PUM1 and PGK1 are Favorable Housekeeping Genes over Established Biodosimetry-related Housekeeping Genes such as HPRT1, ITFG1, DPM1, MRPS5, 18S rRNA and Others after Radiation Exposure.

In gene expression (GE) studies, housekeeping genes (HKGs) are required for normalization purposes. In large-scale inter-laboratory comparison studies, significant differences in dose estimates are reported and divergent HKGs are employed by the teams. Among them, the 18S rRNA HKG is known for its robustness. However, the high abundance of 18S rRNA copy numbers requires dilution, which is time-consuming and a possible source of errors. This study was conducted to identify the most promising HKGs showing the least radiation-induced GE variance after radiation exposure. In the screening stage of this study, 35 HKGs were analyzed. This included selected HKGs (ITFG1, MRPS5, and DPM1) used in large-scale biodosimetry studies which were not covered on an additionally employed pre-designed 96-well platform comprising another 32 HKGs used for different exposures. Altogether 41 samples were examined, including 27 ex vivo X-ray irradiated blood samples (0, 0.5, 4 Gy), six X-irradiated samples (0, 0.5, 5 Gy) from two cell lines (U118, A549), as well as eight non-irradiated tissue samples to encompass multiple biological entities. In the independent validation stage, the most suitable candidate genes were examined from another 257 blood samples, taking advantage of already stored material originating from three studies. These comprise 100 blood samples from ex vivo X-ray irradiated (0-4 Gy) healthy donors, 68 blood samples from 5.8 Gy irradiated (cobalt-60) Rhesus macaques (RM) (LD29/60) collected 0-60 days postirradiation, and 89 blood samples from chemotherapy-(CTx) treated breast tumor patients. CTx and radiation-induced GE changes in previous studies appeared comparable. RNA was isolated, converted into cDNA, and GE was quantified employing TaqMan assays and quantitative RT-PCR. We calculated the standard deviation (SD) and the interquartile range (IQR) as measures of GE variance using raw cycle threshold (Ct) values and ranked the HKGs accordingly. Dose, time, age, and sex-dependent GE changes were examined employing the parametrical t-test and non-parametrical Kruskal Wallis test, as well as linear regression analysis. Generally, similar ranking results evolved using either SD or IQR GE measures of variance, indicating a tight distribution of GE values. PUM1 and PGK1 showed the lowest variance among the first ten most suitable genes in the screening phase. MRPL19 revealed low variance among the first ten most suitable genes in the screening phase only for blood and cells, but certain comparisons indicated a weak association of MRPL19 with dose (P = 0.02-0.09). In the validation phase, these results could be confirmed. Here, IQR Ct values from, e.g., X-irradiated blood samples were 0.6 raw Ct values for PUM1 and PGK1, which is considered to represent GE differences as expected due to methodological variance. Overall, when compared, the GE variance of both genes was either comparable or lower compared to 18S rRNA. Compared with the IQR GE values of PUM1 and PGKI, twofold-fivefold increased values were calculated for the biodosimetry HKG HPRT1, and comparable values were calculated for biodosimetry HKGs ITFG1, MRPS5, and DPM1. Significant dose-dependent associations were found for ITFG1 and MRPS5 (P = 0.001-0.07) and widely absent or weak (P = 0.02-0.07) for HPRT1 and DPM1. In summary, PUM1 and PGK1 appeared most promising for radiation exposure studies among the 35 HKGs examined, considering GE variance and adverse associations of GE with dose.

Development of a Point-of-Care Microfluidic RNA Extraction Slide for Gene Expression Diagnosis after Irradiation.

In times of war, radiological/nuclear emergency scenarios have become a reemphasized threat. However, there are challenges in transferring whole-blood samples to laboratories for specialized diagnostics using RNA. This project aims to miniaturize the process of unwieldy conventional RNA extraction with its stationed technical equipment using a microfluidic-based slide (MBS) for point-of-care diagnostics. The MBS is thought to be a preliminary step toward the development of a so-called lab-on-a-chip microfluidic device. A MBS would enable early and fast field care combined with gene expression (GE) analysis for the prediction of hematologic acute radiation syndrome (HARS) severity or identification of RNA microbes. Whole blood samples from ten healthy donors were irradiated with 0, 0.5 and 4 Gy, simulating different ARS severity degrees. RNA quality and quantity of a preliminary MBS was compared with a conventional column-based (CB) RNA extraction method. GE of four HARS severity-predicting radiation-induced genes (FDXR, DDB2, POU2AF1 and WNT3) was examined employing qRT-PCR. Compared to the CB method, twice as much total RNA from whole blood could be extracted using the MBS (6.6 ± 3.2 µg vs. 12.0 ± 5.8 µg) in half of the extraction time, and all MBS RNA extracts appeared DNA-free in contrast to the CB method (30% were contaminated with DNA). Using MBS, RNA quality [RNA integrity number equivalent (RINe)] values decreased about threefold (3.3 ± 0.8 vs. 9.0 ± 0.4), indicating severe RNA degradation, while expected high-quality RINe ≥ 8 were found using column-based method. However, normalized cycle threshold (Ct) values, as well as radiation-induced GE fold-changes appeared comparable for all genes utilizing both methods, indicating that no RNA degradation took place. In summary, the preliminary MBS showed promising features such as: 1. halving the RNA extraction time without the burden of heavy technical equipment (e.g., a centrifuge); 2. absence of DNA contamination in contrast to CB RNA extraction; 3. reduction in blood required, because of twice the biological output of RNA; and 4. equal GE performance compared to CB, thus, increasing its appeal for later semi-automatic parallel field applications.

THE RESULTS OF THE EURADOS INTERCOMPARISON IC2014 FOR WHOLE-BODY DOSEMETERS IN PHOTON FIELDS.

The European Dosimetry Group (EURADOS) first started performing international intercomparisons for whole-body dosemeters for individual monitoring services in 1998. Since 2008, these whole-body intercomparisons have been performed on a regular basis. In this latest intercomparison (IC2014), 96 monitoring services from 35 countries (mostly European) participated with 112 dosimetry systems. Unlike in the previous intercomparisons, the whole registration, communication and data exchange process was handled by a new on-line platform. All dosemeter irradiations were carried out in the Seibersdorf accredited dosimetry laboratory. The irradiation plan consisted of nine irradiation setups with five different photon radiation qualities (S-Cs, S-Co, RQR7, W-80 and W-150) and two different angles of radiation incidence (0° and 60°). The paper describes and analyses the individual results for the personal dose equivalent quantities Hp(10) and if requested, Hp(0.07), for all participating systems and compares these results with the ISO 14146 'trumpet curve' performance criteria. The results show that 100 systems (89 % of all systems) do fulfil the general ISO 14146 performance criteria. This paper gives an overview on the performance of the participating individual monitoring services and the influence of the dosemeter type on the observed response values.

[Imitation of an Anderson type II dens fracture by a motion artefact in computed tomography : Four case examples]. / Imitation einer Densfraktur Typ Anderson II durch ein computertomographisches Bewegungsartefakt : Vier Fallbeispiele.

Computed tomography (CT) is the method of choice in the diagnosis and classification of odontoid fractures with a sensitivity of more than 99 % and a specificity almost equally as high. In this article we report on four cases where CT-generated motion artefacts exactly mimicked an Anderson type II fracture of the dens axis, initially leading to a wrong diagnosis. Although this seems to be a very rare event, these cases indicate that overlooked CT motion artefacts can lead to severe consequences and attention must be paid to the radiological signs outlined in this report.

Survey of dental clinic patients: smoking and preferences for cessation support.

BACKGROUND: Smoking cessation interventions delivered by dental practitioners can be as effective as those delivered by general medical practitioners. However, concern that addressing smoking may cause offence to their patients is a reason cited by dental practitioners for not regularly addressing patient smoking behaviours, despite believing they should play a role in smoking cessation. This study aimed to elicit the smoking behaviour and smoking cessation preferences of dental patients to determine if these concerns accurately reflect patient attitudes. METHODS: We surveyed 726 adult dental patients attending The University of Queensland's School of Dentistry dental clinics, Brisbane Dental Hospital and four private dental practices in South-East Queensland. RESULTS: Most (80%) current daily smokers had tried to quit smoking. Smokers and non-smokers both agreed that dentists should screen for smoking behaviour and are qualified to offer smoking cessation advice (99% and 96% respectively). Almost all participants (96%) said they would be comfortable with their dentist asking about their smoking and that if their smoking was affecting their oral health their dentist should advise them to quit. CONCLUSIONS: Patients are receptive to dental practitioners inquiring about smoking behaviour and offering advice on quitting. Smoking patients showed considerable motivation and interest in quitting smoking, particularly in the context of health problems related to smoking being identified. These results should encourage dentists to raise the issue with their patients.

Sex specific impact of prodromal chest pain on pre-hospital delay time during an acute myocardial infarction: Findings from the multicenter MEDEA Study with 619 STEMI patients.

BACKGROUND: Scarce evidence yields conflicting results regarding the effect of prodromal chest pain (PCP) on pre-hospital delay during an acute myocardial infarction (AMI). We aimed to assess the impact of PCP on delay. METHODS: Data was collected on 619 ST-elevated MI patients from the multicenter Munich Examination of Delay in Patients Experiencing Acute Myocardial Infarction (MEDEA) study. Patients with any PCP (which was subdivided into undefined PCP, possible and definite angina) within a year before AMI were identified using the Rose questionnaire, administered in bedside interviews. The influence of PCP and its subdivisions (all compared to no PCP) was assessed using logistic regression (with cut-offs of 2 h, 6 h, and a 4-category ordinal outcome). RESULTS: Any type of PCP was reported by men (50.6%) more than women (34.6%) (OR=1.9; 95% CI: 1.3 to 2.8; p=.001). The median delay of patients with PCP was not significantly different to delay in patients with no PCP (p=.327). Prolonged delay times were observed in women with PCPs of lesser degree of cardiac confirmation, while the opposite was observed in men. In women, possible angina was more strongly associated with delay <2 h (OR=6.8; 95% CI=2 to 23.8) than any PCP (OR=2.6; 95% CI=1.2 to 5.7). CONCLUSIONS: For men, PCPs of increasing cardiac confirmation are associated with prolonged delay. For women, PCPs of lesser cardiac confirmation are more likely to lead to prolonged delay. Future studies should investigate mediating factors.

Survey of Australian oral health practitioners and their smoking cessation practices.

BACKGROUND: Smoking is a significant health and economic burden in Australia. Studies of smoking cessation practices in dental settings have primarily concentrated on dentists rather than other oral health practitioner (OHP) groups (dental hygienists, dental therapists and oral health therapists). The aim of this study was to measure Australian OHPs' attitudes, behaviours, interest and barriers to delivering smoking cessation interventions. METHODS: Members of the two peak professional bodies representing Australian OHPs were invited to participate in an anonymous online questionnaire. RESULTS: There were discrepancies between practitioner attitudes and current smoking cessation practices. While the majority of practitioners (90.1%) frequently screened for smoking behaviour, fewer (51.1%) assisted patients to quit smoking. The principal form of assistance was referral to Quitline (45.7%) or to a general medical practitioner (44.4%). The most prevalent barriers identified were lack of knowledge of pharmacological treatments (45.8%) and lack of access to smoking cessation resources (44.2%). Contrary to international studies, time and financial incentive were not commonly cited barriers to delivering smoking cessation interventions. CONCLUSIONS: This survey identifies a need for continuing education in smoking cessation practice. Dissemination of policies, guidelines and resources may assist OHPs to become more engaged and confident in delivering smoking cessation interventions as part of their routine practice.

[Chronic knee pain in children and adolescents: review of anatomical and overload-related knee pain]. / Chronische Knieschmerzen bei Kindern und Jugendlichen : Eine Übersicht über anlage- und überlastungsbedingte Knieschmerzen.

BACKGROUND: Knee pain in children and adolescents is a common reason for presentation in pediatric orthopedic consultation. The causes are manifold and require a thorough patient history, detailed physical examination and extensive diagnostics. DIAGNOSTICS: Chronic knee pain in children and adolescents is a diagnosis by exclusion. An extensive patient history including training habits, pain localization and clinical examination provide indications vital for the diagnosis. Overuse is often the trigger. In dynamic investigations consideration should be given to muscle imbalance. Imaging techniques have a high sensitivity and specificity only in combination with the anamnesis and clinical findings. THERAPY: With pain therapy, targeted physiotherapy, a break in active sport or training modifications, the prognosis is good for overload syndromes. An operative therapy after unsuccessful conservative therapy is necessary in only a few cases. Among the anatomy-related types of knee pain primary popliteal cysts mostly require no therapy and for plica syndrome arthroscopic resection should only be necessary if conservative treatment is unsuccessful. Operative therapy is necessary for symptomatic discoid meniscus.

A longitudinal study of the bi-directional relationship between tobacco smoking and psychological distress in a community sample of young Australian women.

BACKGROUND: Tobacco smoking and poor mental health are both prevalent and detrimental health problems in young women. The temporal relationship between the two variables is unclear. We investigated the prospective bi-directional relationship between smoking and mental health over 13 years. METHOD: Participants were a randomly selected community sample of 10 012 young women with no experience of pregnancy, aged 18-23 years at baseline (1996) from the Australian Longitudinal Study on Women's Health. Follow-up surveys over 13 years were completed in 2000, 2003, 2006 and 2009, allowing for five waves of data. Measures included self-reported smoking and mental health measured by the Mental Health Index from the 36-item short-form health questionnaire and the 10-item Center for Epidemiologic Studies Depression Scale. Sociodemographic control variables included marital status, education level and employment status. RESULTS: A strong cross-sectional dose-response relationship between smoking and poor mental health was found at each wave [odds ratio (OR) 1.41, 95% confidence intervals (CI) 1.17-1.70 to OR 2.27, 95% CI 1.82-2.81]. Longitudinal results showed that women who smoked had 1.21 (95% CI 1.06-1.39) to 1.62 (95% CI 1.24-2.11) times higher odds of having poor mental health at subsequent waves. Women with poor mental health had 1.12 (95% CI 1.17-1.20) to 2.11 (95% CI 1.68-2.65) times higher odds of smoking at subsequent waves. These results held after adjusting for mental health history and smoking history and sociodemographic factors. Correlation analysis and structural equation modelling results were consistent in showing that both directions of the relationship were statistically significant. CONCLUSIONS: The association between poor mental health and smoking in young women appeared to be bi-directional.

Are Australian smokers interested in using low-nitrosamine smokeless tobacco for harm reduction?

AIMS: To determine (1) whether Australian smokers are aware of low-nitrosamine smokeless tobacco (LNSLT) products and (2) whether they would be interested in using LNSLT either as a long-term substitute for smoking or as an aid to quitting, if these products were to become legally available. METHODS: 401 daily smokers were recruited by a market research company to complete an internet questionnaire about their smoking history, knowledge of smokeless tobacco and intentions to purchase LNSLT under different scenarios. FINDINGS: Just under half (48%) indicated they were willing to buy an LNSLT product. Predictors of an interest in purchasing LNSLT were low income, poorer health, prior SLT use, belief that SLT is less harmful than cigarettes, switching to a lower tar cigarette in the past year, ever using nicotine replacement therapy products for quitting or other reasons, having made a failed cessation attempt in the previous year and not planning to quit smoking. Analysis of quitting and LNSLT purchasing intentions under different scenarios suggest that making LNSLT available at a much lower cost than smoked cigarettes while increasing taxes on cigarettes could provide a greater reduction in the number of smokers than the same tax increase alone. These results support further examination of the potential for LNSLT to reduce smoking-related harm in Australia.

Supping with the devil? The role of law in promoting tobacco harm reduction using low nitrosamine smokeless tobacco products.

In Sweden, male cigarette smoking has declined as snus, a smokeless tobacco product which is low in carcinogenic nitrosamines, has gained popularity among male tobacco users. Epidemiological modelling based on the Swedish experience indicates that there would be major public health gains if a substantial number of current smokers in other countries could also be persuaded to switch to this product. This form of 'tobacco harm reduction' is very controversial in the public health community for many reasons. These include: objections in principle to the use of less harmful but still addictive nicotine products; uncertainties about the long-term effects of these products on health; doubts about the likely interest in and uptake of these products among existing smokers; concerns that increasing the availability of these products will increase the number of new tobacco users and eventually the number of smokers in the population; and anxiety about how the tobacco industry may use these products to undermine current tobacco control policies. This paper concludes with suggestions for a graduated series of policies that may allow exploration of the public health costs and benefits of encouraging smokers to switch to snus.

Predicting the future prevalence of cigarette smoking in Australia: how low can we go and by when?

BACKGROUND: In Australia, smoking prevalence has declined in men since the 1950s and in women since the 1980s. Future smoking prevalence in Australia is predicted from estimates of previous and current age-specific and sex-specific cessation rates and smoking uptake in young people derived from national survey data on the prevalence of smoking between 1980 and 2007. METHODS: A dynamic forecasting model was used to estimate future smoking prevalence in the Australian population based on a continuation of these current trends in smoking uptake and cessation. RESULTS: The results suggest that Australia's smoking prevalence will continue to fall while current rates of initiation and cessation are maintained. But a continuation of current smoking cessation and initiation patterns will see around 14% of adults still smoking in 2020. CONCLUSIONS: Smoking cessation rates will need to double for Australian smoking prevalence to reach a policy target of 10% by 2020.

Inhibition of p38 mitogen-activated protein kinase attenuates left ventricular dysfunction by mediating pro-inflammatory cardiac cytokine levels in a mouse model of diabetes mellitus.

AIMS/HYPOTHESIS: We investigated the effect of SB 203580, a pharmacological inhibitor of p38 mitogen-activated protein kinase (MAPK), on cardiac inflammation, cardiac fibrosis, and left ventricular function using an animal model of diabetic cardiomyopathy. MATERIALS AND METHODS: Diabetes mellitus was induced by streptozotocin (50 mg/kg i.p. for 5 days) in 20 C57/BL6J mice. Diabetic mice were treated daily with the p38 MAPK inhibitor SB 203580 (1 mg/kg daily, n=10) or with placebo (n=10) and were compared to non-diabetic controls. Left ventricular function was measured by pressure-volume loops after 8 weeks of diabetes mellitus. The parameters for systolic function were the end systolic pressure-volume relationship (ESPVR) and the left ventricular end systolic pressure. The parameters for diastolic function were the left ventricular end diastolic pressure and the end diastolic pressure-volume relationship (EDPVR). Cardiac tissue was analysed by ELISA for the protein content of the cytokines TNF-alpha, IL6, IL1-beta, and TGF-beta1. Phosphorylation of MAPK p38 was analysed by western blot, and the total cardiac collagen content was analysed by Sirius red staining. RESULTS: Left ventricular dysfunction was documented by impaired ESPVR and EDPVR. Cardiac cytokine levels and cardiac fibrosis were increased in diabetic animals compared to controls. Treatment with the p38 inhibitor normalised cardiac cytokine levels and improved systolic function, but did not change cardiac fibrosis and diastolic dysfunction compared to placebo. CONCLUSIONS/INTERPRETATION: Pharmacological inhibition of p38 MAPK prevents cardiac inflammation and attenuates left ventricular dysfunction in diabetic cardiomyopathy.

[Case report of Ito-syndrome associated with congenital hemihypertrophy from the orthopaedic point of view]. / Fallbericht über das Ito-Syndrom mit angeborener Hemihypertrophie aus orthopädischer Sicht.

We report on the orthopaedic treatment of a patient with the very rare Ito syndrome and congenital hemihypertrophy. The leading symptom is the lamellar depigmentation of the skin for which it is synonymously called incontinantia pigmenti acromians. Further anomalies are found in the central nervous system, as well as the ocular and the musculoskeletal systems. The treatment of the hemihypertrophy and the coexistent dysplasia of the hip with a combination of intertrochanteric shortening osteotomy and a triple osteotomy are specified and further methods are discussed.

[Limited efficacy of opioids in chronic musculoskeletal pain]. / Eingeschränkte Wirksamkeit von Opioiden bei chronischen muskuloskelettalen Schmerzen. Eine Ursachenanalyse.

After excluding malignant disease in 21 patients with unremitting strong pain of the musculoskeletal system despite long-term opioid medication, the opioids were withdrawn to search for reasons of the limited effectiveness of the opioids. The opioid withdrawal was integrated in multimodal pain coping therapy. Besides the somatic diagnoses, pain-relevant psychosomatic diagnoses were evaluated with the structured clinical interview for DSM-IV (SCID). At the time of admission and discharge pain medication, physical functions, mood, and pain intensity were recorded. In the SCID interview, all patients were diagnosed with a relevant comorbid psychiatric condition (pain disorder, anxiety, depression). Despite reduction of the opioid medication, there was no increase of pain, but an improvement of the physical functions. In patients with chronic pain of the musculoskeletal system and limited effectiveness of opioid medication, psychosomatic comorbidities should be evaluated. Instead of continued and increased opioid medication, pain coping strategies and opioid withdrawal should be tested.

[Bronchial rupture combined with luxation fracture of the thoracic spine following direct trauma]. / Bronchusruptur kombiniert mit Brustwirbelsäulenluxationsfraktur durch direktes Trauma.

Tracheobronchial injuries in blunt thoracic trauma are very rare (incidence: under 1%), with potentially devastating consequences. Appropriate pre-, intra-, and postoperative management is mandatory to ensure the patient's survival and maintain lung function. We report the case of a 62-year-old male patient hit by a tree over the chest while cutting down trees, suffering a rupture of the right bronchus and a tear of the trachea combined with a luxation fracture of the thoracic spine between Th2 and Th3 (without neurological deficit). With immediate suture of the torn bronchus and trachea and stabilization of the spine fracture on the following day, we achieved a successful outcome in this patient. To our knowledge, this is the first description in the literature of the combination of both injuries.

The virtual articulator: development and evaluation.

The accuracy, reliability, and reproducibility of the virtual articulator as applied in functional diagnostics are examined. Its potential in planning and simulating orthodontic treatment is also explored.

Polymerization shrinkage-strain and microleakage in dentin-bordered cavities of chemically and light-cured restorative materials.

OBJECTIVES: The aim of this study was to evaluate in vitro the relationship between polymerization shrinkage and microleakage in dentin-bordered restorations. METHODS: Four light-cured restorative materials in combination with their respective dental bonding agents (DBA) were investigated: Tetric Ceram/Syntac classic (Vivadent), Solitaire/Gluma Solid bond (Heraeus Kulzer), Definite/Etch & Prime 3.0 (Degussa), Solitaire 2/Gluma Solid bond (Heraeus Kulzer). The chemically cured resin Degufill sc microhybrid (Degussa) in combination with ART Bond (Coltène) was also included. Polymerization shrinkage of the restorative materials was measured using three different methods (dilatometer, linometer, buoyancy method) and analyzed with ANOVA. For the determination of microleakage, caries-free human molars were embedded in acrylic resin and subsequently abraded with a wet abrasion machine to produce four level dentin surfaces. One hundred sixty cavities (3 mm diameter/1.5 mm deep) were randomly assigned to four groups of equal size. The groups were restored without (group 1 and 2) and with DBA (group 3 and 4), and either not subjected (group 1 and 3) or subjected (group 2 and 4) to 2000 cycles from 5-55 degrees C. Each group was further divided into five material subgroups of eight cavities each. Microleakage was determined using a dye penetration test assessed at depths of 200, 400 and 600 microm into the fillings. Data were analyzed with the Kruskal-Wallis and the Mann-Whitney test. RESULTS: All three methods of measuring polymerization shrinkage (PS) generated the same, statistically secured ranking for the four light-cured restorative materials: PS Definite < PS Tetric Ceram < PS Solitaire 2 < PS Solitaire. In the microleakage study, only a few statistically significant differences were observed. Etch & Prime 3.0/Definite in group 3 and Solid Bond/Solitaire 2 in group 4 tended to exhibit the least microleakage. Correlation coefficients between aggregated shrinkage and microleakage data were 0.3 for group 3 and -0.2 for group 4. SIGNIFICANCE: The results do not suggest any correlation between polymerization shrinkage and microleakage in dentin of direct adhesive restorations.