RESUMEN
BACKGROUND: Pronounced chemosensory adaptation affects many patients with olfactory loss. The study aimed to investigate adaptation to olfactory and trigeminal nasal stimuli in patients with olfactory loss in comparison to controls using electrophysiological measures. METHODOLOGY: Thirty-four patients with olfactory loss (mean age ± SD = 59 ± 16 years) and 17 healthy volunteers (mean age ± SD = 50 ± 14 years) were recruited. Sniffin♠sticks test was used for evaluation of olfactory function and EEG-derived chemosensory event-related potentials were recorded. Intranasal stimuli were presented using high-precision, computer-controlled stimulators based on the principles of air-dilution olfactometry. Data were analyzed in two different approaches according to the relatively short or long inter-stimulus interval. A decreased peak amplitude or a prolonged latency was considered as an expression of adaptation. RESULTS: The majority of participants (88%) responded reliably to chemosensory stimulation. Patients with olfactory loss exhibited pronounced olfactory and trigeminal adaptation within the long-term design, without such effects in healthy controls. Odor sensitivity correlated with both olfactory and trigeminal amplitude changes: the worse the olfactory sensitivity, the more pronounced chemosensory adaptation. CONCLUSIONS: The results help to explain the patients♠complaints in terms of the fast adaptation towards chemosensory stimuli, for example during eating and drinking. The differences in adaptation in patients with olfactory loss and healthy controls could serve as a clinical criterion to gauge olfactory dysfunction.
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Trastornos del Olfato , Olfato , Humanos , Olfato/fisiología , Potenciales Evocados/fisiología , Odorantes , Nervio TrigéminoRESUMEN
In recent years, there has been increasing use of the gap detection reflex test to demonstrate induction of tinnitus in animals. Animals with tinnitus show weakened gap detection ability for background noise that matches the pitch of the tinnitus. The usual explanation is that the tinnitus 'fills in the gap'. It has recently been shown, however, that tinnitus is commonly associated with hyperacusis-like enhancements of the acoustic startle response, a change which might potentially alter responses in the gap detection test. We hypothesized that such enhancements could lead to an apparent reduction of gap suppression, resembling that caused by tinnitus, by altering responses to the startle stimulus or the background noise. To test this hypothesis, we compared gap detection abilities in 3 subsets of noise-exposed animals with those in unexposed controls. The results showed that exposed animals demonstrated altered gap detection abilities, but these alterations were sometimes explained as consequences of hyper-responsiveness to either the startle stimulus or to the background noise. Two of the three subsets of animals studied, however, displayed weakened gap detection abilities that could not be explained by enhanced responses to these stimuli or by reduced sound sensitivity or a reduction of temporal processing speed, consistent with the induction of tinnitus. These results demonstrate that not only hearing loss but also changes in sensitivity to background noise or to startle stimuli are potential confounds that, when present, can underlie changes in gap detection irrespective of tinnitus. We discuss how such confounds can be recognized and how they can be avoided.
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Hiperacusia/etiología , Acúfeno/etiología , Acúfeno/fisiopatología , Estimulación Acústica/efectos adversos , Animales , Umbral Auditivo/fisiología , Conducta Animal , Cricetinae , Audición , Pérdida Auditiva/complicaciones , Ruido , Reflejo de Sobresalto/efectos de los fármacos , Filtrado Sensorial/fisiología , SonidoRESUMEN
Background Androgen deprivation therapy (ADT) in localized prostate cancer improves overall survival and is recommended by National Comprehensive Cancer Network guidelines in certain situations. However, ADT is without benefit in other situations and can actually cause harm. This study examines recent trends in the ADT use and quantifies the cost of guideline-discordant ADT. Patients and methods Patients, aged 66-80 years, in the Surveillance Epidemiology and End Results-Medicare database with non-metastatic prostate cancer diagnosed between 2004 and 2007 were included for analysis. Prostate-specific antigen, Gleason score, and stage were used to define D'Amico risk categories. Logistic regression was used to examine factors associated with guideline-discordant ADT. Annual direct cost was estimated using 2011 Medicare reimbursement for ADT. Results Of 28 654 men included, 12.4% received guideline-discordant ADT. In low-risk patients, 14.9% received discordant ADT, mostly due to simultaneous ADT with radiation. Discordant use was seen in 7.3% of intermediate and 14.9% of high-risk patients, mostly from ADT as primary therapy. The odds of receiving guideline-discordant ADT decreased over time (2007 versus 2004; OR 0.69; 95% CI 0.62-0.76). The estimated annual direct cost from discordant ADT is $42 000 000. Conclusion Approximately one in eight patients received ADT discordant with published guidelines. Elimination of discordant use would result in substantial savings.
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Antagonistas de Andrógenos/economía , Antagonistas de Andrógenos/uso terapéutico , Medicare/economía , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/economía , Anciano , Humanos , Masculino , Clasificación del Tumor , Estadificación de Neoplasias , Guías de Práctica Clínica como Asunto , Antígeno Prostático Específico/sangre , Programa de VERF , Estados UnidosRESUMEN
We report a case of moderate intrauterine growth delay with a congenital skull fracture and subdural hematoma, related to Menkes disease. The diagnosis was established in the neonatal period and absorptiometry showed global osteopenia. This disorder has an X-linked recessive inheritance pattern. It results from an abnormality in copper transport with a reduction in the ability to incorporate copper into certain enzymes that need it as a cofactor. The clinical phenotype stems from a deficiency of these enzymes, which explains the diversity of the symptoms. It begins in the first months of life with neurological disorders (hypotonia, seizures) and bone and vascular abnormalities. Usually, death occurs before the age of 5.
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Retardo del Crecimiento Fetal , Síndrome del Pelo Ensortijado , Factores de Edad , Enfermedades Óseas Metabólicas/diagnóstico , Encéfalo/diagnóstico por imagen , Resultado Fatal , Hematoma/diagnóstico , Humanos , Lactante , Recién Nacido , Imagen por Resonancia Magnética , Masculino , Síndrome del Pelo Ensortijado/diagnóstico , Síndrome del Pelo Ensortijado/diagnóstico por imagen , Cuero Cabelludo , Fracturas Craneales/diagnóstico por imagen , Tomografía Computarizada por Rayos XRESUMEN
BACKGROUND: Although it is uncommon, significant bleeding per rectum presents one of the most difficult emergency problems. Bleeding from a rectal ulcer is not well recognized as a cause of such bleeding. METHODS: From July 2000 through December 2000, 195 consecutive patients with significant blood loss per rectum were reviewed. RESULTS: Forty-eight cases in whom significant gastrointestinal (GI) bleeding occurred following prior hospitalization were identified. Sources of bleeding were gastroduodenal in 38 cases (79%) and colorectal in 10 cases (21%). The causes of inpatient colorectal bleeding were benign rectal ulcer (n = 4), ischemic colitis (n = 3), neoplasia (n = 2), and diversion colitis (n = 1). CONCLUSION: The differential diagnosis for inpatients who develop new inpatient GI bleeding differs from that of patients who develop outpatient GI bleeding. Careful examination of the rectum following rectal instrumentation is critical. In addition to the standard resuscitative measures, the identification and treatment of rectal ulcers in this group of patients is of paramount importance. The treatment options for bleeding rectal ulcer include conservative therapy, cauterization, embolization, banding, and local excision.
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Hemorragia Gastrointestinal/etiología , Enfermedades del Recto/complicaciones , Úlcera/complicaciones , Adulto , Anciano , Anciano de 80 o más Años , Algoritmos , Transfusión Sanguínea , Colitis Isquémica/complicaciones , Comorbilidad , Diagnóstico Diferencial , Divertículo/complicaciones , Embolización Terapéutica , Urgencias Médicas , Femenino , Hemorragia Gastrointestinal/diagnóstico , Humanos , Pacientes Internos , Ligadura , Masculino , Persona de Mediana Edad , Pacientes Ambulatorios , Complicaciones Posoperatorias/diagnóstico , Enfermedades del Recto/diagnóstico , Enfermedades del Recto/cirugía , Enfermedades del Recto/terapia , Estudios Retrospectivos , Úlcera/diagnóstico , Úlcera/cirugía , Úlcera/terapiaRESUMEN
We examined the interaction between Alimta and ionizing radiation (IR) as a potential strategy to enhance the therapeutic ratio of combined-modality cancer treatment. Mice bearing human esophageal adenocarcinoma xenografts (Seg-1) or squamous cell carcinoma xenografts (SQ-20B) were treated with Alimta and IR employing a fractionated treatment schedule. Treatment with Alimta alone slowed the growth of Seg-1 but not SQ-20B tumors compared with control tumors. In Seg-1 xenografts combined treatment with Alimta and IR produced significant tumor growth inhibition compared with Alimta alone or IR alone. In SQ-20B xenografts, treatment with Alimta did not enhance IR-mediated tumor growth inhibition suggesting that sensitivity to Alimta is necessary for an interactive cytotoxic effect with IR. The present data suggest the potential clinical efficacy of combining Alimta administration with radiotherapy for Alimta-sensitive cells and indicate that further testing needs to be conducted to optimize the dosing schedule to enhance the interaction between the therapeutic agents.
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Antineoplásicos/uso terapéutico , Antagonistas del Ácido Fólico/uso terapéutico , Glutamatos/uso terapéutico , Guanina/análogos & derivados , Guanina/uso terapéutico , Neoplasias/tratamiento farmacológico , Neoplasias/radioterapia , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/patología , Adenocarcinoma/radioterapia , Animales , Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/radioterapia , Terapia Combinada , Relación Dosis-Respuesta en la Radiación , Neoplasias Esofágicas/tratamiento farmacológico , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/radioterapia , Femenino , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Neoplasias de Cabeza y Cuello/patología , Neoplasias de Cabeza y Cuello/radioterapia , Humanos , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias/patología , Pemetrexed , Radiación Ionizante , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
PURPOSE: The purpose of this study was to evaluate whether endostatin, an antiangiogenic cleavage fragment of collagen XVIII, enhances the antitumor effects of ionizing radiation (IR). Endostatin was injected to coincide with fractionated radiotherapy. METHODS: Xenografts of radioresistant SQ-20B tumor cells were established in athymic nude mice. Lewis lung carcinoma cells were injected into C57BI/6 mice. Mice bearing SQ-20B xenografts were injected intraperitoneally with 2.5 mg/kg/day of murine recombinant endostatin 5 times per week for 2 weeks 3 hours before IR treatment (50 Gy total dose). Mice bearing Lewis lung carcinoma tumors were injected intraperitoneally with endostatin (2.5 mg/kg/day) four times; the first injection was given 24 hours before the first IR dose (15 Gy) and then 3 hours before IR (15 Gy/day) for 3 consecutive days. Microvascular density was assessed on tumor tissue sections by use of CD31 immunohistochemistry and light microscopy. Endothelial cell survival analyses were employed to evaluate endostatin effects on human aortic endothelial cells and human umbilical vein endothelial cells. Endothelial cell apoptosis was examined by use of FACS analysis and DAPI microscopy. RESULTS: In SQ-20B xenografts, combined treatment with endostatin and IR produced tumor growth inhibition that was most pronounced at the nadir of regression (day 21). By day 35, tumors receiving combined treatment with endostatin and IR were 47% smaller than tumors treated with endostatin alone. Interactive cytotoxic treatment effects between endostatin and IR were also demonstrated in mice bearing Lewis lung carcinoma tumors. Significant tumor growth inhibition was observed in the endostatin/IR group at days 11 and 13 compared with IR alone. Histologic analyses demonstrated a reduction in microvascular density after combined treatment with endostatin and IR compared with endostatin treatment alone. Survival analyses confirmed interactive cytotoxicity between endostatin and IR in both human aortic endothelial cells and human umbilical vein endothelial cells but not in SQ-20B tumor cells. Combined treatment with endostatin and IR produced an increase in cow pulmonary artery endothelial apoptosis compared with either treatment alone. DISCUSSION: The tumor regression observed after combined treatment with endostatin and IR suggests additive antitumor effects in both human and murine tumors. Importantly, the concentrations of endostatin employed produced little tumor regression when endostatin was employed as a single agent. The results from the clonogenic and apoptosis assays support the hypothesis that the endothelial compartment is the target for the endostatin/IR interaction.
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Antineoplásicos/uso terapéutico , Colágeno/uso terapéutico , Neoplasias/tratamiento farmacológico , Neoplasias/radioterapia , Fragmentos de Péptidos/uso terapéutico , Radiación Ionizante , Animales , Apoptosis , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Separación Celular , Células Cultivadas , Clonación Molecular , Colágeno Tipo XVIII , Terapia Combinada , Relación Dosis-Respuesta a Droga , Endostatinas , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Escherichia coli/metabolismo , Femenino , Citometría de Flujo , Humanos , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Microcirculación/efectos de los fármacos , Microcirculación/efectos de la radiación , Trasplante de Neoplasias , Neoplasias/metabolismo , Pichia/metabolismo , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/biosíntesis , Proteínas Recombinantes/metabolismo , Factores de Tiempo , Células Tumorales Cultivadas , Venas Umbilicales/citología , Venas Umbilicales/efectos de los fármacosRESUMEN
BACKGROUND: Although ionizing radiation (IR) has been demonstrated to attenuate vessel wall restenosis and intimal hyperplasia (IH), dose-related mural injury and atrophy are possible deleterious side effects. We tested the hypothesis that a radiosensitizing strategy may improve IR-induced inhibition of in vivo vascular smooth muscle cells (VSMCs) without influencing apoptotic cell death. METHODS: In 28 New Zealand White rabbits, the right common carotid artery (CCA) was injured and subjected to low-flow conditions to promote IH. The CCA was transfected with an adenoviral vector incorporating the cytosine deaminase (CD) gene (1 x 10(9) PFU/ml). 5-Fluorocytosine (5-FC), a prodrug that is converted to the radiosensitizing agent 5-fluorouracil (5-FU) by CD, was thereafter administered intravenously. The CCA was exposed to 5 Gy IR at 24 h. Intimal/medial (I/M) area and thickness ratios were determined in the harvested CCAs at 14 days. VSMC proliferative and apoptotic indices were assessed with immunohistochemistry. RESULTS: A 50% reduction in I/M area was found in rabbits treated with IR and IR + CD/5-FC (0.19 +/- 0.03 and 0.18 +/- 0.02) when compared with untreated controls (UC) (0.37 +/- 0.06) (P = 0.005). This finding was substantiated by attenuation of I/M thickness in the IR groups [0.47 +/- 0.13 (IR), 0.41 +/- 0.11 (IR + CD/5-FC), 0.61 +/- 0.17 (UC)] (P = 0.007). The number of proliferating VSMCs was notably smaller when IR was combined with CD/5-FC (4.17 +/- 1.16 vs 2.97 +/- 1.09 log transformed cells/mm(2), P < 0.07). Apoptosis was similar in all groups. CONCLUSIONS: Both IR alone and IR combined with a radiosensitizing agent are effective in attenuating experimental IH. However, combination therapy is synergistic and achieves greater inhibition of VSMC proliferation and may involve selective killing of radioresistant S-phase VSMCs. IR + CD/5-FC represents a novel therapeutic strategy that offers potential for long-term control of IH.
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Terapia Genética , Túnica Íntima/patología , Túnica Íntima/efectos de la radiación , Animales , Apoptosis/efectos de los fármacos , Apoptosis/efectos de la radiación , Arteria Carótida Común/efectos de los fármacos , Arteria Carótida Común/patología , Arteria Carótida Común/fisiopatología , División Celular/efectos de los fármacos , División Celular/efectos de la radiación , Citosina Desaminasa , Flucitosina/farmacología , Hemodinámica/efectos de los fármacos , Hemodinámica/efectos de la radiación , Hiperplasia/patología , Masculino , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/patología , Músculo Liso Vascular/fisiopatología , Nucleósido Desaminasas/genética , Profármacos/farmacología , Conejos , Túnica Íntima/efectos de los fármacosRESUMEN
We examined the effects of a new antiangiogenic isocoumarin, NM-3, as a radiation modifier in vitro and in vivo. The present studies demonstrate that NM-3 is cytotoxic to human umbilical vein endothelial cells (HUVECs) but not to Lewis lung carcinoma (LLC) cells nor Seg-1, esophageal adenocarcinoma cells, in clonogenic survival assays. When HUVEC cultures are treated with NM-3 combined with ionizing radiation (IR), additive cytotoxicity is observed. In addition, the combination of NM-3 and IR inhibits HUVEC migration to a greater extent than either treatment alone. The effects of treatment with NM-3 and IR were also evaluated in tumor model systems. C57BL/6 female mice bearing LLC tumors were given injections for 4 consecutive days with NM-3 (25 mg/kg/day) and treated with IR (20 Gy) for 2 consecutive days. Combined treatment with NM-3 and IR significantly reduced mean tumor volume compared with either treatment alone. An increase in local tumor control was also observed in LLC tumors in mice receiving NM-3/IR therapy. When athymic nude mice bearing Seg-1 tumor xenografts were treated with NM-3 (100 mg/kg/day for 4 days) and 20 Gy (four 5 Gy fractions), significant tumor regression was observed after combined treatment (NM-3 and IR) compared with IR alone. Importantly, no increase in systemic or local tissue toxicity was observed after combined treatment (NM-3 and IR) when compared with IR alone. The bioavailability and nontoxic profile of NM-3 suggests that the efficacy of this agent should be tested in clinical radiotherapy.
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Cumarinas/farmacología , Neoplasias/tratamiento farmacológico , Neoplasias/radioterapia , Adenocarcinoma/tratamiento farmacológico , Animales , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Movimiento Celular/efectos de los fármacos , Movimiento Celular/efectos de la radiación , Células Cultivadas , Colágeno/metabolismo , Cumarinas/toxicidad , Relación Dosis-Respuesta a Droga , Relación Dosis-Respuesta en la Radiación , Combinación de Medicamentos , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/efectos de la radiación , Neoplasias Esofágicas/tratamiento farmacológico , Femenino , Humanos , Isocumarinas , Laminina/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Desnudos , Trasplante de Neoplasias , Proteoglicanos/metabolismo , Radiación Ionizante , Factores de Tiempo , Células Tumorales Cultivadas , Venas Umbilicales/citología , Venas Umbilicales/efectos de los fármacos , Venas Umbilicales/efectos de la radiaciónRESUMEN
The family of vascular endothelial growth factor (VEGF) proteins include potent and specific mitogens for vascular endothelial cells that function in the lation of angiogenesis Inhibition of VEGF-induced angiogenesis either by neutralizing antibodies or dominant-negative soluble receptor, blocks the growth of primary and metastatic experimental tumors Here we report that VEGF expression is induced in Lewis lung carcinomas (LLCs) both in vitro and vivo after exposure to ionizing radiation (IR) and in human tumor cell lines (Seg-1 esophageal adenocarcinoma, SQ20B squamous cell carcinoma, T98 and U87 glioblastomas, and U1 melanoma) in vitro. The biological significance of IR-induced VEGF production is supported by our finding that treatment of tumor-bearing mice (LLC, Seg-1, SQ20B, and U87) with a neutralizing antibody to VEGF-165 before irradiation is associated with a greater than additive antitumor effect. In vitro, the addition of VEGF decreases IR-induced killing of human umbilical vein endothelial cells, and the anti-VEGF treatment potentiates IR-induced lethality of human umbilical vein endothelial cells. Neither recombinant VEGF protein nor neutralizing antibody to VEGF affects the radiosensitivity of tumor cells These findings support a model in which induction of VEGF by IR contributes to the protection of tumor blood vessels from radiation-mediated cytotoxicity and thereby to tumor radioresistance.
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Anticuerpos Monoclonales/farmacología , Factores de Crecimiento Endotelial/antagonistas & inhibidores , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Linfocinas/antagonistas & inhibidores , Proteínas de Neoplasias/antagonistas & inhibidores , Neoplasias Experimentales/radioterapia , Neovascularización Patológica/fisiopatología , Fármacos Sensibilizantes a Radiaciones/farmacología , Estrés Fisiológico/fisiopatología , Adenocarcinoma/genética , Adenocarcinoma/patología , Adenocarcinoma/radioterapia , Animales , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/radioterapia , Células Cultivadas , Medios de Cultivo Condicionados , Factores de Crecimiento Endotelial/inmunología , Factores de Crecimiento Endotelial/fisiología , Endotelio Vascular/citología , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/radioterapia , Femenino , Glioblastoma/genética , Glioblastoma/patología , Glioblastoma/radioterapia , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Linfocinas/inmunología , Linfocinas/fisiología , Melanoma/genética , Melanoma/patología , Melanoma/radioterapia , Ratones , Ratones Endogámicos C57BL , Ratones Desnudos , Proteínas de Neoplasias/inmunología , Proteínas de Neoplasias/fisiología , Trasplante de Neoplasias , Neoplasias Experimentales/irrigación sanguínea , Neoplasias Experimentales/complicaciones , Neoplasias Experimentales/fisiopatología , ARN Mensajero/biosíntesis , ARN Neoplásico/biosíntesis , Tolerancia a Radiación/efectos de los fármacos , Estrés Fisiológico/genética , Células Tumorales Cultivadas/metabolismo , Células Tumorales Cultivadas/efectos de la radiación , Ensayo de Tumor de Célula Madre , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
Angiostatin, a proteolytic fragment of plasminogen, inhibits the growth of primary and metastatic tumors by suppressing angiogenesis. When used in combination with ionizing radiation (IR), angiostatin demonstrates potent antitumor synergism, largely caused by inhibition of the tumor microvasculature. We report here the temporal interaction of angiostatin and IR in Lewis lung carcinoma (LLC) tumors growing in the hind limbs of syngeneic mice. Tumors with an initial mean volume of 510 +/- 151 mm3 were treated with IR alone (20 Gy x 2 doses on days 0 and 1), angiostatin alone (25 mg/kg/day divided twice daily) on days 0 through 13, or a combination of the two as follows: (a) IR plus angiostatin (days 0 through 13); (b) IR plus angiostatin (days 0 and 1); and (c) IR followed by angiostatin beginning on the day after IR completion and given daily thereafter (days 2 through 13). By day 14, tumors in untreated control mice had grown to 6110 +/- 582 mm3, whereas in mice treated with: (a) IR alone, tumors had grown to 2854 +/- 338 mm3 (P < 0.05 compared with untreated controls); and (b) angiostatin alone, tumors had grown to 3666 +/- 453 mm3 (P < 0.05 compared with untreated controls). In combined-treatment groups, in mice treated with: (a) IR plus longer-course angiostatin, tumors reached 2022 +/- 282 mm3 (P = 0.036 compared with IR alone); (b) IR followed by angiostatin, tumors reached 2677 +/- 469 mm3 (P > 0.05 compared with IR alone); and (c) IR plus short-course angiostatin, tumors reached 1032 +/- 78 mm3 (P < 0.001 compared with IR alone). These findings demonstrate that the efficacy of experimental radiation therapy is potentiated by brief concomitant exposure of the tumor vasculature to angiostatin.
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Antineoplásicos/uso terapéutico , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Carcinoma Pulmonar de Lewis/radioterapia , Fragmentos de Péptidos/uso terapéutico , Plasminógeno/uso terapéutico , Angiostatinas , Animales , Terapia Combinada , Esquema de Medicación , Femenino , Ratones , Ratones Endogámicos C57BL , Fragmentos de Péptidos/administración & dosificación , Plasminógeno/administración & dosificación , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
BACKGROUND: Epidermal growth factor (EGF) stimulates cell replication and increases DNA content of the small intestine, but its effects on mucosal amino acid transport are unknown. METHODS: To investigate these effects, we treated adult rats with vehicle or EGF (10 micrograms/100 gm body weight subcutaneously every 8 hours for three doses). Jejunal brush border membrane vesicles (BBMVs) from each group were prepared by Mg++ aggregation/differential centrifugation. BBMVs were enriched fifteen-fold in alkaline phosphatase, indicating BBMV purity. Transport of 3H-glutamine and 3H-alanine was studied by a rapid mixing filtration technique. Uptakes were primarily Na+ dependent, occurred in an osmotically active space, exhibited classic overshoots, and had similar 2-hour equilibrium values. RESULTS: Glutamine transport by BBMVs more than doubled in rats treated with EGF (16.4 +/- 0.1 pmol glutamine/mg protein/10 sec in EGF vs 7.1 +/- 0.5 pmol glutamine/mg protein/10 sec in controls; p < 0.001). Kinetic studies of the glutamine transporter showed that the increase in transport was the result of a 70% increase in maximal transport velocity (total maximum glutamine uptake = 193 +/- 8 pmol glutamine/mg protein/10 sec in EGF vs 114 +/- 7 pmol glutamine/mg protein/10 sec in controls; p < 0.0001 with no change in transporter affinity (transporter affinity = 224 +/- 6 mumol/L in EGF vs 242 +/- 37 mumol/L in controls; difference, not significant). Alanine uptake by BBMVs was also increased with EGF administration (10.2 +/- 2.0 pmol alanine/mg protein/10 sec in EGF vs 4.5 +/- 0.5 pmol alanine/mg protein/10 sec in controls; p < 0.005). Simultaneously, glucose transport was decreased by 50% in EGF-treated rats, indicating that the Na(+)-dependent glucose cotransporter is regulated independently from and opposite to amino acid transporters. CONCLUSIONS: We conclude that EGF up-regulates amino acid transport activity in jejunal BBMVs, an event that is most likely caused by an increase in de novo biosynthesis of transporter protein. The increase in amino acid uptake not only may support de novo protein synthesis but, in the case of glutamine, also may be required for energy production and nucleotide biosynthesis.
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Factor de Crecimiento Epidérmico/farmacología , Glutamina/metabolismo , Intestino Delgado/metabolismo , Alanina/metabolismo , Animales , Transporte Biológico , División Celular , ADN/metabolismo , Yeyuno/metabolismo , Cinética , Masculino , Microvellosidades/metabolismo , Ratas , Ratas Sprague-Dawley , Sodio/farmacologíaRESUMEN
The sodium-dependent transporter system responsible for L-glutamine uptake by brush border membrane vesicles prepared from equine jejunum was characterized. Vesicle purity was ascertained by a 14- to 17-fold increase in activity of the brush border enzyme markers. Glutamine uptake was found to occur into an osmotically active space with negligible membrane binding. The sodium-dependent velocity represented approximately 80% of total uptake and demonstrated overshoots. Kinetic studies of sodium-dependent glutamine transport at concentrations between 5 microM and 5 mM revealed a single saturable high-affinity carrier with a Michaelis constant of 519 +/- 90 microM and a maximal transport velocity of 3.08 +/- 0.97 nmol/mg of protein/10 s. Glutamine uptake was not affected by changes in environmental pH. Lithium could not substitute for sodium as a contransporter ion. 2-Methylaminoisobutyric acid inhibited the sodium-dependent carrier only minimally, but marked inhibition (> 90%) was observed in the presence of histidine, alanine, cysteine, and nonradioactive glutamine. Kinetic analysis of the sodium-independent transporter revealed it to have a Michaelis constant = 260 +/- 47 microM and a maximal transport velocity of 0.32 +/- 0.06 nmol/mg of protein/10 s. We conclude that glutamine transport in equine jejunal brush border membrane vesicles occurs primarily via the system B transporter and, to a lesser extent, by a sodium-independent carrier.
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Glutamina/farmacocinética , Caballos/metabolismo , Yeyuno/metabolismo , Aminoácidos/farmacología , Animales , Transporte Biológico/efectos de los fármacos , Transporte Biológico/fisiología , Cationes/farmacología , Concentración de Iones de Hidrógeno , Técnicas In Vitro , Absorción Intestinal/efectos de los fármacos , Absorción Intestinal/fisiología , Microvellosidades/metabolismo , Modelos Biológicos , Potasio/fisiología , Sodio/fisiologíaRESUMEN
The early effects of endotoxin (4 hr after a single dose of Escherichia coli LPS, 7.5 mg/kg) on L-glutamine (GLN) transport across the jejunal brush border of rats were studied. Jejunal brush border membrane vesicles (BBMVs) were prepared by a Mg2+ aggregation/differential centrifugation technique. Vesicle purity and integrity were confirmed by a 15-fold enrichment of brush border marker enzymes, osmotic activity, transport overshoots in the presence of sodium, and similar 1- and 2-hr equilibrium values. L-[3H]GLN transport in jejunal BBMVs was measured by a millipore filtration technique. Na(+)-dependent glutamine transport, which accounted for greater than 80% of total transport, was increased twofold in BBMVs from endotoxin-treated rats (67 +/- 5 pmole/mg protein/15 sec vs 38 +/- 3, P less than 0.01). Endotoxin treatment did not alter the activity of the Na(+)-independent carrier. Simultaneously, intestinal extraction of glutamine from the bloodstream fell by 56% (15.1 +/- 2.3% in controls vs 6.6 +/- 1.3% in endotoxin-treated rats, P less than 0.01). This reduction in the uptake of circulating glutamine could not be accounted for by a fall in the arterial concentration. Thus, soon after endotoxemia brush border glutamine uptake is increased while consumption of glutamine across the basolateral membrane is decreased. This increased uptake may support protein synthesis and may provide a biochemical rationale for the use of early enteral nutrition after the onset of critical illness.
Asunto(s)
Proteínas Portadoras/metabolismo , Endotoxinas/sangre , Escherichia coli , Yeyuno/metabolismo , Animales , Transporte Biológico , Glutamina/metabolismo , Mucosa Intestinal/metabolismo , Masculino , Microvellosidades/metabolismo , Ratas , Ratas EndogámicasRESUMEN
The effects of tumor necrosis factor and interleukin-1 on sodium-dependent glutamine transport by cultured pulmonary artery endothelial cells (PAECs) were studied. Incubation of PAECs with cytokines (10 to 1000 units/ml) resulted in a significant increase in System ASC-mediated glutamine transport that was dose-dependent, first observable after 8 hours, and maximal after 12 hours of exposure. Kinetic studies indicated that the increase in carrier-mediated activity was not due to a change in Km (transporter affinity) but instead to a 45% to 75% increase in maximal transport rate (Vmax). The cytokine-stimulated increase in glutamine uptake by PAECs was completely blocked by actinomycin D and cycloheximide, indicating that the accelerated glutamine transport was dependent on de novo RNA and protein synthesis, perhaps of the transporter itself. The data indicate that these cytokines accelerate glutamine uptake by PAECs, either directly or indirectly, a response which may be required to support endothelial metabolism, structure, and function during infection and inflammation. The results of this study represent, to our knowledge, the first reports of cytokine-mediated modulation of System ASC activity, a carrier that has historically been unresponsive to hormonal regulation in other tissues.
Asunto(s)
Glutamina/metabolismo , Interleucina-1/farmacología , Arteria Pulmonar/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Animales , Transporte Biológico Activo/efectos de los fármacos , Células Cultivadas , Cicloheximida/farmacología , Dactinomicina/farmacología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Arteria Pulmonar/citología , Arteria Pulmonar/efectos de los fármacos , PorcinosRESUMEN
The alterations in lung glutamine (GLN) metabolism that occurs in the endotoxin-injured lung were studied in rats and subsequently correlated with flux changes that occur in patients with the adult respiratory distress syndrome (ARDS). Measurements in animals were made at various time-points following the administration of endotoxin, while studies in surgical patients were done in a group of healthy controls, in patients with "early" sepsis who had normal chest x-ray films, and in patients with radiographic and physiologic evidence of ARDS. In healthy control rats, net amounts of GLN are released by the lungs into the systemic circulation. This release rate doubled 30 minutes after intravenous endotoxin (1,580 +/- 320 nmol GLN/100 g BW/min vs. 736 +/- 179 in controls, p less than 0.01) but glutamine synthetase activity was unchanged, suggesting an outpouring of cellular glutamine stores. Two hours after endotoxin treatment, this accelerated fractional release of glutamine by the lungs was no longer detected. By the 12-hour time-point, the lungs reversed to an organ of net glutamine balance (234 +/- 248 nmol/100 g BW/min, p less than 0.05 vs. controls and ENDO30 min) despite a more than two-fold increase in glutamine synthetase activity (p less than 0.01). Simultaneously, lung weights were increased by 21% (p less than 0.01) and histologic examination showed an interstitial infiltrate and pulmonary edema. Similar observations were made in humans; patients with "early" sepsis exhibited a marked increase in lung glutamine release, while patients with ARDS demonstrated glutamine balance across the lungs (4,030 +/- 910 nmol GLN/kg BW/min vs. 637 +/- 496 in ARDS, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Endotoxinas/farmacología , Glutamina/metabolismo , Pulmón/metabolismo , Animales , Glutamato-Amoníaco Ligasa/metabolismo , Humanos , Infecciones/metabolismo , Pulmón/efectos de los fármacos , Pulmón/patología , Masculino , Ratas , Ratas Endogámicas , Síndrome de Dificultad Respiratoria/metabolismo , Procedimientos Quirúrgicos OperativosRESUMEN
We examined the alterations in brush-border glutamine transport that occurred in a surgically defunctionalized jejunal limb excluded from mucosal food contact. Dogs were surgically prepared with Roux-en-Y gastrojejunostomies to permit same-intestine comparisons of glutamine transport and glutaminase activity in jejunal segments that were in incontinuity or excluded for a 6-mo period. Transport of glutamine, alanine, and glucose was measured in brush-border membrane vesicles prepared from each intestinal section; membrane marker enzymes were enriched to the same degree in incontinuity and excluded portions. The Na(+)-dependent glutamine cotransport apparent Km was the same in the excluded (779 +/- 63 microM) and incontinuity (873 +/- 105 microM) limbs. However, the Jmax for Na(+)-independent glutamine transport in the incontinuity jejunum (158.7 +/- 15.7 pmol.mg protein-1.s-1) was double that in the excluded limb (71.2 +/- 4.6 pmol.mg protein-1.s-1). Na(+)-dependent carrier-mediated glutamine transport rates were lower than the Na(+)-dependent system, but Na(+)-independent kinetic parameters were not significantly different in incontinuity vs. excluded limbs (Jmax 7.9 +/- 0.6 pmol.mg protein-1.s-1; Km 140 +/- 20 microM). Similarly, the passive diffusion permeability coefficient was the same for both excluded and incontinuity jejunal limbs (22.7 +/- 0.9 nl.mg protein-1.s-1). Mucosal glutaminase enzyme activity was increased by 28% in the incontinuity limb (4.32 +/- 0.21 vs. 3.36 +/- 0.35 mumol.mg protein-1.h-1; P less than 0.02). Transport rates of alanine and glucose were also diminished in the excluded limb (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Glutamina/metabolismo , Yeyuno/metabolismo , Alanina/metabolismo , Anastomosis en-Y de Roux , Asparagina/metabolismo , Transporte Biológico Activo , Glucosa/metabolismo , Glutaminasa/metabolismo , Histidina/metabolismo , Yeyuno/cirugía , Yeyuno/ultraestructura , Cinética , Microvellosidades/enzimología , Microvellosidades/metabolismo , Sodio/fisiologíaRESUMEN
The effects of severe infection on luminal transport of amino acids and glucose by the small intestine were investigated. Studies were done in endotoxin-treated rats and in septic patients who underwent resection of otherwise normal small bowel. In rats the kinetics of the brush border glutamine transporter and the glutaminase enzyme were examined. In patients the effects of severe infection on the transport of glutamine, alanine, leucine, and glucose were studied. Transport was measured using small intestinal brush border membrane vesicles that were prepared by Mg++ aggregation/differential centrifugation. Uptake of radiolabeled substrate was measured using a rapid mixing/filtration technique. Vesicles demonstrated 15-fold enrichments of enzyme markers, classic overshoots, transport into an osmotically active space, and similar 2-hour equilibrium values. The sodium-dependent pathway accounted for nearly 90% of total carrier-mediated transport. Kinetic studies on rat jejunal glutaminase indicated a decrease in activity as early as 2 hours after endotoxin secondary to a decrease in enzyme affinity for glutamine (Km = 2.23 +/- 0.20 mmol/L [millimolar] in controls versus 4.55 +/- 0.67 in endotoxin, p less than 0.03), rather than a change in Vmax. By 12 hours the decrease in glutaminase activity was due to a decrease in Vmax (222 +/- 36 nmol/mg protein/min in controls versus 96 +/- 16 in endotoxin, p less than 0.03) rather than a significant change in Km. Transport data indicated a decrease in sodium-dependent jejunal glutamine uptake 12 hours after endotoxin secondary to a 35% reduction in maximal transport velocity (Vmax = 325 +/- 12 pmol/mg protein/10 sec in controls versus 214 +/- 8 in endotoxin, p less than 0.0001) with no change in Km (carrier affinity). Sodium-dependent glutamine transport was also decreased in septic patients, both in the jejunum (Vmax for control jejunum = 786 +/- 96 pmol/mg protein/10 sec versus 417 +/- 43 for septic jejunum, p less than 0.01) and in the ileum (Vmax of control ileum = 1126 +/- 66 pmol/mg protein/10 sec versus 415 +/- 24 in septic ileum, p less than 0.001) The rate of jejunal transport of alanine, leucine, and glucose was also decreased in septic patients by 30% to 50% (p less than 0.01). These data suggest that there is a generalized down-regulation of sodium-dependent carrier-mediated substrate transport across the brush border during severe infection, which probably occurs secondary to a decrease in transporter synthesis or an increase in the rate of carrier degradation.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Infecciones por Escherichia coli/metabolismo , Glutamina/metabolismo , Intestino Delgado/cirugía , Microvellosidades/metabolismo , Sepsis/metabolismo , Adulto , Animales , Transporte Biológico , Endotoxinas/toxicidad , Glutaminasa/metabolismo , Humanos , Intestino Delgado/enzimología , Cinética , Masculino , Ratas , Ratas Endogámicas , Sepsis/inducido químicamenteRESUMEN
Intestinal extraction of circulating glutamine across the basolateral membrane is diminished in the tumor-bearing rat (TBR). This study was designed to investigate the effects of progressive malignant growth on brush border glutamine transport in order to gain further insight into the adaptive/regulatory changes in intestinal glutamine metabolism that occur in the tumor-bearing rat. Fischer 344 rats (225 +/- 5 g) were implanted with fibrosarcoma cells and were studied at various time points after implantation when the tumors comprised 7%, 20%, and 29% of total body weight. Control and tumor-bearing rats were pair-fed throughout the study. Jejunal brush border membrane vesicles (BBMVs) were prepared by magnesium aggregation/differential centrifugation and transport of radioactively labeled L-glutamine, L-leucine, L-alanine, and D-glucose by BBMVs was measured using a Millipore filtration technique. BBMVs were enriched 15-fold in alkaline phosphatase, indicating brush border vesicle purity. Uptake of all substrates occurred into an osmotically active space, exhibited overshoots, and had similar 1-hr equilibrium values. The rate of glutamine uptake by BBMVs from all tumor-bearing rats was significantly greater than controls, regardless of tumor size. The increase in transport activity was not due to a change in carrier affinity but rather to an increase in maximal transport velocity. In rats with small tumors (7% of body weight), the Vmax was 431 +/- 40 pmole/mg protein/10 sec compared to 259 +/- 30 in control animals (P less than 0.01). In marked contrast, the mean transport of alanine was diminished in BBMVs from TBR (31 +/- 3 pmole/mg protein/10 sec in TBR vs 23 +/- 2 in controls, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Glutamina/metabolismo , Absorción Intestinal , Microvellosidades/metabolismo , Neoplasias Experimentales/metabolismo , Animales , Transporte Biológico , Proteínas Portadoras/metabolismo , Técnicas In Vitro , Cinética , Neoplasias Experimentales/patología , Ratas , Ratas Endogámicas F344RESUMEN
Fast-growing tumors are major glutamine consumers and may alter host glutamine metabolism to benefit the tumor. Previous studies from our laboratory have demonstrated that the liver switches from an organ of glutamine balance to one of glutamine release with progressive malignant growth. However, the regulation of this change is unclear. This study examined tumor modulation of hepatic glutamine metabolism by determining the activities of glutaminase, the principle enzyme of glutamine degradation, and glutamine synthetase, the principal enzyme of glutamine synthesis. Hepatic glutamine content was also determined. Rats with a fast-growing subcutaneous fibrosarcoma (TBR) and pair-fed controls were studied at 2 and 3 weeks after tumor or sham implantation, when the tumors comprised approximately 5% and 20% of total body weight. Arterial glutamine fell with progressive tumor growth (608 +/- 26 mumol/L in controls vs 494 +/- 15 in TBR, p less than 0.005) and was not attributable to a diminished food intake. Hepatic glutamine content was increased 45% (p less than 0.01) in tumor rats at 2 weeks due in part to a 35% fall in liver glutaminase activity. At 3 weeks, glutamine synthetase activity increased by 43% (0.58 +/- 0.07 mumol/mg of protein/hr in controls vs 0.83 +/- 0.04 in TBR, p less than 0.01) whereas glutaminase remained depressed (2.68 +/- 0.12 mumol/mg of protein/hr in controls vs 2.22 +/- 0.15 in TBR, p less than 0.05) and glutamine content fell compared to 2 week tumor-bearing rats, consistent with accelerated hepatic glutamine release. Tumors may alter liver glutamine metabolism by modulating hepatic enzyme activity in order to provide circulating glutamine for the growing malignancy.