We explore a new laboratory technique in the field of urinalysis promising a combination of speed and selectivity in support of urinary tract infection diagnosis. Laboratory experimentation demonstrates long range surface plasmon polaritons (LRSPP) waveguides as a useful biosensor to selectively detect gram negative bacteria or gram positive bacteria in human urine. The biosensor can detect bacteria at concentration of 105 CFU/ml, the internationally recommended threshold for diagnostic of urinary tract infection (UTI). Using a negative control solution at bacterial concentration 1000x higher than the targeted bacteria in urine with a weak concentration of constituents, the power ratio between the negative control signals to the target bacteria signal is measured to be 5.4. Thus we report a conclusive demonstration of the LRSPP waveguide biosensor selectivity to the gram of bacteria in human urine. In addition, the biosensor may prove useful as an alternative urinalysis test method to determine the urine specific gravity, to estimate proteinuria, and to detect biofilm formation on surfaces.

Orientador: Adriano Fregonesi / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-26T20:23:11Z (GMT). No. of bitstreams: 1 Borges_ClaudioFerreira_D.pdf: 1990683 bytes, checksum: 1a75e1725e18e65f74c600f93224b4bb (MD5) Previous issue date: 2015 / Resumo: Objetivo: Foi realizada uma pesquisa de âmbito nacional com urologistas brasileiros a fim de estudar seu comportamento frente à exposição à radiação e ao uso de equipamentos de proteção e monitoramento. Material e Métodos: Um questionário com 13 perguntas foi enviado por e-mail para urologistas brasileiros; as questões abordavam temas como características demográficas, a exposição à radiação, bem como a utilização de dispositivos de proteção e dosímetros. As razões pelas quais esses dispositivos não foram utilizados também foram investigadas. Resultados: Um total de 332 questionários foi preenchido completamente e analisado; a idade média dos entrevistados foi de 43,3 ± 10,8 anos. Aventais de chumbo e protetores de tireoide são utilizados em cada procedimento por 84,4% e 53,89% dos entrevistados, respectivamente. Óculos de proteção nunca são utilizados por 72,12% dos entrevistados. Urologistas mais velhos são mais propensos a não usar proteção adequada. Dos urologistas que responderam, 76,42% nunca usam dosímetros. Conclusão: Este estudo mostra uma utilização insatisfatória dos dispositivos de proteção contra radiação e dosímetros, revelando uma baixa adesão ao ALARA (as low as reasonably achievable) entre urologistas brasileiros / Abstract: Purpose: We performed a nationwide survey of Brazilian urologists to study behaviors toward radiation exposure and the use of protective and monitoring equipment. Material and Methods: A 13-question e-mail questionnaire was sent to Brazilian urologists; the questions addressed demographic characteristics, radiation exposure, and the utilization of shielding devices and dosimeters. The reasons why these devices were not used were also investigated. Results: A total of 332 completed questionnaires were analyzed; the median age of the respondents was 43.3 ± 10.8 years. Lead aprons and thyroid protection are utilized in every procedure by 84.4% and 53.89% of respondents, respectively. Protective eyeglasses are never used by 72.12% of the respondents. Older urologists were more likely not to use adequate protection. Of the urologists who responded, 76.42% never use dosimeters. Conclusion: This study shows an unsatisfactory utilization of radiation-shielding devices and dosimeters, revealing a low compliance to the ALARA (as low as reasonably achievable) principle among Brazilian urologists / Doutorado / Fisiopatologia Cirúrgica / Doutor em Ciências

Artificial urinary sphincters have been used for decades for treatment of urinary incontinence. A commonly used device, the AMS 800 consists of a urethral cuff, pump and an abdominal reservoir. Notable complications of this system include scrotal or labial hematomas, infection or erosion of the cuff and rarely migration of its components. Although there are few reported cases related to effects from pump migration, those documenting reservoir related complications are even rarer. We present a case of reservoir migration adjacent to the ascending colon causing ischemic changes mimicking colonic neoplasm. Our patient, a 66-year old male with medical history of adenocarcinoma of prostate status post radical prostatectomy, had been having abdominal pain for a month. A CT scan showed cecal and proximal ascending colonic irregular nodular thickening suggestive of colonic mass. It also revealed a low-density structure next to the ascending colon abutting into area of the mass. A follow up colonoscopy showed a fungating, ulcerated mass extending from cecum to ascending colon concerning for a malignancy of which biopsy was also done. The patient then underwent right open hemicolectomy. During surgery, a balloon reservoir was seen in the abdominal cavity with its adherence to the right colon but not eroding into it. The surgeon dissected the balloon, repositioned and re-peritonealized it before closing the abdomen. The colonoscopic and surgical pathology instead demonstrated findings of ischemic colitis with mucosal ulceration in cecum and ascending colon limited to the mucosa but no evidence of cancer. Retrospective chart review revealed history of artificial urinary sphincter implantation for urinary incontinence related to radical prostatectomy for adenocarcinoma eight years prior. With manufacturer suggested implant location of the reservoir in prevesical space, the possibility of migration needs to be accounted for. Although there are not many reports of artificial sphincter reservoir related complications, there are cases documenting inflatable penile prosthesis reservoir erosion into abdominal and pelvic structures. As the CT scan demonstrated reservoir indentation into the ascending colon, it likely led to chronic irritation of the adjacent colonic wall due to mass effect. It is hypothesized that constant pressure on colonic wall likely led to localized ischemia. This resulted in localized inflammation including submucosal edema, which can create a mass-like appearance when severe. This case emphasizes that, while preliminary radiographic imaging and even gross colonoscopy findings may be suggestive of a malignancy, it is imperative to await biopsy results to confirm the diagnosis of a malignant neoplasm. Our case report emphasizes the consideration of diagnoses other than colon cancer when faced with a colonic mass especially in the setting of implanted intra-abdominal foreign body to avoid unnecessary surgery and related complications.

Orientador: Priscylla Tatiana Chalfun Guimarães Okamoto / Resumo: A doença do trato urinário inferior de felinos (DTUIF) corresponde a uma série de afecções que podem acometer a bexiga e a uretra desses animais. A forma obstrutiva é a consequência mais prevalente e mais grave dentre outras DTUIF, podendo levar o animal a azotemia pós-renal, redução da taxa de filtração glomerular (TFG) e a lesão renal aguda (LRA), que caso não seja tratada de forma eficiente, pode evoluir para doença renal crônica (DRC), uma doença frequente na população geriátrica felina. A creatinina sérica (sCr) é o biomarcador de TFG mais utilizado na clínica veterinária, porém apresenta baixa sensibilidade e diversos fatores que podem afetar seus valores. A dimetilarginina simétrica (SDMA) é um biomarcador mais recente na medicina veterinária que vem apresentado maior precocidade na detecção da perda da função renal e menor interferência de fatores extrarrenais, porém poucos são os trabalhos que determinam valores de SDMA em lesão renal de gatos. Este trabalho visou avaliar os valores de SDMA e compará-los com a sCr, ureia, dados hemogasométricos, escore clínico e tempo de obstrução de gatos com DTUIF obstrutiva. Os animais foram alocados em dois grupos experimentais, sendo 17 animais do grupo obstruído (GO) e 13 animais sadios para grupo controle (GC). As amostras foram coletadas antes da desobstrução (M0) e durante o tratamento clínico, nos momentos 12, 24 e 48 horas (M12, M24 e M48). Resultados obtidos demonstraram que no M48 do GO, 50% dos gatos obstruídos apresent... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Feline lower urinary tract disease (FLUTD) is a series of conditions that can affect the bladder and urethra of these animals. The obstructive form is the most prevalent and most serious consequence among other FLUTD, which can lead to post-renal azotemia and reduced glomerular filtration rate (GFR) and acute kidney injury (AKI), which if not treated effectively, can progress to chronic kidney disease (CKD), a common disease in the feline geriatric population. Serum creatinine (sCr) is the biomarker of GFR most commonly used in veterinary pratice, but it has low sensitivity and there are several factors that may affect its values. Symmetrical dimethylarginine (SDMA) is a newer biomarker in veterinary medicine that has been shown to be more precocious in detecting loss of renal function and less interference of extrarenal factors, but there are few studies determining SDMA values in feline kidney injury. This work aimed to evaluate SDMA values and to correlate them with sCr, urea, hemogasometric data, clinical score and duration of obstruction of felines with obstructive DTUIF. The animals were allocated into two experimental groups, 17 animals from the obstructed group (GO) and 13 healthy animals for control group (GC). Samples were collected before clearance (M0) and during clinical treatment at 12, 24 and 48 hours (M12, M24 and M48). The results obtained were that in GO M48, 50% of obstructed cats had SDMA values above normal, while for sCr only 29.41% of cats were elevated... (Complete abstract click electronic access below) / Mestre

Pelvic organ prolapse is characterized by the failure of vaginal wall support and protrusion of the pelvic organs through the vaginal orifice. Exact etiology of pelvic organ prolapse is not completely understood. The surgical procedures for pelvic organ prolapse utilize various biomaterials for holding the organs in place. However, the biomaterials used for restoring these organs have a high rate of failure in a complicated anatomical and biomechanical environment. With the given issues at hand, animal models are the best answer for understanding the pathophysiology of prolapse, and determining the cause of failure of these surgical interventions. For this study, we are investigating sheep as an animal model for human pelvic organ prolapse. We compared the anatomy of the sheep pelvic floor with humans. We found that anatomical parameters are a good measure/biomarker for estimating structural and anatomical changes in the body of the animal. As the anatomical measurements are applied to human vaginal prolapse, we can apply the same principles in sheep and further explore the feasibility of using sheep as an animal model for prolapse. Additionally, we evaluated location dependent biomechanical properties of the sheep vaginal tract. We have characterized the structure-property relationship of sheep vaginal wall tissue in the top third and middle third regions. We found that in contrast to current published research, sheep vaginal tissues are anisotropic in nature. This anisotropic characteristic of the sheep vaginal wall tissue is a direct function of the microstructural arrangement of collagen, elastin, smooth muscle and other extracellular matrix components. We also developed decellularized scaffolds as potential biomaterials, which can be potentially utilized in prolapse surgeries. We developed three different types of vaginal tissue scaffolds using SDS, Triton X-100, and trypsin for reconstructive surgery applications. During the decellularization, all of the cellular components are removed, which leaves the acellular ECM behind. We analyzed the biomechanical properties and microstructural properties of these scaffolds and found that the SDS samples were better in all aspects of the preclinical evaluation. Future studies will aim at applying the anatomical and biomechanical techniques used in this study to prolapsed sheep vaginal wall tissues.

Bladder dysfunction is seen as a public health problem with a strongly increasing incidence in older men. Despite this, the man's experience of bladder dysfunction is a topic that has not been researched in the empirical field as much as women’s experiences. Not highlighting the man's experience leads to suffering and constitutes an obstacle to equal care. The aim of this study was to illustrate the man's experience of living with bladder dysfunction. This study is intended to illustrate experiences, therefore it was well suited to make an integrative compilation of qualitative research – inspired by meta-synthesis. Nine qualitative articles that described the man's experiences were analysed schematically and resulted in three themes and six sub-themes. Experiences that were noticed in the man were the impact of the environment in bladder dysfunction which included environmental and health care responses. Emotional impact in bladder dysfunction, which highlighted the experiences that arose in connection with bladder dysfunction. The life adaptations in bladder dysfunction was an experience which described the changes the man made to achieve a normality in everyday life as well as changes to hide the condition. The man used a range of strategies to maintain normality in life and to keep his condition a secret from those around him. This is seen because of the stigmatization that emerged through this study. Healthcare failed to care for the man with bladder dysfunction and was seen as partially responsible for the stigmatization the man experienced.

<p><em>Background:</em> UC is the most common urogenital cancer, comprising up to 2% of all naturally occurring neoplasia in dogs and can be challenging to diagnose. With early diagnosis, the disease can be controlled in most dogs with a good quality of life. MiRNAs are small non-coding RNAs that function by post-transcriptional regulation of gene expression. Their abundant presence and stability in the body make them promising tools for disease diagnosis. </p> <p><em>Hypothesis:</em> A microRNA (miRNA) signature can be used to differentiate canine urothelial carcinoma (UC) from other lower urinary tract diseases.</p> <p><em>Literature review:</em> There is an overlap of miRNA expression changes between normal physiologic processes, non-infectious and non-inflammatory conditions, infectious and/or inflammatory conditions, and neoplasia. Additionally, the mechanism of action of these overlapping miRNAs varies depending on the disease process. There is a lack of standardization of miRNA evaluation and consistency within a single evaluation method. Herein we evaluate three papers on miRNA expression in canine UC and compared the reported expression profile to human UC literature and identified experimentally validated targets of the dysregulated miRNA. </p> <p><em>Methods and results:</em> <strong>(Aim 1)</strong> Using reverse transcriptase quantitative PCR (RT-qPCR), we assessed the effects of sample handling on miRNA expression in formalin-fixed Paraffin-embedded (FFPE) tissue and urine sediment. We showed that the time of tissue fixation in formalin does not alter the detection of miRNA expression, but the inclusion of the muscularis layer altered the miRNA expression profile in bladder tissue. Additionally, miRNAs in urine sediment were proven to be stable despite the storage temperature for up to two weeks. <strong>(Aim 2)</strong> Using Next Generation Sequencing (NGS) with validation of findings via RT-qPCR, we evaluated differential miRNA expression in bladder tissue collected from normal canine urothelium and the invasive type of UC (iUC) to elucidate the dysregulated pathways. We found that twenty-eight miRNAs were differentially expressed (DE). The DE miRNAs were most often associated with gene silencing by miRNA, miRNAs in cancer, and miRNAs involved in DNA damage responses. Proteins involved include HRAS, KRAS, ARAF, RAF1, MAPK1, MAP2K1, MAPK3, FGFR3, EGFR, HBEGF, RASSF1, E2F2, E2F3, ERBB2, SRC, MMP1, and UP3KA. <strong>(Aim 3)</strong> Using RT-qPCR, expression of miR-214, miR-181a, miR-361, and miR-145 were evaluated. We failed to reject the null hypothesis that the relative gene expression in all groups was the same for any miRNA, nor did we find any multivariate summary that could effectively differentiate UC from inflammatory and non-neoplastic transitional cells. </p> <p><em>Conclusions:</em>   The findings within this thesis highlight the need for standardized methods for miRNA evaluation, support the use of stored samples for miRNA expression analysis, and show the importance of isolating the tissue of interest in FFPE. We defined the miRNome of iUC and investigated numerous protein pathways affected by dysregulation of differentially expressed miRNA in urothelial carcinoma. While we failed to reject our null hypothesis that the miRNA signature we evaluated could be utilized as a diagnostic tool for canine urothelial carcinoma, we showed the promise of miRNA as diagnostic tools and highlight several novel pathways that miRNA regulation affects in this disease. </p>

Section A provides an overview of the role of family caregivers of individuals with chronic illness, and describes key conceptualisations and theories posited in the caregiver literature. This is followed by an overview of research conducted with caregivers of children with chronic kidney disease (CKD), a summary of the limitations of this research, and suggestions for future research. Section B Despite a recognised need to monitor caregiver burden in caregivers of children with CKD, there is no measurement tool currently available to meet this aim. The present research documents the development of a measure of caregiver burden specific to family caregivers of children with CKD. Methods: Interviews were conducted with 16 caregivers of children with CKD and 10 healthcare professionals in order to generate measure items. A provisional version of the measure was developed and piloted with 18 caregivers of children with CKD and five healthcare professionals. Results: An initial pool of 97 items was generated from the content of interviews, which was reduced to 60 items following review for item redundancy. A piloting exercise provided preliminary evidence for the usability, readability, and relevance of measure items; adaptations further to piloting resulted in the 51-item ‘Paediatric Renal Caregiver Burden Scale’ (PR-CBS). Conclusions: It is hoped that the PR-CBS will serve to identify areas of need amongst caregivers of children with CKD, and in turn improve outcomes for this caregiver population and children with CKD. Section C is a critical appraisal of the conducted research study, and includes an overview of research abilities acquired during its completion, reflections on how the research may have been conducted differently, implications for future clinical practice, and ideas for future research.

Aim: To study the possibility of detecting lymph node metastasis in locally advanced urinary bladder cancer (UBC) treated with radical cystectomy (RC) by using preoperative positron emission tomography/computed tomography (PET/CT) and peroperative sentinel node biopsy (SNB) technique. We also investigate the clinical significance of macrophage traits expression by cancer cells, M2-macrophage infiltration (MI) in tumor stroma and the immunohistochemical expression of biomarkers in cancer cells in relation to clinicopathologic data. Patients and Methods: We studied prospectively 122 patients with UBC, pathological stage pT1–pT4 treated with RC and pelvic lymph node dissection (PLND) during 2005–2011 at the Department of Urology, Linköping University Hospital. In the first study, we compared the results of preoperative PET/CT and conventional CT with the findings of postoperative histopathological evaluation of lymph nodes (LNs). In the second study we investigated the value of SNB technique for detecting pathological LNs during RC in patients with UBC. W also examined the significance of the primary tumor location in the bladder in predicting the site of LN metastases, and the prognostic significance of lympho-vascular invasion (LVI) and lymph node metastasis density (LNMD) on survival. In the third study, we investigate the clinical significance of macrophage infiltration (MI) in tumor stroma and macrophage-traits expression by tumor cells. In the fourth study, we investigate the cell cycle suppression proteins p53, p21, pRb, p16, p14 ARF as well as tumors proliferative protein Ki67 and DNA repair protein ERCC1 expression in cancer cells. The results were compared with clinical and pathological characteristics and outcome. Results: Prior to RC, PET/CT was used to detect LN metastasis in 54 patients. PET/CT had 41% sensitivity, 86% specificity, 58% PPV, and 76% NPV, whereas the corresponding figures for conventional CT were 41%, 89%, 64%, and 77%. SNB was performed during RC in 103 patients. A median number of 29 (range 7–68) nodes per patient were examined. SNs were detected in 83 out of 103 patients (81%). The sensitivity and specificity for detecting metastatic disease by SNB varied among LN stations, with average values of 67% -90%. LNMD or ≥8% and LVI were significantly related to shorter survival. In 103 patients, MI was high in 33% of cases, while moderate and low infiltration occurred in 42% and 25% of tumors respectively. Patients with tumors containing high and moderate compared to low MI had low rate of LN metastases (P=0.06) and improved survival (P=0.06), although not at significant level. The expression of different tumor suppression proteins was altered in 47-91% of the patients. There were no significant association between cancer specific survival (CSS) and any of the studied biomarkers. In case of altered p14ARF, ERCC1 or p21, CSS was low in case of low p53 immunostaining but increased in case of p53 accumulation, although not at a significant level, indicating a possible protective effect of p53 accumulation in these cases. Conclusion: PET/ CT provided no improvement over conventional CT in detection and localization of regional LN metastases in bladder cancer. It is possible to detect the SN but the technique is not a reliable for perioperative localization of LN metastases; however, LVI and LNMD at a cut-off level of 8% had significant prognostic values. MI in the tumor microenvironment but not CD163 expression in tumor cells seems to be synergistic with the immune response against urinary bladder cancer. Our results further indicate that altered p53 might have protective effect on survival in case of altered p14ARF, p21, or ERCC1 indicating an interaction between these biomarkers.

Rectal cancer is one of the three most common malignancies in Sweden with an annual incidence of about 2000 cases. Current treatment consists of surgical resection of the rectum including the loco-regional lymph nodes in the mesorectum. In advanced cases, neoadjuvant chemo-radiotherapy (CRT) prior to the operative treatment reduces local recurrences and enables surgery. The neoadjuvant treatment can also eradicate the tumour completely, i.e. complete response. This research project was designed to investigate the effects of preoperative radiotherapy/ CRT and analyze methods to predict response to CRT. Study I investigated the expression of the FXYD-3 protein with immunohistochemistry in rectal cancer, with or without preoperative radiotherapy. The results from the total cohort showed that, strong FXYD-3 expression was correlated to infiltrative tumour growth (p = 0.02). In the radiotherapy group, strong FXYD-3 expression was related to an unfavourable prognosis (p = 0.02). Tumours with strong FXYD-3 expression had less tumour necrosis (p = 0.02) after radiotherapy. FXYD-3 expression in the primary tumour was increased compared to normal mucosa (p=0.008). We concluded that FXYD-3 expression was a prognostic factor in patients receiving preoperative radiotherapy for rectal cancer. Study II investigated FXYD-3 expression in tumours that developed local recurrences following surgery and compared this with expression in tumours that did not develop local recurrences. There was no difference in the expression of FXYD-3 between the group that developed local recurrences and the group that did not develop local recurrences. There was no difference in survival between those with strong or weak FXYD-3 expression. We concluded that this study could not confirm the findings from study 1 i.e. that FXYD-3 expression has prognostic significance in rectal cancer. Study III was a register-based study on the incidence and effects of complete response to neoadjuvant treatment. Eight per cent of the patients with adequate CRT to achieve complete response also had a complete histological response of the luminal tumor in the resected bowel. Sixteen per cent of that group had remaining lymph node metastases in the operative specimen. Chemotherapy together with radiotherapy doubled the chance of complete response in the luminal tumour. Patients with remaining lymph node metastases had a lower survival rate compared to those without. We concluded that residual nodal involvement after neoadjuvant treatment was an important factor for reduced survival after complete response in the luminal tumour. Study IV followed up the results from the previous study by re-evaluating magnetic resonance imaging (MRI)- images in patients with complete tumour response. Two experienced MRI radiologists performed blinded re-staging of post CRT MR- images from patients with complete response in the luminal tumour. One group with lymph node metastases and another one without were studied and the results compared with the pathology reports. The sensitivity, specificity, and positive and negative predicted values for correct staging of positive lymph nodes was 37%, 84%, 70% and 57%. The size of the largest lymph node (4.5 mm, p=0.04) seemed to indicate presence of a tumour positive lymph node. We concluded that MRI couldn’t correctly stage patients for lymph node metastases in patients with complete response to CRT in the luminal tumour.