RESUMO
Both weekly cisplatin chemotherapy and single agent topotecan have proven to be effective in recurrent ovarian cancer. Preclinical data show synergism between cisplatin and topotecan. Side effects for this combination are drug sequence dependent and predominantly haematologic. Since preclinical data suggest that Cremophor EL (CrEL), the formulation vehicle of paclitaxel, has a protective effect on haematological toxicity of cisplatin, CrEL was added to the combination cisplatin and topotecan. In this phase I study, escalating doses of oral topotecan administered on day 1, 2, 8, 9, 15, 16, 29, 30, 36, 37, 43, 44 were combined with weekly cisplatin 70 mg m(-2) d(-1) on day 1, 8, 15, 29, 36, 43 (scheme A) or with the presumably less myelotoxic sequence weekly cisplatin day 2, 9, 16, 30, 37, 44 (scheme B). In scheme C, CrEL 12 ml was administered prior to cisplatin in the sequence of Scheme A. 18 patients have received a total of 85 courses. In scheme A 4/10 patients, all treated with topotecan 0.45 mg m(-2) d(-1), experienced DLT: 1 patient had vomiting grade 4, 1 patient had grade 4 neutropenia >5 days, 1 patient had >2 weeks delay due to thrombocytopenia and 1 patient due to neutropenia. Both patients in scheme B (topotecan 0.45 mg m(-2) d(-1)) had DLT due to a delay > 2 weeks because of prolonged haematological toxicity. No DLT was observed in the first 3 patients in scheme C (topotecan 0.45 mg m(-2) d(-1)). However, 2 out of 3 patients treated at dose level topotecan 0.60 mg m(-2)d(-1) in scheme C experienced DLT due to >2 weeks delay because of persistent thrombocytopenia or neutropenia. We conclude that there is a modest clinical effect of CrEL on haematological toxicity for this cisplatin-based combination regimen, which seems to reduce these side effects but does not really enable an increase of the oral topotecan dose.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Cisplatino/administração & dosagem , Glicerol/análogos & derivados , Recidiva Local de Neoplasia/tratamento farmacológico , Neoplasias Ovarianas/tratamento farmacológico , Topotecan/administração & dosagem , Administração Oral , Adulto , Idoso , Cisplatino/farmacocinética , Esquema de Medicação , Feminino , Glicerol/administração & dosagem , Humanos , Pessoa de Meia-Idade , Topotecan/farmacocinéticaRESUMO
We compared intraindividual and interindividual variability in the plasma levels of fibrinogen, tissue-type plasminogen activator (TPA) antigen, plasminogen activator inhibitor (PAI) activity, and C-reactive protein (CRP) in 20 healthy, young individuals and 26 patients with stable angina pectoris (AP) who were at higher risk for cardiovascular disease. For each of the four parameters, the contribution of the intraindividual variation to the total variance (13% and 9% for fibrinogen, 3% and 5% for TPA antigen, 4% and 20% for In[PAI activity], and 14% and 9% for In[CRP] for the healthy volunteers and AP patients, respectively) was smaller than the contribution from the interindividual variation. These results indicate that single sampling is sufficient to assess an individual level for TPA antigen and PAI activity, whereas duplicate sampling for fibrinogen and triplicate sampling for CRP are recommended. In an epidemiological study the sample sizes, based on the variances found in the transverse part of the study, needed to detect a 15% difference between the two groups (with alpha = 0.01 and a statistical power = .90) are 31 and 40 for fibrinogen, 568 and 146 for TPA antigen, 603 and 119 for PAI activity, and 1490 and 2263 for CRP in healthy volunteers and patients with AP, respectively. Additionally, we studied the contribution of genetic polymorphisms of the B beta-fibrinogen (Bcl I and G-->A-455) and PAI activity (HindIII and CA-repeat) genes to intraindividual and interindividual variation. Fibrinogen genotypes were associated with plasma fibrinogen levels in the volunteers but not in the AP patients. No effects of fibrinogen or PAI polymorphisms on intraindividual variation were observed in either healthy individuals or AP patients. In this study intraindividual variation in plasma levels of the cardiovascular risk indicators fibrinogen, TPA antigen, PAI activity, and CRP was small when compared with the interindividual variation in healthy, young volunteers and patients with stable AP.
Assuntos
Angina Pectoris/sangue , Proteína C-Reativa/análise , Fibrinogênio/análise , Inativadores de Plasminogênio/análise , Ativador de Plasminogênio Tecidual/análise , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Pessoa de Meia-IdadeRESUMO
Two forms of histidine-rich glycoprotein (HRG) were detected on SDS-PAGE by silver staining and immunoblotting after isolation of the protein from pooled plasma using immuno-affinity chromatography followed by chromatography with heparin-Sepharose. Both forms were single-chain molecules and the apparent molecular weights of form 1 and form 2 were 77 kD and 75 kD respectively. Mendelian inheritance of both HRG forms was observed in four families with 24 informative meioses, strongly suggesting that the two forms are encoded by different alleles. The frequency of form 1 and form 2 in a group of 36 individuals was 0.35 and 0.65 respectively. The difference between the two molecular variants was studied by direct sequence analysis of amplified exons of the HRG gene from 6 individuals who were homozygous either for form 1 or form 2. Five amino acid polymorphisms in three different exons were observed: Ile/Thr in exon4; Pro/Ser in exon 5; His/Arg, Arg/Cys and Asn/Ile in exon 7. Analysis of these polymorphisms in 20 volunteers showed that only the Pro/Ser polymorphism at position 186 in exon 5 was coupled to the form of the HRG protein. Ser was found in form 1 and Pro in form 2. The presence of Ser at position 186 introduces a consensus sequence for a N-glycosylation site (Asn-X-Ser/Thr). By removing N-linked sugars with N-glycanase, it could be demonstrated that the difference between the two forms of HRG is caused by an extra carbohydrate group at Asn 184 in form 1.
Assuntos
Proteínas Sanguíneas/genética , Variação Genética , Glicoproteínas/genética , Polimorfismo Genético , Proteínas/genética , Sequência de Aminoácidos , Sequência de Bases , Estudos de Casos e Controles , Éxons , Código Genético , Genótipo , Humanos , Dados de Sequência Molecular , Peso Molecular , Linhagem , FenótipoRESUMO
A pedigree-based maximum likelihood method developed by Lange et al. (12) was used to study the contribution of a newly defined di-allelic polymorphism in histidine-rich glycoprotein (HRG) to the plasma levels of HRG. In four families (n = 99) and 20 volunteers we found a heritability of 70%, an age effect of 3% and an effect of individual environmental factors of 27%. These results are remarkably similar to the results found in a previous parent-twin study in which a heritability of 69% and an effect of random environment of 31% was found. The overall genetic influence in the present study can be subdivided into an effect of 59% by the HRG phenotype and 11% by residual genetic factors. The influence of the HRG phenotype of 59% can entirely be explained by adding up the effect of the two alleles that make up the phenotype. These results indicate a codominant inheritance pattern of HRG levels in which the genetic influences can almost completely be ascribed to the additive effect of the di-allelic HRG locus whereas only a small part is due to other loci.
Assuntos
Aminoácidos/química , Proteínas Sanguíneas/genética , Glicoproteínas/genética , Polimorfismo Genético , Proteínas/genética , Alelos , Análise de Variância , Proteínas Sanguíneas/metabolismo , Estudos de Casos e Controles , Mapeamento Cromossômico , Glicoproteínas/sangue , Humanos , Funções Verossimilhança , Peso Molecular , Linhagem , Fenótipo , Proteínas/metabolismoRESUMO
Recent studies describe families with both elevated plasma HRG levels and thrombosis. In order to study the possibility that allelic variants of the HRG locus are associated with differences in HRG level, we studied linkage between HRG levels and a dinucleotide repeat polymorphism in a Dutch family which was selected on the presence of both thrombosis and elevated plasma HRG levels. No other known risk factors from thrombosis were found in this family. Linkage was calculated between the dinucleotide repeat and the HRG level considering the HRG level as a quantitative phenotype assuming a population prevalence of elevated HRG of 5%. Two classes of HRG levels were defined by a mean and a variance: one class with normal HRG levels and a second class with high HRG levels. Using a mean HRG level of 99% for individuals with a normal HRG level and 145% for individuals with high HRG, a maximum lod score of 4.17 (odds in favour of linkage of 22,000:1) was found at a recombination fraction of 0, indicating linkage. Considering the pedigree, an association was found between the presence of a specific allele (no. 6) of the dinucleotide repeat polymorphism and plasma HRG levels. Family members carrying allele 6 were found to have higher HRG plasma levels compared with family members lacking allele 6 (149% v 109% respectively). We conclude that in this family, linkage is found between the HRG locus and the HRG level, and that a HRG gene coupled to allele 6 of the dinucleotide polymorphism is associated with elevated plasma HRG levels. No evidence was found for a causal relationship between elevated plasma HRG levels and thrombosis in this family.
Assuntos
Alelos , Glicoproteínas/sangue , Proteínas/metabolismo , Trombose/genética , Idoso , Sequência de Bases , Coagulação Sanguínea , Feminino , Ligação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Linhagem , Polimorfismo Genético , Sequências Repetitivas de Ácido Nucleico , Trombose/sangueRESUMO
Plasma levels of histidine-rich glycoprotein (HRG) were investigated in three groups of women receiving a different dose of estrogens. First, the effect of low-dose estrogen was studied in a group of 83 postmenopausal women who were treated with 0.625 mg conjugated estrogens (CE). No significant change from baseline levels was found at the end of cycle 3 and cycle 13. Secondly, in 15 mothers and 23 daughters using oral contraceptives (OC) containing 30-50 micrograms ethinyl estradiol (EE) daily the mean HRG level was 14% and 24% lower than in a group of 144 mothers and 134 daughters not taking oral contraceptives, respectively (p < 0.05). Finally, in 11 excessively tall prepuberal girls who received 300 micrograms EE daily to reduce their final height the mean plasma HRG levels were decreased by 68% (p < 0.005). The effect of progestogens administered during low-dose and high-dose estrogen therapy appeared to be minor. The results from these three studies indicate that estrogens reduce plasma HRG levels in a dose-dependent way.
Assuntos
Estrogênios Conjugados (USP)/farmacologia , Etinilestradiol/farmacologia , Medrogestona/farmacologia , Acetato de Medroxiprogesterona/farmacologia , Proteínas/análise , Adolescente , Fatores Etários , Idoso , Estatura/efeitos dos fármacos , Criança , Anticoncepcionais Orais Hormonais/efeitos adversos , Anticoncepcionais Orais Hormonais/farmacologia , Relação Dose-Resposta a Droga , Estrogênios Conjugados (USP)/efeitos adversos , Etinilestradiol/efeitos adversos , Feminino , Gigantismo/prevenção & controle , Humanos , Pessoa de Meia-Idade , Pós-Menopausa , Risco , Tromboembolia/sangue , Tromboembolia/induzido quimicamente , Tromboembolia/epidemiologiaAssuntos
Cromossomos Humanos Par 3 , Genes , Íntrons , Proteínas/genética , Deleção de Sequência , Sequência de Bases , Linhagem Celular , Mapeamento Cromossômico , Cosmídeos , Cistatinas/genética , Humanos , Hibridização in Situ Fluorescente , Cininogênios/genética , Dados de Sequência Molecular , Polimorfismo Genético , Sequências Repetitivas de Ácido Nucleico , Homologia de Sequência do Ácido NucleicoRESUMO
Plasma histidine-rich glycoprotein (HRG) was found to be persistently increased in a patient with a history of recurrent arterial thromboembolic events. The mean concentration was 270% of normal pooled plasma. Increased HRG was found in eight of the 17 relatives studied, but none of them has experienced thrombo-embolism yet. Apparently, increased HRG was hereditary with autosomal dominant inheritance. A significant correlation was found between the increased plasma concentration of the protein and the age of the subjects (P < 0.02), whereas no such relation is present in a normal population. The plasma HRG of the proposita and 9 of her family members displayed abnormal binding to heparin, as assessed in a crossed affinity immuno-electrophoresis system: the usual increase in mobility after binding to heparin was absent. The binding of this variant HRG to plasminogen was normal. This case represents the first abnormal HRG variant reported and it is proposed to designate it: HRG Eindhoven.
Assuntos
Proteínas Sanguíneas/metabolismo , Glicoproteínas/sangue , Heparina/sangue , Proteínas/metabolismo , Feminino , Variação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Linhagem , Estudos Prospectivos , Ligação ProteicaRESUMO
Histidine-rich glycoprotein (HRG) is a non-enzymatic glycoprotein that acts as a modulator of several plasma proteins involved in coagulation and fibrinolysis. The contributions of genetic and environmental influences to inter-individual variation in plasma levels of HRG were studied in 160 Dutch families consisting of adolescent twin pairs and their parents. Results showed that 69% of the variance in plasma HRG concentrations could be accounted for by genetic factors. Heritability was the same in males and females and in parents and their offspring. There was no association between HRG levels of husband and wife and no evidence was found for the influence of shared family environment on the resemblance between relatives.
Assuntos
Pais , Proteínas/análise , Gêmeos , Adolescente , Adulto , Fatores Etários , Meio Ambiente , Feminino , Genótipo , Humanos , Funções Verossimilhança , Masculino , Pessoa de Meia-Idade , Proteínas/genética , Fatores Sexuais , Gêmeos Dizigóticos , Gêmeos MonozigóticosRESUMO
The estrogen-dependent binding of a protein to the upstream region of the chicken vitellogenin gene was detected by using in vivo dimethyl sulfate, genomic DNase I, and in vitro exonuclease III footprinting. The site is located between base pairs -848 and -824, and its sequence resembles that of the nuclear factor I binding site. The results suggest that a nuclear factor binding to this site is involved in the regulation of the vitellogenin gene.
Assuntos
Proteínas de Ligação a DNA/fisiologia , Estrogênios/fisiologia , Regulação da Expressão Gênica , Proteínas Nucleares/fisiologia , Sequências Reguladoras de Ácido Nucleico , Vitelogeninas/genética , Animais , Sequência de Bases , Sítios de Ligação , Galinhas , Dados de Sequência MolecularRESUMO
Protein-DNA interactions in the promoter region of the chicken vitellogenin II gene were analyzed by in vivo dimethylsulphate footprinting with expressing and non-expressing tissues. The reactivity of G-residues is essentially the same in erythrocytes, oviduct and control liver, not expressing the gene. In the expressing estrogen-induced liver we find a number of G-residues with altered reactivities. These G's are located within distinct sequences: the estrogen responsive elements, a sequence resembling the NF-1 recognition motive, and several elements which are conserved between yolk protein genes. The expression-dependent binding of proteins to these sites was confirmed by DNaseI footprinting applied to nuclei isolated from estrogen-induced and control liver. Estradiol appears to establish a transcription complex comprising a number of distinct proteins bound to different sites in the 5' flanking region of the vitellogenin II gene.