Extended 2D myotube culture recapitulates postnatal fibre type plasticity

Background: The traditional problems of performing skeletal muscle cell cultures derived from mammalian or avian species are limited myotube differentiation, and transient myotube persistence which greatly restricts the ability of myotubes to undergo phenotypic maturation. We report here on a major technical breakthrough in the establishment of a simple and effective method of extended porcine myotube cultures (beyond 50 days) in two-dimension (2D) that recapitulates key features of postnatal fibre types. Results: Primary porcine muscle satellite cells (myoblasts) were isolated from the longissimus dorsi of 4 to 6 weeks old pigs for 2D cultures to optimise myotube formation, improve surface adherence and characterise myotube maturation. Over 95 % of isolated cells were myoblasts as evidenced by the expression of Pax3 and Pax7. Our relatively simple approach, based on modifications of existing surface coating reagents (Maxgel), and of proliferation and differentiation (Ultroser G) media, typically achieved by 5 days of differentiation fusion index of around 80 % manifested in an abundance of discrete myosin heavy chain (MyHC) slow and fast myotubes. There was little deterioration in myotube viability over 50 days, and the efficiency of myotube formation was maintained over seven myoblast passages. Regular spontaneous contractions of myotubes were frequently observed throughout culture. Myotubes in extended cultures were able to undergo phenotypic adaptation in response to different culture media, including the adoption of a dominant postnatal phenotype of fast-glycolytic MyHC 2x and 2b expression by about day 20 of differentiation. Furthermore, fast-glycolytic myotubes coincided with enhanced expression of the putative porcine long intergenic non-coding RNA (linc-MYH), which has recently been shown to be a key coordinator of MyHC 2b expression in vivo. Conclusions: Our revised culture protocol allows the efficient differentiation and fusion of porcine myoblasts into myotubes and their prolonged adherence to the culture surface. Furthermore, we are able to recapitulate in 2D the maturation process of myotubes to resemble postnatal fibre types which represent a major technical advance in opening access to the in vitro study of coordinated postnatal muscle gene expression

Developing theory-informed behaviour change interventions to implement evidence into practice : a systematic approach using the Theoretical Domains Framework

PMID: 22531013 [PubMed - indexed for MEDLINE] PMCID: PMC3443064 Free PMC ArticlePeer reviewedPublisher PD

Gathering opinion leader data for a tailored implementation intervention in secondary healthcare: a randomised trial

Background: Health professionals’ behaviour is a key component in compliance with evidence-based recommendations. Opinion leaders are an oft-used method of influencing such behaviours in implementation studies, but reliably and cost effectively identifying them is not straightforward. Survey and questionnaire based data collection methods have potential and carefully chosen items can – in theory – both aid identification of opinion leaders and help in the design of an implementation strategy itself. This study compares two methods of identifying opinion leaders for behaviour-change interventions. Methods: Healthcare professionals working in a single UK mental health NHS Foundation Trust were randomly allocated to one of two questionnaires. The first, slightly longer questionnaire, asked for multiple nominations of opinion leaders, with specific information about the nature of the relationship with each nominee. The second, shorter version, asked simply for a list of named “champions” but no more additional information. We compared, using Chi Square statistics, both the questionnaire response rates and the number of health professionals likely to be influenced by the opinion leaders (i.e. the “coverage” rates) for both questionnaire conditions. Results: Both questionnaire versions had low response rates: only 15% of health professionals named colleagues in the longer questionnaire and 13% in the shorter version. The opinion leaders identified by both methods had a low number of contacts (range of coverage, 2–6 each). There were no significant differences in response rates or coverage between the two identification methods. Conclusions: The low response and population coverage rates for both questionnaire versions suggest that alternative methods of identifying opinion leaders for implementation studies may be more effective. Future research should seek to identify and evaluate alternative, non-questionnaire based, methods of identifying opinion leaders in order to maximise their potential in organisational behaviour change interventions

Free calcium concentration, calpain-2 activity, and final product tenderness of electrically stimulated beef

The objective of this study was to evaluate the effect of timing of electrical stimulation on free calcium concentration, calpain-2 activity, Warner-Bratzler shear force (WBSF), and consumer sensory analysis. Twenty-three beef steers were harvested and stimulated (S) using extra-low voltage or not stimulated (NS), at exsanguination and/or 1 h postmortem, resulting in 4 different stimulation treatments: NS-NS, NS-S, S-NS, or S-S. Samples were removed from the longissimus lumborum (LL) and semimembranosus (SM) for free calcium and calpain-2 analysis on days 1, 4, and 14 postmortem. WBSF and sensory analysis steaks were removed on day 4 and frozen (4 d) or aged to 14 d postmortem. Data were analyzed using the mixed model or generalized linear mixed model procedure of SAS (SAS Institute, Inc., Cary, NC), with significance determined at P < 0.05. There was a tendency for an aging-period-by-stimulation-treatment interaction for LL free calcium concentration (P = 0.05), and there was a significant difference between aging periods (P < 0.01). No difference was observed in free calcium concentration in the SM between stimulation treatments (P = 0.44); aging, however, significantly increased SM free calcium concentration (P < 0.01). Stimulation did not impact native calpain-2 activity in the LL (P = 0.71) or SM (P = 0.89). Stimulation treatment did not improve tenderness values for WBSF analysis for the LL (P = 0.69) or SM (P = 0.61) or consumer sensory analysis in the LL (P = 0.56) or SM (P = 0.36). A longer aging period tended to increase calpain-2 activity in the SM (P = 0.08), improve WBSF in the LL (P = 0.09), and significantly improve consumer tenderness scores in the SM (P < 0.01). In conclusion, the timing of electrical stimulation utilized in the current study tended to influence free calcium concentration in the LL but did not influence calpain-2 activity or beef tenderness. Aging, however, improved tenderness.This work is/was supported by the USDA National Institute of Food and Agriculture, Hatch Project W4177 Enhancing the Competitiveness and Value of U.S. Beef. Support was also received from the Idaho Experiment Station

Face validity of the Functional Assessment of Cancer Therapy-Breast Symptom Index (FACT- B) into formal Arabic

Background: Breast cancer affects over one million women annually and is the most common global malignancy among women. Extensive improvements have taken place in the management of breast cancer in recent years and a higher percentage of women are cured from this disease. A proper assessment of the quality of life of women with breast cancer is an essential component in disease management. The Functional Assessment of Cancer Therapy- Breast Symptom Index has been commonly used and well-validated among English speaking populations as well as other populations. To date, no formal translation and evaluation of the Functional Assessment of Cancer Therapy-Breast System Index exists in Arabic. Therefore, this study intends to translate, adapt and face-validate the Functional Assessment of Cancer Therapy-Breast System Index into Arabic, specifically in the context of the Lebanese culture. Methods: We conducted forward and backward translation in Arabic, combined with face validity by clinicians. This was followed by pre-testing to ensure the instrument's adequacy and cultural sensitivity conducted by the administration of face-to-face interviews with individual breast cancer patients (n=33) and two focus groups (4 women/group) to evaluate the relevance and appropriateness of each item and words used in the questionnaire. Results: Study results reinforced the value of the Arabic translated version of the Functional Assessment of Cancer Therapy-Breast System Index in capturing the quality of life of women with breast cancer in Lebanon. Conclusion: The instrument was perceived to be adequate, appropriate for use, culturally sensitive, simple as well as exhaustive. Suggestions have been made to enrich the instruments' ability to incorporate other quality of life dimensions not captured, as well to enhance the cultural specificity of the Functional Assessment of Cancer Therapy-Breast System Index, when administered among Lebanese women diagnosed with breast cancer who are specifically beyond therapy

Impact of Protracted War Crisis on Dental Students: A Comparative Multicountry Cross-sectional Study

BACKGROUND: The impact of conflict and war crisis on dental students is poorly understood. Given the prolonged conflicts and political instability in the Arabic-speaking countries, it is crucial to investigate the effect of these conditions on dental students. This study aimed to assess the impact of protracted war on dental students by comparing the personal, university, and wider context challenges they face across war-affected and unaffected countries. METHODS: A cross-sectional study was conducted including a convenience sample of dental students from 13 universities in 12 Arabic-speaking countries. Respondents were those at entry and exit points of their undergraduate dental training. A self-administered paper questionnaire collected anonymized data on sociodemographics, and personal, university, and wider context challenges that students were facing. Multivariable Poisson regression analyses were carried out. RESULTS: The overall response rate was 64.8%. The mean age was 21.2 (standard deviation = 2.1) years, with 68% of participants being female. After adjusting for age and sex, dental students in Arabic-speaking countries affected by protracted war crisis were significantly more likely to report wider context challenges compared to their counterparts in unaffected countries (n = 2448; beta = 1.12; 95% confidence interval: 1.10-1.13; P < 0.001). DISCUSSION: Dental students in Arabic-speaking countries affected by protracted war crisis were more likely to suffer from wider context challenges such as difficulties in attendance due to the deterioration of security and lack of flexibility of teaching time to accommodate the different circumstances induced by the war crisis. Supporting dental students in areas affected by protracted war crises is needed and may include developing online dental education programs

Validity and reliability of the Arabic version of Activities of Daily Living (ADL)

Background. The Activity of Daily Living (ADL) is an instrument that screens elderly respondents for physical functioning and assesses whether they are dependent or independent in their daily activities. This study demonstrates a translation procedure and obtains the reliability and validity of a translated, Arabic ADL. Methods. The ADL was translated to Arabic through a forward translation method followed by a committee-consensual approach. The ADL and the Arabic Mini-Mental State Examination (AMMSE) were administered to an opportunistic sample of 354 Lebanese elderly living in nursing homes who did not have dementia. Results. Reliability split half measures, sensitivity, and negative predictive values were high across all dimensions of the ADL with the exception of feeding. There were non-significant differences on the scored ADL between the three age groups: young age, middle age and older old. In addition, a non-significant difference was found on the scored ADL between the high and low AMMSE scores. Conclusion. Overall, the translated ADL was consistent and valid measure for assessing daily activities in elderly nursing home residents. As it is quick and easy to use, the ADL in Arabic could help caregivers and doctors to prescribe appropriate physical exercise for elderly Arabic speaking patients

Impedance Control of a Pneumatic Muscles-Driven Ankle Rehabilitation Robot

Pneumatic muscle is a new type of flexible actuator with advantages in terms of light weight, large output power/weight ratio, good security, low price and clean. In this paper, an ankle rehabilitation robot with two degrees of freedom driven by pneumatic muscle is studied. The force control method with an impedance controller in outer loop and a position inner loop is proposed. The demand of rehabilitation torque is ensured through tracking forces of three pneumatic muscle actuators. In the simulation, the constant force and variable force are tracked with error less than 10 N. In the experiment, the force control method also achieved satisfactory results, which provides a good support for the application of the robot in the ankle rehabilitation

Maternal and Child Oral Health Interventions in Middle East and North Africa Regions:a rapid review

Objectives: To conduct a rapid review to identify any maternal and/or child oral health interventions implemented and/or tested in Middle East and North Africa (MENA) countries generally, and Lebanon, Palestine and Syria specifically, and to compile information on the relative effectiveness of these interventions. Methods: A systematic search was conducted for primary and secondary literature indexed in five online databases, and the websites of the World Health Organisation (WHO), the International Union for Health Promotion and Education (IUHPE), the United Nations Children’s Fund (UNICEF), United Nations Relief and Works Agency for Palestine Refugees in the Near East (UNRWA), and the United Nations Refugee Agency (UNHCR). Results: Two independent researchers reviewed 1,180 records from the online databases, and 3,120 reports from the WHO, UNRWA, UNICEF and UNHCR. Four unique studies were included and conducted in Syria, Saudi Arabia and Iran. No systematic reviews were found for targeted interventions in MENA regions. However, interventions using fluoridated toothpaste (Syria), preventive treatment and fluoridated chewing gum (Saudi Arabia), and oral health education with oral health reminders (Iran) were significantly effective in reducing early child caries (ECC) experience. In Syria and Iran, mother and child oral health promotions integrated into ongoing vaccination programmes were effective in reducing ECC. These interventions formed part of WHO and Ministry of Health programmes. Conclusion: Further investigation is essential to verify the effectiveness of incorporating multi-disciplinary, theory-driven oral health interventions into ongoing WHO maternal and child health programmes in MENA countries to assist in promoting oral health and wellbeing

Standards for the care of people with cystic fibrosis; establishing and maintaining health

This is the second in a series of four papers updating the European Cystic Fibrosis Society (ECFS) standards for the care of people with CF. This paper focuses on establishing and maintaining health. The guidance is produced using an evidence-based framework and with wide stakeholder engagement, including people from the CF community. Authors provided a narrative description of their topic and statements, which were more directive. These statements were reviewed by a Delphi exercise, achieving good levels of agreement from a wide group for all statements. This guidance reinforces the importance of a multi-disciplinary CF team, but also describes developing models of care including virtual consultations. The framework for health is reinforced, including the need for a physically active lifestyle and the strict avoidance of all recreational inhalations, including e-cigarettes. Progress with cystic fibrosis transmembrane conductance regulator (CFTR) modulator therapy is reviewed, including emerging adverse events and advice for dose reduction and interruption. This paper contains guidance that is pertinent to all people with CF regardless of age and eligibility for and access to modulator therapy