RESUMEN

Oxidative stress plays a central role in cataract formation suggesting that antioxidants might slow cataract progression. The anticataract activity of N-acetylcysteine amide (NACA) and (2 R, 2 R')-3,3'-disulfanediyl bis(2-acetamidopropanamide) (diNACA) and/or N-acetylcysteine (NAC), were evaluated in porcine and rat lens models. Cataractogenesis via oxidation was induced with H2O2 and/or glucose oxidase (GO). Porcine lenses were incubated in 0.1 mM, 1 mM, or 10 mM NAC, NACA or diNACA for 24 h. Lenses were then transferred to media containing 0.75 mM H2O2 and 4.63U of GO in order to maintain a constant H2O2 level for an additional 8 h. At the end of incubation, lenses were imaged under darkfield microscopy. Separately, rat lenses were extracted from 3-week-old Wistar rats and incubated with either 10 mM NACA or 10 mM diNACA for 24 h prior to treatment with 0.2U GO to generate a steady source of ∼0.6 mM H2O2. Rat lenses were analyzed by LC-MS/MS to quantify changes in cysteine, cystine, glutathione (GSH) or oxidised glutathione (GSSG) levels in the lens epithelium, cortex or core. Pre-treatment with NACA or diNACA followed by oxidation with H2O2 and/or GO to stimulate cataract formation afforded rapid assessment in ex vivo porcine (32 h) and rat (48 h) lens models. Pre-treatment of isolated porcine lenses with 0.1 mM, 1 mM or 10 mM of either NAC, NACA or diNACA followed by H2O2/GO treatment resulted in reduced lens opacity relative to the lenses exposed to H2O2/GO, with NACA and diNACA reducing opacities to a greater extent than NAC. Rat lenses incubated with 10 mM NACA or 10 mM diNACA without exposure to H2O2 showed no signs of opacities. Pre-treatment of rat lenses with 10 mM NACA or 10 mM diNACA, followed by GO cataract induction resulted in reduced opacities compared to control (GO alone). LC-MS/MS analyses revealed that NACA, but not diNACA, increased cysteine, cystine and GSH levels in rat lens epithelium and cortex regions. Taken together, both NACA and diNACA inhibited cataract formation to a greater extent than NAC (all at 1-10 mM) in an ex vivo porcine lens model. Both NACA and diNACA (both at 10 mM) reduced cataract formation in rat lenses. Based on LC-MS/MS analyses, NACA-induced reduction in opacity observed in rat lenses was attributed to enhanced cysteine and GSH levels while the diNACA-induced reduction in opacity induced did not consistently increase cysteine, cystine and GSH levels and, therefore, appears to involve a different antioxidant mechanism. These screening studies warrant further testing of NACA and diNACA as anticataract agents.

Asunto(s)

Catarata , Cristalino , Ratas , Animales , Porcinos , Acetilcisteína/efectos adversos , Peróxido de Hidrógeno/farmacología , Cistina/efectos adversos , Cromatografía Liquida , Ratas Wistar , Espectrometría de Masas en Tándem , Cristalino/metabolismo , Catarata/inducido químicamente , Antioxidantes , Estrés Oxidativo , Glutatión/metabolismo , Proteínas , Disulfuro de Glutatión

RESUMEN

SRS27, an andrographolide analogue, had been proven to have therapeutic properties at a dose of 3 mg/kg in both in vitro and in vivo asthma models of our previous study. The present study focuses on the pharmacokinetic and toxicity profile of this compound to provide further evidence for the development of this compound as an anti-asthma agent. A simple pharmacokinetic study was performed in female BALB/c mice to measure blood plasma concentration of the compound at therapeutic dose. At a single dose of 3 mg/kg, SRS27 had a relatively short half-life but was able to achieve a concentration range of 13-19 µM that is related to its in vitro bioactivities. With regard to toxicity profile, SRS27 appears to be safe, as no histopathological changes were observed in the liver, kidneys and ovaries of SRS27-treated female BALB/c mice. In addition, there was no significant change in the mean body weight and organ weight of the animals in the SRS27-treated groups compared with the vehicle-treated control group at the end of the treatment. This fully supports the absence of any significant changes in peripheral blood leukocyte counts of SRS27-treated mice. Rewardingly, this acute toxicity study also revealed that SRS27 has a wide therapeutic window as no toxicity symptoms were detected with a dose up to 60 mg/kg daily when tested for 14 days. These results provide strong justification for further investigation of SRS27 as a potential new anti-asthma agent.

Asunto(s)

Antiasmáticos/farmacocinética , Antiasmáticos/toxicidad , Diterpenos/farmacocinética , Diterpenos/toxicidad , Lactonas/farmacocinética , Lactonas/toxicidad , Animales , Antiasmáticos/sangre , Disponibilidad Biológica , Diterpenos/sangre , Femenino , Riñón/anatomía & histología , Riñón/efectos de los fármacos , Lactonas/sangre , Hígado/anatomía & histología , Hígado/efectos de los fármacos , Pulmón/anatomía & histología , Pulmón/efectos de los fármacos , Ratones Endogámicos BALB C , Ovario/anatomía & histología , Ovario/efectos de los fármacos

RESUMEN

Andrographolide (AGP) and 14-deoxy-11,12-didehydroandrographolide (DDAG), two main diterpenoid constituents of Andrographis paniculata were previously shown to ameliorate asthmatic symptoms in a mouse model. However, due to inadequacies of both compounds in terms of drug-likeness, DDAG analogues were semisynthesised for assessment of their anti-asthma activity. A selected analogue, 3,19-diacetyl-14-deoxy-11,12-didehydroandrographolide (SRS27), was tested for inhibitory activity of NF-κB activation in TNF-α-induced A549 cells and was subsequently evaluated in a mouse model of ovalbumin (OVA)-induced asthma. Female BALB/c mice, 6-8weeks old were sensitized on days 0 and 14, and challenged on days 22, 23 and 24 with OVA. Compound or vehicle (3% dimethyl sulfoxide) was administered intraperitoneally 1h before and 11h after each OVA aerosol challenge. On day 25, pulmonary eosinophilia, airway hyperresponsiveness, mucus hypersecretion, inflammatory cytokines such as IL-4, -5 and -13 in BAL fluid, gene expression of inflammatory mediators such as 5-LOX, E-selectin, VCAM-1, CCL5, TNF-α, AMCase, Ym2, YKL-40, Muc5ac, CCL2 and iNOS in animal lung tissues, and serum IgE were determined. SRS27 at 30µM was found to suppress NF-κB nuclear translocation in A549 cells. In the ovalbumin-induced mouse asthma model, SRS27 at 3mg/kg displayed a substantial decrease in pulmonary eosinophilia, BAL fluid inflammatory cytokines level, serum IgE production, mucus hypersecretion and gene expression of inflammatory mediators in lung tissues. SRS27 is the first known DDAG analogue effective in ameliorating inflammation and airway hyperresponsiveness in the ovalbumin-induced mouse asthma model.

Asunto(s)

Antiasmáticos/uso terapéutico , Antiinflamatorios/uso terapéutico , Asma/tratamiento farmacológico , Diterpenos/uso terapéutico , Lactonas/uso terapéutico , FN-kappa B/antagonistas & inhibidores , Células A549 , Animales , Antiasmáticos/farmacología , Antiinflamatorios/farmacología , Asma/sangre , Asma/inmunología , Asma/fisiopatología , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Recuento de Células , Citocinas/inmunología , Modelos Animales de Enfermedad , Diterpenos/farmacología , Femenino , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Lactonas/farmacología , Ratones Endogámicos BALB C , FN-kappa B/genética , FN-kappa B/inmunología , Ovalbúmina , Transducción de Señal

RESUMEN

INTRODUCTION: The traditional Kocher approach for lateral radial head exposure may be complicated by injury to the deep branch of the radial nerve (DBRN) and the radial collateral ligament. Kaplan approach is less commonly used, due to its known proximity to the DBRN. Extensor Digitorum Communis (EDC) splitting approach allows possible wide surgical exposure and low risk of radial collateral ligament injury. The comparison of the proximity of the DBRN to the surgical dissection at the level of radial head among approaches to the radial head has not previously been evaluated. We aimed to determine the anatomical proximity of the DBRN in these 3 common radial head approaches and to define a safe zone of dissection for the surgical exposure. METHODS: Cadaveric dissections of 9 pairs of fresh frozen upper extremities were performed using EDC splitting, Kaplan and Kocher approach to the radial head sequentially in a randomized order. A mark was made on the radial head upon initial exposure during dissection. Measurements from the marked point of the radial head to the DBRN were made at the level of radial head. RESULTS: The distance of DBRN to the radial head was 20 (17-22) mm in EDC splitting approach, 7 (3-11) mm in Kaplan approach and 29 (25-33) mm in Kocher approach. The EDC splitting approach was associated with a significantly lower chance of encountering the DBRN at the level of radial head as compared to the Kaplan approach (P<0.001). In all cases, lateral ligamentous complex was not exposed in Kaplan and EDC approaches, but were encountered in Kocher approach, risking injury to the radial collateral ligament. CONCLUSIONS: The EDC splitting approach provides adequate exposure without the need to elevate or retract the EDC and ECU muscle mass that could risk injuring the DBRN. The Kaplan approach should be done by experienced surgeons who are familiar with the anatomy in this region, with extreme caution due to proximity of the DBRN to the surgical dissection at the level of the radial head. Caution of the DBRN should be taken during anterior elevation and retraction of the muscle mass in Kocher approach. LEVEL OF EVIDENCE: IV.

Asunto(s)

Disección/métodos , Complicaciones Intraoperatorias/prevención & control , Procedimientos Ortopédicos/métodos , Traumatismos de los Nervios Periféricos/prevención & control , Nervio Radial/anatomía & histología , Radio (Anatomía)/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Traumatismos de los Nervios Periféricos/etiología , Nervio Radial/lesiones , Nervio Radial/cirugía , Radio (Anatomía)/inervación

RESUMEN

There is broad agreement among health-care stakeholders that more must be done to ensure that patients have timely access to new and innovative medicines. Assuming that industry will continue to develop such medicines at a sustainable rate, regulators and payers become the gatekeepers. Regulators, starting in the late 1980s/early 1990s, and, more recently, payers have implemented a variety of early-access pathways or initiatives, and this practice is continuing even today. This article describes the specific approaches that have been taken in four economically developed regions, reviews their success rates, and suggests possible new directions.

Asunto(s)

Accesibilidad a los Servicios de Salud , Necesidades y Demandas de Servicios de Salud , Preparaciones Farmacéuticas/provisión & distribución , Tecnología Biomédica , Canadá , Humanos , Mecanismo de Reembolso , Singapur , Estados Unidos , United States Food and Drug Administration

RESUMEN

OBJECTIVES: Micro-CT provides three-dimensional details and has been widely used for biomedical assessments. This study aimed to determine the most appropriate threshold method for quantitatively assessing the dynamics of periodontal destruction. METHODS: Inflammation was induced by submerging a silk ligature in the sulcus of the maxillary second molars of rats, and the animals were killed prior to ligature placement and after 7 and 21 days. The maxillae were examined for the bone resorptive activities by micro-CT, histology and tartrate-resistant acid phosphatase staining. The imaging threshold was determined by CT phantom, global and local algorithms. A bone fraction measurement from each threshold-determining technique was compared with histomorphometry. The reliability and reproducibility were examined by the intraclass correlation coefficient (ICC) and the coefficient of variation. RESULTS: Significant reduction of inflammatory infiltration (p < 0.01) and active osteoclastic resorption (p < 0.05) from Day 7 to Day 21 were noted. High inter- and intraexaminer agreement were demonstrated in both histomorphometric and micro-CT assessments (ICC > 0.98). The algorithm-based technique demonstrated stronger correlation to histomorphometry than phantom-based thresholds, and the highest agreement was presented by the local algorithm (ICC > 0.96). This, however, was considerably computationally expensive. CONCLUSIONS: The local threshold-determining algorithm is suggested for examining inflammation-induced bone loss. Further investigation will be aimed at enhancing computational efficiency.

Asunto(s)

Pérdida de Hueso Alveolar/diagnóstico por imagen , Procesamiento de Imagen Asistido por Computador/métodos , Microtomografía por Rayos X/métodos , Fosfatasa Ácida/análisis , Algoritmos , Pérdida de Hueso Alveolar/patología , Animales , Biomarcadores/análisis , Colágeno , Colorantes , Tejido Conectivo/patología , Modelos Animales de Enfermedad , Encía/patología , Imagenología Tridimensional/métodos , Isoenzimas/análisis , Masculino , Enfermedades Maxilares/diagnóstico por imagen , Enfermedades Maxilares/patología , Diente Molar/diagnóstico por imagen , Osteoclastos/patología , Periodontitis/diagnóstico por imagen , Periodontitis/patología , Fantasmas de Imagen , Proyectos Piloto , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Fosfatasa Ácida Tartratorresistente , Factores de Tiempo , Cuello del Diente/patología

RESUMEN

Genetic changes in HER2, PTEN, PIK3CA and AKT1 are all common in breast cancer and lead to the elevated phosphorylation of downstream targets of the PI3K/AKT signalling pathway. Changes in HER2, PTEN, PIK3CA and AKT have all been reported to lead to both enhanced proliferation and failures in hollow lumen formation in three dimensional epithelial culture models, but it is not clear whether these failures in lumen formation are caused by any failure in the spatial coordination of lumen formation (hollowing) or purely a failure in the apoptosis and clearance of luminal cells (cavitation). Here, we use normal murine mammary gland (NMuMG) epithelial cells, which form a hollow lumen without significant apoptosis, to compare the transformation by these four genetic changes. We find that either mutant PIK3CA expression or PTEN loss, but not mutant AKT1 E17K, cause disrupted epithelial architecture, whereas HER2 overexpression drives strong proliferation without affecting lumen formation in these cells. We also show that PTEN requires both lipid and protein phosphatase activity, its extreme C-terminal PDZ binding sequence and probably Myosin 5A to control lumen formation through a mechanism that does not correlate with its ability to control AKT, but which is selectively lost through mutation in some tumours. These findings correlate AKT-independent signalling activated by mutant PIK3CA or PTEN loss, but not strongly by HER2, with disrupted epithelial architecture and tumour formation.

Asunto(s)

Fosfatidilinositol 3-Quinasa Clase Ia/genética , Células Epiteliales/metabolismo , Células Epiteliales/patología , Eliminación de Gen , Mutación , Fosfohidrolasa PTEN/genética , Proteínas Proto-Oncogénicas c-akt/genética , Receptor ErbB-2/genética , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/metabolismo , Cromonas/farmacología , Fosfatidilinositol 3-Quinasa Clase Ia/metabolismo , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Glándulas Mamarias Humanas/metabolismo , Glándulas Mamarias Humanas/patología , Morfolinas/farmacología , Fosfohidrolasa PTEN/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3 , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptor ErbB-2/metabolismo

RESUMEN

Signaling by extracellular purines such as ATP and adenosine has implications for dental research on multiple levels, with the association of purinergic signaling with inflammation, mechanical strain, and pain making the system particularly relevant for the specific challenges in the oral cavity. Oral tissues express a variety of G-protein-coupled P2Y receptors for ATP and P1 receptors for adenosine in addition to ionotropic P2X receptors for ATP. When these receptors are combined with the plethora of extracellular enzymes capable of manipulating extracellular agonist levels, a complex system for regulating oral health emerges, and recent findings have begun to identify a key role for purinergic signaling in oral pathophysiology. For example, the manipulation of extracellular ATP levels by P. gingivalis reduces inflammasome activation and apoptosis linked to P2X(7) receptor activation. Release of ATP by periodontal ligaments may link mechanical strain to bone remodeling. Activation of P2X receptors is implicated in dental pain, and receptor antagonists represent important targets for new analgesics. Altered levels of adenosine receptors in periodontal disease also suggest a role for nucleosides in dental signaling. The intricacies of the purinergic signaling system make it well-suited for the unique concerns of dental research, and future findings will doubtless confirm this importance.

Asunto(s)

Enfermedades de la Boca/fisiopatología , Dolor/fisiopatología , Receptores Purinérgicos P2X/fisiología , Receptores Purinérgicos P2Y/fisiología , Transducción de Señal/fisiología , Adenosina Trifosfato/metabolismo , Animales , Humanos , Inflamasomas/fisiología , Inflamación/metabolismo , Boca/fisiología

RESUMEN

Traditional drug licensing approaches are based on binary decisions. At the moment of licensing, an experimental therapy is presumptively transformed into a fully vetted, safe, efficacious therapy. By contrast, adaptive licensing (AL) approaches are based on stepwise learning under conditions of acknowledged uncertainty, with iterative phases of data gathering and regulatory evaluation. This approach allows approval to align more closely with patient needs for timely access to new technologies and for data to inform medical decisions. The concept of AL embraces a range of perspectives. Some see AL as an evolutionary step, extending elements that are now in place. Others envision a transformative framework that may require legislative action before implementation. This article summarizes recent AL proposals; discusses how proposals might be translated into practice, with illustrations in different therapeutic areas; and identifies unresolved issues to inform decisions on the design and implementation of AL.

Asunto(s)

Aprobación de Drogas/legislación & jurisprudencia , Aprobación de Drogas/métodos , Necesidades y Demandas de Servicios de Salud/legislación & jurisprudencia , Necesidades y Demandas de Servicios de Salud/organización & administración , Concesión de Licencias/legislación & jurisprudencia , Animales , Toma de Decisiones , Unión Europea , Humanos , Estados Unidos

RESUMEN

The endocannabinoid system (ECS) is activated at the onset of obesity and diverse metabolic diseases. Endocannabinoids mediate their physiological and behavioral effects by activating specific cannabinoid receptors, mainly cannabinoid receptor 1 (CB(1)R). Diabetic nephropathy (DN) is induced by hyperlipidemia, and renal proximal tubule cells are an important site for the onset of DN. However, the pathophysiology of CB(1)R, especially in the hyperlipidemia of DN, has not been elucidated. Therefore, we examined the effect of palmitic acid (PA) on CB(1)R expression and its related signal pathways in human renal proximal tubular cells (HK-2 cells). PA significantly increased CB(1)R mRNA and protein levels and induced CB(1)R internalization. PA-induced activation of CB(1)R is prevented by the treatment of AACOCF(3) (a cPLA(2) inhibitor), indomethacin and NS398 (a COX 2 inhibitors). Indeed, PA increased cPLA(2), and COX-2 but not COX-1. We also investigated whether the PA-induced activation of CB(1)R is linked to apoptosis. As a result, AM251 (a CB(1)R antagonist) attenuated PA-mediated apoptosis in a concentration-dependent manner. Furthermore, PA decreased GRP78 expression and induced increases in the endoplasmic reticulum (ER) stress signaling pathways p-PERK, p-eIF2α, p-ATF4, and CHOP, which were blocked by AM251 treatment. Moreover, PA increased the Bax/Bcl-2 ratio, cleaved PARP, and caspase-3 levels. The PA-induced apoptotic effects were decreased with CB(1)R-specific antagonist (AM251) treatment and CB1 si-RNA transfection. In conclusion, PA induced apoptosis through ER stress via CB(1)R expression in human proximal tubule cells. Our results provide evidence that CB(1)R blockade may be a potential anti-diabetic therapy for the treatment of DN.

Asunto(s)

Apoptosis , Nefropatías Diabéticas/metabolismo , Retículo Endoplásmico/metabolismo , Túbulos Renales Proximales/metabolismo , Ácido Palmítico/metabolismo , Receptor Cannabinoide CB1/metabolismo , Transducción de Señal , Estrés Fisiológico , Factor de Transcripción Activador 4/metabolismo , Apoptosis/efectos de los fármacos , Caspasa 3/metabolismo , Proliferación Celular , Inhibidores de la Ciclooxigenasa 2/farmacología , Nefropatías Diabéticas/etiología , Nefropatías Diabéticas/patología , Relación Dosis-Respuesta a Droga , Endocitosis , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/patología , Chaperón BiP del Retículo Endoplásmico , Inhibidores Enzimáticos/farmacología , Factor 2 Eucariótico de Iniciación/metabolismo , Proteínas de Choque Térmico/metabolismo , Humanos , Hiperlipidemias/complicaciones , Hiperlipidemias/metabolismo , Túbulos Renales Proximales/efectos de los fármacos , Túbulos Renales Proximales/patología , Fosfolipasas A2 Citosólicas/antagonistas & inhibidores , Fosfolipasas A2 Citosólicas/metabolismo , Fosforilación , Poli(ADP-Ribosa) Polimerasas/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Interferencia de ARN , ARN Mensajero/metabolismo , Receptor Cannabinoide CB1/efectos de los fármacos , Receptor Cannabinoide CB1/genética , Transducción de Señal/efectos de los fármacos , Estrés Fisiológico/efectos de los fármacos , Factores de Tiempo , Factor de Transcripción CHOP/metabolismo , Proteína X Asociada a bcl-2/metabolismo , eIF-2 Quinasa/metabolismo

RESUMEN

PURPOSE: To characterize the expression patterns of the Na+-K+-Cl(-) cotransporter (NKCC) 1 and NKCC2, and the Na+-Cl(-) cotransporter (NCC) in the rat lens and to determine if they play a role in regulating lens volume and transparency. METHODS: RT-PCR was performed on RNA extracted from fiber cells to identify sodium dependent cotransporters expressed in the rat lens. Western blotting and immunohistochemistry, using NKCC1, NKCC2, and NCC antibodies, were used to verify expression at the protein level and to localize transporter expression. Organ cultured rat lenses were incubated in Artificial Aqueous Humor (AAH) of varying osmolarities or isotonic AAH that contained either the NKCC specific inhibitor bumetanide, or the NCC specific inhibitor thiazide for up to 18 h. Lens transparency was monitored with dark field microscopy, while tissue morphology and antibody labeling patterns were recorded using a confocal microscope. RESULTS: Molecular experiments showed that NKCC1 and NCC were expressed in the lens at both the transcript and protein levels, but NKCC2 was not. Immunohistochemistry showed that both NKCC1 and NCC were expressed in the lens cortex, but NCC expression was also found in the lens core. In the lens cortex the majority of labeling for both transporters was cytoplasmic in nature, while in the lens core, NCC labeling was associated with the membrane. Exposure of lenses to either hypotonic or hypertonic AAH had no noticeable effects on the predominantly cytoplasmic location of either transporter in the lens cortex. Incubation of lenses in isotonic AAH plus the NKCC inhibitor bumetanide for 18 h induced a cortical opacity that was initiated by a shrinkage of peripheral fiber cells and the dilation of the extracellular space between fiber cells in a deeper zone located some approximately 150 microm in from the capsule. In contrast, lenses incubated in isotonic AAH and the NCC inhibitor thiazide maintained both their transparency and their regular fiber cell morphology. CONCLUSIONS: We have confirmed the expression of NKCC1 in the rat lens and report for the first time the expression of NCC in lens fiber cells. The expression patterns of the two transporters and the differential effects of their specific inhibitors on fiber cell morphology indicate that these transporters play distinct roles in the lens. NKCC1 appears to mediate ion influx in the lens cortex while NCC may play a role in the lens nucleus.

Asunto(s)

Cristalino/fisiología , Simportadores del Cloruro de Sodio/metabolismo , Simportadores de Cloruro de Sodio-Potasio/metabolismo , Animales , Western Blotting , Bumetanida/farmacología , Homeostasis/fisiología , Inmunohistoquímica , Corteza del Cristalino/metabolismo , Cristalino/citología , Técnicas de Cultivo de Órganos , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Simportadores del Cloruro de Sodio/efectos de los fármacos , Inhibidores del Simportador de Cloruro Sódico y Cloruro Potásico/farmacología , Simportadores de Cloruro de Sodio-Potasio/efectos de los fármacos , Miembro 1 de la Familia de Transportadores de Soluto 12 , Miembro 2 de la Familia de Transportadores de Soluto 12 , Tiazidas/farmacología , Distribución Tisular

RESUMEN

Solanum nigrum L. (SNL) has been traditionally used as a herbal plant, whose fruit is believed to have anti-tumor properties, although the mechanism for the activity remains to be elucidated. In this study, we prepared an ethanol extract from ripe fruits of SNL and investigated the mechanism involved in its growth-inhibitory effect on MCF-7 human breast cancer cells. Results from proliferation assay using tritium uptake showed that the proliferative capacity of MCF-7 cells was strongly suppressed in the presence of SNL ethanol extract. This was further confirmed through MTT assay and trypan blue exclusion experiments, which showed a very close correlation between the SNL extract concentration and the surviving cell numbers. The SNL extract-mediated suppression of cell growth was verified to be apoptotic, based on the appearance of DNA laddering, increase in DNA fragmentation, and low fluorescence intensity in nuclei after propidium iodide staining of the cells. Furthermore, the SNL extract was revealed to be a potential scavenger of hydroxyl radicals and DPPH radicals rather than superoxide anions. Collectively, our findings suggest that SNL fruit extract could be used as an anti-oxidant and cancer chemo-preventive material.

Asunto(s)

Apoptosis/efectos de los fármacos , Solanum nigrum/química , Antioxidantes/análisis , Antioxidantes/química , Compuestos de Bifenilo , Neoplasias de la Mama/patología , Supervivencia Celular/efectos de los fármacos , Fragmentación del ADN/efectos de los fármacos , ADN de Neoplasias/biosíntesis , ADN de Neoplasias/genética , Femenino , Citometría de Flujo , Radicales Libres/metabolismo , Humanos , Radical Hidroxilo/análisis , Fenoles/química , Picratos/química , Superóxidos/química , Células Tumorales Cultivadas

Asunto(s)

Anemia/etiología , Médula Ósea/patología , Núcleo Celular/patología , Mieloma Múltiple/patología , Anciano , Anciano de 80 o más Años , Anemia/patología , Biopsia , Núcleo Celular/ultraestructura , Diagnóstico Diferencial , Epoetina alfa , Eritropoyetina/uso terapéutico , Femenino , Filgrastim , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Humanos , Leucopenia , Mieloma Múltiple/ultraestructura , Proteínas Recombinantes

RESUMEN

Nucleotide sequence extending 2,3-dihydroxybiphenyl 1,2-dioxygenase gene (pcbC) and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase gene (pcbD) of Pseudomonas sp. DJ-12 was previously analyzed and the two genes were present in the order of pcbD-pcbC preceded by a promoter from Pseudomonas sp. DJ-12. In this study, a 3.8-kb nucleotide sequence located downstream of the pcbC gene was analyzed to have three open reading frames (ORFs) that are designated as orf1, pcbE and orf2 genes. All of the ORFs were preceded by each ribosome-binding sequence of 5-GGAXA-3 (X=G or A). However, no promoter-like sequence and transcription terminator sequence were found in the analyzed region, downstream of pcbC gene. Therefore, the gene cluster appeared to be present in the order of pcbD-pcbC-orf1-pcbE-orf2 as an operon, which is unique organization characterized so far in biphenyl- and PCB-degrading bacteria. The orf1 gene was composed of 1,224 base pairs which can encode a polypeptide of molecular weight 44,950 containing 405 amino acid residues. A deduced amino acid sequence of the orf1 gene product exhibited 21-33% identity with those of indole dioxygenase and phenol hydroxylase components. The pcbE gene was composed of 783 base pairs encoding 2-hydroxypenta-2,4-dienoate hydratase involved in the 4-chlorobiphenyl catabolism. The orf2 gene was composed of 1,017 base pairs encoding a polypeptide of molecular weight 37,378 containing 338 amino acid residues. A deduced amino acid sequence of the orf2 gene product exhibited 31% identity with that of a nitrilotriacetate monooxygenase component.

Asunto(s)

Genes Bacterianos/genética , Hidroliasas/genética , Pseudomonas/enzimología , Pseudomonas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Fenómenos Químicos , Química Física , Clonación Molecular , ADN Bacteriano/genética , Electroforesis en Gel de Poliacrilamida , Datos de Secuencia Molecular , Peso Molecular , Familia de Multigenes , Plásmidos , Espectrofotometría Ultravioleta

RESUMEN

BACKGROUND: Ebastine and loratadine are 2 nonsedating second-generation H(1) antihistamines with once-daily dosing. OBJECTIVE: We compared the efficacy and safety of ebastine 20 mg and 10 mg, loratadine 10 mg, and placebo administered once daily for 4 weeks in controlling the symptoms of seasonal allergic rhinitis (SAR). METHODS: In a double-blind, placebo-controlled, randomized, parallel-group study, 565 patients with ragweed SAR, ages 12 to 70 years, received either ebastine 20 mg, ebastine 10 mg, loratadine 10 mg, or placebo once daily for 4 weeks. Patients recorded morning and evening reflective scores (past 12 hours) as well as snapshot scores (at time of recording) for nasal discharge, congestion, sneezing, itching, and total eye symptoms. Total symptom score (TSS) is the sum of these 5 scores. RESULTS: Ebastine 20 mg produced significantly greater (P <.05) reductions from baseline compared with loratadine 10 mg over the entire treatment period in the mean daily reflective (42.5% vs 36.3%) and mean morning snapshot (40.3% vs 31.3%) TSS. The overall improvement in daily reflective and morning snapshot TSS was comparable between ebastine 10 mg and loratadine 10 mg and significantly better than placebo (P <.05). The total percent of patients with adverse events was similar among all 4 treatment groups (P =.78). CONCLUSION: Ebastine 20 mg given once daily was significantly superior to loratadine 10 mg given once daily at improving the rhinitis total symptom score throughout the day and at awakening over a 4-week period. Ebastine 20 mg and 10 mg doses were both efficacious and well tolerated in the treatment of SAR.

Asunto(s)

Butirofenonas/administración & dosificación , Butirofenonas/uso terapéutico , Antagonistas de los Receptores Histamínicos H1/administración & dosificación , Antagonistas de los Receptores Histamínicos H1/uso terapéutico , Loratadina/administración & dosificación , Loratadina/uso terapéutico , Piperidinas/administración & dosificación , Piperidinas/uso terapéutico , Rinitis Alérgica Estacional/tratamiento farmacológico , Adolescente , Adulto , Anciano , Niño , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad

Asunto(s)

Albuterol/análogos & derivados , Antiinflamatorios/administración & dosificación , Asma/tratamiento farmacológico , Broncodilatadores/administración & dosificación , Triamcinolona Acetonida/administración & dosificación , Adulto , Albuterol/administración & dosificación , Asma/fisiopatología , Quimioterapia Combinada , Femenino , Humanos , Masculino , Xinafoato de Salmeterol , Resultado del Tratamiento

RESUMEN

We report the case of a 57-year-old patient with a presumed developmental anomaly of the medial orbital wall. The resultant protrusion of orbital contents into the ethmoidal complex was clearly demonstrated on coronal computed tomography (CT) scans of the paranasal sinuses. This anomaly presents a high risk of iatrogenic injury to the medial rectus and orbit during functional endoscopic sinus surgery and has not previously been described.

Asunto(s)

Senos Etmoidales/diagnóstico por imagen , Obstrucción Nasal/diagnóstico por imagen , Órbita/anomalías , Endoscopía/efectos adversos , Femenino , Humanos , Persona de Mediana Edad , Obstrucción Nasal/etiología , Órbita/diagnóstico por imagen , Órbita/lesiones , Tomografía Computarizada por Rayos X

RESUMEN

Catechol 2,3-dioxygenase (C23O), one of extradiol-type dioxygenases cleaving aromatic C-C bond at meta position of dihydroxylated aromatic substrates, catalyzes the conversion of catechol to 2-hydroxymuconic semialdehyde. As our ongoing study to characterize biochemical and genetic properties of the extradiol-type dioxygenases at molecular level, a C23O gene encoded in chromosomal DNA of Alcaligenes eutrophus 335, a strain degrading phenol and p-cresol, was cloned. The C23O gene was localized in an 1.4-kb PstI fragment from A. eutrophus 335, and was expressed in E. coli HB101. The C23O exhibited the highest aromatic ring-fission activity to catechol as a substrate, and its relative activity to other dihydroxylated aromatic substrates was in order of catechol >> 4-methylcatechol > 3-methylcatechol, protocatechuate, 4-chlorocatechol > 3,4-dihydroxy-phenylacetate > 2,3-dihydroxybiphenyl. Nucleotide sequence of the 1.4-kb fragment has revealed that an open reading frame (ORF) corresponding to the C23O gene was composed of 930 base pairs. A putative ribosome-binding sequence of AGGAG was found at about 10 nucleotides upstream the ORF which can encode a polypeptide of molecular weight 34 kDa consisting of 309 amino acid residues. The deduced amino acid sequence of C23O from A. eutrophus 335 exhibited the highest 59% identity with those of corresponding enzymes from Pseudomonas sp. CF600 (p VI150), P. putida HS1 (pDK1), and P. putida PpG7 (NAH7). An alignment of amino acid sequences of extradiol-type dioxygenases including C23O from A. eutrophus 335 has revealed that catalytically and structurally important amino acid residues of the enzymes were conserved during evolution.

Asunto(s)

Alcaligenes/enzimología , Dioxigenasas , Oxigenasas/genética , Alcaligenes/genética , Secuencia de Aminoácidos , Secuencia de Bases , Catecol 2,3-Dioxigenasa , Clonación Molecular , ADN Bacteriano/química , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Alineación de Secuencia

Asunto(s)

Cefalosporinas/síntesis química , Cefalosporinas/farmacología , Animales , Cefalosporinas/farmacocinética , Pruebas de Sensibilidad Microbiana , Ratas , Relación Estructura-Actividad

Asunto(s)

Cefalosporinas/síntesis química , Cefalosporinas/farmacología , Animales , Cefalosporinas/farmacocinética , Pruebas de Sensibilidad Microbiana , Ratas , Relación Estructura-Actividad