38,845 research outputs found
Simultaneous, phenotypic knockout of VEGF-R2 and Tie-2 with an intradiabody enhances antiangiogenic effects in vivo
No full textBackground: Intracellular antibodies (intrabodies) have been used for the generation of phenotypic knockouts in vivo by surface depletion of extracellular or transmembrane proteins. Intrabodies present an alternative to methods of gene inactivation that target genomic DNA or m-RNA, such as RNA interference. Several studies suggest that the VEGF receptor pathway and the Tie-2 pathway are independent and essential mediators of angiogenesis, leading to the hypothesis that simultaneous interference with both pathways should result in additive effects in tumor growth. Methods: In order to generate a precise tool for the simultaneous silencing of two independent signaling pathways essential for angiogenesis, we developed a bispecific, tetravalent endoplasmatic reticulum (ER)-targeted intradiabody, against Tie-2 and VEGF-R2. Results: Using an adenovirus mediated gene delivery system, we achieved the simultaneous downregulation of the two cell surface receptors and demonstrate that the intradiabody is significantly more powerful with respect to efficiency and duration of surface depletion of Tie-2 and VEGF-R2 when compared to scFv intrabodies. In a human melanoma xenograft mouse model, we could show that blockade of both VEGF-R2 and Tie-2 pathways or the VEGF receptor pathway alone resulted in a significant inhibition of tumor growth and tumor angiogenesis (92.2% and 74.4%). Conclusion: We demonstrate for the first time that simultaneous inhibition of the VEGF and the Tie-2 receptor pathways result in additive antiangiogenic effects in vitro and in vivo as compared to single VEGF receptor pathway blockade, strengthening the potential of simultaneous targeting of multiple pathways as a therapeutic strategy
Erratum to: Effect of moderate red wine intake on cardiac prognosis after recent acute myocardial infarction of subjects with Type 2 diabetes mellitus (Diabetic Medicine, (2006), 23, 9, (974-981), 10.1111/j.1464-5491.2006.01886.x)
No full textIn an article by Marfella et al, the author name C. Saron is incorrect and should be listed as C. Sardu. Therefore the correct author list is: R. Marfella, F. Cacciapuoti, M. Siniscalchi, F. C. Sasso, F. Marchese, F. Cinone, E. Musacchio, M. A. Marfella, L. Ruggiero, G. Chiorazzo, D. Liberti, G. Chiorazzo, G. F. Nicoletti, C. Sardu, F. D'Andrea, C. Ammendola, M. Verza and L. Coppola.In an article by Marfella et al, the author name C. Saron is incorrect and should be listed as C. Sardu. Therefore the correct author list is: R. Marfella, F. Cacciapuoti, M. Siniscalchi, F. C. Sasso, F. Marchese, F. Cinone, E. Musacchio, M. A. Marfella, L. Ruggiero, G. Chiorazzo, D. Liberti, G. Chiorazzo, G. F. Nicoletti, C. Sardu, F. D'Andrea, C. Ammendola, M. Verza and L. Coppola
Cover Picture: Towards Organo‐Click Chemistry: Development of Organocatalytic Multicomponent Reactions Through Combinations of Aldol, Wittig, Knoevenagel, Michael, Diels–Alder and Huisgen Cycloaddition Reactions (Chem. Eur. J. 21/2004)
No full textThe cover picture shows a perspective of organo-click chemistry depicting the bringing together of simple reagents, and amino acid and copper/copper sulfate catalysts to produce spirotriones, which are antioxidant compounds. Unlike conventional methods, organo-click chemistry produces stereospecific products with very good yields in a single operation. Multicomponent reactions catalyzed by amino acids and copper sulfate together with simple reagents depicted here support the potential of a rich variety of chemistries available to the prebiotic world and beyond. For more information see the article by D. B. Ramachary and C. F. Barbas III on p. 5323 ff. The picture was designed by Michael Pique at the Scripps Research Institute
Development of chromatographic methods for the determination of genotoxic impurities in cloperastine fendizoate
No full textThe classification of an impurity of a drug substance as genotoxic means that the “threshold of toxicological concern” (TTC) value of 1.5 ug/day intake, considered to be associated with an acceptable risk, should be the admissible limit in the raw material and that leads to new analytical challenges. In this study, reliable chromatographic methods were developed and applied as limit tests for the control of three genotoxic impurities (GTIs) in cloperastine fendizoate, drug widely used as an antitussive active pharmaceutical ingredient (API). In particular, GC–MS was applied to the determination of one alkyl halide (2-chloroethanol, 2-CE), while HPLC-DAD was selected for the analysis of two sulfonate esters (methyl p-toluenesulfonate, MPTS, and 2-chloroethyl p-toluenesulfonate, CEPTS).Regarding GC–MS, strong anion-exchange (SAX)-SPE was applied to remove fendizoate from the sample solutions, due its low volatility and its high amount in the raw material. The GC–MS analysis was performed on a Factor Four VF-23ms capillary column (30 m × 0.25 mm I.D., film thickness 0.25 um, Varian). Single ion-monitoring (SIM) detection mode was set at m/z 80.In the case of HPLC-DAD, a suitable optimization of the chromatographic conditions was carried out in order to obtain a good separation of the impurity peaks from the drug substance peaks. The optimized method utilizes a SymmetryShield RP8 column (250 mm × 4.6 mm, 5 um, Waters) kept at 50 ◦C, withphosphate buffer (pH 3.0; 10 mM)–methanol (containing 10% ACN) (45:55, v/v) as the mobile phase, at the flow-rate of 1.7 mL/min and UV detection at 227 nm. The required sensitivity level was achieved by injecting 80 uL of sample solution, purified from fendizoate by SAX-SPE, followed by a 1:1 (v/v) dilutionof the SPE eluate with water.For both GC–MS and HPLC-DAD, the method validation was performed in relation to specificity and limit of detection (LOD), as required by ICH guidelines in relation to limit assays. The developed methods were successfully applied for the determination of GTIs in five different batches of cloperastine fendizoate. In all the analyzed batches, the three target GTIs were below the concentration limit
METABOLOMIC ASSESSMENT WITH LC-QTOF-MS OF THE NUTRACEUTICAL EFFECT OF ROSMARINUS OFFICINALIS EXTRACTS ON PLASMA SAMPLES OF DIABETIC RATS
No full textDiabetes mellitus is one of the major worldwide health problems. The peculiarity of this disease is to have high levels of oxidative stress, which plays a crucial role in the pathogenesis of other complications, such as nephropathy, neuropathy and retinopathy. Rosmarinus officinalis L. extracts, processed by supercritical fluid extraction (SFE), have long been recognized as having antioxidant properties. To investigate the potential nutraceutical properties of these extracts on type 1 diabetes, a metabolomic fingerprinting study was carried out using LC-QTOF-MS. Metabolomic fingerprinting is a technique which study as many analytes as possible to get a complete view of a disease and of the different levels of metabolites involved, and finally to identify possible biomarkers.In this work, Sprague-Dawley male rats (12 ± 2 weeks of age), treated with streptozotocin, were used in controlled conditions in which they received, by intragastric administration, five doses of rosemary extract containing 10% folic acid (used to improve the endothelial progenitor cell function) dispersed in 1 mL vehicle or only 1 mL of vehicle. There were also healthy rats that received the same treatment, thus generating four groups of samples: treated or non-treated (only vehicle) controls, treated and non-treated diabetics.Plasma fingerprints of control and diabetic rats, focused on the most polar constituents, were obtained by LC-QTOF-MS. Retention of polar compounds from plasma in hydrophilic interaction liquid chromatography (HILIC) mode and use of solvents readily compatible with mass spectrometry is a new approach that was initially explored by testing different stationary phases. The HILIC columns used in this study included an Ascentis Si (15 cm × 2.1 mm, 5 μm, Supelco), a TSK gel Amide-80 (5 cm × 2.0 mm, 3 μm, Tosoh), a Luna HILIC (10 cm × 2.0 mm, 5 μm, Phenomenex) and a ZIC-HILIC (10 cm × 2.1 mm, 3.5 μm, Merck). Unfortunately, the results were not as good as it was expected and this was mainly due to the lack of precision. It was therefore decided to analyze the plasma samples under reversed-phase (RP) conditions, using a Discovery HS C18 column (15 cm × 2.1 mm, 3 μm, Supelco), with a mobile phase composed by water (with 0.1% of formic acid) and ACN (with 0.1% of formic acid), under gradient elution. The injection volume was 10 μL; the flow rate was 0.6 mL/min and the column temperature was set at 30 °C. Data were collected in the positive ion mode with a QTOF mass analyzer, operated in the full scan mode from 50 to 1000 m/z. Experimental data from RP-HPLC were further processed by multivariate analysis to find compounds with statistically significant changes. Then, MS/MS experiments were performed under the same experimental conditions and identifications were carried out by studying the fragmentation pattern of the target analytes or using reference standards. Finally, the biochemical interpretation of potential biomarkers associated with the disease was performed. The compounds that were found to be affected by the nutraceutical treatment include:•Lysophosphatidylcholines (LPCs), represent the first group of identified compounds. LPCs are generated from ox-LDL or from inflammatory cells, and are very important intermediates of different metabolic pathways. These compounds were found to increase in diabetic rats after the treatment.•Pipecolic acid, the major metabolite of lysine degradation. This compound was found to increase in diabetic rats after the treatment.•Lipoic acid, an antioxidant synthesized in mitochondria. It is a necessary co-factor for mitochondrial α-ketoacid dehydrogenases, and thus plays a critical role in mitochondrial energy metabolism. This metabolite was found to increase in diabetic rats after the treatment.•Carnitine and derivates, which is important to transport fatty acids into the mitochondria. Also these compounds were found to be affected by the treatment.In conclusion, the LC-QTOF-MS analysis allowed us to study a large number of compounds, very useful in metabolomic fingerprinting, and, with the help of chemometric analysis, to select masses with statistically significant changes. The levels of LPCs, pipecolic acid, lipoic acid, carnitine and derivates were found to be improved after the nutraceutical treatment, that can be considered useful to counteract some of the deleterious effects of type 1 diabetes
Reptiles en corredores biológicos y mortalidad por atropellamiento vehicular en Barbas-Bremen, departamento del Quindío, Colombia
No full textLos corredores biológicos de Barbas-Bremen son una estrategia de conservación implementada en el departamento del Quindío, Colombia, con el objetivo de reestablecer la conectividad entre poblaciones bióticas presentes en los dos fragmentos de bosque más grandes de la región. No obstante, pocos estudios han evaluado la efectividad de estos corredores como hábitat para especies de fauna local y como facilitadores de dispersión de individuos entre las reservas naturales de Barbas y Bremen. Caracterizamos la fauna de reptiles en tres corredores biológicos y se evaluó si la mortalidad de reptiles por atropellamiento vehicular en una carretera que atraviesa dichos corredores biológicos está correlacionada con la distancia a ellos. Utilizamos encuentros visuales libres y trampas de caída para muestrear la riqueza de reptiles en cada corredor biológico y áreas de pastizal adyacentes; los individuos atropellados los registramos durante 34 caminatas en la carretera. Encontramos 42 individuos pertenecientes a 16 especies; los reptiles en los corredores biológicos tienden a ser arbóreos mientras que los de pastizales tienden a ser terrestres. No hubo relación entre el número de atropellos (63 serpientes, 9 especies) y la distancia al corredor biológico más cercano. En general, los corredores biológicos de Barbas-Bremen ofrecen un hábitat boscoso a numerosas especies de reptiles que probablemente no podrían subsistir en la matriz de pastizales que predomina en el área de estudio; sin embargo, más estudios son necesarios para establecer si dichos corredores biológicos favorecen el flujo de individuos entre Barbas y Bremen y como la carretera ahí presente afectaría este proceso.The biological corridors of Barbas-Bremen are a conservation strategy implemented in the department of Quindío, Colombia, in order to reestablish connectivity between biotic populations in the two largest forest fragments in the region. However, very few studies have evaluated the effectiveness of those corridors as habitat for local wildlife and as facilitators of dispersal of individuals between the natural reserves of Barbas and Bremen. Between July 2014 and March 2015 we performed 18 fieldtrips to characterize the reptile fauna in three biological corridors and test whether mortality of reptiles on a road that crosses the biological corridors is correlated with distance from them. We used visual sampling and pitfall traps to sample the richness of reptiles in each biological corridor and adjacent grassland; roadkills were recorded through 34 walks on the road. We found 42 reptiles belonging to 16 species; reptiles in biological corridors tend to be arboreal, the species in grassland tend to be terrestrial. There was no relationship between the number of roadkills (64 snakes, 9 species) and the distance to the nearest biological corridor. We do not observe lizards killed on the road. In general, biological corridors of Barbas-Bremen offer a forested habitat to numerous species of reptiles that probably could not survive in the grassland that prevails in the study area; however, more research is required to determine whether these biological corridors encourage the flow of individuals between Barbas and Bremen, and the role that the road there plays in this process
Mitomycin C in highly myopic eyes - Author reply
No full textOphthalmology. 2005 Feb;112(2):208-18; discussion 219.
Mitomycin C modulation of corneal wound healing after photorefractive keratectomy in highly myopic eyes.
Gambato C, Ghirlando A, Moretto E, Busato F, Midena E.
SourceRefractive Surgery Service and Antimetabolite Therapy Research Unit, Department of Ophthalmology, University of Padova, Padova, Italy.
Abstract
PURPOSE: To evaluate the role of topical mitomycin C in corneal wound healing (CWH) after photorefractive keratectomy (PRK) in highly myopic eyes.
DESIGN: Prospective, double-masked, randomized clinical trial.
PARTICIPANTS: Seventy-two eyes of 36 patients affected by high (>7 diopters) myopia.
METHODS: In each patient, one eye was randomly assigned to PRK with intraoperative topical 0.02% mitomycin C application, and the fellow eye was treated with a placebo. Postoperatively, mitomycin C-treated eyes received artificial tears (3 times daily, tapered in 3 months), whereas the fellow eye was treated with fluorometholone sodium 2% and artificial tears (3 times daily, tapered in 3 months).
MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA) and best-corrected visual acuity (BCVA), contrast sensitivity, manifest refraction, and biomicroscopy. Contrast sensitivity was determined using the Pelli-Robson chart. Corneal confocal microscopy documented CWH.
RESULTS: Mean follow-up was 18 months (range, 12-36). No side effects or toxic effects were documented. At 12-month follow-up examination, UCVAs (logarithm of the minimum angle of resolution) were 0.4+/-0.48 and 0.5+/-0.53 (P = .03) in mitomycin C-treated eyes and corticosteroid-treated eyes, respectively. At 1 year, corneal haze developed in 20% of corticosteroid-treated eyes, versus 0% of mitomycin C-treated eyes. At 12, 24, and 36 months, corneal confocal microscopy showed activated keratocytes and extracellular matrix significantly more evident in untreated eyes (Ps = 0.004, 0.024, and 0.046, respectively).
CONCLUSION: Topical intraoperative application of 0.02% mitomycin C can reduce haze formation in highly myopic eyes undergoing PRK.
Comment in
Ophthalmology. 2006 Feb;113(2):357; author reply 357-8
Bromelias epífitas de los corredores biológicos del distrito de conservación de suelos barbas Bremen en el municipio de Finlandia Quindío
No full textTablas, figurasLas bromelias epífitas son un componente importante de los ecosistemas terrestres debido a su alta diversidad en particular en la región Andina de Colombia, su contribución a los ciclos de nutrientes, proporción de hábitats para organismos terrestres y acuáticos. Los estudios sobre su diversidad y las posibles causas de perdida de esta son relevantes para formular estrategias de conservación bajo los escenarios actuales de deforestación y urbanización en la región Andina. El objetivo de este estudio fue conocer la diversidad de bromelias en un área protegida de los Andes Colombianos, así como su perdida por factores que causan la caída de bromelias dentro del área protegida. El primer capítulo describe las bromelias epífitas en cuanto a su riqueza, abundancia y composición taxonómica, comparando la comunidad de bromelias entre los cuatro corredores presentes en el área protegida. En este primer estudio se encontraron 10 especies de bromelias epífitas distribuidas en 5 géneros (Guzmania, Mezobromelia, Racinea, Tillandsia y Vriesea), dentro de las cuales Tillandsia complanata presentó la mayor abundancia. La abundancia y composición de especies difirió entre los corredores producto de las diferencias en las características de los árboles hospederos (forofitos). Además, este capítulo contribuye a la ampliación de la distribución geográfica para la especie G. cundinamarcae, la cual solo se había reportado para Cundinamarca, Colombia. El segundo capítulo analizó y evaluó cómo factores ambientales (precipitación), mecánicos (el tamaño de las ramas de los árboles) y rasgos de las bromelias influyen en la caída de bromelias tanque y por ende en la perdida de especies para el ecosistema. Las especies G. cundinamarcae y T. complanata fueron las que con mayor abundancia se encontraron caídas. Los aumentos en las precipitaciones incrementaron el número de bromelias caídas principalmente de tamaño pequeño, etapa juvenil y buen estado fitosanitario. Estos resultados son relevantes desde la perspectiva de los futuros escenarios de cambio climático (incrementos en las precipitaciones) para la región de los Andes. Además, nuestros resultados subrayan la importancia de implementar estrategias de conservación de bromelias como la recuperación de las bromelias caídas, las cuales al no ser rescatadas serían consideradas muertas incrementando los valores de perdida de diversidad.Epiphytic bromeliads are an important component of terrestrial ecosystems due to their high diversity, particularly in the Andean region of Colombia, their contribution to nutrient cycles, habitat proportion for terrestrial and aquatic organisms. Studies o n its diversity and the possible causes of its loss are relevant to formulate conservation strategies under the current scenarios of deforestation and urbanization in the Andean region. The objective of this study was to know the diversity of bromeliads in a protected area of the Colombian Andes, as well as their loss due to factors that cause the fall of bromeliads within the protected area. The first chapter describes the epiphytic bromeliads in terms of their richness, abundance, and taxonomic composition, comparing the bromeliad community between the four corridors present in the protected area. This first study found 10 species of epiphytic bromeliads distributed in 5 genera ( Guzmania, Mezobromelia, Racinea, Tillandsia and Vriesea ), among which Tillands ia complanata presented
the highest abundance. The abundance and species composition differed between corridors due to differences in the characteristics of the host trees . In addition, this chapter contributes to the extension of the geographic distribution for the species G. cundinamarcae , which had only been reported from Cundinamarca, Colombia. The second chapter analyzed and evaluated how environmental (precipitation), mechanical (the size of tree branches) and bromeliad traits influence the fall of tank bromeliads and thus the loss of species to the ecosystem. The species G. cundinamarcae and T. complanata were the most abundant species found to have fallen. Increases in rainfall increased the number of fallen bromeliads, mainly of small size, juvenile stage and good phytosanitary condition. These results are relevant from the perspective of future climate change scenarios (increases in precipitation) for the Andes region. In addition, our results underline the importance of implementing bromeliad conservation str ategies such as the recovery of fallen bromeliads, which if not rescued would be considered dead, increasing the values of loss of diversity.Bromelias epífitas de los corredores biológicos del Distrito de Conservación de Suelos Barbas Bremen en el municipio de Filandia Quindío -- Agradecimientos -- Dedicatoria -- Declaración de Originalidad -- Resumen -- Abstract -- Introducción -- Referencias -- Capítulo 1: Host tree height drives the epiphytic bromeliad diversity in biological corridors of the Barbas Bremen Soil Conservation District -- Abstract -- Resumen -- Introduction -- Methods -- Results -- Discussion -- Referencias -- Anexo 1 -- Capítulo 2: Drivers of Bromeliad Fall and Implications for Epiphyte Conservation in Andean Tropical Forests -- Abstract -- Introduction -- Material and Methods -- Results -- Discussion -- Conclusiones -- Anexo 2Listado de tablas -- Capítulo 1: Table 1. Abundance and species richness of epiphytic bromeliads in the biological corridors of the Barbas Bremen Soil Conservation District -- Table S1. Distances between the biological corridors of Barbas Bremen -- Table S2. Relative distribution of epiphytic bromeliads: average and standard deviation by species, corridor, and transect in the biological corridors of the Barbas Bremen Soil Conservation District -- Capítulo 2: Table 1. Relation between canopy bromeliads and fallen bromeliads at the population level (LME = linear mixed model, x² = chi-square, df = degrees of freedom, p = probability; data with the symbol * represent a significant effect) -- Table 2. Effect of corridor and precipitation on the fallen bromeliad in the community and population level (GAMM = generalized additive model, df = degrees of freedom, F = Fisher test, p = probability; * = significant effect) -- Table 3. Effect of corridor and precipitation on the morphological characteristics, developmental stage, and phytosanitary status of 156 individuals of fallen bromeliads in the biological corridors of the Barbas Bremen Soil Conservation District (GAMM = generalized additive model, df = degrees of freedom, F = Fisher test, p = probability; * = significant effect) -- Table S1. Number of individuals of canopy and fallen bromeliad species in the biological corridors of the Barbas Bremen Soil Conservation District -- Listado de figuras -- Capítulo 1: Figure 1. Map of the biological corridors of the Barbas Bremen Soil Conservation District (Filandia-Quindío) and sampling points -- Figure 2. Abundance of epiphytic bromeliad species in the biological corridors of the Barbas Bremen Soil Conservation District -- Figure 3. DBH and height of host trees of epiphytic bromeliads in the biological corridors of the Barbas Bremen Soil Conservation District -- Figure 4. Range-abundance curve of epiphytic bromeliads in the biological corridors of the Barbas-Bremen Soil Conservation District (G.c = Guzmania cundinamarcae, G.m = Guzmania multiflora, G.sp1 = Guzmania sp1, M.c = Mezobromelia capituligera, R.sp1 = Racinea sp1, T.b = Tillandsia biflora, T.c = Tillandsia complanata, T.f = Tillandsia fendleri, T.sp1 = Tillandsia sp1, V.t = Vriesea tequendamae) -- Figure S1. Heat map showing the composition of transects and epiphytic bromeliads in the biological corridors (M = Monos aulladores, L = Laureles, P = Pavas, C = Colibríes) -- Capítulo 2: Fig. 1. Map of the biological corridors of the Barbas Bremen Soil Conservation District (Filandia-Quindío) and sampling points -- Fig. 2. Abundance of fallen bromeliads by species in the biological corridors of the Barbas Bremen Soil Conservation District -- Fig. 3. Relationship between the canopy bromeliad community and the fallen bromeliad community (A. Richness of fallen bromeliad species; B. Abundance of fallen bromeliads; gray lines represent the 95% confidence interval) -- Fig. 4. Effects of precipitation and corridors on richness (A), abundance (B), morphological characteristics (C), phytosanitary status (D), and developmental status (E) of fallen bromeliads in the Barbas Bremen Soil Conservation District -- Fig. 5. GAMM model for DBH of fallen branches with respect to the number of fallen bromeliads per branch in the biological corridors of the Barbas Bremen Soil Conservation District -- Fig. S1. Relationship of fallen bromeliad population size with canopy-bromeliad population size in the biological corridors of the Barbas Bremen Soil Conservation District -- Fig. S2. Frequency histograms of the morphological characteristics of fallen bromeliads and mechanical factor (diameter of fallen branch) in the biological corridors of the Barbas Bremen Soil Conservation District -- Fig. S3. Precipitation behavior every 15 days in the biological corridors of the Barbas Bremen Soil Conservation District.MaestríaMagister en Ciencias Biológica
Six Overtures Composed by C. F. Abel. Adapted for the Harpsichord or Piano Forte : being Opera First / By the Author
No full textSIX OVERTURES COMPOSED BY C. F. ABEL. ADAPTED FOR THE HARPSICHORD OR PIANO FORTE : BEING OPERA FIRST / BY THE AUTHOR
Six Overtures Composed by C. F. Abel. Adapted for the Harpsichord or Piano Forte : being Opera First / By the Author (1)
Cover (1)
Titelseite (2)
Overture I. (3)
Overture II. (8)
Overture III. (12)
Overture IV. (16)
Overture V. (20)
Overture VI. (24
- …
