Investigation of the role of long non-coding RNAs in oncogene induced cellular senescence

Cellular senescence is a stable arrest of cell proliferation induced by several factors such as activated oncogenes, oxidative stress and shortening of telomeres. Senescence acts as a tumour suppression mechanism to halt the progression of cancer. However, senescence may also impact negatively upon tissue regeneration, thus contributing to the effects of ageing. The eukaryotic genome is controlled by various modes of transcriptional and translational regulation. Focus has therefore centred on the role of long non- coding RNAs (lncRNAs) in regulating the genome. Accordingly, understanding how lncRNAs function to regulate the senescent genome is integral to improving our knowledge and understanding of tumour suppression and ageing. Within this study, I set out to investigate the expression of lncRNAs’ expression within models of senescence. Through a custom expression array, I have shown that expression of multiple different lncRNAs is up-regulated and down regulated in IMR90 replicative senescent fibroblasts and oncogene-induced senescent melanocytes. LncRNA expression was determined to be specific to stable senescence-associated cell arrest and predominantly within the nucleus of senescent cells. In order to examine the function of lncRNA expression in senescence, I selected lncRNA transcript ENST0000430998 (lncRNA_98) to focus my investigations upon. LncRNA_98 was robustly upregulated within multiple models of senescence and efficiently depleted using anti-sense oligonucleotide technology. Characterisation and unbiased RNA-sequencing of lncRNA_98 deficient senescent cells highlighted a list of genes that are regulated by lncRNA_98 expression in senescent cells and may regulate aspects of the senescence program. Specifically, the formation of SAHF was impeded upon depletion of lncRNA_98 expression and levels of total pRB protein expression severely decreased. Validation and recapitulation of consequences of pRB depletion was confirmed through lncRNA_98 knock-out cells generated using CRISPR technology. Surprisingly, inhibition of ATM kinase functions permitted the restoration of pRB protein levels within lncRNA_98 deficient cells. I propose that lncRNA_98 antagonizes the ability of ATM kinase to downregulate pRB expression at a post-transcriptional level, thereby potentiating senescence. Furthermore, lncRNA expression was detected within fibroblasts of old individuals and visualised within senescent melanocytes in human benign nevi, a barrier to melanoma progression. Conversely, mining of 337 TCGA primary melanoma data sets highlighted that the lncRNA_98 gene and its expression was lost from a significant proportion of melanoma samples, consistent with lncRNA_98 having a tumour suppressor functions. The data presented in this study illustrates that lncRNA_98 expression has a regulatory role over pRB expression in senescence and may regulate aspects of tumourigenesis and ageing.

Long distance coupling of a quantum mechanical oscillator to the internal states of an atomic ensemble

We propose and investigate a hybrid optomechanical system consisting of a micro-mechanical oscillator coupled to the internal states of a distant ensemble of atoms. The interaction between the systems is mediated by a light field which allows the coupling of the two systems in a modular way over long distances. Coupling to internal degrees of freedom of atoms opens up the possibility to employ high-frequency mechanical resonators in the MHz to GHz regime, such as optomechanical crystal structures, and to benefit from the rich toolbox of quantum control over internal atomic states. Previous schemes involving atomic motional states are rather limited in both of these aspects. We derive a full quantum model for the effective coupling including the main sources of decoherence. As an application we show that sympathetic ground-state cooling and strong coupling between the two systems is possible.

Evaluation use within a humanitarian non-governmental organization’s health care user-fee exemption program in West Africa

L’évaluation de l’action humanitaire (ÉAH) est un outil valorisé pour soutenir l’imputabilité, la transparence et l’efficience de programmes humanitaires contribuant à diminuer les inéquités et à promouvoir la santé mondiale. L’EAH est incontournable pour les parties prenantes de programme, les bailleurs de fonds, décideurs et intervenants souhaitant intégrer les données probantes aux pratiques et à la prise de décisions. Cependant, l’utilisation de l’évaluation (UÉ) reste incertaine, l’ÉAH étant fréquemment menée, mais inutilisé. Aussi, les conditions influençant l’UÉ varient selon les contextes et leur présence et applicabilité au sein d’organisations non-gouvernementales (ONG) humanitaires restent peu documentées. Les évaluateurs, parties prenantes et décideurs en contexte humanitaire souhaitant assurer l’UÉ pérenne détiennent peu de repères puisque rares sont les études examinant l’UÉ et ses conditions à long terme. La présente thèse tend à clarifier ces enjeux en documentant sur une période de deux ans l’UÉ et les conditions qui la détermine, au sein d’une stratégie d’évaluation intégrée au programme d’exemption de paiement des soins de santé d’une ONG humanitaire. L’objectif de ce programme est de faciliter l’accès à la santé aux mères, aux enfants de moins de cinq ans et aux indigents de districts sanitaires au Niger et au Burkina Faso, régions du Sahel où des crises alimentaires et économiques ont engendré des taux élevés de malnutrition, de morbidité et de mortalité. Une première évaluation du programme d’exemption au Niger a mené au développement de la stratégie d’évaluation intégrée à ce même programme au Burkina Faso. La thèse se compose de trois articles. Le premier présente une étude d’évaluabilité, étape préliminaire à la thèse et permettant de juger de sa faisabilité. Les résultats démontrent une logique cohérente et plausible de la stratégie d’évaluation, l’accessibilité de données et l’utilité d’étudier l’UÉ par l’ONG. Le second article documente l’UÉ des parties prenantes de la stratégie et comment celle-ci servit le programme d’exemption. L’utilisation des résultats fut instrumentale, conceptuelle et persuasive, alors que l’utilisation des processus ne fut qu’instrumentale et conceptuelle. Le troisième article documente les conditions qui, selon les parties prenantes, ont progressivement influencé l’UÉ. L’attitude des utilisateurs, les relations et communications interpersonnelles et l’habileté des évaluateurs à mener et à partager les connaissances adaptées aux besoins des utilisateurs furent les conditions clés liées à l’UÉ. La thèse contribue à l’avancement des connaissances sur l’UÉ en milieu humanitaire et apporte des recommandations aux parties prenantes de l’ONG.

SCOR WG 142: Quality Control Procedures for Oxygen and Other Biogeochemical Sensors on Floats and Gliders. Recommendation for oxygen measurements from Argo floats, implementation of in-air-measurement routine to assure highest long-term accuracy

Recommendation for Oxygen Measurements from Argo Floats: Implementation of In-Air-Measurement Routine to Assure Highest Long-term Accuracy As Argo has entered its second decade and chemical/biological sensor technology is improving constantly, the marine biogeochemistry community is starting to embrace the successful Argo float program. An augmentation of the global float observatory, however, has to follow rather stringent constraints regarding sensor characteristics as well as data processing and quality control routines. Owing to the fairly advanced state of oxygen sensor technology and the high scientific value of oceanic oxygen measurements (Gruber et al., 2010), an expansion of the Argo core mission to routine oxygen measurements is perhaps the most mature and promising candidate (Freeland et al., 2010). In this context, SCOR Working Group 142 “Quality Control Procedures for Oxygen and Other Biogeochemical Sensors on Floats and Gliders” (www.scor-int.org/SCOR_WGs_WG142.htm) set out in 2014 to assess the current status of biogeochemical sensor technology with particular emphasis on float-readiness, develop pre- and post-deployment quality control metrics and procedures for oxygen sensors, and to disseminate procedures widely to ensure rapid adoption in the community.

Is the supply of long-term debt independent of the term premia? Evidence from Portugal

An important assumption in the statistical analysis of the financial market effects of the central bank’s large scale asset purchase program is that the "long-term debt stock variables were exogenous to term premia". We test this assumption for a small open economy in a currency union over the period 2000M3 to 2015M10, via the determinants of short- term financing relative to long-term financing. Empirical estimations indicate that the maturity composition of debt does not respond to the level of interest rate or to the term structure. These findings suggest a lower adherence to the cost minimization mandate of debt management. However, we find that volatility and relative market size respectively decrease and increase short-term financing relative to long-term financing, while it decreases with an increase in government indebtedness.

Design of maintenance structures for rural electrification with solar home systems. The case of the Moroccan program.

n decentralised rural electrification through solar home systems, private companies and promoting institutions are faced with the problem of deploying maintenance structures to operate and guarantee the service of the solar systems for long periods (ten years or more). The problems linked to decentralisation, such as the dispersion of dwellings, difficult access and maintenance needs, makes it an arduous task. This paper proposes an innovative design tool created ad hoc for photovoltaic rural electrification based on a real photovoltaic rural electrification program in Morocco as a special case study. The tool is developed from a mathematical model comprising a set of decision variables (location, transport, etc.) that must meet certain constraints and whose optimisation criterion is the minimum cost of the operation and maintenance activity assuming an established quality of service. The main output of the model is the overall cost of the maintenance structure. The best location for the local maintenance headquarters and warehouses in a given region is established, as are the number of maintenance technicians and vehicles required.

Is Spain Different? Innovation systems in the long-term development of the tourism sector. The IBEROSTAR case study.

Tourism is a highly competitive sector which is characterized by permanent transformation. Therefore, tourism firms depend largely on innovation to adapt themselves and compete, ensuring their survival and competitiveness. Despite research on innovation concludes that tourism firms are in most cases only moderately innovative, there are outstanding exceptions in specific fields and destinies. Spain, which is an unquestionable leader in tourism, constitutes one of them. In fact, Spanish tourism firms have shown a strong innovation capacity. They have improved and diversified their products thus meeting the requirements of a changing demand. This raises the need to properly understand how and why innovation takes place and who is involved in such a process. This paper aims to address these questions through the case study of one of the largest Spanish hospitality firms. The long history of Iberostar Group confirms that family firms and the networks they provide, along with environments that foster competition as was the case of the Spanish tourism industry, are important determinants of innovation particularly in a late-developed country.

Review of the Community Residential Care Facilities Program, Department of Health and Environmental Control

This review was predicated on a credible complaint alleging substantial health and safety deficiencies in the care of a resident placed in a Community Residential Care Facility (CRCF) in Kershaw County, South Carolina. Initial investigation with subject matter experts, non-profit advocacy groups, and CRCF inspection reports revealed this single incident might be a symptom of systemic health and safety deficiencies throughout DHEC’s CRCF Program, which regulates 471 CRCFs with the approximately 17,000 vulnerable clients, primarily elderly and disabled. This review’s scope and objectives were: Assess the risk of a vulnerable population of elderly and disabled citizens residing in CRCFs living in unsatisfactory health and safety conditions; Evaluate DHEC’s CRCF Program inspection process capabilities to identify and address CRCFs with unsatisfactory health and safety living conditions; Recommend opportunities to improve the CRCF Program.

Impact on hip fracture mortality after the establishment of an orthogeriatric care program in a colombian hospital

El objetivo del estudio es evaluar la mortalidad a un año en pacientes con fractura de cadera, mayores de 65 años tratados en un programa establecido de orto-geriatría. 298 se trataron de acuerdo al protocolo de orto-geriatría, se calculo la mortalidad a un año, se establecieron los predictores de mortalidad orto-geriátrico. La sobrevida anual se incremento de 80% a 89% (p = .039) durante los cuatro años de seguimiento del programa y disminuyo el riesgo de mortalidad anual postoperatorio (Hazard Ratio = 0.54, p = .049). La enfermedad cardiaca y la edad maor a 85 años fueron predictores positivos para mortalidad.

Essays in Long-run Development and Spatial Economics

This thesis tries to further our understanding for why some countries today are more prosperous than others. It establishes that part of today's observed variation in several proxies such as income or gender inequality have been determined in the distant past. Chapter one shows that 450 years of (Catholic) Portuguese colonisation had a long-lasting impact in India when it comes to education and female emancipation. Furthermore I use a historical quasi-experiment that happened 250 years ago in order to show that different outcomes have different degrees of persitence over time. Educational gaps between males and females seemingly wash out a few decades after the public provision of schools. The male biased sex-ratios on the other hand stay virtually unchanged despite governmental efforts. This provides evidence that deep rooted son preferences are much harder to overcome, suggesting that a differential approach is needed to tackle sex-selective abortion and female neglect. The second chapter proposes improvements for the execution of Spatial Regression Discontinuity Designs. These suggestions are accompanied by a full-fledged spatial statistical package written in R. Chapter three introduces a quantitative economic geography model in order to study the peculiar evolution of the European urban system on its way to the Industrial Revolution. It can explain the shift of economic gravity from the Mediterranean towards the North-Sea ("little divergence"). The framework provides novel insights on the importance of agricultural trade costs and the peculiar geography of Europe with its extended coastline and dense network of navigable rivers.

Mechanisms Predisposing Penile Fracture And Long-term Outcomes On Erectile And Voiding Functions.

Purpose. To determine the mechanisms predisposing penile fracture as well as the rate of long-term penile deformity and erectile and voiding functions. Methods. All fractures were repaired on an emergency basis via subcoronal incision and absorbable suture with simultaneous repair of eventual urethral lesion. Patients' status before fracture and voiding and erectile functions at long term were assessed by periodic follow-up and phone call. Detailed history included cause, symptoms, and single-question self-report of erectile and voiding functions. Results. Among the 44 suspicious cases, 42 (95.4%) were confirmed, mean age was 34.5 years (range: 18-60), mean follow-up 59.3 months (range 9-155). Half presented the classical triad of audible crack, detumescence, and pain. Heterosexual intercourse was the most common cause (28 patients, 66.7%), followed by penile manipulation (6 patients, 14.3%), and homosexual intercourse (4 patients, 9.5%). Woman on top was the most common heterosexual position (n = 14, 50%), followed by doggy style (n = 8, 28.6%). Four patients (9.5%) maintained the cause unclear. Six (14.3%) patients had urethral injury and two (4.8%) had erectile dysfunction, treated by penile prosthesis and PDE-5i. No patient showed urethral fistula, voiding deterioration, penile nodule/curve or pain. Conclusions. Woman on top was the potentially riskiest sexual position (50%). Immediate surgical treatment warrants long-term very low morbidity.

Up-regulation Of Dnam-1 And Nkp30, Associated With Improvement Of Nk Cells Activation After Long-term Culture Of Mononuclear Cells From Patients With Ovarian Neoplasia.

This study aimed at evaluating the functional activation and activating receptors expression on resting, short- and long-term NK and NK-like T cells from blood of ovarian neoplasia patients. Blood from patients with adnexal benign alterations (n = 10) and ovarian cancer (grade I-IV n = 14) were collected after signed consent. Effector cells activation was evaluated by the expression of the CD107a molecule. Short-term culture was conducted overnight with IL-2 and long-term culture for 21 days, by a method designed to expand CD56(+) lymphocytes. Short-term culture significantly increased NK cells activation compared to resting NK cells (p<0.05), however, the long-term procedure supported an even higher increase (p<0.001). Resting NK-like T cells showed poor activation, which was not altered by the culture procedures. The long-term culture effectively increased the expression of the activating receptors on NK and NK-like T cells, either by increasing the number of cells expressing a given receptor and/or by up-regulating their expression intensity. As a conclusion, the long-term culture system employed, resulted in a high number of functional NK cells. The culture system was particularly efficient on the up-regulation of NKp30 and DNAM-1 receptors on NK cells.

Genetic Predictors Of Long-term Response To Growth Hormone (gh) Therapy In Children With Gh Deficiency And Turner Syndrome: The Influence Of A Socs2 Polymorphism.

There is great interindividual variability in the response to GH therapy. Ascertaining genetic factors can improve the accuracy of growth response predictions. Suppressor of cytokine signaling (SOCS)-2 is an intracellular negative regulator of GH receptor (GHR) signaling. The objective of the study was to assess the influence of a SOCS2 polymorphism (rs3782415) and its interactive effect with GHR exon 3 and -202 A/C IGFBP3 (rs2854744) polymorphisms on adult height of patients treated with recombinant human GH (rhGH). Genotypes were correlated with adult height data of 65 Turner syndrome (TS) and 47 GH deficiency (GHD) patients treated with rhGH, by multiple linear regressions. Generalized multifactor dimensionality reduction was used to evaluate gene-gene interactions. Baseline clinical data were indistinguishable among patients with different genotypes. Adult height SD scores of patients with at least one SOCS2 single-nucleotide polymorphism rs3782415-C were 0.7 higher than those homozygous for the T allele (P < .001). SOCS2 (P = .003), GHR-exon 3 (P= .016) and -202 A/C IGFBP3 (P = .013) polymorphisms, together with clinical factors accounted for 58% of the variability in adult height and 82% of the total height SD score gain. Patients harboring any two negative genotypes in these three different loci (homozygosity for SOCS2 T allele; the GHR exon 3 full-length allele and/or the -202C-IGFBP3 allele) were more likely to achieve an adult height at the lower quartile (odds ratio of 13.3; 95% confidence interval of 3.2-54.2, P = .0001). The SOCS2 polymorphism (rs3782415) has an influence on the adult height of children with TS and GHD after long-term rhGH therapy. Polymorphisms located in GHR, IGFBP3, and SOCS2 loci have an influence on the growth outcomes of TS and GHD patients treated with rhGH. The use of these genetic markers could identify among rhGH-treated patients those who are genetically predisposed to have less favorable outcomes.

Long-term Postoperative Atrophy Of Contralateral Hippocampus And Cognitive Function In Unilateral Refractory Mtle With Unilateral Hippocampal Sclerosis.

This study aimed to evaluate long-term atrophy in contralateral hippocampal volume after surgery for unilateral MTLE, as well as the cognitive outcome for patients submitted to either selective transsylvian amygdalohippocampectomy (SelAH) or anterior temporal lobe resection (ATL). We performed a longitudinal study of 47 patients with MRI signs of unilateral hippocampal sclerosis (23 patients with right-sided hippocampal sclerosis) who underwent surgical treatment for MTLE. They underwent preoperative/postoperative high-resolution MRI as well as neuropsychological assessment for memory and estimated IQ. To investigate possible changes in the contralateral hippocampus of patients, we included 28 controls who underwent two MRIs at long-term intervals. The volumetry using preoperative MRI showed significant hippocampal atrophy ipsilateral to the side of surgery when compared with controls (p<0.0001) but no differences in contralateral hippocampal volumes. The mean postoperative follow-up was 8.7 years (± 2.5 SD; median=8.0). Our patients were classified as Engel I (80%), Engel II (18.2%), and Engel III (1.8%). We observed a small but significant reduction in the contralateral hippocampus of patients but no volume changes in controls. Most of the patients presented small declines in both estimated IQ and memory, which were more pronounced in patients with left TLE and in those with persistent seizures. Different surgical approaches did not impose differences in seizure control or in cognitive outcome. We observed small declines in cognitive scores with most of these patients, which were worse in patients with left-sided resection and in those who continued to suffer from postoperative seizures. We also demonstrated that manual volumetry can reveal a reduction in volume in the contralateral hippocampus, although this change was mild and could not be detected by visual analysis. These new findings suggest that dynamic processes continue to act after the removal of the hippocampus, and further studies with larger groups may help in understanding the underlying mechanisms.

Reasons For Brazilian Women To Switch From Different Contraceptives To Long-acting Reversible Contraceptives.

Long-acting reversible contraceptives (LARCs) include the copper-releasing intrauterine device (IUD), the levonorgestrel-releasing intrauterine system (LNG-IUS) and implants. Despite the high contraceptive efficacy of LARCs, their prevalence of use remains low in many countries. The objective of this study was to assess the main reasons for switching from contraceptive methods requiring daily or monthly compliance to LARC methods within a Brazilian cohort. Women of 18-50 years of age using different contraceptives and wishing to switch to a LARC method answered a questionnaire regarding their motivations for switching from their current contraceptive. Continuation rates were evaluated 1 year after method initiation. Sample size was calculated at 1040 women. Clinical performance was evaluated by life table analysis. The cutoff date for analysis was May 23, 2013. Overall, 1167 women were interviewed; however, after 1 year of use, the medical records of only 1154 women were available for review. The main personal reason for switching, as reported by the women, was fear of becoming pregnant while the main medical reasons were nausea and vomiting and unscheduled bleeding. No pregnancies occurred during LARC use, and the main reasons for discontinuation were expulsion (in the case of the IUD and LNG-IUS) and a decision to undergo surgical sterilization (in the case of the etonogestrel-releasing implant). Continuation rate was ~95.0/100 women/year for the three methods. Most women chose a LARC method for its safety and for practical reasons, and after 1 year of use, most women continued with the method.