RESUMO
Nitrogen and sulfur emissions from oil sands operations in northern Alberta, Canada have resulted in increasing deposition of N and S to the region's ecosystems. To assess whether a changing N and S deposition regime affects bog porewater chemistry, we sampled bog porewater at sites at different distances from the oil sands industrial center from 2009 to 2012 (10-cm intervals to a depth of 1 m) and from 2009 to 2019 (top of the bog water table only). We hypothesized that: (1) as atmospheric N and S deposition increases with increasing proximity to the oil sands industrial center, surface porewater concentrations of NH4+, NO3-, DON, and SO42- would increase and (2) with increasing N and S deposition, elevated porewater concentrations of NH4+, NO3-, DON, and SO42- would be manifested increasingly deeper into the peat profile. We found weak evidence that oil sands N and S emissions affect bog porewater NH4+-N, NO3--N, or DON concentrations. We found mixed evidence that increasing SO42- deposition results in increasing porewater SO42- concentrations. Current SO42- deposition, especially at bogs closest to the oil sands industrial center, likely exceeds the ability of the Sphagnum moss layer to retain S through net primary production, such that atmospherically deposited SO42- infiltrates downward into the peat column. Increasing porewater SO42- availability may stimulate dissimilatory sulfate reduction and/or inhibit CH4 production, potentially affecting carbon cycling and gaseous fluxes in these bogs.
Assuntos
Campos de Petróleo e Gás , Áreas Alagadas , Alberta , Ecossistema , Monitoramento Ambiental , ÁguaRESUMO
Increasing gaseous emissions of nitrogen (N) and sulfur (S) associated with oil sands development in northern Alberta (Canada) has led to changing regional wet and dry N and S deposition regimes. We assessed the potential for using bog plant/lichen tissue chemistry (N and S concentrations, C:N and C:S ratios, in 10 plant/lichen species) to monitor changing atmospheric N and S deposition through sampling at five bog sites, 3-6 times per growing season from 2009 to 2016. During this 8-year period, oil sands N emissions steadily increased, while S emissions steadily decreased. We examined the following: (1) whether each species showed changes in tissue chemistry with increasing distance from the Syncrude and Suncor upgrader stacks (the two largest point sources of N and S emissions); (2) whether tissue chemistry changed over the 8 year period in ways that were consistent with increasing N and decreasing S emissions from oil sands facilities; and (3) whether tissue chemistry was correlated with growing season wet deposition of NH4+-N, NO3--N, or SO42--S. Based on these criteria, the best biomonitors of a changing N deposition regime were Evernia mesomorpha, Sphagnum fuscum, and Vaccinium oxycoccos. The best biomonitors of a changing S deposition regime were Evernia mesomorpha, Cladonia mitis, Sphagnum fuscum, Sphagnum capillifolium, Vaccinium oxycoccos, and Picea mariana. Changing N and S deposition regimes in the oil sands region appear to be influencing N and S cycling in what once were pristine ombrotrophic bogs, to the extent that these bogs may effectively monitor future spatial and temporal patterns of deposition.
Assuntos
Líquens , Áreas Alagadas , Alberta , Ascomicetos , Monitoramento Ambiental , Nitrogênio/análise , Campos de Petróleo e Gás , Parmeliaceae , Enxofre/análiseRESUMO
Bogs are nutrient poor, acidic ecosystems that receive their water and nutrients entirely from precipitation (= ombrogenous) and as a result are sensitive to nutrient loading from atmospheric sources. Bogs occur frequently on the northern Alberta landscape, estimated to cover 6% of the Athabasca Oil Sands Area. As a result of oil sand extraction and processing, emissions of nitrogen (N) and sulfur (S) to the atmosphere have led to increasing N and S deposition that have the potential to alter the structure and function of these traditionally nutrient-poor ecosystems. At present, no detailed protocol is available for monitoring potential change of these sensitive ecosystems. We propose a user-friendly protocol that will monitor potential plant and lichen responses to future environmental inputs of nutrients and provide a structured means for collecting annual data. The protocol centers on measurement of five key plant/lichen attributes, including changes in (1) plant abundances, (2) dominant shrub annual growth and primary production, (3) lichen health estimated through chlorophyll/phaeophytin concentrations, (4) Sphagnum annual growth and production, and (5) annual growth of the dominant tree species (Picea mariana). We placed five permanent plots in each of six bogs located at different distances from the center of oil sand extraction and sampled these for 2 years (2018 and 2019). We compared line intercept with point intercept plant assessments using NMDS ordination, concluding that both methods provide comparable data. These data indicated that each of our six bog sites differ in key species abundances. Structural differences were apparent for the six sites between years. These differences were mostly driven by changes in Vaccinium oxycoccos, not the dominant shrubs. We developed allometric growth equations for the dominant two shrubs (Rhododendron groenlandicum and Chamaedaphne calyculata). Equations developed for each of the six sites produced growth values that were not different from one another nor from one developed using data from all sites. Annual growth of R. groenlandicum differed between sites, but not years, whereas growth of C. calyculata differed between the 2 years with more growth in 2018 compared with 2019. In comparison, Sphagnum plant density and stem bulk density both had strong site differences, with stem mass density higher in 2019. When combined, annual production of S. fuscum was greater in 2019 at three sites and not different at three of the sites. Chlorophyll and phaeophytin concentrations from the epiphytic lichen Evernia mesomorpha also differed between sites and years. This protocol for field assessments of five key plant/lichen response variables indicated that both site and year are factors that must be accounted for in future assessments. A portion of the site variation was related to patterns of N and S deposition.
Assuntos
Nitrogênio , Áreas Alagadas , Alberta , Ecossistema , Monitoramento Ambiental , Nitrogênio/análise , Campos de Petróleo e GásRESUMO
Oil extraction and development activities in the Athabasca Oil Sands Region of northern Alberta, Canada, release NOx, SOx, and NHy to the atmosphere, ultimately resulting in increasing N and S inputs to surrounding ecosystems through atmospheric deposition. Peatlands are a major feature of the northern Alberta landscape, with bogs covering 6-10% of the land area, and fens covering 21-53%. Bulk deposition of NH4+-N, NO3--N, dissolved inorganic N (DIN), and SO42--S, was quantified using ion-exchange resin collectors deployed at 23 locations, over 1-6 years. The results reveal maximum N and S deposition of 9.3 and 12.0 kg ha-1 yr-1, respectively, near the oil sands industrial center (the midpoint between the Syncrude and Suncor upgrader stacks), decreasing with distance to a background deposition of 0.9 and 1.1 kg ha-1 yr-1, respectively. To assess potential influences of high N and S deposition on bogs, we quantified N and S concentrations in tissues of two Sphagnum species, two lichen species, and four vascular plant species, as well as surface porewater concentrations of H+, NH4+-N, NO3--N, SO42--S and dissolved organic N in 19 ombrotrophic bogs, distributed across a 3255 km2 sampling area surrounding the oil sands industrial center. The two lichen species (Evernia mesomorpha and Cladonia mitis), two vascular plant species (Rhododendron groenlandicum and Picea mariana), and to a lesser extent one moss (Sphagnum fuscum), showed patterns of tissue N and S concentrations that were (1) highest near the oil sands industrial center and (2) positively correlated with bulk deposition of N or S. Concentrations of porewater H+ and SO42--S, but not of NH4+-N, NO3--N, DIN, or dissolved inorganic N, also were higher near the oil sands industrial center than at more distant locations. The oil sands region of northern Alberta is remote, with few roads, posing challenges to the monitoring of oil sands-related N and S deposition. Quantification of N and S concentrations in bog plant/lichen tissues and porewaters may serve as a monitoring tool to assess both the local intensity and the spatial extent of bulk N and S deposition, and as harbingers of potential shifts in ecosystem structure and function.
Assuntos
Líquens , Áreas Alagadas , Alberta , Atmosfera/química , Monitoramento Ambiental , Campos de Petróleo e GásRESUMO
Bogs are ombrotrophic, relying solely on atmospheric deposition for new inputs of elements. Increased element deposition through anthropogenic activities has the potential to alter nutrient availability, and hence ecosystem function, in bogs. Further, because of efficient element retention, bogs may function as effective monitors of element deposition. To assess the potential effects of particulate fugitive dust from oil sands development in Alberta, Canada, we quantified plant/lichen tissue Ca, Mg, K, and P concentrations in 6 bogs ranging from 12 to 77 km from the oil sands industrial center. Deposition of Ca and Mg, but not K or P, quantified using ion exchange resin collectors, to bogs decreased with distance from the oil sands industrial center. Concentrations of Ca and Mg, but not K or P, in tissues of lichens (Cladonia mitis, Evernia mesomorpha) and Sphagnum (S. capillifolium, S. fuscum) decreased with distance from the oil sands industrial center. Tissue Ca concentrations were positively correlated with growing season Ca and Mg deposition in all species except Vaccinium oxycoccos, Rhododendron groenlandicum, and Picea mariana; leaf Mg concentrations were positively correlated with growing season Mg deposition for all species except P. mariana. Tissue concentrations of K and P were not correlated with growing season K and P deposition. For each species, receptor modeling identified two distinct sources, one dominated by Ca and Mg, presumed to represent particulate fugitive dust from oil sands activities, and a second dominated by K and P, which may reflect tight internal cycling and upward translocation of K and P in peat and/or K and P deposition as particulates generated in wildfires. Increasing Ca2+ and Mg2+ deposition may acidify bog porewaters through cation exchange in peat.
Assuntos
Líquens , Áreas Alagadas , Alberta , Poeira/análise , Ecossistema , Monitoramento Ambiental , Resinas de Troca Iônica , Campos de Petróleo e Gás , SoloRESUMO
Bogs and fens cover 6 and 21%, respectively, of the 140,329 km2 Oil Sands Administrative Area in northern Alberta. Regional background atmospheric N deposition is low (<2 kg N ha-1 yr-1), but oil sands development has led to increasing N deposition (as high as 17 kg N ha-1 yr-1). To examine responses to N deposition, over five years, we experimentally applied N (as NH4NO3) to a poor fen near Mariana Lake, Alberta, unaffected by oil sands activities, at rates of 0, 5, 10, 15, 20, and 25 kg N ha-1 yr-1, plus controls (no water or N addition). At Mariana Lake Poor Fen (MLPF), increasing N addition: 1) progressively inhibited N2-fixation; 2) had no effect on net primary production (NPP) of Sphagnum fuscum or S. angustifolium, while stimulating S. magellanicum NPP; 3) led to decreased abundance of S. fuscum and increased abundance of S. angustifolium, S. magellanicum, Andromeda polifolia, Vaccinium oxycoccos, and of vascular plants in general; 4) led to an increase in stem N concentrations in S. angustifolium and S. magellanicum, and an increase in leaf N concentrations in Chamaedaphne calyculata, Andromeda polifolia, and Vaccinium oxycoccos; 5) stimulated root biomass and production; 6) stimulated decomposition of cellulose, but not of Sphagnum or vascular plant litter; and 7) had no or minimal effects on net N mineralization in surface peat, NH4+-N, NO3--N or DON concentrations in surface porewater, or peat microbial composition. Increasing N addition led to a switch from new N inputs being taken up primarily by Sphagnum to being taken up primarily by shrubs. MLPF responses to increasing N addition did not exhibit threshold triggers, but rather began as soon as N additions increased. Considering all responses to N addition, we recommend a critical load for poor fens in Alberta of 3 kg N ha-1 yr-1.
Assuntos
Nitrogênio/análise , Sphagnopsida , Alberta , Campos de Petróleo e Gás , Solo , Áreas AlagadasRESUMO
Retroviral vectors infect primate bone marrow cells and express in vivo the transferred genes (the human ADA gene and the bacterial gene for neomycin resistance). The SAX vector appears to express human ADA at normal levels, but the infection efficiency is low (less than 1%) so that the gene product is only detectable in the peripheral blood at low levels. Vector expression disappears after 5 months (except for occasional T cells), presumably due to a failure to infect a renewal stem cell. While the level of ADA expression obtained in primates would not appear to be sufficient to correct outright the disease caused by ADA deficiency, it is possible that T-cell progenitors in the marrow will have a selective advantage. T cells expressing an ADA vector would then able to expand and potentially restore immune function. Unfortunately, this hypothesis will go untested until an animal model for ADA deficiency is found or a human clinical trial is performed. At present, consideration of gene therapy as a treatment for ADA deficiency would only be appropriate if all conventional forms of treatment were unsuccessful. If such a scenario should present itself, the critical question becomes one of safety, to both the patient and those in contact with the patient. We have begun to address the safety issues associated with gene therapy. Five animals exposed to replication-competent retrovirus during bone marrow transplantation show no evidence of helper virus, with a mean follow-up of 18.3 months. Four animals injected with replication-competent helper virus cleared the virus rapidly and, after the initial clearance, have shown no evidence of retroviremia, with a mean follow-up of 5.2 months. Our preliminary findings suggest that murine retorviruses do not cause a productive infection in vivo. These results, combined with the availability of better producer cell lines free of helper virus, are encouraging, and suggest that the risk of clinical disease from murine retrovirus introduced by a gene therapy protocol should be small. Unfortunately, high infection efficiency and long-term vector expression still must be obtained before retroviral-mediated gene transfer can be considered as first-line therapy for ADA deficiency.
Assuntos
Terapia Genética , Primatas/genética , Transfecção , Animais , Transplante de Medula Óssea , Vetores Genéticos , Humanos , Mutação , Retroviridae/genéticaRESUMO
Basic fibroblast growth factor (FGF-2) expression is associated with a more differentiated phenotype, earlier stage of disease, and a better prognosis in breast cancer patients. To determine whether expression of FGF-2 can cause a less malignant phenotype, we engineered MDA-MB-231 cells, a highly dedifferentiated, invasive breast cancer cell line, to express different isoforms of FGF-2. Cells expressed either cytoplasmic, nuclear, or a combination of both FGF-2 isoforms. Western blots of 2 M NaCl washes and of conditioned medium demonstrated that these cells did not export FGF-2. Cells expressing FGF-2 had levels of fibroblast growth factor receptors equivalent with those of control cells. Transformation was assayed by anchorage-independent colony formation and tumor formation in athymic mice. All of the constructs expressing various FGF-2 isoforms had a 60-70% reduction in colony formation in soft agar, but only cells expressing the Mr 18,000 FGF-2 isoform formed fewer and smaller tumors in mice. To determine potential mechanisms responsible for a less malignant phenotype, experiments measuring invasion in Matrigel, the secretion of matrix metalloprotease activity and migration in a modified Boyden chamber and in a patch wound motility assay were carried out. Cells expressing the Mr 18,000 cytoplasmic FGF-2 moiety had a 45% decrease in invasion in Matrigel compared to vector-transfected controls. Cells expressing Mr 18,000 FGF-2 had an increase in Mr 97,000 and Mr 48,000 collagenase, demonstrating that the decreased invasive potential was not due to a down-regulation of gelatinolytic or caseinolytic matrix metalloproteinases. However, motility was decreased in both assays, primarily in cells expressing Mr 18,000 FGF-2, whereas exogenous recombinant human FGF-2 had no effect. These studies demonstrate for the first time that FGF-2 expression can cause a less malignant phenotype in breast cancer cells, possibly as a result of decreased motility and invasion.
Assuntos
Neoplasias da Mama/patologia , Animais , Divisão Celular , Feminino , Humanos , Metaloproteinases da Matriz/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Invasividade Neoplásica , Fenótipo , Receptores de Fatores de Crescimento de Fibroblastos/fisiologia , TransfecçãoRESUMO
We investigated the capacity of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and all-trans-retinoic acid (ATRA) to sensitize three breast cancer cell lines to the cell killing effects of paclitaxel (Taxol) and Adriamycin, two chemotherapeutic agents commonly used in the treatment of breast cancer. In tissue culture colony assays, 1,25(OH)2D3 and ATRA were synergistic in inhibiting the clonogenicity of MCF-7 and T-47D cells that expressed estrogen receptor; vitamin D receptor; retinoic acid receptors (RARs) alpha, beta, and gamma; and retinoid X receptors alpha, beta, and gamma but were not additive in MDA-MB-231 cells that lacked expression of estrogen receptor, RARalpha, and RARbeta. The hormones used individually or in combination induced up to 40-50% cell death by a trypan blue exclusion assay in a dose-dependent manner up to concentrations of 10(-7) M in MCF-7 and T-47D cells, more modestly in MDA-MB-231 cells, and not at all in MCF-10 and MCF-12 nontransformed mammary epithelial cells. Pretreating the cancer cell lines with 1,25(OH)2D3 and ATRA individually or in combination for 3 days prior to a 1-h incubation with paclitaxel or Adriamycin decreased the ED50 for inhibition of colony formation or for cell death by trypan blue by up to 2 logs for paclitaxel and up to 1 log for Adriamycin in all three cell lines but had no effect on chemotherapy-induced MCF-12 cell death. The effects of the hormones were synergistic with those of the chemotherapy agents in all of the breast cancer cell lines, generally at the higher concentrations. Cell death took place by apoptosis. To determine one potential reason for the greater potentiation of the effects of paclitaxel than those of Adriamycin, we determined the effects of preincubation of MCF-7 cells on paclitaxel-induced phosphorylation of Bcl-2. Pretreatment of MCF-7 cells with either 1,25(OH)2D3 or ATRA increased the phosphorylation of Bcl-2 by variable concentrations of paclitaxel. These data suggest that pretreatment of breast cancer with 1,25(OH)2D3 or ATRA lowers the threshold for cell killing by chemotherapy agents and may provide a novel treatment option for this disease.
Assuntos
Antineoplásicos/toxicidade , Calcitriol/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/toxicidade , Paclitaxel/toxicidade , Tretinoína/toxicidade , Neoplasias da Mama , Sinergismo Farmacológico , Feminino , Humanos , Receptores de Calcitriol/fisiologia , Receptores de Estrogênio/fisiologia , Receptores do Ácido Retinoico/fisiologia , Receptores X de Retinoides , Fatores de Transcrição/fisiologia , Células Tumorais CultivadasRESUMO
Basic fibroblast growth factor (bFGF), a classical mitogen in fibroblasts and endothelial cells, inhibits the proliferation of MCF-7 and other human breast cancer cell lines. To explain this paradoxic effect, we investigated the effects of bFGF on cyclins and protein members of cyclin complexes that exert positive and negative control on the progression of cells through the G1 phase of the cell cycle. bFGF induced an increase in cyclin D1, cyclin E, and cyclin-dependent kinase 4 (cdk4) protein levels in a bFGF dose-dependent manner. However, bFGF also induced a heat-stable, transferable cytoplasmic factor in MCF-7 cells that inhibited the histone H1 kinase activity of reconstituted cyclin E-cdk2 and cyclin A-cdk2 complexes from Mv1Lu mink lung epithelial cells. The appearance of this inhibitor correlated with a bFGF dose- and time-dependent increase in the levels of cdk inhibitor p21(WAF1/CIP1) mRNA and protein. The increase in the level of p21(WAF1/CIP1) was associated with the disappearance of the rapidly migrating, activated form of cdk2 from cell lysates, dephosphorylation of the retinoblastoma protein (Rb), and a decrease in cyclin A levels. These changes were represented in the cyclin D1 and E complexes by an increased association with p21(WAF1/CIP1), proliferating cell nuclear antigen (PCNA), and the inactive form of cdk2, without an absolute change in cellular PCNA levels and by a switch in the association of cyclin D1 complexes with the hyperphosphorylated form to the dephosphorylated form of Rb. These experiments demonstrate that stimulation of MCF-7 cells with bFGF, although resulting in up-regulation of G1 proteins responsible for mitogenic events, also induces a concomitant decrease in cyclin A levels and an increase in p21(WAF1/CIP1) mRNA and protein and results in inactivation of cdk2, dephosphorylation of Rb, and a segregation of PCNA to the G1 cyclin complexes. The dual, conflicting signaling by bFGF results in a net inhibitory phenotype in these cells. These experiments suggest a pleiotropic role for bFGF in breast cancer.
Assuntos
Ciclo Celular/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Inibidores do Crescimento/farmacologia , Mitose/efeitos dos fármacos , Proteínas Proto-Oncogênicas , Western Blotting , Ciclina D1 , Quinase 4 Dependente de Ciclina , Inibidor de Quinase Dependente de Ciclina p21 , Quinases Ciclina-Dependentes/metabolismo , Ciclinas/genética , Ciclinas/metabolismo , DNA de Neoplasias/metabolismo , Feminino , Citometria de Fluxo , Humanos , Proteínas Oncogênicas/metabolismo , Proteínas Quinases/metabolismo , Fatores de Tempo , Células Tumorais CultivadasRESUMO
The effect of basic fibroblast growth factor (bFGF) on human breast cancer cells was studied in vitro. Exposure to bFGF resulted in significant growth inhibition, decreased DNA synthesis, and accumulation of cells in G0-G1. The IC50 for growth inhibition in MCF-7 cells was 50 pg/ml, and it was abrogated by neutralizing antibodies against bFGF. Inhibition of growth by bFGF was predominant over the growth stimulatory effects of 17beta-estradiol, insulin, or epidermal growth factor. Binding and cross-linking studies of 125I-labeled bFGF in intact MCF-7 cells demonstrated 5.2 x 10(3) saturable bFGF binding sites per cell, a dissociation constant of 57 pm, and a Mr 142,000 (125)I-labeled bFGF cross-linked protein. Stimulation of MCF-7 cells with bFGF at concentrations which effected growth inhibition also resulted in activation of p42(mapk) (ERK2) and p44(mapk) (ERK1) mitogen-activated protein kinases. These data demonstrate that whereas bFGF inhibits the growth of several breast cancer cell lines, it concomitantly activates ERK1 and ERK2, generally considered to signal mitogenic rather than growth inhibitory responses. Whether there is association between these phenomena remains unknown.
Assuntos
Neoplasias da Mama/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Fator 2 de Crescimento de Fibroblastos/farmacologia , Proteínas Quinases Ativadas por Mitógeno , Sítios de Ligação , Neoplasias da Mama/enzimologia , Ciclo Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , DNA/biossíntese , DNA/efeitos dos fármacos , Ativação Enzimática , Fator 2 de Crescimento de Fibroblastos/antagonistas & inibidores , Inibidores do Crescimento/farmacologia , Humanos , Radioisótopos do Iodo , Proteína Quinase 1 Ativada por Mitógeno , Proteína Quinase 3 Ativada por Mitógeno , Células Tumorais CultivadasRESUMO
Retroviral-mediated gene transfer into Rhesus monkey bone marrow cells, which were cryopreserved, stored, and then transduced at the time of thawing, was studied for potential application in gene therapy protocols. Albumin density gradient fractionation was used to define subpopulations of cryopreserved cells transduced by a murine retroviral vector. The transfer of the bacterial neomycin phosphotransferase gene into Rhesus monkey bone marrow that was cryopreserved and thawed was found to be preferential in a light-density population enriched for granulocyte-macrophage colony-forming units (CFU-GM). These results are similar to results obtained with freshly harvested bone marrow in which this population is enriched for CD34 antigen-positive cells, as well as for cells that were undergoing cell division. This method may be useful in enriching for transduced precursors in future gene transfer experiments in primates.
Assuntos
Células da Medula Óssea , Criopreservação , Terapia Genética/métodos , Vetores Genéticos , Transplante de Células-Tronco Hematopoéticas , Retroviridae , Preservação de Tecido , Transdução Genética , Animais , Antígenos CD/análise , Antígenos CD34 , Divisão Celular , Sobrevivência Celular , Células Cultivadas , Marcadores Genéticos , Granulócitos , Células-Tronco Hematopoéticas/imunologia , Células-Tronco Hematopoéticas/metabolismo , Canamicina Quinase , Macaca mulatta , Macrófagos , Camundongos , Fosfotransferases/biossíntese , Fosfotransferases/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genéticaRESUMO
We investigated cytosine arabinoside (Ara-C) as a potential agent for in vivo cycle activation of hematopoietic progenitors for the purpose of retroviral-mediated gene transfer. C57Bl mice were treated intraperitoneally with one of three regimens of Ara-C: a single 1,750 mg/kg dose (regimen 1), a 1,750 mg/kg dose on day 0, and a 1,500 mg/kg dose on day 2 (LD50) (regimen 2), or a 1,750 mg/kg dose on day 0 and a 1,500 mg/kg dose on day 3 (regimen 3). The high-proliferative-potential cells (HPPC)/10(5) cells were 47.0 +/- 7.5 pretreatment. The post-treatment HPPC cloning efficiencies were 40.6 +/- 3.4, 83.6 +/- 6.1, and 20.4 +/- 3.2 HPPC/10(5) cells on days 1, 2, and 4, respectively, with regimen 1; 60.0 +/- 7.9, 194.0 +/- 9.6, and 103.0 +/- 11.0 HPPC/10(5) cells 1, 2, and 4 days after the second Ara-C dose, respectively, with regimen 2; and 266 +/- 13.4, 132 +/- 23.9, and 118.0 +/- 5.7/10(5) cells 1, 2, and 4 days after the second Ara-C dose, respectively, with regimen 3. The transduction efficiency of HPPC from untreated animals with N2 viral supernatant was 4.9 +/- 5.8%.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Citarabina/farmacologia , Técnicas de Transferência de Genes , Células-Tronco Hematopoéticas/efeitos dos fármacos , Retroviridae/genética , Células 3T3 , Animais , Medula Óssea/efeitos dos fármacos , Células da Medula Óssea , Feminino , Vetores Genéticos , Hematopoese/efeitos dos fármacos , Humanos , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Basic fibroblast growth factor (bFGF) is a classical mitogen in fibroblasts and endothelial cells. Our previous studies have demonstrated that bFGF inhibits the growth of MCF-7 human breast cancer cells. The aim of the present study was to examine the effect of bFGF on cis-diamminedichloroplatinum(cisplatin)-induced cytotoxicity in MCF-7 breast cancer cells as compared to normal endothelial cells. MCF-7/NCF cells transduced with a vector expressing the bFGF gene and overexpressing its product, and MCF-7/N2 cells transduced with the backbone vector were incubated with a combination of bFGF and cisplatin for 5 days; results were compared with those obtained with bovine aortic endothelial cells. Cell proliferation was assessed with the sulforhodamine B colorimetric cytotoxicity assay. Apoptosis was quantitatively determined by flow-cytometric analysis for DNA damage and the apoptotic death assay for DNA fragmentation, and qualitatively by electron microscopy. Reverse transcriptase/polymerase chain reaction analysis and an enzyme immunoassay were used to determine the mRNA and protein level, respectively, of the anti-apoptotic bcl-2 gene product. We found that bFGF enhanced cisplatin-induced cytotoxicity in MCF-7 breast cancer sublines. bFGF enhanced proliferation of normal endothelial cells and did not increase cisplatin-induced cytotoxicity. This effect was accompanied by down-regulation of the anti-apoptotic protooncogene bcl-2 and the enhancement of cisplatin-induced apoptosis. We suggest that the improved understanding of the role of bFGF in the differential modulation of the response of breast cancer and normal endothelial cells to chemotherapy may enable active intervention to alter the therapeutic ratio favorably in breast cancer patients.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Cisplatino/farmacologia , Fator 2 de Crescimento de Fibroblastos/farmacologia , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Neoplasias da Mama/química , Neoplasias da Mama/patologia , Separação Celular , Cisplatino/uso terapêutico , DNA de Neoplasias/efeitos dos fármacos , Sinergismo Farmacológico , Feminino , Citometria de Fluxo , Humanos , Microscopia Eletrônica , Proteínas Proto-Oncogênicas c-bcl-2/análise , RNA Neoplásico/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais CultivadasRESUMO
Five leaking filtering blebs, occurring between 10 months and 21 years after trabeculectomy, were closed with cyanoacrylate tissue adhesive. Filtering bleb integrity was preserved in four cases, so that additional microsurgery was avoided. The only problem associated with use of tissue adhesive was the development of corneal abrasions in three cases. Although it is an accepted treatment for these conjunctival fistulas, only nine previous cases have been reported to our knowledge. We strongly recommend the use of tissue adhesive as a presurgical treatment in the management of leaking filtering blebs which present as a late postoperative complication of glaucoma surgery.
Assuntos
Cianoacrilatos , Complicações Pós-Operatórias/terapia , Trabeculectomia , Idoso , Túnica Conjuntiva , Doenças da Túnica Conjuntiva/terapia , Drenagem , Feminino , Fístula/terapia , Glaucoma/cirurgia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
A method for uniformly labeling kilogram amounts of plant litter with 14C is described. Data obtained from field decomposition of tagged fescue litter prepared by this method suggest that 14C flux can be a reliable indicator of energy flow, as measured directly by bomb calorimetry and indirectly by ashfree weight.