Alpha-1-microglobulin is co-synthesized with bikunin on the AMBP gene (

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Ammonium chloride (NH4Cl) has been the primary preventive modality for struvite urolithiasis in goats. This study investigated the effect of continuous and pulse dose NH4Cl therapeutic regimens on urine pH in ten goats. The initial regimen (feed additive) consisted of 0.007% NH4Cl as a feed additive. Following this week long regime, the two treatment regimens were designed as a standard ten goat cross-over design. The first treatment regimen (continuous) consisted of daily administration of a titrated dosage of NH4Cl for ten days, followed by four days without treatment. The third treatment regimen (pulse) used daily administration of a titrated dose of NH4Cl for three consecutive days followed by four days without treatment for three treatment periods. Ammonium chloride dosages were titrated to result in a urine pH of < 6.5 (target level) prior to commencing treatment regimens. Urine pH was evaluated once daily during feed additive regimen and twice daily during the treatment regimens. A Bayesian methodology was used to determine the daily odds ratios for production of target urine pH during treatment regimens. The odds ratios were also calculated between pulse dosages during the pulse regimen. The feed additive regimen did not result in target urine pH within 7 days. Treatment with the continuous regimen resulted in target pH, however, pH returned to >6.5 within five days, (odds ratio 0.23-1.56 at Treatment Time 10). The odds ratios for each pulse period of the pulse dose regimen were 2.20-7.45, 0.41-1.68 and 1.59-5.62 respectively. The results of this study indicate that variability in response to therapy warrants titrating individual dosages of NH4Cl, continuous therapy results in a loss of effectiveness, and pulse dosage is effective in repeatedly producing a urine pH of <6.5.

Urinary bladder marsupialization has been successful in producing acceptable long-term resolution of clinical cases of obstructive urolithiasis in male goats. The purpose of this study was to evaluate the six-month outcome of urinary bladder marsupialization in male goats. The urinary bladders of six male goats free from systemic disease were marsupialized following induced urethral obstruction. Renal ultrasonography, complete blood count, and blood chemistry analysis were evaluated preoperatively (day 0), at 7 postoperative days, and at 30-day intervals until 180 postoperative days. Stomal diameter was recorded at each interval. Necropsy examination was performed on day 180 or when stomal stricture or death occurred. Stomal stricture occurred in one goat at 120 days, and another goat was found dead at 150 days. Necropsy of this goat revealed severe, suppurative cystitis. All goats developed mild urine scald dermatitis. All blood chemistry values remained within normal limits. Significant decreases in white blood cell count, serum creatinine, and stomal diameter were observed from day 0 to day 180. Except for the goat that died at 150 days, all urinary bladders were tubular in shape and the mucosa and serosa of all urinary tract organs appeared grossly normal at necropsy examination. Histologic evidence of chronic suppurative cystitis and chronic, mild, lymphoplasmacytic pyelitis was present in all goats. Culture of renal tissue yielded bacterial growth in three of six goats, and culture of a swab of the urinary bladder mucosa yielded bacterial growth in all animals. Although clinical signs of ascending urinary tract infection were not observed in goats with patent stomata, urinary bladder marsupialization may result in ascending inflammation or infection. Based upon the results of this study, urinary bladder marsupialization should be recommended with caution as the primary procedure in clinical cases. / Master of Science

A 2,8-DHA Urolitíase é uma doença resultante de uma desordem hereditária que leva a deficiência de atividade da enzima APRT do grupo das PRTases. Até o momento, foram encontradas 18 mutações em pacientes, das quais 7 são missense. O presente trabalho dedica-se ao estudo funcional e estrutural dessas 7 mutações e da deleção &Delta;F173. Construções dos mutantes D65V, L110P, M136T, R67Q, R89Q, I112F e F173G foram obtidas no vetor pET-29a(+) e clonados em E. coli. Os protocolos de expressão e purificação foram estabelecidos, onde as enzimas foram obtidas após uma etapa cromatográfica na coluna CHTTM Hidroxiapatita Cerâmica no grau de pureza necessário. A única exceção foi o mutante I112F que se mostrou insolúvel durante a expressão. Um ensaio cinético acoplado através de kit para detecção de PPi foi padronizado e permitiu a caracterização cinética da hAPRT nativa e de seu mutantes. Os mutantes apresentaram perdas de eficiência de uma ordem de magnitude, com exceção do F173G, cujos valores foram comparáveis aos da hAPRT nativa. Ensaios de cristalização foram realizados para todos os mutantes e resultaram em cristais para os mutantes F173G, R67Q e R89Q. Esses cristais foram coletados na linha de Cristalografia de Macromoléculas 2 no LNLS. Os conjuntos de dados estão sendo analisados e as estruturas estão em fase final de refinamento. / 2,8-DHA Urolithiasis is a disease resulting from an inherited disorder that leads to deficiency of APRT, an enzyme of the PRTases group. So far, 18 mutations were found in patients, 7 of which are missense. This work is dedicated to the functional and structural study of these 7 mutations and the deletion &Delta;F173. Mutants constructions D65V, L110P, M136T, R67Q, R89Q, I112F and F173G were obtained in the vector pET-29a (+) and cloned in E. coli. The expression and purification protocols were established and the enzymes were obtained after 1 chromatographic step using the CHTTM Hydroxyapatite Ceramics column in the required purity. The only exception was the mutant I112F that resulted insoluble during expression. A coupled kinetic assay using PPi detection kit was standardized and allowed the kinetic characterization of native hAPRT and its mutants. The mutants had an efficiency loss of an order of magnitude, with the exception of F173G, whose values were comparable to those of native hAPRT. Crystallization trials were performed for all mutants and resulted in crystals for mutants F173G, R67Q and R89Q. These crystals were collected on Macromolecules Crystallography 2 line at LNLS. The datasets are being analyzed and the structures are in the final stages of refinement.

Made available in DSpace on 2014-06-11T19:31:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2003Bitstream added on 2014-06-13T20:21:52Z : No. of bitstreams: 1 amaro_crpr_dr_botfm.pdf: 783512 bytes, checksum: 77090e654f0b39da729c3d6aea6d8c92 (MD5) / A urolitíase é a terceira causa de afecção do trato urinário, com recorrência em torno de 50% em 5 anos após o primeiro cálculo. Afeta de 1 a 20% da população mundial, estando envolvidos fatores genéticos, epidemiológicos, diferenças geográficas, hábitos alimentares e, menos comumente, distúrbios hormonais. As investigações metabólicas têm grande importância na prevenção desta patologia. Os objetivos deste trabalho foram: diagnosticar os distúrbios metabólicos de pacientes litiásicos do Serviço de Litotripsia do Departamento de Urologia do Hospital das Clínicas da Faculdade Medicina de Botucatu-UNESP, estudar o sódio e o magnésio urinários como fatores de risco e conhecer aspectos epidemiológicos dessa população. PACIENTES E MÉTODOS: Foram estudados 158 pacientes com idade superior a 12 anos, no período de Fevereiro de 2000 a Dezembro de 2001. Foram incluidos pacientes com história e exames por imagem comprobatórios de no mínimo 2 cálculos, clearance de creatinina ≥ 60 ml/min e urocultura negativa. O protocolo consistiu da coleta não simultânea de 2 amostras de urina de 24h, para dosagens de Ca, P, AcU, Na, K, , Mg, Ox e Ci e dosagens séricas de glicemia, Ca, P, FA, AcU, Na, K, Cl, Mg, U, Cr; Realização do índice Ca/Cr urinários; índice Ca/Mg em urina 24h; índice de recorrência; pH urinário; prova de acidificação urinária. RESULTADOS: Os resultados obtidos foram: dos 158 pacientes estudados, 151 apresentaram pelo menos uma alteração metabólica. Destes,94(62,2%) apresentaram alteração metabólica isolada e 57(37,8%) alterações mistas. Os principais distúrbios encontrados foram: hipercalciúria(74,0%), hipocitratúria(37,3%), hiperoxalúria(24,1%), litíase úrica(20,2%), hiperparatireoidismo 1ario(1,8%) 2 ario (0,6%) e ATR(0,6%). A hipernatriúria esteve presente em 50% dos pacientes e a hipomagnesúria... / Urolithiasis is the third cause of urinary tract disease, with about 50% reoccurrence within five years of the first calculus. It affects between 1 and 20% of the world’s population, involving genetic and epidemiological factors, such as race; geographic differences; eating habits; and less commonly hormone disorders. Metabolic investigations are very important in preventing this pathology. The objectives of this work were to diagnose metabolic disorders in lithiasic patients from the Lithotripsy Service, Urology Department HC, FMB-UNESP, study sodium and magnesium as risk factors, and look at epidemiological aspects of this population. PATIENTS AND METHODS: We studied 158 patients all over 12 years old between February 2000 and December 2001. Patients were included with history and image exam confirmation of a minimum of 2 calculi, creatinine clearance ≥ 60 ml/min, and negative uroculture. The protocol consisted of non-simultaneous collect of two 24 hour urine samples for Ca, P, AcU, Na, K, Mg, Ox, and Ci levels, and serum levels of glycemia, Ca, P, FA, AcU, Na, K, Cl, Mg, U, and Cr; calculation of urine Ca/Cr index; 24h urine Ca/Mg index; recurrence index; urine pH; and urine acidity test. RESULTS: From the 158 patients studied, 151 presented at least one metabolic alteration; of these 94 were isolated alterations and 57 mixed. The main disorders were: hypercalciuria (74.0%); hypocitraturia (37.3%); hyperoxaluria (24.1%); uric acid lithiasis (20.2%); primary hyperparathyroidism (1.8%), secondary (0.6%); and ATR (0.6%). Hypernatriuria was present in 59% of patients and hypomagnesuria in 20.9%. Consumed water and urine volume did not show as risk factors. Hypernatriuria showed significant positive correlation in the male group only. There was no significant correlation between hypomagnesuria and...(Complete abstract click electronic address below)

A 2,8-DHA Urolitíase é uma doença resultante de uma desordem hereditária que leva a deficiência de atividade da enzima APRT do grupo das PRTases. Até o momento, foram encontradas 18 mutações em pacientes, das quais 7 são missense. O presente trabalho dedica-se ao estudo funcional e estrutural dessas 7 mutações e da deleção &Delta;F173. Construções dos mutantes D65V, L110P, M136T, R67Q, R89Q, I112F e F173G foram obtidas no vetor pET-29a(+) e clonados em E. coli. Os protocolos de expressão e purificação foram estabelecidos, onde as enzimas foram obtidas após uma etapa cromatográfica na coluna CHTTM Hidroxiapatita Cerâmica no grau de pureza necessário. A única exceção foi o mutante I112F que se mostrou insolúvel durante a expressão. Um ensaio cinético acoplado através de kit para detecção de PPi foi padronizado e permitiu a caracterização cinética da hAPRT nativa e de seu mutantes. Os mutantes apresentaram perdas de eficiência de uma ordem de magnitude, com exceção do F173G, cujos valores foram comparáveis aos da hAPRT nativa. Ensaios de cristalização foram realizados para todos os mutantes e resultaram em cristais para os mutantes F173G, R67Q e R89Q. Esses cristais foram coletados na linha de Cristalografia de Macromoléculas 2 no LNLS. Os conjuntos de dados estão sendo analisados e as estruturas estão em fase final de refinamento. / 2,8-DHA Urolithiasis is a disease resulting from an inherited disorder that leads to deficiency of APRT, an enzyme of the PRTases group. So far, 18 mutations were found in patients, 7 of which are missense. This work is dedicated to the functional and structural study of these 7 mutations and the deletion &Delta;F173. Mutants constructions D65V, L110P, M136T, R67Q, R89Q, I112F and F173G were obtained in the vector pET-29a (+) and cloned in E. coli. The expression and purification protocols were established and the enzymes were obtained after 1 chromatographic step using the CHTTM Hydroxyapatite Ceramics column in the required purity. The only exception was the mutant I112F that resulted insoluble during expression. A coupled kinetic assay using PPi detection kit was standardized and allowed the kinetic characterization of native hAPRT and its mutants. The mutants had an efficiency loss of an order of magnitude, with the exception of F173G, whose values were comparable to those of native hAPRT. Crystallization trials were performed for all mutants and resulted in crystals for mutants F173G, R67Q and R89Q. These crystals were collected on Macromolecules Crystallography 2 line at LNLS. The datasets are being analyzed and the structures are in the final stages of refinement.

Orientador: José Goldberg / Resumo: A urolitíase é a terceira causa de afecção do trato urinário, com recorrência em torno de 50% em 5 anos após o primeiro cálculo. Afeta de 1 a 20% da população mundial, estando envolvidos fatores genéticos, epidemiológicos, diferenças geográficas, hábitos alimentares e, menos comumente, distúrbios hormonais. As investigações metabólicas têm grande importância na prevenção desta patologia. Os objetivos deste trabalho foram: diagnosticar os distúrbios metabólicos de pacientes litiásicos do Serviço de Litotripsia do Departamento de Urologia do Hospital das Clínicas da Faculdade Medicina de Botucatu-UNESP, estudar o sódio e o magnésio urinários como fatores de risco e conhecer aspectos epidemiológicos dessa população. PACIENTES E MÉTODOS: Foram estudados 158 pacientes com idade superior a 12 anos, no período de Fevereiro de 2000 a Dezembro de 2001. Foram incluidos pacientes com história e exames por imagem comprobatórios de no mínimo 2 cálculos, clearance de creatinina ≥ 60 ml/min e urocultura negativa. O protocolo consistiu da coleta não simultânea de 2 amostras de urina de 24h, para dosagens de Ca, P, AcU, Na, K, , Mg, Ox e Ci e dosagens séricas de glicemia, Ca, P, FA, AcU, Na, K, Cl, Mg, U, Cr; Realização do índice Ca/Cr urinários; índice Ca/Mg em urina 24h; índice de recorrência; pH urinário; prova de acidificação urinária. RESULTADOS: Os resultados obtidos foram: dos 158 pacientes estudados, 151 apresentaram pelo menos uma alteração metabólica. Destes,94(62,2%) apresentaram alteração metabólica isolada e 57(37,8%) alterações mistas. Os principais distúrbios encontrados foram: hipercalciúria(74,0%), hipocitratúria(37,3%), hiperoxalúria(24,1%), litíase úrica(20,2%), hiperparatireoidismo 1ario(1,8%) 2 ario (0,6%) e ATR(0,6%). A hipernatriúria esteve presente em 50% dos pacientes e a hipomagnesúria...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Urolithiasis is the third cause of urinary tract disease, with about 50% reoccurrence within five years of the first calculus. It affects between 1 and 20% of the world's population, involving genetic and epidemiological factors, such as race; geographic differences; eating habits; and less commonly hormone disorders. Metabolic investigations are very important in preventing this pathology. The objectives of this work were to diagnose metabolic disorders in lithiasic patients from the Lithotripsy Service, Urology Department HC, FMB-UNESP, study sodium and magnesium as risk factors, and look at epidemiological aspects of this population. PATIENTS AND METHODS: We studied 158 patients all over 12 years old between February 2000 and December 2001. Patients were included with history and image exam confirmation of a minimum of 2 calculi, creatinine clearance ≥ 60 ml/min, and negative uroculture. The protocol consisted of non-simultaneous collect of two 24 hour urine samples for Ca, P, AcU, Na, K, Mg, Ox, and Ci levels, and serum levels of glycemia, Ca, P, FA, AcU, Na, K, Cl, Mg, U, and Cr; calculation of urine Ca/Cr index; 24h urine Ca/Mg index; recurrence index; urine pH; and urine acidity test. RESULTS: From the 158 patients studied, 151 presented at least one metabolic alteration; of these 94 were isolated alterations and 57 mixed. The main disorders were: hypercalciuria (74.0%); hypocitraturia (37.3%); hyperoxaluria (24.1%); uric acid lithiasis (20.2%); primary hyperparathyroidism (1.8%), secondary (0.6%); and ATR (0.6%). Hypernatriuria was present in 59% of patients and hypomagnesuria in 20.9%. Consumed water and urine volume did not show as risk factors. Hypernatriuria showed significant positive correlation in the male group only. There was no significant correlation between hypomagnesuria and...(Complete abstract click electronic address below) / Doutor

Objective: To assess effects of bisection nephrotomy on renal function, size, and morphology in cats over a period of 12 weeks. Study Design: Controlled, randomized, blinded experiment. Sample Population: Ten adult female cats. Methods: Glomerular filtration rate (GFR), as determined by quantitative renal scintigraphy using 99mTc-DTPA, urinalysis, urine culture, and sonographic measurement of renal size were performed pre-operatively. A left or right nephrotomy (five randomly assigned cats in each group) was performed. Total and individual kidney GFRs were determined in each cat 2, 28 and 84 days post-operatively. Both kidneys were measured sonographically 28 and 86 days postoperatively and an ultrasound-guided biopsy of each kidney was obtained 86 days postoperatively. Results: No significant differences in mean GFR and kidney size of the operated versus un-operated kidneys were observed at any time period. Individual GFR and renal size of all except one of the cats remained within normal limits. Two additional cats had evidence of transient ureteral obstruction in the immediate post-operative period. No significant histologic abnormalities were observed in any biopsy. Conclusions: Bisection nephrotomy in normal cats did not adversely affect renal function or morphology over the three month post-operative period. Clinical Relevance: Bisection nephrotomy can be safely performed in normal feline kidneys without causing a significant deleterious effect on renal function. Studies in animals with pre-existing renal insufficiency are needed to insure no adverse effects would occur in clinical cases where this surgical procedure is warranted. / Master of Science

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