RESUMEN
The principal by-product from the two-phase olive oil production process is olive mill solid waste (OMSW). It is a highly-pollutant by-product, not only because of its characteristics, but also because of the considerable volume of OMSW which is generated, amounting to 2 to 4 million tons per year in Spain. The anaerobic digestion of this by-product is a well-studied process, and results in the generation of biogas, methane and carbon dioxide mainly of high calorific values (20-25 MJ m-3), and an effluent or digestate. The digestate of this by-product has never been characterized. This study presents an informative view on how the composition of OMSW digestate shows promising implications as a soil amendment or fertilizer due to the quality of the biomass from Lolium rigidum, a useful grass specie for the production of forage. Three OMSW digestate alternative applications or treatments were investigated: the digestate and the solid fraction of the digestate for a nutrient-poor soil amendment and the liquid fraction of the digestate as fertilizer. The results confirm that all the OMSW digestate treatments studied presented suitable characteristics for agricultural use, and showed an optimal Carbon/Nitrogen ratio with adequate values for heavy metals which are below the limits established by the Spanish and European legislation in the absence of pathogens. However, fertirrigation was the treatment that provided Lolium rigidum with the best characteristics, improving its shoot biomass, photosynthetic rate and nutritional content.
Asunto(s)
Lolium , Olea , Anaerobiosis , Fertilizantes , Poaceae , Suelo , Residuos SólidosRESUMEN
As the worldwide population continues to rise, so does global demand for agricultural production. This scenario of uncertain food supply is exacerbated by the high salinization of soils worldwide, a serious constraint to crop productivity. In this context, there is an increasing need for alternative sustainable crops. Halophytes are thought to be a promising alternative food source due to their natural ability to grow in saline soils and their multiple potential uses in the food industry. In this study, the protein and fatty acid content of the halophyte Halimione (Atriplex) portulacoides (L.) was studied in different saline conditions. Although more studies are needed to explore the nutritional properties of H. portulacoides, the data presented here suggest that this halophyte should be considered as a promising food crop for saline agriculture.
Asunto(s)
Chenopodiaceae/química , Ácidos Grasos/análisis , Proteínas de Plantas/análisis , Salinidad , Plantas Tolerantes a la Sal/química , Suelo/químicaRESUMEN
Spent livestock bedding is a valuable resource for the production of green energy (methane) in rural areas. Comparison and evaluation of batch anaerobic digestion and co-digestion of different mixtures of goat straw bedding (SGSB) and goat cheese whey were carried out. Biochemical methane potential (BMP) tests of the 100% SGSB, 95% SGSB-5% whey, 90% SGSB-10% whey, 85% SGSB-15% whey and 100% whey were found to be 423 ± 7, 354 ± 9, 371 ± 2, 293 ± 1, 274 ± 2 mL CH4 g-1 VS. Two different kinetic models were evaluated. The logistic model revealed a decrease in the maximum methane production rate (Rm) from 34.7 ± 1.5 to 14.1 ± 0.9 mL CH4 g-1 VS·d-1 when the percentage of whey in the mixture increased from 0 to 15% as a consequence of the increased ammonia released during the co-digestion of increased concentrations of whey. The lowest value for the maximum methane production predicted by the model (P) was found for 100% whey (274 ± 10 mL CH4 g-1 VS). A two-substrate model was applied to describe the evident existence of rapid and slowly degradable material. Regarding the hydrolysis kinetic constants predicted by this model, considerable increases in the rapid biodegradation stage (krapid) were observed when comparing to the values found for the slow (kslow) biodegradation stage in all the cases tested. The increases between both constants rose from 5 to 42% when the percentage of whey increased.
Asunto(s)
Queso , Suero Lácteo , Anaerobiosis , Animales , Ropa de Cama y Ropa Blanca , Biocombustibles/análisis , Reactores Biológicos , Digestión , Cabras , Metano/análisis , Suero Lácteo/químicaRESUMEN
Cherax quadricarinatus has been widely translocated within Australia, and a number of self-sustaining feral populations have established, and persisted, in central-eastern Australia for over 20 years: however, the biology and ecology of feral populations remain poorly understood. Using the loss-by-ignition method, this study investigated differences in the total content of incombustible material (as a proxy for total mineral content), between feral C. quadricarinatus populations in southeast Queensland and northeastern New South Wales. A total of 102 C. quadricarinatus were ignited, and percent total incombustible material was not proportional to the body size, or gender of the crayfish. Incombustible content was however, significantly different between some locations of capture (i.e., waterbodies). The site where incombustible content in crayfish was atypical, Lake Ainsworth, is a naturally acidic coastal lake, and we suggest that acidity and low concentration of calcium in that waterbody are likely responsible for the difference in mineral content detected in that population. Mechanism(s) driving the difference detected in the Lake Ainsworth population are unknown, but we suggest the acidic environment could directly impact maintenance of internal calcium reserves in the crayfish (intermoult), during recalcification of the cuticle (postmoult), or both. Limited calcium availability in the lake may also be a direct, or indirect, contributing factor. The ability of C. quadricarinatus to occupy acidic habitats while managing biomineralization challenges possibly could enable additional range-expansion of the species, and potential impacts on both endangered ecological communities and other biota occupying the acidic coastal habitats of Eastern Australia.
RESUMEN
Cholecystokinin-2 (CCK-2) receptors, overexpressed in cancer types such as small cell lung cancers (SCLC) and medullary thyroid carcinomas (MTC), may serve as targets for peptide receptor radionuclide imaging. A variety of CCK and gastrin analogues has been developed, but a major drawback is metabolic instability or high kidney uptake. The minigastrin analogue PP-F11 has previously been shown to be a promising peptide for imaging of CCK-2 receptor positive tumors and was therefore further evaluated. The peptide was conjugated with one of the macrocyclic chelators DOTA, NOTA, or NODAGA. The peptide conjugates were then radiolabeled with either (68)Ga, (64)Cu, or (111)In. All (radio)labeled compounds were evaluated in vitro (IC50) and in vivo (biodistribution and PET/CT and SPECT/CT imaging). IC50 values were in the low nanomolar range for all compounds (0.79-1.51 nM). In the biodistribution studies, (68)Ga- and (111)In-labeled peptides showed higher tumor-to-background ratios than the (64)Cu-labeled compounds. All tested radiolabeled compounds clearly visualized the CCK2 receptor positive tumor in PET or SPECT imaging. The chelator did not seem to affect in vivo behavior of the peptide for (111)In- and (68)Ga-labeled peptides. In contrast, the biodistribution of the (64)Cu-labeled peptides showed high uptake in the liver and in other organs, most likely caused by high blood levels, probably due to dissociation of (64)Cu from the chelator and subsequent transchelation to proteins. Based on the present study, (68)Ga-DOTA-PP-F11 might be a promising radiopharmaceutical for PET/CT imaging of CCK2 receptor expressing tumors such as MTC and SCLC. Clinical studies are warranted to investigate the potential of this tracer.
Asunto(s)
Acetatos/farmacología , Radioisótopos de Cobre/química , Radioisótopos de Galio/química , Gastrinas/química , Compuestos Heterocíclicos con 1 Anillo/farmacología , Compuestos Heterocíclicos/farmacología , Radioisótopos de Indio/química , Animales , Línea Celular Tumoral , Quelantes/química , Femenino , Humanos , Concentración 50 Inhibidora , Ratones , Ratones SCID , Imagen Multimodal , Trasplante de Neoplasias , Péptidos/química , Tomografía de Emisión de Positrones , Radiofármacos/química , Receptor de Colecistoquinina B/metabolismo , Tomografía Computarizada de Emisión de Fotón Único , Tomografía Computarizada por Rayos XRESUMEN
A glasshouse study of the coastal shrub Limoniastrum monopetalum was carried out to evaluate its tolerance and capacity to accumulate copper. We investigate the effects of Cu from 0 to 60 mmol l(-1) on the growth, photosynthetic apparatus, and nutrient uptake of L. monopetalum, by measuring gas exchange, chlorophyll fluorescence parameters, photosynthetic pigments, and total copper, nitrogen, phosphorus, sulfur, calcium, and magnesium content in the plant tissues. Although L. monopetalum did not survive at 60 mmol l(-1) Cu, the species demonstrated a high tolerance to Cu-induced stress, since all plants survived external Cu concentrations of up to 35 mmol l(-1) and displayed similar growth in the Cu-enriched medium as in the control treatment of up to the external level of 15 mmol Cu l(-1) (1,000 mg Cu l(-1)). The reduced growth registered in plants exposed to 35 mmol Cu l(-1) can be attributed to reduced photosynthetic carbon assimilation associated with the adverse effect of the metal on the photochemical apparatus and a reduction in the absorption of essential nutrients. Copper tolerance was associated with the capacity of the plant to accumulate the metal in its roots and effectively prevent its translocation to photosynthetic tissues. L. monopetalum has the characteristics of a Cu-excluder plant and could be used in the revegetation of Cu-contaminated soils.
Asunto(s)
Sulfato de Cobre/toxicidad , Fotosíntesis/efectos de los fármacos , Raíces de Plantas/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Tracheophyta/fisiología , Adaptación Fisiológica , Biodegradación Ambiental , Calcio/metabolismo , Magnesio/metabolismo , Nitrógeno/metabolismo , Fósforo/metabolismo , Fotosíntesis/fisiología , Raíces de Plantas/metabolismoRESUMEN
The present study evaluated the tolerance and accumulation potential in the salt-marsh shrub Halimione portulacoides under moderate and high external Cu levels. A greenhouse experiment was conducted in order to investigate the effects of a range of external Cu concentrations (0 to 60 mmol l(-1)) on growth and photosynthetic performance by measuring gas exchange, chlorophyll fluorescence parameters and photosynthetic pigments. We also determined total copper, nitrogen, phosphorus and sulfur concentrations in the plant tissues. H. portulacoides survived with external Cu concentrations of up to 35 mmol Cu l(-1), although the excess of metal resulted in a biomass reduction of 48%. The effects of Cu on growth were linked to a drastic reduction in net photosynthesis. However, H. portulacoides tolerated Cu levels of up to 15 mmol Cu l(-1) without suffering adverse physiological effects. Our results indicate that this species could play an important role in the restoration of Cu-contaminated soils.
Asunto(s)
Amaranthaceae/efectos de los fármacos , Amaranthaceae/metabolismo , Cobre/metabolismo , Cobre/toxicidad , Contaminantes Químicos del Agua/metabolismo , Contaminantes Químicos del Agua/toxicidad , Amaranthaceae/crecimiento & desarrollo , Restauración y Remediación Ambiental , Fotosíntesis/efectos de los fármacosRESUMEN
The halophytic shrub Halimione portulacoides is known to be capable of growth in soils containing extremely high concentrations of Zn. This study evaluated in detail the tolerance and accumulation potential of H. portulacoides under moderate and high external Zn levels. A greenhouse experiment was conducted in order to investigate the effects of a range of Zn concentrations (0-130 mmol L(-1)) on growth and photosynthetic performance by measuring relative growth rate, total leaf area, specific leaf area, gas exchange, chlorophyll fluorescence parameters and photosynthetic pigment concentrations. We also determined the total zinc, nitrogen, phosphorus, calcium, magnesium, sodium, potassium, iron and copper concentrations in the plant tissues. H. portulacoides demonstrated hypertolerance to Zn stress, since it survived with leaf concentrations of up to 2300 mg Zn kg(-1)dry mass, when treated with 130 mmol Zn L(-1). Zinc concentrations greater than 70 mmol L(-1) in the nutrient solution negatively affected plant growth, in all probability due to the recorded decline in net photosynthesis rate. Our results indicate that the Zn-induced decline in the photosynthetic function of H. portulacoides may be attributed to the adverse effect of the high concentration of the metal on photosynthetic electron transport. Growth parameters were virtually unaffected by leaf tissue concentrations as high as 1500 mg Zn kg(-1)dry mass, demonstrating the strong capability of H. portulacoides to protect itself against toxic Zn concentrations. The results of our study indicate that this salt-marsh shrub may represent a valuable tool in the restoration of Zn-polluted areas.
Asunto(s)
Amaranthaceae/metabolismo , Zinc/metabolismo , Amaranthaceae/fisiología , Calcio/metabolismo , Clorofila/metabolismo , Cobre/metabolismo , Hierro/metabolismo , Magnesio/metabolismo , Nitrógeno/metabolismo , Fósforo/metabolismo , Fotosíntesis , Potasio/metabolismo , Sodio/metabolismo , Suelo/química , HumedalesRESUMEN
The effects of vasoactive intestinal peptide (VIP) antagonists on breast cancer cells were investigated. (N-stearyl, norleucine17)VIP hybrid ((SN)VIPhyb) inhibited specific 125I-VIP binding to MCF7, SKBR3, T47D ZR75-1 and MDA-MB231 cells with high affinity (IC50 values of 0.03-0.06 microM). (SN)VIPhyb, 1 microM, inhibited the ability of 10 nM VIP to cause elevation of cAMP and to increase c-fos mRNA. Micromolar concentrations of (SN)VIPhyb inhibited the proliferation of MDA-MB231 or MCF7 cells using a MTT and clonogenic assay. Using a MTT assay, (SN)VIPhyb enhanced the ability of taxol and doxorubicin to inhibit breast cancer growth. Using nude mice bearing MDA-MB231 xenografts, VIPhyb potentiated the ability of taxol to inhibit proliferation. The results indicate that VIP receptor antagonists increase the ability of chemotherapeutic drugs to kill breast cancer cells.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Neurotensina/farmacología , Receptores de Péptido Intestinal Vasoactivo/antagonistas & inhibidores , Proteínas Recombinantes de Fusión/farmacología , Péptido Intestinal Vasoactivo/antagonistas & inhibidores , Péptido Intestinal Vasoactivo/farmacología , Secuencia de Aminoácidos , Animales , Antineoplásicos/uso terapéutico , División Celular/efectos de los fármacos , AMP Cíclico/metabolismo , Modelos Animales de Enfermedad , Doxorrubicina/farmacología , Sinergismo Farmacológico , Femenino , Genes fos/efectos de los fármacos , Humanos , Radioisótopos de Yodo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Datos de Secuencia Molecular , Neurotensina/uso terapéutico , Paclitaxel/farmacología , Unión Proteica/efectos de los fármacos , ARN Mensajero/efectos de los fármacos , Receptores de Péptido Intestinal Vasoactivo/metabolismo , Proteínas Recombinantes de Fusión/uso terapéutico , Timidina , Trasplante Heterólogo , Células Tumorales Cultivadas/efectos de los fármacos , Péptido Intestinal Vasoactivo/química , Péptido Intestinal Vasoactivo/metabolismo , Péptido Intestinal Vasoactivo/uso terapéuticoRESUMEN
The effects of indomethacin on A/J mice were investigated. The non-steroidal antiinflammatory drug (NSAID) indomethacin reduced significantly the number of lung adenomas 3, 4 or 8 months after urethane injection by 28, 30 and 29% respectively. The density of apoptotic cell bodies increased 2.9-fold in the lung adenomas of A/J mice treated with indomethacin. By immunocytochemistry, COX-2 immunoreactivity was present in the cytosol of lung adenomas, and in epithelial cells lining the bronchioli and bronchus as well as type 2 alveolar cells. COX-1 immunostaining was similar to that of COX-2 in the lungs of urethane-injected mice treated with or without indomethacin. By RT-PCR, COX-1 and COX-2 PCR products were present in mouse lung adenomas, alveoli and bronchioli. These results suggest that indomethacin may inhibit COX-1 and COX-2 in the A/J mouse lung resulting in reduced adenoma formation.
Asunto(s)
Adenoma/tratamiento farmacológico , Antiinflamatorios no Esteroideos/farmacología , Indometacina/farmacología , Neoplasias Pulmonares/tratamiento farmacológico , Adenoma/inducido químicamente , Animales , Apoptosis , Bronquios/metabolismo , Carcinógenos , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Citosol/metabolismo , Células Epiteliales/metabolismo , Femenino , Humanos , Inmunohistoquímica , Isoenzimas/biosíntesis , Neoplasias Pulmonares/inducido químicamente , Proteínas de la Membrana , Ratones , Prostaglandina-Endoperóxido Sintasas/biosíntesis , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Células Tumorales Cultivadas , UretanoRESUMEN
A synthetic peptide, (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) was used to investigate the signal transduction mechanisms of bombesin receptor subtype-3. Using NCI-1299#5 human lung cancer cells stably transfected with bombesin receptor subtype-3, 100 nM (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) elevated the cytosolic Ca2+ from 150 to 250 nM within 10 s. Addition of (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) caused phosphorylation of mitogen activated protein kinase in a time- and concentration-dependent manner. The mitogen activated protein kinase phosphorylation caused by (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) was inhibited by 2'-amino-3'-methyoxyflavone (PD98059), a mitogen activated protein kinase kinase (MEK-1) inhibitor. Using a luciferase reporter gene construct, (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) caused Elk-1 activation after 10 min and the increase in Elk-1 activation caused by (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) was inhibited by PD98059 as well as a dominant-negative MEK-1. (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) caused increased c-fos as well as c-jun mRNAs 1 h after addition to NCI-H1299#5 cells. The 47-fold increase in c-fos mRNA caused by 100 nM (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) was inhibited by PD98059, a dominant-negative MEK-1 and a substance P antagonist but not (3-phenylpropanoyl-D-Ala(24), Pro(26), Psi(26,27), Phe(27))GRP-(20-27) (BW2258U89), a GRP receptor antagonist. These results indicate that (D-Phe(6), beta-Ala(11), Phe(13), Nle(14))bombesin-(6-14) caused increased nuclear oncogene expression and upstream events include mitogen activated protein kinase phosphorylation and Elk-1 activation.
Asunto(s)
Bombesina/análogos & derivados , Proteínas de Unión al ADN , Inhibidores Enzimáticos/farmacología , Flavonoides/farmacología , Genes fos/efectos de los fármacos , Neoplasias Pulmonares/metabolismo , Quinasas de Proteína Quinasa Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Proto-Oncogénicas/efectos de los fármacos , ARN Mensajero/efectos de los fármacos , Receptores de Bombesina/efectos de los fármacos , Factores de Transcripción , Bombesina/farmacología , Calcio/metabolismo , Genes fos/fisiología , Humanos , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Oncogenes/efectos de los fármacos , Oncogenes/fisiología , Fragmentos de Péptidos/farmacología , Proteínas Proto-Oncogénicas/metabolismo , ARN Mensajero/metabolismo , Receptores de Bombesina/metabolismo , Células Tumorales Cultivadas , Proteína Elk-1 con Dominio etsRESUMEN
The effects of some oncogenes, growth factors and neuropeptides are mediated by tyrosine phosphorylation of focal adhesion kinase (p125(FAK)) and paxillin cytoskeletal proteins. In this study the ability of bombesin/gastrin releasing peptide (BB/GRP) to stimulate tyrosine phosphorylation of p125(FAK) and paxillin in non-small cell lung cancer (NSCLC) H1299 cells was investigated. BB, 100 nM caused increased p125(FAK) and paxillin tyrosine phosphorylation maximally after 1 min. The effect of BB on p125(FAK) and paxillin tyrosine phosphorylation was concentration-dependent, being half maximal at 4-8 nM. Also, 100 nM GRP, GRP(14-27) but not GRP(1-16) increased p125(FAK) and paxillin tyrosine phosphorylation indicating that the C-terminal of GRP is essential. BW2258U89, a GRP receptor antagonist, caused a dose-dependent inhibition of BB-stimulated p125(FAK) and paxillin tyrosine phosphorylation with an IC50 value of 3 microM. Cytochalasin D (0.3 microM), which inhibits actin polymerization, reduced the ability of BB to stimulate tyrosine phosphorylation of p125(FAK) and paxillin. Genistein (50 microM) and H-7 (50 microM), which are kinase inhibitors, reduced the tyrosine phosphorylation of p125(FAK) and paxillin stimulated by BB. Also, treatment of NCI-H1299 cells with FAK antisense resulted in decreased FAK tyrosine kinase activity and proliferation. These results suggest that p125(FAK) is an important enzyme for NSCLC proliferation.
Asunto(s)
Bombesina/farmacología , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Proteínas del Citoesqueleto/metabolismo , Péptido Liberador de Gastrina/farmacología , Neoplasias Pulmonares/metabolismo , Fosfoproteínas/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Tirosina/metabolismo , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/farmacología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/patología , División Celular/efectos de los fármacos , Quinasa 1 de Adhesión Focal , Proteína-Tirosina Quinasas de Adhesión Focal , Genisteína/farmacología , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/patología , Paxillin , Fosforilación , Células Tumorales CultivadasRESUMEN
The ability of nonpeptide antagonists to interact with neuromedin B receptors on C6 cells was investigated. 2-[3-(2, 6-Diisopropyl-phenyl)-ureido]3-(1H-indol-3-yl)-2-methyl-N-(1-pyridin- 2-yl-cyclohexylmethyl)-proprionate (PD165929), 3-(1H-indol-3-yl)-2-methyl-2-[3(4-nitro-phenyl)-ureido]-N-(1-pyridin- 2-yl-cyclohexylmethyl)-propionamide (PD168368) and 3-(1H-indol-3-yl)-N-[1-(5-methoxy-pyridin-2-yl)-cyclohexylmethyl]- 2-m ethyl-2-[3-(4-nitro-phenyl)-ureido]-propionamide (PD176252) inhibited (125I-Tyr0)neuromedin B binding with IC50 values of 2000, 40 and 50 nM, respectively. Because neuromedin B is a G-protein coupled serpentine receptor, the effects of neuromedin B antagonists on second messenger production and proliferation were investigated. PD168368 inhibited the ability of 10 nM neuromedin B to cause elevation of cytosolic Ca2+, whereas it had no effect on basal cytosolic Ca2+. PD168368 inhibited the ability of 100 nM neuromedin B to cause elevation of c-fos mRNA. Also, PD168368 in a dose-dependent manner inhibited the ability of 100 nM neuromedin B to cause phosphorylation of focal adhesion kinase. Using a [3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay, the order of antagonist potency to inhibit C6 proliferation was PD168368=PD176252>PD165929. Also, 1 microM PD168368 and PD176252 significantly inhibited colony number using a proliferation assay in vitro. PD168368 significantly inhibited C6 xenograft growth in nude mice in vivo. These results indicate that PD168368 is a C6 cell neuromedin B receptor antagonist, which inhibits proliferation.
Asunto(s)
División Celular/efectos de los fármacos , Neuroquinina B/análogos & derivados , Receptores de Bombesina/antagonistas & inhibidores , Animales , Unión Competitiva/efectos de los fármacos , Calcio/metabolismo , Células Clonales/citología , Células Clonales/efectos de los fármacos , Citosol/química , Citosol/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Indoles/farmacología , Ratones , Ratones Desnudos , Neuroquinina B/metabolismo , Neuroquinina B/farmacología , Proteínas Proto-Oncogénicas c-fos/genética , Piridinas/farmacología , ARN Mensajero/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Bombesina/metabolismo , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The effects of thymosin (THN) alpha1 were investigated using the urethane injection carcinogenesis A/J mouse model. Lung adenomas were observed 2.5, 3, and 4 months after urethane injection (400 mg/kg i.p.) into female A/J mice. Daily administration of THNalpha1 (0.4 mg/kg, s.c.) reduced lung adenoma multiplicity significantly, by approximately 45, 40, and 17%, respectively, 2.5, 3, and 4 months after urethane injection. Animals treated with THNalpha1 had a significantly greater white cell density than control A/J mice. Endogenous THNalpha1-like peptides were detected in the mouse lung. By radioimmunoassay and by Western blot, prothymosin alpha was detected in the mouse lung. By immunocytochemistry, THNalpha1-like peptides were detected in all lung compartments including the bronchus, adenoma, bronchioles, and alveoli. These results indicate that exogenous THNalpha1 prevents lung carcinogenesis in A/J mice.
Asunto(s)
Adenoma/prevención & control , Neoplasias Pulmonares/prevención & control , Timosina/análogos & derivados , Adenoma/inducido químicamente , Animales , Sangre/efectos de los fármacos , Western Blotting , Bronquios/metabolismo , Carcinógenos , Femenino , Inmunohistoquímica , Pulmón/efectos de los fármacos , Neoplasias Pulmonares/inducido químicamente , Ratones , Alveolos Pulmonares/metabolismo , Radioinmunoensayo , Timalfasina , Timosina/farmacología , Factores de Tiempo , Distribución Tisular , UretanoRESUMEN
The effects of sigma ligands on small cell lung cancer (SCLC) cells were investigated. 125I-N-(2-(piperidino)ethyl)-2-iodobenazmide (2-IBP) bound with high affinity to SCLC cell line NCI-H209 and NCI-N417. Specific 125I-2-IBP binding was inhibited with high affinity by ifendipine, haloperidol, (2-piperidinyl-aminoethyl)-4-iodobenzamide (IPAB) and 1,3-ditolylguanidine (DTG) with IC50 values of 3, 10, 15 and 90 nM respectively. In vitro, 10 microM 2-IBP, haloperidol or IPAB inhibited NCI-N417 proliferation using a MTT or clonogenic assay. In vivo, 4 mg/kg IPAB or 2-IBP inhibited NCI-N417 xenograft proliferation. 125I-2-IBP localized to the SCLC tumors after subcutaneous injection. These results suggest that sigma ligands may be utilized to localize and inhibit the proliferation of SCLC tumors.
Asunto(s)
Benzamidas/uso terapéutico , Carcinoma de Células Pequeñas/tratamiento farmacológico , División Celular/efectos de los fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Piperidinas/uso terapéutico , Receptores sigma/uso terapéutico , Animales , Benzamidas/metabolismo , Benzamidas/farmacocinética , Unión Competitiva/efectos de los fármacos , Carcinoma de Células Pequeñas/metabolismo , Carcinoma de Células Pequeñas/patología , Femenino , Guanidinas/farmacología , Haloperidol/farmacología , Humanos , Ligandos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Piperidinas/metabolismo , Piperidinas/farmacocinética , Receptores sigma/metabolismo , Células Tumorales CultivadasRESUMEN
The effects vasoactive intestinal peptide (VIP) antagonists were investigated on pancreatic cancer cell lines. (N-Stearyl, Norleucine17) VIP hybrid ((SN)VIPhyb) inhibited 125I-VIP binding to human Capan-2 cells with an IC50 value of 0.01 microM whereas VIP hybrid had an IC50 value of 0.2 microM. By RT-PCR and Northern blot, VPAC1 receptor mRNA was detected in CAPAN-2 cells. One microM (SN)VIPhyb and 10 microM VIPhyb inhibited the ability of 30 nM VIP to elevate cyclic AMP and increase c-fos mRNA. (SN)VIPhyb, 1 microM inhibited the clonal growth of CAPAN-2 cells in vitro. In vivo, (SN)VIPhyb (10 microg/day s.c.) inhibited CAPAN-2 xenograft growth in nude mice. These results indicate that (SN)VIPhyb is a pancreatic cancer VPAC receptor antagonist.
Asunto(s)
Neoplasias Pancreáticas/patología , Receptores de Péptido Intestinal Vasoactivo/antagonistas & inhibidores , Péptido Intestinal Vasoactivo/farmacología , Secuencia de Aminoácidos , Animales , Unión Competitiva , División Celular/efectos de los fármacos , AMP Cíclico/metabolismo , Genes fos/genética , Humanos , Concentración 50 Inhibidora , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Trasplante de Neoplasias , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/metabolismo , Unión Proteica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Péptido Intestinal Vasoactivo/genética , Receptores de Péptido Intestinal Vasoactivo/metabolismo , Receptores de Tipo I del Polipéptido Intestinal Vasoactivo , Trasplante Heterólogo , Células Tumorales Cultivadas , Péptido Intestinal Vasoactivo/antagonistas & inhibidores , Péptido Intestinal Vasoactivo/metabolismo , Péptido Intestinal Vasoactivo/uso terapéuticoRESUMEN
The effects of pituitary adenylate cyclase activating polypeptide (PACAP) analogs were investigated using breast cancer cells. 125I-PACAP-27 bound with high affinity (Kd = 5 nM) to T47D cells (Bmax = 29,000 per cell). Specific 125I-PACAP-27 binding was inhibited half maximally by PACAP-27, PACAP-38, PACAP(6-38) and PACAP(28-38) with IC50) values of 8, 17, 750 and >3000 nM, respectively. By RT-PCR, PACAP receptor mRNA was present in MCF-7 and T47D cell lines. Polyclonal antibodies to a PACAP receptor fragment (A-8-C) were elicited. The antibodies were affinity purified, recognized a 60-kDa protein by western blot, and stained malignant cells in breast cancer biopsy specimens by immunohistochemistry. PACAP-27 elevated the cAMP in T47D cells and the increase in cAMP caused by PACAP was inhibited by PACAP(6-38). PACAP-27 stimulated c-fos mRNA in T47D cells and the increase in c-fos gene expression caused by PACAP was reversed by PACAP(6-38). PACAP(6-38) inhibited colony formation using a soft agar assay and inhibited breast cancer xenograft growth in nude mice. These data suggest that PACAP(6-38) functions as a breast cancer PACAP receptor antagonist.
Asunto(s)
Neoplasias de la Mama/metabolismo , Neoplasias Mamarias Experimentales/metabolismo , Neuropéptidos/farmacología , Fragmentos de Péptidos/farmacología , Receptores de la Hormona Hipofisaria/antagonistas & inhibidores , Células 3T3 , Animales , Neoplasias de la Mama/enzimología , Neoplasias de la Mama/patología , División Celular/efectos de los fármacos , Femenino , Inhibidores de Crecimiento/metabolismo , Inhibidores de Crecimiento/farmacología , Humanos , Neoplasias Mamarias Experimentales/enzimología , Neoplasias Mamarias Experimentales/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Neuropéptidos/antagonistas & inhibidores , Neuropéptidos/metabolismo , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/metabolismo , Polipéptido Hipofisario Activador de la Adenilato-Ciclasa , Conejos , Receptores del Polipéptido Activador de la Adenilato-Ciclasa Hipofisaria , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
The effects of bombesin (BB) on mitogen activated protein (MAP) kinase were investigated using non-small cell lung cancer (NSCLC) cells. By Western blot, both 42 and 44 kDalton forms of MAP kinase were present in NCI-H1299 and NCI-H838 cells. Addition of BB to NCI-H1299 cells resulted in phosphorylation of the MAP kinase substrate myelin basic protein (MBP). Phosphorylation of MBP was maximal 6 min after the addition of 10 nM BB to NCI-H1299 cells. Addition of gastrin releasing peptide (GRP) or GRP14-27 but not GRP1-16 to NCI-H 1299 cells caused MBP phosphorylation. The effects of BB were inhibited by BW2258U89, a BB receptor antagonist, and PD98059, a MAP kinase kinase inhibitor. Also, PD98059 inhibited the clonal growth of NCI-H1299 cells. These data suggest that MAP kinase may be an important regulatory enzyme in NSCLC.
Asunto(s)
Bombesina/metabolismo , Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/enzimología , Neoplasias Pulmonares/enzimología , Secuencia de Aminoácidos , Carcinoma de Pulmón de Células no Pequeñas/patología , Péptido Liberador de Gastrina/farmacología , Humanos , Neoplasias Pulmonares/patología , Datos de Secuencia Molecular , Fosforilación/efectos de los fármacos , Células Tumorales CultivadasRESUMEN
The effects of lipoxygenase inhibitors were investigated using human lung cancer cell lines and A/J mice. By RT-PCR, 5-, 12-, and 15-lipoxygenase mRNA was detected in NSCLC cells. NDGA inhibited 5-LO activity in adenocarcinoma cell line NCI-H1264. Using an MTT assay, NDGA, MK591 and AA861 inhibited the growth of NSCLC cell lines tested with IC50 values of 3, 2, and 7 microM, respectively. Using a clonogenic assay, 10 microM NDGA significantly reduced NSCLC colony number. NDGA significantly slowed NSCLC xenograft growth in nude mice. When the tumors were excised and analyzed, nude mice treated with NDGA had significantly more apoptotic figures than did untreated tumors. A/J mice treated with urethane developed adenomas after 4 months and NDGA administration significantly reduced lung adenoma number. These data indicate that lipoxygenase inhibitors inhibit lung cancer growth and prevent lung carcinogenesis.