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BACKGROUND: Citral (C10H16O) is the main ingredient of Cymbopogon citratus (lemongrass oil) and can reduce the side effects of oxidative stress. Diabetes caused by insulin deficiency induces oxidative stress in the liver. AIMS: This study aimed to investigate the ameliorative effects of citral on selected oxidative parameters and the gene expression of paraoxonase 1 (PON1) and endothelial nitric oxide synthase (eNOS) in a rat model of streptozotocin (STZ)-induced diabetes mellitus. METHODS: Forty rats were divided into four groups at random: control (C), control citral (CC), and two STZ-induced diabetic groups (diabetic (D) and citral diabetic (CD)). After diabetes confirmation (day 7), gavage treatment with citral (300 mg/kg body weight (BW)) was started in the CD and CC groups and continued for two weeks. RESULTS: On day 21 of the study, following treatment with citral for 14 days, the serum levels of total antioxidant capacity (TAC), and PON1 in the CD group were significantly increased compared to those in the D group (P<0.05). While treatment with citral caused a significant decrease in the Malondialdehyde (MDA), and eNOS in the CD group compared to those of the D group (P<0.001). The expression rate of liver PON1 gene was considerably upregulated in the CD group compared to that in the D group (P<0.001); while the opposite was observed for eNOS gene expression. However, there was no significant difference between the CC and C groups in terms of all examined parameters (P>0.05). CONCLUSION: This study showed positive effects of citral on serum antioxidant status and liver gene expression of PON1 and eNOS in diabetic rats.
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BACKGROUND: The higher incidence of obstructive sleep apnea (OSA) in men than in women has been attributed to the upper airway being longer in men. The Starling resistor is the paradigm biomechanical model of upper airway collapse in OSA where a collapsible tube (representing the pharynx) is located between two rigid tubes (representing the nasal cavity and trachea). While the Starling resistor has been extensively studied due to its relevance to many physiological phenomena, the effect of tube length on tube collapsibility has not been quantified yet. METHODS: Finite element analysis of a 3-dimensional collapsible tube subjected to a transmural pressure was performed in ANSYS Workbench. The numerical methods were validated with in vitro experiments in a silicone tube whose modulus of elasticity (361 ± 28 kPa) and dimensions (length = 100 mm, diameter = 22.2 mm, and wall thickness = 1.59 mm) were selected so that tube compliance was similar to pharyngeal compliance in humans during sleep. The buckling pressure (transmural pressure at which the tube collapses) was quantified in tubes of three different diameters (10 mm, 16 mm, and 22.2 mm) and ten length-to-diameter ratios (L/D = 4 to 13), while keeping the wall-thickness-to-radius ratio constant at 0.143. RESULTS: The absolute value of the buckling pressure decreased from 4.7 to 3.3 cmH2O (461-324 Pa) when L/D increased from 4 to 13. The buckling pressure was nearly independent from tube length for L/D >10. CONCLUSIONS: Our finding that longer tubes are more collapsible than shorter tubes is consistent with the higher incidence of obstructive sleep apnea in males than females.
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Faringe , Apnea Obstructiva del Sueño , Elasticidad , Femenino , Humanos , Masculino , Presión , SueñoRESUMEN
The potentially pathogenic Non-Tuberculosis Mycobacteria (NTM) are emerging nowadays which result in pulmonary and non-pulmonary infections in human. This group of bacteria consists of at least 200 different species. While the pulmonary disease is the most common form of NTM infections, NTM can cause diffused infections as well as extrapulmonary infections in every organ, such as bone marrow, skin, eye, and brain. The NTM cause tuberculosis-like infections, therefore, correct identification of these Mycobacteria is necessary to avoid faulty treatment. Different species of NTM isolates were identified from clinical specimens using phenotypic methods and Line Probe Assay. Minimum Inhibitory Concentration for selected antibiotics was obtained by the broth micro-dilution method. Totally, 42 NTM isolates were identified in this study. Moreover, the frequency of NTM between all positive mycobacterium cultures was estimated at 12%. The most common Rapidly Growing Mycobacteria included Mycolicibacterium fortuitum (30.9%), Mycobacterium abscessus (7.1%), and Mycobacterium chelonae (2.3%), whereas Mycobacterium simiae (40.4%), Mycobacterium kansasii (16.6%), and Mycobacterium avium complex (2.3%) were the most recurring among the Slowly Growing Mycobacteria. Amikacin, clarithromycin, and ciprofloxacin were the most effective antibiotics against isolated NTM. The NTM isolates are frequently being separated from Iranian patients, and are mostly resistant to the wide spectrum of antibiotics. Correct identification and determination of antibiotic susceptibility can be helpful in the healing process of the patients who suffer from non-tuberculosis mycobacterial infections.
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Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Infecciones por Mycobacterium no Tuberculosas/epidemiología , Micobacterias no Tuberculosas/aislamiento & purificación , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Irán/epidemiología , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Infecciones por Mycobacterium no Tuberculosas/microbiología , Micobacterias no Tuberculosas/efectos de los fármacos , Micobacterias no Tuberculosas/fisiología , Prevalencia , Adulto JovenRESUMEN
Meniscal tear is one of the prevalent knee disorders among young athletes and the aging population, and requires correct diagnosis and surgical intervention, if necessary. Not only the errors followed by human intervention but also the obstacles of manual meniscal tear detection highlight the need for automatic detection techniques. This paper presents a type-2 fuzzy expert system for meniscal tear diagnosis using PD magnetic resonance images (MRI). The scheme of the proposed type-2 fuzzy image processing model is composed of three distinct modules: Pre-processing, Segmentation, and Classification. λ-nhancement algorithm is used to perform the pre-processing step. For the segmentation step, first, Interval Type-2 Fuzzy C-Means (IT2FCM) is applied to the images, outputs of which are then employed by Interval Type-2 Possibilistic C-Means (IT2PCM) to perform post-processes. Second stage concludes with re-estimation of "η" value to enhance IT2PCM. Finally, a Perceptron neural network with two hidden layers is used for Classification stage. The results of the proposed type-2 expert system have been compared with a well-known segmentation algorithm, approving the superiority of the proposed system in meniscal tear recognition.
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Algoritmos , Diagnóstico por Computador , Meniscos Tibiales/diagnóstico por imagen , Lesiones de Menisco Tibial/diagnóstico por imagen , Humanos , Procesamiento de Imagen Asistido por Computador , Imagen por Resonancia MagnéticaRESUMEN
The focus of this paper is diagnosing and differentiating Astrocytomas in MRI scans by developing an interval Type-2 fuzzy automated tumor detection system. This system consists of three modules: working memory, knowledge base, and inference engine. An image processing method with three steps of preprocessing, segmentation and feature extraction, and approximate reasoning is used in inference engine module to enhance the quality of MRI scans, segment them into desired regions, extract the required features, and finally diagnose and differentiate Astrocytomas. However, brain tumors have different characteristics in different planes, so considering one plane of patient's MRI scan may cause inaccurate results. Therefore, in the developed system, several consecutive planes are processed. The performance of this system is evaluated using 95 MRI scans and the results show good improvement in diagnosing and differentiating Astrocytomas.
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Astrocitoma/diagnóstico , Neoplasias Encefálicas/diagnóstico , Sistemas Especialistas/instrumentación , Procesamiento de Imagen Asistido por Computador/instrumentación , Imagen por Resonancia Magnética/métodos , Algoritmos , Astrocitoma/patología , Neoplasias Encefálicas/patología , Lógica Difusa , HumanosRESUMEN
In preclinical studies, antagonists of growth hormone-releasing hormone (GHRH) have demonstrated inhibitory effects on the growth of various types of cancers expressing the pituitary type of GHRH receptors (pGHRH-R) and/or its active splice variant 1 (SV1). In this study, we investigated the effectiveness of the treatment of MDA-MB-231 human triple-negative breast cancer (TNBC) with GHRH antagonist JMR-132 alone or in combination with docetaxel. Receptor expression in the MDA-MB-231 human breast cancer cell line was evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Cell viability assays were performed on MDA-MB-231 cells treated with JMR-132, docetaxel or in combination. For studies in vivo, a subcutaneous nude mouse xenograft model was used. JMR-132 was administered s.c. at a dose of 10 µg/day and docetaxel at a dose of 10 mg/kg i.p. given on day 1 and 5. Similar regimens were used for the combination of both substances. At the end of the experiment, an mRNA-based human cancer pathway array including 84 major genes was performed on the tumor tissue of mice treated with JMR-132 to elucidate the mechanism of action of GHRH antagonists in vivo. The in vitro proliferation studies revealed that JMR-132 and docetaxel decreased the cell viability in a dose-dependent manner. The combination of both treatments produced a significantly greater inhibition of cell viability compared to the single agents. Treatment of nude mice bearing MDA-MB-231 xenografts with JMR-132 and docetaxel significantly (p<0.05) inhibited tumor growth by 46 and 50%, respectively. Treatment with the combination of JMR-132 and docetaxel led to an inhibition of tumor volume by 71.6% (p<0.001). Polymerase chain reaction array analysis revealed that JMR-132 interacts with signal transduction pathways involved in proliferation, apoptosis and angiogenesis. Our results suggest that GHRH antagonists in combination with taxanes may enhance the efficacy of treatment for patients with TNBC expressing the SV1 and/or the pGHRH receptor.
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Antineoplásicos/uso terapéutico , Hormona Liberadora de Hormona del Crecimiento/antagonistas & inhibidores , Sermorelina/análogos & derivados , Taxoides/uso terapéutico , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Animales , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular/efectos de los fármacos , Docetaxel , Femenino , Humanos , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Neovascularización Patológica/tratamiento farmacológico , Receptores de Neuropéptido/metabolismo , Receptores de Hormona Reguladora de Hormona Hipofisaria/metabolismo , Sermorelina/uso terapéutico , Neoplasias de la Mama Triple Negativas/genética , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Epithelial ovarian carcinoma is the leading cause of cancer-related deaths among women with gynecologic malignancies. Antagonists of the growth hormone-releasing hormone (GHRH) have been shown to inhibit growth of various cancers through endocrine, autocrine, and paracrine mechanisms. In this study, we have investigated the effects of GHRH antagonists (GHRHa) in ES-2 human clear cell ovarian cancer and in UCI-107 human serous ovarian cancer in vitro and in vivo. We evaluated the expression of mRNA for GHRH receptor, the binding to GHRH receptors, in specimens of ES-2 ovarian cancer. We evaluated also the in vitro effects of GHRHa on ES-2 cells and the in vivo effect of 2 different GHRHa on ES-2 and UCI-107 tumors. Nude mice bearing xenografts on ES-2 and UCI-107 ovarian cancer were treated with JMR-132 and MZ-J-7-118, respectively. Tumor growth was compared to control. ES-2 cells expressed mRNA for the functional splice variant SV1 of the GHRH receptor. JMR-132 inhibited cell proliferation in vitro by 42% and 18% at 10 and 1 µM concentration, respectively. Specific high affinity receptors for GHRH were detected in ES-2 cancer samples. In vivo daily subcutaneous injections of GHRHa significantly reduced tumor growth compared to a control group in both animal models. Our results indicate that GHRHa such as JMR-132 and MZ-J-7-118 can inhibit the growth of human ovarian cancer. The efficacy of GHRHa in ovarian cancer should be assessed in clinical trials.
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Antineoplásicos/uso terapéutico , Hormona Liberadora de Hormona del Crecimiento/antagonistas & inhibidores , Antagonistas de Hormonas/uso terapéutico , Neoplasias Ováricas/tratamiento farmacológico , Animales , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Hormona Liberadora de Hormona del Crecimiento/genética , Hormona Liberadora de Hormona del Crecimiento/metabolismo , Antagonistas de Hormonas/metabolismo , Antagonistas de Hormonas/farmacología , Humanos , Ratones , Ratones Desnudos , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , ARN Mensajero/metabolismo , Distribución Aleatoria , Sermorelina/análogos & derivados , Sermorelina/farmacología , Sermorelina/uso terapéutico , Carga Tumoral/efectos de los fármacos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Short fragments and fragment analogues of beta-amyloid 1-42 peptide (Abeta1-42) display a protective effect against Abeta-mediated neurotoxicity. After consideration of our earlier results with in vitro bioassay of synthetic Abeta-recognition peptides and toxic fibrillar amyloids, five pentapeptides were selected as putative neuroprotective agents: Phe-Arg-His-Asp-Ser amide (Abeta4-8) and Gly-Arg-His-Asp-Ser amide (an analogue of Abeta4-8), Leu-Pro-Tyr-Phe-Asp amide (an analogue of Abeta17-21), Arg-Ile-Ile-Gly-Leu amide (an analogue of Abeta30-34), and Arg-Val-Val-Ile-Ala amide (an analogue of Abeta38-42). In vitro electrophysiological experiments on rat brain slices demonstrated that four of these peptides counteracted with the field excitatory postsynaptic potential-attenuating effect of Abeta1-42; only Arg-Val-Val-Ile-Ala amide proved inactive. In in vivo experiments using extracellular single-unit recordings combined with iontophoresis, all these pentapeptides except Arg-Val-Val-Ile-Ala amide protected neurons from the NMDA response-enhancing effect of Abeta1-42 in the hippocampal CA1 region. These results suggest that Abeta recognition sequences may serve as leads for the design of novel neuroprotective compounds.
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Péptidos beta-Amiloides/fisiología , Amiloide/fisiología , Neuronas/fisiología , Fármacos Neuroprotectores/farmacología , Oligopéptidos/fisiología , Fragmentos de Péptidos/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Péptidos beta-Amiloides/ultraestructura , Animales , Electrofisiología , Masculino , N-Metilaspartato/farmacología , Neuronas/efectos de los fármacos , Neuronas/ultraestructura , Fármacos Neuroprotectores/aislamiento & purificación , Fármacos Neuroprotectores/metabolismo , Oligopéptidos/aislamiento & purificación , Fragmentos de Péptidos/ultraestructura , Ratas , Ratas WistarRESUMEN
PURPOSE: We have postulated that the peptide domain(s) of the heparin-binding cytokine(s) might have biological activity, which theoretically could be exploited for modulation of the biological behavior of cancer cells. MATERIALS AND METHODS: We used HGF as a model heparin-binding cytokine and synthesized two HGF beta-chain domains, HHRGK (HGP1) and RYRNKH (HGP2), as well as four variants. As target cells, we used three cancer cell lines (HT25 human colonic, HT168-M1/9 human melanoma and 3LL-HH murine lung carcinoma) all characterized by strong liver metastatic potentials. The effects of peptides on cell proliferation, tumor growth and liver metastasis were evaluated. RESULTS: All the basic penta- or hexapeptides exhibited similar antiproliferative effects in vitro in a dose range of 100-1000 ng/ml. Meanwhile, none of the HGP peptides exhibited significant antitumoral effects on the primary spleen tumors in the form of systemic treatment. However, systemic treatment with HGP1, but not with HGP2, applied at the early phase of the dissemination process, showed an inhibitory effect on liver metastatization of all the tumor lines studied. Furthermore, one out of the four hexapeptides, BP4 (KRKRKR), had similar activity. CONCLUSION: Recent data on the antiangiogenic effects of these basic peptides partially explain the in vivo antimetastatic activity. We suggest the small basic penta-hexapeptides as a new class of biological response modifiers which can modulate the metastatic process.
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Adenocarcinoma/prevención & control , Adenocarcinoma/secundario , Factor de Crecimiento de Hepatocito/farmacología , Neoplasias Hepáticas/prevención & control , Neoplasias Hepáticas/secundario , Oligopéptidos/farmacología , Fragmentos de Péptidos/farmacología , Animales , Modelos Animales de Enfermedad , Humanos , Neoplasias Hepáticas Experimentales/prevención & control , Neoplasias Hepáticas Experimentales/secundario , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/patología , Melanoma/prevención & control , Melanoma/secundario , Ratones , Ratones Endogámicos C57BL , Ratones SCID , Estructura Terciaria de Proteína , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Fluorescent cell analytical techniques require the incorporation of a fluorophore into the target molecule without causing a significant change in the native conformation. Many short peptides have a limited number of reactive groups that can be labeled without affecting the biological activity. In this work we present several methods for labeling beta-amyloid peptides (betaA[25-35], betaA[1-40]) and their derivatives (LPFFD, RIIGL and RVVIA) with different chromophores exclusively at the N-terminus. In the case of liquid-phase labeling, fluorescein isothiocyanate was used. The side-chain amino function of Lys, if present in the sequence, was protected with an Fmoc group, whereby the hydrophobic character of the peptide was further increased. The labeling reaction was carried out in an appropriate deaggregating solvent, DMSO. For solid-phase labeling, 5(6)-carboxyfluorescein and 7-amino-4-methyl-3-coumarinylacetic acid were applied. Several cleavage cocktails were tested for removal of the labeled amyloid peptides from the resin in order to completely suppress the oxidation of Met.
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Péptidos beta-Amiloides/síntesis química , Bioquímica/métodos , Colorantes Fluorescentes/farmacología , Fragmentos de Péptidos/síntesis química , Acetonitrilos/química , Dimetilsulfóxido/farmacología , Fluoresceína-5-Isotiocianato/farmacología , Fluoresceínas/farmacología , Lisina/química , Modelos Químicos , Biosíntesis de Péptidos , Péptidos/química , Factores de TiempoRESUMEN
Ample experimental evidence suggests that beta-amyloid (A beta), when injected into the rat magnocellular nucleus basalis (MBN), impels excitotoxic injury of cholinergic projection neurons. Whereas learning and memory dysfunction is a hallmark of A beta-induced cholinergic deficits, anxiety, or hypoactivity under novel conditions cannot be attributed to the loss of cholinergic MBN neurons. As mood-related behavioral parameters are primarily influenced by the central serotonergic system, in the present study we investigated whether A beta(1-42) toxicity in the rat MBN leads to an altered serotonergic innervation pattern in the rat basal forebrain and cerebral cortex 7 days postsurgery. A beta infusion into the MBN elicited significant anxiety in the elevated plus maze. A beta toxicity on cholinergic MBN neurons, expressed as the loss of acetylcholinesterase-positive cortical projections, was accompanied by sprouting of serotonergic projection fibers in the MBN. In contrast, the loss of serotonin-positive fiber projections, decreased concentrations of both serotonin and 5-hydroxyindoleacetic acid, and decline of cortical 5-HT(1A) receptor binding sites indicated reduced serotonergic activity in the somatosensory cortex. In conclusion, the A beta-induced primary cholinergic deficit in the MBN and subsequent cortical cholinergic denervation bidirectionally modulate serotonergic parameters in the rat basal forebrain and cerebral cortex. We assume that enhanced serotonin immunoreactivity in the damaged MBN indicates intrinsic processes facilitating neuronal recovery and cellular repair mechanisms, while diminished cortical serotonergic activity correlates with the loss of the subcortical cholinergic input, thereby maintaining the balance of neurotransmitter concentrations in the cerebral cortex.
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Péptidos beta-Amiloides/farmacología , Núcleo Basal de Meynert/patología , Fibras Colinérgicas/patología , Fragmentos de Péptidos/farmacología , Serotonina/metabolismo , Corteza Somatosensorial/patología , 8-Hidroxi-2-(di-n-propilamino)tetralin/metabolismo , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Animales , Ansiedad/patología , Fibras Colinérgicas/química , Fibras Colinérgicas/metabolismo , Densitometría , Ácido Hidroxiindolacético/metabolismo , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Regeneración Nerviosa/fisiología , Vías Nerviosas/patología , Ensayo de Unión Radioligante , Ratas , Ratas Wistar , Receptores de Serotonina/análisis , Receptores de Serotonina/metabolismo , Receptores de Serotonina 5-HT1 , Agonistas de Receptores de Serotonina/metabolismo , Agonistas de Receptores de Serotonina/farmacología , Corteza Somatosensorial/metabolismo , TritioRESUMEN
One of the characteristic neuropathological hallmarks of Alzheimer's disease (AD) is the extracellular accumulation of beta-amyloid peptides (Abeta) in neuritic plaques. Experimental data indicate that different molecular forms of Abeta affect a wide array of neuronal and glial functions and thereby may lead to neuronal death in the nervous system. Whereas the fatal outcome of Abeta overproduction in transgenic cell lines, and of exogenous Abeta administration in numerous neurotoxicity models, is well established, particular facets of a complex molecular cascade by which Abeta attack neural cells are still elusive. In the present review we summarize recent knowledge on mechanisms of Abeta aggregation, its role in Abeta neurotoxicity, and binding of Abeta peptides to putative neuronal and glial receptors. Additionally, an integrative view on the interactions of Ca2+ -mediated excitotoxicity and free radical-induced oxidative stress in Abeta toxicity is provided. Furthermore, we survey advances of pharmacological investigations attempting to prevent and antagonize Abeta toxicity, or to promote neuronal regeneration following Abeta-induced neurotoxic insults. We distinguish two major classes of therapeutic approaches: conventional pharmacotherapy that employs blockade of known receptors, signal transduction pathways, and re-uptake of neurotransmitters, and direct targeting of neurotoxic Abeta by means of beta-sheet breakers, functional anti-Abeta peptides, and antibodies. Although a clinically relevant neuroprotective strategy is not yet available, sequential combination of drug regimens may provide prospects for effective antagonism of late-life Abeta burden and subsequent development of dementia.
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Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/efectos de los fármacos , Péptidos beta-Amiloides/metabolismo , Neuronas/metabolismo , Nootrópicos/farmacología , Péptidos beta-Amiloides/antagonistas & inhibidores , Péptidos beta-Amiloides/toxicidad , Apoptosis , Canales de Calcio , Señalización del Calcio , Radicales Libres , Humanos , Neurotoxinas/metabolismo , Estrés Oxidativo , Placa Amiloide/efectos de los fármacosRESUMEN
Whereas a cardinal role for beta-amyloid protein (Abeta) has been postulated as a major trigger of neuronal injury in Alzheimer's disease, the pathogenic mechanism by which Abeta deranges nerve cells remains largely elusive. Here we report correlative in vitro and in vivo evidence that an excitotoxic cascade mediates Abeta neurotoxicity in the rat magnocellular nucleus basalis (MBN). In vitro application of Abeta to astrocytes elicits rapid depolarization of astroglial membranes with a concomitant inhibition of glutamate uptake. In vivo Abeta infusion by way of microdialysis in the MBN revealed peak extracellular concentrations of excitatory amino acid neurotransmitters within 20-30 min. Abeta-triggered extracellular elevation of excitatory amino acids coincided with a significantly enhanced intracellular accumulation of Ca2+ in the Abeta injection area, as was demonstrated by 45Ca2+ autoradiography. In consequence of these acute processes delayed cell death in the MBN and persistent loss of cholinergic fibre projections to the neocortex appear as early as 3 days following the Abeta-induced toxic insult. Such a sequence of Abeta toxicity was effectively antagonized by the N-methyl-D-aspartate (NMDA) receptor ligand dizocilpine maleate (MK-801). Moreover, Abeta toxicity in the MBN decreases with advancing age that may be associated with the age-related loss of NMDA receptor expression in rats. In summary, the present results indicate that Abeta compromises neurons of the rat MBN via an excitotoxic pathway including astroglial depolarization, extracellular glutamate accumulation, NMDA receptor activation and an intracellular Ca2+ overload leading to cell death.
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Péptidos beta-Amiloides/toxicidad , Astrocitos/metabolismo , Núcleo Basal de Meynert/metabolismo , Ácido Glutámico/metabolismo , Enfermedad de Alzheimer/inducido químicamente , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/patología , Animales , Ácido Aspártico/metabolismo , Astrocitos/citología , Núcleo Basal de Meynert/patología , Calcio/metabolismo , Radioisótopos de Calcio , Células Cultivadas , Modelos Animales de Enfermedad , Maleato de Dizocilpina/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Microdiálisis , Degeneración Nerviosa/metabolismo , Degeneración Nerviosa/patología , Fármacos Neuroprotectores/farmacología , Neurotoxinas/toxicidad , Ratas , Receptores de N-Metil-D-Aspartato/metabolismo , Taurina/metabolismoRESUMEN
Insulin-like growth factors (IGFs) I and II are implicated in progression of various tumours including colorectal carcinomas. To interfere with the production of IGFs, we treated male nude mice bearing xenografts of HT-29 human colon cancer with various potent growth hormone-releasing hormone (GH-RH) antagonists. Twice daily injections of antagonist MZ-4-71, 10 microg intraperitoneally or 5 microg subcutaneously (s.c.) resulted in a significant 43-45% inhibition of tumour growth. Longer acting GH-RH antagonists, MZ-5-156 and JV-1-36 given once daily at doses of 20 microg s.c. produced a 43-58% decrease in volume and weight of cancers. Histological analyses of HT-29 cancers demonstrated that both a decreased cell proliferation and an increased apoptosis contributed to tumour inhibition. GH-RH antagonists did not change serum IGF-I or IGF-II levels, but significantly decreased IGF-II concentration and reduced mRNA expression for IGF-II in tumours. In vitro studies showed that HT-29 cells produced and secreted IGF-II into the medium, and addition of MZ-5-156 dose-dependently decreased IGF-II production by about 40% as well as proliferation of HT-29 cells. Our studies demonstrate that GH-RH antagonists inhibit growth of HT-29 human colon cancers in vivo and in vitro. The effect of GH-RH antagonists may be mediated through a reduced production and secretion of IGF-II by cancer cells.
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Neoplasias del Colon/tratamiento farmacológico , Hormona Liberadora de Hormona del Crecimiento/antagonistas & inhibidores , Factor II del Crecimiento Similar a la Insulina/biosíntesis , Proteínas de Neoplasias/biosíntesis , Animales , División Celular/efectos de los fármacos , Neoplasias del Colon/química , Neoplasias del Colon/patología , Hormona Liberadora de Hormona del Crecimiento/análogos & derivados , Hormona Liberadora de Hormona del Crecimiento/farmacología , Células HT29/efectos de los fármacos , Células HT29/metabolismo , Humanos , Factor I del Crecimiento Similar a la Insulina/análisis , Factor II del Crecimiento Similar a la Insulina/análisis , Masculino , Ratones , Ratones Desnudos , Región Organizadora del Nucléolo , ARN Mensajero/análisis , Sermorelina/análogos & derivados , Sermorelina/farmacología , Trasplante HeterólogoRESUMEN
The aggregation of beta-amyloid peptides is very important for their neurotoxic effect; standardization of the aggregation grade is necessary for biological experiments. Measurement of aggregation with physicochemical methods is a difficult task. The present work revealed that FT-IR can be used for studying the aggregation properties of beta-amyloid peptides and the effects of environmental variables (solvent, pH, ions, and temperature) on aggregation. In dimethyl sulfoxide or hexafluoroisopropanol, amyloid peptides are in a monomeric state; on dilution with phosphate buffer just before measurement is made, aggregation begins. A detailed two-dimensional FT-IR correlation spectroscopic study was made of the conformational transitions that occur during the aggregation of beta-amyloid peptides. Two processes (random/helix-to-beta-sheet and aggregation of beta-sheets) and multiple conformational states were observed before the most stable form was attained. beta-Amyloid peptides undergo decomposition in basic buffers containing Ca(2+); this process should be avoided during aging experiments.
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Péptidos beta-Amiloides/química , Péptidos beta-Amiloides/normas , Estabilidad de Medicamentos , Humanos , Concentración de Iones de Hidrógeno , Técnicas In Vitro , Sustancias Macromoleculares , Conformación Proteica , Estructura Secundaria de Proteína , Estándares de Referencia , Espectroscopía Infrarroja por Transformada de Fourier , TemperaturaRESUMEN
Although data exist that address the attempt to correlate noninvasive Doppler-derived pressure gradients with invasive catheter pressure gradients in patients with coarctation of the aorta, few data exist about stiffness of the proximal descending aorta (precoarctation) and its relation to these pressure measurements. In this study, an in vitro flow model of a simulated neonatal aorta with a coarctation was developed. Three proximal descending aortas of different stiffnesses were used. The stiffness index of the proximal descending aorta was calculated as beta = ln [systolic pressure/diastolic pressure/(systolic diameter - diastolic diameter)]. We evaluated pressure gradients obtained by continuous wave Doppler and standard catheter methods and looked at acceleration of flow velocity determined by pulsed wave Doppler in the 3 precoarctation segments of differing stiffnesses. Pressures in the proximal descending aorta (precoarctation) increased with increasing stiffness, ranging from 105 mm Hg (soft) to greater than 300 mm Hg (stiff). Continuous wave Doppler instantaneous pressure gradients overestimated the catheter instantaneous pressure gradients substantially (mean 41% +/- 19%). The stiffer the precoarctation segment, the more the degree of overestimation: soft, 0% to 63% (= 3.47); medium, 13% to 54% (beta = 4.42); and stiff, 43% to 66% (beta = 5.91). Inclusion of the precoarctation velocity [V1] component in the Bernoulli equation did not significantly improve the correlation or the agreement. An additional observation was that pullback catheter peak-to-peak gradients were higher than simultaneous peak-to-peak gradients. In the stiff aorta, this difference could be greater than 22 mm Hg (>19%). Acceleration of flow velocity toward the coarctation was evident by pulsed wave Doppler interrogation. Increasing the stiffness of the precoarctation segment also increased the degree of acceleration within this proximal segment: soft, 0.4 to 0.8 m/s; medium, 0.5 to 1. 4 m/s; and stiff, 0.7 to 1.5 m/s. These data suggest that increasing stiffness of the proximal descending aorta can alter the continuous wave detected Doppler gradient and although the gradient itself has increased, it may not predict accurately the true severity of the localized, most severely obstructed segment.
Asunto(s)
Aorta/fisiopatología , Coartación Aórtica/diagnóstico por imagen , Coartación Aórtica/fisiopatología , Ecocardiografía Doppler , Cateterismo Cardíaco , Humanos , Modelos Lineales , Modelos Cardiovasculares , Presión , Flujo Sanguíneo Regional , TransductoresRESUMEN
A putative tetrapeptide beta-amyloid (Abeta) antagonist (propionyl-Ile-Ile-Gly-Leu [Pr-IIGL]) based on the [31-34] sequence of Abeta was previously shown to rescue astrocytes from Abeta-induced membrane depolarization and subsequent long-term elevations of the intracellular Ca2+ concentration in vitro. Here we provide in vivo evidence that the Pr-IIGL tetrapeptide effectively attenuates the excitotoxic action of Abeta(1-42) on cholinergic neurons of the rat magnocellular nucleus basalis (MBN). We also demonstrate by means of microdialysis that administration of Pr-IIGL abolished Abeta(1-42)-induced increases in extracellular aspartate and glutamate concentrations in the MBN, which coincide with a significant preservation of cholinergic MBN neurons and their cortical projections. This neuroprotective effect was associated with preserved exploratory behavior in an open-field paradigm, and improved memory retention in a step-through passive avoidance task. Our data presented here indicate for the first time the efficacy of short, modified functional Abeta antagonists in ameliorating Abeta excitotoxicity in vivo.
Asunto(s)
Péptidos beta-Amiloides/toxicidad , Núcleo Basal de Meynert/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Oligopéptidos/farmacología , Fragmentos de Péptidos/toxicidad , Acetilcolinesterasa/metabolismo , Péptidos beta-Amiloides/antagonistas & inhibidores , Animales , Ácido Aspártico/metabolismo , Reacción de Prevención/efectos de los fármacos , Núcleo Basal de Meynert/anatomía & histología , Núcleo Basal de Meynert/metabolismo , Conducta Animal/efectos de los fármacos , Espacio Extracelular/efectos de los fármacos , Espacio Extracelular/metabolismo , Ácido Glutámico/metabolismo , Histocitoquímica , Masculino , Microdiálisis , Actividad Motora/efectos de los fármacos , Fragmentos de Péptidos/antagonistas & inhibidores , Ratas , Ratas Wistar , Taurina/metabolismoRESUMEN
Flow volumes in an in vitro model of the aorta with 3 different degrees of stiffness (stiff, moderately stiff, and compliant) proximal to a coarctation were calculated by using a digital color Doppler echocardiography flow calculation method that semiautomatically integrates spatial and temporal color flow velocity data. These flow volumes were compared with those obtained by the conventional pulsed Doppler method with reference to ultrasonic flowmeter. Flow volumes determined by the automated method agreed well with those obtained by ultrasonic flowmeter, even in this compliant aorta model with vessel size changing with pulsation, whereas the pulsed Doppler method overestimated the reference data, especially for more compliant descending aortic segments. The combination of flow data with continuous wave Doppler allows definition of effective orifice area for coarctation.
Asunto(s)
Coartación Aórtica/diagnóstico por imagen , Velocidad del Flujo Sanguíneo , Ecocardiografía Doppler en Color , Aorta , Procesamiento de Imagen Asistido por Computador , Técnicas In Vitro , Modelos Lineales , Modelos Cardiovasculares , Flujo PulsátilRESUMEN
Previous experimental data indicate the involvement of Ca(2+)-related excitotoxic processes, possibly mediated by N-Methyl-D-Aspartate (NMDA) receptors, in beta-amyloid (beta A) neurotoxicity. On the other hand, other lines of evidence support the view that free radical generation is a critical step in the beta A-induced neurodegenerative cascade. In the present study, therefore, a neuroprotective strategy was applied to explore the contributions of each of these pathways in beta A toxicity. beta A(1-42) was injected into the magnocellular nucleus basalis of rats, while neuroprotection was achieved by either single or combined administration of the NMDA receptor antagonist MK-801 (2.5 mg/kg) and/or a vitamin E and C complex (150 mg/kg). The degree of neurodegeneration was determined by testing the animals in consecutive series of behavioral tasks, including elevated plus maze, passive avoidance learning, small open-field and open-field paradigms, followed by acetylcholinesterase (AChE), choline-acetyltransferase (ChAT), and superoxide dismutase (SOD) biochemistry. beta A injected in the nucleus basalis elicited significant anxiety in the elevated plus maze, derangement of passive avoidance learning, and altered spontaneous behaviors in both open-field tasks. A significant decrease in both AChE and ChAT accompanied by a similar decrement of MnSOD, but not of Cu/ZnSOD provided neurochemical substrates for the behavioral changes. Each of the single drug administrations protected against the neurotoxic events, whereas the combined treatment failed to ameliorate beta A toxicity.
Asunto(s)
Péptidos beta-Amiloides/antagonistas & inhibidores , Maleato de Dizocilpina/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Depuradores de Radicales Libres/farmacología , Neuronas/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Sustancia Innominada/efectos de los fármacos , Acetilcolinesterasa/metabolismo , Péptidos beta-Amiloides/toxicidad , Animales , Ansiedad/patología , Ansiedad/fisiopatología , Reacción de Prevención/efectos de los fármacos , Conducta Animal/efectos de los fármacos , Colina O-Acetiltransferasa/metabolismo , Masculino , Microinyecciones , Neuronas/patología , Ratas , Ratas Wistar , Corteza Somatosensorial/enzimología , Sustancia Innominada/patología , Sustancia Innominada/fisiopatología , Superóxido Dismutasa/metabolismoRESUMEN
Some antagonists of human growth hormone-releasing hormone (hGH-RH) synthesized previously were shown to inhibit in vivo proliferation of various human cancers in nude mice. However, the activity of these analogs requires an increase to assure clinical efficacy. In an attempt to prepare hGH-RH antagonists with a high and protracted activity, we synthesized and biologically tested 22 antagonistic analogs of hGH-RH(1-29)NH2. The ability of the antagonists to inhibit hGH-RH-induced GH release was evaluated in vitro in a superfused rat pituitary system, as well as in vivo after i.v. injection into rats. The binding affinity of the peptides to GH-RH receptors also was determined. All antagonistic analogs had the common core sequence [PhAc-Tyr1,D-Arg2, Phe(4-Cl)6 (para-chlorophenylalanine), Abu15 (alpha-aminobutyric acid), Nle27]hGH-RH(1-29)NH2 and contained Arg, D-Arg, homoarginine (Har), norleucine (Nle), and other substitutions. The following analogs were determined to have a high and/or protracted antagonistic activity: [PhAc-Tyr1,D-Arg2,Phe(4-Cl)6,Arg9,Abu15,Nle27, D-Arg29]hGH-RH(1-29)NH2 (JV-1-10), [PhAc-Tyr1,D-Arg2,Phe(4-Cl)6, Abu15,Nle27,D-Arg28,Har29]hGH-RH(1-29)NH2 (MZ-6-55), [PhAc-Tyr1, D-Arg2,Phe(4-Cl)6,Arg9,Abu15,Nle27,D-Arg28,Har29 ]hGH-RH(1-29)NH2 (JV-1-36), and [PhAc-Tyr1,D-Arg2,Phe(4-Cl)6,Har9,Tyr(Me)10,Abu15, Nle27,D-Arg28,Har29]hGH-RH(1-29)NH2 (JV-1-38). Among the peptides tested, analog JV-1-36 showed the highest GH-RH antagonistic activity in vitro and also induced a strong and prolonged inhibition of GH release in vivo for at least 30 min. The antagonist JV-1-38 was slightly less potent than JV-1-36 both in vitro and in vivo but proved to be very long-acting in vivo, suppressing the GH-RH-induced GH release even after 60 min. High and protracted in vivo activities of these antagonists indicate an improvement over earlier GH-RH analogs. Some of these hGH-RH antagonists could find clinical applications in the treatment of cancers dependent on insulin-like growth factors I and II.