A dengue é uma doença infecciosa transmitida pela picada de mosquitos do gênero Aedes. O vírus da dengue (DENV), pertencente ao gênero Flavivirus, família Flaviviridae, é atualmente um importante problema de saúde pública em todo o mundo. São reconhecidos quatro sorotipos antigenicamente distintos (DENV-1, -2, -3 e -4). A infecção por qualquer um dos sorotipos cursa de forma assintomática ou com quadro clínico que varia desde uma febre indiferenciada e autolimitada, passando pela febre clássica da dengue (FD) até quadros graves de febre hemorrágica da dengue (DHF). O diagnóstico clínico é difícil de ser realizado principalmente na faixa etária pediátrica em que os sintomas são muito similares aos de outras infecções febris agudas, ficando a cargo do laboratório o diagnóstico confirmatório. Este estudo descritivo do tipo série de casos teve como objetivo analisar as características clínicas e laboratoriais da dengue em pacientes pediátricos. A população de estudo foi constituída por 110 crianças com idade média de 9,3 ± 3,7 anos recrutadas em uma unidade de saúde no município de Ribeirão Preto, São Paulo. Amostras de sangue, saliva e urina foram coletadas das crianças 1-14 dias após o início do quadro clínico. Os sinais e sintomas mais frequentes foram a febre (105/110, 95%), seguida de cefaleia (66/110, 60%), mialgia (49/110, 45%), vômitos (27/110, 25%) e exantema (16/110, 15%). A infecção pelo vírus da dengue foi confirmada laboratorialmente em 96 crianças, as quais apresentavam sinais e sintomas compatíveis com a classificação de caso suspeito de dengue de acordo com os critérios da Organização Mundial da Saúde. A análise clínica inicial falhou em classificar como caso suspeito de dengue 46% das crianças. A carga viral foi significativamente maior no soro quando comparado àquela observada em saliva e urina, mas ainda assim estas amostras podem ser utilizadas como alternativa para diagnóstico da doença. O sequenciamento nucleotídico da região codificadora da proteína NS5 mostrou a circulação de DENV-3, principalmente durante o ano de 2010, e de DENV-1 e DENV-2, predominantemente no ano de 2011 na população do estudo. Houve predomínio de infecções primárias com quadro clínico leve sem complicações. / Dengue is an infectious disease transmitted by the biting of mosquitoes of Aedes genus. Dengue virus (DENV), belonging to the Flavivirus genus, Flaviviridae family, is an important public health problem worldwide. Four antigenically distinct viruses are recognized (DENV-1, -2, -3, e -4). Infection with any of the virus serotypes causes a spectrum of clinical manifestations ranging from inapparent or mid viral syndrome to classic dengue fever (DF) and severe hemorrhagic disease (DHF). The clinical diagnosis of dengue is difficult, especially in children because the symptoms are very similar to those observed in other febrile illness; thus, the confirmatory diagnosis is carried out by laboratory tests. This descriptive study aimed to analyze the clinical and laboratory features of dengue in pediatric patients. The study population consisted of 110 children; mean age 9.3 ± 3.7 years enrolled in a health center in Ribeirao Preto, Sao Paulo. Samples of blood, saliva and urine were collected from children 1-14 days after the onset of symptoms. The most common signs and symptoms were fever (105/110, 95%), followed by headache (66/110, 60%), myalgia (49/110, 45%), vomiting (27/110, 25%) and rash (16/110, 15%). Dengue virus infection was confirmed by laboratory tests in 96 children whom presented signs and symptoms compatible with the suspected dengue case classification in accordance with the criteria of the World Health Organization. The initial clinical examination failed to classify as a suspected dengue case 46% of children. The viral load in the serum was significantly higher when compared to saliva and urine. Even though, saliva and urine might be used as alternative samples for the diagnosis of the disease. The nucleotide sequencing of a partial region of NS5 protein gene showed the circulation of DENV-3, especially during the year 2010, and DENV-1 and DENV-2, predominantly in the year 2011 in the study population. There was a predominance of primary instead of secondary infections, all of them with self-limiting dengue fever.

Orientador: Paulo Cesar Rodrigues Palma / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-04T14:44:57Z (GMT). No. of bitstreams: 1 Thiel_RosanedoRocioCordeiro_M.pdf: 4989770 bytes, checksum: b7324d9b676908854ae181a6a48becc0 (MD5) Previous issue date: 2004 / Resumo: A incontinência urinária acomete cerca de 20% das mulheres na idade reprodutiva e afeta negativamente a qualidade de vida e a sexualidade. Entre agosto de 2002 e janeiro de 2004, 30 pacientes portadoras de incontinência urinária (lU), responderam os questionários ICIQSF( lntemational Consultation on lncontinence Questionnarie-Short Form) e FSFI (Female Sexual Function lndex) antes e após tratamento. A idade mediana foi de 43 anos (31 a 51 anos), com tempo de início dos sintomas urinários variando de 12 a 53 meses. Participaram pacientes com: incontinência urinária de esforço (59%), bexiga hiperativa idiopática (15%) e incontinência mista (26%). A avaliação estatística utilizou testes de Wilcoxon para dados pareados. Os valores do p :S0,05 indicaram diferenças significativas da RS antes e após o tratamento da lU. O objetivo foi avaliar a resposta sexual e a qualidade de vida de pacientes com incontinência urinária antes e após tratamento. A amostra final contou com 27 pacientes. A média de escore do ICIQ-SF passou de 17 para 7 após tratamento (p< 0,001) e a do escore ponderado do FSFI não teve alteração em nenhum domínio. Pacientes com BH apresentaram os menores escores ponderados de RS em todos os domínios. As submetidas à cirurgia apresentaram melhora dos escores ponderados nos domínios: desejo (p=0,02), satisfação (p=0,05) e no escore total (p=0,02). Treze pacientes com escore zero no ICIQ apresentaram aumento da média dos escores ponderados do FSFI nos domínios desejo (p<O,OI), satisfação (p=0,05) e no escore total (p=O,OI). É recomendada a avaliação da resposta sexual em pacientes com Incontinência urinária, porque esta condição causa impacto negativo na Qol / Abstract: The prevalence of urinary incontinence is around 20% of healthy middle-aged women, lncontinence causes negative impact on quality of life and sexuality. From August 2002 to January 2004, 30 patients (mean age 43 years) with stress urinary incontinence (59%), overactive bladder (15%) and mixed incontinence (26%) answered the ICIQSF( International Consultation on lncontinence Questionnarie-Short Fonn) and FSFI (Female Sexual Function lndex) questionnaires before and after treatment. The follow up ranged ITom12 to 53 months. Mean ICIQ score was 17 and 7 before and after treatment respectivally (p<O,OOl).Overactive bladder showed the worst scores in all domains. The patientes underwent to surgery increase desire (p=0,02), satisfaction (p=0,05) and total score (p=0,02). Thirteen patients wich ICIQ was zero increased: desire (p<O,Ol), satisfaction (p=0,05) and total score (p=O,Ol).Urinary incontinence significantly affects the qualityof life significantly. Sexualityevaluationin incontinencepatientsis recommend / Mestrado / Pesquisa Experimental / Mestre em Cirurgia

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23410号 / 医博第4755号 / 新制||医||1052(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 村川 泰裕, 教授 長尾 美紀, 教授 柳田 素子 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Drug-facilitated sexual assaults are a public health and safety concern. Liquid chromatography paired with tandem mass spectrometry is theoretically capable of detecting the scope of drugs commonly encountered in these types of cases. An analytical method was developed for the quantitative analysis of 40 drugs designated by Academy Standards Board 121 “Standard for the Analytical Scope and Sensitivity for Forensic Toxicological Testing of Urine in Drug Facilitated Crime” (ASB 121). The targeted analytes spanned a range of drug classes including antidepressants, antihistamines, barbiturates, benzodiazepines, cannabinoids, stimulants, and opioids. The final method utilized supported liquid extraction, followed by liquid chromatography tandem mass spectrometry with electrospray ionization in simultaneous positive and negative mode. Multiple reaction monitoring allowed quantification of analytes along with stable isotope internal standards. Validation parameters assessed included linearity, bias, precision, limit of detection, lower limit of quantitation, interference, and ion suppression or enhancement. The utilized sample preparation method was able to extract 36 of the 40 target analytes and the developed analytical method was able to detect and quantify all analytes to the sensitivities required by ASB 121.

As forensic scientists, we are required to accurately test for certain substances. This may necessitate the use of presumptive tests such as the One Step Multi-Drug Screen Test Card with the Integrated iCup®/iCup®A.D . There are many circumstances where these tests are applicable, such as job-related drug testing, custody and parole cases. An immunoassay, or presumptive test, is designed to give the analyst, even a non-scientific analyst, a general idea of what substance(s) are present in the individuals system, so that he or she is able to more accurately confirm what substances, if any, the individual may have used or consumed. The goal of the validation study of the One Step Multi-Drug Screen Test Card with the Integrated iCup®/iCup®A.D was to determine the sensitivity of various THC containing compounds (delta-9-THC, 11-nor-9-carboxy-delta9-THC, and 11-hydroxy-delta-9-THC) as well as different solutions containing Synthetic Cannabinoids at various concentrations and stored at different temperatures. Each of the drugs were tested below, at and above the cut-off of the drug stated by the manufacturer. The cut-off of 11-nor-9-carboxy-delta9-THC, given by the manufacturer, was 50 ng/mL. For every trial that was conducted, the drug could be detected in the iCup® at this limit of detection of 50 ng/mL, except when the drug had been stored in the freezer for approximately two months prior to use. Delta-9-THC was given a cut-off of 15,000 ng/mL, which is a high concentration, especially when these assays are used in custodial cases and job-related drug tests, where living individuals are providing a fresh specimen. The concentrations of delta-9-THC and 11-hydroxy-delta-9-THC were higher than the cut-off for a positive result of 11-nor-9-carboxy-delta9-THC, but it was tested below the 15,000 ng/mL cut-off for delta-9-THC, established by the manufacturer. After these adjustments were made, both delta-9-THC and 11-hydroxy-delta-9-THC could be detected in a range between 1,500 ng/mL and 5,000 ng/mL. While 1,500 ng/mL is still high for a living specimen, it is substantially lower than 15,000 ng/mL. Analyzing the higher concentration of the synthetic cannabinoid working stock solution of 10,000 ng/mL, positive results were detected at 3,500 ng/mL and 5,000 ng/mL. There were eight cannabinoids, metabolites, and synthetic cannabinoids found in the working stock solution: (1) THC, (2) 11-Hyroxy-delta-9-THC, (3) 11-nor-9-Carboxy-delta-9-THC, (4) AB-FUBINACA, (5) AB-FUBINACA Met. 3, (6) AB-FUBINACA Met. 2a, (7) AB-PINACA-blood, and (8) AB-PINACA Pentatonic Acid metabolite. As the concentrations decreased, a positive result was not produced. Ultimately, the final conclusions of all the testing was that the One Step Multi-Drug Screen Test Card with the Integrated iCup®/iCup®A.D is not as sensitive when it comes to the synthetic cannabinoids, the primary compound present in marijuana (delta-9-THC), and the active metabolite of marijuana (11-hydroxy-delta-9-THC). In order to gain more accurate results using this presumptive test, the sensitivity of the iCup® for a detection of delta-9-THC and 11-hydroxy-delta-9-THC at a lower concentration should be done. By, doing this, an analyst can be more confident when deciding what confirmatory test to use based on what substances are present in a given sample.

Urine samples are routinely used in human and animal patients to diagnose health problems; often to investigate or monitor specific health-related problems that essentially may remain silent for extended periods. However, not much work has been performed on crocodilian urine for diagnostics. In general, crocodilian species lack a bladder as a separate storage organ (as found in mammals), possess metanephric kidneys (unable to concentrate urine) and have functional salt excreting glands. Collection of urine from the Nile crocodile (Crocodylus niloticus) is a simple and atraumatic procedure where a dog urinary catheter is used to collect relatively clean urine from the urinary chamber in the crocodile’s cloaca. Unfortunately, in-depth investigations of urine variables, and establishing baseline concentrations, have not been performed on Nile crocodile urine samples before. The specific focus areas of this research project were: (1) determination of urine and plasma biochemical concentrations by means of a standard veterinary clinical pathology profile and the establishment of the ratio between urine and blood biochemical parameters; (2) the validation of a gas chromatographic–mass spectrometric (GC/MS) method for the determination of steroid metabolite concentrations in urine; and (3) using this established analytical method to determine the presence (identify) and concentrations of steroid metabolites in the urine of individual crocodilians. / Urine and plasma samples collected at Izintaba Crocodile Farm during the period November 2005 to July 2006, from captive bred, healthy young Nile crocodiles, were analysed for standard biochemistry variables. The urine samples (n = 101) were analysed for sodium, potassium, chloride, urea, creatinine, calcium, magnesium, phosphate, uric acid, osmolality, and ammonium ion, while the plasma samples (n = 101) were screened for total protein, glucose, sodium, potassium, chloride, urea, creatinine, total calcium, ionised calcium, magnesium, phosphate, uric acid and osmolality. Means, medians and standard deviations were statistically determined, as well as urine to plasma (U/P) ratios for corresponding variables. The value of this project is the establishment of reference concentrations for Nile crocodile urine samples that may become useful for interpretation of laboratory results, in future. / The clinical validation of a GC/MS method for the analysis of urinary steroids in the Nile crocodile was achieved using urine samples from two-year-old Nile crocodiles. The main objective of this investigation was to develop, optimize and validate the laboratory analysis of urinary steroid metabolites. Steroid profiling was performed on individual and pooled Nile crocodile urine samples. Ascending concentrations of representative steroid standards: androsterone, etiocholanolone, dehydroepiandrosterone, 11-OH androsterone, pregnanediol, pregnanetriol, 11-deoxytetrahydrocortisol, tetrahydrocortisone, tetrahydrocortisol and tetra-hydrocorticosterone, were spiked into aliquots of the pooled urine samples, to obtain calibration samples ranging from 0.2 to 20 μg. Sample preparation and analysis methodology were based on a well-established, validated GC/MS method for determination of human urinary steroid metabolites. The validation of the GC/MS method for Nile crocodile urine was successfully completed, by determining lower limits of quantitation and limits of detection for each analyte, obtaining linearity up to the highest calibration level, correlations exceeding 0.90, and recoveries of 82% and more. / Steroid profiling was performed on urine samples collected from a number of mature crocodilian species, namely Nile crocodile, American alligator (Alligator mississippiensis), spectacled caiman (Caiman crocodilus) and dwarf crocodile (Osteolaemus tetraspis). Steroid metabolites were identified and were quantitated and reported per urinary creatinine. Qualitative reporting was conducted in cases where creatinine concentrations were not available. Results included identification and quantitation of the steroid metabolites: androsterone, etiocholanolone, 11-hydroxy androsterone, pregnanediol, pregnanetriol, and the tetrahydro- metabolites of cortisone (THE), cortisol (THF), and corticosterone (THB). In some urinary steroid profiles, several prominent peaks were observed which could not be identified. The study findings confirmed that crocodile urine could successfully be used, as it is commonly used in humans, to determine steroid metabolite profiles. A follow-up study to identify the unknown peaks by structure elucidation with more sophisticated equipment is recommended - this could lead to valuable information about liver metabolism of steroids in crocodilians. / An adrenocorticotrophic hormone (ACTH) stimulation test was conducted on 18 captive Nile crocodiles. The experimental animals were temporarily housed in separate enclosures at Le Croc Crocodile Farm for four weeks, to ensure controlled conditions and easy and frequent access to the animals. Twenty-seven urine samples were collected both pre- and post-ACTH or saline injections. Steroid profiling was performed on 24 of the 27 urine samples to assess the corticosterone and tetrahydrocorticosterone concentrations following the ACTH treatment. Quantitation relative to urine creatinine levels was recorded following analyses with a standardised liquid chromatography/mass spectrometry (LC/MS) method, reporting the concentrations in nmol steroid/μmol creatinine. Unfortunately, a significant increase in urinary corticosterone concentrations 6 h after the injection of Synacthen® (5 μg/kg) was not observed. A possible explanation for this could be that the 6 h period was too short for a significant increase in urinary glucocorticoid metabolite excretion in the Nile crocodile. / In conclusion, this is the first in-depth study that focused, specifically, on Nile crocodile urine for analyses as diagnostic tools and for indices of health. The screening of the urine samples, collected from healthy Nile crocodiles, for a large array of biochemical variables contributed significantly to the database of “normal” concentrations. The establishment of a validated urinary steroid profiling method may significantly contribute to future validation and implementation of innovative diagnostic methods to monitor the health status and endocrine systems of wild Nile crocodiles in Africa. / Thesis (PhD)--University of Pretoria, 2017. / The Norwegian Council for Higher Education’s Programme for Development, Research and Education (NUFU) / Royal Netherlands Embassy in South Africa / Crocodile Specialist Group / SAVF / Paraclinical Sciences / PhD / Unrestricted

The existing linear economy approach to nutrient management has clear shortcomings including high expenditures for nutrient extraction and production of fertilizer as well as additional costs for nutrient removal at downstream waste water treatment plants (WWTPs) to prevent the pollution of aquatic environments. In a circular nutrient economy, phosphorus (P) and nitrogen (N) are removed from waste streams and captured as valuable fertilizer products in order to more sustainably reuse the resources in closed-loops and simultaneously protect receiving aquatic environments from harmful P and N emissions. The overarching aim of this thesis is to understand strategic approaches for nutrient recovery from wastewater and advance membrane technologies for P and N reclamation. The studies i.) approach nutrient recovery on a system-level to recognize optimal waste streams to target for P and N separation, ii.) advance membrane-based processes for nutrient recovery, and iii.) examine the economic viability of the nutrient recovery techniques.The thesis presents a thermodynamic and energy analysis of nutrient recovery from various waste streams of fresh and hydrolyzed urine, greywater, domestic wastewater, and secondary treated wastewater effluent. The analysis revealed comparative advantages in theoretical energy intensities for P and N recovery from nutrient-dense waste streams, such as fresh and hydrolyzed urine, compared to other more dilute sources. The thesis quantifies efficiencies required by separation techniques for nutrient reclamation to be competitive with the energy requirements of the prevailing industrial fertilizer production methods, i.e., phosphate mining and nitrogen fixation by the Haber-Bosch process. The dissertation examines and advances the performance of membrane-based processes for separation and recovery of P and N from diverted human urine. Donnan dialysis (DD), an ion-exchange membrane-based process, can capture and enrich orthophosphate, HxPO4(3−x)−, from source-separated urine. This work demonstrates the transport of Cl− driver ions down a concentration gradient, across an ion-exchange membrane to set up an electrochemical potential gradient that drives the transport of target HxPO4(3−x)− in the opposite direction, enabling P capture. Importantly, H2PO4− is transported against an orthophosphate concentration gradient, which achieves uphill transport of P. The thesis also provides a framework to better understand the impact of different ions in the water matrix on P recovery potential and kinetics. The thesis presents a novel operation of membrane distillation (MD) — isothermal membrane distillation with acidic collector (IMD-AC) — to selectively recover volatile ammonia, NH3, from hydrolyzed urine. The innovative isothermal and acidic collector features, respectively, suppressed undesired water permeation and enhanced ammonia vapor flux relative to conventional membrane distillation (CMD). The elimination of water flux in IMD-AC resulted in ≈95% savings in vaporization energy consumption relative to CMD. Critically, IMD-AC achieved uphill transport of ammoniacal nitrogen, i.e., transport against a concentration gradient, demonstrating the promising potential of the technique for N recovery. The dissertation proposes an integrated bipolar membrane electrodialysis (BPM-ED), DD, and IMD-AC system to drive the separation and recovery of orthophosphate and ammoniacal nitrogen from human urine. This work elucidates the role of pH and nutrient speciation (i.e., H2PO4− versus HPO42− and NH4+ versus NH3) on the performance of DD and IMD-AC. In the proposed configuration, BPM-ED generates acids and bases in situ to strategically control the pH of urine streams to benefit DD and IMD-AC performances. Strategic pH modification can enhance orthophosphate transport and selectivity in DD as well as ammonia transport and recovery potential in IMD-AC. Importantly, the analysis quantifies comparable specific energy consumptions of the proposed integrated membrane-based process to the existing approaches to P and N management. This thesis presents a preliminary economic assessment of onsite nutrient recovery employing DD and IMD-AC for respective P and N recovery from diverted urine. The analysis reveals opportunities to utilize widely-available waste chemical streams and recovered thermal energy to improve the economic viability of nutrient recovery. The largest capital expenditures are urine diversion toilets and additional piping for source-separation. Preliminary analysis demonstrates that employing urine diversion in public sanitation rooms, as opposed to private bathrooms, can reduce these capital expenditures. Furthermore, realizing savings from avoided costs for downstream nutrient removal at centralized wastewater treatment plants in addition to fertilizer revenue can enhance the economic viability of the approach. Overall, this dissertation critically informs nutrient recovery approaches and advances membrane-based processes for P and N reclamation to facilitate a paradigm shift from an inefficient linear nutrient economy to a sustainable circular nutrient economy. The work reveals opportunities to minimize energy intensity for nutrient separation, advance the performance of membrane-based techniques for selective and energy-efficient nutrient recovery from urine, and enhance the cost-competitiveness of nutrient reclamation. The findings of this work support nutrient recovery efforts and provide important insights that can be applied to other separation and resource recovery endeavors.

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The use of human excreta as a fertilizer has been used to a very limited extent. Human excreta has been distributed throughout the environment as a result of faulty sanitation system design, improper use and/or the total absence of any sanitation system. Human excreta contain nutrients in the form of nitrogen (N), phosphorus (P) and potassium (K). Agricultural use of sludge may be an alternative to conventional fertilizers provided that the risk to public health is considered. The application of biosolids to agricultural fields however poses an environmental and health risk since sludge may contain toxic chemicals, pollutant organic compounds and pathogenic parasitic microorganisms of faecal origin. If urine and faeces were separated the risks could potentially be minimized and both of these fractions could possibly be disinfected and utilized. Dehydrated faeces (humanure), urine, soil, irrigation water and crop samples were analysed for the total coliform, faecal coliform, faecal Streptococci, Salmonella spp, Aspergillus spp and helminth eggs. Moisture content and pH of each sample were also determined. Survival studies of microorganisms in dehydrated faeces using different treatments (ash; NaOH and pasteurization) were performed. Human urine both sterile and non-sterile was stored at 15oC, 20oC and 30oC for 50 d to estimate the effect of different storage conditions on the survival of pathogens. Four agricultural plots were prepared for treatment, two for spinach and two for carrot crops. Dehydrated faeces contained total coliform, faecal coliform, Faecal Streptococci and Salmonella spp. Treatment with NaOH and ash reduced the number of pathogens in dehydrated faeces, but not to safe levels. The number of helminth eggs was higher in dehydrated faeces (humanure) than in the soil and crops. Few helminth eggs survived pasteurization at 60oC for 30 min, 70oC for 20 min and 90oC for 5 min. Pathogens survived after storage of human urine at 15oC and 20oC for 50 d. Most of the pathogens (total coliform, faecal coliform, Salmonella spp and Aspergillus spp) were reduced at 30oC. 62.8% of helminth eggs found in humanure were Ascaris eggs and 82% ofAscaris eggs found were viable. Application of humanure to soil resulted in the contamination of soil and crops. However the viability of helminth eggs in crops was very low. No bacteria survived pasteurization at 70oC for 20 min and 90oC for 5 min. Therefore heat treatment at 70oC for 20 min and 90oC for 5 min was the most effective disinfection method for the bacteria. The results showed that Ascaris spp, Toxocara spp, Trichuris spp, Enterobius vermicularis, Hymnologies diminutionsand Taenia spp were present in soil, carrots, and spinach samples when humanure was applied to soil as a fertilizer. Pathogenic and parasitic microorganisms present in humanure represented a high risk of infection. / Dissertation (MSc (Microbiology))--University of Pretoria, 2005. / Microbiology and Plant Pathology / unrestricted