To accept, or not to accept? A service evaluation to appraise complexity assessment of orthodontic patients referred into a secondary care setting.

Introduction Commissioners in England use the Commissioning for Quality and Innovation (CQUIN) payments mechanism to encourage the best provision of orthodontic treatment. However, CQUIN only use the patient's orthodontic need as a measure of complexity, rather than the levels outlined in the orthodontic commissioning guide published by NHS England. A service evaluation was designed to ascertain a secondary care setting's compliance with the commissioning complexity levels, as a new comparator for CQUIN case-mix assessment.Materials and methods A prospective evaluation was conducted for all new patients referred to the Mid Yorkshire NHS Trust orthodontic department in a 12-month period, using the levels categorised by the commissioning guide. A standard was set to accept no fewer than 80% level 3b patients.Results Of patients accepted for orthodontic treatment, 89.9% were of the highest level 3b complexity. This was compared to only 69.8% of patients having an Index of Orthodontic Treatment Need, Dental Health Component, 5.Conclusion The findings support a recommendation that commissioners should consider complexity based on the commissioning guidance, rather than orthodontic need alone; it is important that the economic drivers of commissioning implementation fairly reflect the specialist work being carried out by the workforce.

The first permanent molar: spontaneous eruption after a five-year failure.

BACKGROUND: It is rare for a first permanent molar (FPM) to temporarily exhibit clinical features of failure of eruption, followed by regeneration of full eruptive capacity 5 years later. Indeterminate failure of eruption (IFE) is a diagnosis of exclusion where the distinction between primary failure of eruption (PFE) and mechanical failure of eruption (MFE) is unclear, including patients too young to specify. CASE REPORT: An 11-year-old girl attended the orthodontic clinic at Mid Yorkshire Hospitals NHS Trust regarding an unerupted lower right FPM. Her medical and dental trauma history was unremarkable. She presented with a Class II division 2 malocclusion in the mixed dentition, with all other FPMs fully erupted. CONCLUSION: This report documents that an unerupted FPM in an 11-year-old patient may still have the eruptive potential to become functional within the dentition. The period spent monitoring the FPM's outcome prior to surgical intervention has avoided an operation under general anaesthetic and potentially unnecessary orthodontic treatment, as the tooth subsequently erupted without treatment.

Microbial contamination of "as received" and "clinic exposed" orthodontic materials.

INTRODUCTION: Our objective was to determine whether components of fixed orthodontic appliances as received from the manufacturers and after exposure to the clinical environment are free from microbial contamination before clinical use. A pilot molecular microbiologic laboratory study was undertaken at a dental hospital in the United Kingdom. METHODS: A range of orthodontic materials "as received" from the manufacturers and materials "exposed" to the clinical environment were studied for bacterial contamination. After growth on blood-rich media, cultured bacteria were identified by 16S rDNA polymerase chain reaction amplification and sequence phylogeny. RESULTS: Bacteria were isolated from "as received" bands, archwires, and impression trays, but the level of contamination was low (0.5 × 10(1) to 1.825 × 10(2) CFU/mL(-1)). Various bacterial species were isolated from "clinic exposed" bands, archwires, impression trays, coil springs, and elastomeric modules, but the level of contamination was low (0.5 × 10(1) to 8.0 × 10(1) CFU/mL(-1)). The most commonly identified bacterial species was Staphylococcus epidermidis, followed by Kocuria, Moraxella, and Micrococcus species. CONCLUSIONS: New materials "as received" from the manufacturers and those exposed to the clinical environment are not free from bacterial contamination before use in patients, but this contamination is low considering the potential for aerosol and operator contamination and could be considered insignificant. Further studies would be required to determine the level of risk that this poses.

Microarray analysis of embryonic stem cells and differentiated embryoid bodies.

By altering the cellular microenvironment and culture media composition, embryonic stem cells (ESCs) can be induced to differentiate in vitro into somatic cell types from the three primitive germ layers. ESC differentiation is regulated by an intricate series of signaling events that result in their transcriptional reprogramming, asymmetric cell division, and differentiation. Using various pharmacological agents and/or genetic manipulations, one can drive and enrich ESC differentiation to specific cell lineages. Identifying the transcriptional fingerprint during ESC differentiation could yield novel targets for genetic or pharmacological regulation to increase the yield of desirable cell types. We discuss here how to culture undifferentiated mouse ESCs (E14 line from 129/Ola) and generate embryoid bodies (EBs). We also discuss in detail how to prepare Affymetrix samples, how to hybridize and scan arrays, and how to quality control data and generate signal values and permutation based P-values.

Increased DNA microarray hybridization specificity using sscDNA targets.

BACKGROUND: The most widely used amplification method for microarray analysis of gene expression uses T7 RNA polymerase-driven in vitro transcription (IVT) to produce complementary RNA (cRNA) that can be hybridized to arrays. However, multiple rounds of amplification are required when assaying very small amounts of starting RNA. Moreover, certain cRNA-DNA mismatches are more stable than the analogous cDNA-DNA mismatches and this might increase non-specific hybridization. We sought to determine whether a recently developed linear isothermal amplification method (ribo-SPIA) that produces single stranded cDNA would offer advantages over traditional IVT-based methods for microarray-based analyses of transcript expression. RESULTS: A single round of ribo-SPIA amplification produced sufficient sscDNA for hybridizations when as little as 5 ng of starting total RNA was used. Comparisons of probe set signal intensities obtained from replicate amplifications showed consistently high correlations (r = 0.99). We compared gene expression in two different human RNA samples using ribo-SPIA. Compared with one round IVT, ribo-SPIA had a larger dynamic range and correlated better with quantitative PCR results even though we used 1000-fold less starting RNA. The improved dynamic range was associated with decreases in hybridization to mismatch control probes. CONCLUSION: The use of amplified sscDNA may offer substantial advantages over IVT-based amplification methods, especially when very limited amounts of starting RNA are available. The use of sscDNA targets instead of cRNA targets appears to improve hybridization specificity.

E2F-1 regulates the expression of a subset of target genes during skeletal myoblast hypertrophy.

Cellular hypertrophy, or growth without division, is an adaptive response to various physiological and pathological stimuli in postmitotic muscle. We demonstrated previously that angiotensin II stimulates hypertrophy in C2C12 myoblasts by transient activation of the cyclin-dependent kinase 4 complex, subsequent phosphorylation of retinoblastoma protein, release of histone deacetylase 1 from the retinoblastoma protein inhibitory complex, and partial activation of the transcription factor E2F-1. These observations led us to propose a model in which partial inactivation of the retinoblastoma protein complex leads to the derepression of a subset of E2F-1 targets necessary for cell growth without division during hypertrophy. We now present data that support this model and suggest the mechanism by which E2F-1 regulates hypertrophy. We examined expression profiles of angiotensin II-stimulated myoblasts and identified a subset of E2F-1 target genes that are specifically regulated during the hypertrophic response. We showed that the expression of E2F-1 targets involved in G1/S transit, DNA replication, and mitosis is not altered during the hypertrophic response, while the expression of E2F-1-regulated genes controlling early G1 progression, cytoskeletal organization, protein synthesis, mitochondrial function, and programmed cell death is up-regulated. Furthermore, we demonstrated that activation of cytochrome c oxidase genes occurs during the development of hypertrophy and that cytochrome c oxidase IV is a direct transcriptional target of E2F-1. These studies demonstrated that E2F-1 activity at specific promoters is dependent on physiological circumstances and that E2F-1 should be considered a potential target in the treatment of pathologic hypertrophy.

Gene expression profiles during human CD4+ T cell differentiation.

To develop a comprehensive catalogue of phenotypic and functional parameters of human CD4(+) T cell differentiation stages, we have performed microarray gene expression profiling on subpopulations of human thymocytes and circulating naive CD4(+) T cells, including CD3(-)CD4(+)CD8(-) intrathymic T progenitor cells, CD3(int)CD4(+)CD8(+) 'double positive' thymocytes, CD3(high)CD4(+)CD8(-) 'single positive' thymocytes, CD3(+)CD4(+)CD8(-) CD45RA(+)CD62L(+) naive T cells from cord blood and CD3(+)CD4(+)CD8(-) CD45RA(+)CD62L(+) naive T cells from adult blood. These subpopulations were sort-purified to >98% purity and their expressed RNAs were analyzed on Affymetrix Human Genome U133 arrays. Comparison of gene expression signals between these subpopulations and with early passage fetal thymic stromal cultures identify: (i) transcripts that are preferentially expressed in human CD4(+) T cell subpopulations and not in thymic stromal cells; (ii) major shifts in gene expression as progenitor T cells mature into progeny; (iii) preferential expression of transcripts at the progenitor cell stage with plausible relevance to the regulation of expansion and differentiation of these cells; and (iv) preferential expression of potential markers of recent thymic emigrants in naive-phenotype CD4(+) T cells from cord blood. Further evaluation of these findings may lead to a better definition of human thymopoiesis as well as to improved approaches to monitor and to augment the function of this important organ of T cell production.