A systematic review evaluating the use of ultrasound in the identification of osteochondrosis in horses.

Osteochondrosis is commonly encountered in young horses, with welfare, performance, and economic effects. Consequently, pre-purchase radiographic screening for osteochondrosis is routinely performed. Ultrasonographic examination of articular cartilage and osteochondrosis lesions are described in the literature with many case series or single case reports published. This systematic review was undertaken to examine the evidence for using ultrasonography in comparison to traditional radiography, arthroscopy or necropsy findings in the detection of osteochondrosis. The systematic review identified a paucity of studies in which there was marked variation in the populations, sample size, methods and results reported. Currently, there is no strong evidence confirming the diagnostic accuracy and validity of ultrasonography in the detection of osteochondral lesions in the relevant joints in horses.

Profile and management of acute coronary syndromes at primary- and secondary-level healthcare facilities in Cape Town.

BACKGROUND: Little is known about the clinical profile and management of patients with acute coronary syndromes (ACS) in the South African public sector. METHODS: We conducted a retrospective study of patients presenting with ACS to a secondary-level healthcare facility in Cape Town during a one-year period to study the clinical profile and management of these patients. RESULTS: Among the 214 patients in this cohort, 48 (27.5%) had ST-segment elevation myocardial infarction (STEMI), 43 (24.7%) had non-ST-segment elevation myocardial infarction and 83 (47.7%) unstable angina pectoris. We identified high rates of >12-hour delays in first medical contact after symptom onset (46%) and inaccurate ECG diagnosis of STEMI (29.2%), which were associated with low rates of thrombolysis (39.6%). High rates of non-adherence and ACS recurrence were also observed. CONCLUSION: To address the local challenges in ACS management highlighted in this study, we propose the development of a regional referral network prioritising access to expedited care and primary reperfusion interventions in ACS.

Role of the virtual reality simulator (ScanTrainer) as a multidisciplinary training tool in transvaginal ultrasound: A systematic review and narrative synthesis.

OBJECTIVES: This systematic review investigates the role of the ScanTrainer as a virtual reality training simulator and its impact on the scanning skills in transvaginal ultrasound of novice ultrasound practitioners. KEY FINDINGS: After searching ten databases for studies incorporating the simulator as a part of the learning/training process, ten out of 684 studies met the eligibility criteria and were included in the review. The analysis of the textual findings using narrative synthesis approach resulted in four themes: Validation (assessment of the validity of the simulator), Learning (using the simulator as a learning tool), Perspectives (the perceptions of participants trained on the simulator), and Transferable skills (skills developed on the simulator can transfer to clinical practice). CONCLUSION: Although literature indicates that the simulator is valuable as a training/learning tool, there is insufficient evidence of measurable effects on clinical practice of simulator usage by different healthcare professions.

Centre-level variation in treatment and outcomes and predictors of outcomes in 5-year-old children with non-syndromic unilateral cleft lip treated within a centralized service: the Cleft Care UK study. Part 6: summary and implications.

OBJECTIVES: To summarize and discuss centre-level variation across a range of treatment and outcome measures and examine individual and ecological determinants of outcome in children in Cleft Care UK (CCUK). SETTING AND SAMPLE POPULATION: Two hundred and sixty-eight 5-year-old British children with non-syndromic unilateral cleft lip and palate (UCLP) recruited to CCUK and treated within a centralized service. MATERIALS AND METHODS: Children had a range of treatment and outcome measures collected at a 5-year audit clinic. These outcomes included dento-alveolar arch relationships from study models, measures of facial appearance from cropped photographs, hearing loss from audiological assessment, speech from speech recordings, self-confidence and strengths and difficulties from parental self-report. Data were collected on educational attainment at age 7 using record linkage. Centre variation was examined using hierarchical regression and associations between variables were examined using logistic or poisson regression. RESULTS: There was centre-level variation for some treatments (early grommet placement, fitting of hearing aids, fluoride treatment, secondary speech surgery and treatment for cleft speech characteristics) and for some outcomes (intelligibility of speech). Hearing loss was associated with a higher risk of poor speech while speech therapy was associated with a lower risk of poor speech. Children had high levels of caries but levels of preventative treatment (fluoride varnish and tablets) were low. CONCLUSIONS: Further improvements to and monitoring of the current centralized model of care are required to ensure the best outcomes for all children with cleft lip and palate.

Neuropathic pain prevalence following spinal cord injury: A systematic review and meta-analysis.

Following spinal cord injury (SCI), chronic pain is a common secondary complication with neuropathic pain (NP) cited as one of the most distressing and debilitating conditions leading to poor quality of life, depression and sleep disturbances. Neuropathic pain presenting at or below the level of injury is largely refractory to current pharmacological and physical treatments. No consensus on the prevalence of NP post SCI currently exists, hence this systematic review was undertaken. The review comprised three phases: a methodological assessment of databases [PubMed, Embase, Web of Knowledge, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Cochrane Library and Physiotherapy Evidence Database (PEDro)] identifying potential papers and screening for inclusion criteria by two independent reviewers; data extraction; and finally rating of internal validity and strength of the evidence, using a published valid and reliable scale. Meta-analysis estimated pooled point prevalence rates using a random effects model. In total, 17 studies involving 2529 patients were included in the review. Overall point prevalence rates for NP were established at 53% (38.58-67.47); 19% (13.26-26.39) for at-level NP and 27% (19.89-34.61) for below-level NP, with high heterogeneity noted (I2  = 84-93%). Prevalence rates for NP following SCI are high. Future studies should include established definitions, classification systems and assessment tools for NP at defined time points post SCI to follow the trajectory of this problem across the lifespan and include indices of sleep, mood and interference to allow for appropriate, optimal and timely NP management for each patient. WHAT DOES THIS REVIEW ADD?: This is the first systematic review and meta-analysis to record pooled point prevalence of neuropathic pain post spinal cord injury at 53%. Additional pooled analysis shows that neuropathic pain is more common below the level of lesion, in patients with tetraplegia, older patients and at 1 year post injury.

Spatial and temporal analysis of litter in the Celtic Sea from Groundfish Survey data: Lessons for monitoring.

The Marine Strategy Framework Directive requires EU Member States to sample and monitor marine litter. Criteria for sampling and detecting spatial and/or temporal variation in the amount of litter present have been developed and initiated throughout Europe. These include implementing standardised sampling and recording methods to enable cross-comparison and consistency between neighbours. Parameters of interest include; litter occurrence, composition, distribution and source. This paper highlights the litter-related initiatives occurring in Irish waters; presents an offshore benthic litter sampling series; provides a power analysis to determine trend detection thresholds; identifies areas and sources of litter; and proposes improvements to meet reporting obligations. Litter was found to be distributed throughout Irish waters with highest occurrences in the Celtic Sea. Over 50% of litter encountered was attributed to fishing activities: however only a small proportion of the variability in litter occurrence could be explained by spatial patterns in fishing effort. Issues in implementing standardised protocol were observed and addressed.

Sparse and incomplete factorial matrices to screen membrane protein 2D crystallization.

Electron crystallography is well suited for studying the structure of membrane proteins in their native lipid bilayer environment. This technique relies on electron cryomicroscopy of two-dimensional (2D) crystals, grown generally by reconstitution of purified membrane proteins into proteoliposomes under conditions favoring the formation of well-ordered lattices. Growing these crystals presents one of the major hurdles in the application of this technique. To identify conditions favoring crystallization a wide range of factors that can lead to a vast matrix of possible reagent combinations must be screened. However, in 2D crystallization these factors have traditionally been surveyed in a relatively limited fashion. To address this problem we carried out a detailed analysis of published 2D crystallization conditions for 12 ß-barrel and 138 α-helical membrane proteins. From this analysis we identified the most successful conditions and applied them in the design of new sparse and incomplete factorial matrices to screen membrane protein 2D crystallization. Using these matrices we have run 19 crystallization screens for 16 different membrane proteins totaling over 1300 individual crystallization conditions. Six membrane proteins have yielded diffracting 2D crystals suitable for structure determination, indicating that these new matrices show promise to accelerate the success rate of membrane protein 2D crystallization.

Advantages of indium-tin oxide-coated glass slides in correlative scanning electron microscopy applications of uncoated cultured cells.

A method of direct visualization by correlative scanning electron microscopy (SEM) and fluorescence light microscopy of cell structures of tissue cultured cells grown on conductive glass slides is described. We show that by growing cells on indium-tin oxide (ITO)-coated glass slides, secondary electron (SE) and backscatter electron (BSE) images of uncoated cells can be obtained in high-vacuum SEM without charging artefacts. Interestingly, we observed that BSE imaging is influenced by both accelerating voltage and ITO coating thickness. By combining SE and BSE imaging with fluorescence light microscopy imaging, we were able to reveal detailed features of actin cytoskeletal and mitochondrial structures in mouse embryonic fibroblasts. We propose that the application of ITO glass as a substrate for cell culture can easily be extended and offers new opportunities for correlative light and electron microscopy studies of adherently growing cells.

Pre prosthetic mobility: the amputees' perspectives.

PURPOSE: It is considered good practice that amputees remain in a wheelchair until fitted with a prosthesis. However, this practice is not evidence based. In the first of a series of studies in pre prosthetic mobility, this study will explore the individual amputees' perspective. METHOD: A total of 25 participants from a regional disablement service centre were interviewed focusing on their experiences of getting around the home post discharge. The transcripts were coded for types of mobility methods used and the content analysed. RESULTS: Whilst the wheelchair was the predominant method of mobilizing, this was not viewed positively by amputees. Crutches were frequently used, even if crutches were not provided by therapists. Participants cited space and choice as reasons behind this, in particular the psychological impact of being 'stuck' in a wheelchair. Other methods cited included crawling and bottom shuffling. CONCLUSION: Amputees do not always comply with therapists' advice and frequently use other methods of mobilizing as a matter of choice as well as necessity. The risk factors associated with these methods is a priority for future research.

Improvements to a cryosystem to observe ice nucleating in a variable pressure scanning electron microscope.

The variable pressure scanning electron microscope (VPSEM) has expanded the scope of the SEM to allow the imaging of dynamic, electrically insulating systems. The use of water vapor as the imaging gas present in the chamber allows the successful imaging of hydrated samples. As awareness of the system capabilities becomes more well known, greater pressure has been put onto the microscopist to push the boundaries of both temperature and resolution for the study of diverse hydrated samples whose dynamics may not occur at the usual room temperatures in a VPSEM. In this article we discuss the stages in the development of a cryosystem that has led to the successful observation of the nucleation of ice from a solution in situ. This investigation also leads to further possibilities of imaging hydrated samples in the little explored temperature range of 188-238 K (from -85 to-35 degrees C). This study includes the exploration of how the temperature of various surfaces inside the microscope will change the system's ability to keep a sample hydrated or in its native state.

Modulation of TGF-beta signaling by proinflammatory cytokines in articular chondrocytes.

OBJECTIVE: The normal structure and function of articular cartilage are the result of a precisely balanced interaction between anabolic and catabolic processes. The transforming growth factor-beta (TGF-beta) family of growth factors generally exerts an anabolic or repair response; in contrast, proinflammatory cytokines such as interleukin 1 beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) exert a strong catabolic effect. Recent evidence has shown that IL-1beta, and TNF-alpha, and the TGF-beta signaling pathways share an antagonistic relationship. The aim of this study was to determine whether the modulation of the response of articular chondrocytes to TGF-beta by IL-1beta or TNF-alpha signaling pathways occurs through regulation of activity and availability of mothers against DPP (Drosophila) human homologue (Smad) proteins. METHODS: Human articular chondrocytes isolated from knee joints from patients with osteoarthritis (OA) or normal bovine chondrocytes were cultured in suspension in poly-(2-hydroxyethyl methacrylate)-coated dishes with either 10% fetal bovine serum media or serum-deprived media 6h before treatment with IL-1beta alone, TNF-alpha alone or IL-1beta followed by TGF-beta. Nuclear extracts were examined by electrophoretic mobility-shift assays (EMSA) for nuclear factor-kappa B (NF-kappaB) and Smad3/4 deoxyribonucleic acid (DNA) binding. Nuclear extracts were also subjected to the TranSignal Protein/DNA array (Panomics, Redwood City, CA) enabling the simultaneous semiquantitative assessment of DNA-binding activity of 54 different transcription factors. Nuclear phospho-Smad2/3 and total Smad7 protein expression in whole cell lysates were studied by Western blot. Cytoplasmic Smad7, type II collagen alpha 1 (COL2A1), aggrecan and SRY-related high mobility group-Box gene 9 (SOX-9) mRNA expression were measured by real-time polymerase chain reaction (PCR). RESULTS: The DNA-binding activity of Smad3/4 in the TranSignal Protein/DNA array was downregulated by TNF-alpha (46%) or IL-1beta treatment (42%). EMSA analysis showed a consistent reduction in Smad3/4 DNA-binding activity in human articular chondrocytes treated with IL-1beta or TNF-alpha. TGF-beta-induced Smad3/4 DNA-binding activity and Smad2/3 phosphorylation were also reduced following pretreatment with IL-1beta in human OA and bovine chondrocytes. Real-time PCR and Western blot analysis showed that IL-1beta partially reversed the TGF-beta stimulation of Smad7 mRNA and protein levels in TGF-beta-treated human OA cells. In contrast, TGF-beta-stimulated COL2A1, aggrecan, and SOX-9 mRNA levels were abrogated by IL-1beta. CONCLUSIONS: IL-1beta or TNF-alpha exerted a suppressive effect on Smad3/4 DNA-binding activity in human articular chondrocytes, as well as on TGF-beta-induced stimulation of Smad3/4 DNA-binding activity and Smad2/3 phosphorylation in human OA and bovine articular chondrocytes. IL-1beta partially reversed the increase in TGF-beta-stimulated Smad7 mRNA or protein levels suggesting that Smad7 may not be involved in the suppression of TGF-beta signaling induced by IL-1beta or TNF-alpha in articular chondrocytes. The balance between the IL-1beta or TNF-alpha and the TGF-beta signaling pathways is crucial for maintenance of articular cartilage homeostasis and its disruption likely plays a substantial role in the pathogenesis of OA.

A technique for improved focused ion beam milling of cryo-prepared life science specimens.

The combination of focused ion beam and scanning electron microscopy with a cryo-preparation/transfer system allows specimens to be milled at low temperatures. However, for biological specimens in particular, the quality of results is strongly dependent on correct preparation of the specimen surface. We demonstrate a method for deposition of a protective, planarizing surface layer onto a cryo-sample, enabling high-quality cross-sectioning using the ion beam and investigation of structures at the nanoscale.

Synthesis and characterization of SERS gene probe for BRCA-1 (breast cancer).

A protocol for binding cresyl fast violet (CFV), a SERS-active dye (label) containing an aromatic amino group with a modified oligomer having a carboxy derivatized thymidine moiety using carbodiimide coupling has been achieved for the first time. Covalent coupling between CFV and the oligomer has been confirmed by mass spectral analysis of the labeled oligomer. The fluorescence, SERS and absorption characteristics of the labeled product have been evaluated. The chosen oligomer contains a BRCA-1 (breast cancer) sequence, and hence has the potential for being used as a gene probe to identify BRCA-1 gene. It has high potential for being used in polymerase chain reaction (PCR) amplification, as has been performed with labeled oligonucleotide for the HIV sequence.

Dealing with lead in Broken Hill--trends in blood lead levels in young children 1991-2003.

The objective of the study was to investigate trends in blood lead concentrations in preschool children between 1991 and 2003, as part of the evaluation strategy of a public health lead management program in Broken Hill, Australia. Since 1991, all Broken Hill children aged 1-4 years have been offered at least annual blood lead screening as part of a community-wide lead management program. Recruitment of children was promoted throughout the period using local media and distribution of promotional material from health care centres and preschool, childcare, and educational facilities around the city. Venous blood samples were collected using standard procedures and analyses were subjected to internal and external quality control programs. Because the frequency distribution of blood lead levels are skewed, geometric rather than arithmetic means were used for comparative purposes. Trend analysis was based on age and sex standardised mean blood lead levels. The number of 1- to 4-year-old children screened ranged between 496 and 948 in any one year and response rates varied between 39% and 73%. The age-sex standardised mean blood lead level decreased from 16.3 microg/dL to 7.1 microg/dL between 1991 and 2003. Overall, blood lead levels declined by 56% over 13 years. These reductions were consistently observed irrespective of age or where a child lived in the town. The rate of decline has slowed since 1997. We conclude that substantial progress has been made in dealing with the lead problem in Broken Hill children, although the rate of decline of blood lead levels has slowed. Continued public health action is still needed to bring the proportion of young children with significantly elevated blood lead levels (>15 microg/dL) down from the 2003 figure of 12% to the NHMRC community-based target for lead in young Australians of 5%.

Morphological instability in InAs/GaSb superlattices due to interfacial bonds.

Synchrotron x-ray diffraction is used to compare the misfit strain and composition in a self-organized nanowire array in an InAs/GaSb superlattice with InSb interfacial bonds to a planar InAs/GaSb superlattice with GaAs interfacial bonds. It is found that the morphological instability that occurs in the nanowire array results from the large misfit strain that the InSb interfacial bonds have in the nanowire array. Based on this result, we propose that tailoring the type of interfacial bonds during the epitaxial growth of III-V semiconductor films provides a novel approach for producing the technologically important morphological instability in anomalously thin layers.

Correction of a mineralization defect by overexpression of a wild-type cDNA for COL1A1 in marrow stromal cells (MSCs) from a patient with osteogenesis imperfecta: a strategy for rescuing mutations that produce dominant-negative protein defects.

Gene therapy for dominant-negative disorders presents a more difficult challenge than gene therapy for recessive disorders, since even partial replacement of a protein for a recessive disorder can reverse symptoms. Osteogenesis imperfecta (OI) has frequently served as a model disorder for dominant-negative defects of structural proteins. The disease is caused by mutations in type I collagen (COL1A1), the major structural component of bone, skin and other connective tissues. The severity of the phenotype is largely dependent on the ratio of normal to mutant type I procollagen synthesized by cells. Recently, attempts have been made to develop strategies for cell and gene therapies using the adult stem cells from bone marrow referred to as mesenchymal stem cells or marrow stromal cells (MSCs). In this study, we used MSCs from a patient with type III OI who was heterozygous for an IVS 41A+4C mutation in COL1A1. A hybrid genomic / cDNA construct of COL1A1 was transfected into the MSCs and the transfectants were expanded over a 200-fold. Transfected MSCs showed increased expression of the wild-type mRNA and protein. In vitro assays demonstrated that the transfected cells more efficiently differentiated into mineralizing cells. The results indicated that it is possible to overexpress COL1A1 cDNA in OI MSCs and thereby to correct partially the dominant-negative protein defect.

The effect of burn injury on adolescents autobiographical memory.

Autobiographical memory recall was investigated in two female adolescent groups; one group who had experienced a burn injury and a matched control group. The Burn group was not currently depressed or anxious, but scored significantly higher on the intrusion subscale of the impact of event scale compared to controls. Two autobiographical memory tasks, the autobiographical memory cueing task and the Children's Autobiographical Memory Inventory (CAMI), were used. For the cueing task, the Burn group was significantly slower to recall specific memories. This group also recalled significantly fewer specific memories and significantly more extended overgeneral memories. For the CAMI, the burns group produced significantly lower semantic and episodic recall. The Burn group also produced significant correlations between sub-scales of the impact of event scale and selected measures on the autobiographical memory tasks. Higher intrusion scores were associated with less detailed episodic recall. Higher avoidance scores were associated with longer latencies to recall memories to negative cue words and fewer specific memories to all cue words. These results are discussed from the perspective that the Burn group experienced intrusive thoughts which interfered with normal autobiographical functioning.

The mechanics of calcium transport.

With the recent atomic models for the sarcoplasmic reticulum Ca(2+)-ATPase in the Ca(2+)-bound state, the Ca(2+)-free, thapsigargin-inhibited state, and the Ca(2+)-free, vanadate-inhibited state, we are that much closer to understanding and animating the Ca(2+)-transport cycle. These "snapshots" of the Ca(2+)-transport cycle reveal an impressive breadth and complexity of conformational change. The cytoplasmic domains undergo rigid-body movements that couple the energy of ATP to the transport of Ca2+ across the membrane. Large-scale rearrangements in the transmembrane domain suggest that the Ca(2+)-binding sites may alternately cease to exist and reform during the transport cycle. Of the three cytoplasmic domains, the actuator (A) domain undergoes the largest movement, namely a 110 degrees rotation normal to the membrane. This domain is linked to transmembrane segments M1-M3, which undergo large rearrangements in the membrane domain. Together, these movements are a main event in Ca2+ transport, yet their significance is poorly understood. Nonetheless, inhibition or modulation of Ca(2+)-ATPase activity appears to target these conformational changes. Thapsigargin is a high-affinity inhibitor that binds to the M3 helix near Phe256, and phospholamban is a modulator of Ca(2+)-ATPase activity that has been cross-linked to M2 and M4. The purpose of this review is to postulate roles for the A domain and M1-M3 in Ca2+ transport and inhibition.

Static and dynamic experiments in cryo-electron microscopy: comparative observations using high-vacuum, low-voltage and low-vacuum SEM.

We carried out a unique comparative study between three modes of cryo-scanning electron imaging: high-vacuum, low-voltage and low-vacuum, using ice cream as a model system. Specimens were investigated both with and without a conductive coating (Au/Pd) and at temperatures for which ice either remains fully frozen (< -110 degrees C) or undergoes sublimation (-110 to -90 degrees C). At high magnification, high-vacuum imaging of coated specimens gave the best results for 'static' specimens (i.e. containing fully frozen ice). Low voltages, such as 1 kV, could be used for imaging uncoated specimens at high vacuum, although slight 'classical' charging artefacts remained an issue, and the reduced electron beam penetration tended to decrease the definition between different microstructural features. However, this mode was useful for observing in situ sublimation from uncoated specimens. Low-vacuum mode, involving small partial pressures of nitrogen gas, was particularly suited to in situ sublimation work: when sublimation was carried out in low vacuum in the absence of an anti-contaminator plate, sublimation rates were significantly reduced. This is attributed to a small partial pressure of sublimated water vapour remaining near the specimen surface, enhancing thermodynamic stability.