Sternal wound dehiscence from intense coughing in a cardiac surgery patient: could it be prevented?

This case report is on a obese patient who underwent a coronary artery bypass. The immediate postoperative phase, on the 8th day, was characterized by sternal dehiscence with diastasis, from intense coughing. At present, for obese patients there are not any guidelines aimed to prevent the sternal diastasis. The use of a sternum support vest after median sternotomy could be a valuable tool in the postoperative care of our patients, in order to avoid chest wall hyperexpansion and to prevent sternal wound dehiscence and diastasis.

The development of a photoacoustic Instrument for the monitoring of oil and other hydrocarbons in water.

The present methods for the detection of oil in discharge water are based either on chemical analysis of intermittent samples or bypass pipelines with instrumentation to detect either dissolved or dispersed hydrocarbons by a variety of optical techniques including absorption, scattering and fluorescence. However, test have shown that no single instruments entirely meets either present needs or satisfies the requirements of the future more stringent legislation which may limit total hydrocarbon content to 30 ppm or even less. Hence, in this paper, a detector is devised which can detect both dissolved and dispersed oil products, has a high immunity to scattering and can operate in-line and harsh environments with a detection sensitivity of a few ppm throughout a wide range of operations.

Applied web traffic analysis for numerical encoding of SQL Injection attack features.

SQL Injection Attack (SQLIA) remains a technique used by a computer network intruder to pilfer an organisation’s confidential data. This is done by an intruder re-crafting web form’s input and query strings used in web requests with malicious intent to compromise the security of an organisation’s confidential data stored at the back-end database. The database is the most valuable data source, and thus, intruders are unrelenting in constantly evolving new techniques to bypass the signature’s solutions currently provided in Web Application Firewalls (WAF) to mitigate SQLIA. There is therefore a need for an automated scalable methodology in the pre-processing of SQLIA features fit for a supervised learning model. However, obtaining a ready-made scalable dataset that is feature engineered with numerical attributes dataset items to train Artificial Neural Network (ANN) and Machine Leaning (ML) models is a known issue in applying artificial intelligence to effectively address ever evolving novel SQLIA signatures. This proposed approach applies numerical attributes encoding ontology to encode features (both legitimate web requests and SQLIA) to numerical data items as to extract scalable dataset for input to a supervised learning model in moving towards a ML SQLIA detection and prevention model. In numerical attributes encoding of features, the proposed model explores a hybrid of static and dynamic pattern matching by implementing a Non-Deterministic Finite Automaton (NFA). This combined with proxy and SQL parser Application Programming Interface (API) to intercept and parse web requests in transition to the back-end database. In developing a solution to address SQLIA, this model allows processed web requests at the proxy deemed to contain injected query string to be excluded from reaching the target back-end database. This paper is intended for evaluating the performance metrics of a dataset obtained by numerical encoding of features ontology in Microsoft Azure Machine Learning (MAML) studio using Two-Class Support Vector Machines (TCSVM) binary classifier. This methodology then forms the subject of the empirical evaluation.

Novas estratégias para entender o sistema de influxo de antibióticos e a resistência antimicrobiana a nível molecular

As fluoroquinolonas são antibióticos que têm um largo espectro de ação contra bactérias, especialmente Gram-negativas. O seu mecanismo de ação assenta na inibição de enzimas responsáveis pela replicação do DNA. Porém, devido ao seu uso indevido, o surgimento de resistência bacteriana a estes antibióticos tem-se tornado um grave problema de saúde pública. Uma vez que os seus alvos de ação se situam no meio intracelular, a redução da permeabilidade da membrana externa de bactérias Gram-negativas constitui um dos mecanismos de resistência mais conhecidos. Esta redução é associada à baixa expressão ou mutações em porinas necessárias para permitir o seu transporte, mais concretamente, da OmpF. Estudos prévios demonstraram que a coordenação de fluoroquinolonas com iões metálicos divalentes e 1,10-fenantrolina (genericamente designados metaloantibióticos) são potenciais candidatos como alternativa às fluoroquinolonas convencionais. Estes metaloantibióticos exibem um efeito antimicrobiano comparável ou superior à fluoroquinolona na forma livre, mas parecem ter uma via de translocação diferente, independente de porinas. Estas diferenças no mecanismo de captura podem ser fundamentais para contornar a resistência bacteriana. De forma a compreender o papel dos lípidos no mecanismo de entrada dos metaloantibióticos, estudou-se a interação e localização dos metaloantibióticos da Ciprofloxacina (2ª geração), da Levofloxacina (3ª geração) e Moxifloxacina (4ª geração) com um modelo de membranas de Escherichia coli desprovido de porinas. Estes estudos foram realizados através de técnicas de espectroscopia de fluorescência, por medições em modo estacionário e resolvida no tempo. Os coeficientes de partição determinados demonstraram uma interação mais elevada dos metaloantibióticos relativamente às respetivas fluoroquinolonas na forma livre, um facto que está diretamente relacionado com as espécies existentes em solução a pH fisiológico. Os estudos de localização mostraram que estes metaloantibióticos devem estar inseridos na membrana bacteriana, confirmando a sua entrada independente de porinas. Este mecanismo de entrada, pela via hidrofóbica, é potenciado por interações eletrostáticas entre as espécies catiónicas de metaloantibiótico que existem a pH 7,4 e os grupos carregados negativamente dos fosfolípidos da membrana. Desta forma, os resultados obtidos neste estudo sugerem que a via de entrada dos metaloantibióticos e das respetivas fluoroquinolonas deve ser diferente. Os metaloantibióticos são candidatos adequados para a realização de mais testes laboratoriais e uma alternativa promissora para substituir as fluoroquinolonas convencionais, uma vez que parecem ultrapassar um dos principais mecanismos de resistência bacteriana a esta classe de antibióticos.

Bowel-associated dermatosis-arthritis syndrome in an adolescent with short bowel syndrome

Bowel-associated dermatosis-arthritis syndrome (BADAS) is a neutrophilic dermatosis, characterized by the occurrence of arthritis and skin lesions related to bowel disease with or without bowel bypass. We report an unusual case of BADAS in a 15-year-old white male with congenital aganglionosis of the colon and hypoganglionosis of the small intestine and multiple bowel surgeries in childhood complicated by short bowel syndrome. He presented with recurrent peripheral polyarthritis, tenosynovitis, and painful erythematous subcutaneous nodules located on the dorsolateral regions of the legs and on the dorsa of the feet. Histological examination disclosed a neutrophilic dermatosis confirming the diagnosis of BADAS.Although an uncommon disease, especially at pediatric age, it is important to evoke the diagnosis of BADAS in children and adolescents with bowel disease, because treatment options and prognosis are distinct from other rheumatologic conditions.

The metabolic and epigenetic effects of the isocitrate dehydrogenase-1 mutation in glioma

Cancer cells have been noted to have an altered metabolic phenotype for over ninety years. In the presence of oxygen, differentiated cells predominately utilise the tricarboxylic acid (TCA) cycle and oxidative phosphorylation to efficiently produce energy and the metabolites necessary for protein and lipid synthesis. However, in hypoxia, this process is altered and cells switch to a higher rate of glycolysis and lactate production to maintain their energy and metabolic needs. In cancer cells, glycolysis is maintained at a high rate, even in the presence of oxygen; a term described as “aerobic glycolysis”. Tumour cells are rapidly dividing and have a much greater need for anabolism compared to normal differentiated cells. Rapid glucose metabolism enables faster ATP production as well as a greater redistribution of carbons to nucleotide, protein, and fatty acid synthesis, thus maximising cell growth. Recently, other metabolic changes, driven by mutations in genes related to the TCA cycle, indicate an alternative role for metabolism in cancer, the “oncometabolite”. This is where a particular metabolite builds up within the cell and contributes to the tumorigenic process. One of these genes is isocitrate dehydrogenase (IDH) IDH is an enzyme that forms part of the tricarboxylic acid (TCA) cycle and converts isocitrate to α-ketoglutarate (α-KG). It exists in three isoforms; IDH1, IDH2 and IDH3 with the former present in the cytoplasm and the latter two in the mitochondria. Point mutations have been identified in the IDH1 and IDH2 genes in glioma which result in a gain of function by converting α-KG to 2-hydroxyglutarate (2HG), an oncometabolite. 2HG acts as a competitive inhibitor of the α-KG dependent dioxygenases, a superfamily of enzymes that are involved in numerous cellular processes such as DNA and histone demethylation. It was hypothesised that the IDH1 mutation would result in other metabolic changes in the cell other than 2HG production, and could potentially identify pathways which could be targeted for therapeutic treatment. In addition, 2HG can act as a potential competitive inhibitor of α-KG dependent dioxygenases, so it was hypothesised that there would be an effect on histone methylation. This may alter gene expression and provide a mechanism for tumourogenesis and potentially identify further therapeutic targets. Metabolic analysis of clinical tumour samples identified changes associated with the IDH1 mutation, which included a reduction in α-KG and an increase in GABA, in addition to the increase in 2HG. This was replicated in several cell models, where 13C labelled metabolomics was also used to identify a possible increase in metabolic flux from glutamate to GABA, as well as from α-KG to 2HG. This may provide a mechanism whereby the cell can bypass the IDH1 mutation as GABA can be metabolised to succinate in the mitochondria by GABA transaminase via the GABA shunt. JMJ histone demethylases are a subset of the α-KG dependent dioxygenases, and are involved in removing methyl groups from histone tails. Changes in histone methylation are associated with changes in gene expression depending on the site and extent of chemical modification. To identify whether the increase in 2HG and fall in α-KG was associated with inhibition of histone demethylases a histone methylation screen was used. The IDH1 mutation was associated with an increase in methylation of H3K4, which is associated with gene activation. ChiP and RNA sequencing identified an increase in H3K4me3 at the transcription start site of the GABRB3 subunit, resulting in an increase in gene expression. The GABRB3 subunit forms part of the GABA-A receptor, a chloride channel, which on activation can reduce cell proliferation. The IDH1 mutation was associated with an increase in GABA and GABRB3 subunit of the GABA-A receptor. This raises the possibility of GABA transaminase as a potential therapeutic target. Inhibition of this enzyme could reduce GABA metabolism, potentially reducing any beneficial effect of the GABA shunt in IDH1 mutant tumours, and increasing activation of the GABA-A receptor by increasing the concentration of GABA in the brain. This in turn may reduce cell proliferation, and could be achieved by using Vigabatrin, a GABA transaminase inhibitor licensed for use in epilepsy.

Gestação após gastroplastia para tratamento de obesidade mórbida: série de casos e revisão da literatura

relatar a evolução de uma série de casos de gestação em mulheres previamente submetidas à cirurgia de bypass gástrico para tratamento de obesidade grave. Métodos: cinco casos consecutivos de gravidez após gastroplastia ocorridos entre 2001 e 2004 foram avaliados. As pacientes tinham idade entre 30 e 34 anos e todas haviam sido submetidas à cirurgia de Capella. Aspectos clínicos, laboratoriais e do acompanhamento materno e fetal foram considerados, durante o período gestacional e após o parto. Foi realizada revisão da literatura internacional, por meio das bases de dados MEDLINE e Web of Science, utilizando os seguintes unitermos: gastroplasty, gastric bypass surgery, bariatric surgery e pregnancy. Resultados: todas as gestações observadas foram únicas e não ocorreram complicações obstétricas, durante o seguimento pré-natal e parto. Também não houve registro de recém-nascidos prematuros ou de baixo peso ao nascimento. Conclusão: nossos dados sugerem que a gravidez após gastroplastia é segura para a mãe e feto. Entretanto, em virtude do limitado volume de informação disponível sobre o tema, investigações adicionais são necessárias para estabelecer recomendações apropriadas com relação ao seguimento dessas gestações

Identification and characterisation of telomere regulatory and signalling pathways after induction of telomere dysfunction

Telomeres are DNA-protein complexes which cap the ends of eukaryotic linear chromosomes. In normal somatic cells telomeres shorten and become dysfunctional during ageing due to the DNA end replication problem. This leads to activation of signalling pathways that lead to cellular senescence and apoptosis. However, cancer cells typically bypass this barrier to immortalisation in order to proliferate indefinitely. Therefore enhancing our understanding of telomere dysfunction and pathways involved in regulation of the process is essential. However, the pathways involved are highly complex and involve interaction between a wide range of biological processes. Therefore understanding how telomerase dysfunction is regulated is a challenging task and requires a systems biology approach. In this study I have developed a novel methodology for visualisation and analysis of gene lists focusing on the network level rather than individual or small lists of genes. Application of this methodology to an expression data set and a gene methylation data set allowed me to enhance my understanding of the biology underlying a senescence inducing drug and the process of immortalisation respectively. I then used the methodology to compare the effect of genetic background on induction of telomere uncapping. Telomere uncapping was induced in HCT116 WT, p21-/- and p53-/- cells using a viral vector expressing a mutant variant of hTR, the telomerase RNA template. p21-/- cells showed enhanced sensitivity to telomere uncapping. Analysis of a candidate pathway, Mismatch Repair, revealed a role for the process in response to telomere uncapping and that induction of the pathway was p21 dependent. The methodology was then applied to analysis of the telomerase inhibitor GRN163L and synergistic effects of hypoglycaemia with this drug. HCT116 cells were resistant to GRN163L treatment. However, under hypoglycaemic conditions the dose required for ablation of telomerase activity was reduced significantly and telomere shortening was enhanced. Overall this new methodology has allowed our group and collaborators to identify new biology and improve our understanding of processes regulating telomere dysfunction.

Role of catestatin as such as slowly released by fibronectin-coated pharmacologically active microcarriers (Fn-Pam) in limiting hypoxicinduced cell death

International audience

Effet préventif de la milrinone inhalée chez les patients avec hypertension pulmonaire subissant une chirurgie cardiaque sous circulation extracorporelle : une approche pharmacométrique

La circulation extracorporelle (CEC) est une technique utilisée en chirurgie cardiaque effectuée des milliers de fois chaque jour à travers le monde. L’instabilité hémodynamique associée au sevrage de la CEC difficile constitue la principale cause de mortalité en chirurgie cardiaque et l’hypertension pulmonaire (HP) a été identifiée comme un des facteurs de risque les plus importants. Récemment, une hypothèse a été émise suggérant que l'administration prophylactique (avant la CEC) de la milrinone par inhalation puisse avoir un effet préventif et faciliter le sevrage de la CEC chez les patients atteints d’HP. Toutefois, cette indication et voie d'administration pour la milrinone n'ont pas encore été approuvées par les organismes réglementaires. Jusqu'à présent, la recherche clinique sur la milrinone inhalée s’est principalement concentrée sur l’efficacité hémodynamique et l'innocuité chez les patients cardiaques, bien qu’aucun biomarqueur n’ait encore été établi. La dose la plus appropriée pour l’administration par nébulisation n'a pas été déterminée, de même que la caractérisation des profils pharmacocinétiques (PK) et pharmacodynamiques (PD) suite à l'inhalation. L'objectif de notre recherche consistait à caractériser la relation exposition-réponse de la milrinone inhalée administrée chez les patients subissant une chirurgie cardiaque sous CEC. Une méthode analytique par chromatographie liquide à haute performance couplée à un détecteur ultraviolet (HPLC-UV) a été optimisée et validée pour le dosage de la milrinone plasmatique suite à l’inhalation et s’est avérée sensible et précise. La limite de quantification (LLOQ) était de 1.25 ng/ml avec des valeurs de précision intra- et inter-dosage moyennes (CV%) <8%. Des patients souffrant d’HP pour lesquels une chirurgie cardiaque sous CEC était prévue ont d’abord été recrutés pour une étude pilote (n=12) et, par la suite, pour une étude à plus grande échelle (n=28) où la milrinone (5 mg) était administrée par inhalation pré-CEC. Dans l'étude pilote, nous avons comparé l'exposition systémique de la milrinone peu après son administration avec un nébuliseur pneumatique ou un nébuliseur à tamis vibrant. L’efficacité des nébuliseurs en termes de dose émise et dose inhalée a également été déterminée in vitro. Dans l'étude à plus grande échelle conduite en utilisant exclusivement le nébuliseur à tamis vibrant, la dose inhalée in vivo a été estimée et le profil pharmacocinétique de la milrinone inhalée a été pleinement caractérisé aux niveaux plasmatique et urinaire. Le ratio de la pression artérielle moyenne sur la pression artérielle pulmonaire moyenne (PAm/PAPm) a été choisi comme biomarqueur PD. La relation exposition-réponse de la milrinone a été caractérisée pendant la période d'inhalation en étudiant la relation entre l'aire sous la courbe de l’effet (ASCE) et l’aire sous la courbe des concentrations plasmatiques (ASC) de chacun des patients. Enfin, le ratio PAm/PAPm a été exploré comme un prédicteur potentiel de sortie de CEC difficile dans un modèle de régression logistique. Les expériences in vitro ont démontré que les doses émises étaient similaires pour les nébuliseurs pneumatique (64%) et à tamis vibrant (68%). Cependant, la dose inhalée était 2-3 fois supérieure (46% vs 17%) avec le nébuliseur à tamis vibrant, et ce, en accord avec les concentrations plasmatiques. Chez les patients, en raison des variations au niveau des facteurs liés au circuit et au ventilateur causant une plus grande dose expirée, la dose inhalée a été estimée inférieure (30%) et cela a été confirmé après récupération de la dose de milrinone dans l'urine 24 h (26%). Les concentrations plasmatiques maximales (Cmax: 41-189 ng/ml) et l'ampleur de la réponse maximale ΔRmax-R0 (0-65%) ont été observées à la fin de l'inhalation (10-30 min). Les données obtenues suite aux analyses PK sont en accord avec les données publiées pour la milrinone intraveineuse. Après la période d'inhalation, les ASCE individuelles étaient directement reliées aux ASC (P=0.045). Enfin, notre biomarqueur PD ainsi que la durée de CEC ont été identifiés comme des prédicteurs significatifs de la sortie de CEC difficile. La comparaison des ASC et ASCE correspondantes a fourni des données préliminaires supportant une preuve de concept pour l'utilisation du ratio PAm/PAPm comme biomarqueur PD prometteur et justifie de futures études PK/PD. Nous avons pu démontrer que la variation du ratio PAm/PAPm en réponse à la milrinone inhalée contribue à la prévention de la sortie de CEC difficile.

Bidirectional cargo transport by microtubule-based molecular motors

We have reconstituted a simple in vitro system using only mammalian dynein and mammalian kinesin attached to a single cargo. These cargoes undergo saltatory motion typically seen in vivo, indicating that the motors engage in a tug-of-war. When the complex hits a barrier, the cargo often reverses direction. In some cases, it tries several up-and-back motions, during which time the dynein likely pulls the cargo onto a different protofilament, and is sometimes able to bypass the blockage. This explains why eliminating kinesin or dynein stops motion in both directions in vivo. We also find that mammalian dynein, but not kinesin, often takes backwards steps when under backward force. However, yeast dynein coupled with mammalian kinesin does not display these attributes, as expected.

Genetic Suppressors of mrp-5 Lethality in C. elegans

Heme is an essential cofactor in numerous proteins, but is also cytotoxic. Thus, directed pathways must exist for regulating heme homeostasis. C. elegans is a powerful genetic animal model for elucidating these pathways because it is a heme auxotroph. Worms acquire dietary heme though HRG-1-related importers, and intestinal export was demonstrated to be mediated by the ABC transporter MRP-5. Loss of mrp-5 results in embryonic lethality. Although heme transporters have been identified, there are significant gaps in our understanding for the heme trafficking beyond HRG-1 and MRP-5. To identify additional components, we conducted a forward genetic screen utilizing the null allele mrp-5(ok2067). Screening of 160,000 haploid genomes yielded thirty-two mrp-5(ok2067) suppressor mutants. Deep-sequencing variant analysis revealed three of the suppressors subunits of adapter protein complex 3 (AP-3). We now seek to identify mechanisms for how adaptor protein deficiencies bypass a defect in MRP-5-mediated heme export.

La déconstruction de l'onto-théologie par Jacques Derrida : détour littéraire et mise en relief de la matrice langagière comme "différance"

Le mouvement derridien de la différance marque la rupture avec l'affirmation de la métaphysique de la présence, avec l'autorité du signifié transcendantal. Dans cet univers mouvant de signifiants qui se renvoient perpétuellement les uns aux autres, la logique d'univocité se disloque. La "présence" n'est que fantomatique, s'esquissant au sein d'une chaîne ininterrompue de signifiants et se laissant toujours creuser par la marque d'un irréductible manque. Face au logocentrisme, corollaire de l'affirmation de la présence, l'écriture se veut siège et articulation de la trace, d'une origine qui ne peut être que raturée, véhicule d'une irrémédiable fêlure. La volet littéraire de la déconstruction a pour but de mettre en évidence le fonctionnement de l'"indécidabilité" du discours, soit une certaine ambivalence dans la signification qui caractérise tout texte. L'objectif principal de la présente recherche est de fournir une compréhension plus approfondie de la déconstruction en insistant sur l'ancrage langagier de tout texte. Le discours philosophique n'échappe ainsi pas au mécanisme différentiel du langage et de la dérive métaphorique. La parenté entre la perspective déconstructiviste derridienne et la conception mallarméenne du langage poétique semble frappante. La mise en oeuvre, par Mallarmé, d'une dislocation de l'espace textuel, son minutieux "creusement" du vers après renoncement à toute quête d'"Idéal", la mise en relief du leurre de l'appropriation langagière, voilà qui trouve un écho particulier dans les thèses derridiennes. La "mimésis" platonicienne se voit au travers du prisme de la "mimique" mallarméenne. La déconstruction poursuit son travail de "luxation" de l'oreille philosophique, insérant les philosophèmes dans la matrice langagière, les livrant ainsi au hasard du cheminement textuel et les confrontant à l'aporie. La philosophie n'a alors d'autre choix que d'abandonner ses prétentions transcendantales. La marche de la "différance" instaure une inexorable distance qui prive le sujet de tout rapport direct avec une origine assurée et lui ôte toute possibilité de maîtrise sur le monde. Au travers de la langue, se profile la question de l'altérité, de la relation dissymétrique qui nous lie à cet "autre", ce "tout-autre" qui nous fonde et nous constitue. L'accueil inconditionnel de cette altérité nous mènera à l'étude de la "religion", la déconstruction se tournant vers le "religieux" tout en effectuant un "retournement" habile de tout credo essentialiste.

Cirugía bariátrica en Hospital Nacional Rosales, seguimiento a corto plazo de una cohorte

La obesidad es una epidemia actual y la Organización Mundial de la Salud la ha considerado como enfermedad. Una herramienta para la pérdida de peso es la cirugía, conocida como bariátrica, que incluye métodos restrictivos, malabsortivos y mixtos. En el Hospital Nacional Rosales se realiza cirugía bariátrica desde el año 2005, por lo que los autores se plantean como objetivo evaluar los resultados hasta el año 2010. Materiales y métodos: se realizó un estudio descriptivo de seguimiento de una cohorte evaluando la evolución de los pacientes hasta su cita más larga desde la cirugía, hasta 5 años de evolución. Resultados: entre los años 2005 – 2010 se encontró un total de 33 pacientes operados de cirugía bariátrica, siendo el 81.8% del sexo femenino. Las técnicas utilizadas fueron: bypass con plicatura 63.6%, by pass clásico con 30.3 % y manga gástrica con 6.06%. La mediana de IMC preoperatorio fue de 50 kg/mt². 72.7% tenían síndrome metabólico. La media de la pérdida del exceso de peso final fue de 48.9%, con un mínimo de 13.8% y una máxima de 82.2%. En el 100% de los pacientes diabéticos hubo reducción en los valores de glicemia. La tasa de complicaciones tempranas fue de 27.3%. La mortalidad fue de 4.7% para el by pass con plicatura, 10% para el bypass con reservorio y 50% para la manga gástrica. Conclusión: los resultados se encuentran dentro de los parámetros de eficacia y la morbilidad reportada en la curva de aprendizaje.

LHC main dipole magnet circuits: sustaining near-nominal beam energies

Crossing the Franco-Swiss border, the Large Hadron Collider (LHC), designed to collide 7 TeV proton beams, is the world's largest and most powerful particle accelerator the operation of which was originally intended to commence in 2008. Unfortunately, due to an interconnect discontinuity in one of the main dipole circuit's 13 kA superconducting busbars, a catastrophic quench event occurred during initial magnet training, causing significant physical system damage. Furthermore, investigation into the cause found that such discontinuities were not only present in the circuit in question, but throughout the entire LHC. This prevented further magnet training and ultimately resulted in the maximum sustainable beam energy being limited to approximately half that of the design nominal, 3.5-4 TeV, for the first three years of operation (Run 1, 2009-2012) and a major consolidation campaign being scheduled for the first long shutdown (LS 1, 2012-2014). Throughout Run 1, a series of studies attempted to predict the amount of post-installation training quenches still required to qualify each circuit to nominal-energy current levels. With predictions in excess of 80 quenches (each having a recovery time of 8-12+ hours) just to achieve 6.5 TeV and close to 1000 quenches for 7 TeV, it was decided that for Run 2, all systems be at least qualified for 6.5 TeV operation. However, even with all interconnect discontinuities scheduled to be repaired during LS 1, numerous other concerns regarding circuit stability arose. In particular, observations of an erratic behaviour of magnet bypass diodes and the degradation of other potentially weak busbar sections, as well as observations of seemingly random millisecond spikes in beam losses, known as unidentified falling object (UFO) events, which, if persist at 6.5 TeV, may eventually deposit sufficient energy to quench adjacent magnets. In light of the above, the thesis hypothesis states that, even with the observed issues, the LHC main dipole circuits can safely support and sustain near-nominal proton beam energies of at least 6.5 TeV. Research into minimising the risk of magnet training led to the development and implementation of a new qualification method, capable of providing conclusive evidence that all aspects of all circuits, other than the magnets and their internal joints, can safely withstand a quench event at near-nominal current levels, allowing for magnet training to be carried out both systematically and without risk. This method has become known as the Copper Stabiliser Continuity Measurement (CSCM). Results were a success, with all circuits eventually being subject to a full current decay from 6.5 TeV equivalent current levels, with no measurable damage occurring. Research into UFO events led to the development of a numerical model capable of simulating typical UFO events, reproducing entire Run 1 measured event data sets and extrapolating to 6.5 TeV, predicting the likelihood of UFO-induced magnet quenches. Results provided interesting insights into the involved phenomena as well as confirming the possibility of UFO-induced magnet quenches. The model was also capable of predicting that such events, if left unaccounted for, are likely to be commonplace or not, resulting in significant long-term issues for 6.5+ TeV operation. Addressing the thesis hypothesis, the following written works detail the development and results of all CSCM qualification tests and subsequent magnet training as well as the development and simulation results of both 4 TeV and 6.5 TeV UFO event modelling. The thesis concludes, post-LS 1, with the LHC successfully sustaining 6.5 TeV proton beams, but with UFO events, as predicted, resulting in otherwise uninitiated magnet quenches and being at the forefront of system availability issues.