RESUMEN
BACKGROUND: Contemporary studies have discredited the methods used to exclude urinary tract infection (UTI) when treating overactive bladder (OAB). Thus we must revisit the OAB phenotype to check that UTI has not been overlooked. AIMS: To examine the differences in urinary cytokines IL6 and lactoferrin in OAB patients compared to controls, with references to microscopy of urine and enhanced quantitative urine culture. METHODS: A blinded, prospective cohort study with normal controls using six repeated measures, achieved two-monthly, over 12 months. RESULTS: The differences between patients and controls in urine IL6 (F = 49.0, p < .001) and lactoferrin (F = 228.5, p < .001) were significant and of a magnitude to have clinical implications. These differences were for lactoferrin correlated to symptoms (9.3, p = .003); for both to pyuria (IL6 F = 66.2, p < .001, Lactoferrin F = 73.9, p < .001); and for IL6 microbial abundance (F = 5.1, p = .024). The pathological markers had been missed by urinary dipsticks and routine MSU culture. CONCLUSION: The OAB phenotype may encompass patients with UTI that is being overlooked because of the failure of standard screening methods.
Asunto(s)
Interleucina-6/orina , Lactoferrina/orina , Vejiga Urinaria Hiperactiva/orina , Anciano , Femenino , Humanos , Inflamación/etiología , Inflamación/orina , Persona de Mediana Edad , Estudios Prospectivos , Método Simple Ciego , Vejiga Urinaria Hiperactiva/complicacionesRESUMEN
Urothelial carcinoma of the bladder (UCB) is potentially life-threatening; therefore, we aimed to discover a novel urine biomarker for diagnosis and prognostication of UCB. This is a retrospective case-control study. Exploration of a new biomarker using urine from 20 UCB patients in the present study revealed that urinary level of lactoferrin (LF), a multifunctional glycoprotein released from neutrophils, was higher in 11 of 15 with invasive/high-grade UCB than 5 with non-invasive one, and 2 healthy adults. We therefore focused on LF and assessed the value of urine LF normalized by urine creatinine concentration (LF/Cr) using an enzyme-linked immunosorbent assay. Diagnostic performance of urine LF/Cr was examined using urine from 92 patients with primary (newly diagnosed) untreated UCB and 166 controls without UCB, including 62 patients with pyuria, and 104 subjects without pyuria consisting of 84 patients and 20 healthy adults. However, the diagnostic accuracies were accompanied by the risk of bias. In 92 primary UCB patients, both pyuria and tumor-infiltrating neutrophils (TINs) were independent predictors for urine LF/Cr. In contrast, TINs or urine LF/Cr were independent predictors for invasive histology, whereas pyuria was not. In terms of prognostication, urine LF/Cr and nodal metastasis were independent predictors of disease-specific survival in 22 patients with muscle-invasive bladder cancer, characterized by a high mortality rate, in the Cox proportional hazards model. In conclusion, urine LF/Cr linked to TINs was a predictor of both invasive histology and prognosis in UCB. Urine LF/Cr is a potential biomarker reflecting the degree of malignancy in UCB.
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Biomarcadores/orina , Carcinoma/orina , Lactoferrina/orina , Neoplasias de la Vejiga Urinaria/orina , Urotelio/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Proliferación Celular , Niño , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , Metástasis de la Neoplasia , Neutrófilos/metabolismo , Pronóstico , Estándares de Referencia , Estudios Retrospectivos , Vejiga Urinaria/patología , Adulto JovenRESUMEN
BACKGROUND: Early administration of colostrum may provide preterm infants with immune components. Previous studies illustrating the effects of oral colostrum (OC) have been confounded by the coincidence of enteral feedings. OBJECTIVE: To quantify OC absorption, as measured by urinary sIgA and lactoferrin, in preterm infants prior to enteral feedings. MATERIALS AND METHODS: Colostrum was obtained from mothers delivering infants ≤32 weeks and ≤1500 g. sIgA and lactoferrin were measured in infant urine, and microflora in saliva and tracheal aspirates were characterized. RESULTS: Urinary sIgA and lactoferrin were significantly greater in infants receiving OC by syringe compared to swab (p < 0.002). Urinary sIgA correlated with the total number of doses in 72 h (R2 = 43%, p < 0.01). CONCLUSIONS: Administration of OC by syringe and higher cumulative dose are associated with increased absorption of sIgA and lactoferrin, and early dosing may contribute to a more diverse tracheal microbiome.
Asunto(s)
Calostro/inmunología , Inmunoglobulina A Secretora/orina , Recien Nacido Prematuro/inmunología , Recién Nacido de muy Bajo Peso/inmunología , Lactoferrina/orina , Administración Oral , Análisis de Varianza , Humanos , Recién Nacido , Recien Nacido Prematuro/orina , Microbiota , Boca/microbiología , Mucosa Bucal , Proyectos Piloto , Tráquea/microbiologíaRESUMEN
Urinary tract infection (UTI) is a prominent global health care burden. Although UTI is readily treated with antibiotics in healthy adults, complicated cases in immune-compromised individuals and the emerging antibiotic resistance of several uropathogens have accelerated the need for new treatment strategies. Here, we surveyed the composition of urinary exosomes in a mouse model of uropathgenic Escherichia coli (UPEC) UTI to identify specific urinary tract defense constituents for therapeutic development. We found an enrichment of the iron-binding glycoprotein lactoferrin in the urinary exosomes of infected mice. In subsequent in vitro studies, we identified human bladder epithelial cells as a source of lactoferrin during UPEC infection. We further established that exogenous treatment with human lactoferrin (hLf) reduces UPEC epithelial adherence and enhances neutrophil antimicrobial functions including bacterial killing and extracellular trap production. Notably, a single intravesicular dose of hLf drastically reduced bladder bacterial burden and neutrophil infiltration in our murine UTI model. We propose that lactoferrin is an important modulator of innate immune responses in the urinary tract and has potential application in novel therapeutic design for UTI.
Asunto(s)
Infecciones por Escherichia coli/inmunología , Exosomas/metabolismo , Lactoferrina/orina , Neutrófilos/inmunología , Vejiga Urinaria/inmunología , Infecciones Urinarias/inmunología , Escherichia coli Uropatógena/fisiología , Animales , Modelos Animales de Enfermedad , Trampas Extracelulares/metabolismo , Femenino , Humanos , Inmunidad Innata , Huésped Inmunocomprometido , Hierro/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Vejiga Urinaria/microbiologíaRESUMEN
BACKGROUND: Urinary tract infection (UTI) is a common infection that poses a substantial healthcare burden, yet its definitive diagnosis can be challenging. There is a need for a rapid, sensitive and reliable analytical method that could allow early detection of UTI and reduce unnecessary antibiotics. Pathogen identification along with quantitative detection of lactoferrin, a measure of pyuria, may provide useful information towards the overall diagnosis of UTI. Here, we report an integrated biosensor platform capable of simultaneous pathogen identification and detection of urinary biomarker that could aid the effectiveness of the treatment and clinical management. METHODOLOGY/PRINCIPAL FINDINGS: The integrated pathogen 16S rRNA and host lactoferrin detection using the biosensor array was performed on 113 clinical urine samples collected from patients at risk for complicated UTI. For pathogen detection, the biosensor used sandwich hybridization of capture and detector oligonucleotides to the target analyte, bacterial 16S rRNA. For detection of the protein biomarker, the biosensor used an analogous electrochemical sandwich assay based on capture and detector antibodies. For this assay, a set of oligonucleotide probes optimized for hybridization at 37°C to facilitate integration with the immunoassay was developed. This probe set targeted common uropathogens including E. coli, P. mirabilis, P. aeruginosa and Enterococcus spp. as well as less common uropathogens including Serratia, Providencia, Morganella and Staphylococcus spp. The biosensor assay for pathogen detection had a specificity of 97% and a sensitivity of 89%. A significant correlation was found between LTF concentration measured by the biosensor and WBC and leukocyte esterase (p<0.001 for both). CONCLUSION/SIGNIFICANCE: We successfully demonstrate simultaneous detection of nucleic acid and host immune marker on a single biosensor array in clinical samples. This platform can be used for multiplexed detection of nucleic acid and protein as the next generation of urinary tract infection diagnostics.
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Técnicas Biosensibles/métodos , Técnicas Electroquímicas/instrumentación , Técnicas Electroquímicas/métodos , Ácidos Nucleicos/orina , Proteínas/análisis , Infecciones Urinarias/diagnóstico , Infecciones Urinarias/orina , Humanos , Lactoferrina/orina , Hibridación de Ácido Nucleico , Sondas de Oligonucleótidos/metabolismo , ARN Ribosómico 16S/genética , Sensibilidad y Especificidad , TemperaturaRESUMEN
UNLABELLED: Own mother's colostrum is rich in cytokines and other immune agents that may stimulate oropharyngeal-associated lymphoid tissue if administered oropharyngeally to extremely low-birth-weight (ELBW) infants during the first days of life when enteral feeding is contraindicated. However, the safety and feasibility of the oropharyngeal route for the administration of colostrum have not been determined. PURPOSE: To determine the safety of oropharyngeal administration of own mother's colostrum to ELBW infants in first days of life. A secondary purpose was to investigate the feasibility of (1) delivering this intervention to ELBW infants in the first days of life and (2) measuring concentrations of secretory immunoglobulin A and lactoferrin in tracheal aspirate secretions and urine of these infants. SUBJECTS: Five ELBW infants (mean birth weight and gestational age = 657 g and 25.5 weeks, respectively). DESIGN: Quasi-experimental, 1 group, pretest-posttest design. METHODS: Subjects received 0.2 mL of own mother's colostrum administered oropharyngeally every 2 hours for 48 consecutive hours, beginning at 48 hours of life. Concentrations of secretory immunoglobulin A and lactoferrin were measured in tracheal aspirates and urine of each subject at baseline, at the completion of the intervention and again 2 weeks later. RESULTS: All infants completed the entire treatment protocol, each receiving 24 treatments. A total of 15 urine specimens were collected and 14 were sufficient in volume for analysis. A total of 15 tracheal aspirates were collected, but only 7 specimens (47%) were sufficient in volume for analysis. There was wide variation in concentrations of secretory immunoglobulin A and lactoferrin in urine and tracheal aspirates among the 5 infants; however, several results were outside the limits of assay detection. All infants began to suck on the endotracheal tube during the administration of colostrum drops. Oxygen saturation measures remained stable or increased slightly during each of the treatment sessions. There were no episodes of apnea, bradycardia, hypotension, or other adverse effects associated with the administration of colostrum. CONCLUSIONS: Oropharyngeal administration of own mother's colostrum is easy, inexpensive, and well-tolerated by even the smallest and sickest ELBW infants. Future research should continue to examine the optimal procedure for measuring the direct immune effects of this therapy, as well as the clinical outcomes such as infections, particularly ventilator-associated pneumonia.
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Calostro/inmunología , Nutrición Enteral/métodos , Inmunoglobulina A Secretora/metabolismo , Lactoferrina/metabolismo , Administración Oral , Secreciones Corporales/metabolismo , Nutrición Enteral/efectos adversos , Femenino , Humanos , Inmunoglobulina A Secretora/orina , Recien Nacido con Peso al Nacer Extremadamente Bajo , Recién Nacido , Recien Nacido Prematuro , Lactoferrina/orina , Masculino , Proyectos Piloto , Tráquea/inmunologíaRESUMEN
Urine is the most abundant and easily accessible of all body fluids and provides an ideal route for non-invasive diagnosis of human diseases, particularly of the urinary tract. Electrochemical biosensors are well suited for urinary diagnostics due to their excellent sensitivity, low-cost, and ability to detect a wide variety of target molecules including nucleic acids and protein biomarkers. We report the development of an electrochemical immunosensor for direct detection of the urinary tract infection (UTI) biomarker lactoferrin from infected clinical samples. An electrochemical biosensor array with alkanethiolate self-assembled monolayer (SAM) was used. Electrochemical impedance spectroscopy was used to characterize the mixed SAM, consisted of 11-mercaptoundecanoic acid and 6-mercapto-1-hexanol. A sandwich amperometric immunoassay was developed for detection of lactoferrin from urine, with a detection limit of 145 pg/ml. We validated lactoferrin as a biomarker of pyuria (presence of white blood cells in urine), an important hallmark of UTI, in 111 patient-derived urine samples. Finally, we demonstrated multiplex detection of urinary pathogens and lactoferrin through simultaneous detection of bacterial nucleic acid (16S rRNA) and host immune response protein (lactoferrin) on a single sensor array. Our results represent first integrated sensor platform capable of quantitative pathogen identification and measurement of host immune response, potentially providing clinical diagnosis that is not only more expeditious but also more informative than the current standard.
Asunto(s)
Técnicas Biosensibles/instrumentación , Conductometría/instrumentación , Lactoferrina/orina , Infecciones Urinarias/diagnóstico , Infecciones Urinarias/orina , Diseño de Equipo , Análisis de Falla de Equipo , HumanosRESUMEN
BACKGROUND: Because of the risk of performing renal biopsies in children with co-morbid conditions, we carried out this study to identify candidate protein biomarkers in the urine of HIV-infected children with renal disease. DESIGN, SETTING, PARTICIPANTS & MEASUREMENTS: Urine samples from HIV-infected children with biopsy proven HIV-nephropathy (HIVAN; n = 4), HIV-associated Hemolytic Uremic Syndrome (HIV-HUS; n = 2), or no renal disease (n = 3) were analyzed by two-dimensional electrophoresis (2-DE) and proteomic methods. Positive findings were confirmed in HIV-infected children with (n = 20) and without (n = 10) proteinuria using commercially available assays. RESULTS: By 2-DE analysis, a single urine marker was not sufficient to distinguish children with HIVAN from the others. High urine levels of beta(2)-microglobulin and retinol-binding protein (RBP) suggested the presence of tubular injury. In addition, we found elevated urine levels of iron and the iron-related proteins, transferrin, hemopexin, haptoglobin, lactoferrin, and neutrophil gelatinase-associated lipocalin (NGAL), in children with HIVAN and HIV-HUS. Furthermore, we detected a significant accumulation of iron in the urine and kidneys of HIV-transgenic (Tg) rats with renal disease. CONCLUSION: These findings suggest that iron and iron-related proteins might be promising candidate urine biomarkers to identify HIV-infected children at risk of developing HIVAN and HIV-HUS. Moreover, based on the results of previous studies, we speculate that the release or accumulation of iron in the kidney of HIV-infected children may contribute to the rapid progression of their renal disease, and could become a new therapeutic target against HIVAN and HIV-HUS.
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Nefropatía Asociada a SIDA/orina , Proteínas Sanguíneas/orina , Infecciones por VIH/virología , VIH-1/patogenicidad , Síndrome Hemolítico-Urémico/orina , Proteinuria/orina , Nefropatía Asociada a SIDA/patología , Nefropatía Asociada a SIDA/virología , Proteínas de Fase Aguda/orina , Animales , Biomarcadores/orina , Biopsia , Estudios de Casos y Controles , Modelos Animales de Enfermedad , Infecciones por VIH/complicaciones , Infecciones por VIH/patología , Infecciones por VIH/orina , VIH-1/genética , Haptoglobinas/orina , Síndrome Hemolítico-Urémico/patología , Síndrome Hemolítico-Urémico/virología , Hemopexina/orina , Humanos , Hierro/orina , Lactoferrina/orina , Lipocalina 2 , Lipocalinas/orina , Valor Predictivo de las Pruebas , Proteinuria/virología , Proteínas Proto-Oncogénicas/orina , Ratas , Ratas Transgénicas , Factores de Tiempo , Transferrina/orinaRESUMEN
Lactoferrin is an iron binding glycoprotein found in the 2ry granules of PMN. In order to determine the usefulness of such marker for neutrophilic activity in differentiating cases suffering from amoebic and bacillary dysentery, Schistosoma and bacterial UTI infections, we examined stool and urine specimens using anti-lactoferrin antibodies (lactoferrin latex agglutination test: LFLA), compared with different standard gold techniques. Our results demonstrated that cases with either shigllosis or UTI revealed a high lactoferrin titer which was positively correllated with the number of PMN. In addition cases with Entamoeba histolytica or S. haematobium were characterized by relatively lower inflammatory process as expressed by mild lactoferrin titer which was also correlated with the PMN count. In addition, the findings of the present work indicated that LFLA was sensitive and specific when used alone and its sensitivity was augmented after coupling with other simple indirect methods of diagnosis. In conclusion, results described the reliability of using LFLA as a simple, rapid, sensitive method in differentiating, certain parasitic from bacterial diseases.
Asunto(s)
Disentería Amebiana/diagnóstico , Disentería Bacilar/diagnóstico , Entamoeba histolytica/crecimiento & desarrollo , Lactoferrina/análisis , Schistosoma haematobium/crecimiento & desarrollo , Esquistosomiasis Urinaria/diagnóstico , Shigella/inmunología , Animales , Diagnóstico Diferencial , Disentería Amebiana/orina , Disentería Bacilar/orina , Heces/química , Humanos , Lactoferrina/orina , Esquistosomiasis Urinaria/orinaRESUMEN
Bovine lactoferrin (LF), which is an iron-binding glycoprotein in milk, was administered orally to groups of 12 males and 12 female rats at dose levels of 200, 600 and 2000mg/kg/day once daily for 13 weeks and its toxicity on repeated administration was examined. Throughout the administration period, there were no deaths caused by administration of the test compound, nor were there any adverse effects noted in the general condition of the animals. The study findings concerning body weight and food consumption, ophthalmology, urinalysis including water consumption, haematology, blood chemistry, necropsy, organ weights and histopathology revealed that there were no apparent changes due to administration of LF. Therefore, the level of LF at which no adverse effect was observed was considered to be 2000mg/kg/day for both sexes.
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Lactoferrina/toxicidad , Administración Oral , Animales , Antiinfecciosos/administración & dosificación , Antiinfecciosos/toxicidad , Antiinfecciosos/orina , Peso Corporal/efectos de los fármacos , Bovinos , Ingestión de Alimentos/efectos de los fármacos , Femenino , Lactoferrina/administración & dosificación , Lactoferrina/orina , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
The usefulness of the measurement of urinary lactoferrin (LF) released from polymorphonuclear leukocytes and of an immunochromatography test strip devised for measuring urinary LF for the simple and rapid diagnosis of urinary tract infections (UTI) was evaluated. Urine specimens were collected from apparently healthy persons and patients diagnosed as suffering from UTI. In the preliminary study, the LF concentrations in 121 normal specimens and 88 specimens from patients (60 with UTI) were quantified by an enzyme-linked immunosorbent assay. The LF concentration was 3,300.0 +/- 646.3 ng/ml (average +/- standard error of the mean) in the specimens from UTI patients, whereas it was 30.4 +/- 2.7 ng/ml and 60.3 +/- 14.9 ng/ml in the specimens from healthy persons and the patients without UTI, respectively. Based on these results, a 200-ng/ml LF concentration was chosen as the cutoff value for negativity. Each urine specimen was reexamined with the newly devised immunochromatography (IC) test strip to calculate the indices of efficacy. Based on the cutoff value, it was calculated that the sensitivity, specificity, and positive and negative predictive values of the IC test were 93.3, 89.3, 86.2, and 94.9%, respectively, compared with the results of the microscopic examination of the urine specimens for the presence of leukocytes. The respective indices for UTI were calculated as 95.0, 92.9, 89.7, and 96.6%. The tests were completed within 10 min. These results indicated that urine LF measurement with the IC test strip provides a useful tool for the simple and rapid diagnosis of UTI.
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Lactoferrina/orina , Infecciones Urinarias/diagnóstico , Adulto , Anticuerpos , Anticuerpos Monoclonales , Biomarcadores/orina , Cromatografía , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Lactoferrina/sangre , Masculino , Persona de Mediana Edad , Neutrófilos , Tiras Reactivas , Valores de Referencia , Reproducibilidad de los Resultados , Infecciones Urinarias/orina , Orina/citologíaRESUMEN
The authors propose a highly effective method for isolation of lactoferrin from female colostrum. Lactoferrin is measured by the sandwich enzyme immunoassay. Its concentrations in the serum, urine, amniotic and cerebrospinal fluid in health are measured. The advantages of the method for lactoferrin purification and the practical significance of this protein are discussed.
Asunto(s)
Técnicas para Inmunoenzimas , Lactoferrina/análisis , Calostro/química , Femenino , Humanos , Lactoferrina/sangre , Lactoferrina/líquido cefalorraquídeo , Lactoferrina/orinaRESUMEN
Intact (i.e., 78-kDa) lactoferrin has been purified from the urine of preterm infants fed human milk. The maternal origin of this lactoferrin, and the integrity of its primary structure have been documented. Computer analyses of the circular dichroism spectra revealed a composite secondary structure for the urinary lactoferrin that was indistinguishable from that of purified human milk lactoferrin and similar to that observed in the crystal structure. Intact function was suggested by iron binding; an approximate 2:1 molar ratio of iron to lactoferrin was confirmed. Thus, maternal lactoferrin is absorbed intact by the preterm infant and appears to remain structurally and functionally intact within the circulatory system and during urinary excretion. It is possible, therefore, that maternal lactoferrin has an immunoregulatory influence in newborn infants fed human milk.
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Recién Nacido de Bajo Peso/fisiología , Recien Nacido Prematuro/fisiología , Lactoferrina/orina , Leche Humana , Dicroismo Circular , Femenino , Humanos , Recién Nacido de Bajo Peso/orina , Recién Nacido , Recien Nacido Prematuro/orina , Hierro/análisis , Lactoferrina/química , Unión Proteica , Estructura Secundaria de ProteínaRESUMEN
The origin of intact (78-kD) lactoferrin found in the urine of human milk-fed preterm infants was investigated using human milk containing proteins enriched with [13C]leucine and [15N2]lysine or [2H4]lysine. Mothers of infants selected for the study were infused i.v. with [13C] leucine and [15N2]lysine or [2H4]lysine to label milk proteins. The labeled milk was collected from each mother, pooled, fortified with a lyophilized human milk fraction, and fed to her preterm infant by continuous orogastric infusion for a period of 48 h. Urine was collected from each infant for 96 h. Intact lactoferrin (78 kD) and DNA-binding lactoferrin fragments (51 and 39 kD) were purified from the urine by affinity chromatography on columns of immobilized single-stranded DNA-agarose. The concentration and isotopic enrichment of the intact lactoferrin and DNA-binding fragments were determined separately after their isolation by high-performance reverse-phase (phenyl) chromatography. Mass spectral analyses indicated that the isotopic enrichment of the purified urinary lactoferrin was 87 to 100% of that in the labeled human milk lactoferrin. Similar results were obtained for the isolated DNA-binding lactoferrin fragments. The ratios of isotopically labeled leucine to lysine in the purified milk lactoferrins and urinary lactoferrins were similar for each mother/infant pair. Isotopically labeled lysine, added to the milk as free amino acid, was not incorporated into the purified urinary lactoferrin. These results demonstrate that undegraded (78-kD) lactoferrin of maternal origin is absorbed by the gut and excreted intact in the urine of preterm infants; nearly all of the urinary lactoferrin was of maternal origin. The possible immunoregulatory functions of the absorbed intact, maternal lactoferrin are discussed.
Asunto(s)
Proteínas de Unión al ADN/orina , Lactoferrina/orina , Leche Humana/metabolismo , Fragmentos de Péptidos/orina , Sistema Digestivo/metabolismo , Humanos , Recién Nacido , Recien Nacido Prematuro , Isótopos , Leucina/metabolismo , Lisina/metabolismoRESUMEN
The molecular forms of lactoferrin (LF) were examined in stools and urine collected at 2.5 or 5 wk of age from very low birth wt infants fed either a cow's milk formula or a fortified human milk preparation. LF was not found by Western blotting in excreta from infants fed cow's milk. In contrast, intact and fragmented forms of LF were detected in stools and concentrated urine of each infant who received human milk. Only intact LF was detected in the fortified human milk preparation, whereas many types of LF fragments were present in the stools and urine. The approximate molecular wt of the most prominent fragments were 44, 38, 34, and 32 kD. However, the stools also displayed lower molecular wt fragments that were not found in urines of those infants. The LF fragments in those excreta were similar in size to those produced in vitro by limited digestion of apo-LF with trypsin. Furthermore, fragments produced by in vitro proteolysis were immunoreactive in an ELISA for LF. Thus, the fragments of LF in stools of very low birth wt infants fed human milk appeared to be produced by in vivo proteolysis, and the close resemblance between the LF fragments in the stools and urine suggests that the urinary LF fragments originated in the gastrointestinal tract. It remains unclear, however, whether the whole LF molecules that were fragmented were derived solely from ingested LF in human milk or in part from LF produced by the infant in response to human milk feedings.
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Heces/análisis , Fenómenos Fisiológicos Nutricionales del Lactante , Recién Nacido de Bajo Peso/metabolismo , Lactoferrina/análisis , Lactoglobulinas/análisis , Leche Humana , Animales , Western Blotting , Alimentos Fortificados , Humanos , Alimentos Infantiles , Recién Nacido de Bajo Peso/orina , Recién Nacido , Lactoferrina/orina , Leche , Fragmentos de Péptidos/análisis , Fragmentos de Péptidos/orinaRESUMEN
Little is known about the metabolic fate of ingested lactoferrin in human-milk-fed term or preterm infants. Enzyme-linked immunosorbent assays and sodium dodecyl sulfate/polyacrylamide gradient gel electrophoresis (SDS-PAGGE) with immunoblots have demonstrated that the urinary excretion of lactoferrin by preterm infants fed exclusively human milk exceeded that by formula-fed infants. The origin and molecular integrity of the excreted lactoferrin, however, are unclear. We have developed extraction and separation procedures involving a stationary phase of immobilized single-stranded DNA (ssDNA) that allows the efficient (greater than 80%) and rapid isolation of pure, intact lactoferrin from infants' urine. We purified lactoferrin to apparent homogeneity from infants' urine in one step, using the immobilized ssDNA with mobile phases containing up to 6 mol of urea per liter. The purified lactoferrin was evaluated by SDS-PAGGE; silver-staining revealed one protein band at 78 kDa; immunoblots confirmed the presence of intact lactoferrin. High-performance affinity chromatography with use of immobilized metal ion (Cu2+) suggested the presence of intact, iron-saturated lactoferrin. Subsequent chromatography on high-performance reversed-phase (C18) columns independently verified sample identity and purity.
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Lactancia Materna , Recien Nacido Prematuro , Lactoferrina/orina , Lactoglobulinas/orina , Western Blotting , Cromatografía de Afinidad , Cromatografía Líquida de Alta Presión , ADN , Ensayo de Inmunoadsorción Enzimática , Humanos , Recién Nacido , Leche Humana , SefarosaRESUMEN
The effects of fortified human milk feedings on the urinary excretion of lactoferrin, lysozyme, secretory component, IgA, and secretory IgA antibodies to Escherichia coli O antigens were investigated in very low birth wt infants. Infants were maintained on either a human milk or a cow's milk preparation. The amounts of each immune factor that were ingested and excreted were quantified during balance studies conducted at 2.5 and 5 wk of age. Serum levels of these immune factors were similar in both feeding groups. The urinary excretion of all factors except lysozyme was 7- to 150-fold greater in infants fed human milk than in those fed cow's milk formula. IgA was the only factor for which the amount of the factor excreted correlated with the amount ingested. Fragments as well as whole molecules of lactoferrin were found in the urine of the infants fed human milk, but the molecular sizes of the excreted proteins exceeded those normally filtered by the kidneys. Therefore, the genesis of the enhanced levels of host defense factors in the urine of infants fed human milk is not clear. Gastrointestinal absorption and subsequent renal excretion as well as enhanced production of immune factors in the infant's urinary tract are possible explanations.
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Inmunoglobulina A Secretora/orina , Fenómenos Fisiológicos Nutricionales del Lactante , Recién Nacido de Bajo Peso/orina , Lactoferrina/orina , Lactoglobulinas/orina , Leche Humana , Muramidasa/orina , Humanos , Recién Nacido de Bajo Peso/inmunología , Recién NacidoRESUMEN
To investigate the influence of breast feeding on mucosal immunity the concentrations and daily outputs of IgA and lactoferrin in urine were measured in 10 breast fed and 12 infants fed on formula milk at 6 and 12 weeks of age. The concentrations and outputs of secretory IgA in urine were significantly higher in the breast fed group by a factor of three. The secretion of IgA in urine by the breast fed infants was characteristic of the baby and was not related to the intake of IgA from breast milk. Lactoferrin concentrations were similar in the two groups at both ages. In addition to secretory IgA, two thirds of all samples contained proteins with alpha chain but no secretory component antigenic determinants. Breast feeding seems to increase the local production of secretory IgA into the urinary tract during early childhood, thus providing enhanced protection from infection.
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Lactancia Materna , Inmunoglobulina A/orina , Humanos , Inmunoglobulina A Secretora/orina , Lactante , Alimentos Infantiles , Lactoferrina/orina , Leche Humana/inmunologíaRESUMEN
This study concerns the sexual functions of 101 patients who had undergone bilateral retroperitoneal lymph node dissection for stage I or II nonseminomatous testicular cancer between 1969 and 1982. All patients were without evidence of disease after at least 4 years of follow-up. Antegrade ejaculation was present in 12 patients, while 89 patients experienced "dry ejaculation." Urine collected after intercourse or masturbation from 75 patients with dry ejaculation showed retrograde ejaculation in 55 and lack of ejaculatory emission into the urethra in 20 patients. Regarding other sexual functions, 17 patients had a diminished sexual desire (especially those patients who had received radiotherapy), 12 experienced difficulty reaching organism, and 6 complained of erectile dysfunction. The incidence of a contralateral hydrocele developing after retroperitoneal lymph node dissection seems to correlate with ligation of the contralateral spermatic vessels and their lymphatics. A review of the literature is presented comparing the types of dissection with the incidence of sexual disorders after retroperitoneal lymph node dissection. Since preserving normal ejaculation and fertility is important, a modified or unilateral retroperitoneal lymph node dissection, when required, is advocated. In patients, with stage I disease the therapy may be limited to an orchiectomy without lymph node dissection. In patients with retroperitoneal lymph node metastases combination chemotherapy with cisplatin and tumor excision gives good results. Patients with true retrograde ejaculation can be treated with alpha-sympathomimetic drugs such as imipramine HCl, and thus be offered the chance of fatherhood by coitus.
Asunto(s)
Escisión del Ganglio Linfático/efectos adversos , Disfunciones Sexuales Fisiológicas/etiología , Neoplasias Testiculares/cirugía , Adulto , Antígenos/orina , Eyaculación , Humanos , Lactoferrina/orina , Masculino , Persona de Mediana Edad , Enfermedades del Pene/etiología , Erección Peniana , Espacio Retroperitoneal , Espermatozoides/inmunología , Espermatozoides/patología , Neoplasias Testiculares/radioterapia , Neoplasias Testiculares/orina , Orina/citologíaRESUMEN
Vesicoureteral reflux was defined as unilateral in 17 of 36 cases. Urine was collected from both kidneys. Urine specific gravity and osmolality were significantly reduced; beta 2-microglobulin was elevated and lactoferrin depressed on the reflux side. We also observed a significant correlation between specific gravity or osmolality on one side and lactoferrin on the other side, and between osmolality and beta 2-microglobulin. These observations showed a depressed concentrating capacity and reduced tubular absorption of beta 2-microglobulin suggesting reduced tubular function on the reflux side.