RESUMO
Bladder cancer has a very high frequency of recurrence and therefore requires lifelong surveillance, traditionally consisting of serial cystoscopy and cytology. These tests are both invasive and expensive, with considerable inter-user and inter-institutional variability. In addition, the sensitivity of cytology in detecting low-grade tumors is low. Therefore, there has been active investigation into urinary biomarkers that can either supplement or supplant these tests. At this point there are only six urine-based tests that are FDA-approved in bladder cancer surveillance, but a wide variety of other biomarkers are being studied. In this review, we examine the natural history of bladder cancer as well as the rationale and performance of an ideal urinary biomarker. The authors describe the FDA-approved biomarkers such as Bladder Tumor Antigen, ImmunoCyt, Nuclear Matrix Protein-22, and Fluorescent In Situ Hybridization, as well as the most promising investigational tests (i.e., Urinary bladder cancer test, BLCA-1, BLCA-4, hyaluronic acid, hyaluronidase, Lewis X antigen, microsatellite analysis, Quanticyt, soluble Fas, Survivin, and telomerase). The biological foundation, methodologies, and diagnostic performance of the biomarkers are discussed. The characteristics of the biomarkers are compared to urine cytology. At this time, urine biomarkers are utilized in a variety of clinical situations but their role is not well defined. The goal of identifying an optimal marker that will replace cystoscopy and/or cytology is still ongoing.
Assuntos
Biomarcadores Tumorais/urina , Carcinoma de Células de Transição/urina , Vigilância da População/métodos , Neoplasias da Bexiga Urinária/urina , Adjuvantes Imunológicos/urina , Carcinoma de Células de Transição/diagnóstico , Inibidores de Cisteína Proteinase/urina , Cistoscopia , Proteína Ligante Fas/urina , Humanos , Ácido Hialurônico/urina , Hialuronoglucosaminidase/urina , Hibridização in Situ Fluorescente , Proteínas Inibidoras de Apoptose , Antígenos CD15/urina , Proteínas Associadas aos Microtúbulos/urina , Proteínas Nucleares/urina , Prognóstico , Sensibilidade e Especificidade , Survivina , Telomerase/urina , Neoplasias da Bexiga Urinária/diagnósticoRESUMO
Levamisole is a common adulterant in cocaine and has previously been associated with a variety of serious complications including multifocal inflammatory leukoencephalopathy (MIL). There have been several reports of MIL in patients taking cocaine and, though suspected, the presence of levamisole was not confirmed. We present a case of a 63-year-old woman presenting with stupor and spastic quadraparesis found to have urine positive for cocaine and levamisole. An MRI brain revealed innumerable FLAIR hyperintensities with restricted diffusion and incomplete ring-enhancement. This is the first case to confirm the presence of levamisole in a patient with MIL associated with cocaine use.
Assuntos
Adjuvantes Imunológicos/efeitos adversos , Transtornos Relacionados ao Uso de Cocaína/complicações , Transtornos Relacionados ao Uso de Cocaína/urina , Leucoencefalopatia Multifocal Progressiva/etiologia , Levamisol/efeitos adversos , Adjuvantes Imunológicos/urina , Transtornos Relacionados ao Uso de Cocaína/diagnóstico por imagem , Feminino , Humanos , Leucoencefalopatia Multifocal Progressiva/líquido cefalorraquidiano , Leucoencefalopatia Multifocal Progressiva/diagnóstico por imagem , Levamisol/urina , Imageamento por Ressonância Magnética , Pessoa de Meia-IdadeAssuntos
Adjuvantes Imunológicos/urina , Transtornos Relacionados ao Uso de Cocaína , Cocaína/química , Inibidores da Captação de Dopamina/química , Contaminação de Medicamentos , Levamisol/urina , Adjuvantes Imunológicos/efeitos adversos , Adjuvantes Imunológicos/farmacocinética , Agranulocitose/induzido quimicamente , Cocaína/urina , Inibidores da Captação de Dopamina/urina , Reações Falso-Negativas , Cromatografia Gasosa-Espectrometria de Massas , Hospitais Urbanos , Humanos , Imunoensaio , Leucoencefalopatias/induzido quimicamente , Levamisol/efeitos adversos , Levamisol/farmacocinética , Prevalência , Vasculite Leucocitoclástica Cutânea/induzido quimicamenteRESUMO
Human chorionic gonadotrophin (hCG) is a heterodimeric placental glycoprotein hormone required in pregnancy. In human pregnancy urine and in commercial hCG preparations (c-hCG) it occurs in a variety of forms, including breakdown products. Several reports have suggested modulation of the immune system by intact hormone, but such effects of breakdown products have not been reported. In a related article (Hum Immunol 62:1315, 2001), it is reported that a 400-2000 Dalton (Da) fraction from c-hCG and from human pregnancy urine inhibits Th1-mediated diabetes in NOD mice. The active component(s) were called natural (immuno)modulatory pregnancy factor(s) (NMPF). This study reports that a single treatment with the same low molecular weight NMPF fraction up to 24-h after high dose lipopolysaccharide (LPS) injection inhibited septic shock in mice. This counteracting effect of NMPF paralleled the downregulation of the effects of LPS on the production of macrophage migration inhibitory factor (MIF) by spleen cells, on the plasma level of liver aminotransferase, and on the expression of several splenic lymphocyte and macrophage surface markers. Based on the primary structure of the beta-chain of hCG a synthetic hexapeptide Valine-Leucin-Proline-Alanine-Leucine-Proline (VLPALP) was designed, which demonstrated it to have the same protective effects as the 400-2000 Da NMPF fraction. These results indicate a new strategy for the treatment of septic shock and the potential of therapeutic use of this synthetic oligopeptide.
Assuntos
Gonadotropina Coriônica Humana Subunidade beta/farmacologia , Oligopeptídeos/farmacologia , Choque Séptico/prevenção & controle , Adjuvantes Imunológicos/farmacologia , Adjuvantes Imunológicos/urina , Sequência de Aminoácidos , Animais , Gonadotropina Coriônica Humana Subunidade beta/química , Feminino , Humanos , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Lipopolissacarídeos/toxicidade , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Oligopeptídeos/química , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/farmacologia , Gravidez/imunologia , Gravidez/urina , Choque Séptico/tratamento farmacológico , Choque Séptico/etiologia , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/imunologiaRESUMO
Antineoplastons are naturally occurring cytodifferentiating agents. Chemically, they are medium and small sized peptides, amino acid derivatives and organic acids, which exist in blood, tissues and urine. Antineoplaston A-10 (3-phenylacetylamino-2,6-piperidinedione) is the first chemically identified antineoplaston. Previously we have shown a strong inverse association of urinary antineoplaston A-10 with breast cancer. This study is designed to evaluate neutrophil apoptosis in patients with breast cancer at time of diagnosis and to correlate urinary antineoplaston A-10 levels with neutrophil apoptosis and to describe the direct effect of A-10 in vitro on neutrophil apoptosis in breast cancer patients. The participants were patients with a histologically confirmed diagnosis of breast cancer. Only those cases without previous treatment for breast cancer were included. Neutrophil apoptosis was assessed in breast cancer patients both morphologically and by DNA fragmentation and studied relative to healthy controls. Antineoplaston A-10 was measured using high performance liquid chromatography in urine samples collected from the patients. Urine samples from normal women served as controls. Direct effect of antineoplaston A-10 on neutrophil apoptosis was tested in vitro after adding A-10 at a concentration of 10 ng/ml to the cellular suspensions of breast cancer patients. Non-treated samples served as controls. Significantly higher neutrophil apoptosis levels were detected among patients with breast cancer with a P value <0.001. Urinary antineoplaston A-10 level is significantly negatively correlated with high apoptosis levels (P<0.0001). In vitro, antineoplaston A-10 was found to inhibit significantly the neutrophil apoptosis with a P value <0.0001. These findings confirm the presence of immune defects among patients with breast cancer and such results should stimulate the development of new strategies to induce and augment immunity for the treatment of breast cancer. Antineoplaston A-10 may provide rational basis for designing trials to employ its immune modulatory potentials as adjuvant therapy in breast cancer patients.
Assuntos
Adjuvantes Imunológicos/farmacologia , Benzenoacetamidas , Neoplasias da Mama/imunologia , Neutrófilos/efeitos dos fármacos , Piperidonas/farmacologia , Adjuvantes Imunológicos/urina , Adulto , Idoso , Apoptose/efeitos dos fármacos , Apoptose/imunologia , Neoplasias da Mama/sangue , Neoplasias da Mama/urina , Células Cultivadas , Fragmentação do DNA , Feminino , Humanos , Pessoa de Meia-Idade , Neutrófilos/imunologia , Neutrófilos/patologia , Piperidonas/urinaRESUMO
Epidermal growth factor (EGF) and Tamm-Horsfall protein (THP) are synthesized in the kidneys by the distal tubular cells and excreted into urine. The urinary excretion of these peptides has been suggested as a potential index for distal tubular function. The urinary excretion rates of EGF and THP were examined in three groups of rats with increased renal excretion of urine: uninephrectomy, non-osmotic polyuria and diabetic osmotic polyuria. Twenty-four hour urine samples were obtained after 7, 14 and 21 days. The urinary volume per kidney was doubled in uninephrectomy when compared to controls. There was a seven-fold increase in urinary volume in rats with non-osmotic polyuria and diabetic osmotic polyuria, as compared to controls. Uninephrectomy, non-osmotic polyuria and diabetes all affected the urinary excretion of EGF and THP differently. The EGF excretion in uninephrectomized rats was 60-80% of that of the controls, whereas THP excretion was unchanged, indicating that EGF excretion varied with renal tissue mass. Non-osmotic polyuria caused a five-fold increase in THP excretion but no change in EGF excretion. THP excretion in the diabetic rats was increased three-fold after 21 days when compared to controls, whereas EGF excretion was decreased when expressed per kidney weight. Immunohistochemistry demonstrated that EGF and THP were colocalized in the thick ascending limbs of Henle's loops and distal tubules in all five groups of rats. In conclusion, the EGF excretion appears to follow renal tissue mass and seems independent of urinary volume, whereas THP excretion is dependent mainly on urinary volume. This has implications for the use of EGF and/or THP excretion rates as an indicator for distal tubular function.
Assuntos
Adjuvantes Imunológicos/urina , Diabetes Mellitus Experimental/urina , Fator de Crescimento Epidérmico/urina , Túbulos Renais Distais/metabolismo , Mucoproteínas/urina , Poliúria/urina , Animais , Modelos Animais de Doenças , Diurese , Ensaio de Imunoadsorção Enzimática , Feminino , Imuno-Histoquímica , Rim/metabolismo , Rim/patologia , Túbulos Renais Distais/patologia , Nefrectomia , Tamanho do Órgão , Concentração Osmolar , Ratos , Ratos Wistar , UromodulinaRESUMO
In a recent pharmacokinetic study, six human volunteers were exposed by inhalation to 10 ppm (14)C-D(4) for 1 h during alternating periods of rest and exercise. Octamethylcyclotetrasiloxane (D(4)) concentrations were determined in exhaled breath and blood. Total metabolite concentrations were estimated in blood, while the amounts of individual metabolites were measured in urine. Here, we use these data to develop a physiologically based pharmacokinetic (PBPK) model for D(4) in humans. Consistent with PBPK modeling efforts for D(4) in the rat, a conventional inhalation PBPK model assuming flow-limited tissue uptake failed to adequately describe these data. A refined model with sequestered D(4) in blood, diffusion-limited tissue uptake, and an explicit pathway for D(4) metabolism to short-chain linear siloxanes successfully described all data. Hepatic extraction in these volunteers, calculated from model parameters, was 0.65 to 0.8, i.e., hepatic clearance was nearly flow-limited. The decreased retention of inhaled D(4) seen in humans during periods of exercise was explained by altered ventilation/perfusion characteristics during exercise and a rapid approach to steady-state conditions. The urinary time course excretion of metabolites was consistent with a metabolic scheme in which sequential hydrolysis of linear siloxanes followed oxidative demethylation and ring opening. The unusual properties of D(4) (high lipophilicity coupled with high hepatic and exhalation clearance) lead to rapid decreases in free D(4) in blood. The success of D(4) PBPK models with a similar physiological structure in both humans and rats increases confidence in the utility of the model for predicting human tissue concentrations of D(4) and metabolites during inhalation exposures.
Assuntos
Adjuvantes Imunológicos/farmacocinética , Exercício Físico/fisiologia , Modelos Biológicos , Descanso/fisiologia , Siloxanas/farmacocinética , Adjuvantes Imunológicos/sangue , Adjuvantes Imunológicos/urina , Administração por Inalação , Algoritmos , Animais , Radioisótopos de Carbono , Relação Dose-Resposta a Droga , Gorduras/metabolismo , Humanos , Fígado/metabolismo , Masculino , Ratos , Reprodutibilidade dos Testes , Siloxanas/metabolismoRESUMO
A radioimmunoassay was developed for the measurement in human serum and urine of erythro-9-(2-hydroxy-3-nonyl)-hypoxanthine. Antisera were produced in rabbits by immunization with an erythro-9-(2-hydroxy-3-nonyl)-hypoxanthine hemisuccinate-bovine serum albumin conjugate. The competitive antigen was erythro-9-(2-hydroxy-3-nonyl)-hypoxanthine labeled with carbon-14 on the purine ring. Cross-reactivities were measured against three metabolites and the naturally occurring purine bases inosine and hypoxanthine. Sensitivity of the method was 1 ng/ml in serum and 10 ng/ml in urine. Precision at clinical levels was +/- 15% in serum at 2 ng/ml and +/- 3% in urine at 200 ng/ml.
Assuntos
Adjuvantes Imunológicos/análise , Hipoxantinas/análise , Adjuvantes Imunológicos/sangue , Adjuvantes Imunológicos/urina , Humanos , Hipoxantinas/sangue , Hipoxantinas/urina , Radioimunoensaio/métodosRESUMO
We describe an immunoblotting method for examining the electrophoretic properties of urinary Tamm-Horsfall protein. Using this method we tested frozen urine samples from a group of 37 thoroughly investigated recurrent idiopathic renal calcium stone formers and compared this with fresh urine from 19 non-stone forming laboratory staff. We found that there was a statistically significant different pattern of Tamm-Horsfall protein bands in the two sets of urines, with stone formers tending to have two bands and non-stone formers tending to have three bands. This could have been due to storage artefact and therefore a further group of 13 fresh urines from unselected renal stone formers was tested. A smaller proportion of these cases showed the two-band pattern, possibly because not all of this group were idiopathic calcium stone formers. This suggests that but does not prove that there is no significant storage artefact and that there may be an in vivo effect causing stone formation.
Assuntos
Adjuvantes Imunológicos/urina , Cálculos Renais/urina , Mucoproteínas/urina , Proteínas da Gravidez/urina , Adulto , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Masculino , Espectroscopia de Infravermelho com Transformada de Fourier , UromodulinaRESUMO
16alpha-Bromo-epiandrosterone (epiBr), a synthetic derivative of the natural hormone dehyroepiandrosterone (DHEA), was evaluated for its effects on feline immunodeficiency virus (FIV) infection in experimental cats. The rationale for this study was based on the ability of DHEA to significantly reduce the mortality to viral infections in mice. DHEA and epiBr also have demonstrable in vitro anti-viral activity for both HIV-1 and FIV. Preliminary pharmacokinetic studies in cats demonstrated that subcutaneously injected epiBr was rapidly absorbed, completely metabolized, and nontoxic. Metabolites were excreted in both urine and feces, with the latter having the most complex pattern of breakdown products. Cats were then divided into four groups; two groups were infected with FIV and two uninfected. Two groups, one infected and one uninfected were treated on 5 consecutive days of weeks 0, 4, 8, 12 and 16 with epiBr. The remaining two groups were mock treated with the drug vehicle alone. Treatment started 1 week prior to infection and extended for 4 weeks after infection. Cats were observed for 20 weeks post-FIV infection. Infected cats had identical decreases in blood neutrophil and lymphocyte counts following, regardless of whether they were treated with epiBr or vehicle alone. The CD4/CD8 T-cell ratio was decreased following FIV exposure, but was significantly more decreased for the epiBr treated animals from week 2 post-infection onward. CD4+ T cells were decreased in FIV-infected cats treated with epiBr compared to their untreated cohort, while CD8+ T cells tended to be higher in treated animals. FIV infected cats that were treated with epiBr had over one-log higher virus loads at week 2 post-infection than non-epiBr treated cohorts. In spite of this enhanced initial viremia, the subsequent levels of virus in the blood were significantly lower in epiBr treated versus untreated animals. EpiBr treated cats had significantly higher FIV-p24 antibody responses than control cats receiving vehicle alone, although primary and secondary antibody responses to a T-cell dependent non-FIV antigen, keyhole limpet hemocyanin (KLH), were unaffected. EpiBr treatment significantly decreased the expected FIV-induced suppression of IL-12 p40 mRNA levels in peripheral blood mononuclear cells (PBMCs) observed at weeks 4, 5, 8, 9 and 16 post-infection, but had no influence on FIV-induced changes in IL-4, IL-6, IL-10, IFN-gamma, MIP-1alpha and RANTES.
Assuntos
Adjuvantes Imunológicos/farmacologia , Androsterona/análogos & derivados , Androsterona/farmacologia , Síndrome de Imunodeficiência Adquirida Felina/tratamento farmacológico , Vírus da Imunodeficiência Felina/imunologia , Adjuvantes Imunológicos/farmacocinética , Adjuvantes Imunológicos/urina , Androsterona/farmacocinética , Androsterona/urina , Animais , Anticorpos Antivirais/biossíntese , Anticorpos Antivirais/sangue , Relação CD4-CD8/veterinária , Gatos , Citocinas/biossíntese , Citocinas/sangue , Citocinas/genética , Ensaio de Imunoadsorção Enzimática/veterinária , Fezes/química , Síndrome de Imunodeficiência Adquirida Felina/imunologia , Síndrome de Imunodeficiência Adquirida Felina/metabolismo , Síndrome de Imunodeficiência Adquirida Felina/virologia , Vírus da Imunodeficiência Felina/genética , Vírus da Imunodeficiência Felina/metabolismo , Interferon gama/genética , Interferon gama/imunologia , Masculino , Reação em Cadeia da Polimerase/veterinária , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Distribuição Aleatória , Organismos Livres de Patógenos Específicos , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Viremia/tratamento farmacológico , Viremia/imunologia , Viremia/veterináriaRESUMO
Dehydroepiandrosterone (DHEA) and androstenedione are weak androgens, which need conversion to more potent testosterone in order to enhance anabolic action. Consequences of oral dosing at 1 mg/kg on the urinary and plasma androgen profile of mare and gelding have been evaluated with an analytical method involving conjugate fractionation and selective hydrolysis, group separation, and quantitation by gas chromatography-mass spectrometry with selected ion monitoring of trimethylsilyl ethers. Peak levels of testosterone total conjugates in urine (range 300-6000 microg/L) were attained a few hours after dosing. Renal clearance was fast, so the testosterone detection period lasted only 20 to 33 h, the longest time being generated by androstenedione. The urinary testosterone/epitestosterone ratio for detection of exogenous testosterone in the mare was inoperative after DHEA administration because there was a concomitant increase of epitestosterone, which thereby acted as a masking agent. Androstanediols and androstenediols, as well as some 17-ketosteroids, were additional markers. A transient increase of circulating free testosterone has been evidenced, and this would support possible anabolic/androgenic action by supplementation with DHEA and androstenedione along the oral route.
Assuntos
Adjuvantes Imunológicos/sangue , Androstenodiona/sangue , Desidroepiandrosterona/sangue , Dopagem Esportivo , Cavalos/fisiologia , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/farmacocinética , Adjuvantes Imunológicos/urina , Administração Oral , Androstenodiona/administração & dosagem , Androstenodiona/farmacocinética , Androstenodiona/urina , Animais , Biomarcadores/análise , Desidroepiandrosterona/administração & dosagem , Desidroepiandrosterona/análise , Desidroepiandrosterona/farmacocinética , Desidroepiandrosterona/urina , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Masculino , Urinálise/veterináriaRESUMO
3-(2-Mercaptoethyl)quinazoline-2,4(1 H,3 H)-dione (1; AWD 100-041) is a substance with immunomodulating and immunorestorative activity. After p.o. administration in male Wistar rats at least 7 metabolites are formed and excreted in urine and faeces. The compounds were isolated and identified on the basis of UV and mass spectra. They are S-methylated structures in which sulfoxidation and ring-hydroxylation have been taken place. Four metabolites are also present as sulfate or glucuronide conjugates. The quantity ratio of the phase I to phase II metabolites amounts to 4:1. In the isolated perfused rat liver and rat hepatocyte culture 6 and 5 of the in vivo identified compounds are formed. The sequence of the metabolic pathways could be confirmed by in vitro experiments in which the incubation of synthetically prepared metabolites and the identification of generated biotransformation products were performed. In the lymphocyte and myeloma cell culture solely the disulfide of 1 is formed. After incubation of the S-methyl compound metabolites originate detectable also in vivo. Regarding the main ways of metabolism firstly 1 is attacked by methyltransferases forming the initial metabolite. After that oxidative processes take place leading to the formation of sulfoxides, sulfones as well as ring-hydroxylated compounds. A part of the ring-hydroxylated metabolites are conjugated.
Assuntos
Adjuvantes Imunológicos/farmacocinética , Quinazolinas/farmacocinética , Adjuvantes Imunológicos/urina , Animais , Biotransformação , Células Cultivadas , Fezes/química , Técnicas In Vitro , Fígado/metabolismo , Linfócitos/metabolismo , Masculino , Espectrometria de Massas , Quinazolinas/urina , Ratos , Ratos Wistar , Espectrofotometria Ultravioleta , Células Tumorais CultivadasRESUMO
BALB/c mice with pulmonary tuberculosis develop a T helper cell type 1 response that peaks at 3 weeks, temporarily controlling bacterial growth. Then bacterial proliferation recommences, accompanied by increasing interleukin (IL)-4 levels and decreasing interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and inducible nitric oxide synthase (iNOS) levels. These changes mimic those in the human disease. In a previous study, administration of dehydroepiandrosterone (DHEA) beginning on day 60 after infection reversed these changes and protected the mice. However, DHEA is suboptimal for human use, partly because it is readily metabolized into sex steroids. 16alpha-Bromoepiandrosterone (EpiBr; 16alpha -bromo-5alpha -androstan-3beta-ol-17-one) is a synthetic adrenal steroid derivative that does not enter sex steroid pathways. In the present study, when tuberculous BALB/c mice were treated with EpiBr 3 times/week beginning on day 60, inhibition of bacterial proliferation and increased expression of TNF-alpha, IFN-gamma, and iNOS were observed, although decreased expression of IL-4 was also observed. Moreover, when given as an adjunct to conventional chemotherapy, EpiBr enhanced bacterial clearance. Trials for the use of EpiBr in the treatment of human tuberculosis are now justified.
Assuntos
Adjuvantes Imunológicos/farmacologia , Androsterona/análogos & derivados , Androsterona/farmacologia , Quimioterapia Adjuvante , Células Th1/efeitos dos fármacos , Tuberculose Pulmonar/imunologia , Adjuvantes Imunológicos/farmacocinética , Adjuvantes Imunológicos/urina , Animais , Ativação Linfocitária/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Mycobacterium tuberculosis/imunologia , Células Th1/imunologia , Tuberculose Pulmonar/metabolismo , Tuberculose Pulmonar/patologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
INTRODUCTION: Oxalate in urine can cause tubular cellular damage by the production of free radicals. Then, cell death and cellular debris may promote the retention of calcium oxalate crystals and finally the formation of stones. The two most abundant urinary proteins, Tamm-Horsfall protein (THP) and albumin, were tested for the effects of antioxidants. MATERIALS AND METHODS: By using xanthine-xanthine oxidase reaction, purified THP and albumin were tested for the inhibitory effect. OD(295) was used as a spectrophotometric method to measure the production of uric acid during the reaction. RESULTS AND CONCLUSIONS: Both proteins can inhibit the reaction of xanthine oxidase on xanthine, although the effect was decreased after enzymatic deglycosylation of sialic acid. Albumin has an IC(50) of 10.7 nM in native condition and 11.9 nM after deglycosylation, whereas THP has 69.6 nM in native condition and 102.0 nM in deglycosylated condition. The data indicates that THP and albumin have an antioxidant effect. Sialic acid in THP has partly an inhibitory effect and is associated with calcium oxalate formation. Studies have indicated that further investigation of the role of free radicals in the formation of urolithiasis and of sialic acid in protein function is needed.
Assuntos
Adjuvantes Imunológicos/metabolismo , Albuminas/metabolismo , Oxalato de Cálcio/metabolismo , Radicais Livres/antagonistas & inibidores , Mucoproteínas/metabolismo , Xantina Oxidase/antagonistas & inibidores , Adjuvantes Imunológicos/urina , Cristalização , Eletroforese em Gel de Poliacrilamida , Humanos , Masculino , Mucoproteínas/urina , Cálculos Urinários/fisiopatologia , Uromodulina , Xantina/metabolismoRESUMO
The purpose of the present study was to quantify the binding affinity between Tamm-Horsfall protein (THP) and complement 1q (C1q) using ELISA and a resonant mirror biosensor. In ELISA, immobilized THP was incubated with soluble C1q under both low and physiological ionic-strength conditions. Tamm-Horsfall protein bound C1q with an equilibrium dissociation constant (KD) of 1.9 +/- 0.6 nmol/L in low ionic-strength Tris buffers (20 mmol/L NaCl, pH 7.5) and with a lower affinity (KD of 13.4 +/- 4.7 nmol/L) in physiological-strength Tris buffers (154 mmol/L NaCl, pH 7.5). A resonant mirror biosensor, which monitors binding events in real-time, was used to quantify the KD of this reaction, as well as to estimate the kinetic parameters. In these studies, THP and C1q bound with an association rate constant, kass, of 1.25 x 105 L/mol per s and a dissociation rate constant, kdiss, of 0.002-0.005/s. The calculated KD for the THP/C1q binding in low ionic-strength buffers was higher (averages of 10-15 nmol/L) than that obtained by the ELISA, while physiological ionic-strength buffers still reduced the affinity of this binding by an order of magnitude. In conclusion, THP consistently bound C1q with high affinity using several techniques. At least a portion of this interaction involved electrostatic events, as demonstrated by the influence of ionic strength on the binding affinity.
Assuntos
Adjuvantes Imunológicos/metabolismo , Complemento C1q/metabolismo , Ensaio de Imunoadsorção Enzimática , Mucoproteínas/metabolismo , Adjuvantes Imunológicos/urina , Adulto , Técnicas Biossensoriais/métodos , Feminino , Humanos , Cinética , Masculino , Pessoa de Meia-Idade , Mucoproteínas/urina , Concentração Osmolar , Ligação Proteica , Reprodutibilidade dos Testes , Eletricidade Estática , UromodulinaRESUMO
Tamm-Horsfall glycoproteins (THPs) from healthy probands and a majority of recurrent calcium oxalate renal stone formers reveal different physicochemical properties when analyzed using isoelectric focusing (IEF). The pI values of THPs from healthy probands are approximately 3.5 while THPs from recurrent renal stone formers have pI values of between 4.5 and 6. The two groups of THPs exhibit completely different protein patterns. The differences in IEF analysis allow differentiation between THPs from healthy probands and recurrent calcium oxalate stone formers and may possibly be used as a simple diagnostic method for the recognition of recurrent calcium oxalate renal stone formers.
Assuntos
Adjuvantes Imunológicos/análise , Oxalato de Cálcio/análise , Mucoproteínas/análise , Cálculos Urinários/química , Cálculos Urinários/etiologia , Adjuvantes Imunológicos/urina , Oxalato de Cálcio/urina , Densitometria , Detergentes , Géis , Glicoproteínas/análise , Glicoproteínas/urina , Humanos , Focalização Isoelétrica , Mucoproteínas/urina , Ureia , UromodulinaRESUMO
This paper describes a simple high-performance liquid chromatographic method for the determination of PGT/1A (3-L-pyroglutamyl-L-thiazolidine-4-carboxylic acid), a new immunostimulating drug, in plasma and urine. The column was packed with LiChrospher-NH2 (5 microns), the mobile phase was 0.02 M monobasic potassium phosphate (pH 3.2 with concentrated phosphoric acid)-acetonitrile (25:75, v/v), the flow-rate was 1.2 ml/min, the detection wavelength was 210 nm and the apparatus was a Varian Model 5000. Plasma (1 ml) was added to 1.2 ml of acetonitrile and the supernatant injected; the urine was diluted 1:5. The retention time of PGT/1A was 9.4 min in plasma and 9.9 min in urine. The method was validated for recovery, accuracy and reproducibility. The results after intravenous injection in twelve volunteers are also given.
Assuntos
Adjuvantes Imunológicos/sangue , Cromatografia Líquida de Alta Pressão/métodos , Compostos de Potássio , Ácido Pirrolidonocarboxílico/análogos & derivados , Tiazóis/sangue , Acetonitrilas/administração & dosagem , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/urina , Adulto , Humanos , Injeções Intravenosas , Masculino , Fosfatos/administração & dosagem , Potássio/administração & dosagem , Ácido Pirrolidonocarboxílico/administração & dosagem , Ácido Pirrolidonocarboxílico/sangue , Ácido Pirrolidonocarboxílico/urina , Tiazóis/administração & dosagem , Tiazóis/urina , TiazolidinasRESUMO
An high-performance liquid chromatographic analysis of PCF 39, N2-[5-(hypoxanthin-9-yl)pentyloxycarbonyl]-L-arginine, with ultraviolet detection, has been devised and validated. The main pharmacokinetic results encountered for rats treated intravenously with PCF 39 at a dose of 100 mg/kg are described.
Assuntos
Adjuvantes Imunológicos/farmacocinética , Arginina/análogos & derivados , Hipoxantinas/farmacocinética , Adjuvantes Imunológicos/sangue , Adjuvantes Imunológicos/urina , Animais , Arginina/sangue , Arginina/farmacocinética , Arginina/urina , Cafeína/sangue , Cafeína/urina , Fenômenos Químicos , Química , Cromatografia Líquida de Alta Pressão , Hipoxantinas/sangue , Hipoxantinas/urina , Masculino , Ratos , Ratos EndogâmicosRESUMO
UNLABELLED: This study aimed assess function of renal tubules in patients undergoing conditioning regimen before bone marrow transplantation. The examined group comprised 19 patients. 13 of them underwent autologuous bone marrow transplantation (ABMT), or autologuous peripheral blood stem cells transplantation (APBSCT). These patients were treated with Cyclophosphamide (Cy) (120 mg/kg), Etoposide (1.6-1.8 g/m2) and Carmustine (400-450 mg/m2). The remaining 6 patients underwent allogenic bone marrow transplantation (BMT). They were treated with Cy 200 mg/kg or with Cy 120 mg/kg and Busulfan 16 mg/kg (doses given are the total amount or respective drugs were administered during the whole conditioning regimen). Urinary excretion of beta-2MB and THP was assessed a) before the conditioning regimen was started, b) one day before completion of it, and c) one day before bone marrow transplantation. In ABMT/APBSCT patients urinary excretion of beta-2-MG was significantly higher before and during conditioning regimen as compared with baseline value (3309 +/- 1123.7 vs 3919 +/- 1417.8 vs 246.7 +/- 50.3 mg/24h, respectively, p < 0.05). Urinary excretion of THP in these patients was significantly higher only during the conditioning regimen (90.6 +/- 14.3 vs. 30.4 +/- 6.24 mg/24h, p < 0.0002). In BMT patients conditioning regimen was followed by similar but less marked changes of urinary excretion of beta-2-MG and THP as compared with ABMT/APBSCT patients. CONCLUSIONS: 1. Conditioning regimen do influence significantly function of proximal and distal renal tubules. 2. The extent of disturbed function of renal tubules seems to depend on kind of medication that is given during the conditioning regimen.