Protein and gene expression analyses in bone marrow stem cells mediated restoration of myocardium after ischemic insult

PhDMyocardial Infarction (MI) is caused by occlusion of the coronary artery following atheromatous plaque rupture, the subsequent ischemia in the myocardium leads to myocyte necrosis unless treated quickly. Bone marrow derived stem cell treatment is a promising therapy for improving the outcome of patients with MI. The aim of this thesis was to study myocardial protein and gene expression changes in a rat ischemia/reperfusion (I/R) model in order to look for potential repair mechanisms of the myocardium triggered by endogenous bone marrow mononuclear cells (BMMNCs). Rat myocardial samples were obtained from three experimental groups: one group had a sham operation, the other two groups had undergone myocardial I/R injury induced by left anterior descending (LAD) coronary artery ligation followed by treatment with either a BMMNC preparation or PBS. Comparative proteomic analyses were carried out using 2D electrophoresis; differentially expressed proteins were identified using LC-MS/MS. Western blotting was used to confirm the most significant findings including expression of 14-3-3 epsilon protein. Global comparative gene expression profiling was performed using Illumina RatRef12 BeadChips and QPCR was used to validate the top results. Bioinformatic tools were used to assess the biology of the differentially expressed genes and proteins. Thirty-seven proteins were found to be differentially expressed in I/R injury compared to sham. These were primarily sarcomeric, energy production or stress response proteins and most were down-regulated. Expression levels were ‘corrected’ by BMMNC treatment for many of these proteins. Over 1500 genes were affected by I/R injury, 20 were affected by BMMNC treatment, and many of these were related to inflammation and apoptosis signalling and responses. The 14-3-3 epsilon protein was chosen for follow-up work as it presented as a good candidate for mechanistic involvement. This protein has many roles including interactions with the proapoptotic BCL2-associated agonist of cell death (Bad) protein. Western blotting was used to look at Bad expression and found it to be significantly increased in the Page 3 treatment group, although I could not reliably measure the expression of phosphorylated (serine 136) form of Bad. A preliminary pull-down assay was performed to look for binding partners of 14-3-3 epsilon. Two ATP synthase subunits, one of which is known to bind 14-3-3 epsilon, a protein involved in fatty acid β-oxidation and a protein of unknown function were found to bind. Further work will be required to follow up these findings and ascertain the exact role of 14-3- 3 epsilon in cardioprotection. In summary, my data supports the power of profiling methods to derive new candidates for a role in repair mechanisms in this therapeutic model

Global public health training in the UK: preparing for the future.

BACKGROUND: Many major public health issues today are not confined by national boundaries. However, the global public health workforce appears unprepared to confront the challenges posed by globalization. We therefore sought to investigate whether the current UK public health training programme adequately prepares its graduates to operate in a globalized world. METHODS: We used mixed methods involving an online cross-sectional survey of UK public health trainees on the international content of the Faculty of Public Health's written examination, a qualitative review of the Faculty's 2007 training curriculum and a questionnaire survey of all training deaneries in the UK. RESULTS: We found that global health issues are not addressed by the current training curriculum or in the written examination despite trainee interest for this. Many of the deaneries were also unreceptive to international placements. CONCLUSIONS: Despite the recognized educational legitimacy of global health placements and the favourable UK policy context, the opportunities and international content of public health training remain limited. In order to retain its position as a leader in the field of public health, the UK needs to adapt its training programme to better reflect today's challenges

Reef-fidelity and migration of tiger sharks, Galeocerdo cuvier, across the Coral Sea

© The Author(s), 2014. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 9 (2014): e83249, doi:10.1371/journal.pone.0083249.Knowledge of the habitat use and migration patterns of large sharks is important for assessing the effectiveness of large predator Marine Protected Areas (MPAs), vulnerability to fisheries and environmental influences, and management of shark–human interactions. Here we compare movement, reef-fidelity, and ocean migration for tiger sharks, Galeocerdo cuvier, across the Coral Sea, with an emphasis on New Caledonia. Thirty-three tiger sharks (1.54 to 3.9 m total length) were tagged with passive acoustic transmitters and their localised movements monitored on receiver arrays in New Caledonia, the Chesterfield and Lord Howe Islands in the Coral Sea, and the east coast of Queensland, Australia. Satellite tags were also used to determine habitat use and movements among habitats across the Coral Sea. Sub-adults and one male adult tiger shark displayed year-round residency in the Chesterfields with two females tagged in the Chesterfields and detected on the Great Barrier Reef, Australia, after 591 and 842 days respectively. In coastal barrier reefs, tiger sharks were transient at acoustic arrays and each individual demonstrated a unique pattern of occurrence. From 2009 to 2013, fourteen sharks with satellite and acoustic tags undertook wide-ranging movements up to 1114 km across the Coral Sea with eight detected back on acoustic arrays up to 405 days after being tagged. Tiger sharks dove 1136 m and utilised three-dimensional activity spaces averaged at 2360 km3. The Chesterfield Islands appear to be important habitat for sub-adults and adult male tiger sharks. Management strategies need to consider the wide-ranging movements of large (sub-adult and adult) male and female tiger sharks at the individual level, whereas fidelity to specific coastal reefs may be consistent across groups of individuals. Coastal barrier reef MPAs, however, only afford brief protection for large tiger sharks, therefore determining the importance of other oceanic Coral Sea reefs should be a priority for future research.Funding was provided by the the Agence Francaise de Développement (http://www.afd.fr), French Pacific Fund, the CRISP program (www.crisponline.info) and QLD Fisheries

The Effects of Physical Activity on Learning Behaviors in Elementary School Children: A Randomized Controlled Trial

This work is licensed under a Creative Commons Attribution 4.0 International License.Research in education and developmental psychology indicates that behavioral engagement in learning is a critical predictor of children’s academic success. In an effort to improve academic achievement, school administrators are continually in search of methods to increase behavioral engagement. Previous research has indicated that classroom-based physical activity (PA) lessons have a positive impact on academic achievement. However, little research has been done in assessing the impact of such interventions on the behavioral engagement of students with learning behavior difficulties. This study assesses the impact of classroom-based PA on teacher-rated classroom behaviors of students with identified learning behavior difficulties. Two schools (one intervention, one control) participating in a larger, cluster-randomized trial provided scores on a teacher-administered classroom behavior scale. This scale was used to collect information on 15 characteristics identified as being essential to behavioral engagement. Participants included male and female students in second and third grade classrooms who were identified by their classroom teacher and school counselor as having difficulties with learning behaviors. Mixed linear modeling for repeated measures was used to examine the changes over time in the classroom behavior scores. The intervention group showed significant improvement over time in classroom behavior while the control group showed no change or a slight degradation over time (i.e., group × time interaction, F[2132] = 4.52, p = 0.01). Schools must meet the diverse needs of students today, including those who exhibit less than optimal learning behaviors. Combined with the evidence that PA is linked to several health and cognitive-behavior benefits, providing classroom-based PA that is incorporated within the curriculum provides common ground for all students to participate. It is a potential solution to increasing behavioral engagement, and in turn stimulating and enhancing learning

LINE Retrotransposon RNA Is an Essential Structural and Functional Epigenetic Component of a Core Neocentromeric Chromatin

We have previously identified and characterized the phenomenon of ectopic human centromeres, known as neocentromeres. Human neocentromeres form epigenetically at euchromatic chromosomal sites and are structurally and functionally similar to normal human centromeres. Recent studies have indicated that neocentromere formation provides a major mechanism for centromere repositioning, karyotype evolution, and speciation. Using a marker chromosome mardel(10) containing a neocentromere formed at the normal chromosomal 10q25 region, we have previously mapped a 330-kb CENP-A–binding domain and described an increased prevalence of L1 retrotransposons in the underlying DNA sequences of the CENP-A–binding clusters. Here, we investigated the potential role of the L1 retrotransposons in the regulation of neocentromere activity. Determination of the transcriptional activity of a panel of full-length L1s (FL-L1s) across a 6-Mb region spanning the 10q25 neocentromere chromatin identified one of the FL-L1 retrotransposons, designated FL-L1b and residing centrally within the CENP-A–binding clusters, to be transcriptionally active. We demonstrated the direct incorporation of the FL-L1b RNA transcripts into the CENP-A–associated chromatin. RNAi-mediated knockdown of the FL-L1b RNA transcripts led to a reduction in CENP-A binding and an impaired mitotic function of the 10q25 neocentromere. These results indicate that LINE retrotransposon RNA is a previously undescribed essential structural and functional component of the neocentromeric chromatin and that retrotransposable elements may serve as a critical epigenetic determinant in the chromatin remodelling events leading to neocentromere formation

Clean birth and postnatal care practices to reduce neonatal deaths from sepsis and tetanus: a systematic review and Delphi estimation of mortality effect

BACKGROUND: Annually over 520,000 newborns die from neonatal sepsis, and 60,000 more from tetanus. Estimates of the effect of clean birth and postnatal care practices are required for evidence-based program planning. OBJECTIVE: To review the evidence for clean birth and postnatal care practices and estimate the effect on neonatal mortality from sepsis and tetanus for the Lives Saved Tool (LiST). METHODS: We conducted a systematic review of multiple databases. Data were abstracted into standard tables and assessed by GRADE criteria. Where appropriate, meta-analyses were undertaken. For interventions with low quality evidence but a strong GRADE recommendation, a Delphi process was conducted. RESULTS: Low quality evidence supports a reduction in all-cause neonatal mortality (19% (95% c.i. 1-34%)), cord infection (30% (95% c.i. 20-39%)) and neonatal tetanus (49% (95% c.i. 35-62%)) with birth attendant handwashing. Very low quality evidence supports a reduction in neonatal tetanus mortality with a clean birth surface (93% (95% c.i. 77-100%)) and no relationship between a clean perineum and tetanus. Low quality evidence supports a reduction of neonatal tetanus with facility birth (68% (95% c.i. 47-88%). No relationship was found between birth place and cord infections or sepsis mortality. For postnatal clean practices, all-cause mortality is reduced with chlorhexidine cord applications in the first 24 hours of life (34% (95% c.i. 5-54%, moderate quality evidence) and antimicrobial cord applications (63% (95% c.i. 41-86%, low quality evidence). One study of postnatal maternal handwashing reported reductions in all-cause mortality (44% (95% c.i. 18-62%)) and cord infection ((24% (95% c.i. 5-40%)).Given the low quality of evidence, a Delphi expert opinion process was undertaken. Thirty experts reached consensus regarding reduction of neonatal sepsis deaths by clean birth practices at home (15% (IQR 10-20)) or in a facility (27% IQR 24-36)), and by clean postnatal care practices (40% (IQR 25-50)). The panel estimated that neonatal tetanus mortality was reduced by clean birth practices at home (30% (IQR(20-30)), or in a facility (38% (IQR 34-40)), and by clean postnatal care practices (40% (IQR 30-50)). CONCLUSION: According to expert opinion, clean birth and particularly postnatal care practices are effective in reducing neonatal mortality from sepsis and tetanus. Further research is required regarding optimal implementation strategies

Maximising HIV prevention by balancing the opportunities of today with the promises of tomorrow: a modelling study

SummaryBackgroundMany ways of preventing HIV infection have been proposed and more are being developed. We sought to construct a strategic approach to HIV prevention that would use limited resources to achieve the greatest possible prevention impact through the use of interventions available today and in the coming years.MethodsWe developed a deterministic compartmental model of heterosexual HIV transmission in South Africa and formed assumptions about the costs and effects of a range of interventions, encompassing the further scale-up of existing interventions (promoting condom use, male circumcision, early antiretroviral therapy [ART] initiation for all [including increased HIV testing and counselling activities], and oral pre-exposure prophylaxis [PrEP]), the introduction of new interventions in the medium term (offering intravaginal rings, long-acting injectable antiretroviral drugs) and long term (vaccine, broadly neutralising antibodies [bNAbs]). We examined how available resources could be allocated across these interventions to achieve maximum impact, and assessed how this would be affected by the failure of the interventions to be developed or scaled up.FindingsIf all interventions are available, the optimum mix would place great emphasis on the following: scale-up of male circumcision and early ART initiation with outreach testing, as these are available immediately and assumed to be low cost and highly efficacious; intravaginal rings targeted to sex workers; and vaccines, as these can achieve a large effect if scaled up even if imperfectly efficacious. The optimum mix would rely less on longer term developments, such as long-acting antiretroviral drugs and bNAbs, unless the costs of these reduced. However, if impossible to scale up existing interventions to the extent assumed, emphasis on oral PrEP, intravaginal rings, and long-acting antiretroviral drugs would increase. The long-term effect on the epidemic is most affected by scale-up of existing interventions and the successful development of a vaccine.InterpretationWith current information, a strategic approach in which limited resources are used to maximise prevention impact would focus on strengthening the scale-up of existing interventions, while pursuing a workable vaccine and developing other approaches that can be used if further scale-up of existing interventions is limited.FundingBill & Melinda Gates Foundation

Multi-segment spine kinematics: Relationship with dance training and low back pain

Background: Spine posture, range of motion (ROM) and movement asymmetry can contribute to low back pain (LBP). These variables may have greater impact in populations required to perform repetitive spine movements, such as dancers; however, there is limited evidence to support this. Research question: What is the influence of dance and LBP on spinal kinematics? Methods: In this cross-sectional study, multi-segment spinal kinematics were examined in 60 female participants, including dancers (n = 21) and non-dancers (n = 39) with LBP (n = 33) and without LBP (n = 27). A nine-camera motion analysis system sampling at 100 Hz was used to assess standing posture, as well as ROM and movement asymmetry for side bend and trunk rotation tasks. A two-way ANOVA was performed for each of the outcome variables to detect any differences between dancers and non-dancers, or individuals with and without LBP. Results: Compared to non-dancers, dancers displayed a flatter upper lumbar angle when standing (p  0.05) or movement asymmetry (p > 0.05). There was no main effect for LBP symptoms on any kinematic measures, and no interaction effect for dance group and LBP on spinal kinematics (p > 0.05). Significance: Female dancers displayed a flatter spine posture and increased spine ROM compared to non-dancers for a select number of spine segments and movement tasks. However, the overall number of differences was small, and no relationship was observed between LBP and spinal kinematics. This suggests that these simple, static posture, ROM, and asymmetry measures often used in clinical practice can provide only limited generalisable information about the impact of dance or LBP on spinal kinematics