RESUMO
Surgical results of 76 patients (76 eyes) aged 25-79 years with secondary refractory glaucoma were analyzed. The best hypotensive effect and visual functions integrity were achieved with Ahmed valve implantation (86.7% and 83.3% of cases respectively); after conventional fistulizing surgery the hypotensive effect was observed in 45.5%, noncompromised vision--in 54.5% of cases. Tunnel trabeculectomy with iridocycloretraction led to normalization of intraocular pressure and stabilization of visual functions in 81.3% and 68.8% of cases respectively and thus can be considered as an alternative to fistulizing surgery in patients with secondary refractory glaucoma. Uveal glaucoma is a relative contraindication to Ahmed valve implantation, while neovascular glaucoma is that to tunnel trabeculectomy with iridocycloretraction.
Assuntos
Implantes para Drenagem de Glaucoma , Glaucoma/etiologia , Glaucoma/cirurgia , Adulto , Idoso , Feminino , Glaucoma/fisiopatologia , Humanos , Pressão Intraocular , Masculino , Pessoa de Meia-Idade , Tonometria Ocular , Trabeculectomia , Resultado do Tratamento , Acuidade VisualRESUMO
The paper presents the results of studying the effect of 10% perfluorane (PF) emulsion intravenously injected 1-2 days before surgical treatment for rhegmatogenous retinal detachment on the levels of the cytokines IL-1beta, TNF-alpha, IL-6, and IL-4 in the serum and subretinal fluid of patients. PF infusion was found to exert an immunomodulatory effect that favored a short-term increase in the serum level of proinflammatory cytokines (IL- 1beta and IL-6) at week 1 and TNF-alpha on day 1) with their gradual normalization. The subretinal fluid showed recovery of the mechanisms for local immunological reactions; eye inflammation reduced due to the decreased production of proinflammatory cytokines and simultaneously the elevated level of the anti-inflammatory cytokine IL-4.
Assuntos
Fluorocarbonos/administração & dosagem , Imunomodulação/efeitos dos fármacos , Cuidados Pré-Operatórios/métodos , Descolamento Retiniano/imunologia , Adulto , Substitutos Sanguíneos/administração & dosagem , Citocinas/metabolismo , Feminino , Seguimentos , Humanos , Injeções Intravenosas , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Oftalmológicos/métodos , Prognóstico , Descolamento Retiniano/metabolismo , Descolamento Retiniano/cirurgia , Líquido Sub-Retiniano/metabolismoRESUMO
The paper deals with complex treatment for pleural malignancies with concomitant effusions. Cytoreduction and abatement of effusion, protein loss, inflammation, intoxication and pain syndrome were reported after argon-plasma electrocoagulation of the pleura followed by photodynamic therapy and hyperthermal intrapleural chemoperfusion. Stable effusion abatement effect was confirmed in all eight cases by X-ray examination and changes in homeostatic indices and breathing function as well as lowered severity of intoxication and pain. No emergency repeat intervention or pleural puncture was reported.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Quimioterapia do Câncer por Perfusão Regional , Eletrocoagulação , Fotoquimioterapia , Derrame Pleural/complicações , Neoplasias Pleurais/complicações , Neoplasias Pleurais/terapia , Pleurisia/complicações , Adulto , Idoso , Quimioterapia do Câncer por Perfusão Regional/métodos , Feminino , Humanos , Hipertermia Induzida , Masculino , Pessoa de Meia-Idade , Neoplasias Pleurais/tratamento farmacológico , Neoplasias Pleurais/cirurgia , Resultado do TratamentoRESUMO
Mutations in the cytoplasmic domain of the insulin receptor that block the ability of the receptor to stimulate glucose uptake do not block the receptor's ability to inhibit apoptosis (Boehm, J. E., Chaika, O. V., and Lewis, R. E. (1998) J. Biol. Chem. 273, 7169-7176). To characterize this survival pathway we used a chimeric receptor (CSF1R/IR) consisting of the ligand-binding domain of the colony-stimulating factor-1 receptor spliced to the cytoplasmic domain of the insulin receptor and a mutated version of the chimeric receptor containing a 12-amino acid deletion of the juxtamembrane domain (CSF1R/IRDelta960). In addition to the inhibition of apoptosis, activation of either the CSF1R/IR or the CSF1R/IRDelta960 rapidly induced membrane ruffling in Rat1 fibroblasts. The small GTPase Rac mediates membrane ruffling. Activated and dominant-inhibitory mutants of Rac and other small GTPases were expressed in Rat1 fibroblasts to examine a potential link between the intracellular pathways that induce membrane ruffling and promote cell survival. The anti-apoptotic action of the CSF1R/IRDelta960 was reversed by dominant-inhibitory Rac(N17), but not by Ras(N17) or Cdc42(N17). Activated Rac(V12), but not Ras(D12) or Cdc42(V12), promoted cell survival in the absence of insulin. These data implicate Rac as a mediator of an unique anti-apoptotic signaling pathway activated by the insulin receptor cytoplasmic domain.
Assuntos
Apoptose , Citoplasma/metabolismo , Receptor de Insulina/metabolismo , Transdução de Sinais , Proteínas rac de Ligação ao GTP/metabolismo , Animais , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Insulina/farmacologia , Fator Estimulador de Colônias de Macrófagos/farmacologia , Ratos , Proteínas Recombinantes/metabolismoRESUMO
The kinase suppressor of Ras (KSR) is a loss-of-function allele that suppresses the rough eye phenotype of activated Ras in Drosophila and the multivulval phenotype of activated Ras in Caenorhabditis elegans. The physiological role of mammalian KSR is not known. We examined the mechanisms regulating the phosphorylation of this putative kinase in mammalian cells. Wild-type mouse KSR and a mutated KSR protein predicted to create a kinase-dead protein are phosphorylated identically in intact cells and in the immune complex. Phosphopeptide sequencing identified 10 in vivo phosphorylation sites in KSR, all of which reside in the 539 noncatalytic amino terminal amino acids. Expression of the amino terminal portion of KSR alone demonstrated that it was phosphorylated in the intact cell and in an immune complex in a manner indistinguishable from that of intact KSR. These data demonstrate that the kinase domain of KSR is irrelevant to its phosphorylation state and suggest that the phosphorylation of KSR and its association with a distinct set of kinases may affect intracellular signaling.
Assuntos
Proteínas Quinases/metabolismo , Proteínas ras/metabolismo , Sequência de Aminoácidos , Animais , Complexo Antígeno-Anticorpo/genética , Complexo Antígeno-Anticorpo/metabolismo , Sítios de Ligação/genética , Linhagem Celular , Embrião de Mamíferos , Humanos , Rim/citologia , Camundongos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Fosforilação , Testes de Precipitina , Desnaturação Proteica , Proteínas Quinases/química , Proteínas Quinases/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Deleção de Sequência , Serina/genética , Proteínas ras/antagonistas & inibidoresRESUMO
CSF-1 is equipotent to insulin in its ability to stimulate 2-[3H]deoxyglucose uptake in 3T3-L1 adipocytes expressing the colony stimulating factor-1 receptor/insulin receptor chimera (CSF1R/IR). However, CSF-1-stimulated glucose uptake and glycogen synthesis is reduced by 50% in comparison to insulin in 3T3-L1 cells expressing a CSF1R/IR mutated at Tyr960 (CSF1R/IRA960). CSF-1-treated adipocytes expressing the CSF1R/IRA960 were impaired in their ability to phosphorylate insulin receptor substrate 1 (IRS-1) but not in their ability to phosphorylate IRS-2. Immunoprecipitation of IRS proteins followed by Western blotting revealed that the intact CSF1R/IR co-precipitates with IRS-2 from CSF-1-treated cells. In contrast, the CSF1R/IRA960 co-precipitates poorly with IRS-2. These observations suggest that Tyr960 is important for interaction of the insulin receptor cytoplasmic domain with IRS-2, but it is not essential to the ability of the insulin receptor tyrosine kinase to use IRS-2 as a substrate. These observations also suggest that in 3T3-L1 adipocytes, tyrosine phosphorylation of IRS-2 by the insulin receptor tyrosine kinase is not sufficient for maximal stimulation of receptor-regulated glucose transport or glycogen synthesis.
Assuntos
Adipócitos/metabolismo , Glucose/metabolismo , Fosfoproteínas/metabolismo , Receptor de Insulina/metabolismo , Tirosina/metabolismo , Células 3T3 , Animais , Transporte Biológico , Membrana Celular/metabolismo , Citoplasma/metabolismo , Ativação Enzimática , Proteínas Substratos do Receptor de Insulina , Peptídeos e Proteínas de Sinalização Intracelular , Ligantes , Fator Estimulador de Colônias de Macrófagos/farmacologia , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Receptor de Insulina/genética , Receptor de Fator Estimulador de Colônias de Macrófagos/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Tirosina/genéticaRESUMO
Kinase suppressor of Ras (KSR) is a loss-of-function allele that suppresses the rough eye phenotype of activated Ras in Drosophila and the multivulval phenotype of activated Ras in Caenorhabditis elegans. Genetic and biochemical studies suggest that KSR is a positive regulator of Ras signaling that functions between Ras and Raf or in a pathway parallel to Raf. We examined the effect of mammalian KSR expression on the activation of extracellular ligand-regulated (ERK) mitogen-activated protein (MAP) kinase in fibroblasts. Ectopic expression of KSR inhibited the activation of ERK MAP kinase by insulin, phorbol ester, or activated alleles of Ras, Raf, and mitogen and extracellular-regulated kinase. Expression of deletion mutants of KSR demonstrated that the KSR kinase domain was necessary and sufficient for the inhibitory effect of KSR on ERK MAP kinase activity. KSR inhibited cell transformation by activated RasVal-12 but had no effect on the ability of RasVal-12 to induce membrane ruffling. These data indicate that KSR is a potent modulator of a signaling pathway essential to normal and oncogenic cell growth and development.
Assuntos
Substâncias de Crescimento/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno , Proteínas Quinases/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/metabolismo , Transdução de Sinais , Proteínas ras/metabolismo , Células 3T3 , Animais , Caenorhabditis elegans , Células Cultivadas , Drosophila , Ativação Enzimática , Insulina/farmacologia , MAP Quinase Quinase 1 , Camundongos , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
The intracellular mechanisms used by insulin and insulin-like growth factors to block programmed cell death are unknown. To identify receptor structures and signaling pathways essential for anti-apoptotic effects on cells, we have created a chimeric receptor (colony-stimulating factor-1 receptor/insulin receptor chimera (CSF1R/IR)) connecting the extracellular, ligand-binding domain of the colony-stimulating factor-1 (CSF-1) receptor to the transmembrane and cytoplasmic domains of the insulin receptor. Upon activation with CSF-1, the CSF1R/IR phosphorylates itself and intracellular substrates in a manner characteristic of normal insulin receptors. CSF-1 treatment protected cells expressing the CSF1R/IR from staurosporine-induced apoptosis. A chimeric receptor (CSF1R/IRDelta960) with a deletion of 12 amino acids from its juxtamembrane domain was constructed and expressed. CSF-1-treated cells expressing the CSF1R/IRDelta960 are unable to phosphorylate IRS-1 and Shc (Chaika, O. V., Chaika, N., Volle, D. J., Wilden, P. A. , Pirrucello, S. J., and Lewis, R. E. (1997) J. Biol. Chem. 272, 11968-11974). CSF-1 stimulated glucose uptake, mitogen-activated protein kinases, and IRS-1-associated phosphatidylinositol 3' kinase in cells expressing the CSF1R/IR but not in cells expressing the CSF1R/IRDelta960. Surprisingly, the CSF1R/IRDelta960 was as effective as the CSF1R/IR in mediating CSF-1 protection of cells from staurosporine-induced apoptosis. These observations indicate that the anti-apoptotic effects of the insulin receptor cytoplasmic domain can be mediated by signaling pathways distinct from those requiring IRS-1 and Shc.
Assuntos
Apoptose , Fator Estimulador de Colônias de Macrófagos/metabolismo , Receptor de Insulina/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Deleção de Sequência , Transdução de Sinais , Animais , Células CHO , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Membrana Celular/metabolismo , Cricetinae , Fragmentação do DNA , Ativação Enzimática , Glucose/metabolismo , Fator Estimulador de Colônias de Macrófagos/genética , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Receptor de Insulina/genética , Receptor de Fator Estimulador de Colônias de Macrófagos/genética , Receptor de Fator Estimulador de Colônias de Macrófagos/metabolismo , Proteínas Recombinantes de Fusão/genéticaRESUMO
A chimeric growth factor receptor (CSF1R/IR) was constructed by splicing cDNA sequences encoding the extracellular ligand binding domain of the human colony stimulating factor-1 (CSF-1) receptor to sequences encoding the transmembrane and cytoplasmic domains of the human insulin receptor. The addition of CSF-1 to cells transfected with the CSF1R/IR chimera cDNA stimulated the tyrosine phosphorylation of a protein that was immunoprecipitated by an antibody directed against the carboxyl terminus of the insulin receptor. Phosphopeptide maps of the 32P-labeled CSF1R/IR protein revealed the same pattern of phosphorylation observed in 32P-labeled insulin receptor beta subunits. CSF-1 stimulated the tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and Shc in cells expressing the CSF1R/IR chimera. Lipid accumulation and the expression of a differentiation-specific marker demonstrated that 3T3-L1 preadipocytes undergo CSF-1-dependent differentiation when transfected with the CSF1R/IR chimera cDNA but not when transfected with the expression vector alone. A 12-amino acid deletion within the juxtamembrane region of the CSF1R/IR (CSF1R/IRDelta960) blocked CSF-1-stimulated phosphorylation of IRS-1 and Shc but did not inhibit CSF-1-mediated differentiation of 3T3-L1 preadipocytes. These observations indicate that adipocyte differentiation can be initiated by intracellular pathways that do not require tyrosine phosphorylation of IRS-1 or Shc.
Assuntos
Adipócitos/citologia , Diferenciação Celular/efeitos dos fármacos , Fatores Estimuladores de Colônias/farmacologia , Receptor de Insulina/fisiologia , Receptores de Fator Estimulador de Colônias/fisiologia , Células 3T3 , Adipócitos/efeitos dos fármacos , Adipócitos/fisiologia , Animais , Células CHO , Cricetinae , Primers do DNA , Humanos , Cinética , Camundongos , Mapeamento de Peptídeos , Fosfopeptídeos/química , Fosfopeptídeos/isolamento & purificação , Fosforilação , Reação em Cadeia da Polimerase , Receptor de Insulina/biossíntese , Receptores de Fator Estimulador de Colônias/biossíntese , Proteínas Recombinantes de Fusão/metabolismo , TransfecçãoRESUMO
The enzyme immunoassay and neuraminidase activity inhibition test using polyclonal and monospecific antineuraminidase sera were employed to establish the similarities and differences in the antigenic structure of neuraminidase of influenza A viruses (H1N1), serovariant Hsw1N1, isolated from man in Alma-Ata (USSR), 1984, New Jersey (USA), 1976, and Pazardjik (BPR), 1982, as well as from swine and birds. Oligonucleotide mapping revealed significant structural differences in the genes coding for neuraminidase of Hsw1N1 viruses. The experimental results indicate a high degree of the enzyme variability in this group of viruses.
Assuntos
Antígenos Virais/análise , Epitopos/análise , Genes Virais , Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A/enzimologia , Neuraminidase/imunologia , Animais , Testes de Inibição da Hemaglutinação , Humanos , Vírus da Influenza A/genética , Vírus da Influenza A/imunologia , Neuraminidase/genética , Especificidade da EspécieRESUMO
Competitive radioimmunoassay was used to study the antigenic composition of hemagglutinin of Hsw1N1 viruses isolated from man in comparison with hemagglutinin Hsw1 of influenza virus of swine and ducks. The data of oligonucleotide analysis of the 4th RNA segment coding for hemagglutinin in these viruses are presented. It has been shown that in Alma-Ata, 1984-1985, influenza viruses Hsw1N1 were isolated with the antigenic structure of hemagglutinin and with the hemagglutinin gene identical with those of the classical influenza virus of swine A/Swine/Iowa/15/30 but differing from virus A/New Jersey/8/76.
Assuntos
Antígenos Virais/análise , Hemaglutininas Virais/análise , Vírus da Influenza A/imunologia , Adsorção , Animais , Antígenos Virais/genética , Ligação Competitiva , Patos , Genes Virais , Hemaglutininas Virais/genética , Humanos , Vírus da Influenza A/genética , Vírus da Influenza A/isolamento & purificação , Cazaquistão , Oligonucleotídeos/análise , Oligonucleotídeos/genética , RNA Viral/análise , RNA Viral/genética , Radioimunoensaio/métodos , SuínosRESUMO
Oligonucleotide mapping of individual genes was used for search of possible genetic recombinants between natural isolates of influenza H1N1 and H3N2 viruses isolated in the USSR in 1977-1979. No antigenic hybrids and recombinants with the antigenic structure H3N2 were found, however, it was shown that isolates of H1N1 viruses of 1979 (the A/USSR/61/79 strain) might represent genetic recombinants carrying genes P1 + P2 from H3N2 viruses, the M-gene of the USSR/61/79 virus being closest in its structure to the analogous gene of the earliest isolate of H3N2 viruses, namely A/Hong Kong/1/68. Possible selective advantages of virus recombinants having M-genes from viruses of a different serotype are discussed.
Assuntos
Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A Subtipo H3N2 , Vírus da Influenza A/genética , Recombinação Genética , Genes Virais , Vírus da Influenza A/classificação , Vírus da Influenza A/isolamento & purificação , Mapeamento de Nucleotídeos , Oligonucleotídeos/genética , RNA Viral/genética , SorotipagemRESUMO
Data are presented on structural variability of individual genes of selected variants of epidemic influenza viruses H1N1 (1977-1979) and H3N2 (1968-1979) in the course of antigenic drift obtained by oligonucleotide mapping. Six out of 8 genes of H1N1 viruses were found to be more variable than the corresponding genes of H3N2 viruses. Only HA and NS genes of H3N2 viruses underwent greater structural changes as compared with the analogous genes of H1N1 viruses. In viruses of both serotypes, most variable were the genes coding for hemagglutinin and matrix protein. Possible causes of greater structural variability of the matrix protein gene in the course of antigenic drift are discussed.
Assuntos
Variação Antigênica , Antígenos Virais/genética , Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A Subtipo H3N2 , Vírus da Influenza A/genética , Antígenos Virais/classificação , Surtos de Doenças , Genes Virais , Humanos , Vírus da Influenza A/classificação , Vírus da Influenza A/imunologia , Vírus da Influenza A/isolamento & purificação , Influenza Humana/microbiologia , Mapeamento de Nucleotídeos , Oligonucleotídeos/análise , RNA Viral/análise , RNA Viral/genética , SorotipagemRESUMO
The oligonucleotide mapping technique and RNA-RNA hybridization revealed structural differences between the HA- and M-protein genes of two variants of influenza A FPV, remantadine-sensitive and remantadine-resistant ones. A quantitative estimation of the structural divergence of the two influenza FPV pairs of genes was carried out. The possible functional role of the differences in the structure of the HA- and M-protein genes of the two FPV variants is discussed.
Assuntos
Adamantano/análogos & derivados , Genes Virais/efeitos dos fármacos , Variação Genética/efeitos dos fármacos , Hemaglutininas Virais/genética , Vírus da Influenza A/genética , Rimantadina/farmacologia , Proteínas da Matriz Viral/genética , Resistência Microbiana a Medicamentos/genética , Código Genético/efeitos dos fármacos , Vírus da Influenza A/efeitos dos fármacos , Hibridização de Ácido Nucleico , Oligopeptídeos/genética , Mapeamento de Peptídeos , RNA Viral/genéticaRESUMO
After 12 passages of a mouse-nonpathogenic influenza A/USSR/90/77 virus in mouse lungs a pathogenic virus was obtained causing death of the animals at 4-7 days after intranasal inoculation. The genetic and structural analysis of the initial and pathogenic viruses performed by oligonucleotide mapping of individual virus genes demonstrated that in the course of adaptation to mice structural changes had occurred at least in 5 out of 8 genes of virus with the exception of the genes coding for matrix and nonstructural proteins. The greatest differences were found in the genes coding for surface glycoproteins: hemagglutinin and neuraminidase. The experimental results indicate an important functional role of surface glycoproteins of influenza virus, particularly hemagglutinin, in the process of adaptation and formation of the pathogenic properties of virus.
Assuntos
Adaptação Biológica , Variação Genética , Vírus da Influenza A/genética , Animais , Mapeamento Cromossômico , Genes Virais , Glicoproteínas/genética , Humanos , Vírus da Influenza A/patogenicidade , Pulmão/microbiologia , Camundongos , Oligonucleotídeos/genética , Proteínas Virais/genética , VirulênciaRESUMO
A total preparation of cytoplasmic RNA was isolated from late stage-infected African green monkey kidney cells using phenol extraction procedure. The poly (A)-containing mRNA fraction was selected on oligo(dT)-cellulose columns. The resulting mRNA preparations were heterogenous in size and contained about 20--60% of SA7-derived sequences. SA7 late mRNA was efficiently translated in rabbit reticulocyte cell-free system giving rise to a number of polypeptide products that were related by antigenicity to authentic SA7 virion proteins. The main translation product having a molecular weight of 114 kilodaltons was identified as intact SA7 hexon protein.