Identification of the Novel Allele HLA-DPB1*1782:01 in a Deceased Donor by Third-Generation Sequencing.
HLA-DPB1*1782:01 differs from HLA-DPB1*04:02:01:01 by a non-synonymous nucleotide substitution at codon 87.
Humans↗
Biomedical subjects
S Johnson
Publications and source records attributed to S Johnson.
Construct validation of a novel hybrid surgical simulator.
BACKGROUND: Simulated minimal access surgery has improved recently as both a learning and assessment tool. The construct validation of a novel simulator, ProMis, is described for use by residents in training. METHODS: ProMis is a surgical simulator that can design tasks in both virtual and actual reality. A pilot group of surgical residents ranging from novice to expert completed three standardized tasks: orientation, dissection, and basic suturing. The tasks were tested for construct validity. Two experienced surgeons examined the recorded tasks in a blinded fashion using an objective structured assessment of technical skills format (OSATS: task-specific checklist and global rating score) as well as metrics delivered by the simulator. RESULTS: The findings showed excellent interrater reliability (Cronbach's alpha of 0.88 for the checklist and 0.93 for the global rating). The median scores in the experience groups were statistically different in both the global rating and the task-specific checklists (p < 0.05). The scores for the orientation task alone did not reach significance (p = 0.1), suggesting that modification is required before ProMis could be used in isolation as an assessment tool. CONCLUSIONS: The three simulated tasks in combination are construct valid for differentiating experience levels among surgeons in training. This hybrid simulator has potential added benefits of marrying the virtual with actual, and of combining simple box traits and advanced virtual reality simulation.
Computer Simulation↗
Physical and cognitive task analysis in interventional radiology.
AIM: To identify, describe and detail the cognitive thought processes, decision-making, and physical actions involved in the preparation and successful performance of core interventional radiology procedures. MATERIALS AND METHODS: Five commonly performed core interventional radiology procedures were selected for cognitive task analysis. Several examples of each procedure being performed by consultant interventional radiologists were videoed. The videos of those procedures, and the steps required for successful outcome, were analysed by a psychologist and an interventional radiologist. Once a skeleton algorithm of the procedures was defined, further refinement was achieved using individual interview techniques with consultant interventional radiologists. Additionally a critique of each iteration of the established algorithm was sought from non-participating independent consultant interventional radiologists. RESULTS: Detailed task descriptions and decision protocols were developed for five interventional radiology procedures (arterial puncture, nephrostomy, venous access, biopsy-using both ultrasound and computed tomography, and percutaneous transhepatic cholangiogram). Identical tasks performed within these procedures were identified and standardized within the protocols. CONCLUSIONS: Complex procedures were broken down and their constituent processes identified. This might be suitable for use as a training protocol to provide a universally acceptable safe practice at the most fundamental level. It is envisaged that data collected in this way can be used as an educational resource for trainees and could provide the basis for a training curriculum in interventional radiology. It will direct trainees towards safe practice of the highest standard. It will also provide performance objectives of a simulator model.
Cholangiography↗
Quantitative analysis of magnetic excitations in Landau flux-closure structures using synchrotron-radiation microscopy.
We have measured the excitation spectrum of six micron magnetic squares using x-ray magnetic circular dichroism. We observe all three excitations expected in a Landau flux-closure pattern. High temporal and spatial resolution allows quantitative analysis of the excitations. A short magnetic in plane pulse excites the magnetic element and we observe precessional motion of the magnetization within the domains as well as a domain wall mode and vortex motion. The vortex moves perpendicular to the excitation field and relaxes without showing a circulating orbit.
Journal Article↗
Nonplanar photolithography with computer-generated holograms.
We outline a method for accomplishing photolithography on grossly nonplanar substrates. First we compute an approximation of the diffraction pattern that will produce the desired light-intensity distribution on the substrate to be patterned. This pattern is then digitized and converted into a format suitable for manufacture by a direct-write method. The resultant computer-generated hologram mask is then used in a custom alignment tool to expose the photoresist-coated substrate. The technique has many potential applications in the packaging of microelectronics and microelectromechanical systems.
Journal Article↗
Randomised trial of parental support for families with very preterm children: outcome at 5 years.
AIMS: To test the effectiveness of a home based developmental education intervention in improving outcome at 5 years for very preterm infants. METHODS: The Avon Premature Infant Project (APIP) is a randomised controlled trial in which the parents of 284 babies born <33 weeks gestational age received a developmental education programme, a social support intervention, or standard care. A term reference population was also recruited. This study reports outcomes at 5 years (mean age 58 months 15 days) for 187 (66%) of these children without disability. Outcomes were assessed using the British Ability Scales II for cognitive development, the Movement ABC for motor impairment, and the Child Behavior Checklist for behavioural problems. RESULTS: Preterm infants showed poorer cognitive performance than their term peers. Mean (SD) general conceptual ability (GCA) scores were: Portage 99.2 (15.7); parent adviser 100.3 (14.8); preterm control 101.1 (15.0); term reference 107.2 (13.4). There were no significant differences between preterm groups in GCA scores indicating no effect of either intervention. Similarly, there was no significant effect of intervention on behavioural or motor outcomes. Further analyses, in which outcome data were adjusted for social factors, did not reveal any differences between the three preterm groups or by subgroups classified by a range of perinatal variables. CONCLUSION: The small advantage shown at 2 years of age is no longer detectable at 5 years. These results question the effectiveness of early intervention in enhancing cognitive, behavioural, and motor function at 5 years.
Birth Weight↗
Demonstration of the appearance of the paraspinal musculoligamentous structures of the cervical spine using ultrasound.
The application of ultrasound in the imaging of the neck has primarily focussed on anterior structures (e.g., thyroid gland). Structures located on the posterior aspect of the neck have received little attention. This study illustrates the capability of modern ultrasound equipment in visualising the musculoligamentous structures of the neck, particularly the paraspinal musculature. Ten healthy adult volunteers (6 female; 4 male) underwent ultrasound examination of the cervical spine. A standardised technique for transducer placement was adopted and successive images of the neck of each subject were obtained. Spatial compound (extended field of view) images were obtained in subjects using one of two different ultrasound systems. Images of structures produced by ultrasound were compared to those achieved with magnetic resonance imaging in three subjects. Identification of key landmarks aided orientation and identification of structures. The internal architecture of the musculoligamentous structures of the cervical spine, especially the posterior neck muscles, was demonstrated well using ultrasound. Our study showed that modern ultrasound equipment is capable of producing clear images of the posterior cervical spine musculature and certain bony features.
Adult↗
Blue cone monochromatism: a phenotype and genotype assessment with evidence of progressive loss of cone function in older individuals.
AIM: To perform a detailed clinical and psychophysical assessment of the members of three British families affected with blue cone monochromatism (BCM), and to determine the molecular basis of disease in these families. METHODS: Affected and unaffected members of three families with BCM were examined clinically and underwent electrophysiological and detailed psychophysical testing. Blood samples were taken for DNA extraction. The strategy for molecular analysis was to amplify the coding regions of the long wavelength-sensitive (L) and middle wavelength-sensitive (M) cone opsin genes and the upstream locus control region by polymerase chain reaction, and to examine these fragments for mutations by direct sequencing. RESULTS: We have confirmed the reported finding of protan-like D-15 arrangements of patients with BCM. In addition, we have demonstrated that the Mollon-Reffin (MR) Minimal test is a useful colour-discrimination test to aid in the diagnosis of BCM. Affected males were shown to fail the protan and deutan axes, but retained good discrimination on the tritan axis of the MR test, a compelling evidence for residual colour vision in BCM. This residual tritan discrimination was also readily detected with HRR plates. In two families, psychophysical testing demonstrated evidence for progression of disease. In two pedigrees, BCM could be linked to unequal crossovers within the opsin gene array that resulted in a single 5'-L/M-3' hybrid gene, with an inactivating Cys203Arg mutation. The causative mutations were not identified in the third family. CONCLUSIONS: The MR test is a useful method of detecting BCM across a wide range of age groups; residual tritan colour discrimination is clearly demonstrated and allows BCM to be distinguished from rod monochromatism. BCM is usually classified as a stationary cone dysfunction syndrome; however, two of our families show evidence of progression. This is the first report of progression associated with a genotype consisting of a single 5'-L/M-3' hybrid gene carrying an inactivating mutation. We have confirmed that the Cys203Arg inactivating mutation is a common sequence change in blue cone monochromats.
Adolescent↗
Immunological characterization of novel secreted antigens of Mycobacterium tuberculosis.
Proteins secreted by Mycobacterium tuberculosis are targets of host immune responses and as such are investigated for vaccine and immunodiagnostics development. Computer-driven searches of the M. tuberculosis H37Rv genome had previously identified 45 novel secreted proteins. Here, we report the characterization of these antigens in terms of specificity for the M. tuberculosis complex and the ability to induce human immune responses. BLAST homology searches and Southern hybridization identified 10 genes that were either specific for the M. tuberculosis complex or found in only two nontuberculous mycobacterial species of minor medical significance. Selected recombinant proteins were purified from Escherichia coli cells and tested for the ability to elicit antibody responses in tuberculosis patients. Reactivity of the serum panel was ' 36% with at least one of five novel proteins (Rv0203, Rv0603, Rv1271c, Rv1804c and Rv2253), 56% with the 38 kDa lipoprotein, a M. tuberculosis antigen known to be highly seroreactive, and 68% with a combination of Rv0203, Rv1271c and the 38 kDa antigen. Thus, at least five novel secreted proteins induce antibody responses during active disease; some of these proteins may increase the sensitivity of serological assays based on the 38 kDa antigen.
Antibodies, Bacterial↗
Comparison of flow cytometric methods for the measurement of ZAP-70 expression in a routine diagnostic laboratory.
Chronic lymphocytic leukaemia (CLL) follows a variable clinical course with patient survival ranging from only a few years despite treatment, to several decades in patients who may never require clinical intervention. Determination of the mutational status of a patient's immunoglobulin heavy chain variable region (Ig V(H)) gene has been used to provide prognostic information, but this assay is not available in most laboratories. The discovery of the expression of the protein tyrosine kinase zeta-associated protein (ZAP)-70 in V(H)-unmutated CLL cases led to its proposal as a surrogate marker for V(H) status. This study investigated the measurement of ZAP-70 expression in CLL using different flow cytometric protocols. Two different antibodies and two different staining methods were compared. The Caltag ZAP-70 antibody and Fix & Perm kit were the easiest to use and were the most sensitive and specific combination, with 91% concordance between ZAP-70 expression and V(H) status. Three patients (9%) were discordant (two V(H) mutated/ZAP-70 positive, and one V(H) unmutated/ZAP-70 negative). No correlation existed between CD38 and either ZAP-70 expression or V(H) status. Measurement of ZAP-70 expression using the Caltag antibody/kit combination provides a standardized flow cytometric method that could be introduced into a routine CLL immunophenotyping panel in a clinical diagnostic laboratory.
Aged↗
Pericardial infection caused by Brevibacterium casei.
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Actinomycetales Infections↗
Gibberellin inhibitors improve embryogenic tissue initiation in conifers.
Somatic embryogenesis (SE), the most promising technology to multiply high-value coniferous trees from advanced breeding and genetic engineering programs, is expected to play an important role in increasing productivity, sustainability, and uniformity of future forests in the United States. For commercial use, SE technology must work with a variety of genetically diverse trees. Initiation in loblolly pine (LP; Pinus taeda L.), our main focus species, is often recalcitrant for desirable genotypes. Initiation of LP, slash pine (SP; Pinus elliottii), Douglas-fir (DF; Pseudotsuga menziesii), and Norway spruce (NS; Picea abies) were improved through the use of paclobutrazol, a gibberellin synthesis inhibitor. Paclobutrazol was effective at concentrations ranging from 0.25 mg/l to 3.0 mg/l (0.85-10.2 microM) and optimal in LP at 1.0 mg/l. Using control media (no paclobutrazol) and 0.33-1.0 mg/l paclobutrazol, initiation percentages in LP, SP, DF, and NS were improved from 37.7% to 44.2% (across experiments), 19.3% to 28.5%, 16.9% to 23.7%, and 38.8% to 48.5%, respectively. Other gibberellin inhibitors such as flurprimidol, chlormequat-Cl, and daminozide also caused statistically significant increases in LP initiation when added to the medium at concentrations of 0.34, 10.0, and 1.0 microM, respectively. No detrimental effects on subsequent embryo development were observed when 29 new initiations from medium without GA inhibitor and 28 new initiations from medium containing paclobutrazol were tracked through culture capture, liquid culture establishment, cotyledonary embryo development, and germination.
Culture Techniques↗
Guidelines on the diagnosis and management of chronic lymphocytic leukaemia.
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Antineoplastic Agents↗
Achromatopsia caused by novel mutations in both CNGA3 and CNGB3.
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Adolescent↗
Inhaled benzo(a)pyrene impairs long-term potentiation in the F1 generation rat dentate gyrus.
The purpose of this study is to provide a point of reference regarding the neurotoxic effects resulting from exposure to environmental contaminants. Benzo(a)pyrene is a member of the polycyclic aromatic hydrocarbon (PAH) family and it is a by-product of combustion processes. Thus, persons living near factories or hazardous waste sites face the danger of exposure through contact with contaminated air, water and soil. In an effort to understand the impact of environmental contaminants, we have investigated the effects of gestational B(a)P aerosol exposure on long-term potentiation (LTP), a cellular correlate of learning and memory in the F1 generation. Briefly, timed-pregnant rats were exposed to B(a)P via nose-only inhalation on gestation days 11-21 for 4 hr per day. Dams were maintained to term and pups were weaned on postnatal day 30. Subsequent electrophysiological studies during postnatal days 60-70 revealed a diminution in LTP across the perforant path-granular cells synapses in the hippocampus of F1 generation animals that were transplacentally exposed to B(a)P aerosol relative to unexposed controls. Additionally, NMDA receptor subunit 1 (NR1) protein was found to be downregulated in the hippocampus of B(a)P exposed F1 generation animals. Taken together, our results suggest that gestational exposure to B(a)P aerosol attenuates the capacity for LTP in the F1 generation.
Administration, Inhalation↗
Evaluation of synthetic, M type-specific peptides as antigens in a multivalent group A streptococcal vaccine.
The recent development of emm gene sequence-based typing methodology has allowed group A streptococci (GAS) M serotype prevalence data to be determined. This information has been used to identify the components of a multivalent M protein peptide vaccine that could theoretically prevent most of the GAS-mediated diseases in the USA. In this study, we have evaluated in mice the immunogenicity and protective ability of multiple synthetic, M type-specific peptides, derived from the N-termini of three prevalent GAS serotypes (three peptides per serotype, total of nine peptides). At least one peptide, representing each of the three M types tested, was immunogenic. Five of the nine synthetic peptides tested, elicited an immune response in mice, and sera raised against four of the peptides, all possessed functional activity as demonstrated in a bactericidal assay. In vivo nasopharyngeal challenge experiments were carried out with peptides from the M1 (peptide M1-3) and M3 (peptide M3-2) proteins induced in vivo immune protection by reducing intranasal carriage. Reduction in colonization for M1-3 and M3-2 was 90% (P=0.02) and 66% (P<0.17), respectively. A reduction in colonization of 67% (P=0.03) was observed for M3-2 immunized mice when M43, a heterologous serotype, was used as the challenge strain. These results show the utility of synthetic, M type-specific peptides as antigens in a multivalent GAS vaccine.
Administration, Intranasal↗
The impact of Gelrite and activated carbon on the elemental composition of two conifer embryogenic tissue initiation media.
The elemental composition of plant tissue culture media was studied in response to (1) different levels of Gelrite and activated carbon (AC) in semisolid media and (2) different levels and types of AC in liquid media. Doubling the level of Gelrite from 2 g/l to 4 g/l reduced available magnesium (20%), calcium (16%), zinc (17%) and manganese (24%) and increased potassium (6%). AC adsorbed copper (90-95%) and zinc (35-51%) from both liquid and semisolid media. Two significantly different ACs gave minor differences in adsorption. No adsorption was indicated for inorganic anions. Nonacid-washed AC released significant levels of magnesium (44% increase), calcium (16% increase) and silica (a 75% increase to 1.8 mg/l). The elemental composition of media may need to be adjusted when increasing the Gelrite level or adding AC.
Adsorption↗
Improving loblolly pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology, germination, and gene expression.
Clonal production of loblolly pine ( Pinus taeda L.) through somatic embryogenesis has the potential to meet the increasing industrial demands for high-quality uniform raw materials. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Twenty-five newly initiated loblolly pine genotypes were followed through the process of liquid culture establishment, embryo maturation, germination, and retrieval from cryogenic storage. A maturation medium, capable of promoting the development of loblolly pine somatic embryos that can germinate, is presented that combines 1/2 P6 modified salts, 2% maltose, 13% polyethylene glycol 8000 (PEG), 5 mg/l abscisic acid (ABA), and 2.5 g/l Gelrite. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also described. A set of somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed. The quality of the resulting embryos was examined and compared to that of zygotic embryos using such parameters as morphology, dry weight, germination performance, and gene expression. All of the observations that were made support the conclusion that even with the new maturation medium somatic embryos grow approximately only halfway through the normal sequence of development and then prematurely discontinue growth.
Abscisic Acid↗