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1.
Br J Cancer ; 104(1): 101-9, 2011 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-21119656

RESUMO

BACKGROUND: α-Tocopherol ether-linked acetic acid (α-TEA) is a promising agent for cancer prevention/therapy based on its antitumour actions in a variety of cancers. METHODS: Human breast cancer cells, MCF-7 and HCC-1954, were used to study the effect of α-TEA using Annexin V/PI staining, western blot analyses, and siRNA knockdown techniques. RESULTS: α-Tocopherol ether-linked acetic acid suppressed constitutively active basal levels of pAKT, pERK, pmTOR, and their downstream targets, as well as induced both cell types to undergo apoptosis. Phosphoinositide 3-kinase (PI3K) inhibitor wortmannin suppressed pAKT, pERK, pmTOR, and their downstream targets, indicating PI3K to be a common upstream mediator. In addition, α-TEA induced increased levels of pIRS-1 (Ser-307), a phosphorylation site correlated with insulin receptor substrate-1 (IRS-1) inactivation, and decreased levels of total IRS-1. Small interfering RNA (siRNA) knockdown of JNK blocked the impact of α-TEA on pIRS-1 and total IRS-1 and impeded its ability to downregulate the phosphorylated status of AKT, ERK, and mTOR. Combinations of α-TEA+MEK or mTOR inhibitor acted cooperatively to induce apoptosis and reduce basal levels of pERK and pmTOR. Importantly, inhibition of MEK and mTOR resulted in increased levels of pAKT and IRS-1, and α-TEA blocked them. CONCLUSIONS: Downregulation of IRS-1/PI3K pathways via JNK are critical for α-TEA and α-TEA+MEK or mTOR inhibitor-induced apoptosis in human MCF-7 and HCC-1954 breast cancer cells.


Assuntos
1-Fosfatidilinositol 4-Quinase/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Neoplasias da Mama/patologia , Proteínas Substratos do Receptor de Insulina/antagonistas & inibidores , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , Serina-Treonina Quinases TOR/antagonistas & inibidores , alfa-Tocoferol/farmacologia , 1-Fosfatidilinositol 4-Quinase/metabolismo , Androstadienos/farmacologia , Antracenos/farmacologia , Antioxidantes/farmacologia , Western Blotting , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Butadienos/farmacologia , Sinergismo Farmacológico , Quimioterapia Combinada , Inibidores Enzimáticos/farmacologia , Feminino , Humanos , Imunossupressores/farmacologia , Proteínas Substratos do Receptor de Insulina/metabolismo , MAP Quinase Quinase 4/antagonistas & inibidores , MAP Quinase Quinase 4/genética , MAP Quinase Quinase 4/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Nitrilas/farmacologia , Fosforilação/efeitos dos fármacos , RNA Interferente Pequeno/genética , Transdução de Sinais/efeitos dos fármacos , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/metabolismo , Células Tumorais Cultivadas , Wortmanina
2.
Anim Reprod Sci ; 221: 106581, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32891911

RESUMO

Freezing cooled-transported semen allows veterinarians and breeders to collect and process the semen of stallions on farm, and then ship the semen to a semen freezing center. There, however, is a lack of standardization of shipping and freezing protocols. The objectives were to optimize and simplify protocols to freeze cooled-shipped semen. In Experiment 1, cooled-transported semen was centrifuged at room temperature or 5 °C before freezing. Sperm variables (motility, membrane integrity, acrosome integrity, membrane fluidity) were evaluated before and after freezing. Centrifugation temperature had no effect on post-thaw semen quality. In Experiment 2, cooled-transported semen was centrifuged at room temperature and cryopreserved in three semen freezing extenders. With use of the improved modified French formula, there was less post-thaw total and progressive motility compared with use of Botucrio or the improved lactose-EDTA formula (P<0.0001). Semen cryopreserved in the improved modified French formula also had a lesser percentage of sperm with intact membranes compared with lactose-EDTA, and a greater percentage of sperm with capacitation-like changes compared with Botucrio (P<0.0001). In Experiment 3, semen diluted in each extender was frozen conventionally or placed directly in a -80 °C ultra-freezer. Freezing in the ultra-freezer resulted in a lesser post-thaw sperm motility, but not membrane and acrosome integrity and capacitation-like changes. In conclusion, centrifugation and addition of freezing extender to cooled transported semen can be performed at room temperature or 5 °C. The Botucrio and lactose-EDTA formula are recommended for conventional cryopreservation of cooled-transported stallion semen as compared with the modified French formula.


Assuntos
Criopreservação/veterinária , Cavalos/fisiologia , Preservação do Sêmen/veterinária , Manejo de Espécimes/veterinária , Animais , Crioprotetores/farmacologia , Congelamento , Masculino , Sêmen/efeitos dos fármacos , Manejo de Espécimes/métodos
3.
Cancer Res ; 50(15): 4764-70, 1990 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-2142445

RESUMO

Infection of animals with retroviruses frequently leads to immunosuppressed states. The immune status of chickens injected with the replication-defective avian erythroblastosis virus (AEV), with its naturally occurring subgroup B helper virus (avian erythroblastosis-associated virus; AEAV), was evaluated daily and compared to the immune status of age-matched uninfected control chickens. Spleen cells from AEV-infected chickens gave depressed responses to concanavalin A, phytohemagglutinin, and pokeweed mitogen beginning 3 days after injection of the virus and continuing until death. Spleen cells from AEV-infected chickens suppressed the T-cell mitogen-induced blastogenic responses of spleen cells from uninfected chickens. The ability of spleen cells from infected chickens to suppress mitogen-induced blastogenic responses of spleen cells from normal chickens in coculture was transient beginning 4 days following viral inoculation, reaching peak levels of suppression on day 7 and disappearing by day 12. Cytolysis of splenic cells from AEV-infected chickens with polyclonal anti-T-cell-serum removed the suppressor activity. Addition of conditioned medium rich in T-cell growth factor resulted in a partial restoration of the blastogenic responsiveness of splenic cells from 6-day post-AEV-infected chickens. Addition of exogenous T-cell growth factor had no effect on the suppressed blastogenic responsiveness of spleen cells from 12-day post-AEV-infected chickens, and it had no effect on coculture suppression. In addition to suppressed T-cell responses to polyclonal mitogen-induced proliferation in vitro and transiently expressed T-suppressor cells, thymic atrophy and structural disruption was observed in AEV-infected chickens.


Assuntos
Leucose Aviária/imunologia , Ativação Linfocitária , Linfócitos/imunologia , Alpharetrovirus , Animais , Leucose Aviária/patologia , Bolsa de Fabricius/patologia , Adesão Celular , Células Cultivadas , Galinhas , Interleucina-2/análise , Interleucina-2/biossíntese , Depleção Linfocítica , Valores de Referência , Baço/imunologia , Linfócitos T Reguladores/imunologia , Timo/patologia
4.
Cancer Res ; 42(11): 4532-9, 1982 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-6751524

RESUMO

Hybridoma cell lines secreting antibodies to chicken fetal antigens (CFAs) were generated by the fusion of mouse P3X63Ag8 myeloma cells with spleen cells from a mouse immunized with intact SC chicken strain one-day-hatched red blood cells. Immunodepletion studies show monoclonal anti-CFA to be detecting a subset of the Mr 50,000 CFA molecules recognized by polyclonal anti-CFA. Monoclonal anti-CFA is erythroid specific against in vivo-derived hematopoietic-lymphoid cells. Exceptions to the erythroid specificity of monoclonal anti-CFA include failure to react with avian erythroblastosis virus-transformed erythroid cells both before and after butyric acid-induced differentiation and reactions with reticuloendotheliosis virus-transformed immature lymphoid cells and chicken embryo cells. Immunofluorescence and 125I binding analyses utilizing monoclonal anti-CFA show reticuloendotheliosis virus cells to possess high levels of CFA even though the CFA determinant does not appear to be a 125I-labeled immunoprecipitable Mr 50,000 molecule. The unique property of monoclonal anti-CFA that permits it to distinguish among surface membrane antigens of normal and neoplastic cells of the same lineage makes it an important tool for future investigations of normal and abnormal cell differentiation.


Assuntos
Anticorpos Monoclonais , Antígenos de Superfície/análise , Medula Óssea/imunologia , Eritrócitos/imunologia , Células-Tronco Hematopoéticas/imunologia , Antígenos de Histocompatibilidade Classe II/análise , Linfócitos/imunologia , Animais , Complexo Antígeno-Anticorpo , Diferenciação Celular , Galinhas , Envelhecimento Eritrocítico , Imunofluorescência , Hibridomas/imunologia , Técnicas Imunoenzimáticas , Camundongos , Plasmocitoma/imunologia , Proteína Estafilocócica A
5.
Cancer Res ; 42(11): 4625-30, 1982 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-7127300

RESUMO

Avian erythroblastosis virus strain R (AEV)-transformed, cloned erythroleukemia cells from three different ages of SC strain chickens were analyzed before and after differentiation induced by 1.0 mM butyric acid for expression of chicken fetal antigens (CFAs) and chicken adult antigens (CAAs) and for hemoglobin expression. Immunofluorescent analyses show the loss of individual CFA determinants from erythroleukemia cells with induced differentiation, although there appeared to be no correlation between CFA loss and onset of hemoglobin production. Erythroleukemia cells were examined by cell surface labeling followed by immunoprecipitation with antisera specific to CFAs and CAAs. Erythroleukemia cells expressed CFAs and CAAs on their membranes that are not reported to be expressed by the target cell of AEV. The expression of CAAs and the enhanced expression of CFAs by erythroleukemia cells may be due to limited cellular differentiation, alterations in regulatory controls of genes coding for CFAs and CAAs, or increased levels of production of previously undetected CFAs and CAAs following AEV transformation. Control and induced erythroleukemia cells expressed CFAs and CAAs that differed both quantitatively and qualitatively from normal erythroid cells. Molecular weight variations of CFAs and CAAs observed in the erythroleukemia cells may represent glycolyzation differences between AEV-transformed cells and normal erythroid cells.


Assuntos
Antígenos de Superfície/análise , Leucose Aviária/imunologia , Alpharetrovirus/imunologia , Animais , Leucose Aviária/microbiologia , Medula Óssea/fisiologia , Diferenciação Celular , Transformação Celular Neoplásica , Galinhas , Células Clonais
6.
Cancer Res ; 44(8): 3576-83, 1984 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-6378372

RESUMO

Hematopoietic-lymphoid membrane antigens that are related to cell differentiation and development, referred to as chicken fetal antigen (CFA) and chicken adult antigen (CAA) were immunochemically characterized; Mr 220,000; Mr 170,000; Mr 130,000; Mr 99,000; Mr 88,000; Mr 50,000; and Mr 24,000 CFA molecules are detected on embryonic RBC, and Mr 210,000; Mr 130,000; Mr 102,000; Mr 56,000; Mr 48,000; and Mr 43,000 CAA molecules are detected on adult RBC. Limited peptide mapping analyses showed all of the CFA and CAA molecules to be distinct entities. Both the Mr 50,000 CFA and the Mr 43,000 CAA molecules exhibited multiple isomorphic variants when analyzed by 2-dimensional electrophoresis. Analyses involving neuraminidase treatments and limited peptide mapping showed the Mr 50,000 CFA isomorphic variants to be chemically identical with the isoelectric point variations being due to sialic acid differences. In addition to multiple isomorphic variants, the molecular weight and charge differences of which were diminished by neuraminidase treatments, the Mr 43,000 CAA molecules exhibited a doublet pattern suggesting that the polyclonal antisera may be detecting chicken major histocompatibility complex products. Analyses of the Mr 50,000 CFA molecules immunoprecipitated with monoclonal antibody 190-4 confirmed that the monoclonal antibody recognizes a serological subset of the Mr 50,000 CFA molecules but showed that it did not recognize a unique molecularly detectable subset among the 18 isomorphic variants discernable by 2-dimensional electrophoretic analyses. Cocapping analyses with splenic lymphocytes showed CFA and CAA to occur as distinct membrane entities on lymphocytes.


Assuntos
Antígenos de Superfície/análise , Envelhecimento Eritrocítico , Membrana Eritrocítica/imunologia , Animais , Anticorpos , Anticorpos Monoclonais , Complexo Antígeno-Anticorpo , Diferenciação Celular , Galinhas , Eletroforese em Gel de Poliacrilamida , Técnicas Imunoenzimáticas , Complexo Principal de Histocompatibilidade , Peso Molecular , Neuraminidase , Fragmentos de Peptídeos/análise
7.
Cancer Res ; 61(17): 6569-76, 2001 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-11522656

RESUMO

RRR-alpha-tocopherol succinate (vitamin E succinate, VES) is a potent, selective apoptotic agent for cancer cells but not normal cells. VES has been shown to inhibit the growth of a wide variety of tumor cells in cell culture and animal models. Studies addressing mechanisms of action of VES-induced apoptosis have identified transforming growth factor-beta, Fas/CD95-APO-1, and mitogen-activated protein kinase (MAPK) signaling pathway involvement. Here we show that MAPKs, the extracellular signal-regulated kinases (ERK), and the stress-activated protein kinases, c-Jun NH2-terminal kinases (JNK), but not p38, are critical mediators in VES-induced apoptosis of human breast cancer MDA-MB-435 cells. VES activates ERK1/2 and JNK both in level and duration of kinase activity. Expression of dominant negative mutants of ERK1, MAPK/ERK activator-1, or JNK1 but not p38 blocked phosphorylation of the substrate glutathione S-transferase-c-Jun and inhibited VES-induced apoptosis. Increased phosphorylation and transactivation activity of nuclear transcription factors c-Jun, ATF-2, and Elk-1 are observed after VES treatments; however, only c-Jun and ATF-2 appear to be involved in VES-induced apoptosis based on antisense blockage experiments. Collectively, these results imply a critical role for ERK1 and JNK1 but not p38 in VES-induced apoptosis of human MDA-MB-435 breast cancer cells.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias da Mama/enzimologia , Proteínas de Ligação a DNA , Proteínas Quinases JNK Ativadas por Mitógeno , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Vitamina E/análogos & derivados , Vitamina E/farmacologia , Fator 2 Ativador da Transcrição , Apoptose/fisiologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Ativação Enzimática , Humanos , MAP Quinase Quinase 4 , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno , Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/fisiologia , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/farmacologia , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Tocoferóis , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Ativação Transcricional/efeitos dos fármacos , Células Tumorais Cultivadas , Proteínas Elk-1 do Domínio ets , Proteínas Quinases p38 Ativadas por Mitógeno
8.
Cancer Res ; 59(4): 953-61, 1999 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-10029090

RESUMO

Fas (CD95/APO-1) is an important mediator of apoptosis. We show that Fas-resistant MCF-7, MDA-MB-231, and MDA-MB-435 human breast cancer cells become responsive to anti-Fas (CD95) agonistic antibody-triggered apoptosis after pretreatment or cotreatment with vitamin E succinate (VES; RRR-alpha-tocopheryl succinate). In contrast, no enhancement of anti-Fas agonistic antibody-triggered apoptosis was observed following VES pretreatment or cotreatment with Fas-sensitive primary cultures of human mammary epithelial cells, immortalized MCF-10A cells, or T47D human breast cancer cells. Although VES is itself a potent apoptotic triggering agent, the 6-h pretreatment procedure for Fas sensitization did not initiate VES-mediated apoptosis. The combination of VES plus anti-Fas in pretreatment protocols was synergistic, inducing 2.8-, 3.0-, and 6.3-fold enhanced apoptosis in Fas-resistant MCF-7, MDA-MB-231, and MDA-MB-435 cells, respectively. Likewise, cotreatment of Fas-resistant MCF-7, MDA-MB-231, and MDA-MB-435 cells with VES plus anti-Fas enhanced apoptosis 1.9-, 2.0-, and 2.6-fold, respectively. Functional knockout of Fas-mediated signaling with either Fas-neutralizing antibody (MCF-7-, MDA-MB-231-, and MDA-MB-435-treated cells) or Fas antisense oligomers (MDA-MB-435-treated cells only), reduced VES-triggered apoptosis by approximately 50%. Analyses of whole cell extracts from Fas-sensitive cells revealed high constitutive expression of Mr 43,000 Fas, whereas Fas-resistant cells expressed low levels that were confined to the cytosolic fraction. VES treatment of the Fas-resistant cells caused a depletion of cytosolic Mr 43,000 Fas with a concomitant increase in Mr 43,000 membrane Fas. These data show that VES can convert Fas-resistant human breast cancer cells to a Fas-sensitive phenotype, perhaps by translocation of cytosolic Mr 43,000 Fas to the membrane and show that VES-mediated apoptosis involves Mr 43,000 Fas signaling.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias da Mama/patologia , Proteínas Quinases JNK Ativadas por Mitógeno , Quinases de Proteína Quinase Ativadas por Mitógeno , Vitamina E/análogos & derivados , Receptor fas/fisiologia , Feminino , Humanos , Interferon gama/farmacologia , MAP Quinase Quinase 4 , Peso Molecular , Proteínas Quinases/fisiologia , Tocoferóis , Fator de Crescimento Transformador beta/fisiologia , Células Tumorais Cultivadas , Vitamina E/farmacologia , Receptor fas/análise
9.
Theriogenology ; 85(7): 1219-24, 2016 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-26764152

RESUMO

Urospermia is known to affect semen quality in many mammals, including stallions. Determinations of semen pH and creatinine and urea concentrations have been used to diagnose urine contamination in raw stallion semen. Unfortunately, practitioners suspecting urine contamination in cooled-shipped samples have no proven means to confirm the presence of urine. Therefore, the objectives of this study were (1) to assess the effects of urine contamination on sperm motility of extended fresh and cooled-stored stallion semen, (2) to evaluate the usefulness of semen color, odor, pH, and creatinine and urea concentrations for urospermia diagnosis, and (3) to evaluate the accuracy of a commercial blood urea nitrogen test strip in diagnosing urine contamination in extended-cooled stallion semen. Thirty-seven ejaculates were obtained from 11 stallions with no history of urospermia before division into 5 mL aliquots, and contamination with stallion urine. Each resulting sample was assessed for sperm motility, color, odor, pH, creatinine, and urea nitrogen concentration using both a semiquantitative test strip (Azostix), and a quantitative automated analyzer before and after cooling for 24 hour. Sperm motility parameters, pH, and creatinine and urea concentrations were analyzed using mixed models. Urine contamination decreased total and progressive motility in all samples before and after cooling (P < 0.05). Mean control total motility was 80% at 0 hour and 67% at 24 hours, whereas urine-contaminated samples ranged from 30% to 71% at 0 hour and 27% to 61% at 24 hours. Control mean urea (29 mg/dL) and creatinine (0.6 mg/dL) concentrations were significantly different (P < 0.05) from all urine-contaminated samples (158 mg/dL and 11.6 mg/dL, respectively) at 0 hour. Similarly, control mean urea (8 mg/dL) and creatinine (0.9 mg/dL) concentrations were significantly different than all urine-contaminated samples at 24 hours. Odor assessment presented moderate sensitivity (65%) and high specificity (100%), while color assessment presented low sensitivity (47%) and moderate specificity (79%) for urine in extended semen. Azostix strips were highly sensitive (95%) and specific (97%). Assessment of color, odor, and pH are not reliable methods to diagnose urine in experimentally contaminated cooled-stored stallion semen. Sperm motility parameters (in raw and cooled semen) are significantly reduced by the presence of urine in a concentration dependent. The results of the present study indicated that determination of urea and creatinine concentrations can be used to diagnose urospermia and that Azostix can be used as a point care method for diagnosing urine contamination in extended cooled stallion semen.


Assuntos
Cavalos/urina , Preservação do Sêmen/veterinária , Manejo de Espécimes/veterinária , Urina , Animais , Cavalos/fisiologia , Masculino
10.
Vet Rec ; 178(10): 241, 2016 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-26908160

RESUMO

This study compares two methods for seminal plasma removal by evaluating sperm recovery rates, and motility and morphology of cooled-stored semen. Ejaculates were divided into three groups: control, filtration and cushioned centrifugation. Semen was extended to 25 million sperm/ml using a skim-milk-based extender and stored at 5°C for all groups. Sperm motility (total motility (%TM) and progressive motility (%PM)) was determined at 0, 24, 48 and 72 hours by a computer-assisted sperm analyser. Sperm morphology was assessed using differential interference microscopy. Overall, %TM of the centrifugation group was significantly higher than the filter group, but not significantly different than the control. No significant difference in %TM or %PM was detected for the control group and filter. Cushioned centrifugation was a superior method to obtain progressively motile sperm compared with control (P=0.03) and filter groups (P<0.001). No significant difference was found for the per cent of normal sperm cells and detached heads between the groups. This study demonstrated that cushioned centrifugation was a superior method to remove seminal plasma while preserving %TM and enhancing %PM for stallions under cooled storage over three days. However, as the differences appear to be negligible, the SpermFilter may represent an alternative for farms lacking a centrifuge.


Assuntos
Centrifugação/veterinária , Criopreservação/veterinária , Filtração/veterinária , Preservação do Sêmen/veterinária , Espermatozoides/citologia , Espermatozoides/fisiologia , Animais , Centrifugação/métodos , Filtração/métodos , Cavalos , Longevidade , Masculino , Preservação do Sêmen/métodos , Motilidade dos Espermatozoides/fisiologia
11.
Oncogene ; 15(2): 223-30, 1997 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-9244357

RESUMO

Previous studies have shown that treatment of avian reticuloendotheliosis virus-transformed RECC-UTC4-1 (C4-1) lymphoblastoid cells with 10 microg/ml (18.8 microM) of RRR-alpha-tocopheryl succinate (vitamin E succinate, VES) for 3 days induced approximately 50% of the cells to undergo apoptosis. Elevated and prolonged expression of c-jun mRNA and protein was temporally correlated with VES-induced cell death. Data presented in this paper show that the elevated and prolonged expression of c-jun message and protein are not accounted for by enhanced stability, and show the involvement of c-Jun in VES-induced apoptosis in this lymphoblastoid cell type. C4-1 cells infected with a virus carrying a dominant, negatively acting mutant form of c-Jun, supjun-1, exhibited: (i) 71% reduction in VES-induced apoptosis, (ii) a 2.0-2.5-fold decrease in wildtype, endogenous c-Jun expression, and (iii) a 2.4-2.6-fold reduction in AP-1 binding activity. Additionally, cells co-treated with VES plus RRR-alpha-tocopherol, exhibited a 70% reduction in apoptosis, a marked reduction in c-Jun expression and a 1.6-fold reduction in AP-1 binding activity. These studies suggest that c-Jun plays a crucial role in VES-induced apoptosis in C4-1 cells, and add to our understanding of mechanisms of action involved in VES-mediated tumor cell growth inhibition.


Assuntos
Apoptose/efeitos dos fármacos , Transformação Celular Neoplásica , Proteínas Proto-Oncogênicas c-jun/fisiologia , Vírus da Reticuloendoteliose/genética , Vitamina E/análogos & derivados , Animais , Linhagem Celular , Galinhas , Linfócitos/efeitos dos fármacos , RNA Mensageiro/análise , Tocoferóis , Vitamina E/farmacologia
12.
Plant Physiol ; 104(2): 613-616, 1994 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12232111

RESUMO

When two different forms of phosphoenolpyruvate carboxylase (PEPC) from maize (Zea mays L.) leaves are present in an assay it is possible to estimate the ratio of Vmax to Km (V/K) for the two forms separately. This measure of the binding of the substrate by the enzyme permits evaluation of the effects of various treatments on the relative substrate-binding velocity of the enzyme. PEPC diluted 1/20 is present in a mixture of a tetrameric form with a high affinity for phosphoenolpyruvate and a dimeric form with a low affinity (M.-X. Wu, C.R. Meyer, K.O. Willeford, R.T. Wedding [1990] Arch Biochem Biophys 281: 324-329). Malate at 5 mM reduced (V/K)1,[mdash]the V/K of the probable tetrameric form[mdash]almost to zero, but reduced (V/K)2[mdash]the V/K of the probable dimer[mdash]by only about 80%. Glucose-6-phosphate (Glc-6-P) at 5 mM increased (V/K)1 to 155% of the control but had no effect on (V/K)2. Glycerol (20%) alone increased both V/Ks, and its effects are additive to the Glc-6-P effects, implying different mechanisms for activation by Glc-6-P and glycerol.

13.
Dev Comp Immunol ; 14(4): 425-37, 1990.
Artigo em Inglês | MEDLINE | ID: mdl-2086272

RESUMO

Monoclonal antibody R7-3 recognized an erythroid specific cell surface molecule with a m.w. of approximately 98 kilodaltons (Kd) under nonreducing conditions and molecules of 40 and 44 Kd under reducing conditions on both embryonic- and adult-derived peripheral RBC. Immunochemical characterization, including limited peptide map analyses of these molecules, provided evidence that mAb R7-3 was recognizing the MHC coded B-G heterodimer. This is the first report of a monoclonal antibody that recognizes the B-G heterodimer. Affinity binding studies suggested that mAb R7-3 preferentially recognized the 44 Kd molecule. Immune depletion analyses demonstrated the presence of a single population of B-G heterodimers. Endoglycosidase-F and neuraminidase digestions suggested that the 44 and 40 Kd molecules contained very little, if any, carbohydrate. B-G heterodimer expression was examined on primitive and definitive RBC during embryonic development. B-G heterodimer expression was not detected on RBC of other avians.


Assuntos
Galinhas/imunologia , Antígenos de Histocompatibilidade/isolamento & purificação , Animais , Anticorpos Monoclonais , Embrião de Galinha/imunologia , Galinhas/sangue , Eritrócitos/imunologia , Glicosilação , Antígenos de Histocompatibilidade/química , Antígenos de Histocompatibilidade/metabolismo , Mapeamento de Peptídeos
14.
Dev Comp Immunol ; 13(3): 231-52, 1989.
Artigo em Inglês | MEDLINE | ID: mdl-2792501

RESUMO

Antiserum prepared against sucrose gradient purified reticuloendotheliosis virus (REV) recognized the chicken transferrin receptor. Molecules immunoprecipitated from red blood cells (RBC) obtained from embryonic chickens with either the anti-REV reagent or a chicken transferrin immunomatrix were demonstrated to be identical by co-migration in both reducing and nonreducing SDS-polyacrylamide gels and in two-dimensional isoelectric focusing analyses, reciprocal immunodepletion analyses and by peptide mapping. The chicken transferrin receptor was shown to be a 190,000 dalton cell surface membrane molecule consisting of two similar disulfide-bonded subunits of approximately 95,000 daltons. The chicken transferrin receptor was expressed on erythroid cell surface membranes as 95,000 dalton monomers as well as 190,000 dalton dimers. The chicken transferrin receptor was expressed on all differentiation/maturation stages, including mature RBC, of both the primitive and definitive type I erythroid cell series. In adult chickens, the transferrin receptor was expressed by immature erythroid cells in the bone marrow, but not by mature circulating RBC. REV-transformed immature lymphoid cells and avian erythroblastosis virus (AEV)-transformed erythroid cells expressed dimers composed of 95,000 and 110,000 dalton subunits. Comparisons among V8 protease derived peptides from 95,000 dalton transferrin receptors obtained from RBC and REV-transformed lymphoid cells revealed a high degree of homology; however, the 95,000 dalton molecules isolated from REV-transformed lymphoid cells exhibited a 56,000 dalton peptide that was unique. Cloned AEV-transformed erythroleukemia cells induced to differentiate by supplementation of the media with 1 mM butyric acid expressed elevated transferrin receptor levels. Both serological and peptide mapping studies demonstrated the human transferrin receptor on K562 cells and the chicken transferrin receptor to be distinct. However, chicken transferrin was shown to be capable of reacting with the human transferrin receptors on K562 cells.


Assuntos
Galinhas/metabolismo , Receptores da Transferrina/metabolismo , Transferrina/metabolismo , Animais , Transformação Celular Viral , Embrião de Galinha , Eritrócitos/metabolismo , Eritropoese , Humanos , Peso Molecular , Receptores da Transferrina/isolamento & purificação , Retroviridae
15.
J Appl Physiol (1985) ; 68(4): 1399-404, 1990 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-2347782

RESUMO

The purpose of this study was to investigate metabolic changes in equine muscle from birth to 1 yr of age. Duplicate biopsies from the middle portion of the gluteus medius were obtained from a depth of 2 cm beneath the superficial fascia at 1 day, 7 days, 1 mo, 3 mo, 6 mo, and 1 yr of age in 11 quarter horses and at 1 day, 3 mo, 6 mo, and 1 yr of age in 5 Standardbreds. Muscle enzyme activities determined were citrate synthase, 3-hydroxyacyl-CoA dehydrogenase, phosphorylase, and lactate dehydrogenase. Percent fast-twitch, fast-twitch high oxidative, and slow-twitch oxidative fiber types were determined using succinate dehydrogenase and myosin adenosinetriphosphatase (pH 9.4) histochemical stains. Histochemically determined muscle fiber-type percents did not change dramatically with increasing age. However, lactate dehydrogenase activity increased threefold in quarter horses and twofold in Standardbreds, and phosphorylase activity increased sixfold in quarter horses and sevenfold in Standardbreds from 1 day to 6 mo of age. Citrate synthase and 3-hydroxyacyl-CoA dehydrogenase activities decreased during the first 3 mo of age in quarter horses.


Assuntos
Envelhecimento/metabolismo , Cavalos/fisiologia , Músculos/enzimologia , 3-Hidroxiacil-CoA Desidrogenases/metabolismo , Animais , Biópsia , Citrato (si)-Sintase/metabolismo , L-Lactato Desidrogenase/metabolismo , Desenvolvimento Muscular , Músculos/anatomia & histologia , Fosforilases/metabolismo
16.
J Am Diet Assoc ; 87(4): 463-8, 1987 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3559005

RESUMO

The purpose of this study was to investigate the impact of eating away from home (at commercial establishments) on nutrient adequacy by examining frequency of eating away from home, the nutritional value of foods eaten both away and at home, and the nutritional adequacy of the daily diets of individuals. Data from 3,500 individuals, 15 years of age or older, interviewed in the fall quarter of the 1977-78 Nationwide Food Consumption Survey were used. Results indicate that although the nutrient density of food eaten away from home was lower than that of food eaten at home, the persons studied did not eat out frequently enough to influence the adequacy of their diets significantly. The low nutrient density of food eaten away from home does suggest, however, that individuals could be putting themselves at risk of some nutrient inadequacies (particularly of calcium and vitamins A, B-6, and C) or of caloric excess if they substantially increase their frequency of eating away from home. Teenagers and senior citizens seem most vulnerable to potential nutritional inadequacies.


Assuntos
Inquéritos sobre Dietas , Inquéritos Nutricionais , Necessidades Nutricionais , Adolescente , Adulto , Fatores Etários , Idoso , Ingestão de Energia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Minerais , Valor Nutritivo , Restaurantes , Fatores Sexuais , Vitaminas
17.
Soc Sci Med ; 51(12): 1805-16, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11128268

RESUMO

An ideology of uncertainty reduction pervades scholarly and popular discourse on breast self-exams (BSE). Women are encouraged to understand BSE as an activity that reduces uncertainty about their health. Moreover, uncertainties about the procedure itself are conceived as barriers to BSE. In turn, reducing these uncertainties is seen as the key to promoting BSE. We argue that the ideology of uncertainty reduction is both descriptively and prescriptively inadequate and potentially a threat to women's health. We further contend the ideology should be replaced by a framework that illuminates processes of coping with uncertainty. Several major characteristics of such a framework, as well as implications for medical practice, are discussed and illustrated within the context of BSE.


Assuntos
Adaptação Psicológica , Autoexame de Mama , Comportamentos Relacionados com a Saúde , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/prevenção & controle , Feminino , Pesquisa sobre Serviços de Saúde , Humanos , Meios de Comunicação de Massa , Modelos Teóricos , Saúde da Mulher
18.
Int J Psychophysiol ; 37(2): 207-17, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10832007

RESUMO

Due to the importance of baseline and recovery levels in the computation of reactivity, two studies were conducted to determine an alternative method to traditional rest for achieving baseline and recovery levels of cardiovascular measurements. Watching a relaxing, aquatic video was compared with a traditional resting baseline to determine the better method for achieving low baseline levels. In addition, watching the video was compared with traditional rest during 5-min post-task recovery periods. Systolic (SBP) and diastolic blood pressure (DBP) decreased more during the baseline period when subjects viewed the video than when subjects rested quietly. Similarly, subjects displayed greater recovery following the mental tasks when they watched a video than when they merely sat quietly. It is recommended that researchers standardize baseline procedures by showing a relaxing video before administering tasks for the assessment of cardiovascular reactivity.


Assuntos
Hemodinâmica/fisiologia , Terapia de Relaxamento , Adulto , Pressão Sanguínea/fisiologia , Débito Cardíaco/fisiologia , Feminino , Humanos , Masculino , Desempenho Psicomotor/fisiologia , Gravação de Videoteipe
19.
Int J Psychophysiol ; 29(3): 291-301, 1998 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9666382

RESUMO

T-Wave Amplitude (TWA) has been suggested as an indicator of sympathetic influence on myocardial performance, but critics have argued that TWA is confounded by parasympathetic influence or that it is a non-specific feature of tachycardia. To help clarify the issue, we examined TWA as a function of parasympathetic activity, using cardiac vagal control as measured by high frequency components of heart period variability (respiratory sinus arrhythmia) and of interbeat intervals (IBI), across several stressful tasks. Sixteen male subjects were exposed to Valsalva, Serial Subtraction and Cold-Pressor tasks. After controlling for between-person variance, it was found that RSA did not contribute to TWA and that IBI contributed dependably to TWA only during the Valsalva maneuver, when heart rate was driven very high. In light of these results, we recommend that TWA continue to be considered a candidate indicator of sympathetic influence on myocardial performance, although caution should be used if heart rate is dramatically elevated.


Assuntos
Arritmia Sinusal/fisiopatologia , Eletrocardiografia , Frequência Cardíaca/fisiologia , Adulto , Temperatura Baixa , Humanos , Masculino , Pressão , Manobra de Valsalva/fisiologia
20.
Int J Psychophysiol ; 40(2): 93-107, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11165348

RESUMO

The present study delineates a method for the quantification of six hemodynamic reactivity patterns, in response to a laboratory stressor, and examines the psychophysiological correlates of individual differences in these patterns. One hundred and ninety-four young adult men and women participated in rest periods and two laboratory stressors, mental arithmetic and an anger recall interview. Measures were taken of blood pressure, heart rate, and cardiac output, from which total peripheral resistance was derived, as well as state reports of feelings during the tasks. Six hemodynamic reactor patterns were identified: Non-reactors, Mild Myocardials, Mild Vasculars, Myocardials, Vasculars, and Dual Reactors, each associated with a unique profile of cardiac output and total peripheral resistance change. Myocardial reactors to the interview had the highest resting levels of blood pressure and total peripheral resistance. Dual reactors had the largest increases in diastolic reactivity; Dual and Myocardial reactors had the largest increases in systolic reactivity. The extreme reactor groups (Dual, Myocardial, Vascular) all reported greater task invigoration than the Non-reactors, who reported greater efforts to relax. Reactor groups were similar on anger-related trait affect. Based on both resting blood pressure and magnitude of task-induced reactivity, Myocardial and Dual reactors may be at the greatest risk for subsequent hypertension.


Assuntos
Hemodinâmica/fisiologia , Estresse Psicológico/fisiopatologia , Estresse Psicológico/psicologia , Adolescente , Adulto , Ira/fisiologia , Pressão Sanguínea/fisiologia , Débito Cardíaco/fisiologia , Emoções/fisiologia , Feminino , Hostilidade , Humanos , Individualidade , Masculino , Processos Mentais/fisiologia , Rememoração Mental/fisiologia , Personalidade , Psicofisiologia , Inquéritos e Questionários , Resistência Vascular/fisiologia
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