bluuebunny/crossref_metadata_2025_split huggingface dataset:
Systemic bioavailability of hydrofluoroalkane (HFA) formulations of fluticasone/salmeterol in healthy volunteers via pMDI alone and spacer
Authors: Clearie Karine L. , Williamson Peter A. , Vaidyanathan Sriram , Du Bois Jeannine , Nell Haylene , Lipworth Brian J. 
Published: February 2010

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT • Seretide (GlaxoSmithKline, UK) is a combination inhaler which contains both fluticasone (FP) and salmeterol (SM).• A generic fluticasone/salmeterol (FP/SM) combination inhaler (Neolab, UK) has recently been developed.• Determination of therapeutic equivalence is essential in the development of generic products. WHAT THIS STUDY ADDS • This paper determines that in vitro fine particle dose may not predict pharmacokinetic and systemic pharmacodynamic outcomes.• This paper shows that the generic and reference products were equivalent in terms of FP, but not SM pharmacokinetics.• No significant differences were found between generic and reference products in terms of either adrenal suppression or hypokalaemia.AIM To compare a test version of HFA fluticasone/salmeterol (FP/SM) combination inhaler (Neolab, UK) with the reference product Seretide (GlaxoSmithKline, UK).METHODS An in vitro Anderson cascade impactor was used to compare the fine particle dose (<4.7 µm). Two separate randomized cross‐over studies were performed to compare the systemic bioavailability of test vs. reference (T vs. R) formulations of FP/SM 250/25 µg pMDI in healthy volunteers. In study 1 blood pharmacokinetic analysis using oral charcoal block was performed over 24 h following a single dose of four puffs via pMDI alone. In study 2 systemic bioactivity was measured following single doses of four and eight puffs via a spacer device: serum potassium (K+) to reflect SM, and overnight urinary cortisol : creatinine (OUCC) for FP. An early pharmacokinetic profile was also assessed over 120 min.RESULTS The in vitro fine particle dose was similar for test vs. reference pMDI alone and via spacer. The results of both studies were consistent: No significant differences between formulations were seen in terms of FP kinetics. Analysis of SM kinetics revealed superiority of the test product. No significant dose–response or difference in T : R ratio was noted for OUCC. Fall in K+ revealed a significant dose–response with a non‐significant T : R ratio.CONCLUSIONS The in vitro fine particle dose may not predict pharmacokinetic and systemic pharmacodynamic outcomes. Single dosing studies with fluticasone/salmeterol 250/25 µg via pMDI or with spacer showed pharmacokinetic equivalence with FP, but not SM. No significant difference between formulations was seen with either adrenal suppression or hypokalaemia.
Evaluation of Ki‐67 and Bcl‐2 antigen expression in breast carcinomas of women treated with raloxifene
Authors: Lopes‐Costa P. V. , Dos Santos A. R. , Dos Santos L. G. , Da Silva B. B. 
Published: February 2010

AbstractObjectives:  To evaluate the effect of raloxifene on Ki‐67 and Bcl‐2 antigen expression in operable, stage II, oestrogen‐receptor‐positive invasive ductal breast carcinomas.Materials and methods:  Twenty post‐menopausal women who had taken 60 mg of raloxifene daily for 28 days prior to definitive surgery were enrolled in the investigation. Two tumour samples were obtained by incisional biopsy during the study, one at the time of confirmation of diagnosis of invasive ductal carcinoma and evaluation of oestrogen receptor status, and the other 29 days later, at the time of definitive surgery. Immunohistochemistry was performed on tumour samples, prior to and after raloxifene treatment, to evaluate Ki‐67 and Bcl‐2 expression. Friedman and McNemar tests were used for statistical analysis of the data, significance being established at 5%.Results:  Mean percentage of Ki‐67‐stained nuclei was 24.86 ± 2.95 prior to raloxifene treatment and 13.33 ± 1.52 after treatment (P < 0.001). Prior to raloxifene treatment, only 9/20 cases (45%) were classified as Bcl‐2‐positive, whereas after treatment, 17/20 (85%) were classified as Bcl‐2‐positive (P < 0.013).Conclusions:  Raloxifene treatment significantly reduced Ki‐67 antigen expression and increased Bcl‐2 expression in breast carcinomas of post‐menopausal women.
Microplate spectrophotometry for high‐throughput screening of cytotoxic molecules
Authors: Tomelleri C. , Dalla Pellegrina C. , Chignola R. 
Published: February 2010

AbstractObjectives:  High‐throughput chemical and biochemical technologies are now exploited by modern pharmacology and toxicology to synthesize a multitude of new molecules with bioactive potential, or to isolate them from living matter. Testing molecules in cell systems on a large scale, however, is a rate‐limiting step in drug discovery or in toxicity assessment. In this study, we developed a low‐cost high‐throughput method for first‐level screening of cytotoxic molecules.Materials and methods:  We used microplate spectrophotometry to measure growth kinetics of human tumour cells that grow in suspension (Molt3) or adherent to the plastic surface of culture wells (HeLa) in standard RPMI medium. Cells were treated with colchicin, idarubicin or paclitaxel under various treatment schedules. The effects were quantified and compared with those measured by optical microscopy using the trypan blue dye exclusion method to reveal dead cells.Results:  Proliferation kinetics of tumour cells can be quantified by measuring variations in optical densities of cell samples at 410 and 560 nm wavelengths. For cells that grow in suspension, one single reading at 730 nm may be sufficient to reconstruct growth curves that parallel those obtained by direct cell counting. Effects of the cytotoxic treatments could also be quantified and results compared very favourably with those obtained using standard techniques.Conclusions:  Microplate spectrophotometry is a robust and sensitive method to monitor growth of animal cell populations both in the absence and in the presence of cytotoxic drugs. This method implements existing technologies and can be fully automated.
3‐O‐Methylfunicone, a metabolite produced by Penicillium pinophilum, modulates ERK1/2 activity, affecting cell motility of human mesothelioma cells
Authors: Buommino E. , Paoletti I. , De Filippis A. , Nicoletti R. , Ciavatta M. L. , Menegozzo S. , Menegozzo M. , Tufano M. A. 
Published: February 2010

AbstractObjectives:  3‐O‐methylfunicone (OMF), a secondary metabolite produced by Penicillium pinophilum, affects cell proliferation and motility in a variety of human solid tumours. The aim of this study was to demonstrate whether OMF has the ability to arrest cell division and motility, in a human mesothelioma cell line. Malignant mesothelioma is an aggressive cancer that does not respond to standard therapies the cells of which are considered to be highly resistant to apoptosis.Material and methods:  Cell motility and invasion were measured using a modified Boyden chamber. Gene expression was examined by RT‐PCR, while ERK1/2 was investigated by Western blot analysis. All experiments were also performed on primary cultures of mesothelial cells.Results:  The present study shows that OMF inhibited motility of the NCI mesothelioma cell line by modulating ERK signalling activity, and affected αVβ5 integrin and MMP‐2 expression, inducing marked downregulation at both mRNA and protein levels. Substantial downregulation of VEGF gene expression was also demonstrated. These effects were not observed in normal mesothelial cell cultures.Conclusion:  OMF may have potential as a naturally derived anti‐tumour drug for treatment of mesothelioma.
Analysis of radiation‐induced changes to human melanoma cultures using a mathematical model
Authors: Basse B. , Joseph W. R. , Marshall E. S. , Baguley B. C. 
Published: February 2010

AbstractObjectives:  Tumour cells respond to ionizing radiation by cycle arrest, cell death or repair and possible regrowth. We have developed a dynamic mathematical model of the cell cycle to incorporate transition probabilities for entry into DNA replication and mitosis. In this study, we used the model to analyse effects of radiation on cultures of five human melanoma cell lines.Materials and methods:  Cell lines were irradiated (9 Gy) prior to further culture and harvesting at multiple points up to 96 h later. Cells were fixed, stained with propidium iodide and analysed for G1‐, S‐ and G2/M‐phase cells by flow cytometry. Data for all time points were fitted to a mathematical model. To provide unique solutions, cultures were grown in the presence and absence of the mitotic poison paclitaxel, added to prevent cell division.Results:  The model demonstrated that irradiation at 9 Gy induced G2‐phase arrest in all lines for at least 96 h. Two cell lines with wild‐type p53 status additionally exhibited G1‐phase arrest with recovery over 15 h, as well as evidence of cell loss. Resumption of cycling of surviving cells, as indicated by increases in G1/S and G2/M‐phase transitions, was broadly comparable with results of clonogenic assays.Conclusions:  The results, combined with existing data from clonogenic survival assays, support the hypothesis that a dominant effect of radiation in these melanoma lines is the induction of long‐term cell cycle arrest.
Recombinant human midkine stimulates proliferation of articular chondrocytes
Authors: Zhang Z. H. , Li H. X. , Qi Y. P. , Du L. J. , Zhu S. Y. , Wu M. Y. , Lu H. L. , Yu Y. , Han W. 
Published: February 2010

AbstractObjectives:  Midkine, a heparin‐binding growth factor, promotes population growth, survival and migration of several cell types, but its effect on articular chondrocytes remains unknown. The aim of this study was to investigate its role on proliferation of articular chondrocytes in vitro and in vivo.Materials and methods:  Bromodeoxyuridine incorporation and MTT assays were performed to examine the proliferative effect of recombinant human midkine (rhMK) on primary articular chondrocytes. Activation of extracellular signal‐regulated kinase (ERK) and phosphatidylinositol 3‐kinase (PI3K) was analysed using western blot analysis. Systemic and local delivery of rhMK into mice and rats was preformed to investigate the proliferative effect of rhMK in vivo, respectively. Histological evaluation, including measurement of articular cartilage thickness, cell density, matrix staining and immunostaining of proliferating cell nuclear antigen was carried out.Results:  rhMK promoted proliferation of articular chondrocytes cultured in a monolayer, which was mediated by activation of ERK and PI3K. The proliferative role of rhMK was not coupled to dedifferentiation of culture‐expanded cells. Consistent with its action in vitro, rhMK stimulated proliferation of articular chondrocytes in vivo when it was administered subcutaneously and intra‐articularly in mice and rats, respectively.Conclusion:  Our results demonstrate that rhMK stimulates proliferation of primary articular chondrocytes in vitro and in vivo. The results of this study warrant further examination of rhMK for treatment of animal models of articular cartilage defects.
Patch size effects are more important than genetic diversity for plant–herbivore interactions in Brassica crops
Authors: HAMBÄCK PETER A. , BJÖRKMAN MARIA , HOPKINS RICHARD J. 
Published: February 2010

1. Considerable evidence suggests that the diversity within plant communities may strongly affect the strength of species interactions, but the majority of studies only considered interspecific diversity.2. This paper examines the effect of intraspecific genetic diversity within Brassica fields on two Brassica specialists, cabbage root fly, and diamondback moth, and on a parasitoid attacking diamondback moths. Genetic diversity was manipulated both in a replacement and an additive design.3. Both herbivore densities and parasitism rates were higher in smaller plots, with limited responses to increased within‐plot diversity. All species showed variable densities across genotypes, and preference hierarchies were species specific.4. Responses to plot size in root flies scaled with the diameter‐to‐area ratio, suggesting that patch detectability affected local density, whereas responses by diamondback moths and parasitoids deviated from this ratio. These species differences could be traced to differences in the residence time within patches, where diamondback moths typically spend longer and more variable time periods in patches than root flies.5. The lack of response to genetic diversity by both herbivores suggests that egg‐laying rates are affected by decisions on the plant and not by attraction from a distance, neither to the plant itself nor the patch. Patterns of differential attack may then be due to different acceptability for studied genotypes.6. Future theories on insect responses to spatial heterogeneity should focus on species traits and how traits interact with information landscapes in the field.
A novel frame‐shift deletion causing analbuminaemia in an Italian paediatric patient
Authors: Dagnino M. , Caridi G. , Marsciani M. , Bettocchi I. , Tassinari D. , Bernardi F. , Chiodo F. , Campagnoli M. , Galliano M. , Minchiotti L. 
Published: February 2010

Eur J Clin Invest 2010; 40 (3): 281–284AbstractBackground  Analbuminaemia (OMIM #103600) is a rare autosomal recessive disorder manifested by the absence or severe reduction of circulating serum albumin in homozygous or compound heterozygous subjects. The trait is caused by a variety of mutations within the albumin gene.Design  We report here the clinical and molecular characterization of a new case of congenital analbuminaemia in a 4‐year‐old Italian girl diagnosed on the basis of the low level of circulating albumin (= 10·0 g L−1). The albumin gene was screened by single‐strand conformation polymorphism and heteroduplex analysis and the mutated region submitted to DNA sequencing.Results  The proband was found to be homozygous, and both parents heterozygous, for a novel deletion in exon 8 (c.920delT). The subsequent frame‐shift should have given rise to a putative polypeptide chain of 304 amino acid residues, which we could not identify in the proband’s serum.Conclusions  A novel analbuminaemia causing mutation was identified and characterized at the clinical level in a child. The molecular diagnosis of the trait is based on the rapid localization of the mutation within the albumin gene by single‐strand conformation polymorphism and heteroduplex analysis, followed by DNA sequencing of the mutated region.
Cardiac hormones eliminate some human squamous lung carcinomas in athymic mice
Authors: Lenz A. , Sun Y. , Eichelbaum E. J. , Skelton W. P. , Pi G. , Vesely D. L. 
Published: February 2010

Eur J Clin Invest 2010; 40 (3): 242–249AbstractBackground  Four cardiac hormones synthesized by the same gene, i.e. atrial natriuretic peptide, vessel dilator, long acting natriuretic peptide and kaliuretic peptide, have anticancer effects in vitro.Materials and methods  These cardiac hormones were infused subcutaneously for 28 days with weekly fresh hormones at 0·3 nM kg−1 body weight in athymic mice bearing human squamous cell carcinomas.Results  Vessel dilator, atrial natriuretic peptide and kaliuretic peptide each eliminated one in six (17%) of the human squamous cell lung carcinomas. Long‐acting natriuretic peptide, although it did not eliminate any of the human squamous cell lung carcinomas did decrease the volume of one carcinoma to only 2% (P < 0·0001) of the untreated carcinomas. The squamous cell lung carcinomas that were not eliminated, with the exception of the one LANP‐treated tumour that decreased to only 2% of the volume of the untreated cancers, grew rapidly but their growth velocity compared to controls decreased by 76%, 40%, 38% and 25% in the vessel dilator, atrial natriuretic peptide, kaliuretic peptide and long‐acting natriuretic peptide groups respectively (P < 0·05).Conclusions  Three of four cardiac hormones synthesized by the atrial natriuretic peptide gene can eliminate human squamous cell lung carcinomas in athymic mice when treated subcutaneously for 4 weeks. The 4th cardiac hormone, i.e. long‐acting natriuretic peptide, decreased the volume of one squamous cell lung carcinoma to 2% of that of untreated animals, suggesting that it, too, has beneficial effects on squamous cell lung cancers.
Functional diversity is positively associated with biomass for lake diatoms
Authors: VOGT RICHARD J. , BEISNER BEATRIX E. , PRAIRIE YVES T. 
Published: February 2010

Summary1. Recent work has emphasised the benefit of using functional measures when relating biodiversity to ecosystem functioning. In this study, we investigated the extent to which functional and taxonomic diversity might be related to summed biovolume in community assemblages of 212 species of diatoms collected from 65 temperate lakes in western and central Quebec, Canada.2. We quantified functional diversity as both the total path‐length of a functional dendrogram (FD) and the variance in species traits (TV) for a given community. Selected traits included both size and responses to a set of environmental variables known to be influential for diatom communities.3. Species richness, as well as both FD and TV, was positively associated with total diatom biovolume at the level of the entire diatom community, suggesting that diversity in response types (particularly to total phosphorus and pH) is important for diatom community production.4. Although functional measures of diversity did not provide enhanced explanatory power over species richness, we argue that an exploration of functional traits potentially allows greater insight into the mechanisms underlying biodiversity–ecosystem functioning relations, indicating which traits might be most influential in driving community biomass production.
Regulation of nitrous oxide emission associated with benthic invertebrates
Authors: STIEF PETER , SCHRAMM ANDREAS 
Published: February 2010

Summary1. A number of freshwater invertebrate species emit N2O, a greenhouse gas that is produced in their gut by denitrifying bacteria (direct N2O emission). Additionally, benthic invertebrate species may contribute to N2O emission from sediments by stimulating denitrification because of their bioirrigation behaviour (indirect N2O emission).2. Two benthic invertebrate species were studied to determine (i) the dependence of direct N2O emission on the preferred diet of the animals, (ii) the regulation of direct N2O emission by seasonally changing factors, such as body size, temperature and availability and (iii) the quantitative relationship between direct and indirect N2O emission.3. Larvae of the mayfly Ephemera danica, which prefer a bacteria‐rich detritus diet, emitted N2O at rates of up to 90 pmol Ind.−1 h−1 under in situ conditions and 550 pmol Ind.−1 h−1 under laboratory conditions. In contrast, larvae of the alderfly Sialis lutaria, which prefer a bacteria‐poor carnivorous diet, emitted N2O at invariably low rates of 0–20 pmol Ind.−1 h−1. The N2O emission rate of E. danica larvae was positively correlated with seasonally changing factors (body size, temperature and availability). Direct N2O emission by E. danica larvae was limited by low temperature in winter, larval development in spring and low availability in summer.4. Both E. danica and the non‐emitting S. lutaria increased the total N2O and N2 emission from sediment in a density‐dependent manner. While N2O directly emitted by benthic invertebrates can be partially consumed in the sediment (E. danica), non‐emitting species can still indirectly contribute to total N2O emission from sediment (S. lutaria).
Differentiating aquatic plant communities in a eutrophic river using hyperspectral and multispectral remote sensing
Authors: TIAN YONG Q. , YU QIAN , ZIMMERMAN MARC J. , FLINT SUZANNE , WALDRON MARCUS C. 
Published: February 2010

Summary1. This study evaluates the efficacy of remote sensing technology to monitor species composition, areal extent and density of aquatic plants (macrophytes and filamentous algae) in impoundments where their presence may violate water‐quality standards.2. Multispectral satellite (IKONOS) images and more than 500 in situ hyperspectral samples were acquired to map aquatic plant distributions. By analyzing field measurements, we created a library of hyperspectral signatures for a variety of aquatic plant species, associations and densities. We also used three vegetation indices. Normalized Difference Vegetation Index (NDVI), near‐infrared (NIR)‐Green Angle Index (NGAI) and normalized water absorption depth (DH), at wavelengths 554, 680, 820 and 977 nm to differentiate among aquatic plant species composition, areal density and thickness in cases where hyperspectral analysis yielded potentially ambiguous interpretations.3. We compared the NDVI derived from IKONOS imagery with the in situ, hyperspectral‐derived NDVI. The IKONOS‐based images were also compared to data obtained through routine visual observations. Our results confirmed that aquatic species composition alters spectral signatures and affects the accuracy of remote sensing of aquatic plant density. The results also demonstrated that the NGAI has apparent advantages in estimating density over the NDVI and the DH.4. In the feature space of the three indices, 3D scatter plot analysis revealed that hyperspectral data can differentiate several aquatic plant associations. High‐resolution multispectral imagery provided useful information to distinguish among biophysical aquatic plant characteristics. Classification analysis indicated that using satellite imagery to assess Lemna coverage yielded an overall agreement of 79% with visual observations and >90% agreement for the densest aquatic plant coverages.5. Interpretation of biophysical parameters derived from high‐resolution satellite or airborne imagery should prove to be a valuable approach for assessing the effectiveness of management practices for controlling aquatic plant growth in inland waters, as well as for routine monitoring of aquatic plants in lakes and suitable lentic environments.
Land use and season affect fluxes of CO2, CH4, CO, N2O, H2 and isotopic source signatures in Panama: evidence from nocturnal boundary layer profiles
Authors: PENDALL E. , SCHWENDENMANN L. , RAHN T. , MILLER J. B. , TANS P. P. , WHITE J. W. C. 
Published: February 2010

AbstractConversion of tropical rainforests to pastures and plantations is associated with changes in soil properties and biogeochemical cycling, with implications for carbon cycling and trace gas fluxes. The stable isotopic composition of ecosystem respiration (δ13CR and δ18OR) is used in inversion models to quantify regional patterns of CO2 sources and sinks, but models are limited by sparse measurements in tropical regions. We measured soil respiration rates, concentrations of CO2, CH4, CO, N2O and H2 and the isotopic composition of CO2, CH4 and H2 at four heights in the nocturnal boundary layer (NBL) above three common land‐use types in central Panama, during dry and rainy seasons. Soil respiration rates were lowest in Plantation (average 3.4 μmol m−2 s−1), highest in Pasture (8.3 μmol m−2 s−1) and intermediate in Rainforest (5.2 μmol m−2 s−1). δ13CR closely reflected land use and increased during the dry season where C3 vegetation was present. δ18OR did not differ by land use but was lower during the rainy than the dry season. CO2 was correlated with other species in approximately half of the NBL profiles, allowing us to estimate trace gas fluxes that were generally within the range of literature values. The Rainforest soil was a sink for CH4 but emissions were observed in Pasture and Plantation, especially during the wet season. N2O emissions were higher in Pasture and Plantation than Rainforest, contrary to expectations. Soil H2 uptake was highest in Rainforest and was not observable in Pasture and Plantation during the wet season. We observed soil CO uptake during the dry season and emissions during the wet season across land‐use types. This study demonstrated that strong impacts of land‐use change on soil–atmosphere trace gas exchange can be detected in the NBL, and provides useful observational constraints for top‐down and bottom‐up biogeochemistry models.