RESUMO
It has been suggested that whole-genome duplication (WGD) occurred twice during the evolutionary process of vertebrates around 450 and 500 million years ago, which contributed to an increase in the genomic and phenotypic complexities of vertebrates. However, little is still known about the evolutionary process of homoeologous chromosomes after WGD because many duplicate genes have been lost. Therefore, Xenopus laevis (2n=36) and Xenopus (Silurana) tropicalis (2n=20) are good animal models for studying the process of genomic and chromosomal reorganization after WGD because X. laevis is an allotetraploid species that resulted from WGD after the interspecific hybridization of diploid species closely related to X. tropicalis. We constructed a comparative cytogenetic map of X. laevis using 60 complimentary DNA clones that covered the entire chromosomal regions of 10 pairs of X. tropicalis chromosomes. We consequently identified all nine homoeologous chromosome groups of X. laevis. Hybridization signals on two pairs of X. laevis homoeologous chromosomes were detected for 50 of 60 (83%) genes, and the genetic linkage is highly conserved between X. tropicalis and X. laevis chromosomes except for one fusion and one inversion and also between X. laevis homoeologous chromosomes except for two inversions. These results indicate that the loss of duplicated genes and inter- and/or intrachromosomal rearrangements occurred much less frequently in this lineage, suggesting that these events were not essential for diploidization of the allotetraploid genome in X. laevis after WGD.
Assuntos
Cromossomos/genética , Diploide , Evolução Molecular , Genoma , Xenopus laevis/genética , Animais , Duplicação Cromossômica , Feminino , Ligação Genética , Hibridização in Situ Fluorescente , Cariótipo , RNA Ribossômico/genética , Proteínas de Xenopus/genéticaRESUMO
The vascular complications of diabetes mellitus have been correlated with enhanced activation of protein kinase C (PKC). LY333531, a specific inhibitor of the beta isoform of PKC, was synthesized and was shown to be a competitive reversible inhibitor of PKC beta 1 and beta 2, with a half-maximal inhibitory constant of approximately 5 nM; this value was one-fiftieth of that for other PKC isoenzymes and one-thousandth of that for non-PKC kinases. When administered orally, LY333531 ameliorated the glomerular filtration rate, albumin excretion rate, and retinal circulation in diabetic rats in a dose-responsive manner, in parallel with its inhibition of PKC activities.
Assuntos
Diabetes Mellitus Experimental/complicações , Angiopatias Diabéticas/prevenção & controle , Inibidores Enzimáticos/farmacologia , Indóis/farmacologia , Isoenzimas/antagonistas & inibidores , Maleimidas/farmacologia , Proteína Quinase C/antagonistas & inibidores , Administração Oral , Albuminúria/prevenção & controle , Animais , Diabetes Mellitus Experimental/enzimologia , Diabetes Mellitus Experimental/fisiopatologia , Angiopatias Diabéticas/enzimologia , Angiopatias Diabéticas/etiologia , Diglicerídeos/metabolismo , Relação Dose-Resposta a Droga , Ativação Enzimática , Inibidores Enzimáticos/química , Taxa de Filtração Glomerular/efeitos dos fármacos , Humanos , Indóis/administração & dosagem , Indóis/química , Isoenzimas/metabolismo , Glomérulos Renais/metabolismo , Masculino , Maleimidas/administração & dosagem , Maleimidas/química , Músculo Liso Vascular/enzimologia , Fosforilação/efeitos dos fármacos , Proteína Quinase C/metabolismo , Proteína Quinase C beta , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacos , Fluxo Plasmático Renal/efeitos dos fármacos , Retina/metabolismo , Vasos Retinianos/fisiopatologia , Especificidade por SubstratoRESUMO
Increased vascular permeability and excessive neovascularization are the hallmarks of endothelial dysfunction, which can lead to diabetic macular edema and proliferative diabetic retinopathy in the eye. Vascular endothelial growth factor (VEGF) is an important mediator of ocular neovascularization and a known vasopermeability factor in nonocular tissues. In these studies, we demonstrate that intravitreal injection of VEGF rapidly activates protein kinase C (PKC) in the retina at concentrations observed clinically, inducing membrane translocation of PKC isoforms alpha, betaII, and delta and >threefold increases in retinal vasopermeability in vivo. The effect of VEGF on retinal vascular permeability appears to be mediated predominantly by the beta-isoform of PKC with >95% inhibition of VEGF-induced permeability by intravitreal or oral administration of a PKC beta-isoform-selective inhibitor that did not inhibit histamine-mediated effects. These studies represent the first direct demonstration that VEGF can increase intraocular vascular permeability through activation of PKC in vivo and suggest that oral pharmacological therapies involving PKC beta-isoform-selective inhibitors may prove efficacious for the treatment of VEGF-associated ocular disorders such as diabetic retinopathy.
Assuntos
Fatores de Crescimento Endotelial/fisiologia , Olho/irrigação sanguínea , Linfocinas/fisiologia , Proteína Quinase C/fisiologia , Retina/enzimologia , Animais , Permeabilidade Capilar , Bovinos , Células Cultivadas , Inibidores Enzimáticos/farmacologia , Indóis/farmacologia , Isoenzimas/antagonistas & inibidores , Isoenzimas/fisiologia , Maleimidas/farmacologia , Proteína Quinase C/antagonistas & inibidores , Ratos , Transdução de Sinais , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularRESUMO
OBJECTIVES: To evaluate left ventricular diastolic function and differentiate the pseudonormalized transmitral flow pattern from the normal pattern, the propagation of left ventricular early filling flow was assessed quantitatively using color M-mode Doppler echocardiography. BACKGROUND: Because the propagation of left ventricular early filling flow is disturbed in the left ventricle with impaired relaxation, quantification of such alterations should provide useful indexes for the evaluation of left ventricular diastolic function. METHODS: Study subjects were classified into three groups according to the ratio of early to late transmitral flow velocity (E/A ratio) and left ventricular ejection fraction: 29 subjects with an ejection fraction > or = 60% (control group); 34 with an ejection fraction < 60% and E/A ratio < 1 (group I); and 25 with ejection fraction < 60% and E/A ratio > or = 1 (group II). The propagation of peak early filling flow was visualized by changing the first aliasing limit of the color Doppler signals. The rate of propagation of peak early filling flow velocity was defined as the distance/time ratio between two sampling points: the point of the maximal velocity around the mitral orifice and the point in the mid-left ventricle at which the velocity decreased to 70% of its initial value. High fidelity manometer-tipped measurement was performed in 40 randomly selected subjects. RESULTS: The rate of propagation decreased in groups I and II compared with that in the control group (33.8 +/- 13.8 [mean +/- SD] and 30.0 +/- 8.6 vs. 74.3 +/- 17.4 cm/s, p < 0.001, respectively) and correlated inversely with the time constant of left ventricular isovolumetric relaxation and the minimal first derivative of left ventricular pressure (peak negative dP/dt) (r = 0.82 and r = 0.72, respectively). CONCLUSIONS: Spatial and temporal analysis of filling flow propagation by color M-mode Doppler echocardiography was free of pseudonormalization and correlated well with the invasive variables of left ventricular relaxation.
Assuntos
Diástole/fisiologia , Ecocardiografia Doppler em Cores/métodos , Disfunção Ventricular Esquerda/diagnóstico por imagem , Função Ventricular Esquerda/fisiologia , Velocidade do Fluxo Sanguíneo/fisiologia , Cateterismo Cardíaco , Estudos de Casos e Controles , Circulação Coronária/fisiologia , Estudos de Viabilidade , Humanos , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Volume Sistólico/fisiologia , Fatores de Tempo , Disfunção Ventricular Esquerda/fisiopatologiaRESUMO
Signaling triggered by polypeptide growth factors leads to the activation of their target genes. Several homeobox genes are known to be induced in response to polypeptide growth factors in early Xenopus development. In particular, Xmsx-1, an amphibian homologue of vertebrate Msx-1, is well characterized as a target gene of bone morphogenetic protein (BMP). Here, using a dominant-negative form of Xmsx-1 (VP-Xmsx-1), which is a fusion protein made with the virus-derived VP16 activation domain, we have examined whether Xmsx-1 activity is required in the endogenous ventralizing pathway. VP-Xmsx-1 induced a secondary body axis, complete with muscle and neural tissues, when overexpressed in ventral blastomeres, suggesting that Xmsx-1 activity is necessary for both mesoderm and ectoderm to be ventralized. We have also examined the epistatic relationship between Xmsx-1 and another ventralizing homeobox protein, Xvent-1, and show that Xmsx-1 is likely to be acting upstream of Xvent-1. We propose that Xmsx-1 is required in the BMP-stimulated ventralization pathway that involves the downstream activation of Xvent-1.
Assuntos
Padronização Corporal/fisiologia , Proteínas Morfogenéticas Ósseas/metabolismo , Proteínas de Homeodomínio/metabolismo , Proteínas Repressoras/metabolismo , Fatores de Transcrição , Proteínas de Xenopus , Animais , Ectoderma , Proteína Vmw65 do Vírus do Herpes Simples/genética , Proteínas de Homeodomínio/genética , Fator de Transcrição MSX1 , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Repressoras/genética , Transcrição Gênica , Xenopus/embriologiaRESUMO
Follistatin is expressed in Spemann's organizer in the Xenopus gastrula and mimics the activity of the organizer, inducing a neural fate directly in the ectoderm. We have previously shown that follistatin inhibits BMP activity through a direct interaction. In this study, we have characterized the localization and function of two follistatin isoforms to examine the functional differences between them. One notable difference, previously described, is that the shorter form (xFSS or xFS319) but not the C-terminally extended long form (xFSL) associates with cell-surface matrices. Here, we show that the spatial-temporal expression pattern of xFSL and xFSS is indistinguishable. Interestingly, however, xFSS was found to have a more potent inhibitory activity against BMP-4 than xFSL. Furthermore, using a surface plasmon resonance biosensor, xFSS was shown to have a higher binding capacity for BMP subtypes. The diffusion rates of xFSS and xFSL ectopically expressed in Xenopus embryos were similar. Taken together, our results suggest that the difference in BMP-inhibiting activity of the two follistatin isoforms is mainly attributable to a difference in their BMP binding properties rather than to their diffusion rates.
Assuntos
Glicoproteínas/metabolismo , Xenopus/embriologia , Xenopus/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas Morfogenéticas Ósseas/antagonistas & inibidores , Clonagem Molecular , Primers do DNA/genética , DNA Complementar/genética , Folistatina , Regulação da Expressão Gênica no Desenvolvimento , Glicoproteínas/genética , Dados de Sequência Molecular , Ligação Proteica , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Xenopus/genéticaRESUMO
PURPOSE: To determine the role of endothelin (ET)-ETA receptor mediation and endogenous production of endothelin-1 (ET-1) in the retinal response to hyperoxia. METHODS: Brown-Norway rats (n = 30) were injected intravitreally with an ETA receptor-selective antagonist, BQ-123, and an inhibitor of ET-converting enzyme (ECE), phosphoramidon, and were exposed to a 100% oxygen breathing mixture. Control rats underwent intravitreal injection of vehicle alone (2.5% Emulphor in phosphate-buffered saline). The retinal hemodynamic responses were analyzed using video-based fluorescein angiography (VFA) methodology. Baseline measurements were made with the animals breathing room air, and this was followed by intravitreal injections of the above agents. Subsequent VFA measurements were made after 5, 10, and 15 minutes of breathing 100% oxygen. RESULTS: The 10 rats injected with vehicle alone demonstrated the expected retinal response to hyperoxia, with significant (P < 0.001) vessel constriction (18% +/- 5%), an increase in retinal mean circulation time (0.84 +/- 0.13 seconds in room air and 1.59 +/- 0.27 seconds in 100% oxygen), and a decrease in blood flow (110.7 +/- 19.2 pixel2/second in room air and 41.9 +/- 9.0 pixel2/second in 100% oxygen), compared to values measured during room air breathing. The hyperoxic response in rats (n = 9) injected with 10(-4) M BQ-123 was significantly (P < 0.001) blunted compared to the group injected with vehicle alone. In contrast, intravitreal injection of saralasin, a specific angiotensin II receptor antagonist, had no significant effect on the retinal response to hyperoxia (n = 5). Intravitreal phosphoramidon (10(-3) M, n = 6) injection also resulted in a significantly (P < 0.001) blunted circulatory response to hyperoxia compared to rats injected with vehicle alone. This blunted response after ECE inhibition was comparable to that measured after ETA receptor antagonism with BQ-123 injection. CONCLUSIONS: These results demonstrate that the enhancement of ET-1 action, possibly caused by the activation of ECE, plays a primary role in regulating the retinal hemodynamic response to hyperoxia.
Assuntos
Circulação Sanguínea/fisiologia , Endotelinas/fisiologia , Hiperóxia/fisiopatologia , Receptores de Endotelina/fisiologia , Vasos Retinianos/fisiologia , Antagonistas de Receptores de Angiotensina , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Ácido Aspártico Endopeptidases/fisiologia , Velocidade do Fluxo Sanguíneo , Células Cultivadas , Antagonistas dos Receptores de Endotelina , Enzimas Conversoras de Endotelina , Endotelinas/genética , Endotélio Vascular/metabolismo , Angiofluoresceinografia , Expressão Gênica , Glicopeptídeos/farmacologia , Masculino , Metaloendopeptidases/antagonistas & inibidores , Metaloendopeptidases/fisiologia , Peptídeos Cíclicos/farmacologia , Inibidores de Proteases/farmacologia , Ratos , Ratos Endogâmicos BN , Receptores de Angiotensina/fisiologia , Respiração , Saralasina/farmacologia , Vasoconstrição/fisiologiaRESUMO
PURPOSE: To investigate the role of endogenous endothelin-1 (ET-1) expression and its interaction with the ETA receptor in the physiologic regulation of vascular tone as well as in the development of abnormal retinal hemodynamics in diabetes. METHODS: Retinal blood flow, using digitized video fluorescein angiography recordings, was quantitated after intravitreous injections of ET-1; BQ-123, an ETA receptor antagonist; and phosporamindon, an endothelin converting enzyme inhibitor in the eyes of diabetic and nondiabetic rats. A total of 154 rats were used for these experiments. Message levels of preproendothelin-1 (preproET-1) were measured from the retina of diabetic and nondiabetic rats using competitive polymerase chain reaction (PCR) techniques. RESULTS: Retinal blood flow was reduced (33%, P < 0.001) in diabetic rats compared to nondiabetic rats. BQ-123, an ETA receptor antagonist, but not saralasin, an angiotensin receptor antagonist, increased retinal blood flow in a dose-dependent manner in diabetic (EC50 of 8 x 10(-7) M) and in nondiabetic rats (EC50 of 8 x 10(-8) M). Besides being resistant to BQ-123, the maximal response in diabetic animals occurred 20 minutes later than in nondiabetic animals. Decreasing ET-1 levels by inhibiting endothelin-converting enzyme with phosphoramidon normalized retinal blood flow in diabetic rats. In nondiabetic rats, the intravitreous injection of exogenous ET-1 (10(-8) M) resulted in retinal blood flow decreases comparable to those measured in diabetic animals, and the subsequent injection of 10(-4) M BQ-123 produced retinal blood flow changes comparable to those measured in BQ-123 injected diabetic rats. Comparison of preproET-1 messenger RNA expression in the retina, brain and lung of control and diabetic rats using quantitative PCR and Northern blot analysis showed 2.0- and 1.7-fold increases in the retina and the brain, respectively, without changes in the lung. CONCLUSIONS: These data suggest that ET-1 is involved in the regulation of retinal blood flow in normal physiologic outcome, and an increase in the endogenous expression of ET-1 contributes to the reduction of retinal blood flow reported in the early stages of diabetes mellitus.
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Retinopatia Diabética/fisiopatologia , Endotelina-1/fisiologia , Vasos Retinianos/fisiologia , Animais , Encéfalo/metabolismo , Primers do DNA , Relação Dose-Resposta a Droga , Antagonistas dos Receptores de Endotelina , Endotelina-1/farmacologia , Endotelinas/genética , Endotelinas/metabolismo , Angiofluoresceinografia , Glicopeptídeos/farmacologia , Hemodinâmica/efeitos dos fármacos , Masculino , Metaloendopeptidases/antagonistas & inibidores , Peptídeos Cíclicos/farmacologia , Reação em Cadeia da Polimerase , Inibidores de Proteases/farmacologia , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos BN , Ratos Sprague-Dawley , Receptores de Endotelina/metabolismo , Retina/metabolismo , Vasos Retinianos/efeitos dos fármacosRESUMO
PURPOSE: Elevation of diacylglycerol (DAG) and protein kinase C (PKC) levels in diabetic vascular tissue is associated with abnormalities of retinal and renal hemodynamics. The object of this study was to determine whether direct elevation of retinal DAG levels, in the absence of diabetes or hyperglycemia, can mimic the hemodynamic abnormalities normally observed in diabetic rats. Retinal DAG levels were elevated using an inhibitor of DAG kinase that converts DAG to phosphatidic acid. The effectiveness of a specific PKC-beta isoform inhibitor introduced directly into the retinas of diabetic rats in reversing diabetes-related abnormal retinal hemodynamics was also investigated. METHODS: For retinal blood flow studies, diacylglycerol kinase (DGK) inhibitor R59949, at various concentrations, was injected into the vitreous of nondiabetic Sprague-Dawley rats (n = 33), and a PKC-beta isoform-selective inhibitor LY333531 was injected into the vitreous of rats with streptozotocin (STZ)-induced diabetes of 2 weeks' duration (n = 21). Retinal hemodynamic changes were quantitated using video-based fluorescein angiography. Total DAG levels were assayed from five nondiabetic rat retinas after DGK inhibition and retinal PKC activities were assayed from six diabetic rat retinas after PKC-beta inhibition. RESULTS: DGK inhibitor R59949 injected into the vitreous dose dependently increased the mean circulation time (MCT) and decreased retinal blood flow (EC50 = 10(-8) M). After 30 minutes, 10(-5) M R59949 induced a 1.7-fold increase in total retinal DAG levels, compared with the levels in vehicle-injected eyes, an increase in MCT from 0.87 +/- 0.05 seconds to 1.44 +/- 0.12 seconds (P < 0.01) and a decrease in retinal blood flow from 105.3 +/- 6.5 pixel2/second to 64.1 +/- 5 pixel2/second (P < 0.01). The effect of R59949 was sustained for 60 minutes after injection. These retinal hemodynamic parameters after DGK inhibition were comparable to those measured at baseline in rats with STZ-induced diabetes of 2 weeks' duration (MCT = 1.38 +/- 0.20 seconds; retinal blood flow = 68 +/- 11.2 pixel2/second). Intravitreal injection of the PKC-beta inhibitor (LY333531) at 10(-5) M in diabetic rats decreased by a factor of 1.6 the diabetes-related increased PKC activation, decreased the prolonged MCT (0.98 +/- 0.13 seconds; P < 0.01) and increased retinal blood flow (93.4 +/- 14.2 pixel2/second; P < 0.01). The measured retinal circulatory parameters after PKC inhibition in the retina were comparable to those measured at baseline in the nondiabetic rats. CONCLUSIONS: These results provide direct evidence that DAG elevation and subsequent PKC-beta isoform activation are the primary biochemical sequelae responsible for the development of the abnormal retinal hemodynamics observed in diabetic rats.
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Diglicerídeos/metabolismo , Isoenzimas/metabolismo , Proteína Quinase C/metabolismo , Retina/metabolismo , Vasos Retinianos/fisiopatologia , Animais , Velocidade do Fluxo Sanguíneo/fisiologia , Diabetes Mellitus Experimental/metabolismo , Diacilglicerol Quinase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Angiofluoresceinografia , Hiperglicemia/metabolismo , Hiperglicemia/fisiopatologia , Indóis/farmacologia , Isoenzimas/antagonistas & inibidores , Masculino , Maleimidas/farmacologia , Piperidinas/farmacologia , Proteína Quinase C/antagonistas & inibidores , Quinazolinas/farmacologia , Quinazolinonas , Ratos , Ratos Sprague-Dawley , Retina/efeitos dos fármacos , Retina/fisiopatologiaRESUMO
PURPOSE: To investigate the distribution of inflammatory mediators such as interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha and angiogenic cytokines such as vascular endothelial growth factor (VEGF) and to identify their cellular source in surgically excised choroidal neovascular membranes (CNVMs) of various origins. METHODS: Immunoperoxidase staining was performed on paraffin-embedded sections of 11 surgically excised CNVMs to identify cellular distribution and localization of cytokines. Immunofluorescent double staining was performed to detect the cellular source of cytokines. RESULTS: Cytokeratin-positive cells were detected in the RPE layer, in stromal cells, and around neovascular vessels. Macrophages identified by their cellular marker CD68 showed almost the same distribution as cytokeratin-positive cells, although they were most prominent in the stroma. A substantial number of neovascular vessels were also immunoreactive to IL-1beta and TNF-alpha. Immunofluorescent double staining revealed that the RPE layers immunopositive for cytokeratin were also immunopositive for all cytokines, whereas stromal cells immunostained for CD68 were positive for IL-1beta and TNF-alpha, but not for VEGF. CONCLUSIONS: These results indicate that IL-1beta and TNF-alpha secreted by macrophages may promote, at least in part, angiogenesis in CNVMs by stimulating VEGF production in RPE cells.
Assuntos
Neovascularização de Coroide/etiologia , Neovascularização de Coroide/metabolismo , Fatores de Crescimento Endotelial/metabolismo , Interleucina-1/metabolismo , Linfocinas/metabolismo , Macrófagos/fisiologia , Fator de Necrose Tumoral alfa/metabolismo , Adulto , Idoso , Neovascularização de Coroide/patologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Técnicas Imunoenzimáticas , Masculino , Epitélio Pigmentado Ocular/metabolismo , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularRESUMO
The constants of the overall extraction equilibrium (K(ex)), the partition for various diluents having low dielectric constants (K(D,MLA)), the aqueous ion-pair formation (K(MLA)), and the dimer formation in CCl(4) of 16-crown-5 (16C5)-alkali metal (Na, K) picrate 1:1:1 complexes were determined at 25 degrees C; the distribution constants of 16C5 were also measured at 25 degrees C. The logK(MLA) of Na and K are 4.14+/-0.19 and 3.05+/-0.28, respectively. The partition behavior of 16C5 and its 1:1:1 complexes with the alkalimetal picrates can be explained by regular solution theory, except for CHCl(3); the molar volumes and solubility parameters of 16C5 and the 1:1:1 complexes were determined. The magnitude of K(ex) largely depends on that of K(MLA). For every diluent, 16C5 always shows Na(+) extraction-selectivity over K(+). The K(MLA) value most contributes to the extraction selectivity of 16C5 for Na(+) over for K(+) among the three fundamental equilibrium constants, the aqueous 1:1 complex-formation constant of 16C5 with the alkali metal ion, K(MLA), and K(D,MLA). Furthermore, correct contributions of a methylene group to distribution constants of organic compounds between diluents of low dielectric constants and water were determined by the distribution constants of 16C5 and 15-crown-5; the additivity of the contributions of functional groups to the partition constant of a crown ether was verified.
RESUMO
Acarbose is an inhibitor of intestinal alpha-glucosidase and has been reported to decrease blood glucose concentrations and glycosuria in diabetic patients and animals. In this study we investigated whether this drug could prevent the abnormalities detected in retinal circulation of diabetic rats. Longitudinal paired studies were performed and the changes in retinal circulation were analyzed using video based fluorescein angiography (VFA) methodology in the same animal. Baseline VFA recordings were obtained from 41 rats. These rats were separated into 4 different groups: In group A (n = 12), diabetes was induced by streptozotocin (STZ) injection and the rats were fed with acarbose (40 mg/100 g powdered chow) mixed into regular rat chow; In group B (n = 10), diabetes was induced by STZ injection and the rats were fed with normal chow; In group C (n = 9), the non-diabetic rats were fed with acarbose; In group D (n = 10), the non-diabetic rats were fed with normal chow. At the end of 2 weeks, all rats again underwent VFA recordings. Blood glucose levels and body weights of rats were monitored during the experiment. The mean blood glucose concentration of Group B was raised from 98.5 +/- 8.7 to 342 +/- 30 mg/dl after STZ injection while in Group A, this change in glucose level was partially ameliorated by acarbose (from 102 +/- 15 to 247 +/- 48 mg/dl). In Group C and D, the blood glucose levels were not significantly changed during the experiment.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Diabetes Mellitus Experimental/fisiopatologia , Inibidores Enzimáticos/farmacologia , Inibidores de Glicosídeo Hidrolases , Vasos Retinianos/fisiopatologia , Trissacarídeos/farmacologia , Acarbose , Animais , Glicemia/análise , Diabetes Mellitus Experimental/tratamento farmacológico , Hemodinâmica/efeitos dos fármacos , Estudos Longitudinais , Masculino , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional/efeitos dos fármacosRESUMO
Mean (+/- SD) serum erythropoietin (EPO) levels were 18.6 +/- 5.6 mU/ml in 180 normal Japanese subjects. Serum EPO levels were elevated with a negative correlation on a log scale (r = -0.864, P < 0.005) to hematocrit (Ht) values in anemic patients not associated with rheumatoid arthritis (RA) or chronic renal failure (CRF). Serum EPO levels in patients with RA (31.6 +/- 16.4 mU/ml) were relatively lower than those in normal subjects and anemic patients without RA or CRF when matched for comparative Ht values. Seven anemic patients with RA were treated by daily subcutaneous (sc) injection of recombinant EPO (rEPO, 500-1,000 U/day) for 4 weeks. The patients had initial Ht values of 25.1% or less and maintained stable clinical status. The treatment with rEPO raised serum EPO levels (53.8 +/- 15.2 mU/ml, P < 0.05), which resulted in an increase in Ht values (more than 3%) in 6 out of 7 patients with RA. The mean (+/- SD) Ht values at the end of the treatment with rEPO (500-1,000 U/day) were greater than those before the treatment in the 7 patients with RA (28.5 +/- 4.6 vs. 22.7 +/- 2.5%, P < 0.05). These findings suggest that chronic anemia associated with RA may be corrected by daily sc injection of a small dose of rEPO.
Assuntos
Anemia/tratamento farmacológico , Artrite Reumatoide/complicações , Eritropoetina/administração & dosagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Anemia/complicações , Criança , Pré-Escolar , Doença Crônica , Eritropoetina/sangue , Eritropoetina/uso terapêutico , Feminino , Humanos , Injeções Subcutâneas , Masculino , Pessoa de Meia-Idade , Radioimunoensaio , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/sangue , Proteínas Recombinantes/uso terapêutico , Fatores de TempoRESUMO
In February, 1990, a 49-year-old man was admitted with petechia and gingival bleeding. The peripheral blood showed 5,200 leukocytes/microliters including 73% abnormal promyelocytes and 24,000/microliters platelets. Bone marrow puncture revealed that nucleated cell count was 331,250/microliters including 85.4% abnormal promyelocytes with 46XY, i(17q) chromosome. Coagulation tests revealed DIC. He was diagnosed as having acute promyelocytic leukemia, and he was treated with the BHAC-DMP protocol. He achieved complete remission, and received consolidation therapy and maintenance therapy. However, he relapsed in May, 1991 with 46XY, 16q-, i (17q) chromosome. He was treated with BHAC-MV protocol and again achieved complete remission. In June, 1992, he re-relapsed and 3.6% blasts and 10% abnormal promyelocytes was found in his bone marrow. He was treated for 14 days with 15 mg Aclarubicin without any change. Then he was treated with 60 mg All-trans retinoic acid (ATRA). After administration of ATRA, his peripheral blood leukocyte count increased temporarily but bone marrow suppression continued. Then he received continuous subcutaneous infusion of 24 micrograms/day granulocyte colony-stimulating factor (rhG-CSF). After treatment with ATRA and rhG-CSF, he entered a third complete remission.
Assuntos
Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Leucemia Promielocítica Aguda/terapia , Tretinoína/uso terapêutico , Terapia Combinada , Humanos , Leucemia Promielocítica Aguda/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Recidiva , Indução de RemissãoRESUMO
A 78-year-old female suffered from idiopathic paroxysmal cold hemoglobinuria (PCH). Her symptom occurred immediately after she worked outside in the cold early morning. This case is characterized by the eldest case with non-syphilitic PCH reported in Japan and by the most advanced anemia (Hb 4.2 g/dl) possibly due to prolonged hemolysis after cold exposure. Analysis of the serum revealed positive Donath-Landsteiner antibody of IgG type, which could react to hemolysis not only below 15 degrees C but also at 15-20 degrees C in vitro.
Assuntos
Temperatura Baixa/efeitos adversos , Hemoglobinúria Paroxística/etiologia , Idoso , Autoanticorpos/análise , Feminino , Hemoglobinúria Paroxística/imunologia , Hemólise , Humanos , Imunoglobulina G/análiseRESUMO
Subcutaneous administration of recombinant human Interleukin-1 beta (IL-1 beta) in a dose of 1-3 x 10(4) U/day for 14 to 72 days resulted in an increase in circulating granulocytes and bone marrow monocytes in all the 4 patients examined. Circulating platelet count was also increased in two of four patients with myelodysplastic syndrome (MDS) and aplastic anemia (AA). Bone marrow examination revealed an increase in megakaryocyte count in these patients, whereas the percentage of blast was not changed. An increase in blood platelet count was accompanied by an increase in serum GM-CSF in a patient with AA, whereas serum IL-6 level was not changed throughout the treatment with IL-1 beta. These findings suggest that IL-1 beta may be useful for the treatment of a proportion of patients with MDS and AA who are associated with thrombocytopenia.
Assuntos
Anemia Aplástica/terapia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/sangue , Interleucina-1/farmacologia , Síndromes Mielodisplásicas/terapia , Contagem de Plaquetas/efeitos dos fármacos , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Injeções Subcutâneas , Interleucina-1/administração & dosagem , Masculino , Pessoa de Meia-IdadeRESUMO
Serum erythropoietin (EPO) levels were determined by the recombigen EPO RIA kit (DPC) in normal subjects and patients with renal dysfunction, diabetes mellitus, hypothyroidism and a variety of hematological disorders. Mean (+/- SD) serum EPO levels were 18.6 +/- 5.6 mU/ml in 180 normal subjects and no sex difference was obtained. Serum EPO levels in older subjects were slightly greater than those in younger subjects. There was a negative correlation between serum EPO levels and Ht values in anemic patients with normal renal function, whereas serum EPO levels were within the normal range in anemic patients with renal disorders, suggesting that serum EPO levels were relatively low in patients with chronic renal failure. Serum EPO levels were rather increased in patients with diabetes mellitus and hypothyroidism. High serum EPO levels were obtained in patients with a variety of hematological disorders such as acute leukemia, multiple myeloma, myelodysplasia syndrome, aplastic anemia and pure red cell aplasia. In a patient with pure red cell aplasia treated with glucocorticoids, serum EPO levels were lowered before anemia was recovered and reticulocytes were increased. These findings indicate that measurement of serum EPO levels are useful for not only differential diagnosis of anemia but also clinical evaluation of the treatment.
Assuntos
Anemia/sangue , Diabetes Mellitus/sangue , Eritropoetina/sangue , Kit de Reagentes para Diagnóstico , Aplasia Pura de Série Vermelha/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valores de ReferênciaRESUMO
A 57-year-old female presented with general fatigue. She had neither lymphadenopathy nor hepatosplenomegaly. Laboratory data revealed anemia and leukopenia (1,500/microliters) with a differential count of 4.5% leukemic cells. The myelogram revealed 34.4% leukemic cells, of which diameter ranged from 20 to 28 microns. The diagnosis was acute myelogenous leukemia (FAB: M2) with myelodysplasia. Cytogenetic analysis revealed that the leukemic cells had chromosome abnormalities involving both diploidy and tetraploidy with structural rearrangement. Structural rearrangement included del(5) (q22q33), del(15) (q22q24), and t(3; 12) (q25;p13). Small dose aclacinomycin-A treatment was effective in reducing the number of leukemic cells in bone marrow, and both anemia and leukocytopenia were improved.
Assuntos
Aberrações Cromossômicas , Leucemia Mieloide Aguda/genética , Síndromes Mielodisplásicas/patologia , Poliploidia , Aclarubicina/administração & dosagem , Feminino , Humanos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/patologia , Pessoa de Meia-IdadeRESUMO
Hemolytic uremic syndrome (HUS) is caused by endothelial cell damages. Ninety percent of children with HUS have verotoxin-producing E.coli infection. Verotoxin binds to glycolipid receptors globotriaosyl ceramide (Gb3), and the difference of Gb3 expression level in each organ would lead to specific organ involvement. The receptors are expressed in human renal cortex and medulla. The expression level of Gb3 in normal human brain has not been characterized completely. However involvement of central nervous system is a severe complication of HUS. Spreading of microvascular thrombosis caused by combined effects of lipopolysaccharide, cytokine, enhanced shear stress, and verotoxin would play a major role in the development of central nervous dysfunction.
Assuntos
Toxinas Bacterianas/toxicidade , Doenças do Sistema Nervoso Central/etiologia , Infecções por Escherichia coli , Escherichia coli O157 , Síndrome Hemolítico-Urêmica/etiologia , Toxinas Bacterianas/metabolismo , Hemorragia Gastrointestinal/etiologia , Humanos , Púrpura Trombocitopênica Trombótica/etiologia , Toxina Shiga I , Triexosilceramidas/metabolismoRESUMO
INTRODUCTION: Although it has been established that hyperthyroidism leads to reduced bone mineral density (BMD), with accelerated bone turnover promoting bone resorption in female patients, there is a dearth of data for male patients with hyperthyroidism. This study evaluated BMD and bone metabolism in male patients with Graves' disease. METHODS: The study included 56 Japanese male patients with newly diagnosed Graves' disease and 34 normal Japanese male control subjects of similar age and body mass index. We used dual energy x-ray absorptiometry to measure BMD at sites with different cortical/cancellous bone ratios (lumbar spine, femoral neck, and distal radius). RESULTS: At the lumbar spine and the distal radius, BMD and T-scores were significantly lower for patients than for controls. At the femoral neck, on the other hand, the same values were relatively, but not significantly, lower in patients than in controls. However, Z-scores at all three sites were significantly lower for patients than for controls. The Z -score at the distal radius of patients was significantly lower than that at their lumbar spine and femoral neck. In addition, Z-score at the distal radius correlated negatively with age, free thyroxine, thyroid stimulating hormone receptor antibodies, thyroid stimulating antibody, and urinary N-terminal telopeptide of type I collagen normalized by creatinine. CONCLUSIONS: These results indicate a high prevalence of cortical bone loss in male patients with Graves' disease, especially elderly patients. We conclude that BMD measurement is crucial in all Graves' disease patients regardless of their gender and that the radial BMD as well as BMD at the lumbar spine and femoral neck should be monitored to effectively prevent bone loss and subsequent fracture.