Evaluation of the total adenylate (ATP + ADP + AMP) test for cleaning verification in healthcare settings.

INTRODUCTION: Evaluation of cleaning methods is the first step in the prevention of healthcare-associated infections. ATP hygiene monitoring tests are widely used for assessing the effectiveness of cleaning procedures. The test is easy to use and gives immediate results, however, ATP can be metabolized and degraded to ADP and AMP. Recently, a total adenylate [ATP + ADP + AMP(A3)] monitoring test has been developed. Our objective was to evaluate the usefulness of the A3 test for cleaning verification in healthcare settings. METHODS: The detection sensitivities of the ATP and the A3 tests were compared using blood, and debris derived from gloved-hand method and endoscopes immediately after endoscopic examination. The performance of the A3 test in monitoring cleanliness of high touch surfaces in the hospital and endoscopes at each cleaning step was also evaluated. RESULTS: For the hemolysate, the measurement values of the A3 test were stable, although ATP was promptly degraded. In debris from hands, the amount of A3 was 20 times higher than that of ATP. The detection sensitivities of the A3 test on residues derived from gastroscopes and colonoscopes were 3 and 8 times higher, respectively, than those from the ATP test. A field study indicated that a large number of microorganisms tend to show high A3 values on high touch surfaces in the hospital and on endoscopes. CONCLUSIONS: The A3 test showed higher detection sensitivities than the conventional ATP test for organic debris associated with healthcare settings.

Long-term side effects on the temporomandibular joints and oro-facial function in patients with obstructive sleep apnoea treated with a mandibular advancement device.

Patients with obstructive sleep apnoea (OSA) in long-term treatment with a mandibular advancement device (MAD) to increase the upper airway space may develop changes in the temporomandibular joint (TMJ) and the oro-facial function due to the protruded jaw position during sleep. The aim was to investigate the influence of long-term MAD treatment on the TMJs, oro-facial function and occlusion. This prospective study included 30 men and 13 women (median age 54) with OSA [Apnoea-Hypopnoea Index (AHI): 7-57]. They were examined with the Nordic Orofacial Test Screening (NOT-S), the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) and cone beam computed tomography (CBCT) of the TMJs. The examination was performed before MAD treatment (T0), and 3-6 months (T1, no CBCT), 1 year (T2) and 3 years (T3) after treatment start. The results were analysed as long term (T0-T3, n = 14) and short term (T0-T2, n = 24) by t-test, Fisher's exact test and anova. Both long- and short-term analyses revealed a reduction in AHI (P < 0·002). Significant long term were increased scores in the NOT-S Interview (P < 0·045), reduced vertical overbite (P < 0·031) and increased jaw protrusive movement (P < 0·027). TMJ changes were found as joint sounds in terms of reciprocal clicking and crepitus, short term as a decrease and subsequent recurrence (P < 0·053; P < 0·037). No significant radiological changes were found. In conclusion, MAD treatment is beneficial to some OSA patients, but might induce changes in the TMJs, the oro-facial function and the occlusion. However, these changes seemed to be less harmful than previously reported with careful adaptation, control and follow-ups.

Increased microvascular permeability in mice lacking Epac1 (Rapgef3).

AIM: Maintenance of the blood and extracellular volume requires tight control of endothelial macromolecule permeability, which is regulated by cAMP signalling. This study probes the role of the cAMP mediators rap guanine nucleotide exchange factor 3 and 4 (Epac1 and Epac2) for in vivo control of microvascular macromolecule permeability under basal conditions. METHODS: Epac1-/- and Epac2-/- C57BL/6J mice were produced and compared with wild-type mice for transvascular flux of radio-labelled albumin in skin, adipose tissue, intestine, heart and skeletal muscle. The transvascular leakage was also studied by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) using the MRI contrast agent Gadomer-17 as probe. RESULTS: Epac1-/- mice had constitutively increased transvascular macromolecule transport, indicating Epac1-dependent restriction of baseline permeability. In addition, Epac1-/- mice showed little or no enhancement of vascular permeability in response to atrial natriuretic peptide (ANP), whether probed with labelled albumin or Gadomer-17. Epac2-/- and wild-type mice had similar basal and ANP-stimulated clearances. Ultrastructure analysis revealed that Epac1-/- microvascular interendothelial junctions had constitutively less junctional complex. CONCLUSION: Epac1 exerts a tonic inhibition of in vivo basal microvascular permeability. The loss of this tonic action increases baseline permeability, presumably by reducing the interendothelial permeability resistance. Part of the action of ANP to increase permeability in wild-type microvessels may involve inhibition of the basal Epac1-dependent activity.

Long-term consumption of an obesogenic high fat diet prior to ischemia-reperfusion mediates cardioprotection via Epac1-dependent signaling.

BACKGROUND: Obesity is still considered a risk factor for cardiovascular disease, although more recent knowledge also suggests obesity to be associated with reduced morbidity and mortality - the "obesity paradox". This study explores if long-term feeding of an obesogenic high fat diet renders the myocardium less susceptible to ischemic-reperfusion induced injury via Epac-dependent signaling. METHODS: Wild type (wt), Epac1 (Epac1-/-) and Epac2 (Epac2-/-) deficient mice were fed a high fat (HFD) or normal chow diet (ND) for 33 ± 1 weeks. Six experimental groups were included: (1) control wt ND (wt ND), (2) control wt HFD (wt HFD), (3) Epac1-/- mice on ND (Epac1-/-ND), (4) Epac1-/- mice on HFD (Epac1-/-HFD), (5) Epac2-/- mice on ND (Epac2-/-ND), and (6) Epac2-/- mice on HFD (Epac2-/-HFD). Isolated ex vivo mice hearts were perfused in a constant pressure Langendorff mode, and exposed to 30min of global ischemia (GI) and 60min of reperfusion. Endpoints were infarct size and functional recovery. RESULTS: All groups fed a HFD presented with significantly enhanced body weight, visceral fat content and reduced glucose clearance compared to corresponding ND groups. Although the HFD cohorts presented with an overall comparable systemic capability to clear glucose, the Epac1-/- HFD group presented with glucose levels slightly above the human diabetes criteria at the end of the intraperitoneal glucose tolerance test (ipGTT). Moreover, the HFD significantly reduced infarct size in both wild type (wt HFD 41.3 ± 5.5% vs. wt ND 58.0 ± 9.8%, p < 0.05) and Epac2-/- cohorts (Epac2-/-HFD 34.4 ± 7.2% vs. Epac2-/-ND 56.5 ± 3.8%, p < 0.05). Interestingly, however, the HFD did not reduce infarct size in Epac1-/- deficient mice hearts (Epac1-/-HFD 65.1 ± 5.1% vs. Epac1-/-ND 56.1 ± 3.5%, ns.). CONCLUSION: Epac1-dependent signaling is involved in mediating the cardioprotection afforded by long-term feeding of an obesogenic high fat diet in mice hearts.

Repeated treatments of drooling with botulinum toxin B in neurology.

OBJECTIVES: To investigate efficacy, saliva flow, and composition in repeated BoNT-B treatments of drooling. MATERIALS AND METHODS: Seventeen neurological patients (median 66 years), referred for treatment of drooling participated in this observational study. Median total doses of 4000 units botulinum toxin type B (BoNT-B, Neurobloc(®)) were injected with at least 3 months intervals into parotid and submandibular glands using ultrasound guidance. Measures of drooling and saliva collection for analysis were obtained before treatment, and 6, 12, and eventually 18 weeks after. RESULTS: Number of treatment series in each patient was 1-7. Compared to baseline, saliva flow rate and drooling were reduced 30-70% 6 weeks after treatment in the first series, while sodium, chloride, and total protein increased 20-80% (t-tests; P < 0.05). After 12 weeks, drooling was still significantly reduced, saliva flow tended to be, and saliva composition was back to baseline. Frequent side effects were viscous saliva and dry mouth. Due to fading effect in eight patients, individual decisions were taken to change from BoNT-B to BoNT-A. Similarly, the outcome was significantly reduced over time in six patients completing five subsequent BoNT-B treatment series (ANOVA; P < 0.05). CONCLUSION: In the first series, BoNT-B treatment resulted in marked reduction of drooling and saliva flow rate with some relapse after 12 weeks. The viscous saliva was ascribed to increased total protein content and compensatory mechanisms related to ß-adrenergic receptor-specific actions. With patients needing long-term treatment, it should be noted that the efficacy of repeated BoNT-B may fade with time.

Jaw muscle soreness after tooth-clenching depends on force level.

The effects of prolonged tooth-clenching on masticatory muscle pain have not been fully elucidated. We hypothesized that late-onset soreness may develop depending on the clenching force. Ten pain-free females were asked to endure sustained clenching tasks up to exhaustion in randomized sequences of 7.5%, 10%, 15%, 25%, and 40% of maximum clenching force. Perceived pain, fatigue, and pressure-pain thresholds of masseter and temporalis muscles were assessed before, immediately after, and one day after the tasks. Endurance times differed markedly among participants and force levels, ranging from 1.2 to 245.1 min. Masseter pressure-pain threshold decreased immediately after (-13.7%; p = 0.050) and one day after (-22.0%; p = 0.006) the 7.5% task. Temporalis pressure threshold decreased one day after the 7.5 % task (-14.6%; p = 0.003). It was concluded that prolonged low-level tooth-clenching in healthy young women induces a delayed soreness in the jaw elevator muscles.

Reduced jaw opening from paradoxical activity of mandibular elevator muscles treated with botulinum toxin.

The aim of the study was the effect of injections with botulinum toxin A (BTX-A) on reduced jaw opening, caused by paradoxical, antagonistic activity of jaw elevator muscles after brain stem lesions. The study included a male (51 years) and a female (69 years) patient. Subjective assessment, clinical recordings, muscle blocks and electromyography (EMG) were used to diagnose paradoxical activity, and to plan, guide and evaluate the treatment. The paradoxical innervation pattern was unilateral in the male and bilateral in the female. The paradoxical activity during jaw opening amounted to 24-109% of the level during maximum biting, and bursts of paradoxical activity were also present during chewing. EMG-guided blocks and later BTX-A injections of the affected muscles increased the opening by 9-23 mm from pre-treatment values of 15-18 mm, and normalized chewing. The study proved BTX-A to be an effective treatment for reduced jaw opening caused by paradoxical activity. Treatment was optimized by EMG evaluation of the current activity of the jaw elevator muscles, permitting individual treatment plans with longer intervals between BTX-A injections and lower doses than with conventional treatment for oromandibular dystonia. Thus the treatment only had to be repeated one to two times per year to maintain acceptable jaw mobility.

A three-dimensional method of visualizing the temporomandibular joint based on magnetic resonance imaging in a case of juvenile chronic arthritis.

AIM: The purpose of this study was to present a new visualizing method for temporomandibular joint (TMJ) pathology in 3D at several condylar positions and to apply the method to a case of juvenile chronic arthritis (JCA) with previously diagnosed bilateral TMJ disorder and to a subject without signs and symptoms from the TMJ. SUBJECT: The 20-year-old female patient had suffered from polyarticular JCA from the age of 6 years 8 months. The present study is based on a follow-up examination after the completion of orthodontic treatment with the Herbst appliance. Both TMJs were examined using magnetic resonance imaging (MRI) at closed and open mouth positions. The mandibular condyle, the glenoid fossa and the articular disc were segmented and 3D reconstruction of these structures was carried out. RESULTS: The condyle was characterized by an increased sagittal diameter with osteophyte-like formation at the anterior aspect of the condylar head and flattening of the superior surface. The depth of the glenoid fossa was reduced and the articular eminence was remarkably flat. The articular disc was markedly diminished and posteriorly placed on the condylar head. The distance of the condylar path during mouth opening was reduced and the curvature of the condylar path was quite flat. The disc moved slightly posteriorly on the condylar head and remained in the glenoid fossa during mouth opening. CONCLUSION: The imaging modality described improves visualization of TMJ morphology and gains insight into the TMJ pathology of the JCA patient, adding to understanding of the clinical problems.

Pituitary-specific knockout of steroidogenic factor 1.

Knockout mice lacking the orphan nuclear receptor steroidogenic factor 1 (SF-1) revealed its essential roles at multiple levels of endocrine development and function. These SF-1 knockout mice lacked adrenal glands and gonads, thereby manifesting adrenal insufficiency and sex reversal of their internal and external genitalia. Their pituitary gonadotropes failed to express several markers of normal differentiated function, and they lacked a specific hypothalamic nucleus, the ventromedial hypothalamic nucleus (VMH). Using the Cre-loxP system, we generated mice whose gene encoding SF-1 was inactivated specifically in the anterior pituitary. These pituitary-specific SF-1 knockout mice were sterile and never matured sexually. Their gonads weighed only approximately 5% of the weight of wild-type gonads. SF-1 immunoreactivity was absent in the anterior pituitary but was unaffected in the adrenal cortex, validating the selectivity of the gene targeting strategy. Consistent with an important role of SF-1 in gonadotropes, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were markedly decreased in the pituitary-specific SF-1 knockout mice. The pituitary-specific SF-1 knockout mice are a novel genetic model of hypogonadotropic hypogonadism and establish essential roles of SF-1 in gonadotropin expression.

Orofacial pain, jaw function, and temporomandibular disorders in women with a history of juvenile chronic arthritis or persistent juvenile chronic arthritis.

OBJECTIVES: We sought to study the long-term outcome of juvenile chronic arthritis (JCA) in the temporomandibular joint (TMJ). STUDY DESIGN: Temporomandibular disorders, including TMJ involvement, were assessed in 42 women with pauciarticular or polyarticular JCA--on average 25.8 years from disease onset--and compared with those found in matched control subjects. Disease-related parameters associated with temporomandibular disorders were identified. RESULTS: The TMJ was involved in 66.7% of the patients, most severely in extended pauciarticular JCA. Temporomandibular disorders were more frequent in the patients than in the control subjects, especially in those with persistent disease. The TMJ involvement was positively correlated with disease duration and negatively correlated with jaw opening and occlusal support. Duration of active JCA and history of functional pain were identified as predictors of present TMJ involvement. CONCLUSION: In a long-term follow-up, TMJ involvement proved frequent in the studied patients and was associated with long disease duration and previous pain on jaw opening. The findings suggest that patients with JCA should undergo orofacial evaluation on a regular basis.

Temporomandibular disorders in relation to craniofacial dimensions, head posture and bite force in children selected for orthodontic treatment.

The present study examined the associations between craniofacial dimensions, head posture, bite force, and symptoms and signs of temporomandibular disorders (TMD). The sample comprised 96 children (51F, 45M) aged 7-13 years, sequentially admitted for orthodontic treatment of malocclusions entailing health risks. Symptoms and signs of TMD were assessed by 37 variables describing the occurrence of headache and facial pain, clicking, jaw mobility, tenderness of muscles and joints, and the Helkimo Anamnestic and Dysfunction indices. Craniofacial dimensions (33 variables), and head and cervical posture (nine variables) were recorded from lateral cephalometric radiographs taken with the subject standing with the head in a standardized posture (mirror position). Dental arch widths were measured on plaster casts and bite force was measured at the first molars on each side by means of a pressure transducer. Associations were assessed by Spearman correlations and multiple stepwise logistic regression analyses. The magnitudes of the significant associations were generally low to moderate. On average, temporomandibular joint (TMJ) dysfunction was seen in connection with a marked forward inclination of the upper cervical spine and an increased craniocervical angulation, but no firm conclusion could be made regarding any particular craniofacial morphology in children with symptoms and signs of TMJ dysfunction. Muscle tenderness was associated with a 'long face' type of craniofacial morphology and a lower bite force. Headache was associated with a larger maxillary length and increased maxillary prognathism. A high score on Helkimo's Clinical Dysfunction Index was associated with smaller values of a number of vertical, horizontal, and transversal linear craniofacial dimensions and a lower bite force.

Hypomorphic phenotype in mice with pituitary-specific knockout of steroidogenic factor 1.

The bacteriophage Cre recombinase provides a powerful approach for tissue-specific gene inactivation. Using a Cre transgene driven by the common alpha subunit of glycoprotein hormones (alphaGSU-Cre), we have previously inactivated steroidogenic factor 1 (SF-1) in the anterior pituitary, causing hypogonadotropic hypogonadism with sexual infantilism, sterility, and severe gonadal hypoplasia. We now explore the molecular mechanisms underlying a hypomorphic gonadal phenotype in mice carrying two floxed SF-1 alleles (F/F) relative to mice carrying one recombined and one floxed allele (F/R). Because their Cre-mediated disruption of the locus encoding SF-1 was less efficient, alphaGSU-Cre, F/F mice retained some gonadotropin-expressing cells in the anterior pituitary, thereby stimulating some gonadal function. This novel in vivo model for exploring the effects of differing levels of gonadotropins on gonadal development highlights the need for careful genotype-phenotype comparisons in studies using Cre recombinase to produce tissue-specific knockouts.

Approaches to define the role of SF-1 at different levels of the hypothalamic-pituitary-steroidogenic organ axis.

Targeted gene disruption has produced knockout mice lacking the orphan nuclear receptor steroidogenic factor 1 (SF-1). These SF-1 knockout mice lacked adrenal glands and gonads, resulting in adrenocortical insufficiency and sex reversal of their internal and external genitalia. They also had impaired expression of pituitary gonadotropins and agenesis of the ventromedial hypothalamic nucleus (VMH), confirming roles of SF-1 at multiple levels of the hypothalamic-pituitary-steroidogenic tissue axis. Using the Cre-loxP system, we now have generated mice in which SF-1 is inactivated selectively in the anterior pituitary. These pituitary-specific SF-1 knockout mice were sterile and failed to exhibit sexual maturation. Histologically, their gonads were markedly hypoplastic, weighing only approximately 5% of the gonads of wild-type mice. Consistent with an important role of SF-1 in gonadotropes, there were no cells in the pituitary gland that expressed either follicle-stimulating hormone (FSH) or luteinizing hormone (LH). These pituitary-specific SF-1 knockout mice are a novel genetic model of hypogonadotropic hypogonadism and establish essential roles of SF-1 in gonadotropin expression.

Effects of prolonged gum chewing on pain and fatigue in human jaw muscles.

Gum chewing has been accepted as an adjunct to oral hygiene, as salivary stimulant and vehicle for various agents, as well as for jaw muscle training. The aim of this study was to investigate the effects of prolonged gum chewing on pain, fatigue and pressure tenderness of the masticatory muscles. Fifteen women without temporomandibular disorders (TMD) were requested to perform one of the following chewing tasks in three separate sessions: chewing a very hard gum, chewing a soft gum, and empty-chewing with no bolus. Unilateral chewing of gum or empty chewing was performed for 40 min at a constant rate of 80 cycles/min. In each session, perceived muscle pain and masticatory fatigue were rated on visual analog scales (VAS) before, throughout, and after the chewing task. Pressure pain thresholds (PPTs) of masseter and anterior temporalis muscles were assessed before and immediately after the chewing tasks, and again after 24 h. The VAS scores for pain and fatigue significantly increased only during the hard gum chewing, and after 10 min of recovery VAS scores had decreased again, almost to their baseline values. No significant changes were found for PPTs either after hard or soft gum chewing. The findings indicate that the jaw muscles recover quickly from prolonged chewing activity in subjects without TMD.

Effects of ethnicity, gender and age on clenching force and load distribution.

The aim of the study was to investigate the effect of ethnicity, gender and age on the degree of clenching force and distribution of the load over the dentition. The maximal clenching force was measured in 12 young Danish females, 12 young Japanese females, 12 young Japanese males and 10 senior Japanese males using the Dental Prescale System. The occlusal contact area, average pressure, occlusal load, antero-posterior and right-left location of the occlusal load center (i.e., a center of balance of the occlusal load distributed over the maxillary dentition: OLC) and dental arch length and width were analyzed and compared. The arch width and average pressure in young Danish females were significantly smaller/lower than in Japanese females. The arch length, arch width, contact area and total occlusal load in young Japanese males were significantly greater than those of the Japanese females. The average pressure in young Japanese males was also greater than in senior males. The location of the OLC was almost the same in all groups. The results suggest that the analysis of the OLC may be a useful method for evaluating occlusal function and prosthodontic treatment because the location of the OLC is not affected by ethnicity, gender and age.

SF-1: a critical mediator of steroidogenesis.

Studies in knockout mice have established that the orphan nuclear receptor steroidogenic factor 1 (SF-1) plays essential roles in the development and function of the primary steroidogenic organs. These SF-1 knockout mice lacked adrenal glands and gonads, causing adrenocortical insufficiency and sex reversal of their internal and external genitalia. They also had impaired expression of pituitary gonadotropins and agenesis of the ventromedial hypothalamic nucleus (VMH), confirming roles of SF-1 at all three levels of the hypothalamic-pituitary-steroidogenic organ axis. Ongoing experiments are directed at developing methods to inactivate SF-1 in a tissue-specific manner.

Bite force in pre-orthodontic children with unilateral crossbite.

In the present study bite force was examined in pre-orthodontic children with unilateral posterior crossbite and compared with an age- and sex-matched control group. The sample comprised 52 children aged 7-13 years, 26 pre-orthodontic children with unilateral posterior crossbite (crossbite group), and 26 children with neutral occlusion (control group). Unilateral bite force was measured at the first molar by means of a pressure transducer. Furthermore, symptoms and signs of temporomandibular disorders (TMD) and number of teeth in contact in the intercuspal position (ICP) were recorded. In both groups, the maximum bite force increased significantly with age and with increasing stages of dental eruption, but the bite force in both sexes did not differ significantly. There were no significant differences in bite force between sides, but this was significantly smaller in the crossbite group than in the controls (P < 0.001). Regression analysis showed that stage of dental eruption (P < 0.001), number of teeth in occlusal contact (P < 0.01), and unilateral crossbite (P < 0.001) were the only variables significantly correlated with bite force. The number of teeth in contact was significantly smaller in the crossbite group than in the controls (P < 0.05) and the frequency of muscle tenderness was significantly higher in the crossbite group than in the controls (P < 0.05). These results suggest that differences in the muscle function associated with unilateral crossbite lead to a significantly smaller bite force in the crossbite group compared with controls and this difference did not diminish with age and development. These findings indicate that early treatment of unilateral posterior crossbite is advisable to optimize conditions for function.

Steroidogenic factor 1 (SF1) is essential for pituitary gonadotrope function.

Knockout mice lacking the orphan nuclear receptor steroidogenic factor 1 (SF1) exhibit a complex endocrine phenotype that includes adrenal and gonadal agenesis, impaired expression of pituitary gonadotropins, and absence of the ventromedial hypothalamic nucleus (VMH). These multiple defects complicate efforts to delineate primary versus secondary effects of SF1 deficiency in different tissues, such that its direct role in gonadotropes remains uncertain. To define this role, we have expressed Cre recombinase driven by the promoter region of the common alpha subunit of glycoprotein hormones (alpha GSU), thereby inactivating a loxP-modified SF1 locus in the anterior pituitary gland. Although pituitary-specific SF1 knockout mice were fully viable, they were sterile and failed to develop normal secondary sexual characteristics. Their adrenal glands and VMH appeared normal histologically, but their testes and ovaries were severely hypoplastic. alpha GSU-Cre, loxP mice had normal levels of most pituitary hormones, but had markedly decreased expression of LH and FSH. Treatment with exogenous gonadotropins stimulated gonadal steroidogenesis, inducing germ cell maturation in males and follicular and uterine maturation in females--establishing that the gonads can respond to gonadotropins. The pituitary-specific SF1 knockout mice are a novel genetic model of hypogonadotropic hypogonadism that establishes essential role(s) of SF1 in pituitary gonadotropes.

Quantitative ultrasound image analysis of the supraspinatus muscle.

OBJECTIVE: To develop and validate a method, based on quantitative ultrasound image analysis, to objectively analyse and characterize the ultrasound images of m. supraspinatus. DESIGN: Quantitative ultrasonography was performed on the supraspinatus muscle of 14 healthy subjects. METHODS: A computerized analysis using first-order grey-scale statistics to evaluate the muscle tissue composition was developed and validated. RESULTS: Data from one scanning site were not representative for the whole muscle due to muscle inhomogenity. Using first-order grey-scale statistics the scanning direction was of no importance. By using a scanning session consisting of three different scanning sites along the muscle in two directions, longitudinally and transversely, to characterize the tissue composition of the muscle, a high day-to-day reproducibility was obtained. CONCLUSION: The described scanning session is a relatively sensitive and reproducible method for studying the muscle tissue composition. RelevanceQuantitative ultrasonography seems to be a potential clinical and occupational examination method to detect tissue composition of myalgic muscles compared to healthy muscles.