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1.
Science ; 162(3849): 127-9, 1968 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-4970764

RESUMO

Antiserums to human prothrontbin contain antibodies directed against at least two different antigenic sites. During blood coagulation, prothrombin is cleaved into two major antigenically distinct fragments-thrombin and a "pro" fragment. The latter is present in normal seruim, whereas thrombin loses its immunologic reactivity, presumably because of its combination with the natural thrombin inhibitors in blood.


Assuntos
Coagulação Sanguínea , Soros Imunes , Protrombina , Reações Antígeno-Anticorpo , Humanos , Imunoquímica , Imunodifusão , Imunoeletroforese
2.
Science ; 160(3829): 786-7, 1968 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-4171540

RESUMO

Seven new cases of acquired inhibitors of Factor VIII have been typed immunologically as (gamma2kappa2) antibodies. Electrophoretic mobility on starch block of a combined group of 13 such antibodies is considerably more rapid than that of the bulk of immunoglobulin G.


Assuntos
Anticorpos/análise , Fator VIII/antagonistas & inibidores , Eletroforese das Proteínas Sanguíneas , Hemofilia A/imunologia , Transtornos Hemorrágicos/imunologia , Humanos , Peptídeos/análise , Amido , gama-Globulinas/análise
3.
Science ; 282(5391): 1145-7, 1998 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-9804556

RESUMO

Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages. After undifferentiated proliferation in vitro for 4 to 5 months, these cells still maintained the developmental potential to form trophoblast and derivatives of all three embryonic germ layers, including gut epithelium (endoderm); cartilage, bone, smooth muscle, and striated muscle (mesoderm); and neural epithelium, embryonic ganglia, and stratified squamous epithelium (ectoderm). These cell lines should be useful in human developmental biology, drug discovery, and transplantation medicine.


Assuntos
Blastocisto/citologia , Técnicas de Cultura de Células , Linhagem Celular , Células-Tronco/citologia , Animais , Antígenos Glicosídicos Associados a Tumores , Diferenciação Celular , Criopreservação , Ectoderma/citologia , Endoderma/citologia , Feminino , Glicoesfingolipídeos/análise , Rejeição de Enxerto , Humanos , Cariotipagem , Masculino , Mesoderma/citologia , Camundongos , Camundongos SCID , Antígenos Embrionários Estágio-Específicos , Transplante de Células-Tronco , Células-Tronco/química , Telomerase/metabolismo , Teratoma/etiologia , Trofoblastos/citologia
4.
J Clin Invest ; 46(2): 147-56, 1967 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-6018754

RESUMO

Investigations were undertaken of the chemical nature and kinetics of interaction of three acquired inhibitors of Factor VIII. The inhibition was stoichiometric, one molecule of inhibitor (or one site on a molecule) being required to inactivate one molecule of Factor VIII. All three inhibitors were found to be monotypic antibodies of class IgG, Type K (gamma(2) kappa(2)). This appears to be the second syndrome due to the production of monotypic antibody, and the first instance involving IgG immunoglobulins.


Assuntos
Fator VIII/análise , Fator VIII/metabolismo , Renina/sangue , Adolescente , Adulto , Formação de Anticorpos , Colite Ulcerativa/imunologia , Feminino , Hemofilia A/imunologia , Transtornos Hemorrágicos/imunologia , Humanos , Imunoeletroforese , Cinética , Masculino , Temperatura
5.
J Clin Invest ; 68(2): 321-8, 1981 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-6790574

RESUMO

Human Factor VIII desialylated by treatment with Vibrio cholerae neuraminidase (ASVIII) aggregated human platelets in the absence of ristocetin in platelet-rich plasma and, to a lesser extent, in washed platelet suspensions. Aggregation is accompanied by thromboxane formation and is completely inhibited by EDTA. Aspirin blocks the second phase of aggregation and abolishes thromboxane production. Subaggregating doses of ASVIII and of either ADP, epinephrine, or collagen produce prompt and complete platelet aggregation. Bernard-Soulier syndrome platelets either did not aggregate with ASVIII (Two cases) or showed markedly decreased aggregation (one cases). Factor VIII complex was prepared from the plasma of two patients with variant von Willebrand's disease (sialic acid content 142 and 75 nmol/mg, respectively); neither protein generated platelet-aggregating activity upon desialylation. [3H]ASVIII binds rapidly to platelets and 37 degrees C, while tritiated, fully sialylated factor VIII binds to a negligible extent. As little as 1--2 micrograms ASVIII bound/10(9) platelets is capable of inducing platelet aggregation. ASVIII may be a useful tool for investigating platelet-Factor VIII interactions in the absence of ristocetin. Furthermore, desialylated Factor VIII might play a physiologic role in Factor VIII-mediated platelet reactions in vivo.


Assuntos
Assialoglicoproteínas , Fator VIII/análogos & derivados , Agregação Plaquetária/efeitos dos fármacos , Púrpura Trombocitopênica/sangue , Plaquetas/metabolismo , Fator VIII/farmacologia , Humanos , Ligação Proteica , Ristocetina/farmacologia , Ácidos Siálicos/sangue , Relação Estrutura-Atividade , Síndrome
6.
J Clin Invest ; 48(7): 1292-8, 1969 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-5794251

RESUMO

The metabolism of human prothrombin labeled with radioactive iodine was studied in seven normal subjects and four hemophilic patients. Results in the normal subjects were: plasma volume, 37.6+/-3.8 ml/kg; plasma prothrombin concentration, 303+/-40 U/ml (0.153+/-0.02 mg/ml); prothrombin half-life, 2.81+/-0.51 days; total plasma prothrombin pool, 5.72+/-0.62 mg/kg, representing 64.1+/-9.1% of total body prothrombin; fractional catabolic rate, 42.5+/-12.4% of the plasma pool per day; prothrombin synthesis rate, 2.43+/-0.76 mg/kg per day. Results in the hemophilic patients did not differ significantly from normal. Circulating products of prothrombin activation could not be demonstrated in normal individuals or hemophilic subjects. The data suggest that continuous physiologic activation of the blood coagulation mechanism plays only a small part, if any, in the normal catabolism of prothrombin.


Assuntos
Hemofilia A/metabolismo , Hemofilia B/metabolismo , Protrombina/metabolismo , Adulto , Coagulação Sanguínea , Cromatografia em Gel , Eletroforese , Humanos , Injeções Intravenosas , Isótopos de Iodo , Masculino , Pessoa de Meia-Idade , Volume Plasmático
7.
J Clin Invest ; 48(12): 2251-9, 1969 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-5355338

RESUMO

A large family has been studied, 11 of whose members have half-normal plasma concentrations of biological prothrombin activity. The pattern of inheritance is autosomal. By use of a specific immunoassay, affected family members have been shown to possess normal quantities of immunoreactive prothrombin, whose immunologic properties seem identical with those of the normal zymogen. Prothrombin isolation from the plasma of one such individual gave normal yields of protein but half-normal amounts of prothrombin activity. Activation of this material in the "intrinsic" and "extrinsic" systems, in concentrated sodium citrate, or by trypsin, gives rise to half, or less, of the thrombin clotting and esterase activities expected from a comparable normal prothrombin preparation. During the clotting of blood from an affected individual, all material with the mobility of prothrombin disappears. Immunoelectrophoresis of the serum reveals a normal nonthrombin "pro piece," and an additional activation product with an electrophoretic mobility intermediate between that of prothrombin and of "pro piece." These results suggest that affected individuals are heterozygotes in whom half the prothrombin molecules synthesized are structurally abnormal, since they undergo some alterations during activation, but are incapable of releasing the active enzyme, thrombin.


Assuntos
Hipoprotrombinemias/genética , Adolescente , Adulto , Criança , Aberrações Cromossômicas , Transtornos Cromossômicos , Feminino , Humanos , Imunoeletroforese , Isótopos de Iodo , Masculino , Linhagem , Protrombina/sangue , Tempo de Protrombina
8.
J Clin Invest ; 66(3): 397-405, 1980 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-6772673

RESUMO

Prolongation of all phospholipid-dependent coagulation tests was found in a patient with macroglobulinemia, despite absence of bleeding manifestations. The purified monoclonal IgM lambda protein and its Fabmu tryptic fragment induced similar changes in normal plasma. Patient IgM and Fabmu completely inhibited Ca++-dependent binding of radiolabeled prothrombin and Factor X to mixed phospholipid micelles. The patient's IgM lambda paraprotein reacted with phosphatidylserine and, to a lesser extent, with phosphatidylinositol and phosphatidic acid, but not with phosphatidylcholine or phosphatidylethanolamine. Prior incubation of phospholipid with patient Fabmu blocked the positive reactions. Substitution of washed platelets for phospholipid led to normalization of patient coagulation tests and corrected all abnormalities produced in normal plasma by patient IgM. Furthermore, binding of 125I-Factor Xa to thrombin-treated platelets was entirely normal in the presence of patient IgM. These studies support the concept that platelets, rather than phospholipid micelles, are the primary locus of prothrombin and Factor X activation in normal hemostasis.


Assuntos
Coagulação Sanguínea , Cadeias Leves de Imunoglobulina/fisiologia , Imunoglobulina M/fisiologia , Cadeias lambda de Imunoglobulina/fisiologia , Lúpus Eritematoso Sistêmico/sangue , Fosfolipídeos/imunologia , Especificidade de Anticorpos , Plaquetas/fisiologia , Fator X/metabolismo , Humanos , Lúpus Eritematoso Sistêmico/imunologia , Fosfolipídeos/sangue , Ligação Proteica , Protrombina/metabolismo , Macroglobulinemia de Waldenstrom/sangue , Macroglobulinemia de Waldenstrom/imunologia
9.
J Clin Invest ; 75(3): 896-901, 1985 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-3156882

RESUMO

We have previously described a series of monoclonal antibodies against platelet membrane glycoproteins. Two of the antibodies, B59.2 and B2.12, recognize the glycoprotein IIb-IIIa complex. These two antibodies react specifically with glycoprotein (GP) IIIa, as shown by immunoblotting of sodium dodecyl sulfate-polyacrylamide gels of solubilized platelet membranes. Monoclonal B2.12, but not B59.2, binds to cultured human endothelial cells obtained from umbilical vein, internal iliac artery, and inferior vena cava. At saturation approximately 100,000 binding sites were detected per human umbilical vein endothelial cell. When solubilized radioiodinated cells were chromatographed on a column of agarose-bound B2.12, a single radiolabeled protein was obtained whose apparent molecular weight is slightly larger than that of platelet GP IIIa. This protein incorporated [35S]methionine when endothelial cells were labeled metabolically. These results demonstrate that human endothelial cell membranes synthesize a protein immunologically related to platelet GP IIIa.


Assuntos
Anticorpos Monoclonais , Plaquetas/imunologia , Endotélio/citologia , Glicoproteínas/imunologia , Proteínas de Membrana/imunologia , Aorta , Sítios de Ligação de Anticorpos , Plaquetas/metabolismo , Células Cultivadas , Cromatografia de Afinidade , Endotélio/imunologia , Endotélio/metabolismo , Glicoproteínas/biossíntese , Glicoproteínas/isolamento & purificação , Humanos , Soros Imunes/farmacologia , Proteínas de Membrana/biossíntese , Proteínas de Membrana/isolamento & purificação , Peso Molecular , Glicoproteínas da Membrana de Plaquetas , Veias Umbilicais
10.
J Clin Invest ; 53(2): 600-11, 1974 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11344575

RESUMO

A new, autosomally inherited abnormal fibrinogen associated with hypofibrinogenemia has been described in several members of a family. Plasma fibrinogen measured either as thrombin-clottable protein or by immunodiffusion revealed a fibrinogen level ranging between 60 and 90 mg/100 ml. The thrombin time of plasma or purified fibrinogen was prolonged and only partially corrected by the addition of calcium. Purified fibrinogen prolonged the thrombin time of normal plasma. Fibrinopeptide release by thrombin was normal in rate and amount, but fibrin monomer aggregation was grossly disturbed, especially in a high ionic strength medium. We have designated this fibrinogen "fibrinogen Philadelphia." Acrylamide gel electrophoresis of mixtures of [121I]normal and [125I]abnormal fibrinogens revealed a slight increase in the anodal mobility of fibrinogen Philadelphia. Similarly, DEAE-cellulose chromatography showed slightly stronger binding of fibrinogen Philadelphia than normal. To elucidate the mechanism responsible for the low plasma fibrinogen concentration, simultaneous metabolic studies of autologous (patient) and homologous (normal) fibrinogen, labeled with 125I and 121I, respectively, were performed in two affected subjects. Autologous fibrinogen half-life was short and the fractional catabolic rate was markedly increased in both family members. In contrast, homologous fibrinogen half-life and fractional catabolic rate were normal. These metabolic studies demonstrate that rapid degradation of fibrinogen Philadelphia is largely responsible for the depressed levels of a plasma fibrinogen. This represents the first example of a mutant plasma protein in which the molecular defect is associated with an altered catabolism.


Assuntos
Afibrinogenemia/metabolismo , Fibrinogênios Anormais/metabolismo , Adolescente , Adulto , Afibrinogenemia/fisiopatologia , Coagulação Sanguínea , Cromatografia DEAE-Celulose/métodos , Eletroforese em Gel de Poliacrilamida/métodos , Feminino , Fibrina/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Linhagem , Tempo de Protrombina , Tempo de Trombina
11.
J Clin Invest ; 93(6): 2417-24, 1994 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8200976

RESUMO

Glycoprotein Ib beta (GPIb beta) exists in platelets disulfide-linked to glycoprotein Ib alpha (GPIb alpha), a major receptor for von Willebrand factor. Both GPIb alpha and GPIb beta are expressed in endothelial cells (EC). While the GPIb alpha mRNA and protein appear similar in platelets and EC, EC GPIb beta mRNA is larger than platelet GPIb beta and encodes a larger protein. We have cloned and sequenced EC GPIb beta cDNA and report a 2793-nucleotide sequence which contains a 411-amino acid open reading frame. The EC sequence contains all of the platelet cDNA sequence and all but three amino acids of the primary translation product. Like the genes encoding GPIb alpha, GPIX, and GPV, the GPIb beta gene appears simple in structure. Using human hamster hybrids, we have localized the GPIb beta gene to chromosome 22pter-->22q11.2. When we examined poly (A)+ RNA from several human tissues for GPIb beta mRNA expression, we found that GPIb beta mRNA was expressed in a variety of tissues but was most abundant in heart and brain, while GPIb alpha and GPIX mRNA expression was found only in lung and placenta at very low levels. The broad distribution of GPIb beta mRNA suggests that it may be playing a role different than or additional to its function in platelets.


Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 22 , DNA Complementar/isolamento & purificação , Endotélio Vascular/química , Glicoproteínas da Membrana de Plaquetas/genética , Sequência de Aminoácidos , Sequência de Bases , Células Cultivadas , Clonagem Molecular , DNA Complementar/química , Humanos , Dados de Sequência Molecular , Glicoproteínas da Membrana de Plaquetas/análise , RNA Mensageiro/análise
12.
Cancer Res ; 36(10): 3702-6, 1976 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-133754

RESUMO

Synthetic aromatic analogs of retinoic acid were administered i.p. and p.o. to Fischer F344 rats bearing a transplantable chondrosarcoma. 35CO4 incorporation into glycosaminoglycans were compared for neoplastic and normal cartilage explants after removal from animals given various analogs. There was a direct relationship between [35S]glycosaminoglycan synthesis by chondrosarcoma chondrocytes and inhibition of tumor growth. The degree of inhibition of [35S]glycosaminoglycan synthesis in the neoplastic cartilage was dependent on the dose of the retinoid administered. At 20-mg/kg/day doses of retinoid for 4 weeks, 35SO4 incorporated into glycosaminoglycan by treated tumor explants was reduced as much as 95%. There was no reduction of [35S] glycosaminoglycan produced in normal costal cartilage of the same animals. Retinoid treatment of 20-mg/kg/day doses for 4 weeks resulted in a 75% reduction in glycosaminoglycan per mg of chondrosarcoma; there was no reduction in costal cartilage glycosaminoglycan. Retinoid (10- to 20-mg/kg/day doses) elevated collagen levels per mg of chondrosarcoma but had no effect on costal cartilage collagen. Combined in vitro and in vivo studies showed that retinoid administration modified neoplastic chondrocyte function but had no measurable effect on normal chondrocyte function.


Assuntos
Condrossarcoma/metabolismo , Glicosaminoglicanos/biossíntese , Tretinoína/análogos & derivados , Vitamina A/análogos & derivados , Animais , Cartilagem/metabolismo , Colágeno/metabolismo , Relação Dose-Resposta a Droga , Ratos , Ratos Endogâmicos F344 , Sarcoma Experimental/metabolismo , Sulfatos/metabolismo , Tretinoína/farmacologia
13.
Biochim Biophys Acta ; 385(2): 221-31, 1975 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-123777

RESUMO

The sulfation of glycosaminoglycans by ascorbic acid 2-[35S]sulfate was studied in costal cartilage and chondrocytes in vitro. Negligable (if any) sulfation of glycosaminoglycans was detected with immediately isolated ascorbic acid 2-[35S]sulfate. However, formation of [35S]glycosaminoglycans was readily detected with ascorbic acid 2-[35S]sulfate which had been stored at minus 20 degrees C for several days. The [35S]glycosaminoglycans did not result from the direct transfer of 35S from ascorbic acid 2-sulfate but rather from a decomposition product of ascorbic acid 2-[35S]sulfate. Evidence is presented to show that the sulfation pathway with the decomposition product involves exchange with inorganic sulfate, and strongly suggests that sulfation proceeds via 3'-phosphoadenosine 5'-phosphosulfate. The decomposition product appears similar to inorganic sulfate in several test systems. In view of these observations, it is suggested that previous conclusions implicating as acid 2-sulfate as a biological sulfate donor, based on the use of ascorbic acid 2-[35S]sulfate be re-evaluated.


Assuntos
Ácido Ascórbico/metabolismo , Cartilagem/metabolismo , Glicosaminoglicanos/biossíntese , Sulfatos/metabolismo , Nucleotídeos de Adenina/metabolismo , Animais , Cartilagem/efeitos dos fármacos , Estabilidade de Medicamentos , Estudos de Avaliação como Assunto , Álcoois Graxos/metabolismo , Masculino , Ratos , Costelas , Sulfatos/farmacologia , Radioisótopos de Enxofre
14.
Biochim Biophys Acta ; 924(1): 127-34, 1987 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-3470050

RESUMO

The human erythroleukemia (HEL) cell line is known to express a number of platelet-megakaryocyte markers, including glycoproteins IIb and IIIa. Using [35S]methionine as well as monoclonal and polyclonal antibodies to glycoprotein IIb and IIIa, we have demonstrated synthesis of these two glycoproteins by this cell line. When Triton X-100-solubilized membranes were subjected to cross immunoelectrophoresis in 1% agarose, using a mixture of polyclonal antibodies to glycoproteins IIb and IIIa, a single peak was seen in the presence of Ca2+, whereas two distinct peaks were seen in the presence of 5 mM EDTA. When these crossed immunoelectrophoresis were overlaid with 125I-labelled fibrinogen, binding of fibrinogen to the glycoprotein IIb and IIIa complex peak was observed. When either the electrophoresis or the subsequent overlay was done in the presence of 5 mM EDTA, no 125I-fibrinogen binding was observed. These studies demonstrate that HEL cells contain glycoproteins IIb and IIIa capable of forming a Ca2+-dependent complex and capable of binding fibrinogen in a Ca2+-dependent reaction. Nevertheless, glycoprotein IIb- and IIIa-specific fibrinogen binding, of the type observed in platelets, could not be demonstrated in 'resting' or 'stimulated' intact HEL cells. The mechanism giving rise to this difference is currently unknown.


Assuntos
Fibrinogênio/metabolismo , Glicoproteínas da Membrana de Plaquetas/biossíntese , Anticorpos , Anticorpos Monoclonais , Plaquetas/metabolismo , Linhagem Celular , Humanos , Imunoeletroforese Bidimensional , Cinética , Leucemia Eritroblástica Aguda , Glicoproteínas da Membrana de Plaquetas/metabolismo , Ligação Proteica
15.
Biochim Biophys Acta ; 429(2): 508-16, 1976 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-4134

RESUMO

Pure human arylsulfatase A (EC 3.1.6.1) was found to hydrolyze ascorbic acid 2-sulfate to ascorbic acid and inorganic sulfate at rates from 200 to 2000 mumol/mg per h depending on the method of assay. This rate was lower than that observed with the synthetic substrate 4-nitrocatechol sulfate, but higher than that seen with the physiological substrate cerebroside sulfate. Extracts of cultured fibroblasts from normal subjects were also shown to hydrolyze ascorbic acid 2-sulfate; extracts of fibroblasts from patients with metachromatic leukodystrophy, known to be deficient in arylsulfatase A, did not. Similarly, hydrolysis of ascorbic acid 2-sulfate was not observed when a partially purified preparation of human arylsulfatase B was tested under a variety of conditions. Thus, in the human, arylsulfatase A appears to be the major, if not the only, ascorbic acid-2-sulfate sulfohydrolase.


Assuntos
Ácido Ascórbico/metabolismo , Cerebrosídeo Sulfatase/metabolismo , Pele/enzimologia , Sulfatases/metabolismo , Células Cultivadas , Fibroblastos/enzimologia , Humanos , Cinética , Relação Estrutura-Atividade
16.
J Invest Dermatol ; 109(3): 370-6, 1997 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9284107

RESUMO

Acne vulgaris is the result of multifactorial disorders of the pilosebaceous duct. The initial lesion is believed to be hyper-keratinization of the infundibulum. The Rhino mouse has been used as an experimental acne model system for screening anti-keratinizing and comedolytic agents. Using this system we show that trypsin could induce desquamation and utriculi-epidermal differentiation in the absence of irritation. Following five daily trypsin treatments, the biomechanical properties of the mouse skin improved, as demonstrated by cutometer measurements and increased elastin expression. Extensive programmed cell death and apoptosis are demonstrated in the utriculi epithelium of the untreated animals. This cell death is eliminated by the trypsin treatment. We speculate that co-administration of trypsin might increase the therapeutic value of topical acne treatments and improve skin elasticity while reducing irritating effects.


Assuntos
Envelhecimento da Pele/efeitos dos fármacos , Pele/citologia , Tripsina/farmacologia , Acne Vulgar/patologia , Animais , Apoptose/efeitos dos fármacos , Cricetinae , Modelos Animais de Doenças , Expressão Gênica/efeitos dos fármacos , Gliceraldeído-3-Fosfato Desidrogenases/genética , Folículo Piloso/efeitos dos fármacos , Ceratolíticos/farmacologia , Masculino , Mesocricetus , Camundongos , Camundongos Pelados , Reação em Cadeia da Polimerase , Precursores de Proteínas/biossíntese , Precursores de Proteínas/genética , RNA Mensageiro/metabolismo , Pele/metabolismo , Pele/patologia
17.
J Invest Dermatol ; 113(2): 272-6, 1999 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10469316

RESUMO

GPIb alpha, a glycoprotein component of the GPIb-IX-V complex, serves as a platelet membrane receptor that mediates adhesion to von Willebrand factor normally present in the vascular subendothelium. Recent data have demonstrated that GPIb alpha is not restricted to platelets, but is also expressed by endothelium in vitro. In this study, we describe the expression and distribution of GPIb alpha in normal adult and neonatal human skin. GPIb alpha is present, as detected by immunohistochemistry, on endothelial cells and on highly dendritic cells localized within the perivascular space, dermal-epidermal junction, and reticular dermis. By dual-labeling immunofluorescence and confocal microscopy, GPIb alpha-positive cells within the dermal interstitium are demonstrated to represent factor XIIIa-positive dermal dendrocytes. In organ cultures of neonatal human foreskin, mast cell degranulation induced by either substance P or compound 48/80 resulted in transiently increased GPIb alpha expression by dermal dendrocytes. Because the GPIb-IX-V complex plays a part in regulating hemostasis and may be important for cellular interactions with extracellular matrix molecules, these data provide additional insight into the potential function of FXIIIa-positive dermal dendrocytes in skin remodeling and repair.


Assuntos
Células Dendríticas/química , Mastócitos/citologia , Glicoproteínas da Membrana de Plaquetas/biossíntese , Glicoproteínas da Membrana de Plaquetas/fisiologia , Receptores de Superfície Celular/biossíntese , Receptores de Superfície Celular/fisiologia , Pele/citologia , Transglutaminases/análise , Adulto , Degranulação Celular , Células Dendríticas/metabolismo , Imunofluorescência , Humanos , Recém-Nascido , Masculino , Técnicas de Cultura de Órgãos , Fenótipo , Complexo Glicoproteico GPIb-IX de Plaquetas/genética , Complexo Glicoproteico GPIb-IX de Plaquetas/farmacocinética , Distribuição Tecidual , Regulação para Cima
18.
J Invest Dermatol ; 115(2): 162-7, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10951231

RESUMO

The chemical basis of melanogenesis is well documented, but the mechanism of melanosome transfer and the regulation of pigmentation by keratinocyte-melanocyte interactions are not well understood. Therefore we examined the effects of serine protease inhibitors on skin pigmentation and found that the protease-activated receptor 2, expressed on keratinocytes, may regulate pigmentation via keratinocyte-melanocyte interactions. Here we show that modulation of protease-activated receptor 2 activation affects melanosome transfer into keratinocytes, resulting in changes in pigment production and deposition. SLIGRL, the protease-activated receptor 2 activating peptide, enhanced melanosome ingestion by keratinocytes, thus increasing pigment deposition. RWJ-50353, a serine protease inhibitor, led to reduced pigment deposition in melanocytes and depigmentation. Electron microscopy studies illustrated an accumulation of immature melanosomes inside melanocytes and abnormal dendrite dynamics in RWJ-50353-treated epidermal equivalents. RWJ-50353 induced a visible and dose-dependent skin lightening effect in the dark-skinned Yucatan swine. Examinations by electron microscopy indicated that the in vivo transfer of melanosomes from melanocytes to keratinocytes was affected. Our data suggest that modulation of keratinocyte-melanocyte interactions via the protease-activated receptor 2 pathway affects melanosome transfer. The use of RWJ-50353 to modulate protease-activated receptor 2 activation could lead to a new class of depigmenting agents.


Assuntos
Melanossomas/fisiologia , Pigmentação da Pele/fisiologia , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Guanidinas/farmacologia , Humanos , Melanossomas/efeitos dos fármacos , Melanossomas/ultraestrutura , Camundongos , Microscopia Eletrônica , Receptor PAR-2 , Receptores de Trombina/fisiologia , Inibidores de Serina Proteinase/farmacologia , Pele/efeitos dos fármacos , Pele/ultraestrutura , Pigmentação da Pele/efeitos dos fármacos , Suínos , Tiazóis/farmacologia
19.
Endocrinology ; 112(3): 862-70, 1983 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-6822216

RESUMO

An organ culture technique for the maintenance of human endometrium was used to study secretory protein production and the ability of progesterone to alter the character of secretory products from this tissue. Proliferative phase cultures of human endometrium were incubated in defined medium for 48 h in the presence or absence of 0.1 microgram/ml progesterone. During the 25th to 48th hours, the tissues were labeled with radioactive protein precursors. The proteins in medium and tissue cytosol were analyzed by one- and two-dimensional polyacrylamide gel electrophoresis, followed by fluorography. Our results suggest that human endometrium secretes at least 24 proteins into the culture medium and that the majority of these are glycoproteins. The presence of progesterone during culture caused a change in the concentration of certain medium proteins. Five protein bands were consistently observed to have a hormonally induced variation of intensity on autoradiographs. Two of these (mol wt, 58,000 and 28,000) showed a decrease in their intensity and three (mol wt, 130,000, 50,000, and 35,000) showed an increase when progesterone-treated and untreated cultures of endometrium were compared. The greatest progesterone-induced change in intensity occurred with the 58,000 mol wt protein (S2). The changes in band intensity appear to be reversible over the time periods studied. Our results provide evidence that significant alterations occur in the protein content of human endometrial secretions as a result of progesterone stimulation.


Assuntos
Endométrio/metabolismo , Progesterona/farmacologia , Proteínas/metabolismo , Adulto , Endométrio/efeitos dos fármacos , Feminino , Humanos , Cinética , Técnicas de Cultura de Órgãos , Precursores de Proteínas/isolamento & purificação , Proteínas/isolamento & purificação
20.
Methods Enzymol ; 190: 334-8, 1990.
Artigo em Inglês | MEDLINE | ID: mdl-2087185

RESUMO

The human sebocyte model offers several advantages over the current animal models. Foremost among these is the correlation of in vitro activity with clinical results, which was not true for arotinoids in the animal models. It is also possible to study several parameters (total cell number, [3H]thymidine uptake, protein and lipid composition/synthesis, hormone response, receptor regulation, etc.) in the same system. The proliferation of isolated sebocytes is inhibited by retinoids, such as isotretinoin and tretinoin, which are known to be clinically active in human acne. Sebocytes are not responsive to the arotinoid temarotene, which is active in the aforementioned animal models and against dimethylbenz[a]anthracene (DMBA)-induced rat mammary carcinoma but inactive clinically in acne. Additionally, this model is not responsive to etretinate, a compound known to be active in psoriasis but inactive in acne. The in vitro model is, therefore, more predicative of clinical efficacy than the animal models alone.


Assuntos
Retinoides/farmacologia , Glândulas Sebáceas/citologia , Divisão Celular/efeitos dos fármacos , Separação Celular/métodos , Células Cultivadas , Técnicas de Cultura/métodos , Etretinato/farmacologia , Humanos , Isotretinoína/farmacologia , Glândulas Sebáceas/efeitos dos fármacos , Tretinoína/farmacologia
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