RESUMEN
Objectives The objective of this study was to test the correlation of urinary podocyte number (U-Pod) and urinary podocalyxin levels (U-PCX) with histology of lupus nephritis. Methods This was an observational, cross-sectional study. Sixty-four patients were enrolled: 40 with lupus nephritis and 24 without lupus nephritis (12 lupus nephritis patients in complete remission and 12 systemic lupus erythematosus patients without lupus nephritis). Urine samples were collected before initiating treatment. U-Pod was determined by counting podocalyxin-positive cells, and U-PCX was measured by sandwich ELISA, normalized to urinary creatinine levels (U-Pod/Cr, U-PCX/Cr). Results Lupus nephritis patients showed significantly higher U-Pod/Cr and U-PCX/Cr compared with patients without lupus nephritis. U-Pod/Cr was high in proliferative lupus nephritis (class III±V/IV±V), especially in pure class IV (4.57 (2.02-16.75)), but low in pure class V (0.30 (0.00-0.71)). U-Pod/Cr showed a positive correlation with activity index ( r=0.50, P=0.0012) and was independently associated with cellular crescent formation. In contrast, U-PCX/Cr was high in both proliferative and membranous lupus nephritis. Receiver operating characteristic analysis revealed significant correlation of U-Pod/Cr with pure class IV, class IV±V and cellular crescent formation, and the combined values of U-Pod/Cr and U-PCX/Cr were shown to be associated with pure class V. Conclusions U-Pod/Cr and U-PCX/Cr correlate with histological features of lupus nephritis.
Asunto(s)
Nefritis Lúpica/patología , Nefritis Lúpica/orina , Podocitos/patología , Sialoglicoproteínas/orina , Adulto , Biomarcadores/orina , Estudios de Casos y Controles , Creatinina/orina , Estudios Transversales , Femenino , Humanos , Japón , Modelos Lineales , Lupus Eritematoso Sistémico/complicaciones , Masculino , Persona de Mediana Edad , Curva ROCRESUMEN
BACKGROUND: Acquired skin hypopigmentation has many etiologies, including autoimmune melanocyte destruction, skin aging, inflammation, and chemical exposure. Distinguishing lesions from normally pigmented skin is clinically important to precisely assess disease severity. However, no gold standard assessment method has been reported. We aimed to investigate whether spectrophotometers are useful for assessing vitiligo and rhododendrol (4-(4-hydroxyphenol)-2-butanol) (Rhododenol® )-induced leukoderma disease severity by quantifying skin color. METHODS: Mexameter® MX18 and CM-700d spectrophotometer were used for assessing vitiligo/leukoderma by measuring melanin index, L*a*b* color space, and ΔE*ab value, which represents the color difference between two subjects and is calculated by the values of L*a*b*. RESULTS: MX18 and CM-700d can quantitatively distinguish vitiligo/leukoderma from normally pigmented skin based on melanin index. CM-700d consistently quantified the color of vitiligo/leukoderma lesions and surrounding normally pigmented skin in L*a*b* color spaces and ΔE*ab. ΔE*ab is well correlated with melanin index and clinical appearance. CONCLUSION: ΔE*ab has been frequently used in aesthetic dentistry; however, current study is the first to use it in the measurement of skin color. ΔE*ab seems to be a useful parameter to evaluate the color contrast between vitiligo/leukoderma and surrounding normally pigmented skin and can be used to evaluate disease severity and patient's quality of life.
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Hipopigmentación/inducido químicamente , Pigmentación de la Piel/fisiología , Vitíligo/patología , Adulto , Femenino , Humanos , Hipopigmentación/patología , Masculino , Melaninas/metabolismo , Persona de Mediana Edad , EspectrofotometríaRESUMEN
BACKGROUND: Although immunotherapy is thought to be a promising cancer treatment, most patients do not respond to immunotherapy. In this post hoc analysis of a phase 1/2 study, associations of programmed death ligand 1 (PD-L1), PD-L2, and HLA class I expressions with responses to dendritic cells (DCs)-based immunotherapy were investigated in patients with advanced sarcoma. METHODS: This study enrolled 35 patients with metastatic and/or recurrent sarcomas who underwent DC-based immunotherapy. The associations of PD-L1, PD-L2, and HLA class I expressions in tumor specimens, which were resected before immunotherapy, with immune responses (increases of IFN-γ and IL-12) and oncological outcomes were evaluated. RESULTS: Patients who were PD-L2 (+) showed lower increases of IFN-γ and IL-12 after DC-based immunotherapy than patients who were PD-L2 (-). The disease control (partial response or stable disease) rates of patients who were PD-L1 (+) and PD-L1 (-) were 0% and 22%, respectively. Disease control rates of patients who were PD-L2 (+) and PD-L2 (-) were 13% and 22%, respectively. Patients who were PD-L1 (+) tumors had significantly poorer overall survival compared with patients who were PD-L1 (-). No associations of HLA class I expression with the immune response or oncological outcomes were observed. CONCLUSIONS: This study suggests that PD-L1 and PD-L2 are promising biomarkers of DC-based immunotherapy, and that addition of immune checkpoint inhibitors to DC-based immunotherapy may improve the outcomes of DC-based immunotherapy.
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Antígeno B7-H1/metabolismo , Antígenos de Histocompatibilidad Clase I/metabolismo , Inmunoterapia , Proteína 2 Ligando de Muerte Celular Programada 1/metabolismo , Sarcoma/terapia , Adulto , Biomarcadores de Tumor/metabolismo , Células Dendríticas , Femenino , Humanos , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Masculino , Sarcoma/inmunología , Sarcoma/mortalidad , Sarcoma/patología , Resultado del TratamientoRESUMEN
OBJECTIVES: To explore the association between dental erosion and gastro-oesophageal reflux disease (GORD), we used an animal model of GORD. MATERIALS AND METHODS: We performed an operation to force gastro-duodenal contents reflux in male Wistar rats, and examined the teeth in the reflux rats at 15 or 30 weeks postoperatively. Dental erosion was evaluated based on a slightly modified index from a previous report. Estimation of pH was employed in the oesophageal and gastric contents. RESULTS: Macroscopically, dental erosion was only detected in the reflux rats. Histopathologically, dentin exposure was detected in three of the seven cases after 30 weeks. Alveolar bone destruction and osteomyelitis were also noted in severe cases. The pH of the oesophageal and stomach contents was 6.93 +/- 0.15 and 3.7 +/- 0.39, respectively. CONCLUSIONS: We confirmed the relationship between dental erosion and GORD. First step of dental erosion caused by GORD is the loss of surface enamel induced by regurgitation of an acidic liquid and acidic gas. Subsequently, further destruction of dental hard tissues and tooth supporting structure is accelerated by mixed juice with gastric and duodenal contents. The reflux animal model is a useful tool to examine the mechanism of dental erosion in GORD.
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Modelos Animales de Enfermedad , Reflujo Gastroesofágico/complicaciones , Erosión de los Dientes/etiología , Pérdida de Hueso Alveolar/etiología , Anastomosis Quirúrgica , Animales , Esmalte Dental/patología , Dentina/patología , Esófago/fisiopatología , Esófago/cirugía , Reflujo Gastroesofágico/fisiopatología , Contenido Digestivo/química , Concentración de Iones de Hidrógeno , Yeyuno/cirugía , Masculino , Enfermedades Mandibulares/etiología , Diente Molar/patología , Osteomielitis/etiología , Ratas , Ratas Wistar , Factores de Tiempo , Erosión de los Dientes/clasificaciónRESUMEN
We report a rare case of choledochal cyst (CC) associated with congenital duodenal atresia (DA) and annular pancreas (AP). A girl was born at 37 weeks of gestation weighing 2,974 g with a prenatal diagnosis of DA. She underwent a duodenoduodenostomy for type III DA with an AP 1 day after birth. At 4 years of age, she was admitted for evaluation of cholangitis and pancreatitis. Radiological studies demonstrated a fusiform-type CC with pancreaticobiliary maljunction (PBMJ). Excision of the CC and hepaticojejunostomy were performed. The patient was discharged without complications. Despite the fact that CC, DA, and AP are embryologically closely related entities, to the best of our knowledge, only eight such cases have been documented. We must be aware of the possible combination of CC in the follow-up of the patients with DA associated with AP.
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Quiste del Colédoco/complicaciones , Duodeno/anomalías , Atresia Intestinal/complicaciones , Preescolar , Quiste del Colédoco/diagnóstico , Femenino , Humanos , Atresia Intestinal/diagnósticoRESUMEN
Bentall operation has been a gold-standard procedure for the aortic root reconstruction in the patients with aortic root aneurysms and aortic valve insufficiency (AI). Recently, native aortic valve preserving operation has been widely spread. In this study, we described the results of aortic root reconstruction in our institution and evaluated our operative methods. We selected Bentall operation for severely damaged or cardiac redo cases, and David operation for simple annuloaortic ectasia and AI. Recently, we have utilized Valsalva graft which provides the advantage of capability of larger prosthetic valve implantation and better hemodynamic performance.
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Aorta/cirugía , Procedimientos Quirúrgicos Cardiovasculares/métodos , Adulto , Anciano , Anciano de 80 o más Años , Válvula Aórtica , Prótesis Vascular , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
OBJECTIVES: To determine the association between sarcopenia and overactive bladder (OAB) in elderly diabetic patients using the Japanese version of SARC-F called SARC-F-J. DESIGN: Cross-sectional study. SETTINGS AND PARTICIPANTS: The study included 329 elderly diabetic patients (aged ≥65 years) who regularly visited the outpatient clinic at Community hospital in Japan. MEASUREMENTS: The condition of OAB was evaluated using the OAM symptom score, which involves a self-administered questionnaire, and sarcopenia was evaluated using the self-administered SARC-F-J questionnaire comprising five items. The odds ratio for OAB due to sarcopenia was calculated using multiple logistic regression analysis, with OAB as the dependent variable and sarcopenia as the explanatory variable. RESULTS: A total of 329 patients (186 males, 143 females) were included for analysis in the present study. Of these patients, 22.9% had sarcopenia and 18.7% had OAB. After adjusting the variables, the odds ratio for OAB due to sarcopenia was 4.46 (95% confidence interval [CI], 1.14-17.36, P = 0.031) and 2.09 (95% CI, 0.52-8.26, P = 0.293) for males and females, respectively. CONCLUSION: This study found that sarcopenia was significantly associated with OAB in elderly diabetic male patients based on SARC-F-J. Moreover, the possibility of the development of OAB should be considered during the medical examinations of elderly diabetic male patients with sarcopenia.
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Complicaciones de la Diabetes/complicaciones , Sarcopenia/complicaciones , Vejiga Urinaria Hiperactiva/etiología , Anciano , Anciano de 80 o más Años , Estudios Transversales , Femenino , Humanos , Masculino , Encuestas y CuestionariosRESUMEN
Platelet-activating factor (PAF) acetylhydrolase has been recognized as an enzyme that inactivates PAF. We developed a convenient and reproducible method for determining human serum PAF acetylhydrolase activity. The assay was based on measurement of [14C]acetate produced from 1-O-alkyl-2-[14C]-acetyl-sn-glycero-3-phosphocholine upon precipitation of the complex of radioactive substrate and albumin with TCA. The apparent Km value of PAF acetylhydrolase (near the physiological concentration of serum protein) was 1.5 X 10(-4) M PAF. 32 subjects with serum PAF acetylhydrolase deficiency were found among 816 healthy Japanese adults. The low PAF acetylhydrolase activity in the deficient serum might not be due to the presence of enzyme inhibitor. Both the sensitivity to PAF and the metabolism of PAF in platelets from PAF acetylhydrolase-deficient subjects were almost the same as those of normal subjects. Deficiency in serum PAF acetylhydrolase appeared to be transmitted by autosomal recessive heredity among five Japanese families. Among healthy adults, healthy children, and asthmatic children, who were grouped into five classes on the basis of respiratory symptoms (remission, wheezy, mild, moderate, and severe groups), the probability of PAF acetylhydrolase deficiency was significantly higher in groups with severe symptoms (moderate and severe) (P less than 0.01). These results suggest that deficiency of serum PAF acetylhydrolase might be one of the factors leading to severe respiratory symptoms in asthmatic children.
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Asma/enzimología , Fosfolipasas A/sangre , Fosfolipasas/sangre , 1-Alquil-2-acetilglicerofosfocolina Esterasa , Adolescente , Adulto , Asma/genética , Niño , Colesterol/sangre , Familia , Humanos , Lipoproteínas LDL/sangre , Métodos , Linaje , Fosfolipasas A/deficiencia , Agregación PlaquetariaRESUMEN
Exposure of platelets to shear stress leads to aggregation in the absence of exogenous agonists. We have now found that different adhesive proteins and platelet membrane glycoproteins are involved in aggregation depending on the shear stress conditions and the concentration of divalent cations in the medium. When blood is collected with trisodium citrate as anticoagulant, which causes a decrease in the levels of external ionized calcium ([Ca2+]o), platelet aggregation can be induced under low shear force (12 dyn/cm2) and is mediated by fibrinogen binding to the glycoprotein IIb-IIIa complex. Aggregates formed under these conditions are not stable, and when shear force is increased to 68 dyn/cm2, disaggregation results. By contrast, platelets from blood collected with hirudin as anticoagulant, wherein [Ca2+]o is within normal plasma levels, do not undergo low shear-induced aggregation; however, after exposure to a shear force above 80 dyn/cm2, aggregation is observed but only when von Willebrand factor is present and can interact with both its platelet binding sites, glycoprotein Ib-IX and glycoprotein IIb-IIIa. Fibrinogen is not involved in high shear-induced aggregation which, in fact, occurs normally in patients with severe afibrinogenemia. Thus, von Willebrand factor in the absence of exogenous agonists can mediate platelet aggregation in experimental conditions that may mimic the hemorheological situation of partially occluded arteries. This pathway of platelet aggregation involving only one adhesive ligand and two membrane adhesion receptors may play a relevant role in thrombogenesis.
Asunto(s)
Fibrinógeno/metabolismo , Agregación Plaquetaria , Glicoproteínas de Membrana Plaquetaria/metabolismo , Factor de von Willebrand/metabolismo , Afibrinogenemia/fisiopatología , Síndrome de Bernard-Soulier/fisiopatología , Humanos , Técnicas In Vitro , Unión Proteica , Estrés Mecánico , Trombastenia/fisiopatología , Factores de Tiempo , Enfermedades de von Willebrand/fisiopatologíaRESUMEN
This article shows a prospective study investigating bowel function after transanal rectal resection with internal and external sphincterectomy for low rectal cancer. Eight patients underwent standard low anterior resection with colonic J-pouch anal anastomosis (LARJ), and eight patients underwent transanal rectal resection with internal and external sphincter resection (IESR). Manometry, manovolumetry, transit time study, and a questionnaire were performed before and after the operation. Six and 12 months after the operation, maximum resting pressure and squeezing pressure were significantly lower in IESR group than in LARJ group, whereas there was no significant difference between the two groups in terms of constant sensation, maximum tolerable volume, or neorectal compliance. Although the functional score of the IESR group remained low at 6 months after the operation in comparison with the LARJ group, it improved at 12 months after the operation. Transanal rectal resection with internal and external sphincterectomy showed usefulness in preserving bowel function and avoiding permanent colostomy.
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Canal Anal/cirugía , Neoplasias del Recto/cirugía , Anciano , Anciano de 80 o más Años , Canal Anal/fisiopatología , Anastomosis Quirúrgica/métodos , Neoplasias del Ano/cirugía , Femenino , Estudios de Seguimiento , Tránsito Gastrointestinal/fisiología , Humanos , Masculino , Manometría , Persona de Mediana Edad , Estudios Prospectivos , Neoplasias del Recto/fisiopatología , Estadísticas no Paramétricas , Resultado del TratamientoRESUMEN
It is intrinsically difficult to directly measure a specific protein reduction that is mediated by nonselective autophagy, because the degradation proceeds at a relatively slow pace, and the residual nondegraded part becomes the background. Several methods for measuring nonselective autophagy have been reported in the past. One classical simple method is called bulk degradation assay, which measures the release of degraded amino acids after radioisotope labeling of the total cellular proteins. In 1995, we developed a quantitative Pho8â³60 assay in the yeast, Saccharomyces cerevisiae, for studying autophagy, which is widely used for its advantages that are described in the following sections. Another method used in recent times is the GFP-based processing assay in yeast. We will describe these two methods in this chapter.
Asunto(s)
Fosfatasa Alcalina/metabolismo , Familia de las Proteínas 8 Relacionadas con la Autofagia/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/citología , Fosfatasa Alcalina/genética , Autofagia , Familia de las Proteínas 8 Relacionadas con la Autofagia/genética , Pruebas de Enzimas/métodos , Expresión Génica , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Vacuolas/genética , Vacuolas/metabolismoRESUMEN
Long-term insulin secretion was investigated in seven biohybrid capillary devices seeded with canine islets. Approximately 50,000 islets could be isolated from a single canine pancreas using collagenase digestion in conjunction with the recently described Velcro technique. Devices were perfused with tissue culture medium 199 containing 300 mg/dl glucose. Insulin secretion fell during the first 1-2 days of culture from approximately 18 to 6 U/day. After 3-4 wk of perfusion, however, there was a gradual rise in insulin output that reached greater than 15 U/day after 7 wk. Insulin output eventually stabilized at 18-20 U/day. Two of these devices were studied for 80 and 86 days, respectively, and continued to secrete these same amounts of insulin. Glucose-induced insulin release was studied after 1 and 4 wk of culture and was well preserved.
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Sistemas de Infusión de Insulina , Insulina/metabolismo , Islotes Pancreáticos/metabolismo , Animales , Bovinos , Células Cultivadas , Medios de Cultivo , Perros , Femenino , Glucosa/metabolismo , Haplorrinos , Humanos , Secreción de Insulina , Islotes Pancreáticos/citología , Masculino , Ratas , Factores de TiempoRESUMEN
Patients with open bite often show a weak occlusal force and temporomandibular disorders (TMDs). If these are the main cause of open bite, it may be hypothesized that both pre-pubertal and adult open-bite patients would show a weak occlusal force and abnormal condylar motion. The purpose of this study was to test this hypothesis. Test group subjects consisted of 13 consecutive pre-pubertal and 13 adult patients with anterior open bite. They were compared with age-matched normal subjects. The adult open-bite group showed a weaker occlusal force and a shorter range of condylar motion compared with the control subjects. In the pre-pubertal subjects, however, there were no significant differences in the occlusal force and range of condylar motion between the open-bite and control groups. Therefore, these results suggest that a weak occlusal force or TMDs may not be the main cause of open bite.
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Fuerza de la Mordida , Mordida Abierta/fisiopatología , Rango del Movimiento Articular/fisiología , Trastornos de la Articulación Temporomandibular/fisiopatología , Articulación Temporomandibular/fisiología , Adolescente , Adulto , Cefalometría , Niño , Femenino , Humanos , Masculino , Cóndilo Mandibular/fisiología , Análisis por Apareamiento , Mordida Abierta/complicaciones , Articulación Temporomandibular/fisiopatología , Trastornos de la Articulación Temporomandibular/etiologíaRESUMEN
Leukocyte extravasation is essential for subsequent inflammation and the immune response. Extravasation can be divided into at least three steps; rolling, firm adhesion, and transendothelial migration. Although the mechanisms involved in the first two steps have been fairly well documented, the last step is complex and largely remains to be clarified. This review focuses on the possible role of GPI-anchored proteins on leukocytes in the regulation of their transendothelial migration. In addition to regulation by urokinase and its receptor, which has been regarded as the main modulator, we draw attention to a novel GPI-anchored protein (GPI-80) on human phagocytes that may be involved in regulating leukocyte adhesion and migration. The high degree of homology of GPI-80 with vanin-1, which is expressed on vascular tissues and is involved in prethymic cell homing into the thymus, raises the possibility that there is a family of molecules, including GPI-80 and vanin-1, that may be involved in leukocyte transendothelial migration. The possible role of soluble GPI-anchored proteins in this process is also discussed.
Asunto(s)
Adhesión Celular/fisiología , Quimiotaxis de Leucocito/fisiología , Glicosilfosfatidilinositoles/fisiología , Leucocitos/citología , Amidohidrolasas/química , Secuencia de Aminoácidos , Biotinidasa , Moléculas de Adhesión Celular/química , Moléculas de Adhesión Celular/fisiología , Endotelio Vascular/citología , Proteínas Ligadas a GPI , Humanos , Hidrolasas , Integrinas/fisiología , Leucocitos/química , Leucocitos/metabolismo , Ligandos , Modelos Biológicos , Datos de Secuencia Molecular , Especificidad de Órganos , Fagocitos/citología , Fagocitos/metabolismo , Inhibidor 1 de Activador Plasminogénico/fisiología , Receptores de Superficie Celular/fisiología , Receptores del Activador de Plasminógeno Tipo Uroquinasa , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Timo/citología , Activador de Plasminógeno de Tipo Uroquinasa/fisiologíaRESUMEN
We previously reported a novel glycosylphosphatidylinositol (GPI)-anchored glycoprotein (tentatively designated GPI-80) on human leukocytes that may be involved in the regulation of neutrophil adherence and migration. In this study, we examined by immuno- and scanning electron microscopy, the distribution of GPI-80 on neutrophil surfaces. GPI-80 was diffusely distributed on the surface of resting neutrophils and on the peripheral areas of adherent cells after stimulation with N-formyl-methionyl-leucyl-phenylalanine. After longer stimulation (60 min), GPI-80 decreased in number and was again diffusely distributed on the surfaces of round neutrophils. Few GPI-80 were detected on the ventral surfaces of adherent neutrophils. Clusters of GPI-80 were detected on the forward surfaces of neutrophils transmigrating through pores of nitrocellulose membranes. These results may give a morphological background of possible role of GPI-80 for neutrophil extravasation.
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Moléculas de Adhesión Celular/metabolismo , Movimiento Celular/fisiología , Glicosilfosfatidilinositoles/metabolismo , Neutrófilos/metabolismo , Amidohidrolasas , Adhesión Celular/fisiología , Membrana Celular/metabolismo , Membrana Celular/ultraestructura , Proteínas Ligadas a GPI , Humanos , Hidrolasas , Microscopía Electrónica de Rastreo , Microscopía Inmunoelectrónica , N-Formilmetionina Leucil-Fenilalanina/farmacología , Neutrófilos/citología , Neutrófilos/ultraestructura , Estimulación QuímicaRESUMEN
This report describes a renal dysplastic lesion associated with renal agenesis in a 3-year-old dog with chronic renal failure. Haematological examination revealed non-regenerative anaemia, azotaemia, increased creatinine and hyperphosphataemia. At necropsy, the right kidney and right ureter could not be identified. The left kidney was slightly enlarged, with a reduced cortico-medullary ratio. Histologically, the medulla of the left kidney had persistent mesenchyme and primitive tubules (tall pseudostratified columnar epithelium), dilated collecting ducts lined by flattened epithelium, and adenomatoid proliferation of cuboidal epithelium; fetal or immature glomeruli could not be identified. To our knowledge, this is the first report of a renal dysplastic lesion with unilateral agenesis in animals.
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Enfermedades de los Perros/congénito , Fallo Renal Crónico/patología , Fallo Renal Crónico/veterinaria , Riñón/anomalías , Riñón/patología , Animales , Enfermedades de los Perros/patología , Perros/anomalías , Fallo Renal Crónico/complicaciones , Masculino , Uréter/anomalías , Uréter/patologíaRESUMEN
Tissue-specific and developmental stage-dependent expression of a novel Dopa/tyrosine sulfotransferase in Sprague-Dawley rats was examined. Both immunoblot and Northern blot analyses showed that the enzyme was expressed predominantly in liver and to a lesser extent in kidney. Its expression could not be detected in nine other organs tested. Livers from different age groups of male or female rats were examined for the developmental regulation of the expression of the Dopa/tyrosine sulfotransferase. Results from immunoblot and Northern blot analyses revealed that the enzyme was present at a very low level in livers of 1-day-old to 2-week-old rats, and gradually increased to a maximum level in rats older than 2 months. Data from the enzymatic assays also showed a similar trend of expression in both male and female rats. The Dopa/tyrosine sulfotransferase activities detected in liver samples of the 8-week-old male and female rats were, respectively, 8.6 and 6.6 times that of the activities detected in liver samples of the 1-day-old male and female rats. These data provide a foundation for the future investigation of the cis- and trans-acting factors involved in the regulation of the tissue-specific and developmental stage-dependent expression of this enzyme.
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Envejecimiento/metabolismo , Sulfotransferasas/biosíntesis , Animales , Densitometría , Femenino , Riñón/enzimología , Hígado/enzimología , Masculino , Ratas , Ratas Sprague-Dawley , Especificidad por SustratoRESUMEN
A novel enzyme which specifically hydrolyzes N-acetylglucosaminyl pyrophosphorylpolyprenol to liberate N-acetylglucosamine was found in membranes of Bacillus polymyxa AHU 1385. The enzyme seems to be inactive toward alpha-N-acetylglucosaminyl phosphorylundecaprenol, beta-N-acetylglucosaminyl phosphorylundecaprenol, N-acetylglucosamine 1-phosphate, N-acetylglucosamine 1-pyrophosphate, or UDP-N-acetylglucosamine. Much lower activities of the same enzyme were also found in membranes of several other strains of Bacilli.
Asunto(s)
Acetilglucosamina/metabolismo , Bacillus/enzimología , Proteínas Bacterianas/aislamiento & purificación , Glucosamina/análogos & derivados , Proteínas de la Membrana/aislamiento & purificación , Hidrolasas Diéster Fosfóricas/aislamiento & purificación , Monosacáridos de Poliisoprenil Fosfato/metabolismo , Azúcares de Poliisoprenil Fosfato/metabolismo , Proteínas Bacterianas/metabolismo , Hidrólisis , Proteínas de la Membrana/metabolismo , Hidrolasas Diéster Fosfóricas/metabolismo , Especificidad de la Especie , Especificidad por SustratoRESUMEN
Two hybridoma cell lines producing monoclonal antibodies WGA-1 and PL7-6, reactive only with thrombin-stimulated human platelet have been established. Both these antibodies were investigated for their specific reactivity against GMP-140, based on the amino acid composition analysis of immunopurified antigen and N terminal amino acid sequencing of its protease fragments. A two-site enzyme immunoassay for quantification of human GMP-140 was developed using WGA-1 monoclonal antibody immobilized on 96-well microplates and horseradish peroxidase-labeled PL7-6 monoclonal antibody as detector. The assay was able to measure GMP-140 in serum and plasma with a sensitivity of about 5 ng/ml and a precision better than 10%. This assay will be useful for the detection of GMP-140 derived from platelets or endothelium in biological fluids and tissue extracts.
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Anticuerpos Monoclonales/inmunología , Glicoproteínas de Membrana Plaquetaria/análisis , Secuencia de Aminoácidos , Animales , Femenino , Humanos , Técnicas para Inmunoenzimas , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Selectina-P , Glicoproteínas de Membrana Plaquetaria/inmunologíaRESUMEN
Dipeptidyl aminopeptidase IV activity staining was performed in various thyroid tissues to evaluate this enzyme activity as a thyroid tumor marker. A total of 195 thyroid tissues were tested for their enzyme activity expression. All papillary and follicular carcinomas, 40 cases and 3 cases, respectively, showed enzyme activity, although two other carcinomas, one medullary and one anaplastic, were not stained. Follicular adenoma expressed enzyme activity in 4 of 26 cases. Fifty-two cases with adenomatous goiter, 54 with Graves' disease and 13 with chronic thyroiditis were judged to be negative. Five normal thyroids expressed no activity except for occasional positive staining of capillary endothelia. These data suggest that dipeptidyl aminopeptidase IV activity staining is very useful for pathological diagnosis of thyroid tumors.