Many cancerous solid tumors metastasize to the bone and induce pain (cancer-induced bone pain [CIBP]). Cancer-induced bone pain is often severe because of enhanced inflammation, rapid bone degradation, and disease progression. Opioids are prescribed to manage this pain, but they may enhance bone loss and increase tumor proliferation, further compromising patient quality of life. Angiotensin-(1-7) (Ang-(1-7)) binds and activates the Mas receptor (MasR). Angiotensin-(1-7)/MasR activation modulates inflammatory signaling after acute tissue insult, yet no studies have investigated whether Ang-(1-7)/MasR play a role in CIBP. We hypothesized that Ang-(1-7) inhibits CIBP by targeting MasR in a murine model of breast CIBP. 66.1 breast cancer cells were implanted into the femur of BALB/cAnNHsd mice as a model of CIBP. Spontaneous and evoked pain behaviors were assessed before and after acute and chronic administration of Ang-(1-7). Tissues were collected from animals for ex vivo analyses of MasR expression, tumor burden, and bone integrity. Cancer inoculation increased spontaneous pain behaviors by day 7 that were significantly reduced after a single injection of Ang-(1-7) and after sustained administration. Preadministration of A-779 a selective MasR antagonist prevented this reduction, whereas pretreatment with the AT(2) antagonist had no effect; an AT(1) antagonist enhanced the antinociceptive activity of Ang-(1-7) in CIBP. Repeated Ang-(1-7) administration did not significantly change tumor burden or bone remodeling. Data here suggest that Ang-(1-7)/MasR activation significantly attenuates CIBP, while lacking many side effects seen with opioids. Thus, Ang-(1-7) may be an alternative therapeutic strategy for the nearly 90% of patients with advanced-stage cancer who experience excruciating pain.

Many haematological malignancies are associated with widespread alterations of the transcriptional and epigenetic programmes. Changes in DNA methylation provide the clearest example of epigenetic changes, but the mechanism(s) underlying such changes is unknown. To investigate this I studied DNA methylation across an ~80kb segment of the genome which is not known to be mutated in haematological malignancies. Methylation was perturbed in 35-100% of samples of DNA from individuals with a wide range of haematological malignancies but not in non-malignant haematological disorders. DNA methylation was comprehensively assessed by Southern blot analysis, classical bisulphite sequencing and using a newly developed capture bisulphite sequencing protocol. The results were also compared with analysis by MeDIP, an immunoprecipitation-based technique. These analyses provide methylation status at various levels including individual CpG resolution. This showed both gain and loss of methylation at CpG dinucleotides. Of interest, hypomethylation was most frequently seen in intergenic regions corresponding to transcription factor binding sites and areas of increased chromosome accessibility. These observations suggested that hypomethylation of the genome in haematological malignancies could arise from aberrantly expressed DNA binding proteins which, recruited to sequences in regions of open chromatin, would protect the underlying CpG dinucleotides from the methylation machinery. This, in turn, could lead to passive demethylation accumulating with increasing cell divisions. This hypothesis was tested with electrophoretic mobility shift assays using oligonucleotides representing the DNA underlying one such region. This showed that, compared to nuclear extracts from the lymphocytes of normal individuals, those from patients with CLL were enriched for a protein which binds to oligonucleotides containing the underlying sequence. Using a mass spectrometry approach, I identified a variety of proteins that may bind such regions and account for their passive demethylation in haematological malignancies.

Background: Postural Orthostatic Tachycardia Syndrome (POTS) is a disorder of the autonomic nervous system that has high association with chronic pain syndromes such as fibromyalgia, migraine disorders, and chronic abdominal pain in adolescents with the diagnosis. Many of these disorders are characterized as central sensitization disorders, or pathological pain memory mediated by neural plasticity. Ehlers Danlos Syndrome Type 3 (EDS-3), also called joint hypermobility syndrome (JHS) is a genetic disorder of the connective tissue that causes joint laxity and is also highly associated with chronic pain syndromes as well as POTS. Methods: This study proposed to characterize POTS as a disorder of central sensitization. The hypothesis, presented within the proposed theoretical model, demonstrates that JHS leads to chronic pain that results in central sensitization and autonomic nervous system dysfunction (POTS). Other factors that were evaluated were anxiety and function. A sample size of 40 adolescents between the ages of 12 and 19 years were recruited from the cardiology and pain clinics at Children’s National Medical Center. Analysis of data utilizing Wilcoxon, Chi square, Pearson correlation, and logistic regression tests were completed using SAS 9.3. Results: In comparison to those without POTS, there were no significant associations found between having the diagnosis of POTS and any other variable studied in the model. JHS had a stronger correlation with anxiety, central sensitization, both subjectively, and objectively with hyperalgesia on Aδ sensory nerve fiber when compared to those without JHS. Subjective central sensitization was highly correlated with anxiety, function, age, and female gender. Function and central sensitization had a significant association even when removing anxiety as a covariate. Conclusions: These findings suggest that joint hypermobility may be a factor that contributes to the development of central sensitization in individuals with chronic pain. Dysautonomia is likely not a disorder of central sensitization, but rather a variable related to joint hypermobility and chronic pain in ways yet to be discovered. As previously discussed in other literature, anxiety has strong associations with central sensitization and functional disability in chronic pain syndromes, and when treated effectively may increase function in those that suffer with these disorders.

BACKGROUND Inhalation therapy has proven to be the best way to control the asthma and chronic obstructive pulmonary disease symptoms. The most commonly used delivery system to control these symptoms is the metered-dose inhaler. The primary goal of this study is to demonstrate an association between incorrect inhaler use and patient age. METHODS This is a cross-sectional study, performed at Centro Médico Naval “Cirujano Mayor Santiago Távara”, in Callao, Peru, in 2014. Patients older than 18 years that used metered-dose inhalers were included. We used film recordings of patients using a metered-dose inhaler and compared their technique with the recommendations on the guidelines on the correct use of inhalers of the Spanish Society of Pneumology and Thoracic Surgery (SEPAR). The main variables measured were age and incorrect inhaler use. The results were analyzed with the Chi squared test for bivariate analysis, and for multivariate analysis we used the Poisson regression model with robust variance. RESULTS We included 378 patients in the analysis; 167 were older than 60 years. An association was found between incorrect inhalator technique and age (p=0.014) (PR 1.19 95% CI 1.03 to 1.37). The highest prevalence of incorrect technique was found in the young adult population (88%). There was no association between the incorrect technique and the person who taught it (p=0.114). Finally, this study showed that 81.2% of the study population presented an incorrect inhalation technique. CONCLUSIONS The percentage of incorrect inhaler use, in the general population is high. Even if we found no association between an incorrect technique and the person who taught it; still, there is a high percentage of errors and it was even demonstrated that being instructed by a pulmonologist does not guarantee a correct performance of metered-dose inhaler inhalations. / INTRODUCCIÓN La terapia inhalatoria ha demostrado ser la más rápida y eficaz para el control del asma y la enfermedad pulmonar obstructiva crónica. El inhalador de dosis medida es el más usado por la población. El objetivo de este estudio es evidenciar la asociación entre la técnica inhalatoria incorrecta y la edad. MÉTODOS Estudio observacional, analítico, de corte transversal realizado en Perú durante 2014. Se incluyeron pacientes desde los 18 años que utilizaran inhalador de dosis medida. Se utilizó una lista de verificación de pasos establecidos por la Sociedad Española de Neumología y Cirugía Torácica y filmaciones para evaluar la técnica inhalatoria de los pacientes. Las variables principales fueron la edad y la mala técnica inhalatoria práctica. Para el análisis bivariado se utilizó la prueba Chi cuadrado y para el análisis multivariado regresión de Poisson con varianza robusta. RESULTADOS Se incluyeron 378 pacientes; 167 fueron mayores de 60 años. El estudio reveló que el 81,2% de la población presentó una incorrecta técnica inhalatoria. Se encontró asociación entre la edad y la técnica inhalatoria incorrecta (p=0,014) (PR 1,19 con IC 95% 1,03-1,37). El grupo etario con mayor frecuencia de técnica incorrecta fue el de adultos jóvenes (88%). CONCLUSIONES La frecuencia de uso incorrecto del inhalador en la población es alta y esta característica predomina en el grupo de adultos jóvenes. A pesar de no haber asociación entre la persona que enseña la técnica inhalatoria y el desempeño de la misma, se demostró que existe alta frecuencia de errores, incluso en aquellos pacientes instruidos por un médico especialista.

PURPOSE: The purpose of this study is to describe the subgingival bacterial biodiversity in untreated chronic periodontitis patients through the use of next generation 16S rRNA molecular analysis, and to determine similarities or differences between deep and shallow pockets within the same patients. METHODS: The analysis involved paired subgingival plaque samples from 24 subjects diagnosed with Generalized Moderate to Severe Chronic Periodontitis. One sample was selected from a single site having a probing depth >5 mm (i.e. Deep Site), and the other from a site with a probing depth <3mm (i.e. Shallow Site) within each subject. Bacterial DNA amplification of the V4-V6 region of the 16S rRNA was performed. The amplicons were sequenced via 454 Roche Genome Sequencer FLX System. The identified sequences were evaluated, and then compared to calculated false discovery rates. RESULTS: A total of 119 independent microbial genera were identified within the samples analyzed. Seven genera were identified to be statistically significant (p<0.05) in their association to deep or shallow sites following t-test and boot strap randomization: Actinomyces (p=0.004), Methylobacterium (p=0.028), Veillonella (p=0.028), and Rothia (p=0.038), and Streptococcus (p=0.033) in Shallow sites; while Mycoplasma (p=0.007) and Fusobacterium (p=0.016) were associated with deep sites. However, taking into account the calculated false discovery rates, it is suggested that none of the 119 microbial genera identified in this study were significantly associated with either deep nor shallow sites. CONCLUSION: The microbial genera identified within this study to be associated with deep and shallow sites follows the traditional pattern anticipated from the literature. However, the calculated false discovery rates suggest that these results may have occurred by chance and not due to a true difference.

Background: Mindfulness-based cognitive therapy (MBCT) is a promising new alternative to anti-depressant medication (Kuyken et al., 2015) and whilst some qualitative studies have explored participants' experiences of MBCT, none yet have explored experiences of participants who are considering coming off their antidepressant medication alongside MBCT or how patients experience change in relationships with self, others and illness. Aim: To examine MBCT participants' experience of change across 24 months, particularly in relation to change in views of their self and their illness over time. Method: Thematic analysis of in-depth retrospective interviews with 42 participants, two years after attending an 8 week MBCT group with an invitation to taper their antidepressant medication (ADM). Each participant took part in one retrospective interview which was semi-structured and focused on experiences of MBCT and ADM over the previous two years since attending an MBCT group and how these have impacted on a change in self and experience of illness. Findings and conclusion: Four over-arching themes were found: taking control, relationships (with self, other and illness), rebuilding the self and shifts in perspective. The findings in the current study are very similar to those found in transformation in the physical chronic illness literature (e.g. Paterson et al., 1999). Perhaps MBCT could be the challenge which lead patients suffering from chronic depression towards change and creates a context in which patients can consider self and identity.

2. Abstract Adaptation to chronic hypoxia is in addition to ischemic preconditioning one of the two known cardioprotective mechanisms. The precise molecular basis of these processes is still not fully explained. There are some studies that suggest the possible involvement of the adenosinergic signaling system in this adaptation. In this work, we focused on the characterization of the adenosinergic system in the myocardium of rats adapted to two regimens of chronic hypoxia - a protective continuous normobaric hypoxia (CNH) and non-protective intermittent hypoxia (INH/R, 23 h hypoxia and 1 h normoxia). Initially, we compared the total amount of adenosine receptors in samples from different groups of adapted animals. We discovered changes mainly at A2B receptor, which increased at CNH and declined in INH/R. This result suggests the possible involvement of A2B receptors in cardioprotection afforded by adaptation to chronic hypoxia. Furthermore, we investigated the distribution of various types of adenosine receptors and transporters in the plasma membrane of cardiac cells. We observed that A2A and A3 localize in membrane microdomains together with membrane enzyme CD73 that produces adenosine in the extracellular space by degrading AMP. A1 and A2B receptors similarly as nucleoside transporters ENT1, ENT2 and...

Title: Analysis of Pharmacotherapy by patients with diagnosis of COPD Student: Marilena Kartali-Kaouni Tutor: Prof. RNDr. Jiri Vlcek, CSc Department of Social and Clinical Pharmacy, Charles University in Prague, Faculty of Pharmacy in Hradec Kralove Background: " Chronic Obstructive Pulmonary Disease (COPD) is characterized by airflow limitation that is not fully reversible. The airflow limitation is usually both progressive and associated with an abnormal inflammatory response of the lungs to noxious particles or gases". Tobacco smoking is the major risk factor in the development of COPD. COPD is a leading cause of morbidity and mortality worldwide. Aim: 1st from the current literature to understand the nature of COPD and obtain information about the aetiopathogenesis of the disease, diagnosis options and summarize the current view of strategies for achieving the goals of treatment. 2nd in a pilot study to analyze drug therapy in COPD patients visiting a pharmacy in Greece. Methods: 56 prescriptions with the diagnosis of COPD were collected during a period of 8 months from a Greek pharmacy. Information from the prescriptions with regard to COPD medications prescribed (active substances, trade names, strength, dosage scheme, pack size), patients characteristics (age and gender) and prescribing...

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Stela Pavlíčková Supervisor: PharmDr. Ludmila Melicharová Title of diploma thesis: Pain therapy with focus on pacients with oncological disease Pain therapy is a part of the oncological illnesses complex treatment. This therapy doesn't solve the cause of cancer, but significantly increase patient's quality of life during oncological treatment, even in the terminal stage. The main target of this work is to give basic information about pain treatment, focusing on the oncological pain, and elaborate a review of pharmacological groups and individual substances most used in oncological pain treatment. Basic pillar of the cancer pain treatment is pharmacotherapy, which goes out of the WHO three-grade analgesic ladder. Basic therapy is made of nonopioid analgesics. In case that they aren't enough, we can add weak opioids, which can be replaced by strong opioids afterwards. Analgesics are usually used repeatedly and often in combinations. Dosage interval depends on pharmacological properties, dosage form and patient condition. Well controlled pain phases are usually interrupted by very strong, cruel pain episodes, which are called break-through pain. Very strong pain, fully or partially...

Master of Science / Department of Statistics / Abigail Jager / A discrete-time Markov chain with stationary transition probabilities is often used for the purpose of investigating treatment programs and health care protocols for chronic disease. Suppose the patients of a certain chronic disease are observed over equally spaced time intervals. If we classify the chronic disease into n distinct health states, the movement through these health states over time then represents a patient’s disease history. We can use a discrete-time Markov chain to describe such movement using the transition probabilities between the health states. The purpose of this study was to investigate the case when the observation interval coincided with the cycle length of the Markov chain as well as the case when the observational interval and the cycle length did not coincide. In particular, we are interested in how the estimated transition matrix behaves as the ratio of observation interval and cycle length changes. Our results suggest that more estimation problems arose for small sample sizes as the length of observational interval increased, and that the deviation from the known transition probability matrix got larger as the length of observational interval increased. With increasing sample size, there were fewer estimation problems and the deviation from the known transition probability matrix was reduced.