Evaluation of blood gene expression levels in facioscapulohumeral muscular dystrophy patients.

Facioscapulohumeral muscular dystrophy (FSHD) is caused by the expression of DUX4 in skeletal muscles. A number of therapeutic approaches are being developed to antagonize the events preceding and following DUX4 expression that leads to muscular dystrophy. Currently, the possibility to evaluate treatment response in clinical trials is hampered by the lack of objective molecular biomarkers connecting the disease cause to clinical performance. In this study we employed RNA-seq to examine gene expression in PAXgene tubes obtained from two independent cohorts of FSHD patients. Analysis of gene expression profiles did not lead to the identification of genes or pathways differentially expressed in FSHD patients, or associated with disease severity. In particular, we did not find evidence that the DUX4 and PAX7 signatures were differentially expressed. On the other hand, we were able to improve patient classification by including single genes or groups of genes in classification models. The best classifier was ROPN1L, a gene known to be expressed in testis, coincidentally the typical location of DUX4 expression. These improvements in patient classification hold the potential to enrich the FSHD clinical trial toolbox.

Informing a realistic laboratory erosion-testing regime--observations.

OBJECTIVES: To measure aspects of fizzy drink consumption in a social environment to inform the development of a laboratory testing regime. METHODS: This was an observational study in which participants were invited to attend one of four pizza and soft drink parties. All such foods and drinks were served in an air conditioned room at a temperature of 24°C. All drinks were at a temperature of 4°C and each participant was asked to spit out, into graduated cups, their first and second sips. Both the temperature and volume of these were measured. Upon completion of the party the volume of drinks consumed was determined. Video footage of the experiment was recorded for subsequent analysis to determine sip count and the elapsed time period between first and last sip. These values were compared to the analogous measured value of expectorated volume in order to assess the usefulness of video observation in the context of this work. RESULTS: The mean expectorated beverage temperature was 14.9±2.0°C. The mean time spent drinking was 44.2±17.4 minutes with a mean consumption rate of 13.3±6.0 ml/min. Only the sip volume and sip count per can values were significantly different between sexes (P<0.05) with females displaying lower values for volume and a higher sip count. There was close agreement between the sip volume values observed and calculated using video observation derived parameters. CONCLUSIONS: Several human drinking behaviour values were reported in this study and these will be of value in the development of more realistic laboratory erosion-testing regimes. It is concluded, within the limitations of this work, that (1) there are differences in the drinking behaviour of males and females with respect to sip volume and count, (2) the intraoral rise in temperature of a 4°C beverage is lower than that used in previous laboratory simulations and (3) the values derived from video observation agree with those measured directly validating this technique for use in further studies. CLINICAL SIGNIFICANCE: The work provides valuable quantitative data on which to base simulated laboratory erosion work. Perhaps the most significant finding is that sipped beverages attain a temperature of only 14.9°C.

Voluntary and reflex control of the human temporalis muscle.

Electromyographic recordings (EMGs) were made in 10 human subjects from the anterior and posterior parts of the temporalis muscle using skin surface electrodes. The activities produced by voluntary maximal clenching tasks and the reflex responses to electrical stimulation of the muco-gingival junction were studied. In most subjects, maximum activity in both parts of the muscle occurred when clenching in the intercuspal position (anterior temporalis: 7 of 10 subjects; posterior temporalis: 9 of 10 subjects). Clenching maximally in the retruded position usually resulted in less activity; when this activity was expressed as a percentage of the maximum achieved by each subject for that part of the muscle, the median values were: anterior temporalis, 68% and posterior temporalis, 79%. Clenching in the protruded position produced little or no activity (median values: anterior temporalis, 3%; posterior temporalis, 5%). There were no significant differences between the EMG activities of the anterior and posterior parts of the muscle during these tasks when the activities were normalized to the maximum achieved in each part of the muscle. Application of electrical stimuli at the muco-gingival junction (upper incisor region) produced reflex inhibitions and excitations in both parts of the muscle. There were no significant differences in the thresholds of these reflexes between the anterior and posterior parts of the muscle. Furthermore, there was little difference between the two parts of the muscle in terms of the latencies, durations and magnitudes of the responses. Thus the results of the study suggest that there are similar neural control mechanisms for the anterior and posterior parts of the temporalis muscle despite the common view that these parts of the muscle have different functions.

Bullae, bronchiectasis and nutritional emphysema in severe anorexia nervosa.

STUDY OBJECTIVES: Pulmonary complications of anorexia nervosa are rarely documented. The case of a patient with anorexia nervosa and pulmonary disease is presented, a new quantitative computed tomography (CT) method for the detection of emphysema is employed, the literature is reviewed and the concept of 'nutritional' emphysema is discussed. RESULTS: The case of a 34-year-old, nonsmoking woman with long-standing severe anorexia nervosa who was evaluated for cough and progressive shortness of breath is reported. Pulmonary function testing showed a predominant restrictive pattern with a marked reduction in carbon monoxide transfer and respiratory muscle strength, and an elevated residual volume. Imaging revealed bullae and bronchiectasis, and quantitative analysis of the CT scan was consistent with mild, generalized emphysema. Bronchial washings grew Pseudomonas aeruginosa. Known causes for bronchiectasis were excluded. A literature review disclosed few reported noninfectious pulmonary complications of anorexia nervosa. CONCLUSIONS: To the authors' knowledge, this is the first report of bullae and bronchiectasis in a patient with anorexia nervosa, and the CT analysis was consistent with mild emphysema. Malnutrition has been associated with emphysematous changes in animals and may be the primary insult in the development of emphysema, bullae and bronchiectasis in the present patient.

Attempted evaluation of three porcelain repair systems--what are we really testing?

Porcelain restorations are widely used in restorative dentistry with general success. On occasion however, fracture of the porcelain occurs. Sometimes a repair may be attempted using resin composite and one of the many bonding systems available. This in vitro investigation sought to determine and compare the shear bond strengths between a visible light-cured resin composite (Prisma TPH) and dental porcelain (Matchmaker Porcelain) for three different bonding systems (Scotchbond Multipurpose Plus (SMP); One-Step (OS); DenTASTIC (D)) with a view to conducting a larger study to assist in material selection. All measurements were performed one week following the preparation and storage of specimens in distilled water, at 37 degrees C, using a Universal Testing Machine at a crosshead speed of 0.5 mm/min. Although the mean shear bond strengths differed significantly (P < 0.05) (OS > SMP) and the Weibull modulus value of SMP was significantly (P < 0.05) lower than both OS and D the almost exclusive cohesive mode of failure observed, within the porcelain itself, suggested that the test was more a reflection of the quality of the underlying porcelain than the union under investigation. It was thus concluded that the shear bond strength test employed here, and in other studies did not truly reflect the shear bond strength at the porcelain/composite interface and that a more meaningful test should be developed. The observed cohesive failure may account for repeated clinical failures of porcelain repairs where bonding to an inherently flawed porcelain structure inevitably results in further failure.

Development, application and effectiveness of a novel logbook checklist assessment scheme in conservative dentistry.

In response to a lack of staff consistency in assessing clinical work, as identified by a student questionnaire, a checklist scheme of assessment was devised. This consisted of a series of questions covering the key stages of the majority of procedures in conservative dentistry to which a demonstrator indicates whether each aspect has been completed satisfactorily or not (yes/no) giving written comment to enhance the level of immediate feedback. Some 12 months following the introduction of the new logbook, a questionnaire was distributed to all 42 students who had experienced both this system and a previous ABC grading scale for completed work. This consisted of a series of 14 statements and sought to compare, for both systems, the clarity of assessment criteria, the relative level of staff consistency in applying these, effectiveness of feedback and overall meaningfulness of the grades awarded. In addition, in relation to self appraisal of performance, both the relevance of this to the practice of dentistry together with the degree the logbook facilitated this were assessed. The questionnaire return rate was 100%. Compared to the former grading system, the new logbook gave a significantly (P < 0.001) more meaningful measure of performance, indicated better what was being assessed (P < 0.001) and gave better feedback of those points requiring attention to improve performance. In addition, the consistency of staff applying the checklist had improved significantly (P < 0.05) since the introduction of the logbook. It was thus concluded that the logbook was a versatile tool in assessment of the clinical skills in conservative dentistry.

The reinnervation of healing extraction sockets in the ferret.

The objective of this study was to describe the pattern of healing of pulpal and periodontal nerve fibers following tooth extraction. The mandibular canine teeth were, under general anesthesia, removed from one side of nine young ferrets. One week (two animals), one month (four animals), and three months (three animals) later, again under general anesthesia, the animals were perfused with fixative. The mandibles were decalcified, paraffin-embedded, and serially sectioned. Sample sections were stained with hematoxylin and eosin, the remainder with a silver stain for nerves. Serial reconstructions were made of the innervation in control teeth and in the extraction sockets. A grid sampling technique was used for the estimation of the innervation density in different regions at various levels in the healing socket. One week after extraction, although osteoid material was beginning to fill the socket, the pattern of innervation remained similar to that of controls in being restricted to the former location of the periodontal ligament and being densest in the apical third of the socket. One month after extraction, the innervation consisted of many fine axons or small bundles distributed throughout the healing osteoid tissue though still most dense in the periphery of the socket. The overall innervation density increased between one week and one month post-extraction. At three months, as the margins of the socket were becoming obscured, the innervation consisted largely of one or two organized nerve trunks running from the apical to the coronal aspect in either the central or lateral aspects of the socket. At no time was there evidence of neuroma formation.