RESUMEN
The beneficial effect of compost and compost tea on plant growth and protection is mainly associated with the microbial diversity and the presence of bacteria with plant growth-promoting effect. PGPR are considered as eco-friendly bio-fertilizers that may reduce the use of chemical pesticides and fertilizers. Three composts (AT, A10, and A30) were previously prepared from industrial wastes (olive mill wastewater, olive pomace, coffee ground, and phosphogypsum). In the present study, we isolated three bacterial strains from the compost teas. The phylogenetic identification of these bacterial strains (B.AT, B.A10, and B.A30) showed that they correspond to Serratia liquefaciens (B.AT and B.A10) and Achromobacter spanius (B.A30) species. A further characterization of the PGPR traits of these bacteria showed that they produce siderophore, exopolysaccharides, and IAA. Their effect on potato plant growth, yields, and tuber quality was performed under field culture conditions. Results showed that these strains can be characterized as PGPR, the best effect on potato plant growth was observed with Serratia liquefaciens (B.AT), the best yield and tuber quality was observed with Serratia liquefaciens (B.A10) while bacterial treatment with Achromobacter spanius (B.A30) is a Cd-tolerant PGPR.
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Compostaje , Plaguicidas , Solanum tuberosum , Achromobacter , Bacterias , Bencenoacetamidas , Cadmio , Café , Fertilizantes , Residuos Industriales , Filogenia , Piperidonas , Sideróforos , Suelo/química , Microbiología del Suelo , Té , Aguas ResidualesRESUMEN
AIMS: Despite the advanced cancer treatments, there is increased resistance to chemotherapy and subsequent mortality. In lack of reliable data in monolayer cultures and animal models, researchers are shifting to 3D cancer spheroids, which represents the in vivo robust tumour morphology. Calcium is essential in cell signalling and proliferation. It is found that T-type calcium channels (TTCCs) are overexpressed in various cancer cells, supporting their increased proliferation. Many of the TTCCs blockers available could target other channels besides TTCCs, which can cause adverse effects. Therefore, we hypothesise that TTA-A2, a highly selective blocker towards TTCCs, can inhibit the growth of cancer spheroids, and provide an anti-cancer and an adjuvant role in cancer therapy. METHODS: We studied TTA-A2 and paclitaxel (PTX-control drug) in lung adenocarcinoma cell line- A549, cancer cells and human embryonic kidney cell line- HEK 293, control cell, in their monolayer and spheroids forms for viability, proliferation, morphology change, migration, and invasion-after 48-96 h of treatment. KEY FINDINGS: Though the results varied between the monolayer and spheroids studies, we found both anti-cancer as well as adjuvant effect of TTA-A2 in both the studies. TTA-A2 was able to inhibit the growth, viability, and metastasis of the cancer cells and spheroids. Differences in the results of two modes might explain that why drugs tested successfully in monolayer culture fail in clinical trials. SIGNIFICANCE: This study establishes the role of TTA-A2, a potent TTCC blocker as an anti-cancer and adjuvant drug in reducing the viability and metastasis of the cancer cells.
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Adenocarcinoma del Pulmón/patología , Antineoplásicos , Bencenoacetamidas/farmacología , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio Tipo T/efectos de los fármacos , Neoplasias Pulmonares/patología , Piridinas/farmacología , Células A549 , Adenocarcinoma del Pulmón/tratamiento farmacológico , Bencenoacetamidas/uso terapéutico , Canales de Calcio Tipo T/fisiología , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células HEK293 , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Invasividad Neoplásica/prevención & control , Piridinas/uso terapéuticoRESUMEN
BACKGROUND: Glutamine serves as an important nutrient with many cancer types displaying glutamine dependence. Following cellular uptake glutamine is converted to glutamate in a reaction catalysed by mitochondrial glutaminase. This glutamate has many uses, including acting as an anaplerotic substrate (via alpha-ketoglutarate) to replenish TCA cycle intermediates. CB-839 is a potent, selective, orally bioavailable inhibitor of glutaminase that has activity in Triple receptor-Negative Breast Cancer (TNBC) cell lines and evidence of efficacy in advanced TNBC patients. METHODS: A panel of eleven breast cancer cell lines was used to investigate the anti-proliferative effects of the glutaminase inhibitors CB-839 and BPTES in different types of culture medium, with or without additional pyruvate supplementation. The abundance of the TCA cycle intermediate fumarate was quantified as a measure if TCA cycle anaplerosis. Pyruvate secretion by TNBC cultures was then assessed with or without AZD3965, a monocarboxylate transporter 1 (MCT1) inhibitor. Finally, two dimensional (2D) monolayer and three dimensional (3D) spheroid assays were used to compare the effect of microenvironmental growth conditions on CB-839 activity. RESULTS: The anti-proliferative activity of CB-839 in a panel of breast cancer cell lines was similar to published reports, but with a major caveat; growth inhibition by CB-839 was strongly attenuated in culture medium containing pyruvate. This pyruvate-dependent attenuation was also observed with a related glutaminase inhibitor, BPTES. Studies demonstrated that exogenous pyruvate acted as an anaplerotic substrate preventing the decrease of fumarate in CB-839-treated conditions. Furthermore, endogenously produced pyruvate secreted by TNBC cell lines was able to act in a paracrine manner to significantly decrease the sensitivity of recipient cells to glutaminase inhibition. Suppression of pyruvate secretion using the MCT1 inhibitor AZD3965, antagonised this paracrine effect and increased CB-839 activity. Finally, CB-839 activity was significantly compromised in 3D compared with 2D TNBC culture models, suggesting that 3D microenvironmental features impair glutaminase inhibitor responsiveness. CONCLUSION: This study highlights the potential influence that both circulating and tumour-derived pyruvate can have on glutaminase inhibitor efficacy. Furthermore, it highlights the benefits of 3D spheroid cultures to model the features of the tumour microenvironment and improve the in vitro investigation of cancer metabolism-targeted therapeutics.
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Bencenoacetamidas/farmacología , Resistencia a Antineoplásicos , Glutaminasa/antagonistas & inhibidores , Glutamina/metabolismo , Ácido Pirúvico/metabolismo , Tiadiazoles/farmacología , Neoplasias de la Mama Triple Negativas/patología , Proliferación Celular , Femenino , Humanos , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Neoplasias de la Mama Triple Negativas/metabolismo , Células Tumorales Cultivadas , Microambiente TumoralRESUMEN
The aim of this study was to investigate the estrogen-like effects of 2-phenylacetamide (PA), which is the main compound isolated from the seeds of Lepidium apetalum Willd (LA). Results showed that LA and PA could promote the proliferation of MCF-7 cells. The mouse uterine weight test showed that, LA and PA could increase the uterus index of immature female mice, and the levels of luteinizing hormone (LH) and estrogen (E2). LA could increase the expression of ERα and ERß, while PA could increase the expression of ERα, ERß and GPR30 in the uterus and MCF-7 cells. In addition, co-incubation of the estrogen receptor blocker with LA or PA abolished the inductive effect of the proliferation. PA has estrogenic activities and was the material basis of LA that played the estrogenic effect. LA and PA might be used for the treatment of perimenopause syndrome in a novel application.
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Bencenoacetamidas/aislamiento & purificación , Bencenoacetamidas/farmacología , Estrógenos/farmacología , Lepidium/embriología , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Semillas/química , Animales , Proliferación Celular/efectos de los fármacos , Femenino , Humanos , Hormona Luteinizante/metabolismo , Células MCF-7 , Ratones , Receptores de Estrógenos/antagonistas & inhibidores , Receptores de Estrógenos/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Útero/efectos de los fármacos , Útero/metabolismoRESUMEN
PURPOSE: To evaluates analgesic effectiveness of 0.1% nepafenac during cataract surgery. METHODS: This prospective randomized randomized double-masked, placebo-controlled study comprised 80 eyes of 40 consecutive patients who underwent bilateral cataract surgery and implantation of foldable intraocular lens with topical anesthesia with and without topical nepafenac drops. Each eye of patients was assigned to group 1 and group 2. Topical anesthesia combined with 0.1% nepafenac used three times a day the day before the surgery and once half an hour just before the surgery was group 1, consisting of 40 eyes, and topical anesthesia with using placebo was group 2 consisting of 40 eyes. Patients were asked to score their pain using a visual analog scale (VAS) and verbal pain scale (VPS) immediately following the surgery. When the patient moved or squeeze the eye during surgery, the surgical comfort was evaluated as bad and otherwise, it was evaluated as good. RESULTS: When the intensity of pain during the surgery was evaluated, the percentage of patients reporting mild or no pain in group 1 was %825 and in group 2 was %45. Mean VAS pain score and mean VPS pain score in group 1 was significantly lower than that in group 2(pâ¯=â¯0.024, pâ¯<â¯0.001). Surgical comfort in group 1 was %825 and in group 2%65(Pâ¯=â¯0.075). CONCLUSION: 0.1% nepafenac reduces pain of patients who undergone routine clear corneal phacoemulsification with topical anesthesia and may increase patient comfort during the surgery when used preoperatively.
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Anestesia Local/métodos , Bencenoacetamidas/administración & dosificación , Extracción de Catarata/métodos , Dolor Postoperatorio/prevención & control , Satisfacción del Paciente , Fenilacetatos/administración & dosificación , Administración Tópica , Anciano , Antiinflamatorios no Esteroideos/administración & dosificación , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Estudios de Seguimiento , Humanos , Periodo Intraoperatorio , Masculino , Persona de Mediana Edad , Dimensión del Dolor , Estudios Prospectivos , Resultado del TratamientoRESUMEN
Chronic hyperglycemia increases apoptosis and reduces glucose-stimulated insulin secretion. Although protective agents have been searched extensively, none has been found so far. Here we tested FLZ, a synthetic derivative of squamosamide from a Chinese herb, as a potential candidate for antiglucotoxicity in INS-1E cells and mouse islets. Chronic culture of ß-cells in 30 mM glucose caused progressive reduction of cell viability, accompanied with increased apoptosis and reduced insulin secretion. These effects on apoptosis and insulin were reversed by FLZ in a dose-dependent manner. FLZ treatment also increased forkhead box O1 protein phosphorylation and reduced its nuclear location. On the contrary, FLZ increased pancreatic and duodenal homeobox-1 expression and its nuclear localization, an effect mediated by increased p-Akt. Consistently, Akt selective inhibitor MK-2206 completely abolished antiglucotoxicity effect of FLZ. Furthermore, FLZ treatment increased cytosolic ATP/ADP ratio. Taken together, our results suggest that FLZ could be a potential therapeutic agent to treat the hyperglycemia-induced ß-cell failure.
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Apoptosis/efectos de los fármacos , Bencenoacetamidas/farmacología , Medicamentos Herbarios Chinos/farmacología , Factores de Transcripción Forkhead/efectos de los fármacos , Glucosa/toxicidad , Células Secretoras de Insulina/efectos de los fármacos , Fenoles/farmacología , Proteínas Proto-Oncogénicas c-akt/efectos de los fármacos , Animales , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Proteína Forkhead Box O1 , Factores de Transcripción Forkhead/metabolismo , Insulina/metabolismo , Secreción de Insulina , Células Secretoras de Insulina/metabolismo , Islotes Pancreáticos/efectos de los fármacos , Islotes Pancreáticos/metabolismo , Ratones , Proteínas del Tejido Nervioso/efectos de los fármacos , Proteínas del Tejido Nervioso/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , RatasRESUMEN
Differential modulation of kappa opioid receptor (KOR) signaling has been a proposed strategy for developing therapies for drug addiction and depression by either activating or blocking this receptor. Hence, there have been significant efforts to generate ligands with diverse pharmacological properties including partial agonists, antagonists, allosteric modulators as well as ligands that selectively activate some pathways while not engaging others (biased agonists). It is becoming increasingly evident that G protein coupled receptor signaling events are context dependent and that what may occur in cell based assays may not be fully indicative of signaling events that occur in the naturally occurring environment. As new ligands are developed, it is important to assess their signaling capacity in relevant endogenous systems in comparison to the performance of endogenous agonists. Since KOR is considered the cognate receptor for dynorphin peptides we have evaluated the selectivity profiles of dynorphin peptides in wild-type (WT), KOR knockout (KOR-KO), and mu opioid receptor knockout (MOR-KO) mice using [35S]GTPγS binding assay in striatal membrane preparations. We find that while the small molecule KOR agonist U69,593, is very selective for KOR, dynorphin peptides promiscuously stimulate G protein signaling in striatum. Furthermore, our studies demonstrate that norBNI and 5'GNTI are highly nonselective antagonists as they maintain full potency and efficacy against dynorphin signaling in the absence of KOR. Characterization of a new KOR antagonist, which may be more selective than NorBNI and 5'GNTI, is presented using this approach.
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Cuerpo Estriado/efectos de los fármacos , Evaluación Preclínica de Medicamentos/métodos , Dinorfinas/metabolismo , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Receptores Opioides kappa/metabolismo , Analgésicos Opioides/farmacología , Animales , Bencenoacetamidas/farmacología , Cuerpo Estriado/metabolismo , Dinorfinas/administración & dosificación , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Antagonistas de Narcóticos/farmacología , Unión Proteica , Pirrolidinas/farmacología , Receptores Opioides kappa/agonistas , Receptores Opioides kappa/antagonistas & inhibidores , Receptores Opioides kappa/genética , Transducción de Señal/efectos de los fármacos , Radioisótopos de AzufreRESUMEN
The 5-lipoxygenase product 5-oxo-6,8,11,14-eicosatetraenoic acid (5-oxo-ETE) is the most powerful human eosinophil chemoattractant among lipid mediators and could play a major pathophysiological role in eosinophilic diseases such as asthma. Its actions are mediated by the OXE receptor, orthologs of which are found in many species from humans to fish, but not rodents. The unavailability of rodent models to examine the pathophysiological roles of 5-oxo-ETE and the OXE receptor has substantially hampered progress in this area. As an alternative, we have explored the possibility that the cat could serve as an appropriate animal model to investigate the role of 5-oxo-ETE. We found that feline peripheral blood leukocytes synthesize 5-oxo-ETE and that physiologically relevant levels of 5-oxo-ETE are present in bronchoalveolar lavage fluid from cats with experimentally induced asthma. 5-Oxo-ETE (EC50, 0.7nM) is a much more potent activator of actin polymerization in feline eosinophils than various other eicosanoids, including leukotriene (LT) B4 and prostaglandin D2. 5-Oxo-ETE and LTB4 induce feline leukocyte migration to similar extents at low concentrations (1nM), but at higher concentrations the response to 5-oxo-ETE is much greater. Although high concentrations of selective human OXE receptor antagonists blocked 5-oxo-ETE-induced actin polymerization in feline granulocytes, their potencies were about 200 times lower than for human granulocytes. We conclude that feline leukocytes synthesize and respond to 5-oxo-ETE, which could potentially play an important role in feline asthma, a common condition in this species. The cat could serve as a useful animal model to investigate the pathophysiological role of 5-oxo-ETE.
Asunto(s)
Ácidos Araquidónicos/farmacología , Asma/metabolismo , Eosinófilos/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Actinas/genética , Actinas/metabolismo , Alérgenos/inmunología , Animales , Araquidonato 5-Lipooxigenasa/genética , Araquidonato 5-Lipooxigenasa/metabolismo , Ácidos Araquidónicos/biosíntesis , Asma/inducido químicamente , Asma/genética , Asma/inmunología , Bencenoacetamidas/farmacología , Benzotiazoles/farmacología , Líquido del Lavado Bronquioalveolar/citología , Gatos , Quimiotaxis/efectos de los fármacos , Quimiotaxis/inmunología , Cynodon/química , Cynodon/inmunología , Modelos Animales de Enfermedad , Eosinófilos/metabolismo , Eosinófilos/patología , Femenino , Expresión Génica , Humanos , Leucotrieno B4/farmacología , Masculino , Neutrófilos/metabolismo , Neutrófilos/patología , Polimerizacion , Cultivo Primario de Células , Prostaglandina D2/farmacología , Receptores Eicosanoides/antagonistas & inhibidores , Receptores Eicosanoides/genética , Receptores Eicosanoides/metabolismoRESUMEN
BACKGROUND: Antineoplastons are naturally occurring peptides and amino acid derivatives found in human blood and urine. Antineoplaston A10 and AS2-1 reportedly control neoplastic growth and do not significantly inhibit normal cell growth. Antineoplastons contain 3-phenylacetylamino-2, 6-piperidinedione (A10), phenylacetylglutamine plus phenylacetylisoglutamine (A10-I), and phenylacetylglutamine plus phenylacetate (AS2-1). This open label, non- blinded randomized phase II study compared the efficacy of hepatic arterial infusion (HAI) with 5-fluorouracil,with or without antineoplastons as a postoperative therapy for colorectal metastasis to the liver. METHODS: Sixty-five patients with histologically confirmed metastatic colon adenocarcinoma in liver, who had undergone hepatectomy, and/or thermal ablation for liver metastases were enrolled between 1998- 2004 in Kurume University Hospital. Patients were randomly assigned to receive systemic antineoplastons (A10-I infusion followed by per-oral AS2-1) plus HAI (AN arm) or HAI alone (control arm) based on the number of metastases and presence/ absence of extra-hepatic metastasis at the time of surgery. Primary endpoint was cancer-specific survival (CSS); secondary endpoints were relapse-free survival (RFS), status and extent of recurrence, salvage surgery (rate) and toxicity. FINDINGS: Overall survival was not statistically improved (p=0.105) in the AN arm (n=32). RFS was not significant (p=0.343). Nevertheless, the CSS rate was significantly higher in the AN arm versus the control arm (n=33) with a median survival time 67 months (95%CI 43-not calculated) versus 39 months (95%CI 28-47) (p=0.037) and 5 year CSS rate 60% versus 32% respectively. Cancer recurred more often in a single organ than in multiple organs in the AN arm versus the control arm. The limited extent of recurrent tumours in the AN arm meant more patients remained eligible for salvage surgery. Major adverse effects of antineoplastons were fullness of the stomach and phlebitis. No serious toxicity, including bone marrow suppression, liver or renal dysfunction, were found in the AN arm. INTERPRETATION: Antineoplastons (A10 Injection and AS2-1) might be useful as adjunctive therapy in addition to HAI after hepatectomy in colorectal metastases to the liver. TRIAL REGISTRATION INFORMATION: ClinicalTrials.gov UMIN000012099.
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Antimetabolitos Antineoplásicos/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Neoplasias Colorrectales/tratamiento farmacológico , Fluorouracilo/administración & dosificación , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/secundario , Anciano , Antimetabolitos Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Bencenoacetamidas/administración & dosificación , Bencenoacetamidas/uso terapéutico , Quimioterapia Adyuvante/métodos , Neoplasias Colorrectales/cirugía , Combinación de Medicamentos , Fluorouracilo/uso terapéutico , Glutamina/administración & dosificación , Glutamina/análogos & derivados , Glutamina/uso terapéutico , Hepatectomía , Humanos , Infusiones Intraarteriales , Neoplasias Hepáticas/cirugía , Persona de Mediana Edad , Fenilacetatos/administración & dosificación , Fenilacetatos/uso terapéutico , Piperidonas/administración & dosificación , Piperidonas/uso terapéutico , Análisis de Supervivencia , Resultado del TratamientoRESUMEN
BACKGROUND AND PURPOSE: For patients experiencing inadequate analgesia and intolerable opioid-related side effects on one strong opioid analgesic, pain relief with acceptable tolerability is often achieved by rotation to a second strong opioid. These observations suggest subtle pharmacodynamic differences between opioids in vivo. This study in rats was designed to assess differences between opioids in their in vivo profiles. EXPERIMENTAL APPROACH: Male Sprague Dawley rats were given single i.c.v. bolus doses of morphine, morphine-6-glucuronide (M6G), fentanyl, oxycodone, buprenorphine, DPDPE ([D-penicillamine(2,5) ]-enkephalin) or U69,593. Antinociception, constipation and respiratory depression were assessed using the warm water tail-flick test, the castor oil-induced diarrhoea test and whole body plethysmography respectively. KEY RESULTS: These opioid agonists produced dose-dependent antinociception, constipation and respiratory depression. For antinociception, morphine, fentanyl and oxycodone were full agonists, buprenorphine and M6G were partial agonists, whereas DPDPE and U69,593 had low potency. For constipation, M6G, fentanyl and buprenorphine were full agonists, oxycodone was a partial agonist, morphine produced a bell-shaped dose-response curve, whereas DPDPE and U69,593 were inactive. For respiratory depression, morphine, M6G, fentanyl and buprenorphine were full agonists, oxycodone was a partial agonist, whereas DPDPE and U69,593 were inactive. The respiratory depressant effects of fentanyl and oxycodone were of short duration, whereas morphine, M6G and buprenorphine evoked prolonged respiratory depression. CONCLUSION AND IMPLICATIONS: For the seven opioids we assessed, no two had the same profile for evoking antinociception, constipation and respiratory depression, suggesting that these effects are differentially regulated. Our findings may explain the clinical success of 'opioid rotation'. LINKED ARTICLES: This article is part of a themed section on Opioids: New Pathways to Functional Selectivity. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2015.172.issue-2.
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Analgésicos Opioides/efectos adversos , Analgésicos Opioides/uso terapéutico , Estreñimiento/inducido químicamente , Dolor/tratamiento farmacológico , Insuficiencia Respiratoria/inducido químicamente , Animales , Bencenoacetamidas/efectos adversos , Bencenoacetamidas/uso terapéutico , Buprenorfina/efectos adversos , Buprenorfina/uso terapéutico , Aceite de Ricino , Diarrea/inducido químicamente , Diarrea/tratamiento farmacológico , Encefalina D-Penicilamina (2,5)/efectos adversos , Encefalina D-Penicilamina (2,5)/uso terapéutico , Fentanilo/efectos adversos , Fentanilo/uso terapéutico , Calor , Masculino , Morfina/efectos adversos , Morfina/uso terapéutico , Derivados de la Morfina/efectos adversos , Derivados de la Morfina/uso terapéutico , Oxicodona/efectos adversos , Oxicodona/uso terapéutico , Pirrolidinas/efectos adversos , Pirrolidinas/uso terapéutico , Ratas Sprague-Dawley , Insuficiencia Respiratoria/fisiopatologíaRESUMEN
Physical exercise stimulates the release of endogenous opioid peptides supposed to be responsible for changes in mood, anxiety, and performance. Exercise alters sensitivity to these effects that modify the efficacy at the opioid receptor. Although there is evidence that relates exercise to neuropeptide expression in the brain, the effects of exercise on opioid receptor binding and signal transduction mechanisms downstream of these receptors have not been explored. Here, we characterized the binding and G protein activation of mu opioid receptor, kappa opioid receptor or delta opioid receptor in several brain regions following acute (7 days) and chronic (30 days) exercise. As regards short- (acute) or long-term effects (chronic) of exercise, overall, higher opioid receptor binding was observed in acute-exercise animals and the opposite was found in the chronic-exercise animals. The binding of [(35) S]GTPγS under basal conditions (absence of agonists) was elevated in sensorimotor cortex and hippocampus, an effect more evident after chronic exercise. Divergence of findings was observed for mu opioid receptor, kappa opioid receptor, and delta opioid receptor receptor activation in our study. Our results support existing evidence of opioid receptor binding and G protein activation occurring differentially in brain regions in response to diverse exercise stimuli. We characterized the binding and G protein activation of mu, kappa, and delta opioid receptors in several brain regions following acute (7 days) and chronic (30 days) exercise. Higher opioid receptor binding was observed in the acute exercise animal group and opposite findings in the chronic exercise group. Higher G protein activation under basal conditions was noted in rats submitted to chronic exercise, as visible in the depicted pseudo-color autoradiograms.
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Química Encefálica , Condicionamiento Físico Animal , Receptores Opioides/metabolismo , Amígdala del Cerebelo/metabolismo , Animales , Bencenoacetamidas/metabolismo , Corteza Cerebral/metabolismo , Electrochoque , Encefalina D-Penicilamina (2,5)/metabolismo , Activación Enzimática , Proteínas de Unión al GTP/metabolismo , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Hipocampo/metabolismo , Hipotálamo/metabolismo , Naloxona/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Péptidos Opioides/metabolismo , Unión Proteica , Pirrolidinas/metabolismo , Ratas , Transducción de Señal , Factores de TiempoRESUMEN
Glucokinase activators (GKAs) are being developed for the treatment of type 2 diabetes. The toxicity of 4 GKAs (PF-04279405, PF-04651887, piragliatin, and PF-04937319) was assessed in mice, rats, dogs, and/or monkeys. GKAs were administered for 2 to 8 weeks. Standard endpoints, glucose, and insulin were assessed. All compounds produced varying degrees of hypoglycemia in all species. Brain neuronal necrosis and/or peripheral neuropathy were observed with most compounds. These findings are consistent with literature reports linking hypoglycemia with nervous system effects. Arteriopathy, mainly of cardiac vessels, was observed at a low frequency in monkey and/or dog. Arteriopathy occurred only at doses that produced severe and prolonged periods of repeated hypoglycemia. Since this lesion occurred in multiple studies with structurally distinct GKAs, these results suggested arteriopathy was related to GKA pharmacology. The morphological characteristics of the arteriopathy were consistent with that produced by experimental catecholamine administration. We hypothesize that the prolonged periods of hypoglycemia resulted in increased local and/or systemic concentrations of catecholamines via a counterregulatory and/or stress-related mechanism. Alternatively, prolonged hypoglycemia may have resulted in endothelial dysfunction leading to arteriopathy. This risk can be managed in human patients in clinical studies by careful glucose monitoring and intervention to avoid prolonged episodes of hypoglycemia.
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Azetidinas/efectos adversos , Bencenoacetamidas/efectos adversos , Benzofuranos/efectos adversos , Hipoglucemia/patología , Necrosis/patología , Enfermedades del Sistema Nervioso Periférico/patología , Pirimidinas/efectos adversos , Animales , Azetidinas/sangre , Bencenoacetamidas/sangre , Benzofuranos/sangre , Cromatografía Líquida de Alta Presión , Perros , Evaluación Preclínica de Medicamentos , Femenino , Hipoglucemia/inducido químicamente , Hipoglucemiantes/efectos adversos , Insulina/sangre , Macaca fascicularis , Masculino , Ratones , Ratones Endogámicos ICR , Necrosis/inducido químicamente , Neuronas/efectos de los fármacos , Neuronas/patología , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Pirimidinas/sangre , Ratas , Ratas Sprague-DawleyRESUMEN
Tuberculosis remains a global health emergency that calls for treatment regimens directed at new targets. Here we explored lipoamide dehydrogenase (Lpd), a metabolic and detoxifying enzyme in Mycobacterium tuberculosis (Mtb) whose deletion drastically impairs Mtb's ability to establish infection in the mouse. Upon screening more than 1.6 million compounds, we identified N-methylpyridine 3-sulfonamides as potent and species-selective inhibitors of Mtb Lpd affording >1000-fold selectivity versus the human homologue. The sulfonamides demonstrated low nanomolar affinity and bound at the lipoamide channel in an Lpd-inhibitor cocrystal. Their selectivity could be attributed, at least partially, to hydrogen bonding of the sulfonamide amide oxygen with the species variant Arg93 in the lipoamide channel. Although potent and selective, the sulfonamides did not enter mycobacteria, as determined by their inability to accumulate in Mtb to effective levels or to produce changes in intracellular metabolites. This work demonstrates that high potency and selectivity can be achieved at the lipoamide-binding site of Mtb Lpd, a site different from the NADâº/NADH pocket targeted by previously reported species-selective triazaspirodimethoxybenzoyl inhibitors.
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Antituberculosos/farmacología , Proteínas Bacterianas/antagonistas & inhibidores , Dihidrolipoamida Deshidrogenasa/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Mycobacterium tuberculosis/enzimología , Sulfonamidas/farmacología , Ácido Tióctico/análogos & derivados , Antituberculosos/efectos adversos , Antituberculosos/química , Arginina/química , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Bencenoacetamidas/efectos adversos , Bencenoacetamidas/química , Bencenoacetamidas/farmacología , Sitios de Unión , Transporte Biológico/efectos de los fármacos , Membrana Celular/metabolismo , Permeabilidad de la Membrana Celular , Dihidrolipoamida Deshidrogenasa/química , Dihidrolipoamida Deshidrogenasa/genética , Dihidrolipoamida Deshidrogenasa/metabolismo , Evaluación Preclínica de Medicamentos , Inhibidores Enzimáticos/efectos adversos , Inhibidores Enzimáticos/química , Ensayos Analíticos de Alto Rendimiento , Humanos , Moduladores del Transporte de Membrana/efectos adversos , Moduladores del Transporte de Membrana/química , Moduladores del Transporte de Membrana/farmacología , Pruebas de Sensibilidad Microbiana , Conformación Molecular , Proteínas Mutantes/antagonistas & inhibidores , Proteínas Mutantes/química , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Bibliotecas de Moléculas Pequeñas , Relación Estructura-Actividad , Sulfonamidas/efectos adversos , Sulfonamidas/química , Ácido Tióctico/metabolismoRESUMEN
Alzheimer's disease (AD) is the most common cause of dementia worldwide and mainly characterized by the aggregated ß-amyloid (Aß) and hyperphosphorylated tau. FLZ is a novel synthetic derivative of natural squamosamide and has been proved to improve memory deficits in dementia animal models. In this study, we aimed to investigate the mechanisms of FLZ's neuroprotective effect in APP/PS1 double transgenic mice and SH-SY5Y (APPwt/swe) cells. The results showed that treatment with FLZ significantly improved the memory deficits of APP/PS1 transgenic mice and decreased apoptosis of SH-SY5Y (APPwt/swe) cells. FLZ markedly attenuated Aß accumulation and tau phosphorylation both in vivo and in vitro. Mechanistic study showed that FLZ interfered APP processing, i.e., FLZ decreased ß-amyloid precursor protein (APP) phosphorylation, APP-carboxy-terminal fragment (APP-CTF) production and ß-amyloid precursor protein cleaving enzyme 1 (BACE1) expression. These results indicated that FLZ reduced Aß production through inhibiting amyloidogenic pathway. The mechanistic study about FLZ's inhibitory effect on tau phosphorylation revealed t the involvement of Akt/glycogen synthase kinase 3ß (GSK3ß) pathway. FLZ treatment increased Akt activity and inhibited GSK3ß activity both in vivo and in vitro. The inhibitory effect of FLZ on GSK3ß activity and tau phosphorylation was suppressed by inhibiting Akt activity, indicating that Akt/GSK3ß pathway might be the possible mechanism involved in the inhibitory effect of FLZ on tau hyperphosphorylation. These results suggested FLZ might be a potential anti-AD drug as it not only reduced Aß production via inhibition amyloidogenic APP processing pathway, but also attenuated tau hyperphosphoylation mediated by Akt/GSK3ß.
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Enfermedad de Alzheimer/tratamiento farmacológico , Péptidos beta-Amiloides/metabolismo , Bencenoacetamidas/farmacología , Fármacos Neuroprotectores/farmacología , Fenoles/farmacología , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Proteínas tau/metabolismo , Enfermedad de Alzheimer/metabolismo , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Animales , Apoptosis , Ácido Aspártico Endopeptidasas/metabolismo , Caspasa 3/metabolismo , Línea Celular Tumoral , Evaluación Preclínica de Medicamentos , Reacción de Fuga , Glucógeno Sintasa Quinasa 3/metabolismo , Glucógeno Sintasa Quinasa 3 beta , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Humanos , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Trastornos de la Memoria/tratamiento farmacológico , Trastornos de la Memoria/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de SeñalRESUMEN
The majority of anticancer therapeutics have failed to control the target cancers. Thus, new rational design concepts are critical. In most of the biological reactions, a cascade pathway is used to activate appropriate responses. In the cascade pathway, a small signal derived from neighboring environments can be amplified and it further triggers overwhelming and specialized responses. It can be applied to achieve powerful therapeutic effects for novel drug design strategies. Inspired by this concept, we design a preferential dual anti-cancer therapeutic cassette composed of (i) DNA/RNA nanostructures as both anticancer containers and target ligands and (ii) a gold nanocrystal as localized heat inducers. We demonstrate that this multi-modular platform is superior to conventional cancer medications in that it had higher drug loading efficiency, tunable drug release, and intrinsic serum stability characteristics. Both doxorubicin chemotherapy and thermal ablation exert a powerful synergistic killing effect that resulted in prostate cancer regression both in vitro and in vivo. We speculate that our novel anti-cancer drug system can be adapted to effectively destroy many different types of solid cancers.
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Luz , Nanopartículas/química , Animales , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/uso terapéutico , Antibióticos Antineoplásicos/toxicidad , Bencenoacetamidas/química , Línea Celular , Supervivencia Celular/efectos de los fármacos , ADN/química , Doxorrubicina/química , Doxorrubicina/uso terapéutico , Doxorrubicina/toxicidad , Diseño de Fármacos , Oro/química , Humanos , Hipertermia Inducida , Masculino , Nanopartículas del Metal/química , Nanopartículas del Metal/efectos de la radiación , Nanopartículas del Metal/uso terapéutico , Ratones , Ratones Desnudos , Piperidonas/química , Neoplasias de la Próstata/tratamiento farmacológico , ARN/química , Trasplante HeterólogoAsunto(s)
Antineoplásicos/uso terapéutico , Actitud del Personal de Salud , Bencenoacetamidas , Glutamina/análogos & derivados , Neoplasias/tratamiento farmacológico , Fenilacetatos , Piperidonas , Investigación Biomédica , Competencia Clínica , Ensayos Clínicos Fase III como Asunto , Combinación de Medicamentos , Humanos , Masculino , TexasRESUMEN
The incidence of fungal disease has increased dramatically over the past decades, mainly due to the emergence and transmission of antifungal resistance within the fungal pathogens. The present study investigates the use of novel antifungal compound 4-Phenyl-1-Napthyl Phenyl Acetamide (4P1NPA), isolated from marine Streptomyces sp. DPTB16 as a potent antifungal drug. The preclinical studies and molecular docking for 4P1NPA against Cytochrome P450 51 (CYP 51) were performed using in silico pharmacology and docking tools. The finding reveals the drug likeliness of 4P1NPA and satisfactory interaction of 4P1NPA with CYP 51. These results collectively evidence the use of 4P1NPA as a drug to treat fungal infections. On the whole, we highlight the findings of this research will be helpful to design 4P1NPA as novel antifungal drug to defend the emerging antifungal resistance.
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Antifúngicos/farmacología , Bencenoacetamidas/farmacología , Streptomyces/química , Animales , Antifúngicos/farmacocinética , Antifúngicos/toxicidad , Bencenoacetamidas/aislamiento & purificación , Bencenoacetamidas/farmacocinética , Inhibidores Enzimáticos del Citocromo P-450 , Evaluación Preclínica de Medicamentos , Ratones , Modelos Moleculares , Filogenia , RatasRESUMEN
It was reported previously that isolated human islets from individuals with type 2 diabetes mellitus (T2DM) show reduced glucose-stimulated insulin release. To assess the possibility that impaired bioenergetics may contribute to this defect, glucose-stimulated respiration (Vo(2)), glucose usage and oxidation, intracellular Ca(2+), and insulin secretion (IS) were measured in pancreatic islets isolated from three healthy and three type 2 diabetic organ donors. Isolated mouse and rat islets were studied for comparison. Islets were exposed to a "staircase" glucose stimulus, whereas IR and Vo(2) were measured. Vo(2) of human islets from normals and diabetics increased sigmoidally from equal baselines of 0.25 nmol/100 islets/min as a function of glucose concentration. Maximal Vo(2) of normal islets at 24 mM glucose was 0.40 ± 0.02 nmol·min(-1)·100 islets(-1), and the glucose S(0.5) was 4.39 ± 0.10 mM. The glucose stimulation of respiration of islets from diabetics was lower, V(max) of 0.32 ± 0.01 nmol·min(-1)·100 islets(-1), and the S(0.5) shifted to 5.43 ± 0.13 mM. Glucose-stimulated IS and the rise of intracellular Ca(2+) were also reduced in diabetic islets. A clinically effective glucokinase activator normalized the defective Vo(2), IR, and free calcium responses during glucose stimulation in islets from type 2 diabetics. The body of data shows that there is a clear relationship between the pancreatic islet energy (ATP) production rate and IS. This relationship was similar for normal human, mouse, and rat islets and the data for all species fitted a single sigmoidal curve. The shared threshold rate for IS was â¼13 pmol·min(-1)·islet(-1). Exendin-4, a GLP-1 analog, shifted the ATP production-IS curve to the left and greatly potentiated IS with an ATP production rate threshold of â¼10 pmol·min(-1)·islet(-1). Our data suggest that impaired ß-cell bioenergetics resulting in greatly reduced ATP production is critical in the molecular pathogenesis of type 2 diabetes mellitus.
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Bencenoacetamidas/farmacología , Diabetes Mellitus Tipo 2/metabolismo , Activadores de Enzimas/farmacología , Glucoquinasa/metabolismo , Glucosa/metabolismo , Insulina/metabolismo , Islotes Pancreáticos/efectos de los fármacos , Adulto , Animales , Señalización del Calcio/efectos de los fármacos , Respiración de la Célula/efectos de los fármacos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Exenatida , Femenino , Péptido 1 Similar al Glucagón/análogos & derivados , Glucoquinasa/química , Glucólisis/efectos de los fármacos , Humanos , Hipoglucemiantes/farmacología , Secreción de Insulina , Islotes Pancreáticos/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Fosforilación Oxidativa/efectos de los fármacos , Péptidos/farmacología , Ratas , Especificidad de la Especie , Técnicas de Cultivo de Tejidos , Ponzoñas/farmacologíaRESUMEN
PURPOSE: Most of postoperative patients experience pain, state anxiety and sleep disturbance. These problems negatively influence the recovery of postoperative patients. So alleviating these problems has been one of the nurses' central roles. The purpose of this study was to examine the effects of back massage on pain, state anxiety and quality of sleep of postoperative gastrectomy patients. METHODS: A non-synchronized non-equivalent control group pre and post-test design was used. The research instruments used in this study were the Numerical Rating Scale (NRS) for pain, the State-Anxiety Inventory (STAI) for anxiety and the Verran and Synder-Halpern scale for quality of sleep. The subjects were patients admitted to a university hospital located in D city. Twenty-nine patients in the experimental group had a 10 min manual back massage stimulation for 5 days from the 1st day to the 5th day after their operation, and 25 patients in the control group did not. RESULTS: The degree of pain was significantly reduced according to post operation day and quality of sleep was significantly increased. However state anxiety was not significantly reduced. CONCLUSION: Back massage is a partially effective nursing intervention for postoperative patients with gastrectomy who experience pain and sleep disturbance.
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Humanos , Ansiedad , Bencenoacetamidas , Gastrectomía , Masaje , PiperidonasRESUMEN
Two-pore domain potassium (K(2P)) channels play a key role in setting the membrane potential of excitable cells. Despite their role as putative targets for drugs and general anesthetics, little is known about the structure and the drug binding site of K(2P) channels. We describe A1899 as a potent and highly selective blocker of the K(2P) channel TASK-1. As A1899 acts as an open-channel blocker and binds to residues forming the wall of the central cavity, the drug was used to further our understanding of the channel pore. Using alanine mutagenesis screens, we have identified residues in both pore loops, the M2 and M4 segments, and the halothane response element to form the drug binding site of TASK-1. Our experimental data were used to validate a K(2P) open-pore homology model of TASK-1, providing structural insights for future rational design of drugs targeting K(2P) channels.