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1.
Cancer Research and Treatment ; : 646-657, 2018.
Article Dans Anglais | WPRIM | ID: wpr-715840

Résumé

PURPOSE: Early prediction of treatment outcomes represents an essential step towards increased treatment efficacy and survival in patients with hepatocellular carcinoma (HCC). In this study, we performed two-dimensional electrophoresis (2-DE) followed by protein profiling to identify biomarkers predictive of therapeutic outcomes in patients with HCC who received liver-directed therapy (LDTx) involving local radiotherapy (RT), and studied the underlying mechanisms of the identified proteins. MATERIALS AND METHODS: 2-DE analysis was conducted by pooling sera from patients with a good or poor prognosis; serum proteomic profiles of the two groups were compared and analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Identified proteins were confirmed via enzyme-linked immunosorbent assay. An invasion assay was performed after overexpression and knockdown of target protein in Huh7 cells. RESULTS: Levels of inter-alpha inhibitor H4 (ITIH4), fibrinogen gamma chain, keratin 9/1 complex, carbonic anhydrase I, and carbonmonoxyhemoglobin S were changed by more than 4-fold in response to LDTx. In particular, pre-LDTx ITIH4 expression was more than 5-fold higher in patients with a good prognosis, compared to patients with a poor prognosis. The migration ability of Huh7 cells was significantly suppressed and enhanced by ITIH4 overexpression and knockdown, respectively. The tumors of patients with HCC and a good prognosis expressed high levels of ITIH4, compared to those of patients with a poor prognosis. CONCLUSION: Taken together, ITIH4 may be a potential therapeutic target that could inhibit cancer metastasis, as well as a prognostic marker for patients with HCC who are receiving LDTx.


Sujets)
Humains , Marqueurs biologiques , Carbonic anhydrase I , Carboxyhémoglobine , Carcinome hépatocellulaire , Électrophorèse , Test ELISA , Fibrinogène , Spectrométrie de masse , Métastase tumorale , Pronostic , Radiothérapie , Résultat thérapeutique
2.
Rev. Asoc. Méd. Argent ; 130(3): 12-21, sept. 2017. ilus, tab
Article Dans Espagnol | LILACS | ID: biblio-973080

Résumé

La Hemoglobinuria Paroxística Nocturna (HPN) se caracteriza por hemólisis intravascular crónica mediada por complemento. Cuando se produce la hemolisis se libera a circulación Anhidrasa Carbónica- I (AC-I), una enzima que se halla en alta concentración en el eritrocito y por su bajo peso molecular filtra por el glomérulo. El objetivo del presente trabajo fue detectar la excreción de la AC-I en orina de pacientes con HPN por Electroforesis Bidimensional de Utilidad Clínica (2D UC), y compararla con otras causas de hemólisis, de origen renal y postrenal. Se evaluaron 8 pacientes con HPN sin tratamiento con eculizumab un inhibidor del C5 del complemento, y 5 de ellos postratamiento, 12 orinas de pacientes con nefritis lúpica y 10 orinas de pacientes con hemólisis postrenal. La AC-I puede estar presente en la orina, en los tres grupos, sin embargo la relación AC-I/Hemoglobina en la hemólisis intravascular está invertida en comparación con la hemolisis glomerular y post-renal. Los pacientes con HPN tratados con eculizumab no presentan AC-I, y sería de utilidad en el seguimiento de los pacientes tratados con el inhibidor del C5, para evidenciar posibles escapes hemolíticos.


Paroxysmal Nocturnal Hemoglobinuria (PNH) is characterized by chronic complement mediated haemolysis. In these conditions it might be expected that carbonic anhydrase-I (AC-I) would be liberated into the plasma and excreted in the urine, by its high concentration in the erythrocyte and low molecular weight. The objective of the present study was to detect the urinary excretion of AC-I from patients with PNH by wodimensional clinical utility electrophoresis (2D UC) and to compare it with other causes of renal and post-renal haemolysis. We evaluated 8 patients with PNH without eculizumab, a complement C5 inhibitor, 5 of them posttreatment, 12 urine of patients with lupus nephritis and 10 urine of patients with post-renal hemolysis. AC-I may be present in the urine, in all three groups, however, the AC-I/Haemoglobin ratio in intravascular haemolysis is reversed compared to glomerular and post-renal haemolysis. Patients with PNH treated with eculizumab do not have AC-I and would be useful in monitoring patients treated with the C5 inhibitor to evidence possible haemolytic leaks.


Sujets)
Humains , Hémoglobinurie paroxystique/urine , Carbonic anhydrase I/métabolisme , Carbonic anhydrase I/urine , Hémolyse , Hémoglobinurie paroxystique/traitement médicamenteux , Électrophorèse/méthodes , Examen des urines/méthodes , Lupus érythémateux disséminé/urine , Hématurie/urine , Anticorps monoclonaux humanisés/usage thérapeutique
3.
Braz. j. med. biol. res ; 48(11): 973-982, Nov. 2015. tab, graf
Article Dans Anglais | LILACS | ID: lil-762908

Résumé

Bipolar disorder (BD) is a common psychiatric mood disorder affecting more than 1-2% of the general population of different European countries. Unfortunately, there is no objective laboratory-based test to aid BD diagnosis or monitor its progression, and little is known about the molecular basis of BD. Here, we performed a comparative proteomic study to identify differentially expressed plasma proteins in various BD mood states (depressed BD, manic BD, and euthymic BD) relative to healthy controls. A total of 10 euthymic BD, 20 depressed BD, 15 manic BD, and 20 demographically matched healthy control subjects were recruited. Seven high-abundance proteins were immunodepleted in plasma samples from the 4 experimental groups, which were then subjected to proteome-wide expression profiling by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight tandem mass spectrometry. Proteomic results were validated by immunoblotting and bioinformatically analyzed using MetaCore. From a total of 32 proteins identified with 1.5-fold changes in expression compared with healthy controls, 16 proteins were perturbed in BD independent of mood state, while 16 proteins were specifically associated with particular BD mood states. Two mood-independent differential proteins, apolipoprotein (Apo) A1 and Apo L1, suggest that BD pathophysiology may be associated with early perturbations in lipid metabolism. Moreover, down-regulation of one mood-dependent protein, carbonic anhydrase 1 (CA-1), suggests it may be involved in the pathophysiology of depressive episodes in BD. Thus, BD pathophysiology may be associated with early perturbations in lipid metabolism that are independent of mood state, while CA-1 may be involved in the pathophysiology of depressive episodes.


Sujets)
Adolescent , Adulte , Femelle , Humains , Mâle , Jeune adulte , Apolipoprotéine A-I/sang , Apolipoprotéines/sang , Trouble bipolaire/sang , Carbonic anhydrase I/sang , Troubles du métabolisme lipidique/métabolisme , Lipoprotéines HDL/sang , Protéomique , Trouble bipolaire/complications , Trouble bipolaire/diagnostic , Bases de données de protéines , Diagnostic différentiel , Évolution de la maladie , Régulation négative , Trouble dépressif majeur/diagnostic , Électrophorèse bidimensionnelle sur gel , Immunotransfert , Immunoprécipitation , Troubles du métabolisme lipidique/complications , Spectrométrie de masse/méthodes
4.
Korean Journal of Anatomy ; : 215-225, 2006.
Article Dans Coréen | WPRIM | ID: wpr-644542

Résumé

The marked hemodynamic and hormonal changes of normal pregnancy are associated with striking alterations in renal physiology involving structure, dynamics, tubular function, and volume homeostasis. A number of acid-base or electrolyte disorders are associated with decreased or increased HCO3-reabsorption in the renal tubules. The present study was to examine the alterations of expression and distribution of Na+/HCO3-cotransporter (NBC), Na+/H+ exchanger-3 (NHE-3), and carbonic anhydrase I and II (CA I, II) proteins in the kidneys of non-pregnant (NP) and pregnant rats using Western blot analysis and immunohistochemistry. Sprague-Dawley female rats were studied on days 10 (P 10), 12 (P 12), 14 (P 14), 17 (P 17), and 19 (P 19) of pregnancy. Western blot analysis demonstrated that the expression of NBC, ~110 kDa at molecular mass, was increased in pregnant rats, particularly P 12, compared with NP rat. The expression of NHE-3, ~83 kDa at molecular mass, was increased in pregnant rats, particularly P 12 and P 14. The expression of CA I, ~30 kDa at molecular mass, was decreased in pregnant rats, particularly P 14, but, CA II protein, ~30 kDa at molecular mass, was similar NP rat. In immunohistochemistry, strong immunoreactivity of NBC of NP rat was exclusively detected in the basolateral membranes of S1 and S2 segment of proximal tubules whereas not in S3 segment. In pregnant rats, the pattern of cellular labeling of NBC was identical to that of NP rat, but signal intensity was increased, particularly P 12. In NHE-3, strong immunoreactivity was detected in apical membranes and brush borders of S3 segments and moderate in S1 and S2 segments. In pregnant rats, the pattern of cellular labeling was identical to that of NP rat, but the signal intensity was increased, particularly P 12 and P 14. Expression of CA I and II proteins was detected in entire collecting duct. Signal intensity was prominent in type A intercalated cells and moderate in type B intercalated cells. In pregnant rats, the pattern of cellular labeling of CA I and II proteins was identical to that of non-pregnant rat, but the signal intensity of CA I was decreased in cortical collecting duct, particularly P 14 and CA II was identical to that of NP rat. These results suggest that the regulation of NBC and NHE-3 expressions in the proximal tubules and CA I expression in cortical collecting duct may maintain HCO3-concentration during the pregnancy.


Sujets)
Animaux , Femelle , Humains , Grossesse , Rats , Hydrogénocarbonates , Technique de Western , Carbonic anhydrase I , Hémodynamique , Homéostasie , Immunohistochimie , Rein , Membranes , Microvillosités , Physiologie , Rat Sprague-Dawley , Contrôle social formel , Grèves
5.
Journal of the Korean Ophthalmological Society ; : 454-458, 2003.
Article Dans Coréen | WPRIM | ID: wpr-207755

Résumé

PURPOSE: The histiogenesis of retinoblastoma, the most common intraocular malignancy of childhood, has been investigated from the early times. But in spite of this effort, its origin has been controversial. This study was performed to investigated the cell of origin for retinoblastoma using enzyme histomchemistry for carbonic anhydrase. METHODS: We obtained enucleated eye that was diagnosed as retinoblastoma and its section was stained for hematoxylin-eosin for diagnosis of retinoblastoma. We used enzyme histomchemistry for carbonic anhydrase distinguishing Muller's cells, red-and green-sensistive cones from neuro-retinal cells. RESULTS: They were disagnosed as relatively well-differentiated retinoblastoma by hematoxylin-eosin staining and composed of tumor cells with numerous rosette. Neither numeric nor morphologic changes of Muller cells that are suspected of malignant features in enzyme histochemistry for carbonic anhydrase was found. CONCLUSIONS: The cells of retinoblastoma were originated from the two layers, inner nuclear and ganglion cell layer. The enzyme histochemistry for carbonic anhydrase is the one of the useful methods to investigate the origin of retinoblastoma although more cases is needed to assess.


Sujets)
Carbonic anhydrase I , Carbonic anhydrases , Diagnostic , Cellules épendymogliales , Pseudokystes mucoïdes juxta-articulaires , Rétinoblastome
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