Collection, focusing, and metering of DNA in microchannels using addressable electrode arrays for portable low-power bioanalysis.

Although advances in microfluidic technology have enabled increasingly sophisticated biosensing and bioassay operations to be performed at the microscale, many of these applications employ such small amounts of charged biomolecules (DNA, proteins, and peptides) that they must first be preconcentrated to a detectable level. Efficient strategies for precisely handling minute quantities of biomolecules in microchannel geometries are critically needed; however, it has proven challenging to achieve simultaneous concentration, focusing, and metering capabilities with current-generation sample-injection technology. By using microfluidic chips incorporating arrays of individually addressable microfabricated electrodes, we demonstrate that DNA can be sequentially concentrated, focused into a narrow zone, metered, and injected into an analysis channel. This technique transports charged biomolecules between active electrodes upon application of a small potential difference (1 V) and is capable of achieving orders of magnitude concentration increases within a small device footprint. The collected samples are highly focused, with sample zone size and shape defined solely by electrode geometry.

DNA↗
PubMed2000

In response to the 1998 APA Division 40 Presidential Address: so where's the beef?

During the 1998 Division 40 presidential address by Linas Bieliauskas in San Francisco, several assertions were made contending that professional schools of psychology (PSP) had a deleterious effect on clinical neuropsychology by lowering professional standards, removing science from the curriculum, and producing mediocre clinicians. Bieliauskas reported to the author that he had based his opinion, in part, on data indicating that Boulder model graduates attained superior mean scores on the Examination for the Professional Practice in Psychology (EPPP) as compared to their Vail model counterparts. In order to determine whether significant differences existed between the two groups, a multivariate analysis of variance (MANOVA) was conducted using the 1997 EPPP scores reported by the Association of State and Provincial Psychology Boards (ASPPB, 1997). The author also provides a response to Bieliauskas' (1998) assertions.

Analysis of Variance↗
PubMed2005

Nutrition education for illiterate children in southern Madagascar--addressing their needs, perceptions and capabilities.

OBJECTIVE: A Knowledge, Attitudes and Practices (KAP) study was conducted in three villages of Bekily District in southern Madagascar prior to the implementation of a health education programme with children. The participatory learning concept of the Child-to-Child approach was followed to involve the children in the planning and implementation of the programme, which was covered by the German Agency for Technical Cooperation. DESIGN: To this effect, qualitative research methods such as Participatory Learning and Action techniques (focus group discussions, mapping and matrix ranking, etc.) were applied. SUBJECTS: The survey was conducted between August and December 1999. It involved a total of 55 school-aged children (6-14 years) along with 21 mothers and 34 fathers, representing different ethnic groups and educational backgrounds. RESULTS: The results show that children's KAP related to health and nutrition strongly reflect those of adults. They are not aware of a possible link between bad hygiene and the occurrence of diseases. According to them, diarrhoea or malaria is caused by consuming too large amounts of certain foods. Even if they know about certain elementary hygiene behaviours, they do not practise this in their everyday life. CONCLUSION: A major objective of the health education programme for children should be to tackle the discrepancy between hygiene-related knowledge and behaviour. Through the participatory study approach the children revealed their ability to contribute to the programme development. In using appropriate communication channels, the Child-to-Child health education programme is expected to influence the health behaviours of both adults and children. The health education programme should be combined with a literacy programme to address the children's desire to learn reading and writing.

Adolescent↗
PubMedDate not supplied

Addressing phonological questions with ultrasound.

Ultrasound can be used to address unresolved questions in phonological theory. To date, some studies have shown that results from ultrasound imaging can shed light on how differences in phonological elements are implemented. Phenomena that have been investigated include transitional schwa, vowel coalescence, and transparent vowels. A study of consonant cluster phonotactics is presented as an example of how ultrasound methodology can be used to examine phonological issues. Five English speakers presented with phonotactically illegal non-words (e.g., /zgomu/) typically repaired these sequences with vowel insertion (e.g., [zegomu]). Using ultrasound imaging, the production of these words is compared to legal sequences that are articulatorily similar, such as succumb and scum to assess the nature of the schwa found between /z/ and the following consonant. Results indicate that for some speakers, production of schwa in /zC/ sequences is not consistent with the phonological epenthesis of a schwa. Instead, speakers appear to be failing to sufficiently overlap the consonant gestures.

Biomechanical Phenomena↗
PubMed1997

Structural studies by high-field NMR spectroscopy of a binary-addressed complementary oligonucleotide system juxtaposing pyrene and perfluoro-azide units.

Recently, a new approach has been proposed to improve the site-specificity and efficiency of the modification of nucleic acid target sequences, the binary system of complementary-addressing nucleic acid sequences. The binary system comprises two oligonucleotides, one modified with a photosensitizing group and the other with a photoreactive group. The sites of chemical modification are arranged to bring the two chemical functions close enough together in space to allow efficient energy transfer from the photo-excited photosensitizer to an arylazide moiety which expels N2 to form a nitrene which subsequently covalently labels the target nucleic acid. Structural analysis performed by high-resolution 2D NMR spectroscopy (400 MHz and 600 MHz) are reported for the model binary system 1:2:3, where 1 is the target 12-mer pdGTATCAGTTTCT, 2 is a photoactivatable fluoroazide derivative dAGAAACp-L-Az and 3 is the photosensitizer derivative Pyr-pdTGATAC (here: Az is the p-azidotetrafluorobenzyl group, Pyr the pyrenyl-1-methylamino group, L a linker group). The assignment of oligonucleotide and modifying group protons was performed using 1H COSY, TOCSY and NOESY experiments. Comprehensive analysis of 1H NOESY spectra of 1:2:3 showed that terminal fragments of the complex [5'p-1T-2G-3A-4T-], [-21A-22T-23A-24C], [-8T-9T-10T-11C-12T] and [13A-14G-15A-15A-17A-18C-] gave a continuous set of intra- and inter-nucleotide interactions, typical of regular double-stranded B-DNA. In contrast, the central region of the complex composed of 5C, 6A, 7G, 19T and 20G nucleotide residues, nearest the Pyr and Az groups, was found to be distorted. Thus some signals from aromatic and/or sugar-ring protons of the above nucleotide residues were extremely broadened or almost absent. Moreover, some intra- and/or inter-nucleotide interactions, typical of the regular DNA duplex, were not detected for the [-5C-6A-7G-] and [-19T-20G-] regions of the tandem system. Instead of that, some cross-peaks of low-intensity between the H2 proton of the Pyr group and 7G(H1'), 7G(H2'/H2"), 7G(H3'), 4T(H2"), 4T(H4') and 4T(H5'/H5") were observed. Additional 1H -1H NOE-interactions between methylene protons of the linker group L and some sugar ring protons of 18C nucleotide residue were detected. A preliminary structural model, constructed using proton-proton distances between Pyr and the DNA and Az-L and DNA obtained from a 1H NOESY experiment at 300 ms mixing time as constraints for the refinement of the structure, displayed significant distortion from B-DNA of the double-stranded helix in the middle of the complex, (-5C-6A-7G, -18C-19T-20G-). The Pyr group was located in what remains of the minor groove near 4T, 5C, 6A and 7G and the centroid of the azide ring less than 9A degrees from the centroid of the ring system of Pyr group.

Azides↗
PubMed1999

Refined high-field NMR solution structure of a binary-addressed pyrene/perfluoro-azide complementary DNA oligonucleotide system shows extensive distortion in the central nick region.

The structural analysis of the photoactivated binary system of complementary-addressing nucleic acid sequences (1:2:3) by high-resolution NMR spectroscopy and restrained molecular dynamics is reported. The binary system comprised a 12 base-pair target DNA sequence, pdGTATCAGTTTCT (1), and two hexanucleotides, (dAGAAACp-L-Az (2) and Pyr-pdTGATAC (3)), complementary to neighbouring sites in the target DNA. Oligonucleotide (2) is conjugated with a p-azidotetrafludrobenzyl group (Az) via a linker group (L), and the other oligonucleotide (3) is equipped with the photosensitizing pyrenyl-1-methylamino group (Pyr). We now extend the structural analysis of 1:2:3, which was previously based on qualitative 2D 1H-NMR data and thermodynamic analysis of complex formation from UV-visible thermal denaturation experiments. In the current work structural refinement was performed by separate molecular dynamics runs for six different starting structures based on 318 proton-proton distance-range constraints, evaluated from the 1H-NOESY spectrum (tau(mix) = 200 ms, 600 MHz) using complete relaxation matrix analysis (NMR/TRIAD/MARDIGRAS). Additional Watson-Crick hydrogen bond restraints were included in the calculations based on the detected signals from the exchangeable protons, using REFOPT(NY) methods. The final averaged structure obtained from the six refined co-ordinate sets showed a considerable degree of axis bend (62.5 degrees) with the bending point in the middle of the duplex in the region of the backbone nick between the two short oligonucleotides. The complex behaves dynamically as the equivalent of two short B-DNA-like duplexes displaying a hinge-like flexing at their junction. In all final structures the Pyr function location was very restricted, the aromatic group lying in the duplex minor groove near residues 4T, 5C and 2T. In contrast, the location of the perfluoroazido group was different in all the final structures, indicating the high flexibility of this group in the duplex. The only feature common to all six final azido group orientations was the outside location on the side of the major groove. The distance between the Pyr and Az groups varied from 11 A to 24 A for the six final structures (17 A, final average structure). The dynamics of duplex denaturation for 1:2:3 was probed by monitoring the temperature-induced NMR line broadening of the imino protons in a 1D variable temperature NMR experiment. The melting of 1:2:3 starts both from the ends and from the middle part of the duplex at the backbone break between the two short oligonucleotides reflecting the destabilisation of the pyrene-arylazido nick region in the duplex.

Azides↗
PubMedDate not supplied

Ten steps to developing a national agenda to address financial conflicts of interest in industry sponsored clinical research.

Financial liaisons between clinical researchers, research institutions, and industrial sponsors have gained momentum in recent years. In the process, it has been argued by many that trust in the research infrastructure is being eroded by the financial conflicts of interest that emerge from these arrangements. Yet, the financial resources of industry are needed to continue technology transfer from the bench to the bedside. Policy makers and government regulators are currently struggling to determine how to best manage financial conflicts of interest that emerge from these liaisons. Various organizations and government entities have proposed different strategies. This paper explores the limitations of existing measures and recommends that a unified national agenda is needed. We propose 10 steps to develop an agenda to address financial conflicts of interest in industry-sponsored clinical research.

Biomedical Research↗