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1.
Z Gerontol Geriatr ; 52(4): 342-351, 2019 Jul.
Artigo em Alemão | MEDLINE | ID: mdl-29728824

RESUMO

BACKGROUND: Compared with the general population, chronic drug addicts already start showing typical aging problems by the age of 40 years. The increasing number of older drug addicts leads to questions of what an adequate health and social care should look like. This discussion particularly takes place in the context of a sufficient integration of different care systems. A sufficient integration requires an improvement in the networking of substance treatment, nursing care and medical care services. OBJECTIVE: The purpose of this study was to investigate the care structure of older people who use drugs and the services involved in a social network analysis. This was a descriptive design of the pilot study. The study objective was to gain first-hand knowledge about the health and social care situation, the quality of care concerning this client group and to identify supply gaps. Therefore, the three regions Cologne, Dusseldorf and Frankfurt/Main were exemplarily examined. MATERIAL AND METHODS: The data for the social network analysis was gathered by a quantitative online questionnaire. Therefore, especially central network members were contacted and asked to participate. The survey was conducted in two waves. RESULTS: In total, 65 practitioners of all surveyed cities participated in the second wave. The centrality measures assessed indicated that in all regions institutions of the substance abuse service network hold central positions in terms of conveying information. The moderate density values of the networks suggest that there are sufficient cooperation structures. Care deficits were identified most frequently in the areas of housing and nursing care. CONCLUSION: The results provide the first systematic insights and a description of the cooperation practice in the care system. Because of the limitations, further research and practice issues are raised.


Assuntos
Envelhecimento , Usuários de Drogas/psicologia , Equipe de Assistência ao Paciente , Qualidade da Assistência à Saúde , Rede Social , Transtornos Relacionados ao Uso de Substâncias/reabilitação , Idoso , Idoso de 80 Anos ou mais , Cidades , Alemanha , Humanos , Drogas Ilícitas , Comunicação Interdisciplinar , Projetos Piloto , Transtornos Relacionados ao Uso de Substâncias/psicologia
2.
Phys Rev Lett ; 110(22): 225501, 2013 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-23767733

RESUMO

Intermittent or serrated plastic flow is widely observed in the deformation of bulk metallic glasses (BMGs) or other disordered solids at low temperatures. However, the underlying physical process responsible for the phenomena is still poorly understood. Here, we give an interpretation of the serrated flow behavior in BMGs by relating the atomic-scale deformation with the macroscopic shear band behavior. Our theoretical analysis shows that serrated flow in fact arises from an intrinsic dynamic instability of the shear band sliding, which is determined by a critical stiffness parameter in stick-slip dynamics. Based on this, the transition from serrated to nonserrated flow with the strain rate or the temperature is well predicted and the effects of various extrinsic and intrinsic factors on shear band stability can be quantitatively analyzed in BMGs. Our results, which are verified by a series of compression tests on various BMGs, provide key ingredients to fundamentally understand serrated flow and may bridge the gap between the atomic-scale physics and the larger-scale shear band dynamics governing the deformation of BMGs.

3.
Klin Monbl Augenheilkd ; 230(12): 1230-7, 2013 Dec.
Artigo em Alemão | MEDLINE | ID: mdl-24327286

RESUMO

BACKGROUND: Uveal melanoma is the most common primary intraocular malignancy in adults. Iris melanomas are rare tumours: they account for 2-3 % of all uveal melanomas. The clinical differentiation between benign iris nevi and malignant iris melanomas can be difficult. PATIENTS AND METHODS: The aim of this study was the registration, analysis and observation of all patients with tumours of the anterior uvea who had been treated surgically between 1992 and 2011 at the ophthalmic department of the University Hospital of Jena. 40 patients were analysed and compared concerning their preoperative states, operating methods including complications, histological results, postoperative function, subjective complaints as well as the risk of metastasis and the associated dependence on mortality of the dignity of the tumours. In this time period 26 patients has been observed in a follow-up visit. Patients with a malignant tumour were offered an examination. RESULTS: The histological examination revealed for 24 patients a benign tumour and for 16 patients a malignant tumour. After an exact analysis of multiple parameters there was only a statistically significant difference in the preoperative visual acuity (p = 0.025) and in the tumour size (p = 0.011) between the two analysed groups of patients. The rate of serious postoperative complications was 11.4 %. One fourth of the patients complained of subjective problems after the surgical intervention. In the follow-up visit a visual acuity of 0.5 or better was achieved in 68 % of all interventions. CONCLUSIONS: A reliable diagnosis is only possible after histological examination. The analysed parameters can only give indications for the dignity of the tumour. The strategy of the ophthalmic department of the University Hospital of Jena to remove a tumour of uncertain dignity at an early state makes sense, because there are few postoperative complications, few patients complain about subjective problems and the chances for achieving good visual acuity are high.


Assuntos
Iridectomia , Neoplasias da Íris/diagnóstico , Neoplasias da Íris/cirurgia , Melanoma/diagnóstico , Melanoma/cirurgia , Transtornos da Visão/prevenção & controle , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Alemanha , Humanos , Neoplasias da Íris/patologia , Estudos Longitudinais , Masculino , Melanoma/complicações , Melanoma/patologia , Pessoa de Meia-Idade , Resultado do Tratamento , Transtornos da Visão/diagnóstico , Transtornos da Visão/etiologia , Adulto Jovem
4.
Nat Mater ; 9(6): 473-7, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20473286

RESUMO

Bulk metallic glasses (BMGs) generally fail in a brittle manner under uniaxial, quasistatic loading at room temperature. The lack of plastic strain is a consequence of shear softening, a phenomenon that originates from shear-induced dilation that causes plastic strain to be highly localized in shear bands. So far, significant tensile ductility has been reported only for microscopic samples of around 100 nm (ref. 4) as well as for high strain rates, and so far no mechanisms are known, which could lead to work hardening and ductility in quasistatic tension in macroscopic BMG samples. In the present work we developed CuZr-based BMGs, which polymorphically precipitate nanocrystals during tensile deformation and subsequently these nanocrystals undergo twinning. The formation of such structural heterogeneities hampers shear band generation and results in macroscopically detectable plastic strain and work hardening. The precipitation of nanocrystals and their subsequent twinning can be understood in terms of a deformation-induced softening of the instantaneous shear modulus. This unique deformation mechanism is believed to be not just limited to CuZr-based BMGs but also to promote ductility in other BMGs.

5.
Ultrasound Obstet Gynecol ; 38(6): 727-8, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21370302

RESUMO

We report on the pre- and postnatal cytogenetic, molecular genetic and clinical findings in monochorionic-diamniotic twins discordant for trisomy 18. Structural anomalies were identified in one of the twins on prenatal ultrasound examination at 20 weeks' gestation and sampling of amniotic fluid from both sacs was performed for karyotyping. This revealed trisomy 18 in the twin with abnormalities and a normal karyotype in the other twin. Elective Cesarean section was performed at 31 + 5 weeks and the aneuploid twin died shortly after delivery. The surviving twin showed low-grade mosaicism for trisomy 18 on postnatal analysis but has shown normal development. For prenatal diagnosis in monochorionic-diamniotic twin pregnancy the sampling of both amniotic sacs is recommended, especially if one twin has structural anomalies on ultrasound scan.


Assuntos
Amniocentese/métodos , Cromossomos Humanos Par 18/genética , Doenças em Gêmeos/genética , Mosaicismo , Cesárea , Doenças em Gêmeos/diagnóstico , Doenças em Gêmeos/embriologia , Feminino , Humanos , Recém-Nascido , Cariotipagem , Masculino , Mosaicismo/embriologia , Gravidez , Resultado da Gravidez , Diagnóstico Pré-Natal , Gêmeos Monozigóticos/genética
6.
Int J Clin Pharmacol Ther ; 48(1): 1-10, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20040334

RESUMO

OBJECTIVE: N-acetyltransferase 2 (NAT2) genotype-phenotype relation with sulfasalazine as probe drug by means of detailed genotype analysis and kinetic data evaluation. BACKGROUND: Though phenotype analysis of sulfasalazine metabolism has been described before, genotype investigations in this regard are scarce. The influence of different single point mutations on the metabolism of the sulfasalazine metabolite sulfapyridine (SP) should give more insight into the functionality of different alleles especially with those still under discussion. METHODS: In two bioavailability studies performed under comparable conditions with 24 healthy subjects of both genders equally distributed, plasma levels of SP and acetylsulfapyridine (Ac-SP) were determined after oral intake of enteric coated formulations of sulfasalazine (500 mg and 1,000 mg, respectively). The resulting metabolic ratios were calculated. NAT2 genotype was analyzed in parallel for all subjects deducing haplotype set as well as putative functional phenotype as (homozygous or heterozygous) rapid acetylator (RA) or slow acetylator (SA) and correlated with the PK results. RESULTS AND DISCUSSION: RA genotype in the overall study population was seen with 45.5% (including 6.8% homozygous wildtype *4/*4) and SA genotype with 54.5%. Compared to RA genotype, apparent terminal elimination half-life of SP as well as of Ac-SP was prolonged in the SA genotype population, C(max) and AUC values of SP were higher whereas average C(max) value of Ac-SP was lower (with AUC only some tendency to lower values). In general, phenotype-genotype correlation was good with only few exceptions. Strongest functional effect on enzyme activity was noticed in slow acetylators carrying the 341T > C mutation, followed by 590G > A mutation whereas the influence of 857G > A was considerably less pronounced. Homozygous 803A > G mutation (lysine > arginine shift) did not reveal enzyme activity reduction.


Assuntos
Antirreumáticos/farmacocinética , Arilamina N-Acetiltransferase/genética , Sulfassalazina/farmacocinética , Adolescente , Adulto , Antirreumáticos/administração & dosagem , Área Sob a Curva , Disponibilidade Biológica , Ensaios Clínicos como Assunto , Estudos Cross-Over , Relação Dose-Resposta a Droga , Feminino , Genótipo , Meia-Vida , Humanos , Masculino , Pessoa de Meia-Idade , Farmacogenética , Fenótipo , Mutação Puntual , Sulfapiridina/análogos & derivados , Sulfapiridina/farmacocinética , Sulfassalazina/administração & dosagem , Fatores de Tempo , Adulto Jovem
7.
J Gastroenterol Hepatol ; 23(1): 46-50, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18171341

RESUMO

BACKGROUND AND AIM: Although more than 80% of undifferentiated early gastric cancers (EGC) are not associated with lymph node metastasis, endoscopic mucosal resection is not generally accepted as a means of curative treatment because of an abundance of conflicting data concerning clinicopathological characteristics and prognoses. The aim of this study was to define a subgroup of undifferentiated EGC that could be cured by endoscopic treatment without the risk of lymph node metastasis. METHOD: A total of 591 patients surgically resected for undifferentiated EGC between January 1999 and March 2005 were reviewed. Associations between various clinicopathological factors and the presence of lymph node metastasis were analyzed to identify the risk factors of lymph node metastasis. RESULTS: Lymph node metastasis was found in 79 patients (13.4%). By multivariate logistic regression analysis, a tumor diameter 2.5 cm or larger, invasion into the middle third of the submucosal layer or deeper, and lymphatic involvement were identified as independent risk factors of lymph node metastasis (P < 0.001, respectively). Lymph node metastasis was not found in any patient with undifferentiated EGC smaller than 2.5 cm confined to the mucosa or upper third of the submucosal layer without lymphatic involvement. CONCLUSIONS: Undifferentiated intramucosal EGC smaller than 2.5 cm without lymphatic involvement was not associated with lymph node metastasis. Thus, we propose in this circumstance that endoscopic mucosal resection could be considered a definitive treatment without compromising the possibility of cure.


Assuntos
Adenocarcinoma/patologia , Adenocarcinoma/cirurgia , Gastroscopia , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Idoso , Feminino , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco
8.
Plant Biol (Stuttg) ; 10(1): 65-75, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18211548

RESUMO

Methanol emissions from several deciduous tree species with predominantly mature leaves were measured under laboratory and field conditions. The emissions were modulated by temperature and light. Under constant light conditions in the laboratory, methanol emissions increased with leaf temperature, by up to 12% per degree. At constant temperatures, emissions doubled when light intensity (PAR) increased from darkness to 800 micromol x m(-2) x s(-1). A phenomenological description of light and temperature dependencies was derived from the laboratory measurements. This description was successfully applied to reproduce the diel cycle of methanol emissions from an English oak measured in the field. Labelling experiments with (13)CO(2) provided evidence that less than 10% of the emitted methanol was produced de novo by photosynthesis directly prior to emission. Hence, the light dependence of the emissions cannot be explained by instantaneous production from CO(2) fixation. Additional experiments with selective cooling of plant roots indicated that a substantial fraction of the emitted methanol may be produced in the roots or stem and transported to stomata by the transpiration stream. However, the transpiration stream cannot be considered as the main factor that determines methanol emissions by the investigated plants.


Assuntos
Luz , Metanol/metabolismo , Quercus/metabolismo , Quercus/efeitos da radiação , Temperatura , Dióxido de Carbono/metabolismo , Isótopos de Carbono , Ritmo Circadiano/efeitos da radiação , Raízes de Plantas/fisiologia , Raízes de Plantas/efeitos da radiação , Transpiração Vegetal/fisiologia , Transpiração Vegetal/efeitos da radiação
9.
Int J Clin Pharmacol Ther ; 45(1): 36-46, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17256449

RESUMO

OBJECTIVES: The noradrenaline-selective antidepressant reboxetine in vitro is a weak inhibitor of both cytochrome P450 (CYP) 2D6 and CYP3A4. Thus, in this study the pharmacokinetics of reboxetine in relation to pharmacogenetics and the effects of reboxetine compared to paroxetine treatment on the CYP2D6 and CYP3A4 phenotype were analyzed in healthy control subjects. METHODS: Healthy male volunteers were treated with either 6 mg reboxetine (n = 26) or 30 mg paroxetine (n = 25). On Days 10/11 of treatment, serum concentrations of the antidepressants were measured and pharmacokinetic parameters calculated. Volunteers were phenotyped at the end of treatment and after at least 3 weeks washout (true phenotype) using 30 mg dextromethorphan (DM) hydrobromide given orally and measuring DM and metabolites in serum 2 h after intake. CYP2D6 and CYP2C19 genotypes were determined in parallel. RESULTS AND CONCLUSION: Reboxetine serum concentrations showed no correlation with the CYP2D6 genotype and the CYP2D6 phenotype, whereas paroxetine concentrations showed some dependence on CYP2D6. In contrast to in vitro investigations, indicating a major role of CYP3A4 in reboxetine metabolism, reboxetine concentrations in serum showed no correlation with the respective DM metabolic ratios. There was also no correlation between paroxetine concentrations and the CYP3A4 phenotype data. The CYP2C19 genotype (only heterozygosity) had no influence on reboxetine and paroxetine pharmacokinetics. There were only minor changes in the DM metabolite pattern on treatment with reboxetine and no evidence of enzyme inhibition was obtained. In contrast and as expected, paroxetine strongly inhibited CYP2D6. Thus, reboxetine treatment has no effect on the CYP2D6 genotype and no clinically relevant drug interactions involving CYP2D6 are anticipated.


Assuntos
Inibidores da Captação Adrenérgica/farmacocinética , Antidepressivos/farmacocinética , Sistema Enzimático do Citocromo P-450/metabolismo , Morfolinas/farmacocinética , Paroxetina/farmacocinética , Inibidores Seletivos de Recaptação de Serotonina/farmacocinética , Adolescente , Inibidores da Captação Adrenérgica/sangue , Inibidores da Captação Adrenérgica/farmacologia , Adulto , Antidepressivos/sangue , Antidepressivos/farmacologia , Hidrocarboneto de Aril Hidroxilases/metabolismo , Citocromo P-450 CYP2C19 , Citocromo P-450 CYP2D6/metabolismo , Citocromo P-450 CYP3A , Inibidores das Enzimas do Citocromo P-450 , Sistema Enzimático do Citocromo P-450/genética , Dextrometorfano/metabolismo , Interações Medicamentosas , Inibidores Enzimáticos/farmacocinética , Genótipo , Humanos , Masculino , Oxigenases de Função Mista/metabolismo , Morfolinas/sangue , Morfolinas/farmacologia , Mutação , Paroxetina/sangue , Paroxetina/farmacologia , Fenótipo , Reboxetina , Valores de Referência , Inibidores Seletivos de Recaptação de Serotonina/sangue , Inibidores Seletivos de Recaptação de Serotonina/farmacologia
10.
Int J Clin Pharmacol Ther ; 44(5): 225-32, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16724577

RESUMO

OBJECTIVE: The use of cyclosporin A (CSA) and tacrolimus (TAC) in organ transplantation and in the therapy of immune disorders is often hampered by adverse effects, mainly nephro-, hepato- and neurotoxicity. For the development of these side effects, among others, an increased formation of reactive oxygen species, probably generated by the cytochrome P450 (CYP) system, has been accused. Since in this respect literature data are inconsistent, in the present study possible pro- and/or antioxidant effects of CSA and TAC and the involvement of the CYP system were re-evaluated in vitro. METHODS: Effects of CSA and TAC were examined on CYP mediated oxidase functions by stimulated lipid peroxidation (LPO), H2O2 production, and lucigenin (LC) or luminol (LM) amplified chemiluminescence (CL) in liver microsomes of either untreated rats or of rats treated with beta-naphthoflavone (BNF), phenobarbital (PB) or dexamethasone (DEX) and in human liver microsomes. RESULTS: In rat liver microsomes, CSA displayed pro-oxidant properties (though only very slightly), whereas in human liver microsomes small antioxidant effects were seen. With TAC in both species the antioxidant capacity prevailed. Treatment of rats with BNF or DEX caused an increase in the pro-oxidant effects of CSA with respect to LPO or LM-CL, whereas in liver microsomes of DEX-treated rats H2O2 production and LC-CL were diminished. CONCLUSIONS: CSA seems to have both pro-oxidant and antioxidant properties, whereas with TAC mainly an antioxidant capacity was seen. The CYP system seems to be involved in the pro-oxidant influence of CSA. Whether pro-oxidant or antioxidant effects predominate may depend on the antioxidant capacity of a tissue and on the CYP isoforms mainly present.


Assuntos
Antioxidantes/farmacologia , Ciclosporina/farmacologia , Sistema Enzimático do Citocromo P-450/biossíntese , Imunossupressores/farmacologia , Fígado/efeitos dos fármacos , Oxidantes/farmacologia , Tacrolimo/farmacologia , Animais , Dexametasona/farmacologia , Relação Dose-Resposta a Droga , Indução Enzimática , Humanos , Peróxido de Hidrogênio/metabolismo , Técnicas In Vitro , Isoenzimas/biossíntese , Peroxidação de Lipídeos , Fígado/enzimologia , Masculino , Microssomos Hepáticos , Fenobarbital/farmacologia , Ratos , Ratos Wistar , beta-Naftoflavona/farmacologia
11.
Nucleic Acids Res ; 27(19): 3771-8, 1999 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-10481015

RESUMO

The poly(A)-binding protein II (PABP2) is one of the polyadenylation factors required for proper 3'-end formation of mammalian mRNAs. We have cloned Pabp2, the gene encoding the Drosophila homolog of mammalian PABP2, by using a molecular screen to identify new Drosophila proteins with RNP-type RNA-binding domains. Sequence comparison of PABP2 from Drosophila and mammals indicates that the most conserved domains are the RNA-binding domain and a coiled-coil like domain which could be involved in protein-protein interactions. Pabp2 produces four mRNAs which result from utilization of alternative poly(A) sites and encode the same protein. Using an antibody raised against Drosophila PABP2, we show that the protein accumulates in nuclei of all transcriptionally active cells throughout Drosophila development. This is consistent with a general role of PABP2 in mRNA polyadenylation. Analysis of Drosophila PABP2 function in a reconstituted mammalian polyadenylation system shows that the protein has the same functions as its bovine homolog in vitro : it stimulates poly(A) polymerase and is able to control poly(A) tail length.


Assuntos
Drosophila melanogaster/crescimento & desenvolvimento , Proteínas de Insetos/fisiologia , Poli A/metabolismo , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/fisiologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Bovinos , Clonagem Molecular , DNA Complementar , Drosophila melanogaster/genética , Expressão Gênica , Humanos , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Camundongos , Dados de Sequência Molecular , Proteínas de Ligação a Poli(A) , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Coelhos , Homologia de Sequência de Aminoácidos
12.
Matrix Biol ; 24(2): 155-65, 2005 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15890265

RESUMO

The dental follicle is an ectomesenchymal tissue surrounding the developing tooth germ. It is believed that this tissue contains stem cells and lineage committed progenitor cells or precursor cells (PCs) for cementoblasts, periodontal ligament cells, and osteoblasts. In this study, we report the isolation of PCs derived from dental follicle of human third molar teeth. These fibroblast-like, colony forming and plastic adherent cells expressed putative stem cell markers Notch-1 and Nestin. We compared gene expressions of PCs, human mesenchymal stem cells (hMSCs), periodontal ligament cells (PDL-cells) and osteoblasts (MG63) for delimitation of PCs. Interestingly, PCs expressed higher amounts of insulin-like growth factor-2 (IGF-2) transcripts than hMSCs. Differentiation capacity was demonstrated under in vitro conditions for PCs. Long-term cultures with dexamethasone produced compact calcified nodules or appeared as plain membrane structures of different dimensions consisting of a connective tissue like matrix encapsulated by a mesothelium-like cellular structure. PCs differentially express osteocalcin (OCN) and bone sialoprotein (BS) after transplantation in immunocompromised mice but without any sign of cementum or bone formation. Therefore, our results demonstrate that cultured PCs are unique undifferentiated lineage committed cells residing in the periodontium prior or during tooth eruption.


Assuntos
Técnicas de Cultura de Células/métodos , Saco Dentário/patologia , Dente Serotino/citologia , Dente Serotino/patologia , Adolescente , Adulto , Animais , Diferenciação Celular , Linhagem Celular , Linhagem da Célula , Membrana Celular/metabolismo , Primers do DNA/química , Humanos , Imuno-Histoquímica , Imunofenotipagem , Sialoproteína de Ligação à Integrina , Proteínas de Filamentos Intermediários/metabolismo , Mesoderma/citologia , Camundongos , Dente Serotino/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Nestina , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteocalcina/metabolismo , Reação em Cadeia da Polimerase , RNA/metabolismo , RNA Mensageiro/metabolismo , Receptor Notch1 , Receptores de Superfície Celular/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sialoglicoproteínas/metabolismo , Células-Tronco/citologia , Fatores de Tempo , Dente/metabolismo , Fatores de Transcrição/metabolismo
13.
J Mol Biol ; 256(1): 20-30, 1996 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-8609610

RESUMO

Subsets of the four RNA binding domains (RBD 1 to 4) in the Xenopus poly-adenylate binding protein (PABP) have distinct affinities and specificities for RNA. RBDs 1 plus 2 exhibit RNA affinity and selectivity equal to the wild-type (WT) protein. RBDs 3 plus 4 have distinct selectivity and about ten-fold reduced affinity for A23, and the isolated RBDs 2 or 3 or 4 exhibit about 100-fold reduced affinity for A23 in comparison to WT. For the full-length protein, independent RNA contacts have been mapped by UV crosslinking with RBDs 1/2 and RBDs 3/4. The carboxy-terminal, non-RBD portion of the protein does not contribute to RNA affinity or selectivity, but confers homodimerization activity on PABP. RBDs 3 and 4 cooperate with the C terminus to gain poly(A) organizing activity, i.e. the ability to form an RNP with multiple, regularly spaced copies of PABP on a poly(A) substrate.


Assuntos
Proteínas de Ligação a RNA/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Clonagem Molecular , Reagentes de Ligações Cruzadas , DNA/genética , Dados de Sequência Molecular , Estrutura Molecular , Proteínas de Ligação a Poli(A) , Ligação Proteica , RNA/genética , RNA/metabolismo , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Raios Ultravioleta , Xenopus
14.
J Mol Biol ; 297(3): 569-83, 2000 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-10731412

RESUMO

The mammalian nuclear poly(A) binding protein, PABP2, controls the length of the newly synthesized poly(A) tail on messenger RNAs. To gain a better understanding of the mechanism of length control, we have investigated the structure of the PABP2.poly(A) complex. Electron microscopy and scanning force microscopy studies reveal that PABP2, when bound to poly(A), forms both linear filaments and discrete-sized, compact, oligomeric particles. The maximum diameter of the filament is 7 nm; the maximum diameter of the particle is 21(+/-2) nm. Maximum particle size is realized when the PABP2. poly(A) complex is formed with poly(A) molecules 200-300 nt long, which corresponds to the average length of the newly synthesized poly(A) tail in vitro and in vivo. The equilibrium between filaments and particles is highly sensitive to ionic strength; filaments are favored at low ionic strength, while particles predominate at moderate to high ionic strength. Nitrocellulose filter binding and gel mobility shift assays indicate that the PABP2.poly(A) particle formed on A(300) is not significantly more stable than complexes formed with smaller species of poly(A). These results are discussed in the context of the proposed functions for PABP2.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a DNA/ultraestrutura , Poli A/metabolismo , Proteínas de Ligação a RNA/metabolismo , RNA/metabolismo , RNA/ultraestrutura , Regulação Alostérica , Animais , Ligação Competitiva , Biopolímeros/química , Biopolímeros/metabolismo , Bovinos , Proteínas de Ligação a DNA/química , Concentração de Íons de Hidrogênio , Microscopia de Força Atômica , Microscopia Eletrônica , Peso Molecular , Concentração Osmolar , Poli A/síntese química , Poli A/química , Poli A/genética , Proteína II de Ligação a Poli(A) , Proteínas de Ligação a Poli(A) , Ligação Proteica , RNA/química , RNA/genética , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/ultraestrutura , Termodinâmica
15.
J Immunol Methods ; 179(2): 215-22, 1995 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-7876569

RESUMO

The incorporation of radioactive nucleotides into newly synthesized DNA has been established as a standard method for the detection of proliferation in eucaryotic cells. Unfortunately the use of this method makes it harder to obtain information on the phenotype of proliferating cells in mixed cell populations. For this reason we established a flow-cytometric approach employing a monoclonal antibody specific for murine as well as human proliferating cell nuclear antigen (PCNA) and a double labeling technique for detection of cell membrane-expressed phenotypic markers. The efficiency of this immunostaining procedure was confirmed by simultaneous and highly specific detection of PCNA in nuclear structures as well as cell membrane-expressed antigens using cytological techniques. In vitro experiments with mitogen- and alloantigen-stimulated murine lymph node cells (LNC) and human peripheral blood mononuclear leukocytes (PBML) revealed a good correlation of total [3H]thymidine incorporation into DNA and expression of PCNA. For the analysis of proliferating cells activated in vivo the method was employed to evaluate the local lymph node assay which assesses the allergenicity of small chemicals. LNC prepared from the cervical lymph nodes of mice treated on 4 consecutive days with sensitizing concentrations of the contact allergens oxazolone, TNCB and DNFB as well as the irritants benzoic acid and SLS in comparison to the solvent control showed a dramatic increase in the total amount of proliferating cells for contact allergen-treated animals in comparison to the solvent control and irritant-treated mice. In addition a detailed phenotyping of the proliferating cell populations was possible. This approach offers an easy to perform, non-radioactive method for the assessment of proliferation of murine as well as human leukocytes in vitro and especially in vivo and will be of great advantage for situations where the phenotype of proliferating cellular subsets in heterogeneous populations is of interest.


Assuntos
Antígeno Nuclear de Célula em Proliferação/análise , Alérgenos/farmacologia , Animais , Feminino , Citometria de Fluxo , Imunofenotipagem , Técnicas In Vitro , Linfonodos/citologia , Ativação Linfocitária , Subpopulações de Linfócitos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL
16.
J Immunol Methods ; 203(2): 171-80, 1997 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-9149811

RESUMO

The aim of this study was to explore the usefulness of human blood dendritic cells (DC) in the development of an in vitro model for predictive testing of contact sensitizers. A method was established to monitor the influence of chemicals on the intracellular targeting of antibody-crosslinked MHC class II molecules after their uptake by human DC. Using a three-colour flow-cytometric technique, freshly prepared DC were distinguished from other MHC class II-bearing cell types such as B-cells and monocytes in unseparated mononuclear cell suspensions of healthy volunteers. The assay is based on the pH-sensitivity of internalized fluorescein-coupled MHC class II specific antibodies. Quenching of fluorescence intensity due to internalization into acidic intracellular compartments was observed with untreated DC whereas internalization into less acidic structures following stimulation with strong contact sensitizers ensured that the fluorescence intensity was conserved. The usefulness of this approach for predictive testing of the preservatives MI/MCI, imidazolidinyl urea, methyl-4-hydroxy-benzoate and 2-phenoxyethanol in comparison to the strong allergen DNFB and the irritants sodium lauryl sulphate and dithranol was explored. Whereas low concentrations of MI/MCI resembled the strong allergen DNFB, high concentrations of imidazolidinyl urea were required for a moderate response. Methyl-4-hydroxy-benzoate and 2-phenoxyethanol as well as the irritants SLS and dithranol failed to induce a significant effect in this assay. The non-responsiveness to the latter compounds reflected their minor or absent capacity to induce contact hypersensitivity in humans, whereas DNFB, MI/MCI and imidazolidinyl urea are well established contact sensitizers. These data suggest that the capacity of a chemical to modulate endocytotic mechanisms in dendritic cells in vitro seems to reflect the probability of that substance acting as a hapten in vivo.


Assuntos
Células Dendríticas/metabolismo , Endocitose/imunologia , Citometria de Fluxo/métodos , Haptenos/farmacologia , Receptores Imunológicos/fisiologia , Anticorpos Monoclonais/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Endocitose/efeitos dos fármacos , Fluoresceínas/metabolismo , Fluorescência , Antígenos HLA-DR/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Líquido Intracelular/efeitos dos fármacos , Líquido Intracelular/imunologia , Líquido Intracelular/metabolismo , Valor Preditivo dos Testes , Receptores Imunológicos/efeitos dos fármacos
17.
Immunol Lett ; 26(2): 115-9, 1990 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-2269481

RESUMO

In order to remove the EAI "blocking" activity, EA inhibition positive sera from patients with recurrent abortions were absorbed after alloimmunization with paternal lymphocytes by the lymphoblastoid cell line (LCL) of the husband. To avoid non-specific binding via the Fc-receptor, the cells were first fixed with 0.05% glutaraldehyde. Absorption was performed for 3 h at 4 degrees C. For subsequent elution, the cells were incubated with 0.1 M glycine-HCl, pH 2.3, for 30 min at 4 degrees C. After each step, EAI assay was carried out to determine the "blocking" activity in the supernatants. Furthermore, 10% SDS-PAGE and immunoblotting with goat antihuman IgG of the whole serum and the supernatants were performed. The results obtained give evidence that the EAI "blocking" activity can be absorbed by and eluted from the LCL of the immunizing husband, and is due to an IgG antibody directed against an antigen present on the LCL. Further absorption experiments with trophoblast cells will show whether this antigen is also displayed by the trophoblast.


Assuntos
Aborto Habitual/terapia , Fatores Biológicos/imunologia , Imunização , Isoantígenos/administração & dosagem , Aborto Habitual/imunologia , Linhagem Celular Transformada , Citotoxicidade Imunológica/imunologia , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Immunoblotting , Linfócitos/imunologia , Masculino , Gravidez , Receptores Fc/imunologia , Formação de Roseta
18.
J Reprod Immunol ; 20(2): 141-51, 1991 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-1836500

RESUMO

In a prospective study 140 couples who had at least three spontaneous abortions (RSA) were studied for the presence of Fc receptor blocking antibodies detected by the erythrocyte antibody rosette inhibition (EAI) assay, for anti-paternal cytotoxic antibodies (APCA), and for mixed lymphocyte culture inhibiting (MLCI) antibodies before and after active immunization with paternal lymphocytes. The comparative analysis revealed the EAI assay to possess a higher sensitivity than the APCA and/or MLCI tests in monitoring the specific immune response after active immunization. The success of pregnancy in EAI positive post-immunization patients was not influenced by the presence or absence of APCA or MLCI. In the light of a successful pregnancy outcome of 85.7% (n = 37) in this study we conclude that the monitoring of Fc receptor blocking antibodies is useful in active immunization protocols for RSA patients.


Assuntos
Aborto Habitual/imunologia , Gravidez/imunologia , Receptores Fc/imunologia , Aborto Habitual/terapia , Autoimunidade , Linfócitos B/imunologia , Testes Imunológicos de Citotoxicidade , Feminino , Humanos , Imunização Passiva , Imunoterapia , Teste de Cultura Mista de Linfócitos , Monócitos/imunologia , Estudos Prospectivos , Formação de Roseta , Linfócitos T/imunologia
19.
J Reprod Immunol ; 15(2): 159-67, 1989 May.
Artigo em Inglês | MEDLINE | ID: mdl-2769649

RESUMO

To analyse the nature of antibodies which are purported to be essential for the maintenance of normal human pregnancy, six centers participated in a workshop of "blind" tests on 19 allosera. Fc-receptor dependent assays detected antibodies with specificity only for HLA. In addition to cytotoxic antibodies, the Fc-receptor dependent immune phagocytosis inhibition test revealed two non-cytotoxic alloantibodies with HLA specificity. These antibodies had high titers and may, therefore, be essentially non-cytotoxic. Murine monoclonal antibodies to HLA-A, B, C or DR (W6/32 and 2MC3) were used to evaluate the methods. These antibodies inhibited immune rosette formation as well as immune phagocytosis. Diluted to concentrations below the threshold of complement-dependent cytotoxicity, the monoclonal antibodies still inhibited the mixed lymphocyte reaction and the immune phagocytosis. A human monoclonal immunoglobulin M with specificity for monomorphic non-HLA lymphocyte antigens inhibited the mixed lymphocyte reaction. The immune rosette inhibition test exhibited several false positive reactions, e.g. three out of four with a serum that did not contain alloantibodies to blood cells. Non-cytotoxic antibodies were therefore rare in the selected sera of the workshop and they exhibited HLA specificity only. No participant was able to identify pregnancy-maintaining non-HLA-antibodies.


Assuntos
Aborto Habitual/terapia , Isoanticorpos/análise , Isoantígenos/uso terapêutico , Gravidez/imunologia , Citotoxicidade Imunológica , Feminino , Antígenos HLA/imunologia , Humanos , Imunização , Isoantígenos/imunologia , Linfócitos/imunologia , Masculino , Fagocitose , Formação de Roseta
20.
Science ; 183(4130): 1142, 1974 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-17789206
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